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Preparation of capsules.
STERILE, SEPARATION
Fresh stool was collected from donors in a sterile plastic container. The dry weight of the stool needed to be more than 100 g. Fresh fecal samples (25%) were mixed with saline (60%) and pharmaceutical-grade glycerol (15%) in a semiautomatic extraction instrument (Treatgut TG-01; Anjiezhishan Company, China). The instr...
PMC10269780
Interventions.
ADVERSE EFFECTS
We obtained informed consent from patients prior to their inclusion in this study. Authorized physicians administered FMT under monitored clinical settings. Before taking capsules orally, patients were required to fast overnight for at least 8 h. All patients received FMT in the morning. Capsules stored at −80°C were r...
PMC10269780
Medication during enrollment.
UC
UC maintenance therapy was provided to patients during FMT. Because patients had a poor response to mesalazine, SASP, or prednisone, stable doses of drugs were used during follow-up after FMT. A subject treated with 5-ASA or SASP continued to receive the same dose of drugs administered during the study. A subject who w...
PMC10269780
Outcomes.
Clinical response at week 12 was defined as a reduction of ≥3 points in the total Mayo score and a decrease of ≥30% from the baseline score (
PMC10269780
The 16S rRNA gene in fecal microbiota and shotgun metagenomic sequencing.
Fresh fecal samples were collected and frozen at −80°C until DNA extraction. According to the manufacturer's protocol, total fecal DNA was extracted from 0.25 g of each homogenized sample using the QIAamp PowerFecal DNA kit (Qiagen). The DNA yield and quality were then checked with a Multiskan GO spectrophotometer (The...
PMC10269780
Microbial bioinformatic analyses.
Raw V3-V4 sequencing reads were assembled to generate high-quality reads using FLASH (
PMC10269780
Serum sample collection and metabolomics.
Serum samples were collected and immediately frozen at −80°C until metabolite extraction. Total metabolite content was extracted from 0.05 g of each of the serum samples, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses, performed using the ultrahigh-performance liquid chromatography (UHPL...
PMC10269780
Statistical analyses and visualization.
Microbial α-diversity indexes, including richness (Observed and Chao1), Shannon diversity (Shannon), and Pielou's evenness (Evenness), were computed using the vegan package (
PMC10269780
Data availability.
The accession numbers of raw sequences have been deposited in the NCBI Sequence Read Archive under BioProject accession no.
PMC10269780
REFERENCES
PMC10269780
Keywords
PMC10570927
Introduction
Young women in sub-Saharan Africa are at a high risk of unintended pregnancies [
PMC10570927
Methods
PMC10570927
Study design
Within the ECHO trial (
PMC10570927
STI and BV testing
Women were tested for
PMC10570927
RNA-Seq library preparation and quality control analysis of data
Cervical cytobrush samples collected at baseline and at one-month follow-up visit from 188 matched participants (
PMC10570927
Differential expression analysis
A paired differential expression analysis, i.e. post-vs-pre (one-month vs. baseline) comparison, was carried out for each study arm. For the paired analyses, only protein-coding probe-target genes were retained: hg19 probe coordinates from the Illumina RNA Exome kit were cross-mapped to hg38 genome assembly using cross...
PMC10570927
Defining Lactobacillus-dominant versus Lactobacillus-depleted vaginal microbiota at baseline in DMPA-IM study arm
To investigate if the cervicovaginal transcriptomic changes observed in women using DMPA-IM were associated with a
PMC10570927
Deconvolution of immune cell types using RNA-seq data of women assigned to cu-IUD
The normalized gene counts of samples in Cu-IUD study arm (
PMC10570927
Cell culture and luminex
REGRESSION
Human Endocervical Epithelial cells (END1/E6E7) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured in keratinocyte serum-free medium (KSFM; Gibco Life Technologies, Grand Island, NY, USA) with 0.1 ng/ml human epidermal growth factor, 0.05 mg/ml bovine pituitary extract, 0.4 ...
