Sagartirtha Sarkar

Upregulation of lysyl oxidase and MMPs during cardiac remodeling in human dilated cardiomyopathy

Objective Dilated cardiomyopathy (DCM) represents a large subset of patients with congestive heart failure (HF), and myocardial fibrosis has been shown to be associated with this process. Lysyl oxidase (LOX), a key enzyme, plays a potential role in the biogenesis of connective tissue matrices by catalyzing crosslinks in collagen and elastin. However, the mechanisms involved in the remodeling process during HF are not clearly understood. The present work was aimed to determine the changes in collagen phenotypes, MMPs, TIMPs, and LOX, in DCM and non-failing human hearts. Moreover, the role of TGFβ in the induction of type III collagen in cardiac fibroblast is determined. Method Protein and RNA expression were quantified by Western and RT-PCR analysis; collagen phenotypes were determined by SDS-PAGE. Results Our data demonstrated that in all DCM hearts, the collagen concentration was significantly elevated compared to that of the NF hearts associated with an increase in Type I (18%) and Type III (33%) collagen. The content of MMP-2 and MMP-9 were increased significantly in all DCM hearts compared to NF hearts. Transcriptional level of LOX, TIMP 1, and 2 were significantly upregulated in DCM hearts. In addition, a significant increase in the transcript levels of cytokines, notably IFN, IL-6, TNF-α, and TGF-β superfamily was observed in all DCM hearts. Addition of TGFβ to cardiac fibroblasts caused a dose dependent increase in type III collagen. Conclusion Altogether, our data suggest an alteration of collagen, MMPs, various cytokines and particularly, LOX participates, in part, in the remodeling of the heart leading to cardiac dysfunction and HF.

Rotavirus Nonstructural Protein 1 Suppresses Virus-Induced Cellular Apoptosis To Facilitate Viral Growth by Activating the Cell Survival Pathways during Early Stages of Infection

ABSTRACT Following virus infection, one of the cellular responses to limit the virus spread is induction of apoptosis. In the present study, we report role of rotavirus nonstructural protein 1 (NSP1) in regulating apoptosis by activating prosurvival pathways such as phosphatidylinositol 3-kinase (PI3K)/Akt and NF-κB (nuclear factor κB) during early hours of infections (2 to 8 hpi). The NSP1 mutant strain A5-16 induces weak and transient activation of Akt (protein kinase B) and p65 NF-κB compared to the isogenic wild-type strain A5-13 in MA104 or HT29 cells. The weak NF-κB promoter activity or Akt phosphorylation after A5-16 infection could be complemented in cells transfected with plasmid expressing NSP1 after infection with the rotavirus A5-16 strain. In cells either infected with A5-13 or transfected with pcD-NSP1, coimmunoprecipitation of NSP1 with phosphoinositide 3-kinase (PI3K) was observed, indicating that strong activation of PI3K/Akt could be due to its interaction with NSP1. In addition, after infection with same multiplicity of infection, A5-16 showed reduced number of viral particles compared to the A5-13 strain at the end of the replication cycle. A lower growth rate could be due to weak induction of PI3K/Akt and NF-κB, since the A5-13 strain also showed reduced growth in the presence of PI3K or NF-κB inhibitors. This effect was interferon independent; however, it was partly due to significantly higher caspase-3 activity, poly-ADP ribose polymerase (PARP) cleavage, and apoptosis during earlier stages of infection with the NSP1 mutant. Thus, our data suggest that NSP1 positively supports rotavirus growth by suppression of premature apoptosis for improved virus growth after infection.

