Complete Genome Sequencing Reveals Unusual Equine Rotavirus A of Bat Origin from India.

Rotaviruses are the most common viral agents associated with foal diarrhea. Between 2014 and 2017, the annual prevalence of rotavirus in diarrheic foals ranged between 18 and 28% in Haryana (India). Whole-genome sequencing of two equine rotavirus A (ERVA) isolates (RVA/Horse-wt/IND/ERV4/2017 and RVA/Horse-wt/IND/ERV6/2017) was carried out to determine the genotypic constellations (GCs) of ERVAs. The GCs of both the isolates were G3-P[3]-I8-R3-C3-M3-A9-N3-T3-E3-H6, a unique combination reported for ERVAs so far. Both the isolates carried VP6 of genotype I8, previously unreported from equines. Upon comparison with RVAs of other species, the GC of both isolates was identical to that of a bat rotavirus strain, MSLH14, isolated from China in 2012. The nucleotide sequences of the genes encoding VP3, NSP2, and NSP3 shared >95.81% identity with bat RVA strains isolated from Africa (Gabon). The genes encoding VP1, VP2, VP7, NSP1, and NSP4 shared 94.82% to 97.12% nucleotide identities with the human strains which have zoonotic links to bats (RCH272 and MS2015-1-0001). The VP6 genes of both strains were distinct and had the highest similarity of only 87.08% with that of CMH222, a human strain of bat origin. The phylogenetic analysis and lineage studies revealed that VP7 of both isolates clustered in a new lineage (lineage X) of the G3 genotype with bat, human, and alpaca strains. Similarly, VP4 clustered in a distinct P[3] lineage. These unusual findings highlight the terra incognita of the genomic diversity of equine rotaviruses and support the need for the surveillance of RVAs in animals and humans with a "one health" approach. IMPORTANCE Rotaviruses are globally prevalent diarrheal pathogens in young animals including foals, piglets, calves, goats, sheep, cats, and dogs along with humans. The genome of rotaviruses consists of 11 segments, which enables them to undergo reshuffling by reassortment of segments from multiple species during mixed infections. In this study, the prevalence of equine rotaviruses was 32.11% in organized equine farms of North India. The complete genome analysis of two ERVA isolates revealed an unusual genomic constellation, which was previously reported only in a bat RVA strain. A segment-wise phylogenetic analysis revealed that most segments of both isolates were highly similar either to bat or to bat-like human rotaviruses. The occurrence of unusual bat-like rotaviruses in equines emphasizes the need of extensive surveillance of complete genomes of both animal and human rotaviruses with a "one health" approach.

Genome editing (CRISPR-Cas)-mediated virus resistance in potato (Solanum tuberosum L.).

Plant viruses are the major pathogens that cause heavy yield loss in potato. The important viruses are potato virus X, potato virus Y and potato leaf roll virus around the world. Besides these three viruses, a novel tomato leaf curl New Delhi virus is serious in India. Conventional cum molecular breeding and transgenics approaches have been applied to develop virus resistant potato genotypes. But progress is slow in developing resistant varieties due to lack of host genes and long breeding process, and biosafety concern with transgenics. Hence, CRISPR-Cas mediated genome editing has emerged as a powerful technology to address these issues. CRISPR-Cas technology has been deployed in potato for several important traits. We highlight here CRISPR-Cas approaches of virus resistance through targeting viral genome (DNA or RNA), host factor gene and multiplexing of target genes simultaneously. Further, advancement in CRISPR-Cas research is presented in the area of DNA-free genome editing, virus-induced genome editing, and base editing. CRISPR-Cas delivery, transformation methods, and challenges in tetraploid potato and possible methods are also discussed.

Loop mediated isothermal amplification assay for detection of Trichomonas vaginalis in vaginal swabs among symptomatic women from North India.

