title stringlengths 0 1.13k | abstract stringlengths 1 15.7k | PMID int64 22 36.5M |
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Videotaped experiments to drop safety behaviors and self-focused attention for patients with social anxiety disorder: do they change subjective and objective evaluations of anxiety and performance? | Safety behavior (SB) and self-focused attention (SFA) have been posited as important maintenance factors in the cognitive model of social anxiety disorder (SAD). The present study reports the results of experiments to drop SB and SFA among clinically diagnosed patients with SAD employing their own idiosyncratic anxiety-provoking situations. The ratings for observable anxiety, belief in feared outcome and overall performance were better for role plays without SB and SFA than for role plays with them. The degree of drop in SFA predicted drop in observable anxiety and belief in feared outcome. Dropping SB and SFA, however, was unable to completely correct the cognitive distortion because the subjective ratings were still significantly worse than the objective ratings. | 18,930,452 |
Investigation of folic acid stability in fortified instant noodles by use of capillary electrophoresis and reversed-phase high performance liquid chromatography. | A single enzyme treatment with alpha-amylase, prior to the quantification of added folic acid (FA) in fortified instant fried Asian noodles with analysis performed by capillary zone electrophoresis (CZE) and reversed-phase high performance liquid chromatography (RP-HPLC) with UV detection, is described. The method was validated and optimized for capillary electrophoresis (CE) with separation achieved using a 8 mM phosphate-12 mM borate run buffer with 5% MeOH at pH 9.5. FA was well separated from matrix components with nicotinic acid (NA) employed as an internal standard. In a comparative study, separation of FA was performed using HPLC with a mobile phase consisting of 27% MeOH (v/v) in aqueous potassium phosphate buffer (3.5 mM KH(2)PO(4) and 3.2 mM K(2)HPO(4)), pH 8.5, and containing 5 mM tetrabutylammonium dihydrogen phosphate as an ion-pairing agent. For both methods, excellent results were obtained for various analytical parameters including linearity, accuracy and precision. The limit of detection was calculated to be 2.2 mg/L for CE without sample stacking and 0.10 mg/L with high performance liquid chromatography (HPLC). Sample extraction involved homogenization and enzymatic extraction with alpha-amylase. Results indicated that FA was stable during four main stages of instant fried noodle manufacturing (dough crumbs, cut sheets, steaming and frying). | 18,930,464 |
The role of membranous urethral afferent autonomic innervation in the continence mechanism after nerve sparing radical prostatectomy: a clinical and prospective study. | We evaluated the somatic and autonomic innervation of the pelvic floor and rhabdosphincter before and after nerve sparing radical retropubic prostatectomy using neurophysiological tests and correlated findings with clinical parameters and urinary continence. From February 2003 to October 2005, 46 patients with prostate cancer were enrolled in a controlled, prospective study. Patients were evaluated before and 6 months after nerve sparing radical retropubic prostatectomy using the UCLA-PCI urinary function domain and neurophysiological tests, including somatosensory evoked potential, and the pudendo-urethral, pudendo-anal and urethro-anal reflexes. Clinical parameters and urinary continence were correlated with afferent and efferent innervation of the membranous urethra and pelvic floor. We used strict criteria to define urinary continence as complete dryness with no leakage at all, not requiring any pads or diapers and with a UCLA-PCI score of 500. Patients with a sporadic drop of leakage, requiring up to 1 pad daily, were defined as having occasional urinary leakage. Two patients were excluded from study due to urethral stricture postoperatively. We evaluated 44 patients within 6 months after surgery. The pudendo-anal and pudendo-urethral reflexes were unchanged postoperatively (p = 0.93 and 0.09, respectively), demonstrating that afferent and efferent pudendal innervation to this pelvic region was not affected by the surgery. Autonomic afferent denervation of the membranous urethral mucosa was found in 34 patients (77.3%), as demonstrated by a postoperative increase in the urethro-anal reflex sensory threshold and urethro-anal reflex latency (p <0.001 and 0.0007, respectively). Six of the 44 patients used pads. One patient with more severe leakage required 3 pads daily and 23 showed urinary leakage, including 5 who needed 1 pad per day and 18 who did not wear pads. Afferent autonomic denervation at the membranous urethral mucosa was found in 91.7% of patients with urinary leakage. Of 10 patients with preserved urethro-anal reflex latency 80% were continent. Sensory and motor pudendal innervation to this specific pelvic region did not change after nerve sparing radical retropubic prostatectomy. Significant autonomic afferent denervation of the membranous urethral mucosa was present in most patients postoperatively. Impaired membranous urethral sensitivity seemed to be associated with urinary incontinence, particularly in patients with occasional urinary leakage. Damage to the afferent autonomic innervation may have a role in the continence mechanism after nerve sparing radical retropubic prostatectomy. | 18,930,493 |
Pretreatment predictors of death from other causes in men with prostate cancer. | Most men diagnosed with prostate cancer will die of other causes and pretreatment patient characteristics may identify those who are likely to die of other causes. Accurate stratification of patients by risk of other cause mortality may reduce needless treatment preventing morbidity and expense. Using the CaPSURE database a cohort of men was identified with clinically localized prostate cancer who had definitive treatment with radical prostatectomy or radiation therapy between 1995 and 2004. Pretreatment patient characteristics were evaluated to determine if early other cause mortality could be predicted. Of 13,124 subjects enrolled in CaPSURE 5,070 had clinical T1c-T3a prostatic adenocarcinoma treated with radical prostatectomy (77%) or radiation therapy (23%) and posttreatment followup data. Median followup was 3.3 years. The cohort was divided into 3 groups. The prostate cancer specific mortality group included 55 men (1%) who died of prostate cancer. The 296 men (6%) who died of causes other than prostate cancer comprised the other cause mortality group. A third group contained the 4,719 (93%) men surviving at the end of the observation period. Factors that exclusively predicted death from nonprostate cancer causes included age at diagnosis, having a high school education or less, high clinical risk, smoking at time of diagnosis, concurrent nonprostate malignancy and worse scores on the Short Form-36 Health Survey physical function scale. Several pretreatment patient characteristics may identify patients at high risk of nonprostate cancer mortality. Future studies should consider stratifying patients by or at least reporting these variables. | 18,930,498 |
High anterior release of the levator fascia improves sexual function following open radical retropubic prostatectomy. | Recent anatomical studies have shown that branches of the cavernous nerves running adjacent to the prostate at the apex travel more anteriorly than previously recognized. Outcomes of robot assisted radical prostatectomy suggest improved postoperative sexual outcomes following high anterior release of the levator fascia. We prospectively evaluated the effect of high anterior release on oncological and sexual function outcomes following open radical retropubic prostatectomy. A total of 167 patients with clinically localized prostate cancer with a preoperative Sexual Health Inventory for Men score of greater than 21 underwent radical retropubic prostatectomy with bilateral nerve sparing and selective high anterior release, as performed by a single surgeon. Data on postoperative sexual function were collected by an independent third party. Sexual function outcomes at 12 months were defined as 1) a Sexual Health Inventory for Men score of 16 or greater and/or a satisfaction score of 4 or greater and 2) a Sexual Health Inventory for Men score of 22 or greater. Because unilateral high anterior release was equivalent to bilateral high anterior release for both definitions (p >0.3), they were combined into 1 group for analyses. Patients undergoing high anterior release were more likely to achieve a Sexual Health Inventory for Men score of 16 or greater and/or a satisfaction score of 4 (93% vs 77%, p = 0.007), and a Sexual Health Inventory for Men score of 22 or greater (70% vs 54%, p = 0.07) at 1 year. Return to baseline (a Sexual Health Inventory for Men score of 22 or greater) was even higher among patients receiving high anterior release who were more sexually active (greater than 1 attempt per week) preoperatively (78% vs 52%, p <0.05). The improved outcomes in potency achieved with high anterior release did not increase the likelihood of a positive surgical margin. Unilateral or bilateral high anterior release of the levator fascia in open radical retropubic prostatectomy provides excellent oncological results and is associated with improved postoperative sexual function. | 18,930,504 |
Glycan deletions in the HIV-1 gp120 V1/V2 domain compromise viral infectivity, sensitize the mutant virus strains to carbohydrate-binding agents and represent a specific target for therapeutic intervention. | Carbohydrate-binding agents (CBAs), such as the mannose-specific Hippeastrum hybrid agglutinin (HHA) and the GlcNAc-specific Urtica dioica agglutinin (UDA), frequently select for glycan deletions in all different domains of HIV-1 gp120, except in the V1/V2 domain. To reveal the underlying mechanisms, a broad variety of 31 different virus strains containing one or several N-glycan deletions in V1/V2 of the gp120 of the X4-tropic HIV-1(NL4.3) were constructed by chimeric virus technology. No co-receptor switch to CCR5 was observed for any of the replication-competent mutant virus strains. With a few exceptions, the more glycans were deleted in the gp120 V1/V2 domain, the more the replication capacity of the mutant viruses became compromised. None of the mutant virus strains showed a markedly decreased sensitivity to the inhibitory activity of HHA and UDA. Instead, an up to 2- to 10-fold higher sensitivity to the inhibitory activity of these CBAs was observed. Our data may provide an explanation why glycan deletions in the gp120 V1/V2 domain rarely occur under CBA pressure and confirm the important functional role of the glycans in the HIV-1 gp120 V1/V2 domain. The gp120 V1/V2 loop glycans of HIV-1 should therefore be considered as a hot spot and novel target for specific therapeutic drug intervention. | 18,930,512 |
Emergence and control of infectious diseases in China. | Infectious diseases remain the major causes of morbidity and mortality in China despite substantial progress in their control. China is a major contributor to the worldwide infectious disease burden because of its population size. The association of China with the rest of the world through travel and trade means that events in the country can affect distant populations. The ecological interaction of people with animals in China favours the emergence of new microbial threats. The public-health system has to be prepared to deal with the challenges of newly emerging infectious diseases and at the same time try to control existing diseases. To address the microbial threats, such as severe acute respiratory syndrome, the government has committed substantial resources to the implementation of new strategies, including the development of a real-time monitoring system as part of the infectious-disease surveillance. This strategy can serve as a model for worldwide surveillance and response to threats from infectious diseases. | 18,930,534 |
Analysis of B-cell epitopes from the allergen Hev b 6.02 revealed by using blocking antibodies. | Hev b 6.02 (hevein), identified as a major allergen from natural rubber latex (NRL), is involved in the latex-fruit syndrome and also acts as a pathogenesis defense-related protein. Its 3D structure has been solved at high resolution, and its linear epitopes have already been reported. However, information about conformational epitopes is still controversial, even though it is relevant for an accurate diagnosis and treatment, as well as for the study of allergen-antibody molecular interactions. We sought to analyze the B-cell epitopes of Hev b 6.02 at a molecular and structural level, using specific recombinant antibodies. We obtained a murine monoclonal antibody (mAb 6E7) and three human single chain fragments (scFvs A6, H8, and G7) anti-Hev b 6.02 that were able to compete for hevein binding with serum IgEs from latex allergic patients. In vitro assays showed that the mAb 6E7 and scFv H8 recognized the area of Hev b 6.02 where the aromatic residues are exposed; while the scFv G7 defined the amino and carboxy-terminal regions that lie close to each other, as a different epitope. The structural modeling of the Hev b 6.02-scFv H8 and Hev b 6.02-scFv G7 complexes revealed the putative regions of two conformational epitopes. In one of these, the aromatic residues, as well as polar side chains are important for the interaction, suggesting that they are part of a dominant conformational epitope also presented on the Hev b 6.02-IgE interactions. Antibodies recognizing this important allergen have potential to be used to diagnose and ultimately treat latex allergy. | 18,930,549 |
Treatment of ocular mucous membrane pemphigoid with immunosuppressive drug therapy. | To evaluate the effectiveness of immunosuppressive drug therapy in the treatment of ocular mucous membrane pemphigoid (MMP). Retrospective cohort study. Ninety-four patients with biopsy-proven ocular MMP seen at the Pemphigoid Clinic at Wilmer Eye Institute from July 1984 through November 2006. Data recorded included demographics, use and doses of immunosuppressive drugs, response to therapy, and side effects associated with drug use. Outcome measures included: (1) ocular control, defined as resolution of inflammation and cessation of cicatrization of the conjunctiva; (2) ocular remission, defined as ocular control for 3 months or more after the cessation of immunosuppressive drug therapy; and (3) ocular relapse, defined as the recurrence of ocular disease in either eye after a remission. By 1 year of treatment, 82.9% of patients had complete control of the inflammation, and of these, 86.3% achieved a remission at some point during follow-up. The incidences of ocular control, remission, and relapse were 1.03 (95% confidence interval [CI], 0.78-1.33), 0.50 (95% CI, 0.37-0.67), and 0.04 (95% CI, 0.02-0.09) events per person-years (PY), respectively. Among patients initially treated with prednisone and cyclophosphamide (n = 44), 91% of patients achieved a remission within 2 years after the initiation of immunosuppressive drug therapy. Characteristics at presentation associated with failing to achieve remission in the univariate analysis were trichiasis (relative risk [RR], 0.28; 95% CI, 0.08-097), prior eyelid surgery (RR, 0.11; 95% CI, 0.02-0.78), and esophageal involvement (RR, 0.29; 95% CI, 0.10-0.83). After adjusting for confounding, an initial treatment regimen containing cyclophosphamide and prednisone was associated with a greater likelihood of achieving ocular remission (RR, 8.53; 95% CI, 2.53-28.86; P = 0.001) when compared with other initial treatment regimens. Infections, hematuria, and anemia were the most common side effects observed in patients receiving cyclophosphamide therapy. The rate of discontinuing cyclophosphamide resulting from side effects was 0.20/PY; however, 74% of these patients still achieved remission despite early discontinuation of cyclophosphamide. In patients with ocular MMP, most achieved ocular disease control with immunosuppressive drug therapy. Treatment with cyclophosphamide and prednisone was strongly associated with the development of ocular remission. The author(s) have no proprietary or commercial interest in any materials discussed in this article. | 18,930,554 |
Computerized model of cost-utility analysis for treatment of age-related macular degeneration. | To present a computerized model assessing individualized cost utility for current treatments for neovascular age-related macular degeneration (AMD) to enhance discussion regarding treatment options. Case- and eye-specific cost-utility analysis using individual case scenarios. Visual acuity data from published randomized controlled trials are incorporated into this analysis. Computerized model (Microsoft Visual Basic 6.0 programming) to establish preference-based cost-utility analysis in association with individual cost of treatment and blindness for neovascular AMD for both the better and worst seeing eye, with extrapolation of results over a 5-year term. Cost per quality-adjusted life-year (QALY) and cost per QALY gained for comparison of treatments for specific visual acuities. All treatments show an increase in utility in comparison with best supportive care (BSC) if the better-seeing eye is treated. Ranibizumab, using the Phase IIIb, Multicenter, Randomized, Double-Masked, Sham Injection-Controlled Study of the Efficacy and Safety of Ranibizumab in Subjects with Subfoveal Choroidal Neovascularisation (CNV) with or without Classic CNV Secondary to AMD (PIER) regimen, is the most cost effective at $626 938 per QALY gained for treatment of the better seeing eye. To increase utility value when treating the worst seeing eye, the vision must improve to such a degree that it becomes the better seeing eye. This level of improvement is only possible if there is <9 letters difference between the 2 eyes and treated with ranibizumab. Over 5 years, increasing influence from the cost of blindness results in increasing costs for those treatments unable to stabilize vision. Within 5 years, the cost per QALY for the BSC is greater than all treatments except monthly ranibizumab injections. Assessment of cost of treatment incorporates both effectiveness of treatment, cost of treatment, and cost of blindness. Cost analysis enables incorporation of these aspects of treatment with the quality of life data to provide a better comparison of treatments over time. This analysis has provided a method for individual analysis and therefore can provide the structure for resource allocation. The authors have no proprietary or commercial interest in any materials discussed in this article. | 18,930,556 |
