Inconsistent Protocols Research Articles

KNOWLEDGE, ATTITUDE AND PRACTICES OF INTENSIVE CARE UNIT NURSES ABOUT PHYSICAL RESTRAINT AT TERTIARY CARE HOSPITAL LAHORE

Physical restraint is a common intervention in intensive care units (ICUs) to ensure patient safety and prevent harm. However, its inappropriate use poses ethical and clinical challenges, particularly in resource-constrained healthcare systems like Pakistan. Objective: To assess the knowledge, attitudes, and practices of ICU nurses regarding physical restraint in a tertiary care hospital in Lahore, Pakistan. Methods: A descriptive cross-sectional study was conducted among 137 ICU nurses using a structured questionnaire. The data were analyzed using SPSS version 26, with results presented as frequencies and percentages. Results: The study revealed that 47.1% of nurses demonstrated moderate knowledge of physical restraint, while 33.1% exhibited good knowledge. Attitudes were mixed, with 52.2% displaying positive attitudes and 47.8% exhibiting negative attitudes. Good practices were reported by 61.8% of nurses, whereas 38.2% showed poor adherence to recommended protocols. Nurses with higher qualifications and greater experience demonstrated significantly better knowledge and practices. Conclusion: Despite moderate knowledge and adherence to some practices, gaps in attitudes and inconsistent protocol adherence remain challenges in ICU settings. Targeted education, institutional support, and clear policies are essential to enhance nurses' competency in the ethical and effective use of physical restraints.

Anti-diabetic therapies on dental implant success in diabetes mellitus: a comprehensive review.

Dental implant therapy faces challenges in patients with Type 1 and Type 2 Diabetes Mellitus (T1DM and T2DM) due to adverse effects on bone metabolism and immune response. Despite advancements, diabetic patients face higher risks of peri-implantitis and compromised osseointegration. This review assesses the impact of anti-diabetic medications on implant outcomes, offering insights to bridge the gap between animal studies and clinical practice. By evaluating pharmacotherapeutic strategies in preclinical models, this review guides future research designs to improve implant success rates in diabetic individuals. A comprehensive literature review identified 21 animal studies examining the impact of anti-diabetic medications on dental and bone implants. These studies explored diabetes models, medication regimens, and designs to assess outcomes related to bone metabolism, osseointegration, and peri-implant tissue responses. The findings are systematically summarized, highlighting the scope, design, and procedures of each study. An example includes placing a dental implant in the molar region of a mouse, providing insight into preclinical approaches. Twenty-one animal studies, primarily using rodents, investigate various anti-diabetic medications on dental and bone implants. Interventions include insulin, aminoguanidine, voglibose, sitagliptin, exenatide, and metformin, analyzing outcomes like bone-implant contact (BIC), bone volume (BV), and counter-torque values in T1DM and T2DM models. The impacts of these medications on implant osseointegration under diabetic conditions are detailed, with their benefits and shortcomings assessed. The findings and challenges of existing animal studies on diabetes mellitus (DM) and implant osseointegration are presented. Despite T2DM prevalence, research primarily focuses on T1DM models due to easier experimental practicalities, limiting applicability. Inconsistent protocols in studies compromise reliability regarding anti-diabetic treatments' effectiveness on osseointegration. Standardized methodologies and long-term assessments of local drug delivery alongside systemic anti-DM treatments are crucial to manage DM-related complications in implant dentistry. Insulin administration in short-term T1DM animal studies enhances implant osseointegration. However, the efficacy of non-insulin medications remains inconclusive. Rigorous experimental designs are needed to address inconsistencies and assess long-term impacts. Larger-sized (e.g., porcine) animal studies across various intraoral implant scenarios are required. Future research should focus on enhancing clinical applicability and improving implant stability in evolving conditions.

Evidence Synthesis and Clinical Recommendations for Supporting School Students With Sensory Processing Challenges: A Rapid Review.

