Antimicrobial activity of Lactobacillus pentosus against the Bacillus cereus and Klebsiella pneumoniae strains.

Food safety is the primary concern of the food industry. The aim of the current research is to study the antimicrobial effects of cell-free supernatant of Lactobacillus pentosus against Bacillus cereus and Klebsiella pneumoniae. B. cereus and K. pneumoniae were isolated from infant formula milk product and meat sample, respectively. Their identification was performed through morphological characterization and biochemical testing. Molecular identification of K. pneumoniae was based on 16s ribotyping. A previously isolated and reported strain of L. pentosus was used for the isolation of CFS (Cellfree supernatants). Antimicrobial activity was studied through agar well diffusion assay. Inhibitory activity was recorded by measuring the zone of inhibition. CFS activity was evaluated for temperature and pH. The antimicrobial activity of CFS of L. pentosus produced at different temperatures and pH was investigated against B. cereus and K. pneumoniae. A clear zone of inhibition was observed against B. cereus while no ZOI was formed against K. pneumoniae. K. pneumoniae was found resistant to the CFS. Crude bacteriocin exhibited heat stability for a temperature of 121°C for 30 minutes and pH range of 3-7. The current study concluded that bacteriocin produced from L. pentosus can be used for the control of B. cereus. Its heat and pH stability allows its potential therapeutic use in the food industry as a food preservative and to control food poisoning cases due to B. cereus. K. pneumoniaeis was found resistant to the isolated bacteriocin, and therefore L. pentosus cannot be used for control against K. pneumoniae.

Association of rs1800668 polymorphism in glutathione peroxidase- 1 gene and risk of rheumatoid arthritis in Pakistani population.

OBJECTIVE: To investigate the role of glutathione peroxidase 1 (GPX1) C/T polymorphism (rs1800668) in modulating the chances of Rheumatoid arthritis (RA) in Pakistani population. METHODS: A total of 400 individuals including 200 controls and 200 patients of RA, were genotyped. Detection of rs1800668 polymorphism was carried out using PCR based amplification strategy (allele specific). RESULTS: The results for Hardy Weinberg Equilibrium (HWE) indicated that the allele frequencies for GPX1 polymorphism were not deviant from HWE in whole population under observation. The statistical analysis indicated that significant association existed between rs1800668 polymorphism and RA (p<0.01). CT genotype increased the risk of RA development by 1.8582 times (OR: 1.8582; 95% CI 1.2154 to 2.8409). CC genotype was found to have protective effect against the disease development (OR: 0.5133; 95% CI 0.3403 to 0.7742) while TT genotype was found to have association with RA development but the risk level was marginal (OR: 1.5319; 95% CI 0.6124 to 3.8322). CONCLUSION: The present finding suggests the importance of GPX1 C/T polymorphism (rs1800668) in development of RA in Pakistani population. The protective role of CC genotype against the development of RA in local population was also observed.

Genetic etiology of coronary artery disease considering NOS 3 gene variant rs1799983.

Reduced production of nitric oxide due to rs1799983 single nucleotide polymorphism in nitric oxide synthase 3 gene (NOS3) may enhance the risk of coronary artery disease. The association of rs1799983 polymorphism with coronary artery disease was investigated in the local population of Pakistan. Study consisted of 376 individuals, out of which 198 were coronary artery disease patients and 178 were normal healthy individuals. Allele-specific polymerase chain reaction (PCR) based strategy was used for the detection of different genotypes of rs1799983 polymorphism. PCR amplification results were obtained for 354 samples. Frequency of T allele was higher as compared to G allele in our population. Strong association between rs1799983 and coronary artery disease was observed (p < 0.01). TT genotype was found to enhance 5.717 times the risk of coronary artery disease (odds ratio (OR): 5.717; 95% confidence interval (95% CI) 3.586-9.115). On the basis of present results, it can be concluded that rs1799983 is strongly associated with coronary artery disease in our population and TT genotype of this polymorphism enhanced the risk of coronary artery disease in Pakistani population.

Role of MyD88-Adaptor-Like (MAL) Gene Polymorphism rs8177374 and Cytokine (IFN-γ, TNF-α, IL-10, TGF-ß) Levels in Diverse Malaria Manifestations upon P. falciparum and P. vivax Infections.

