Association of Serum Vitamin D level with Asthenozoospermic Male.

Although vitamin D deficiency is one of the most common health problems throughout the world, conflicting information exists on the potential association between serum vitamin D levels and semen quality. Currently available data identifies that vitamin D has a vital role in reproductive process as it affects sperm motility. This study was done with the rationality to evaluate the association between serum vitamin D levels with asthenozoospermic males. This cross-sectional analytic study was conducted on 314 men who attended the Department of Reproductive Endocrinology and Infertility, Bangabandhu Sheikh Mujib Medical University (BSMMU), Bangladesh July 2018 to June 2019. Considering the inclusion and exclusion criteria all participants were categorized into two groups; Group I included 157 asthenozoospermic male and Group II included 157 normozoospermic male according to World Health Organization 'strict' criteria 2010. Participants completed the questionnaires after they had agreed on a informed consent. Blood and semen samples were obtained for assessment and all data were adjusted for age, body mass index (BMI), total motility and progressive motility. Vitamin D levels were classified according to the Endocrine Society guideline. Statistical analyses were carried out by using the Statistical Package for Social Sciences version 22.0 for Windows (SPSS Inc., Chicago, Illinois, USA). The results showed that the mean vitamin D level was 16.63±5.54ng/ml in asthenozoospermic group and 19.83±5.33ng/ml in normozoospermic group. The mean vitamin D level was significantly (p<0.05) lower in asthenozoospermic group. It was noticed that 86.6% patients had vitamin D deficiency (≤20ng/ml) in asthenozoospermic group compared to 56.7% in the normozoospermic group. The study found that low vitamin D was associated with a fivefold increased risk of developing asthenozoospermia at 95% CI (2.74-8.99). Moreover, there was a positive significant correlation (r=0.285; p<0.001) between serum vitamin D level with total motility and progressive motility (r=0.232; p<0.001). Hence, the study suggests a significant association between asthenozoospermia and low vitamin D levels. However, clinical trials are warranted to further reinforce the findings.

Mycorrhization in trees: ecology, physiology, emerging technologies and beyond.

Mycorrhization has been an integral part of plants since colonization by the early land plants. Over decades, substantial research has highlighted its potential role in improving nutritional efficiency and growth, development and survival of crop plants. However, the focus of this review is trees. Evidence have been provided to explain ecological and physiological significance of mycorrhization in trees. Advances in recent technologies (e.g., metagenomics, artificial intelligence, machine learning, agricultural drones) may open new windows to apply this knowledge in promoting tree growth in forest ecosystems. Dual mycorrhization relationships in trees and even triple relationships among trees, mycorrhizal fungi and bacteria offer an interesting physiological system to understand how plants interact with other organisms for better survival. Besides, studies indicate additional roles of mycorrhization in learning, memorizing and communication between host trees through a common mycorrhizal network (CMN). Recent observations in trees suggest that mycorrhization may even promote tolerance to multiple abiotic (e.g., drought, salt, heavy metal stress) and biotic (e.g. fungi) stresses. Due to the extent of physiological reliance, local adaptation of trees is heavily impacted by the mycorrhizal community. This knowledge opens the possibility of a non-GMO avenue to promote tree growth and development. Indeed, mycorrhization could impact growth of trees in nurserys and subsequent survival of the inoculated trees in field conditions. Future studies might integrate hyperspectral imaging and drone technologies to identify tree communities that are deficient in nitrogen and spray mycorrhizal spore formulations on them.

Relation between Acute Ischemic Stroke and Left Ventricular Systolic Dysfunction.

