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Novel influenza A H1N1 virus (pH1N1) was the predominant strain in the 2009 pandemic. It continues to circulate along with other influenza strains in the post pandemic era. This new virus is nevertheless well known for its strong association with high cardiovascular mortality and a high propensity to preferentially affect young and otherwise healthy adults, resulting in a clustering of severe and even fatal cases in patients of the productive age group. H1N1 infection is a multifaceted disease. It affects the lung parenchyma as well as the pulmonary vasculature. It also affects the multisystemic vasculature and can cause other embolic events not strictly falling in the “usual” thrombotic category. We herein present a case of H1N1 influenza pneumonia induced severe ARDS with pulmonary thromboembolism (PTE) and pulmonary fibrosis.

Pulmonary arteriovenous malformations (PAVM) are rare anomalies of pulmonary vascular system which may be incidentally detected or can have manifestations resulting from right to left shunt. PAVM can cause various systemic implications including central nervous system complications like stroke, brain abscess due to paradoxical emboli which mostly seen in previously undiagnosed PAVMs indicating importance of early diagnosis and timely intervention in PAVM. Computed Tomography Pulmonary Angiography (CTPA) is the diagnostic method of choice in PAVM. We present here a unique case of complex multiple PAVM presented with refractory hypoxemia in a middle aged patient diagnosed in our centre initially evaluated for malignancy based on chest radiography & High Resolution Computed tomography(HRCT)thorax findings and symptoms, later on after CTPA was diagnosed to have complex PAVM which showed clinicoradiological improvement following pulmonary vascular plugging. This case shows clinical scenario, diagnostic & management methods and differential diagnosis to be discussed in similar clinicoradiological pattern and peculiarity of PAVM despite advanced age.

The aim of the current research is to assess the anticancer activity of the proteins identified from the crude protein extract derived from Hermetiaillucens larvae by computational methods. Bioinformatics methods were used to identify the protein sequence and virtual screening for the prediction of protein structure, their physico-chemical characteristics and functional aspects, that aid in exploring the anti-cancer interaction that is inclined to inhibit the oncoprotein activity. Due to the pathological process transpired by HPV, an array of genetic alterations, including the overexpression of oncoproteins or the inactivation of tumor suppressor proteins like TP53, BCL2, MDM2, ARF and BAX occur. The altered protein signaling pathways convulse to cervical cancer. In this research, we identified proteins, from the crude extract derived from Hermetiaillucens by LC-MS method. Further Computational screening of the selected proteins were employed to assess the functional units which has anticancer activity. We predicted four proteins Metallothionine, Defensin like precursor 1 protein, Heat shock protein 90 and NADH dehydrogenase and eluted the protein sequence from NCBI GenBank database. These sequences were used to predict physicochemical properties and protein structure prediction. Pharmacophore analysis of the peptide sequences as potential targets for cancer treatment was evaluated. Molecular docking of peptide sequences with target protein structure was carried out. To screen the best active potential molecule for cancer treatment the Molecular dynamics of docked protein-peptide structures were administered. The molecular docking of the peptides with onco-proteins has been predicted and virtually screened based on RMSD values. The resultant protein sequence was evaluated further by amino acid sequencing, extensive scrutiny of the proteins, advanced computational imaging, molecular docking examination and assessed the results for clinical interpretation. To summate, bioinformatics assessment enabled us to identify four key proteins derived from Hermetiaillucens larvae that have strong inhibitory function against proteins that cause cervical cancer.

Background: Sleep-disordered breathing (SDB), predominantly obstructive sleep apnea (OSA), is a frequent phenomenon in interstitial lung disease (ILD) and may be associated with significant morbidity and mortality. Methodology: A prospective, observational, hospital-based study was conducted in a tertiary care hospital after ethics committee permission. The study group consisted of 100 consecutive ILD patients diagnosed by a multidisciplinary diagnosis. They were evaluated for the prevalence of SDB with a polysomnography after a comprehensive history, detailed clinical examination, calculation of various pretest probability scores, and relevant prerequisite workup. Results: Out of the total 100 ILD patients, 44 were male (44%) and 56 were female (56%). SDB was present in 57 (57%) patients. Of these, 29 (29%) were found to have only nocturnal oxygen desaturation (NOD), while 28 (28%) had OSA. The 28 cases of OSA were distributed as 15 mild OSA (53.57%), 10 moderate OSA (35.71%), and 3 severe OSA (10.71%). The patients were divided into the following four groups: total study Group (A), patients with OSA (Group B), patients with NOD without OSA (Group C), and no SDB (Group D). The mean forced vital capacity values predicted in the four groups were 53.67%, 50%, 45.56%, and 57.87%, respectively. The mean body mass index in the four groups was 24.56, 27, 26.98, and 24.89 kg/m2, respectively. The mean 6-min walk distance in the four groups was 280.7, 250, 256.65, and 311.4 m, respectively. The mean partial pressure of oxygen in the four groups was 65.65, 60, 62.10, and 75.66 mmHg, respectively. The mean apnea–hypopnea index in the study group was 2.98/h, 8.6/h with mild OSA, 21.69/h with moderate OSA, 48.78/h with severe OSA, 3.89/h in patients having NOD without OSA, and 2.54/h in patients with no SDB. Conclusion: SDB in ILD is associated with a significant impact on the cardinal determinants of functional capacity, lung function, and quality of life.

