Systems genetics identifies miRNA-mediated regulation of host response in COVID-19.

BACKGROUND: Individuals infected with SARS-CoV-2 vary greatly in their disease severity, ranging from asymptomatic infection to severe disease. The regulation of gene expression is an important mechanism in the host immune response and can modulate the outcome of the disease. miRNAs play important roles in post-transcriptional regulation with consequences on downstream molecular and cellular host immune response processes. The nature and magnitude of miRNA perturbations associated with blood phenotypes and intensive care unit (ICU) admission in COVID-19 are poorly understood. RESULTS: We combined multi-omics profiling-genotyping, miRNA and RNA expression, measured at the time of hospital admission soon after the onset of COVID-19 symptoms-with phenotypes from electronic health records to understand how miRNA expression contributes to variation in disease severity in a diverse cohort of 259 unvaccinated patients in Abu Dhabi, United Arab Emirates. We analyzed 62 clinical variables and expression levels of 632 miRNAs measured at admission and identified 97 miRNAs associated with 8 blood phenotypes significantly associated with later ICU admission. Integrative miRNA-mRNA cross-correlation analysis identified multiple miRNA-mRNA-blood endophenotype associations and revealed the effect of miR-143-3p on neutrophil count mediated by the expression of its target gene BCL2. We report 168 significant cis-miRNA expression quantitative trait loci, 57 of which implicate miRNAs associated with either ICU admission or a blood endophenotype. CONCLUSIONS: This systems genetics study has given rise to a genomic picture of the architecture of whole blood miRNAs in unvaccinated COVID-19 patients and pinpoints post-transcriptional regulation as a potential mechanism that impacts blood traits underlying COVID-19 severity. The results also highlight the impact of host genetic regulatory control of miRNA expression in early stages of COVID-19 disease.

A Comparative Study of Craniosonogram and CT scan of Brain in CNS Complication of Perinatal Asphyxia.

Pereinatal asphyxia is one of the most important complications related with the process of birth and this complications affect not only the brain but also many other organs. The purpose of this cross sectional study is to compare the role of craniosonogram and CT scan of the brain to delineate the cerebral pathology in respondent of preterm and term infant and to assess the Kappa test for agreement. This study was carried out in the department of Radiology & Imaging of Mymensingh Medical College Hospital, during the period of July 2015 to June 2017. A total number of 40 neonates clinically diagnosed as perinatal asphyxia referred for Craniosonogram and Computed tomography (CT) were included in this study. The test of agreement of USG in detection of neonatal cerebral pathology was calculated. Male to female ratio was 1:1.4. More than half (55.0%) patients were preterm age. The mean birth weight was found 2.3±0.6 kg and mean age was 22.7±12.7 days. Fifty five percent respondent mothers had antenatal check up and 27% had anemia, 11% had premature rupture of membrane, 9% had multiple pregnancy and 7% patient had hypertension. Cerebral pathology was found 28 and 31 cases by Craniosonogram and CT scan respectively. Germinal matrix hemorrhage/IVH (Intra ventricular hemorrhage) found 7(17.5%) in USG and 4(10.0%) in CT scan. Hypoxic ischaemic changes with mild ventriculomegaly observed 5(12.5%) in USG and 7(17.5%) in CT scan. Hypoxic ischaemic change found 4(10.0%) in USG and 5(12.5%) in CT scan. In USG evaluation of 28 patients having cerebral pathology & 16(72.7%) had in preterm group and 12(66.7%) in term group. In CT scan of brain 31 patients with cerebral pathology & 15(37.5%) in preterm group and 16(40.0%) in term group. CT scan found cerebral pathology 77.5% (31/40) cases and USG found 70.0% (28/40), with Kappa value was 0.551, which indicates that fair agreement between USG and CT scan for detection of cerebral pathology in respondent. Craniosonogram is a useful method in all neonates specially preterm to see the CNS complication of perinatal asphyxia.

The multidrug resistance pump ABCB1 is a substrate for the ubiquitin ligase NEDD4-1.

