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High population of Sumateran wild boar causes the increasing number of hunting and overflowing of wild boar meat availability in the market at a very low price. This condition leads to the falsification of meat. This study was aimed to characterize and differentiate Sumateran wild boar meat (Sus scrofa vittatus) and beef (Bos taurus) since Sumateran wild boar meat is often falsified as a beef in Indonesian traditional market. The domesticated pork meat (Sus scrofa domestica) was also investigated to compare Sumateran wild boar meat and domestic pork. Samples used in this study were DNA (deoxyribonucleic acid) from pork (Sus scrofa domestica), Sumateran wild boar meat (Sus scrofa vittatus), and beef (Bos taurus). Beef and pork were obtained from traditional markets in Bogor city whereas Sumateran wild boar meat was obtained from Way Kanan Regency, Lampung Province. Herein, we performed a thorough investigation on Sumateran wild boar meat (Sus scrofa vittatus) and beef (Bos taurus) using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. This study utilized cytochrome b primer from mitochondria DNA with the size target amplification of 359 bp. This study utilized the restriction endonuclease enzymes, i.e. AluI, BsaJI, HindIII, RsaI, HaeIII, and TaqαI, in order to digest the amplification products from cytochrome b gene. The results showed that HaeIII is the merely specific enzyme to discriminate wild boar meat, domestic pork, and beef since HaeIII was able to digest these three meats in different locations of cyt b gene. However, AluI, BsaJI, HindIII, RsaI, and TaqαI enzymes were not specific enough to differentiate wild boar meat and domestic pork. In conclusion, among six tested enzymes, the use of PCR-RFLP analysis of the cyt b gene followed by digestion using HaeIII restriction enzyme provides a simple, relatively quick, and accurate identification of Sumateran wild boar species.

Lipoprotein lipase (LPL) is a key enzyme in lipid and lipoprotein metabolism. The canonical role of LPL involves the hydrolysis of triglyceride-rich lipoproteins for the provision of FFAs to metabolic tissues. However, LPL may also contribute to lipoprotein uptake by acting as a molecular bridge between lipoproteins and cell surface receptors. Recent studies have shown that LPL is abundantly expressed in the brain and predominantly expressed in the macrophages and microglia of the human and murine brain. Moreover, recent findings suggest that LPL plays a direct role in microglial function, metabolism, and phagocytosis of extracellular factors such as amyloid- beta (Aβ). Although the precise function of LPL in the brain remains to be determined, several studies have implicated LPL variants in Alzheimer’s disease (AD) risk. For example, while mutations shown to have a deleterious effect on LPL function and expression (e.g., N291S, HindIII, and PvuII) have been associated with increased AD risk, a mutation associated with increased bridging function (S447X) may be protective against AD. Recent studies have also shown that genetic variants in endogenous LPL activators (ApoC-II) and inhibitors (ApoC-III) can increase and decrease AD risk, respectively, consistent with the notion that LPL may play a protective role in AD pathogenesis. Here, we review recent advances in our understanding of LPL structure and function, which largely point to a protective role of functional LPL in AD neuropathogenesis.

The 1691 (G>A) factor V Leiden (FVL) and 20210 (G>A) prothrombin (PT) mutations are the two most common genetic risk factors in venous thromboembolism. The 677 (C>T) methylene tetrahydrofolate reductase (MTHFR) mutation is the most frequently mentioned as an independent genetic risk factor for venous thromboembolism. As there are limited published data on the prevalence of the 1691, 20210 and 677 mutations in our population, the aim of this study was to determine the frequencies and association of these deep vein thrombosis mutations in the Bosnian population. This study included 111 thromboembolic patients and 207 healthy subjects with absence of known risk factors for venous thromboembolism. Genotyping of the 1691, 20210 and 677 mutations was done by polymerase chain reaction (PCR), followed by restriction digestion with MnlI, HindIII and HinfI enzymes. Out of the 111 patients, 18.0% were heterozygous and 2.70% were homozygous for the 1691 mutation. Among 207 healthy controls, 3.86%, were heterozygous for the 1691 mutation. This study confirmed the association of the 1691 mutation with deep vein thrombosis in the Bosnian population odds ratio (OR) [95% confidence interval (CI)] = 6.0 (2.62-14.14); p = 0.0001). The 20210 mutation was detected in 2.70% of patients and it was totally absent in the control group. Allele and genotype frequency of 677 did not differ significantly between the cases and controls (χ2 = 1.03; p = 0.309).

This study aimed to evaluate the resolution of PCR-RFLP and sequencing of trnS/trnfM in detecting genetic diversity of seagrass Enhalus acoroides at Sanur Beach, Sindhu Beach and Semawang Beach. This research used six samples E. acoroides from each location. DNA extraction method followed Doyle and Doyle. PCR amplifications were done using primers P672/P673, P674/P675 and ITS4/ITS5 followed by digestion using restriction enzymes EcoRI, PstI, HindIII, BamHI, Rsa, Mva and HinfI. Sequencing analysis used PCR products of primers trnS/trnfM. Testing of DNA sequences of E. acoroides were conducted using BLAST (Basic Local Alignment Search Tool). DNA sequences were further analyzed using MEGA 5.2 software (Molecular Evolutionary Genetic Analysis) to evaluate variations of DNA. The sequence alignments were done using ClustalW software to determine the homology between the DNA sequences. The results showed that 18 samples of E. acoroides from Sanur, Sindhu and Semawang Beach have no polymorphism based on restriction enzyme analyses. Furthermore, sequencing of trnS/trnfM region of 18 samples E. acoroides showed that the sequences were identical.

Aims We evaluated the contributions of four polymorphisms of the lipoprotein lipase (LPL) gene to the risk of Alzheimer’s disease (AD). Methods Through a comprehensive literature search for genetic variants of LPL involved in AD association studies, we found four polymorphisms for the current meta-analyses. These polymorphisms were Asn291Ser(rs268), PvuII(rs285), HindIII(rs320) and Ser447Ter(rs328). Results In total, eight studies with 5064 cases and 5016 controls were retrieved for the meta-analyses of the four genetic variants. The analyses showed that Asn291Ser(rs268) (OR = 2.34, 95% CI = 1.05–5.25, P = 0.04), HindIII(rs320) (OR = 1.44, 95% CI = 1.17–1.78, P = 0.0006), and Ser447Ter(rs328) (OR = 0.80, 95% CI = 0.66–0.98, P = 0.03) were significantly associated with a risk of AD. No association was found between the PvuII(rs285) polymorphism and the risk of AD. Conclusions Our results showed that Asn291Ser(rs268), HindIII(rs320) and Ser447Ter(rs328) polymorphisms of LPL were associated with a risk of AD. Asn291Ser(rs268) and HindIII(rs320) were predisposing factors of AD, whereas Ser447Ter(rs328) showed a protective effect for AD.