Inhibition of Signal Transducer and Activator of Transcription 3 (STAT3) Attenuates Interleukin-6 (IL-6)-induced Collagen Synthesis and Resultant Hypertrophy in Rat Heart

IL-6 has been shown to play a major role in collagen up-regulation process during cardiac hypertrophy, although the precise mechanism is still not known. In this study we have analyzed the mechanism by which IL-6 modulates cardiac hypertrophy. For the in vitro model, IL-6-treated cultured cardiac fibroblasts were used, whereas the in vivo cardiac hypertrophy model was generated by renal artery ligation in adult male Wistar rats (Rattus norvegicus). During induction of hypertrophy, increased phosphorylation of STAT1, STAT3, MAPK, and ERK proteins was observed both in vitro and in vivo. Treatment of fibroblasts with specific inhibitors for STAT1 (fludarabine, 50 μm), STAT3 (S31-201, 10 μm), p38 MAPK (SB203580, 10 μm), and ERK1/2 (U0126, 10 μm) resulted in down-regulation of IL-6-induced phosphorylation of specific proteins; however, only S31-201 and SB203580 inhibited collagen biosynthesis. In ligated rats in vivo, only STAT3 inhibitors resulted in significant decrease in collagen synthesis and hypertrophy markers such as atrial natriuretic factor and β-myosin heavy chain. In addition, decreased heart weight to body weight ratio and improved cardiac function as measured by echocardiography was evident in animals treated with STAT3 inhibitor or siRNA. Compared with IL-6 neutralization, more pronounced down-regulation of collagen synthesis and regression of hypertrophy was observed with STAT3 inhibition, suggesting that STAT3 is the major downstream signaling molecule and a potential therapeutic target for cardiac hypertrophy.

Role of Myocytes in Myocardial Collagen Production

Excessive collagen deposition may cause abnormal stiffness of the heart during hypertrophy and heart failure. The potent vasoconstrictor angiotensin II seems, via an unknown mechanism, to stimulate collagen production. This study describes the in vitro and ex vivo effects of [Sar1]Ang II on collagen production by fibroblasts in culture and in beating, nonworking heart preparations. The effects of [Sar1]Ang II on isolated rat hearts or rat heart fibroblasts were determined by quantifying transcript levels of collagen phenotypes I and III through videodensitometry after Northern blot analysis with specific cDNA probes (collagen [P &agr;2r2] rat &agr;2[I] probe for type I and human skin fibroblast &agr;1[III] probe for type III). When [Sar1]Ang II was added in vitro to neonatal or adult 28-week-old Wistar-Kyoto rat heart fibroblasts, questionable stimulation in the mRNAs of types I and III occurred. In contrast, when 10−8 mol/L [Sar1]Ang II was added to beating, nonworking Wistar-Kyoto rat heart preparation ex vivo, a 1.5- to 2.5-fold stimulation of collagen mRNAs of phenotypes I and III was observed. When neonatal fibroblasts were cocultured with neonatal myocytes in vitro, with 10−10 mol/L [Sar1]Ang II added, there was no stimulation of either phenotype. However, significant stimulation of both collagen transcripts was recorded when 10−10 mol/L [Sar1]Ang II was added to adult fibroblasts cocultured with either neonatal or adult myocytes. Our data suggest that factors produced by myocytes are necessary for upregulation of collagen genes in vitro and demonstrate that fibroblast-myocyte cross-talk is required for Ang II–induced collagen upregulation.

Influence of p53 in the transition of myotrophin-induced cardiac hypertrophy to heart failure