Children with sensory processing challenges often need supports to access, participate in, and achieve at school. However, research on best practice is varied, presenting difficulty for practitioners to assess the appropriateness of each support. To provide evidence-informed recommendations regarding best practices in school-based supports for students with sensory processing challenges via a Rapid Evidence Assessment of current literature. Searches were run in CINAHL Complete, Education Research Complete, ERIC, MEDLINE, PsycINFO, and Scopus (for articles published 2010 to 2023). Informed by a 2010 review, articles citing publications identified in the prior review were also included. A reviewer screened the title and abstract of each article for inclusion, and another screened the full texts, with 10% of each reviewer's articles assessed by the other. Interrater reliability was excellent. The review included 61 articles, with only 15% constituting good evidence (38% provided adequate evidence, 31% provided poor evidence, and 16% were reviews). No support could be wholly recommended without caveats, but there was some evidence that sound amplification systems, alternate seating, and multisensory environments may be beneficial in certain conditions or with certain populations. The use of weighted vests is not recommended. Most research had small sample sizes and inconsistent protocols, highlighting the importance of ongoing research. Evidence-informed practitioners should review research findings carefully and integrate evidence from comprehensive clinical assessment and understanding of the child, school, and family contexts when assessing the appropriateness of a support for each student. Plain-Language Summary: Children with sensory processing challenges often need supports to access, participate in, and achieve at school. Occupational therapists provide school-based supports for children with sensory processing challenges with the goal of enabling access, participation, and achievement. It is difficult, however, for practitioners to assess the appropriateness of each support because research on best practice varies. This review of the current literature provides practitioners with evidence on supports for school children with sensory processing challenges to inform decision-making and the selection of appropriate supports for their clients. When assessing the appropriateness of a support for a student, practitioners should carefully review current research findings and integrate evidence from comprehensive clinical assessments along with an understanding of the student, school, and family contexts.

The Effectiveness of Auricular Acupressure on Chronic Musculoskeletal Pain: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

Objectives: To assess the effectiveness of auricular acupressure (AA) in managing pain and disability in individuals with chronic musculoskeletal pain. Materials and Methods: A systematic search on six electronic databases was performed from their inception to May 7, 2023, to identified relevant randomized controlled trials (RCTs). Two independent reviewers screened the abstracts and full texts, extracted data, and assessed risk of bias using RoB 2. The primary outcomes were pain intensity and disability. The secondary outcomes were pain pressure thresholds, pain catastrophizing level, and fear avoidance beliefs. A random-effects model was used for meta-analyses. The certainty of evidence was evaluated using the Grading of Recommendations Assessment, Development, and Evaluation. Sensitivity analyses were conducted after removing low-quality papers. Results: Of 633 identified records, six studies involving 496 participants were included. All included studies compared the effectiveness of AA with sham controls in treating various chronic musculoskeletal pain. Four meta-analyses were conducted to compare the effectiveness of AA with sham controls. Low-quality evidence supported that AA had a large effect size on postintervention subjective pain reduction (standardized mean difference [SMD] = -0.95; 95% confidence interval [CI]: -1.36 to -0.54; p = 0.00; I2 = 52.61%); moderate-quality evidence substantiated that AA had a large effect size on enhancing postintervention pressure pain threshold (SMD = -0.55; 95% CI: -0.88 to -0.23; p = 0.00; I2 = 0%). There was low-quality evidence that AA had a large effect on reducing postintervention disability (SMD = -0.68; 95% CI: -1.24 to -0.12; p = 0.02; I2 = 51.33%). Our sensitivity analysis reaffirmed the same conclusion regarding pain reduction immediately after the intervention. Fourteen participants reported minimal adverse events, including soreness, tenderness, irritation, and redness, which disappeared within 1-7 days. Discussion: Our systematic review revealed that AA significantly improved pain, pressure pain thresholds, and disability in individuals with various chronic musculoskeletal pain conditions immediately post-treatment compared with sham treatment. Given the paucity of studies and inconsistent protocols, future RCTs are warranted to evaluate the effectiveness of AA in people with chronic musculoskeletal pain at a longer follow-up with detailed protocols, which allows researchers and clinicians to optimize AA intervention. Conclusion: AA has immediate post-treatment benefits for chronic musculoskeletal pain, whereas its effects at the 1- or 6-month follow-up remain uncertain.

