Molecular Epidemiology and Characterization of Multidrug-Resistant MRSA ST398 and ST239 in Himachal Pradesh, India.

Aim: Methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of severe and difficult-to-treat infections in humans and animals. We aimed to identify the predominant lineages of methicillin-resistant S. aureus in Himachal Pradesh, India, to understand the genomic epidemiology along with the genotypic and phenotypic characteristics. Methods: We isolated 250 S. aureus from two district hospitals in Himachal Pradesh, India. Methicillin-Resistant S. aureus (MRSA) isolates were subjected to MLST, SCCmec typing, and resistance as well as virulence determinants were determined by PCR and sequencing. Bio-typing was also performed for source tracking. Results: A 17.6% (44/250) of isolates were classified as MRSA by both the MRSA detection kit and disc diffusion methods. Antimicrobial Susceptibility Testing of MRSA isolates (n = 44) showed high resistance to oxacillin (77.27%), erythromycin (77.27%), tetracycline (75%), cefoxitin (65.9%), and gentamicin (61.36%), while low resistance was observed for teicoplanin (36.36%), vancomycin and levofloxacin (31.81%) and fusidic acid (18.18%). All isolates were sensitive to linezolid, quinupristin-dulfopristin, dalbavancin, and cefazoline. The SCCmec-II was observed in 20.45% of isolates, SCCmec-I in 11.36%, SCCmec-III in 9%, SCCmec-IV in 40.9% and SCCmec-V in 18.18%. The mecA gene was present in all isolates (n = 44) and 50% also had the vanA gene. 35% of isolates had the lukS-PV/lukf-PV toxin gene and 11.36% had the co-existence of mecA, vanA, and lukS-PV/lukf-PV. The major strain was ST398 (39%) followed by ST239 (27%), ST217 (16%), ST121 (11%), and ST338 (7%). The MRSA isolates produced staphylokinase and ß-hemolysis but were negative for bovine plasma coagulation tests. In Conclusion: The predominant MRSA clones in Himachal Pradesh, India, were hospital-associated multi-drug resistant-MRSA ST239 with PVL and community-associated MRSA ST398.

Biology and molecular interactions of Parastagonospora nodorum blotch of wheat.

MAIN CONCLUSION: Parastagonospora nodorum is one of the important necrotrophic pathogens of wheat which causes severe economical loss to crop yield. So far, a number of effectors of Parastagonospora nodorum origin and their target interacting genes on the host plant have been characterized. Since targeting effector-sensitive gene carefully can be helpful in breeding for resistance. Therefore, constant efforts are required to further characterize the effectors, their interacting genes, and underlying biochemical pathways. Furthermore, to develop effective counter-strategies against emerging diseases, continuous efforts are required to determine the qualitative resistance that demands to screen of diverse genotypes for host resistance. Stagonospora nodorum blotch also refers to as Stagonospora glume blotch and leaf is caused by Parastagonospora nodorum. The pathogen deploys necrotrophic effectors for the establishment and development on wheat plants. The necrotrophic effectors and their interaction with host receptors lead to the establishment of infection on leaves and extensive lesions formation which either results in host cell death or suppression/activation of host defence mechanisms. The wheat Stagonospora nodorum interaction involves a set of nine host gene-necrotrophic effector interactions. Out of these, Snn1-SnTox1, Tsn1-SnToxA and Snn-SnTox3 are one of the most studied interaction, due to its role in the suppression of reactive oxygen species production, regulating the cytokinin content through ethylene-dependent wayduring initial infection stage. Further, although the molecular basis is not fully unveiled, these effectors regulate the redox state and influence the ethylene biosynthesis in infected wheat plants. Here, we have discussed the biology of the wheat pathogen Parastagonospora nodorum, role of its necrotrophic effectors and their interacting sensitivity genes on the redox state, how they hijack the resistance mechanisms, hormonal regulated immunity and other signalling pathways in susceptible wheat plants. The information generated from effectors and their corresponding sensitivity genes and other biological processes could be utilized effectively for disease management strategies.