The ATP Binding Cassette transporter ABCB1 can export the neurotoxic peptide ß-amyloid from endothelial cells that line the blood-brain barrier (BBB). This has the potential to lower cerebral levels of ß-amyloid, but ABCB1 expression in the BBB appears to be progressively reduced in patients with Alzheimer's disease. The surface density of many membrane proteins is regulated by ubiquitination catalyzed by ubiquitin E3 ligases. In brain capillaries of mice challenged with ß-amyloid ex vivo, we show that the level of the ubiquitin ligase Nedd4 increases concomitant with reduction in Abcb1. In vitro we show that human ABCB1 is a substrate for human NEDD4-1 ligase. Recombinant ABCB1 was purified from Sf21 insect cells and incubated with recombinant NEDD4-1 purified from Escherichia coli. The treated ABCB1 had reduced mobility on SDS-PAGE, and mass spectrometry identified eight lysine residues, K271, K272, K575, K685, K877, K885, K887 and K1062 that were ubiquitinated by NEDD4-1. Molecular modelling showed that all of the residues are exposed on the surface of the intracellular domains of ABCB1. K877, K885 and K887 in particular, are located in the intracellular loop of transmembrane helix 10 (TMH10) in close proximity, in the tertiary fold, to a putative NEDD4-1 binding site in the intracellular helix extending from TMH12 (PxY motif, residues 996-998). Transient expression of NEDD4-1 in HEK293 Flp-In cells stably expressing ABCB1 was shown to reduce the surface density of the transporter. Together, the data identify this ubiquitin ligase as a potential target for intervention in the pathophysiology of Alzheimer's disease.

The Q loops of the human multidrug resistance transporter ABCB1 are necessary to couple drug binding to the ATP catalytic cycle.

For a primary active pump, such as the human ATP-binding-cassette (ABC) transporter ABCB1, coupling of drug-binding by the two transmembrane domains (TMDs) to the ATP catalytic cycle of the two nucleotide-binding domains (NBDs) is fundamental to the transport mechanism, but is poorly understood at the biochemical level. Structure data suggest that signals are transduced through intracellular loops of the TMDs that slot into grooves on the NBDs. At the base of these grooves is the Q loop. We therefore mutated the eponymous glutamine in one or both NBD Q loops and measured the effect on conformation and function by using a conformation-sensitive antibody (UIC2) and a fluorescent drug (Bodipy-verapamil), respectively. We showed that the double mutant is trapped in the inward-open state, which binds the drug, but cannot couple to the ATPase cycle. Our data also describe marked redundancy within the transport mechanism, because single-Q-loop mutants are functional for Bodipy-verapamil transport. This result allowed us to elucidate transduction pathways from twin drug-binding cavities to the Q loops using point mutations to favor one cavity over the other. Together, the data show that the Q loop is the central flexion point where the aspect of the drug-binding cavities is coupled to the ATP catalytic cycle.

(3R*,4R*,5S*)-4-(4-Methyl-phen-yl)-2,3-diphenyl-7-[(R*)-1-phenyl-ethyl]-1-oxa-2,7-diaza-spiro-[4.5]decan-10-one oxime.

In the title compound, C(34)H(35)N(3)O(2), the polysubstituted piperidine ring adopts a chair conformation and the isoxazolidine ring is in an envelope form. The mol-ecules are linked into a chain along the b axis by O-H⋯N, C-H⋯O and C-H⋯N inter-actions. The chains are cross-linked via weak C-H⋯π inter-actions.

Increased acute GvHD and higher transplant-related mortality in non-caucasians undergoing standard sibling allogeneic stem cell transplantation.

We conducted a retrospective study to compare outcome in Caucasians and non-Caucasians undergoing standard sibling allogeneic SCT. End points of the study were to compare graft-versus-host disease (GvHD) occurrence and transplant-related mortality (TRM). There were 251 patients, 43 non-Caucasian and 208 Caucasian. A higher proportion of non-Caucasian patients developed acute GvHD (aGvHD) grade 2 or greater as compared to the Caucasian group (48 vs 26%, respectively) P = 0.02. With a median follow-up of 27 months, 26% (11/43) of non-Caucasians and 14% (29/208) of Caucasian patients had died from TRM, which accounted for 55% of all deaths in the non-Caucasian group compared to 33% in Caucasians, P = 0.02. Overall survival 12 months post transplant was 64 vs 69% in the non-Caucasian and Caucasian groups, respectively (P = 0.43). Although there were higher numbers of CMV-positive patients in the non-Caucasian group, there were no deaths from CMV reactivation in this subgroup. We conclude that there is increased TRM and aGvHD following standard sibling allograft in the non-Caucasian population and this could be due to either differences in tumour biology or extrinsic factors such as socio-economic factors, nutritional status, post transplant care or presenting with late stage disease.

