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The expression of ATP-binding cassette transporter (ABC transporters) has been reported in various tissues such as the lung, liver, kidney, brain and intestine. These proteins account for the efflux of different compounds and metabolites across the membrane, thus decreasing the concentration of the toxic compounds. ABC transporter genes play a vital role in the development of multidrug resistance, which is the main obstacle that hinders the success of chemotherapy. Preclinical and clinical trials have investigated the probability of overcoming drug-associated resistance and substantial toxicities. The focus has been put on several strategies to overcome multidrug resistance. These strategies include the development of modulators that can modulate ABC transporters. This knowledge can be translated for clinical oncology treatment in the future.

The present research endeavour was executed with 59 rye introgressed wheat lines including 52 triticale × wheat stable lines and 7 wheat × rye (Lahaul Local) introgressed doubled haploids. The detection of new recombinants through molecular cytogenetic tools viz. genomic in situ hybridization and fluorescence in situ hybridization revealed 1BL.1RS translocation in 33 lines, 1BL.1RS translocation along with substitution in 5 lines, 1BL.1RS translocation along with addition in 8 lines, substitution of single pair of rye chromosomes in 5 lines and addition of rye chromatin in 1 line, while 7 lines were found with no rye chromatin. To validate the effect of rye introgression on drought tolerance and yellow rust resistance, all 59 recombinants were evaluated under well-watered and water stressed conditions at in vivo. Analysis of variance revealed sufficient genetic variability among genotypes for different traits at both in vivo level. Under field conditions, direct influence of rye introgression was observed on plant height, grain yield, biological yield and harvest index. Overall, 1BL.1RS translocations and substitutions were found to exhibit significant influence on most of the drought parameters. The screening for yellow rust resistance revealed differential disease reaction, where 52 recombinants were observed to be resistant. The lines exhibiting superiority for the traits can be used as a parental source in wheat improvement endeavours.

SARS-CoV-2 remains a global threat to human health particularly as escape mutants emerge. There is an unmet need for effective treatments against COVID-19 for which neutralizing single domain antibodies (nanobodies) have significant potential. Their small size and stability mean that nanobodies are compatible with respiratory administration. We report four nanobodies (C5, H3, C1, F2) engineered as homotrimers with pmolar affinity for the receptor binding domain (RBD) of the SARS-CoV-2 spike protein. Crystal structures show C5 and H3 overlap the ACE2 epitope, whilst C1 and F2 bind to a different epitope. Cryo Electron Microscopy shows C5 binding results in an all down arrangement of the Spike protein. C1, H3 and C5 all neutralize the Victoria strain, and the highly transmissible Alpha (B.1.1.7 first identified in Kent, UK) strain and C1 also neutralizes the Beta (B.1.35, first identified in South Africa). Administration of C5-trimer via the respiratory route showed potent therapeutic efficacy in the Syrian hamster model of COVID-19 and separately, effective prophylaxis. The molecule was similarly potent by intraperitoneal injection., Neutralizing nanobodies (Nb) are of considerable interest as therapeutic agents for COVID-19 treatment. Here, the authors functionally and structurally characterize Nbs that bind with high affinity to the receptor binding domain of the SARS-CoV-2 spike protein and show that an engineered homotrimeric Nb prevents disease progression in a Syrian hamster model of COVID-19 when administered intranasally.

The ameliorative effect of seed priming with salicylic acid (SA) on Cr6+ and Zn2+ induced stress responses in wheat (Triticum aestivum L.) was evaluated using biochemical and computational approaches. The results suggested that Cr6+ and Zn2+ stress in wheat seedlings in absence of SA priming resulted in significant inhibition of growth, biomass loss with high accumulation of reactive oxygen species (ROS) leading to oxidative stress and inactivation of antioxidant metabolism. Wheat seeds primed with SA ameliorated the effects of Cr6+ and Zn2+ induced stress by maintaining appropriate growth and biomass of the seedlings. Moreover, when compared to the SA non-primed wheat seedlings, the levels of ROS and oxidative stress load were low in SA-primed seedlings subjected to Cr6+ and Zn2+ stress, which clearly indicated an ameliorative effect of seed priming with SA. Though, significant variation of antioxidant enzymes such as catalase (CAT) and superoxide dismutase (SOD) was not observed in wheat seedlings under stress conditions, and the non-enzymatic antioxidants such as ascorbic acid (AsA) and reduced glutathione (GSH) showed higher levels in SA-primed seedlings as compared to non-primed groups. The computational (molecular docking) analysis with Cu–Zn–SOD, Mn–SOD and CAT as target enzymes revealed that SA may potentially interfere with the binding and interaction of Cr6+ and Zn2+ ions with the active site of Cu–Zn–SOD effectively. Though, this computational analysis is based on structures of the targets derived through homology modeling, nevertheless it fundamentally substantiated our experimental findings on CAT and SOD activities.

The post-genomic era has witnessed several new possibilities to understand diverse functional aspects of plants quite precisely. From genomics to metabolomics and now phenomics, the complex interplay of these biological networks has been successfully elucidated. Abiotic stresses, such as drought, flooding, exposure to heavy metals and metalloids, and high or low temperature are foremost constraints in agriculture, and remains as the major reason for poor crop productivity and low yield globally. The primary aim of metabolomics is to identify final gene products, the metabolites, which serve as prospective markers (or traits) to comprehend abiotic stress adaptation and tolerance in plants. This review provides an overview on the application of metabolomics as a comprehensive tool for “Systems Biology Approach” to unravel the complex interaction of networks and components in plants towards abiotic stress adaptation and tolerance.

Extensive use of simple sequence repeat (SSR) is facilitated if loci would be transferable across species even in closely related genera to overcome high cost and efforts involved in their development as major constraints. In the present study, apple and pear genomic microsatellite primer pairs were used to amplify SSR loci in apple, pear, quince and loquat genotypes, respectively. Already reported SSRs were selected based on their polymorphic survey for successful amplification with at least one polymerase chain reaction (PCR) product of the approximate size expected for a homologous locus screened among apple and pear genotypes for further transferability exploration across other temperate pome fruit crops, respectively. Highest transferability of apple and pear SSR, 61.53 % and 73.33 % was observed in closely related quince and apple genotypes, respectively. This indicated that primer binding sites between these two closely related genera, Malus and Pyrus, are fairly well conserved. Maximum transferability rate was found to be 93.33 % and 80.00 % across all the subjected genotypes for primer CH05D11 and TSUenh016 in apple and pear, respectively. The transferability of markers is based on genomic similarity, and can reflect the relationship of genome collinearity and even evolution between species. This high level of transferability of apple and pear SSRs to other temperate pome fruit crops indicated their promise for application to future molecular screening, map construction, and comparative genomic studies, etc.