PMC10570927
Statistical analysis
All downstream statistical analyses were performed in RStudio. Differences in study population characteristics according to study arm were tested using Pearson's Chi-squared test or Fisher's exact test (when the expected value was <5) for count data and unpaired Student's
PMC10570927
Results
PMC10570927
Cohort characteristics
For this nested mucosal sub-study of the ECHO trial, we included a total of 188 women with matched endocervical cytobrush samples obtained prior to, and one month after contraceptive initiation (DMPA-IM, Demographics and metadata of ECHO subjects recruited to substudy of mucosal transcriptomics.BMI; body mass index, KE...
PMC10570927
Cu-IUD, LNG implant and DMPA-IM initiation induce distinct patterns of endocervical gene-expression
To assess the impact of initiation of contraception on the endocervical transcriptome, we identified DEGs induced after one month of contraception usage relative to baseline within each study arm in a longitudinal intention-to-treat (ITT) analysis (Supplementary Table 2). A total of 151 DEGs were induced by DMPA-IM (60...
PMC10570927
DMPA-IM initiation induced an enrichment of T cell gene signatures in the FGT
RECRUITMENT
The use of DMPA-IM has been reported to induce a diverse array of biological effects in the FGT. A closer examination of the 151 DEGs identified in DMPA-IM arm (Supplementary Table 2) showed significant alterations in a handful of genes that function in T cell activation (PRF1, TNFSF18, MAF) and tissue recruitment of i...
PMC10570927
DMPA-IM induced transcriptomic alterations are not strongly driven by microbial composition
We performed a sub-analysis to investigate whether the transcriptomic changes observed within the DMPA-IM arm were specific to the vaginal microbiota of participating women, i.e.
PMC10570927
Growth factor signaling and epithelial barrier pathways enriched in women assigned to LNG implant
PROLIFERATION, ADHESION
Within the LNG implant arm, several pathways associated with cell proliferation and growth including growth factor signaling by epidermal growth factor (EGF) and transforming growth factor (TGF)-β, Notch signaling, and WNT signaling, as well as cell-cell adhesion pathways were positively enriched, whereas pathways asso...
PMC10570927
Cu-IUD usage causes significant perturbations to the cervicovaginal transcriptome
Cu-IUD treatment, with 907 DEGs identified, had a dramatic impact on gene expression (Immune activation pathways enriched in women assigned to Cu-IUD (nominal p-value < 0.1). Important pathways enriched in Cu-IUD study arm, were selected. (A) A dot plot to represent the gene set enrichment analysis (GSEA) statistics fo...
PMC10570927
Cu-IUD initiation recruits innate immune cells into the endocervix
Cu-IUD initiation
Since Cu-IUD initiation resulted in the highest significant changes in the FGT gene expression as compared to LNG implant and DMPA-IM, its influence on the immune cell populations was further investigated using digital flow cytometry methods (Cu-IUD initiation induces significant changes in the elements of innate immun...
PMC10570927
Copper ions do not exert inflammatory effects on endocervical epithelial cells
EVENTS
To test if the inflammatory events observed after Cu-IUD initiation in vivo were due to a direct effect of CuThe concentrations of multiple endocervical cytokines are altered by biologically relevant copper (Cu
PMC10570927
Discussion
inflammation
STIS, INFLAMMATION, PROLIFERATION, INFILTRATING
Although, sub-Saharan African women are at high risk for both STIs, including HIV, and unintended pregnancies, the impact of commonly used NBC methods on the mucosal immune environment has not been studied in detail using randomized designs. In this study, we examined the effects of three common and highly effective NB...
PMC10570927
Conclusions
These results illustrate the variable influence of different contraceptives on the FGT host environment with Cu-IUD insertion resulting in the highest perturbation of the endocervical transcriptome, mainly related to immune responses, as compared to DMPA-IM and LNG implant initiation. DMPA-IM use led to an enrichment o...