AIMS Cardiac-specific overexpression of myotrophin (myo) protein in transgenic (myo-Tg) mice results in hypertrophy at 4 weeks that progresses to heart failure (HF) by 36 weeks. Gene profiling showed that p53 expression increases as hypertrophy worsens to HF, suggesting that p53 may influence myo-induced HF. We aimed to define how the p53 signalling cascade affects the spectrum of cardiac hypertrophy (CH)/HF. METHODS AND RESULTS Immunoblot analysis showed that in myo-Tg mice (Mus musculus), upregulation of p53 occurs only when hypertrophy transitions to HF (16 weeks onward). To elucidate the role of p53, a double-Tg mouse line (p53(-/-)/myo(+/+)) was developed by crossing myo-Tg mice with p53-null mice. A significant reduction in cardiac mass with improved cardiac function was observed in p53(-/-)/myo(+/+) mice, suggesting that absence of p53 prevents hypertrophy from turning into HF. Analysis via real-time reverse-transcription PCR revealed changes in transcripts of the p53 pathway in p53(-/-)/myo(+/+) mice. Ingenuity Pathway Analysis indicated that cross-talk among several key nodal molecules (e.g. cyclin-dependent kinase inhibitor 1A, caspase-3, nuclear factor kappa-light-chain enhancer of activated B cells etc.) may play a regulatory role in the transition of CH to HF. CONCLUSION Our data provide evidence, for the first time, that the coherence of p53 with myo plays an active role during the transition of CH to HF in a model of HF induced by myo overexpression. Transition from CH to HF can be prevented in the absence of p53 in myo-induced hypertrophy. Therefore, deletion/inhibition of p53 could be a therapeutic strategy to prevent CH from transitioning to HF.

Arjunolic acid, a peroxisome proliferator-activated receptor α agonist, regresses cardiac fibrosis by inhibiting non-canonical TGF-β signaling

Cardiac hypertrophy and associated heart fibrosis remain a major cause of death worldwide. Phytochemicals have gained attention as alternative therapeutics for managing cardiovascular diseases. These include the extract from the plant Terminalia arjuna, which is a popular cardioprotectant and may prevent or slow progression of pathological hypertrophy to heart failure. Here, we investigated the mode of action of a principal bioactive T. arjuna compound, arjunolic acid (AA), in ameliorating hemodynamic load-induced cardiac fibrosis and identified its intracellular target. Our data revealed that AA significantly represses collagen expression and improves cardiac function during hypertrophy. We found that AA binds to and stabilizes the ligand-binding domain of peroxisome proliferator-activated receptor α (PPARα) and increases its expression during cardiac hypertrophy. PPARα knockdown during AA treatment in hypertrophy samples, including angiotensin II-treated adult cardiac fibroblasts and renal artery-ligated rat heart, suggests that AA-driven cardioprotection primarily arises from PPARα agonism. Moreover, AA-induced PPARα up-regulation leads to repression of TGF-β signaling, specifically by inhibiting TGF-β-activated kinase1 (TAK1) phosphorylation. We observed that PPARα directly interacts with TAK1, predominantly via PPARα N-terminal transactivation domain (AF-1) thereby masking the TAK1 kinase domain. The AA-induced PPARα-bound TAK1 level thereby shows inverse correlation with the phosphorylation level of TAK1 and subsequent reduction in p38 MAPK and NF-κBp65 activation, ultimately culminating in amelioration of excess collagen synthesis in cardiac hypertrophy. In conclusion, our findings unravel the mechanism of AA action in regressing hypertrophy-associated cardiac fibrosis by assigning a role of AA as a PPARα agonist that inactivates non-canonical TGF-β signaling.

Role of α -Crystallin B in Regulation of Stress Induced Cardiomyocyte Apoptosis

Cardiovascular disease is the leading cause of death worldwide. Recently emerging evidence suggests that cardiomyocyte apoptosis is one of the major pathogenic factors in heart diseases leading to heart failure. Cardiomyocytes undergo apoptosis in response to a wide variety of cellular stresses including protein folding stress at Endoplasmic reticulum (ER). Stressed myocytes elicit an adaptive response referred as Unfolded Protein Response (UPR) by inducing accumulation of heat shock proteins (HSPs) to mitigate the ER stress. HSPs act as molecular chaperons by assisting correct folding of the aggregated misfolded proteins in ER lumen. α-Crystallin B (CRYAB) is an abundant small HSP that confers protection to cardiomyocytes against various stress stimuli. Recent evidence indicates that CRYAB directly interacts with several components of ER stress and also mitochondrial apoptotic pathway. Based on currently available literature this mini review will focus on how CRYAB confers protection to stressed myocardium thereby emphasizing its function as antiapoptotic molecule. Understanding the interplay between CRYAB and the key components in the apoptotic signaling cascade mediated by ER and mitochondria will help in development of novel therapies for cardiac diseases.

