A Prevention Protocol Reduces Spinal Cord Ischemia in Patients Undergoing Branched/Fenestrated Endovascular Aortic Repair.

OBJECTIVES: Spinal cord ischemia (SCI) is a devastating complication that is associated with thoracoabdominal aortic repair, with higher risk associated with increased aortic coverage length, making patients undergoing branched/fenestrated endovascular repair(B/FEVAR) particularly vulnerable. A bundled SCI prevention protocol was previously reported to reduce SCI rates when compared to a historic cohort in a single-center study. Therefore, this analysis aims to further validate and update outcomes associated with the protocol given the routine implementation of this strategy at two institutions (University of Florida [UF] and the University of Alabama at Birmingham [UAB]) since inception. METHODS: Components of the SCI prevention protocol include selective cerebrospinal fluid (CSF) drainage, specified blood pressure parameters, transfusion goals, and selective pharmacologic adjuncts (naloxone, steroids). This protocol was routinely implemented in May 2015. Patients undergoing B/FEVAR from May 2015-December 2022 constituted the post-protocol cohort(n=402) and were compared to the pre-protocol cohort (n=160, January 2010-April 2015). The primary outcome was SCI incidence and subgroup analysis was conducted among patients deemed to be high-risk (Crawford extent I- III thoracoabdominal aneurysms (TAAA) dissection-related disease, prior aortic repair, coverage proximal to zone 5). Survival analysis was performed using Kaplan-Meier methodology. RESULTS: The pre- and post-protocol cohorts were demographically similar, though more post-protocol patients were American Society of Anesthesiology(ASA) class IV (86.1% vs. 55.0%; p<0.001). TAAA was the most common indication in both groups. CSF drain placement was more common in the post-protocol group, particularly among high-risk patients. SCI occurred in 15.9% of pre-protocol patients versus 3.0% of post-protocol patients(p<0.001). In high-risk patients, the pre- and post-protocol cohort SCI incidence was 23.2% vs. 5.0%, respectively (p<0.001). 30-day mortality was decreased in the post-protocol cohort (6.3% vs. 2.2%, p=0.02). Although the post-protocol group had a trend toward improved 1-year survival, this was not statistically significant (84.4% vs. 88.3%, log-rank p=0.35). Among SCI patients, one-year mortality was 28% and 33.3% in the pre- and post-protocol groups, respectively(p=0.46). CONCLUSION: Implementation of a bundled SCI prevention protocol significantly reduces SCI rates in B/FEVAR patients, which has now been validated at two institutions, with the most significant reductions occurring among high-risk patients. Although the overall one-year mortality difference was not significantly different between the cohorts, the high mortality rates among SCI patients highlights the importance of preventative measures.

Exploring the Potential of In vitro Maturation (IVM) of Oocytes: Indications, Applications, and Treatment Protocols.

This review addresses the current understanding of In Vitro Maturation (IVM) treatment, including indications and effective treatment protocols influencing oocyte developmental competence. A comprehensive literature search was performed to gather relevant studies, clinical trials, and reviews related to IVM. Databases such as PubMed, MEDLINE, and pertinent medical journals were searched. The selected literature was analyzed and synthesized to offer a comprehensive overview. IVM has emerged as a promising technique for inducing maturation in immature oocytes across various developmental stages. Its applications extend to areas utilizing In Vitro Fertilization (IVF), gaining traction as a treatment option for Polycystic Ovary Syndrome (PCOS) and fertility preservation in cancer patients. Recent advancements have led to improved global pregnancy rates, resulting in successful births. IVM also holds potential in reducing risks associated with conventional IVF, including ovarian hyperstimulation syndrome and multiple pregnancies. Despite these advantages, IVM adoption in clinical practice remains limited. Ongoing research aims to refine therapeutic protocols and expand clinical indications. IVM holds promise in assisted reproductive technology, spanning applications from cancer patient fertility preservation to addressing PCOS. Enhanced pregnancy rates highlight efficacy, while risk reduction compared to IVF underscores its importance. Further research is needed for optimal use across patient groups, emphasizing protocol refinement and expanded applications.

Protocol for enriching, isolating, and testing drug susceptibility of facultative anaerobic fungi.

Research on fungi under anaerobic conditions is limited but crucial for understanding their ecological and pathological impacts. Here, we present a protocol for enriching, isolating, and characterizing anaerobic fungi from environmental and clinical samples. We also describe steps for evaluating the anaerobic growth potential and drug susceptibility of fungal pathogens. This protocol can contribute to the need for initiating effective antifungal therapy to address and manage fungal infections or mycosis in oxygen-limited environments. For complete details on the use and execution of this protocol, please refer to Yadav et al.1.

