Novel Oleanolic Acid-Phtalimidines Tethered 1,2,3 Triazole Hybrids as Promising Antibacterial Agents: Design, Synthesis, In Vitro Experiments and In Silico Docking Studies.

As part of the valorization of agricultural waste into bioactive compounds, a series of structurally novel oleanolic acid ((3ß-hydroxyolean-12-en-28-oic acid, OA-1)-phtalimidines (isoindolinones) conjugates 18a-u bearing 1,2,3-triazole moieties were designed and synthesized by treating an azide 4 previously prepared from OA-1 isolated from olive pomace (Olea europaea L.) with a wide range of propargylated phtalimidines using the Cu(I)-catalyzed click chemistry approach. OA-1 and its newly prepared analogues, 18a-u, were screened in vitro for their antibacterial activity against two Gram-positive bacteria, Staphylococcus aureus and Listeria monocytogenes, and two Gram-negative bacteria, Salmonella thyphimurium and Pseudomonas aeruginosa. Attractive results were obtained, notably against L. monocytogenes. Compounds 18d, 18g, and 18h exhibited the highest antibacterial activity when compared with OA-1 and other compounds in the series against tested pathogenic bacterial strains. A molecular docking study was performed to explore the binding mode of the most active derivatives into the active site of the ABC substrate-binding protein Lmo0181 from L. monocytogenes. Results showed the importance of both hydrogen bonding and hydrophobic interactions with the target protein and are in favor of the experimental data.

Potential functions of hsa-miR-155-5p and core genes in chronic myeloid leukemia and emerging role in human cancer: A joint bioinformatics analysis.

Considering the critical roles of hsa-miR-155-5p participated in hematopoietic system, this study aims to clarify the possible pathogenesis of chronic myeloid leukemia (CML) induced by hsa-miR-155-5p.Three different strategies were employed, namely a network-based pipeline, a survival analysis and genetic screening method, and a simulation modeling approach, to assess the oncogenic role of hsa-miR-155-5p in CML. We identified new potential roles of hsa-miR-155-5p in CML, involving the BCR/ABL-mediated leukemogenesis through MAPK signaling. Several promising targets including E2F2, KRAS and FLI1 were screened as candidate diagnostic marker genes. The survival analysis revealed that mRNA expression of E2F2, KRAS and FLI1 was negatively correlated with hsa-miR-155-5p and these targets were significantly associated with poor overall survival. Furthermore, an overlap between CML-related genes and hsa-miR-155-5p target genes was revealed using competing endogenous RNA (ceRNA) networks analysis. Taken together, our results reveal the dynamic regulatory aspect of hsa-miR-155-5p as potential player in CML pathogenesis.

Comprehensive in-silico analysis of damage associated SNPs in hOCT1 affecting Imatinib response in chronic myeloid leukemia.

Non-synonymous single nucleotide polymorphisms (nsSNPs) in hOCT1 (encoded by SLC22A1 gene) are expected to affect Imatinib uptake in chronic myeloid leukemia (CML). In this study, sequence homology-based genetic analysis of a set of 270 coding SNPs identified 18 nsSNPs to be putatively damaging/deleterious using eight different algorithms. Subsequently, based on conservation of amino acid residues, stability analysis, posttranscriptional modifications, and solvent accessibility analysis, the possible structural-functional relationship was established for high-confidence nsSNPs. Furthermore, based on the modeling results, some dissimilarities of mutant type amino acids from wild-type amino acids such as size, charge, interaction and hydrophobicity were revealed. Three highly deleterious mutations consisting of P283L, G401S and R402G in SLC22A1 gene that may alter the protein structure, function and stability were identified. These results provide a filtered data to explore the effect of uncharacterized nsSNP and find their association with Imatinib resistance in CML.

Atypical Salmonella Typhimurium persistence in the pacific oyster, Crassostrea gigas, and its effect on the variation of gene expression involved in the oyster's immune system.