Serum Glycome as a Diagnostic and Prognostic Factor in Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM) is a risk factor for both mother and fetus/neonate during and after the pregnancy. Inconsistent protocols and cumbersome screening procedures warrant the search for new and easily accessible biomarkers. We investigated a potential of serum N-glycome to differentiate between healthy pregnant women (n = 49) and women with GDM (n = 53) using a lectin-based microarray and studied the correlation between the obtained data and parameters of glucose and lipid metabolism. Four out of 15 lectins used were able to detect the differences between the control and GDM groups in fucosylation, terminal galactose/N-acetylglucosamine (Gal/GlcNAc), presence of Galα1,4Galβ1,4Glc (Gb3 antigen), and terminal α2,3-sialylation with AUC values above 60%. An increase in the Gb3 antigen and α2,3-sialylation correlated positively with GDM, whereas the amount of fucosylated glycans correlated negatively with the content of terminal Gal/GlcNAc. The content of GlcNAc oligomers correlated with the highest number of blood analytes, indices, and demographic characteristics, but failed to discriminate between the groups. The presence of terminal Gal residues correlated positively with the glucose levels and negatively with the LDL levels in the non-GDM group only. The results suggest fucosylation, terminal galactosylation, and the presence of Gb3 antigen as prediction markers of GDM.

Challenges of Driver Drowsiness Prediction: The Remaining Steps to Implementation

Driver drowsiness has caused a large number of serious injuries and deaths on public roads and incurred billions of taxpayer dollars in costs. Hence, monitoring of drowsiness is critical to reduce this burden on society. This paper surveys the broad range of solutions proposed to address the challenges of driver drowsiness, and identifies the key steps required for successful implementation. Although some commercial products already exist, with vehicle-based methods most commonly implemented by automotive manufacturers, these systems may not have the level of accuracy required to properly predict and monitor drowsiness. State-of-the-art models use physiological, behavioural and vehicle-based methods to detect drowsiness, with hybrid methods emerging as a superior approach. Current setbacks to implementing these methods include late detection, intrusiveness and subject diversity. In particular, physiological monitoring methods such as Electroencephalography (EEG) are intrusive to drivers; while behavioural monitoring is least robust, affected by external factors such as lighting, as well as being subject to privacy concerns. Drowsiness detection models are often developed and validated based on subjective measures, with the Karolinska Sleepiness Scale being the most popular. Subjective and incoherent labelling of drowsiness, lack of on-road data and inconsistent protocols for data collection are among other challenges to be addressed to progress drowsiness detection for reliable on-road use.

Towards a standardized multi-tissue decellularization protocol for the derivation of extracellular matrix materials.

The goal of tissue decellularization is to efficiently remove unwanted cellular components, such as DNA and cellular debris, while retaining the complex structural and molecular milieu within the extracellular matrix (ECM). Decellularization protocols to date are centered on customized tissue-specific and lab-specific protocols that involve consecutive manual steps which results in variable and protocol-specific ECM material. The differences that result from the inconsistent protocols between decellularized ECMs affect consistency across batches, limit comparisons between results obtained from different laboratories, and could limit the transferability of the material for consistent laboratory or clinical use. The present study is the first proof-of-concept towards the development of a standardized protocol that can be used to derive multiple ECM biomaterials (powders and hydrogels) via a previously established automated system. The automated decellularization method developed by our group was used due to its short decellularization time (4 hours) and its ability to reduce batch-to-batch variability. The ECM obtained using this first iteration of a unified protocol was able to produce ECM hydrogels from skin, lung, muscle, tendons, cartilage, and laryngeal tissues. All hydrogels formed in this study were cytocompatible and showed gelation and rheological properties consistent with previous ECM hydrogels. The ECMs also showed unique proteomic composition. The present study represents the first step towards developing standardized protocols that can be used on multiple tissues in a fast, scalable, and reproducible manner.

Nurses' perceptions and use of near infrared spectroscopy in paediatric cardiac intensive care.