Clinical Difficulties Related to Direct Composite Restorations: A Multinational Survey.

AIMS: Composite materials are widely used in dentistry for direct tooth restorations. However, they are highly sensitive to the working technique employed during the restorative procedure. Even minor procedural errors can have a significant impact on the quality including the longevity of the restoration. Hence the aim of this study was to determine the material preferences and analyse the clinical problems associated with direct composite restorations in a cohort of dentists. METHODS: A 20-item online questionnaire was created in English and administered 1830 general dentists and specialists in 13 countries. The first section of the questionnaire included four questions to elicit demographic data, and the second section comprised 16 questions focused on material preferences for conservative restorations, durability of composite restorations, and the most challenging stages the dentists faced during the composite restorative procedures. RESULTS: Respondents decided most often to use composite materials for the tooth restorations (OR 997.4, 95% CI 233.8-4254.8, P value <.001). Most respondents indicated that the durability of composite restorations was approximately 7 to 10 years (41.5%). Among the factors affecting durability, maintenance of a dry cavity was the most often reported reason (47.1%) and the foremost challenge faced by dentists (61.0%) during the composite restorative procedures. CONCLUSIONS: Our study confirmed that resin-based composites are the most popular material for direct restoration in many countries. Although working with this material is difficult and involves multiple steps, maintaining a dry cavity during bonding, and material application may affect the therapeutic success and durability of these restorations. Clinicians need to be attentive to this issue and be prepared to adapt their decision-making and consider opting for alternative restorative materials, if appropriate.

Respiratory Therapist-Driven Mechanical Ventilation Protocol Is Associated With Increased Lung Protective Ventilation.

BACKGROUND: Mechanical ventilation is a common life-saving procedure but can lead to serious complications, including ARDS and oxygen toxicity. Nonadherence to lung-protective ventilation guidelines is common. We hypothesized that a respiratory therapist-driven mechanical ventilation bundle could increase adherence to lung-protective ventilation and decrease the incidence of pulmonary complications in the ICU. METHODS: A respiratory therapist-driven protocol was implemented on August 1, 2018, in all adult ICUs of a Midwestern academic tertiary center. The protocol targeted low tidal volume, adequate PEEP, limiting oxygen, adequate breathing frequency, and head of the bed elevation. Adherence to lung-protective guidelines and clinical outcomes were retrospectively observed in adult subjects admitted to the ICU and on ventilation for ≥ 24 h between January 2011 and December 2019. RESULTS: We included 666 subjects; 68.5% were in the pre-intervention group and 31.5% were in the post-intervention group. After adjusting for body mass index and intubation indication, a significant increase in overall adherence to lung-protective ventilation guidelines was observed in the post-intervention period (adjusted odds ratio 2.48, 95% CI 1.73-3.56). Fewer subjects were diagnosed with ARDS in the post-intervention group (adjusted odds ratio 0.22, 95% CI 0.08-0.65) than in the pre-intervention group. There was no difference in the incidence of ventilator-associated pneumonia, ventilator-free days, ICU mortality, or death within 1 month of ICU discharge. CONCLUSIONS: A respiratory therapist-driven protocol increased adherence to lung-protective mechanical ventilation guidelines in the ICU and was associated with decreased ARDS incidence.

A protocol to quantify cross-sectional and longitudinal differences in duction patterns.

Currently, there is no established system for quantifying patterns of ocular ductions. This poses challenges in tracking the onset and evolution of ocular motility disorders, as current clinical methodologies rely on subjective observations of individual movements. We propose a protocol that integrates image processing, a statistical framework of summary indices, and criteria for evaluating both cross-sectional and longitudinal differences in ductions to address this methodological gap. We demonstrate that our protocol reliably transforms objective estimates of ocular rotations into normative patterns of total movement area and movement symmetry. This is a critical step towards clinical application in which our protocol could first diagnose and then track the progression and resolution of ocular motility disorders over time.

Protocol for the creation and characterization of SARS-CoV-2 variant testing panels using remnant clinical samples for diagnostic assay testing.

The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Alpha variant in 2020 demonstrated the need for reanalysis of diagnostic tests to ensure detection of emerging variants. Here, we present a protocol for creating and characterizing SARS-CoV-2 variant testing panels using remnant clinical samples for diagnostic assay testing. We describe steps for characterizing SARS-CoV-2 remnant clinical samples and preparing them into pools and their use in preparing varying quantities of virus. We then detail procedures for verifying variant detection using the resulting sample panel. For complete details on the use and execution of this protocol, please refer to Rao et al.1,2.

