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Musa sp. cultivar Rasthali (Silk AAB) is a choice variety of the Asian sub-continent. Its production and sustenance are threatened by Fusarium wilt, which affects the livelihoods of small and marginal farmers. The use of quality planting material is one of the strategies to manage the disease. Availability of quality planting material for varieties other than Grand Naine is limited. Large-scale micropropagation using existing technologies is laborious and expensive. Temporary immersion bioreactor system is emerging as a potential advancement in the micropropagation industry. In this study, a cost-effective temporary immersion bioreactor (TIB) system has been developed and an efficient micropropagation method has been standardized. Explants cultured in TIB with 250 ml of culture medium in a 2-min immersion frequency of 6 h were found to be efficient for shoot proliferation and rooting. Its efficacy has been compared with the semisolid culture method. At the end of the 6th subculture, 1496 ± 110 shoots per explant were obtained in TIB. Chlorophyll, carotenoid, stomatal index, and the number of closed stomata were examined to determine the physiological functions of the plants grown in TIB and compared with semisolid grown plantlets. Plantlets grown in TIB were genetically stable and were confirmed using inter-simple sequence repeat (ISSR) markers. The multiplication of shoots in TIB was 2.7-fold higher than the semisolid culture method, which is suitable for large-scale production of planting material for commercial applications.

The presence of residual female fertility in most of the parthenocarpic banana accessions encourages the banana breeder to develop new hybrids through conventional breeding. Desirable trait can be fixed in the first generation of hybrid progenies, but the evaluation of these hybrids in field is the time-consuming process owing to non-availability of uniform suckers/planting material. This can be overcome by developing multiple shoots from single embryo in a short period of time through embryo culture. A protocol for in vitro multiplication of plantlets from zygotic embryos was standardized in seeded accessions. Multiple shoots from zygotic embryos were achieved up to 55.2% and 64.1% in seeded accessions of Musa acuminata and M. velutina respectively in medium supplemented with 17.76 µM of BAP. The Single shoot derived (only germination) from zygotic embryos was decapitated and the apical meristem were disturbed for further multiple shoot formation in media supplemented with 17.76 µM of BAP. Present studies revealed that in total 75% and 91% of the M. acuminata and M.velutina embryos were able to produce multiple shoot from single embryo by manipulating the media composition and decortications technique. The above protocol was applied for zygotic embryos obtained from controlled pollination (18 cross combinations) and open pollination (nine accessions) of various genomic groups (ABB, AAB, AA). The multiple shoots derived from zygotic embryos and plantlet germinated from zygotic embryos was examined for genetic fidelity analysis by SSR markers. The protocol for multiple shoot formation from single zygotic embryo under in vitro culture developed in this study will accelerate the banana breeding program.

Somatic embryogenesis is an important tool for crop improvement through transgenic approach and even for gene editing. It has been hypothesized regularly for large-scale propagation of banana which necessitates basic data on genetic fidelity and field performance of the plants towards ensure the commercial feasibility of the technique. Plantlets regenerated from embryogenic cell suspension (ECS) cultures established using immature male flower buds were examined for genetic fidelity using Inter Simple Sequence Repeats (ISSR) markers. Results showed that the primers UBC 808, UBC 811 and UBC 841 each generated one polymorphic band with an overall variation in banding pattern of 3.34 and 2.09% in cvs. Grand Naine and Rasthali respectively. Field evaluation of the ECS derived plants showed that there were no negative effects on the vegetative and yield parameters. Remarkably no phenotypic off-types were observed in this field trial. The level of genetic variation observed in this study is not an obstacle for further uptake of this novel propagation technique. Field performance of ECS derived plants being on par with shoot tip cultured plants concludes that somatic embryogenesis could be successfully employed for commercial propagation of banana plantlets.

Cancer is a highly malignant disease, killing approximately 10 million people worldwide in 2020. Cancer patient survival substantially relies on early diagnosis. In this study, we evaluated whether genes involved in glucose metabolism could be used as potential diagnostic markers for cancer. In total, 127 genes were examined for their gene expression levels and pairwise gene correlations. Genes ADH1B and PDHA2 were differentially expressed in most of the 12 types of cancer and five pairs of genes exhibited consistent correlation changes (from strong correlations in normal controls to weak correlations in cancer patients) across all types of cancer. Thus, the two differentially expressed genes and five gene pairs could be potential diagnostic markers for cancer. Further preclinical and clinical studies are warranted to prove whether these genes and/or gene pairs would indeed aid in early diagnosis of cancer.

Splitting of the genus Penaeus sensu lato into six new genera based on morphological features alone has been controversial in penaeid shrimp taxonomy. Several studies focused on building phylogenetic relations among the genera of Penaeus sensu lato. However, they lack in utilizing full mitochondrial DNA genome of shrimp representing all the six controversial genera. For the first time, the present study targeted the testing of all the six genera of Penaeus sensu lato for phylogenetic relations utilizing complete mitochondrial genome sequence. In addition, the study reports for the first time about the complete mitochondrial DNA genome sequence of Fenneropenaeus indicus, an important candidate species in aquaculture and fisheries, and utilized it for phylogenomics. The maximum likelihood and Bayesian approaches were deployed to generate and comprehend the phylogenetic relationship among the shrimp in the suborder, Dendrobranchiata. The phylogenetic relations established with limited taxon sampling considered in the study pointed to the monophyly of Penaeus sensu lato and suggested collapsing of the new genera to a single genus. Further, trends in mitogenome‐wide estimates of average amino acid identity in the order Decapoda and the genus Penaeus sensu lato supported restoration of the old genus, Penaeus, rather promoting the creation of new genera., The maximum likelihood and Bayesian phylogenetic inferences based on complete mitochondrial DNA genome sequences supported monophyly of the genus, Penaeus sensu lato. The mitogenome‐based similarity metrics estimated for between‐genera within a family and also for between‐species within a genus in the order Decapoda corroborated phylogenetic inferences drawn for Penaeus sensu lato.

Shrimp aquaculture over the years has made tremendous progress. The application of modern biotechnological tools, including next-generation sequencing (NGS) techniques, has revolutionized the genom...

