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While the involvement of protease-activated receptors (PARs) in the physiological regulation of human placenta development, as in tumor biology, is recognized, the molecular pathway is unknown. We evaluated the impact of PAR1 and PAR2 function in cytotrophoblast (CTB) proliferation and invasion in a system of extravillous trophoblast (EVT) organ culture and in human cell-lines. Activation of PAR1 - and PAR2 -induced EVT invasion and proliferation, while the shRNA silencing of low-density lipoprotein receptor-related protein 5/6 (LRP5/6) inhibited these processes. PAR1 and PAR2 effectively induce β-catenin stabilization in a manner similar to that shown for the canonical β-catenin stabilization pathway yet independent of Wnts. Immunoprecipitation analyses and protein-protein docking demonstrated the co-association between either PAR1 or PAR2 with LRP5/6 forming an axis of PAR-LRP5/6-Axin. Noticeably, in PAR1 -PAR2 heterodimers a dominant role is assigned to PAR2 over PAR1 as shown by inhibition of PAR1 -induced β-catenin levels, and Dvl nuclear localization. This inhibition takes place either by shRNA silenced hPar2 or in the presence of a TrPAR2 devoid its cytoplasmic tail. Indeed, TrPAR2 cannot form the PAR1 -PAR2 complex, obstructing thereby the flow of signals downstream. Elucidation of the mechanism of PAR-induced invasion contributes to therapeutic options highlighting key partners in the process.

The large variation in day and night temperature of Thar Desert area provides an opportunity to reduce temperature fluctuations and thermal discomfort inside temporary shelters and buildings using ...

Estrogen is reported to have multi-dimensional neuroprotective activity against cognitive dysfunctions in animals and human's despite of its classical hormonal effects. It has been well documented that postmenopausal women exhibit a higher risk of cognitive impairment than premenopausal females. In this context, estrogen replacement therapy (ERT) has been established as one of the better alternative therapy to alleviate cognitive deficits in such females. As per Women's Health Initiative, though ERT is safe and effective strategy to mitigate cognitive dysfunction in postmenopausal ladies still it is reported to have a risk of developing breast and endometrial cancer in these women. Thus, there is a demand for developing suitable alternative medicine to conventional estrogen replacement therapy in the management of cognitive deficits. Recent phytopharmacological research suggests that phytoestrogens, naturally occurring non-steroidal compounds, from plants especially in soy and soy-based products exhibits estrogen-like physiological and pharmacological actions. Phytoestrogens primarily act through binding to estrogenic receptors (ERs) α and β, with a higher affinity for ERβ. Isoflavone phytoestrogens such as Daidzein and Genistein are widely studied herbal drugs in the management of cognitive dysfunction in postmenopausal women among other classes including lignans and coumestans. Experimental studies report that phytoestrogens exert neuroprotective activity with anti-tumor property to uterine growth. Further, clinical and in-vitro studies also corroborate the fact that phytoestrogens exhibit minimal anti-cancer activity. Epidemiological survey also documents that Asian population who consume phytoestrogen-rich diet have a lower risk of breast and prostate cancer compared to western individuals. Hence, it can be presumed that phytoestrogens could be used as a safe and alternative medicine to estrogen replacement therapy in the management of cognitive deficits in postmenopausal ladies.

The essential role of G-protein coupled receptors (GPCRs) in tumor growth is recognized, yet a GPCR based drug in cancer is rare. Understanding the molecular path of a tumor driver gene may lead to the design and development of an effective drug. For example, in members of protease-activated receptor (PAR) family (e.g., PAR1 and PAR2), a novel PH-binding motif is allocated as critical for tumor growth. Animal models have indicated the generation of large tumors in the presence of PAR1 or PAR2 oncogenes. These tumors showed effective inhibition when the PH-binding motif was either modified or were inhibited by a specific inhibitor targeted to the PH-binding motif. In the second part of the review we discuss several aspects of some cardinal GPCRs in tumor angiogenesis.

Purpose The purpose of this study is to improve the anticorrosion performance of low nickel stainless steel (AISI 201) in 3.5% NaCl by electroactive polyimide/copper oxide (EPI/CuO) composites coating. Design/methodology/approach Electroactive polyimide/copper oxide (EPI/CuO) composites were prepared by oxidative coupling polymerization followed by thermal imidization method. Findings The functional and structural properties of composites were characterized by X-ray diffraction, Fourier transmission infra-red and ultra violet-visible spectroscopy and the surface topography was characterized by field emission scanning electron microscope analysis and anticorrosion performance in 3.5 Wt.% NaCl was evaluated by electrochemical techniques. The obtained results of electrochemical techniques measurement indicated that the composites coated samples give better corrosion protection against attacking electrolyte. Originality/value The ever-increasing price of nickel (Ni) is driving the industries to use low-Ni austenitic stainless steels (ASSs). However, it exhibits relatively poor corrosion resistance as compared with conventional Cr-Ni ASSs. Nonetheless, its corrosion resistance can be enhanced by polymeric (electroactive polyimide [EPI]) coating. CuO particles exhibit the hydrophobic properties and can be used as inorganic filler to incorporate in EPI to further enhance the corrosion protection. The present research paper is beneficial for industries to use low-cost AISI 201, enhance its corrosion resistance and replace the use of costly conventional Cr-Ni ASSs.

