Study of Some Virulence Factors of Proteus mirabilis Isolated from Urinary Stones Patients

A total of 125 specimens of stones and urine were collected from urinary stone patients from (June to December, 2012). According to primary identification, which based on macroscopic and microscopic characteristics and biochemical tests, 25 (20%) and 100 (80%) of isolates were identified as Proteus and non-Proteus, respectively. The 25 Proteus isolates were finally identified as Proteus mirabilis based on Vitek 2 system and polymerase chain reaction (PCR) technique by using target gene 16S rRNA. The multiple logistic regressions results showed that the age ˃ 40 years old was a risk factors that significantly associated with increased incidence of P. mirabilis in urinary stone patients, as P = 0.02, and Odd’s ratio (OR) was 4.889 (1.7-14.057), while in relation to gender, the analysis revealed that they were statistically non-significant as OR was 1.174 (0.488-2.822) as well as P=0.720. Some virulence factors of all isolates were investigated by Qualitative and Quantitative assay. Qualitative assay showed that all isolates (100%) were positive to urease, biofilm, Adhesion factors, and swarming activity. Whereas, 40% of isolates were positive to protease and ESBL, 96% and 76% of isolates were positive to agglutination and β-lactamase, respectively. Quantitative assay revealed that all tested isolates were significantly differences P˂ 0.05 in the production of the tested virulence factors. The urease production activity range from 59 to 129 U/ml, whereas the protease production activity ranged from 2.5 to 8 U/ml; the mean of adhering to uroepithelial cell ranged from (20-45) bacteria/cell. The mean of optical density ranged from (0.028-033) at OD630 and the percentages of biofilm activity were 60%, 24% and 16% as strong, moderate and weak biofilm, respectively. The mean of swarming growth activity of P. mirabilis isolates ranged from (3-67mm) and 40%, 32%, 8%, and 20% of isolates showed strong, moderate, weak, and very weak swarming activity, respectively.

The Role of Lipopolysaccharide and O-Antigen of Proteus Mirabilis in Urinary Stones Formation, In-Vitro Study

The ability to induce stone formation in-vitro of Proteus mirabilis isolates was investigated using whole bacterial cell, lipopolysaccharide, and O-antigen of these isolates. The results showed that all three parts have the ability to form crystallization in artificial urine solution, which based on the differences in urease activity and chemical structure of LPS and O-antigen. The whole bacterial cell of isolate No. 6 (P. mirabilis O18) revealed significant differences (P˂ 0.05) in the ability to bind with calcium (251.5 µg/ml) and magnesium (75.4 µg/ml) compared with the whole bacterial cell of isolate No. 14 (P. mirabilis O3) and isolate No. 3 (P. mirabilis O16), where the calcium concentrations were 238.77 µg/ml and 227.12 µg/ml, respectively; and magnesium concentrations were 53.34 µg/ml and 37.68 µg/ml for isolates No. 14 and 3, respectively. In contrast, LPS and O-antigen of isolates No. 14 and 3 showed significantly differences (P˂ 0.05) in the ability to metal binding with calcium (25.77 µg/ml and 25.06 µg/ml), respectively, and magnesium (6.6 µg/ml and 7.5 µg/ml), respectively, compared with LPS and O-antigen of isolate No. 6, where the calcium and magnesium concentration were 18.03 µg/ml and 3.16 µg/ml, respectively

In vivo antibacterial activity of whey protein derived from fermented milk of Iraqi buffalo

