Hsp70-2 gene polymorphism: susceptibility implication in Tunisian patients with coronary artery disease

ABSTRACT: Coronary artery disease (CAD) is a multifactorial disease where genetic and environmental factors interact in complex ways to cause the disease. Heat shock protein genes are involved in the progress of CAD. This implies that genetic variants of Hsp70-2 genes might contribute to the development of the disease. Aim of study The aim of this study was to characterize statistical correlation of linkage between lipid profiles, polymorphism PstI site of Hsp70-2 gene and coronary artery diseases. Patients and methods This study was carried out on Tunisian patients with CAD recruited from Hospital of Fattouma Bourguiba of Monastir-Tunisia. Polymerase chain reaction and restriction enzymes were used to determine the genotypic distributions in 252 unrelated patients and 151 healthy control subjects. Further, ApoA-I and ApoB as well as the serum total of cholesterol, HDL, triglyceride, and hs-CRP levels were measured. RESULTS: We showed a decreased level of ApoA-I, whereas the levels of each of ApoB and hs-CRP were increased in patients with CAD compared with control group. In addition our studies of a polymorphic PstI site of Hsp70-2 gene lying in the coding region at position 1267 of the Hsp70-2 gene have revealed that the frequency of P1/P2 heterozygote was 0.484 in patient group compared with control group (0.476, p = 0.046). Whereas, the frequency of the P2/P2 homozygote was 0.190 in patient group and only 0.099 in controls (P = 0.006). These results indicate that the odds ratio of CAD associated with the Hsp70-2 polymorphism is confined the P2/P2 homozygotes (OR 2.498; P = 0.006). CONCLUSION: Taken together, our results indicate that the high frequency of P2/P2 genotype is associated with elevated levels of biochemical parameters (LDL cholesterol, hs-CRP) in Tunisian patient group. The Hsp70-2 polymorphism has susceptibly implication in CAD. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1118340895703689.     

Rapid investigation of French sourdough microbiota by restriction fragment length polymorphism of the 16S-23S rRNA gene intergenic spacer region

The aim of the present research is to identify rapidly the lactic acid bacteria (LAB) microflora of four natural French sourdoughs (GO, BF, VB and RF), applying a biphasic (restriction length polymorphism (RFLP) and sequencing) approach for bacterial identification. For this purpose, a database with the RFLP patterns of 30 lactobacilli type strains was created. So-developed ISR-RFLP algorithm was further applied for the differentiation and identification of 134 sourdough isolates. The 16S-23S rDNA intergenic spacer region was amplified by primers t^Ala and 23S/10, and then digested by HindIII, HinfI and α-TaqI enzymes. Nucleotide sequences of the cloned 16S-23S intergenic spacer region (ISR) were determined by the dideoxynucleotide chain termination method. The T7Prom and M13rev primers flanking the multiple cloning site of pCR2.1 DNA were used to sequence both DNA strands. The RFLP profile obtained upon digestion with HindIII, HinfI and α-TaqI enzymes can be used to discriminate Lactobacillus sanfranciscensis (66%), Lactobacillus panis (17%), Lactobacillus nantensis (11%) and Lactobacillus hammesii(6%) in sourdough GO, Lactobacillus sanfranciscensis (80%), Lactobacillus spicheri (14%) and Lactobacillus pontis(6%) in sourdoughs BF. In sourdoughs VB, which differed in the process temperature, we can differentiate Lactobacillus sanfranciscensis (89%) and Leuconostoc mesenteroidessubsp. mesenteroides (11%). Lactobacillus frumenti(47%), Lactobacillus hammesii (8%), and Lactobacillus paralimentarius (45%) were differentiated in sourdough RF.     

The effect of training at the same time-of-day on the diurnal variations of technical ability and swimming performance

The aim of this study was to examine the effects of training at the same time of day on diurnal variations of technical ability and swimming performance, to provide some recommendations with regard to adjusting training hours in accord with the time of day of competitive events. Eighteen participants volunteered for this study, and these were randomly assigned to either a morning training group (MTG, who trained only between 07:00 and 08:00 h, n = 6), an evening training group (ETG, who trained only between 17:00 and 18:00 h, n = 6), or a control group (CG, did not train but participated in all tests, n = 6). Swimming performance and technical ability – (i) stroke parameters: swim velocity (V), stroke rate (SR), and stroke length (SL); and (ii) motor organization: arm stroke phases and arm coordination (Idc) – were recorded 2 weeks before and 2 weeks after an 8-week regular training period. For all participants, the morning and evening tests were scheduled at the same time of day as the morning and evening training sessions. After training, the major finding of this study was that both ETG and the CG showed significantly lower P, V, SR, phase (B), phase (C), and Idc values in the morning than in the evening. However, P, V, SR, phase (B), phase (C), and Idc of the MTG measured at 07:00 and 17:00 h did not differ. Thus, training at a specific time of day increased performance in MTG at this time and modified the diurnal variation of swim performance. This study indicates that training at a specific time of day can result in marked changes in both swimming performance and technical aspects of swimming. Furthermore, training in the morning improved morning swimming performance and its components, and the amplitude of the morning–evening difference decreased. Training in the evening improved swimming performance and its components more in the evening than the morning, and the amplitude of the morning–evening difference increased.     