Salmonella is one of the most important pathogens involved in food intoxication outbreaks, and in many cases, the intoxication has been linked to shellfish which is typically consumed raw. While much is understood about the interactions between Salmonella and vertebrates, much less is known about its relationships with invertebrates, which could be an overlooked and important aspect to better understand the Salmonella interaction with its diversified hosts. The aim of this study was to investigate the effect of preadaptation in seawater microcosms during 12 months on Salmonella Typhimurium by determining its survival capacity within this mollusk over a period of 30 days. The results showed that the stressed bacteria are able to survive in this mollusk at a higher concentration even after thirty days of infection compared to bacteria in the normal state. In order to minimize the effect of an experimental device for one month on the survival of Salmonella, we carried out an in vitro study to determine the number of viable Salmonella in the hemocytes of oysters. Interestingly, we evaluated the effect of the antibacterial activity of different extracts of C. gigas using the solvents (Methanol, Ethanol and acetic acid) specifically against stressed and unstressed Salmonella. Furthermore, we compared the expression of three genes in the oyster Cg-big-def1, timp and sod in response to experimental infections of this mollusk with Vibrio splendidus kb133 and S. Typhimurium LT2DT104 in normal and stressed states. These findings are very important to contribute to explaining several questions about the persistence of S. Typhimurium for a long time in C. gigas and the host's immune response to this microorganism which is considered to be non-virulent for molluscs.

Use of extracellular polysaccharides, secreted by Lactobacillus plantarum and Bacillus spp., as reducing indole production agents to control biofilm formation and efflux pumps inhibitor in Escherichia coli.

The overuse of antibiotics and biofilm formation ability has led to the emergence of bacterial resistant strains. The combined use of several antibiotics has been found as an efficient strategy to overcome this resistance. In this study, two exopolysaccharides (EPS) obtained from Lactobacillus plantarum (EPS-Lp) and Bacillus spp. (EPS-B), isolated from a traditional Tunisian food "ricotta cheese" and hypersaline environment respectively, were used to counteract the biofilm formation and efflux pumps activities in Escherichia coli ATCC35218. The obtained results revealed that the tested EPSs can be effective against E. coli at a concentration > 1 mg/ml and were able to modulate biofilm formation by 50%. Moreover, at a concentration of 512 µg/ml, the tested EPSs inhibit the EtBr efflux in the tested bacteria and no significant difference was shown compared to cells treated with reserpine (P > 0.05). The positive effect of the tested EPSs may be due to the decrease of Indole production level proposed as a signal involved in quorum sensing and through the significant reduction of the hydrophobicity percentage between the treated and untreated cells. Overall, EPS-Lp and EPS-B, when used at appropriate concentration, may inhibit biofilm formation and reduce efflux pumps implicated in bacterial adhesion and antimicrobial resistance. These results make them an interesting candidate in the design of a new strategies to control bacterial biofilm-associated infections.

Motility, biofilm formation, apoptotic effect and virulence gene expression of atypical Salmonella Typhimurium outside and inside Caco-2 cells.

Disease outbreaks related to waterborne pathogen contamination throughout the world as well as challenges that lie ahead for addressing persistent infection are of renewed interest. In this research, we studied the effects of prolonged exposure of Salmonella enterica serovar Typhimurium to the cues encountered in the extracellular environment particularly in seawater microcosm on bacterial virulence and subsequent infection in Caco-2 cells. Our data show a significant difference in biofilm formation, swimming and swarming motilities between normal and stressed cells of S. Typhimurium under differing NaCl conditions (P < 0.05). Interestingly, adhesion, invasion and apoptotic activity to Caco-2 epithelial cells were determined during infection with normal and stressed Salmonella. Furthermore, we compared the expression of SPI-1 virulence genes (sopA, sopB, sopD, sopE2 and hilA) of normal and stressed S. Typhimurium in response to salt conditions encountered in the extracellular environment in LB broth and after epithelial cell exposure. The interest of the present study is due to the fact that to investigate the bacterial survival strategies during its movement from the natural surroundings to the host cell is fundamental to our understanding of the infection process during the host-pathogen interactions.

Extracellular polysaccharide derived from potential probiotic strain with antioxidant and antibacterial activities as a prebiotic agent to control pathogenic bacterial biofilm formation.

