Comparison of Three Different Methods of Transfection for the Production of Recombinant Adenovirus Expressing Human Carcinoembryonic Antigen Gene.

Adenoviral vectors (AdVs) are widely used as a gene delivery vehicle and vaccine design due to their genetic stability, transfer capacity of large genes, production at high titers, and remarkable efficacy of transduction. One of the most important applications of AdVs is in cancer immunotherapy. Tumor-associated antigens are overexpressed in cancer cells; however, they cannot induce immune responses sufficiently. Therefore, the immune system must be stimulated against these antigens to kill the cancer cells. This study described the construction steps of a recombinant AdV expressing human carcinoembryonic antigen (CEA) gene. Furthermore, in order to achieve a high titer of the virus, an efficient transfection was required. Three various transfection reagents were compared to achieve the best method of transfection. Carcinoembryonic antigen was cloned into the pAdV and transfected into the A293 cells using three different reagents, including polyethylenimine (PEI), calcium phosphate, and DMRIE-C. The PEI had the highest transfection efficiency, which was selected for the transfection of the recombinant plasmid. It has low toxicity for cells and is suitable for large-scale transfection. The virus produced in this study can be applied as a vaccine in cancer immunotherapy for stimulating the immune system against CEA-expressing tumors.

Intra-arterial transplantation of neural stem cells improve functional recovery after transient ischemic stroke in adult rats.

OBJECTIVES: The neural stem cell transplantation has been proposed as alternative therapy to promote functional recovery after various neurological disorders. The aim of this study was to evaluate the effect of intra-arterial transplantation of adult neural stem cells on improving local brain ischemia injuries. MATERIALS AND METHODS: In this study, 32 male Wistar rats were used. Ischemia was induced using a middle cerebral artery obstruction with monofilament nylon suture. Neural stem cells were isolated from subventricular zone of the rat brain. 24 hours after local ischemia, the cells were labeled with DiI and transplanted intra-articularly. Evaluation of neurological movement deficits was performed using a neurological deficit score. The transplanted neural stem cells differentiation into neurons and astrocytes was assessed by immunohistochemistry. RESULTS: The results indicated that lesion volumes in the ischemic control, PBS and the treatment groups were 31.5, 29.8 and 14.7 % respectively. Our results also showed that the number of eosinophilic neurons and also neurological impairment in the treatment group was significantly reduced compared to the control and PBS groups. CONCLUSION: The results suggested that intra-arterial transplantation of neural stem cells 24 hours after ischemia, led to a decrease in the volume of brain ischemic lesion and improved neurological outcomes (Fig. 7, Ref. 23).

Observation period for asymptomatic penetrating chest trauma: 1 or 3 h?

BACKGROUND: Current recommendations for evaluation and safe discharge of penetrating chest trauma patients regarding pneumothorax (PTX) include a Chest X Ray (CXR) at the Emergency Department (ED) upon arrival and second CXR after 3 h if the first one is negative. PURPOSE: To compare CXRs taken at the first and third hours of ED arrival and evaluate a 1 h period of observation instead of 3 h for safe discharge of patients with penetrating chest trauma. METHODS: In this cross-sectional study, all asymptomatic patients with penetrating chest trauma referred to a level 1 trauma center with negative initial Postero-Anterior (PA) CXRs (hour 0) were enrolled. Those with intoxication, tube thoracostomy, chest computed tomography, evidence of abdominal penetration, an overall elapsed timed of more than 1 h for admission to the ED, and refusal to take part in the study were excluded. Patients underwent subsequent PA CXRs at hours 1 and 3. A phone call follow up after 24 h was organized for each patient. RESULTS: A total of 68 patients were enrolled. There was 100 % concordance among CXRs performed at hours 1 and 3 in the study population. None of the patients showed clinical deterioration or PTX in CXR at hour 1 if remained asymptomatic during the first hour of observation. CONCLUSION: Asymptomatic patients with penetrating chest trauma, negative initial PA CXR, no signs of intoxication, and no deterioration during the first hour of observation may be considered for discharge. Further evidence is required to make recommendations based on these findings.

Nortriptyline protects testes against germ cell apoptosis and oxidative stress induced by testicular ischaemia/reperfusion.

