A synthetic bioactive peptide of the C-terminal fragment of adhesion protein Fibulin7 attenuates the inflammatory functions of innate immune cells in LPS-induced systemic inflammation.

OBJECTIVE: Systemic inflammation is associated with improper localization of hyperactive neutrophils and monocytes in visceral organs. Previously, a C-terminal fragment of adhesion protein Fibulin7 (Fbln7-C) was shown to regulate innate immune functionality during inflammation. Recently, a shorter bioactive peptide of Fbln7-C, FC-10, via integrin binding was shown to reduce ocular angiogenesis. However, the role of FC-10 in regulating the neutrophils and monocyte functionality during systemic inflammatory conditions is unknown. The study sought to explore the role of FC-10 peptide on the functionality of innate immune cells during inflammation and endotoxemic mice. METHODS: Neutrophils and monocytes were isolated from healthy donors and septic patient clinical samples and Cell adhesion assay was performed using a UV spectrophotometer. Gene expression studies were performed using qPCR. Protein level expression was measured using ELISA and flow cytometry. ROS assay, and activation markers analysis in vitro, and in vivo were done using flow cytometry. TREATMENT: Cells were stimulated with LPS (100 ng/mL) and studied in the presence of peptides (10 µg, and 20 µg/mL) in vitro. In an in vivo study, mice were administered with LPS (36.8 mg/kg bw) and peptide (20 µg). RESULTS: This study demonstrates that human neutrophils and monocytes adhere to FC-10 via integrin ß1, inhibit spreading, ROS, surface activation markers (CD44, CD69), phosphorylated Src kinase, pro-inflammatory genes, and protein expression, compared to scrambled peptide in cells isolated from healthy donors and clinical sample. In line with the in vitro data, FC-10 (20 µg) administration significantly decreases innate cell infiltration at inflammatory sites, improves survival in endotoxemia animals & reduces the inflammatory properties of neutrophils and monocytes isolated from septic patients. CONCLUSION: FC-10 peptide can regulate neutrophils and monocyte functions and has potential to be used as an immunomodulatory therapeutic in inflammatory diseases.

Role of RhoG as a regulator of cellular functions: integrating insights on immune cell activation, migration, and functions.

BACKGROUND: RhoG is a multifaceted member of the Rho family of small GTPases, sharing the highest sequence identity with the Rac subfamily members. It acts as a molecular switch, when activated, plays a central role in regulating the fundamental processes in immune cells, such as actin-cytoskeleton dynamics, transendothelial migration, survival, and proliferation, including immunological functions (e.g., phagocytosis and trogocytosis) during inflammatory responses. METHOD: We have performed a literature review based on published original and review articles encompassing the significant effect of RhoG on immune cell functions from central databases, including PubMed and Google Scholar. RESULTS AND CONCLUSIONS: Recently published data shows that the dynamic expression of different transcription factors, non-coding RNAs, and the spatiotemporal coordination of different GEFs with their downstream effector molecules regulates the cascade of Rho signaling in immune cells. Additionally, alterations in RhoG-specific signaling can lead to physiological, pathological, and developmental adversities. Several mutations and RhoG-modulating factors are also known to pre-dispose the downstream signaling with abnormal gene expression linked to multiple diseases. This review focuses on the cellular functions of RhoG, interconnecting different signaling pathways, and speculates the importance of this small GTPase as a prospective target against several pathological conditions.

Modified Bain's Circuit as an Alternate to Non-invasive Ventilation in COVID 19.

Introduction COVID-19 is a pandemic that severely affects the lungs. Symptomatically affected individuals often become severely hypoxic, requiring non-invasive ventilation. The scarcity of resources in resource-compromised countries like India led to the adoption of novel strategies like using Bain's circuit for assisting spontaneous ventilation. This study compares the outcome when a standard circuit is replaced with a shortened Bain's circuit. Aims and objectives To compare shortened Bain's circuit and bilevel positive airway pressure (BiPAP) in spontaneously ventilated COVID 19 patients with regards to effects on hemodynamic stability and efficacy of ventilation using blood gas analysis. Methodology Twenty-four COVID patients aged between 35-70 years, requiring non-invasive ventilation but not tolerating BiPAP or not improving on BiPAP were enrolled in the study. Baseline heart rate and arterial blood gases (ABG) were recorded. Patients were then ventilated using shortened Bain's circuit. Heart rate and ABG were then recorded two hours after ventilation. Results Hemodynamic and blood gas parameters were comparable between the two groups at baseline and on BiPAP. Group A showed better hemodynamic and blood gas profiles compared to group B, but the difference was not statistically significant because of small sample size. Conclusion Shortened Bain's circuit may be a viable alternative to non-invasive ventilation in spontaneously breathing hypoxic patients with efficacy comparable to a standard Bain's circuit and reduced chances of carbon dioxide retention. Studies with a larger sample size are needed to further validate the conclusion.

Comparative Study Among Ketamine, Fentanyl, and Ropivacaine, as Pre-incisional Analgesic Given by Surgical Site Infiltration, in Cases Posted for Elective Lower Segment Cesarean Section Under General Anesthesia.