Because of their functional diversity, bioactive compounds are becoming a new biocontrol agent to limit biofilm formation by pathogens. In this study, the physico-chemical characterization of an exopolysaccharide (EPS) isolated from Lactobacillus plantarum (EPLB) was characterized and its in vitro effect on biofilm formation was studied. The EPS had a molecular weight of 36 kDa and polydispersity index estimated to be 1.2. The tested EPLB had an antibacterial activity, with a Minimal Inhibition Concentration (MIC) values ranging between 1 mg/ml and 10 mg/ml, displayed an antibiofilm effect concentration dependent on Gram positive and negative strains. Among the pathogenic strains, 2 out of 4 appeared to be more than 50% inhibited in their biofilm development by the EPS. The antibiofilm activity can be due to the ability of the EPS to influence the function of biological membranes like hydrophobicity that decreased (P < 0.05) when the EPS was used at a concentration of 512 µg/ml. This EPS without cytotoxic effect, showed an antioxidant effect on the quenching of DPPH radicals and the inhibition of lipid peroxidation with a percentage of 64% and 66%, respectively. Taken together these biological properties, EPLB can be considered as a potential prebiotic agent in the design of new therapeutic strategies for bacterial biofilm-associated infections.

Adhesion, invasion, cytotoxic effect and cytokine production in response to atypical Salmonella Typhimurium infection.

Pathogenic bacteria such as Salmonella have the ability to respond to a wide variety of environmental stimuli. These responses allow them to survive and withstand insults both of an external location as well as within the host. The aim of this study was to investigate the effect of preadaptation in stressful conditions encountered in seawater microcosms for different periods of time on Salmonella Typhimurium survival, antibiotic susceptibility and interactions with Caco-2 cells. These results showed that the number of bacterial cells depends from the periods of stress in culture medium, highlighting the importance of using the right culture medium for the enumeration of stressed bacteria. The antibiotic resistance of starved cells was modified and their exposure to stressful conditions in seawater during 12 months significantly increased adhesion, invasion and cytotoxic activities on Caco-2 cells. Moreover, cellular cytokines IL-6 and IL-8 secretions were up-regulated. Present results seem to suggest that the preadaptation of S. Typhimurium in seawater microcosms affect the cultural characters by the appearance of the atypical cells that may play a critical role in the intestinal infection and in the systemic spread of the disease. These findings are very important to understand bacterial responses to changing conditions and explain the persistence of these atypical in eukaryotic cells.

Structural impact, ligand-protein interactions, and molecular phenotypic effects of TGF-ß1 gene variants: In silico analysis with implications for idiopathic pulmonary fibrosis.

BACKGROUND: Idiopathic Pulmonary Fibrosis (IPF) is a chronic interstitial lung disease resulting in progressively deteriorating lung function. Transforming growth factor-ß1 (TGF-ß1) belongs to the TGF superfamily and exerts a profibrotic role in promoting lung fibrosis by facilitating fibroblast infiltration and activity, extracellular matrix deposition, and inhibition of collagen breakdown, thus promoting tissue remodelling and IPF. MATERIALS AND METHODS: We evaluated the link between pathogenic TGF-ß1 SNPs and IPF pathogenesis and the structure-activity functional consequences of those SNPs on the TGF-ß1 protein. Several computational algorithms were merged to address the functional consequences of TGF-ß1 gene mutations to protein stability, putative post-translational modification sites, ligand-protein interactions, and molecular phenotypic effects. These included FATHMM, POLYPHEN2, PROVEAN, and SIFT tools (identifying deleterious nsSNPs in the TGF-ß1 gene), along with Pmut, PhD-SNP, SNAP, MutPred and the related TMHMM, MARCOIL, and DisProt algorithms (predicting structural disorders). INPS-MD was also used to evaluate the mutation-induced TGF-ß1 protein's stability and MODPRED for recognition of post-translational TGF-ß1 modification. RESULTS: In total, 14 major pathogenic variants markedly impact the destabilization of the TGF-ß1 protein, with most of these high-risk mutations associated with decreased stability of the TGF-ß1 protein as per the I-Mutant, MUpro, and INPS-MD tools. R205W, R185W, R180Q, D86Y, and I300T variants were proposed to participate in the post-translational modifications, thus affecting affect protein-ligand interactions. Furthermore, at-risk genetic variants appear to target conserved regions in the alpha helices, random coils, and extracellular loops, resulting in a varied composition of amino acids, charge, hydrophobicity, and spatial architecture. CONCLUSIONS: This study manuscript comprehensively analyzes gene variants within the TGF-ß1 gene, offering novel insights into their structural and functional implications in interacting with target sites. This study is significant for the development of targeted therapeutic strategies and personalized treatment approaches for patients with inflammatory lung diseases such as IPF.