Objectives The search for new biocontrol agents, especially from natural sources, to control plant pathogens is a key area in tea plant protection research. Methods Starch casein nitrate (SCN) agar was used to isolate the actinomycetes from the soil samples. These isolates were tested with modified version of the dual-culture method for antagonistic activity against Pestalotiopsis theae. The most bioactive isolate of actinomycets has been identified by biochemical, phisological, and morphological characterisation. Bioactivity of Streptomyces sp. SLR03 metabolites was measured in vitro and in vivo. The extract was eventually analyzed by GC–MS. Results For the first time, an attempt has been made to isolate actinomycete species with bio-control potential from the river soil samples. A total of one hundred and seven actinomycete strains isolated and were evaluated for antagonistic potential against Pestalotiopsis theae using a dual-culture assay. Among the strains isolated, one strain SLR03 that showed potential and it was characterized. Further, the strain SLR03 was evaluated for antagonistic activity against P. theae both in vitro and in vivo. In in vitro assay, 86.15% and 93.85% mycelial growth inhibition was observed with cell-free filtrate and ethyl acetate extract of Streptomyces sp. SLR03, respectively. The ethyl acetate extract was further evaluated for its biocontrol activity against P. theae, it exhibited 80.39% reduction of disease incidence compared to the control. Further, the ethyl acetate extract were analyzed using GC–MS. The GCMS chromatogram exhibited 24 intense peaks consistently with 19 different compounds, 10 of which contain antifungal activity. Conclusions This present study illustrates that Streptomyces sp. strain SLR03 is a prospective candidature for forthcoming biological control programme.

The World Health Organization has declared COVID-19 as a global health emergency. COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and highlights an urgent need for therapeutics. Here, we have employed a series of computer-aided drug repurposing campaign to discover inhibitors of RNA dependent RNA polymerase (RdRp) and Nsp15/EndoU. Subsequently, MD simulation has been performed to observe dynamic behavior of identified leads at the active site of RdRp and Nsp15. We successfully identified novel lead molecule such as Alectinib for RdRp while Naldemedine and Ergotamine for NSP15. These lead molecules were accommodated in the active site of the enzyme and stabilized by the networks of the hydrogen bond, pi type and hydrophobic interaction with key residues of either target. Interestingly, identified compounds show molecular mimicry in terms of molecular interactions with key residues of RdRp and Nsp15 essential for catalysis and substrate interaction. Previously, Alectinib, Naldemedine and Ergotamine were used as drug in different diseases might be repurposed against selected protein targets of COVID19. Finally, we propose that the identified inhibitors represent a novel lead molecule to design a more effective inhibitor to stop the progress of pathogen. Communicated by Ramaswamy H. Sarma

Purpose The aquaculture sector is a major contributor to the economic and nutritional security for a number of countries. India’s total seafood exports for the year 2017–2018 accounted for US$ Million 7082. One of the major setbacks in this sector is the frequent outbreaks of diseases often due to bacterial pathogens. Vibriosis is one of the major diseases caused by bacteria of Vibrio spp., causing significant economic loss to the aquaculture sector. The objective of this study was to understand the genetic composition of Vibrio spp. Methods Thirty-five complete genomes were downloaded from GenBank comprising seven vibrio species, namely, Vibrio alginolyticus, V. anguillarum, V. campbellii, V. harveyi, V. furnissii, V. parahaemolyticus, and V. vulnificus. Pan-genome analysis was carried out with coding sequences (CDS) generated from all the Vibrio genomes. In addition, genomes were mined for genes coding for toxin-antitoxin systems, antibiotic resistance, genomic islands, and virulence factors. Results Results revealed an open pan-genome comprising of 2004 core, 8249 accessory, and 6780 unique genes. Downstream analysis of genomes and the identified unique genes resulted in 312 antibiotic resistance genes, 430 genes coding for toxin and antitoxin systems along with 4802, and 4825 putative virulent genes from genomic island regions and unique gene sets, respectively. Conclusion Pan-genome and other downstream analytical procedures followed in this study have the potential to predict strain-specific genes and their association with habitat and pathogenicity.

Bovine respiratory disease (BRD) and bovine mastitis (BM) are the most common and costly infectious diseases in beef cattle and dairy cattle, respectively. In the current study, we evaluated the antimicrobial activity of seven phytochemicals against twelve BRD- and/or BMcausing bacterial pathogens. Our results show that allyl isothiocyanate, benzyl isothiocynate, cinnamaldehyde and eugenol are effective against most of the BRD- and/or BM-causing bacterial pathogens and could be repurposed as alternatives to antibiotics for the prevention/elimination of BRD and BM in feedlots.

Shrimp production in India has gradually increased since 2009 with the introduction of Penaeus vannamei . Most coastal states in India have been important contributors to P. vannamei production. However, among the many challenges faced in sustainable shrimp farming, prevention of viral diseases and pond water quality management remain major concerns. In this study, 27 shrimp ponds located in Kalpakkam and Elavur regions of Tamil Nadu were monitored to characterise the pond water parameters including pH, salinity, temperature, alkalinity, ammonia, nitrite, hardness, dissolved oxygen (DO) and Vibrio spp. load. Sudden outbreaks of white spot syndrome virus (WSSV) were observed in some ponds which were found to be associated with variations in abiotic parameters. Parameters such as pH, DO and nitrite were observed to be within the permissible range, while temperature, salinity, Vibrio spp. load and ammonia were relatively higher in certain ponds. The influence of these abiotic factors triggering WSSV outbreak were investigated in this study. Keywords : Aquaculture, Penaeus vannamei, Vibrio spp., Water parameters, White spot syndrome virus

Green chemistry is considered as an ideal approach for the future prospect of nanomaterials. 70% of the earth’s surface is covered by a marine environment of which most of the area is still unexplored. In green chemistry, marine plants/algae/bio-nanofactories are utilized to orchestrate nanoparticles (NPs)/bimetallic NPs for biomedical and other applications. Algal extract (microalgae and macroalgae) intrinsically contains biomolecules, which leads to shell formation around the metal NPs to improve their biocompatibility. Different groups of algae such as phaeophyceae, rhodophyceae, chlorophyceae, and cyanophyceae have been explored for the amalgamation of metallic NPs. It is the ability of essential phytochemicals in algae (alkaloids, flavonoids, and terpenoids) which accumulate metal and reduce metal ions, thus making them superior contenders for synthesis by acting as sculpturing or capping agents, which directs particle growth. The effective biomolecules present in the algae influence the size, shape, composition, and physicochemical properties of NPs. Moreover, algae can grow even in polluted water sources and can synthesize NPs in these conditions without competing with the arable lands and freshwater bodies, thereby increasing their applicability to a wider extent. Holistically, this chapter will emphasize the comprehensive studies of algae-based nanoparticle synthesis in terms of their antimicrobial (antibacterial and antifungal) and anticancer activities.

