Benefits of conditioned medium of nicotine-pulsed mesenchymal stem cells in experimental autoimmune hepatitis.

Previous data indicated that nicotine could modulate the immune regulatory potential of mesenchymal stem cells (MSCs). Currently, we intend to assess the effects of a conditioned medium of nicotine-pulsed mesenchymal stem cells in the experimental model of autoimmune hepatitis (AIH). Bone marrow-derived MSCs pulsed with 0,.1,.5, or 1 µM nicotine until the cells reached 90% confluency. Correspondent to in vitro results, the least effective concentration of nicotine that led to an anti-inflammatory environment by the MSC-conditioned medium was 0.5 µM. The murine model of AIH induced by Intravenous injection Concanavalin A (ConA). Mice were allocated to pretreatment (Concomitant treatment with ConA administration) or treatment groups and received un-pulsed MSC-conditioned medium (CM) or conditioned medium of nicotine (0.5 µM)-pulsed MSCs (CMN). The levels of ALT, AST, MPO, TNF-α, IFN-γ, and IL-6 were the highest in the ConA group than in the other groups. Pretreatment or treatment with the CMN caused a significant reduction in hepatic enzymes and inflammatory cytokines compared to pretreatment or treatment with CM. Both CM or CMN significantly decreased the numbers of activated TCD4+ and TCD8+ in the blood. More importantly, pre-treatment or treatment with CMN caused a better improvement in the histopathological appearance than pre-treatment or treatment with CM. The results of this study show that CMN rapidly controls the AIH mouse model, and therefore it may be considered as a new therapeutic approach for the treatment of AIH patients.

Combined atorvastatin and pentoxifylline in ameliorating inflammation induced by complete Freund's adjuvant.

Injection of complete Freund's adjuvant (CFA) into one of the footpads of Wistar rats promotes swelling in all the footpads (including non-injected footpads) in the next few days, indicating a complicated immunological reaction and autoimmune arthritis. This survey was performed to assess the synergistic benefits of combined atorvastatin and pentoxifylline for treating arthritis induced by CFA. Therapeutic regimes began on day 12 post-challenge when all the rats recorded an arthritis index of more than one and continued throughout the investigation until day 32. Treatment with combined atorvastatin and pentoxifylline at half doses led to synergistic benefits causing the regression of clinical and radiological appearance of arthritis in non-injected paws, which was more favorable than treatment with any medication alone at full doses. Moreover, the combined treatment led to a reduction in some inflammatory biochemical parameters, such as myeloperoxidase, nitric oxide, and C-reactive protein, which was more pronounced than those of the treatment with each medication alone. The mRNA expression of IL-1ß and TNF-α in the rat toe area was significantly decreased by combination therapy, and this reduction was more significant than that of monotherapy. The ratios of RORγc/T-bet, RORγc/GATA-3, RoRγc/Foxp3, T-bet/GATA-3, and T-bet/Foxp3 expression showed a further decrease in the combined treated group compared to other groups. Interestingly, combination therapy did not reduce T lymphocyte proliferation to a level lower than healthy rats. Overall, the combination of atorvastatin and pentoxifylline possesses immunomodulatory synergistic benefits, and this approach may be an impressive strategy to manage complicated inflammation.

Boosting effect of testosterone on GDNF expression in Sertoli cell line (TM4); comparison between TM3 cells-produced and exogenous testosterone.

The Glial cell-derived neurotrophic factor (Gdnf) and testosterone induce the spermatogonial stem cells (SSCs) self-renewal and spermatogenesis, respectively. In present study the stimulating role of testosterone on Sertoli cells to produce Gdnf, and the possible effect of Gdnf on Gfrα1 and c-RET expressions were investigated. The TM4 cells (line Sertoli cells) were co-cultured with [0.1, 0.2 and 0.4 (ng/ml)] of exogenous and TM3 (line Leydig cells)-produced testosterones, and consequently the TM4-produced Gdnf concentration was evaluated. Next, the SSCs were co-cultured with the TM-4 derived media (endogenous Gdnf) and exogenous Gdnf [0.1, 0.2, and 0.4 ng/ml)]. The 0.1 and 0.2 ng/ml endogenous and 3 concentrations of exogenous testosterone up-regulated the Gdnf expression versus non-treated Sertoli cells. The TM4-produced and exogenous Gdnfs, in all concentrations, up-regulated the receptors expression. In conclusion, the testosterone, solely, stimulates the Gdnf synthesis and the Gdnf, individually, amplifies its receptor's expression at mRNA and protein levels.