Multidrug resistance (MDR) is a major health issue for the treatment of infectious diseases throughout the world. Staphylococcus aureus (S. aureus) is a Gram-positive bacteria, responsible for various local and systemic infections in humans. The continuous and abrupt use of antibiotics against bacteria such as S. aureus results in the development of resistant strains. Presently, mupirocin (MUP) is the drug of choice against S. aureus and MDR (methicillin-resistant). However, S. aureus has acquired resistance against MUP as well due to isoleucyl-tRNA synthetase (IleS) mutation at sites 588 and 631. Thus, the aim of the present study was to discover novel bioactives against MUP-resistant S. aureus using in silico drug repurposing approaches. In silico drug repurposing techniques were used to obtain suitable bioactive lead molecules such as buclizine, tasosartan, emetine, medrysone, and so on. These lead molecules might be able to resolve this issue. These leads were obtained through molecular docking simulation based virtual screening, which could be promising for the treatment of MUP-resistant S. aureus. The findings of the present work need to be validated further through in vitro and in vivo studies for their clinical application.

In silico molecular docking is an efficient technique for drug design that predicts the optimized orientation of the ligand against a specific drug target. This is a cost-effective and tim...

Naproxen (NAP) is one of the commonly used nonselective Cycloxygenase (COX) inhibitors. It is a choice of drug for anti-inflammatory activity by subsiding the generation of the inflammatory components called prostaglandins. The common problem associated with the NAP is gastrointestinal toxicity. It may cause ulceration or stomach bleeding. In this study, the different derivatives of NAP were designed by using phytophenols with the aim that they exert the antioxidant activity and have the potential to reduce ulcer formation. The lead molecules were designed by molecular docking-based virtual screening against human COX-2 enzyme through AutoDock. Then these derivatives were screened for pharmacokinetic profiling by considering Lipinski's filter. The potent and safe molecule was identified by pharmacokinetics and toxicity evaluation. The potent compound was synthesized in the laboratory, purified, characterized, and its pharmacological activities were evaluated. The resultant compound was found to be equipotent and less toxic than the parent compound.

Here, we report the experimental observation of multiferrocity in vanadium doped ZnO. The wurtzite ZnO is non-centrosymmetric and hence should be ferroelectric. However, ferroelectricity in ZnO has not been observed yet, most probably due to large barrier between two polarized states. It may be possible to reduce the barrier by doping ions whose size is larger than Zn. It has been attempted to engineer ferroelectricity in transition metal ion doped ZnO. Vanadium could be a possible candidate for this purpose as size of V (0.88A) is much larger than that of Zn (0.74 A) and could result substantial strain inside ZnO. X-ray diffraction result revealed that 5% V-doped ZnO crystallizes in hexagonal wurtzite structure with inbuilt strain. As explained, we indeed observe 5% V-doped ZnO show ferromagnetism and ferroelectricity at room temperature.

Background: Nitric oxide (NO) is an effective mediator of ischemic preconditioning (IPC)-induced cardioprotection. Atrial natriuretic peptide (ANP) is downregulated after ovariectomy, which results in reduction in the level of NO. The present study deals with the investigation of the role of ANP in abrogated cardioprotective effect of IPC in the ovariectomized rat heart. Methods: Heart was isolated from ovariectomized rat and mounted on Langendorff’s apparatus, subjected to 30 min of ischemia and 120 min of reperfusion. IPC was given by four cycles of 5 min of ischemia and 5 min of reperfusion with Krebs–Henseleit solution. The myocardial infract size was estimated employing triphenyltetrazolium chloride stain, and coronary effluent was analyzed for creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) release to consider the degree of myocardial injury. The cardiac release of NO was estimated by measuring the level of nitrite in coronary effluent. Results: IPC-mediated cardioprotection was significantly attenuated in ovariectomized rat as compared to normal rat, which was restored by perfusion with ANP. However, this observed cardioprotection was significantly attenuated by perfusion with L-NAME, an endothelial nitric oxide synthase inhibitor, and Glibenclamide, a KATP channel blocker, alone or in combination noted in terms of increase in myocardial infract size, release of CK-MB and LDH, and also decrease in release of NO. Conclusion: Thus, it is suggested that ANP restores the attenuated cardioprotective effect of IPC in the ovariectomized rat heart which may be due to increase in the availability of NO and consequent increase activation of mitochondrial KATP channels.