Stroke is one of the leading causes of mortality and morbidity all over the world. The economic cost for stroke disability and post stroke rehabilitation is a growing concern. Ischemic stroke comprises 80.0 - 85.0% of total stroke cases caused by thrombotic or embolic occlusion of cerebral arteries. The source of embolism may be a larger artery or cardiac. Overt heart failure is an independent predictor of long term unfavorable functional outcome in stroke patients. However, there is little research whether the acute ischemic stroke risk is associated with mild to moderate degree of Left Ventricular Systolic Dysfunction (LVSD). This study was aimed to investigate the relation between LVSD and acute ischemic stroke (AIS) and to evaluate the relation between left ventricular systolic dysfunction and severity of neurological deficits after acute ischemic stroke. This case-control study was carried out in the Department of Neurology and Department of Medicine, Mymensingh Medical College and Hospital, Bangladesh from January 2019 to December 2020. One hundred twenty (120) patients of first ever AIS and 120 age and sex matched apparently healthy controls were enrolled in the study. Severity of stroke was measured by National Institute of Health Stroke Scale (NIHSS). Left ventricular (LV) systolic function was assessed by transthoracic 2-dimensional echocardiography. Mean±SD age was 58.23±9.34 years and 55.92±9.72 years respectively in cases and controls. Male to female ratio was 1.86:1. Left ventricular systolic dysfunction of any degree was more frequent in stroke patients (23.3%) than in controls (5.8%; p<0.001). The mean±SD of LVEF were 59.21±9.68 and 63.54±6.84 among case and control groups respectively. Mild LV dysfunction was observed 16.7% in AIS patients and 5.8% in control group. Moderate LV dysfunction was found in 6.7% in AIS patients. Participants with mild LVSD had significantly higher odds of being in the cases compared to participants with no LVSD (OR: 3.48; 95% CI: 1.41-8.59). Similarly, participants with moderate LVSD were 9.74 times more likely to be in ischemic stroke group compared to participants with no LVSD (OR: 9.74; 95% CI:1.19-19.29). LVSD was associated with AIS even after adjusting for other stroke risk factors (OR: 2.7462; 95CI for OR: 1.0204, 7.3906; p=0.0435). The NIHSS was significantly negatively correlated with LVEF (r=-0.443; p<0.001). The study may conclude that Left ventricular systolic dysfunction of mild to moderate degree, is associated with acute ischemic stroke. AIS patients with higher neurological deficit also had lower LVEF.

Relation of Neutrophil-Lymphocyte Ratio with Clinical Severity in Patients with Guillain-Barre Syndrome.

Guillain-Barre syndrome (GBS) is an acute autoimmune polyradiculoneuropathy. Neutrophil-lymphocyte ratio (NLR) is a novel prognostic and inflammatory marker in patients with neurological diseases. The study was designed to observe the relation between neutrophil lymphocyte ratio and the clinical severity in patients with Guillain-Barre Syndrome (GBS). This cross-sectional descriptive study was conducted at the department of Neurology and Medicine, Mymensingh Medical College and Hospital from April 2019 to September 2020. Total 58 patients with GBS were enrolled according to the inclusion and exclusion criteria within 7 days of development of symptoms. The clinical diagnosis of GBS was done according to Ausbury and Cornblath's diagnostic criteria; moreover clinical severity was done according to Hughes and Rees scale, Medical Research Council (MRC) grade, cranial nerve involvement and autonomic involvement. After measuring complete blood count, NLR was calculated by dividing neutrophil count to lymphocyte count. Data analysis was done on SPSS 23.0. Mean age of the GBS patients was 36.21±11.55 years. Among 58 respondents 70.69% (41) were male and 29.31% (17) were female. Most of the patients had a GBS severity score of 4(62.07%) followed by 3(27.59%) and 5(10.34%). The mean NLR of the respondents was 3.22±2.25. Acute motor axonal neuropathy (AMAN) was in 48.28% respondents and their mean NLR was 3.89±0.31, 31.03% had AIDP and mean NLR was 3.28±0.46 and 20.69% had Acute motor sensory axonal neuropathy (AMSAN) and mean NLR was 4.5±0.52. The mean NLR of the MRC grade 0, 1, 2, 3 patients was 6.61±1.78, 3.39±0.92, 2.71±1.39 and 1.45±0.40 respectively. Hughes score had a positive correlation (r=0.5333 and p<0.01) and MRC grade had a negative correlation (r= - 0.76805 and p<0.01) with NLR. Severity of GBS was significantly associated with raised NLR. Increase Hughes and Rees scale and decrease MRC grade also related to increased NLR.

Occurrence of canine parvovirus type 2c in diarrhoeic faeces of dogs in Kolkata, India.