Tryptophan metabolism plays essential roles in both immunomodulation and cancer development. Indoleamine 2,3-dioxygenase, a rate-limiting enzyme in the metabolic pathway, is overexpressed in different types of cancer. To get a better understanding of the involvement of tryptophan metabolism in cancer development, we evaluated the expression and pairwise correlation of 62 genes in the metabolic pathway across 12 types of cancer. Only gene AOX1, encoding aldehyde oxidase 1, was ubiquitously downregulated, Furthermore, we observed that the 62 genes were widely and strongly correlated in normal controls, however, the gene pair correlations were significantly lost in tumor patients for all 12 types of cancer. This implicated that gene pair correlation coefficients of the tryptophan metabolic pathway could be applied as a prognostic and/or diagnostic biomarker for cancer.

The Arp2/3 complex is essential for the assembly of branched filamentous actin, but its role in physiology and development is surprisingly little understood. Melanoblasts deriving from the neural crest migrate along the developing embryo and traverse the dermis to reach the epidermis, colonising the skin and eventually homing within the hair follicles. We have previously established that Rac1 and Cdc42 direct melanoblast migration in vivo. We hypothesised that the Arp2/3 complex might be the main downstream effector of these small GTPases. Arp3 depletion in the melanocyte lineage results in severe pigmentation defects in dorsal and ventral regions of the mouse skin. Arp3 null melanoblasts demonstrate proliferation and migration defects and fail to elongate as their wild-type counterparts. Conditional deletion of Arp3 in primary melanocytes causes improper proliferation, spreading, migration and adhesion to extracellular matrix. Collectively, our results suggest that the Arp2/3 complex is absolutely indispensable in the melanocyte lineage in mouse development, and indicate a significant role in developmental processes that require tight regulation of actin-mediated motility., Summary: This study demonstrates that Arp2/3-dependent branched actin assembly is indispensable for melanoblasts populating the skin of the developing mouse embryo.

The larvicidal potential of crude leaf extracts of Rhizophora mucronata, the red mangrove, using diverse solvent extracts of the plant against the early fourth instar larvae of Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti mosquito vectors was analyzed. The acetone extract of R. mucronata showed the greatest efficacy: for Cx. quinquefasciatus (LC50 = 0.13 mg/mL, LC90 = 2.84 mg/mL), An. stephensi (LC50 = 0.34 mg/mL, LC90 = 6.03 mg/mL), and Ae. aegypti (LC50 = 0.11 mg/mL, LC90 = 1.35 mg/mL). The acetone extract was further fractionated into four fractions and tested for its larvicidal activity. Fraction 3 showed stronger larvicidal activity against all the three mosquito larvae. Chemical characterization of the acetone extract displayed the existence of several identifiable compounds like phytol, 3,7,11,15-tetramethyl-2-hexadecen-1-ol, 1-hexyl-2-nitrocyclohexane, eicosanoic acid etc. Enzyme assay displayed that R. mucronata active F3-fractions exert divergent effects on all three mosquitos&rsquo, biochemical defensive mechanisms. The plant fractions displayed significant repellent activity against all the three mosquito vectors up to the maximum repellent time of 210 min. Thus, the bioactive molecules in the acetone extract of R. murconata leaves showed significant larvicidal and enzyme inhibitory activity and displayed novel eco-friendly tool for mosquito control.

Cells migrating over 2D substrates are required to polymerise actin at the leading edge to form lamellipodia protrusions and nascent adhesions to anchor the protrusion to the substrate. The major actin nucleator in lamellipodia formation is the Arp2/3 complex, which is activated by the WAVE regulatory complex (WRC). Using inducible Nckap1 floxed mouse embryonic fibroblasts (MEFs), we confirm that the WRC is required for lamellipodia formation, and importantly, for generating the retrograde flow of actin from the leading cell edge. The loss of NCKAP1 also affects cell spreading and focal adhesion dynamics. In the absence of lamellipodium, cells can become elongated and move with a single thin pseudopod, which appears devoid of N-WASP. This phenotype was more prevalent on collagen than fibronectin, where we observed an increase in migratory speed. Thus, 2D cell migration on collagen is less dependent on branched actin.

Summary Efficient motility requires polarized cells, with pseudopods at the front and a retracting rear. Polarization is maintained by restricting the pseudopod catalyst, active Rac, to the front. Here, we show that the actin nucleation-promoting factor Wiskott-Aldrich syndrome protein (WASP) contributes to maintenance of front-rear polarity by controlling localization and cellular levels of active Rac. Dictyostelium cells lacking WASP inappropriately activate Rac at the rear, which affects their polarity and speed. WASP’s Cdc42 and Rac interacting binding (“CRIB”) motif has been thought to be essential for its activation. However, we show that the CRIB motif’s biological role is unexpectedly complex. WASP CRIB mutants are no longer able to restrict Rac activity to the front, and cannot generate new pseudopods when SCAR/WAVE is absent. Overall levels of Rac activity also increase when WASP is unable to bind to Rac. However, WASP without a functional CRIB domain localizes normally at clathrin pits during endocytosis, and activates Arp2/3 complex. Similarly, chemical inhibition of Rac does not affect WASP localization or activation at sites of endocytosis. Thus, the interaction between small GTPases and WASP is more complex than previously thought—Rac regulates a subset of WASP functions, but WASP reciprocally restricts active Rac through its CRIB motif., Graphical Abstract, Highlights • WASP exploits its CRIB motif to remove active Rac from the membrane via endocytosis • WASP is recruited to clathrin-coated pits (CCPs) independently of small GTPases • WASP triggers actin polymerization at CCPs independently of small GTPase activation • WASP maintains homeostasis in Dictyostelium by controlling the level of active Rac, Amato et al. have discovered a mechanism that contributes to front-rear polarization in migrating cells. During clathrin-mediated endocytosis, Dictyostelium WASP interacts with active Rac via its CRIB motif. This interaction leads to incorporation of active Rac within endocytic vesicles, which facilitates its removal from inappropriate locations.

Copper oxide nanoparticles (CuO NPs) promoting anticancer activity may be due to the regulation of various classes of histone deacetylases (HDACs). Green-synthesized CuO NPs significantly arrested total HDAC level and also suppressed class I, II and IV HDACs mRNA expression in A549 cells. A549 cells treated with CuO NPs downregulated oncogenes and upregulated tumor suppressor protein expression. CuO NPs positively regulated both mitochondrial and death receptor-mediated apoptosis caspase cascade pathway in A549 cells. Green-synthesized CuO NPs inhibited HDAC and therefore shown apoptosis mediated anticancer activity in A549 lung cancer cell line.