Near-infrared spectroscopy (NIRS) is a non-invasive technology that estimates regional oxygen saturation. Literature demonstrates that NIRS can provide valuable data for clinical staff. However, little research has addressed the nursing care and management of NIRS in the critical care environment. To assess nurses' perception around the use of NIRS and current NIRS practice within paediatric cardiac intensive care unit (PCICUs). A 53-item cross-sectional electronic survey was developed to assess indications for NIRS, critical value thresholds and interventions, barriers to use, policies and procedures, and nursing perceptions. Descriptive statistics were used to summarize and aggregate data. Among the 28 responding sites (63.6% response rate), usage of NIRS was variable and patient-dependent. Most nurses reported using NIRS in patients with unstable physiology such as post-operative single ventricle (n=25, 89.3%) and concern for shock (n=21, 75.0%). Critically low cerebral values varied among respondents from less than 40 (n=3, 10.7%) to less than 60 (n=4, 14.3%), with lower critical values permitted for single ventricle physiology: less than 40 (n=8, 28.6%) to less than 50 (n=6, 21.4%). Reported barriers to using NIRS included skin breakdown (n=9, 32.1%), lack of consistency in decision-making among physicians (n=13, 46.4%), and not using NIRS data when developing a plan of care (n=11, 39.3%). Most (n=24, 85.7%) nurses reported that NIRS provided valuable information and was perceived to be beneficial for patients. NIRS monitoring is a common technology in the care of complex congenital heart disease patients. Most nurses valued this technology, but inconsistencies and practicalities around its use in guiding patient management were found to be problematic. NIRS is commonly used in the PCICU and although nurses perceived NIRS to be useful for their practice, the variability in the interpretation of values and inconsistent protocols and decision-making by physicians was challenging.

Computed Tomography Angiography (CTA) in Selected Scenarios with Risk of Possible False-Positive or False-Negative Conclusions in Diagnosing Brain Death.

It is widely accepted that brain death (BD) is a diagnosis based on clinical examination. However, false-positive and false-negative evaluation results may be serious limitations. Ancillary tests are used when there is uncertainty about the reliability of the neurologic examination. Computed tomography angiography (CTA) is an ancillary test that tends to have the lowest false-positive rates. However, there are various influencing factors that can have an unfavorable effect on the validity of the examination method. There are inconsistent protocols regarding the evaluation criteria such as scoring systems. Among the most widely used different scoring systems the 4-point CTA-scoring system has been accepted as the most reliable method. Appropriate timing and/or Doppler pre-testing could reduce the number of possible premature examinations and increase the sensitivity of CTA in diagnosing cerebral circulatory arrest (CCA). In some cases of inconclusive CTA, the whole brain computed tomography perfusion (CTP) could be a crucial adjunct. Due to the increasing significance of CTA/CTP in determining BD, the methodology (including benefits and limitations) should also be conveyed via innovative electronic training tools, such as the BRAINDEXweb teaching tool based on an expert system.

Factors influencing the mental health of an ethnically diverse healthcare workforce during COVID-19: a qualitative study in the United Kingdom

ABSTRACT Background: Healthcare workers (HCWs) have been reported to be experiencing a deterioration in their mental health due to COVID-19. In addition, ethnic minority populations in the United Kingdom are disproportionately affected by COVID-19. It is imperative that HCWs are appropriately supported and protected from mental harm during the pandemic. Our research aims to add to the evidence base by providing greater insight into the lived experience of HCWs from diverse ethnic backgrounds during the pandemic that had an impact on their mental health. Methods: We undertook a qualitative work package as part of the United Kingdom Research study into Ethnicity And COVID-19 outcomes among Healthcare workers (UK-REACH). As part of the qualitative research, we carried out 16 focus groups with a total of 61 HCWs between December 2020 and July 2021. The aim of the study was to explore topics such as their experiences, fears and concerns, while working during the pandemic. The purposive sample included ancillary healthcare workers, doctors, nurses, midwives and allied health professionals from diverse ethnic backgrounds to ensure inclusion of underrepresented and disproportionately impacted individuals. We conducted discussions using Microsoft Teams. Recordings were transcribed and thematically analysed. Results: Several factors were identified which impacted on the mental health of HCWs during this period including anxiety (due to inconsistent protocols and policy); fear (of infection); trauma (due to increased exposure to severe illness and death); guilt (of potentially infecting loved ones); and stress (due to longer working hours and increased workload). Conclusion: COVID-19 has affected the mental health of HCWs. We identified a number of factors which may be contributing to a deterioration in mental health for participants from diverse ethnic backgrounds. Healthcare organisations should consider developing strategies to counter the negative impact of these factors, including recommendations made by HCWs themselves.