Objective Ischemic postconditioning (IPOC) is an endogenous cardioprotective phenomenon, which gets attenuated during the hyperlipidemia. It has been documented that erythropoietin (EPO), a glycoprotein, produces IPOC-like cardioprotection through common signaling pathway such as PI-3K pathway. The aim of this study has been designed to investigate the role of EPO in IPOC induced cardioprotection in hyperlipidemic rat heart. Materials and methods Heart was isolated from hyperlipidemic rat and mounted on Langendorff's apparatus, subjected to 30 min ischemia and 120 min reperfusion. IPOC was mediated by four cycles of 5 min reperfusion and 5 min ischemia. The infarct size was estimated using TTC stain and coronary effluent was analyzed for LDH and CK-MB release to assess the degree of myocardial injury. Results Four cycles of IPOC produces cardioprotection noted in terms of decrease in infarct size and decrease in the release of LDH and CK-MB in normal rat heart. However, IPOC-induced cardioprotection was completely attenuated in isolated heart obtained from hyperlipidemic rat. Perfusion of EPO (1 U/ml) significantly restored the attenuated cardioprotection in hyperlipidemic rat heart, which was completely blocked by perfusion of LY294002 (10 μM), a selective inhibitor of PI-3K. Conclusion Thus, it is suggested that EPO restores the attenuated cardioprotective effect of IPOC in hyperlipidemic rat heart by the activation of PI-3K signaling pathway.

The study deals with the synthesis and evaluation of some novel triazoles. Amine linked triazoles were prepared through multistep syntheses. 4-Iodobenzoic acid was taken as initial reactant material, which was converted into methyl ester, then to hydrazide and finally into triazole via dithiocarbazate salt. The triazole was treated with chloroacetyl chloride and further with secondary amines to get the title compounds. Structures of the synthesized compounds were elucidated by Fourier transform infrared spectroscopy (FTIR), 13C and 1H nuclear magnetic resonance (NMR), mass spectrometry and elemental analysis. The synthesized compounds were screened for their antimicrobial activity. The study was carried out against four bacteria and three fungi by disc diffusion method using ciprofloxacin and fluconazole as reference drug for antibacterial and antifungal activities respectively. All the synthesized compounds exhibited zones of inhibition against Candida albicans, Aspergillus niger and Fusarium oxysporum, but the inhibition was more prominent as shown by compound T83. Similarly antibacterial activity was studied against Staphylococcus aureus, Basillus subtilis, Psedomonas aeruginosa and Escherichia coli. Data revealed that compounds T81 and T83 were effective against the bacterial strains. Minimum inhibitory concentration test (MIC) was also carried out to ascertain the bacteriostatic property of the test compounds.

The present work deals with the pharmacological screening of some newly synthesized triazoles. A series of 1,2,4-triazoles have been synthesized using benzoic acid or 4-chloro benzoic acid as the starting materials. The synthesized compounds were characterized by physical and spectral analysis viz., Fourier transform infrared spectroscopy, 1H nuclear magnetic resonance, 13C nuclear magnetic resonance, Gas Chromatography-Mass Spectrometry and elemental analysis Carbon, Hydrogen and Nitrogen analysis in order to confirm the structure. Acute toxicity studies were carried out in accordance with the Organization for Economic Co-operation and Development guideline 425. The compounds were not found to be lethal even at a dose level of 2000 mg/kg. Pharmacological evaluation was done following three intact animal experiments and one experiment on the isolated tissue. Results of the study indicated that the compound 7bi and 7bj protected up to 60% against histamine-induced dyspnea. Antihistaminic nature of the test compounds 7bi, 7bj, 7ai, and 7bk were also confirmed by the loss of catalepsy after the administration of clonidine (1%, s.c.). During experiments on isolated tissue, suppression of dose-response curve of histamine indicates a noteworthy denouement in favor of the said effect.