This study investigated the role of genetic variant rs8177374 in MAL/TIRAP gene in mediating the cytokine levels of IFN-γ, TNF-α, IL-10, and TGF-ß in malaria patients due to Plasmodium falciparum or P. vivax infection. The study included human blood samples collected from patients with malaria (n = 228) and healthy controls (n = 226). P. falciparum and P. vivax groups were established based on the causative species of Plasmodium. Malaria samples were divided into mild and severe malaria groups based on the symptoms that appeared in the patients, according to the WHO criteria. In a previous study, we genotyped rs8177374 via allele specific PCR strategy. In this study, cytokine levels were estimated in the blood plasma of rs8177374 genotype samples via Sandwich Enzyme Linked Immunosorbent Assay kits. Increased IFN-γ and TNF-α levels in presence of CC genotype indicates the role of CC genotype in both severe and mild malaria groups. Enhanced IL-10 levels in the CT genotype and mild malaria groups suggest a role of CT genotype and IL-10 in the mild clinical outcomes of malaria. The rs8177374 polymorphism in MAL/TIRAP plays an important role in malaria pathogenesis.

Evaluation of the anti-inflammatory, analgesic, anti-pyretic and anti-ulcerogenic potentials of synthetic indole derivatives.

A large number of new synthetic compounds are synthesized in the field of heterocyclic chemistry having a variety of biological potentials. In the present study, some synthetic indole derivatives are used to check anti-inflammatory, analgesic, antipyretic and gastroprotective activity in albino mice. Albino mice of either sex of reproductive age were used for each study (n = 5). In anti-inflammatory activity, the negative control (NC) and positive control group animals were treated with normal saline and 10 mg/kg of indomethacin respectively. The treated groups received the twenty four different synthetic chemicals, after 30 min of sub cutaneous injection of carrageenan. In analgesic activity, hot-plate method is used and for each group the latency period was recorded at zero moment of the provision of required dose and after 30, 60, 90, 120 and 180 min. In anti-pyretic activity, Pyrexia was induced by using Brewer's yeast method. Before any treatment and then after duration of 18 h, the rectal temperatures were recorded. Among all the chemicals, only those chemicals which show any potential related to above mentioned activities were selected for gastroprotective activity. The gastroprotective activity was performed to check the gastric ulcers by using 300 mg/kg of single oral dose of indomethacin to animals of all groups except NC group. This study helped to screen out the most potent indole derivatives 3a-II and 4a-II from the 24 synthetic indole derivatives which demonstrated the best biological potential (anti-inflammatory, analgesic, antipyretic, and gastroprotection) as compared to the remaining ones. The micrometric and biochemical results also support the histological findings. Out of the twenty-four novel indole amines tested, 3a-II and 4a-II have shown the effective pharmacological capacity and additionally have not shown any overt and systemic toxicity. Thus these two indole amines need further in-depth pharmacokinetic and pharmacodynamics studies before they are recommended for any pre-clinical trial.

Association of ACE ID and ACE G2350A polymorphism with increased blood pressure in persons exposed to different sound levels in Pakistan.

PURPOSE: To investigate the role of ACE ID and ACE G2350A polymorphism in increasing the blood pressure of persons exposed to different sound levels. METHOD: Persons exposed to normal sound level (≤80 dBA) and noise (>80 dBA) were divided in two categories on the basis of blood pressure. Category A was comprised of the individuals with blood pressure ranges equal or less than that for normotensive persons. Category B was composed of the workers with blood pressure above than that for category A. ACE ID and ACE G2350A polymorphism was detected for analysis of its association with the increased blood pressure considering noise as modifier. RESULTS: Least homozygous allele model for ACE ID polymorphism indicates no association of DD allele with the increased blood pressure in persons exposed to normal sound (Odds ratio: 0.831; 95% Confidence interval: 0.340-2.030). Similar findings were noted in persons exposed to noise (Odds ratio: 0.992; 95% Confidence interval: 0.536-1.835). Combining all the samples did not change the results (Odds ratio: 0.948; 95% Confidence interval: 0.580-1.549). For ACE G2350A polymorphism, least common homozygote model revealed that AA allele did not affect the chances of increased blood pressure in normal sound exposed group (Odds ratio: 0.827; 95% Confidence interval: 0.169-4.042) and noise exposed group (Odds ratio: 1.416; 95% Confidence interval: 0.682-2.941). There was no prominent variation in the results on combining all the samples (Odds ratio: 1.529; 95% Confidence interval: 0.807-2.872). CONCLUSIONS: No association was observed between ACE ID/ACE G2350A polymorphism and the increased blood pressure in persons exposed to normal sound and noise in Pakistani population.