Effects of Lactobacillus plantarum P9 Probiotics on Defecation and Quality of Life of Individuals with Chronic Constipation: Protocol for a Randomized, Double-Blind, Placebo-Controlled Clinical Trial.

Background Although probiotics have been shown to improve constipation-related symptoms, a clear consensus on the use of probiotics as a constipation-relieving agent has not been reached, which is attributed to the limited available evidence and inconsistent protocols used in existing studies. Method A randomized, double-blind, placebo-controlled clinical trial is designed to study the efficiency and possible mechanism of action of probiotics for chronic constipation, in which 200 eligible volunteers with chronic constipation will be randomly assigned to a probiotic group (oral Lactobacillus plantarum P9 probiotic powder, 100 billion colony-forming units (CFUs)/day) or a placebo group. Volunteers, treatment distributors, data collectors, and data analysts will be blinded. The primary outcome is the weekly mean frequency of complete spontaneous bowel movements (CSBMs), and secondary outcomes include weekly mean frequency of CSBMs ≥3, weekly mean frequency of spontaneous bowel movements (SBMs), weekly mean stool appearance score, weekly mean difficulty of passing stool score, weekly percentage of volunteers who use auxiliary measures to assist with defecation (WPUAMA), quality-of-life (QOL) score, emotional status score, gut microbiome, and faecal metabolome. Each outcome measure will be assessed at the time points of preadministration (day 0), administration (day 14 and/or 28), and postadministration (day 42) to identify inter- and intragroup differences. Adverse events will be recorded to evaluate the safety of L. plantarum P9. Discussion. The protocol will provide methodological guidance for other similar studies, avoiding methodological bias and ultimately facilitating the formulation of consensus on the use of probiotics as a constipation-relieving agent. In addition, the results are more comprehensive than those of existing studies and may objectively and scientifically reflect the effectiveness of L. plantarum P9 on constipation. If the expected study findings are obtained, L. plantarum P9, taken as a probiotic, may become a complementary choice for chronically constipated patients. This trial is registered with Chinese Clinical Trial Registry (ChiCTR) (no. ChiCTR2000038396) registered on November 22, 2020, https://www.chictr.org.cn/showproj.aspx?proj=54024.

A Study on Information Classification and Storage in Cloud Computing Data Centers Based on Group Collaborative Intelligent Clustering

Internet of things (IoT) and cloud computing are combined to form a cloud computing data center, and cloud computing provides virtualization, storage, computing, and other support services for IoT applications. Data is the foundation and core of cloud IoT platform applications, and massive multisource heterogeneous IoT data aggregation and storage have basic requirements such as real-time, security, and scalability. This paper focuses on the aggregation and storage methods of massive heterogeneous cloud IoT data, solving the multisource data aggregation problem caused by inconsistent protocols and the heterogeneous data storage problem caused by inconsistent data types. A heterogeneous network protocol adaptation and data aggregation method is proposed for the multisource data aggregation problem caused by protocol inconsistency. A protocol adaptation layer is set up in the IoT virtual gateway to achieve compatibility with multiple types of data aggregation protocols, ensuring adaptive access to different types of IoT nodes, and on this basis, data is transmitted to the cloud IoT platform through a unified interface, shielding the variability of IoT sensing devices. Given the problems of device forgery and malicious tampering in the data aggregation process, we propose a fast authentication and data storage method for IoT devices based on “API key” and implant the device authentication API key into the protocol adaptation layer of the virtual gateway to realize the source authentication of IoT nodes and ensure the authenticity of data.