Protocol for producing hyperpolarized 13C-bicarbonate for clinical MRI of extracellular pH in aggressive tumors.

Tumor acidosis is one of the hallmarks indicating the initiation and progression of various cancers. Here, we present a protocol for preparing a hyperpolarized (HP) 13C-bicarbonate tissue pH MRI imaging contrast agent to detect aggressive tumors. We describe the steps for the formulation and polarization of a precursor molecule 13C-glycerol carbonate (13C-GLC), the post-dissolution reaction, and converting HP 13C-GLC to an injectable HP 13C-bicarbonate solution. We then detail procedures for MRI data acquisition to generate tumor pH maps for assessing tumor aggressiveness. For complete details on the use and execution of this protocol, please refer to Mu et al.1.

Protocol to develop a chemotherapy drug screening process by constructing a cancer prognostic model using public databases.

Drug resistance is currently the biggest challenge in cancer chemotherapy. Here, we present a protocol to develop a chemotherapy drug screening process by constructing a cancer prognostic model (PM) using public databases. We describe steps for downloading code and data, preparing the expression matrix and metadata for analysis, screening modeling genes, and constructing a PM. We then detail procedures for constructing predictive websites for cancer patients' survival based on their age, tumor stage, gene expression levels, and risk scores. For complete details on the use and execution of this protocol, please refer to Bai et al.1.

Protocol for phage matching, treatment, and monitoring for compassionate bacteriophage use in non-resolving infections.

Phage therapy has re-emerged as a promising treatment for non-resolving infections. Given the lack of approved phage treatments, there is a need to establish a compassionate use pipeline. Here, we present a protocol for phage matching, treatment, and monitoring for compassionate bacteriophage use in non-resolving infections. We describe steps for consultation and request implementation, evaluating and comparing different aspects of phage activity, and phage production. We then detail procedures for multidisciplinary meetings, ethics approvals, phage therapy, and follow-up. For complete details on the use and execution of this protocol, please refer to Onallah et al.1,2.

Protocol for deep mining the correlation between acute pancreatitis and ferroptosis using the MIMIC-III database and STATA software.

The limitations associated with distinguishing serum Fe2+ and Fe3+ hinder the widespread application of ferroptosis, beyond laboratory settings. Here, we present a protocol for deep mining the correlation between acute pancreatitis and ferroptosis using the MIMIC-III database and STATA software. We describe steps for using Cox regression, decision curve analysis (DCA), and receiver operating characteristic (ROC) approaches to establish the relationship between them and determine the relevant factors. This protocol has potential application in establishing novel research models that integrate both fundamental and clinical methodologies. For complete details on the use and execution of this protocol, please refer to Yueling Deng et al.1.

Protocol for extracting and isolating porcine bone-marrow-derived macrophages from ribs.

Due to anatomical and biological similarities with humans, pigs are increasingly used for inflammation- and immune-related studies in biomedical research, including the field of osteonecrosis and osteoimmunology. Here, we present a protocol for rib extraction, isolation of the bone marrow by centrifugation, and processing to obtain bone-marrow-derived macrophages (BMDMs). Then, we describe the procedures of in vitro experiments to evaluate the cell phenotype. For complete details on the use and execution of this protocol, please refer to Andre et al.1.

The distance of insertion points from wound margins in interrupted and vertical mattress sutures influences the tensile load capacity: An in vitro experimental study.

AIM: To determine the tensile load capacity (TLC) and the tearing characteristics for interrupted and vertical mattress sutures with different insertion points from the wound margin, and the effect of the bite size when using vertical mattress sutures. MATERIALS AND METHODS: A total of 120 gingiva and lining mucosa samples obtained from pig jaws were divided into groups according to the suturing technique (interrupted and vertical mattress sutures), distance of the insertion points from the wound margin (margin, 1, 3, and 5 mm) and bite size (1, 3, and 5 mm). The TLC of the suture and the tearing characteristics were evaluated using a tensile tester device. RESULTS: The TLC was significantly higher for vertical mattress sutures than for interrupted sutures regardless of the distance of the insertion points from the wound margin (intergroup p < .001). This distance significantly influenced the TLC for vertical mattress sutures (p < .05) but not for interrupted sutures (p > .05). Testing the tearing characteristics revealed that no tissue tearing occurred in groups when the insertion points were more than 3 mm from the wound margin. CONCLUSION: The TLC is higher for vertical mattress sutures than for interrupted sutures, and it increases when the insertion points are farther from the wound margin.

Protocol to identify biomarkers in patients with post-COVID condition using multi-omics and machine learning analysis of human plasma.