Field populations of native Indian honey bees from pesticide intensive agricultural landscape show signs of impaired olfaction

Little information is available regarding the adverse effects of pesticides on natural honey bee populations. This study highlights the detrimental effects of pesticides on honey bee olfaction through behavioural studies, scanning electron microscopic imaging of antennal sensillae and confocal microscopic studies of honey bee brains for calcium ions on Apis cerana, a native Indian honey bee species. There was a significant decrease in proboscis extension response and biologically active free calcium ions and adverse changes in antennal sensillae in pesticide exposed field honey bee populations compared to morphometrically similar honey bees sampled from low/no pesticide sites. Controlled laboratory experiments corroborated these findings. This study reports for the first time the changes in antennal sensillae, expression of Calpain 1(an important calcium binding protein) and resting state free calcium in brains of honey bees exposed to pesticide stress.

Targeting Insulin-Like Growth Factor-I receptor signaling pathways improve compromised function during cardiac hypertrophy

Insulin-like growth factor-I receptor (IGF-IR) and its signaling pathways play an important role in cell proliferation, growth, differentiation and survival. IGF-axis controls cellular, physiological and metabolic activity of several cell types including cardiomyocytes. IGF-I acts through its signaling receptorIGF-IR to induce physiological or adaptive hypertrophic growth. Various reports also confirm that it protects the heart against pathological insults. Furthermore, deregulation of IGF-I signaling has also been shown to be involved in the development of pathological hypertrophy. Although it would be beneficial to activate the IGF-I–PI3-K pathway to enhance cardiac function during pathological conditions it could be detrimental in case of cancer. Based on currently available literature this mini review will focus on the recent advances of drug and delivery systems that specifically target IGF-I signaling for promoting physiological hypertrophy and improving cardiac function. Abbreviations: Insulin-like growth factor-I (IGF-I); Insulin-like growth factor-I receptor (IGF-IR); Mammalian target of rapamycin (mTOR); 4E binding protein1 (4EBP1); local IGF-1 isoform (mIGF‐1); (PI3K)-AKT/PKB phosphatidylinositol 3-kinase-Akt/Protein Kinase B; Insulin receptor (IR); MicroRNAs (miRs); Ribosomal S6 kinases (S6Ks)

Cardiac tissue targeted delivery of therapeutic payloads successfully regresses cardiac hypertrophy abating bystander effects

