Neuroprotective Effects of Heat Shock Protein70.

BACKGROUND & OBJECTIVE: Heat Shock Proteins (HSPs) increase response to many stresses in cells. Stroke is a neural shock that leads to the destruction of a large number of brain cells, whereas induction and expression of HSPs can decrease the amount of damage, and in some conditions can cure damaged cells. HSP70 family is considered as the most important member of HSPs in normal and stress condition of cells. They are strongly up-regulated by stresses and have protective roles in under stressed cells. Therefore, in this review, we briefly consider the association between HSP70 and stroke. We searched in Pubmed and Scopus databases using the specified keywords and selected the articles based on the certain association between HSP70 and stroke. HSP70 protects cells from damage through a variety of cellular and biochemical processes such as chaperone function, anti-apoptotic, anti-necrotic and anti-inflammatory mechanisms. CONCLUSION: Protective effects of HSP70 in neurodegenerative shocks are illustrated in the review, and it can be concluded that the induction of HSP70 in stresses can be considered as a therapeutic factor, although it needs further studies.

Effect of Fibrin Packing on Managing Hepatic Hemorrhage and Liver Wound Healing in a Model of Liver Stab Wound in Rat.

OBJECTIVE: To evaluate the effect of fibrin perihepatic packing on controlling liver hemorrhage and liver wound healing. METHODS: In this animal experimental study, 20 adult male Sprague Dawley rats, weighing 200-220 g, were included. Stab wound injury was created by number 15 scalpel, so that bilateral liver capsules and liver tissue were cut, and acute bleeding was accrued. The animals were divided into 2 study groups: control (with a primary gauze packing treatment) and test group (with fibrin packing treatment). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total serum bilirubin (TSB) levels were measured as a liver function test during the treatment period. Blood loss was calculated for estimation of hepatic hemorrhage during surgery. After four weeks, the liver wound repair was evaluated by sampling and Hematoxylin and Eosin staining (H&E). RESULTS: In the test group, all of animals were alive (mortality rate= 0%). Significantly, ALT and AST levels were raised after surgery, followed by a decrease ALT (p=0.783) and AST (p=0.947) to the normal level during 4 days. Estimated blood loss was 2.89 ± 0.73 mL (about 19.65% of estimated blood volume). Hematocrit levels returned to the normal level (p=0.109) after 48 hours. In the control group, the mortality rate was 50% during 12h after surgery. ALT (p=0.773) and AST (p=0.853) were decreased to normal level during 6 days, and estimated blood loss was 4.98 ± 0.77 mL (about 32.98% of estimated blood volume) in the remaining animals. Moreover, hematocrit levels returned to the normal level (p=0.432) after 72 hours. Estimated blood loss in the test group was significantly less than control group (p<0.001). Total serum bilirubin levels were not significantly different from the normal level, before and after surgery in both groups. Histopathology sections from the post-hepatectomy specimens showed that the site of the previous incision was completely repaired, and a dense fibrous septum was observed in both groups. CONCLUSION: The fibrin dressing was effective in preventing blood loss and saving lives after a liver stab injury and major internal bleeding in the animal model of rat.

Optimization of Three Dimensional Culturing of the HepG2 Cell Line in Fibrin Scaffold.

BACKGROUND: A potential treatment for healing hepatic tissue is delivering isolated hepatic cells to the site of injury to promote hepatic cells formation. In this technology, providing an appropriate injectable system for delivery of hepatic cells is an important issue. In this regard, fibrin scaffolds were designed with many advantages over other scaffolds like cell delivery vehicles for biodegradation, biocompatibility and hemostasis. OBJECTIVES: The aim of this study was to determine suitable cell culture circumstances for HepG2 cell proliferation and differentiation in 3D fibrin scaffolds by evaluating Ca(2+) concentrations, cell numbers, various ratios of plasma/RPMI 1640 and thickness of fibrin scaffold. MATERIALS AND METHODS: In a one-stage experimental design, Box-Behnken design strategy was performed by Minitab 15 software (version 15, Minitab. State College, PA) with three factors at three levels (low, medium and high) and 27 runs for identification of the effects of ratio of plasma/RPMI 1640, Ca(2+) concentration and thickness on the formation of fibrin gel scaffold and 3D HepG2 culture. RESULTS: The optimal concentrations for fibrin scaffold fabrication were achieved by adding 0.15 mol CaCl2 (50 µL) and 1 × 10(5) cells to 1:4 of plasma/RPMI 1640 ratio (500 µL with 2.3 mm thickness per well). CONCLUSIONS: Our approach provided easy handle method using inexpensive materials like human plasma instead of purified fibrinogen to fabricate fibrin scaffold.