Plant microbiota play essential roles in plant health and crop productivity. Comparisons of community composition have suggested seed, soil, and the atmosphere as reservoirs of phyllosphere microbiota. After finding that leaves of tomato (Solanum lycopersicum) plants exposed to rain carried a higher microbial population size than leaves of tomato plants not exposed to rain, we experimentally tested the hypothesis that rain is a thus-far-neglected reservoir of phyllosphere microbiota. Therefore, rain microbiota were compared with phyllosphere microbiota of tomato plants either treated with concentrated rain microbiota, filter-sterilized rain, or sterile water. Based on 16S ribosomal RNA amplicon sequencing, 104 operational taxonomic units (OTUs) significantly increased in relative abundance after inoculation with concentrated rain microbiota but no OTU significantly increased after treatment with either sterile water or filter-sterilized rain. Some of the genera to which these 104 OTUs belonged were also found at higher relative abundance on tomato plants exposed to rain outdoors than on tomato plants grown protected from rain in a commercial greenhouse. Taken together, these results point to precipitation as a reservoir of phyllosphere microbiota and show the potential of controlled experiments to investigate the role of different reservoirs in the assembly of phyllosphere microbiota.

To assess relative efficiency and identify efficient pollen source among maize and Imperata cylindrica for haploid induction, 21 maize and two Imperata cylindrica genotypes were hybridized as pollen parent with each of the four hexaploid, two tetraploid wheats and two genotypes of triticale following wheat × maize and wheat × Imperata cylindrica system of haploid induction. Observations for haploid induction parameters showed Imperata cylindrica as significantly superior pollen source than maize. Mean PFF, EFF, HRF and HFF obtained with Imperata were 55.05%, 24.07%, 18.58% and 4.96%, respectively, whereas the frequencies recorded with maize were 53.80%, 6.72%, 39.36% and 1.71%, respectively. Triticale remained nonresponsive to embryo formation with both the pollen species. The line × tester analysis showed significant effects of both male and female genotypes on all the haploid induction parameters. Analysis of GCA revealed differences among pollen sources in their potential to induce haploids. In maize, genotypes HPMC-57, HPMC-58, HPMC-25 for EFF and HPMC-58 for HFF emerged out to be best general combiners, while in the case of Imperata, GCA values were high for Ic-ye genotype. The present study put forth Imperata as an efficient pollen source over maize for haploid induction in hexaploid and tetraploid wheats.

Durum wheat × Imperata cylindrica hybridization was carried out utilizing two durum wheat genotypes as maternal parents (A-9-30-1 and PDW 314) and I. cylindrica as a pollen source followed by in vivo application of colchicine to establish a standard protocol for colchicine application aimed at enhancement of doubled haploid production in durum wheat. Various concentrations of colchicine ranging from 500 to 7000 ppm were injected to the pollinated spikes at different time interval viz., 24 h, 48 h and 72 h after pollination. Application of 5000–7000 ppm colchicine at 48 h after pollination was established as the optimum colchicine dose for chromosome doubling as confirmed through cytological studies in durum wheat using I. cylindrica-mediated chromosome elimination approach. PDW 314 was identified as a durum wheat genotype having more potential for doubled haploid induction via in vivo colchicine application than A-9-30-1. Identification of optimum dose of colchicine for application at in vivo level for induction of doubled haploids has opened new vistas in durum wheat improvement programme.

Coronavirus disease 2019 (COVID-19) is a primarily respiratory disease with variable clinical courses for which animal models are needed to gather insights into the pathogenesis of its causative virus, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), in human patients. SARS-CoV-2 not only affects the respiratory tract but also the central nervous system (CNS), leading to neurological symptoms such as loss of smell and taste, headache, fatigue or severe complications like cerebrovascular diseases. Transgenic mice expressing human angiotensin-converting enzyme 2 (hACE2) under the cytokeratin 18 promoter (K18-hACE2) represent a well-known model of SARS-CoV-2 infection. In the present study, it served to investigate the spatiotemporal distribution and pathomorphological features in the CNS following intranasal infection with relatively low SARS-CoV-2 doses and after prior influenza A virus infection. In K18-hACE2 mice, SARS-CoV-2 was found to frequently spread to and within the CNS during the later phase (day 7) of infection. Infection was restricted to neurons and appeared to first affect the olfactory bulb and spread from there mainly in basally orientated regions in the brain and into the spinal cord, in a dose dependent manner and independent of ACE2 expression. Neuronal infection was not associated with cell death, axonal damage or demyelination. However, microglial activation, microgliosis and a mild macrophage and T cell dominated inflammatory response was consistently observed. This was accompanied by apoptotic death of endothelial, microglial and immune cells, without evidence of viral infection of glial cells, endothelial cells and leukocytes. Taken together, microgliosis and immune cell apoptosis indicate a potential important role of microglial cells for the pathogenesis and viral effect in COVID-19 and possible impairment of neurological functions, especially in long COVID. These data may also be informative for the selection of therapeutic candidates, and broadly support investigation of agents with adequate penetration into relevant regions of the CNS.

Earth's radiation budget and frequency and intensity of precipitation are influenced by aerosols with ice nucleation activity (INA), i.e., particles that catalyze the formation of ice. Some bacteria, fungi, and pollen are among the most efficient ice nucleators but the molecular basis of INA is poorly understood in most of them. Lysinibacillus parviboronicapiens (Lp) was previously identified as the first Gram-positive bacterium with INA. INA of Lp is associated with a secreted, nanometer-sized, non-proteinaceous macromolecule or particle. Here a combination of comparative genomics, transcriptomics, and a mutant screen showed that INA in Lp depends on a type I iterative polyketide synthase and a non-ribosomal peptide synthetase (PKS-NRPS). Differential filtration in combination with gradient ultracentrifugation revealed that the product of the PKS-NRPS is associated with secreted particles of a density typical of extracellular vesicles and electron microscopy showed that these particles consist in "pearl chain"-like structures not resembling any other known bacterial structures. These findings expand our knowledge of biological INA, may be a model for INA in other organisms for which the molecular basis of INA is unknown, and present another step towards unraveling the role of microbes in atmospheric processes. National Science FoundationNational Science Foundation (NSF) [IOS-1754721]; Virginia Agricultural Experiment Station; Hatch Program of the National Institute of Food and Agriculture, US Department of Agriculture Published version This study was supported by the National Science Foundation (IOS-1754721). Funding to Boris A. Vinatzer was also provided in part by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, US Department of Agriculture. We would like to thank Kate Costello, Nakysa Kheirandish, Michelle Zambrano, and Nnanna Onyekaba for help with the droplet freezing assays.