Hyperhomocysteinemia (Hhcy) is a biochemical alteration with plasma levels of homocysteine higher than 15 µmol/L, associated with atherosclerosis, and with vascular thrombosis by disrupting endothelial cells. Homocysteine is a sulfur-containing amino acid derived from methionine which is an essential amino acid. Excess homocysteine produced in the body is expelled out by liver and kidney from the systemic circulation. Hhcy is caused by the excess deficiencies of the vitamins like pyridoxine (B6), folic acid (B9), or cyanocobalamin (B12). High protein consumers are usually at risk for hyperhomocysteinemia because of low plasma B12 levels. It is approximated that mild Hhcy occurs in 5–7% of the general population and 40% in patients with vascular disease. Patients with heart failure, impaired renal function, and diabetes should be screened since the prevalence of Hhcy in these patients appears to be quite high. In this article, we hypothesise that citicoline is a novel drug for the management of Hhcy. Furthermore, the side effects of citicoline are also minimal and self-limiting. If this strategy is validated, citicoline will be the cost-effective way to be administered for Hhcy. Many evidences are available which suggest that ignoring homocysteine levels in patients with the vascular disease would be unwise. Thus, there is an urgent need for health care providers to develop effective preventions and interventions program (folic acid, Vitamin B6 and Vitamin B12 supplementation as well as lifestyle change) to reduce this disorder.

This study aims at the development of composite surface on austenitic stainless (AISI 304 stainless steel) by laser surface alloying with WC and Co in the weight ratio of 4:1. Laser processing has been carried out by laser melting of sandblasted AISI 304SS using a 5 kW continuous-wave Nd: YAG laser having a beam diameter of 3 mm and simultaneous addition of powder (WC and Co in the ratio of 4:1) at a rate of 10 mg/sec using He as shroud (at a flow rate of 6 l/min) in the melted surface. The process parameters are applied power (1.75–2 kW) and scan speed (12–30 mm/sec) with a powder feed rate of 10 mg/s. The laser-treated surface has been subjected to microstructural investigation and wear resistance (fretting wear behaviour) testing. There is formation of different carbides (WC, W2C, Fe7C3, Cr23C6, Fe6W6C, and Co6W6C) in the gamma matrix of the treated zone. The microhardness of composite surface is significantly higher (980 VHN) than that of AISI 304SS (242 VHN). The fretting wear kinetics and coefficient of friction (COF) of the composite surface are reduced as compared to the substrate. The mechanism of wear has been established.

Tuberculosis is one of the main causes of morbidity and mortality across the globe. Mycobacterium tuberculosis can attack any part of the human body and can cause treacherous effects also. It maltr...

Background It has been noted that nitric oxide (NO) is involved in the ischemic preconditioning (IPC)–mediated cardioprotection. Diabetes is a downregulator of atrial natriuretic peptide (ANP), resulting in low expression of endothelial nitric oxide synthase (eNOS) by which NO level get reduced. The purpose of the present study was to investigate the role of ANP in attenuated cardioprotective effect of IPC in the diabetic rat heart. Methods The heart was isolated from the diabetic rat and mounted on Langendorff's apparatus, subjected to 30-min ischemia and 120-min reperfusion. IPC was mediated by four cycles of 5-min ischemia and 5-min reperfusion. The infarct size was estimated using triphenyltetrazolium chloride stain, and coronary effluent was analyzed for lactate dehydrogenase and creatinine kinase-MB release to assess the degree of myocardial injury. The cardiac release of NO was estimated indirectly by measuring the release of nitrite in coronary effluent. Results IPC-mediated cardioprotection was significantly attenuated in the diabetic rat as compared with the normal rat. Perfusion of ANP (0.1 μM/L) in the diabetic rat heart significantly restored the attenuated cardioprotective effect of IPC and also increased the release of NO. However, this observed cardioprotection was significantly attenuated by perfusion of N-nitro L-arginine methyl ester, an eNOS inhibitor (100 μM/L) noted in terms of increase in myocardial infarct size, release of lactate dehydrogenase and creatinine kinase-MB, and also decreases in release of NO. Conclusions Thus, it is suggested that ANP restores the attenuated cardioprotective effect in the diabetic heart which may be due to increase in the expression of eNOS and subsequent increase in the activity of NO.

This study demonstrates a dose-dependent response of Trichoderma harzianum Th-56 in improving drought tolerance in rice by modulating proline, SOD, lipid peroxidation product and DHN / AQU transcript level, and the growth attributes. In the present study, the effect of colonization of different doses of T. harzianum Th-56 strain in rice genotypes were evaluated under drought stress. The rice genotypes treated with increasing dose of T. harzianum strain Th-56 showed better drought tolerance as compared with untreated control plant. There was significant change in malondialdehyde, proline, higher superoxide dismutase level, plant height, total dry matter, relative chlorophyll content, leaf rolling, leaf tip burn, and the number of scorched/senesced leaves in T. harzianum Th-56 treated rice genotypes under drought stress. This was corroborated with altered expression of aquaporin and dehydrin genes in T. harzianum Th-56 treated rice genotypes. The present findings suggest that a dose of 30 g/L was the most effective in improving drought tolerance in rice, and its potential exploitation will contribute to the advancement of rice genotypes to sustain crop productivity under drought stress. Interaction studies of T. harzianum with three aromatic rice genotypes suggested that PSD-17 was highly benefitted from T. harzianum colonization under drought stress.