Use of mixture design to construct a consortium of date palm (Phoenix dactylifera L.) fruit extract and potentially probiotic Bacillus strain to confer protection against vibriosis in Artemia culture.

BACKGROUND: To find a biological control treatment applicable to shrimp hatcheries, the influence of the dietary administration of a consortium of two date palm (Phoenix dactylifera L.) fruit extracts and potentially probiotic Bacillus strain on Artemia culture was studied. Challenge tests were performed with virulent Vibrio alginolyticus to assess the protective effects of Deglet Nour and Degla fruit extracts alone or in conjunction with potentially probiotic Bacillus strain. RESULTS: Under the conducted bioassays, where Deglet Nour and Degla fruit extracts were used singly or in association with potentially probiotic Bacillus strain, the survival and growth rates of Artemia nauplii were improved. Nauplii protection against virulent Vibrio strain was highest using a feed comprising 2.7% Deglet Nour/97.3% Deglat extract and no Bacillus bacteria. CONCLUSION: The present findings provide evidence that the use of palm fruit extracts singly or in conjunction with potentially probiotic Bacillus strain has tremendous potential as a biocontrol treatment for Artemia culture, particularly with regard to improving culture conditions and the ability to inhibit bacterial pathogens under in vivo conditions.

Development of Nanotechnology-Based Drug Delivery Systems for Controlling Clinical Multidrug-Resistant Staphylococcus aureus and Escherichia coli Associated with Aerobic Vaginitis.

The growing prevalence of resistance to antibiotics potentially makes Escherichia coli and Staphylococcus aureus serious pathogens, necessitating the development of new antimicrobial agents. We extracted crude biosurfactants from a potential probiotic Bacillus spp. to control pathogenic bacteria associated with aerobic vaginal infection. Using nanotechnology formulations, we developed nanoemulsions based on biosurfactants at different concentrations (1% and 3.33%). The results showed that these nanoemulsions were stable, with a weighted index of 0.3, and demonstrated broad-spectrum antibacterial activity against Escherichia coli and Staphylococcus aureus, with MICs ranging between 1.25 and 4 mg/mL. Additionally, the nanoemulsions exhibited interesting antibiofilm effects. All strains became more sensitive to the antibiotics to which they were resistant because of various biosurfactant formulations combined with antibiotics. Lower concentrations of BNE1% and 3.33% were still more efficient than the crude biosurfactants. Our findings demonstrated that the biosurfactant had a strong antibiofilm effect against all tested pathogens. This antibacterial effect can be explained by their ability to alter cell physiology such as cell hydrophobicity and membrane disintegration. Thus, we can conclude that the use of nanotechnology formulations has improved this effect, and the nanoemulsions developed in this study can be used as a potential anti-infectious therapy against multidrug-resistant bacterial strains of clinical origin.

Methicillin-Resistant Staphylococcus aurous: Epidemiology, Transmission and New Alternative Therapies: A Narrative Review.