The investigation endeavored to study the distribution and allelic expressivity of hybrid necrosis genes (Ne1 and Ne2) in 332 maternal lines comprising 274 triticale × wheat derived homozygous recombinants, 41 winter × spring and spring × spring wheat derived doubled haploid lines, 12 reconstituted wheat lines and 5 released bread wheat cultivars of north-west Himalayan regions utilizing Himalayan genetic resources. Hybridization was executed at Palampur and Lahaul and Spiti, India by crossing maternal parents with two known tester parents C306 (NeNene2ne2) and HD2380 (ne1ne1NeNe) to generate F1 hybrids. The necrotic lines were categorized into Ne1 and Ne2-carriers based on the presence of necrosis genes in F1 hybrids. Subsequent screening revealed the presence of 83 Ne2-carriers (25%) and 249 non-carrier genotypes (75%) in the present study. Eighty-three Ne2-carriers comprised 51 triticale × wheat derived homozygous recombinants (11 Ne, 6 Ne, 26 Ne, 4 Ne and 4 Ne alleles), 22 winter × spring and spring × spring wheat derived doubled haploid lines (7 Ne, 12 Ne and 3 Ne alleles), 9 reconstituted wheat lines (5 Ne, 2 Ne and 2 Ne alleles) and 1 released variety (Ne allele) for necrosis and none of the lines possessed Ne1 gene. Analysis of Ne2 gene and their linkage properties with other paramount genes make Ne2-carriers superior source for emphasizing the genetic amelioration of wheat through selective hybridization.

The word Shvitra means in Sanskrit is Shvitra, which is white patch, without Kandu and inflammation, it present on anywhere of body, In this disease white spots or white discoloration takes place on the skin so it is named as Shvitra. Vitiligo is a long-term condition where pale white patches develop on the skin. It's caused by the lack of melanin, a pigment in the skin. In modern Science PUVA therapy and corticosteroid are used for treatment of disease but they may cause very harmful effect like skin burn, nausea dermatoheliosis that is ultraviolet ageing of skin. There is need Safe easier less complicate cost effective Ayurvedic treatment. Abhyanga stimulates Bhrajjak pitta which is present in the skin and helps restoring color. Jyotishamati Taila has extra medicinal property so it treats disease more comfortably.

Communicated by S.P. Baenziger) The present research endeavor was undertaken to depict the response of different generations viz., F1, F2, BC1F1, BC1F2, BC1F3, BC1F4 and BC1F5 of triticale × wheat and wheat × rye hybrids towards the different parameters of haploid induction. The experimental material included the different generations obtained utilizing five genotypes of triticale (DT-123, DT-126, TL-2900, TL-2908 and TL-9335), four genotypes of Himalayan rye (Karoki rye, shanoor rye, tino rye and triloki rye) with various elite bread wheat genotypes as parents in wide hybridization programme. The triticale × wheat and wheat × rye recombinants were further subjected to Imperata cylindrica-mediated chromosome elimination approach of doubled haploidy breeding. The variability in the haploid induction parameters was observed to be under genetic control for embryo formation and regeneration, while pseudoseed formation was only affected by auxin treatment. Among the different generations, the backcross generations viz., BC1F1 and BC1F2 were found to exhibit significant positive response towards haploid induction parameters in both triticale × wheat and wheat × rye hybridization. Knowledge of effective generation for haploid induction in triticale × wheat and wheat × rye hybridization not only saved the time and energy but also enhanced the efficiency of haploid induction.

An investigation to standardize the protocol for in vitro application of colchicine for enhancing the doubled haploid production in wheat was done. Two tetraploid (PDW-314 and A-9-30-1); and two hexaploid (DH-40 and C-306) wheat genotypes were used as maternal parents, whereas, the pollen sources involved Zea mays (cv. Bajaura Makka) and Imperata cylindrica. During the rabi seasons of years 2013–14 and 2014–15, wheat × maize and wheat × I. cylindrica hybridization was carried out followed by treatment of their haploids produced as a result of elimination of chromosomes of maize and I. cylindrica respectively, with varied doses of colchicine for different durations The various doses of colchicine were categorized into two groups: lower doses for longer durations (0.01, 0.025, 0.05% each for 5, 7, 9, 11 hrs) and higher doses for shorter durations (0.05, 0.075, 0.10, 0.15, 0.20, 0.25% each for 5, 4, 3, 2 hrs). The response of different concentrations of colchicine applied for varied durations revealed significant differences for various doubled haploidy parameters viz., per cent survived plants, per cent doubled haploid formation and per cent doubled haploid seed formation. In hexaploid and tetraploid wheats, colchicine doses of 0.075% for 4 hrs and 0.15% for 4 hrs, respectively were established as optimum for enhanced doubled haploid production.

Crop productivity in plants is reduced mostly because of a varied range of surrounding stresses, which are classified according to the nature of the stress, i.e., abiotic (non-living factors) stresses and biotic (living organisms) stresses. Abiotic stress factors include drought, salinity, extreme temperatures, water logging, heavy metals or minerals, etc. Traditional plant breeding systems and transgenic approaches have allowed breeders to produce improved crop varieties with agronomic qualitative/quantitative traits. The ever-evolving innovative advancements in molecular biology and biotechnology have immense potential for the future. Multifaceted reciprocity between a plant and a pathogen and the relevant stress response involves quite a few mechanisms responsible for the outbreak of a disease. To attain resistance against plant pathogens through clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9, significant knowledge about the genetics of resistance toward a particular disease and interrelated factors is important.