In this manuscript, we describe various approaches that we have examined towards the total synthesis of Cytospolide E. We initially attempted the RCM approach employing first and second generation Grubbs and Grubbs–Hoyeda catalysts resulting in the exclusive synthesis of the Z -isomer of Cytospolide E. With the Furstner catalyst, the dimerization involving the less hindered olefin was the exclusive event. Alternative approach documented is a successful cross-metathesis leading to a seco -acid with the requisite E -configuration and undesired macrodiolide formation during the attempted Shiina's lactonization.

Water pollution is a major concern influencing human lives in many ways. It affects not only the nature of potable water, but also the health of those who consume it. Hazardous water pollutants are broadly categorized into soluble toxicants and toxic planktons. The presence of planktons in potable water causes a negative impact on the human body. The inadmissible planktons of potable water such as copepods, protozoans, helminths, anilids, blue-green algae, dinoflagellates, and diatoms can lead to severe dysfunction in the normal physiology of humans. Complications of the gastrointestinal tract, hepatobiliary system and brain result from drinking polluted water. Toxins such as cyanobacterial toxin, saxitoxin, neosaxitoxin and domoic acid are by-products of the inadmissible planktons which cause adverse effects on the human organ systems. They also derange the percentage of dissolved oxygen in the water bodies. The health-related consequences of the planktons range from cell damage to mortality.

Pesticide resistance and environment threat due to injudicious use of chemical pesticides for disease management employs the alteration in management practices. Chitosan, a deacetylated chitin derivative, behaves like a general elicitor, inducing a non-host resistance, and prime the plants for systemic acquired resistance in addition to this Chitosan has high antimicrobial activity against a wide range of pathogenic and spoilage microorganisms, including fungi and bacteria. The use of chitosan in agriculture and in food systems should be based on sufficient knowledge of the complex mechanisms of its elicitor and antimicrobial mode of action. In this article we a number of studies on the investigation of chitosan antimicrobial and resistance inducing properties and application of them in agriculture sector have been summarized.

The physics of Carbon Nanotubes has rapidly evolved into a research field since their discovery by lijima (4) in multiwall form in 1991. In this research we have attempted an approach to obtain homogeneously dispersed CNTs in Al powders for preparing Al/CNTs nanocomposite. Ball milled Al and CNTs mixture has been compacted by universal testing machine under the load of 3 tons. Micro hardness and relative density of compacted composites of different weight percentage ratio of CNTs has been measured. By using Vicker hardness test, Optical Microscope, Scanning Electron Microscope (SEM) techniques we analyzed the hardness, micro structures and grain boundaries of casted Al-CNTs composites and compared the hardness of pure Al and CNTs reinforced Al composites at different weight percentages of CNTs. Also we checked the mixing pattern of Al CNT composites using X-Ray Diffraction (XRD) technique.

Tin oxide, SnO2 nanoparticles was synthesized by using ethylene glycol as solvent, urea as precipitant and tin metal solution. Electroactive polyimide, EPI/SnO2 nanocomposites with different weight percentages of SnO2 nanoparticles were produced by using insitu oxidative coupling polymerization of oligoaniline in presence of SnO2 nanoparticles followed by thermal imidization. The structural, morphological and thermal characterizations of all samples were performed by XRD, FTIR, SEM, TEM and TGA. The relative anticorrosive properties of coated EPI/SnO2 nanocomposites were investigated by means of potentiodynamic polarization method in NaCl (5%) solution. The thermal stability of EPI/SnO2 nanocomposite shows higher thermal stability compared to EPI this may be due to incorporation of SnO2 in the EPI matrix. EPI/SnO2 nanocomposites coated 316L stainless steel and cold rolled steel samples indicates better anticorrosion properties compared to bare steels and EPI coating.

A sweet disposition: Simple protocols for the synthesis of (1→1)- and (1→6)-linked C-disaccharides have been developed (see scheme). A one-pot Au-catalyzed alkynol cycloisomerization followed by BF3⋅Et2O, Et33SiH reduction has been established as the key reaction for C-disaccharide synthesis including all possible pyran–pyran, furan–furan, and pyran–furan combinations.