Combination therapy of prednisone and ACE inhibitor versus ACE-inhibitor therapy alone in patients with IgA nephropathy: a randomized controlled trial. | Recent studies have shown that both steroids and angiotensin-converting enzyme (ACE) inhibitors improve kidney survival and decrease proteinuria in patients with immunoglobulin A nephropathy. In this study, we aim to investigate whether the addition of steroids to ACE-inhibitor therapy produces a more potent antiproteinuric effect and better protection of kidney function than an ACE inhibitor alone. Randomized controlled trial. Patients with biopsy-proven immunoglobulin A nephropathy with proteinuria of 1 to 5 g/d of protein. 63 patients were randomly assigned to either cilazapril alone (ACE-inhibitor group; n = 30) or steroid plus cilazapril (combination group; n = 33). The primary end point was kidney survival, defined as a 50% increase in baseline serum creatinine level. After follow-up for up to 48 months, 7 patients in the ACE-inhibitor group (24.1%) reached the primary end point compared with 1 patient (3%) in the combination group. Kaplan-Meier kidney survival was significantly better in the combination group than the ACE-inhibitor group after 24 and 36 months (96.6% versus 75.7%, 96.6% versus 66.2%; P = 0.001). Urine protein excretion significantly decreased in patients in the combination group compared with the ACE-inhibitor group (time-average proteinuria, 1.04 +/- 0.54 versus 1.57 +/- 0.86 g/d of protein; P = 0.01). Multivariate analysis showed that combination treatment (hazard ratio, 0.1; 95% confidence interval, 0.014 to 0.946) and time-average proteinuria (hazard ratio, 14.3; 95% confidence interval, 2.86 to 71.92) were independent predictors of kidney survival. Small sample size, a single center, and slight imbalances at baseline. Our results suggest that the addition of steroid to ACE-inhibitor therapy provided additional benefit compared with an ACE inhibitor alone. However, this was a pilot study with a small number of participants achieving the end points, and thus further validation is necessary. | 18,930,568 |
Social assets and mental distress among the homeless: exploring the roles of social support and other forms of social capital on depression. | This paper explores the role of social capital in mediating the effects of stressors on depression among a disadvantaged population. Utilizing a survey of 155 homeless people in a mid-sized southern U.S. city, the authors address the relevance of social capital for quality of life. The paper provides a critical test of whether social support and other forms of social capital matter when monetary and human capital is extremely limited. Under these resource-restricted circumstances does social capital add to our understanding of the distress process or does it merely restate the well established relationship between social support and quality of life outcomes? Various forms of social capital are measured: religious social capital, group participation, social trust, and bridging social capital along with a commonly used measure of social support -- perceived strong tie support. Findings suggest that social capital matters for even the most resource poor populations. In addition, social capital variables add significantly to the variance explained in depressive symptomatology over and above that traditionally explained by perceived social support. | 18,930,571 |
Applications of Brazilian pine-fruit shell in natural and carbonized forms as adsorbents to removal of methylene blue from aqueous solutions--kinetic and equilibrium study. | The Brazilian pine-fruit shell (Araucaria angustifolia) is a food residue, which was used in natural and carbonized forms, as low-cost adsorbents for the removal of methylene blue (MB) from aqueous solutions. Chemical treatment of Brazilian pine-fruit shell (PW), with sulfuric acid produced a non-activated carbonaceous material (C-PW). Both PW and C-PW were tested as low-cost adsorbents for the removal of MB from aqueous effluents. It was observed that C-PW leaded to a remarkable increase in the specific surface area, average porous volume, and average porous diameter of the adsorbent when compared to PW. The effects of shaking time, adsorbent dosage and pH on adsorption capacity were studied. In basic pH region (pH 8.5) the adsorption of MB was favorable. The contact time required to obtain the equilibrium was 6 and 4h at 25 degrees C, using PW and C-PW as adsorbents, respectively. Based on error function values (F(error)) the kinetic data were better fitted to fractionary-order kinetic model when compared to pseudo-first order, pseudo-second order, and chemisorption kinetic models. The equilibrium data were fitted to Langmuir, Freundlich, Sips and Redlich-Peterson isotherm models. For MB dye the equilibrium data were better fitted to the Sips isotherm model using PW and C-PW as adsorbents. | 18,930,589 |
OmpA and antigenic diversity of bovine Chlamydophila pecorum strains. | Infections with the intracellular bacterium Chlamydophila (C.) pecorum are highly prevalent worldwide in cattle. These infections cause significant diseases such as polyarthritis, pneumonia, enteritis, genital infections and fertility disorders, and occasionally sporadic bovine encephalomyelitis. Subclinical respiratory infections of calves with C. pecorum have been associated with airway obstruction, pulmonary inflammation, and reduced weight gains. This investigation examined four chlamydial strains with biological properties of C. pecorum isolated from feces of clinically normal cattle, from calves with pneumonia, and from bulls with posthitis. The objective was to characterize the evolutionary relationships of these bovine chlamydial isolates to other chlamydiae by genetic analysis of the ompA gene, and by the immunological cross-reactivities in Western immunoblot analysis. PCR typing of the ompA gene identified these isolates as C. pecorum. The OmpA-deduced amino acid dissimilarities between these four strains spanned 10-20%. In phylogenetic analysis, the four isolates clustered with C. pecorum ruminant, porcine, and koala strains of different geographic origins rather than with each other. All four isolates showed different patterns of Western immunoblot reactivity with antiserum against bovine C. pecorum strain LW63, and, interestingly, no cross-reactivity of the OmpA proteins with the anti-LW613 OmpA antibodies. These data underscore the polyphyletic population structure of C. pecorum and suggest that the spectrum of C. pecorum OmpA proteins in a host species can occupy the entire evolutionary bandwidth within C. pecorum. The variant immunoblot reactivities support the notion of considerable genomic plasticity of C. pecorum. | 18,930,605 |
Molecular detection of Chlamydophila abortus in post-abortion sheep at oestrus and subsequent lambing. | Enzootic abortion of ewes (EAE), resulting from infection with the bacterium Chlamydophila abortus (C. abortus), is a major cause of lamb loss in Europe. The purpose of this study was to assess the potential impact of the shedding of organisms in post-abortion ewes at oestrus and subsequent lambing on the epidemiology of EAE. Using a newly developed C. abortus specific real-time PCR assay, few chlamydial genomes could be detected in vaginal swabs taken from post-abortion ewes at oestrus. At subsequent parturition, all ewes lambed normally with no macroscopic or microbiological evidence of infection. Real-time PCR analysis of placental samples identified very few or no chlamydial genomes, which contrasted significantly with samples taken at the time of abortion, where an average of 2.7x10(7) chlamydial genomes per microgram of total tissue DNA was detected. Few genomes could also be detected from vaginal and cervical tissue samples and lymph nodes taken post-mortem. The results, although not discounting the possibility of a chronic low level persistent infection in post-abortion ewes, suggest that the low levels of chlamydial DNA detected during the periovulation period and at lambing do not significantly impact on the epidemiology of EAE. In terms of flock management, the products of abortion should be considered the major and principal source of infection for transmission to naïve ewes. | 18,930,611 |
Clinical utility of contrast-enhanced three-dimensional ultrasound imaging with Sonazoid: findings on hepatocellular carcinoma lesions. | We investigated clinical utility of contrast-enhanced three-dimensional ultrasound (CE 3D US) imaging with contrast medium Sonazoid for demonstrating characteristic enhancement of hepatocellular carcinoma (HCC). Among 115 focal liver tumors undergoing CE 3D US imaging, 70 HCCs confirmed with contrast-enhanced multi-detector computed tomography, contrast-enhanced magnetic resonance imaging or histopathologic examination were retrospectively analyzed. CE 3D US imaging was performed using Autosweep 3D scan functionality in the early, middle and late phase, after bolus injection of 0.2 ml Sonazoid. The CE 3D tomographic images reconstructed in parallel slices and sonographic angiogram images were independently reviewed by two reviewers. Kappa values were used to assess inter-reviewers' agreement. TUI images showed most of HCCs were detected with intratumoral vessels and early tumor enhancement in the early phase, expressed homogenous or heterogeneous tumor enhancement in the middle phase, and became hypoechoic or isoechoic in the late phase. The kappa values in the early, middle and late phase for inter-reviewer agreements regarding the characteristic enhancement of tumors were 0.817, 0.774, and 0.785. In addition, TUI images demonstrated satellite foci and tortuous tumor vessels in three orthogonal planes. Sonographic angiogram reconstructed by different rendering modes showed the vessels and tumor stain in spatial view. The spatial configuration of anatomic structures was revealed on basis of both TUI and sonographic angiogram images. CE 3D US imaging, with spatial visualization, is clinically useful to exhibit the characteristic enhancement of HCC tumors objectively. | 18,930,616 |
Choice of reference area in studies of Alzheimer's disease using positron emission tomography with fluorodeoxyglucose-F18. | At present, there is still no consensus on the choice of the reference area in positron emission tomography (PET) studies of Alzheimer's disease (AD). In this study, PET scans with fluorodeoxyglucose-F18 were carried out in the following groups of subjects: 47 patients with probable AD, 8 patients with mild cognitive impairment, and 15 age-similar healthy subjects. Scans normalized to the cerebral global mean (CGM), cerebellum (CBL), and the primary sensorimotor cortex (SMC). We evaluated the effect of the different count normalization procedures on the accuracy of (18)F-FDG PET to detect AD-specific metabolic abnormalities (voxel-based group comparison) and to differentiate between patients and healthy subjects (ROI-based discriminant analysis) with regard to the degree of clinical deterioration. Metabolic reductions in groups of very mildly, mildly and moderate-to-severely affected patients appeared, respectively, 2.2, 2.6, and 2.7 times greater in spatial extent when tracer uptake was normalized to SMC rather than to CGM. The overall accuracy of discrimination was 94%, 91%, and 80% after normalization to SMC, CBL, and CGM, respectively. In general, normalization to SMC was somewhat superior to cerebellar normalization, allowing the detection of more pronounced metabolic deficits and the more accurate discrimination of patients from non-patients. Normalization to CGM should be used with great caution not only in advanced stages of dementia, but also in very mild AD cases. | 18,930,634 |
Positron emission tomography with fluorodeoxyglucose-F18 in an animal model of mania. | Intracerebroventricular (ICV) administration of ouabain to young adult rats has been suggested to model human bipolar mania. In the human condition, mania and bipolar depression are both associated with reductions in frontal cerebral metabolism. We utilized [(18)F]-fluorodeoxyglucose [(18)FDG] positron emission tomography (PET) to visualize glucose uptake in animals receiving ICV ouabain. Animals received 5 microl of 10(-)(3) M ouabain ICV, were anesthetized with isoflurane inhalation, and administered intraperitoneally with 0.5 mCi of (18)FDG. PET data were collected over 20 min 1 hour later. Additionally, the effect of lithium was examined in animals receiving lithium in their diet for 1 week before the ICV ouabain injection. Data were analyzed with IDL Virtual Machine software. Brain glucose utilization as measured by (18)FDG uptake was significantly reduced in animals receiving ICV ouabain compared with those receiving equal volumes of artificial cerebrospinal fluid. Pretreatment with lithium normalized (18)FDG uptake. These results mirror human studies. | 18,930,636 |
The lipidome as a composite biomarker of the modifiable part of the risk of breast cancer. | The potential for dietary fat to prevent breast cancer makes identification of defined molecules a mandatory step. In order to circumvent the limitations and/or bias of dietary exposure assessment tools, we have used the fatty acid composition of white adipose tissue as biomarker of past lipid intake. When considered separately, candidate fatty acids identified as favourable on the basis of their association with breast cancer risk have usually led to inconsistent results in dietary intervention studies carried out in rats. This inconsistency indicates that any approach based on a single fatty acid should be abandoned for an integrated view over the complex lipid interactions, which finally determines the lipidome, the lipid profile that is found in individuals. We reappraised the role of the complete lipid profile through a comprehensive study of adipose tissue fatty acids obtained in patients with benign or malignant breast tumors. Rather than a single fatty acid, a composite indicator combining elevated monounsaturates and low n-6/n-3 fatty acid ratio was associated with decreased breast cancer risk. The lipidome may provide the opportunity to quantify the modifiable part of the risk of breast cancer. The lipidome may be used as a template for designing proper dietary modifications in order to delay the occurrence of breast cancer. Which dietary modifications should be undertaken in order to bring a pertinent change to the lipidome with respect to the risk of breast cancer is currently unknown. The lipidome may allow the individualization of a high risk population of women, who may be targeted for a dietary prevention of breast cancer. The setting and validation of a high-throughput lipidomic station with analytical capabilities fitted to the need of mass screening is required. These two locks must be resolved before a primary prevention of breast cancer by diet could be contemplated. | 18,930,643 |
A technological advance comparing epithelial lining fluid from different regions of the lung in smokers. | Cigarette smoking causes inflammatory responses in the airways. However, not all smokers exhibit the development of airflow limitation. This study was designed to determine the implications of small airways inflammation in the development of airflow limitation in smokers by our newly explored method. Twenty-eight smokers (15 smokers without airflow limitation and 13 with airflow limitation) were included in this study. Levels of interleukin-8 (IL-8) and 8-isoprostane were measured in epithelial lining fluid (ELF) from central and peripheral airways separately collected using a bronchoscopic microsampling technique. 8-isoprostane levels in ELF from central or peripheral airways did not significantly differ between the two groups. However, these levels were markedly higher in peripheral than in central airways. Similarly, IL-8 levels in ELF from central airways did not significantly differ between the two groups. In smokers without airflow limitation, IL-8 levels were not higher in peripheral than in central airways. In contrast, in smokers with airflow limitation, IL-8 levels were significantly higher in peripheral airways. Moreover, in smokers with airflow limitation, 8-isoprostane levels in central or peripheral airways were not significantly correlated with FEV(1). However, IL-8 levels in peripheral airways were inversely correlated with FEV(1), though those levels in central airways were not. Thus our technique provides a novel method for ELF sampling from central or peripheral airways separately, and the preliminary evidence that support differences in oxidative stress and neutrophil chemotactic stimulus in these two locations. | 18,930,648 |
Social epidemiology and complex system dynamic modelling as applied to health behaviour and drug use research. | A social epidemiologic perspective considers factors at multiple levels of influence (e.g., social networks, neighbourhoods, states) that may individually or jointly affect health and health behaviour. This provides a useful lens through which to understand the production of health behaviours in general, and drug use in particular. However, the analytic models that are commonly applied in population health sciences limit the inference we are able to draw about the determination of health behaviour by factors, likely interrelated, across levels of influence. Complex system dynamic modelling techniques may be useful in enabling the adoption of a social epidemiologic approach in health behaviour and drug use research. We provide an example of a model that aims to incorporate factors at multiple levels of influence in understanding drug dependence. We conclude with suggestions about future directions in the field and how such models may serve as virtual laboratories for policy experiments aimed at improving health behaviour. | 18,930,649 |
Evaluation of flow scintillation analysis for the determination of Sr-90 in bioassay samples. | An automated procedure for the determination of (90)Sr was adapted from existing methods of flow scintillation analysis (FSA) for use on aqueous samples with low levels of activity (<1000 dpm per sample). This technique employs high-performance extraction chromatography (HPEC) and an on-line liquid scintillation counter to provide automated separation and subsequent detection of (90)Sr. The total analysis time is 30 min per sample. Dilute urine samples, spiked with (90)Sr, were also processed by this method to test the application of this technique for bioassay monitoring. | 18,930,656 |