Lay summary The current pandemic of COVID-19 has affected all the countries globally. However, the adverse impact of the pandemic is more seen in the low-income and middle-income countries (LMICs). Although there is evidence on the adverse impact of the SARS-CoV-2 on the health of mothers and neonates, the evidence is mainly from high-income countries. For reducing the mortality and morbidity due to COVID-19 in LMICs, there is a need to generate evidence from the LMICs. The present study is a part of the National Registry of pregnant women with COVID-19 in India (PregCovid registry). Our study demonstrates a higher risk of adverse outcomes such as neonatal sepsis and death in the SARS-CoV-2 infected as compared to the non-infected neonates. The study also showed the risk of SARS-CoV-2 infection in 6.3% of neonates born to mothers with COVID-19. Introduction We describe the clinical characteristics, management, and short-term outcomes of SARS-CoV-2 neonates born to mothers with COVID-19 in a tertiary care hospital in Mumbai, India. Methods The study is a retrospective analysis of 524 neonates born to mothers with COVID-19 admitted from 14th April 2020 to 31st July 2020. Results SARS-CoV-2 infection was detected in 6.3% of the newborns of the mothers with COVID-19. No significant differences were observed between maturity at gestation, birth weight and sex of SARS-CoV-2 infected and non-infected newborns. The risk of sepsis was 4.09 (95% CI,1.28-13.00) fold higher in the neonates with SARS-CoV-2 as compared to the non-infected group (p = 0.031). Poor feeding was significantly more common among SARS-CoV-2 infected neonates (12.1%) as compared to the non-infected neonates (2.7%) (p = 0.017). There was a total of 13 neonatal deaths, of which 3 deaths occurred in SARS-CoV-2 infected neonates (9%) while 10 (3%) in the SAR-CoV-2 negative group. The risk of neonatal death was higher in SARS-CoV-2 infected newborns (OR 4.8; 95% CI 1.25-18.36). Conclusion Neonatal SARS-CoV-2 infection is observed in almost 6% of neonates born to mothers with perinatal COVID-19. There is a higher risk of adverse outcomes such as neonatal sepsis and death in the SARS-CoV-2 infected as compared to the non-infected neonates.

Background: Acute phase insulin secretion is disturbed in individuals with prediabetes. We have previously shown a significant risk reduction in 4,417 prediabetes individuals through Yoga. The present study explores yoga’s mechanism of action. Method: This randomized control trial included 42 (20 yoga;22 walkings;5 days/ week, 3Months) prediabetes (fasting glucose of 100 to 125 mg/dl) adults between 23 to 50 years. Serum Insulin and C peptide were assessed at 0, 3, 5, 7, 10, 15, 30,60and 90th minutes after 75grams of oral glucose. Pre-post assessments included fasting Glucose, HOMA%B(derived), GLP-1, free Triiodothyronine, perceived stress, and Heart rate variability. Results: There was a significant increase in the 10th minute (acute phase) Insulin and c-peptide levels in the yoga as compared to the control group. Improvement in GLP-1 and HOMA%B was significantly better in yoga than walking group. Yoga group also showed positive change in perceived stress and HRV. Simple linear regression of post data using acute phase Insulin as the dependent variable indicated a significant association between all measures(R2 = 0.806; p = Conclusion: The shift to normoglycemia, due to an increase in acute phase insulin release, was associated with improved autonomic balance and reduced stress after yoga. More studies may confirm this unique first time observation. Disclosure R. Nagarathna: None. J. Kurian: None. V. Vijayakumar: None. H. R. Nagendra: None. R. N. Mavathur: None. Funding Ministry of AYUSH, Government of India (Z.28015-HPC (EMR)-AYUSH-B)

ObjectiveTo understand the outcome of hospitalised patients from Mumbai City, which had the highest number of COVID-19 cases in India.DesignObservational study with follow-up.SettingData extraction from medical records of patients with COVID-19 admitted to Nair Hospital & TN Medical College, Mumbai, India.Participants689 patients with COVID-19 were admitted in the hospital from 26 March 2020 to 11 May 2020.Primary and secondary outcome measuresIn-hospital mortality; joint effect of comorbidity and age on the risk of dying.ResultsA total of 689 patients (median age 44 years) admitted with RT-PCR-confirmed COVID-19 were included in the study. Of these, 77.36% of patients were discharged alive while 22.64% died. 11.61% required some kind of oxygen support while 2.8% of patients required intensive care unit admissions. Older age (HR 2.88, 95% CI 2.09 to 3.98), presence of comorbidities (HR 2.56, 95% CI 1.84 to 3.55), history of hypertension (HR 3.19, 95% CI 1.67 to 6.08), and presence of symptoms at the time of admission (HR 3.21, 95% CI 1.41 to 7.26) were associated with increased risk of in-hospital mortality. Treatment with a combination of azithromycin with hydroxychloroquine, antiviral or steroid compared with no treatment did not alter the disease course and in-hospital mortality. The combined effect of old age and presence of comorbid conditions was more pronounced in women than men.ConclusionsIn-hospital patients were younger, less symptomatic with lesser need of ventilators and oxygen support as compared with many western countries.Trial registrationNot applicable (observational study, not a clinical trial).

Background: Coronavirus disease (COVID-19) is an infectious disease caused by SARS-CoV-2, clinically presenting with common symptoms of fever, dry cough, and breathlessness within 14 days of exposure. Its severity ranges from mild to severe, latter manifesting into severe acute respiratory syndrome. As a part of multidisciplinary team, physiotherapy along with medical management was administered to patients with COVID-19 in an acute care setup. This retrospective study aims to explore various patient characteristics and will aid in identifying the impairments associated with the disease, giving a direction to the physiotherapy community in planning future management strategy to improve quality of life. Patients and methods: The present study is a unicentric study wherein prospective analysis of retrospective data of patients referred for physiotherapy from May 13 to July 31, 2020, was performed. (i) Characteristics of patients, (ii) associated comorbidities, (iii) hospital course since the time of admission to discharge, (iv) mode of oxygen delivery, (v) pre- and post-physiotherapy treatment values of oxygen saturation and heart rate, and (vi) physiotherapy treatment were recorded. The archived data were analyzed using the commercially available SPSS software version 24. Wilcoxon's matched pair test was used to compare pre- and post-treatment oxygen saturation and heart rate, and McNemar's test was used to compare mode of oxygen delivery and pre- and post-physiotherapy treatment. Results: Descriptive analysis of data showed a better outcome in terms of grade of dyspnea and rate of discharge on day 14 of physiotherapy treatment. Hence, a comparative analysis of day 1 and day 14 was performed for mode of oxygen delivery, oxygen saturation, and heart rate. A statistically significant improvement was observed in the heart rate (p = 0.001) and oxygen delivery (p = 0.000). However, no significant difference in the level of oxygen saturation was found (p = 0.6433). Conclusion: Physiotherapy treatment in conjunction with medical treatment can be effectively administered in patients with COVID-19 in acute care setup taking into consideration the health status and the hemodynamic stability of the patients. It emphasizes the role of physiotherapy in the alleviation of symptoms, facilitating early weaning and recovery enabling early discharge from the hospital. How to cite this article: Verma CV, Arora RD, Mistry HM, Kubal SV, Kolwankar NS, Patil PC, et al. Changes in Mode of Oxygen Delivery and Physiological Parameters with Physiotherapy in COVID-19 Patients: A Retrospective Study. Indian J Crit Care Med 2021;25(3):317–321.

