Chromosomal studies in infertile men

Prometaphase and metaphase chromosome analyses performed on 70 consecutive men with primary infertility (for a period of at least 2 years) revealed 8 (11.42%) men with some kind of chromosomal abnormality. The highest frequency of abnormal karyotypes (10%) was found among patients with azoospermia and the most frequent anomaly was 47, XXY chromosomal constitution, found in 6 (8.57%) patients. All the chromosomal aberrations found in this study, was sex chromosomal type and we did not find any autosomal aberration. All patients with numerical chromosomal anomalies had azoospermia. The incidence of structural aberration in our study was 1.42%. 15 patients had different chromosomal variants (21.38%). We suggest that men with azoospermia should be considered for cytogenetic investigation and we report that "variants of the Y chromosome" have no influence on the sperm count (Million/ml) and fertility of men.     

NO-donating aspirin inhibits intestinal carcinogenesis in Min (APC(Min/+)) mice

The chemopreventive effect of nitric oxide-releasing aspirin (NO-ASA) against gastrointestinal tumorigenesis was evaluated in Min (APC(Min/+)) mice. NO-ASA consists of a traditional ASA that bears covalently attached to it an NO-releasing moiety. Four groups (N=10) of six-week-old female C57BL/6J APC(Min/+) and the corresponding C57BL/6J(+/+) wild type mice were treated either with vehicle or NO-ASA 100 mg/kg/day intrarectally for 21 days. There were no signs of overt toxicity including gastrointestinal toxicity from NO-ASA. Vehicle treated Min mice had 24.7 +/- 3.8 tumors (mean +/- SEM) and NO-ASA treated Min mice had 10.1 +/- 1.4 tumors (59% reduction; P<0.001). Wild type mice showed no tumors. NO-ASA did not affect cell proliferation in small intestinal mucosa, determined by immunohistochemical staining for PCNA. Our findings establish the strong inhibitory effect of NO-ASA in intestinal carcinogenesis in the Min mouse and suggest that this agent merits further evaluation as a chemopreventive agent against colon cancer.     

Expression of cathepsin B and aryl hydrocarbon hydroxylase activities,and of apolipoprotein B in human hepatoma cells maintained long-term in a serum-free medium

Summary We have established the human hepatoma cell line, HepG2, in a defined, serum-free medium. These cells were maintained and studied over a 100-generation period (i.e. 10 serial transfers). Cells maintained in serum-free medium exhibited growth parameters (i.e. saturation density, efficiency of plating, and population doubling time) similar to those obtained with HepG2 cells maintained in serum-supplemented medium. Serum-free cells were also similar to their serum-supplemented counterparts with respect to the expression of cathepsin B activity and the induction of aryl hydrocarbon hydroxylase by 2,3,7,8-tetra-chlorodibenzo-p-dioxin. Significantly, HepG2 cells maintained in serum-free conditions also retained the ability to synthesize and secrete proteins, including the liver plasma protein, apo-lipoprotein B. These results indicate that the serum-free medium used in this study supports the long-term growth and maintenance of human hepatoma, HepG2, cells in culture. Inasmuch as these cells retain phenotypes, including differentiated markers previously reported for their serum-supplemented counterparts, they may provide a more reliable, standardized culture system to study the expression, secretion, and regulation of proteins during biological and pathologic processes.     

Synthesis, characterization and equilibrium study of the dinuclear adducts formation between nickel(II) Salen-type complexes with diorganotin(IV) dichlorides in chloroform

Adduct formation between R₂SnCl₂ (R = methyl and n-butyl) as acceptors, and nickel(II) complexes of tetradentate Schiff base ligands ([NiL]) where L = [3-methoxysalen, N,N′-bis(3-methoxysalicylidene)ethylenediamine], [4-methoxysalen, N,N′-bis(4-methoxysalicylidene)ethylenediamine], [5-methoxysalen, N,N′-bis(5-methoxysalicylidene)ethylenediamine], [salen, N,N′-bis(salicylaldehydo)ethylenediamine], [5-chlorosalen, N,N′-bis(5-chlorosalicylidene)ethylenediamine] and [5-bromosalen, N,N′-bis(5-bromosalicylidene)ethylenediamine] as donors have been investigated in chloroform as a solvent by means of UV-Vis spectrophotometeric analysis. Adducts have been characterized by ¹H NMR, IR and electronic spectroscopy. The formation constants and the thermodynamic free energies were measured using UV-Vis spectrophotometry titration for 1:1 adduct formation at various temperatures (T = 278-308 K). The trend of the adduct formation of the nickel Schiff base complexes with a given tin acceptor decreases as follow:

Ni(3-MeOSalen)>Ni(5-MeOSalen)>Ni(4-MeOSalen)     

Efficacy of Fe3O4/Starch Nanoparticles on Sporosarcina pasteurii Performance in MICP Process

The microbial induced calcite precipitation (MICP) has been explored using well-known urease producer bacterium Sporosarcina pasteurii for many applications including soil stabilization. Urease enzyme hydrolyzes urea and in the presence of calcium chloride causes calcium carbonate precipitation between sand particles increasing sand stiffness and strength. In this study, the liquefied soil samples from Anzali coast were positioned inside injection columns by standard positioning technique. The columns were treated by injecting S. pasteurii suspension and cementation solution (CaCl₂ and urea). The effect of different conditions consisting of number of injections, injection intervals, flow rate, and ratio of injection solution on unconfined compression strength (USC) of sands formed inside the columns were evaluated. The results indicated that soil strength was increased when ratio of reactant solutions and injection time were elevated. Moreover, the maximum Ca-precipitation in MICP reaction in liquid medium was obtained while Fe₃O₄/starch concentration and time of addition of nanoparticle to culture medium were 10.8?mg/L and 1.4?h, respectively. The USC results showed that the columns injected by bacterial suspension treated by Fe₃O₄/starch under optimized conditions improved the soil strength up to 1200?kPa in comparison to the control column as 220?kPa.     