Over the last decade, we were facing medical struggle by the emergence of multi-resistant bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). MRSA infections are still causing a growing global concern due to the rapid adaptive multidrug resistance to conventional antibiotics in human, community and veterinary medicine. Here we provide an overview about MRSA epidemiology, transmission and alternative potential treatments particularly new discovered phytochemicals with biological activity. In this narrative review, bibliographic data was collected from literature search databases: Google Scholar, web of science and PubMed/MEDLINE during recent years (2016 to 2021). MRSA is responsible of wide spectrum life threatening infections such us septicemia, endocarditis, and wound infections. It has epidemic potential in hospitals, that is responsible of most nosocomial infections leading to mortality and constitute a real burden for the healthcare systems. Effective preventive strategies for management of MRSA are highly required moreover, the identification and development of novel drugs or active biomolecules through phytochemicals are time challenging to face new resistant strains.

Impact of the Extraction Method on the Chemical Composition and Antioxidant Potency of Rosmarinus officinalis L. Extracts.

Rosmarinus officinalis L. is a dietary source that produces polyphenols as secondary metabolites. These natural compounds with potent antioxidant abilities are increasingly recommended as a supplement to inhibit oxidative stress. In the current work, we evaluated the impact of the extraction method on the chemical composition of R. officinalis extract, especially on the content of carnosic (CA) and rosmarinic (RA) acids using UPLC-MS-DAD as well as on their antioxidant potency. Four extracts of Tunisian rosemary were obtained from non-conventional extraction techniques:ultrasound-assisted extraction (UAE),supercritical extraction (SFE) and UAE and SFE combined ((UAE-SFE(I), UAE-SFE(II)). The UAE exhibited the best total phenolic compounds (i.e., 85.27 mg GAEg-1), the highest content of CAand RA and the strongest antioxidant abilities (i.e., IC50 = 0.13 mg/mL and EC50 = 0.93 mg/mL for DPPH scavenging test and iron reducing power ability assay). The evaluation of antioxidant activity of UAE inhuman skin fibroblast (HS-68) cell line was carried out after the induction of oxidative stress. The results determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed a strong protective effect against H2O2oxidative stress induction in cells pretreated with UAE. The obtained results allow us to give new insight about the effect of the extraction method on the chemical composition and biological activities of the extract and the importance of the choice of the most appropriate processing technique to prepare rosemary extract with a high antioxidant potency and protective effect against oxidative stress.

Control of Staphylococcus aureus methicillin resistant isolated from auricular infections using aqueous and methanolic extracts of Ephedra alata.

In the current study the potential use of aqueous and methanolic extracts of Ephedra alata aerial parts as biological control agent against pathogenic bacteria and especially Staphylococcus aureus methicillin resistant isolated from auricular infections was evaluated. Chemical tests and spectrophotometric methods were used for screening and quantification of phytochemicals. The assessment of the antioxidant activity was accomplished by DPPH and ABTS radicals scavenging assays. Extracts were evaluated for their antibacterial efficacy by diffusion and microdilution methods. Biofilm inhibition was tested using XTT assay and the cytotoxicity of extracts was carried out on Vero cell line. The GC-FID analysis revealed that E. alata was rich in unsatured fatty acids. In addition, the aqueous extract had the highest flavonoid and protein contents (30.82 mg QE /g dry extract and 98.92 mg BSAE/g dry extract respectively). However, the methanolic extract had the highest phenolic, sugars and tannins. The antioxidant activity demonstrated that the aqueous extract exhibited the strong potency (IC50 ranged between 0.001 and 0.002 mg/mL). Both extracts displayed antimicrobial activity on Gram negative and positive strains. They were effective against S. aureus isolated from auricular infections. The tested extracts were able to inhibit biofilm formation with concentration-dependent manner. Moreover, no cytotoxic effect on Vero cells line was demonstrated for the extracts. Overall, our findings highlight the potential use of E. alata extract as a novel source of bioactive molecules with antioxidant, antibacterial and antiobiofilm effects for the control of infectious disease especially those associated to S. aureus methicillin resistant.

Microbial Diversity and Pathogenic Properties of Microbiota Associated with Aerobic Vaginitis in Women with Recurrent Pregnancy Loss.