Aim : in women with Hashimoto's disease (HD) we studied the clinical and some molecular genetic characteristics of the lipid profile. Materials and Methods : Subjects for this study included 109 women with HD. The average age of 57.2 ± 7.8 years, disease duration 8.0 ± 6.4 years, menopause duration 6.4 ± 3.5 years. Control group consisted of 85 women of similar age without thyroid pathology. Clinical and anthropometric data was collected from all participants. Whole blood samples were drawn in the morning after an overnight fasting for the measurement of serum TSH, free thyroxine (FT4), anti-thyroid peroxidase antibody (TPO-Ab) levels, as well as lipid concentrations and glucose. In genetic research participated 441 women: 104 women with HD and 337 healthy women (control subjects). The genotype of the subjects for TaqIB polymorphism of CETP, rs320 (HindIII +/-) polymorphism of LPL, polymorphism of the coding portion of the gene APOE, rs2228314 (1784G/C) polymorphism of SREBF2 was analyzed by using polymerase chain reaction-restriction fragment length polymorphism. Results : Stratified by triglycerides (TG) level increased in subjects with HD and lowered levels of cholesterol of high density lipoproteins (HDL). Similar results were obtained when comparing the indexes of blood lipids in the group of women who have reached the level of TSH in the range of 0.4–2.5 mU/l on the background of ongoing substitution therapy with thyroid hormone. Not received the differences in the distribution of genotypes of the polymorphisms of the gene CETP Taq1B, rs320 LPL gene and ɛ2/ɛ3/ ɛ4 APOE gene, rs2228314 SREBF2 gene in the main and control groups. B1B1 TaqIB polymorphism of CETP of the main group had the higher levels of TG and BMI. ɛ3/ɛ4 APOE had the higher levels of total cholesterol, LDL cholesterol, cholesterol non-HDL in women with hypothyroidism. Conclusion : In women with compensated hypothyroidism TG levels above and below the level of HDL cholesterol compared to the control. Shown association of APOE and CETP gene with some lipids parameters.

Background: Studies showed that lipid metabolism disorders are significant risk factors for myocardial infarction and coronary artery disease (CAD). Therefore, genes involved in lipid and lipoprotein metabolism pathways such as lipoprotein lipase (LPL), are proper candidates for susceptibility to CAD.Aim: To investigate the possible association between LPL gene variants (HindIII (rs320) and PvuII (rs285)), acute myocardial infarction (AMI) and serum lipid levels.Subjects and methods: The study population consisted of 211 patients with a diagnosis of premature AMI, and 203 age-matched individuals with normal coronary angiograms as controls. Genotyping of HindIII and PvuII polymorphisms was done by the PCR-RFLP technique. Results: Although the H+ and P+ alleles were more observed among the patients, there were no significant differences in genotype distributions and allele frequencies of HindIII and PvuII polymorphisms between patient and control subjects (P > 0.05). Triglyceride levels were found to be significantly elevated in H+H+ and P+P+ genotypes compared to others (P

This study targeting the characterization of new lytic phages and studying their impact on Staphylococcus aureus biofilm and growth in milk. Two lytic phages (vB_SauM_ME18 & vB_SauM_ME126) were isolated using 40 Multidrug-Resistant (MDR) S. aureus isolates. Four isolates (10%) were susceptible to phages. Phages were related to Family Myoviridae, had icosahedral heads and long contractile tails. The genome size is 20 kbp for both phages and DNA was digested with hinf I, hind III and hae III. Phages had a latent period (~15 min), rise period (~30 min) and burst size (114 PFU/infected cell for ME18 and 140 PFU/infected cell for ME126). There was a significant reduction in biofilm formation and reduction in the established biofilm biomass in phage treated S. aureus compared with controls (P

The complement receptor 1 (CR1) gene was shown to be involved in Alzheimer&rsquo, s disease (AD). We previously showed that AD is associated with low density of the long CR1 isoform, CR1*2 (S). Here, we correlated phenotype data (CR1 density per erythrocyte (CR1/E), blood soluble CR1 (sCR1)) with genetic data (density/length polymorphisms) in AD patients and healthy controls. CR1/E was enumerated using flow cytometry, while sCR1 was quantified by ELISA. CR1 polymorphisms were assessed using restriction fragment length polymorphism (RFLP), pyrosequencing, and high-resolution melting PCR. In AD patients carrying the H allele (HindIII polymorphism) or the Q allele (Q981H polymorphism), CR1/E was significantly lower when compared with controls carrying the same alleles (p &lt, 0.01), contrary to sCR1, which was significantly higher (p &lt, 0.001). Using multivariate analysis, a reduction of 6.68 units in density was associated with an increase of 1% in methylation of CR1 (estimate &minus, 6.68, 95% confidence intervals (CIs) &minus, 12.37, &minus, 0.99, p = 0.02). Our data show that, in addition to inherited genetic factors, low density of CR1/E is also acquired. The involvement of CR1 in the pathogenesis of AD might be linked to insufficient clearance of amyloid deposits. These findings may open perspectives for new therapeutic strategies in AD.

This study aimed to investigate the presence and diversity of AmpC β-lactamase and integrase genes among DNA (genomic and plasmid) from bacterial populations in selected aquatic systems. Following an enrichment step, DNA was isolated and subjected to polymerase chain reaction (PCR) and digital droplet PCR. The intI1 gene and AmpC β-lactamase genes were present in genomic and plasmid DNA from all sites in the Mooi, Crocodile and Marico Rivers, with the exception of intI1 in the Marico River. Digital droplet PCR demonstrated that copy numbers varied considerably (0.0 to 29.38 copies per picogram of DNA). Some samples in which ampC was not detected, intI1 was present. Amplicons of ampC genes were subjected to restriction digest using HindIII. Samples where the restriction markers were absent were purified by cloning followed by plasmid extraction, PCR amplification, and sequencing of individual AmpC gene fragments. Phylogenetic analysis identified all positive AmpC genes as Class C β-lactamases, comprising of ampC, CMY- and ACT-families. Detecting AmpC and intl1 genes on plasmids suggests a high risk of horizontal gene transfer and potential dissemination of these and other antibiotic resistance genes surrounding immediate aquatic environments. Consequences of β-lactamase diversity in aquatic ecosystems are relatively unexplored in South African aquatic ecosystems.

Lipoprotein lipase (LPL) is widely linked to lipid and lipoprotein metabolism, but its effects on coronary artery disease (CAD) are not clearly elucidated. The aim of the present study was to clarify the association between LPL gene polymorphisms and CAD susceptibility. The pooled odds ratio (OR) and 95% confidence interval (CI) were calculated to estimate the strength of the relationship between LPL gene polymorphisms and CAD risk. Comprehensive electronic databases, including PubMed, EMBASE, Web of Science, and the Cochrane Library, were systematically searched. A total of 45 records containing 80 eligible studies were analyzed. The results indicated an increased risk between the LPL D9N polymorphism and susceptibility to CAD in the dominant genetic model (AA + GA vs. GG: OR = 1.46, 95% CI = 1.14–1.87), whereas the LPL HindIII polymorphism showed a protective effect against CAD under all tested models (GG + GT vs. TT: OR = 0.85, 95% CI = 0.75–0.97; GG vs. TT + TG: OR = 0.62, 95% CI = 0.47–0.83; G vs. T: OR = 0.81, 95% CI = 0.71–0.92). No significant association was identified for the LPL N291S and PvuII polymorphisms. Stratification analysis by ethnicity suggested a significant correlation between the LPL S447X polymorphism and CAD susceptibility in Caucasians under the dominant and allele genetic models. In summary, our meta-analysis indicated that the LPL D9N polymorphism was associated with an increased risk of CAD, whereas the S447X and HindIII polymorphisms showed protective effects. There was no association observed between the N291S and PvuII polymorphisms and CAD risk.