Stem cell transplantation after salvage therapy with high-dose cytarabine and amsacrine in adults with high-risk leukaemia.

Stem cell transplantation (SCT) may be the only curative option for patients with relapsed or refractory leukaemia, that is, high-risk (HR) leukaemia. Several salvage regimens have been used to cytoreduce disease before SCT, but disease progression or treatment toxicity limits numbers of patients receiving SCT. Here, we report our experience with high-dose cytarabine and amsacrine (Ara-amsa) to salvage patients with HR-leukaemia in the context of SCT. A total of 34 patients with HR-leukaemia (20 AML, 12 ALL, two advanced CML) received 3 g/m(2)/day cytarabine for 5 days and amsacrine 200 mg/m(2)/day for 3 days. Disease response was observed in 62% of patients. Toxicity was limited to neutropenic fever, one patient developed cerebellar toxicity and there was one treatment-related death. A total of 17 patients proceeded to SCT (12 allografts and five autografts). Median survival (OS) of all patients was 10.8 months (95% CI 7.8-21). Patients who were consolidated with SCT after salvage therapy had a superior median OS of 29.4 months (95% CI 12.5-upper limit not reached, n=17) than those who did not receive SCT (6.7 months, CI 1.5-8.6, P<0.0001). Median disease-free survival with SCT (23 months) was higher than after treatment with salvage chemotherapy alone (6.7 months, P=0.0002). Thus Ara-amsa can be used effectively to salvage HR-leukaemia, enabling further consolidation with SCT.

Development and validation of a questionnaire for the assessment of bowel and lower urinary tract symptoms in women.

OBJECTIVE: To develop a simple but sensitive instrument to evaluate and document symptoms of both bowel and urinary dysfunction in women. DESIGN: A 22-item questionnaire covering a range of bowel and urinary symptoms was developed and underwent rigorous psychometric testing. SETTING: The gynaecology departments of three hospitals, a urogynaecology clinic, a functional bowel clinic and a general practice. POPULATION: Six hundred and thirty women, comprising four groups: 1. women awaiting hysterectomy (n = 379), 2. women following hysterectomy (n = 45), 3. women referred with functional bowel and/or urinary symptoms (n = 65), 4. asymptomatic controls (n = 141). MAIN OUTCOME MEASURES: The content, construct and criterion validity, internal consistency, reliability and responsiveness of the questionnaire were measured. RESULTS Peer and patient reports and missing data patterns supported face and content validity. Factor analysis showed a clinically relevant four-factor structure with low content replication able to distinguish between patient groups, indicating good internal structure. Comparison with clinical, anorectal physiological, videoproctographic, transit time and urodynamic test results provide provisional indication of criterion validity. Key domain question analysis and Cronbach's alphas showed internal consistency. Kappa values demonstrated good test-retest reliability and key question correlation over time proved responsiveness. CONCLUSIONS: Our findings support the suitability, clinical validity, reliability and responsiveness of a simple questionnaire, which is sensitive to the constraints of clinical practice. The authors recommend its use in health care evaluation research assessing the effects of pelvic surgery and as a useful tool in comparing treatment efficacy.

Delay and survival in bladder cancer.