Canine parvovirus-2(CPV-2) causes a highly contagious disease of dogs characterised by acute hemorrhagic gastroenteritis, lethargy, vomiting, fever and usually bloody or mucoid diarrhoea. In the present study, 41 faecal samples collected from dogs exhibiting the signs of fever, vomition, bloody or mucoid diarrhoea in Kolkata, India were screened by haemagglutination test and PCR for detection of capsid protein coding VP2 gene. The viral genotype was detected by multiplex PCR and analysis of partial VP2 gene nucleotide sequences of selected PCR products with bioinformatics tool. Thirteen (31.71%) samples were found positive with HA titre ≥ 32 whereas 28 (68.29%) samples were positive by PCR of VP2 gene indicating higher sensitivity of PCR. Highest occurrence of CPV-2 was observed in the age group of 1-6 months (80.65%) and non-descript breeds with no history of vaccination (85%). Three samples were antigenic type CPV-2a, rest were CPV-2b/CPV 2c. Six CPV sequences were found to be highly similar to published CPV 2c sequences in BLAST analysis revealing a maximum identity of 99-100% with other CPV-2c strains and clustered together with CPV-2c strains of India and other countries in phylogenetic analysis. The present study highlights the need for continuous monitoring of samples to detect gradual changes in circulating CPV-2 genotypes in India.

Association between Serum Vitamin D and Metabolic Syndrome in Women with Polycystic Ovary Syndrome.

Vitamin D deficiency (VDD) is common in women with and without polycystic ovary syndrome (PCOS) and may be associated with metabolic and endocrine disorders in PCOS. Aim of the study was to evaluate the association between serum vitamin D level and metabolic syndrome (MS) in PCOS. This cross-sectional study was conducted in PCOS clinic of infertility outpatient department of the department of Reproductive Endocrinology and Infertility at Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from July 2018 to June 2019. A total of 170 diagnosed patients with PCOS of aging 20 to 40 years were included in this study. Out of which 85 patients with PCOS MS and rest 85 patients without PCOS MS were considered as Group I and Group II respectively. Serum vitamin D was measured using a chemiluminescent microparticle immunoassay (CMIA). Almost two third (72.9%) populations had vitamin D moderate deficiency (10-20ng/ml) in Group I and 46(54.1%) in Group II. The mean vitamin D was 13.97±4.53ng/ml in Group I 16.02±6.62ng/ml in Group II. There were significant negative correlation found between vitamin D level with weight (r = -0.169; p=0.028), waist circumference (r = -0.208; p=0.007), Waist/Hip ratio (r = -0.205; p=0.007) and diastolic blood pressure (r = -0.187; p=0.014). In binary logistic regression analysis diastolic blood pressure (OR=1.04, 95% CI 0.89-1.29), fasting Glucose (OR=1.09, 95% CI 0.89 - 1.42), HDL (OR=1.11, 95% CI 1.05 - 1.17) and Vitamin D (OR=1.37, 95% CI 1.01 - 1.82) were significant risk factors of metabolic syndrome of the study patients. Vitamin D level was lower in PCOS subjects having MS and higher frequency of body weight, vitamin D deficiency was observed in them compared to the non-MS group. Waist: hip ratio, waist circumference and diastolic blood pressure were the negatively correlated with of vitamin D in the study subjects.

Mortality Attributable to Ambient Air Pollution: A Review of Global Estimates.

Since the publication of the first epidemiological study to establish the connection between long-term exposure to atmospheric pollution and effects on human health, major efforts have been dedicated to estimate the attributable mortality burden, especially in the context of the Global Burden of Disease (GBD). In this work, we review the estimates of excess mortality attributable to outdoor air pollution at the global scale, by comparing studies available in the literature. We find large differences between the estimates, which are related to the exposure response functions as well as the number of health outcomes included in the calculations, aspects where further improvements are necessary. Furthermore, we show that despite the considerable advancements in our understanding of health impacts of air pollution and the consequent improvement in the accuracy of the global estimates, their precision has not increased in the last decades. We offer recommendations for future measurements and research directions, which will help to improve our understanding and quantification of air pollution-health relationships.