The present study aims to prepare fermented buffalo's milk rich with low molecular weight peptides by using lactic acid starters as a mixture. Skim milk sample was inoculated with 5% of the starter. The growing number of starter and anti-bacterial activity were studied after 24 hours of incubation. Protein and peptide concentration were determined before and after fermentation, then biological active peptides were isolated or separated and purified by gel filtration column of Sephadex G25. Finally, the antibacterial activity of the isolated peptides was studied in vivo. The results of chemical analysis of fresh and fermented milk showed that the concentrations of protein were 0.817mg/ml and 0.501mg/ml before and after fermentation, respectively either peptide concentration was 0.4mg/ml before fermentation and 0.805mg/ml after fermentation. The number of starters was determined during the fermentation process after 6, 12, 18 and 24 hours of incubation and found an increase in the number of lactic acid bacteria. The initiation number was 6.2 × 105 but after the 24 hours, the number increased of up to 1.3×106. The number of lactic bacteria decreased after 24 hours with the increase in the concentration of lactic acid combined with low pH value. Colonies of lactobacilli were isolated from fermented buffalo milk and were characterized by the typical characteristics for the purpose of a rating based on morphological and cultural characters. Gel filtration gave seventy-eight fractions. And depending on the absorbency on wavelength 280 were obtained four peaks, each peak represents a fraction. Peptide concentration was determined in each fraction, these concentrations were (0 and 0243 and 0902 and 0632) mg/ml of fraction 1, 2, 3 and 4, respectively. Fraction three contained a high concentration of peptide. The antibacterial activity of the third fraction was estimated. The results showed that the bioactive peptides of fermented milk have good efficacy in the treatment of diarrhea in laboratory animals

Repeatability and Diurnal Variation of Tear Ferning Test

This is a non-final version of an article published in final form in: Masmali, A. M., Al-Bahlal, J. M., El-Hiti, G. A., Akhtar, S., Purslow, C., Murphy, P. J., & Almubrad, T. (2015). Repeatability and Diurnal Variation of Tear Ferning Test: Eye & Contact Lens: Science & Clinical Practice, 41(5), 262–267. https://doi.org/10.1097/ICL.0000000000000116Objectives: To investigate tear ferning (TF) test repeatability between sessions by observing changes in the tear fern pattern during the day. Methods: Twenty-three healthy young adults (15 men and 8 women), ranging in age from 20 to 32 years (mean±SD: 22.9±3.3 years) without signs or symptoms of dry eye disease, ocular disease, or contact lens wear were enrolled in the study. Schirmer I, tear break-up time (TBUT) test, and McMonnies questionnaire were used to screen volunteers. Schirmer I and TBUT tests were applied to both eyes in each subject. Four samples of tear fluid were collected from the right eye of each subject using glass capillaries at set intervals during a single day (9 AM, 11 AM, 2 PM, and 4 PM). The TF patterns obtained from samples were classified according to the Masmali TF grading scale to increments of 0.1. Results: The median values obtained from the McMonnies, Schirmer, and TBUT tests were 4.0±2.0, 30.0±7.0 mm (OD), and 16.0±10.0 sec (OD), respectively. There were no statistically significant differences between the TF grades for tear samples collected at different times of the day (Wilks' Lambda, P=0.351). The majority (84.8%) of TF grades was between 0.0 and 1.5; the remaining 15.2% of subjects had TF between grades 1.6 and 1.9. The overall mean grade for the TF was 1.1±0.3. There were small insignificant correlations between TF grades and the McMonnies questionnaire (r=0.1.30) and TBUT (r=0.248) and a negligible correlation with Schirmer test (r=−0.046). Conclusions: The results found no significant differences within the TF for tear samples collected at different times of the day, suggesting that there is little diurnal variation evident.The authors extend their appreciation to the College of Applied Medical Sciences Research Center and the Deanship of Scientific Research at King Saud University for its funding of this research

Demystifying Smoker's Paradox: A Propensity Score-Weighted Analysis in Patients Hospitalized With Acute Heart Failure.