Intralesional laser treatment of voluminous vascular lesions in the oral cavity. | In laser treatment of voluminous vascular lesions, there are many cases in which submucosally located angioma remnants cannot be reached by noncontact superficial laser application. To diminish these remnants we used intralesional photocoagulation (ILP) in treatment of oral vascular lesions, because this approach is effective in treatment of voluminous vascular lesions of the skin. Four cases of voluminous vascular malformation in the oral cavity were treated by ILP using a potassium-titanyl-phosphate (KTP) laser. In 1 case, treatment was carried out under ultrasound and manual control. All lesions showed more than 70% regression after the first ILP session, and the treatment outcome was satisfactory. There were no serious complications, such as bleeding or invasive infection. Ultrasonography was useful for guiding laser treatment in the oral cavity. Intralesional photocoagulation treatment with a KTP laser is effective and safe for treatment of a vascular lesion in the oral cavity. | 18,930,663 |
Endotoxin concentrations within the breathing zone of horses are higher in stables than on pasture. | Inflammatory airway disease is common in stabled horses, with a prevalence of 17.3% in Michigan pleasure horses. Stable dust is rich in endotoxin, which may induce neutrophilic airway inflammation. Climatological conditions (ambient temperature and relative humidity) may influence endotoxin concentrations in pastures. The aim of this project was to determine if endotoxin levels in the breathing zone of horses in stables were higher than of horses on pasture, and if the endotoxin on pasture was associated with climatological conditions. Endotoxin exposure of six horses that were stabled or on pasture was determined by a Limulus amebocyte lysate assay. Climatological data were obtained from the US National Climatic Data Center. Endotoxin exposure was significantly higher (about 8-fold) in stables than on pasture. On pasture, endotoxin varied widely, despite constant climatological conditions. It was concluded that stabled horses are exposed to higher endotoxin concentrations than horses on pastures. Local endotoxin concentrations may be more important than ambient climatological conditions in determining endotoxin exposure of individual horses. | 18,930,664 |
A novel form of aberrant innervation in congenital cranial dysinnervation disorder. | Certain forms of congenital incomitant strabismus are associated with abnormal cranial nerve development and resultant abnormal orbital innervation (eg, Duane retraction syndrome, congenital fibrosis of the extraocular muscles [CFEOM]); such conditions can be considered congenital cranial dysinnervation disorders (CCDDs). In addition to duction limitation and/or ptosis, orbital CCDD phenotypes include inappropriate extraocular muscle and/or levator innervation by nerves intended for innervation of other structures (eg, some of the innervation intended for the medial rectus muscle inappropriately innervating the ipsilateral lateral rectus muscle in Duane retraction syndrome). This report documents a unique orbital dysinnervational pattern-supraduction during attempted adduction and infraduction during attempted abduction in the left affected eye of a girl with exotropia and enophthalmos. | 18,930,669 |
Improvement with ongoing Enzyme Replacement Therapy in advanced late-onset Pompe disease: a case study. | Benefits of enzyme replacement therapy with Myozyme (alglucosidase alfa), anecdotally reported in late-onset Pompe disease, range from motor and pulmonary improvement in less severely affected patients, to stabilization with minimal improvement in those with advanced disease. We report a case of a 63-year-old patient with significant morbidity who made notable motor and pulmonary function gains after two years on therapy. Thus, improvements in those with advanced disease may be possible after long-term treatment. | 18,930,676 |
Mediterranean spotted fever in Algeria--new trends. | Mediterranean spotted fever (MSF) due to Rickettsia conorii is the most important tick-borne disease occurring in North Africa. However, there are only a few fragmentary reports on the epidemiology and clinical aspects of rickettsioses in North Africa, and cases are still rarely documented. We report herein a prospective study conducted in Oran, the second largest city in Algeria. This disease has not been properly described in Oran or in other Algerian cities. A total of 167 cases of Mediterranean spotted fever were documented for the first time by the use of reference methods including immunofluorescence serology and Western blot and absorption studies, including isolation in culture by the shell-vial techniques, and molecular tools. Although some aspects of MSF were found to be in accordance with the general epidemiology of the disease, uncommon aspects were found, including increased incidence and the presence of multiple inoculation eschars in 12% of patients. The role of climatic changes in alterations of host-seeking and feeding behaviors of the vectors, including the brown dog tick Rhipicephalus sanguineus, is discussed. Also, 49% of patients were hospitalized with a severe form. The global death rate was 3.6%, but it was 54.5% in patients hospitalized with major neurological manifestations and multiorgan involvement. The present report gives a unique panel of clinical aspects of MSF as well as new trends in this disease. Entomological, climatic, and molecular studies are needed to better understand both epidemiological and clinical aspects of MSF. | 18,930,677 |
The cytochrome ba complex from the thermoacidophilic crenarchaeote Acidianus ambivalens is an analog of bc(1) complexes. | A novel cytochrome ba complex was isolated from aerobically grown cells of the thermoacidophilic archaeon Acidianus ambivalens. The complex was purified with two subunits, which are encoded by the cbsA and soxN genes. These genes are part of the pentacistronic cbsAB-soxLN-odsN locus. The spectroscopic characterization revealed the presence of three low-spin hemes, two of the b and one of the a(s)-type with reduction potentials of +200, +400 and +160 mV, respectively. The SoxN protein is proposed to harbor the heme b of lower reduction potential and the heme a(s), and CbsA the other heme b. The soxL gene encodes a Rieske protein, which was expressed in E. coli; its reduction potential was determined to be +320 mV. Topology predictions showed that SoxN, CbsB and CbsA should contain 12, 9 and one transmembrane alpha-helices, respectively, with SoxN having a predicted fold very similar to those of the cytochromes b in bc(1) complexes. The presence of two quinol binding motifs was also predicted in SoxN. Based on these findings, we propose that the A. ambivalens cytochrome ba complex is analogous to the bc(1) complexes of bacteria and mitochondria, however with distinct subunits and heme types. | 18,930,705 |
A pharmacological comparison of the cloned frog and human mu opioid receptors reveals differences in opioid affinity and function. | This study presents a direct comparison of the ligand binding and signaling profiles of a mammalian and non-mammalian mu opioid receptor. Opioid ligand binding and agonist potencies were determined for an amphibian (Rana pipiens) mu opioid receptor (rpMOR) and the human mu opioid receptor (hMOR) in transfected, intact Chinese hamster ovary (CHO) cells. Identical conditions were employed such that statistically meaningful differences between the two receptors could be determined. Identifying these differences is an important first step in understanding how evolutionary changes affect ligand binding and signaling in vertebrate opioid receptors. As expected, the rank of opioid ligand affinity for rpMOR and hMOR was consistent with the ligands' previously characterized type-selectivity. However, most of the opioid ligands tested had significant differences in affinity for rpMOR and hMOR. For example, the mu-selective agonist, DAMGO ([d-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin), had a 10.9-fold greater affinity (K(i)) for hMOR (K(i)=268 nM) than rpMOR (K(i)=2914 nM). In addition, differences in signaling between these receptors were found by measuring inhibition of cAMP accumulation by morphine or DAMGO. DAMGO was significantly more potent (13.6-fold) in CHO cells expressing hMOR versus those expressing rpMOR. In addition, a significantly greater maximal inhibition was elicited by both opioid agonists in cells expressing hMOR. In summary, this study supports an ongoing effort to better understand how vertebrate evolution has shaped opioid receptor properties and function. | 18,930,720 |
The tachykinin NK3 receptor agonist senktide induces locomotor activity in male Mongolian gerbils. | The tachykinin family of receptors has been of strong interest recently due to the potential of the tachykinin NK(3) receptor antagonism in treatment of schizophrenia. However, critical differences in the tachykinin NK(3) receptor between rats, mice and humans make rats and mice less acceptable species for testing tachykinin NK(3) receptor antagonism. This has led to testing of tachykinin NK(3) receptor activity in gerbils and guinea pigs. As these species are much less common laboratory animals than rats and mice, there is a relative paucity of in vivo testing models for tachykinin NK(3) receptor activation. In the present study, locomotor activity induced by the tachykinin NK(3) receptor agonist senktide was characterized. Injection of senktide i.c.v. was found to dose-dependently induce hyperlocomotion from a dose of 0.06 nmol to the maximal dose tested, 0.6 nmol. Locomotion induced by 0.1 nmol of senktide could be blocked by injection of the tachykinin NK(3) receptor antagonists SB222200 (10 and 30 mg/kg i.p.) and talnetant (SB223412; 10 and 30 mg/kg i.p.), as well as by osanetant (SR142801; 10 and 30 mg/kg i.p.) when administered in a vehicle containing vitamin E and glycofurol. Senktide-induced activity was also reversed by the antipsychotics haloperidol (0.3 and 1 mg/kg p.o.) and risperidone (1 mg/kg p.o.), but not by the serotonin 5HT(2a/c) receptor antagonist MDL100907 (tested at 0.1, 0.3 and 1 mg/kg p.o.). Hyperlocomotion induced by 0.03 nmol of senktide was potentiated by antagonism of the tachykinin NK(1) receptor with aprepitant (1, 3 and 10 mg/kg, p.o.). Thus, hyperlocomotion induced by senktide in gerbils is a tachykinin NK(3) receptor-mediated behavior that is appropriate for use in testing tachykinin NK(3) receptor activity of novel compounds. | 18,930,726 |
Effect of xenon binding to a hydrophobic cavity on the proton pumping cycle in bacteriorhodopsin. | To understand the functional role of apolar cavities in bacteriorhodopsin, a light-driven proton pump found in Halobacterium salinarum, we investigated the crystal structure in pressurized xenon or krypton. Diffraction data from the P622 crystal showed that one Xe or Kr atom binds to a preexisting hydrophobic cavity buried between helices C and D, located at the same depth from the membrane surface as Asp96, a key residue in the proton uptake pathway. The occupation fraction of Xe or Kr was calculated as approximately 0.32 at a pressure of 1 MPa. In the unphotolyzed state, the binding of Xe or Kr caused no large deformation of the cavity. However, the proton pumping cycle was greatly perturbed when an aqueous suspension of purple membrane was pressurized with xenon gas; that is, the decay of the M state was accelerated significantly (~5 times at full occupancy), while the decay of an equilibrium state of N and O was slightly decelerated. A similar but much smaller perturbation in the reaction kinetics was observed upon pressurization with krypton gas. In a glycerol/water mixture, xenon-induced acceleration of M decay became less significant in proportion to the water activity. Together with the structure of the xenon-bound protein, these observations suggest that xenon binding helps water molecules permeate into apolar cavities in the proton uptake pathway, thereby accelerating the water-mediated proton transfer from Asp96 to the Schiff base. | 18,930,734 |
Excessive S-adenosyl-L-methionine-dependent methylation increases levels of methanol, formaldehyde and formic acid in rat brain striatal homogenates: possible role in S-adenosyl-L-methionine-induced Parkinson's disease-like disorders. | Excessive methylation may be a precipitating factor for Parkinson's disease (PD) since S-adenosylmethionine (SAM), the endogenous methyl donor, induces PD-like changes when injected into the rat brain. The hydrolysis of the methyl ester bond of the methylated proteins produces methanol. Since methanol is oxidized into formaldehyde, and formaldehyde into formic acid in the body, we investigated the effects of SAM on the production of methanol, formaldehyde and formic acid in rat brain striatal homogenates and the toxicity of these products in PC12 cells. Radio-enzymatic and colorimetric assays, cell viability, Western blot. SAM increased the formation of methanol, formaldehyde and formic acid in a concentration and time-dependent manner. Concentrations of [3H-methyl]-SAM at 0.17, 0.33, 0.67 and 1.34 nM produced 3.8, 8.0, 18.3 and 34.4 fmol/mg protein/h of [3H] methanol in rat striatal homogenates, respectively. SAM also significantly generated formaldehyde and formic acid in striatal homogenates. Formaldehyde was the most toxic metabolite to differentiated PC12 pheochromocytoma cells in cell culture studies, indicating that formaldehyde formed endogenously may contribute to neuronal damage in excessive methylation conditions. Subtoxic concentration of formaldehyde decreased the expression of tyrosine hydroxylase, the limiting factor in dopamine synthesis. Formaldehyde was more toxic to catecholaminergic PC12 cells than C6 glioma cells, indicating that neurons are more vulnerable to formaldehyde than glia cells. We suggest that excessive carboxylmethylation of proteins might be involved in the SAM-induced PD-like changes and in the aging process via the toxic effects of formaldehyde. | 18,930,743 |
Dynamic models for the study of frailty. | Frailty can be viewed as resulting from the degradation of multiple interacting physiologic systems that are normally responsible for healthy adaptation to the daily demands of life. Mathematical models that can quantify alterations in the dynamics of physiologic systems and their interactions may help characterize the syndrome of frailty and enable investigators to test interventions to prevent its onset. One theoretical mathematical model reported by Varadhan et al. in this issue of the Journal represents one type of regulatory process that may become altered in frail individuals-the stimulus-response mechanism [Varadhan, R., Seplaki, C.S., Xue, Q.L., Bandeen-Roche, K., Fried, L.P. Stimulus-response paradigm for characterizing the loss of resilience in homeostatic regulation associated with frailty. Mech. Ageing Dev., this issue]. This model focuses on the timing of recovery from a single stimulus, rather than the full array of responses that might be altered in a complex dynamical system. Therefore, alternative models are needed to describe the wide variety of behaviors of physiologic systems over time and how they change with the onset of frailty. One such model, based on a simple signaling network composed of a lattice of nodes and the bi-directional connections between them, can reproduce the complex, fractal-like nature of healthy physiological processes. This model can be used to demonstrate how the degradation of signaling pathways within a physiologic system can result in the loss of complex dynamics that characterizes frailty. | 18,930,754 |
Construction of a new polycistronic vector for over-expression and rapid purification of human hemoglobin. | To facilitate the study of the structure-function relationship of human hemoglobin (Hb A), we have developed a new hemoglobin expression vector, pGEX6P-alpha-[SD]-beta. This vector allows the co-expression of alpha-Hb as a fusion protein with Glutathione S-Transferase (GST-alpha-Hb) and beta-Hb with an additional methionine at the N-terminal extremity (rbeta-Hb). These proteins were solubilized as GST-alpha-Hb/rbeta-Hb complex form and purified in one step by affinity chromatography on immobilized glutathione. The CO binding kinetic studies show that the GST-alpha-Hb/rbeta-Hb complex and recombinant Hb A exhibit the same allosteric behavior as for native Hb A. The GST moiety, which does not modify the function of the complex, can be easily eliminated by cleavage by the PreScission Protease. After cleavage during the rapid purification procedure, over 20mg of recombinant Hb per liter of culture were obtained, more than double the yield of previous co-expression systems. This polycistronic vector system, which offers the additional advantage of a very rapid purification, is especially well suited for the study of abnormal, unstable globins in order to better understand the associated pathology. | 18,930,760 |
Prophylactic neuroprotective property of Centella asiatica against 3-nitropropionic acid induced oxidative stress and mitochondrial dysfunctions in brain regions of prepubertal mice. | Despite the increasing popularity of Centella asiatica (a well known plant in ayurvedic medicine) globally, evidence demonstrating its protective efficacy against neurotoxicants in animal models is limited. 3-Nitropropionic acid (3-NPA), a fungal toxin is a well known neurotoxicant which induces selective striatal pathology similar to that seen in Huntington's disease. The present study aimed to understand the neuroprotective efficacy of a standardized aqueous extract of C. asiatica (CA) against 3-NPA-induced early oxidative stress and mitochondrial dysfunctions in striatum and other brain regions. We determined the extent of oxidative stress in cytosol and mitochondria of brain regions of male mice (4wk old) given CA prophylaxis (5mg/kgbw) for 10 days followed by 3-NPA administration (i.p., 75mg/kgbw/d) on the last 2 days. The neurotoxicant elicited marked oxidative stress in the untreated mice as evidenced by elevated levels of malondialdehyde, ROS levels and hydroperoxides in the striatum (cytosol and mitochondria), while CA prophylaxis completely attenuated the 3-NPA-induced oxidative stress. 3-NPA also caused significant oxidative stress and protein oxidation in cytosol/mitochondria of other brain regions as well which were predominantly abolished by CA prophylaxis. Significant depletion of GSH levels, total thiols and perturbations in antioxidant enzymic defences in striatum and other brain regions discernible among 3-NPA administered mice were also protected with CA prophylaxis. Interestingly, CA prophylaxis offered varying degree of protection against 3-NPA-induced mitochondrial dysfunctions viz., reduction in the activity of succinic dehydrogenase, ETC enzymes and decreased mitochondrial viability. Collectively these findings clearly suggest that short-term oral intake of a standardized aqueous extract of CA confers marked resistance against the 3-NPA-induced oxidative stress and mitochondrial dysfunctions in brain. Although the precise mechanism/s underlying the prophylactic efficacy of CA merit further investigation, based on these findings, it is hypothesized that it may be wholly or in part related to the enhancement of GSH, thiols and antioxidant machinery in the brain regions of prepubertal mice. | 18,930,762 |
