BACKGROUND: A high prevalence of antimicrobial resistance among urinary isolates in the Garhwal region of Uttaranchal. AIMS: To identify the most appropriate antibiotic for empirical treatment of community-acquired acute cystitis on the basis of local antimicrobial sensitivity profile. SETTINGS AND DESIGN: A prospective clinico microbiological study including all clinically diagnosed patients with community acquired acute cystitis attending a tertiary care teaching hospital over a period of three years. METHODS AND MATERIAL: Clean-catch midstream urine specimens, from 524 non pregnant women with community-acquired acute cystitis, were subjected to semiquantitative culture and antibiotic susceptibility by the Kirby-Bauer disc diffusion method. A survey was also conducted on 30 randomly selected local practitioners, to know the prevalent prescribing habits in this condition. STATISTICAL ANALYSIS: The difference between the susceptibility rates of E. coli isolates to Nitrofurantoin and the other commonly prescribed antibiotics was analysed by applying the z test for proportion. RESULTS: 354 (67.5%) specimens yielded significant growth of E. coli. >35% of the urinary E.coli isolates were resistant to the fluoroquinolones, which were found to be the most commonly used empirical antibiotics in acute cystitis. Resistance was minimum against Nitrofurantoin (9.3%, 33) and Amikacin (11.0%,39). >80% of the fluoroquinolone-resistant strains were found to be sensitive to Nitrofurantoin. CONCLUSION: The best in vitro susceptibility profile in our study has been shown by Nitrofurantoin and a significantly high proportion of the urinary E. coli isolates have already developed resistance to the currently prescribed empirical antibiotics, viz. the fluoroquinolones. In view of these in vitro susceptibility patterns, a transition in empirical therapy appears imminent.

In this work, we report a facile synthesis of graphene oxide–gold (GO–Au) nanocomposites by electrodeposition. The fabricated electrochemical immunosensors are utilized for the dual detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen and SARS-CoV-2 antibody. The GO–Au nanocomposites has been characterized by UV–vis spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) for its biosensing properties. The linear detection range of the SARS-CoV-2 antigen immunosensor is 10.0 ag mL –1 to 50.0 ng mL –1 , whereas that for the antibody immunosensor ranges from 1.0 fg mL –1 to 1.0 ng mL –1 . The calculated limit of detection (LOD) of the SARS-CoV-2 antigen immunosensor is 3.99 ag mL –1 , and that for SARS-CoV-2 antibody immunosensor is 1.0 fg mL –1 with high sensitivity. The validation of the immunosensor has also been carried out on patient serum and patient swab samples from COVID-19 patients. The results suggest successful utilization of the immunosensors with a very low detection limit enabling its use in clinical samples. Further work is needed for the standardization of the results and translation in screen-printed electrodes for use in portable commercial applications.

The number of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) cases is increasing in India. This study looks upon the geographic distribution of the virus clades and variants circulating in different parts of India between January and August 2020. The NPS/OPS from representative positive cases from different states and union territories in India were collected every month through the VRDLs in the country and analyzed using next-generation sequencing. Epidemiological analysis of the 689 SARS-CoV-2 clinical samples revealed GH and GR to be the predominant clades circulating in different states in India. The northern part of India largely reported the ‘GH’ clade, whereas the southern part reported the ‘GR’, with a few exceptions. These sequences also revealed the presence of single independent mutations—E484Q and N440K—from Maharashtra (first observed in March 2020) and Southern Indian States (first observed in May 2020), respectively. Furthermore, this study indicates that the SARS-CoV-2 variant (VOC, VUI, variant of high consequence and double mutant) was not observed during the early phase of virus transmission (January–August). This increased number of variations observed within a short timeframe across the globe suggests virus evolution, which can be a step towards enhanced host adaptation.

Research on tuberculosis and leprosy was revolutionized by the development of a plasmid transformation system in the fast-growing surrogate, Mycobacterium smegmatis. This transformation system was made possible by the successful isolation of a M. smegmatis mutant strain mc 2 155, whose efficient plasmid transformation (ept) phenotype supported the replication of Mycobacterium fortuitum pAL5000 plasmids. In this report, we identified the EptC gene, the loss of which confers the ept phenotype. EptC shares significant amino acid sequence homology and domain structure with the MukB protein of Escherichia coli, a structural maintenance of chromosomes (SMC) protein. Surprisingly, M. smegmatis has three paralogs of SMC proteins: EptC and MSMEG_0370 both share homology with Gramnegative bacterial MukB; and MSMEG_2423 shares homology with Gram-positive bacterial SMCs, including the single SMC protein predicted for Mycobacterium tuberculosis and Mycobacterium leprae. Purified EptC was shown to bind ssDNA and stabilize negative supercoils in plasmid DNA. Moreover, an EptC-mCherry fusion protein was constructed and shown to bind to DNA in live mycobacteria, and to prevent segregation of plasmid DNA to daughter cells. To our knowledge, this is the first report of impaired plasmid maintenance caused by a SMC homolog, which has been canonically known to assist the segregation of genetic materials.plasmid segregation | electroporation | DNA topology | partition errors | cell division