BACKGROUND A respiratory intensive care unit (RICU) is a designated area specially designed for management of patients with acute respiratory conditions with an essential endeavour of meticulous cardiorespiratory monitoring and holistic management of acute respiratory conditions. Availability of ventilation thorough the non-invasive mode (NIV) has revolutionised therapy in chronic respiratory diseases. METHODS We analysed the data of 50 patients over two-months’ duration. The mean duration of stay and treatment outcomes were analysed. The qualitative data was analysed using percentages and mean. Chi-Square test was used for statistical analysis. RESULTS Study sample was of 50 patients out of which 26 were men and 24 were women. The indications for hospitalisation were classified as per aetiology as COPD- 29 (58%), ILD- 6 (12%), parenchymal diseases- 5 (10%), pleural diseases- 4 (8%), malignancy- 3 (6%), others- 3 (6%). The patients were further classified into; 21 (42%) with type I respiratory failure and 29 (58%) with type II respiratory failure. Of the total 50; one-third (18) required only oxygen therapy and two-third (32) mechanical ventilation (25 NIV and 7 IMV). The mode of ventilation impacted outcomes significantly with oxygen therapy and NIV having very good outcomes (Chi Square test; p-value is 0.000838). CONCLUSIONS RICU is the need of the hour to manage patients with critical respiratory illnesses.

Immune cell dysregulation and lymphopenia characterize COVID-19 pathology in moderate to severe disease. While underlying inflammatory factors have been extensively studied, homeostatic and mucosal migratory signatures remain largely unexplored as causative factors. In this study we evaluated the association of circulating IL-6, soluble mucosal addressin cell adhesion molecule (sMAdCAM) and IL-15 with cellular dysfunction characterizing mild and hypoxemic stages of COVID-19. A cohort of SARS-CoV-2 infected individuals (n=125) at various stages of disease progression together with healthy controls (n=16) were recruited from COVID Care Centres (CCCs) across Mumbai, India. Multiparametric flow cytometry was used to perform in-depth immune subset characterization and to measure plasma IL-6 levels. sMAdCAM, IL-15 levels were quantified using ELISA. Distinct depletion profiles, with relative sparing of CD8 effector memory and CD4+ regulatory T cells was observed in hypoxemic disease within the lymphocyte compartment. An apparent increase in the frequency of intermediate monocytes characterized both mild as well as hypoxemic disease. IL-6 levels inversely correlated with those of sMAdCAM and both markers showed converse associations with observed lympho-depletion suggesting opposing roles in pathogenesis. Interestingly, IL-15, a key cytokine involved in lymphocyte activation and homeostasis, was detected in symptomatic individuals but not in healthy controls or asymptomatic cases. Further, negative association of plasma IL-15 with depleted T, B and NK subsets suggested a compensatory production of this cytokine in response to the profound lymphopenia. Finally, higher levels of plasma IL-15 and IL-6, but not sMAdCAM, were associated with longer duration of hospitalization.

Awareness of breast cancer has been increasing due to early detection, but the advanced disease has limited treatment options. There has been growing evidence on the role of miRNAs involved in regulating the resistance in several cancers. We performed a comprehensive systematic review and meta-analysis on the role of miRNAs in influencing the chemoresistance and sensitivity of breast cancer. A bibliographic search was performed in PubMed and Science Direct based on the search strategy, and studies published until December 2018 were retrieved. The eligible studies were included based on the selection criteria, and a detailed systematic review and meta-analysis were performed based on PRISMA guidelines. A random-effects model was utilised to evaluate the combined effect size of the obtained hazard ratio and 95% confidence intervals from the eligible studies. Publication bias was assessed with Cochran’s Q test, I2 statistic, Orwin and Classic fail-safe N test, Begg and Mazumdar rank correlation test, Duval and Tweedie trim and fill calculation and the Egger’s bias indicator. A total of 4584 potential studies were screened. Of these, 85 articles were eligible for our systematic review and meta-analysis. In the 85 studies, 188 different miRNAs were studied, of which 96 were upregulated, 87 were downregulated and 5 were not involved in regulation. Overall, 24 drugs were used for treatment, with doxorubicin being prominently reported in 15 studies followed by Paclitaxel in 11 studies, and 5 drugs were used in combinations. We found only two significant HR values from the studies (miR-125b and miR-4443) and our meta-analysis results yielded a combined HR value of 0.748 with a 95% confidence interval of 0.508−1.100; p-value of 0.140. In conclusion, our results suggest there are different miRNAs involved in the regulation of chemoresistance through diverse drug genetic targets. These biomarkers play a crucial role in guiding the effective diagnostic and prognostic efficiency of breast cancer. The screening of miRNAs as a theragnostic biomarker must be brought into regular practice for all diseases. We anticipate that our study serves as a reference in framing future studies and clinical trials for utilising miRNAs and their respective drug targets.

Bioinformatics is the latest technology in the research field. In-silico studies are done to identify the exact target of the drug. Which finds a drug for the particular binding site and final stage animal testing can be done for obtaining a conform result. Reviews of in-silico studies in herbal drugs were retrieved through the use of PubMed. Specific software on a computer allows researchers to analyse enormous data without actually conducting a large number of experiments. It helps to give the existing information to model disease pathway and identifies precise targets of the selected drugs. Modern instrumental techniques like XRD, XRF, ICP-MS etc. help in quantitative and qualitative estimation of metals and minerals and structural analysis of compound drugs. A later stage in vivo and in vitro studies can be done for obtaining the confirmatory result.

