comparison of the multiple oocyte maturation gene expression patterns between the newborn and adult mouse ovary

The interaction between follicular cells and oocyte leads to a change in gene expression involved in oocyte maturation processes. The purpose of this study was to quantify the expression of more common genes involved in follicular growth and oocyte developmental competence. In this experimental study, the expression of genes was evaluated with qRT-PCR assay in female BALB/c mice pups at 3-day of pre-pubertal and 8 week old virgin adult ovaries. The tissue was prepared by H and E staining for normal morphological appearance. The data were calculated with the 2-delta Ct formula and assessed using non-parametric two-tailed Mann-Whitney test. The p<0.05 was considered as significant. The data showed a significant increase in the level of Stra8 and GDF9 in adult compared with newborn mice ovaries [p=0.049]. In contrast, a significant decrease in the level of Mvh, REC8, SCP1, SCP3, and ZP2 was observed in adult mice ovaries compared to those in the newborn mice ovaries [all p=0.049 except SCP1: p=0.046]. There was no significant difference in the level of OCT4 and Cx37 expression between adult and newborn mice ovaries. The modifications in gene expression patterns coordinate the follicular developmental processes. Furthermore, the findings showed higher expression level of premeiotic gene [Stra8] and lower level of meiotic entry markers [SCP1, SCP3, and REC8] in juvenile than newborn mouse ovaries

Colony forming unit endothelial cells do not exhibit telomerase alternative splicing variants and activity

Endothelial progenitor colony forming unit-endothelial cells [CFU-EC] were first believed to be the progenitors of endothelial cells, named endothelial progenitor cells. Further studies revealed that they are monocytes regulating vasculogenesis. The main hindrance of these cells for therapeutic purposes is their low frequency and limited replicative potentials. This study was undertaken to determine telomerase activity and alternative splicing variants in CFU-EC as a potential cause of limited replicative capacity in these cells. CFU-EC were isolated from peripheral blood using a standard cell culture assay. Colonies were detached mechanically and alternative splicing variant mRNA were evaluated using real-time PCR. Telomerase enzyme activity was assessed using telomerase repeat amplification protocol. The same procedures were done on the cancer cell line Calu6 as the positive control. The cultured peripheral blood mononuclear cells formed colonies with spindle-shaped monocytic cells sprouted from the clusters. These morphological characteristics fulfill the definition of CFU-EC. Telomere length amplification protocol assay revealed no telomerase activity and real-time PCR showed no expression of telomerase enzyme mRNA in CFU-EC. Both parameters were significantly higher in the cancer cell line Calu6 taken as the positive control. The absence of telomerase activity in the CFU-EC is a result of pre-transcriptional regulation of gene expression rather than other mechanisms for controlling telomerase activity such as post-transcriptional modifications. This finding can explain the limited proliferative activity of CFU-EC cells. We propose that absence of telomerase activity in CFU-EC can be attributable to their more mature monocytic nature that needs further investigations

leishmania infantum fml-elisa for the detection of symptomatic and asymptomatic canine visceral leishmaniasis in an endemic area of Iran

Visceral leishmaniasis [VL] is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis [CVL]. In this survey a Fucose-Mannose Ligand [FML] ELISA, using native L. infantum antigen, was developed and its validity for detection of infected dogs in comparison with direct agglutination test [DAT] and PCR was evaluated. Blood samples of sixty ownership dogs [/= 1/320] in DAT while seven of the 60 [11.66%] samples were positive by FML-ELISA. Nine out of 60 [15%] buffy coat samples showed a band about 680 bp indicative of L. infantum in PCR. Three out of 60 dogs had Kala-azar symptoms and were positive by PCR and FML-ELISA, while two of these three dogs had antibody titers >/= 1/320 in their serum samples. The sensitivity and specificity of FML-ELISA for the detection of CVL in both symptomatic and asymptomatic dogs were found to be 77.8% and 100%, respectively. Considering the acceptable sensitivity and high specificity of FML-ELISA, use of this serological method can be recommended for epidemiological surveys of CVL