Flavin-containing monooxygenase : metabolism and toxicity of anti-tuberculosis drug ethionamide in lung

Multiple drug resistance (MDR) Tuberculosis (TB), leads to increased use of “second-line” drugs; one of the most effective is ethionamide (ETA). ETA is a prodrug metabolized by a mycobacterial flavin-containing monooxygenase (EtaA) as well human flavin-containing monooxygenases (FMOs). Of the five functional FMOs of humans, FMOs 1, 2, and 3 are the most important in metabolism of xenobiotics. FMO2 is a major isoform in the lung expressed in most mammals. In humans a polymorphism encoding inactive FMO2 is the common allele, while the wild type allele encoding active FMO2.1 has been documented only in individuals of African and Hispanic origin, at an incidence of up to 50% and 7%, respectively. Human FMO2.1 is more efficient than EtaA in the first S-oxygenation of ETA to ETA S-oxide (ETASO). ETASO is a sulfenic acid and capable of redox-cycling with glutathione (GSH) producing oxidative stress and toxicity. We hypothesized that the FMO2 polymorphism will result in differences in the concentration and spectrum of ETA prodrug and metabolites. TB-infected individuals expressing active FMO2.1 will have reduced ETA efficacy in inhibition and killing of M. tuberculosis and enhanced oxidative stress, pulmonary toxicity and cell death in the host. To test this hypothesis, Fmo1/2/4 knockout mice modeling the FMO2*2 genotype were utilized to study ETA metabolism and toxicity compared to wild type mice in order to model humans expressing the FMO2*1 allele. All mice were capable of metabolizing ETA to ETASO. Wild type mice had significantly higher plasma and epithelial lining fluid (ELF) levels of ETASO than ETA. In contrast, Fmo 1/2/4 knockout mice had higher plasma and ELF levels of ETA than ETASO. ETASO, a sulfenic acid, was significantly higher in wild type mice than knockout mice in both plasma and ELF. Investigation of the effects of higher levels of ETASO in wild type mice was warranted. Long-term ETA dosing was utilized to examine oxidative stress and toxicity. Compare to Fmo 1/2/4 knockout mice, wild type male and female mice showed more differentially expressed genes in lung related to oxidative stress and antioxidant defense pathways. Altered Fmo1, 2 and 4 expression was also found in lung and liver of long-term ETA-treated wild type mice in comparison to vehicle controls. To examine ETA therapeutic efficacy in the presence and absence of FMO, wild type and knockout mice were infected with M. avium and dosed with 50 mg/kg of ETA for 28-days and compared with vehicle-dosed mice for bactericidal effects. An ETA dose of 50 mg/kg did not significantly reduce the mycobacterium infection, leaving the fundamentally question of the role of FMO2 polymorphism on ETA efficacy unresolved. Though interestingly, significant higher bacterial load was observed in the ETA treated wildtype compared to ETA treated knockout mice which suggest that the presence of FMO in wildtype is making these mice more susceptible to bacterial infection when ETA is dosed. Collectively these studies suggest that humans expressing active FMO2.1 may be at higher risk of ETA toxicity than individuals with inactive FMO2.2 that are undergoing long-term ETA treatment for MDR-TB. This work highlights the potential of the FMO2 human genetic on TB drug selection to maximize treatment efficacy and minimize toxicity. Another study of M. tuberculosis infected mice with 125 mg/kg dose of ETA is required to understand ETA therapeutic efficacy in the presence and absence of FMOs.Keywords: ethionamide, ET

MicroRNA regulation of type 2 innate lymphoid cell homeostasis and function in allergic inflammation.

MicroRNAs (miRNAs) exert powerful effects on immunity through coordinate regulation of multiple target genes in a wide variety of cells. Type 2 innate lymphoid cells (ILC2s) are tissue sentinel mediators of allergic inflammation. We established the physiological requirements for miRNAs in ILC2 homeostasis and immune function and compared the global miRNA repertoire of resting and activated ILC2s and T helper type 2 (TH2) cells. After exposure to the natural allergen papain, mice selectively lacking the miR-17∼92 cluster in ILC2s displayed reduced lung inflammation. Moreover, miR-17∼92-deficient ILC2s exhibited defective growth and cytokine expression in response to IL-33 and thymic stromal lymphopoietin in vitro. The miR-17∼92 cluster member miR-19a promoted IL-13 and IL-5 production and inhibited expression of several targets, including SOCS1 and A20, signaling inhibitors that limit IL-13 and IL-5 production. These findings establish miRNAs as important regulators of ILC2 biology, reveal overlapping but nonidentical miRNA-regulated gene expression networks in ILC2s and TH2 cells, and reinforce the therapeutic potential of targeting miR-19 to alleviate pathogenic allergic responses