CRISPR-Cas-Led Revolution in Diagnosis and Management of Emerging Plant Viruses: New Avenues Toward Food and Nutritional Security.

Plant viruses pose a serious threat to agricultural production systems worldwide. The world's population is expected to reach the 10-billion mark by 2057. Under the scenario of declining cultivable land and challenges posed by rapidly emerging and re-emerging plant pathogens, conventional strategies could not accomplish the target of keeping pace with increasing global food demand. Gene-editing techniques have recently come up as promising options to enable precise changes in genomes with greater efficiency to achieve the target of higher crop productivity. Of genome engineering tools, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) proteins have gained much popularity, owing to their simplicity, reproducibility, and applicability in a wide range of species. Also, the application of different Cas proteins, such as Cas12a, Cas13a, and Cas9 nucleases, has enabled the development of more robust strategies for the engineering of antiviral mechanisms in many plant species. Recent studies have revealed the use of various CRISPR-Cas systems to either directly target a viral gene or modify a host genome to develop viral resistance in plants. This review provides a comprehensive record of the use of the CRISPR-Cas system in the development of antiviral resistance in plants and discusses its applications in the overall enhancement of productivity and nutritional landscape of cultivated plant species. Furthermore, the utility of this technique for the detection of various plant viruses could enable affordable and precise in-field or on-site detection. The futuristic potential of CRISPR-Cas technologies and possible challenges with their use and application are highlighted. Finally, the future of CRISPR-Cas in sustainable management of viral diseases, and its practical utility and regulatory guidelines in different parts of the globe are discussed systematically.

Genetic analysis of maydis leaf blight resistance in subtropical maize (Zea mays L.) germplasm.

Knowledge on the genetics of maydis leaf blight (MLB) is crucial to breed the resistant maize cultivars to combat disease epidemics as a sustainable and cost-effective approach. The present investigation was framed to understand the genetics of MLB resistance in subtropical maize. Two contrasting genotypes CM119 (susceptible) and SC-7-2-1-2-6-1 (resistant) were used to generate six genetic populations, namely P1, P2, F1, F2, BC1P1 and BC1P2, and evaluated in three target environments for MLB resistance under artificial epiphytotic condition. The CM119 and SC-7-2-1-2-6-1 showed susceptible and resistant reactions with mean disease reaction of 3.89-3.98 and 1.88-2.00, respectively. The derived generations, namely F1, F2, BC1P1 and BC1P2 showed mean disease reaction of 2.15-2.28, 2.44-2.51, 2.19-2.24 and 2.22-2.28, respectively in the test locations. The segregating generations (F2: 0.35-0.37; BC1P1: 0.24-0.29 and BC1P2: 0.17-0.20) showed variation for MLB disease resistance over the parental and first filial generations (P1: 0.11-0.17; P2: 0.08-0.13 and F1: 0.12-0.14). The genetic analysis of MLB resistance revealed the nonallelic interactions of duplicate epistasis type across the test locations. Among the gene interactions, dominance x dominance [l] effect was predominant over additive x additive [i] and additive x dominance [j] effects. The segregation analysis and the prediction of the number of major loci revealed at least two major genes associated with MLB tolerance in subtropical maize. Our investigation paved the foundation for the improvement of subtropical maize germplasm of MLB resistance.

Differential Pulse Polarographic Investigations on Glyphosate and Glufosinate Herbicides in Relation to Their Environmental Analysis.

Differential pulse polarographic (DPP) investigations on the reaction of the amino function of glyphosate and glufosinate herbicides with carbon disulphide and copper(II) perchlorate forming copper(III) dithiocarbamate complexes were made in the presence of sodium perchlorate in acetonitrile at dropping mercury electrode (DME). The newly formed herbicide complexes exhibited analytically useful diffusion-controlled peaks at - 115 mV and - 110 mV versus saturated calomel electrode (SCE) with linear relationship between current and concentration. This observation formed the basis for the determination of glyphosate and glufosinate in the concentration ranges 0.34-8.45 µg mL-1 and 0.4-9.91 µg mL-1 respectively with correlation coefficient of 0.999. The method was applied to their determinations in soil, fortified food and spiked water samples to assess their environmental relevance. The recoveries of the herbicides were in the range 89.5%-98.3% with relative standard deviation (RSD) in the ranges 0.8%-1.8% thus showing good accuracy and precision of the method.