We designed this experiment to evaluate the effects of nortriptyline on testicular injury after torsion/detorsion (T/D). Ninety-six adult Wistar rats were divided into six groups 16 each in control group (Group 1), sham operated (Group 2), T/D + saline (Group 3), and in groups 4-6; were administered 2, 10 and 20 mg kg-1 , i.p. of nortriptyline 30 and 90 min after torsion respectively. Testicular torsion was created by twisting the right testis 720° in clockwise direction for 1 h. In six rats of each group, tissue MDA level and caspase-3 activity increased and the activities of catalase, superoxide dismutase and glutathione peroxidase decreased in compared with control group 4 h after detorsion (P < 0.001). In six rats of each group 24 h after detorsion, histopathological changes and germ cell apoptosis were significantly deteriorated by measuring mean of seminiferous tubules diameters (MSTD) and TUNEL test. Moreover, 30 days after T/D, sperm concentration and motility were examined in rest of rats. Pre- and post-reperfusion nortriptyline could reduce MDA and caspase-3 levels and normalise antioxidant enzymes activities, dose dependently. Germ cell apoptosis was significantly decreased, and the MSTD, as well as sperm functions, were significantly improved. Inhibition of mitochondrial permeability transition pore is probably involved in protective effects of nortriptyline against testicular T/D cell damages.

Efficacy of Crocus sativus L. on reduction of cadmium-induced toxicity on spermatogenesis in adult rats.

Cadmium is a toxic heavy metal element, which probably cause infertility by impairment in spermatogenesis. The present work aimed (i) to study the toxic effect of cadmium on spermatogenesis in rat, as well as (ii) the protective effect of Crocus sativus L. on cadmium-intoxicated rats. Cadmium chloride was administered intraperitoneally during 16 days at intervals of 48 h between subsequent treatments. Crocus sativus L. was pre-treated in both of control and cadmium-injected rats. Animals were sacrificed on day 17 after the first treatment. The left cauda epididymis was removed and immediately immersed into Hank's balanced salt solution for the evaluation of sperm count and viability, and left testis was fixed in 10% formalin for histological evaluation. Following contamination with cadmium, a decrease was observed in the number and viability of cauda epididymis sperm, which were increased by Crocus sativus L. pre-treatment (P < 0.05). In addition, cadmium decreased both cell proliferation and Johnsen Scores in the seminiferous tubules, which were reversed by Crocus sativus pre-treatment (P < 0.05). Furthermore, cadmium-induced decrease in the amount of free serum testosterone as well as an increase in lipid peroxidation activity in the testicular tissue was reversed by Crocus sativus L. (P < 0.05). These findings may support the concept that Crocus sativus L. can improve the cadmium toxicity on spermatogenesis.

Life cycle characteristics of six Artemia populations from Iran.

The present study was conducted to achieve the life cycle characteristics of six Artemia populations (one bisexual and five parthenogenetic) from Iran. The cysts of parthenogenetic strains were collected from Maharlu, Incheh, Varmal and Qom salt lakes and Lagoons at the periphery of Lake Urmia. Cysts of the bisexual Artemia urmiana were collected from the Lake Urmia. All cysts strains were hatched using the standard procedures and the nauplii from all populations were reared at 80 g L(-1) at laboratory conditions. Survival and total length of the Artemia were measured on days 8, 11, 14, 17, 20 and 23 of culture. Randomly selected adult animals were studied for eight reproductive and four life span characteristics. The findings showed that parthenogenetic Artemia from Maharlu, Incheh lakes and from Lagoons at the vicinity of the Lake Urmia had significantly highest (p < 0.05) values of survival rate (73 and 62.8%, respectively) compared to bisexual A. urmiana and parthenogenetic strains from Qom and Varmal lakes (49.6, 29.2 and 23.2%). No significant differences were observed in all growth strain populations when cultured under similar laboratory conditions. Artemia populations from Maharlu, Qom lakes and from Lagoons in many occasions had significantly highest (p < 0.05) reproductive values compared to other three populations including the bisexual A. urmiana. The results showed the highest heterogeneity and intrapopulation variations among parthenogenetic population strains.