Trichomonas vaginalis is one of the most common curable sexually transmitted pathogens infecting both men and women worldwide. Unlike traditional methods such as microscopy and culture, nucleic acid amplification tests rapidly detect this agent, assisting in treatment. Conventional polymerase chain reaction (PCR), the loop-mediated isothermal amplification (LAMP), and the Xpert TV assay were evaluated using 28 microscopy positive T. vaginalis samples and 125 microscopy negative samples from symptomatic females of reproductive age. The sensitivity of all tests was 100% and the specificity was 100%, 100%, and 99·2% for PCR, Xpert TV, and LAMP, respectively. The inter-rater reliability was excellent for PCR: Xpert TV (kappa-coefficient = 1) and good for LAMP assay: Xpert TV/PCR (kappa-coefficient = 0·98) and conventional PCR: LAMP (kappa-coefficient = 0·98). The study highlights the importance of PCR for screening T. vaginalis in women, particularly in laboratories where the Xpert-TV assay is not available or not affordable. The LAMP assay showed a lower positive predictive value which merits further evaluation. SIGNIFICANCE AND IMPACT OF THE STUDY: Trichomonas vaginalis is a common sexually transmitted pathogen associated with considerable morbidity and risk of complications. Due to the limitations of traditional diagnostic modalities, three molecular assays were compared: conventional polymerase chain reaction (PCR), Xpert TV assay, and loop mediated isothermal amplification (LAMP) assay for detecting T. vaginalis in symptomatic females. All tests had a sensitivity of 100% and the inter-rater reliability was excellent for PCR: Xpert TV, and good for LAMP assay: Xpert TV/PCR. The translational impact of this study lies in the possible use of conventional PCR and LAMP in laboratories where the Xpert TV assay is not available or not affordable.

Localized insulin amyloidosis with use of concentrated insulin: a potential complication.

BACKGROUND: Insulin-derived amyloidosis is a rare form of amyloidosis composed of insulin fibrils. The pH and concentration of insulin are known to influence the conformational state of the insulin hormone, with an increasing concentration favouring a more complex conformation. Concentrated insulin delivers a large amount of insulin to a localized area, raising the possibility of inducing conformational changes, forming insulin fibrils and leading to localized insulin amyloidosis. CASE REPORT: A middle-aged woman with long history of Type 2 diabetes mellitus, treated with concentrated human insulin (U-500 insulin) presented with nodular lesions at the site of her daily insulin injections. A punch biopsy of the nodules showed skin with dermal amyloidosis staining favourably with Congo Red stain. The amyloid tumours were resected and areas positive for Congo Red stain were sent for liquid chromatography tandem mass spectrometry, which showed a peptide profile consistent with amyloid insulin. CONCLUSION: Concentrated insulin was first introduced in 1952, however, it is only over the last two decades that it has been used increasingly, in congruence with the increasing incidence of obesity and diabetes mellitus seen in the USA. Only a few cases of insulin amyloidosis at the site of injection have been described in literature. With the increase in the use of insulin, this complication seems to be occurring more frequently. This is the first case report of a person with diabetes mellitus who developed localized insulin amyloidosis with the use of concentrated insulin, and points towards a potential complication of developing insulin amyloidosis with the use of concentrated insulin.

Robot-assisted total knee arthroplasty in haemophilic arthropathy.