Microengineered filters for efficient delivery of nanomaterials into mammalian cells

Intracellular delivery of nanomaterials into the cells of interest has enabled cell manipulation for numerous applications ranging from cell-based therapies to biomedical research. To date, different carriers or membrane poration-based techniques have been developed to load nanomaterials to the cell interior. These biotools have shown promise to surpass the membrane barrier and provide access to the intracellular space followed by passive diffusion of exogenous cargoes. However, most of them suffer from inconsistent delivery, cytotoxicity, and expensive protocols, somewhat limiting their utility in a variety of delivery applications. Here, by leveraging the benefits of microengineered porous membranes with a suitable porosity, we demonstrated an efficient intracellular loading of diverse nanomaterials to different cell types based on inducing mechanical disruption to the cell membrane. In this work, for the first time, we used ultra-thin silicon nitride (SiN) filter membranes with uniform micropores smaller than the cell diameter to load impermeable nanomaterials into adherent and non-adherent cell types. The delivery performance using SiN microsieves has been validated through the loading of functional nanomaterials from a few nanometers to hundreds of nanometers into mammalian cells with minimal undesired impacts. Besides the high delivery efficiency and improved cell viability, this simple and low-cost approach offers less clogging and higher throughput (107 cell min−1). Therefore, it yields to the efficient introduction of exogenous nanomaterials into the large population of cells, illustrating the potential of these microengineered filters to be widely used in the microfiltroporation (MFP) setup.

Use of hyperbaric oxygen therapy for preventing delayed neurological sequelae in patients with carbon monoxide poisoning: A multicenter, prospective, observational study in Japan.

The purpose of this study was to clarify the practical clinical treatment for acute carbon monoxide (CO) poisoning in Japan and to investigate the efficacy of hyperbaric oxygen (HBO2) therapy in preventing delayed neurological sequelae (DNS) in the acute phase of CO poisoning. We conducted a multicenter, prospective, observational study of acute CO poisoning in Japan. Patients with acute CO poisoning were enrolled and their treatment details were recorded. The primary endpoint was the onset of DNS within 2 months of CO exposure. Factors associated with DNS were assessed with logistic regression analysis. A total of 311 patients from 57 institutions were registered and 255 were analyzed: 171 received HBO2 therapy (HBO2 group) and 84 did not (normobaric oxygen [NBO2] group). HBO2 therapy was performed zero, once, twice, or three times within the first 24 h in 1.8%, 55.9%, 30.9%, and 11.3% of the HBO2 group, respectively. The treatment pressure in the first HBO2 session was 2.8 ATA (47.9% of the HBO2 group), 2.0 ATA (41.8%), 2.5 ATA (7.9%), or another pressure (2.4%). The incidence of DNS was 13/171 (7.6%) in the HBO2 group and 3/84 (3.6%) in the NBO2 group (P = 0.212). The number of HBO2 sessions in the first 24 h was one of the factors associated with the incidence of DNS (odds ratio, 2.082; 95% confidence interval, 1.101-3.937; P = 0.024). The practical clinical treatment for acute CO poisoning, including HBO2 therapy, varied among the institutions participating in Japan. HBO2 therapy with inconsistent protocols showed no advantage over NBO2 therapy in preventing DNS. Multiple HBO2 sessions was associated with the incidence of DNS.

Perceived barriers to the adoption of active surveillance in low-risk prostate cancer: a qualitative analysis of community and academic urologists

BackgroundClinical practice guidelines recommend active surveillance as the preferred treatment option for low-risk prostate cancer, but only a minority of eligible men receive active surveillance, and practice variation is substantial. The aim of this study is to describe barriers to urologists’ recommendation of active surveillance in low-risk prostate cancer and explore variation of barriers by setting.MethodsWe conducted semi-structured interviews among 22 practicing urologists, evenly distributed between academic and community practice. We coded barriers to active surveillance according to a conceptual model of determinants of treatment quality to identify potential opportunities for intervention.ResultsCommunity and academic urologists were generally in agreement on factors influencing active surveillance. Urologists perceived patient-level factors to have the greatest influence on recommendations, particularly tumor pathology, patient age, and judgements about the patient’s ability to adhere to follow-up protocols. They also noted cross-cutting clinical barriers, including concerns about the adequacy of biopsy samples, inconsistent protocols to guide active surveillance, and side effects of biopsy procedures. Urologists had differing opinions on the impact of environmental factors, such as financial disincentives and fear of litigation.ConclusionsDespite national and international recommendations, both academic and community urologists note a variety of barriers to implementing active surveillance in low risk prostate cancer. These barriers will need to be specifically addressed in efforts to help urologists offer active surveillance more consistently.