From March to June 2021, India experienced a deadly second wave of COVID-19, with an increased number of post-vaccination breakthrough infections reported across the country. To understand the possible reason for these breakthroughs, we collected 677 clinical samples (throat swab/nasal swabs) of individuals from 17 states/Union Territories of the country who had received two doses (n = 592) and one dose (n = 85) of vaccines and tested positive for COVID-19. These cases were telephonically interviewed and clinical data were analyzed. A total of 511 SARS-CoV-2 genomes were recovered with genome coverage of higher than 98% from both groups. Analysis of both groups determined that 86.69% (n = 443) of them belonged to the Delta variant, along with Alpha, Kappa, Delta AY.1, and Delta AY.2. The Delta variant clustered into four distinct sub-lineages. Sub-lineage I had mutations in ORF1ab A1306S, P2046L, P2287S, V2930L, T3255I, T3446A, G5063S, P5401L, and A6319V, and in N G215C; Sub-lineage II had mutations in ORF1ab P309L, A3209V, V3718A, G5063S, P5401L, and ORF7a L116F; Sub-lineage III had mutations in ORF1ab A3209V, V3718A, T3750I, G5063S, and P5401L and in spike A222V; Sub-lineage IV had mutations in ORF1ab P309L, D2980N, and F3138S and spike K77T. This study indicates that majority of the breakthrough COVID-19 clinical cases were infected with the Delta variant, and only 9.8% cases required hospitalization, while fatality was observed in only 0.4% cases. This clearly suggests that the vaccination does provide reduction in hospital admission and mortality.

MicroRNAs (miRNAs) are small noncoding RNAs that are crucial posttranscriptional regulators for host mRNAs. Recent studies indicate that miRNAs may modulate host response during RNA virus infection. However, the role of miRNAs in immune response against H5N1 infection is not clearly understood. In this study, we showed that expression of cellular miRNA miR-324-5p was downregulated in A549 cells in response to infection with RNA viruses H5N1, A/PR8/H1N1, and Newcastle disease virus (NDV) and transfection with poly(I·C). We found that miR-324-5p inhibited H5N1 replication by targeting the PB1 viral RNA of H5N1 in host cells. In addition, transcriptome analysis revealed that miR-324-5p enhanced the expression of type I interferon, type III interferon, and interferon-inducible genes (ISGs) by targeting CUEDC2, the negative regulator of the JAK1-STAT3 pathway. Together, these findings highlight that the miR-324-5p plays a crucial role in host defense against H5N1 by targeting viral PB1 and host CUEDC2 to inhibit H5N1 replication. Highly pathogenic influenza A virus (HPAIV) continues to pose a pandemic threat globally. From 2003 to 2017, H5N1 HPAIV caused 453 human deaths, giving it a high mortality rate (52.74%). This work shows that miR-324-5p suppresses H5N1 HPAIV replication by directly targeting the viral genome (thereby inhibiting viral gene expression) and cellular CUEDC2 gene, the negative regulator of the interferon pathway (thereby enhancing antiviral genes). Our study enhances the knowledge of the role of microRNAs in the cellular response to viral infection. Also, the study provides help in understanding how the host cells utilize small RNAs in controlling the viral burden.

Background: In view of the role of TLR2 activation in host defense against mycobacteria, the present study was conducted to examine whether TLR2 polymorphisms could account for the increased prevalence of tuberculosis in Indian patients. Detection of such polymorphisms would help in assessing the risk of developing active tuberculosis among contacts or HIV positive patients and in identifying candidates for chemoprophylaxis.

COVID-19 belongs to a typical class of viruses that predominantly affects the human respiratory system, thereby proving to be fatal to many. The virus, along with other air pollutant particulates poses a severe threat to the human respiratory organs. Since the most common transmission mode is respiratory fomites and aerosol particulates, it is necessary to prevent their ingression through a mask. The primary use of masks is to prevent aerial particulates. This paper reveals the development of masks with air filters coated with functionalized graphene (fG) mounted on a 3D-printed facial mask replica. The fused deposition modeling (FDM) process is used for fabricating the facial mask replica. fG associated with nanosheets has an additional adsorbing capacity with a high surface area to volume ratio. fG coat is used over a polypropylene (PP) cloth through a dip coating method to enhance the antiviral and antimicrobial properties. The quality of fG is investigated through Raman spectroscopy and other characterization techniques such as SEM, XRD, and FTIR were used for visual interpretation of distributions of fG on a polypropylene (PP) fabric. Fabricated fG coated MB filters show 98.2 % of bacterial filtration efficiency with 1.10 mbar of breathing resistance. The efficacy of the fG coated filter is tested against SARS-CoV-2 viral particles, which shows a complete arrest of viral transmission at the fG coated layer.