Diurnal variations of plasma homocysteine, total antioxidant status, and biological markers of muscle injury during repeated sprint: effect on performance and muscle fatigue--a pilot study

The aim of this study was (i) to evaluate whether homocysteine (Hcy), total antioxidant status (TAS), and biological markers of muscle injury would be affected by time of day (TOD) in football players and (ii) to establish a relationship between diurnal variation of these biomarkers and the daytime rhythm of power and muscle fatigue during repeated sprint ability (RSA) exercise. In counterbalanced order, 12 football (soccer) players performed an RSA test (5 x[6 s of maximal cycling sprint + 24 s of rest]) on two different occasions: 07:00-08:30 h and 17:00-18:30 h. Fasting blood samples were collected from a forearm vein before and 3-5 min after each RSA test. Core temperature, rating of perceived exertion, and performances (i.e., Sprint 1, Sprint 2, and power decrease) during the RSA test were significantly higher at 17:00 than 07:00 h (p < .001, p < .05, and p < .05, respectively). The results also showed significant diurnal variation of resting Hcy levels and all biological markers of muscle injury with acrophases (peak times) observed at 17:00 h. These fluctuations persisted after the RSA test. However, biomarkers of antioxidant status' resting levels (i.e., total antioxidant status, uric acid, and total bilirubin) were higher in the morning. This TOD effect was suppressed after exercise for TAS and uric acid. In conclusion, the present study confirms diurnal variation of Hcy, selected biological markers of cellular damage, and antioxidant status in young football players. Also, the higher performances and muscle fatigue showed in the evening during RSA exercise might be due to higher levels of biological markers of muscle injury and lower antioxidant status at this TOD.     

Chemical synthesis, molecular modeling, and antimicrobial activity of a novel bacteriocin, MMFII

A new antimicrobial peptide, referred to as MMFII, was purified to homogeneity from lactic acid bacteria Lactococcus lactis, which were isolated from Tunisian dairy product. The complete amino acid sequence of the peptide has been established by amino acid analysis, Edman sequencing, and mass spectrometry and verified by solid-phase chemical synthesis. MMFII is a single-chain 37-residue polypeptide containing a single intramolecular disulfide bond, i.e., TSYGNGVHCNKSKCWIDVSELETYKAGTVSNPKDILW. It shares ca. 35% sequence identity with Leucocin A, a class IIa bacteriocin. Modeling based on the 3-D of Leucocin A shows three beta strands located in the N-terminal region (Thr1-Tyr3, Val7-Asn10, Lys13-Ile16) and an alpha helical domain from Asp17 to Asn31. When plotted as an alpha-helical wheel, the central alpha-helix of MMFII does not exhibit an amphipathic helical structure. The synthetic MMFII (sMMFII), obtained by the solid-phase method, was shown to be indistinguishable from the natural peptide. sMMFII is active against Lactococcus cremoris and Listeria ivanovii bacteria, whereas no activity was detected for any of the synthetic N-terminal truncated MMFII analogs Cys9-Trp37, Trp15-Trp37, and Val18-Trp37.     

Glutamate uptake in Lactobacillus delbrueckii subsp. bulgaricus CNRZ 208 and its enhancement by a combination of Mn2+ and Mg2+

AIMS: To demonstrate the mechanism of glutamate uptake in the dairy strain Lactobacillus delbrueckii subsp. bulgaricus CNRZ 208, and to characterize key aspects of the system. METHODS AND RESULTS: Glutamate uptake proceeded via an active transport system requiring an exogenous source of energy. The system also transported aspartate and glutamine. It was unique, with a Kt of 2.8 micro mol l-1 and a Vmax of 900 micro mol s-1 (g dry weight)-1. The activity was optimal at pH 7.3 and 50 degrees C, was independent of the glutamate charge, and was enhanced by Mn2+ + Mg2+ in combination. Inhibition of the activity by uncouplers and ionophores showed that transport was driven by an ATP-dependent mechanism involving the proton-motive force. This inhibition was partially abolished in the presence of both Mn2+ and Mg2+. CONCLUSIONS: We demonstrated for the first time that an active transport system governs the uptake of the essential amino acid glutamate in Lact. delbrueckii subsp. bulgaricus CNRZ 208, the activity of which is enhanced by a combination of Mn2+ and Mg2+. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the growth of Lact. delbrueckii subsp. bulgaricus in mixed-strain cultures for the dairy industry.     