Mutual relationship between surface atmospheric pollutants and CG lightning in Tehran area.

The mutual relationship between surface atmospheric pollutants and Cloud-to-Ground (CG) lightning is studied in the Tehran area for the first time. The impact of surface pollutant concentration of PM10 on CG lightning initiation, as well as the production of NO2 and surface ozone derived by lightning occurrence is investigated. To proceed, the reports of thunderstorm occurrence, including CG lightning in the Tehran area have been collected for years of study from the Iran meteorological organization (IRIMO). The surface pollution data are obtained from the Air Quality Control Center (AQCC) for several stations in the area of interest. The number of lightning (NoL) associated with the selected dates is obtained using the World Wide Lightning Location Network (WWLLN). The correlation coefficients associated with the CG lightning and the concentrations of PM10, NO2, and surface ozone are calculated. The hourly variations of accumulated NoL and NO2 and surface ozone are also compared for 24 hours before and after the lightning activity for four days of study during the years of 2009-2013. The results show that there is a positive correlation between PM10 concentration and lightning flash number, obtained from observational data of WWLLN. Moreover, the comparison of NoL and surface pollutant concentration indicates a clear positive contribution from CG lightning in NO2 and ozone production. In days with a considerable number of lightning occurrences, the comparison of the hourly average of NO2 and O3 concentrations with the lightning flash number reveals that NO2 decreases, and O3 increases due to the significant increases of lightning strikes.

Morphologic and molecular analysis of Dicrocoelium dendriticum species from Iranian native sheep.

Dicrocoeliosis is a worldwide parasitic disease of ruminants which affects the liver. In this current study, the phylogenic pattern of Dicrocoelium species in Iranian native sheep from three different geographical regions was investigated by investigating a 520 bp fragment of mitochondirial NAD1 gene. The analysis of the NAD1 oligo nucleotide sequences from 10 D. dendriticum (GenBank accession numbers: MG889399 to MG889408) revealed few non-significant differences, suggesting limited application for NAD1 gene as a biomarker for study of genetic variation in Dicrocoelium. The morphometrical study also showed a significant relationship for the several morphometric indices among the Dicrocoelium spp. isolates from different regions of Iran.

Testosterone amplifies HSP70-2a, HSP90 and PCNA expression in experimental varicocele condition: Implication for DNA fragmentation.

The DNA fragmentation and failure in post-meiotic maturation of the spermatozoa because of testosterone withdrawal can affect the fertilization potential in varicocele (VCL) patients. To find out the exact mechanism of VCL-induced failure in histone-protamine replacement process and DNA fragmentation, the correlations between the levels of expression of HSP70-2a, HSP90, PCNA, TP1/2 and PCNA genes and the patterns of DNA methylation were investigated before and after testosterone administration in rats. In total, 40 mature male Wistar rats (10 in each group) were assigned between control (with no intervention), control-sham (undergone a simple laparotomy), VCL-induced (VCL-sole), and testosterone-treated VCL-induced (VCLT) groups. The HSP70-2a, HSP90, PCNA, TP1, and TP2 genes expressions and the patterns of global DNA methylation were determined in all groups. A statistically significant (p < 0.05) reduction were found in the HSP70-2a, HSP90, PCNA, TP1 and TP2 genes expressions in VCL-sole group. In VCLT group, testosterone was shown to significantly (p < 0.05) up-regulate the HSP70-2a, HSP90, PCNA, and TP2expression levels, but TP1 expression has not been changed. Furthermore, the VCLT group exhibited higher DNA methylation rates compared to VCL-sole animals. In conclusion, testosterone, by up-regulating the HSP70-2a and HSP90 expressions and maintaining the pre-existing HSP70-2a and HSP90 proteins levels, may be the reason for the significant increment in TP2 expression during post-meiotic stage and can boost the global methylation rates of DNA via up-regulating the PCNA expression, suggesting that administration of testosterone can mitigate the VCL-impaired histone-protamine replacement and DNA methylation rates and protect the cellular DNA content from VCL-induced oxidative stress.

Chitosan-starch film containing pomegranate peel extract and Thymus kotschyanus essential oil can prolong the shelf life of beef.