Antimicrobial efficacy of Lactobacillus plantarum strain against the B. cereus, B. subtilis, S. aureus and E.coli strains

The current study aimed to detect the antimicrobial effect of cell free supernatant (CFS) of Lactobacillus plantarum strain against Gram +ve and Gram ­ve bacterial strains. The strain of Lactobacillus plantarum was isolated using selective media MRS agar. The strain was characterized on the basis of the gram staining, colony morphology, the biochemical tests and the DNA sequencing based method of 16S ribotyping. A total of four test strains (The three already isolated and reported strains (E.coli, S. aureus and B. subtilis) and the one recently identified novel strain (B. cereus) were used for the analysis of antagonistic activity of bacteriocin produced by L. plantarum strain. The CFS of L. plantarum showed zone of inhibition against all the test strains (Gram +ve and Gram ­ve bacteria). The conditions favoring the growth of bacteria were associated with the antimicrobial efficacy of CFS. Bacteriocin activity of CFS remained effective after exposure to temperature stress. Wide range of antagonistic potential of CFS of L. plantarum provides an alternative for antibiotics in pharmaceutical industry. Heat resistant feature of bacteriocin suggests its application in food industry.

Role of rs4986790 Polymorphism of TLR4 Gene in Susceptibility towards Malaria Infection in the Pakistani Population.

BACKGROUND: Toll-like receptors (TLRs) of the human immune mechanism play important role in the detection of invading pathogens. TLRs specifically recognize the pathogen-associated molecular patterns (PAMPs) from pathogens and start the effective response. Single nucleotide polymorphisms (SNPs) in the TLRs can mediate their functions. Present study evaluated the importance of rs4986790 polymorphism of TLR4 gene in susceptibility towards malaria, clinical outcomes of the disease and responsible species of malaria. METHODS: Blood samples of 228 malaria patients and 226 healthy volunteers were selected for the study. Sample collection was completed during Sep 2013 to Sep 2015 from different hospitals of Punjab, Pakistan. Patient's samples were divided into P. vivax group and P. falciparum group on the basis of causative species of Plasmodium. Malaria samples were also divided into mild and severe malaria group based on clinical outcomes of the disease according to WHO criteria. Healthy individuals were placed in the control group. Whole blood was used for the isolation of DNA. Genomic DNA was isolated and amplification of targeted SNP was performed using allele-specific PCR. RESULTS: Results indicate the protective role of AA genotype against the susceptibility of P. vivax infection, OR: 0.5, 95%CI: 0.285-0.876, P=0.038. CONCLUSION: rs4986790 polymorphism of TLR4 gene modulates the susceptibility towards P. vivax infection. AA genotype is found to be protective against the development of P. vivax infection in the local population of Pakistan.

Role of MyD88-adaptor-like gene polymorphism rs8177374 in modulation of malaria severity in the Pakistani population.

INTRODUCTION: The present study was designed to investigate the association between rs8177374 polymorphism and malaria symptoms due to exposure of Plasmodium vivax and Plasmodium falciparum. MATERIALS AND METHODS: A total of 454 samples were included in the study (228 malaria patients and 226 healthy individuals). Malaria patients, divided into P. vivax and P. falciparum groups on the basis of the causative species of Plasmodium, were categorized into mild and severe on the basis of clinical outcomes according to WHO criteria. Healthy individuals were used as controls. Allele specific PCR based strategy was used for the identification of rs8177374 SNP. RESULTS: MyD88-adaptor-like gene polymorphism was associated with susceptibility to malaria (p<0.001). C allele frequency (0.74) was higher in the population compared to T allele frequency (0.26). CT genotype increased the susceptibility of malaria (OR: 2.661; 95% CI: 1.722-4.113) and was positively associated with mild malaria (OR: 5.609; 95% CI: 3.479-9.044, p=0.00). On the other hand, CC genotype was associated with severe malaria (OR: 3.116; 95% CI: 1.560-6.224, p=0.00). P. vivax infection rate was higher in CT genotype carriers compared to other genotypes (OR: 3.616; 95% CI: 2.219-5.894, p<0.001). CONCLUSION: MyD88-adaptor-like/TIR domain containing adaptor protein polymorphism for single nucleotide polymorphism rs8177374 is related with the susceptibility of malaria.