OBJECTIVE: To assess in detail and evaluate the effect on survival of delays in the diagnosis and treatment of cancer (which might lead to a worse prognosis), dividing the delay from onset of symptoms to first treatment into several components, comprising patient delay, general practitioner (GP) delay, and two or more periods of hospital delay. PATIENTS AND METHODS: Data were prospectively collected on 1537 new cases of urothelial cancer in the West Midlands from 1 January 1991 to 30 June 1992. Death information was obtained from the West Midlands Cancer Intelligence Unit and censored at 31 July 2000. The influence of delay times on survival was explored. RESULTS: The median delay from onset of symptoms to GP referral was 14 days (Delay 1), from GP referral to first hospital attendance was 28 days (Delay 2), and from first hospital attendance to first transurethral resection of bladder tumour was 20 days (Delay 3). The median hospital delay (Delay 2 + 3) was 68 days and the median total delay (Delay 1 + 2 + 3) was 110 days. Patients with a shorter Delay 1 had a lower tumour stage and a 5% better 5-year survival. Patients with a shorter hospital delay had worse survival; total delay had no effect on survival. CONCLUSIONS: There was significantly better survival for patients referred to hospital within 14 days of the onset of symptoms. The relationship between delay and survival in bladder cancer is complex. Hospital delays may be influenced more by comorbidity than by the characteristics of the tumour. However, the adverse effects of delay seem to be most pronounced for patients with pT1 tumours.

Carbofuran toxicity on total lipids and free fatty acids in air breathing fish during exposure and cessation of exposure--in vivo.

Clarias batrachus, fish were exposed to sublethal concentration of commercial carbofuran for 144 hr. After 144 hr of exposure period, fish were transferred to fresh water and kept in the same for 144 hr in order to study the effect of carbofuran after cessation of intoxication. Total lipids and free fatty acids were determined in liver, muscle kidney and ovary at an interval of 24, 72 and 144 hr during exposure and after the release into toxicant free water. Total lipids increased in all the tissues throughout the exposure period, followed by decrease during depuration period. The results showed an alteration in free fatty acid content in different tissues of the fish. The elevated levels returned to almost control values after transfer of fish into carbofuran free water.

Age changes in some somatometric characters of the Assamese Muslims of Kamrup district, Assam.

Growth in weight, stature, sitting height, lower extremity length, head circumference and girth of biceps, chest and calf were studied in a cross-sectional sample of Assamese Muslims in Kamrup district, Assam, India. The sample comprised 835 boys and 894 girls, aged 3-20 years. The average growth pattern for the linear body dimensions was analysed by fitting Preece Baines model I to the mean values. The other dimensions were analysed in terms of simple line charts. Average final size and age at peak velocity of the linear dimensions were derived from the fitted curves. The data for height and weight were compared with other samples of the same district, with national urban and rural growth data for India, and with the NCHS growth standards. Mean height and weight of Assamese Muslim children fluctuates between the 5th and 10th centile of the NCHS standards up to approximately 15 years in boys and 13 years in girls. Thereafter, average length and weight lags behind the American standards, ending up below the 5th centile. The differences between the growth of Assamese Muslims and other populations are discussed.

Effect of acute exposure of the organophosphate insecticide Rogor on some biochemical aspects of Clarias batrachus (Linnaeus).

Indian catfish (Clarias batrachus), a common fish of commercial importance, was exposed to a sublethal concentration (1/3 of 96 h LC50 value) of Rogor for periods ranging from 24 to 192 h. The changes in glycogen, lactate, lactate dehydrogenase, and glycogen phosphorylase (a, ab) contents of muscle tissue were studied. A gradual decrease in muscle glycogen and an increase in lactate contents were observed. The activity level of lactate dehydrogenase showed a sharp rise initially for 48 h followed by a decline after 96 and 192 h. The glycogen phosphorylase a and ab in muscle tissue enhanced in response to Rogor exposure.

Carbohydrate metabolism in hepatic tissue of freshwater catfish Clarias batrachus L. during dimethoate exposure.

Clarias batrachus L., a freshwater catfish of commercial importance, was exposed to a sublethal concentration of dimethoate (Rogor). The acute effect of dimethoate on some aspects of carbohydrate metabolism of hepatic tissue was studied after 1, 2, 4 and 8 days. The glycogen content was depleted, whereas the lactate level was elevated throughout the experimental period and an increase in lactate dehydrogenase was observed. The activity levels of glycogen phosphorylase a and ab in the hepatic tissue were increased during dimethoate stress. The results suggest that carbohydrate metabolism was adversely affected in the hepatic tissue by the organophosphate insecticide dimethoate.