Expression of infectious bovine rhinotracheitis virus glycoprotein D in bacterial cell

Bovine Herpesvirus 1 (BHV-1) belongs to the genus of Varicellovirus and the family of Herpesviridae which contains three main gB, gC and gD genes. In order to cloning of the coding region of gD gene of IBR virus , PCR product of the open reading frame of the gene from IBR virus isolated in Iran was amplified by PCR. A 1047bp PCR product of the gD gene with EcoRI, HindIII restriction sites were subcloned of pTZ57R/T and digested by the mentioned endonucleases. Digested insert cloned in to pET-32a and transfered in E.coli cells. For the expression of gD protein, the pET-32a recombinant vector was transformed and then induced in BL21 (DE3) strain of E.coli competent cells using IPTG. The presence of gD expressed protein was shown in immunoblotting and SDS-PAGE system. With respect to the remarkable frequency of infection to IBR in Iran and the necessity of controlling it through vaccination with recombinant vaccines of thymidine kinase, manufacturing and applying the recombinant gD protein are vital goals in recognition and distinction between infection and responsescaused by vaccine

Role of Class I, II and III Integrons in Multidrug Resistance in Pseudomonas aeruginosa Isolated from Nosocomial Hospital

Abstract Aims: The increasing usage of antibiotics can cause resistance to the treatment of infections, which can caused by bacteria, e.g. Pseudomonas aeruginosa. The aim of this study was to trace the class I, II and III integrons in isolates of P. aeruginosa of nosocomial infection and determining the antibiotic resistance pattern of the bacteria. Instrument & Methods: In this cross-sectional study, 100 Pseudomonas aeruginosa clinical isolates of infected wounds, bedsores, burns, urinary tract infections and respiratory tract infections were collected from patients of 3 Isfahan City hospitals, Iran (Al Zahra, Kashani, Shariati) in 2015. After identification tests and antibiogram, integrons class I, II and III were detected by M-PCR method. Data analysis were performed in SPSS 16 software using Chi-square and Fisher exact tests and the relationship between the presence of class III, II, I was calculated by M-PCR test. Findings: All isolates had multiple antibiotic resistances. The highest antibiotic resistance was to Tetracycline (85) and the lowest to Norfloxacin (12.5). There were significant differences between class I and the two other classes of integrons (p=0.036). There was a statistically significant difference between the presence of blaTEM gene in Pseudomonas aeruginosa with other coding genes for antibiotic resistance (p=0.029). Conclusion: Pseudomonas aeruginosa isolates are multi-drug resistant and almost all isolates from clinical infections have class I, II and III Integrons

Detection of some virulence factors in Staphylococcus aureus isolated from clinical and subclinical bovine mastitis in Iran

Mastitis is one of the common diseases of dairy cattle and an inflammatory response of the mammary glands tissue. Mastitis causes considerable loss to the dairy industry. Among several bacterialpathogens that can cause mastitis, Staphylococcus aureus is probably the most lethal agent because it causes chronic and deep infection in the mammary glands that is extremely difficult to be cured. The present study was to detect some of the virulence factors in the S. aureus isolated from 360 mastitis milk samples in Chaharmahel va Bakhtiari and Isfahan provinces of Iran via PCR by using specific primers. Among a 360 raw milk samples, 86 samples contained 1250 bp fragment of the 23srRNA gene,42 samples contained coa gene, 63 samples contained clfA gene, 69 samples contained IgG binding region gene, 22 samples contained X region coding gene protein A, 3 sample contained Toxic shock syndrome toxin gene (tst), 16 samples contained the exfoliative toxin A and B genes, 10 samples contained agrI gene, 42 samples contained agrII gene, 19 samples contained agrIII gene and 15 samples contained agrIV gene

Effects of essential oils of Satureja bachtiarica and Nigella sativa on the efficacy of lactococcosis vaccine in rainbow trout (Oncorhynchus mykiss)

Lactococcosis has been defined as acute septicaemia, which causes economic losses in farmed fish, especially in rainbow trout. This study was done to evaluate the effects of the essential oils of Satureja bachtiarica and Nigella sativa on the efficacy of lactococcosis vaccine in rainbow trout. A total number of 270 fishes with a mean weight of 120 g were obtained; they were randomly divided into nine groups, each with three replicates, after two weeks of adaptation. The groups were: no injection group, vaccine only group, DMSO injection group, vaccine with 50, 100, and 200 micrograms Intraperitoneal injection (IP) injection. Two, four, and six weeks after vaccination, serological and haematological parameters were evaluated. In the sixth week, 1.7×10^7 cfu as LD50 96 hrs of Lactococcus garvieae were IP injected and the relative survival percentage was calculated. The results indicated that N. sativa essence is effective on the leukocyte population as the highest number of leukocytes were found in fish receiving high concentration of N. sativa. The relative survival rate of the studied fish decreased with decreasing concentrations of the N. sativa essential oil concentration, with a significant difference with control groups (p<0.05). However, using S. bachtiarica was not significantly effective on the relative survival rate of fish. The results of this study indicated that N. sativa essential oil can be used as adjuvant for L. garvieae vaccine, since it resulted in increasing leukocytes and the relative survival rate although S. bachtiarica was not effective on immune parameters of the studied fish

Conducting longitudinal research with older widows : Exploring personal communities through multiple methods

This article reports on the process of undertaking a longitudinal multiple methods study with older women experiencing the transition of later life widowhood. A series of three qualitative in depth interviews were conducted with twenty-six older widows in North Staffordshire, United Kingdom. Interviews included the use of personal community diagrams to identify the structure of personal communities, and Christmas and Christmas cards to further explore social relationships and practices during transition. Examples of cases are given to illustrate the findings derived from the methods employed. The cases demonstrate the diverse and often paradoxical nature of social relationships within similar networks

Human MLH1 Protein Participates in Genomic Damage Checkpoint Signaling in Response to DNA Interstrand Crosslinks, while MSH2 Functions in DNA Repair

DNA interstrand crosslinks (ICLs) are among the most toxic types of damage to a cell. For this reason, many ICL-inducing agents are effective therapeutic agents. For example, cisplatin and nitrogen mustards are used for treating cancer and psoralen plus UVA (PUVA) is useful for treating psoriasis. However, repair mechanisms for ICLs in the human genome are not clearly defined. Previously, we have shown that MSH2, the common subunit of the human MutSα and MutSβ mismatch recognition complexes, plays a role in the error-free repair of psoralen ICLs. We hypothesized that MLH1, the common subunit of human MutL complexes, is also involved in the cellular response to psoralen ICLs. Surprisingly, we instead found that MLH1-deficient human cells are more resistant to psoralen ICLs, in contrast to the sensitivity to these lesions displayed by MSH2-deficient cells. Apoptosis was not as efficiently induced by psoralen ICLs in MLH1-deficient cells as in MLH1-proficient cells as determined by caspase-3/7 activity and binding of annexin V. Strikingly, CHK2 phosphorylation was undetectable in MLH1-deficient cells, and phosphorylation of CHK1 was reduced after PUVA treatment, indicating that MLH1 is involved in signaling psoralen ICL-induced checkpoint activation. Psoralen ICLs can result in mutations near the crosslinked sites; however, MLH1 function was not required for the mutagenic repair of these lesions, and so its signaling function appears to have a role in maintaining genomic stability following exposure to ICL-induced DNA damage. Distinguishing the genetic status of MMR-deficient tumors as MSH2-deficient or MLH1-deficient is thus potentially important in predicting the efficacy of treatment with psoralen and perhaps with other ICL-inducing agents