onalan, ebru/0000-0001-9968-8201 WOS: 000428836700002 The objectives of this study were to detect kappa-casein (kappa-CN, CSN3, CASK) gene polymorphisms and to investigate the association between the kappa-CNgene polymorphisms and milk yield and composition in cattle. For this purpose, a total of 365 blood and milk samples were collected from Holstein, Jersey and Brown Swiss cows. Polymorphisms of kappa-CN gene were detected by the PCR and restriction fragment length polymorphism. A776-bp fragment of exon IV was amplified by PCR and digested with HindIII and HaeIII restriction endonuclease to distinguish allele A, B and E. The analysis of associations between kappa-CN genotypes and milk yield and composition were conducted with the use of the general linear model procedure. As a result, allele frequencies were detected as 0.736, 0.357, 0.308 for A allele, 0.186, 0.643, 0.692 for B allele in Holstein, Brown Swiss and Jersey breeds, respectively, and 0.079 for E allele in Holstein. This study also demonstrated the presence of Eallele in Turkish Holstein. Although kappa-CN genotypes were associated with protein and solids-not-fat content of milk in Hostein and Brown Swiss, they were associated with milk fat content in Jerseys (P < 0.05). Scientific and Technological Research Council of TurkeyTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TOVAG-107O612] The authors gratefully acknowledge the financial support from The Scientific and Technological Research Council of Turkey (Project No: TOVAG-107O612).

Introduction: Several polymorphisms at the lipoprotein lipase (LPL) locus are associated with variations in LPL activity serum lipid concentrations and the risk of coronary artery disease (CAD). The aim of this study was to investigate the role of the LPL S447X and Hind III polymorphism in a sample of subjects with CAD and compare them with healthy subjects. Methods: The study enrolled 115 patients and 89 healthy subjects who were recruited from Namazi hospital in 2010-2012. The presence of two common polymorphisms of the LPL gene(HindIII and S447X) was determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis using genomic DNA. SPSS 16.0 was used for statistic alanalysis. Results: S447X was significantly different between the patients with CAD and the healthy subjects (P

Aims: Growing incidences of the pathogens producing Metallo-β-lactamases (MBLs) has been observed in many countries including India. They are carbapenemases that are capable of hydrolysing carbapenems that are often considered as a last resort antibiotic, for infections caused by multiple drug resistant bacteria. The objective of the current study was to investigate the prevalence of New Delhi Metallo-β-lactamase (NDM-1) and other MBL producers among 747 uropathogens collected from Mumbai city. Place and Duration of Study: The current study was carried out at Department of Microbiology, Wilson College, Mumbai 400007 between September 2011January 2014. Methodology: The screening of MBL producers was done using phenotypic detection tests such as antimicrobial susceptibility test, Modified Hodge Test and MBL E-Test. Molecular characterization of the test isolates was carried out using Molecular tests like Polymerase chain reaction and Restriction enzyme digestion. Results: In the current study, 49 MBL producers were obtained, all of which harboured a plasmid of Original Research Article Mobashshera and Aruna; BMRJ, 5(4): 368-383, 2015; Article no.BMRJ.2015.038 369 approximately 50 kb size. Polymerase chain reaction showed the presence of blaNDM-1 in 43/49, blaVIM-1 in 3/49, and blaIMP gene in 3/49 MBL producers. Co-production of Extended Spectrum βLactamase namely, blaTEM and blaCTX-M genes were also observed among 25/49 MBL producers. Determination of Minimum Inhibition Concentration (MIC) showed only 35/49 MBL producers to be resistant to meropenem, indicating difficulty in routine MBL detection where it is generally carried out as a screening step. Plasmid borne resistance of MBL producers was confirmed by plasmid loss studies. Restriction enzyme analysis carried out with the help of Eco RI, Bam HI and HindIII revealed different band patterns among MBL producers, indicating clonal unrelatedness and more than one source of origin for spread of antibiotic resistance. Conclusion: The current study may serve as a basis for better understanding of the resistance patterns of MBL producers, and thus restrict the spread of drug resistance.

Hemophilia A (HA) is an inherited X-linked bleeding disorder caused by mutations in the factor VIII gene. Prenatal detection in female carriers from families with HA is important to reduce the number of HA patients. The purpose of this study was to detect carriers in families with HA from Sichuan, China, using linkage analysis and a direct genotyping method. A total of 18 HA families were studied. Using a combination of intron 22 inversion, intron 1 inversion, the BclI polymorphic site in intron 18, the HindIII polymorphic site in intron 19, and dinucleotide CA-repeat markers in introns 1, 13, 22, and 24, we were able to detect HA in 88.9% (16/18) of the families studied. HA was detected in the remaining two families by direct genotyping. This study gave the participants a good understanding of their genetic condition and gave us a preliminary understanding of the prevalence of each mutation in Sichuan HA patients.

A time-resolved study using the freeze-trap method elucidates the mechanism of the DNA-cleaving reaction of HindIII., In order to investigate the mechanism of the reaction catalyzed by HindIII, structures of HindIII–DNA complexes with varying durations of soaking time in cryoprotectant buffer containing manganese ions were determined by the freeze-trap method. In the crystal structures of the complexes obtained after soaking for a longer duration, two manganese ions, indicated by relatively higher electron density, are clearly observed at the two metal ion-binding sites in the active site of HindIII. The increase in the electron density of the two metal-ion peaks followed distinct pathways with increasing soaking times, suggesting variation in the binding rate constant for the two metal sites. DNA cleavage is observed when the second manganese ion appears, suggesting that HindIII uses the two-metal-ion mechanism, or alternatively that its reactivity is enhanced by the binding of the second metal ion. In addition, conformational change in a loop near the active site accompanies the catalytic reaction.

Background: Single nucleotide polymorphism (SNP) 148C/T which is located in β fibrinogen gene (FGB) promoter has correlation with fibrinogen levels; however, the association of SNP 148C/T and ischemic stroke in young adult patients is contradictory. Aim: To determine the association of SNP 148C/T in FGB promoter with plasma fibrinogen levels and ischemic stroke in young adults. Subjects and methods: In this case-control study, SNP 148C/T among 107 ischemic stroke patients and 94 controls were evaluated by PCR-RFLP with restriction enzyme HindIII and confirmed by DNA sequencing. Physical and neurological examinations, brain computed tomography, plasma fibrinogen levels and blood biochemistry tests were assessed within seven days after the onset of symptoms. Genotype distributions and allele frequencies were analyzed by chi-squared test. Results : This study found that the level of fibrinogen was significantly higher in ischemic stroke group than control (419.2 mg/dL vs. 351.1 mg/dL, p < 0.000) and the level of fibrinogen associated with ischemic stroke (OR, 2.28; 95%CI, 1.28–4.07, p= 0.005). Mutant genotypes (CT and TT) and T allele had a significant association with hyperfibrinogenemia (OR, 2.58; 95%CI, 1.39–4.76 and OR, 1.6; 95%CI, 1.60–2.41, respectively) and ischemic stroke (OR, 2.46; 95%CI, 1.37–4.41 and OR, 1.80; 95%CI 1.19–2.73, respectively). In addition, analysis adjusted for other risk factors found that mutant genotypes correlated with hyperfibrinogenemia and ischemic stroke (OR, 2.27; 95%CI, 1.21–4.25 and OR, 2.16; 95%CI, 1.19–3.94, respectively). Conclusion : There was a significant association between SNP 148C/T and fibrinogen levels, SNP 148C/T and ischemic stroke, and fibrinogen levels and ischemic stroke. Keywords: Ischemic stroke; Fibrinogen; Hyperfibrinogenemia; 148C/T FGB polymorphism; rs1800787.