A hydrocarbons have attracted concern as a kind of environmental pollutants along with possible health threats. Comamonas testosteroni strain P19 was originally isolated as a biphenyl degrader and capable of degrading several aromatic compounds as a sole source of carbon and energy. Here, we describe the metabolic insight by genetic and physiological analysis indicating how aromatic compounds are metabolized in C. testosteroni strain P19. The strain has a circular chromosome of 5,633,218 bp with a G+C content of 67.65% and one plasmid. The annotation identified gene clusters encoding essential proteins for metabolizing several aromatic hydrocarbons including anthranilate, benzoate, biphenyl, m-hydroxybenzoate, p-hydroxybenzoate, p-methoxybenzoate, phenol, phthalate, protocatechuate, terephthalate etc. Random plasposon mutagenesis confirmed that the predicted gene clusters were essential in the metabolic pathways of the compounds. In addition, we identified novel metabolite on degradation of ferulate which had very similar structure to vanillate. Based on structural assignment, the unknown product was revealed as a dimer of vanillate that was a novel intermediate in ferulate degradation pathway. Subsequently, genomic approach suggested the metabolic network of aromatic compounds in C. testosteroni strain P19.S enterica serovar typhimurium (Salmonella typhimurium) is the most common causative agent of human gastroenteritis after consumption of contaminated seafood. The use of lytic bacteriophages against this pathogen can be a new and promising approach for the prevention of food-contamination and food-borne infection. This study investigated the potential application of the bacteriophage SE-5 during depuration to reduce S. typhimurium in cockles (Cerastoderma edule) at different multiplicity of infection (MOI). Cockles were infected with 106 colony-forming units (CFU)/mL of S. typhimurium in the seawater and each infected group was treated with four different MOI values: 100, 10, 1 and 0.1. Infected cockles were depurated in non-recirculating seawater at 16°C for 12 hour. After S. typhimurium accumulation at 16° C, the initial mean values of bacteria in cockles were 6.20 log CFU/g. Depuration with phages at MOI 0.1 was the best condition to inactivate S. typhimurium in cockles, the concentration was reduced by 1.7 log CFU/g after 2 hours of depuration. Reduction for the other MOI values (MOI=1; MOI=10; MOI=100) was 1.13, 1.21 and 2.10 log CFU/g after 6 hours, 12 hours and 12 hours of treatment respectively. To our knowledge, this is the first report of a depuration trial using bacteriophage in the cockle treatment process. The data of this study indicate clearly that the application of the bacteriophages could reduce significantly the population of S. typhimurium in infected cockles. Moreover, cockles could be maintained alive during the depuration process. Therefore, the application of bacteriophage was effectively proven to be useful for shellfish depuration.S members of the genus Lavandula (Lavenders) are economically important crop plants that produce and store large quantities of essential oils (EO; a mixture of primarily monoterpenes) in glandular trichomes, or oil glands. Because glandular trichomes are specialized for EO production, they strongly express the related structural and regulatory genes. We have generated substantial genomic resources to probe essential oil metabolism in lavenders, and have cloned several key EO biosynthetic genes including lavandulyl diphosphate synthase, caryophyllene, borneol dehydrogenase, cineole synthase, and beta-phellandrene synthase from these plants. We have also developed a set of EST-SSR markers capable of delineating a number of lavender species effectively. In this presentation the author will present a summary of his recent findings.This novel, albeit abstract representation of alphaIntroduction The superfamily of 2OG-dependent dioxygenases, by definition, comprises members from all major taxa, and participates in neutralizing the effects of several forms of abiotic stress, viz., pesticides, hypoxia, and osmotic. The oxidative decarboxylation of 2-oxoglutarate to succinate is coupled with a concomitant substrate hydroxylation and, is in most cases, followed by an additional specialized conversion.W optics has historically been mainly used for imaging applications, light has now become a tool of manipulation and direct interaction with biological samples. It is well known that when light impinges on an object, it exerts a small pressure on it known as optical force. An immediate application of optical forces is found in optical tweezers, which in biology are usually employed as a non-invasive technique to immobilize cellular organisms for imaging purposes. These techniques, including more pioneering ideas such as, e.g., cell optical sorting and in-vivo manipulation, require a precise understanding on