Background Smoker's paradox has been observed with several vascular disorders, yet there are limited data in patients with acute heart failure (HF). We examined the effects of smoking in patients with acute HF using data from a large multicenter registry. The objective was to determine if the design and analytic approach could explain the smoker's paradox in acute HF mortality. Methods and Results The data were sourced from the acute HF registry (Gulf CARE [Gulf Acute Heart Failure Registry]), a multicenter registry that recruited patients over 10 months admitted with a diagnosis of acute HF from 47 hospitals in 7 Middle Eastern countries. The association between smoking and mortality (in hospital) was examined using covariate adjustment, making use of mortality risk factors. A parallel analysis was performed using covariate balancing through propensity scores. Of 5005 patients hospitalized with acute HF, 1103 (22%) were current smokers. The in-hospital mortality rates were significantly lower in current smoker's before (odds ratio, 0.71; 95% CI, 0.52-0.96) and more so after (odds ratio, 0.47; 95% CI, 0.31-0.70) covariate adjustment. With the propensity score-derived covariate balance, the smoking effect became much less certain (odds ratio, 0.63; 95% CI, 0.36-1.11). Conclusions The current study illustrates the fact that the smoker's paradox is likely to be a result of residual confounding as covariate adjustment may not resolve this if there are many competing prognostic confounders. In this situation, propensity score methods for covariate balancing seem preferable. Clinical Trial Registration URL: https://www.clinicaltrials.gov/. Unique identifier: NCT01467973.Gulf CARE (Gulf Acute Heart Failure Registry) is an investigator- initiated study conducted under the auspices of the Gulf Heart Association and funded by Servier, Paris, France; and (for centers in Saudi Arabia), by the Saudi Heart Association (The Deanship of Scientific Research at King Saud University, Riyadh, Saudi Arabia [research group number: RG -1436- 013]). This does not alter our adherence to policies on sharing data and materials; and the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The publication of this article was funded by the Qatar National Library

Self-reported health status of older adults in Malaysia and Singapore: evidence from the 2007 Global Ageing Survey

The aim of this study is to investigate the correlates of self-reported health (SRH) among older adults in Malaysia and Singapore. The study uses data collected in the Global Ageing Study (GLAS) 2007, one of the largest surveys of its kind, specially designed to investigate attitudes towards later life, ageing and retirement. Data were collected from 1002 and 1004 respondents from Malaysia and Singapore respectively. The study found that Singaporeans report a healthier life than Malaysians. The two countries have consistent results with regard to the influences of selected covariates on individual health. Poorer health is more prevalent among people with lower education, among those widowed, divorced or separated, and those working in blue-collar occupations. Although social support is found to be an important determinant of SRH, the effects are partially confounded with other covariates. These findings enhance our knowledge about the health status of older people, and in turn will be useful for governments to ensure effective policy making

Farnesoid X Receptor Induces Murine Scavenger Receptor Class B Type I via Intron Binding

Farnesoid X receptor (FXR) is a nuclear receptor and a key regulator of liver cholesterol and triglyceride homeostasis. Scavenger receptor class B type I (SR-BI) is critical for reverse cholesterol transport (RCT) by transporting high-density lipoprotein (HDL) into liver. FXR induces SR-BI, however, the underlying molecular mechanism of this induction is not known. The current study confirmed induction of SR-BI mRNA by activated FXR in mouse livers, a human hepatoma cell line, and primary human hepatocytes. Genome-wide FXR binding analysis in mouse livers identified 4 putative FXR response elements in the form of inverse repeat separated by one nucleotide (IR1) at the first intron and 1 IR1 at the downstream of the mouse Sr-bi gene. ChIP-qPCR analysis revealed FXR binding to only the intronic IR1s, but not the downstream one. Luciferase assays and site-directed mutagenesis further showed that 3 out of 4 IR1s were able to activate gene transcription. A 16-week high-fat diet (HFD) feeding in mice increased hepatic Sr-bi gene expression in a FXR-dependent manner. In addition, FXR bound to the 3 bona fide IR1s in vivo, which was increased following HFD feeding. Serum total and HDL cholesterol levels were increased in FXR knockout mice fed the HFD, compared to wild-type mice. In conclusion, the Sr-bi/SR-BI gene is confirmed as a FXR target gene in both mice and humans, and at least in mice, induction of Sr-bi by FXR is via binding to intronic IR1s. This study suggests that FXR may serve as a promising molecular target for increasing reverse cholesterol transport