application/pdf, Monoclonal integration of bovine leukemia virus (BLV) proviral DNA into bovine genomes was detected in peripheral blood from two clinical cases of enzootic bovine leukosis (EBL) without enlargement of superficial lymph nodes. A BLV-specific probe hybridized with 1 to 3 EcoRI and HindIII fragments in these 2 atypical EBL cattle by Southern blotting and hybridization, as well as in 3 typical EBL cattle. The probe also hybridized to a large number of EcoRI and HindIII fragments in 5 cattle with persistent leukosis. These results suggest that the detection of monoclonal integration of BLV provirus into the host genome may serve as a marker of monoclonal proliferation and malignancy in difficult to diagnose EBL cattle.

Bacteriophage named as SAL-PG ( Salmonella bacteriophage ), specific to Salmonella pullorum and Salmonella gallinarum was isolated by an enrichment method directly from waste water near to poultry farm. For the isolation of phage 4.5 ml water sample, 0.5 ml of 10X Nutrient Broth and 0.5 ml of log phage bacteria were mixed well and incubated at 37° C for overnight followed by centrifugation at 10,000 rpm for 10 minutes. The presence of bacteriophage was observed by spot test over the bacterial lawn of and plaque assay. The host range of the isolated bacteriophage was determined by spot test using 12 different bacterial isolates. The phages were found to infect Salmonella pullorum , Salmonella gallinarum and Salmonella typhimurium and produced clear plaque on these Salmonella serovars. The bacteriophage was able to survive in wide range of pH between 2 to 9 and resistant at 60° C for 1 hour. More than 50% phages were readily adsorbed to the host bacteria. The restriction enzyme analysis showed that the phage DNA possess restriction site for TaqαI, HindIII and BstYI; but no restriction site for XhoI and BstEII as these two enzymes failed to digest the phage DNA. The characterizations of the bacteriophage would be helpful in establishing a basis for adopting the application of the most effective bacteriophage treatment to control these poultry bacteria. DOI: http://dx.doi.org/10.3329/bjvm.v12i2.21264 Bangl. J. Vet. Med . (2014). 12 (2): 107-114

We constructed a new Thermus thermophilus cloning vector which enables the colour selection of cloned DNA inserts in the T. thermophilus HB27 host strain (β-gal-) on growth plates containing 3,4-cyclohexenoesculetin β-D-galactopyranoside (S-gal) in the medium. This vector harbors a modified β-galactosidase gene (TTP0042 of T. thermophilus HB27) with 12 unique restriction enzyme sites (Acc65I, AvrII, BlpI, BssHII, EcoRI, EcoRV, HindIII, NruI, SalI, SpeI, SphI and XbaI) as multiple cloning sites under the control of the T. thermophilus slpA promoter. This host-vector system facilitates cloning procedures in T. thermophilus HB27.

Sickle cell disease shows several clinical manifestations in distinct levels of severity. This heterogeneity is due to the haplotype variability associated with the HbS gene, levels of fetal hemoglobin and environmental conditions, which modify the disease expression. Science community believes that the presence of a polymorphism in the CCR5 gene, which is related to chronic inflammatory state, could confer a higher survival rate and a lower number of inflammatory events to these patients since the deletion in CCR5Δ32 would knock out the CCR5 gene. Therefore, this study aimed to identify the haplotypes in βS and βC genes, as well as to investigate the presence of the CCR5Δ32 deletion in patients with sickle cell disease. For this purpose, DNA was isolated with the QIAamp DNA Investigator Kit, and PCR was the method chosen to detect the mutant allele CCR5Δ32. The haplotypes in βS and βC genes were detected by RFLP with the restriction enzymes XmnI, HindIII, HincII, and HinfI analyzing six polymorphic sites on the β cluster, succeeded by electrophoresis. The atypical haplotype was the most common (54.3%), followed by Benin (28.6%), Bantu (11.5%), Senegal (2.8%), and Cameroon (2.8%). No patients presented CCR5Δ32 deletion. The increase in the frequency of atypical haplotypes suggests that these patients passed by variation in the genetic pattern from ancestral haplotypes throughout the years.

PubMedID: 28881255 The aim of this study was to synthesize and evaluate plasmid DNA interaction of new platinum(II) complexes with some 2-substituted benzimidazole derivatives as carrier ligands which may have potent anticancer activity and low toxicity. Twelve benzimidazole derivatives carrying indole, 2-/or 3-/or 4-methoxyphenyl, 4-methylphenyl, 3,4-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 4-methoxybenzyl, 3,4,5-trimethoxybenzyl, 3,4,5-trimethoxystyryl, 3,4,5-trimethoxybenzylthio or dimethylamino ethyl groups in their position 2 and twelve platinum(II) complexes with these carrier ligands were synthesized. The chemical structure of the platinum complexes have been characterized by their elemental analysis and FIR, 1H NMR and mass spectra and their 1H NMR and FIR spectra were interpreted by comparison with those of the ligands. The interaction of all the ligands and their complexes with plasmid DNA and their restriction endonuclease reactions by BamHI and HindIII enzymes were studied by agarose gel electrophoresis. It was determined that complex 1 [dichloro-di(2-(1H-indole-3-yl)benzimidazole)platinum(II)·2H2O] has stronger interaction than carboplatin and complex 10 [dichloro-di(2-(3,4,5-trimethoxystyryl)benzimidazole)platinum(II)·2H2O] has stronger interaction than both carboplatin and cisplatin with plasmid DNA. © 2017 Elsevier Inc. BAP-02/2015-02 We would like to thank the Research Foundation of Gazi University ( BAP-02/2015-02 ) for financial support of this work.

Ecological damages to the Lagoon of Segara Anakan, Central Java, as well as large-scale and continuous exploitation are threatening the sustainability of fine shrimp, Metapenaeus elegans, and resources. Information in regards to genetic resources is crucial to establish long-term conservation programs and to preserve germplasm quality. This study aims to evaluate the number and size of the fragment which is digested with restriction enzyme Hind III. Seven individuals of Metapenaeus elegans from the Lagoon of Segara Anakan were examined using Hind III. Amplification of mitochondrial DNA resulted in 950 bp, and the digestion using Hind III generated four fragments consisting of 114 bp, 200 bp, 250 bp, and 386 bp, which formed a monomorphic pattern. The restriction pattern showed the probability of homozygosity of alleles that restricted using Hind III. Homozygosity indicates no variation of DNA sequence.