Production of a novel multi-epitope peptide vaccine for cancer immunotherapy in TC-1 tumor-bearing mice.

In our previous research, several bioinformatic strategies were utilized to design an efficient multi-epitope peptide vaccine (MEV) against cancer. The designed vaccine consists of Wilms tumor-1 (WT-1) and human papillomavirus (HPV) E7 cytotoxic T lymphocyte (CTL) epitopes, tetanus toxin fragment C (TTFrC) and HLA-DR epitope (PADRE) helper T lymphocyte (HTL) epitopes and heparin-binding hemagglutinin (HBHA) as an immunostimulatory adjuvant. All segments were fused together by suitable linkers. In the current study, we cloned and expressed the designed MEV in E. coli. We subsequently performed in vivo preventative and therapeutic assays to evaluate antitumor efficacy of the vaccine against the HPV-16 E7-expressing murine tumor cell line TC-1 as a model for cancer immunotherapy. The results showed that in preventive experiments, vaccination with MEV significantly augmented the IgG antibody titer and the percentage of tumor-free mice compared to control groups (PBS and E7). Moreover, in therapeutic experiments, vaccination with MEV led to a reduction in the number of metastatic nodules, lung weights and the ratio of lung weights to body weights compared to other groups. In sum, our epitope vaccine could efficiently induce preventive and therapeutic antitumor immunity in TC-1 tumor bearing mice.

Association between tacrolimus concentration and genetic polymorphisms of CYP3A5 and ABCB1 during the early stage after liver transplant in an Iranian population.

OBJECTIVES: Tacrolimus is widely used as an immunosuppressive drug in liver transplant recipients with a narrow therapeutic range and variable individualized pharmacokinetics. Tacrolimus is a substrate of cytochrome P-450 3A enzyme and the drug transporter, P-glycoprotein. MATERIALS AND METHODS: We determined the genotypic frequencies of cytochrome P-4503A5 (rs776746), and ABCB1 (rs1045642), single nucleotide polymorphisms in a population of 100 Iranian liver transplant patients, and investigated the influence of the above-mentioned single nucleotide polymorphisms on tacrolimus concentrations. At 7 and 30 days after transplant, tacrolimus dosages (mg/kg/d), trough blood levels (T0), and dose-adjusted concentrations (concentration/dosage ratio) were determined. Polymerase chain reaction, followed by restriction fragment length polymorphism analysis, was used for genotyping cytochrome P-4503A5*3 [6986A>G] as well as ABCB1 [3435C>T]. RESULTS: Ninety-five percent of the population showed a cytochrome P-4503A5*3/*3 genotype. ABCB13435TT genotype was observed in 33 cases (33%); whereas 51 cases (51%) carried 3435CT, and 16 cases (16%) carried 3435CC. With regard to the ABCB1 and cytochrome P-4503A5, they showed no influence on tacrolimus dosing requirements at 1 week or 1 month after transplant. No association of any genetic variant with the acute rejection rate was found. CONCLUSIONS: Finally, as the liver donor genotype influences tacrolimus pharmacokinetics with regard to expression of cytochrome P-4503A5, far more than the genotype of the recipient; therefore, it should be considered before recommending any personal immunosuppressive treatment based on pharmacogenetics.

Human herpesvirus-6 viral load and antibody titer in serum samples of patients with multiple sclerosis.