Association between Genetic Polymorphism and Risk of von Willebrand Disease in Pakistan.

von Willebrand disease (VWD) is an inherited, genetically and clinically heterogeneous hemorrhagic disorder. The most common cause of this disease is mutation in the gene that encodes protein von Willebrand factor (VWF) which is responsible for blood clotting. The current study was designed to investigate the role of genetic polymorphisms with the onset of VWD in population of Pakistan. Three exonic variants (c.3445T>C; c.4975C>T; c.7603C>T) from VWF gene were used for the genotyping purpose. The current study employed a case-control association design involving 43 VWD patients and 100 healthy controls from Pakistani population. The genetic reason of VWD was investigated using the allele specific PCR. The significant (P < 0.05) allelic association was found between all three exonic variants and VWD. The CT genotype of these variants was noticed to be associated with significantly higher risk of VWD [odds ratio (95% CI): 14.7 (4.546-47.98), 26.71 (7.281-97.98), and 21.5 (5.806-80.01) for c.3445T>C, c.4975C>T, and c.7603C>T, resp.] while genotypes CC (c.4975C>T) and TT (c.3445T>C and c.7603C>T) were having protective effect against the disease. However, replicated studies are needed for elaborating the role of these SNPs.

Association of the rs10757274 SNP with coronary artery disease in a small group of a Pakistani population.

OBJECTIVE: The present study aimed to investigate the association between the rs10757274 SNP (present on locus 9p21 in the gene for CDKN2B-AS1) and coronary artery disease (CAD) in a local population of Pakistan. METHODS: It was a case-control study. An allele-specific PCR-based strategy was used for the identification of genotypes. A total of 350 samples were used for the investigation, out of which 220 samples were CAD patients and 130 samples were normal healthy individuals. Effects of parameters, like family history of CAD, smoking, presence of diabetes, and hypertension, in changing the chances of CAD were studied. Odds ratio was estimated with 95% confidence interval. RESULTS: A strong association was observed between CAD and factors, like smoking (OR: 1.666; 95% CI: 1.042-2.664), presence of hypertension (OR: 26.55; 95% CI: 15.95-44.20), diabetes (OR: 3.009; 95% CI: 1.841-4.920), and family history of CAD (OR: 4.9; 95% CI: 2.965-8.099). Results for the association between the genotype on the basis of rs10757274 showed a strong association between the GG genotype and the occurrence of CAD (OR: 9.603; 95% CI: 5.746-16.05). CONCLUSION: The present results suggest the importance of the 9p21 locus in modulating the chances of CAD.

Role of S180L polymorphism in etiology of malaria caused by Plasmodium falciparum in a small group of Pakistani population.

The aim of our study was to investigate the role of S180L polymorphism in modulation of acquisition of malaria caused by Plasmodium falciparum in a small group of Pakistani population. A total of 133 individuals including 60 controls and 73 patients of malaria, caused by Plasmodium falciparum, were genotyped using allele-specific PCR. Ninety-two samples successfully demonstrated the PCR amplification results, while forty-one samples could not be genotyped due to failure in PCR amplification. The allele frequency for S180L polymorphism was deviant from Hardy-Weinberg equilibrium (HWE) of the population under observation. Association was found between the observed polymorphism and the occurrence of malaria caused by Plasmodium falciparum (p = 0.01). Chances of malaria caused by Plasmodium falciparum were low in CC genotype carriers in comparison to other genotypes (Odds ratio: 0.3016; 95% CI: 0.124-0.729). The present findings suggest that S180L polymorphism is important in modulating the probability of acquisition of malaria caused by Plasmodium falciparum in Pakistani population. The CC genotype plays a protective role in local population against this type of malaria.

Electrolyte Disturbance and the Type of Malarial Infection.