PMC10570927
Table S1
DESeq results for Chlamydia-based analysis
PMC10570927
Table S2
DESeq results for paired-analysis (post-vs-pre) for each study-arm in a longitudinal intention-to-treat (ITT) analysis.
PMC10570927
Table S3
Custom in-house gene-sets used for GSEA.
PMC10570927
Table S4
DESeq results for Lacto-dominance(LD)/non lacto-dominance (nonLD) at baseline, paired-analysis
PMC10570927
Table S5
DESeq results for Lacto-dominance(LD)/non lacto-dominance (nonLD) at baseline, paired-analysis
PMC10570927
Table S6
MOS
Differentially expressed pathways in the DMPA-IM group, pre-treatment vs 1 mos post-treatment, separated by women with Lactobacillus-dominated vaginal microbiome (LD), Lactobacillus-depleted vaginal microbiota (non-LD) or both groups analyzed together (LD + nonLD).
PMC10570927
Table S7
Influence of Cu-IUD initiation on the population of vaginal immune cells
PMC10570927
Supplementary material
TD
Supplementary Figure 1: A schematic representation of the RNA-Seq workflow. (A) The flowchart of RNA-Seq analysis outlines the experimental and data analysis steps carried out in the study. (B) The number of samples at collection timepoints and after quality-based filtering in each study arm.Supplementary Figure 2: Qua...
PMC10570927
Funding
This work and the Evidence for Contraceptive Options and HIV Outcomes (ECHO) Study were made possible by the combined generous support of the Bill & Melinda Gates Foundation (Grant OPP1032115), the American people through the United States Agency for International Development (Grant AID-OAA-A-15–00045), the Swedish Int...
PMC10570927
References
PMC10570927
Data availability
Transcriptomic data is available in the Gene Expression Omnibus (GEO) repository under accession number GSE190923. Custom R scripts and supporting documentation on the RNA-Seq analyses are available at
PMC10570927
Background
Sexting refers to the exchange of sexually explicit digital content in the form of texts, photos, or videos. In recent years, sexting has become a public health concern. Surveys in Malaysia show a high prevalence of young adults engaged in sexting. Given that sexting is associated with sexual risk behavior, cyberbullyi...
PMC10625075
Objective
SECONDARY
This study aimed to develop and implement a sexting intervention module guided by the prototype willingness model (PWM), delivered using web-based animated video, and evaluate its effectiveness among diploma students from a public higher educational institution. The primary outcomes were intention and willingness to se...
PMC10625075
Methods
This 2-armed, parallel, single-blinded cluster randomized controlled trial was conducted in a public higher educational institution in the state of Melaka, Malaysia. Diploma students from 12 programs were randomly allocated into intervention and control groups. Both groups answered a self-administered web-based questio...
PMC10625075
Results
There were a total of 300 participants with an attrition rate of 8.3% (n=25). After adjusting for age, sex, relationship status, and the amount of time spent on the web, there were significant differences in the intention to sext (β=–.12;
PMC10625075
Conclusions
In this study, the sexting intervention module using the PWM that was delivered via web-based animated videos was effective in reducing intention and willingness to sext as well as in improving knowledge of sexting, attitudes, perceived norms, and prototype perceptions. Therefore, relevant agencies involved in the prom...
PMC10625075
Trial Registration
Thai Clinical Trial Registry TCTR20201010002; https://www.thaiclinicaltrials.org/show/TCTR20201002001
PMC10625075
Introduction
PMC10625075
Background
Sexting refers to the exchange of sexually explicit digital content in the form of texts, photos, or videos [In recent years, sexting has emerged as a major public health concern in Malaysia. According to the literature, the prevalence of sexting among Malaysian youths was higher than in Western nations [Moreover, sext...