The COVID‐19 pandemic, caused by the SARS‐CoV‐2 coronavirus, has triggered a worldwide health emergency. Here, we show that ferritin‐like Dps from hyperthermophilic Sulfolobus islandicus, covalently coupled with SARS‐CoV‐2 antigens via the SpyCatcher system, forms stable multivalent dodecameric vaccine nanoparticles that remain intact even after lyophilisation. Immunisation experiments in mice demonstrated that the SARS‐CoV‐2 receptor binding domain (RBD) coupled to Dps (RBD‐S‐Dps) elicited a higher antibody titre and an enhanced neutralising antibody response compared to monomeric RBD. A single immunisation with RBD‐S‐Dps completely protected hACE2‐expressing mice from serious illness and led to viral clearance from the lungs upon SARS‐CoV‐2 infection. Our data highlight that multimerised SARS‐CoV‐2 subunit vaccines are a highly efficacious modality, particularly when combined with an ultra‐stable scaffold., Preparation and quality control of coupled antigen–Dps complexes (Ag‐S‐Dps). (A) SDS/PAGE of the three expressed and purified antigens as introduced in Fig. 1C, Coomassie stained. Glycosylation of Spike leads to a fuzzy appearance of its band. RBD‐SpyT2 and Spike‐SpyT2 were expressed in mammalian cells, and SpyT2‐NP was expressed in bacteria, as was the SpyC‐Dps scaffold. (B) Size exclusion chromatography to separate excess antigens after the SpyCatcher/Spytag2 coupling reactions; Superose 6 Increase in PBS. (C) SDS/PAGE of the coupled and purified Ag‐S‐Dps complexes. ‘RT’, no heating; ‘99’, heated to 99 °C. The SpyC‐Dps scaffold alone, as well as all the three coupled complexes show high‐molecular weight complexes, presumably dodecameric, that disappear only after heating of the samples in SDS loading buffer (Coomassie stained). (D) Negative‐stain electron microscopy analyses of the three multimeric Ag‐S‐Dps complexes, showing that all samples form defined and monodisperse spheres that display the antigens on their surface, leading to particles of different sizes for the three differently sized antigens.

The COVID-19 pandemic, caused by the SARS-CoV-2 coronavirus, has triggered a worldwide health emergency. So far, several different types of vaccines have shown strong efficacy. However, both the emergence of new SARS-CoV-2 variants and the need to vaccinate a large fraction of the world’s population necessitate the development of alternative vaccines, especially those that are simple and easy to store, transport and administer. Here, we showed that ferritin-like Dps protein from hyperthermophilic Sulfolobus islandicus can be covalently coupled with different SARS-CoV-2 antigens via the SpyCatcher system, to form extremely stable and defined multivalent dodecameric vaccine nanoparticles that remain intact even after lyophilisation. Immunisation experiments in mice demonstrated that the SARS-CoV-2 receptor binding domain (RBD) coupled to Dps (RBD-S-Dps) shows particular promise as it elicited a higher antibody titre and an enhanced neutralising antibody response compared to the monomeric RBD. Furthermore, we showed that a single immunisation with the multivalent RBD-S-Dps completely protected hACE2-expressing mice from serious illness and led to efficient viral clearance from the lungs upon SARS-CoV-2 infection. Our data highlight that multimerised SARS-CoV-2 subunit vaccines are a highly efficacious modality, particularly when combined with an ultra-stable scaffold.

Influenza A virus (IAV) and SARS-CoV-2 (COVID-19) cause pandemic infections where cytokine storm syndrome and lung inflammation lead to high mortality. Given the high social and economic cost of respiratory viruses, there is an urgent need to understand how the airways defend against virus infection. Here we use mice lacking the WD and linker domains of ATG16L1 to demonstrate that ATG16L1-dependent targeting of LC3 to single-membrane, non-autophagosome compartments – referred to as non-canonical autophagy – protects mice from lethal IAV infection. Mice with systemic loss of non-canonical autophagy are exquisitely sensitive to low-pathogenicity IAV where extensive viral replication throughout the lungs, coupled with cytokine amplification mediated by plasmacytoid dendritic cells, leads to fulminant pneumonia, lung inflammation and high mortality. IAV was controlled within epithelial barriers where non-canonical autophagy reduced IAV fusion with endosomes and activation of interferon signalling. Conditional mouse models and ex vivo analysis showed that protection against IAV infection of lung was independent of phagocytes and other leucocytes. This establishes non-canonical autophagy in airway epithelial cells as a novel innate defence that restricts IAV infection and lethal inflammation at respiratory surfaces., The EMBO Journal, 40 (6), ISSN:0261-4189, ISSN:1460-2075

The ability of acquired immune responses against SARS-CoV-2 to protect after subsequent exposure to emerging variants of concern (VOC) such as B1.1.7 and B1.351 is currently of high significance. Here, we use a hamster model of COVID-19 to show that prior infection with a strain representative of the original circulating lineage B of SARS-CoV-2 induces protection from clinical signs upon subsequent challenge with either B1.1.7 or B1.351 viruses, which recently emerged in the UK and South Africa, respectively. The results indicate that these emergent VOC may be unlikely to cause disease in individuals that are already immune due to prior infection, and this has positive implications for overall levels of infection and COVID-19 disease.

Magnaporthe oryzae that causes blast disease in rice is one of the most notorious phytopathogenic fungi responsible for severe economic loss in rice production. The fungus is one among the top ten devastating biotic threats to food security worldwide. To counter the pathogen and minimize the losses, various biological, conventional, and advanced molecular approaches have been utilized. Conventional and marker-assisted selection (MAS) breeding methods were deployed and varieties having tolerance or resistance have been developed. Durable and broad-spectrum resistance is the demand of the hour to protect the crop against blast disease, and several studies have made significant progress in this direction. Identification and molecular characterization of different blast resistance genes (R) and defense regulators (DR), and their deployment in various cultivars, is the most preferred approach to breed the disease-resistant varieties. The inclusive approaches including quantitative trait locus (QTL), association mapping, allele mining, bioinformatics tools and genetic engineering, miRNA, genome editing, CRISPR/Cas9 etc. have contributed to the strengthening of molecular breeding and providing of new opportunities for the development of rice varieties resistant to blast. Recent advances show that R-genes Pikh, Pi-1, Pi9, Pi20, Pi27, Pi39, Pi40, and Pita impart broad-spectrum resistance against blast disease. The bsr-d1, bsr-k1, spl11, spl33, and OsBBI1 transcription factors have also been reported for broad-spectrum resistance. The R-genes are broadly classified into eight groups, and in plants, the NBS-LRR genes correspond to the major group. These R-genes are the master keys of the entire defense system of plants, act in multilayered surveillance, and are governed by various extra- and intracellular receptor molecules to ward off pathogen invasion. The updated information on rice blast pathogen provided here will help to understand the different mechanism, advancement in resistance gene research, and focus on the breeding program in rice against blast disease.