Valine transport and biodiversity of Leuconostoc wild strains from French raw milk cheeses

The rate of L-valine transport in whole cells of Leuconostoc was at the maximum at 30 degrees C, pH 6.0 in the presence of an energy source. Transport was inhibited by 40-55%, in the presence of the ionophores (valinomycin, nigericin or monensin), and uncouplers (carbonyl cyanide-m-chloro-phenylhydrazone or 2,4-dinitrophenol) confirming the previously described delta p-driven branched-chain amino acid transport system described in cytoplasmic membranes (Winters et al., 1991, Appl. Environ. Microbiol., 57, 3350-3354). Sulfhydryl group reagents (p-chloro-mercuribenzoate, iodoacetate and N-ethyl maleimide) all inhibited valine transport by 60-70%, indicating that valine is actively transported at high valine concentration. Three kinetically distinguishable transport systems were identified for each strain using whole cells, confirming results obtained with membranes. L-valine transport Kt and Vmax could be an additional tool to estimate the biodiversity of 18 Leuconostoc strains belonging to the dominant flora of French raw milk cheeses. Kt values varied from 20 to 510 nmol/l for the very high affinity system, from 26 to 427 pmol/l for the high affinity system and from 0.65 to 4.40 mmol/l for the low affinity system. No correlation existed between valine transport rates and a particular strain's ability to acidify milk or complex media, suggesting that valine transport is not a growth-limiting function in species of the genus Leuconostoc.     

Risk factors in autism spectrum disorder: A Tunisian case-control study

BackgroundAutism spectrum disorder (ASD) is a neurodevelopmental condition that causes disability in social interaction, communication, and restrictive and repetitive behaviors. Common environmental factors like prenatal, perinatal, and/or postnatal factors play a key role in ASD etiologies. Moreover, specific metabolic disorders can be associated with ASD.Subjects and methodsWe performed a retrospective case-control study in child psychiatry clinics, involving 51 children with ASD and 40 typical development controls (TDC).ResultsWe found a correlation between children being breastfed for less than 6 months, having fathers more than 40 years old at childbirth in ASD compared to TDC group. Our study also associated low blood cholesterol and low erythrocyte magnesium levels with increased risk for ASD.ConclusionFindings support the implication of total cholesterol (TC) and erythrocyte magnesium level in defining autism outcome.     

A one-step reaction for the rapid identification of Lactobacillus mindensis, Lactobacillus panis, Lactobacillus paralimentarius, Lactobacillus pontis and Lactobacillus frumenti using oligonucleotide primers designed from the 16S-23S rRNA intergenic sequences

Aims:  Species-specific primers targeting the 16S–23S ribosomal DNA (rDNA) intergenic spacer region (ISR) were designed to rapidly discriminate between Lactobacillus mindensis , Lactobacillus panis , Lactobacillus paralimentarius , Lactobacillus pontis and Lactobacillus frumenti species recently isolated from French sourdough.
Methods and Results:  The 16S–23S ISRs were amplified using primers 16S/p2 and 23S/p7, which anneal to positions 1388–1406 of the 16S rRNA gene and to positions 207–189 of the 23S rRNA gene respectively, Escherichia coli numbering (GenBank accession number V00331 ). Clone libraries of the resulting amplicons were constructed using a pCR2·1 TA cloning kit and sequenced. Species-specific primers were designed based on the sequences obtained and were used to amplify the 16S–23S ISR in the Lactobacillus species considered. For all of them, two PCR amplicons, designated as small ISR (S-ISR) and large ISR (L-ISR), were obtained. The L-ISR is composed of the corresponding S-ISR, interrupted by a sequence containing tRNA^Ile and tRNA^Ala genes. Based on these sequences, species-specific primers were designed and proved to identify accurately the species considered among 30 reference Lactobacillus species tested.
Conclusions:  Designed species-specific primers enable a rapid and accurate identification of L. mindensis , L. paralimentarius , L. panis , L. pontis and L. frumenti species among other lactobacilli.
Significance and Impact of the Study:  The proposed method provides a powerful and convenient means of rapidly identifying some sourdough lactobacilli, which could be of help in large starter culture surveys.     

Viability Analysis of Fisheries Management on Hermaphrodite Population

We study the viability domains of bio-economic constraints for fishing model of hermaphrodite population, displaying three stages, juvenile, female and male. The dynamic of this model is subject to two constraints: an ecological constraint ensuring the stock perennity, and an economic constraint ensuring a minimum revenue for fishermen. Using viability kernel, we find out a viability domain which simultaneously guarantees a minimum stock level and a minimum income for fleets.