A novel concept in enteric coating: a double-coating system providing rapid drug release in the proximal small intestine. | A novel double-coating enteric system was developed to accelerate drug release in conditions resembling the upper small intestine. The system comprises an inner coat (partially neutralised EUDRAGIT L 30 D-55 and organic acid) and an outer coat (standard EUDRAGIT L 30 D-55). Prednisolone tablets were coated with double layer formulations with inner coats neutralised to pH 5.6 in the presence of 10% citric acid or adipic acid. A conventional single coating was also applied for comparison purposes. There was no drug release from the single coated or double-coated tablets in 0.1M HCl for 2 h using USP II apparatus. The lag times of drug release in subsequent pH 5.6 phosphate buffer (to resemble the pH condition of the proximal small intestine) were 102, 42 and 28 min for the single coated, adipic acid and citric acid double-coated tablets respectively. The lag time for release from the double-coated tablets was further reduced to 5 min when the inner coat was neutralised to pH 6.0 in the presence of 10% citric acid. The rapid drug release from the double-coating system was associated with faster polymer dissolution rates compared to the single coating. The novel double-coated system has the potential to provide rapid drug release in the proximal small intestine, overcoming the limitations of conventional enteric coatings. | 18,930,772 |
The protective effect of erdosteine on short-term global brain ischemia/reperfusion injury in rats. | Experimental studies have demonstrated that free radicals play a major role on neuronal injury during ischemia/reperfusion (I/R) in rats. Erdosteine is a thioderivative endowed with mucokinetic, mucolytic and free-radical-scavenging properties. The aim of the present study was to investigate the effect of erdosteine treatment against short-term global brain ischemia/reperfusion injury in rats. The study was carried out on Wistar rats divided into four groups. (i) Control group, (ii) ischemia/reperfusion group, (iii) ischemia/reperfusion+erdosteine group, and (iv) erdosteine group. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities as well as thiobarbituric acid reactive substances (TBARSs) and nitric oxide (NO) levels were analysed in erythrocyte and plasma of rats. Plasma NO levels were significantly higher in the ischemia/reperfusion group than the other groups. The activities of SOD and GSH-Px were decreased, while TBARS levels increased in the ischemia/reperfusion group compared to other groups in both plasma and erythrocyte. The erythrocyte CAT activity was higher in erdosteine group and there was a statistically significant increase, when compared with the erdosteine plus ischemia/reperfusion group. By treating the rats with erdosteine, the depletion of endogenous antioxidant enzymes (SOD, CAT, GSH-Px) and increase of TBARS and NO levels were prevented. This study, therefore, suggests that erdosteine reduces parameters of oxidative stress is well supported by the data. | 18,930,779 |
D-Dopachrome tautomerase is a candidate for key proteins to protect the rat liver damaged by carbon tetrachloride. | Carbon tetrachloride (CCl4) is known to induce liver damage. Animal experiments with CCl4 injections have revealed many findings, especially mechanisms of liver damage and liver regeneration. Recently, proteomic approaches have been introduced in various studies to evaluate the quantitative and qualitative changes in the comprehensive proteome level. The aim of this research is to elucidate the key protein for liver damage, liver protection and liver regeneration by using proteomic techniques. 50 % (v/v) CCl4 in corn oil was administered intraperitoneally to adult male rats at a dose of 4ml/kg body weight. Approximately 24h after the injection, the liver was removed and extracted proteins were analyzed with cleavable isotope coded affinity tag (cICAT) reagents, two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). A twelvefold increase in D-dopachrome tautomerase (DDT) was indicated. This enzyme has been reported to be involved in the biosynthesis of melanin, an antioxidant. According to the histological analysis, melanin levels were increased in un-damaged hepatocytes of CCl4-treated rats. These results suggest that the increase in DDT is a response to liver damage, accelerates melanin biosynthesis and protects the liver from oxidative stress induced by CCl4. | 18,930,782 |
The inhibitory effect of diphenyltin on gap junctional intercellular communication in HEK-293 cells is reduced by thioredoxin reductase 1. | Organotins display high biological activity and are toxic to animals and humans. Besides carcinogenic effects, they have been shown to have highly immunotoxic and/or neurotoxic activity; however, the molecular mechanism of their toxicity is not fully understood. The ability of chemicals to inhibit communication via gap junctions has been associated with their toxicological properties. The aim of this study was to determine whether diphenyltin (DPhT) affects the gap junctional intercellular communication (GJIC) and whether thioredoxin reductase (TrxR1) is involved in the regulation of this process. We found that DPhT inhibits GJIC in HEK-293 cells. The inhibition of GJIC depends on the activation of PKC delta and is associated with the induction of Cx43 phosphorylation at Ser262. Moreover, we found that GJIC inhibited by DPhT in HEK-293 cells is fully re-established as a result of TrxR1 overexpression. | 18,930,794 |
Association between haplotypes of manganese superoxide dismutase (SOD2), smoking, and lung cancer risk. | Tobacco smoke contains high concentrations of reactive oxygen species (ROS) that can damage DNA, proteins, and lipids. Manganese superoxide dismutase (SOD2) catalyzes the dismutation of superoxide radicals into hydrogen peroxide and protects against oxidative stress in lung tissues. Three tagSNPs were identified in one block of high linkage disequilibrium that spans the entire SOD2 gene and 5-kb promoter region. These tagSNPs, representing four haplotypes (TAA, TCA, TCG, CCG), were genotyped in 372 lung cancer cases and 605 controls. There was no association between the haplotype frequencies and the overall lung cancer risk. The TCG haplotype (6% in controls) was significantly associated with a lower risk of lung cancer in light smokers (<median pack-years; P value=0.02) but not in heavy smokers. In histologic-specific analysis, the TCG haplotype was significantly associated with a reduced risk of lung adenocarcinoma (odds ratio=0.39, 95% CI 0.17-0.88), but an inverse association with squamous cell carcinoma was not significant. The association with adenocarcinoma was most apparent in light smokers (haplotype-specific P value=0.005); none of the 61 case subjects with adenocarcinoma had the TCG allele. This study suggests that subjects with the SOD2 TCG haplotype may be at decreased risk for lung adenocarcinoma and that this association may depend on smoking amount. | 18,930,810 |
The neuroprotective impact of the leak potassium channel TASK1 on stroke development in mice. | Oxygen depletion (O(2)) and a decrease in pH are initial pathophysiological events in stroke development, but secondary mechanisms of ischemic cell death are incompletely understood. By patch-clamp recordings of brain slice preparations we show that TASK1 and TASK3 channels are inhibited by pH-reduction (42+/-2%) and O(2) deprivation (36+/-5%) leading to membrane depolarization, increased input resistance and a switch in action potential generation under ischemic conditions. In vivo TASK blockade by anandamide significantly increased infarct volumes at 24 h in mice undergoing 30 min of transient middle cerebral artery occlusion (tMCAO). Moreover, blockade of TASK channels accelerated stroke development. Supporting these findings TASK1(-/-) mice developed significantly larger infarct volumes after tMCAO accompanied by worse outcome in functional neurological tests compared to wild type mice. In conclusion, our data provide evidence for an important role of functional TASK channels in limiting tissue damage during cerebral ischemia. | 18,930,826 |
Epidermal growth factor-induced ovarian carcinoma cell migration is associated with JAK2/STAT3 signals and changes in the abundance and localization of alpha6beta1 integrin. | Peritoneal dissemination of ovarian carcinoma is mediated by epithelial-mesenchymal interconversions leading to the disruption of cell-cell contact and modulation of cell-extracellular matrix (ECM) interactions. The present study was designed to evaluate the effects of epidermal growth factor (EGF) as a modulator of Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) signalling and changes in integrin expression during the process similar to EMT. A fibroblastic morphology with reduced intercellular cell contacts and increased cell motility was observed in ovarian cancer cell lines in response to EGF and was concomitant with the up regulation of EMT-associated N-cadherin and vimentin expression. These changes were accompanied by an increase in alpha2, alpha6 and beta1 integrin subunits and activation of JAK2 and STAT3 signalling which was suppressed by a specific JAK2 inhibitor. Consistent with the suppression of STAT3 activity, N-cadherin and vimentin expression were abrogated and was coherent with the loss of cell motility and the expression of alpha6 and beta1 integrin subunits. Neutralizing antibodies against alpha6 and beta1 subunits inhibited cancer cell migration. A strong correlation between the expression of N-cadherin, vimentin and JAK2/STAT3 levels were detected in high-grade ovarian tumors and was consistent with the previously reported enhanced expression of alpha6 integrin subunit in advanced tumors [Ahmed N, Riley C, Oliva K, Rice G, Quinn M. Ascites induces modulation of alpha6beta1 integrin and urokinase plasminogen activator receptor expression and associated functions in ovarian carcinoma. British Journal of Cancer 2005;92:1475-85]. Our data incorporating the clinical samples and the cancer cell lines is the first to demonstrate that JAK2/STAT3 pathway may be one of the downstream events in EMT-like process and alpha6beta1 integrin-mediated signalling in ovarian carcinomas. | 18,930,836 |
Chromosome aberration frequencies and chromosome instability in mice after long-term exposure to low-dose-rate gamma-irradiation. | Chronological changes of chromosome aberration rates related to accumulated doses in chronically exposed humans and animals at a low-dose-rate have not been well studied. C3H female specific pathogen-free mice (8 weeks of age) were chronically irradiated. Chromosome aberration rate in mouse splenocytes after long-term exposure to low-dose-rate (LDR) gamma-rays was serially determined by conventional Giemsa method. Incidence of dicentrics and centric rings increased almost linearly up to 8000 mGy following irradiation for about 400 days at a LDR of 20 mGy/day. Clear dose-rate effects were observed in the chromosome aberration frequencies between dose rates of 20 mGy/day and 200 Gy/day. Furthermore, the frequencies of complex aberrations increased as accumulated doses increased in LDR irradiation. This trend was also observed for the incidences of micronuclei and trisomies of chromosomes 5, 13 and 18 in splenocytes, detected by micronucleus assay and metaphase fluorescence in situ hybridization (FISH) method, respectively. Incidences of 2-4 micronuclei and trisomy increased in mouse splenocytes after irradiation of 8000 mGy at a LDR of 20 mGy/day. These complex chromosome aberrations and numerical chromosome aberrations seem to be induced indirectly after radiation exposure and thus the results indicate that continuous gamma-ray irradiation for 400 days at LDR of 20 mGy/day induced chromosomal instability in mice. These results are important to evaluate the biological effects of long-term exposure to LDR radiation in humans. | 18,930,838 |
Glu88 in the non-catalytic domain of acylpeptide hydrolase plays dual roles: charge neutralization for enzymatic activity and formation of salt bridge for thermodynamic stability. | Acylpeptide hydrolase of Aeropyrum pernix K1 is composed of a catalytic alpha/beta hydrolase domain and a non-catalytic beta-propeller domain. The Glu88 residue of the propeller domain is highly conserved in the prolyl oligopeptidase family and forms an inter-domain salt bridge with Arg526, a key residue for substrate binding. We have dissected the functions of Glu88 using site-directed mutagenesis, steady-state kinetics analyses, and molecular dynamics simulations. In E88A and E88A/R526K mutants, with a broken inter-domain salt bridge and a positive charge at position 526, catalytic activities for both a peptidase substrate and an esterase substrate were almost abolished. Analysis of the pH dependence of the mutants' reaction kinetics indicates that these mutations lead to changes in the electrostatic environment of the active site, which can be modulated by chloride ions. These findings indicate that the neutralization at position 526 is favorable for the activity of the enzyme, which is also verified by the catalytic behavior of E88A/R526V mutant. All mutants have lower thermodynamic stability than the wild-type. Therefore, Glu88 plays two major roles in the function of the enzyme: neutralizing the positive charge of Arg526, thereby increasing the enzymatic activity, and forming the Glu88-Arg526 salt bridge, thereby stabilizing the protein. | 18,930,847 |
Simultaneous analysis of the three hormones involved in spermatogenesis and their interrelation ratios. | The objectives of this non-interventional descriptive study was to determine the variation in serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone along with their ratios, among men having abnormal semen and their possible etiological role in male infertility. The study was carried out in the Reproductive Physiology Department of Public Health Laboratories Division, and National Institute of health (NIH), Islamabad during January 2004 to December 2005. Two hundred fifty married men who had presented with a complaint of infertility and who had an abnormal seminal profile on the basis of their prior semen analysis were included in the study. Subjects were classified as azoospermic (50), oligozoospermic (75), asthenozoospermic (50) and normozoospermic (75). In addition (50) normal male subjects, who were known to have fathered children, were included as controls. LH, FSH and testosterone levels were determined in serum by using enzyme immunoassay (EIA), using state-of-art Elecsys-2010 fully automatic immunology analyzer by Roche Diagnostics (USA). The FSH and LH level indicated inverse/negative correlation to sperm concentration in semen, while decreased testosterone levels were associated with depleted sperm concentration. The findings indicate that not only the altered/disturbed concentrations of gonadotrophins and androgenic hormones are responsible for male sub-fertility but also the disturbances in gonadotrophic: androgenic hormones ratios lead to infertility since these all hormones act synergistically. | 18,930,853 |
Development and evaluation of the Sinomenine transdermal patch. | Sinomenine transdermal patch was prepared and its properties were studied. The patches were produced by salivation method. The releasing rate in vitro of the patch was determined by HPLC. Peel test was used to evaluate the adhesion. Acute skin irritation test was performed in comparison with formalin (0.8%) by using mouse model. The Sinomenine TDDS Patch was prepared. The releasing rate in vitro followed the Higuchi equation (r>0.99), the releasing amount was beyond 90% in 24h. The peel adhesion to steel (N/25 mm) is 10 or above. The skin irritation tests showed negligible erythema and edema. The Sinomenine transdermal patch was prepared successfully and it may be beneficial for topical use. | 18,930,863 |
Inflammation and oxidative stress in vertebrate host-parasite systems. | Innate, inflammation-based immunity is the first line of vertebrate defence against micro-organisms. Inflammation relies on a number of cellular and molecular effectors that can strike invading pathogens very shortly after the encounter between inflammatory cells and the intruder, but in a non-specific way. Owing to this non-specific response, inflammation can generate substantial costs for the host if the inflammatory response, and the associated oxygen-based damage, get out of control. This imposes strong selection pressure that acts to optimize two key features of the inflammatory response: the timing of activation and resolution (the process of downregulation of the response). In this paper, we review the benefits and costs of inflammation-driven immunity. Our aim is to emphasize the importance of resolution of inflammation as a way of maintaining homeostasis against oxidative stress and to prevent the 'horror autotoxicus' of chronic inflammation. Nevertheless, host immune regulation also opens the way to pathogens to subvert host defences. Therefore, quantifying inflammatory costs requires assessing (i) short-term negative effects, (ii) delayed inflammation-driven diseases, and (iii) parasitic strategies to subvert inflammation. | 18,930,878 |