Transplanted Erythropoietin-Expressing Mesenchymal Stem Cells Promote Pro-survival Gene Expression and Protect Photoreceptors From Sodium Iodate-Induced Cytotoxicity in a Retinal Degeneration Model.

Mesenchymal stem cells (MSC) are highly regarded as a potential treatment for retinal degenerative disorders like retinitis pigmentosa and age-related macular degeneration. However, donor cell heterogeneity and inconsistent protocols for transplantation have led to varied outcomes in clinical trials. We previously showed that genetically-modifying MSCs to express erythropoietin (MSCEPO) improved its regenerative capabilities in vitro. Hence, in this study, we sought to prove its potential in vivo by transplanting MSCsEPO in a rat retinal degeneration model and analyzing its retinal transcriptome using RNA-Seq. Firstly, MSCsEPO were cultured and expanded before being intravitreally transplanted into the sodium iodate-induced model. After the procedure, electroretinography (ERG) was performed bi-weekly for 30 days. Histological analyses were performed after the ERG assessment. The retina was then harvested for RNA extraction. After mRNA-enrichment and library preparation, paired-end RNA-Seq was performed. Salmon and DESeq2 were used to process the output files. The generated dataset was then analyzed using over-representation (ORA), functional enrichment (GSEA), and pathway topology analysis tools (SPIA) to identify enrichment of key pathways in the experimental groups. The results showed that the MSCEPO-treated group had detectable ERG waves (P <0.05), which were indicative of successful phototransduction. The stem cells were also successfully detected by immunohistochemistry 30 days after intravitreal transplantation. An initial over-representation analysis revealed a snapshot of immune-related pathways in all the groups but was mainly overexpressed in the MSC group. A subsequent GSEA and SPIA analysis later revealed enrichment in a large number of biological processes including phototransduction, regeneration, and cell death (Padj <0.05). Based on these pathways, a set of pro-survival gene expressions were extracted and tabulated. This study provided an in-depth transcriptomic analysis on the MSCEPO-treated retinal degeneration model as well as a profile of pro-survival genes that can be used as candidates for further genetic enhancement studies on stem cells.

Concerns About Mesenchymal Stem Cell Therapy

Dear Editor, Multipotent mesenchymal stromal cells (MSCs), because of their potential for treating diseases, have gained considerable attention over the past 30 years for their immunosuppressive properties and tissue regeneration capabilities.1,2 This grew into an interest in the 1970s by Friedenstein and contemporaries after studying the bone m ar row.3 MSCs are an excellent candidate for cell therapy because of having intrinsic differentiation potentials into bone, cartilage and fat cells not found previously in other cells. They can also be isolated and expanded easily in vitro, and produce abundant useful growth factors and cytokines.2,4Mesenchymal stem cells have become the top used stem cell type for clinical application with encouraging results.5,6 Significant progress has been made in stem cell research in recent years. The main therapeutic effects of MSCs are now attributed to the stimulation of several innate repair processes in injured tissues in vivo by secreted factors as well as the immunomodulation response. Therefore, MSC therapy is expected to find clinical application in human diseases.6,7 However, there are some critical issues that need to be addressed before MSCs can be used for clinical therapy in humans which will help determine the efficiency of cells administered to the patients as a therapeutic approach, most important of which is immunosuppressive properties.8,9 MSCs can be affected by tumor support that may affect potential tumorigenesis after MSCs transplantation, including different donors and tissues, inconsistent protocols, varying dosages and differing transfusion patterns, and mechanisms that control the behavior of the MSCs at the target site.10,11 Because of their unique features, stem cells are undoubtedly a great hope for the treatment of many diseases. More research is developing on the potential long-term risks associated with MSC therapy. However, Additional studies would also be a major contribution to stem cell biology in general as well as their transplantation.