Background: In recent years, the lipoprotein lipase (LPL) polymorphism has been extensively investigated as a potential risk factor for coronary artery disease (CAD). However, the results of these studies have been inconsistent. Therefore, we performed this meta-analysis to explore the association between LPL polymorphism and CAD risk. Methods: The literature was searched from electronic databases such as Embase, China Biological Medicine Database, PubMed, Knowledge Infrastructure, and China National Web of Science by the key words “coronary artery disease”, “lipoprotein lipase” and “polymorphism”. All of the studies included in this manuscript met the inclusion and exclusion criteria. An odds ratio (OR) analysis using a 95% confidence interval (CI) was employed to assess the association of the LPL polymorphism with CAD susceptibility. Results: We performed a meta-analysis of 14 case-control studies including HindIII, Ser447X and PvuII polymorphism. A statistically significant increase in the risk of CAD was associated with LPL HindIII polymorphism. This included HindIII H+H+ genotype (OR = 1.28, 95% CI = 1.09–1.49, p = 0.002, I2 = 43%) and H+ allele genotype (OR = 1.27, 95% CI = 1.03–1.58, p = 0.03, I2 = 67%). Ser447X XX genotype (OR = 2.37, 95% CI = 1.33–4.24, p = 0.004, I2 = 53%) was also associated with CAD risk. However, PvuII polymorphism was found to have no significant association with CAD risk. Conclusions: LPL HindIII polymorphism was significantly associated with the risk of CAD. For Ser447X polymorphism, it was found that only XX genotype was significantly associated with CAD risk. Furthermore, PvuII polymorphism had no significant association with CAD risk. It was considered that LPL HindIII polymorphism might serve as a potential biomarker for CAD risk.

Hemophilia A (HA) is an inherited X-linked coagulation disorder caused by the deficiency of factor VIII ( FVIII). Linkage analysis is a common indirect method for the detection of female carriers in families with HA. In the current study, 173 patients from 30 unrelated families with HA were recruited from the Azeri Turkish population of northwest Iran and analyzed for BclI and HindIII markers by polymerase chain reaction-restriction fragment length polymorphism. We investigated the potential of using these markers for the detection of mutation in carriers through linkage analysis, which would be of tremendous use in prenatal diagnosis. Among the tested women, 47% and 35% were found to be heterozygous for BclI and HindIII polymorphic markers, respectively. The BclI and HindIII markers were informative for the detection of 63% and 17% potential carriers, respectively, demonstrating the effectiveness of the BclI marker for the detection of HA carriers among the Azeri Turkish population.

Phenotypic identification of non-pylori Helicobacter species has always been problematic and time-consuming in comparison with many other bacteria. We developed a rapid two-step identification assay based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the 23S rRNA gene for differentiating between non-pylori Helicobacter species. A new genus-specific primer pair based on all available complete and partial 23S rRNA sequences of Helicobacter species was designed. In silico restriction analysis of variable regions of the 23S rRNA gene suggested SmaI and HindIII endonucleases would provide a good level of differentiation. Analysis of the obtained 23S rRNA RFLP patterns divided all Helicobacter study strains into three species groups (groups A-C) and 12 unique restriction patterns. Wolinella succinogenes also gave a unique pattern. Our proposed PCR-RFLP method was found to be as a valuable tool for routine identification of non-pylori Helicobacter species from human or animal samples.

El polimorfismo Hind III es la variante más común en el gen de la lipoproteinlipasa; sin embargo, su asociación con enfermedad cardiovascular es controversial. Objetivo: Establecer la frecuencia del polimorfismo Hind III y su relación con el perfil lipídico y la enfermedad obstructiva coronaria (EOC) en pacientes colombianos. Materiales y métodos: La muestra la constituyeron pacientes que asistieron a un centro de hemodinamia del Quindío, por necesidad de una angiografía coronaria. El polimorfismo Hind III fue evaluado por la reacción en cadena de la polimerasa y restricción enzimática. Resultados: 389 pacientes fueron divididos en individuos con EOC≥50 %, (60,4 %) e individuos con EOC

Made available in DSpace on 2015-03-18T15:55:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-01Bitstream added on 2015-03-18T16:28:39Z : No. of bitstreams: 1 WOS000342939400001.epub: 823912 bytes, checksum: 01aa50918dc647fba6aa7a39fd6bb571 (MD5)Bitstream added on 2015-03-18T16:28:39Z : No. of bitstreams: 2 WOS000342939400001.epub: 823912 bytes, checksum: 01aa50918dc647fba6aa7a39fd6bb571 (MD5) WOS000342939400001.pdf: 1333648 bytes, checksum: f0c8479a8a4f4251a7f53dbc533dd94e (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Laboratorio de Investigacao Medica In visceral leishmaniasis (VL) endemic areas, a minority of infected individuals progress to disease since most of them develop protective immunity. Therefore, we investigated the risk markers of VL within nonimmune sector. Analyzing infected symptomatic and, asymptomatic, and noninfected individuals, VL patients presented with reduced high-density lipoprotein cholesterol (HDL-C), elevated triacylglycerol (TAG), and elevated very-low-density lipoprotein cholesterol (VLDL-C) levels. A polymorphism analysis of the lipoprotein lipase (LPL) gene using HindIII restriction digestion (N = 156 samples) (H+ = the presence and H = the absence of mutation) revealed an increased adjusted odds ratio (OR) of VL versus noninfected individuals when the H+/H+ was compared with the H-/H-genotype (OR = 21.3; 95% CI = 2.32-3335.3; P = 0.003). The H+/H+ genotype and the H+ allele were associated with elevated VLDL-C and TAG levels (P < 0.05) and reduced HDL-C levels (P < 0.05). An analysis of the L162V polymorphism in the peroxisome proliferator-activated receptor alpha (PPAR alpha) gene (n = 248) revealed an increased adjusted OR when the Leu/Val was compared with the Leu/Leu genotype (OR = 8.77; 95% CI = 1.41-78.70; P = 0.014). High TAG (P = 0.021) and VLDL-C (P = 0.023) levels were associated with susceptibility to VL, whereas low HDL (P = 0.006) levels with resistance to infection. The mutated LPL and the PPAR alpha Leu/Val genotypes may be considered risk markers for the development of VL. Univ Sao Paulo, Inst Med Trop Sao Paulo, Lab Soroepidemiol & Imunobiol, BR-05403000 Sao Paulo, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Parasitol, BR-18618970 Botucatu, SP, Brazil Univ Fed Piaui, Inst Doencas Trop Natan Portella, BR-64001450 Teresina, PI, Brazil Univ Estado Rio de Janeiro, Inst Social Med, Dept Epidemiol, BR-20550900 Rio De Janeiro, RJ, Brazil Univ Fed Rio de Janeiro, Inst Estudos Saude Colet, BR-21941598 Rio De Janeiro, RJ, Brazil Univ Sao Paulo, Fac Med, Dept Prevent Med, BR-01246903 Rio De Janeiro, RJ, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Parasitol, BR-18618970 Botucatu, SP, Brazil CNPq: 154581/2006-2 FAPESP: 06/60006-9 Laboratorio de Investigacao MedicaLIM/38 HC-FMUSP

Carrier frequency of the βS allele has been reported to be 0.19% in Mazandaran province, northern Iran. Haplotype analysis of the βS allele helps trace the origin of its encoded hemoglobin (Hb) variant, Hb S, in a region. The aim of this study was to investigate the haplotypes associated with βS alleles in Mazandaran province. Capillary electrophoresis was carried out to detect individuals suspected to have a βS allele(s). DNA analysis (PCR-RFLP) was used for final confirmation. To identify 5' to 3' β-globin gene cluster haplotypes associated with βS alleles, family linkage analysis was applied. Six polymorphic sites (HincII 5' to e, XmnI 5' to Gγ, HindIII in Gγ, HindIII in Aγ, HincII 3' to ψβ and AvaII in β) were investigated using the PCR-RFLP method. Five different haplotypes were linked to βS alleles, while βA alleles were associated with nine haplotypes. Among the βS alleles, 53.9% were associated with the Benin (----++) haplotype, and the Arab-Indian (+++-++) haplotype had the second-highest frequency (23%). Unlike southern provinces, where the Arab-Indian haplotype is prominent, the Benin haplotype is the most frequent haplotype in northern Iran, and this may represent a founder effect. Since the Benin haplotype does not carry the XmnI polymorphism 5' to the Gγ gene, which is responsible for high expression of Hb F, a severe form of sickle cell disease can be anticipated in patients that are homozygous for the βS allele in the northern region.