[Mechanisms for miniature dwarf characteristics of Mi-cro-Tom tomato and its application in plant functional genomics studies.]. | Micro-Tom is a miniature dwarf tomato, which can grow at a high density, has a short life cycle, and can be transformed efficiently. As a result, it became a new model plant for functional genomics study. The origin and biological characteristics of Micro-Tom were summarized. Recent advances in the mechanisms involved in the miniature dwarf trait, as well as the application of Micro-Tom in plant functional genomics study and the improved genetic transformation systems were reviewed. | 18,930,884 |
[Correlation analysis between MC3R and MC4R gene polymorphism and growth traits in pigeon.]. | The SNPs in partial coding sequence of MC3R and MC4R genes were identified by polymerase chain reaction followed by single strand conformation polymorphism (PCR-SSCP) analysis and DNA sequencing in shack-Kee and Columba domestica from Harbin area. Correlation analysis between MC3R and MC4R polymorphism and growth and body composition traits was carried by the least square analysis. The genotypes of T91G mutation in MC3R gene and A903G mutation in MC4R gene proved to have significant association with body weight, carcass weight, and holo-carcass weight in shack-Kee (P<0.05). The interaction of MC3R-T91G and MC4R-A903G was discussed through combination genotype analysis. The least square analysis showed that the combined genotype had significant association with holo-carcass weight (P<0.05). Multiple comparisons revealed that BBAA genotype birds had a higher holo-carcass weight than AABB genotype birds and BBAA genotype was the beneficial genotype for the growth of body weight. | 18,930,895 |
[Site-directed mutagenesis and function analysis of glgC gene from Escherichia coli.]. | Using genomic DNA of Escherichia coli JM109 as a template, glgC gene was amplified by polymerase chain reaction (PCR). The full coding sequence of this gene is 1296 bp. To get 3 mutants that amino acids changed: P295S (V121A, M151I, V334D), G336D and P295S/G336D (K109R) by recombinant PCR, respectively named 295+3, 336 and 295/336+1. The 3 mutants and the original glgC were subcloned into the prokaryotic expression vector pET-32a, and these recombinant expression plasmids were transformed into E. coli BL21 (DE3) for effective expression. The host cells were induced with IPTG and then identified by SDS-PAGE. A specific fused-expression product 67 kDa was detected, which was the same as the deduced protein. In the host cells above, the biological activities of the expressed products were detected by iodine vapor staining and glycogen content testing. The host cell transformed with the mutated gene-336 had higher glycogen content, which was identical to the gene-295/336+1. This confirmed that Pro295Ser could not reinforce the decrease of the feedback inhibition effect of the AGPase. Meanwhile, another host cell transformed with the mutated gene-295+3 showed decreased glycogen rather than the expected increasing glycogen. This might be caused by another mutation, Val334Asp in gene-295+3, which might induce the change of the allosteric region of the objective protein. | 18,930,900 |
Homo sapiens lactate dehydrogenase c (Ldhc) gene expression in cancer cells is regulated by transcription factor Sp1, CREB, and CpG island methylation. | The human testis-specific lactate dehydrogenase c gene (hLdhc) is transcribed only in cells of the germinal epithelium. Recently hLdhc was reported to express in a broad spectrum of tumors with relatively high frequency in lung cancer, melanoma, and breast cancer, and in some prostate cancers. Two melanoma cell lines that express the hLdhc gene, A375M and C81-61, were identified and were used to characterize the hLdhc promoter and explore transcriptional regulation of this gene. A 110-bp core promoter, including a conserved GC box and cyclic adenosine monophosphate-responsive element (CRE), were identified as essential for basal promoter activity. The methylation status of the CpG island (CGI) in the hLdhc core promoter sequence was analyzed in hLdhc-expressing and nonexpressing cells and human prostate tumor tissues. The CGI in 2 cell lines expressing the gene was hypomethylated whereas the DNA from cells that did not express hLdhc was hypermethylated. The role of methylation in regulating this promoter was confirmed by experimental induction of hLdhc transcription with the methylation inhibitor 5'aza-deoxycytidine. Quantitative analyses of the methylation level in the CGI were performed in prostate tumor tissues by pyrosequencing. Overall, these experiments demonstrated that hLdhc expression in cancer cells was regulated by transcription factor Sp1 and CREB and promoter CGI methylation. In addition, these findings suggest the possibility of developing a biomarker for cancer diagnosis/prognosis based on DNA methylation of the Ldhc gene. | 18,930,904 |
When "Simon says" doesn't work: alternatives to imitation for facilitating early speech development. | To provide clinicians with evidence-based strategies to facilitate early speech development in young children who are not readily imitating sounds. Relevant populations may include, but are not limited to, children with autism spectrum disorders, childhood apraxia of speech, and late-talking toddlers. Through multifaceted search procedures, we found experimental support for 6 treatment strategies that have been used to facilitate speech development in young children with developmental disabilities. Each strategy is highlighted within this article through a summary of the underlying rationale(s), empirical support, and specific examples of how it could be applied within intervention. Given the relatively sparse experimental data focused on facilitating speech in children who do not readily imitate, theoretical support emerges as particularly key and underscores the need for clinicians to consider why they are doing what they are doing. In addition, this review emphasizes the need for the research community to bridge the gap between pressing clinical needs and the limited evidence base that is currently available. | 18,930,909 |
Defining and measuring dysphagia following stroke. | To continue the development of a quantified, standard method to differentiate individuals with stroke and dysphagia from individuals without dysphagia. Videofluoroscopic swallowing studies (VFSS) were completed on a group of participants with acute stroke (n = 42) and healthy age-matched individuals (n = 25). Calibrated liquid volumes of 3, 5, 10, and 20 ml were administered during the VFSS. Six measures in 3 domains of bolus flow (timing, direction, and clearance) were measured. Values of these measures obtained from the control group were used to classify dysphagia within the participants. The use of a single measure or single liquid volumes to classify dysphagia did not distinguish between healthy adults and individuals following stroke with and without dysphagia. Abnormality on more than 1 measure across multiple volumes appears to be a more robust method in defining dysphagia for liquids. Our findings indicate that the definition of dysphagia is critical in determining whether persons are classified with disordered swallowing. The definition is dependent on materials and measures evaluated. Each measure provides independent aspects to the evaluation. Determining the level of importance of each depends on the purpose of the evaluation. | 18,930,911 |
The impact of embolic protection device and stent design on the outcome of CAS. | The importance of angioplasty and stenting in the treatment of carotid artery disease cannot be underestimated. Successful carotid stenting does not only depend on the operator's skills and experience but also on an adequate selection of cerebral protection devices, and carotid stents can help avoid neurological complications. A broad spectrum of carotid devices is currently on the market, and because each of them has its own advantages and disadvantages, it is virtually impossible to claim that one specific device is the best. The individual characteristics of each specific protection system or stent may make it an attractive choice in one circumstance but render it a less desirable option in other situations. The applicability depends primarily on the arterial anatomy and the specific details of the lesion being treated. But certainly, personal preferences and familiarity with a specific device may legitimately influence the decision to choose one over another. | 18,930,936 |
Hemodynamic changes associated with carotid artery interventions. | Carotid artery interventions can be associated with adverse hemodynamic changes, including bradycardia and hypotension. These hemodynamic changes are believed to be caused by direct stimulation of the carotid sinus baroreceptors, mimicking normal physiological response to rises in blood pressure. During open carotid surgery, these hemodynamic changes can be controlled by direct injection of medications that block fast voltage gated sodium channels in the neuron cell membrane, thus preventing depolarization of the presynaptic neuron in the carotid sinus. This form of control is difficult or impossible during percutaneous carotid interventions because direct access to the carotid artery and carotid sinus is not available. This discussion focuses on the cause, effects, and possible treatments for the hemodynamic changes associated with carotid artery stenting procedures. | 18,930,940 |
Tyrosines of human and mouse transferrin covalently labeled by organophosphorus agents: a new motif for binding to proteins that have no active site serine. | The expectation from the literature is that organophosphorus (OP) agents bind to proteins that have an active site serine. However, transferrin, a protein with no active site serine, was covalently modified in vitro by 0.5mM 10-fluoroethoxyphosphinyl-N-biotinamido pentyldecanamide, chlorpyrifos oxon, diisopropylfluorophosphate, dichlorvos, sarin, and soman. The site of covalent attachment was identified by analyzing tryptic peptides in the mass spectrometer. Tyr 238 and Tyr 574 in human transferrin and Tyr 238, Tyr 319, Tyr 429, Tyr 491, and Tyr 518 in mouse transferrin were labeled by OP. Tyrosine in the small synthetic peptide ArgTyrThrArg made a covalent bond with diisopropylfluorophosphate, chlorpyrifos oxon, and dichlorvos at pH 8.3. These results, together with our previous demonstration that albumin and tubulin bind OP on tyrosine, lead to the conclusion that OP bind covalently to tyrosine, and that OP binding to tyrosine is a new OP-binding residue. The OP-reactive tyrosines are activated by interaction with Arg or Lys. It is suggested that many proteins in addition to those already identified may be modified by OP on tyrosine. The extent to which tyrosine modification by OP can occur in vivo and the toxicological implications of such modifications require further investigation. | 18,930,948 |
Immunohistochemical evaluation of ProEx C in human papillomavirus-induced lesions of the cervix. | Considering the sparse information about the clinical utility of the novel immunohistochemical marker ProEx C in histological sections, a decision was taken to study the pattern of ProEx C expression in normal/benign cervical epithelium (N/B), low-grade squamous intraepithelial lesion (LGSIL) and high-grade squamous intraepithelial lesion (HGSIL), as well as the association of ProEx C expression with human papillomavirus (HPV) genotypes. 100 cervical samples, including 21 N/B cervices, 16 LGSILs, 61 HGSILs and two cervical invasive carcinomas, were obtained from conisation and hysterectomy. Surgical specimens were arranged in three tissue microarrays and stained for ProEx C. Ninety-three samples were HPV genotyped. Genotyping was performed by DNA amplification and hybridisation with genotype-specific probes on a low-density DNA array. ProEx C-positive expression in more than the lower third of the epithelium was observed in 14.3% of N/B, 62.5% of LGSIL and 90.2% of HGSIL. Seventy percent of HPV positivity was found in cases with expression in more than the lower third of the epithelium. Of 31 cases that were positive for HPV16, 16.1% showed ProEx C expression restricted to one or two basal layers, and 83.9% showed ProEx C expression in more than the lower third of the epithelium. ProEx C is significantly associated with HPV16 infection and is a useful adjunct in the identification of LGSIL and HGSIL in histological sections when expressed in more than the lower third of the epithelium. | 18,930,984 |
Effect of CHEK2 missense variant I157T on the risk of breast cancer in carriers of other CHEK2 or BRCA1 mutations. | Carriers of heterozygous mutations in CHEK2 or BRCA1 are at increased risk of breast cancer. These mutations are rare and a very small number of women in a population will carry two mutations. However, it is of interest to estimate the breast cancer risks associated with carrying two mutations because this information may be informative for genetic counsellors and may provide clues to the carcinogenic process. We genotyped 7782 Polish breast cancer patients and 6233 controls for seven founder mutations in BRCA1 and CHEK2. Odds ratios (OR) and 95% confidence intervals (CI) were estimated for the mutations, singly and in combination. Of the 7782 women with breast cancer, 1091 had one mutation (14.0%) and 37 had two mutations (0.5%). Compared to controls, the odds ratio for a BRCA1 mutation in isolation was 13.1 (95% CI 8.2 to 21). The odds ratio was smaller for BRCA1 mutation carriers who also carried a CHEK2 mutation (OR 6.6, 95% CI 1.5 to 29), but the difference was not statistically significant. In contrast, the odds ratio for women who carried two CHEK2 mutations (OR 3.9, 95% CI 1.5 to 10) was greater than that for women who carried one CHEK2 mutation (OR 1.9, 95% CI 1.6 to 2.1). The odds ratio for women who carried both a truncating mutation and the missense mutation in CHEK2 was 7.0 (95% CI 0.9 to 56) and was greater than for women who carried the truncating mutation alone (OR 3.3, 95% CI 2.4 to 4.3) or the missense mutation alone (OR 1.6, 95% CI 1.4 to 1.9), but the difference was not statistically significant. Our study suggests that the risk of breast cancer in carriers of a deleterious CHEK2 mutation is increased if the second allele is the I157T missense variant. However, the presence of a CHEK2 mutation in women with a BRCA1 mutation may not increase their risk beyond that of the BRCA1 mutation alone. These suggestive findings need to be verified in other studies. | 18,930,998 |
Cerebrospinal fluid anti-myelin antibodies are related to magnetic resonance measures of disease activity in multiple sclerosis. | Recent studies reported contrasting results with respect to the presence of anti-myelin protein antibodies in multiple sclerosis (MS) and their relation with disease activity. This may be due to the heterogeneous specificity of autoantibodies in MS and the inability of most methods to detect pathogenically relevant antibodies. Here, myelin particles were used to detect anti-myelin antibodies in the CSF of MS patients. Subsequently, their relation with MRI parameters was evaluated. Anti-myelin IgG antibody reactivity was determined in the CSF of patients with MS (n = 65) and clinically isolated syndrome (CIS, n = 37) using a novel flow cytometry based assay. In addition, the CSF of patients with other neurological diseases (OND, n = 17), inflammatory neurological diseases (IND, n = 33) and controls (n = 22) was tested. Compared with controls, increased anti-myelin IgG antibody reactivity was most frequently found in the CSF of patients with CIS (46%, p = 0.002), relapsing-remitting MS (56%, p<0.001) and secondary progressive MS (55%, p<0.001), together constituting 85% of all positive CSF samples. In contrast, elevated anti-myelin IgG antibody reactivity was present in a minority of IND patients (21%), marginally present in controls (5%) and absent in OND patients (0%). Most strikingly, anti-myelin IgG antibody reactivity was related to the number of T2 lesions (r = 0.31, p = 0.041) and gadolinium enhancing T1 lesions (r = 0.37, p = 0.016) on brain MRI in CIS and relapse onset MS patients. CSF anti-myelin IgG antibodies are promising specific biomarkers in CIS and relapse onset MS and correlate with MR measures of disease activity. | 18,931,010 |
Central and peripheral integration of interrenal and thyroid axes signals in common carp (Cyprinus carpio L.). | In teleostean fishes the hypothalamic-pituitary-thyroid axis (HPT axis) and the hypothalamic-pituitary-interrenal axis (HPI axis) regulate the release of thyroid hormones (THs) and cortisol respectively. Since many actions of both hormones are involved in the regulation of metabolic processes, communication between both signal pathways can be anticipated. In this study, we describe central and peripheral sites for direct interaction between mediators of both neuroendocrine axes in the common carp (Cyprinus carpio). Despite suggestions in the literature that CRH is thyrotropic in some fish; we were not able to establish stimulatory effects of CRH on the expression of the pituitary TSHbeta subunit gene. In preoptic area tissue incubated with 10(-7) M thyroxine (T(4)) a 2 x 9-fold increase in the expression of CRH-binding protein (CRHBP) was observed. Thus, T(4) could reduce the bioavailable hypothalamic crh via the up regulation of crhbp expression and hence down regulate the HPI axis. At the peripheral level, cortisol (10(-6) M), ACTH (10(-7) M), and alpha-MSH (10(-7) M) stimulate the release of T(4) from kidney and head kidney fragments, which contain all functional thyroid follicles in carp, by two- to fourfold. The substantiation of three pituitary thyrotropic factors, viz. TSH, ACTH, and alpha-MSH, in common carp, allows for an integration of central thyrotropic signals. Clearly, two sites for interaction between the HPT axis, the HPI axis, and alpha-MSH are present in common carp. These interactions may be key to the proper regulation of general metabolism in this fish. | 18,931,024 |