INTRODUCTION: Execution of total knee arthroplasty (TKA) in end-stage haemophilic arthropathy is challenging because of soft tissue fibrosis, flexion contractures, poor quality of the bone, and the altered bony anatomy. Restoring the lower limb alignment and achieving range of motion (ROM) is difficult. Robots have been used in TKA to reduce the chances of malalignment and improve accuracy and precision. However, there has been no report in literature on use of robots for TKA in haemophilic arthropathy. The aim of this study was to evaluate whether robot-assisted TKA can be successfully carried out in haemophilic arthropathy and what precision could be obtained. METHODS: Thirty-two robot-assisted TKA were evaluated in 29 haemophilia patients. The mean follow up period was 5 years (range, 3-7 years). Hip-knee-ankle (HKA) axis, component angles, and radiographic loosening were evaluated. Clinically ROM, Knee Society scores (KSS) and SF-36 were assessed. RESULTS: The HKA axis was within a range of 0 ± 3° in 30 knees (93.8%). The alignment of the components also presented satisfying results. KSS were improved from 27.1 to 82.8 postoperatively (P < 0.001). The ROM was improved from 70.7 to 84.7 postoperatively (P = 0.006). Complications included early haematoma in three knees, heterotopic ossification in three knees, periprosthetic infection in two knees. CONCLUSIONS: Though robotic TKA gives excellent accuracy of lower limb and component alignment, expensive cost, additional preparation time, longer operation time with similar clinical results in haemophilic arthropathy should be concerned.

Nanostructured lipid carriers based nanogel for meloxicam delivery: mechanistic, in-vivo and stability evaluation.

AIM: Our investigation was aimed to investigate the potential suitability of meloxicam-loaded nanostructured lipid carriers (MLX-NLC) gel for topical application. MAIN METHODS: MLX-NLC gel was prepared and in vivo skin penetration ability of the NLC gel was evaluated using confocal laser scanning microscopy. We studied the effect of MLX-NLC gel on the changes in lipid profile of skin to get an insight into its skin penetration enhancement mechanism. Acetic acid induced writhing test was performed to evaluate the analgesic effect. Drug concentration-time profile of MLX in rat plasma and skin after topical and oral treatment with MLX-NLC gel and oral MLX-solution, respectively, was observed. MLX-NLC gel was subjected to primary skin irritation test, sub-acute dermal toxicity study. Storage stability of MLX-NLC gel was also assessed for 90 days. KEY FINDINGS: NLC gel was effective in permeating Rhodamine 123 to deeper layers of rat skin. Changes in skin lipid prolife were observed in the rat skin on treatment with MLX-NLC gel and the results supported skin lipid extraction as a possible penetration enhancement mechanism. MLX-NLC gel demonstrated sustained pain inhibitory effect. Pharmacokinetics study established that topical application of MLX-NLC gel had the potential to avoid systemic uptake and hence the risk of systemic adverse effects. MLX-NLC gel demonstrated good skin tolerability and biosafety. Excellent physical stability of nanogel was observed at 4 ± 2 °C. SIGNIFICANCE: The study revealed that NLC gel is a promising carrier system for the topical application of MLX without side effects.

Efficacy and safety using autotransfusion system with postoperative shed blood following total knee arthroplasty in haemophilia.

The purpose of this study was to evaluate the efficacy and safety of postoperative wound drain salvage and autotransfusion system in haemophilic patients undergoing elective total knee arthroplasty (TKA). No literature exists on reinfusing drained blood in patient with haemophilia undergoing TKA. Eighty-eight knees of 66 patients received cemented TKA due to end-stage haemophilic arthropathy (group I; with autotransfusion in 59 knees, group II; without autotransfusion in 29 knees). In group I, the postoperative shed blood was transfused within 6 h after surgery. The amount of blood drainage and reinfused blood, rate and amount of allogenic transfusion, postoperative change of haemoglobin level, prothrombin time (PT) and activated partial thromboplastin time were analysed. The mean postoperative blood drainage was 932 ± 479 mL in group I and 830 ± 492 mL in group II (P > 0.05). The mean volume of blood reinfused was 530 ± 265 mL in group I. Allogenic transfusion was needed in six knees (10.2%) of group I and eight knees (27.6%) of group II (P = 0.036). The mean volume of allogenic transfusion was 480 ± 49 mL in group I and 1041 ± 691 mL in group II (P > 0.05). Changes of all the laboratory results before and after TKA showed no statistically significant difference except PT was prolonged in group I (P = 0.008) at postoperative day 1. Moreover, there was no significant complication related to either reinfusion or allogenic transfusion in both groups. This study showed that reinfusion of drained blood is a simple, safe and efficacious method in patients with haemophilia undergoing TKA.