Tumor progression alters the composition and physical properties of the extracellular matrix. Particularly, increased matrix stiffness has profound effects on tumor growth and metastasis. While endothelial cells are key players in cancer progression, the influence of tumor stiffness on the endothelium and the impact on metastasis is unknown. Through quantitative mass spectrometry, we find that the matricellular protein CCN1/CYR61 is highly regulated by stiffness in endothelial cells. We show that stiffness-induced CCN1 activates β-catenin nuclear translocation and signaling and that this contributes to upregulate N-cadherin levels on the surface of the endothelium, in vitro This facilitates N-cadherin-dependent cancer cell-endothelium interaction. Using intravital imaging, we show that knockout of Ccn1 in endothelial cells inhibits melanoma cancer cell binding to the blood vessels, a critical step in cancer cell transit through the vasculature to metastasize. Targeting stiffness-induced changes in the vasculature, such as CCN1, is therefore a potential yet unappreciated mechanism to impair metastasis. ispartof: EMBO Journal vol:36 issue:16 pages:2373-2389 ispartof: location:England status: published

Antimicrobial resistance is one of the major threats to human and animal health. An effective strategy to reduce and/or delay antimicrobial resistance is to use combination therapies. Research in our laboratory has been focused on combination therapies of antimicrobials and phytochemicals and development of antimicrobial-phytochemical conjugates. In this study, we report the synthesis and antimicrobial activity of a novel sulfamethoxazole-gallic acid conjugate compound (Hybrid 1). Hybrid 1 not only showed much stronger activity than sulfamethoxazole towards Streptococcus uberis 19436, Enterococcus faecium 700221 and Enterococcus faecalis 29212, which were purchased from American Type Culture Collection (ATCC), but also exhibited a promising antimicrobial effect against two Enterococcus faecalis clinical isolates, one of which was multidrug resistant. Further studies are warranted to establish the in vivo antimicrobial activity for Hybrid 1 and develop more potent sulfamethoxazole-gallic acid based antimicrobial conjugates using Hybrid 1 as a lead compound.

SummaryEfficient motility requires polarised cells, with anterior pseudopods and a retracting rear. This polarisation requires that the pseudopod catalyst Rac is restricted to the front. Here we show that the Arp2/3 complex regulator WASP is important for maintaining front–rear polarity, using a mechanism that limits where active Rac localises. Dictyostelium cells lacking WASP inappropriately activate Rac and SCAR/WAVE at their rears, leading to reduced cell speed. WASP facilitates the internalisation of clathrin-coated pits, and its Rac-binding CRIB motif is considered essential for its localisation and activity. However, WASP mutants with deleted CRIB domains, or harbouring a new mutation that prevents Rac binding, localise normally, recruit Arp2/3 complex, and drive actin polymerisation. Similarly, Rac inhibitors do not block WASP localisation or activation. Despite this, WASP CRIB mutants cannot restore polarisation of active Rac. Thus, WASP’s interaction with Rac regulates Rac activity and cell polarity, but is dispensable for activating actin polymerization.

Shrimp aquaculture is severely affected by WSSV. Despite an increasing effort to understand host/virus interaction by characterizing changes in gene expression (GE) following WSSV infection, the majority of published studies have focussed on a single time-point, providing limited insight on the development of host-pathogen interaction over the infection cycle. Using RNA-seq, we contrasted GE in gills of Litopenaeus vannamei at 1.5, 18 and 56 hours-post-infection (hpi), between WSSV-challenged and control shrimps. Time course analysis revealed 5097 differentially expressed genes: 63 DEGs were viral genes and their expression in WSSV group either peaked at 18 hpi (and decreased at 56 hpi) or increased linearly up to 56 hpi, suggesting a different role played by these genes during the course of infection. The remaining DEGs showed that WSSV altered the expression of metabolic, immune, apoptotic and cytoskeletal genes and was able to inhibit NF-κB and JAK/STAT pathways. Interestingly, GE changes were not consistent through the course of infection but were dynamic with time, suggesting the complexity of host-pathogen interaction. These data offer novel insights into the cellular functions that are affected during the course of infection and ultimately provide a valuable resource towards our understanding of the host-pathogen dynamics and its variation with time.

In recent years, the importance of viral and host microRNAs (miRNAs) in mediating viral replication and control of host cellular machinery, has been realised and increasing efforts have been taken in order to understand the interactions of miRNAs from host and pathogen during infection. However, all existing studies has thus far been conducted in controlled experimental conditions and the veracity of these data for field conditions are yet to be established. In this framework, small RNA sequencing was performed to identify the miRNAs involved in shrimp (Penaeus vannamei) immune responses under two different WSSV infection conditions of natural infection and experimentally challenged conditions. The expression profiles of miRNAs of shrimp infected with WSSV under two contrasting conditions were compared and as a result, 23365 known miRNAs and 481 novel miRNAs were identified. Amongst the most abundantly expressed miRNAs, the hypoxia related miR-210 and immune pathway related miR-29b were expressed only in infected shrimps of both conditions. miR-8-5p, having a functional role in modulation of chitin biosynthesis was exclusively represented in higher numbers in the WSSV -infected shrimps under natural conditions whilst four of the miRNAs (mja-miR-6493-5p, mja-miR-6492, mmu-miR-3968, tcf-miR-9b-5p) identified from shrimps collected from pond culture targeted chitinase, an important enzyme involved in growth and moulting in shrimps, indicating an interaction between WSSV infection and moult cycle under culture conditions. Some of the miRNAs (tca-miR-87b-3p, cte-miR-277a) and miRNAs belonging to class miR-9, miR-981 that were identified only in WSSV infected shrimps under experimental conditions, are known to respond against WSSV infection in shrimps. Moreover, the miRNA target prediction revealed several immune-related gene targets such as cathepsin, c-type lectin, haemocyanin and ubiquitin protein ligase were commonly identified under both the conditions. However, the miRNAs identified from challenge experiment had wide number of gene targets as compared to the miRNAs of natural infection. The shrimp miRNA mja-miR-6489-3p, was also found to target early virus gene wsv001 of WSSV. Our study, therefore, provides the comparative analysis of miRNA expression from shrimp during WSSV infection in two different conditions.

BRAFV600E is the most common driver mutation in human cutaneous melanoma and is frequently accompanied by loss of the tumor-suppressing phosphatase PTEN. Recent evidence suggests a co-operative role for RAC1 activity in BRAFV600E-driven melanoma progression and drug resistance. However, the underlying molecular mechanisms and the role of RAC1 downstream targets are not well-explored. In this study, we examine the role of the NCKAP1 subunit of the pentameric cytoskeletal SCAR/WAVE complex, a major downstream target of RAC1, in a mouse model of melanoma driven by BRAFV600E;PTEN loss. The SCAR/WAVE complex is the major driver of lamellipodia formation and cell migration downstream of RAC1 and depends on NCKAP1 for its integrity. Targeted deletion of Nckap1 in the melanocyte lineage delayed tumor onset and progression of a mutant Braf;Pten loss‒driven melanoma mouse model. Nckap1-depleted tumors displayed fibrotic stroma with increased collagen deposition concomitant with enhanced immune infiltration. Nckap1 loss slowed proliferation and tumor growth, highlighting a role in cell-cycle progression. Altogether, we propose that NCKAP1-orchestrated actin polymerization is essential for tumor progression and maintenance of tumor tissue integrity in a mutant Braf/Pten loss‒driven mouse model for melanoma.