Clinical efficacy of the virtual fracture clinic: analysis of 17,269 referrals by type of injury.

INTRODUCTION: Numerous studies have deemed the virtual fracture clinic (VFC) model to be both cost and clinically effective. However, very few of these studies have analysed the type of injuries seen in the VFC. The objectives of this study were to assess the clinical effectiveness of the VFC and analyse the types of injuries that lead to patients re-presenting in the face-to-face fracture clinic after being discharged virtually. METHODS: This is a retrospective study analysing 17,269 patients referred to the VFC between September 2017 and February 2020. Data regarding the type of presenting injury were collected to understand which injuries required further management after being discharged virtually. Patient clinic letters provided data regarding the purpose and outcomes of VFC referrals as well as face-to-face appointments. Theatre lists were cross-referenced to extract data regarding surgical management. RESULTS: In total, 57.37% (9,908) patients were discharged virtually. Of these patients, 92.52% were discharged successfully and 7.48% re-presented to the fracture clinic: 98.11% were managed conservatively and 1.88% required surgery. The highest number of failed discharges were for distal radius fractures (109, 14.69%). Face-to-face follow-up in fracture clinic was requested for 37.06% (6,400) of patients; 4.98% of them required surgical intervention. Some 5.56% (961) of referrals were removed from our analysis: 807 were inappropriate referrals and 154 were deemed suitable for multidisciplinary team discussion. The trust has saved an average of £702,205 annually since introduction of the VFC. CONCLUSIONS: The VFC model delivers as promised in terms of clinical efficacy and cost management. Injury types showing higher numbers of unsuccessful discharges could benefit from having modified management pathways.

PDGFRß promotes oncogenic progression via STAT3/STAT5 hyperactivation in anaplastic large cell lymphoma.

BACKGROUND: Anaplastic large cell lymphoma (ALCL) is an aggressive non-Hodgkin T cell lymphoma commonly driven by NPM-ALK. AP-1 transcription factors, cJUN and JUNb, act as downstream effectors of NPM-ALK and transcriptionally regulate PDGFRß. Blocking PDGFRß kinase activity with imatinib effectively reduces tumor burden and prolongs survival, although the downstream molecular mechanisms remain elusive. METHODS AND RESULTS: In a transgenic mouse model that mimics PDGFRß-driven human ALCL in vivo, we identify PDGFRß as a driver of aggressive tumor growth. Mechanistically, PDGFRß induces the pro-survival factor Bcl-xL and the growth-enhancing cytokine IL-10 via STAT5 activation. CRISPR/Cas9 deletion of both STAT5 gene products, STAT5A and STAT5B, results in the significant impairment of cell viability compared to deletion of STAT5A, STAT5B or STAT3 alone. Moreover, combined blockade of STAT3/5 activity with a selective SH2 domain inhibitor, AC-4-130, effectively obstructs tumor development in vivo. CONCLUSIONS: We therefore propose PDGFRß as a novel biomarker and introduce PDGFRß-STAT3/5 signaling as an important axis in aggressive ALCL. Furthermore, we suggest that inhibition of PDGFRß or STAT3/5 improve existing therapies for both previously untreated and relapsed/refractory ALK+ ALCL patients.

Punzi-loss:: a non-differentiable metric approximation for sensitivity optimisation in the search for new particles.

We present the novel implementation of a non-differentiable metric approximation and a corresponding loss-scheduling aimed at the search for new particles of unknown mass in high energy physics experiments. We call the loss-scheduling, based on the minimisation of a figure-of-merit related function typical of particle physics, a Punzi-loss function, and the neural network that utilises this loss function a Punzi-net. We show that the Punzi-net outperforms standard multivariate analysis techniques and generalises well to mass hypotheses for which it was not trained. This is achieved by training a single classifier that provides a coherent and optimal classification of all signal hypotheses over the whole search space. Our result constitutes a complementary approach to fully differentiable analyses in particle physics. We implemented this work using PyTorch and provide users full access to a public repository containing all the codes and a training example.

Fruit transcriptional profiling of the contrasting genotypes for shelf life reveals the key candidate genes and molecular pathways regulating post-harvest biology in cucumber.