Priorities, opportunities, and challenges for integrating microorganisms into Earth system models for climate change prediction.

Climate change jeopardizes human health, global biodiversity, and sustainability of the biosphere. To make reliable predictions about climate change, scientists use Earth system models (ESMs) that integrate physical, chemical, and biological processes occurring on land, the oceans, and the atmosphere. Although critical for catalyzing coupled biogeochemical processes, microorganisms have traditionally been left out of ESMs. Here, we generate a "top 10" list of priorities, opportunities, and challenges for the explicit integration of microorganisms into ESMs. We discuss the need for coarse-graining microbial information into functionally relevant categories, as well as the capacity for microorganisms to rapidly evolve in response to climate-change drivers. Microbiologists are uniquely positioned to collect novel and valuable information necessary for next-generation ESMs, but this requires data harmonization and transdisciplinary collaboration to effectively guide adaptation strategies and mitigation policy.

Computer-navigated total knee arthroplasty in haemophilic arthropathy.

Total knee arthroplasty (TKA) in end-stage haemophilic arthropathy is complex and challenging due to the altered bony anatomy, arthrofibrosis and muscle contractures. Computer navigation is especially advocated in patients with deformity or altered anatomy to improve alignment and to assist in ligament balancing. The objective of this study was to evaluate the results of computer-navigated TKA in haemophilic arthropathy. A consecutive series of computer-assisted TKA for the end-stage haemophilic arthropathy between February 2007 and December 2009 were evaluated. A total of 27 TKA were performed in 25 patients. Pre- and postoperative full-length weight-bearing radiographs were assessed for the axial limb alignment. The orientation of the components was measured on anteroposterior radiographs. Clinically, Knee Society score and Short Form-36 were evaluated. The mechanical axis of the leg was within a range of ±3° varus/valgus in 92% of the TKA. The coronal alignment of the femoral and tibial components was within a range of ±3 degrees in 96% of the knees. The clinical outcomes were significantly improved after the operation. There were no complications specific to the computer navigation. Computer-navigated TKA helps in restoring the mechanical axis and improves accuracy of orientation of the components in patients with end-stage haemophilic arthropathy. Potential benefits in long-term outcome require further investigation.

Deciphering of seed Health of common food grains (wheat, rice) of North Eastern UP and Gurgaon Haryana, India.

The stored random samples of food seeds of wheat and rice (60 samples) were purchased from places of Eastern UP and Gurgaon district Haryana. Its moisture contents were estimated. The Mycological investigations of wheat seeds revealed presence of a total number of 16 fungal species viz., Alternaria alternata, Aspergillus candidus, Aspergillus flavus, A. niger, A. ochraceous, A. phoenicis, A. tamari, A. terreus, A. sydowi, Fusarium moniliforme, F. oxysporum F. solani, P. glabrum, Rhizopus nigricans, Trichoderma viride and Trichothecium roseum. While mycological analysis of rice seeds showed presence of 15 fungal species viz., Alternaria padwickii, A. oryzae, Curvularia lunata, Fusarium moniliforme, Aspergillus clavatus, A. flavus, A. niger, Cladosporium sp., Nigrospora oryzae, Alternaria tenuissima, Chaetomium globosum, F. solani, Microascus cirrosus, Helminthosporium oryzae, Pyricularia grisea. It also projected variation in presence of fungal species in blotter and agar plate method of analysis. In wheat Blotter method of analysis showed 16 fungal species while agar plate depicted 13 fungal species. In rice Agar plate method depicted presence of 15 fungal species while blotter method shows presence of 12 fungal species. The insect analysis revealed that wheat samples were infected with Tribolium castaneum. While rice seeds sample showed presence of insect Sitophilus oryzae. The investigations revealed that Aspergillus flavus, A. niger, Sitophilus oryzae and Tribolium castaneum caused reduction in seed weight loss, seed germination, carbohydrate and protein contents of common food grains (wheat, rice). It also revealed that randomly selected A. flavus isolate 1 of wheat showed higher potential of aflatoxin B1 production (1392.940 µg/l) while rice isolate 2 showed 1231.117 µg/l production.