Background Halophiles offer an attractive source of genes conferring salt tolerance. Halobacillus trueperi SS1 strain of Lunsu, Himachal Pradesh, India, a strict halophile, was exploited to isolate and clone the genes for salt tolerance. The genomic library of BamH1 digest of H. trueperi SS1 was constructed in pUC19, and recombinants were screened for salt tolerance on an LB medium containing ampicillin (100 μg/ml) and NaCl (0 to 1.5 M). Results One recombinant clone named as salt-tolerant clone (STC) conferred salt tolerance to host Escherichia coli/DH5α, which showed growth in the LB medium supplemented with ampicillin and 1.2 M NaCl. Restriction digestion and PCR analysis revealed the presence of an insert of approximately 2000 bp in the STC. DNA sequencing of the 2-kb insert on both strands yielded a sequence of 2301 nucleotides. Protein BLAST analysis of 2301-bp sequence of H. trueperi SS1 present in STC showed 97% identity to multidrug transport ATP binding/permease protein of Halobacillus karajensis. The insert contained in STC was subcloned into pGEX4T2 vector, and the recombinant clone STC/pGEX4T2 conferred salt tolerance to the bacterial host E. coli. Conclusions The present study led to the isolation of salt tolerance gene encoding a putative multidrug transport ATP binding/permease protein from H. trueperi SS1. The salt tolerance gene can be subcloned for transferring salt tolerance traits into agricultural crop plants for cultivation in saline and coastal lands.

Rare autoinflammatory diseases are a group of recurrent inflammatory genetic disorders caused due to genetic mutations in over 37 genes and typically have a recessive mode of inheritance. While a number of these disorders have been identified and reported from the Middle Eastern populations, the carrier frequency of these genetic mutations in the Middle Eastern populations is not known. The availability of whole-genome and exome datasets of over a thousand individuals from Qatar persuaded us to explore the genetic epidemiology of rare autoinflammatory genetic variants in the Middle East. We have systematically analyzed genetic variants in genome-scale datasets from Qatar with a compendium of variants associated with autoinflammatory diseases. The variants were further systematically reclassified according to the American College of Medical Genetics and Genomics (ACMG & AMP) guidelines for interpretation of pathogenicity of sequence variants. Our analysis identified 7 pathogenic and likely pathogenic variants with a significant difference in their allele frequencies compared to the global population. The cumulative carrier frequency of these variants was 2.58%. Furthermore, our analysis revealed that 5 genes implicated in rare autoinflammatory diseases were under natural selection. To our best knowledge, this is the first and comprehensive study on the population-scale analysis and genetic epidemiology for genetic variants causing rare autoinflammatory disease in any Middle Eastern population. Keywords: Autoinflammatory disease, Qatar, Middle East, genome, Epidemiology

Zika fever is a disease caused by Zika virus which is a mosquito-borne flavivirus and infection is through the bite of mosquito from the Aedes genus. Other mode of transmission of Zika fever is blood transfusion, which is thoroughly investigated. Prominent symptoms of Zika virus fever are mild fever, skin rash, conjunctivitis, muscle and joint pain, malaise or headache. These symptoms normally last for 2–7 days. There is currently no effective and easily affordable vaccine available against Zika virus. Keeping the current status in mind we have devised our objectives to design a peptide-based vaccine against Zika Virus using in-silico approach. The NS3 and NS5 proteins of Zika virus are well known of having important role in replication of the virus that is why we have targeted these proteins for epitope prediction. Prediction and screening of epitopes was performed using IEDB tool. We have also checked the efficacy of these screened epitopes with the help of IC50 value and docking score. “VEMGEAAAI” was found to be the high affinity binding epitope for MHC Class I and MHC Class II molecule. These findings conclude that the predicted epitope could well be used as a potential vaccine candidate for formulations of vaccines against Zika virus.

An efficient direct in vitro plant regeneration protocol for Saussurea costus (kuth) was developed using shoot tip explants derived from in vitro grown seedlings to generate genetically uniform plants. Murashige and Skoog (MS) medium supplemented with 1.14 µM thiadiazuron and 2.68 µM naphthalene acetic acid was found to be most optimal for highest for average shoot regeneration (73.33%) with maximum average number of shoots (11.4) and average shoot length (4.17 cm). For in vitro multiplication, shoots cultured on MS medium supplemented with 4.44 µM benzyl adenine, 2.32 µM kinetin and 1.44 µM gibberellic acid were found to grow best with 20.7 average shoot multiplication rate after 5–6 weeks. MS medium supplemented with 2.46 µM indole butyric acid was found to be the best medium for rooting of microshoots (77.78% rooting with 6.0 average number of roots and 3.07 cm average root length). The in vitro raised plantlets were successfully acclimatized under ex vitro conditions. The true to type nature of the in vitro raised plants was confirmed using DNA based ISSR markers for assessment of genetic stability of micropropagated plants. The standardized micropropagation protocol under present study may be tuned towards mass production of roots which are the most economical part of the plant. This would be useful as prerequisite for carrying out any genetic transformation studies while concentrating on any desirable trait in the same herb.

Ralstonia solanacearum phylotype II sequevar 1 (RsII-1, formerly race 3 biovar 2) causes tomato bacterial wilt, potato brown rot, and Southern wilt of geranium. Strains in RsII-1 cause wilting in potato and tomato at cooler temperatures than tropical lowland R. solanacearum strains. Although periodically introduced, RsII-1 has not established in the United States. This pathogen is of quarantine concern and listed as a Federal Select Agent. We report a rapidly sequenced (

Fluorescent Pseudomonas species have a number of traits like antifungal, siderophores, phosphate solubilization, lytic enzymes, HCN, ammonia and plant growth regulators that allow them to act as plant growth promoting and biocontrol agents. They competitively colonize plant roots, play important role in enhancing the growth of plants and in plant disease control. These may be used for soil improvement and to increase soil fertility hence for management of replant problem of apple in Himachal Pradesh. To ensure the sustained availability of PGP and biocontrol agent’s in soil formulation development protocol has to be standardized. On the basis of different PGPR activities three indigenous strains viz., Pseudomonas aeruginosa C, Pseudomonas fluorescens M and Pseudomonas putida L were selected for field studies. In all the treatments with individual (L, M and C) and consortia strains (L+M, L+C, M+C, L+M+C) there was an 8.0 to 86.5 per cent increase in plant height as compared to control. The performance of replanted apple was much better in terms of root colonization capacity, plant establishment and increase in plant growth in terms of plant height, number of nodes and branches over their respective control after fifteen months of plantation.