Cucumber fruits are perishable in nature and become unfit for market within 2-3 days of harvesting. A natural variant, DC-48 with exceptionally high shelf life was developed and used to dissect the genetic architecture and molecular mechanism for extended shelf life through RNA-seq for first time. A total of 1364 DEGs were identified and cell wall degradation, chlorophyll and ethylene metabolism related genes played key role. Polygalacturunase (PG), Expansin (EXP) and xyloglucan were down regulated determining fruit firmness and retention of fresh green colour was mainly attributed to the low expression level of the chlorophyll catalytic enzymes (CCEs). Gene regulatory networks revealed the hub genes and cross-talk associated with wide variety of the biological processes. Large number of SSRs (21524), SNPs (545173) and InDels (126252) identified will be instrumental in cucumber improvement. A web genomic resource, CsExSLDb developed will provide a platform for future investigation on cucumber post-harvest biology.

Bilateral obstructive megaureter with right lower ureteric urothelial carcinoma: a very rare coalition.

Upper tract urothelial cancers account for 5% of all urothelial cancers. Among these, more than 90% of tumours are located in the renal pelvis, while ureteric tumours are extremely rare. The distal ureter is the most common location, although multifocal implants may also occur. Megaureter is a common cause of obstructive uropathy in neonates and children. Nevertheless, it may be unnoticed, if asymptomatic. Megaureter may be obstructing or refluxing. Long-standing urinary stasis and recurrent urinary tract infection in megaureter may cause chronic irritation of the ureteric mucosa leading to dysplasia and malignancy. We report a 55-year-old man diagnosed with bilateral obstructive megaureter with right lower ureteric urothelial cancer and review the current literature.

Versatility Of Square Flaps In Post-Burn Contractures Involving Upper Limb.

Post-burn contractures are a common problem and functionally limiting for upper limbs. Many different techniques have been described in the literature for their treatment. Z-plasty and release with SSG cover are the commonest procedures done for post-burn contractures. In this study we assess the use of the square flap technique in post-burn contractures of upper limb. Eleven patients with a total number of twelve upper limb contractures (mild to moderate) involving axilla, elbow and finger underwent release by standard square flap technique. All cases were followed up for at least 6 months and analyzed for range of motion and aesthetic outcome. Patient and surgeon satisfaction was recorded. All operated cases achieved a satisfactory range of movement post-operatively without any recurrence. The number of patients who were satisfied with the surgery were 7 out of 11, and 4 patients were somewhat satisfied with the results obtained. In contrast, the surgeons were satisfied in all cases. Square flap is shown to be an easy and reliable flap for mild to moderate contractures of the anterior or posterior axillary folds, elbow contractures and finger contractures with low recurrence rate.

Les rétractions séquellaires de brûlure sont fréquentes et engendrent des limitations fonctionnelles au membre supérieur. Beaucoup de techniques différentes ont été décrites dans la littérature pour leur traitement : plastie en Z, libération et couverture par greffe de peau, sont les techniques les plus couramment utilisées. Dans cette étude, nous évaluons l'usage du double lambeau rhomboïde ou lambeau carré de Hyakusoku dans les rétractions séquellaires de brûlure du membre supérieur. 11 patients représentant 12 rétractions au membre supérieur (minimes à modérées) intéressant la région axillaire, le coude, et les doigts ont bénéficié d'une libération de la rétraction par la technique du double lambeau rhomboïde ou lambeau carré de Hyakusoku. Tous les patients ont été suivis pendant au moins six mois post-opératoire. L'analyse porte à la fois sur les mobilités et le résultat esthétique. La satisfaction du patient et du chirurgien a été notée. Toutes les interventions ont permis une nette amélioration de la fonction, sans récidive. 7 patients sur 11 ont été satisfaits par le résultat du geste chirurgical et quatre patients ont été assez satisfaits. En revanche, les chirurgiens ont noté les résultats comme satisfaisants dans tous les cas. Le double lambeau rhomboïde ou lambeau carré de Hyakusoku semble donc être un lambeau simple et fiable dans le traitement des rétractions minimes à modérées des piliers axillaires antérieur ou postérieur, du coude ou des doigts avec un très faible taux de récidive.