Background Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by immune complexes formation and body organs damage. Recently, plasminogen activator inhibitor-1 (PAI-1) has been proposed recently to modulate the immune response. Aim of the study To elucidate whether there is possible role of two genetic polymorphisms in the PAI-1 gene: −675 4G/5G and HindIII C>G in lupus nephritis (LN) disease in Egyptian patients. Patients and methods Using PCR-restriction fragment length polymorphism, we genotyped 170 Egyptian patients with SLE; 80 fulfill the American College of Rheumatology criteria of LN and 90 SLE patients without LN for two polymorphisms of PAI-1 gene: −675 4G/5G and HindIII C>G. Results The 4G4G genotype, 4G allele of PAI 4G/5G polymorphism, and the G allele of the PAI-1 HindIII C>G polymorphism, all exhibited significant risks for proliferative LN [odds ratio (OR)=2.55; 95% confidence interval (CI): 1.02–21.11; P=0.022); (OR=2.43; 95% CI: 1.18–5.01; P=0.015); (OR=2.37; 95% CI: 1.01–5.58; P=0.045), respectively]. Moreover, PAI-1 4G/5G polymorphism was related to the chronicity index for LN (P G polymorphism was associated with the activity (P=0.016) and the chronicity (P Conclusion Egyptian patients with SLE with a polymorphism for −675 4G/5G and HindIII C>G of PAI-1 gene might have a higher susceptibility for the development of proliferative LN.

Hemophilia A (HA) is the most common congenital X-linked coagulopathy caused by mutations in the factor VIII gene. One in 5000 to 10 000 male persons worldwide suffer from HA. It is the archetype of high-cost, low-volume disease. Therefore, identification of carriers is crucial to avoid the birth of affected males. Tracking of the defective X chromosome through indirect linkage analysis represents the most practical method for screening for carriers in developing countries. In this study, 227 individuals from 41 families with HA and 100 normal participants were recruited from the Kurdistan region of Iraq and evaluated for intron 18 BclI, intron 19 HindIII, and IVS7 nt 27 markers by polymerase chain reaction restriction fragment length polymorphism and direct sequencing. Among the studied women, 49%, 42%, and 14% were discovered to be heterozygous for BclI, HindIII, and IVS7 markers, respectively. Using BclI, HindIII, and IVS7 markers, 56%, 46%, and 17% of the families were informative, respectively. The combined informativity of these polymorphic sites reaches 66%. The current study illustrates the effectiveness of the BclI and HindIII markers for the diagnosis of HA carriers among the Iraqi Kurdish population.

The study describes a rapid approach for detection of common enteric bacterial pathogens, which involves partial amplification of the 16S rRNA gene by PCR using a colony from selective medium followed by restriction enzyme (RE) digestion using the EcoRI, HindIII and SalI enzymes. On the basis of RE digestion analysis different genera namely, Escherichia, Salmonella, Shigella, Vibrio, Campylobacter, Arcobacter, Yesinia and Listeria were differentiated.

Genetic polymorphism k-casein (CSN3) gene was investigated in lactating Egyptian buf- falo using nucleotide sequencing. Primer pairs amplified a 453 nucleotide fragment of CSN3 exon IV with an open reading frame of 421 nucleotides encoding 139 amino acids of the mature peptide and 32 nucleotides 3 0 UTR. Two SNPs (nt-315 C/T and nt-319 C/T) occurred in amplified fragment. These SNPs were reflected at codon 105 (ACC/ATC) and codon 106 (ACC/ACT) which correspond to codon 135 and 136 of the CSN3 mature peptide, respectively. Variation at codon 135 caused a change from ACC (Threonine) versus ATC (Isoleucine) whereas variation at codon 136(Thr/Thr) is a silent mutation. The results show, contrary to previous reports, that Egyptian buffalo has both alleles A (135 Thr ACC/136 Thr ACC) and B (135 Ile ATC/136 Thr ACT) with allelic frequencies of 0.57 and 0.43, respectively. The Egyptian buffalo genotype frequencies were 0.294 (AA), 0.647 (AB) and 0.058 (BB). The polymorphic site at codon 135 (A(C/T)C) and 136 (AC(C/T)) has C bases in allele A and T bases in allele B, resulting in two haplotypes; 135 Thr(ACC) /136 Thr(ACC) and 135 Ile(ATC) /136 Thr(ACT) . The frequency of the former haplotype was 0.57. In this study we investi- gated the reason why buffalo samples, analyzed by RFLP technique, using HindIII and HinfI used in cattle, were mistakenly identified as BB monomorphic. We suggest the use of restriction enzymes AcuI or Eco57MI to be used in buffalo CSN3 RFLP analysis. Digestion of buffalo CSN3-exon VI fragment (453 bp) with either enzyme will generate two fragments of 339 bp and 114 bp in allele B. a 2013 Production and hosting by Elsevier B.V. on behalf of Academy of Scientific Research & Technology.

Introduction: Studies have reported an association between lipoprotein lipase (LPL) gene and myocardial infarction in some populations. Therefore, the present study aimed to investigate the association of the HindIII polymorphism of the (LPL) gene with myocardial infarction and to explore its potential role in susceptibility in a South Indian population. Subjects and methods: We included a total of 412 subjects (202 myocardial infarction patients and 210 age- and sex-matched controls). Demographic and clinical characteristics were collected. Lipid profiles were estimated. DNA was isolated and the LPL gene HindIII polymorphism was determined by polymerase chain reaction. Results: Comparison of the lipid profiles between patients and controls showed that patients had statistically high significant values (p = 0.0001). The H+ H+ genotype of the LPL gene is associated with myocardial infarction. H+ H+ vs. H− H− was χ2 = 19.4, OR 3.1, CI 95% 1.8–5.2, p