PMC10625075
Prior Work
To date, several observational studies have asserted that the behavior of sexting is preceded by an intention and willingness to sext [In this digital age, animated videos are widely acknowledged as an effective teaching tool, especially for students in higher educational institutions (HEIs) [
PMC10625075
Objective
This study aimed to develop, implement, and evaluate the effects of the newly developed sexting intervention module (SIM) delivered via web-based animated videos on the intention and willingness to sext among diploma students. We hypothesized that the intervention would reduce the intention and willingness to sext amon...
PMC10625075
Methods
PMC10625075
Study Design
This study was a 2-armed, parallel, single-blinded, cluster randomized controlled field trial. The available program on the main campus was chosen as a cluster unit with the assumption that students from a specific program only interact with those within the same program as they only engaged in web-based learning at ho...
PMC10625075
Setting and Recruitment
HIV infections
HIV INFECTIONS
This study was conducted in one of the public HEIs in Melaka state, Malaysia. Melaka state was selected as the study site as it is one of the cities in the Association of Southeast Asian Nations that has committed to achieving 0 new HIV infections by 2030 [The study was conducted on the main campus of the largest HEI i...
PMC10625075
Randomization and Allocation Concealment
The 12 programs on the main campus were randomly allocated into intervention and control groups at a ratio of 1:1. All programs were number-coded, and subsequently, a simple randomization process was conducted using Random.Org software (Random.Org LLC) [
PMC10625075
Sample Size Calculation
The sample size was calculated to detect a mean difference of 0.44 (SD 2.19) in an intention toward sexting between the intervention and control groups based on the closest variable to this study [
PMC10625075
Intervention Module
The newly developed intervention module, SIM, used 3 constructs of PWM, including attitude, perceived norms, and prototype perceptions. It was developed in Malay, which is the local native language. The module consisted of information regarding the definition of sexting, the negative impact of sexting, the prevalence o...
PMC10625075
Outcomes
PMC10625075
Overview
SECONDARY
A validated self-administered questionnaire was used to measure the sociodemographic characteristics and the outcomes of this study. The questionnaire was divided into 2 sections. The first section captured the respondent’s age, sex, relationship status, and the amount of time spent on the web. The questions in the sec...
PMC10625075
Primary Outcome Measures
The intention to sext was measured using a 15-item questionnaire with a 6-point Likert scale adapted from a previous study [
PMC10625075
Secondary Outcome Measures
An 8-item questionnaire was used to measure the knowledge of sexting with multiple-choice responses: “yes,” no,” or “I don’t know” [
PMC10625075
Data Collection
MAY
Data were collected at 3 different time points, namely, baseline, immediate postintervention, and 3 months postintervention starting from December 2021 to May 2022. Due to the COVID-19 pandemic, data collection was performed via a web-based Google form. The link was sent to all the participants, and they were given 1 w...
PMC10625075
Data Analysis
Data analysis was performed with SPSS (version 26.0; IBM Corp). The participants were analyzed based on their initial assigned group. Data cleaning and error checking were performed before analysis. Median and IQR were used to summarize the nonnormally distributed data for continuous variables, while frequencies and pe...
PMC10625075
Ethics Approval
This study was approved by the Ethics Committee for Research Involving Human Subjects Universiti Putra Malaysia (JKEUPM-2020-321). The HEI administration and head of program approval as well as the respondents’ written consent were obtained before the study.
PMC10625075
Results
PMC10625075
Discussion
PMC10625075
Principal Findings
SECONDARY
To the best of our knowledge, this is the first RCT to evaluate the effectiveness of a theory-based SIM delivered via web-based animated videos on the intention and willingness to sext among diploma students. The secondary outcomes assessed in this study included knowledge, attitude, perceived norms, and prototype perc...
PMC10625075
Strengths and Limitations
multiple-risk behaviors
To date, interventional research on sexting remains scarce. Therefore, this study represents a critical starting point for public health interventions aimed at addressing sexting-related issues in Malaysia. The use of theory-based intervention and web-based animated videos can be seen as a new direction for future publ...
