Computer controlled expansion of equine cord blood mesenchymal stromal cells on microcarriers in 3 L vertical-wheel® bioreactors.

Mesenchymal stromal cells (MSCs) are an ideal cell source for allogenic cell therapy due to their immunomodulatory and differentiation properties. Equine MSCs (eMSCs) have been found to be a promising treatment for equine joint injuries including meniscal injuries, cartilage degradation, and osteoarthritis. Although the use of eMSCs has shown efficacy in preliminary studies, challenges associated with biomanufacturing remain. To achieve the required cell numbers for clinical application, bioreactor-based processes are required. Initial studies have shown that eMSCs can be cultivated in microcarrier-based, stirred suspension bioreactor culture at the laboratory 0.1 L scale using a Vertical-Wheel® (VW) bioreactor. However, investigations regarding scale up of these processes to the required biomanufacturing scales are required. This study investigated the scale-up of a equine cord blood MSC (eCB-MSC) bioprocess in VW bioreactors at three scales. This included scale-up from the 0.1-0.5 L bioreactor, scale-up from static culture to the 3 L computer-controlled bioreactor, and scale-up into the 3 L computer-controlled bioreactor using a mock clinical trial process. Results from the various scale-up experiments demonstrated similar cell expansion at the various tested scales. The 3 L computer-controlled system resulted in a final cell densities of 1.5 × 105 cells/cm2 on average, achieving 1.5 × 109 harvested cells. Biological testing of the cells showed that cell phenotype and functionality were maintained after scale-up. These findings demonstrate the scalability of an eCB-MSC bioprocess using microcarriers in VW bioreactors to achieve clinically relevant cell numbers, a critical step to translate MSC treatments from research to clinical applications. This study also represents the first known published study expanding any cell type in the 3 L VW bioreactor.

Association of diverse bacterial communities in human bile samples with biliary tract disorders: a survey using culture and polymerase chain reaction-denaturing gradient gel electrophoresis methods.

Bacterial infection is considered a predisposing factor for disorders of the biliary tract. This study aimed to determine the diversity of bacterial communities in bile samples and their involvement in the occurrence of biliary tract diseases. A total of 102 bile samples were collected during endoscopic retrograde cholangiopancreatography (ERCP). Characterization of bacteria was done using culture and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) methods. Antimicrobial susceptibility of the isolates was determined based on the Clinical and Laboratory Standards Institute (CLSI) guidelines and identity of the nucleotide sequences of differentiated bands from the DGGE gels was determined based on GenBank data. In total, 41.2 % (42/102) of the patients showed bacterial infection in their bile samples. This infection was detected in 21 % (4/19), 45.4 % (5/11), 53.5 % (15/28), and 54.5 % (24/44) of patients with common bile duct stone, microlithiasis, malignancy, and gallbladder stone, respectively. Escherichia coli showed a significant association with gallstones. Polymicrobial infection was detected in 48 % of the patients. While results of the culture method established coexistence of biofilm-forming bacteria (Pseudomonas aeruginosa, E. coli, Klebsiella pneumoniae, Enterococcus spp., and Acinetobacter spp.) in different combinations, the presence of Capnocytophaga spp., Lactococcus spp., Bacillus spp., Staphylococcus haemolyticus, Enterobacter or Citrobacter spp., Morganella spp., Salmonella spp., and Helicobacter pylori was also characterized in these samples by the PCR-DGGE method. Multidrug resistance phenotypes (87.5 %) and resistance to third- and fourth-generation cephalosporins and quinolones were common in these strains, which could evolve through their selection by bile components. Ability for biofilm formation seems to be a need for polymicrobial infection in this organ.

Seroprevalence of Helicobacter pylori in Nahavand: a population-based study.

In this cross-sectional study, we evaluated H. pylori seroprevalence and the relevant factors in 1518 people aged > or = 6 years from the general population of Nahavand, western Islamic Republic of Iran. Questionnaires covering sociodemographic variables were completed by interview. Blood samples were taken from each individual. Sera were tested for anti-H. pylori IgG using commercial enzyme immunoassay. Overall, seroprevalence of H. pylori was high, 71.0% (95% CI: 69.0%-73.0%). There was a gradual increase with age. Based on multivariate adjustment, only female sex and age could be considered risk factors.