Background: Bovine mastitis is one of the major infectious diseases in dairy cattle, resulting in large economic loss due to decreased milk production and increased production cost to the dairy industry. Antibiotics are commonly used to prevent/treat bovine mastitis infections. However, increased antibiotic resistance and consumers&rsquo, concern regarding antibiotic overuse make it prudent and urgent to develop novel therapeutic protocols for this disease. Materials and methods: Potential druggable targets were found in 20 mastitis-causing pathogens and conserved and unique targets were identified. Bacterial strains Staphylococcus aureus (ATCC 29213, and two clinical isolates CI 1 and CI 2) and Staphylococcus epidermidis (ATCC 12228, and two clinical isolates CI 1 and CI 2) were used in the present study for validation of an effective drug combination. Results: In the current study, we identified the common and the unique druggable targets for twenty mastitis-causing pathogens using an integrative approach. Furthermore, we showed that phosphorylcholine, a drug for a unique target gamma-hemolysin component B in Staphylococcus aureus, and ceftiofur, the mostly used veterinary antibiotic that is FDA approved for treating mastitis infections, exhibit a synergistic effect against S. aureus and a strong additive effect against Staphylococcus epidermidis in vitro. Conclusion: Based on the data generated in this study, we propose that combination therapy with drugs that work synergistically against conserved and unique targets can help increase efficacy and lower the usage of antibiotics for treating bacterial infections. However, these data need further validations in animal models of infection.

λ genes O and P are required for replication initiation from the bacteriophage λ origin site, oriλ, located within gene O. Questions have persisted for years about whether O-defects can indeed be complemented in trans. We show the effect of original null mutations in O and the influence of four origin mutations (three are in-frame deletions and one is a point mutation) on complementation. This is the first demonstration that O proteins with internal deletions can complement for O activity, and that expression of the N-terminal portion of gene P can completely prevent O complementation. We show that O-P co-expression can limit the lethal effect of P on cell growth. We explore the influence of the contiguous small RNA OOP on O complementation and P-lethality.

White spot syndrome virus is a major pathogen of shrimp, causing economic loss to the aquaculture industry. For the first time, a complete de novo genome of an Indian isolate of this virus has been deciphered using Illumina and Nanopore sequencing technologies. The genome has 280,591 bp with 442 predicted coding genes.

The temperate bacteriophage lambda (λ) CII protein is a positive regulator of transcription from promoter pE, a component of the lysogenic response. The expression of cII was examined in vectors devoid of phage transcription-modulating elements. Their removal enabled evaluating if the expression of the small RNA OOP, on its own, could suppress CII activities, including complementing for a lysogenic response, cell toxicity and causing rapid cellular loss of ColE1 plasmids. The results confirm that OOP RNA expression from the genetic element pO-oop-to can prevent the ability of plasmid-encoded CII to complement for a lysogenic response, suggesting that it serves as a powerful regulatory pivot in λ development. Plasmids with a pO promoter sequence of 45 nucleotides (pO45), containing the −10 and −35 regions for oop, were non-functional; whereas, plasmids with pO94 prevented CII complementation, CII-dependent plasmid loss and suppressed CII toxicity, suggesting the pO promoter has an extended DNA sequence. All three CII activities were eliminated by the deletion of the COOH-terminal 20 amino acids of CII. Host mutations in the hflA locus, in pcnB and in rpoB influenced CII activities. These studies suggest that the COOH-terminal end of CII likely interacts with the β-subunit of RNA polymerase.

Inflammatory and invasive breast cancers are aggressive and require better understanding for the development of new treatments and more accurate prognosis. Here, we detected high expression of PPARα in human primary inflammatory (SUM149PT) and highly invasive (SUM1315MO2) breast cancer cells, and tissue sections of human breast cancer. PPARα ligands are clinically used to treat dyslipidemia. Among lipid lowering drugs clofibrate, fenofibrate and WY14643, clofibrate showed high chemo-sensitivity towards breast cancer cells. Clofibrate treatment significantly induced PPARα DNA binding activity, and remarkably reduced cyclooxygenase-2/PGE2 and 5-lipoxygenase/LTB4 inflammatory pathways. Clofibrate treatment reduced the proliferation of breast cancer cells probably by inhibiting NF-κB and ERK1/2 activation, reducing cyclinD1, cyclinA, cyclinE, and inducing pro-apoptotic P21 levels. Surprisingly, the expression of lipogenic pathway genes including SREBP-1c (sterol regulatory element-binding protein-1c), HMG-CoA synthase, SPTLC1 (serine palmitoyltransferase long-chain), and Acyl-CoA oxidase (ACO) decreased with a concurrent increase in fatty acid oxidation genes such as CPT-1a (carnitine palmitoyltransferase 1a) and SREBP-2 (Sterol regulatory element-binding protein-2). Clofibrate treatment induced secretion of free fatty acids and effectively decreased the level of phosphorylated active form of fatty acid synthase (FASN), an enzyme catalyzing de novo synthesis of fatty acids. High level of coactivators steroid receptor coactivator-1 (SRC-1) and histone acetylase CBP-300 (CREB binding protein-300) were observed in the nuclear complexes of clofibrate treated breast cancer cells. These findings implicate that stimulating PPARα by safe, well-tolerated, and clinically approved clofibrate may provide a safer and more effective strategy to target the signaling, lipogenic, and inflammatory pathways in aggressive forms of breast cancer.