SummaryRespiratory viruses such as influenza A virus (IAV) and SARS-CoV-2 (Covid-19) cause pandemic infections where cytokine storm syndrome, lung inflammation and pneumonia lead to high mortality. Given the high social and economic cost of these viruses, there is an urgent need for a comprehensive understanding of how the airways defend against virus infection. Viruses entering cells by endocytosis are killed when delivered to lysosomes for degradation. Lysosome delivery is facilitated by non-canonical autophagy pathways that conjugate LC3 to endo-lysosome compartments to enhance lysosome fusion. Here we use mice lacking the WD and linker domains of ATG16L1 to demonstrate that non-canonical autophagy protects mice from lethal IAV infection of the airways. Mice with systemic loss of non-canonical autophagy are exquisitely sensitive to low-pathogenicity murine-adapted IAV where extensive viral replication throughout the lungs, coupled with cytokine amplification mediated by plasmacytoid dendritic cells, leads to fulminant pneumonia, lung inflammation and high mortality. IAV infection was controlled within epithelial barriers where non-canonical autophagy slowed fusion of IAV with endosomes and reduced activation of interferon signalling. This was consistent with conditional mouse models and ex vivo analysis showing that protection against IAV infection of lung was independent of phagocytes and other leukocytes. This establishes non-canonical autophagy pathways in airway epithelial cells as a novel innate defence mechanism that can restrict IAV infection and lethal inflammation at respiratory surfaces.

Spermatogenesis is a series of complex events involving a delicate balance between cell proliferation and cell differentiation. Aggregation of chromatins and epigenetic modifications play a vital role in spermatogenesis via regulation of molecular pathways to maintain testicular homeostasis. These epigenetic mechanisms consist of histone modification, chromatin remodelling, DNA methylation and miRNA, etc., which reportedly are critical players in spermatogenesis. One such mechanism involves regulation of oxidative stress in the male reproductive system. The fact that testicular cells contain plenty of unsaturated fatty acids and undergo division at a high rate makes spermatogenic cells highly susceptible to oxidative insult leading to deleterious effect on spermatozoa, which may culminate in infertility in men. Although the correlation between ROS-mediated oxidative stress and epigenetic alterations has been indicated, research in this regard is still in infancy. Further, the fact that environmental and life style factors are critical determinants of spermatogenic potential indicates the importance of epigenetic regulation of key molecular events in spermatogenesis. Therefore, the current review aims to discuss the ROS-induced epigenetic deregulation of the molecular mechanism(s) involved in spermatogenesis.

Background Thermostable cellulases are in constant demand for several biotechnological applications. Two thermophilic bacterial strains PW1 and PW2 isolated from Tattapani hot spring were found to have cellulolytic activity. Subsequently, PW1 and PW2 were identified and mined for genes encoding cellulase activity. Results Sequencing of the 16S rDNA of PW1 and PW2 identified them as Bacillus sp. PW1 (Acc no. KU711837) and Bacillus sp. PW2 (Acc no. KU711838), respectively, which clustered in the clades containing thermophilic members of Bacillus sp. and Geobacillus species. Phylogenetic analysis revealed that despite the morphological and sequence identities, Bacillus sp. PW1 and Bacillus sp. PW2 are different at the genetic level. The cellulase genes (~ 1.1 kb) of the two bacterial strains were amplified using primers designed against related thermophilic cellulases. Sequencing of the cellulase gene amplicons of PW1 and PW2 revealed that they encode proteins of 280 and 206 amino acid residues, respectively. Sequence and domain analysis of the protein products of PW1 and PW2 revealed that they belong to M42 family of aminopeptidase/endoglucanase. The PW2 endoglucanase coding sequence was submitted to Genbank under accession no. MH049504. The structures of putative endoglucanases of PW1 and PW2 were generated using 1VHE.A as template, which showed the presence of vast proportion of random coils. Molecular docking of the modeled endoglucanase proteins with various substrates and products of cellulases showed that carboxymethyl cellulose and maltose exhibit the highest binding affinity, while xylan and glucose the least. Conclusions The two thermophilic bacteria PW1 and PW2 and their endoglucanase gene can be further utilized for recombinant production of thermostable cellulases for their application in industries.

Routine strain-level identification of plant pathogens directly from symptomatic tissue could significantly improve plant disease control and prevention. Here we tested the Oxford Nanopore Technologies (ONT) MinION™ sequencer for metagenomic sequencing of tomato plants either artificially inoculated with a known strain of the bacterial speck pathogenPseudomonas syringaepv.tomato(Pto), or collected in the field and showing bacterial spot symptoms caused by either one of fourXanthomonasspecies. After species-level identification using ONT’s WIMP software and the third party tools Sourmash and MetaMaps, we used Sourmash and MetaMaps with a custom database of representative genomes of bacterial tomato pathogens to attempt strain-level identification. In parallel, each metagenome was assembled and the longest contigs were used as query with the genome-based microbial identification Web service LINbase. Both the read-based and assembly-based approaches correctly identifiedPtostrain T1 in the artificially inoculated samples. The pathogen strain in most field samples was identified as a member ofXanthomonas perforansgroup 2. This result was confirmed by whole genome sequencing of colonies isolated from one of the samples. Although in our case, metagenome-based pathogen identification at the strain-level was achieved, caution still needs to be exerted when interpreting strain-level results because of the challenges inherent to assigning reads to specific strains and the error rate of nanopore sequencing.

Soil and water pollution with heavy metals is a worldwide issue and is still unresolved. Phytoremediation is a new and rising technology that applies plants for cleanup lightly contaminated soils and water. Lasiurus scindicus have been chosen in this work for decontaminating soil and water from heavy metal (lead). This plant was grown in contaminated soil and water over a 105 day pot trial period. Accruement of the heavy metal was studied after 45, 65, 85 and 105 days in leaves and roots by AAS. It was observed that high accumulation level of lead in root after 45 and 65 days. The results conclude that the Lasiurus scindicus has phytoremediation potentiality and could be used in restoring soil polluted with Lead.

