KALLIKREINS AND KININS IN INFLAMMATORY-LIKE EVENTS IN THE REPRODUCTIVE TRACT

The normal reproductive events of proliferation of the endometrial lining of the uterus during the menstrual cycle and ovulation have been likened to inflammatory-like events. The kallikrein-kinin system is involved in inflammatory processes in many tissues. In this review, we identify which components of the kallikrein-kinin system — the enzyme, tissue kallikrein; the substrate, low molecular weight kininogen and the effector receptor for the generated bradykinin peptide, the B2 receptor — have been identified in the uterus and ovary and their known involvement in the function of these organs. All three components have been localized to the glandular epithelial cells of the human endometrium. Tissue kallikrein gene expression is elevated midcycle when estrogens levels are also rising. This is also a time of extensive endometrial proliferation and tissue remodelling in preparation for embryo implantation, an event which is likened to other inflammatory processes. Similarly, tissue kallikrein gene expression was elevated following the estrogen surge at proestrous in the rat uterus, suggesting tissue kallikrein gene expression may be regulated by estrogens. Tissue kallikrein enzyme activity and gene expression has been demonstrated in the rat ovary and shown to be variously altered at the time of ovulation. Bradykinin has also been implicated in the expulsion of the ovum at the time of ovulation. These findings show that various components of the kallikrein-kinin system are present in the uterus and ovary. Further studies are required to more fully delineate their role in reproductive function.     

Herpes simplex virus type‐2 in Egyptian patients with bladder cancer or cystitis

Badawi H, Ahmed H, Aboul Fadl L, Helmi A, Fam N, Diab M, Ismail A, Badawi A, Saber M. Herpes simplex virus type‐2 in Egyptian patients with bladder cancer or cystitis. APMIS 2010; 118: 37–44. The present study was designed to investigate the prevalence of herpes simplex virus type‐2 (HSV‐2) in Egyptian patients with bladder cancer or cystitis and to evaluate the performance of different diagnostic HSV‐2 assays. The study included 50 patients: 27 with bladder cancer (group I), 23 with cystitis (group II) and 20 subjects as controls (group III). HSV‐2 DNA was detected using polymerase chain reaction (PCR) on bladder tissue and buffy coat cells (BCC). Electron microscopic studies (EMS) on BCC and ELISAs for IgM, IgG and specific glycoprotein G‐2 (gG‐2) IgG were performed. HSV‐2 DNA was detected by PCR on bladder tissue biopsies in 29.6% and 21.7% of group I and II respectively and it was also detected by PCR on BCC in 22.2% and 21.7% of group I and II respectively. EMS revealed HSV like particles in 16.6% of cases. IgG, specific gG‐2 IgG and IgM were detected in 30%, 16% and 6% of cases respectively. The different assays were evaluated in relation to PCR on bladder tissue biopsies. The gG‐2‐based ELISA and EMS on BCC were found to be highly specific (97.3% and 100% respectively), with similar low sensitivity of ≈54%. PCR on BCC was the most sensitive assay. The association of HSV‐2 with bladder cancer is suggested especially in schistosomal patients.     

HLA ANTIGENS AND THYROID AUTOANTIBODIES IN PATIENTS WITH GRAVES' DISEASE AND THEIR FIRST DEGREE RELATIVES

Patients with Graves' disease (n= 105) had an increased frequency of HLA-B8 (40%) and a reduced frequency of HLA-B12 (24·8%) when compared with random controls (n=117; 24·8% and 40·2% respectively). Comparison of patients with their first degree relatives (n= 118) shows the frequency deviations in these antigens to be characteristic of the families from which patients with Graves' disease are drawn, rather than of the disease itself. The haplotypes, identified in eighty-six patients and 113 relatives, indicate that the excess of HLA-B8 in patients and their relatives is primarily due to the haplotype 1–8. The relative risk for an HLA-B8 individual of developing Graves' disease is 2·02, whilst the relative risk for an individual of haplotype 1–8 is 4·23. No significant associations were found between the incidence of any HLA antigen or combination thereof and the presence or absence of thyroglobulin and thyroid microsomal antibodies, or antibodies which interact with the TSH receptor.     