how the optical pressure affects biological organisms. This calls for the development of precise numerical methods, whose development will be crucial for applications of the aforementioned techniques. In this respect, ab-initio techniques represent a very important tool that can provide quantitative answers to the problem. By employing a series of massively parallel FDTD simulations, we study how optical forces act on biological matter. As a representative case study, we here consider deformed Red Blood Cells (RBC) illuminated by a monochromatic plane wave. We consider typical deformations arising due to the presence of a disease such as malaria. Realistic parameters for the geometry and the refractive index are then taken from published experiments. In our theoretical campaign, we study the dependence of the optical force on the particle shape and the incident wavelength. We show that optical forces change appreciably with the deformation, with amplitude variation in the pN range for RBCs affected by disease.Background: Epidermal Growth Factor receptor (EGFR) family is a group of four highly conserved trans-membrane tyrosine kinase receptors (EGFR, ErbB2/HER2, ErbB3/HER3 and ErbB4/HER4) involved in fate decision of many pivotal biological processes. Several mutations on EGFR have been associated to number of human cancers making it one of the most sought after target for cancer therapeutics. However despite its therapeutic importance, its full length structural behavior is still illusive. Therefore to utilize and exploit this magic drug target, it is imperative to decipher its structure.Most immunosuppressive agents were initially developed as antibiotics produced by the genus Streptomyces . This investigation was devoted to explore the bioactive metabolite of the Streptomyces variabilis ASU319 extract and testing the purified active compound of this extract as an immunosuppressive agent in rats blood. Elucidation of the chemical structure and optimization of the active compound were studied as well. Antimicrobial activity was conducted using agar-well diffusion and disc diffusion assays. The antimicrobial metabolite was extracted from the fermentation broth by ethyl acetate and purified by TLC and silica gel column chromatography. The pure active compound was then subjected to spectroscopic analyses: 1 H NMR, Elemental analysis, IR and Mass spectra. The active antimicrobial compound was tested as an immunosuppressive agent by injection in the rat blood and the complete blood count (CBC) was determined. The crude extracts of the selected active antagonistic five isolates were tested to prevent the inflammation and proliferation of lymphocytes of the rats blood. The active antimicrobial compound of Streptomyces ASU319 was purified and proven as an immunosuppressive agent. The tested compound decreased each of the neutrophils, lymphocytes and monocytes than the positive control. The compound was of molecular weight 458 g/mol and had given the proposed chemical formula C 24 H 46 O 8 . The most potent bacterial isolate was identified by 16SrRNA sequencing as Streptomyces variabilis ASU319 with accession number [GenBank: KC145278]. These results revealed that Streptomyces variabilis ASU319 is potential microbial for production of active antimicrobial compound that has the ability to decrease the proliferation of the lymphocytes cells in the blood and may be a good immunosuppressive agent.New fluorescent N-(2 and 3-methoxyphenyl) thieno [3, 2-b] pyridin-7-amines were synthesized by C-N coupling of 7-bromothieno [3, 2-b] pyridine with 2 or 3 methoxy anilines and gave very low growth inhibitory (GI50) values when studied in human tumor cell lines but also in non-tumor cells. So, pursuing a future clinical administration of these compounds, they were encapsulated in nanoliposomes due to the toxicity presented in vitro in non-tumor cells. The compounds are reasonably fluorescent in solvents of different polarity exhibiting fluorescence quantum yields between 10% and 60%. Nanoliposomes are technological developments for the encapsulation and delivery of bioactive agents. Because of their biocompatibility and biodegradability along with their size, nanoliposomes have potential applications in a vast range of fields including nanotherapy. Nanoliposomes are able to enhance the performance of bioactive agents by improving their bioavailability in vitro and in vivo stability as well as preventing their unwanted interactions with other molecules. These compounds have been successfully encapsulated in different nanoliposome formulations as revealed by fluorescence emission and fluorescence anisotropy measurements. These results are important for future drug delivery developments using these compounds as antitumor agents.H infections are present worldwide and microorganisms such as viruses, bacteria, fungi and parasites can be responsible for such infections. Fungal