Risk Association, Clinical Presentation and Management of Uterine Leiomyoma.

Uterine leiomyoma is a common gynecological problem throughout the world. As 50% hysterectomies in black and 40% in Australians are performed due to fibroid, in our country also it is assumed that about 40%-50% hysterectomies are done due to fibroid uterus. Thus leiomyoma constitute a major public health cost to the community in terms of outpatient attendance and hospital cost for surgery. This is a descriptive type of cross sectional study among 50 patients having leiomyoma of uterus in the department of Obs and Gynae, BSMMU hospital from March 2011 to August 2011. The objective of the study is to find out the risk associated with leiomyoma, to find out the common presenting features of uterine leiomyoma and to find out best options for management. Study results showed that 62% patients were in the age group 36-45 years, 48%patients were in para1-2 group, 32% patients used combined oral contraceptive pill for contraception. Sixty percent (60%) patients presented with progressive menorrhagia and palpable mass was found in 62% cases. About 46% patients had associated medical conditions like hypertension, diabetes, obesity. Total abdominal hysterectomy was done in 32% cases. TAH with unilateral or bilateral salpingoophorectomy was done in 40% cases. Myomectomy was done in 20% cases. There is a scope for large scale study about risk factors of uterine leiomyoma like obesity, diabetes mellitus, hypertension, use of hormonal contraceptive, racial differences, different treatment modalities etc. Treatment should be individualized. However in this connection a good referral system and good communication has got a tremendous contribution in the proper management of such problems.

Clinical Risk Index for Babies II Score as a Predictor of Neonatal Death among Preterm Low Birth Weight Babies.

Clinical risk index for babies II (CRIB II) score is simple, validated and widely used risk-adjustment instrument for predicting mortality among preterm low birth weight babies. To assess the efficacy of CRIB II score as a tool to predict the risk for neonatal death among the preterm and LBW babies admitted in NICU of BSMMU, a tertiary care hospital in Bangladesh. This prospective observational study was conducted in Department of Neonatology in BSMMU from September 2016 to August 2017. Inborn preterm neonates with gestational age ≤34 weeks admitted were enrolled in the study. CRIB-II score was calculated for each infant within 1 hour of birth from birth weight, gestational age, sex, admission temperature and base excess. The primary outcome measured in the study was neonatal death or survival up to 28 days. Total 112 patients were finally analyzed in this study. Mean CRIB II score was significantly higher in the non-survivor group compared to the survivor group (p-value <0.0001). Receiver operating characteristic (ROC) curve analysis for mortality prediction by CRIB II score, gestational age and birth weight showed AUC 0.87 (95% CI 0.76-0.97), 0.76 (95% CI 0.63-0.88) and 0.79 (95% CI 0.66-0.92) respectively. ROC curve analysis also revealed that the most suitable cut-off points for predicting mortality were 5 for CRIB II score, 32 weeks for gestational age and 1250 gram for birth weight. Using these most suitable cut-off points, CRIB II score had the highest sensitivity and specificity followed by birth weight and gestational age. In this study, CRIB II score was found to be an effective tool for predicting neonatal death among preterm LBW babies. It predicted outcome more accurately than birth weight or gestational age alone.

Obstetric Outcome of Multiple Pregnancy in a Medical College Hospital.

The Incidence of twin and high order multiple gestation has increased significantly over the past 15 years primarily because of the availability and increased use of ovulation inducing drugs and assisted reproductive technology. Both maternal and perinatal mortality and morbidity is higher in multiple pregnancy than in singleton pregnancy. Proposed study is designed to determine the incidence and obstetric outcome of multiple pregnancy which is a high-risk pregnancy in a tertiary referral hospital. The purpose of this study is to evaluate obstetric outcome of multiple pregnancy. This prospective observational study was carried out in the Department of Obs and Gynae, Mymensingh Medical College Hospital, Mymensingh from December 2011 to May 2012. Patients were selected by purposive way. In this study it was observed that incidence of twin pregnancy was 1.66% common age group 30-35 years (48%), common in multipara (74%), no pregnancy crossed EDD. Family history of twin was 32% and unknown etiology 58%. Most common maternal complication was preterm labour (60%). Commonest mode of delivery was LSCS (64%), incidence of perinatal mortality 15%, 36% baby had birth weight in between 2.1-2.5kg, all the baby requiring admission in neonatal unit. This prospective observational study revealed that perinatal mortality was high in our center, there was no maternal mortality.

