Iranian Biomedical Journal
Volume:2 Issue: 1, Jan 1998

Three tuberculous, twenty-one non-tuberculous mycobacterial (NTM) reference strains and seventy two isolates classified by biochemical tests were shown to produce specific sets of DNA fragments in a polymerase chain reaction with single universal primer (UP-PCR). A rather wide limit of tolerance for variations in procedure of PCR mixture preparation and thermocycling parameters was found. There was good correlation between UP-PCR pattern obtained in an agarose gel for each strain and the absolute majority of isolates tested. The PCR products of M. tuberculosis and M. bovis were shown to be very similar and well distinctive from that of all NTM strains tested. When image analy‌sis system was applied to profile the UP-PCR patterns, all M. tuberculosis complex isolates were well differentiated from NTM species according to their specific profiles. Thus, UP-PCR profiling could give an efficient means for discriminating mycobacteria that utilizes the least time and applicable to controversial cases of suspected tuberculosis.

Antiproliferative protein (APP) isolated from conditioned media of two androgen-independent prostatic carcinoma cell lines, PC3 and Du-145 was shown to inhibit selectively cell proliferation of androgen-dependent prostate cancer cell line LNCaP in a dose dependent manner. This protein was further purified with HPLC using hydrophobic interaction column (phenyl 5PW) and was used to study the modulation of protein phosphorylation of LNCaP cells. The data indicated that antiprolif-erative protein could partially change the cytosolic protein phosphorylation. When compared with control LNCaP cells, in APP-treated cells 4 new proteins (88, 46, 30 and 18 kDa) were phosphory-lated, while a 72 kDa phosphoprotein was de-phosphorylated. The same results were obtained when two types of protein kinase activators were used. Protein kinase activators showed that the above changes of protein phosphorylation are not due to the protein kinase C or DNA protein kinase activ‌ity. These data may indicate the inhibition of LNCaP cell's proliferation by APP may be is due to the modulation of protein kinases and as a result due to interference on second messenger pathway.

In vitro lymphocyte responses of high responder (HR) and low responder (LR) guinea pigs from pe-ripheral blood lymphocytes (PBL) to parasite antigens soluble protein third stage larvae (SPL3) and soluble protein adult stage (SPA), non-parasite antigen ovalbumin (OVA) were examined. There was substantial differences between HR and LR guinea pigs in the rate of acquisition of responsiveness to these Ags as well as differences in responsiveness to Ags derived from third stage larvae (SPL3) and adult worms (SPA). Overall, the results suggest both stronger and more rapid acquisition of respon‌siveness by HR animals and raise the possibility of the animals being able to preferentially respond to larval immunogen and thus acquire protective immunity more rapidly than LR.

Four exons of the human genomic GM-CSF gene were assembled together using gene splicing by overlap extension (SOE) method. The resulting nucleotide sequence was cloned in the pET23a(+) ex‌pression vector under the control of strong bacteriophage T7 transcription and translation signals. The construct obtained was Transferred into the E. coli strain, BL21(DE3) pLysS and IPTG was used for induction of GM-CSF gene and production of the target protein. one mg of protein per liter of cell culture, was obtained as revealed by ELISA.

Partially purified antigenic fractions of Leishmania major promastigotes were obtained by sodium dodecyl sulfate (SDS-PAGE) and electro-elution. Three isolated protein fractions designated as frac tions (Fr.) 1, 2, and 3 correspond to 40-60, 60-80, 80-100 kilodalton (kDa) respectively. They were used for immunization of BALB/c mice against Leishmaniasis. The effects of these fractions on immune response of BALB/c mice against leishmanial infections was investigated by studying the infection course in infected mice, delayed type hypersensitivity (DTH) skin test, lymphocyte proliferation assay (LTT) in them. Subcutaneous immunization of mice with fraction 2 in conjugation with Complete Freund's Adjuvant (CFA) developed partial immunity against Leishmania major infection, and in duced specific DTH response. Meanwhile this fraction exhibited no exacerbating effect on leishmanial infection course. Subcutaneous immunization with fraction 1 also induced partial protection in lesser extent than fraction 2 against leishmanial infection.

It is now well established that the type la of serine/threonine protein phosphatases (PP1α) may be implicated in malignant phenotype. Although the PP1α mRNA level increased in livers at preneo-plastic stages of hepatocarinogenesis and all of examined rat ascites hepatomas, unexpectedly, dra-matically decreased in some of primary hepatomas. In order to elucidate the low level of PP1α mRNA in some of primary hepatomas and investigate any correlation between cell growth rates and change(s) of both PP1α mRNA and PP1 activity, the present experiments were done. The most im‌portant aspects of the present study are:'' 1) The PP1α mRNA level may decreased in ascites hepa‌toma AH-109A cells harvested from a highly bloody ascites compared to ones harvested from a milky ascites. 2) The spontaneous PP1 activity in particulate fraction of AH-7974F showed a positive corre‌lation with cell growth rate. 3) There was no correlation between proliferation rate of AH-7974F and the amount of PP1α catalytic subunit as well as potential PP1 activity in particulate fraction. There‌fore, it is suggested that the PP1α mRNA level may decreased in the primary hepatomas with low proliferation rate which were under very bad nutritional conditions.

The effect of anti-follicular stimulating hormone (FSH) on testicular lipid and the specific activity of testicular enzymes of the isocitrate dehydrogenase (ICDH), pyruvate/malate dehydrogenase (MDH) and malic enzyme involved in lipogenesis were studied in mature bonnet monkeys, Macaca radiata. Immunization of monkey with anti-FSH for 24 days did not produce any significant changes in the body weight, organ weight and pituitary weight. Testicular isocitrate dehydrogenase (ICDH) and malic enzyme activities were decreased significantly but MDH activity was stimulated by anti-FSH treatment. Testicular total lipid, phospholipid and cholesterol, were not altered significantly by the Anti-FSH treatment. Increased level of free cholesterol was also observed after FSH treatment. Among glyceride glycerol sub classes, triacyl glycerol showed a significant increase.Among testicular phospholipid classes, phosphatidyl inositol was markedly decreased by anti-FSH immunization. Data on serum hormonal profile, shows that there were no alteration in serum testosterone, prolactin (PRL) and luteinizing hormones (LH) but FSH was significantly decreased. The present study reveals that immunization with anti-FSH has significant effect on different class Of testicular lipids and pyruvate malate enzymes cycle.

Chlamydia trachomatis is an obligate intracellular bacterium which causes a wide variety of human infections such as ocular, urogenital and respiratory infections. Genital infections of women, espe-cially when repeated, give rise to many complications such as ectopic pregnancy, miscarriage and in‌fertility. Since chlamydial infections are usually asymptomatic, they progress unnoticed and produce the sequela. The method of choice for the isolation of C. trachomatis is cell culture but other tech‌niques like DIF and ELISA are more feasible, faster, less expensive and adequately specific and sensi‌tive. In this study, prevalence of C. trachomatis antigen and antibody was assessed in a high risk group of infertile women by DIF and ELISA methods in Ahwaz, Southwest of Iran. The rates obtained were much less than those of many other populations, e. g. USA, Sweden, Australia, South Africa and Tur‌key. It was also concluded that presence of C. trachomatis antigen and antibody was associated with infertility in these women when compared with healthy pregnant control women.