INTRODUCTION: Esophageal carcinoma is one of the most challenging problems confronting the surgeons. Majority of the patients present late. In developing countries like ours, chewing of tobacco leaves along with use of betel nuts and leaves also, in addition to smoking and alcoholism, have been implicated in the development of esophageal carcinoma. Surgery is still the initial treatment of choice for a curative intent. Since the advancement of investigations, we can now detect these cancers at an early stage thereby improving the long term relief following surgery. MATERIALS AND METHODS: This study was conducted in patients who underwent surgery for malignant esophageal tumours in the Departments of Surgery and Surgical Gastroenterology, from September 2009 to March 2011. All the patients who were diagnosed and operated for carcinoma esophagus were included in the study. Data collected was analyzed and studied for understanding the pattern of the incidence, disease presentation, age/sex distribution, etiological factors, clinicopathological characteristics, complications and outcome of surgery. RESULTS: A total of 30 patients were operated with M:F ratio 2:1. Carcinoma esophagus showed peak age incidence in the 5th-6th decade. More than 90% belonged to the poor socioeconomic class. Nearly 75-80% of patients were either smokers, alcoholics or were addicted to betel nut chewing. Dysphagia was the primary mode of presentation. Esophagogastroduodenoscopy was done in all patients. Lesions were predominantly seen in lower 1/3rd of the esophagus. Histopathologically, 22(73.3%) had squamous cell carcinoma while the remaining 8(26.7%) had adenocarcinoma. All 30 patients were operated and of these, 26(86.6%) patients underwent trans-hiatal esophagectomy and rest underwent thoraco-laparoscopic procedure. 1(3.3%) patient had splenic tear intra-operative for which splenectomy was performed. 4(13.3%) patients developed complications like minor cervical anastomosis leak in 2(6.6%) patients, pleural effusion 1(3.3%) and pneumonia 1(3.3%), which were managed conservatively. There were no stricture/stenosis of the anastomotic site and no deaths occurred in the postoperative period. CONCLUSION: Squamous cell carcinoma of the esophagus is seen more than adenocarcinoma. Tobacco use and alcohol consumption along with presence of preexisting esophageal conditions are strong risk factors for the development of the disease. Disease shows predominance to male and is more common in 5th-6th decade of life. Carcinoma of lower third of esophagus is more common. Surgical approaches do not alter the mortality or survival although duration of stay may be reduced.

Biocatalytic processes to selectively hydrolyze the N-carbobenzyloxy (CBz) group from CBz-protected d - or l -amino acids have been developed. The substrate specificities of the CBz-deprotecting enzymes from Burkholderia phenazinium SC 16530, and Sphingomonas paucimobilis expressed into Escherichia coli SC 16501 were evaluated on CBz-protected amino acids and structurally related compounds. Modifications of various structural components and their effects on enzyme activity and enantioselectivity provided a greater understanding of the two CBz-deprotecting enzymes.

Background: Sacroiliac joints (SJ) involvement is a distinctive and charasteristic feature of Spondyloarthritis (SpA) and x-ray is the test routinely used to make a diagnosis. However, x-ray reveals late structural damage but cannot detect active inflammation. The objective of this study was to assess the validity of Doppler ultrasound in SJ. Methods: Prospective blinded and controlled study of SJ, in which three populations were compared. We studied 106 consecutive cases, who were divided into three groups: a) 53 patients diagnosed with SpA who had inflammatory lumbar and gluteal pain assessed by a rheumatologist; b) 26 patients diagnosed with SpA who didn't have SJ tenderness and had normal physical examination; c) control group of 27 subjects (healthy subjetcs or with mechanical lumbar pain). All patients included that were diagnosed with SpA met almost the European Spondyloarthropathy Study Group (ESSG) classification criteria. Physical examination of the SJ included: sacral sulcus tenderness, iliac gapping, iliac compression, midline sacral thrust test, Gaenslen's test, and Patrick s test were used as gold standard. Both SJ were examined with Doppler ultrasound (General Electric Logiq 9, Wauwatosa WI, USA) fitted with a 9-14 Mhz lineal probe. The ultrasonographer was blinded to clinical data. Doppler in SJ was assessed as positive when both Doppler colour and resistance index (RI)

A synthetic gene encoding a Streptomyces l -proline-3-hydroxylase was constructed and used to produce the hydroxylase protein in recombinant Escherichia coli. A fermentation process for growth of this recombinant E. coli for enzyme production was scaled-up to 250 L. A biotransformation process was developed using cell suspensions of the recombinant E. coli and subsequently scaled-up to 10 L for conversion of l -proline to cis-3-hydroxy- l -proline. A reaction yield of 85 M% and d.e. of 99.9% was obtained for cis-3-hydroxy- l -proline.

Calcineurin is an important signalling protein that regulates a number of molecular and cellular processes. Previously, we found that inhibition of calcineurin with cyclosporine reduced renal hypertrophy and blocked glomerular matrix expansion in the diabetic kidney. Isoforms of the catalytic subunit of calcineurin are reported to have tissue specific expression and functions. In particular, the β isoform has been implicated in cardiac and skeletal muscle hypertrophy. Therefore, we examined the role of calcineurin β in diabetic renal hypertrophy and glomerular matrix expansion. Type I diabetes was induced in wild-type and β(-/-) mice and then renal function, extracellular matrix expansion and hypertrophy were evaluated. The absence of β produced a significant decrease in total calcineurin activity in the inner medulla (IM) and reduced nuclear factor of activated T-cells (NFATc) activity. Loss of β did not alter diabetic renal dysfunction assessed by glomerular filtration rate, urine albumin excretion and blood urea nitrogen. Similarly, matrix expansion in the whole kidney and glomerulus was not different between diabetic wild-type and β(-/-) mice. In contrast, whole kidney and glomerular hypertrophy were significantly reduced in diabetic β(-/-) mice. Moreover, β(-/-) renal fibroblasts demonstrated impaired phosphorylation of Erk1/Erk2, c-Jun N-terminal kinases (JNK) and mammalian target of rapamycin (mTOR) following stimulation with transforming growth factor-β and did not undergo hypertrophy with 48 hrs culture in high glucose. In conclusion, loss of the β isoform of calcineurin is sufficient to reproduce beneficial aspects of cyclosporine on diabetic renal hypertrophy but not matrix expansion. Therefore, while multiple signals appear to regulate matrix, calcineurin β appears to be a central mechanism involved in organ hypertrophy.