Clinical Characteristics and Outcome Trends of Adjuvant Anthracycline and Taxane Regimen for Early Stage Breast Cancer.

BACKGROUND: The anthracycline and taxane-based chemotherapy treatment regimen remains the gold standard for treatment of early stage breast cancer. However, studies examining the effectiveness and use of this treatment regimen in Indian context are limited. This study examined patients treated with anthracycline and taxane-based chemotherapy at a tertiary care cancer center in India. METHODS: Patients with confirmed early stage breast cancer who had undergone primary breast surgery followed by treatment with anthracycline and taxane-based chemotherapy between 2009 and 2015 were included in the study. Data on clinical characteristics and treatment details were collected from the patients' medical records. RESULTS: Two hundred sixty-four women were included in the analysis. The median age at presentation was 50 years. Among the 264 women, 40.5% were premenopausal, 1.2% were perimenopausal, and 58.3% were postmenopausal. The number of patients undergoing breast-conserving surgery (BCS) and modified radical mastectomy (MRM) were 35.2% and 64.7%, respectively. Patients with a tumor grade of 1, 2, and 3 were 7.2%, 53.1%, and 39.7%, respectively. Tumors were unifocal in 81.1% and multifocal in 18.2% of patients. Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER-2) positivity was detected in 58.3%, 54.2%, and 3.1% of patients, respectively and 38.6% of patients were triple negative. With a median follow-up of 36.2 months, the invasive disease-free survival rate was 90.9% and mean disease-free survival time was 65.4 ± 1.13 months. CONCLUSIONS: The results of this study confirm the clinical utility of anthracycline and taxane-based chemotherapy regimen as the adjuvant chemotherapy treatment of early stage breast cancer.

Comparative evaluation of the antibacterial and physical properties of conventional glass ionomer cement containing chlorhexidine and antibiotics.

OBJECTIVE: To evaluate the antimicrobial efficacy and compressive strength of conventional glass ionomer cement (GIC) containing chlorhexidine and antibiotics at varying concentrations. MATERIALS AND METHODS: Chlorhexidine diacetate and antibiotics (ciprofloxacin, metronidazole, and minocycline) were incorporated into GIC Fuji IX at 1.5% and 3% w/w ratio to form the experimental groups. The experimental GIC specimens were placed on brain heart infusion agar plates inoculated with Streptococcus mutans, and the area of inhibition was measured after 48 h. The 24-h compressive strength of the set specimens was evaluated using a Universal Testing Machine. RESULTS: The control group demonstrated no zone of inhibition. All experimental groups showed inhibition against S. mutans (P < 0.05), with larger zones of inhibition found in the higher concentration groups. Compressive strength at the end of 24 h decreased in the experimental groups as compared to the control group (P < 0.05), but no difference was found between the experimental groups (P > 0.05). CONCLUSION: The present study demonstrated that experimental GICs containing chlorhexidine diacetate and antibiotics were effective in inhibiting S. mutans, and incorporation of 1.5% ABX was optimal to give the appropriate antibacterial and physical properties.

Mediastinal germ cell tumour causing superior vena cava tumour thrombosis.