Summary The individual molecular pathways downstream of Cdc42, Rac, and Rho GTPases are well documented, but we know surprisingly little about how these pathways are coordinated when cells move in a complex environment in vivo. In the developing embryo, melanoblasts originating from the neural crest must traverse the dermis to reach the epidermis of the skin and hair follicles. We previously established that Rac1 signals via Scar/WAVE and Arp2/3 to effect pseudopod extension and migration of melanoblasts in skin. Here we show that RhoA is redundant in the melanocyte lineage but that Cdc42 coordinates multiple motility systems independent of Rac1. Similar to Rac1 knockouts, Cdc42 null mice displayed a severe loss of pigmentation, and melanoblasts showed cell-cycle progression, migration, and cytokinesis defects. However, unlike Rac1 knockouts, Cdc42 null melanoblasts were elongated and displayed large, bulky pseudopods with dynamic actin bursts. Despite assuming an elongated shape usually associated with fast mesenchymal motility, Cdc42 knockout melanoblasts migrated slowly and inefficiently in the epidermis, with nearly static pseudopods. Although much of the basic actin machinery was intact, Cdc42 null cells lacked the ability to polarize their Golgi and coordinate motility systems for efficient movement. Loss of Cdc42 de-coupled three main systems: actin assembly via the formin FMNL2 and Arp2/3, active myosin-II localization, and integrin-based adhesion dynamics., Graphical Abstract, Highlights • Loss of Cdc42 in the melanocyte lineage causes pigmentation defects in mice • Cdc42 null melanoblasts are elongated, with severe motility and proliferation defects • Cdc42 and Rac1 have distinct roles for in vivo migration • Cdc42 coordinates actin dynamics, active myosin-II, and integrin-based adhesion, Woodham and Paul et al. describe an important role for the small GTPase Cdc42 in cell proliferation and migration in mouse embryo melanoblasts. Loss of Cdc42 leads to severe pigmentation defects, and reveals Cdc42 as a coordinator of multiple cell-migration systems in the melanocyte lineage, including actin, contractility, and adhesion.

Modulation of gamma-secretase cleavage of Amyloid Precursor Protein (APP) to control the level of Amyloidbeta (A-beta) peptide is one of the strategies to develop therapy for Alzheimer’s disease. Presenilin is a subunit and the catalytic core of gamma-secretase. It has Asp 257 and Asp 385 residues, which are essential for catalytic activity and thus serve as the region of interest for screening of potential gamma-secretase inhibitors. In the present study, in silico screening of drug molecules has been performed in an attempt to identify effective inhibitors of presenilin. Ligand-based pharmacophore models generated with reported inhibitor molecules have been used as query for screening from DrugBank database. Inhibitory activity (IC 50 ) of the screening hits is predicted using a QSAR model developed. The selected molecules have been subjected to docking study against Presenilin1 C-terminal fragment that houses Asp 385 in place of presenilin, as its structure is unavailable. Finally, 46 potential inhibitor molecules were selected based on scores of scoring function and interaction with Asp 385. The selected compounds have spatial arrangement of features essential for binding to presenilin, desired inhibitory activity against processing of APP to A-beta by gamma-secretase and selective interaction with specific amino acids in ligand-protein docked complexes.

Anthropogenic nitrogen contamination has increased in ecosystems around the world (frequently termed the “nitrogen cascade”). Coke production for steel manufacturing is often overlooked as a source of nitrogen to natural ecosystems. We examined sediment cores from a Horseshoe Lake, a floodplain lake located just East of St. Louis Missouri (USA) to test whether a coking plant effluent could be traced using stable isotopes of nitrogen and diatom microfossils. The distribution of δ15N values in surface sediment samples from the lake shows the highest values near the coking plant effluent. Stable isotopes of nitrogen from 4 sediment cores using a mixing model showed three sources of nitrogen since 1688 CE. The first source (active between 1688 and 1920 CE) had a calculated δ15N value ranging between − 0.4 and 1.1‰ depending on the core. After 1920 a second source with a δ15N ranging between 10.6 and 15.4‰ became active. The change in these sources coincides with the construction of a coking plant on the lake shore. A third source with a value approximately 7.0‰ was present at all times and represents background. The diatom microfossil assemblages present from 1688 CE to the late 1800s are dominated by the planktonic species Aulacoseira granulata and periphytic and benthic genera Gomphonema, Cocconeis, and Lyrella. After the late 1800s the diatom assemblages are dominated by Staurosira species indicating a shift of species from high flow riverine environments to epipelic species from a lake environment. Diatom microfossils seem to track the reduction in flooding due to leveeing of the floodplain and the isolation of the lake from the river. Our results show how stable isotopes of nitrogen can be used to track nitrogen inputs from industrial sources. Diatom changes corresponded with changes in connectivity between the Mississippi River and its floodplain.

INTRODUCTION: Plasmodium Vivax malaria is endemic in the northern and eastern part of India. Although Plasmodium falciparum has been commonly implicated as the cause of complicated malaria but now the trend is changing and vivax is increasingly presenting with severe complications. AIMS AND OBJECTIVES: To study the occurrence and severity of thrombocytopenia in patients of vivax malaria. MATERIAL AND METHODS: A total of 100 patients were included in the study and identified positive for malaria parasites on peripheral smear examination with conventional microscopy. Platelet count was done on a fully automated, quantitative, hematology Counter at the time of presentation of patient. INCLUSION CRITERIA: freshly diagnosed cases of vivax malaria. EXCLUSION CRITERIA: Patients with associated falciparum, dengue, leptospirosis or any other known cause of thrombocytopenia. RESULTS: Platelet count was found to be normal in 32 patients. The mean platelet count in vivax malaria was 1,15,390/μl (SD 64,580) with a range of 8000-5,73,000/μl, Mild thrombocytopenia (50000 to 150000) was found in 36 patients. Moderate thrombocytopenia(20000 to 50000) was found in 20 patients. Severe thrombocytopenia (

INTRODUCTION: Tuberculosis remains one of the world’s deadliest diseases & tubercular meningitis is one of the most deadly complications due to missed diagnosis and delayed treatment result in significant morbidity and mortality. The signs and symptoms, results of routine analysis of CSF and radiographic findings for patients with CNS tuberculosis are often inadequate in making a definitive diagnosis. AIMS AND OBJECTIVES: 1. To study the levels of Adenosine deaminase in cases of meningitis, 2. To evaluate the sensitivity and specificity of CSF-ADA as a diagnostic test in tuberculosis meningitis. MATERIAL AND METHODS: In the cross sectional study, Adenosine deaminase activity (ADA) was studied in cerebrospinal fluid of 60 cases of tuberculous meningitis, 10 cases of pyogenic meningitis. 10 cases of aseptic meningitis and15 control. RESULTS: The mean CSF-ADA activity was 14.1±1.96 (tuberculous meningitis); 4.92±1.27 (pyogenic meningitis); 3.66±1.03 (aseptic meningitis) and 1.69±0.44 U/l (control) respectively. The sensitivity and specificity of this test for diagnosis of tuberculous meningitis was 100% and 97.44% respectively with ADA value of more than 10 U/L. CONCLUSION: The adenosine deaminase activity in tuberculous meningitis cases was significantly higher. So it can be of great value in the early and more definitive diagnosis of tuberculous meningitis, to help early diagnosis and treatment to prevent from deadly complications of tubercular meningitis.

