Concentration of voxelotor in sickle cell disease can be estimated using electrophoresis and high-performance liquid chromatography.

OBJECTIVES: Voxelotor can increase hemoglobin levels in patients living with sickle cell disease (SCD). A clinician who is monitoring voxelotor response may want to know whole-blood voxelotor concentration, but this cannot be measured in most clinical settings. However, voxelotor has been demonstrated to cause "peak splitting" in common methods of hemoglobin measurement such as capillary zone electrophoresis (CZE) and high-performance liquid chromatography (HPLC). We hypothesized that we could use the size of the peak split to estimate the whole-blood concentration. METHODS: Blood from people with SCD was dosed with known concentrations of voxelotor, and multiparameter regression was used to derive the relationship of voxelotor concentration to the degree of peak splitting observed. To validate these equations, 21 patients started on voxelotor at 1500 mg/d had blood samples drawn at days 0, 14, 30, and 60. Samples were sent out for gold standard voxelotor concentration testing. The derived equations were then used to calculate voxelotor concentration. RESULTS: Calculated concentrations correlated strongly with measured concentrations for both CZE (R2 = 0.83, P < .001) and HPLC (R2 = 0.76, P < .001). Voxelotor concentration also had a significant effect on increases in hemoglobin (R2 = 0.40, P < .001). CONCLUSIONS: Thus, peak splitting CZE and HPLC can be used to estimate voxelotor concentration.

Filaggrin Gene Mutation in Pediatric Patients with Atopic Dermatitis: A Look into Indian Gene Pool, a Pilot Study.

Background: Mutations in the filaggrin (FLG) gene has been reported to be an indicator of poor prognosis of atopic dermatitis (AD). It has been reported that there is a considerable variation in the mutations detected in the FLG gene in different ethnicities. Aim: To detect the presence of mutations in the FLG gene in pediatric subjects with atopic dermatitis (AD) and to compare the detected mutations with those already reported from different ethnicities. Materials and Methods: Genomic DNA extracted using standard procedure from peripheral venous blood of 30 patient and 15 control samples. Sequence analysis of the FLG gene carried out and detected changes was then cross referenced with those mutations already reported to check for novelty of detected changes. Results: Amino acid changes were detected in 28 of the patient samples and in none of the control samples indicating that changes in the FLG gene were more common in the patient group than the control group (Fishers exact test, P < 0.0001). The most commonly reported mutations R501X and 2282del4 were not detected. Only 5 of the detected 22 amino acid changes H2507Q, L2481S, K2444E, E2398Q, and S2366T have been previously reported and are not clinically significant; however, in one patient a stop codon was detected (S2366STOP). P2238N, R2239W, and V2243L detected in 70% of the samples and S2231E detected in 67% of the patient samples have not been reported so far and their clinical significance is yet to be analyzed. Conclusion: Analyses of mutations already reported showed that the changes detected from this study are novel to Indian traits. While this adds on to the minimal data available from the Indian subcontinent further analyses has to be carried out to analyze the pathogenicity of these detected changes on larger samples sizes. Aim: To detect the presence of mutations in the FLG gene in pediatric subjects with atopic dermatitis (AD) and to compare the detected mutations with those already reported from different ethnicities.

A framework for perioperative care for lower extremity vascular bypasses: A Consensus Statement by the Enhanced Recovery after Surgery (ERAS®) Society and Society for Vascular Surgery.

The Society for Vascular Surgery and the Enhanced Recovery After Surgery (ERAS) Society formally collaborated and elected an international, multi-disciplinary panel of experts to review the literature and provide evidence-based suggestions for coordinated perioperative care for patients undergoing infrainguinal bypass surgery for peripheral artery disease. Structured around the ERAS core elements, 26 suggestions were made and organized into preadmission, preoperative, intraoperative, and postoperative sections.

Nanovesicle and extracellular polymeric substance synthesis from the remediation of heavy metal ions from soil.