High Density Lipoprotein (HDL) Promotes Glucose Uptake in Adipocytes and Glycogen Synthesis in Muscle Cells

Background: High density lipoprotein (HDL) was reported to decrease plasma glucose and promote insulin secretion in type 2 diabetes patients. This investigation was designed to determine the effects and mechanisms of HDL on glucose uptake in adipocytes and glycogen synthesis in muscle cells. Methods and Results: Actions of HDL on glucose uptake and GLUT4 translocation were assessed with 1- [ 3 H]-2deoxyglucose and plasma membrane lawn, respectively, in 3T3-L1 adipocytes. Glycogen analysis was performed with amyloglucosidase and glucose oxidase-peroxidase methods in normal and palmitate-treated L6 cells. Small interfering RNA was used to observe role of scavenger receptor type I (SR-BI) in glucose uptake of HDL. Corresponding signaling molecules were detected by immunoblotting. HDL stimulated glucose uptake in a time- and concentration-dependent manner in 3T3-L1 adipocytes. GLUT4 translocation was significantly increased by HDL. Glycogen deposition got enhanced in L6 muscle cells paralleling with elevated glycogen synthase kinase3 (GSK3) phosphorylation. Meanwhile, increased phosphorylations of Akt-Ser473 and AMP activated protein kinase (AMPK) a were detected in 3T3-L1 adipocytes. Glucose uptake and Akt-Ser473 activation but not AMPK-a were diminished in SR-BI knock-down 3T3-L1 cells. Conclusions: HDL stimulates glucose uptake in 3T3-L1 adipocytes through enhancing GLUT4 translocation by mechanisms involving PI3K/Akt via SR-BI and AMPK signaling pathways, and increases glycogen deposition in L6 muscle cells throug

Vitamin D supplementation for the prevention of type 2 diabetes in overweight adults: study protocol for a randomized controlled trial

Despite Australia's sunny climate, low vitamin D levels are increasingly prevalent. Sun exposure is limited by long working hours, an increase in time spent indoors, and sun protection practices, and there is limited dietary vitamin D fortification. While the importance of vitamin D for bone mineralization is well known, its role as a protective agent against chronic diseases, such as type 2 diabetes and cardiovascular disease, is less understood. Observational and limited intervention studies suggest that vitamin D might improve insulin sensitivity and secretion, mainly via its anti-inflammatory properties, thereby decreasing the risk of development and progression of type 2 diabetes. The primary aim of this trial is to investigate whether improved plasma concentrations of 25-hydroxyvitamin D (25(OH)D), obtained through vitamin D supplementation, will increase insulin sensitivity and insulin secretion. A secondary aim is to determine whether these relationships are mediated by a reduction in underlying subclinical inflammation associated with obesity.Fifty overweight but otherwise healthy nondiabetic adults between 18 and 60 years old, with low vitamin D levels (25(OH)D < 50 nmol/l), will be randomly assigned to intervention or placebo. At baseline, participants will undergo a medical review and anthropometric measurements, including dual X-ray absorptiometry, an intravenous glucose tolerance test, muscle and fat biopsies, a hyperinsulinemic euglycemic clamp, and questionnaires assessing diet, physical activity, sun exposure, back and knee pain, and depression. The intervention group will receive a first dose of 100,000 IU followed by 4,000 IU vitamin D (cholecalciferol) daily, while the placebo group will receive apparently identical capsules, both for a period of 16 weeks. All measurements will be repeated at follow-up, with the primary outcome measure expressed as a change from baseline in insulin sensitivity and secretion for the intervention group compared with the placebo group. Secondary outcome measures will compare changes in anthropometry, cardiovascular risk factors, and inflammatory markers.The trial will provide much needed clinical evidence on the impact of vitamin D supplementation on insulin resistance and secretion and its underlying mechanisms, which are relevant for the prevention and management of type 2 diabetes.Clinicaltrials.gov ID: NCT02112721 .Barbora de Courten, Aya Mousa, Negar Naderpoor, Helena Teede, Maximilian P J de Courten and Robert Scrag