BACKGROUND: Electrolytes play an important role in the normal functioning of human body. Electrolyte imbalance and mineral disturbances is the common clinical manifestation in several infectious diseases including malaria. Malaria is a mosquito borne serious infectious disease of the world. Plasmodium vivax and P. falciparum are the main agents responsible for malaria in Pakistan. Electrolyte imbalance in malarial infection may lead towards the severity of disease. METHODS: The present study analyzed the electrolytes levels (Na, K, Ca and Mg) in malarial patients and healthy individuals. Patients were categorized into two groups, P. falciparum and P. vivax, based on causative species of Plasmodium. Study consisted of 173 individuals, out of which 73 were malarial patients and 100 were normal healthy individuals. RESULTS: Concentrations of Na, K, and Ca were low in the blood of malarial patients as compared to healthy individuals (P<0.05). No significant difference for these electrolytes exists between P. falciparum and P. vivax infected groups (P>0.05). The concentration of Mg was changed based on exposure to the type of parasite. In P. falciparum infection, the level of Mg was lower than healthy individuals was (P<0.05). Discordantly, in case of P. vivaxinfection, Mg level was higher than healthy individuals were (P<0.05). No variation was noticed in electrolytes levels due to gender differences (P>0.05). CONCLUSION: Variation in Mg levels occurs due to exposure of Plasmodium depending on its type. The levels of Na, K and Ca are also changed due to Plasmodium, regardless of its type.

Role of MyD88-adaptor-like gene polymorphism rs8177374 in modulation of malaria severity in the Pakistani population

Abstract Introduction: The present study was designed to investigate the association between rs8177374 polymorphism and malaria symptoms due to exposure of Plasmodium vivax and Plasmodium falciparum. Materials and methods: A total of 454 samples were included in the study (228 malaria patients and 226 healthy individuals). Malaria patients, divided into P. vivax and P. falciparum groups on the basis of the causative species of Plasmodium, were categorized into mild and severe on the basis of clinical outcomes according to WHO criteria. Healthy individuals were used as controls. Allele specific PCR based strategy was used for the identification of rs8177374 SNP. Results: MyD88-adaptor-like gene polymorphism was associated with susceptibility to malaria (p < 0.001). C allele frequency (0.74) was higher in the population compared to T allele frequency (0.26). CT genotype increased the susceptibility of malaria (OR: 2.661; 95% CI: 1.722-4.113) and was positively associated with mild malaria (OR: 5.609; 95% CI: 3.479-9.044, p = 0.00). On the other hand, CC genotype was associated with severe malaria (OR: 3.116; 95% CI: 1.560-6.224, p = 0.00). P. vivax infection rate was higher in CT genotype carriers compared to other genotypes (OR: 3.616; 95% CI: 2.219-5.894, p < 0.001). Conclusion: MyD88-adaptor-like/TIR domain containing adaptor protein polymorphism for single nucleotide polymorphism rs8177374 is related with the susceptibility of malaria.

Effects of noise exposure on catalase activity of growing lymphocytes.

Oxidative stress due to noise was estimated at cell level using model of growing lymphocytes. Lymphocytes were isolated and cultured using conventional methodology. Cell culture of each group was exposed to sound of frequency 1 KHz during incubation. Three groups were defined on the basis of exposure of sound with specific range of intensity and duration of exposure. Group A and Group B were exposed to sound with intensity 110 dBA for four hours per day and for eight hours per day respectively. Control group was exposed to sound less than 85 dBA. Viable cell count was performed using trypan blue. Catalase activity of each group was estimated using ELISA kit.Viable cell count of Group A and Group B was almost same but significantly less than that of control group. Catalase activity of lymphocytes in Group B was significantly low as compared to Group A and controls (p=0.003, p< 0.05). There was no significant difference between catalase activity of Group A and control group.Exposure of sound with frequency 1 KHz and intensity 110 dBA for 4 hours and eight hours per day may induce oxidative stress in growing lymphocytes causing the difference in viable cell count. However the catalase activity depends on duration of exposure. In case of noise exposure of 8 hours per day, it declines significantly as compared to noise exposure of 4 hours per day.

Effect of ACE polymorphisms on the association between noise and hypertension in a Pakistani population.