Heavy metal toxicity affects aquatic plants and animals, disturbing biodiversity and ecological balance causing bioaccumulation of heavy metals. Industrialization and urbanization are inevitable in modern-day life, and control and detoxification methods need to be accorded to meet the hazardous environment. Microorganisms and plants have been widely used in the bioremediation of heavy metals. Sporosarcina pasteurii, a gram-positive bacterium that is widely known for its calcite precipitation property in bio-cementing applications has been explored in the study for its metal tolerance ability for the first time. S. pasteurii SRMNP1 (KF214757) can tolerate silver stress to form nanoparticles and can remediate multiple heavy metals to promote the growth of various plants. This astounding property of the isolate warranted extensive examinations to comprehend the physiological changes during an external heavy metal stress condition. The present study aimed to understand various physiological responses occurring in S. pasteuriiSRMNP1 during the metal tolerance phenomenon using electron microscopy. The isolate was subjected to heavy metal stress, and a transmission electron microscope examination was used to analyze the physiological changes in bacteria to evade the metal stress. S. pasteurii SRMNP1 was tolerant against a wide range of heavy metal ions and can withstand a broad pH range (5-9). Transmission Electron Microscopy (TEM) examination of S. pasteurii SRMNP1 followed by 5 mM nickel sulfate treatment revealed the presence of nanovesicles encapsulating nanosized particles in intra and extracellular spaces. This suggests that the bacteria evade the metal stress by converting the metal ions into nanosized particles and encapsulating them within nanovesicles to efflux them through the vesicle budding mechanism. Moreover, the TEM images revealed an excessive secretion of extracellular polymeric substances by the strain to discharge the metal particles outside the bacterial system. S. pasteurii can be foreseen as an effective bioremediation agent with the potential to produce nanosized particles, nanovesicles, and extracellular polymeric substances. This study provides physiological evidence that, besides calcium precipitation applications, S. pasteurii can further be explored for its multidimensional roles in the fields of drug delivery and environmental engineering.

Excess deaths among adults with sickle cell disease in 2020 compared to prior years.

In 2020, there was a 20% increase in excess deaths in the USA due to COVID infections but also to changes in the healthcare system due to the pandemic. We hypothesized that people living with sickle cell disease (SCD) may be vulnerable to these changes as SCD can lead to rapid decompensation. We examined all deaths of people with SCD at our center in 2020. Cause of death was determined, clinical variables, and healthcare utilization, and the presence of COVID infection, sepsis, and acute organ failure during the death event was obtained from the electronic medical record. Deaths in 2020 were compared to deaths in 2017-2019. In 2020, deaths increase 244% (22 vs 9), but acute or previous COVID infections were identified in only 36% of 2020 deaths. People who died in 2020 were more likely to have developed acute organ failure during the death event (70.6% vs 21.1%, p = 0.003) compared to prior years. They were also more likely to have a history of stroke and more frequent hematology clinic visits. Deaths in 2020 doubled compared to prior years and COVID infection could not account for all of this excess mortality. People who died in 2020 may have had more severe disease as suggested by having more clinic visits and higher rates of stroke and were more likely to develop organ failure during the death event. This demonstrates that people with SCD may be especially vulnerable to delays in care. Larger multicenter studies should be conducted to examine this further.

Dissemination of Cardiopulmonary Resuscitation Training for Nurses Treating Coronavirus Disease-2019 Patients: A Single-arm Pre-experimental Study.

Introduction: Providing cardiopulmonary resuscitation (CPR) to a coronavirus disease-2019 (COVID-19) patient is challenging for the healthcare providers (HCP). COVID-19 cases have specific CPR requirements, which differ from standard resuscitation efforts. Objective: This paper aims to evaluate whether online learning can be an effective methodology for imparting information on the management of cardiopulmonary arrest in COVID-19 patients due to a novel virus unknown to HCP based on evaluation of knowledge acquisition and satisfaction of the nurses in the new area of medicine and virology. Methodology: In a single-arm, one group before and after design, from a cohort of 160 nurses trained in comprehensive cardiac life support (CCLS) formulated by the Indian Resuscitation Council (IRC), 73 nurses participated in the study. After obtaining informed consent through the email from the nurses, baseline data including demographic profile and knowledge related to CPR in COVID-19 patients were collected. An online intervention spread over 1 week was given using a validated e-learning module. The online intervention was found to be effective (pre- and post-intervention knowledge score 13.65 ± 3.01 vs 19.92 ± 1.94, p = 0.001). The majority of nurses were highly satisfied with the content and the training methodology (37.23 ± 4.70). Conclusion: A well-structured, online study material can be used in imparting knowledge and demonstrating the basic and essential skills to nurses, required for giving CPR to COVID-19 patients. How to cite this article: Joshi P, Das S, Thomas M, Mawar S, Garg R, Shariff A, et al. Dissemination of Cardiopulmonary Resuscitation Training for Nurses Treating Coronavirus Disease-2019 Patients: A Single-arm Pre-experimental Study. Indian J Crit Care Med 2022;26(3):327-330.

Perioperative care in open aortic vascular surgery: A consensus statement by the Enhanced Recovery After Surgery (ERAS) Society and Society for Vascular Surgery.