Seroprevalence of hepatitis E in Nahavand, Islamic Republic of Iran: a population-based study.

A 2-month community-based survey in February/March of 2003 was carried out to study the seroprevalence of hepatitis E virus (HEV) infection in Nahavand, Islamic Republic of Iran. From each of 6 urban regions of Nahavand, 304 inhabitants > or = 6 years were recruited through systematic random sampling (total 1824). Participants were tested for anti-HEV IgG using ELISA. The overall seroprevalence of HEV was 9.3% (95% CI: 8.2%-10.9%). Based on multivariate adjustment, only sex (OR = 1.61, 95% CI: 1.13-2.28) and age (OR = 1.03, 95% CI: 1.01-1.04) emerged as significant risk factors. This intermediate prevalence urges further investigations on HEV infection in the Islamic Republic of Iran.

Patient concerns regarding chronic hepatitis B and C infection.

Counselling of patients with viral hepatitis is often limited to discussions about how the virus is transmitted. The aim of the present study was to document the principal concerns of patients suffering from chronic hepatitis B and C infection. The most common volunteered concerns were infecting family members (80.6%), infecting others (66.7%), side-effects of treatment (50.0%), disease progression to cirrhosis (44.4%), loss of employment (41.7%), liver transplantation (36.1%), social stigma (36.1%), change in lifestyle (33.3%), development of liver cancer (25%), costs and money (22.2%), fear of disease (5.6%) and psychological disease (5.6%). Health care providers who focus counselling efforts exclusively on viral transmission are unlikely to address other important concerns.

Escherichia coli, Shigella and Salmonella species in acute diarrhoea in Hamedan, Islamic Republic of Iran.

This study investigated the frequency of Escherichia col, Shigella and Salmonella species in stool specimens from patients with diarrhoea presenting to health centres in Hamedan province, Islamic Republic of Iran. From 144 samples, Shigella strains were isolated in 17 cases (11.8%): 10 Sh. flexneri, 3 Sh. sonnei, 2 Sh. boydii and 2 untyped strains. No Salmonella strains were isolated. Using molecular diagnostic methods, diarrheogenic E. coli were detected in 37 cases (25.7%), the majority were enterotoxigenic (ETEC) (22 cases) and Shiga toxin-producing (STEC) strains (15 cases). In 14 cases (9.7%) there was co-infection.

Tumor necrosis factor-alpha gene promoter polymorphism in Iranian patients with chronic hepatitis B.

BACKGROUND: Clearance of hepatitis B virus (HBV) infection requires a good host immune response. Cytokines like tumor necrosis factor-alpha (TNF- alpha) may play a role in such immune response. Genetic changes in TNF-a gene promoter region are known to influence TNF- alpha expression. We therefore studied the role of one such mutation in chronic HBV infection. METHODS: Presence of -308 G/A polymorphism in the promoter region of TNF- alpha gene was looked for in 100 patients with chronic HBV infection, 91 subjects with spontaneously recovered HBV infection and 89 healthy controls, using a PCR-RFLP method. RESULTS: Variant alleles (A/A or A/G) were found in 22 of 100 (22%) patients with chronic HBV infection, 21 of 91 (23%) subjects with spontaneous HBV clearance and 14 of 89 (15.7%) control subjects (p=ns for inter-group comparisons). CONCLUSION: TNF- alpha promoter polymorphism -308A is common in Iranian population, but has no association with development of chronic HBV infection.

Seroprevalence of hepatitis B in Nahavand, Islamic Republic of Iran.

We determined the seroprevalence of hepatitis B in Nahavand in a sample of 1824 subjects > 5 years in 2002. Face-to-face interviews were conducted and blood samples taken. The association between risk factor and hepatitis B was assessed using logistic regression. The prevalence of HBsAg positive cases was 2.3%, and HBcAb and HBsAb were isolated in 7.8% and 11.6% of the participants respectively; 11.9% were positive for both HBcAb and HBsAb. History of surgery and imprisonment were the major risk factors for infection with odds ratios of 2.14 (95% CI: 1.22-3.05) and 3.57 (95% CI: 1.68-5.4) respectively.