Biological control or biocontrol is the use of specific microorganisms that interfere with plant pathogens and pests, and sustain organisms useful to human. It is a nature-friendly, ecological approach to overcome the problems caused by standard chemical methods of plant protection which drastically affect the environment as well as consumer. Trichoderma, a member of ascomycota was first described by Pearson (1794) and its beneficial activities as a biocontrol agent (BCA) has been known since 1930 and since then there have been extensive efforts to use them for plant disease control. Trichoderma species play an important role in controlling fungal plant pathogens, especially the soil borne pathogens, by competing for nutrients and space, producing cell wall degrading enzymes (CWDE) or through mycoparasitism (the direct attack of one fungus on another). They are not only fungal parasites but can also produce antibiotics. These processes such as the production of antifungal metabolites and hydrolytic enzymes are mediated by G-proteins and mitogen activated protein (MAP) kinases. In addition, certain strains can release a variety of compounds that induce systemic and localized resistance response in host plant, mediated by alteration in plant gene expression. This Trichoderma mediated induced systemic resistance (ISR) is regulated by various plant hormones like jasmonic acid (JA), ethylene (ET), abscisic acid (ABA), gibberellic acid (GA) and salicylic acid (SA). Among them, JA, ET and SA are the central players in defence signalling. Thus, Trichoderma encompasses different mechanisms to achieve effective disease control against a variety of plant pathogens by means of various signalling components and pathways. Another approach for improving resistance against pathogens is to express genes of Trichoderma in plants, through genetic manipulation. Recently, there has been increasing attempt in development of transgenic plants using genes from Trichoderma spp. which are responsible for biocontrol activity. Besides protecting against biotic stress, root-colonization with Trichoderma also enhances growth and productivity of host plant by improving nutrient and water uptake. Due to the effective control of plant pathogens/diseases and improvement of plant growth, several Trichoderma bioproducts are now available commercially. However, much work still needs to be done to explore its full potential for improving plant growth and managing plant diseases.

A series of fifteen new substituted 1,2,4-triazoles (6a – 6o) have been synthesized in order to obtain new agents with potential anti-inflammatory and anti-nociceptive activity. The title compounds were synthesized using 4-methylbenzoic acid as a starting reactant. The synthesized compounds were characterized by spectroscopic methods (IR, 1H NMR, 13 C NMR, FAB+ − MS) and elemental analysis (nitrogen analysis). The title compounds were evaluated for anti-inflammatory activity by the carrageenan induced paw edema method, and some compounds were evaluated for anti-nociceptive activity by the hot plate method and tail immersion method. Compounds N-[3-mercapto-5-(4-methylphenyl)-4H-1,2,4-triazol-4-yl]-2-piperidin-1-ylacetamide (6f), 2-(4-benzylpiperazin-1-yl)-N-[3-mercapto-5-(4-methylphenyl)-4H-1,2,4-triazol-4-yl]acetamide (6i) and N-[3-mercapto-5-(4-methylphenyl)-4H-1,2,4-triazol-4-yl]-2-morpholin-4-ylacetamide (6 k) showed significant anti-inflammatory activity compared to other synthesized compounds, and N-[3-mercapto-5-(4-methylphenyl)-4H-1,2,4-triazol-4-yl]-2-morpholin-4-ylacetamide (6 k) exhibited superior anti-nociceptive activity.

Lymphatic filarial nematodes maintain a mutualistic relationship with the endosymbiont Wolbachia . Depletion of Wolbachia produces profound defects in nematode development, fertility, and viability and thus has great promise as a novel approach for treating filarial diseases. NAD + -dependent DNA ligase is an essential enzyme of DNA replication, repair, and recombination. Therefore, in the present study, the antifilarial drug target potential of the NAD + -dependent DNA ligase of the Wolbachia symbiont of Brugia malayi ( w Bm-LigA) was investigated using dispiro-cycloalkanone compounds. Dispiro-cycloalkanone specifically inhibited the nick-closing and cohesive-end ligation activities of the enzyme without inhibiting human or T4 DNA ligase. The mode of inhibition was competitive with the NAD + cofactor. Docking studies also revealed the interaction of these compounds with the active site of the target enzyme. The adverse effects of these inhibitors were observed on adult and microfilarial stages of B. malayi in vitro , and the most active compounds were further monitored in vivo in jirds and mastomys rodent models. Compounds 1, 2, and 5 had severe adverse effects in vitro on the motility of both adult worms and microfilariae at low concentrations. Compound 2 was the best inhibitor, with the lowest 50% inhibitory concentration (IC 50 ) (1.02 μM), followed by compound 5 (IC 50 , 2.3 μM) and compound 1 (IC 50 , 2.9 μM). These compounds also exhibited the same adverse effect on adult worms and microfilariae in vivo ( P < 0.05). These compounds also tremendously reduced the wolbachial load, as evident by quantitative real-time PCR ( P < 0.05). w Bm-LigA thus shows great promise as an antifilarial drug target, and dispiro-cycloalkanone compounds show great promise as antifilarial lead candidates.