infections exhibit low occurrence; however, because they take longer to detect and treat, high rates of morbidity and mortality are known. Thus, fungal infections namely those associated to Aspergillus niger represent actual challenges especially for elderly, neonates or those having an underlying disease or under immunosuppressive drug treatments. Microbial metabolomics has been breaking new ground as a very useful tool in several areas including those related to microbial diagnosis. Microorganisms produce several volatile metabolites, the whole of which can be used as unique chemical fingerprints of each species and possibly of strain. This richness of information holds the promise for diagnosing infections. This research study aims to in-depth study the A. niger exometabolome in order to establish metabolites pattern that can be further exploited for fungal diagnosis. A methodology based on advanced multidimensional gas chromatography (HSSPME/GC×GC-ToFMS) tandem with multivariate analysis was developed. Different growth conditions were assayed. A. niger exometabolome revealed around 500 metabolites distributed over several chemical families being the major ones alcohols, aldehydes, esters, hydrocarbons, ketones and terpenoids. A subset of 44 metabolites defined as the A. niger metabolomic biomarkers pattern was selected and successfully used to distinguish this species from others previously chosen revealing its useful potential.T cell phone technology is an integral part of everyday life and, it is used to chat including voice and picture and in addition has many other applications. Male reproductive system is highly compartmentalized and sensitive biological system that requires the integration of intrinsic and extrinsic factors to normal function, hence we studied effects of GMS (Towers and Mobile radiation) on reproductive hormones (FSH, AMH, LH, Testosterone) and interference of other factors such as smoking, distant and hours near towers, mobile using period, and tumors related with. Our study was applied on 144 samples and was compared with 50 controls. The result showed significant increase in LH & FSH levels while significant decrease in Testosterone & AMH hormone, also there was a significant increase in hormone levels of smoking and nonsmoking persons and distend from tower and a large percentage of tumor disease.M pulegium and Eucalyptus camaldulensis are important plant species with applications in flavouring processed foods. Their essential oils were tested for antifungal activity to explore biocontrol ways vs. fungal rot of apples in storage. Extracts analyses by gas chromatography-mass spectrometry revealed M. pulegium dominated by pulegone and E. camaldulensis, by 1,8 cineole and α pinene. Oils antifungal activity was studied vs. Alternaria alternata and Penicillium expansum. Oil of M. pulegium is thrice most active than E. camaldulensis. Chemical components show synergism. Inhibition suggests food-preservation application. Classification algorithms are proposed based on information entropy and its production. Oils components are classified by numbers of C=C bonds, O atoms and cycles. Classification algorithms are based on information entropy. When applying procedures to moderate-size sets, excessive results appear compatible with data and suffer combinatorial explosion; however, after equipartition conjecture one obtains selection criterion resulting from classification between hierarchical trees. Information entropy permits classifying oils components and agrees with principal component analyses.H metals are common environmental pollutants to the aquatic organisms. Several aquatic species have been used as biomarkers and bio-monitoring subjects for heavy metal pollution. Behavioral changes are sensitive markers of toxicity. The effects of Cd and Mn on the survival, locomotion, attachment and feeding behaviors of the gastropod snail Biomphalaria alexandrina were determined. The 96-h LC50 for Cd and Mn were 0.38 and 156.57 mg/l, respectively. Snails were exposed to sub-lethal concentrations of each metal plus a control for chronic exposure period (16-20 days) and the survival curves were determined. Locomotion and feeding behaviors of snails exposed to Cd and Mn at acute (96 h exposure) and chronic exposure (24 days exposure) intervals were recorded. The survival curves showed that long term exposure of snails to ascending concentrations of Cd and Mn caused a gradual decrease in the survival rate of B. alexandrina in a dose-dependent manner. Compared to control, a significant decrease was recorded in the feeding and locomotion behaviors of exposed snails. The tendency to feed in B. alexandrina was significantly decreased by acute exposure to Cd and completely blocked by Mn exposure. The feeding rate was 4.8±0.68 bites/min. in Cd-exposed snails compared to 16.3±1.7 bites/minute in control. A significant decrease was recorded in the