A phase II study of metformin plus pemetrexed and carboplatin in patients with non-squamous non-small cell lung cancer (METALUNG).

Immune checkpoint inhibitors (ICIs) ± chemotherapy is the standard treatment for driver mutation-negative non-small cell lung cancer (NSCLC). However, accessibility to ICIs in LMICs is limited due to high cost, and platinum-based chemotherapy remains the mainstay of treatment. Metformin has anticancer properties, and studies suggest synergism between metformin and pemetrexed. Based on preclinical evidence, this combination may be more beneficial for STK11-mutated NSCLC, a subgroup, inherently resistant to ICIs. In this Simon two-stage, single-arm phase 2 trial, we investigated metformin with pemetrexed-carboplatin (PC) in patients with treatment-naive stage IV non-squamous NSCLC. The primary outcome was 6-month progression-free survival (PFS) rate. Secondary outcomes were safety, overall survival (OS), overall response rate (ORR), proportion of STK11 mutation, and effect of STK11 mutation on 6-month PFS rate. The study was terminated for futility after interim analysis. The median follow-up was 34.1 months. The 6-month PFS rate was 28% (95% CI 12.4-0.46). The median PFS and OS were 4.5 (95% CI 2.2-6.1) and 7.4 months (95% CI 5.3-15.3), respectively. The ORR was 72%. Gastrointestinal toxicities were the most common. No grade 4/5 toxicities were reported. Targeted sequencing was possible in nine cases. Two patients had STK11 mutation and a poor outcome (PFS < 12 weeks). We could not demonstrate the benefit of metformin with CP in terms of improvement in 6-month PFS rate; however, the combination was safe (CTRI/2019/02/017815).

Differences in consumption of food items between obese and normal-weight people in India.

BACKGROUND: There is a rising prevalence of obesity in India, and diet may be a major determinant of this. We aimed to assess differences in types and quantities of food items consumed by obese and normal-weight people in India. METHODS: Cross-sectional data of 7067 factory workers and their families were used from the Indian Migration Study, conducted in four cities across northern, central and southern India. Food frequency questionnaire data were used to compare the quantities of consumption of 184 food items between 287 obese (body mass index>30 kg/m2) and 1871 normalweight (body mass index 18.50-22.99 kg/m2) individuals, using t tests and ANCOVAs. Individuals with diabetes,hypertension and cardio-vascular disease were excluded. SPSS 16.0 was used for analysis. RESULTS: After adjusting for age, sex, location and socioeconomic status, obese individuals were found to eat significantly larger quantities of 11 food items compared with normalweight individuals. These included phulkas, chapatis/parathas/naan, plain dosa, mutton/chicken pulao/biryani, chicken fried/grilled, rasam, mixed vegetable sagu, vegetable raitha, honey,beetroot and bottlegourd (p< 0.01). Consumption of plain milk was higher among normal-weight than among obese individuals (p< 0.05). Consumption of some of these food items was also found to increase by socioeconomic status, decrease by age, and be higher among men relative to women. CONCLUSION: Obese individuals were found to consume larger quantities of certain food items compared with normal weight individuals. Interventions should aim at limiting overall food consumption among obese individuals.

Central nervous system infections caused by pathogenic free-living amoebae: An Indian perspective.