BACKGROUND: Despite the number of cases with definite diagnosis of multiple sclerosis (MS) being on increase, the role of human herpesvirus-6 (HHV-6) infection as a trigger for MS disease still is deliberated. Based on antibody detection and quantitative HHV-6 polymerase chain reaction assay, this study was achieved to find out the possible association between infection with HHV-6 and clinical progression of MS disease. METHODS: A total of 108 serum samples were obtained from 30 MS patients followed prospectively for a 6-month period. These samples were analyzed for the presence of HHV-6 DNA by nested polymerase chain reaction enzyme-linked immunosorbent assay and for anti-HHV-6 IgG titer. Activation of the disease was determined by either magnetic resonance imaging or by clinical status of the patients. Control groups were also included. RESULTS: The average antibody index for the MS patients in the first sample collection was higher than both control groups (p = 0.001). HHV-6 DNA was detected in the serum samples of 10 of 30 MS patients. The mean HHV-6 viral load in patients with relapsing-remitting multiple sclerosis (RRMS) with and without relapse was 973 and 714, respectively. Seven patients showed an exacerbation during the study period. Of those, four patients had HHV-6 DNA in their collected samples. The prevalence of HHV-6 DNA was significantly higher in patients with MS as compared with control groups (p = 0.001). CONCLUSIONS: The results indicate that HHV-6 is implicated somehow in MS disease. Over time, rising HHV-6 IgG antibody titers together with an exacerbation and detection of HHV-6 DNA in serum samples of some MS patients suggests possible association between the reactivation of the virus and disease progression.

Study of viral infections in infants with biliary atresia.

Viral infections may have an important role in the pathogenesis of biliary atresia, and related clinical outcomes. In this research for determination of the possible role of HBV, HCV, HCMV, adenovirus, and BK virus infections in biliary atresia related clinical complications, the molecular and antigenic prevalence of these viral agents were studied. In this retrospective study, 34 formalin fixed paraffin embedded (FFPE) biopsy and autopsy liver tissue samples of neonates with biliary atresia were evaluated. The molecular prevalence of these viral infections was assayed by different PCR and RT-PCR methods. The antigenic prevalence of HBV, HCV, and HCMV infections was also studied in these liver tissue samples by immunohistochemistry (IHC) method. HBV, HCV, and adenovirus genomes were detected in 9%, 6%, and 6% of liver autopsy and biopsy tissues of infants with biliary atresia, respectively. HBV and HCV co-infection was confirmed in 6% of FFPE samples. The genome of other investigated viruses was not detected in FFPE liver tissues. Detection of viral infection in FFPE liver tissue samples of newborns with biliary atresia, suggests the need for complete studies for the determination of accurate role of these viral infections in pathogenesis of biliary atresia.

Value of serum cystatin C as a marker of renal function in the early post kidney transplant period.

Management of renal transplant patients requires periodic measurement of renal function especially in early post transplant period. This is usually assessed by measuring the creatinine clearance, but because of its limitations, it is not an ideal marker for assessing the renal function. Serum cystatin C (sCyC) appears to be an endogenous marker of glomerular filtration rate (GFR). To assess the use of sCyC as a marker of renal function in kidney transplant patients, we compared it with serum creatinine (sCr) and 24-hour urine creatinine clearance (CrCl) in the first week post-transplantation. Among 60 patients (62.8% men, 37.2% women) undergoing kidney transplantation (average age: 44.87 +/- 13.37 years), we determined renal function at 1, 3, 5, and 7 days after kidney transplantation using: sCr, sCyC and CrCl in a 24-hours urine specimen. During the first 5 days following transplantation, there was a progressive decline in sCr levels. In the first 5 days, post transplantation we could not find good correlation between CrC and sCyC, and the sCyC increased during these 5 days, but after that in day 7, there was a good correlation between CrC and sCyC which is coinciding with decreasing the dose of steroid (r= .625). Therefore, we recommend using sCyC may be used as a marker of renal function after one-week post kidney transplantation.