Comparative expression analysis of HSP7o, HSP9o, IL-4, TNF, A777<7 and KIT-receptor G and n and between varicocele-induced and non-varicocele testes of dog

Background: This study was designed to create an experimental varicocele model by a simple surgical procedure in dog with minimum invasion and to investigate the effect of varicocele-induced infertility on the expression of six related genes [HSP90, HSP70, IL-4, TNF, KITLG and KIT receptor]

Materials and Methods: In this experimental study, the proximal part of the pampini-form plexus of dog testes was partially occluded without abdominal incision which was confirmed by venographic examination. To evaluate varicocele in its acute form, dogs were castrated after 15 days and testes were dissected. Histopathologic evaluation was undertaken and the relative expression of the six genes was assessed by quantitative real-time polymerase chain reaction [PCR]

Results: Microscopic changes showed tubule degeneration. The Johnson score was significantly decreased in the varicocele testes when compared with non-varicocele testes. Expressions of HSP90, TNF, KITLG and the KIT-receptor gene were significantly down-regulated [P=0.029,0.047,0.004 and 0.035 respectively] in varicocele-induced testes while HSP70 was upregulated [P=0.018]. IL-4 did not show differential expression [P=0.377]

Conclusion: We conclude that partial occlusion of the proximal part of the pampiniform plexus induces varicocele in the testis of dog. Differential expression of the mentioned genes may be responsible for the pathophysiology of varicocele and related subfertility

Correlation of adiponectin RNA abundance and its receptors with quantitative parameters of sperm motility in rams

Background: Adiponectin and its receptors [AdipoRl and AdipoR2], known as adiponectin system, have some proven roles in the fat and glucose metabolisms. Several studies have shown that adiponectin can be considered as a candidate in linking metabolism to testicular function. In this regard, we evaluated the correlation between sperm mRNA abundance of adiponectin and its receptors, with sperm motility indices in the present study

Materials and Methods: In this completely randomized design study, semen samples from 6 adult rams were fractionated on a two layer discontinuous percoll gradient into high and low motile sperm cells, then quantitative parameters of sperm motility were determined by computer-assisted sperm analyzer [CASA]. The mRNA abundance levels of Adiponectin, AdipoRl and AdipoR2 were measured quantitatively using real-time reverse transcriptase polymerase chain reaction [qRT-PCR] in the high and low motile groups

Results: Firstly, we showed that adiponectin and its receptors [AdipoRl and AdipoR2] were transcriptionally expressed in the ram sperm cells. Using Pfaff based method qRT-PCR, these levels of transcription were significantly higher in the high motile rather than low motile samples. This increase was 3.5, 3.6 and 2.5 fold change rate for Adiponectin, AdipoRl ar/dAdipoR2, respectively. Some of sperm motility indices [curvilinear velocity [VCL], straight-line velocity [VSL], average path velocity [VAP], linearity [LIN], wobble [WOB] and straightness [STR]] were also significantly correlated with Adiponectin and AdipoRl relative expression. The correlation of AdipoR2 was also significant with the mentioned parameters, although this correlation was not comparable with adiponectin and AdipoRl

Conclusion: This study revealed the novel association of adiponectin system with sperm motility. The results of our study suggested that adiponectin is one of the possible factors which can be evaluated and studied in male infertility disorders

Comparison of proliferative and multilineage differentiation potential of sheep mesenchymal stem cells derived from bone marrow, liver, and adipose tissue

Despite major progress in our general knowledge related to the application of adult stem cells, finding alternative sources for bone marrow Mesenchymal Stem Cells [MSCs] has remained to be challenged. In this study successful isolation, multilineage differentiation, and proliferation potentials of sheep MSCs derived from bone marrow, adipose tissue, and liver were widely investigated. The primary cell cultures were prepared form tissue samples obtained from sheep 30-35 day fetus. Passage-3 cells were plated either at varying cell densities or different serum concentrations for a week. The Population Doubling Time [PDT], growth curves, and Colony Forming Unit [CFU] of MSCs was determined. The stemness and trilineage differentiation potential of MSCs were analyzed by using molecullar and cytochemical staining approaches. The data was analyzed through one way ANOVA using SigmaStat [ver. 2]. The highest PDT and lowest CFU were observed in adipose tissue group compared with other groups [p<0.001]. Comparing different serum concentrations [5, 10, 15, and 20%], irrespective of cell sources, the highest proliferation rate was achieved in the presence of 20% serum [p<0.001]. Additionally, there was an inverse relation between cell seeding density at culture initiation and proliferation rate, except for L-MSC at 300 cell seeding density. All three sources of fetal sheep MSCs had the identical trilineage differentiation potential. The proliferative capacity of liver and bone marrow derived MSCs were similar at different cell seeding densities except for the higher fold increase in B-MSCs at 2700 cells/cm2 density. Moreover, the adipose tissue derived MSCs had the lowest proliferative indices

Idiopathic retinal vasculitis, aneurysms and neuroretinitis syndrome associated with positive perinuclear antineutrophil cytoplasmic antibody

To report a case of idiopathic retinal vasculitis, aneurysms and neuroretinitis [IRVAN] syndrome associated with positive perinuclear antineutrophil cytoplasmic antibody [P-ANCA]. A 51-year-old man presented with loss of vision in his right eye since many years ago and blurred vision in his left eye over the past year. Ophthalmologic examination revealed optic atrophy and old vascular sheathing in the right eye and blurred disc margin, macular exudation, flame shaped hemorrhages, retinal vascular sheathing and multiple aneurysms at arterial bifurcation sites in the left eye, findings compatible with IRVAN syndrome. On systemic workup, the only notable finding was P-ANCA positivity. IRVAN syndrome may be a retinal component of P-ANCA associated vasculitis

[Semiquantitative RT-PCR analysis to assess the expression levels of iNOS in chicken macrophages]

iNOS is inducible by a variety of factors related to inflammation and referred to as inducible NOS[iNOS]. It is regulated at the level of gene expression; once expressed, it produces NO at a high rate. iNOS gene-expression profiling is an important tool in understanding molecular markers of the responses of cells and tissues to external factors. In this article a semiquantitative reverse transcription-polymerase chain reaction [RT-PCR] protocol was optimized to extract RNA [ribonucleic acid] from chicken spleen and to measure the expression levels of iNOS mRNAs from each sample. Detailed procedure was described for the analysis of iNOS levels. beta-actin was used as an internal control to normalize for sample to sample variations in total RNA amounts and for reaction efficiency. Co-amplification of the iNOS gene with housekeeping gene [beta-actin] provides a quantitative result. Changes in gene expression level may be monitored, while avoiding sample-to-sample loading variation