Pathogenic free-living amoebae (FLA), namely Acanthamoeba sp., Naegleria fowleri and Balamuthia mandrillaris are distributed worldwide. These neurotropic amoebae can cause fatal central nervous system (CNS) infections in humans. This review deals with the demographic characteristics, symptoms, diagnosis, and treatment outcomes of patients with CNS infections caused by FLA documented in India. There have been 42, 25, and 4 case reports of Acanthamoeba granulomatous amoebic encephalitis (GAE), N. fowleri primary amoebic meningoencephalitis (PAM), and B. mandrillaris meningoencephalitis (BAE), respectively. Overall, 17% of Acanthamoeba GAE patients and one of the four BAE patients had some form of immunosuppression, and more than half of the N. fowleri PAM cases had history of exposure to freshwater. Acanthamoeba GAE, PAM, and BAE were most commonly seen in males. Fever, headache, vomiting, seizures, and altered sensorium appear to be common symptoms in these patients. Some patients showed multiple lesions with edema, exudates or hydrocephalus in their brain CT/MRI. The cerebrospinal fluid (CSF) of these patients showed elevated protein and WBC levels. Direct microscopy of CSF was positive for amoebic trophozoites in 69% of Acanthamoeba GAE and 96% of PAM patients. One-fourth of the Acanthamoeba GAE and all the BAE patients were diagnosed only by histopathology following autopsy/biopsy samples. Twenty-one Acanthamoeba GAE survivors were treated with cotrimoxazole, rifampicin, and ketoconazole/amphotericin B, and all eleven PAM survivors were treated with amphotericin B alongside other drugs. A thorough search for these organisms in CNS samples is necessary to develop optimum treatment strategies.

Essential Oils as Alternative Promising Anti-Candidal Agents: Progress and Prospects.

Candida albicans is one of the main agents responsible for opportunistic pathogenic infections. The progressive emergence of fungal resistance to conventional antibiotics and its side effects, as well as treatment costs, are considered major limitations for antifungal drugs. It has drawn scientists' attention to the search for potential substitution and reliable therapeutic alternatives for the antifungal compounds from sources like medicinal plants, which contain numerous bioactive compounds such as essential oils. Essential oils (EO), apart from having lower toxicity and better biodegradability, are eco-friendly in nature as compared with conventional antibiotics. Furthermore, extracted essential oils have been reported to possess potent antimicrobial, antiinflammatory, and antioxidant properties that nominate them as promising natural candidates to combat numerous fungal ailments. Thus, the determination of antifungal efficacy of essential oil-bearing plants on Candida spp. will provide miscellaneous knowledge for future clinical studies that are required for the development of new formulations as alternative therapeutic agents to control the growth of Candida species. Therefore, this review summarizes the gist of major essential oils that have been investigated for their anti- Candida potential with some recommendations for further study.

Application of clove and dill oils as an alternative of salphos for chickpea food seed storage.

Mycological investigations of 25 samples of stored chickpea food seeds (Cicer arietinum L.) from grocery stores of Gurgaon and Gorakhpur revealed occurrence of seventeen fungal species belonging to genus viz., Alternaria, Aspergillus, Chaetomium, Colletotrichum, Curvularia, Fusarium, Penicillium, Rhizopus, Rhizoctonia, and Sclerotium. In these Aspergillus flavus, A. niger, Fusarium oxysporum had dominance in terms of per cent occurrence. Only one species of Bruchid (Callosobruchus chinensis L.) occurred in all the 25 samples. The biodeterioration of seeds inoculated with fungi: A. flavus, A. niger, F. oxysporum and the insect-C. chinensis, revealed their role in seed deterioration. For chickpea food seed protection essential oils were extracted from edible commodity(clove(Lavang and dill(sowa) leaf). Clove(Lavang) oil registered highest antifungal activity inhibiting (100%) mycelial growth of fungi, viz. species Aspergillus flavus, A. niger, Fusarium oxysporum at 300 ppm but was fungicidal at 400 ppm. Dill (Sowa) oil showed complete inhibition at 400 ppm and was fungicidal at 500 ppm. While mixture of both the oils (clove and dill) showed complete inhibition (100%) and fungicidal action at 400 ppm against the dominant fungi. The oils showed 100% insect repellent activity and were found fungicidal at 0.02 ml dose and also insecticidal. The mixture of oils was cidal at 0.02 ml dose. The mixture of oils showed a broad antifungal spectrum at 500 ppm while only 70-93% inhibitory activity at 300 ppm. The oils' mixture's activity was not affected by temp, storage and autoclaving up to 150 days. Oils physico-chemical properties were studied. GC-MS analysis of clove(Lavang) oil depicted major components: 75.63%eugenol while dill(sowa) leaf oil had 25.14% apiole. Formulation of Mixture of oils was more effective showing complete seed protection i.e.no growth of fungi and insects upto 150 days storage than salphos (150 days). While salphos controlled only maximum three fungi (A. terreus, C. dematium, F. moniliforme). The formulated oils mixture did not have any adverse effect on the chickpea seeds and increased their shelf life.