An automated, high-throughput methodology optimized for quantitative cell-free mitochondrial and nuclear DNA isolation from plasma

Progress in the study of circulating, cell-free nuclear DNA (ccf-nDNA) in cancer detection has led to the development of noninvasive clinical diagnostic tests and has accelerated the evaluation of ccf-nDNA abundance as a disease biomarker. Likewise, circulating, cell-free mitochondrial DNA (ccf-mtDNA) is under similar investigation. However, optimal ccf-mtDNA isolation parameters have not been established, and inconsistent protocols for ccf-nDNA collection, storage, and analysis have hindered its clinical utility. Until now, no studies have established a method for high-throughput isolation that considers both ccf-nDNA and ccf-mtDNA. We initially optimized human plasma digestion and extraction conditions for maximal recovery of these DNAs using a magnetic bead-based isolation method. However, when we incorporated this method onto a high-throughput platform, initial experiments found that DNA isolated from identical human plasma samples displayed plate edge effects resulting in low ccf-mtDNA reproducibility, whereas ccf-nDNA was less affected. Therefore, we developed a detailed protocol optimized for both ccf-mtDNA and ccf-nDNA recovery that uses a magnetic bead-based isolation process on an automated 96-well platform. Overall, we calculate an improved efficiency of recovery of ∼95-fold for ccf-mtDNA and 20-fold for ccf-nDNA when compared with the initial procedure. Digestion conditions, liquid-handling characteristics, and magnetic particle processor programming all contributed to increased recovery without detectable positional effects. To our knowledge, this is the first high-throughput approach optimized for ccf-mtDNA and ccf-nDNA recovery and serves as an important starting point for clinical studies.

Evidence for contamination as the origin for bacteria found in human placenta rather than a microbiota.

Until recently the in utero environment of pregnant women was considered sterile. Recent high-sensitivity molecular techniques and high-throughput sequencing lead to some evidence for a low-biomass microbiome associated with the healthy placenta. Other studies failed to reveal evidence for a consistent presence of bacteria using either culture or molecular based techniques. Comparing conflicting “placental microbiome” studies is complicated by the use of varied and inconsistent protocols. Given this situation, we undertook an evaluation of the in utero environment sterility using several controlled methods, in the same study, to evaluate the presence or absence of bacteria and to explain contradictions present in the literature. Healthy pregnant women (n = 38) were recruited in three maternity wards. Placenta were collected after cesarean section with or without Alexis® and vaginal delivery births. For this study we sampled fetal membranes, umbilical cord and chorionic villi. Bacterial presence was analyzed using bacterial culture and qPCR on 34 fetal membranes, umbilical cord and chorionic villi samples. Shotgun metagenomics was performed on seven chorionic villi samples. We showed that the isolation of meaningful quantities of viable bacteria or bacterial DNA was possible only outside the placenta (fetal membranes and umbilical cords) highlighting the importance of sampling methods in studying the in utero environment. Bacterial communities described by metagenomics analysis were similar in chorionic villi samples and in negative controls and were dependent on the database chosen for the analysis. We conclude that the placenta does not harbor a specific, consistent and functional microbiota.

The effect of theta-burst stimulation on unilateral spatial neglect following stroke: a systematic review

Objective: To evaluate the effectiveness of theta-burst stimulation for the treatment of stroke-induced unilateral spatial neglect. Data sources: A systematic literature search was conducted from the inception of each database to 30 June 2018 using CINAHL, EMBASE, PubMed, PsycINFO, and Scopus. Review methods: Articles were included if theta-burst stimulation was used to treat neglect following a stroke. The additional a priori inclusion criteria were as follows: (1) ⩾3 adult (⩾18 years) participants, (2) ⩾50% stroke population, and (3) peer-reviewed journal articles published in English. Extracted data included study and treatment characteristics, results, and adverse events. Results: Nine studies met the inclusion criteria, generating a total of 148 participants. Eight studies evaluated a continuous stimulation protocol and one study investigated an intermittent stimulation protocol. Overall, both protocols significantly improved neglect severity when compared against placebo or active controls (P < 0.05). Adding smooth pursuit training to theta-burst stimulation did not improve neglect relative to when the stimulation was delivered alone (P > 0.05). There was inconsistent reporting of neglect terminology, outcome measures, and adverse events. The treatment characteristics were heterogeneous among the trials. Conclusion: This systematic review found that theta-burst stimulation seems to improve post-stroke unilateral spatial neglect, but because the evidence is limited to a few small studies with varied and inconsistent protocols and use of terminology, no firm conclusion on effectiveness can be drawn.