Sympathetic neurogenic Ca2+ signalling in rat arteries: ATP, noradrenaline and neuropeptide Y. | The sympathetic nervous system (SNS) plays an essential role in the control of total peripheral vascular resistance by controlling the contraction of small arteries. The SNS also exerts long-term trophic influences in health and disease; SNS hyperactivity accompanies most forms of human essential hypertension, obesity and heart failure. At their junctions with smooth muscle cells, the peri-arterial sympathetic nerves release ATP, noradrenaline (NA) and neuropeptide Y (NPY) onto smooth muscle cells. Confocal Ca(2+) imaging studies reveal that ATP and NA each produce unique types of postjunctional Ca(2+) signals and consequent smooth muscle cell contractions. Neurally released ATP activates postjunctional P2X(1) receptors to produce local, non-propagating Ca(2+) transients, termed 'junctional Ca(2+) transients', or 'jCaTs'. Neurally released NA binds to alpha(1)-adrenoceptors and can activate Ca(2+) waves or more uniform global changes in [Ca(2+)]. Neurally released NPY does not appear to produce Ca(2+) transients directly, but significantly modulates NA-induced Ca(2+) signalling. The neural release of ATP and NA, as judged by postjunctional Ca(2+) signals, electrical recording of excitatory junction potentials and carbon fibre amperometry to measure NA, varies markedly with the pattern of nerve activity. This probably reflects both pre- and postjunctional mechanisms, which are not yet fully understood. These phenomena, together with different temporal patterns of sympathetic nerve activity in different regional circulations, are probably an important mechanistic basis of the important selective regulation of regional vascular resistance and blood flow by the sympathetic nervous system. | 18,931,047 |
An avian live attenuated master backbone for potential use in epidemic and pandemic influenza vaccines. | The unprecedented emergence in Asia of multiple avian influenza virus (AIV) subtypes with a broad host range poses a major challenge in the design of vaccination strategies that are both effective and available in a timely manner. The present study focused on the protective effects of a genetically modified AIV as a source for the preparation of vaccines for epidemic and pandemic influenza. It has previously been demonstrated that a live attenuated AIV based on the internal backbone of influenza A/Guinea fowl/Hong Kong/WF10/99 (H9N2), called WF10att, is effective at protecting poultry species against low- and high-pathogenicity influenza strains. More importantly, this live attenuated virus provided effective protection when administered in ovo. In order to characterize the WF10att backbone further for use in epidemic and pandemic influenza vaccines, this study evaluated its protective effects in mice. Intranasal inoculation of modified attenuated viruses in mice provided adequate protective immunity against homologous lethal challenges with both the wild-type influenza A/WSN/33 (H1N1) and A/Vietnam/1203/04 (H5N1) viruses. Adequate heterotypic immunity was also observed in mice vaccinated with modified attenuated viruses carrying H7N2 surface proteins. The results presented in this report suggest that the internal genes of a genetically modified AIV confer similar protection in a mouse model and thus could be used as a master donor strain for the generation of live attenuated vaccines for epidemic and pandemic influenza. | 18,931,063 |
Stimulation of interleukin-13 expression by human T-cell leukemia virus type 1 oncoprotein Tax via a dually active promoter element responsive to NF-kappaB and NFAT. | The human T-cell leukemia virus type 1 (HTLV-1) Tax oncoprotein transforms human lymphocytes and is critical for the pathogenesis of HTLV-1-induced adult T-cell leukaemia. In HTLV-transformed cells, Tax upregulates interleukin (IL)-13, a cytokine with proliferative and anti-apoptotic functions that is linked to leukaemogenesis. Tax-stimulated IL-13 is thought to result in autocrine stimulation of HTLV-infected cells and thus may be relevant to their growth. The causal transactivation of the IL-13 promoter by Tax is predominantly dependent on a nuclear factor of activated T cells (NFAT)-binding P element. Here, it was shown that the isolated IL-13 Tax-responsive element (IL13TaxRE) was sufficient to mediate IL-13 transactivation by Tax and NFAT1. However, cyclosporin A, a specific NFAT inhibitor, revealed that Tax transactivation of IL13TaxRE or wild-type IL-13 promoter was independent of NFAT and that NFAT did not contribute to IL-13 upregulation in HTLV-transformed cells. By contrast, Tax stimulation was repressible by an efficient nuclear factor (NF)-kappaB inhibitor (IkBaDN), indicating the requirement for NF-kappaB. The capacity of NF-kappaB to stimulate IL13TaxRE was demonstrated by a strong response to NF-kappaB in reporter assays and by direct binding of NF-kappaB to IL13TaxRE. Thus, IL13TaxRE in the IL-13 promoter represents a dually active promoter element responsive to NF-kappaB and NFAT. Together, these results indicate that Tax causes IL-13 upregulation in HTLV-1-infected cells via NF-kappaB. | 18,931,077 |
Rice dwarf virus is engulfed into and released via vesicular compartments in cultured insect vector cells. | Vector insect cells infected with Rice dwarf virus had vesicular compartments containing viral particles located adjacent to the viroplasm when examined by transmission electron and confocal microscopy. Such compartments were often at the periphery of infected cells. Inhibitors of vesicular transport, brefeldin A and monensin, and an inhibitor of myosin motor activity, butanedione monoxime, abolished the formation of such vesicles and prevented the release of viral particles from infected cells without significant effects on virus multiplication. Furthermore, the actin-depolymerizing drug, cytochalasin D, inhibited the formation of actin filaments without significantly interfering with formation of vesicular compartments and the release of viruses from treated cells. These results together revealed intracellular vesicular compartments as a mode for viral transport in and release from insect vector cells infected with a plant-infecting reovirus. | 18,931,091 |
Moraxella catarrhalis M35 is a general porin that is important for growth under nutrient-limiting conditions and in the nasopharynges of mice. | Moraxella catarrhalis is a gram-negative respiratory pathogen that is an important causative agent for otitis media and exacerbations of chronic obstructive pulmonary disease. We have previously predicted the outer membrane protein M35 to be a general porin, and in the current study, we have investigated the function of M35 and its importance for survival of M. catarrhalis in vivo. Lipid bilayer experiments reveal that refolded M35 functions as a channel that is typical of gram-negative bacterial porins. M35 forms wide and water-filled channels with a single-channel conductance of about 1.25 nS in 1 M KCl solution and has only a small selectivity for cations over anions. When the in vitro growth characteristics of two M35 deletion mutant strains of M. catarrhalis were compared to the wild-type parent isolates, the growth of the mutant strains was inhibited only under nutrient-poor conditions. This growth defect could be eliminated by additional glutamic acid, but not additional aspartic acid, glycine, sucrose, or glucose. The mutant strains compensated for the lack of M35 by enhancing their uptake of glutamic acid, and this enhanced rate of glutamic acid uptake was attributed to the compensatory upregulation of a protein of approximately 40 kDa. M35 was also found to be essential for nasal colonization of mice, demonstrating that its presence is essential for survival of M. catarrhalis in vivo. These results suggest that M35 is a general porin that is necessary for the uptake of important energy sources by M. catarrhalis and that it is likely that M35 is an essential functional protein for in vivo colonization. | 18,931,134 |
The chloroplast DnaJ homolog CDJ1 of Chlamydomonas reinhardtii is part of a multichaperone complex containing HSP70B, CGE1, and HSP90C. | We report on the molecular and biochemical characterization of CDJ1, one of three zinc-finger-containing J-domain proteins encoded by the Chlamydomonas reinhardtii genome. Fractionation experiments indicate that CDJ1 is a plastidic protein. In the chloroplast, CDJ1 was localized to the soluble stroma fraction, but also to thylakoids and to low density membranes. Although the CDJ1 gene was strongly heat shock inducible, CDJ1 protein levels increased only slightly during heat shock. Cellular CDJ1 concentrations were close to those of heat shock protein 70B (HSP70B), the major HSP70 in the Chlamydomonas chloroplast. CDJ1 complemented the temperature-sensitive phenotype of an Escherichia coli mutant lacking its dnaJ gene and interacted with E. coli DnaK, hence classifying it as a bona fide DnaJ protein. In soluble cell extracts, CDJ1 was found to organize into stable dimers and into complexes of high molecular mass. Immunoprecipitation experiments revealed that CDJ1 forms common complexes with plastidic HSP90C, HSP70B, and CGE1. In blue native-polyacrylamide gel electrophoresis, all four (co)chaperones migrated at 40% to 90% higher apparent than calculated molecular masses, indicating that greatest care must be taken when molecular masses of protein complexes are estimated from their migration relative to standard native marker proteins. Immunoprecipitation experiments from size-fractioned soluble cell extracts suggested that HSP90C and HSP70B exist as preformed complex that is joined by CDJ1. In summary, CDJ1 and CGE1 are novel cohort proteins of the chloroplast HSP90-HSP70 multichaperone complex. As HSP70B, CDJ1, and CGE1 are derived from the endosymbiont, whereas HSP90C is of eukaryotic origin, we observe in the chloroplast the interaction of two chaperone systems of distinct evolutionary origin. | 18,931,144 |
Effect of enzyme addition on the nutritive value of high oleic acid sunflower seeds in chicken diets. | Two experiments were conducted to evaluate the effects of enzyme addition in chicken diets containing high oleic acid sunflower seeds (HOASS). In the first experiment (4 to 21 d of age), enzyme addition (lipase, phospholipase, and a combination of these) was used at the inclusion level of 1 g/kg in diets containing HOASS (250 g/kg) compared with a control corn-soybean diet. Weight gain, feed consumption, relative liver weight, fat digestibility, and amylase, lipase, serum lactate dehydrogenase (LDH), and creatine phosphokinase (CPK) activities were reduced, and feed conversion, relative duodenum, jejunum, ileum, and ceca lengths, plasma uric acid, cholesterol, and glucose concentrations were increased in the unsupplemented HOASS diet compared with the control diet. The addition of enzymes to the HOASS diet increased weight gain, feed consumption, relative pancreas and liver weights, fat digestibility, amylase and lipase activities, plasma uric acid, calcium, serum LDH and CPK, and total protein concentration and reduced feed conversion, relative spleen weight, relative duodenum, jejunum, ileum, and ceca lengths, plasma cholesterol, and glucose compared with the unsupplemented HOASS diet. In the second experiment (0 to 21 d of age), the same enzymes (0.5 g/kg each) were included in diets containing 150 g/kg of HOASS compared with a conventional sunflower meal diet (150 g/kg). The HOASS diet did not affect performance but reduced relative pancreas and abdominal fat weights and relative duodenum and ceca lengths, and increased crude fat, CP, and essential and nonessential amino acid digestibilities (except Ser, which was reduced) compared with the control diet. The addition of enzymes in the HOASS diet increased weight gain, feed consumption, and relative pancreas weight and reduced feed conversion, CP, and essential and nonessential amino acid digestibilities compared with the unsupplemented HOASS diet. In conclusion, the addition of 250 g of HOASS/kg in the diets caused a negative effect on performance, digestive organ sizes, fat and protein digestibilities, and pancreatic enzymes and modified blood parameters. However, the inclusion of HOASS at 150 g/kg improved some of these parameters and amino acid digestibilities. The enzyme addition counteracted some of these effects. | 18,931,181 |
Improved neuromuscular blockade using a novel neuromuscular blockade advisory system: a randomized, controlled, clinical trial. | Conventional incremental bolus administration of neuromuscular blocking (NMB) drugs is associated with limitations in intraoperative control, potential delays in recovery, and residual blockade in the postanesthetic period. To overcome such limitations, we developed a novel adaptive control computer program, the Neuromuscular Blockade Advisory System (NMBAS). The NMBAS advises the anesthesiologist on the timing and dose of NMB drugs based on a sixth-order Laguerre model and the history of the patient's electromyographic responses. Here, we tested the hypothesis that the use of the NMBAS improves NMB compared to standard care. We conducted a prospective, randomized, controlled, blinded, parallel-group, clinical trial with n = 73 patients (ASA physical status I-III) undergoing abdominal surgery under general anesthesia > or =1.5 h with NMB using rocuronium. Patients were allocated to standard care or NMBAS-guided rocuronium administration. The primary outcome variable was the incidence of intraoperative events reflecting inadequate NMB. Secondary outcome variables included train-of-four (TOF) ratios at the end of surgery before reversal, the total doses of rocuronium, reversal agents, anesthetics and other drugs, the incidence of postoperative adverse events, and the incidence of anesthesiologist noncompliance with NMBAS recommendations. Of 73 enrolled patients, n = 30 per group were eligible for analysis. Patient demographics were comparable between the groups. The incidence in total intraoperative events associated with inadequate NMB was significantly lower in the NMBAS group compared to standard care (8/30 vs 19/30; P = 0.004). Mean TOF ratios at the end of surgery before reversal were higher in the NMBAS group (0.59 [95% CI, 0.48-0.69] vs 0.14 [95% CI, 0.04-0.24]; P < 0.0001). Total administered doses of rocuronium, reversal drugs, and other drugs, and the incidence of postoperative adverse events were not different. Compared to standard practice, NMBAS-guided care was associated with improved NMB quality and higher TOF ratios at the end of surgery, potentially reducing the risk of residual NMB and improving perioperative patient safety. | 18,931,219 |
Intravascular injection during ultrasound-guided axillary block: negative aspiration can be misleading. | Needle visualization is an advantage of ultrasound-guided nerve blocks compared to traditional methods of nerve localization. However, visualization of local anesthetic spread is also important. During an ultrasound-guided axillary block, there was negative aspiration for blood. However, the absence of local anesthetic spread on ultrasound imaging suggested possible intravascular injection. The ultrasound transducer was removed from the patient's arm and venous blood was aspirated from the nerve block needle. Pressure applied to an ultrasound transducer can occlude venous structures making negative aspiration of blood unreliable for excluding intravascular needle placement. | 18,931,244 |
Small- and large-scale conformational changes of adenylate kinase: a molecular dynamics study of the subdomain motion and mechanics. | Adenylate kinase, an enzyme that catalyzes the phosphoryl transfer between ATP and AMP, can interconvert between the open and catalytically potent (closed) forms even without binding ligands. Several aspects of the enzyme elasticity and internal dynamics are analyzed here by atomistic molecular dynamics simulations covering a total time span of 100 ns. This duration is sufficiently long to reveal a partial conversion of the enzyme that proceeds through jumps between structurally different substates. The intra- and intersubstates contributions to the enzyme's structural fluctuations are analyzed and compared both in magnitude and directionality. It is found that, despite the structural heterogeneity of the visited conformers, the generalized directions accounting for conformational fluctuations within and across the substates are mutually consistent and can be described by a limited set of collective modes. The functional-oriented nature of the consensus modes is suggested by their good overlap with the deformation vector bridging the open and closed crystal structures. The consistency of adenylate kinase's internal dynamics over timescales wide enough to capture intra- and intersubstates fluctuations adds elements in favor of the recent proposal that the free (apo) enzyme possesses an innate ability to sustain the open/close conformational changes. | 18,931,260 |
Thought processes associated with reluctance in gay men to be tested for HIV. | This study explored the thought processes that are associated with reluctance in gay men to be tested for HIV antibodies. The sample comprised 97 men who had not been tested for at least four years; 69 had never been tested. They were asked to imagine that someone had suggested that they be tested very soon and to identify, from the list provided, any negative thoughts prompted by this suggestion. The most commonly reported thoughts were that testing was unnecessary because risks had not been taken, that it was unnecessary because there were no symptoms, and that there was no urgency to be tested. Data were explored by means of factor analysis and comparisons across subgroups differing in risk level. The results are interpreted as indicating the use of rationalizations to buttress a decision not to be tested, the powerful influence on HIV decision-making exerted by salient perceptible features, and the 'status quo bias'. Techniques that could be used to encourage testing in gay men are discussed. | 18,931,273 |
Influence of high pressure on the dimerization of ToxR, a protein involved in bacterial signal transduction. | High hydrostatic pressure (HHP) is suggested to influence the structure and function of membranes and/or integrated proteins. We demonstrate for the first time HHP-induced dimer dissociation of membrane proteins in vivo with Vibrio cholerae ToxR variants in Escherichia coli reporter strains carrying ctx::lacZ fusions. Dimerization ceased at 20 to 50 MPa depending on the nature of the transmembrane segments rather than on changes in the ToxR lipid bilayer environment. | 18,931,287 |