Socio-demographic Profile of COVID-19 Patients and Difficulties Faced by Them while Testing RT-PCR: A Multi-center Cross-sectional Study.

This study was aimed to find out the socio-demographic profiles as well as difficulties of Reverse transcription polymerase chain reaction (RT-PCR) testing faced by Coronavirus Disease 2019 (COVID-19) patients. This cross-sectional study was conducted from 10th August to 7th September 2020. Data were collected by telephone interview using a pre-designed questionnaire after taking verbal consent from the participants. Out of 281 COVID-19 patients, 279 were diagnosed by RT-PCR; 10.3% were asymptomatic. Off them 67.6%were males, 24% were hospital staffs. Majority (66.2%) were from the non-city corporation area. History of recent contact with known COVID-19 patients was present in 56.9% cases. Fever (78.3%) and cough (58%) were the most common symptoms. One third of the patients faced difficulty to test RT-PCR for COVID-19. Sixteen percent patients complained of difficulty of getting serial for testing, the maximum waiting time was one week before giving samples. Thirty patients (10.8%) had to wait longer than usual time after reaching the center before giving sample. Hospital staffs were unable to co-operate in 2.5% of the patients while difficulty of managing transport to the hospital for suspected COVID-19 patient was an issue in 2.2% of the patients. Though testing was more difficult in city corporation areas (p=0.028), delay of getting test result was less (p<0.001). Maximum delay of getting test result was 10 days. Finding out the difficulties of COVID-19 testing will help to point out the issues behind these and will help to take necessary steps to tackle this matter. Testing rate can be increased to contain this highly contagious virus in this densely populated country.

Antibiofilm and anti-quorum sensing activities of eugenol and linalool from Ocimum tenuiflorum against Pseudomonas aeruginosa biofilm.

AIMS: The aim of this study is to determine the ability of two bioactive compounds, namely, eugenol and linalool, purified from leaves of Ocimum tenuiflorum for eradication of biofilm produced by Pseudomonas aeruginosa. METHODS AND RESULTS: The phytoextract of O. tenuiflorum (KT), a common ethno-botanical plant of India, was purified through high-performance liquid chromatography and was analysed using ultraviolet (UV) spectroscopy and gas chromatography-mass spectrometry (GC-MS). Eugenol and linalool were found to be the most active amongst all phytocompounds present in phytoextract and showed a significant reduction in the viability of sessile cells of P. aeruginosa and the minimum revival after withdrawal of phyto-challenge. They could bring about notable reduction in the protein and carbohydrate content of exopolysaccharide of biofilm. Eugenol and linalool could affect the synthesis of quorum sensing (QS) proteins like LasA and LasB as well as virulence factors such as pyocyanin, and rhamnolipids, which seriously hamper the formation of biofilm. The biofilm framework was extremely affected by the phytocompounds through the reduction of protein and carbohydrate content of extracellular polymeric substance (EPS). Another interesting found out was that they brought about maximum inhibition to the genomic DNA and RNA content. The studies were supported by in silico interaction between eugenol and linalool with the QS proteins. The antibiofilm efficacies of eugenol, linalool and phytoextract (KT) were further confirmed by microscopic studies with scanning electron microscopy (SEM), atomic force microscopy and fluorescence confocal microscopy microscopic studies. CONCLUSIONS: The phytocompounds are proved to be more effective than conventional antibiotics in inhibiting the biofilm forming sessile cells and can be used as a replacement for antibiotic. SIGNIFICANCE AND IMPACT OF THE STUDY: Pure eugenol extracted from common basil leaves can be used as a safe substitute for common antibiotic for treatment of chronic infections caused by P. aeruginosa. It will be cost effective, devoid of notable side effects and will not generate antibiotic resistance in host body.