Rooting and biomass production of Justicia gendarussa has been achieved through a hydroponic system of cultivation. The obtained biomass of leaves, stem and root were examined for antibacterial activity against various human pathogenic organisms such as Staphylococcus aureus, Escherichia coli, Shigella sp., Pseudomonas sp. and Klebsiella pneumoniae. Methanolic extract of J. gendarussa root responded against E. coli. The growth of Shigella sp., Pseudomonas sp. and K. pneumonia were inhibited by leaf extract. The maximum inhibition zone against S. aureus was observed in stem extract treatment.

The bacteriophage lambda replication initiation protein P exhibits a toxic effect on its Escherichia coli (E. coli) host, likely due to the formation of a dead-end P-DnaB complex, sequestering the replicative DnaB helicase from further activity. Intracellular expression of P triggers SOS-independent cellular filamentation and rapidly cures resident ColE1 plasmids. The toxicity of P is suppressed by alleles of P or dnaB. We asked whether P buildup within a cell can influence E. coli replication fidelity. The influence of P expression from a defective prophage, or when cloned and expressed from a plasmid was examined by screening for auxotrophic mutants, or by selection for rifampicin resistant (Rif(R)) cells acquiring mutations within the rpoB gene encoding the β-subunit of RNA polymerase (RNAP), nine of which proved unique. Using fluctuation assays, we show that the intracellular expression of P evokes a mutator effect. Most of the Rif(R) mutants remained P(S) and localized to the Rif binding pocket in RNAP, but a subset acquired a P(R) phenotype, lost sensitivity to ColE1 plasmid curing, and localized outside of the pocket. One P(R) mutation was identical to rpo*Q148P, which alleviates the UV-sensitivity of ruv strains defective in the migration and resolution of Holliday junctions and destabilizes stalled RNAP elongation complexes. The results suggest that P-DnaB sequestration is mutagenic and supports an earlier observation that P can interact with RNAP.

Objective To study the antidiabetic effects of suspension cell extract of Gymnema sylvestre (G. sylvestre) along with field grown and wild plants. Methods The effect of ethanolic extracts of the in vitro grown suspension cells of G. sylvestre along with field grown and wild plant leaves of G. sylvestre was tested on alloxan induced diabetic rats. Results While oral administration of the extracts reduced the glucose content in blood and urine, sugar and lipids in serum significantly (P≤0.05), it also increased the body weight, total haemoglobin and plasma protein content. Conclusions It can be concluded that G. sylvestre suspension cell extract show excellent antidiabetic potential against alloxan induced diabetic albino male rats therefore be considered as potent antidiabetic drug.

Coronin7 (CRN7) stabilizes F-actin and is a regulator of processes associated with the actin cytoskeleton. Its loss leads to defects in phagocytosis, motility and development. It harbors a CRIB (Cdc42- and Rac-interactive binding) domain in each of its WD repeat domains which bind to Rac GTPases preferably in their GDP-loaded forms. Expression of wild type CRN7 in CRN7 deficient cells rescued these defects, whereas proteins with mutations in the CRIB motifs which were associated with altered Rac binding were effective to varying degrees. The presence of one functional CRIB was sufficient to reestablish phagocytosis, cell motility and development. Furthermore, by molecular modeling and mutational analysis we identified the contact regions between CRN7 and the GTPases. We also identified WASP, SCAR and PAKa as downstream effectors in phagocytosis, development and cell surface adhesion, respectively, since ectopic expression rescued these functions.

Background The receptor for activated C-kinase 1 (RACK1) is a conserved protein belonging to the WD40 repeat family of proteins. It folds into a beta propeller with seven blades which allow interactions with many proteins. Thus it can serve as a scaffolding protein and have roles in several cellular processes. Results We identified the product of the Dictyostelium discoideum gpbB gene as the Dictyostelium RACK1 homolog. The protein is mainly cytosolic but can also associate with cellular membranes. DdRACK1 binds to phosphoinositides (PIPs) in protein-lipid overlay and liposome-binding assays. The basis of this activity resides in a basic region located in the extended loop between blades 6 and 7 as revealed by mutational analysis. Similar to RACK1 proteins from other organisms DdRACK1 interacts with G protein subunits alpha, beta and gamma as shown by yeast two-hybrid, pulldown, and immunoprecipitation assays. Unlike the Saccharomyces cerevisiae and Cryptococcus neoformans RACK1 proteins it does not appear to take over Gβ function in D. discoideum as developmental and other defects were not rescued in Gβ null mutants overexpressing GFP-DdRACK1. Overexpression of GFP-tagged DdRACK1 and a mutant version (DdRACK1mut) which carried a charge-reversal mutation in the basic region in wild type cells led to changes during growth and development. Conclusion DdRACK1 interacts with heterotrimeric G proteins and can through these interactions impact on processes specifically regulated by these proteins.

This study investigates the production of extracellular protease synthesis were carried out by using Aspergillusoryzae was evaluated under different fermentation parameters by employing submerged fermentation method. The protease producers detected by the clear zone (casein hydrolysis) around the colony by simple plate assay method. Aspergillusoryzae KS 5 is the potential strain among the fungal isolates. The Proteasesynthesis were increased their yield after the optimization of fermentation parameters. The optimum pH 6.0, temperature 35 0 C and inoculum size 1.0 ml and it showed 1.7 IU. Key words: Protease, Plate assay, Optimization and Aspergillusoryzae

The Cdc42- and Rac-interactive binding motif (CRIB) of coronin binds to Rho GTPases with a preference for GDP-loaded Rac. Mutation of the Cdc42- and Rac-interactive binding motif abrogates Rac binding. This results in increased 1evels of activated Rac in coronin-deficient Dictyostelium cells (corA(-)), which impacts myosin II assembly. corA(-) cells show increased accumulation of myosin II in the cortex of growth-phase cells. Myosin II assembly is regulated by myosin heavy chain kinase-mediated phosphorylation of its tail. Kinase activity depends on the activation state of the p21-activated kinase a. The myosin II defect of corA(-) mutant is alleviated by dominant-negative p21-activated kinase a. It is rescued by wild-type coronin, whereas coronin carrying a mutated Cdc42- and Rac-interactive binding motif failed to rescue the myosin defect in corA(-) mutant cells. Ectopically expressed myosin heavy chain kinases affinity purified from corA(-) cells show reduced kinase activity. We propose that coronin through its affinity for GDP-Rac regulates the availability of GTP-Rac for activation of downstream effectors.