The aim of the current study was to investigate the effects of pomegranate peel extract (PPE) and chitosan-starch (CH-S) composite film incorporated with Thymus kotschyanus essential oil (TEO) on the shelf-life of beef during storage period of 21 days at 4 °C. The physico-mechanical parameters, the color and Fourier Transform Infrared spectra values of the films were determined. Changes in odor, color, pH, thiobarbituric value for lipid oxidation levels and Pseudomonas spp. total viable counts, lactic acid and Listeria monocytogenes were determined during the storage time. All treated films showed lower elongation, strength and transparency values compared with chitosan - starch (CH-S) group. Results showed that CH-S-PPE 1%-TEO 2% treatment had the highest inhibition effect against L. monocytogenes. The bacterial counts and lipid oxidation were successfully inhibited using PPE and TEO. CH-S-PPE 1% group containing up to 2% TEO had the best acceptable sensory characteristic. Beef samples wrapped with CH-S film containing PPE and TEO also had longer shelf life.

Differential transcription patterns in wild-type and glycoprotein G-deleted infectious laryngotracheitis viruses.

Infectious laryngotracheitis virus (ILTV) causes severe respiratory disease in poultry throughout the world. Recently the role of glycoprotein G (gG) in ILTV pathogenesis has been investigated and it has been shown to have chemokine-binding activity. An ILTV vaccine candidate deficient in gG has been developed and the deletion has been shown to alter the host's immune response to the virus. To understand the effect of the gG gene on transcription of other viral genes, the global expression profile of 72 ILTV genes in gG-deleted and wild-type ILTVs were investigated both in vivo and in vitro using quantitative reverse transcription-polymerase chain reaction. Several genes were differentially expressed in the different viruses in LMH cell cultures or in the tracheas of infected birds, and the expression of a number of genes, including ICP27, gC, gJ, Ul7 and UL40, differed significantly both in vivo and in vitro, suggesting that they had direct or indirect roles in virulence. This study has provided insights into the interactions between gG and other ILTV genes that may have a role in virulence.

Association between Anemia and COPD in Iranian Population.

BACKGROUND AND AIM: Chronic obstructive pulmonary disease (COPD) is one of the major causes of morbidity and mortality in adults. Anemia is known as comorbidity in many chronic diseases that can increase morbidity and mortality of COPD. Recent studies have shown that anemia may be more prevalent than expected in COPD patients and can increase disabilities of COPD. In this study we have evaluated the correlation between anemia and the severity of COPD in patients referred to teaching hospitals of the Tehran University of Medical Sciences (TUMS), Tehran, Iran. MATERIALS AND METHODS: In this cross-sectional study the severity of COPD in 760 patients with dyspnea who referred to teaching hospitals of Tehran University of Medical Sciences and 96 stable COPD patients were categorize using a GOLD criteria from mild to moderate, severe and very severe. Anemia was determined as hemoglobin <13 g/dL in men and <12 g/dL in women, respectively. Demographic characteristics, spirometry parameters and laboratory findings were compared between anemic and non-anemic groups using Student t-test and regression tests (SPSS v.18 software). RESULTS: The Mean age of patients was 65 ± 13.07 years (59.4% male). Overall prevalence of anemia was 27% and there was no correlation between severity of COPD and anemia. Anemic patients were significantly older than non-anemic patients (71.1 ± 8.5 years vs. 65.4± 12.8 years; p = 0.030). RBC count of anemic patients were significantly lower than non-anemic group (4.3 ± 0.5 vs. 5.02± 0.8 ×106/µL; p < 0.001). Erythropoietin levels in anemic group was significantly higher than non-anemic group (16.33±2.43 vs. 10.22 ± 2.67 mu/ml; p < 0.001) and there was a significant inverse correlation of hemoglobin vs erythropoietin (r= -0.8). CONCLUSION: There was a high prevalence of anemia in COPD patients. Anemia can increase disabilities of COPD. Thus, treatment of anemia may improve quality of life in these patients. Further comprehensive studies are needed for determination of exact prevalence of anemia and its physiologic effects in COPD.

Prevalence of peripheral arterial disease in hemodialysis patients.