Translational control of apolipoprotein B mRNA via insulin and the protein kinase C signaling cascades: evidence for modulation of RNA-protein interactions at the 5'UTR

The link between hepatic insulin signaling and apolipoprotein B (apoB) production has important implications in understanding the etiology of metabolic dyslipidemia commonly observed in insulin-resistant states. Recent studies have revealed important translational mechanisms of apoB mRNA involving the 5' untranslated region (5'UTR) and insulin-mediated translational suppression via an insulin-sensitive RNA binding protein. Here, we have investigated the role of the protein kinase C (PKCs) signaling cascade in the regulation of apoB mRNA translation, using a series of chimeric apoB UTR-luciferase constructs, in vitro translation of UTR-luciferase cRNAs, and metabolic labeling of intact HepG2 cells. The PKC activator, phorbol 12-myristate 13-acetate (PMA), increased luciferase expression of constructs containing the apoB 5' UTR whereas treatment with Bis-I, a general PKC inhibitor or Go6976, a more specific PKC alpha/beta inhibitor, decreased expression, under both basal and insulin-treated conditions. These effects were confirmed to be translational in nature based on in vitro translation studies of T7 apoB UTR-luciferase constructs transcribed and translated in vitro in the presence of HepG2 cytosol treated with insulin or signaling modulators. Mobility shift experiments using cytosol treated with either PKC inhibitor (Bis-I) or activator (PMA) showed parallel changes between translation of apoB 5'UTR-luciferase constructs and the binding of a protein(s) complex migrating around 110 kDa to the apoB 5' UTR. ApoB mRNA levels were unaltered under these conditions based on real-time PCR analysis. Bis-I and Go6976 were both able to significantly decrease newly synthesized apoB100 protein in the presence or absence of insulin. Overall, the data suggests that PKC activation may induce increased mRNA translation and synthesis of apoB100 protein through a mechanism involving the interaction of trans-acting factors with the apoB 5'UTR. We postulate potential links between PKC activation as seen in insulin-resistant/diabetic states, enhanced translation of apoB mRNA, and hepatic VLDL-apoB overproduction.     

Metastatic ganglioneuroblastoma in head and neck diagnosed by fine needle aspiration: a case report

BACKGROUND: To describe the cytologic findings of a case of ganglioneuroblastoma metastatic to the jaw and neck. CASE: A 15-year-old boy with a known case of ganglioneuroblastoma of the kidney for the previous 10 years manifested by right mandibular and neck masses on 2 occasions 1 year apart was diagnosed with metastatic ganglioneuroblastoma by fine needle aspiration (FNA). FNA showed neurofibrillary material, small malignant cells, Homer-Wright rosettes. mononucleated, binucleated and multinucleated ganglion cells and Reed Sternberg-like ganglion cells. Metastatic ganglioneuroblastoma was diagnosed on both occasions, and the patient received appropriate treatment, with resolution of the lesions. CONCLUSION: This case illustrates the FNA findings of metastatic ganglioneuroblastoma in the head and neck region.     

“Conservation cloning” of vulnerable Esfahan mouflon (Ovis orientalis isphahanica): in vitro and in vivo studies

Among the wide range of bio-conservational strategies envisaged, recent accomplishments in the field of interspecies somatic cell nuclear transfer (iSCNT) hold considerable promise due to its unique potential to decelerate or prevent rapid loss of animal genetic resources, and even to revive extinct species. Accordingly, this study was carried out to investigate if in vitro matured and enucleated oocytes of domestic sheep could be used for interspecies conservation cloning of Esfahan mouflon (Ovis orientalis isphahanica), a vulnerable species classified by the International Union for Conservation of Nature. Cryo-banked fibroblasts of a mouflon (derived from a genome resource bank) and a domestic sheep (prepared during a recent study) were cultured in vitro and used for karyotyping. Prior to SCNT, fibroblast donor cells were serum starved for 5 days. Using the zona-free SCNT technique, in vitro matured and enucleated domestic sheep oocytes were reconstituted with nuclei donor cells of mouflon and domestic sheep, and their competencies for in vitro development to the blastocyst stage were compared. The cloned mouflon blastocysts were then surgically transferred into the uterus of the synchronized domestic sheep. Karyotype analysis confirmed that fibroblasts of the Esfahan mouflon had the correct number of diploid chromosomes (2n = 54). Evaluation of 907 activated reconstructs [Esfahan mouflon (n = 667), domestic sheep (n = 240)] revealed no significant difference in the term of blastocyst development (7.6 ± 0.5% vs. 9.3 ± 0.5%, respectively). After the transfer of 12 cloned Esfahan mouflon blastocysts to five domestic sheep recipients, two (40.0%) pregnancies were established in which both (100%) were sustained until caesarean section (days 147 and 150 of pregnancy, respectively) and culminated in the live births of cloned Esfahan mouflon lambs. However, the newborns did not survive and died soon after birth. Karyotype and genetic analyses confirmed that both clones had correct diploid chromosome number (2n = 54), and were genetically identical to each other in addition to their original cell donor. This study highlighted the importance of “conservation cloning” using closely related abundant alternate species.