Cytokeratin 14 expression in rat liver cells in culture and localization in vivo

Rat liver epithelial cells (LECs) are non-parenchymal proliferating cells that readily emerge in primary culture and can be established as cell lines, but their in vivo cell(s) of origin is unclear. We reported recently some evidence indicating that the LEC line, T51B, contains two cytokeratins (CKs) equivalent to human CK8 and CK14 respectively. T51B cells also contain vimentin assembled as a network of intermediate filaments distinct from that of the CKs. In the present study, we examined the expression of CK14 gene in various LEC preparations and a Triton-resistant rat skin cytoskeletal fraction, and then assessed its usefulness as an LEC specific marker in the liver. Northern and Western blot analyses with cDNAs and antibodies for CK8, CK14, CK18 and vimentin confirmed that rat hepatocytes express CK8 and CK18 genes only, whereas T51B cells express CK8, CK14 and vimentin genes in the absence of CK18. CK14 was also present in LECs derived as primary from embryonic-day 12 rat liver and secondary cultures from 4-day-old rat liver. Primary cultures of oval cells isolated from 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) treated rat liver (an enriched source of biliary epithelial cells) contained CK14 mRNAs which were slightly shorter than those in LECs. The analyses of CK5 (the usual partner of CK14) gene expression using specific cDNA and antibody clearly demonstrated its absence in LECs. In situ double immunolocalization analyses by laser scanning confocal microscopy showed that CK14 was not present in hepatocytes (HES6+ cells) and was expressed in some biliary epithelial (BDS7+ cells). CK14-positive cells were also found in the Glisson's capsule. However, CK14-positive cells of the portal region were vimentin negative, whereas those of the Glisson's capsule were vimentin positive. Our results suggest that CK14 gene expression is part of the differentiation program of two types of LECs and that this differential CK14 gene expression can be used as a new means to type LECs in culture and in vivo.     

Contribution de la testostérone biodisponible aux explorations hormonales pour le diagnostic de l’andropause

The measurement of bioavailable testosterone (BT) is considered to be an essential analytical criterion for the diagnosis of male hypogonadism, but the reported normal values differ from one study to another and no consensus has been reached concerning the cut-off values for the diagnosis of androgen deficiency in aging males. Using the lower values measured in a group of clinically normogonadic men between the ages of 40 and 49 years, we have established our own cut-off values: 8 nmol/L for total testosterone and 3.5 nmol/L for BT. By applying these criteria to a group of 87 men with clinical symptoms of androgen deficiency, androgen deficiency was confirmed by laboratory assays in only 14% of these men based on total testosterone, but in 60% of men based on BT. The inverse correlation between BT and SHBG was confirmed regardless of the total testosterone level. SHBG assay is very useful for the diagnosis of androgen deficiency in a population of older males, but cannot replace direct measurement of BT for an accurate individual diagnosis.     

Long-term maintenance of hepatocyte functional activity in co-culture: requirements for sinusoidal endothelial cells and dexamethasone

Sinusoidal cells isolated from adult rat liver have been established in primary culture and in cell line. The presence of factor VIII R:Ag and peroxidatic/phagocytosis activities were the criteria used to distinguish in freshly isolated cells the endothelial cells from the Kupffer cells and suggested the endothelial origin of the cell line. Using a co-culture system, the effect of sinusoidal liver cells on hepatocyte functional activity was characterized. A plateau in which the state of differentiation was stabilized could be generated for co-cultured hepatocytes isolated from adult rat and a disappearance of the initial expression of alpha 1-fetoprotein (AFP) and the increase and/or maintenance of albumin secretion were measured with co-cultured hepatocytes isolated from suckling rat. The presence of dexamethasone was required for such beneficial effect. The hepatocyte-stabilizing activity was also produced by a pulmonary endothelial cell line.     

Contribution of hGH20K variant to blood hGH response in sauna and exercise

Exercise-induced increases in blood somatotropin (hGH) have always been considered in terms of quantity of the circulating molecules. Knowing that the hypophysis can release several GH species, we investigated the differential release in blood of total hGH (hGHT) and the main hGH variant (hGH20K) molecules in six trained male swimmers exposed to three different conditions known to favor GH release in blood: 45 min--70% maximum oxygen uptake (VO2max) bicycling and swimming, and 20 min of sauna bathing. Based on the binding specificity of hGH antibodies, hGH20K was isolated then assayed using the Nichols immunoradiometric assay system. All three experimental conditions produced significant (P less than 0.001) elevations in blood hGHT and hGH20K. In all three cases, mean blood hGH20K contribution to blood hGHT was relatively constant (11.9, SE 0.7%). Rises in rectal temperature were not statistically related to the changes in blood hGHT. This demonstration of a relatively constant elevation in hGH20K during bicycling, swimming, and sauna bathing can hardly explain the large differences in blood hGHT responses reported in literature under similar conditions.     

Family violence: guidelines for recognition and management

Chronic and intermittent abuse of one family member by another is common. Victims may be children who are sexually or physically abused, wives or live-in partners, or older relatives. Physicians are often the first points of contact for patients who have been abused, but the abuse is frequently concealed by the victims. Physicians should be alert to signs of battering such as bruises in various stages of healing, unusual behaviour in children and interpersonal difficulties in the family. There are a number of options in prevention and treatment, including referral to social service and legal authorities, calling on other resources in the family and helping the individual develop coping skills. This review also lists a large number of social agencies in Canada that are willing to help victims of abuse.     

Alkaline phosphatase activity at unusual sites of human foetal kidney

Summary Frozen sections of human foetal kidneys were treated by the lead citrate method in order to demonstrate alkaline phosphatase (ALP) activity. Unexpectedly, ALP activity was observed over the parietal layer of Bowman's capsule, in addition to usual localization in proximal tubules and blood vessels. The enzyme was confined to the membrane of microvilli belonging to tall columnar cells. Normally, in the mature nephron, these high cells are absent from the parietal layer that is instead entirely composed of squamous epithelium.ALP activity was also revealed at another unusual site in the kidney medulla. A collar of mesenchymal cells encapsulating groups of tubules and calyces gave an intense enzymatic reaction. The activity was present over the membrane of long cytoplasmic processes. The cytoplasm showed a well developed rough endoplasmic reticulum, an indication of a high rate of protein synthesis. The nearby presence of numerous collagenous fibrils is concordant with this assumption. Moreover, these ALP-positive cells might represent an intermediary stage through which cells have to pass before differentiating.     

Tn 5 to assess soil fate of genetically marked bacteria: screening for aminoglycoside-resistance advantage and labelling specificity

Abstract The reliability of Tn 5 as labelling tool was investigated in soil microcosm. The occurence of a selective in soil microcosm. The occurence of resistances encoded by Tn 5 nptII gene was assesed by kanamycin and neomycin amendment. The bioassay developed to monitor the persistence of the soil-added kanamycin did not detect the antibiotic activity in soil extract. A nptII -engineered Escherichia coli strain showed no enhanced survival in aminoglycoside amended soil. Tn 5-marker properties were investigated within indigenous bacteria to determine the specificity of labelling to follow the fate of recombinant DNA. Kanamycin and neomycin resistant population levels made Tn 5 aminoglycoside-resistance phenotype non-sensitive enough to select a soil dissemination of the labelled DNA. The unexpected occurrence of homologous sequences among soil organisms also prevented Tn 5 from being a specific DNA marker. By contrast, colony hybridization did not reveal homology to nptII suggesting its use as a reliable gene transfer indicator.