Dose-dense Paclitaxel and Carboplatin as Neoadjuvant Chemotherapy for Stage IIB/IIIA Non-small Cell Lung Cancer - A Phase II trial.

AIMS: The approach to potentially resectable non-small cell lung cancer (NSCLC) remains controversial. There is a benefit of neoadjuvant chemotherapy (NACT), but the ideal regimen is unknown. We evaluated the efficacy and safety of dose-dense NACT in potentially resectable NSCLC in this phase II trial. MATERIALS AND METHODS: Paclitaxel at 80 mg/m2 on days 1, 8 and 15 with AUC-6 carboplatin on day 1, 3 weekly for four cycles was evaluated as NACT. Patients with Eastern Cooperative Oncology Group performance status 0-2, stage IIB and IIIA (with only non-bulky N2 nodes) were included. The primary end point was the objective response rate. Secondary end points included toxicity, progression-free survival, recurrence-free survival, complete resection rate and overall survival. The relative dose intensity (RDI) was calculated to define tolerability (CTRI/2016/05/006916). RESULTS: In total, 37 patients were enrolled (median age 55 years). Most (78.8%) were smokers. Most patients had adenocarcinoma (57.6%) and stage IIIA disease (81.0%) according to the seventh American Joint Committee on Cancer staging system. Seventy-eight per cent of patients completed four cycles. The objective response rate was 75.6% with a complete response in 10.8%. The mean RDI of paclitaxel was 88.61%, with 68.0% of patients able to maintain an RDI ≥85.0%. In total, 187 toxicity events were recorded (120 grade 1, 64 grade 2 and three grade 3 events). Common toxicities were peripheral neuropathy (20.3%), myalgia (19.8%), nausea (15.7%) and neutropenia (10.2%). There were no treatment-related deaths. Seventeen patients underwent surgery (lobectomy 82.4%). After a median follow-up of 47 months (95% confidence interval 27-50.7 months), the median progression-free survival was 9.6 months (7.4-17.4) and overall survival was 29.2 months (16.0-37.2). CONCLUSION: Dose-dense paclitaxel-carboplatin is feasible, safe and efficacious and should be evaluated further in potentially resectable NSCLC.

Australian experience with total pancreatectomy with islet autotransplantation to treat chronic pancreatitis.

BACKGROUND: This study aimed to describe the clinical outcomes of total pancreatectomy with islet autotransplantation (TP-IAT) in Australia. METHODS: Individuals selected for TP-IAT surgery according to the Minnesota Criteria (Appendix) without evidence of diabetes were evaluated including time to transplantation from pancreatectomy, islet numbers infused and post-transplantation HbA1c, C-peptide, total daily insulin and analgesic requirement. RESULTS: Sixteen individuals underwent TP-IAT from Australia and New Zealand between 2010 and 2020. Two recipients are deceased. The median islet equivalents/kg infused was 4244 (interquartile range (IQR) 2290-7300). The median C-peptide 1 month post-TP-IAT was 384 (IQR 210-579) pmol/L and at median 29.5 (IQR 14.5-46.5) months from transplant was 395 (IQR 139-862) pmol/L. Insulin independence was achieved in eight of 15 (53.3%) surviving recipients. A higher islet equivalents transplanted was most strongly associated with the likelihood of insulin independence (P < 0.05). Of the 15 surviving recipients, 14 demonstrated substantial reduction in analgesic requirement. CONCLUSION: The TP-IAT programme in Australia has been a successful new therapy for the management of individuals with chronic pancreatitis including hereditary forms refractory to medical treatment to improve pain management with 50% insulin independence rates.