PMC10625075
Conclusions
SEXUALLY TRANSMITTED INFECTIONS
The PWM-based intervention module delivered via web-based animated videos in this study was effective in reducing the intention and willingness to sext apart from improving the knowledge of sexting, attitude, perceived norms, and prototype perceptions among diploma students. This intervention can be implemented by rele...
PMC10625075
Abbreviations
Consolidated Standards of Reporting Trialsgeneralized linear mixed modelhigher educational institutionprototype willingness modelrandomized controlled trialsexing intervention modulesexual and reproductive healththeory of planned behaviortheory of reasoned action
PMC10625075
Data Availability
The data sets generated during and/or analyzed during this study are available from the corresponding author on reasonable request.
PMC10625075
Graphical abstract
PMC10189551
Keywords
PMC10189551
Abbreviations
SEPARATION
area under the ROC curvelogistic regressionMinamata diseasemagnetoencephalographymethylmercurymagnetic resonance imagingoversampled temporal projectionreceiver operating characteristic curvesomatosensory evoked magnetic fieldssomatosensory evoked potentialsprimary somatosensory cortexsecondary somatosensory cortexsigna...
PMC10189551
Introduction
neurological damage, neurologic signs, damages, ataxia
CONSTRICTION, SENSORY DISTURBANCE, AUDITORY DISTURBANCES, SENSORY DISTURBANCES
Methylmercury (MeHg) is a major environmental neurotoxicant that damages the central nervous system depending on the dose and duration of exposure (MD is classified based on the onset time as acute/subacute MD (manifestation during the heavy contamination of seafood with MeHg in the Minamata Bay) and chronic MD (manife...
PMC10189551
Materials and methods
PMC10189551
Participants
±
RECRUITMENT, MINAMATA DISEASE
Between April 2010 and October 2022, we recruited 42 patients with MD (20 women, 22 men; mean age 70.0 ± 10.6 years) and 289 healthy volunteers (131 women, 158 men; mean age 67.7 ± 9.4 years). Flowchart of participant recruitment and enrollment. MD = Minamata disease.
PMC10189551
MEG recording
The right and left median nerves were stimulated at the wrist in separate recordings using constant current pulses of 0.2-ms duration. Stimulus intensity determined by visual inspection was 1.5 times the motor threshold to evoke twitches of the thumb. The interstimulus interval was 3.7 s to avoid habituation to SII res...
PMC10189551
MR image acquisition
The three-dimensional (3D) T1-weighted images were acquired using a 1.5 T MRI unit (Magnetom Symphony, Siemens Healthcare, Erlangen, Germany) or a 3 T MRI unit (Discovery MR750 3.0 T, GE Healthcare, Milwaukee, WI).
PMC10189551
Data acquisition
peripheral sensory nerve impairment
SENSORY DISTURBANCE
We recorded the SEF responses using a whole-head 306-channel sensor array (Vectorview, ELEKTA Neuromag, Helsinki) that comprises 204 planar gradiometers and 102 magnetometers. We analyzed the MEG data recorded by 204-channel planar-type gradiometers. Before the recording, three anatomical fiducial points (nasion and bi...
PMC10189551
Data processing
Because this study focused on the response of N20m, we applied a band pass filter (6–200 Hz) to the preprocessed data using MNE-Python (v0.20) (We compared neural responses (P20m rate and non-response rate) and early somatosensory cortical processing (N20m amplitude, cross-correlation value, and gamma response) between...
PMC10189551
Statistical analyses
Python machine learning library
For each MEG parameter, the non-parametric Mann–Whitney The receiver operating characteristic (ROC) curve describes a diagnostic trade-off between sensitivity and specificity with the adjustment of a discriminative threshold (All tests were two-tailed, and statistical significance was set at a p-value of 0.05. All anal...