Background: Coronary artery disease (CAD) is one of the leading causes of death and disability around the world. Interaction between genetic and environmental factors determines susceptibility of an individual to develop coronary artery disease . Lipoprotein lipase (LPL) play an important role in the metabolism of HDL-C ( High Density Lipoprotein Cholesterol ), LDL-C (Low Density Lipoprotein Cholesterol ) and triglycerides (TG). Dysfunction of LPL as a result of genetic variants of lipoprotein lipase gene is associated with increased risk of CAD. The aim of the present study was to investigate the relationship between the risk of coronary artery disease and LDL-C, HDL-C and TG (triglycerides) levels by lipoprotein lipase gene Hind III polymorphism. Materials and Methods: A total of 202 subjects including 114 patients with coronary artery disease and 88 control participated in this study. The Hind III polymorphism of the lipoprotein lipase gene was determined by PCR- RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) . In the presence and absence of restriction site, the genotypes are described H+/+ , H-/- respectively. Results: In this survey, a highly significant association between the frequent H+/+ genotype and unfavorable TG levels was observed in our population . For the Hind III genotypes, within the healthy subjects (n=88), the H+/+ genotype was found in 67 individuals (58.8%), H-/+ genotype in 38 individuals (33.3%) , and 9 individuals (7.8%) carried the H-/- genotype. Within the CAD group (n=114), 47 individuals (53.4%) with H+/+ genotype, 36 (41%) with H-/+ genotype, and 5 (5.6%) carried the H-/- genotype. Conclusion: There was a significant difference between the distribution of LPL–Hind III genotypes and the healthy subjects and the patients with CAD (P

Ecofriendly synthesis of metal nanoparticles and their unique properties hold promise for innovative therapeutic applications. In the present study, rapid, novel, low cost method for synthesis of silver nanoparticles (AgNPs) using aqueous leaves extract of medicinal plant Euphorbia heterophylla ( E. heterophylla ) is reported. AgNPs synthesized by plant extract exhibited absorption peak at 422 nm in UV–vis spectroscopy. Transmission electron microscopy micrographs revealed presence of AgNPs with average size of 13 nm with grain and triangular shapes. Fourier Transform Infrared Spectroscopy analysis revealed role of leaves metabolites as reducing and capping agent in synthesis of AgNPs. X ray diffraction pattern and selected area electron diffraction study confirmed crystalline nature of AgNPs. Inhibition of DNA cutting activity of restriction endonucleases ( EcoRI, HindIII and BamHI ) by biofunctionalized AgNPs in agarose gel electrophoresis is first time reported in the present study. This potential of AgNPs can be useful for enhancing effectiveness of phage therapy by acting as synergistic cocktail with bacteriophages to kill bacteria via inhibition of their restriction endonucleases.

Background It is known that patients with sickle cell disease (SCD) present activation of the blood coagulation and fibrinolytic systems, especially during vaso-occlusive crises and also during the steady state of the disease. We determined whether the presence of the factor prothrombin gene G20210A variant, factor V gene G1691A mutation (factor V Leiden), and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms may be risk factors for vascular complications in individuals with SCD. Methods The study involved 150 patients with sickle cell anemia and 150 healthy controls of Central India. Genotyping of three thrombophilic mutations was carried out by PCR-RFLP methods using MnlI, Hind III, and Hinf I, respectively, for factor V Leiden, prothrombin, and MTHFR mutations. Results Patients with SCD had significantly higher prevalence of mutant variants of MTHFR gene (28.0% heterozygotes and 14.6% homozygotes) and FVL gene (14.6% heterozygotes) as compared to normal/control individuals, but complete absence of mutant variants of prothrombin gene. The patients with SCD having mutant variants of MTHFR and FVL genes showed higher incidence of pain in chest, abdomen, and bone joints along with early age of onset of clinical manifestations as well as frequent dependence on blood transfusion than those patients with SCD having wild variants of these thrombotic genes. As compared to control subjects, SCD individuals having mutant variants of FVL and MTHFR genes had significant association with higher levels of prothrombin fragment (F1+2), D-dimer, thrombin-antithrombin (TAT), and lower level of protein C. Conclusion MTHFR C677T and FVL G1691A polymorphisms may be risk factors for increased vascular complications in patient with SCD.

Complement blockade by eculizumab is clinically effective in hemolytic paroxysmal nocturnal hemoglobinuria. However, the response is variable and some patients remain dependent on red blood cell transfusions. In 72 patients with hemolytic paroxysmal nocturnal hemoglobinuria on eculizumab we tested the hypothesis that response may depend on genetic polymorphisms of complement-related genes. We found no correlation between the complement component C3 genotypes and the need for blood transfusions. On the other hand, we found a significant correlation with the HindIII polymorphism of a complement regulatory gene, the complement receptor 1 (CR1) gene. At this locus two co-dominant alleles are known, of which H (common) is associated with high expression, whereas L (rare) is associated with low expression of CR1 on red blood cells. Patients who still needed blood transfusion on eculizumab accounted for 18% of the H/H homozygotes, 33% of the H/L heterozygotes and 68% of the L/L homozygotes (P=0.016). Thus, patients with paroxysmal nocturnal hemoglobinuria who have the L/L genotype are seven times more likely to be sub-optimal responders to eculizumab. Both in vitro and in vivo we found that the CR1 HindIII genotype correlates with the abundance of paroxysmal nocturnal hemoglobinuria red cells that have bound C3, and with the kinetics of C3 binding. These results are consistent with the notion that by affecting C3 binding the CR1 genotype influences the response to eculizumab treatment, and this emerges as a novel example of pharmacogenetics.

Based on the polymerase chain reaction (PCR), ag85a and mpb70 Fusion gene of Mycobacterium bovis were ligated and cloned into pMD18-T, then recombinant plasmid pMD-85a-70 was constructed. pMD-85a-70 and pVAX1-BMS were digested by double enzymes HindIII and EcoRI, the purified ag85a-mpb70 was subcloned into pVAX1-BMS, then recombinant plasmid pVAX1-BMS-85a-70 was constructed and transient expressed in Marc145 cell. These results laid solid foundations for further studies on ag85a-mpb70.

Objective The objective of this study was to investigate the effects of the interaction between lipoprotein lipase (LPL) Pvu II and Hind III haplotypes and carbohydrate intakes on the components of metabolic syndrome (MetSyn) in Koreans. Methods LPL Pvu II and Hind III genotype, LPL mass, triglyceride, high-density lipoprotein cholesterol, blood pressure (BP), waist circumference (WC), insulin, and Homeostasis Model of Assessment - Insulin Resistance were determined using a cross-sectional design in 269 controls and 280 MetSyn patients. Results LPL mass was significantly lower in patients with PvuII and HindIII mutant alleles (P2 and H2) and decreased as the number of MetSyn components increased in all Pvu II and Hind III haplotypes. Both LPL mass-adjusted WC and systolic BP (SBP) were positively associated with a ratio of percent energy from carbohydrate to percent energy from fat in individuals with P2H2 haplotype. After adjustment for age, sex, and LPL mass, the odds ratio (OR) for excessive WC was higher in carriers of P2H2 in the highest carbohydrate intake tertile compared with carriers of P1H1 in the lowest carbohydrate intake tertile (OR, 6.94; 95% confidence interval [CI], 1.39–34.62). Moreover, the OR for high SBP were higher in carriers of P1H2/P2H1 in the highest carbohydrate intake tertile (OR, 7.84; 95% CI, 1.79–34.46) and in carriers of P2H2 in the highest carbohydrate intake tertile (OR, 4.24; 95% CI, 1.16–15.48) than P1H1 carriers. Conclusion This study suggests that P2H2 carriers in the highest carbohydrate intake tertile may be at risk for MetSyn because they had increased odds of excessive WC and high SBP.