The Society for Vascular Surgery and the Enhanced Recovery After Surgery Society formally collaborated and elected an international, multidisciplinary panel of experts to review the literature and provide evidence-based recommendations related to all the health care received in the perioperative period for patients undergoing open abdominal aortic operations (both transabdominal and retroperitoneal approaches, including supraceliac, suprarenal, and infrarenal clamp sites) for aortic aneurysm and aortoiliac occlusive disease. Structured around the Enhanced Recovery After Surgery core elements, 36 recommendations were made and organized into preadmission, preoperative, intraoperative, and postoperative recommendations.

Mothers' Knowledge of Childhood Neurodevelopmental Disorders: Indian Perspective.

Objectives This study aims to assess the knowledge of childhood neurodevelopmental disorders (CNDDs) among mothers of children younger than 5 years and to promote awareness through health education to promote early identification. Methods Quantitative approach, a descriptive survey in nature research design, was used. A total of 173 mothers who contented the inclusion conditions were chosen as sample on the basis of purposive sampling. The research study was done at the selected Primary Health Centre, Bengaluru, Karnataka. The tabulations were measured and construed based on the objectives of the study by using descriptive and inferential statistics. Results The findings showed that most of the mothers have insufficient knowledge (mean and standard deviation: 3.02 ± 2.75) and there is no substantial relationship found between mothers' knowledge on CNDD and their demographic variables except their occupation and majority of the mothers communicated that they needed further facts toward CNDD. Conclusion The government and nongovernmental organizations can take initiations to conduct health education programs toward childhood developmental delays and disorders for the general public including mothers and community health workers.

Electrospun polylactic acid-chitosan composite: a bio-based alternative for inorganic composites for advanced application.

Fabricating novel materials for biomedical applications mostly require the use of biodegradable materials. In this work biodegradable materials like polylactic acid (PLA) and chitosan (CHS) were used for designing electrospun mats. This work reports the physical and chemical characterization of the PLA-CHS composite, prepared by the electrospinning technique using a mixed solvent system. The addition of chitosan into PLA, offered decrease in fiber diameter in the composites with uniformity in the distribution of fibers with an optimum at 0.4wt% CHS. The fiber formation and the reduction in fiber diameter were confirmed by the SEM micrograph. The inverse gas chromatography and contact angle measurements supported the increase of hydrophobicity of the composite membrane with increase of filler concentration. The weak interaction between PLA and chitosan was confirmed by Fourier transform infrared spectroscopy and thermal analysis. The stability of the composite was established by zeta potential measurements. Cytotoxicity studies of the membranes were also carried out and found that up to 0.6% CHS the composite material was noncytotoxic. The current findings are very important for the design and development of new materials based on polylactic acid-chitosan composites for environmental and biomedical applications.

Computational analysis of ribonomics datasets identifies long non-coding RNA targets of γ-herpesviral miRNAs.

Ribonomics experiments involving crosslinking and immuno-precipitation (CLIP) of Ago proteins have expanded the understanding of the miRNA targetome of several organisms. These techniques, collectively referred to as CLIP-seq, have been applied to identifying the mRNA targets of miRNAs expressed by Kaposi's Sarcoma-associated herpes virus (KSHV) and Epstein-Barr virus (EBV). However, these studies focused on identifying only those RNA targets of KSHV and EBV miRNAs that are known to encode proteins. Recent studies have demonstrated that long non-coding RNAs (lncRNAs) are also targeted by miRNAs. In this study, we performed a systematic re-analysis of published datasets from KSHV- and EBV-driven cancers. We used CLIP-seq data from lymphoma cells or EBV-transformed B cells, and a crosslinking, ligation and sequencing of hybrids dataset from KSHV-infected endothelial cells, to identify novel lncRNA targets of viral miRNAs. Here, we catalog the lncRNA targetome of KSHV and EBV miRNAs, and provide a detailed in silico analysis of lncRNA-miRNA binding interactions. Viral miRNAs target several hundred lncRNAs, including a subset previously shown to be aberrantly expressed in human malignancies. In addition, we identified thousands of lncRNAs to be putative targets of human miRNAs, suggesting that miRNA-lncRNA interactions broadly contribute to the regulation of gene expression.

Modified Cross-Linking, Ligation, and Sequencing of Hybrids (qCLASH) Identifies Kaposi's Sarcoma-Associated Herpesvirus MicroRNA Targets in Endothelial Cells.