Multilocus genotyping assays for single nucleotide polymorphism-based subtyping of Listeria monocytogenes isolates. | Listeria monocytogenes is responsible for serious invasive illness associated with consumption of contaminated food and places a significant burden on public health and the agricultural economy. We recently developed a multilocus genotyping (MLGT) assay for high-throughput subtype determination of L. monocytogenes lineage I isolates based on interrogation of single nucleotide polymorphisms (SNPs) via multiplexed primer extension reactions. Here we report the development and validation of two additional MLGT assays that address the need for comprehensive DNA sequence-based subtyping of L. monocytogenes. The first of these novel MLGT assays targeted variation segregating within lineage II, while the second assay combined probes for lineage III strains with probes for strains representing a recently characterized fourth evolutionary lineage (IV) of L. monocytogenes. These assays were based on nucleotide variation identified in >3.8 Mb of comparative DNA sequence and consisted of 115 total probes that differentiated 93% of the 100 haplotypes defined by the multilocus sequence data. MLGT reproducibly typed the 173 isolates used in SNP discovery, and the 10,448 genotypes derived from MLGT analysis of these isolates were consistent with DNA sequence data. Application of the MLGT assays to assess subtype prevalence among isolates from ready-to-eat foods and food-processing facilities indicated a low frequency (6.3%) of epidemic clone subtypes and a substantial population of isolates (>30%) harboring mutations in inlA associated with attenuated virulence in cell culture and animal models. These mutations were restricted to serogroup 1/2 isolates, which may explain the overrepresentation of serotype 4b isolates in human listeriosis cases. | 18,931,295 |
Biomechanics of a convergently derived prey-processing mechanism in fishes: evidence from comparative tongue bite apparatus morphology and raking kinematics. | A tongue-bite apparatus (TBA) governs raking behaviors in two major and unrelated teleost lineages, the osteoglossomorph and salmoniform fishes. We present data on comparative morphology and kinematics from two representative species, the rainbow trout (Oncorhynchus mykiss) and the Australian arowana (Scleropages jardinii), which suggest that both the TBA and raking are convergently derived in these lineages. Similar TBA morphologies were present, except for differences in TBA dentition and shape of the novel cleithrobranchial ligament (CBL), which is arc-shaped in O. mykiss and straight in S. jardinii. Eight kinematic variables were used to quantify motion magnitude and maximum-timing in the kinematic input mechanisms of the TBA. Five variables differed inter-specifically (pectoral girdle retraction magnitude and timing, cranial and hyoid elevation and gape-distance timing), yet an incomplete taxon separation across multivariate kinematic space demonstrated an overall similarity in raking behavior. An outgroup analysis using bowfin (Amia calva) and pickerel (Esox americanus) to compare kinematics of raking with chewing and prey-capture provided robust quantitative evidence of raking being a convergently derived behavior. Support was also found for the notion that raking more likely evolved from the strike, a functionally distinct behavior, than from chewing, an alternative prey-processing behavior. Based on raking kinematic and muscle-activity data, we propose biomechanical models of the three input mechanisms that govern kinematics of the basihyal output mechanism during the raking power stroke: (1) cranial elevation protracts the upper TBA jaw from the lower (basihyal) TBA jaw; (2) basihyal retraction is caused directly by contraction of the sternohyoideus (SH); (3) hypaxial shortening, relayed via the pectoral girdle and SH-CBL complex, is an indirect basihyal retraction mechanism modeled as a four-bar linkage. These models will aid future analyses mapping structural and functional traits to the evolution of behaviors. | 18,931,311 |
To break a coralline: mechanical constraints on the size and survival of a wave-swept seaweed. | Previous studies have hypothesized that wave-induced drag forces may constrain the size of intertidal organisms by dislodging or breaking organisms that exceed some critical dimension. In this study, we explored constraints on the size of the articulated coralline alga Calliarthron, which thrives in wave-exposed intertidal habitats. Its ability to survive depends critically upon its segmented morphology (calcified segments separated by flexible joints or ;genicula'), which allows otherwise rigid fronds to bend and thereby reduce drag. However, bending also amplifies stress within genicula near the base of fronds. We quantified breakage of genicula in bending by applying known forces to fronds until they broke. Using a mathematical model, we demonstrate the mitigating effect of neighboring fronds on breakage and show that fronds growing within dense populations are no more likely to break in bending than in tension, suggesting that genicular morphology approaches an engineering optimum, possibly reflecting adaptation to hydrodynamic stress. We measured drag in a re-circulating water flume (0.23-3.6 m s(-1)) and a gravity-accelerated water flume, which generates jets of water that mimic the impact of breaking waves (6-10 m s(-1)). We used frond Reynolds number to extrapolate drag coefficients in the field and to predict water velocities necessary to break fronds of given sizes. Laboratory data successfully predicted frond sizes found in the field, suggesting that, although Calliarthron is well adapted to resist breakage, wave forces may ultimately limit the size of intertidal fronds. | 18,931,316 |
Metabolomic biomarkers in a model of asthma exacerbation: urine nuclear magnetic resonance. | Airway obstruction in patients with asthma is associated with airway dysfunction and inflammation. Objective measurements including sputum analysis can guide therapy, but this is often not possible in typical clinical settings. Metabolomics is the study of molecules generated by metabolic pathways. We hypothesize that airway dysfunction and inflammation in an animal model of asthma would produce unique patterns of urine metabolites measured by multivariate statistical analysis of high-resolution proton nuclear magnetic resonance ((1)H NMR) spectroscopy data. To develop a noninvasive means of monitoring asthma status by metabolomics and urine sampling. Five groups of guinea pigs were studied: control, control treated with dexamethasone, sensitized (ovalbumin, administered intraperitoneally), sensitized and challenged (ovalbumin, administered intraperitoneally, plus ovalbumin aerosol), and sensitized-challenged with dexamethasone. Airway hyperreactivity (AHR) to histamine (administered intravenously) and inflammation were measured. Multivariate statistical analysis of NMR spectra based on a library of known urine metabolites was performed by partial least-squares discriminant analysis. In addition, the raw NMR spectra exported as xy-trace data underwent linear discriminant analysis. Challenged guinea pigs developed AHR and increased inflammation compared with sensitized or control animals. Dexamethasone significantly improved AHR. Using concentration differences in metabolites, partial least-squares discriminant analysis could discriminate challenged animals with 90% accuracy. Using only three or four regions of the NMR spectra, linear discriminant analysis-based classification demonstrated 80-90% separation of the animal groups. Urine metabolites correlate with airway dysfunction in an asthma model. Urine NMR analysis is a promising, noninvasive technique for monitoring asthma in humans. | 18,931,331 |
Surrogate decision-makers' perspectives on discussing prognosis in the face of uncertainty. | Many physicians are reluctant to discuss a patient's prognosis when there is significant prognostic uncertainty. We sought to understand surrogate decision makers' views regarding whether physicians should discuss prognosis in the face of uncertainty. We conducted semi-structured interviews with 179 surrogates for 142 incapacitated patients at high risk of death in four intensive care units at an academic medical center. The interviews explored surrogates' attitudes about whether physicians should discuss prognosis when they cannot be certain their prognostic estimates are correct. We used constant comparative methods to analyze the transcripts. Validation methods included triangulation by multidisciplinary analysis and member checking. Eighty-seven percent (155/179) of surrogates wanted physicians to discuss an uncertain prognosis. We identified five main reasons for this, including surrogates' belief that prognostic uncertainty is unavoidable, that physicians are their only source for prognostic information, and that discussing prognostic uncertainty leaves room for realistic hope, increases surrogates' trust in the physician, and signals a need to prepare for possible bereavement. Twelve percent (22/179) of surrogates felt that discussions about an uncertain prognosis should be avoided. The main explanation was that it is not worth the potential emotional distress if the prognostications are incorrect. Surrogates suggested that physicians should explicitly discuss uncertainty when prognosticating. The majority of surrogates of patients that are critically ill want physicians to disclose their prognostic estimates even if they cannot be certain they are correct. This stems from surrogates' belief that prognostic uncertainty is simultaneously unavoidable and acceptable. | 18,931,332 |
Airflow limitation in smokers is associated with subclinical atherosclerosis. | Chronic obstructive pulmonary disease (COPD) is associated with increased morbidity and mortality from cardiovascular disease. Although a close association between COPD and atherosclerosis has been speculated, such scientific information is limited. To evaluate subclinical atherosclerosis in smokers with airflow limitation. The subjects of this study were healthy middle-aged men. Smokers with airflow limitation (n = 61) and age-matched control smokers (n = 122) and control never-smokers (n = 122) without airflow limitation were included in the present study. Subjects with diabetes, acute infection, and respiratory disease other than COPD were excluded beforehand. All subjects underwent chest radiogram, spirometry, blood sampling, and carotid ultrasonography. We determined carotid intima-media thickness and focal atheromatous plaque as indicators of subclinical atherosclerosis. Mean carotid intima-media thickness was greater in smokers with airflow limitation than in control smokers (P < 0.01) and control never-smokers (P < 0.005). Focal carotid plaque was significantly more prevalent in smokers with airflow limitation than in control never-smokers (P < 0.005). Multivariate analyses showed significant associations between thickened intima-media thickness and decreased percent predicted FEV(1) (P = 0.001) and between plaque and log(10) C-reactive protein (P = 0.013) independent of age, pack-years of smoking, body mass index, peripheral mean arterial pressure, heart rate, glucose, and low-density lipoprotein cholesterol. Smokers with airflow limitation had exaggerated subclinical atherosclerosis. This study suggests that middle-aged men who are susceptible to COPD may also be susceptible to vascular atherosclerosis by smoking, and atherosclerotic change starts early in the disease process of COPD. | 18,931,335 |
P3DB: a plant protein phosphorylation database. | P(3)DB (http://www.p3db.org/) provides a resource of protein phosphorylation data from multiple plants. The database was initially constructed with a dataset from oilseed rape, including 14,670 nonredundant phosphorylation sites from 6382 substrate proteins, representing the largest collection of plant phosphorylation data to date. Additional protein phosphorylation data are being deposited into this database from large-scale studies of Arabidopsis thaliana and soybean. Phosphorylation data from current literature are also being integrated into the P(3)DB. With a web-based user interface, the database is browsable, downloadable and searchable by protein accession number, description and sequence. A BLAST utility was integrated and a phosphopeptide BLAST browser was implemented to allow users to query the database for phosphopeptides similar to protein sequences of their interest. With the large-scale phosphorylation data and associated web-based tools, P(3)DB will be a valuable resource for both plant and nonplant biologists in the field of protein phosphorylation. | 18,931,372 |
The N-clasp of human DNA polymerase kappa promotes blockage or error-free bypass of adenine- or guanine-benzo[a]pyrenyl lesions. | DNA bypass polymerases are utilized to transit bulky DNA lesions during replication, but the process frequently causes mutations. The structural origins of mutagenic versus high fidelity replication in lesion bypass is therefore of fundamental interest. As model systems, we investigated the molecular basis of the experimentally observed essentially faithful bypass of the guanine 10S-(+)-trans-anti-benzo[a]pyrene-N(2)-dG adduct by the Y-family human DNA polymerase kappa, and the observed blockage of pol kappa produced by the adenine 10S-(+)-trans-anti-benzo[a]pyrene-N(2)-dA adduct. These lesions are derived from the most tumorigenic metabolite of the ubiquitous cancer-causing pollutant, benzo[a]pyrene. We compare our results for the dG adduct with our earlier studies for the pol kappa archaeal homolog Dpo4, which processes the same lesion in an error-prone manner. Molecular modeling, molecular mechanics calculations and molecular dynamics simulations were utilized. Our results show that the pol kappa N-clasp is a key structural feature that accounts for the dA adduct blockage and the near-error-free bypass of the dG lesion. Absence of the N-clasp in Dpo4 explains the error-prone processing of the same lesion by this enzyme. Thus, our studies elucidate structure-function relationships in the fidelity of lesion bypass. | 18,931,375 |
Low level IGF-1 and common variable immune deficiency: an unusual combination. | A relation between growth hormone (GH) deficiency and immunoglobulin deficiency has been suggested previously in a few cases. We describe a patient with an insulin-like growth factor 1 (IGF-1) deficiency and common variable immune deficiency and briefly review earlier publications on the possible interaction between IGF-1 and the immune system. IGF-1 is the downstream mediator of GH. In this patient, GH and IGF-1 levels were both low. The GH response to a GH-releasing hormone test was normal whereas no subsequent IGF-1 response was seen. In our cohort of 14 patients with hypogammaglobulinaemia, two turned out to have slightly decreased IGF-1 serum levels and one patient with a thymoma had an increased IGF-1 level. Even though IGF-1 may be connected to B lymphocyte differentiation, in this patient we hypothesise there is a common impairment in the IGF-1 and IgG pathways. | 18,931,397 |
Structures of pseudechetoxin and pseudecin, two snake-venom cysteine-rich secretory proteins that target cyclic nucleotide-gated ion channels: implications for movement of the C-terminal cysteine-rich domain. | Cyclic nucleotide-gated (CNG) ion channels play pivotal roles in sensory transduction by retinal photoreceptors and olfactory neurons. The elapid snake toxins pseudechetoxin (PsTx) and pseudecin (Pdc) are the only known protein blockers of CNG channels. These toxins belong to a cysteine-rich secretory protein (CRISP) family containing an N-terminal pathogenesis-related proteins of group 1 (PR-1) domain and a C-terminal cysteine-rich domain (CRD). PsTx and Pdc are highly homologous proteins, but their blocking affinities on CNG channels are different: PsTx blocks both the olfactory and retinal channels with approximately 15-30-fold higher affinity than Pdc. To gain further insights into their structure and function, the crystal structures of PsTx, Pdc and Zn2+-bound Pdc were determined. The structures revealed that most of the amino-acid-residue differences between PsTx and Pdc are located around the concave surface formed between the PR-1 domain and the CRD, suggesting that the concave surface is functionally important for CNG-channel binding and inhibition. A structural comparison in the presence and absence of Zn2+ ion demonstrated that the concave surface can open and close owing to movement of the CRD upon Zn2+ binding. The data suggest that PsTx and Pdc occlude the pore entrance and that the dynamic motion of the concave surface facilitates interaction with the CNG channels. | 18,931,410 |
Crystallization of proteins on functionalized surfaces. | Functionalized mica sheets and polystyrene films exposing ionisable groups have been used as heterogeneous nucleating surfaces for model proteins. Surfaces with different densities of amino or sulfonated groups have been prepared. Crystallization trials were carried out using the hanging-drop vapour-diffusion method. The results show that using these surfaces the starting protein concentration necessary to form crystals is reduced. The effect of these surfaces on the crystallization process may be the consequence of electrostatic interactions between charged residues of proteins and ionisable groups on surfaces. These interactions can be attractive or repulsive, depending on the relative charge of the protein and the surface at the crystallization pH. Both phenomena can induce an increase of the local protein concentration on the surface or in its proximity, favouring nucleation. Moreover, a reduction of the waiting time (an estimation of the nucleation time) was also observed for some proteins, suggesting a surface-stabilization effect on crystal nuclei. | 18,931,412 |
Computing with almost periodic functions. | This paper develops a method for discrete computational Fourier analysis of functions defined on quasicrystals and other almost periodic sets. A key point is to build the analysis around the emerging theory of quasicrystals and diffraction in the setting on local hulls and dynamical systems. Numerically computed approximations arising in this way are built out of the Fourier module of the quasicrystal in question and approximate their target functions uniformly on the entire infinite space. The methods are entirely group theoretical, being based on finite groups and their duals, and they are practical and computable. Examples of functions based on the standard Fibonacci quasicrystal serve to illustrate the method (which is applicable to all quasicrystals modeled on the cut-and-project formalism). | 18,931,421 |
Overexpression, purification, crystallization and preliminary X-ray crystal analysis of Bacillus pallidusD-arabinose isomerase. | D-Arabinose isomerase catalyzes the isomerization of D-arabinose to D-ribulose. Bacillus pallidus D-arabinose isomerase has broad substrate specificity and can catalyze the isomerization of D-arabinose, L-fucose, L-xylose, L-galactose and D-altrose. Recombinant B. pallidus D-arabinose isomerase was overexpressed, purified and crystallized. A crystal of the enzyme was obtained by the sitting-drop method at room temperature and belonged to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 144.9, b = 127.9, c = 109.5 A. Diffraction data were collected to 2.3 A resolution. | 18,931,442 |