Objective: The objective of this work is to synthesize and evaluate some novel 1,2,4-triazoles.Methods: Procedure includes synthesis of triazole compounds followed by biological evaluations. The synthesis was carried out in six steps withp-bromobenzoic acid as starting material and converting to ester and then to hydrazide. Hydrazide was then converted to 4-amino triazole. Theamino triazole was then linked to different secondary amines using chloroacetyl chloride as the linking agent. All the synthesized compounds werecharacterized through Fourier transform infrared spectroscopy, gas chromatography-mass spectroscopy, and nuclear magnetic resonance. Further,the compounds were taken out for biological evaluations. To explore their effects, experiments were conducted on various micro- as well as macroorganisms.The toxicityprofilewasalso testedin accordancewith OECD 425 guideline on Wistar albino rats.Results: The compounds were examined for antibacterial as well as antifungal activities. Among the all compound T71, T73, and T75 exhibitedantibacterial activity, and compound T71 showed antifungal activity as well. The evaluation was also carried out for anthelmintic activities. Thecompounds were treated on Pheretima posthuma at various concentrations to explore their vermifuge and vermicidal action. The triazole linked with1-methylpiperazine was found to have comparable activity to that of reference standards.Conclusion: Triazoles are a most potent assemblage of fungal retardants. But depending on their substituents, they also have diverse pharmacologicalvalues. In this study, the compound T71 showed promising antimicrobial as well as anthelmintic action. Hence, it can be considered as a lead compoundfor further researches.

Objectives Wolbachia, an endosymbiont of filarial nematode, is considered a promising target for therapy against lymphatic filariasis. Transcription elongation factor GreA is an essential factor that mediates transcriptional transition from abortive initiation to productive elongation by stimulating the escape of RNA polymerase (RNAP) from native prokaryotic promoters. Upon screening of 6257 essential bacterial genes, 57 were suggested as potential future drug targets, and GreA is among these. The current study emphasized the characterization of Wol GreA with its domains. Methodology/Principal Findings Biophysical characterization of Wol GreA with its N-terminal domain (NTD) and C-terminal domain (CTD) was performed with fluorimetry, size exclusion chromatography, and chemical cross-linking. Filter trap and far western blotting were used to determine the domain responsible for the interaction with α2ββ′σ subunits of RNAP. Protein-protein docking studies were done to explore residual interaction of RNAP with Wol GreA. The factor and its domains were found to be biochemically active. Size exclusion and chemical cross-linking studies revealed that Wol GreA and CTD exist in a dimeric conformation while NTD subsists in monomeric conformation. Asp120, Val121, Ser122, Lys123, and Ser134 are the residues of CTD through which monomers of Wol GreA interact and shape into a dimeric conformation. Filter trap, far western blotting, and protein-protein docking studies revealed that dimeric CTD of Wol GreA through Lys82, Ser98, Asp104, Ser105, Glu106, Tyr109, Glu116, Asp120, Val121, Ser122, Ser127, Ser129, Lys140, Glu143, Val147, Ser151, Glu153, and Phe163 residues exclusively participates in binding with α2ββ′σ subunits of polymerase. Conclusions/Significance To the best of our knowledge, this research is the first documentation of the residual mode of action in wolbachial mutualist. Therefore, findings may be crucial to understanding the transcription mechanism of this α-proteobacteria and in deciphering the role of Wol GreA in filarial development., Author Summary Diethylcarbamazine and ivermectin, the conventional antifilarial drugs, exert poor activity on adult filarial worms. Brugia malayi, a causative agent of human lymphatic filariasis, harbors an obligate alpha-proteobacterium, Wolbachia. Because the endosymbiont is required for the development, viability, and fertility of the parasite, wolbachial proteins seem to be a potential target for antifilarial drugs. Transcription elongation factor is one of the bacterial factors that plays an essential role in efficient transcription by facilitating the movement of RNA polymerase on DNA template. The factor is also necessary for bacterial viability because deletion of greA leads to hypersensitivity to environmental assaults such as ionic detergents, elevated temperature or osmotic shock. Functional characterization of Wol GreA with its domains was undertaken to ascertain the molecular trappings underlying Wolbachia maintenance within the filarial parasite and gain further insight into the mutualism between the bacteria and the nematode host. In the present study, GreA of Wolbachia revealed phylogenetic divergence from other bacteria. Wol GreA, a member of the Gre family, authenticates its essentiality by the presence of evolutionarily conserved N and C terminal domains, where dimeric CTD exhibited chemical cross-linking and direct binding to α2ββ′σ subunits of Wol RNAP.