locomotion behavior of exposed snail groups compared to control. The path length of traces was 3351.2±95.3 mm in control compared to 295.3±50 and 1610.5±58.9 mm, respectively for Mn and Cd exposed snails. Thus, sub-lethal metal concentrations affected B. alexandrina behaviors, potentially impacting the snail’s activity and tendency to feed. The present study also demonstrated B. alexandrina as a sensitive bio-indicator and can be used as a model organism to assess heavy metals risk factors for severe toxicity in freshwater ecosystems.1 (ACC) deaminase promotes plant growth by sequestering and cleaving the ethylene precursor ACC to α-ketobutyrate and ammonium. Many plant growth promoting rhizobacteria producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase as a source of nitrogen has an eminent role in plant nutrition. The present work deals with comparative analysis of ACCD producing plant growth-promoting rhizobacteria (PGPR) are Azospirillum lipoferum, Phyllobacterium brassicacearum, Pseudomonas fluorescens, Francisella tularensis subsp. holarctica OSU18 and Bacillus cereus. The sequence and phylogenetic analysis of ACCD producing PGPR species represents the common conserved domain belonging to the tryptophan synthase beta subunit-like PLP-dependent enzymes super family and closely related to each other. The predicted homology models of ACCD of PGPR have similar protein structure with similar folds often share similar function. This analysis represents the evolutionary conservation and same biochemical function of ACCD producing plant growth-promoting rhizobacteria. This analysis is very helpful to understand the biological function of PGPR species.Today, cell phone technology is an integral part of everyday life and, its use to chat including voice and picture in addition to many other applications. Male reproductive system is highly compartmentalized and sensitive biological system that requires the integration of intrinsic and extrinsic factors to normal function, hence we studied effects of GMS (Towers and Mobil radiation)on reproductive hormones (FSH,AMH,LH, Testosterone) and interference of other factors such as smoking ,distant and hours near towers, Mobil using period, and tumors related with .Our study applied on (144) sample compared with (50) control, the result showed significant increase in LH&FSH levels while significant decrease in Testosterone & AMH hormone, also significant increase inhormone levels smoking and non-smoking persons and distend from tower.S steroid hormones regulate multiple female reproductive functions. Cyclooxygenase-2 (COX-2) is an inflammationassociated enzyme to regulate prostaglandin production. Previous studies demonstrated that COX-2-deficient mice failed to ovulate suggesting a vital role of COX-2 in ovulation. Sex steroid hormones were reported to modulate COX-2 expression: Dihydrotestosterone (DHT) was able to inhibit interlukin-1β-induced COX-2 expression in vascular smooth muscle cells while estradiol (E2) was able to promote COX-2 expression in the rat oviduct. In the ovary, the significance and the involving mechanisms of androgens and estrogens in COX-2 regulation remain mostly unclear. The aim of this study was to clarify whether and how sex steroid hormones affect COX-2 expression in rat ovarian granulosa cells. Previous studies suggested that PKC could be activated by FSH or LH in follicular granulosa cells leading to inflammatory-like consequences. Thus, a PKC activator PDD (phorbol-12, 13-didecanoate) was used in this study. It was noted that DHT appeared to attenuate PDDinduced COX-2 protein, mRNA expression and promoter activity; However, E2 was able to enhance PDD induced COX-2 protein, mRNA expression and promoter activity. In addition, the PDD-mediated PGE2 production was also impacted by DHT and E2. The PDD-mediated COX-2 expression was inhibited by parthenolide (NF-κB inhibitor) but was enhanced by SP600125 (JNK inhibitor) or wortmannin (PI3K inhibitor). Thus, DHT and E2 may affect PKC-mediated inflammation in ovarian granulosa cells by acting through these signaling players.D to the natural complementary properties in DNA sequences based on double helix pair exploring longer DNA pieces and its reverse fragment in complementary symmetry with a distance play an important role in modern genetic analysis. In this talk, the substring complementary string matching technique is used to detect and analyze the structure of DNA motif. We focused on the relationship between fragment and its reverse fragment in complementary symmetry and the possible structure of palindromic sequences, hairpin prediction. Statistical measurements are used to process some selected human genome data and all possible corresponding fragments are identified. We convert the identified data into visualized figures to illustrate the visible analysis results of quantities of complex DNA sequences as extracted information.