Three crystal forms of the bifunctional enzyme proline utilization A (PutA) from Bradyrhizobium japonicum. | Proline utilization A proteins (PutAs) are large (1000-1300 residues) membrane-associated bifunctional flavoenzymes that catalyze the two-step oxidation of proline to glutamate by the sequential action of proline dehydrogenase and Delta(1)-pyrroline-5-carboxylate dehydrogenase domains. Here, the first successful crystallization efforts for a PutA protein are described. Three crystal forms of PutA from Bradyrhizobium japonicum are reported: apparent tetragonal, hexagonal and centered monoclinic. The apparent tetragonal and hexagonal crystals were grown in the presence of PEG 3350 and sodium formate near pH 7. The apparent tetragonal form diffracted to 2.7 A resolution and exhibited pseudo-merohedral twinning such that the true space group is P2(1)2(1)2(1) with four molecules in the asymmetric unit. The hexagonal form diffracted to 2.3 A resolution and belonged to space group P6(2)22 with one molecule in the asymmetric unit. Centered monoclinic crystals were grown in ammonium sulfate, diffracted to 2.3 A resolution and had two molecules in the asymmetric unit. Removing the histidine tag was important in order to obtain the C2 crystal form. | 18,931,443 |
Chromosome polymorphism in Astyanax fasciatus (Teleostei, Characidae). 2--Chromosomal location of a satellite DNA. | Studies about composition of repetitive sequences and their chromosomal location have been helpful to evolutionary studies in many distinct organisms. In order to keep on assessing the possible relationships among different cytotypes of Astyanax fasciatus (Teleostei, Characiformes) in the Mogi-Guaçu River (São Paulo State, Brazil), C-banding, chromomycin A(3) staining, and fluorescent in situ hybridization with a repetitive DNA sequence (As51) isolated from Astyanax scabripinnis were performed in the present work. The constitutive heterochromatin was distributed in terminal regions on long arms of submetacentric, subtelocentric, and acrocentric chromosomes and in the terminal region on short arms of a pair of submetacentric chromosomes in both standard cytotypes. This latter heterochromatic site was also GC-rich, as revealed by chromomycin A(3) staining, corresponding to the nucleolar organizer region (NOR), as shown by previous studies. The sites of the satellite As51 DNA were located in terminal regions on long arms of several chromosomes. Some variant karyotypic forms, which diverge from the two standard cytotypes, also presented distinctive chromosomes carrying As51 satellite DNA. It is possible that the standard 2n = 46 cytotype represents an invader population in the Mogi-Guaçu River able to interbreed with the resident standard 2n = 48 cytotype. Therefore, the variant karyotypes would be related to a possible viable offspring, where complementary chromosomal rearrangements could favor new locations of the satellite DNA analyzed. | 18,931,487 |
Acid production from sugars and sugar alcohols by probiotic lactobacilli and bifidobacteria in vitro. | Some probiotic bacterial strains have been suggested to improve oral health. However, lactobacilli and bifidobacteria are associated with the progression of dental caries. The pH fall caused by 14 probiotic and dairy bacterial strains from glucose, lactose, sucrose, sorbitol and xylitol was followed. All strains used glucose, nine lactose and seven sucrose. Six of the lactobacilli caused a small decrease in pH with sorbitol and two with xylitol. None of the bifidobacteria fermented sugar alcohols. As all the strains could be considered acidogenic, more long-term clinical trials are needed before recommendations for oral health purposes can be made. | 18,931,494 |
Acardiac fetus with large intestine only. A case report. | Acardiac twin syndrome is a rare complication affecting monozygotic twins, where one twin fails to develop normally and completely. In this report, we present an acardiac fetus that was seen for evaluation at 26 weeks of gestation. Initial routine ultrasound examination suggested anomalies. The first detailed ultrasound demonstrated a normal fetus with appropriate growth plus an acardiac twin with a hypoplastic lower limb with subcutaneous edema and intestine-like organ near it. The pregnancy was followed with serial ultrasonography and spontaneous delivery occurred at term. A normal infant was born, and after delivery of the placenta, at the chorionic plate of the placenta there was a sac with diminished fluid, containing some loops of the intestine. A thin cord of one vascular channel was attached to the common placenta. In our literature review, this type of acardiac fetus has not been reported previously. | 18,931,499 |
Assessing genuine parents-offspring trios for genetic association studies. | Family-based association tests such as the transmission disequilibrium test (TDT) are dependent on the successful ascertainment of true nuclear family trios. Relationship misspecification inevitably occurs in a proportion of trios collected for genotyping which undetected can lead to a loss of power and increased Type I error due to biases in over-transmission of common alleles. Here, we introduce a method for evaluating the authenticity of nuclear family trios. Operating in a Bayesian framework, our approach assesses the extent of pedigree inconsistent genotype configurations in the presence of genotyping errors. Unlike other approaches, our method: (i) utilizes information from three individuals collectively (the whole trio) rather than consider two independent pairwise relationships; (ii) down-weighs SNPs with poor performance; (iii) does not require the user to pre-define a rate of genotyping error, which is often unknown to the user and seldom fixed across the different SNPs considered which available methods unrealistically assumed. Simulation studies and comparisons with a real set of data showed that our approach is more likely to correctly identify the presence of true and misspecified trios compared to available software, accurately infers the extent of relationship misspecification in a trio and accurately estimates the genotyping error rates. Assessing relationship misspecification depends on the fidelity of the genotype data used. Available algorithms are not optimised for genotyping technology with varying rates of errors across the markers. Through our comparison studies, our approach is shown to outperform available methods for assessing relationship misspecifications. | 18,931,507 |
Heritability of lung function in severe alpha-1 antitrypsin deficiency. | Severe alpha-1 antitrypsin (AAT) deficiency is a proven genetic risk factor for COPD, but there is marked variation in the development of COPD among AAT deficient subjects. To investigate familial aggregation of lung function in subjects with AAT deficiency, we estimated heritability for forced expiratory volume in 1 s (FEV1) and FEV1/forced vital capacity (FVC) in 378 AAT deficient subjects from 167 families in the AAT Genetic Modifiers Study; all subjects were verified homozygous for the Z AAT deficiency allele. Heritability was evaluated for models that included and excluded an ascertainment correction, as well as for models that excluded, included and were stratified by a cigarette smoking covariate. In models without an ascertainment correction, and in all models without a covariate for smoking, no evidence for familial aggregation of lung function was observed. In models conditioned on the index proband with covariates for smoking, post-bronchodilator FEV1/FVC demonstrated significant heritability (0.26 +/- 0.14, p = 0.03). When we limited the analysis to subjects with a smoking history, post-bronchodilator FEV1 demonstrated significant heritability (0.47 +/- 0.21, p = 0.02). Severity rate phenotypes were also assessed as potential phenotypes for genetic modifier studies. Significant heritability was found with all age-of-onset threshold models that included smoking and ascertainment adjustments. Using the t-distribution, the heritability estimates ranged from 0.43 to 0.64, depending on the age-of-onset of FEV1 decline used for the severity rate calculation. Correction for ascertainment and consideration of gene-by-smoking interactions will be crucial for the identification of genes that may modify susceptibility for COPD in families with AAT deficiency. | 18,931,508 |
[A case of increased resectability with preoperative chemotherapy FOLFOX4 for unresectable rectal cancer]. | The patient was a 57-year-old man. The chief complaints were bleeding upon defecation and decreased body weight. He came to our department in May 2006 because the bleeding had been observed since summer 2005 and he had lost 7 kg in one year. A tumor was palpable on the rectum, approximately 5 cm proximal to the anal verge. Abdominal CT revealed a large tumor within the pelvis and enlarged paraaortic lymph nodes. CEA was 14.0 ng/mL. The patient underwent surgery in June 2006, but the tumor was firmly fixed anterior to the sacrum. We judged it unresectable and performed double-barrel descending colostomy. FOLFOX4 chemotherapy commenced following the surgery, and the tumor marker level normalized following three cycles. Abdominal CT following five cycles showed that the size of the tumor had reduced significantly. Later, the patient developed grade 2 nausea and decreased appetite as adverse events, and the chemotherapy was discontinued at his request. We considered resection possible and performed rectal resection in November 2006. The patient underwent radiotherapy following surgery and is currently receiving S-1. FOLFOX4 may be an effective preoperative chemotherapy for unresectable primary rectal cancer. | 18,931,585 |
[A case of mediastinal cancer of unknown primary with elevation of serum concentrations of interleukin-6]. | Interleukin-6(IL-6)is an inflammatory cytokine. It's produced by monocyte, lymphocyte, vascular endothelial cell, and fibroblast. Elevation of serum concentrations of IL-6 is reported a lot of kind of diseases, such as cancer, infection, inflammatory diseases. But it is production mechanism was still unknown. A 67-year-old man was referred to our hospital because of treatment against the large mediastinal cancer of unknown primary. He had complication with superior vena cave syndrome. After he was exposed to irradiation, the tumor size was reduced, his symptoms were improved and the serum IL-6 level was decreased. | 18,931,593 |
Role of long-chain polyunsaturated fatty acids in the first year of life. | The 2 most abundant long-chain polyunsaturated fatty acids (LCPUFAs) in the brain are docosahexaenoic acid (DHA) and arachidonic acid (ARA), where they have a functional and structural role in infant development. DHA is concentrated in the prefrontal cortex, which is important for association and short-term memory, and in some retinal cells. Concentrations of PUFAs in human breast milk are relatively consistent during the first year of life, and studies have shown that breast-fed infants have a greater mean weight percentage of DHA and a greater proportion of DHA in their red blood cells and brain cortex than formula-fed infants. Furthermore, cortex DHA in breast-fed infants increases with age, probably due to the length of feeding. Maternal supplementation with cod liver oil, which is rich in DHA and eicosapentaenoic acid, improved children's intelligence quotient compared with corn-oil supplementation by 4 years of age. The LCPUFA content of human breast milk is affected by a number of factors, including diet, gestational age, parity, and smoking. Supplementation of formula feed with DHA and ARA results in infant development that is similar to breast-feeding, and may have benefits on blood pressure in later childhood. The beneficial effects of LCPUFA supplementation on visual acuity continue after weaning irrespective of the type of diet. The long-term effects and duration of supplementation of breast- and formula-fed infants requires further investigation. | 18,931,599 |
Consensus recommendations on estrogen receptor testing in breast cancer by immunohistochemistry. | Estrogen receptor (ER) status in breast cancer is currently the most important predictive biomarker that determines breast cancer prognosis after treatment with endocrine therapy. Although immunohistochemistry has been widely viewed as the gold standard methodology for ER testing in breast cancer, lack of standardized procedures, and lack of regulatory adherence to testing guidelines has resulted in high rates of "false-negative" results worldwide. Standardized testing is only possible after all aspects of ER testing--preanalytical, analytical, and postanalytical, have been closely controlled. A meeting of the "ad-hoc committee" of expert pathologists, technologists, and scientists, representing academic centers, reference laboratories, and various agencies, issued standardization testing recommendations, aimed at optimization of clinical ER testing environment, as a step toward improved standardized testing. | 18,931,614 |
Severe metformin intoxication treated with prolonged haemodialyses and plasma exchange. | Biguanides pose a significant risk of morbidity, mortality and permanent sequelae secondary to prolonged periods of hypoglycaemia. To investigate the treatment of massive metformin overdose associated with lactic acidosis. We present the case of a 30-year-old woman, who attempted to commit suicide by ingesting an 85-g massive metformin overdose associated with severe lactic acidosis, which we treated by performing prolonged haemodialysis with bicarbonate and plasma exchange. For the maximum elimination of metformin, extended haemodialysis is required and the treatment of the accompanying metabolic acidosis with bicarbonate is important for the effectiveness of the treatment. Patients benefit much more from the treatment of combined haemodialysis with plasma exchange. | 18,931,616 |
Linking SNPs to CAG repeat length in Huntington's disease patients. | Allele-specific silencing using small interfering RNAs targeting heterozygous single-nucleotide polymorphisms (SNPs) is a promising therapy for human trinucleotide repeat diseases such as Huntington's disease. Linking SNP identities to the two HTT alleles, normal and disease-causing, is a prerequisite for allele-specific RNA interference. Here we describe a method, SNP linkage by circularization (SLiC), to identify linkage between CAG repeat length and nucleotide identity of heterozygous SNPs using Huntington's disease patient peripheral blood samples. | 18,931,668 |
Nedd4 augments the adaptive immune response by promoting ubiquitin-mediated degradation of Cbl-b in activated T cells. | Nedd4 and Itch are E3 ubiquitin ligases that ubiquitinate similar targets in vitro and thus are thought to function similarly. T cells lacking Itch show spontaneous activation and T helper type 2 polarization. To test whether loss of Nedd4 affects T cells in the same way, we generated Nedd4(+/+) and Nedd4(-/-) fetal liver chimeras. Nedd4(-/-) T cells developed normally but proliferated less, produced less interleukin 2 and provided inadequate help to B cells. Nedd4(-/-) T cells contained more of the E3 ubiquitin ligase Cbl-b, and Nedd4 was required for polyubiquitination of Cbl-b induced by CD28 costimulation. Our data demonstrate that Nedd4 promotes the conversion of naive T cells into activated T cells. We propose that Nedd4 and Itch ubiquitinate distinct target proteins in vivo. | 18,931,680 |
Aggressiveness of HNSCC tumors depends on expression levels of cortactin, a gene in the 11q13 amplicon. | 11q13 amplification is a late-stage event in several cancers that is often associated with poor prognosis. Among 11q13-amplified genes, the actin assembly protein cortactin/CTTN is considered a likely candidate for direct involvement in tumor progression because of its cell motility-enhancing functions. We modulated cortactin expression in head and neck squamous cell carcinoma (HNSCC) cell lines. Cortactin expression levels directly correlated with tumor size, vascularization and cell proliferation in an orthotopic HNSCC in vivo model. In contrast, under normal in vitro culture conditions, cortactin expression levels had no effect on cell proliferation. However, cell lines in which cortactin expression was reduced by knockdown (KD) grew poorly in vitro under harsh conditions of growth factor deprivation, anchorage independence and space constraint. In contrast, overexpression of cortactin enhanced in vitro growth under the same harsh conditions. Surprisingly, defects in growth factor-independent proliferation of cortactin-KD cells were rescued by coculture with cortactin-expressing cells. As the cocultured cells are separated by permeable filters, cortactin-expressing cells must secrete growth-supporting autocrine factors to rescue the cortactin-KD cells. Overall, cortactin expression modulates multiple cellular traits that may allow survival in a tumor environment, suggesting that the frequent overexpression of cortactin in tumors is not an epiphenomenon but rather promotes tumor aggressiveness. | 18,931,703 |
[Initial screening of binding-peptide of the cell surface marker CD133 of cancer stem cells]. | To select the binding-peptide of the cell surface marker CD133 of cancer stem cells from phage peptide library, and to find a new tool for research on stem cells, tumor therapy and anti-metastasis of cancer. Biotined mouse CD133 extracellular fraction was used as a target to screen phage 7-peptide library by the high affinity of streptavidin and biotin, and the clones were identified by sandwich ELISA and competitive experiment. Single strand DNA was extracted from these positive clones and was analyzed by single-strand dideoxy-sequencing. After three turn solution panning, five peptides with high affinity shared the same amino acid sequence: APSPMIW and three identical peptides with high affinity shared the same amino acid sequence: LQNAPRS. The peptides that bind with mouse CD133 extracellular fraction with high affinity and specificity were first screened from the phage peptide library for the first time, which initially indicates that the feasibility of screening from phage peptide library with small molecule polypeptide biotined as a target. | 18,931,708 |
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