Suppression of Schistosoma bovis egg production in cattle by vaccination with either glutathione S-transferase or keyhole limpet haemocyanin

Two of the antigens which have shown vaccine potential in animal experiments against Schistosoma mansoni are glutathione-S-transferase (GST) and GP38, protective epitopes of which are shared with keyhole limpet haemocyanin (KLH). We therefore tested S. bovis GST and KLH for vaccine efficacy against S. bovis in the natural Zebu cattle host. In a preliminary experiment three vaccinations with a total of 1.39 mg of native GSTs of S. bovis induced specific antibody at the time of challenge as detected by Western blotting and ELISA and mean faecal egg counts between weeks 6-10 post-challenge were reduced by 56.4 to 82.5% compared to non-vaccinated controls. Mean adult worm recoveries and tissue egg densities in large intestine and liver samples were also reduced in the vaccinated group, but these differences were not statistically significant. In a subsequent experiment one group of calves was vaccinated with a similar schedule to that used above; a second group of calves was given only two injections of GST (total 0.48 mg protein); a third group of calves was vaccinated twice with a total of 2.0 mg KLH in PBS. All three vaccination schedules induced specific antibody. Both GST vaccination schedules induced significant reductions in faecal egg counts compared to non-vaccinated controls and in this experiment tissue egg densities were also significantly reduced. A striking finding, however, was that adult worm counts were not reduced by vaccination. An essentially similar outcome resulted from KLH vaccination, since there were significant reductions in faecal and tissue egg counts in the absence of a reduction in adult worm numbers. This type of immunity mimics that induced by natural or experimental infections in the calf and clearly has implications for vaccine design.     

Benzo(a)pyrene metabolism, DNA-binding and UV-induced repair of DNA damage in cultured skin fibroblasts from a patient with unilateral multiple basal cell carcinoma

The metabolism of benzo(a)pyrene (BP), a ubiquitous environmental carcinogen, and its subsequent binding to DNA, and the repair of UV-induced DNA damage were studied in fibroblasts cultured from the skin of a 61-year-old male who had multiple BCC (greater than 100) on his left upper trunk. Biopsies were obtained and fibroblasts cultured from clinically normal tumour-free skin adjacent to tumour-bearing sites (TBS) and from visibly uninvolved normal skin (UNS) at distant sites. The cultured cells were incubated with [3H]-BP for 24 h and BP metabolism was assessed by HPLC and the formation of BP-diols, quinones and phenols verified. Total BP metabolism was 45% lower in TBS fibroblasts than in UNS fibroblasts. The formation of BP-7,8-diol, the precursor of the carcinogenic end product of BP metabolism, was 53% lower in TBS cells than in UNS cells. Pretreatment of UNS cells with benz(a)anthracene (BA) (x 10(-4) M) did not significantly affect BP metabolite formation whereas BA-treatment of TBS cells resulted in 55% and 76% increases in total BP metabolism and BP-7,8-diol formation, respectively. Treatment of TBS cells with BA also caused a substantial increase (95%) in BP-DNA adduct formation. Whereas DNA-binding in UNS cells was unaffected by this treatment. In response to irradiation with 2J/m2 UVC, total DNA repair was similar in both cell types; on alkaline elution it appeared that the TBS cells were more efficient in repairing UV-induced DNA strand breaks. These results suggest that BP metabolism and repair of DNA are altered in TBS cells and that patients with this type of metabolic profile may be at higher risk of the development of cutaneous neoplasms. It is also possible that fibroblasts from tumour bearing skin undergo some as yet unexplained alteration in carcinogen metabolism as a consequence of the induction of neoplasia.     

Lectin binding characteristics of mouse placental cells

The lectin binding characteristics of mouse placental cells were examined. Wax embedded tissue sections of placentae from d 14 pregnant mice were stained with 26 lectins, with a wide range of sugar specificities. Cell cultures prepared from d 14 mouse placentae and cultured for 24 h were stained with 7 of the lectins to determine if they could be used as markers for the different trophoblast cells in culture. In tissue sections all placental cell populations bound lectin but no lectin bound specifically to any single trophoblast population. All the lectins which bound to layer 1 cytotrophoblast lining the maternal blood spaces of the labyrinthine placenta also bound to the fetal endothelium of the labyrinthine placenta. Binding of lectin appeared strongest on the adluminal membrane of these cell populations suggesting a role for the carbohydrate moieties in nutrient transfer. Few lectins bound to junctional zone trophoblast. Overall, the binding of lectin to cultured cells did not correlate exactly with lectin binding to the cell populations in tissue sections. The value of lectins as markers for placental cells in culture was therefore found to be limited. Our findings indicate that carbohydrate expression by at least some placental cells may vary in culture from that expressed by the cells in vivo with obvious concerns for the validity of functional in vitro studies.