PMC10189551
Ethical declarations
Minamata Disease, NIMD
MINAMATA DISEASE
This study was conducted in accordance with the ethical principles of the Declaration of Helsinki. This study was approved by the Ethics Committee of the National Institute for Minamata Disease (NIMD 10/001, 13/001, 15/001, 16/003, 22/002). Participation was voluntary. Written informed consent was obtained from all par...
PMC10189551
Results
PMC10189551
Evaluation of peripheral sensory nerve function
±
The conduction velocity of the median nerve SNAPs was significantly slower in patients with MD than in controls on either side (left side: control: 53.7 ± 4.9 m/s; MD: 50.9 ± 5.0 m/s; p = 0.0001; right side: control: 53.6 ± 5.1 m/s; MD: 50.8 ± 5.1 m/s; p = 0.0002), despite adjustments for multiple comparisons via Bonfe...
PMC10189551
Neural responses and N20m amplitude
MINAMATA DISEASE
The P20m rate and non-response rate were significantly higher in patients with MD than in controls on either side, despite adjustments for multiple comparisons via Bonferroni correction (p < 0.0001) (Comparisons of MEG findings between controls and patients with MD. Patients with MD demonstrate significantly higher non...
PMC10189551
Single-trial analysis of P20m and absent N20m peak
We examined the frequency of P20m epochs, N20m amplitude of averaged SEFs in N20m epochs, and P20m amplitude of averaged SEFs in P20m epochs in controls and patients with MD with the P20m or an absent N20m peak. The controls displayed fewer P20m epochs, and the averaged sensor SEFs and averaged SEFs of the N20m epochs ...
PMC10189551
Cross-correlation value of N20m
We observed a significant reduction of the cross-correlation value in patients with MD on either side, despite adjustments for multiple comparisons via Bonferroni correction (p < 0.0001) (Time-frequency analysis of SEFs. Compared with that in controls, the power strength of induced gamma oscillations was significantly ...
PMC10189551
Time-frequency analysis of SEFs
Compared with that in controls, the power strength of induced gamma oscillations decreased on either side in patients with MD (
PMC10189551
Correlations between two SEF parameters
The cross-correlation value and power strength of induced gamma oscillations on either side exhibited the highest negative correlation with the non-response rate and P20m rate (Correlations between two SEF parameters. On either side, the cross-correlation value exhibited the highest correlation with the non-response ra...
PMC10189551
Discrimination analysis
REGRESSION
The AUC was > 0.77 (range; 0.77–0.79) for all parameters and was the highest (0.80 [range; 0.72–0.88]) for the integrated SEF parameters using the LogReg method (Receiver operating characteristic curves and corresponding area under the curve values for SEF parameters. The AUC was > 0.77 for all parameters, and the AUC ...
PMC10189551
Discussion
chronic-phase central somatosensory disturbance, neuronal synchronization, bleeding, stroke, neoplasms, Minamata disease, infarction
DISORDER, BRAIN ATROPHY, BLEEDING, STROKE, DYSPLASTIC, NEOPLASMS, DISEASES, PATHOPHYSIOLOGY, DISORDERS, BRAIN TUMORS, MULTIPLE SCLEROSIS, UNILATERAL POLYMICROGYRIA, DEMYELINATION, MINAMATA DISEASE, INFARCTION
We performed single-trial analysis of SEFs to explore the pathophysiological mechanisms of central somatosensory disturbance in patients with chronic MD. Detailed analyses of SEFs under restricted vicinity of the N20m (peak latency ± 3.0 ms) characterized the altered SI response and early somatosensory cortical process...
PMC10189551
Conclusions
Functional abnormalities in early somatosensory cortical processing (reduced N20m amplitude, cross-correlation value, and SI gamma responses) as well as impaired SI responses (increased non-response rate and P20m rate) can be clearly detected in patients with chronic MD using MEG. Single-trial neuromagnetic analysis of...
PMC10189551
Funding
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
PMC10189551
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
PMC10189551
References
PMC10189551
Supplementary data
The following are the Supplementary data to this article:
PMC10189551
Supplementary data 1
PMC10189551