Recurrent pregnancy loss (RPL) is a major reproductive problem that affects approximately 5% of couples. The objective of this study was to assess vaginal flora dysbiosis in women suffering from unexplained RPL and to investigate the pathogenic properties of the microbiota associated with aerobic vaginitis (AV). The study included one hundred fifteen women, 65 with RPL and 50 controls. The diversity of vaginal microbiota isolated was evaluated by molecular sequencing. Then, pathogenic factors, such as acid-resistance, antibiotics susceptibility, and biofilm formation were evaluated. The prevalence of AV was five-fold higher in the RPL group than in the controls (64.6% vs. 12.0%). The most prevalent isolates in the case group were Enterococcus spp. (52%) and Staphylococcus spp. (26%). All bacterial strains tolerate low pH. The prevalence of multidrug resistance (MDR) among all bacteria was 47.7%. Of all strains, 91.0% were biofilm producers. The presence of MDR was found to be related to biofilm formation. The results provide evidence supporting an increased presence of dysbiosis of the vaginal flora, especially AV, in women with RPL in Tunisia. The viability of the AV-associated bacteria and their persistence in the genitals may be due to their ability to resist low pH and to produce a biofilm.

Contribution of ADIPOQ Variants to the Genetic Susceptibility of Recurrent Pregnancy Loss.

Adiponectin is a hormone implicated in regulating energy, lipid, and glucose metabolism and is encoded by the ADIPOQ gene. ADIPOQ variants can regulate the circulating levels of adiponectin. Irregular adiponectin concentrations have been associated with numerous reproductive diseases including recurrent pregnancy loss (RPL). The main objective of this study was to determine whether the 14 selected polymorphisms of the ADIPOQ gene are linked with RPL. The retrospective case-control study comprised a total of 332 women with RPL, adjusted as more than three consecutive abortions of unknown etiology, and 286 healthy controls. They were genotyped for the ADIPOQ variants using allele exclusion method on real-time PCR. Significantly higher rs1501299 minor allele frequencies (MAF) and lower rs2241767 and rs2241766 MAF were seen among RPL women, thereby assigning disease susceptibility and protective aspect to the mentioned variants, respectively. Different associations of ADIPOQ genotypes with RPL were noticed according to the genetic model exploited: rs1501299 and rs2241767 were significantly linked with RPL under the three models, while rs17366568 and rs2241766 were associated with RPL under codominant and dominant models, and rs7649121 was related to RPL under the dominant and recessive models. rs4632532 was linked according to the recessive model only. Based on LD pattern, 2-haplotype blocks were specified. Reduced frequency of AGG and GAGG and increased frequency of TAAG were noted in cases, compared with controls, hence indicating these haplotypes as RPL-protective and RPL-susceptible, respectively. These results support a significant role of ADIPOQ as an RPL candidate locus.

Association of genetic variants in Estrogen receptor (ESR)1 and ESR2 with susceptibility to recurrent pregnancy loss in Tunisian women: A case control study.

Estrogen receptor (ER) signaling is key regulator for maintaining successful pregnancy. Several research suggested that genetic variation in ER genes (ESR)1 and ESR2 is associated with the susceptibility to unexplained recurrent pregnancy loss (RPL), often with inconclusive results. In this study, we investigate the relationship between ESR1 and ESR2 polymorphisms and idiopathic RPL. A total of 444 patients with RPL, defined as three or more consecutive pregnancy losses of unknown etiology, and 446 control women were recruited to the study and their genotypes for ESR1-rs2234693, ESR1-rs3020314, and ESR2-rs928554 variants were determined using allelic exclusion method on real-time polymerase chain reaction. Minor allele frequencies (MAF) of tagging SNPs ESR1 rs2234693 and rs3020314, and ESR2 rs928554 were not significantly different between RPL cases and control women. Considerable higher frequencies of homozygous (2/2) ESR1 rs2234693 genotype carriers were seen between patients vs. control women, which maintained after controlling for age, body mass index (BMI), and menarche. ESR1 haplotype analysis demonstrated two common haplotype (rs2234693-rs3020314) with no linkage disequilibrium between both polymorphisms, and no 2-locus haplotype linked with RPL risk was revealed. The present study confirmed a significant association of specific ESR1 variant (rs2234693) with an increased risk of RPL, further supporting a role for ESR1 as an important candidate locus inducing RPL.