INTRODUCTION: The ankle-brachial index (ABI), measurement of carotid artery intima-media thickness (CIMT), and assessment of the thickness of interventricular septum (IVS), are noninvasive methods used to predict subclinical atherosclerosis in hemodialysis patients. This study aimed to determine the prevalence of peripheral arterial disease and to assess the correlations between ABI, CIMT, the thickness of IVS, and blood parameters in hemodialysis patients. MATERIALS AND METHODS: The ABI, CIMT, and the thickness of IVS were measured in 50 patients on hemodialysis. Data were collected regarding the levels of calcium, urine nitrogen, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglyceride, cholesterol, creatinine, albumin in serum, as well as erythrocyte sedimentation rate. Results. Ten percent of the patients showed a reduced ABI (< 0.9). The mean values for ABI, CIMT, and IVS were 1.09 ± 0.13, 0.68 ± 0.11 mm, and 9.83 ± 1.65 mm, respectively. The levels of calcium, cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglyceride in the serum of the patients with normal ABI were significantly higher than in patients with reduced ABI. There was a negative correlation between ABI and levels of serum LDLC (r = -0.29, P = .04) and triglyceride (r = -0.32, P = .02). Conclusions. The prevalence of peripheral arterial disease in the patients with CRF was 10% and it was correlated with several classical risk factors for atherosclerosis, including elevated LDL and cholesterol levels. CIMT and the thickness of IVS showed no apparent association with ABI.

Effects of valerian consumption during pregnancy on cortical volume and the levels of zinc and copper in the brain tissue of mouse fetus.

OBJECTIVE: The aim of the present study was to determine the effects of valerian (Valeriana officinalis) consumption in pregnancy on cortical volume and the levels of zinc and copper, two essential elements that affect brain development and function, in the brain tissues of mouse fetuses. METHODS: Pregnant female mice were treated with either saline or 1.2 g/kg body weight valerian extract intraperitoneally daily on gestation days (GD) 7 to 17. On GD 20, mice were sacrificed and their fetuses were collected. Fetal brains were dissected, weighed and processed for histological analysis. The volume of cerebral cortex was estimated by the Cavalieri principle. The levels of zinc and copper in the brain tissues were measured by atomic absorption spectroscopy. RESULTS: The results indicated that valerian consumption in pregnancy had no significant effect on brain weight, cerebral cortex volume and copper level in fetal brain. However,it significantly decreased the level of zinc in the brain (P<0.05). CONCLUSION: Using valerian during midgestation do not have an adverse effect on cerebral cortex; however,it caused a significant decrease in zinc level in the fetal brain. This suggests that valerian use should be limited during pregnancy.

Kinetics of transcription of infectious laryngotracheitis virus genes.

The kinetics of expression of only a few genes of infectious laryngotracheitis virus (ILTV) have been determined, using northern blot analysis. We used quantitative reverse transcriptase PCR to examine the kinetics of expression of 74 ILTV genes in LMH cells. ICP4 was the only gene fully expressed in the presence of cycloheximide, and thus classified as immediate-early. The genes most highly expressed early in infection, and thus classified as early, included UL1 (gL), UL2, UL3, UL4, UL5, UL6, UL7, UL8, UL13, UL14, UL19, UL20, UL23 (TK), UL25, UL28, UL29, UL31, UL33, UL34, UL38, UL39, UL40, UL42, UL43, UL44 (gC), UL47, UL48 (α-TIF), UL49, UL54 (ICP27), US3 and US10. ORF A, ORF B, ORF C, ORF E, sORF 4/3, UL[-1], UL0, UL3.5, UL9, UL10 (gM), UL11, UL15a, UL15b, UL18, UL22 (gH), UL24, UL26, UL30, UL32, UL36, UL45, UL49.5 (gN), UL52, US2, US4 (gG), US5 (gJ) and US9 were most highly expressed late in infection and were thus considered late genes. Several genes, including ORF D, UL12, UL17, UL21, UL27 (gB), UL35, UL37, UL41, UL46, UL50, UL51, UL53 (gK), US8 (gE), US6 (gD) and US7 (gI), had features of both early and late genes and were classified as early/late. Our findings suggest transcription from most of ILTV genes is leaky or subject to more complex patterns of regulation than those classically described for herpesviruses. This is the first study examining global expression of ILTV genes and the data provide a basis for future investigations of the pathogenesis of infection with ILTV.

Development of a SYBR Green quantitative polymerase chain reaction assay for rapid detection and quantification of infectious laryngotracheitis virus.