Here, we present a workflow for analyzing multi-omics data of plasma samples in patients with post-COVID condition (PCC). Applicable to various diseases, we outline steps for data preprocessing and integrating diverse assay datasets. Then, we detail statistical analysis to unveil plasma profile changes and identify biomarker-clinical variable associations. The last two steps discuss machine learning techniques for unsupervised clustering of patients based on their inherent molecular similarities and feature selection to identify predictive biomarkers. For complete details on the use and execution of this protocol, please refer to Wang et al.1.

Modified Gwakjeongtang for Diarrhea-Predominant Irritable Bowel Syndrome: Study Protocol for a Randomized, Double-Blind, Placebo-Controlled, Pilot Clinical Trial.

Introduction: Irritable bowel syndrome (IBS) is a chronic condition characterized by recurrent abdominal pain associated with bowel movements. Modified Gwakjeongtang (MGT), an herbal prescription rooted in traditional East Asian medicine, consists of thirteen botanical drugs known for their potential to enhance intestinal barrier function, regulate gastrointestinal motility, and exhibit anti-inflammatory and antioxidant properties. Despite a few previous clinical trials highlighting MGT's potential for IBS symptom management, limited evidence exists with placebo control. Methods and Analysis: In this pilot randomized clinical trial protocol, we aim to exploratively evaluate the efficacy and safety of MGT in patients with diarrhea-predominant IBS (IBS-D) by comparing it with a placebo. A total of 60 IBS-D patients will be enrolled, and eligible participants will be randomly allocated to either the MGT or placebo groups. Over a 4-week period, they will receive MGT or placebo granules three times a day. The primary endpoint will be the overall response rate post-treatment, determined through daily assessments of abdominal pain intensity and stool consistency. Ethics and Dissemination: This clinical trial protocol has received approval from the Korean Ministry of Food and Drug Safety for an investigational new drug application and Institutional Review Board of the Kyung Hee University Korean Medicine Hospital. The research findings will be submitted and published in international peer-reviewed journal. Trial Registration: Clinical research information service (registration number: KCT0008523).

Protocol for identifying metabolite biomarkers in patient uterine fluid for early ovarian cancer detection.

High mortality of ovarian cancer (OC) is primarily attributed to the lack of effective early detection methods. Uterine fluid, pooling molecules from neighboring ovaries, presents an organ-specific advantage over conventional blood samples. Here, we present a protocol for identifying metabolite biomarkers in uterine fluid for early OC detection. We describe steps for uterine fluid collection from patients, metabolite extraction, metabolomics experiments, and candidate metabolite biomarker screening. This standardized workflow holds the potential to achieve early OC diagnosis in clinical practice. For complete details on the use and execution of this protocol, please refer to Wang et al.1.

Protocol for predicting multivariate change of brain patterns using model-informed fMRI activations.

Investigating the spatially distributed information contained in fMRI data is essential for understanding brain functions. Here, we present a protocol to dynamically predict short-term changes in neural patterns using trial-by-trial blood-oxygen-level-dependent (BOLD) activity of a seed region. We describe steps for setting fMRI data acquisition parameters and quantification of changes in multivariate patterns. We then detail procedures for defining seed regions and identifying brain areas in which changes in multivariate patterns can be predicted by BOLD activity of the seed region. For complete details on the use and execution of this protocol, please refer to Möhring et al.1.

Protocol to measure apoptosis-associated speck-like protein containing a CARD specks in human cerebrospinal fluid via imaging flow cytometry.

Apoptosis-associated speck-like protein containing a c-terminal caspase activation and recruitment domain (ASC) specks are elevated in the cerebrospinal fluid (CSF) of Alzheimer's disease and related dementias (AD/ADRDs) patients. Here, we present a flow cytometry protocol to quantify ASC specks. We describe steps for fluorescently labeling ASC specks using antibody technology, visualizing with imaging flow cytometry, and gating based on physical characteristics. CSF ASC specks levels positively correlate with phosphorylated tau (Thr181) and negatively correlate with amyloid ß ratio (42/40), thus serving as a neuroinflammatory biomarker for diagnosing AD/ADRDs. For complete details on the use and execution of this protocol, please refer to Jiang et al.1.

Development of an inpatient protocol for adolescents with avoidant/restrictive food intake disorder: a case study.