Robust Anti‐Tumor T Cell Response with Efficient Intratumoral Infiltration by Nanodisc Cancer Immunotherapy

Potent anti‐tumor T cell response and efficient intratumoral T cell infiltration are the major challenges for therapeutic cancer vaccines. To address these issues, a nanovaccine system is designed to promote anti‐tumor T cell responses, and intratumoral infiltration is examined in various murine tumor models. Subcutaneous vaccination with nanodiscs carrying human papillomavirus (HPV)‐16 E7 antigen elicits as high as ∼32% E7‐specific CD8α+ T cell responses in circulation, representing a 29‐fold improvement over the soluble peptide vaccination. Importantly, nanodisc vaccination also promotes robust intratumoral T cell infiltration and eliminates HPV16 E6/E7‐expressing TC‐1 tumors at mucosal sites, including lungs, inner lip, and intravaginal tissues. In a benchmark study with a live Listeria vaccine combined with anti‐PD‐1 IgG, nanodiscs plus anti‐PD‐1 immune checkpoint blockade elicits comparable levels of T cell responses with anti‐tumor efficacy. Furthermore, compared with Complete Freund’s Adjuvant combined with tetanus toxoid, nanodisc vaccination in HLA‐A02 mice generates >200‐fold stronger IFN‐γ+ T cell responses against a neoantigen from an HLA‐A02 melanoma patient. Overall, these results show that the nanodisc system is a promising cancer vaccine platform for inducing anti‐tumor T cell responses.Efficient infiltration of T cells in solid cancer is a major challenge for cancer immunotherapy. A nanoparticle vaccine system is developed to promote T cell infiltration into peripheral mucosal tissues and eliminate disseminated tumors. Nanodiscs are broadly applicable with a wide range of tumor antigens, thus providing a versatile and potent vaccine platform for eliciting T cell immunity.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/156420/3/adtp202000094.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156420/2/adtp202000094-sup-0001-SuppMat.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156420/1/adtp202000094_am.pd

Burden of malaria in pregnancy in Jharkhand State, India

<p>Abstract</p> <p>Background</p> <p>Past studies in India included only symptomatic pregnant women and thus may have overestimated the proportion of women with malaria. Given the large population at risk, a cross sectional study was conducted in order to better define the burden of malaria in pregnancy in Jharkhand, a malaria-endemic state in central-east India.</p> <p>Methods</p> <p>Cross-sectional surveys at antenatal clinics and delivery units were performed over a 12-month period at two district hospitals in urban and semi-urban areas, and a rural mission hospital. Malaria was diagnosed by Giemsa-stained blood smear and/or rapid diagnostic test using peripheral or placental blood.</p> <p>Results</p> <p>2,386 pregnant women were enrolled at the antenatal clinics and 718 at the delivery units. 1.8% (43/2382) of the antenatal clinic cohort had a positive diagnostic test for malaria (53.5% <it>Plasmodium falciparum</it>, 37.2% <it>Plasmodium vivax</it>, and 9.3% mixed infections). Peripheral parasitaemia was more common in pregnant women attending antenatal clinics in rural sites (adjusted relative risk [aRR] 4.31, 95%CI 1.84-10.11) and in those who were younger than 20 years (aRR 2.68, 95%CI 1.03-6.98). Among delivery unit participants, 1.7% (12/717) had peripheral parasitaemia and 2.4% (17/712) had placental parasitaemia. Women attending delivery units were more likely to be parasitaemic if they were in their first or second pregnancy (aRR 3.17, 95%CI 1.32-7.61), had fever in the last week (aRR 5.34, 95%CI 2.89-9.90), or had rural residence (aRR 3.10, 95%CI 1.66-5.79). Malaria control measures including indoor residual spraying (IRS) and untreated bed nets were common, whereas insecticide-treated bed nets (ITN) and malaria chemoprophylaxis were rarely used.</p> <p>Conclusion</p> <p>The prevalence of malaria among pregnant women was relatively low. However, given the large at-risk population in this malaria-endemic region of India, there is a need to enhance ITN availability and use for prevention of malaria in pregnancy, and to improve case management of symptomatic pregnant women.</p