Background Dictyostelium harbors several paralogous Sec7 genes that encode members of three subfamilies of the Sec7 superfamily of guanine nucleotide exchange factors. One of them is the cytohesin family represented by three members in D. discoideum, SecG, Sec7 and a further protein distinguished by several transmembrane domains. Cytohesins are characterized by a Sec7-PH tandem domain and have roles in cell adhesion and migration. Results We study here Sec7. In vitro its PH domain bound preferentially to phosphatidylinositol 3,4-bisphosphate (PI(3,4)P2), phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3). When following the distribution of GFP-Sec7 in vivo we observed the protein in the cytosol and at the plasma membrane. Strikingly, when cells formed pseudopods, macropinosomes or phagosomes, GFP-Sec7 was conspicuously absent from areas of the plasma membrane which were involved in these processes. Mutant cells lacking Sec7 exhibited an impaired phagocytosis and showed significantly reduced speed and less persistence during migration. Cellular properties associated with mammalian cytohesins like cell-cell and cell-substratum adhesion were not altered. Proteins with roles in membrane trafficking and signal transduction have been identified as putative interaction partners consistent with the data obtained from mutant analysis. Conclusions Sec7 is a cytosolic component and is associated with the plasma membrane in a pattern distinctly different from the accumulation of PI(3,4,5)P3. Mutant analysis reveals that loss of the protein affects cellular processes that involve membrane flow and the actin cytoskeleton.

Kaposi's sarcoma herpesvirus (KSHV) latent oncoprotein viral FLICE (FADD-like interferon converting enzyme)-like inhibitory protein (v-FLIP) or K13, a potent activator of NF-κB, has well-established roles in KSHV latency and oncogenesis. KSHV-induced COX-2 represents a novel strategy employed by KSHV to promote latency and inflammation/angiogenesis/invasion. Here, we demonstrate that v-FLIP/K13 promotes tumorigenic effects via the induction of host protein COX-2 and its inflammatory metabolite PGE2 in an NF-κB-dependent manner. In addition to our previous studies demonstrating COX-2/PGE2's role in transcriptional regulation of KSHV latency promoter and latent gene expression, the current study adds to the complexity that though LANA-1 (latency associated nuclear antigen) is utilizing COX-2/PGE2 as critical factors for its transcriptional regulation, it is the v-FLIP/K13 gene in the KSHV latency cluster that maintains continuous COX-2/PGE2 levels in the infected cells. We demonstrate that COX-2 inhibition, via its chemical inhibitors (NS-398 or celecoxib), reduced v-FLIP/K13-mediated NF-κB induction, and extracellular matrix (ECM) interaction-mediated signaling, mitochondrial antioxidant enzyme manganese superoxide dismutase (MnSOD) levels, and subsequently downregulated detachment-induced apoptosis (anoikis) resistance. vFLIP expression mediated the secretion of cytokines, and spindle cell differentiation activated the phosphorylation of p38, RSK, FAK, Src, Akt and Rac1-GTPase. The COX-2 inhibition in v-FLIP/K13-HMVECs reduced inflammation and invasion/metastasis-related genes, along with reduced anchorage-independent colony formation via modulating 'extrinsic' as well as 'intrinsic' cell death pathways. COX-2 blockade in v-FLIP/K13-HMVEC cells drastically augmented cell death induced by removal of essential growth/survival factors secreted in the microenvironment. Transformed cells obtained from anchorage-independent colonies of COX-2 inhibitor-treated v-FLIP/K13-HMVEC cells expressed lower levels of endothelial-mesenchymal transition genes such as slug, snail and twist, and higher expression of the tumor-suppressor gene, E-cadherin. Taken together, our study provides strong evidences that FDA-approved COX-2 inhibitors have great potential in blocking tumorigenic events linked to KSHV's oncogenic protein v-FLIP/K13.

BackgroundIn eukaryotes the genetic material is enclosed by a continuous membrane system, the nuclear envelope (NE). Along the NE specific proteins assemble to form meshworks and mutations in these proteins have been described in a group of human diseases called laminopathies. Laminopathies include lipodystrophies, muscle and cardiac diseases as well as metabolic or progeroid syndromes. Most laminopathies are caused by mutations in the LMNAgene encoding lamins A/C. Together with Nesprins (Nuclear Envelope Spectrin Repeat Proteins) they are core components of the LINC complex (Linker of Nucleoskeleton and Cytoskeleton). The LINC complex connects the nucleoskeleton and the cytoskeleton and plays a role in the transfer of mechanically induced signals along the NE into the nucleus, and its components have been attributed functions in maintaining nuclear and cellular organization as well as signal transduction.ResultsHere we narrowed down the interaction sites between lamin A and Nesprin-2 to aa 403-425 in lamin A and aa 6146-6347 in Nesprin-2. Laminopathic mutations in and around the involved region of lamin A (R401C, G411D, G413C, V415I, R419C, L421P, R427G, Q432X) modulate the interaction with Nesprin-2 and this may contribute to the disease phenotype. The most notable mutation is the lamin A mutation Q432X that alters LINC complex protein assemblies and causes chromosomal and transcription factor rearrangements.ConclusionMutations in Nesprin-2 and lamin A are characterised by complex genotype phenotype relations. Our data show that each mutation in LMNAanalysed here has a distinct impact on the interaction among both proteins that substantially explains how distinct mutations in widely expressed genes lead to the formation of phenotypically different diseases.

The biosynthesis of silver nanoparticles and its antimicrobial property was studied using bacteria isolated from Spirulina products. Isolated bacteria were identified as Bacillus sp. MSK 1 (JX495945), Staphylococcus sp. MSK 2 (JX495946), Bacillus sp. MSK 3 (JX495947) and Brevundimonas sp. MSK 4 (JX495948). Silver nanoparticles (AgNPs) were synthesized using bacterial culture filtrate with AgNO3. The initial syntheses of Ag nanoparticles were characterized by UV-vis spectrophotometer (by measuring the color change to intense brown). Fourier Transform Infrared Spectroscopy (FTIR) study showed evidence that proteins are possible reducing agents and Energy-dispersive X-ray (EDX) study showing the metal silver as major signal. The structure of AgNPs was determined by Scanning electron microscopy (SEM) and X-ray diffraction (XRD). Synthesized Ag nanoparticles with an average size of 40-65 nm have antimicrobial property against human pathogens like Proteus vulgaris, Salmonella typhi, Vibrio cholera, Streptococcus sp., Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. Among the isolates Brevundimonas sp. MSK 4 alone showed good activity in both synthesis of AgNPs and antimicrobial activity. This work demonstrates the possible use of biological synthesized silver nanoparticles to combat the drug resistant problem.