Dietary administration effects of exopolysaccharide from potential probiotic strains on immune and antioxidant status and nutritional value of European sea bass (Dicentrarchus labrax L.).

The use of biological immunostimulants is considered a valuable practice to improve culture conditions in aquaculture sector that may help to increase production and maintain healthy environment. We undertook this study in order to evaluate the potential effect of the administration of two exopolysaccharides (EPS) "EPLB" and "EPB" derived from potential probiotic strains on immune and antioxidant status of European sea bass (Dicentrarchus labrax L.) larvae. In order to find out if the EPSs have an effect on the biochemical composition during the trial period, the nutritional value has been evaluated. The results revealed that expression levels of immune-relevant genes (infg, Il1b, Il8, Il6 and tcr-ß) in the gut and head kidney and the scavenging enzymes (cat, sod, gr) genes in the liver were modulated. In fact, the dietary supplementation with the tested EPSs, significantly enhances the expression of immune-associated genes in the head-kidney, particularly infg and tcrß, as well as catalase gene in liver. During the period of study, EPSs administration did not affect the fatty acid profiles of larvae, which is balanced. This is confirmed by the Docosahexaenoic acid / Eicosapentaenoic acid ratio and demonstrates that EPLB and EPB can be administrated without any negative effect on biochemical composition of European sea bass. The present findings provided evidence that the tested EPSs with antibacterial and antioxidant activities can enhance immune response without negative effect on the biochemical composition. The used EPSs can be considered as a good source of natural functional aquafeed ingredients for European sea bass.

Links between SNPs in TLR-2 and TLR-4 and idiopathic recurrent pregnancy loss.

Background: Recurrent pregnancy loss is a serious complication of pregnancy and failure of the innate immune system, one part of which are toll-like receptors (TLRs). We hypothesised links between variants of TLR-2 and TLR-4 with recurrent pregnancy loss.Subjects and methods: We recruited 335 women with recurrent pregnancy loss, defined as ≥3 consecutive spontaneous miscarriage of unknown aetiology, and 331 age-matched control women. TLR-2 rs1898830 and rs4696483 and TLR-4 rs2770150, rs1554973 and rs7856729 genotyping were performed by allelic exclusion method (real-time PCR).Result: Of the five tested TLR-2 and TLR-4 tag-SNPs, minor allele frequency of TLR-2 rs1898830 was significantly more frequent in recurrent pregnancy loss patients than in controls. Significantly higher frequencies of homozygous (2/2) TLR-2 rs1898830 (14.1% vs. 8.9%) genotype carriers were seen between recurrent pregnancy loss cases and control women. Haploview analysis identified 1-locus TLR-2 haplotype (GC) that was positively associated with recurrent pregnancy loss. No TLR-4 haplotypes associated with altered recurrent pregnancy loss risk were identified.Conclusion: These findings confirm positive associations of TLR-2 rs1898830 with recurrent pregnancy loss, further supporting a role for TLR signalling in defining pregnancy outcome.

Survival and retention of the probiotic properties of Bacillus sp. strains under marine stress starvation conditions and their potential use as a probiotic in Artemia culture.

The probiotic properties of Bacillus strains isolated from Artemia culture and the effect of marine stress on viability and survival were investigated, as well as the changes occurring in their properties. Analyses showed that these bacteria corresponded to the genus Bacillus sp. Antagonism and adherence assays revealed that Bacillus strains have an inhibitory effect against tested pathogenic bacteria and are fairly adherent. Normal and starved cells showed different enzymatic profiles. Challenge tests performed with Artemia larvae provided evidence that the tested Bacillus strains were neither pathogenic nor toxic to the host and conferred protection for Artemia culture against pathogens. The tested strains maintained their viability and their probiotic properties during the period of study. The results suggest that the tested strains have suffered changes allowing them to survive in seawater in the absence of nutrients and outside their natural host, identifying them as potential probiotic candidates for Artemia culture.