Sterile Protection against Plasmodium knowlesi in Rhesus Monkeys from a Malaria Vaccine: Comparison of Heterologous Prime Boost Strategies

Using newer vaccine platforms which have been effective against malaria in rodent models, we tested five immunization regimens against Plasmodium knowlesi in rhesus monkeys. All vaccines included the same four P. knowlesi antigens: the pre-erythrocytic antigens CSP, SSP2, and erythrocytic antigens AMA1, MSP1. We used four vaccine platforms for prime or boost vaccinations: plasmids (DNA), alphavirus replicons (VRP), attenuated adenovirus serotype 5 (Ad), or attenuated poxvirus (Pox). These four platforms combined to produce five different prime/boost vaccine regimens: Pox alone, VRP/Pox, VRP/Ad, Ad/Pox, and DNA/Pox. Five rhesus monkeys were immunized with each regimen, and five Control monkeys received a mock vaccination. The time to complete vaccinations was 420 days. All monkeys were challenged twice with 100 P. knowlesi sporozoites given IV. The first challenge was given 12 days after the last vaccination, and the monkeys receiving the DNA/Pox vaccine were the best protected, with 3/5 monkeys sterilely protected and 1/5 monkeys that self-cured its parasitemia. There was no protection in monkeys that received Pox malaria vaccine alone without previous priming. The second sporozoite challenge was given 4 months after the first. All 4 monkeys that were protected in the first challenge developed malaria in the second challenge. DNA, VRP and Ad5 vaccines all primed monkeys for strong immune responses after the Pox boost. We discuss the high level but short duration of protection in this experiment and the possible benefits of the long interval between prime and boost

Fatty Acid Composition of Developing Sea Buckthorn (Hippophae rhamnoides L.) Berry and the Transcriptome of the Mature Seed

Background: Sea buckthorn (Hippophae rhamnoides L.) is a hardy, fruit-producing plant known historically for its medicinal and nutraceutical properties. The most recognized product of sea buckthorn is its fruit oil, composed of seed oil that is rich in essential fatty acids, linoleic (18:2\u3c9-6) and \u3b1-linolenic (18:3\u3c9-3) acids, and pulp oil that contains high levels of monounsaturated palmitoleic acid (16:1\u3c9-7). Sea buckthorn is fast gaining popularity as a source of functional food and nutraceuticals, but currently has few genomic resources; therefore, we explored the fatty acid composition of Canadian-grown cultivars (ssp. mongolica) and the sea buckthorn seed transcriptome using the 454 GS FLX sequencing technology. Results: GC-MS profiling of fatty acids in seeds and pulp of berries indicated that the seed oil contained linoleic and \u3b1-linolenic acids at 33-36% and 30-36%, respectively, while the pulp oil contained palmitoleic acid at 32-42%. 454 sequencing of sea buckthorn cDNA collections from mature seeds yielded 500,392 sequence reads, which identified 89,141 putative unigenes represented by 37,482 contigs and 51,659 singletons. Functional annotation by Gene Ontology and computational prediction of metabolic pathways indicated that primary metabolism (protein>nucleic acid>carbohydrate>lipid) and fatty acid and lipid biosynthesis pathways were highly represented categories. Sea buckthorn sequences related to fatty acid biosynthesis genes in Arabidopsis were identified, and a subset of these was examined for transcript expression at four developing stages of the berry. Conclusion: This study provides the first comprehensive genomic resources represented by expressed sequences for sea buckthorn, and demonstrates that the seed oil of Canadian-grown sea buckthorn cultivars contains high levels of linoleic acid and \u3b1-linolenic acid in a close to 1:1 ratio, which is beneficial for human health. These data provide the foundation for further studies on sea buckthorn oil, the enzymes involved in its biosynthesis, and the genes involved in the general hardiness of sea buckthorn against environmental conditions.Peer reviewed: YesNRC publication: Ye

Shifting the limits in wheat research and breeding using a fully annotated reference genome