Calcineurin is an important intracellular signaling molecule which can be inhibited by cyclosporin resulting in immune suppression and nephrotoxicity. Previously, we reported that homozygous loss of the α isoform of calcineurin impairs kidney development and function and mimics many features of cyclosporin nephrotoxicity. However, early lethality of null mice prevented further study of renal changes. Alternatively, we examined aged heterozygous (CnAα+/−) mice. In addition to renal dysfunction and inflammation, we find that CnAα+/− mice spontaneously develop tertiary lymphoid aggregates in the kidney, small intestine, liver, and lung. Lymphoid aggregates contain both T cells and B cells and exhibited organization suggestive of tertiary lymphoid organs (TLOs). Kidney function and TLO formation were highly correlated suggesting that this process may contribute to nephrotoxicity. Consistent with previous findings, transforming growth factor (TGF)-β is significantly increased in CnAα+/− mice. Neutralization of TGF-β attenuated TLO formation and improved kidney function. In conclusion, we report that haploinsufficiency of CnAα causes uregulation of TGF-β which contributes to chronic inflammation and formation of TLOs. While the process that leads to TLOs formation in transplant allografts is unknown, TLOs are associated with poor clinical prognosis. This study suggests that calcineurin inhibition itself may lead to TLO formation and that TGF-β may be a novel therapeutic target.

Previously we have demonstrated the reduction of ethyl and t-butyl diketoesters 1 to the corresponding syn-(3R,5S)-dihydroxy esters 2a by Acinetobacter sp. 13874. The syn-(3R,5S)-dihydroxy ester 2a was obtained with an enantiomeric excess (e.e.) of 99% and a diastereomeric excess (de) of 63%. In this report, we identified a gene encoding desired ketoreductase III which catalyzed the diastereoselective reduction of diketoesters 1 to syn-(3R,5S)-dihydroxy esters 2a and describe cloning and expression of ketoreductase III into Escherichia coli. Cells or extracts of recombinant E. coli efficiently reduced the diketoester 1 to the corresponding syn-(3R,5S)-dihydroxy ester 2a in 99.3% yield, 100% e.e., and 99.8% de.

The d-amino acid (R)-2-amino-3-(7-methyl-1 H-indazol-5-yl)propanoic acid (3) is a key intermediate needed for synthesis of a drug candidate compound. Enzymatic routes to 3 were explored. d-Amino acid 3 was prepared in 68% isolated yield with >99% ee from racemic amino acid 1 using l-amino acid deaminase from Proteus mirabilis expressed in Escherichia coli in combination with a commercially available d-transaminase using d-alanine as amino donor. The d-enantiomer was also prepared in 79% isolated yield with >99% ee from the corresponding keto acid 2 using the d-transaminase with racemic alanine as the amino donor. The rate and yield of this reaction could be accelerated by addition of lactate dehydrogenase (with NAD, formate and formate dehydrogenase to regenerate NADH) to remove the inhibitory pyruvate produced during the reaction. A d-transaminase was purified from a soil organism identified as Bacillus thuringiensis and cloned and expressed in E. coli. The d-transaminase was very effective for the prepa...

Protection against a high dose of a toxicant by prior exposure to another toxicant is called heteroprotection. Our objective was to establish a heteroprotection model in RBCs. Female Sprague Dawley rats treated with an LD90 dose of 2-butoxyethanol (BE, 1500 mg/kg in water, 5 ml/kg po) 14 days after priming with 0.9% NaCl suffered 90% mortality by 15 days, whereas all rats receiving the LD90 dose of BE 14 days after priming with phenylhydrazine (PHZ, 125 mg/kg in 0.9% NaCl, 3 ml/kg po) survived. Hematocrit decreased from normal 45% to 24% by day 3 after PHZ priming and improved thereafter. Increasing the time interval between the priming and LD90 dose to 21 days abolished the heteroprotection. RBCs obtained on days 7 and 14 after PHZ priming unlike those on day 21 were resilient to the hemotoxic metabolite of BE, butoxyacetic acid (BAA). Unaltered hepatic alcohol and aldehyde dehydrogenase activities upon PHZ priming suggested that bioactivation of BE to BAA was unaffected. Lower renal (6 and 12 h) and hepatic (12 h) BAA levels and 3 fold higher excretion of BAA in PHZ-primed rat urine suggested a protective role of toxicokinetics. Higher erythropoietin, reticulocytes, and resiliency of PHZ-primed rat RBCs indicated that newly formed RBCs are resilient to hemolytic BAA. The antioxidant levels in the PHZ-primed rat RBCs did not indicate a protective role in heteroprotection. In conclusion, the resistance of PHZ-primed rats against BE-induced hemotoxicity and lethality is mediated by a combination of altered toxicokinetics, robust erythropoiesis, and resiliency of new RBCs.

The enantioselective microbial reduction of 6-oxo-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-dione (1) to either of the corresponding (S)- and (R)-6-hydroxy-8-[4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl]-8-azaspiro[4.5]decane-7,9-diones (2 and 3, respectively) is described. The NADP+-dependent (R)-reductase (RHBR) which catalyzes the reduction of 6-ketobuspirone (1) to (R)-6-hydroxybuspirone (3) was purified to homogeneity from cell extracts of Hansenula polymorpha SC 13845. The subunit molecular weight of the enzyme is 35,000 kDa based on sodium dodecyl sulfate gel electrophoresis and the molecular weight of the enzyme is 37,000 kDa as estimated by gel filtration chromatography. (R)-reductase from H. polymorpha was cloned and expressed in Escherichia coli. To regenerate the cofactor NADPH required for reduction we have cloned and expressed the glucose-6-phosphate dehydrogenase gene from Saccharomyces cerevisiae in E. coli. The NAD+-dependent (S)-reductase (SHBR) which catalyzes the reduction of 6-ketobuspirone (1) to (S)-6-hydroxybuspirone (2) was purified to homogeneity from cell extracts of Pseudomonas putida SC 16269. The subunit molecular weight of the enzyme is 25,000 kDa based on sodium dodecyl sulfate gel electrophoresis. The (S)-reductase from P. putida was cloned and expressed in E. coli. To regenerate the cofactor NADH required for reduction we have cloned and expressed the formate dehydrogenase gene from Pichia pastoris in E. coli. Recombinant E. coli expressing (S)-reductase and (R)-reductase catalyzed the reduction of 1 to (S)-6-hyroxybuspirone (2) and (R)-6-hyroxybuspirone (3), respectively, in >98% yield and >99.9% e.e.