Introduction: 1,2,4-triazoles and its derivatives have been reported to possess anti-inflammatory, analgesic, antimicrobial, anticancer, antitumor, antitubercular, anticonvulsant, openers of Ca-activated potassium (Maxi-K) channels, antiviral properties, hypoglycemic, anxiolytic and antidepressant activity. Therefore, 1,2,4-triazole seems to be an important pharmacophore. Materials and Methods: The synthesis of 4-(substituted ethanoyl) amino-3-mercapto-5-(4-methoxy) phenyl-1,2,4-triazoles (6a-o) were prepared following six step starting 4-methoxy benzoic acid and using different secondary amines and were characterized with the help of FT-IR, 1 H, 13 C NMR, FAB Mass and nitrogen analysis. These synthesized compounds (6a-o) were then evaluated for anti-inflammatory activity by carrageenan induced paw edema method.Out of these synthesized compounds, some (6f, i and k) were evaluated for antinociceptive activity by Hot plate method and Tail immersion method. Results and Discussion: The synthesis of 4-(substituted amino)-3-mercapto-5-(4-methoxy) phenyl-1,2,4-triazoles (6a-o) was accomplished. The IR spectra exhibited characteristic bands for C-N, C=N, SH and C=O at 1350-1360, 1511-1548, 2520-2594.3 and 1650-1719 cm -1 . The C-O-C asymmetric and symmetric str. was at 1250-1254 and 1027-1079.3 cm -1 respectively. In 1 H-NMR spectra, a singlet of CONH was found in the range of δ 9.92-10.18 ppm and another singlet of thiol group was observed in the range of δ 8.63-9.92 ppm. A singlet of Ar-OCH 3 was also found between δ 3.57-3.91 ppm. In 13 C- NMR spectra, C-3 and C-5 of the 1,2,4 - triazole nucleus were observed in the range of δ 147-166.9 ppm. Carbonyl carbon and methylene carbon of -NHCOCH 2 N< were found between δ 166.5-177.5 and δ 47.1-62 ppm respectively. Acute toxicity study was donr following OECD-423 and cut-off dose was found to be between 1000-1500 mg/kg body weight. At the dose level of 100 mg/kg, 6f, 6i and 6k exhibited appreciable inhibition of oedema especially 6k exhibiting a percentage of oedema inhibition of 40.28%, which was comparable to that of the standard drug indomethacin (62.50% at 10mg/kg dose). Among the compounds tested, compound 6k exhibited good anti-nociceptive activity in both methods used. Pethidine (20mg/kg body weight s.c) is used as the standard drug. Conclusion: SAR of these synthesized compounds shows that substitution with heterocyclic moiety at C-2 of the acetamido group at position 4 of the 1,2,4-triazole produces appreciable activity as compared to substitution with aliphatic moieties since among all the synthesized compounds, the most active ones are 6f, 6i and 6k that have piperdine, 1-benzyl piperazine and morpholine group, respectively at C-2 of the acetamido group at position 4 of the 1,2,4-triazole.

The lymphatic filarial parasite, Brugia malayi contains Wolbachia endobacteria that are essential for development, viability and fertility of the parasite. Therefore, wolbachial proteins have been currently seen as the potential antifilarial drug targets. NAD(+)-dependent DNA ligase is characterized as a promising drug target in several organisms due to its crucial, indispensable role in DNA replication, recombination and DNA repair. We report here the cloning, expression and purification of NAD(+)-dependent DNA ligase of Wolbachia endosymbiont of B. malayi (wBm-LigA) for its molecular characterization. wBm-LigA has all the domains that are present in nearly all the eubacterial NAD(+)-dependent DNA ligases such as N-terminal adenylation domain, OB fold, helix-hairpin-helix (HhH) and BRCT domain except zinc-binding tetracysteine domain. The purified recombinant protein (683-amino acid) was found to be biochemically active and was present in its native form as revealed by the circular dichroism and fluorescence spectra. The purified recombinant enzyme was able to catalyze intramolecular strand joining on a nicked DNA as well as intermolecular joining of the cohesive ends of BstEII restricted lamda DNA in an in vitro assay. The enzyme was localized in the various life-stages of B. malayi parasites by immunoblotting and high enzyme expression was observed in Wolbachia within B. malayi microfilariae and female adult parasites along the hypodermal chords and in the gravid portion as evident by the confocal microscopy. Ours is the first report on this enzyme of Wolbachia and these findings would assist in validating the antifilarial drug target potential of wBm-LigA in future studies.