Kaposi's sarcoma (KS) tumors are derived from endothelial cells and express Kaposi's sarcoma-associated herpesvirus (KSHV) microRNAs (miRNAs). Although miRNA targets have been identified in B cell lymphoma-derived cells and epithelial cells, little has been done to characterize the KSHV miRNA targetome in endothelial cells. A recent innovation in the identification of miRNA targetomes, cross-linking, ligation, and sequencing of hybrids (CLASH), unambiguously identifies miRNAs and their targets by ligating the two species while both species are still bound within the RNA-induced silencing complex (RISC). We developed a streamlined quick CLASH (qCLASH) protocol that requires a lower cell input than the original method and therefore has the potential to be used on patient biopsy samples. Additionally, we developed a fast-growing, KSHV-negative endothelial cell line derived from telomerase-immortalized vein endothelial long-term culture (TIVE-LTC) cells. qCLASH was performed on uninfected cells and cells infected with either wild-type KSHV or a mutant virus lacking miR-K12-11/11*. More than 1,400 cellular targets of KSHV miRNAs were identified. Many of the targets identified by qCLASH lacked a canonical seed sequence match. Additionally, most target regions in mRNAs originated from the coding DNA sequence (CDS) rather than the 3' untranslated region (UTR). This set of genes includes some that were previously identified in B cells and some new genes that warrant further study. Pathway analysis of endothelial cell targets showed enrichment in cell cycle control, apoptosis, and glycolysis pathways, among others. Characterization of these new targets and the functional consequences of their repression will be important in furthering our understanding of the role of KSHV miRNAs in oncogenesis.IMPORTANCE KS lesions consist of endothelial cells latently infected with KSHV. Cells that make up these lesions express KSHV miRNAs. Identification of the targets of KSHV miRNAs will help us understand their role in viral oncogenesis. The cross-linking and sequencing of hybrids (CLASH) protocol is a method for unambiguously identifying miRNA targetomes. We developed a streamlined version of CLASH, called quick CLASH (qCLASH). qCLASH requires a lower initial input of cells than for its parent protocol. Additionally, a new fast-growing KSHV-negative endothelial cell line, named TIVE-EX-LTC cells, was established. qCLASH was performed on TIVE-EX-LTC cells latently infected with wild-type (WT) KSHV or a mutant virus lacking miR-K12-11/11*. A number of novel targets of KSHV miRNAs were identified, including targets of miR-K12-11, the ortholog of the cellular oncogenic miRNA (oncomiR) miR-155. Many of the miRNA targets were involved in processes related to oncogenesis, such as glycolysis, apoptosis, and cell cycle control.

Staff Nurses Eliminated Catheter-Associated Urinary Tract Infections in a Neurosurgical Intensive Care Unit.

Catheter-associated urinary tract infections (CAUTIs) are among the leading causes of health care-associated infections in neurosurgical populations. Successful reduction of CAUTIs involve the development of staff nurses as front-line change agents equipped with preventative strategies, educational interventions, and sustainable maintenance for positive patient outcomes.

Four-headed biceps and triceps brachii muscles, with neurovascular variation.

Anatomical variations of the biceps brachii and triceps brachii have been described by various authors, but the occurrence of four-headed biceps brachii and triceps brachii in an ipsilateral arm is rare and has not been reported before in the literature. During routine cadaveric dissection in the department of anatomy, Kasturba Medical College, Mangalore, India, various unusual variants were noted in the left arm of a cadaver of a 67-year-old man. The variants include a four-headed biceps, a four-headed triceps, communication between the musculocutaneous and median nerves, and a high origin of deep brachial artery from the axillary artery. The occurrence of these anomalies and the clinical and morphological significance are discussed.

Multiple variations of the extensor tendons of the forearm.

Anatomic variations of the extensor tendons of the forearm are frequent and its knowledge is important to assess the diseased and traumatized hand. During routine cadaveric dissection in the Department of Anatomy, Kasturba Medical College, Mangalore, India, we came across unusual variations in the left upper limb of a 51-years-old male cadaver. The variants are, the complete absence of the extensor pollicis brevis (EPB), the abductor pollicis longus (APL) having six slips of insertion with additional muscle slip from the brachio-radialis (BR). The extensor digitorum communis (EDC) had five tendon slips, the extra tendon splits close to the metacarpo-phalangeal (MP) joint and contribute to the tendons of the ring and little fingers. The number of accessory tendons of APL has functional significance in the development of de Quervain's stenosing tendovaginitis. The present case is unique in the sense that, all the three variations are present in the ipsilateral upper limb. The occurrence of these anomalies and its clinical and embryological significance are discussed.