Objective of the present study was to test the importance of tissue repair in the final outcome of S-(1,2-dichlorovinyl)-L-cysteine (DCVC)-induced nephrotoxicity using colchicine (CLC) intervention. Male Swiss Webster (SW) mice were administered a normally nonlethal dose of DCVC (30 mg/kg, i.p.) on day 0 and CLC (2 mg/kg, i.p.) at 42 and 66 h after administration of DCVC. The mice were observed for mortality and various renal injury and repair parameters were studied during a time course of 0-14 days. Administration of 30 mg DCVC/kg led to loss of renal architecture by day 1, which sustained until day 5, and regressed thereafter to reach normal architecture by day 10 resulting in 100% survival. Renal dysfunction as assessed by increases in plasma BUN and creatinine levels was concordant during this time course. Urinary volume increased significantly between days 10 and 14 with significant increases in urinary glucose concentrations on days 1-4. Calpain leakage increased from day 1 and remained so until day 5 before declining at later time points. In contrast, CLC intervention led to marked inhibition of S-phase DNA synthesis and 100% mortality by 120 h. H&E sections of kidneys revealed loss of renal architecture on day 1 which progressively worsened from day 2 to 4. Polyuria and glycosuria were evident during the first 2 and 3 days, respectively. Calpain immunohistochemistry revealed progressive leakage of calpain in the extracellular space during 2-4 days which lead to increased renal injury as evident from significant increases in calpain specific breakdown products (CSBPs) of alpha-fodrin during the same period of time. The group of mice receiving 2 mg CLC/kg alone showed a significant increase in urinary creatinine concentration on day 5. Neither the expression nor localization of aquaporin 1 was altered in any of the treatment groups. These results show that antimitotic intervention after DCVC-initiated renal injury leads to expansion and progression of that injury, which appears to be due to proteolytic destruction of neighboring cells mediated by calpain leaking out of necrosed renal tubular epithelial cells.

A novel enantioselective N-carbobenzyloxy (Cbz)-cleaving enzyme was purified to homogeneity from Sphingomonas paucimobilis SC 16113. The gene encoding the protein has been identified and its complete DNA sequence was determined. The Cbz-cleaving enzyme was expressed in Escherichia coli.

A new enzymatic process for the enantioselective cleavage of N-benzyloxycarbonyl (Cbz) groups from protected amino acids and related compounds has been developed. The Cbz-deprotecting enzyme was isolated from cell extracts of Sphingomonas paucimobilis SC 16113 and purified to homogeneity. The purified protein has a molecular weight of 155,000 daltons and a subunit size of 44,000 daltons.

Nosocomial pneumonia poses great challenge to an intensivist. Detailed information about hospital-acquired pneumonia (HAP) and ventilator-acquired pneumonia (VAP) is crucial for prevention and optimal management, thus improving quality Intensive Care Unit (ICU) care. Hence, we aimed to study the current trend of nosocomial pneumonia in ICU.It was a prospective observational cohort study, conducted in the ICU of a tertiary care teaching public hospital over a period of 18 months. We studied clinical profile and outcome of 120 adult patients who developed VAP/HAP during the study period. We also analyzed the causative organisms, antibiotic sensitivity, and resistance pattern in these patients.Out of 120 patients, 29 patients were HAP and 91 patients were VAP. Mortality was 60% (72), and development of VAP and requirement of mechanical ventilation showed significant association with mortality (P0.00001). Most common organism causing HAP was Staphylococcus aureus (43.4%) and VAP was Klebsiella pneumoniae (49%). Maximum antibiotic sensitivity was found to piperacillin + tazobactam (58.8%), followed by imipenem (49.5%) and meropenem (41.8%), whereas maximum antibiotic resistance was found to cefepime (95.1%), followed by ceftazidime and amoxicillin (91.2%).Nosocomial pneumonia showed high incidence (17.44%) and mortality (60%). Common organisms identified were S. aureus and K. pneumoniae. Resistance was high for commonly used antibiotics and high antibiotic sensitivity for piperacillin + tazobactam and carbapenem. Contexte: La pneumonie nosocomiale pose un grand défi à un intensiviste. Des informations détaillées sur la pneumonie acquise dans les hôpitaux (HAP) et la pneumonie acquise par le ventilateur (VAP) sont essentielles pour la prévention et la gestion optimale, améliorant ainsi les soins de soins intensifs de qualité (UTI). Par conséquent, nous avons cherché à étudier la tendance actuelle de la pneumonie nosocomiale en UTI. Matériaux et méthodes: il s'agissait d'une étude de cohorte observationnelle prospective menée dans l'UCI d'un hôpital public d'enseignement tertiaire sur une période de 18 mois. Nous avons étudié le profil clinique et le résultat de 120 patients adultes qui ont développé le VAP / HAP pendant la période d'étude. Nous avons également analysé les organismes responsables, la sensibilité aux antibiotiques et le modèle de résistance chez ces patients. Résultats: Sur 120 patients, 29 patients étaient HAP et 91 patients étaient VAP. La mortalité était de 60% (72), et le développement du VAP et l'exigence de ventilation mécanique ont montré une association significative avec la mortalité (P0,00001). L'organisme le plus fréquent causant HAP était Staphylococcus aureus (43,4%) et VAP était Klebsiella pneumoniae (49%). Une sensibilité antibiotique maximale a été observée chez la pipéracilline + tazobactam (58,8%), suivie de l'imipénème (49,5%) et du méropénem (41,8%), alors que la résistance antibiotique maximale a été observée à cefépime (95,1%), suivie de ceftazidime et de l'amoxicilline (91,2%) .la pneumonie nosocomiale a montré une incidence élevée (17,44%) et la mortalité (60%). Les organismes communs identifiés étaient S. aureus et K. pneumoniae. La résistance était élevée pour les antibiotiques couramment utilisés et une forte sensibilité aux antibiotiques pour la pipéracilline + le tazobactam et le carbapénème.