Introduction: Avoidant/restrictive food intake disorder (ARFID) is an eating disorder characterised by a pattern of eating that leads to failure to meet appropriate nutritional and/or energy needs. Method: In the absence of evidence-based inpatient guidelines for adolescents with ARFID, we set out to develop and pilot an inpatient protocol for adolescents with ARFID. Identification of the key differences between managing inpatients with ARFID and anorexia nervosa (AN) led to modification of an existing AN protocol with the goals of better meeting patient needs, enhancing alignment with outpatient care, and improving outcomes. A case report of an adolescent with ARFID who had three hospital admissions is presented to highlight these changes. Interviews with this patient and her family were undertaken, together with key staff, to explore the challenges of the AN protocol for this patient and the perceived benefits and any limitations of the ARFID protocol for this patient and others. Results: The new ARFID protocol supports greater choice of meals, without the need for rest periods after meals and bathroom supervision. The similarities with the AN protocol reflect the need to promote timely weight gain through meal support, including a staged approach to nutritional supplementation. The protocol appears to have been well accepted by the patient and her family, as well as by staff, and continues to be used in cases of ARFID. Conclusion: Further evaluation would help identify how well this protocol meets the needs of different adolescents with ARFID.

Introduction: Le trouble évitant/restrictif de la prise alimentaire (TERPA) est un trouble alimentaire caractérisé par un modèle d'alimentation qui entraîne une incapacité à répondre aux besoins nutritionnels et/ou énergétiques appropriés. Méthode: En l'absence de lignes directrices fondées sur des données probantes en milieu hospitalier pour des adolescents souffrant de TERPA, nous avons entrepris de développer et de piloter un protocole en milieu hospitalier pour les adolescents souffrant de TERPA. L'identification des principales différences entre la prise en charge des patients hospitalisés souffrant de TERPA et d'anorexie mentale (AM) a mené à une modification d'un protocole d'AM existant dans le but de mieux répondre aux besoins des patients, d'accroître l'alignement avec les soins des patients ambulatoires, et d'améliorer les résultats. Un rapport de cas d'une adolescente souffrant de TERPA qui a eu trois hospitalisations est présenté pour souligner ces changements. Des entrevues avec cette patiente et sa famille ont été réalisées, de même qu'avec le personnel principal, afin d'explorer les difficultés du protocole d'AM pour cette patiente ainsi que les avantages perçus et toute limite du protocole TERPA pour cette patiente et d'autres. Résultats: Le nouveau protocole TERPA supporte un plus grand nombre de repas, sans le besoin de périodes de repos après les repas et une supervision de la salle de bain. Les similitudes avec le protocole AM reflètent le besoin de promouvoir une prise de poids rapide grâce à un soutien aux repas, y compris une approche par étapes de supplémentation nutritionnelle. Le protocole semble avoir été bien accepté par la patiente et sa famille, ainsi que par le personnel, et continue d'être utilisé dans les cas de TERPA. Conclusion: Une évaluation plus poussée aiderait à identifier dans quelle mesure ce protocole répond aux besoins de différents adolescents souffrant de TERPA.

Comparison In Vitro Fertilization Outcomes between DouStim and Minimal Stimulation Protocols in Poor Ovarian Responders: A Randomized Clinical Trial.

BACKGROUND: Various protocols have been approved to improve the response rate leading to successful fertilization in poor ovarian responders (PORs). The application of double ovarian stimulation (DuoStim) in the follicular and luteal phases of the same ovarian cycle has been shown as an intriguing option to achieve more oocyte retrievals in the shortest time. The aim of the current study is to compare the outcomes of different protocols, minimal stimulation (MS) and Duostim. MATERIALS AND METHODS: This randomized clinical trial was performed on 42 in vitro fertilization (IVF) candidates with POR diagnosis. Patients were classified into two equal groups and treated with the DuoStim protocol and MS protocol. The IVF outcomes, including retrieved follicles, oocytes, metaphase II (MII) oocytes and embryos, were compared between these groups. RESULTS: The patients' characteristics including age, anti-mullerian hormone (AMH), follicle stimulating hormone (FSH), luteinizing hormone (LH), and antral follicle count (AFC) were collected and compared. It showed there was no significant difference between the two groups baseline characteristics (P>0.05). We observed that the DuoStim protocol resulted in a significantly higher score in comparison with the MS protocols , including the number of follicles (6.23 ± 2.93 vs. 1.77 ± 1.66, P<0.001), retrieved oocytes (3.86 ± 2.57 vs. 1.68 ± 1.58, P=0.002), MII oocytes (3.36 ± 2.42 vs. 1.27 ± 1.27, P=0.001) and obtained embryos (2.04 ± 1.64 vs. 0.77 ± 0.86, P=0.003). CONCLUSION: The DuoStim protocol is a favourable and time saving plan that is associated with more oocytes in a single stimulation cycle. The DuoStim protocol significantly can result in more frequent MII oocytes and embryos. We figured that the higher number of oocytes and embryos might have led to a higher rate of pregnancy (registration number: IRCT20200804048303N1).