Introduction: Wheat (Triticum aestivum L.) is the most widely cultivated crop on Earth, contributing about a fifth of the total calories consumed by humans. Consequently, wheat yields and production affect the global economy, and failed harvests can lead to social unrest. Breeders continuously strive to develop improved varieties by fine-tuning genetically complex yield and end-use quality parameters while maintaining stable yields and adapting the crop to regionally specific biotic and abiotic stresses. Rationale: Breeding efforts are limited by insufficient knowledge and understanding of wheat biology and the molecular basis of central agronomic traits. To meet the demands of human population growth, there is an urgent need for wheat research and breeding to accelerate genetic gain as well as to increase and protect wheat yield and quality traits. In other plant and animal species, access to a fully annotated and ordered genome sequence, including regulatory sequences and genome-diversity information, has promoted the development of systematic and more time-efficient approaches for the selection and understanding of important traits. Wheat has lagged behind, primarily owing to the challenges of assembling a genome that is more than five times as large as the human genome, polyploid, and complex, containing more than 85% repetitive DNA. To provide a foundation for improvement through molecular breeding, in 2005, the International Wheat Genome Sequencing Consortium set out to deliver a high-quality annotated reference genome sequence of bread wheat. Results: An annotated reference sequence representing the hexaploid bread wheat genome in the form of 21 chromosome-like sequence assemblies has now been delivered, giving access to 107,891 high-confidence genes, including their genomic context of regulatory sequences. This assembly enabled the discovery of tissue- and developmental stage–related gene coexpression networks using a transcriptome atlas representing all stages of wheat development. The dynamics of change in complex gene families involved in environmental adaptation and end-use quality were revealed at subgenome resolution and contextualized to known agronomic single-gene or quantitative trait loci. Aspects of the future value of the annotated assembly for molecular breeding and research were exemplarily illustrated by resolving the genetic basis of a quantitative trait locus conferring resistance to abiotic stress and insect damage as well as by serving as the basis for genome editing of the flowering-time trait. Conclusion: This annotated reference sequence of wheat is a resource that can now drive disruptive innovation in wheat improvement, as this community resource establishes the foundation for accelerating wheat research and application through improved understanding of wheat biology and genomics-assisted breeding. Importantly, the bioinformatics capacity developed for model-organism genomes will facilitate a better understanding of the wheat genome as a result of the high-quality chromosome-based genome assembly. By necessity, breeders work with the genome at the whole chromosome level, as each new cross involves the modification of genome-wide gene networks that control the expression of complex traits such as yield. With the annotated and ordered reference genome sequence in place, researchers and breeders can now easily access sequence-level information to precisely define the necessary changes in the genomes for breeding programs. This will be realized through the implementation of new DNA marker platforms and targeted breeding technologies, including genome editing

A study of prevalence of Anisometropia and associated amblyopia in patients with refractive error

Aims: The present study was planned to evaluate the prevalence of anisometropia in patients with refractive error, to compare the prevalence of different types of anisometropia and to study the prevalence of amblyopia in anisometropic patients. Material and Method: It was a cross sectional study in which 400 randomly selected patients were evaluated for anisometropia and also for anisometropic amblyopia. All patients underwent thorough ophthalmic examination including visual acuity testing, slit lamp examination and fundus examination. Later on the data was analysed statistically with different formulas like Chi square test, Fisher exact test etc. Results: The prevalence of anisometropia was found to be 16.5% in total 400 patients with refractive errors, out of which 57.5% were females and 42.5% were male patients. Prevalence was found maximum in 20-30 years age group (36.4%). Highest prevalence among various types of anisometropia was found of compound myopic anisometropia (37.9%). Conclusion: No gender difference was seen in prevalence of anisometropia. Prevalence of anisometropia significantly varied with different age groups. The prevalence of amblyopia in patients of anisometropia was found to be statistically significant (p value=.0001)

Prevalence of alcohol and drug dependence in rural and slum population of Chandigarh: A community survey

The present epidemiological survey was conducted by the department of psychiatry, Govt. Medical College and Hospital, Chandigarh to estimate the pattern of alcohol and other substance dependence in rural and slum dwellers population of Chandigarh. In this survey 6.88% individuals of the total population surveyed (2992) fulfilled dependence criteria of ICD-10. Alcohol was the primary substance of dependence for majority of urban slum substance users and rural areas users. Age at first drug use was 20.89 ± 5.31 years (mean ± S.D) among rural population and 19.75 ± 5.4 years (mean ± SD) in urban slums. Majority of them reported having health related complications (85.71%) followed by family problems (77.31%) due to drug dependence. This survey reflects the need to intensify efforts at the community level to reach the unreached