CETP activity, measured as transfer of cholesteryl ester from exogenous HDL to exogenous VLDL and LDL, reflecting CETP mass as determined by ELISA, was documented in three groups of St. Kitts vervet monkeys fed diets enriched in saturated (Sat), monounsaturated (Mono), or n-6 polyunsaturated (Poly) fatty acids. CETP activity was not different when comparing the three dietary fats. However, CETP activity was significantly higher when cholesterol was added to each of the diets. Significant positive associations between CETP activity and VLDL and LDL cholesterol concentrations were found whereas significant negative associations were seen between CETP activity and HDL cholesterol in each of the diet groups. The strength of these associations was highest in the Sat group. Cholesteryl ester (CE) fatty acid composition of lipoproteins varied widely among diet groups, with the more polyunsaturated CE of the Poly group being associated with a higher rate of CE transfer to endogenous acceptor apolipoprotein B-containing lipoproteins. Finally, only the Sat diet group showed significant positive correlations of CETP activity with LDL particle diameter (r = 0.76), cholesteryl ester percentage (r = 0.67), and a strong negative correlation (r = −0.86) with LDL receptor function, estimated as the difference between native and methylated LDL turnover rates. We speculate that strong associations between CETP and LDL metabolism may explain, at least in part, the increased atherogenicity of dietary saturated fat. —Fusegawa, Y., K. L. Kelley, J. K. Sawyer, R. N. Shah, and L. L. Rudel. Influence of dietary fatty acid composition on the relationship between CETP activity and plasma lipoproteins in monkeys.

Mice lacking the α isoform of the catalytic subunit of calcineurin (CnAα) were first reported in 1996 and have been an important model to understand the role of calcineurin in the brain, immune system, bones, muscle, and kidney. Research using the mice has been limited, however, by failure to thrive and early lethality of most null pups. Work in our laboratory led to the rescue of CnAα-/- mice by supplemental feeding to compensate for a defect in salivary enzyme secretion. The data revealed that, without intervention, knockout mice suffer from severe caloric restriction. Since nutritional deprivation is known to significantly alter development, it is imperative that previous conclusions based on CnAα-/- mice are revisited to determine which aspects of the phenotype were attributable to caloric restriction versus a direct role for CnAα. In this study, we find that defects in renal development and function persist in adult CnAα-/- mice including a significant decrease in glomerular filtration rate and an increase in blood urea nitrogen levels. These data indicate that impaired renal development we previously reported was not due to caloric restriction but rather a specific role for CnAα in renal development and function. In contrast, we find that rather than being hypoglycemic, rescued mice are mildly hyperglycemic and insulin resistant. Examination of muscle fiber types shows that previously reported reductions in type I muscle fibers are no longer evident in rescued null mice. Rather, loss of CnAα likely alters insulin response due to a reduction in insulin receptor substrate-2 (IRS2) expression and signaling in muscle. This study illustrates the importance of re-examining the phenotypes of CnAα-/- mice and the advances that are now possible with the use of adult, rescued knockout animals.

Chiral intermediates were prepared by biocatalytic processes for the chemical synthesis of three pharmaceutical drug candidates. These include (i) the synthesis of [(3R-cis)-3-(acetyloxy)-4-phenyl-2-azetidinone2 for the semi-synthesis of paclitaxel (taxol)5, an anticancer compound; (ii) synthesis of chiral (exo,exo)-7-oxabicyclo [2.2.1] heptane-2,3-dimenthanol monoacetate ester9 for the chemoenzymatic preparation of a thromboxane A2 antagonist; (iii) the enzymatic synthesis ofS-(−) 3-benzylthio-2-methylpropanoic acid, a key chiral intermediate for the synthesis of antihypertensive drugs captopril10 or zofenopril13.

Acute myocardial infarction (AMI) is the leading cause of death worldwide, with early diagnosis still being difficult. Promising new cardiac biomarkers such as troponins and creatine kinase (CK) isoforms are being studied and integrated into clinical practice for early diagnosis of AMI. The cardiac-specific troponins I and T (cTnI and cTnT) have good sensitivity and specificity as indicators of myocardial necrosis and are superior to CK and its MB isoenzyme (CK-MB) in this regard. Besides being potential biologic markers, cardiac troponins also provide significant prognostic information. The introduction of novel high-sensitivity troponin assays has enabled more sensitive and timely diagnosis or exclusion of acute coronary syndromes. This review summarizes the available information on the potential of troponins and other cardiac markers in early diagnosis and prognosis of AMI, and provides perspectives on future diagnostic approaches to AMI.

10-Deacetylabaccatin III (10 DAB), an important precursor for paclitaxel semisynthesis, is enhanced in yew extracts using C10-deacetylase and C13-deacylase enzymes.(4) C10-deacetylase is an intracellular enzyme produced by the fermentation of a soil microorganism, Nocardioides luteus (SC 13912). During the fermentation of Nocardioides luteus, the growth of cells reaches a maximum growth at 28 h. C10-deacetylase enzyme activity starts at 26 h and peaks at 38 h of the fermentation. The cells are recovered by centrifugation. The C10-deacetylase enzyme was purified from the Nocardioides luteus cells. The enzyme was purified 190-fold to near homogeneity. The purified enzyme appeared as a single band on 12.5% SDS-PAGE analysis with a molecular weight of 40,000 daltons. (c) 1995 John WileySons, Inc.

Calcineurin is an important signal transduction mediator in T cells, neurons, the heart, and kidneys. Recent evidence points to unique actions of the two main isoforms of the catalytic subunit. Although the β isoform is required for T-cell development, α is important in the brain and kidney. In addition, mice lacking α but not β suffer from failure to thrive and early mortality. The purpose of this study was to identify the cause of postnatal death of calcineurin α null (CnAα −/− ) mice and to determine the mechanism of α activity that contributes to the phenotype. CnAα −/− mice and wild-type littermate controls were fed a modified diet and then salivary gland function and histology were examined. In vitro studies were performed to identify the mechanism of α action. Data show that calcineurin is required for normal submandibular gland function and secretion of digestive enzymes. Loss of α does not impair nuclear factor of activated T-cell activity or expression but results in impaired protein trafficking downstream of the inositol trisphosphate receptor. These findings show a novel function of calcineurin in digestion and protein trafficking. Significantly, these data also provide a mechanism to rescue to adulthood a valuable animal model of calcineurin inhibitor-mediated neuronal and renal toxicities.