INTRODUCTION: This study investigates the effect of angiotensin converting enzyme (ACE) polymorphisms on the association between noise and hypertension. SUBJECTS AND METHODS: A cross sectional study was conducted involving 385 male volunteers. In the control group 156 participants were exposed to a sound level of ≤ 80 dB; in the exposed group 229 individuals were exposed to a sound level ≥ 80 dB. Both groups were subdivided into categories on the basis of blood pressure. A polymerase chain reaction (PCR)-based strategy was used to differentiate the various genotypes of ACE ID and ACE G2350A. Association of noise with genotypes as modifiers was estimated after adjustment of data for age and body mass index. RESULTS: These showed the following odds ratios (OR) with 95% confidence interval (CI) indicating that noise is associated with hypertension in a Pakistani population (OR: 3.196; CI: 1.803-5.666). ACE ID and ACE G2350A polymorphism affects this association. ID (OR: 2.844: CI: 1.32-6.110) and DD allele carriers (OR: 4.487; CI: 1.549-12.99) were at increased risk of hypertension. GG (OR: 3.970; CI: 1.628-9.681) and GA (OR: 3.705; CI: 1.407-9.758) carriers were also at increased risk of hypertension associated with noise. However II and AA allele carriers were not found to be at increased risk of hypertension. CONCLUSION: ACE ID polymorphism and ACE G2350A polymorphism modulates the effects of noise on blood pressure.

Noise induced hypertension and prehypertension in Pakistan.

The present study investigates the relationship of different sound levels with hypertension and prehypertension in Pakistani population. A cross sectional study was conducted to determine the prevalence of hypertension and prehypertension due to exposure of sound level ≤80 dBA (A weighted sound pressure level), 81-94 dBA and ≥95 dBA in November 11, 2005 to January 30, 2007. Sites were selected with stable sound ranges according to the above mentioned criteria. After selecting sampling sites, workers living in that area for at least 8 hours per day were categorized on the basis of blood pressure in groups called as normotensive, prehypertensive and hypertensive. Persons with diabetes, chronic bacterial or viral infections, alcohol addiction, kidney problems were excluded from the study. For getting homogenous groups, age range of 30-50 years was selected. Out of 566 samples, 90 excluded samples were consisted of 8% diabetic patients, 5% hepatitis C patients, 3% hepatitis B patients and 0% AIDS patients. Out of 476 participants, 389 samples were found with age 40±10 years. High noise increased the risks of hypertension (Odds ratio: 4.41; Confidence interval: 2,123-9,196) and prehypertension (Odds ratio: 3,809; Confidence Interval: 1,804-8,042) as compared to the normal sound level. However increased chances of hypertension (Odds ratio: 2,271; Confidence interval: 1,043-4,946) and prehypertension (Odds ratio: 3,028; Confidence Interval: 1,440-6,367) were observed on median noise exposure also. These findings suggest that sound level more than 81 dBA increases the chances for development of hypertension and prehypertension in Pakistani population.

Bacteriocins produced by L fermentum and L acidophilus can inhibit cephalosporin resistant E coli

Reemerging infections occur due to resistant bacteria. Such infections create restrictions for clinicians and microbiologists in drug selection. Such problems demand new strategies for solution. Use of bacteriocins for this purpose may be fruitful. In the present research work, the inhibitory effects of bactericins on cephalosporin resistant Escherichia coli are used as model system for the control of antibiotic resistant pathogenic bacteria. Cephalosporin resistant Escherichia coli strain was isolated from pus by using conventional methodology. For bacteriocin production, Lactobacilli strains were selected by using selective media. Out of seventy two strains isolated from yogurt, fecal materials of human, chick, parrot and cat, only two strains (strain 45 and strain 52) were found to produce bacteriocins having antimicrobial potential against cephalosporin resistant Escherichia coli. Biochemical characterization showed that strain 45 belonged to group of Lactobacillus fermentum and strain 52 to Lactobacillus acidophilus. Both strains showed maximum growth at 25ºC and 35ºC respectively. Suitable pH was 5.5 and 6.0 for Lactobacillus fermentum and Lactobacillus acidophilus respectively. Bacteriocins produced by both strains were found stable at 50, 75 and 100ºC for 60min. Function of bacteriocin was also not disturbed due to change in pH. These findings suggest that bacteriocin produced by Lactobacillus fermentum and Lactobacillus acidophilus can be used for the infection control of cephalosporin resistant Escherichia coli.