We report a rare case of a 35-year-old man who presented with a 1-week history of retrosternal chest pain of moderate intensity. A positron emission tomography CT (PET-CT) showed a large fluorodeoxy-glucose (FDG)-avid heterogeneously enhancing necrotic mass in the anterosuperior mediastinum with a focal FDG-avid thrombosis of the superior vena cava (SVC) suggestive of tumour thrombus and vascular invasion. α-Fetoprotein levels were raised (5690 IU/L). Image guided biopsy of the mediastinal mass was suggestive of non-seminomatous germ cell tumour (NSGCT). The patient received four cycles of BEP (bleomycin, etoposide and cisplatin) along with therapeutic anticoagulation with low-molecular-weight heparin. Follow-up whole body PET-CT revealed complete resolution of mediastinal mass and SVC tumour thrombosis. The documentation of FDG-PET-avid tumour thrombus resolving with chemotherapy supports the concept of circulating tumour cells being important not only in common solid tumours such as breast and colon cancer but also in relatively less common tumours such as NSGCT. The detection of circulating tumour cells could help deploy aggressive regimens upfront.

Whole cell pertussis vaccine: reducing toxicity.

Since introduction of the pertussis vaccine in 1940's the morbidity and mortality due to the infection has been markedly reduced all over the world. However the adverse effects of the inactivated whole cell pertussis vaccine like pain, swelling at the site of injection, fever, vomiting anorexia, persistent crying & drowsiness have been the cause of great concern, till date. Also the safety concerns over the use of thiomersal as an inactivating agent as well preservative have been raised in the recent past. Studies in many countries have been initiated to reduce or replace thiomersal & using other inactivating agents in the vaccines. Limited studies have been conducted in India. The present study has been undertaken as an attempt to reduce the quantity of thiomersal and modification in the procedure of production of the pertussis vaccine to reduce the toxicity, to produce better quality of whole cell pertussis vaccine. To achieve this, at the time of production of the whole cell pertussis vaccine, at Kasauli, as per the standard procedure recommended by WHO, three parallel batches of the pertussis culture were inactivated in fermenter before harvesting and thiomersal was used only one time for suspending the bacterial mass after harvesting. The resultant modified vaccine so prepared when tested showed that it was of better quality as compared to the one produced by standard procedure, when tested in mice.

Sigmoid vaginoplasty: long-term results.

OBJECTIVES: To evaluate the long-term results of sigmoid vaginoplasty for Mayer-Rokitansky-Kuster-Hauser syndrome. The social and psychological acceptance of the procedure is also discussed in terms of a developing country scenario. METHODS: A total of 14 patients with Mayer-Rokitansky-Kuster-Hauser syndrome were treated at our institute from January 1995 to December 2004. Sigmoid vaginoplasty was performed in all patients. The procedure was performed using a combined abdominoperineal approach. Dissection was done between the urethra and rectum to create a bed for the neovaginal colon conduit. A 10-cm segment of sigmoid colon was raised on its vascular pedicle, delivered through the abdominoperineal tunnel, and fixed to the vaginal pit incision. The patient records were reviewed for surgical technique and postoperative complications. Patients underwent a personal interview to assess the postoperative results, social acceptance of the procedure, and sexual satisfaction. RESULTS: The mean patient age at surgery was 16.8 years. The patients who underwent sigmoid vaginoplasty had good cosmetic results without the need for routine dilation or the problem of excessive mucus production. The postoperative morbidity was minimal. During a mean follow-up of 4.1 years, no stenosis or colitis was encountered. The subjective satisfaction rate with the surgical outcomes in all the patients was 8.01 on a scale of 0 to 10 (0, very disappointed to 10, satisfied). CONCLUSIONS: Sigmoid vaginoplasty is an effective treatment for patients with vaginal atresia. Timed vaginal reconstruction in these patients allows for a better quality of life and social acceptance. It also enables the patient to lead a near-normal sexual life, with high satisfaction rates.

A simple and rapid spectrophotometric method for determination of thiophanate-methyl in commercial formulations and its residues in foodstuffs.

A new, simple, rapid, and sensitive spectrophotometric method for the determination of thiophanate-methyl, based on its reaction with cobalt(II) in the presence of triethylamine, has been developed. The yellowish green color that develops instantaneously on mixing the fungicide with the reagents in dimethylformamide is stable for at least 2 h and has maximum absorbance at 360 nm. The method has been successfully applied to the determination of thiophanate-methyl in its commercial formulations and residues on grains and apples. A photometric titration procedure for formulation analysis of the fungicide has also been developed.