PGC-1alpha is a transcriptional coactivator that controls energy homeostasis through regulation of glucose and oxidative metabolism. Both PGC-1alpha expression and oxidative capacity are decreased in skeletal muscle of patients and animals undergoing atrophy, suggesting that PGC-1alpha participates in the regulation of muscle mass. PGC-1alpha gene expression is controlled by calcium- and cAMP-sensitive pathways. However, the mechanism regulating PGC-1alpha in skeletal muscle during atrophy remains unclear. Therefore, we examined the mechanism responsible for decreased PGC-1alpha expression using a rodent streptozotocin (STZ) model of chronic diabetes and atrophy. After 21days, the levels of PGC-1alpha protein and mRNA were decreased. We examined the activation state of CREB, a potent activator of PGC-1alpha transcription, and found that phospho-CREB was paradoxically high in muscle of STZ-rats, suggesting that the cAMP pathway was not involved in PGC-1alpha regulation. In contrast, expression of calcineurin (Cn), a calcium-dependent phosphatase, was suppressed in the same muscles. PGC-1alpha expression is regulated by two Cn substrates, MEF2 and NFATc. Therefore, we examined MEF2 and NFATc activity in muscles from STZ-rats. Target genes MRF4 and MCIP1.4 mRNAs were both significantly reduced, consistent with reduced Cn signaling. Moreover, levels of MRF4, MCIP1.4, and PGC-1alpha were also decreased in muscles of CnAalpha-/- and CnAbeta-/- mice without diabetes indicating that decreased Cn signaling, rather than changes in other calcium- or cAMP-sensitive pathways, were responsible for decreased PGC-1alpha expression. These findings demonstrate that Cn activity is a major determinant of PGC-1alpha expression in skeletal muscle during diabetes and possibly other conditions associated with loss of muscle mass.

This paper describes the relation between the Secondary Dendrite Arm Spacing (SDAS), Area of Mushy Zone with the Continuous Casting variables in low carbon steels during the solidification process in the mold zone. A Finite Element analysis of the heat flow equation, coupled with the solute distribution model and the dendrite growth model, enables the determination of the Secondary Dendrite Arm Spacing (SDAS). The CONBCAST.FOR program is developed in this work to analyze effects of process variables on the Secondary Dendrite Arm Spacing (SDAS), Area of Mushy Zone and Volume of the Bleed. Effort is also made to find the optimum casting parameters. A new concept is introduced in this work to analyze the relation between the Area of Mushy Zone and Secondary Dendrite Arm Spacing (SDAS) with the Volume of the Bleed. Quantitative work is performed by collecting the square shaped billets at two different process conditions and determined the Secondary Dendrite Arm Spacing and Volume of Bleed to analyze the relationship between SDAS, Area of Mushy zone and Volume of Bleed.Copyright © 2009 by ASME

Inflation-extension experiments were carried out on segments of the descending thoracic aortas from 4 normotensive and 4 hypertensive dogs rendered hypertensive using either unilateral or bilateral renal artery constriction. Intravascular pressures up to 200 mm Hg and axial forces up to 200 g were used. The external diameter of the segment and the distance between two longitudinally spaced gage marks were recorded photographically at each pressure-force level combination. Dimensions in the undeformed configuration were measured at the end of the inflation-extension experiment. Data were analyzed for changes in geometry and force-deformation response. Results indicate that: 1. Under sustained hypertension the wall thickness in the undeformed configuration increases with a concurrent reduction in the in-situ longitudinal extension ratio. 2. This dual tissue response accomplishes substantial reductions in the circumferential and longitudinal stresses from the levels that would be reached at equivlaent pressures in the absence of these geometric changes. 3. At comparable intravascular pressures the extensibility in the circumferential direction is slightly greater for the hypertensive aortas as compared to normals. However, the stress-extension ratio relationship in the circumferential direction is similar in the two groups. 4. The stress-extension ratio relationship in the longitudinal direction indicates that the hypertensive aorta is stiffer than its normotensive counterpart.

Summary The limitations of the conventional methods for diagnosing tuberculosis have spurred multi-faceted research activities in this field throughout the world. Chromatographic methods appear promising but may not be widely available in the developing countries. Immuno-diagnostic methods using combinations (“cocktails”) of antigens have high sensitivity and specificity and can easily be applied in the peripheral laboratories and in the field settings. Though expensive, molecular methods for diagnosis of tuberculosis have advantages of speed, sensitivity, and specificity. Adequate training of the eligible personnels in molecular methods and prevention of laboratory-dependent contamination may help reduce false positive results. Although, there are no clear guidelines, so far on how to make out the best from the gene amplification methods, yet their use may be encouraged with adequate quality controls, because of the inherent ingenuity and promises of these methods. Phage-based molecular methods provide rapid results in susceptibility tests for anti-tubercular drugs. In future, many sophisticated techniques are expected to hit the market for a rapid diagnosis of tuberculosis. In the developing countries, it is necessary to evaluate availability of suitable infrastructure and trained personnels before adopting modern diagnostic methods.

6-alpha-Hydroxy-7-deoxy-10-deacetylbaccatin-III is an intermediate that is potentially useful for synthesis of analogues of paclitaxel. Screening of microbial strains identified an enzyme activity in Nocardioides luteus SC 13912 (A.T.C.C. 55426) which converted 7-deoxy-10-deacetylbaccatin-III into 6-hydroxy-7-deoxy-10-deacetylbaccatin-III with a maximum yield of 44%.

Background Hyperhomocysteinemia has been implicated as an independent risk factor for atherosclerotic cardiovascular disease. Both cardiac transplantation and diabetes are associated with accelerated coronary artery disease. We therefore, designed this study to determine serum homocysteine levels in cardiac transplant recipients and patients with diabetes. We also sought to determine the relationship between hyperhomocysteinemia and renal insufficiency and its response to oral supplementation with folic acid.