Infectious laryngotracheitis is an acute viral respiratory disease of chickens with a worldwide distribution. Sensitive detection of the causative herpesvirus is particularly important because it can persist in the host at a very low copy number and be transmitted to other birds. Quantification of viral genome copy number is also useful for clinical investigations and experimental studies. In the study presented here, a quantitative polymerase chain reaction (qPCR) assay was developed using SYBR Green chemistry and the viral gene UL15a to detect and quantify infectious laryngotracheitis virus (ILTV) in ILTV-inoculated chicken embryos or naturally infected birds. The specificity of the assay was confirmed using a panel of viral and bacterial pathogens of poultry. The sensitivity of the assay was compared with two conventional PCR assays, virus titration and an antigen-detecting enzyme-linked immunosorbent assay. The qPCR developed in this study was highly sensitive and specific, and has potential for quantification of ILTV in tissues from naturally and experimentally infected birds and embryos.

Relationship between mortality, clinical signs and tracheal pathology in infectious laryngotracheitis.

Previous studies in our laboratory using a combination of polymerase chain reaction and restriction fragment length polymorphism have identified at least five different genotypes of infectious laryngotracheitis virus (ILTV). However, the virulence of these classes of ILTV was not investigated. In this study, five groups (16 birds each) of 3-week-old specific pathogen free chickens were inoculated via the intratracheal route with 10(3) median embryo infected dose of five different strains of ILTV. Three further groups of chickens were inoculated similarly with the vaccine strains SA2 and A20 or with sterile phosphate-buffered saline (PBS) for comparison. Four days post-inoculation, clinical signs were monitored for scoring, and eight chickens from each group were subsequently euthanized, weighed and subjected to pathological and histopathological examinations. The remaining birds were monitored for clinical signs and mortality until 21 days post-inoculation. All groups inoculated with ILTV strains showed moderate to severe clinical signs 4 days after inoculation. The strain Q1-96 caused only minimal breathing symptoms with a median score that was not significantly different to that of the group inoculated with PBS, but was significantly different to those of the groups inoculated with other ILTV strains. The strain Q1-96 caused severe photophobia and conjunctivitis with a median score that was significantly higher than those of all other groups except for the group inoculated with the strain N3-04. All ILTV strains caused a significant reduction in weight gain when compared with the group inoculated with PBS. The strain Q1-96 caused an average weigh loss of 14% that was significantly higher than those of other ILTV strains. The strains S2-04 and Q1-96 induced only minor microscopic tracheal lesions while all the other ILTV strains, including the vaccine strains A20 and SA2, induced moderate to severe microscopic tracheal lesions. Median scores for microscopic tracheal lesions were well correlated with the number of viral genomes detected in trachea. The results revealed that there is considerable variation among ILTV strains in their tropism for trachea or conjunctiva. In addition it was revealed that ILTV strains with high affinity for conjunctiva can severely affect weigh gain. The ILTV numbers and microscopic lesions in trachea were not found to be reliable indicators of virulence since they are not necessarily correlated with mortality rate in ILT.

Differentiation of infectious laryngotracheitis virus isolates by restriction fragment length polymorphic analysis of polymerase chain reaction products amplified from multiple genes.

Infectious laryngotracheitis (ILT) has been identified in most countries around the world and remains a threat to the intensive poultry industry. Outbreaks of mild to moderate forms of ILT are common in commercial layer flocks, while sporadic outbreaks of ILT in broiler flocks have also been recognized as an emerging problem in several countries. Examination of viral isolates using restriction fragment length polymorphism of polymerase chain reaction (PCR-RFLP) from individual ILTV genes has suggested that some of these outbreaks were caused by vaccine strains. In this study, PCR-RFLP of a number of ILTV genes/genomic regions including gE, gG, TK, ICP4, ICP18.5, and open reading frame (ORF) B-TK was used to examine a number of historical and contemporary Australian ILTV isolates and vaccine strains. PCR-RFLP of gE using restriction endonuclease EaeI failed to distinguish between any of the isolates including the vaccine strains. PCR-RFLP of gG, TK, and ORFB-TK using restriction endonucleases MspI and FokI, respectively, divided all the isolates into two groups. PCR-RFLP of ICP18.5 and ICP4 using restriction endonuclease HaeIII separated the isolates into three different groups with some field isolates only able to be distinguished from vaccine strains by PCR-RFLP of ICP18.5. A combination of groupings including gG, TK, ICP4, ICP18.5, and ORFB-TK PCR-RFLP classified the ILTV isolates under investigation into five different groups with most isolates distinguishable from vaccine strains. Results from this study reveal that to achieve reliable identification of strains of ILTV, the examination of multiple gene regions will be required, and that most of the recent ILT outbreaks in Australia are not being caused by vaccine strains.