A rationale use of the genetic resources in breeding programs requires an understanding of relationships within and among related crop varieties. In the present study, genetic relatedness among fifty genotypes of temperate pome fruit crops (apple, pear, quince and loquat) of family Rosaceae growing in Himachal Pradesh (India) was tested using arbitrary oligonucleotide random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) molecular marker analyses. The level of polymorphism across the subjected genotypes was 97.29 and 97.45 % by RAPD and ISSR markers, respectively. The dendrograms derived from RAPD and ISSR analysis grouped temperate pome fruit crops into two major clusters comprising of 40 and 10 genotypes which branched at similarity value of 0.31 and 0.33, respectively. Estimates of pooled similarity coefficient ranging from 0.30 to 0.94 indicated their promise for application to future molecular screening, map construction and comparative genomic studies, etc. Therefore, these results could be helpful in framing more extensive studies on the determined taxa.

The Arab population encompasses over 420 million people characterized by genetic admixture and a consequent rich genetic diversity. A number of genetic diseases have been reported for the first time from the population. Additionally a high prevalence of some genetic diseases including autosomal recessive disorders such as hemoglobinopathies and familial mediterranean fever have been found in the population and across the region. There is a paucity of databases cataloguing genetic variants of clinical relevance from the population. The availability of such a catalog could have implications in precise diagnosis, genetic epidemiology and prevention of disease. To fill in the gap, we have compiled DALIA, a comprehensive compendium of genetic variants reported in literature and implicated in genetic diseases reported from the Arab population. The database aims to act as an effective resource for population-scale and sub-population specific variant analyses, enabling a ready reference aiding clinical interpretation of genetic variants, genetic epidemiology, as well as facilitating rapid screening and a quick reference for evaluating evidence on genetic diseases.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). Sequencing the viral genome as the outbreak progresses is important, particularly in the identification of emerging isolates with different pathogenic potential and to identify whether nucleotide changes in the genome will impair clinical diagnostic tools such as real-time PCR assays. Although single nucleotide polymorphisms and point mutations occur during the replication of coronaviruses, one of the biggest drivers in genetic change is recombination. This can manifest itself in insertions and/or deletions in the viral genome. Therefore, sequencing strategies that underpin molecular epidemiology and inform virus biology in patients should take these factors into account. A long amplicon/read length-based RT-PCR sequencing approach focused on the Oxford Nanopore MinION/GridION platforms was developed to identify and sequence the SARS-CoV-2 genome in samples from patients with or suspected of COVID-19. The protocol, termed Rapid Sequencing Long Amplicons (RSLAs) used random primers to generate cDNA from RNA purified from a sample from a patient, followed by single or multiplex PCRs to generate longer amplicons of the viral genome. The base protocol was used to identify SARS-CoV-2 in a variety of clinical samples and proved sensitive in identifying viral RNA in samples from patients that had been declared negative using other nucleic acid-based assays (false negative). Sequencing the amplicons revealed that a number of patients had a proportion of viral genomes with deletions.

We report here the genome sequence of halophilic Halobacillus trueperi SS1, isolated from the Lunsu saltwater body in India. The bacteria are Gram positive and rod shaped., We report here the genome sequence of halophilic Halobacillus trueperi SS1, isolated from the Lunsu saltwater body in India. The bacteria are Gram positive and rod shaped. The genome of H. trueperi SS1 has 4.14 Mbp, with 4,329 coding sequences, 35 RNA genes (29 tRNAs, 2 rRNAs, and 4 noncoding RNAs), and 42.15% G+C content.

Biological control agents (BCAs) are gaining more attention as alternatives to the chemical pesticides for management of pests and diseases in agricultures. It offers an ecofriendly way for plant health management (PHM), and it also helps to reduce the excessive use of toxic chemicals or pesticides. The BCAs considered as the most promising technology for sustainable agriculture. Various beneficial BCAs have been known for PHM but need effective adoption together with standardization of bioformulations for field applications. The mechanisms of BCAs in PHM mainly include direct (parasitism, hyperparasitism, commensalism), indirect (competition, systemic acquired or induced systemic resistance), or mixed antagonistic modes (production of antibiotics, siderophores, lytic enzyme, and volatile organic substances). Apart from their role in plant growth promotion, plant growth-promoting rhizobacteria (PGPR) and fungi (PGPF) also act as BCAs that function through either of the mechanisms. The PGPR/PGPF possesses many traits that serve them as potential BCAs, and therefore possess great promise for successful application in sustainable agriculture. In this chapter, different approaches for BCAs applications and their mechanism in PHM especially for disease and pest (DP) management are discussed. Also, the relationship between the PGPR/PGPF diversity and their application in biological control is discussed. Furthermore, we have summarized the global-Indian scenario of current research including state-of-the-art technological advances in PGPR/PGPF-mediated DP management.

Here, we report the genome sequence of hyperthermophilic and halophilic Parageobacillus toebii PW12, isolated from the Tattapani hot spring in the northwest Himalayas. The genome size of Parageobacillus toebii PW12 is 3,210,377 bp., Here, we report the genome sequence of hyperthermophilic and halophilic Parageobacillus toebii PW12, isolated from the Tattapani hot spring in the northwest Himalayas. The genome size of Parageobacillus toebii PW12 is 3,210,377 bp. The G+C content is 42.05%, and 3,382 coding sequences (CDS), 80 tRNAs, 5 noncoding RNAs (ncRNAs), and 4 CRISPR arrays were predicted.

Summary Escherichia coli biofilm consists of a bacterial colony embedded in a matrix of extracellular polymeric substances (EPS) which protects the microbes from adverse environmental conditions and results in infection. Besides being the major causative agent for recurrent urinary tract infections, E. coli biofilm is also responsible for indwelling medical device-related infectivity. The cell-to-cell communication within the biofilm occurs due to quorum sensors that can modulate the key biochemical players enabling the bacteria to proliferate and intensify the resultant infections. The diversity in structural components of biofilm gets compounded due to the development of antibiotic resistance, hampering its eradication. Conventionally used antimicrobial agents have a restricted range of cellular targets and limited efficacy on biofilms. This emphasizes the need to explore the alternate therapeuticals like anti-adhesion compounds, phytochemicals, nanomaterials for effective drug delivery to restrict the growth of biofilm. The current review focuses on various aspects of E. coli biofilm development and the possible therapeutic approaches for prevention and treatment of biofilm-related infections.