Cloning and expression of chicken granulocyte-macrophage colony stimulating factor (GMCSF) gene.

Granulocyte-macrophage colony stimulating factor (GMCSF), a multifunctional cytokine can enhance immune responses when administered along with DNA vaccine. Aim of the present study was to clone and express the chicken GMCSF cytokine for use as 'genetic adjuvant'. Chicken GMCSF gene 435bp was amplified using specific primers in which restriction sites of BamHI and HindIII were at forward and reverse primers respectively. The PCR product was cloned into eukaryotic expression vector pcDNA 3.1(+) and clones were confirmed by restriction digestion and nucleotide sequencing. Functional activity of recombinant GMCSF was checked by expression of GMCSF specific mRNA in transfected Vero cells by RT-PCR of total RNA isolated from transfected Vero cells. The recombinant plasmid can be used as genetic adjuvant in chicken.

Re-emergence of glanders in India - Report of Maharashtra state.

Glanders, a notifiable highly contagious disease primarily of equids, is a disease of high zoonotic importance. Caused by gram-negative bacillus, Burkholderia mallei, the disease was restricted to certain pockets of India with sporadic cases. Recently, a major outbreak of glanders occurred in India starting from Maharashtra. Following clinical signs and symptoms and laboratory investigations on serum, nasal swab and pus swab samples, it was confirmed as glanders among equines in Pune and Panchgani areas of Maharashtra. One pus sample and three nasal swabs yielded B. mallei isolates while 23 serum samples were found positive for glanders by complement fixation test (CFT). The disease was successfully controlled in the state by following strategies for prevention of spread of the disease to other areas in accordance with Glanders and Farcy Act, 1899. Follow up of the occurrence in Maharashtra revealed negative status based on testing and physical surveillance on more than 3,500 equines thereafter. Investigations indicated that the nidus of infection may be present elsewhere in North India.

The Role of DNMT and HDACs in the Fetal Programming of Hypertension by Glucocorticoids.

The causes of hypertension are complex and involve both genetic and environmental factors. Environment changes during fetal development have been linked to adult diseases including hypertension. Studies show that timed in utero exposure to the synthetic glucocorticoid (GC) dexamethasone (Dex) results in the development of hypertension in adult rats. Evidence suggests that in utero stress can alter patterns of gene expression, possibly a result of alterations in the topology of the genome by epigenetic markers such as DNA methyltransferases (DNMTs) and histone deacetylases (HDACs). The objective of this study was to determine the effects of epigenetic regulators in the fetal programming and the development of adult hypertension. Specifically, this research examined the effects of the HDAC inhibitor valproic acid (VPA) and the DNMT inhibitor 5-aza-2'-deoxycytidine (5aza2DC) on blood pressure (BP) and gene expression in prenatal Dex-programmed rats. Data suggest that both VPA and 5aza2DC attenuated the Dex-mediated development of hypertension and restored BP to control levels. Epigenetic DNMT inhibition (DNMTi) or HDAC inhibition (HDACi) also successfully attenuated elevations in the majority of altered catecholamine (CA) enzyme expression, phenylethanolamine N-methyltransferase (PNMT) protein, and elevated epinephrine (Epi) levels in males. Although females responded to HDACi similar to males, DNMTi drove increased glucocorticoid receptor (GR) and PNMT expression and elevations in circulating Epi in females despite showing normotensive BP.