The gene of Mus musculus was cloned into plasmid vector (pYEUra3) of E.coli from the cDNA library which was then verified using colony PCR. The plasmid DNA was then purified and digested with EcoRI to check the presence of an insert. PICOGREEN assay, HindIII digestion and absorbance were used to quantify the DNA. The purified DNA was then sequenced and searched using different databases to find a match. Finally, the protein of interest was identified to be Sentrin-specific protease 2(SUMO-1 protease 1 ) which processes SUMO precursor into their mature form and also causes deSUMOylation from their target substrate.DOI: http://dx.doi.org/10.3329/ijpls.v2i1.15131 International Journal of Pharmaceutical and Life Sciences Vol.2(1) 2013: 10-21

Lipoprotein lipase (LPL) plays a central role in lipoprotein metabolism by hydrolyzing the core triglycerides (TGs) of circulating chylomicrons and very-low-density lipoprotein (VLDL). The effects of LPL polymorphisms on lipid levels and coronary artery disease (CAD) have been inconsistent among studies and populations. To assess the lipid profiles and distributions of three LPL gene polymorphisms in Saudi patients with CAD, the HindIII, PvuII and Ser447Ter polymorphisms in the LPL gene were analyzed in 226 patients with CAD and 110 controls. Polymerase chain reaction-restriction fragment length polymorphism was used to detect LPL gene polymorphisms. The plasma lipid profiles of the patients were determined using standard enzymatic methods. Patients in the CAD group had significantly higher triglyceride (TG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) levels than controls irrespective of the HindIII, PvuII or Ser447Ter genotype. Compared to the findings in controls, the HindIII TT, PvuII TC and Ser447Ter CC genotypes were associated with significantly reduced high-density lipoprotein cholesterol (HDL-C) levels in patients with CAD (P

The 16 somatic serotype type strains and 60 field isolates of Pasteurella multocida, representing various avian species and geographic regions in Hungary, were characterised by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the ompH gene with DraI restriction endonuclease. The type strains yielded eight different (I-VIII) profiles. Strains whose PCR fragment was uncut by DraI (profile IV) could be differentiated with HindIII and PvuII restriction endonucleases. Five of the eight PCR-RFLP profiles (I, III, V, VI and VII) were detected among the field strains. Only a correlation of limited strength was found between the classical somatic serotypes and the PCR-RFLP profiles. However, the results confirmed that molecular methods could confidently distinguish serotype A:1 strains from the other serotypes. Moreover, the specific relationship between somatic serotypes and PCR-RFLP types among isolates from turkey raises the possibility of the existence of host-specific clones within the P. multocida population.

Protein and DNA polymorphismswere surveyed among seven accessions of wild fenugreek (Trigonellafoenum-graecum L.) to estimate their genetic diversity and relationships. Samples were obtained from diverse ecogeographical areas in Saudi Arabia and Yemen. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of seed storage protein showed genetic variations among fenugreek germplasms, both quantitatively and qualitatively, generating a total of 168 polypeptide bands with different molecular weights ranging from 4.5 to 300 kDa. Twenty-six of these bands were polymorphic, with a considerable polymorphism value (80.00%). Furthermore, restriction fragment length polymorphism (RFLP) analysis was also employed, which was based on the ability of four restriction enzymes (EagI, EcoRI, FspI, and HindIII) to cleave genomic DNA of the plant materials at specific target nucleotide sequences into different numbers of DNA fragments. RFLP analysis revealed 166 fragments with known sequences and variable lengths ranging from 80 to 4000 bp with a highly degree of polymorphism (88.71%). Data derived from SDS-PAGE or RFLP analyses were used to produce dendrograms, which clustered the studied fenugreek accessions into different groups based on the unweighted pair group method with arithmetic mean (UPGMA). The resulting relationships indicated that these two marker techniques were nearly equivalent, but not identical, with respect to phylogenetic information. In conclusion, SDS-PAGE analysis of seed proteins should be augmented with RFLP analysis of DNA for reliable estimates of genetic diversity among fenugreek germplasms.

This study was conducted to detect the genetic variants (single nucleotide polymorphism) of kappa-casein locus (Asp148/Ala) in F1: Simmental (Bos taurus) x Ongole grade (Bos indicus), SIMPO. Genomic DNA was isolated from blood sample of 40 SIMPO (21 males and 19 females). A 780 bp specific fragment of kappa-casein gene spanning from the forth exon region (517 bp) to forth intron (263 bp) was successfully amplified. The result of the PCR-RFLP (Polymerase Chain Reaction – Restriction Fragment Length Polymorphisms) analysis using HindIII enzyme showed that two genotypes (AA and AB) were found at this locus in SIMPO. The frequencies of A and B alleles in SIMPO were 0.79 and 0.21, respectively. The frequency lies between B. taurus (Simmental) and B. indicus group.

A novel method based on (1) initial microbiological screening and (2) a highly specific PCR is described for selection of strains expressing YGNGV motif-containing pediocin. Initial screening is carried out using spot on the lawn assay for selection of acid-free, hydrogen peroxide (H2O2)-free and secreted heat-stable inhibitory activity producing strains. This is followed by highly specific PCR for amplification of 406-bp fragment using forward primer: 5'-tggccaatatcattggtggt-3' targeting signal peptide sequence of pediocin structural gene and reverse primer: 5'-ctactaacgcttggctggca-3' encoding N-terminus of immunity gene. The assay was validated with Pediococcus pentosaceus NCDC273 and Pediococcus acidilactici NCDC252 using (1) digestion of amplified 406-bp fragment with HindIII restriction enzyme-producing two restriction fragments of expected sizes (227 and 179 bp), (2) nucleotide sequencing of 406-bp fragment from both strains found these pediocins identical to pediocin PA-1/AcH and (3) identification of both pediocins as pediocin PA-1 at protein level using RP-HPLC. The assay was used for screening six strains (3 pediococci, 2 lactobacilli and an Enterococcus faecium) producing acid-free, hydrogen peroxide (H2O2)-free and secreted heat-stable inhibitory activity. This resulted in the detection of three new strains (P. pentosaceus NCDC35, E. faecium NCDC124 and Lactobacillus plantarum NCDC20) producing YGNGV motif-containing pediocins.

No epidemiological typing system to differentiate among Pseudomonas pseudomallei isolates has been available. Ribotype analysis was developed and used to examine 74 clinical and 10 environmental isolates of P. pseudomallei from Thailand. Six P. pseudomallei ribotypes were identified from restriction fragment polymorphisms of EcoRI chromosomal digests. The predominant ribotype, A, was found in 59 of the isolates examined. By using patterns from hybridizations with SalI, HindIII, and PstI restriction digests, isolates of ribotype A were subdivided into a further five subtypes, giving a total of 10 differentiable P. pseudomallei types. In 23 of 34 melioidosis patients studied, multiple P. pseudomallei isolates were present. In all but one of these patients, a single ribotype of the organism was present. Isolation of two different ribotypes of P. pseudomallei from one patient, one each in sputum and urine, suggests that superinfection may have occurred. The ribotype was shown to be conserved during the course of antibiotic treatments in seven patients studied, although the antibiotic sensitivity patterns in the isolates from these patients varied. The prevalence of subtype A1 in clinical and environmental specimens suggests that this strain may be predominant in this geographical location. These results demonstrate the usefulness of the ribotyping method for epidemiological studies of P. pseudomallei.