A collection of 50 rhizospheric and 43 endorhizospheric bacteria was obtained from Podophyllum hexandrum grown in high hills of Churdhar, Himachal Pradesh, India and subjected to replica plating for one step screening of plant growth promoting (PGP) traits. After replica plating, 41 bacterial isolates simultaneously growing on Pikovskaya’s, Chrome-azurol-S, and Jensen’s medium were characterized for additional PGP traits. Of the total, 80.48 % isolates produced indole-3-acetic acid, 12.20 % produced HCN, 53.65 % exhibited chitinase and 41.46 % exhibited protease activity. In vitro inhibition of fungal pathogens viz., Alternaria solani, Fusarium oxysporum, Dematophora necatrix, Sclerotium rolfsii and Phytophthora spp. ranged from 44 % to 88 %. Twenty most efficient PGPR under in vitro conditions were evaluated for growth promotion of tomato in growth chamber. Significantly highest root length (43.00 mm), shoot length (73.00 mm), plant dry weight (12.90 mg plant−1) and vigour index (10,567.60) were recorded for isolate 2a1. The efficacy of isolate 2a1 was further evaluated under net house and resulted in remarkable increase in root length (58.02 %), shoot length (66.89 %), root dry weight (98.67 %), shoot dry weight (88.58 %), available nitrogen (31.89 %), phosphorus (22.15 %) and potassium (4.51 %) content in soil over untreated control. Remarkable increase was also recorded in nitrogen (11.85 %), phosphorus (23.08 %) and potassium (36.84 %) content of shoot biomass over control. Strain 2a1 was identified as Bacillus subtilis after biochemical and phylogenetic analysis. These results demonstrated that strain 2a1 from Podophyllum hexandrum exhibited multiple PGP attributes for development as a biofertilizer to enhance soil fertility and promote plant growth.

Background: Management of furcation defects is challenging, and constantly newer therapeutic strategies are evolving. Platelet-rich fibrin (PRF) is successfully used alone and in combination with various agents in the furcation defects. Lately, metformin (MF), a second generation biguanide has gained popularity owing to its osteogenic potential. Aims and Objectives: The aim of the present study was to evaluate the clinical and radiographic effectiveness of open flap debridement (OFD) and PRF when compared to OFD + PRF + 1% MF gel in the management of mandibular Grade II furcation defects. Materials and Methods: Thirty mandibular grade II furcation defects were stratified into two groups; in one group OFD and PRF is used, and the other group had an additional MF gel with PRF in OFD. Clinical parameters such as plaque index, modified sulcus bleeding index, probing pocket depth (PD), relative vertical attachment level (RVAL), and relative horizontal attachment level (RHAL) were recorded at baseline and at 6 months. Radiovisiography and ImageJ software were used to evaluate the intrabony defect depth. Results: The OFD + PRF + MF group showed significantly higher probing PD reduction, RVAL and RHAL gain than the OFD + PRF group. Conclusions: PRF when combined with a potential osteogenic agent like MF can provide a better therapeutic benefit to a furcation involved tooth.

Summary Summary In the present study, production, characterization and optimization of bacterial cellulose (BC) from Gluconacetobacter xylinus MTCC 7795 has been investigated. Effect of various carbon sources and bioprocess parameters on bacterial growth rate and subsequent BC production were studied and optimized. Supplementation of the media with different carbon sources resulted in varying yield of bacterial cellulose. Variations in the arrangement of bacterial cellulose with variations in carbon source were analyzed using FT-IR. Results have suggested that characteristic vibrational modes of BC produced were almost same in the typical fingerprint regions as in commercial cellulose.

The importance of germplasm characterization is an important link between the conservation and utilization of plant genetic resources in various breeding programmes. In the present study, genetic variability and relationships among twenty-four Capsicum annuum L. genotypes were tested using PCR-based RAPD and ISSR molecular markers. The level of polymorphism across genotypes was 42–44 % as revealed by RAPD and ISSR, respectively. The highest similarity was detected between PC-1, Feroz, Gajio, Kandaghat Selection-9 genotypes (96 %) and Feroz, Kandaghat Selection-9 (93 %), by RAPD and ISSR markers, respectively. It was concluded from the pooled analysis of both molecular markers that genotypes ACC-2 and Mahog were most distantly related to each other. Hence, it is recommended that these two genotypes should be crossed to create a segregating population with maximum genetic diversity. Further, it is suggested that molecular markers are valid tags for the assessment of genetic diversity in C. annuum germplasm.

Diabetes is a chronic progressive endocrine disease characterized by elevation of blood glucose level. This disease is associated with decreased life expectancy; increased morbidity and diminished quality of life. Measuring QOL is important, because by doing so we can predict the patient's capacity as well as ability to manage the disease and to maintain overall well-being and the long-term health. The objective of the study was to study the correlation of glycaemic control, age and duration of disease with quality of life in patients with Type 2 Diabetes Mellitus.

Substance abuse has become one of the major contributor to psychiatric morbidity worldwide. The pattern of substance abuse, in patients attending psychiatry OPD of IGMC, Shimla, from the years 2006 to 2008, was studied. Alcohol abuse was found to be the most prevalent, followed by that of cannabis. Opioid, sedative and inhalant abuse trends were found increase over these years. One of the objectives of the United Nations International Drug Control Programme is to improve the quality and comparability of country - specific drug abuse data by developing an information infrastructure in some 100 countries. The Annual Reports Questionnaire (ARQ) is used to gather information from various international and regional organizations, research institutes, and epidemiology networks. Based on the most recent data collected, it is estimated that:  Around 8 million people abuse opiates / heroin, mostly in South East and South West Asia. Heroin abuse is far more wide-spread than it was a decade ago. Sharp increases have been reported in former Soviet Union countries. Increases also have been reported in Iran, Turkey, and Pakistan. Heroin has remained readily available in major metropolitan areas in the United States.  Some 13 million people abuse cocaine worldwide, with the highest prevalence reported in the United States. Many Latin American countries also have high levels of cocaine and bazuco (coca paste) abuse. In the 1995 to 1997 period, cocaine abuse spread in Europe and was stabilized in the main markets of North America. Increases have been reported in the Andean and neighboring countries and along trafficking routes to America and Europe.  141 million people consume cannabis annually. It is the most widely abused drug in all parts of the world. Cannabis abuse is particularly high in western Africa, Oceania, Central America, North America, and a number of European countries. 2