Aim The aim of this study was to evaluate the effect of surgical site infiltration prior to giving incision, with ketamine, fentanyl, and ropivacaine, on perioperative hemodynamic parameters, pain, and any adverse outcomes. Methods The study was carried out on 68 patients divided into four groups of 17 each. After intubation, the study drug was infiltrated subcutaneously at the surgical site, seven min before incision, by the surgeon who was unaware of the injectate. Hemodynamic variables were monitored and recorded during the intraoperative period and postoperative period at one, two, four, six, eight, 12, and 24 hours. The level of sedation was assessed using the Ramsay sedation score (RSS). The fetal outcome was assessed by the APGAR (Appearance, Pulse, Grimace, Activity, and Respiration) score at one, five, and 10 minutes. Results The mean visual analog scale (VAS) scores were significantly (p < 0.05) lower in group A as compared to other groups for most of the postoperative period. Hemodynamic stability was also significantly better with group A as compared to other groups during the perioperative period. The fetal outcome was comparable between the study groups with the APGAR scores similar between the groups at 10 min. However, significantly lower APGAR values were seen in group C as compared to the other study groups, at one and five minutes. Conclusion Ketamine infiltration resulted in a better hemodynamic profile, a significantly longer duration of the postoperative pain-free interval, and more sedation in the postoperative anesthesia care unit (PACU). It did not affect the fetal wellbeing, as discernable from the APGAR scores.

Viral Ribonucleic Acid Shedding and Transmission Potential of Asymptomatic and Paucisymptomatic Coronavirus Disease 2019 Patients.

We studied the pattern and duration of viral ribonucleic acid (RNA) shedding in 32 asymptomatic and 11 paucisymptomatic coronavirus disease 2019 cases. Viral RNA shedding in exhaled breath progressively diminished and became negative after 6 days of a positive reverse-transcription polymerase chain reaction test. Therefore, the duration of isolation can be minimized to 6 days.

Transcriptome Analysis Reveals Important Candidate Genes Related to Nutrient Reservoir, Carbohydrate Metabolism, and Defence Proteins during Grain Development of Hexaploid Bread Wheat and Its Diploid Progenitors.

Wheat grain development after anthesis is an important biological process, in which major components of seeds are synthesised, and these components are further required for germination and seed vigour. We have made a comparative RNA-Seq analysis between hexaploid wheat and its individual diploid progenitors to know the major differentially expressed genes (DEGs) involved during grain development. Two libraries from each species were generated with an average of 55.63, 55.23, 68.13, and 103.81 million reads, resulting in 79.3K, 113.7K, 90.6K, and 121.3K numbers of transcripts in AA, BB, DD, and AABBDD genome species respectively. Number of expressed genes in hexaploid wheat was not proportional to its genome size, but marginally higher than that of its diploid progenitors. However, to capture all the transcripts in hexaploid wheat, sufficiently higher number of reads was required. Functional analysis of DEGs, in all the three comparisons, showed their predominance in three major classes of genes during grain development, i.e., nutrient reservoirs, carbohydrate metabolism, and defence proteins; some of them were subsequently validated through real time quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR). Further, developmental stage-specific gene expression showed most of the defence protein genes expressed during initial developmental stages in hexaploid contrary to the diploids at later stages. Genes related to carbohydrates anabolism expressed during early stages, whereas catabolism genes expressed at later stages in all the species. However, no trend was observed in case of different nutrient reservoirs gene expression. This data could be used to study the comparative gene expression among the three diploid species and homeologue-specific expression in hexaploid.

SATB1-regulated transcriptome datasets of Rcho-1 rat trophoblast stem cells.

SATB homeobox 1 (SATB1) and its heterodimeric partner SATB2 play an important regulatory role in maintaining proliferation of trophoblast stem (TS) cells and in inhibiting trophoblast differentiation. To identify the SATB-regulated genes in TS cells, we studied the transcriptome changes in a 'loss of function' model of Rcho-1 rat TS cell line. Satb1 gene expression was silenced by lentiviral delivery of shRNAs targeted to exon 9 and exon 12. An Egfp shRNA was used as a non-targeted control. Total RNA was purified from shRNA-transduced Rcho-1 cells, and whole transcriptome was assessed by RNA-sequencing on an Illumina HiSeq X platform. Differentially expressed genes in Satb1 shRNA-transduced cells were identified by analyses of the RNA-sequencing data using CLC Genomics Workbench. Differentially expressed genes with each of the two different shRNAs were compared to identify SATB1-target genes and to eliminate the potential off-targets of the shRNAs. These datasets can be used to identify the SATB-regulated genes in TS cells and to understand the molecular mechanisms that regulate trophoblast proliferation and inhibit differentiation.

Transcriptome data of cultivated tetraploid and hexaploid wheat variety during grain development.

Wheat is a major food crop and an important component of human diet throughout the world. There are two major types of cultivated wheat; one is tetraploid durum (pasta) wheat and another one is hexaploid bread wheat. Wheat grain is the reservoir of two major dietary components - carbohydrate and protein, which get accumulated during seed maturation and directly affects yield and quality. Hexaploid, having 6 copies of each chromosome differs to a great extent from tetraploid having 4 copies of each chromosome. Studying the gene expression pattern in developing grain would help in understanding the difference in metabolic process as well as involvement of the genes in these two types of wheat. A transcriptional comparison of developing grains was carried out between the two wheat genotypes; tetraploid (AABB:PDW233) and hexaploid (AABBDD:PBW343) using RNA-seq. Approximately 194 million raw reads were obtained from both libraries. After removal of contaminations, a huge proportion (>99%), of high quality reads were obtained, were aligned to reference genome. A total of 2324 up-regulated and 522 down-regulated genes were identified as differentially expressed between PDW233 vs PBW343. Gene ontology annotation and enrichment analysis gave further information about differentially expressed genes between durum and bread wheat. This information will help in understanding process grain reserve in tetraploid and hexaploid wheat in relation to their nutritional quality.