A conundrum in molecular toxicology: molecular and biological changes during neoplastic transformation of human cells

The process of multistage carcinogenesis lends itself to the concept that the effects of carcinogens are mediated through dose-related, multi-hit, linear changes. Multiple in vitro model systems have been developed that are designed to examine the cellular changes associated with the progression of cells through the different stages in the process; however, these systems may have inherent limitations due to the cell lines used for these studies, the manner of assessing the effects of the carcinogens, and the subsequent growth and differentiation of the exposed cells. Each of these variables results in increasing levels of uncertainty relative to the correlation of the events with the actual process of human tumor development. Therefore, the prediction of the ultimate effect of any carcinogen is difficult. Moreover, relationships between individual biological endpoints resulting from carcinogen treatment appear at best to be approximations. The presence of an activated carcinogen inside the cell can give rise to multiple outcomes, only some of which may be critical events. For example, site-specific modification of the 12th and 13th codons of H-ras is different than that in the adjacent 14th and 15th codons. It is interesting to speculate what effect these differences might have on a biological outcome, e.g., transformation to anchorage-independent growth. The use of different model systems to examine the effects of activated carcinogens also creates additional problems. Comparisons of in vitro transformed cells with similar cells isolated from human tumors indicate that the culture environment appears to influence the expression of a particular phenotype, in that human tumor cells in culture express many of the same parameters as those found in cells transformed with carcinogens in vitro. If the process of transformation is linear, then less aggressive phenotypes should progress to a more aggressive transformed stage. However, in carcinogen-transformed human cells, the populations exhibit phenotypic diversity in that many of the transformed cells differentiate and fail to continue to divide in culture. Historically, we have assumed only a limited role for epigenetic modulation of molecular changes that occur during progression; however, our data suggest quite strongly that nonmalignant tumor populations can be converted to a more malignant phenotype without additional mutations taking place and, conversely, malignant populations can be downregulated to a nontumorigenic phenotype. Tumor cell plasticity is not only a fundamental characteristic of diverse types of human tumors, but also appears as an integral characteristic of carcinogen-transformed cells in vitro.     

A study of female urinary incontinence in general practice. Demography, medical history, and clinical findings

The aim of this study was to provide valid data on the demography, medical history and clinical findings among adult women presenting with urinary incontinence to general practitioners. In a rural community in Norway, all women > 20 years who consulted their general practitioner for urinary incontinence during a 3 year period were included in a prospective study. A thorough medical history and both a general and focused clinical examination were undertaken. Gynecological examination, stress provocation test, and 48 h frequency/volume chart and pad weighing test were also performed. 105 women were included (4.4% of women > 20 years in the total population). Mean age was 57 years, 64% were postmenopausal. A lot of comorbidity was reported. Duration of incontinence was > 5 years in 49%. By a severity index, 64% were classified as severe, 28% as moderate and 8% as having slight incontinence. 59% were using protective pads or garments. Mean leakage per 24 h was 31 g. 38% had significant genital prolapse. Contractility of the pelvic floor muscles was weak in 28%. Diagnostic classification revealed 50% stress incontinence, 10% urge and 40% mixed incontinence. 42% of the patients were a great deal or much bothered by their incontinence. Patients with stress incontinence were less bothered than others. Women presenting with urinary incontinence at a primary care level are prevalent, and often have significant incontinence. It is a challenge for the general practitioners to investigate and treat these patients optimally.     

Plasma LH progesterone concentrations in the turkey hen during the ovulatory cycle

Progesterone and luteinizing hormone (LH) were measuured in the plasma of Large White turkey hens at frequent intervals during the ovulatory cycle and during the periods when ovulations did not occur. The hormones were quantitated at the beginning and the end of the reproductive season to follow the change in patterns and concentrations of these hormones. Radioimmunoassay procedures were used to assay both hormones. Both progesterone and LH reached a peak amount at about 8 to 2 hours before ovulation. At no time did the progesterone peak precede that of LH. On the other hand, the peak of progesterone was observed to last somewhat longer than the LH peak. Progesterone and LH concentrations demonstrated no peak during the non-ovulatory periods. The patterns of both hormones at the end of the reproductive season was about the same as at the beginning of the season, but with lower concentrations.     

Conservation of the centromere/kinetochore protein ZW10

Mutations in the essential Drosophila melanogaster gene zw10 disrupt chromosome segregation, producing chromosomes that lag at the metaphase plate during anaphase of mitosis and both meiotic divisions. Recent evidence suggests that the product of this gene, DmZW10, acts at the kinetochore as part of a tension-sensing checkpoint at anaphase onset. DmZW10 displays an intriguing cell cycle-dependent intracellular distribution, apparently moving from the centromere/kinetochore at prometaphase to kinetochore microtubules at metaphase, and back to the centromere/kinetochore at anaphase (Williams, B.C., M. Gatti, and M.L. Goldberg. 1996. J. Cell Biol. 134:1127-1140). We have identified ZW10-related proteins from widely diverse species with divergent centromere structures, including several Drosophilids, Caenorhabditis elegans, Arabidopsis thaliana, Mus musculus, and humans. Antibodies against the human ZW10 protein display a cell cycle-dependent staining pattern in HeLa cells strikingly similar to that previously observed for DmZW10 in dividing Drosophila cells. Injections of C. elegans ZW10 antisense RNA phenocopies important aspects of the mutant phenotype in Drosophila: these include a strong decrease in brood size, suggesting defects in meiosis or germline mitosis, a high percentage of lethality among the embryos that are produced, and the appearance of chromatin bridges at anaphase. These results indicate that at least some aspects of the functional role of the ZW10 protein in ensuring proper chromosome segregation are conserved across large evolutionary distances.     

Nitric oxide in renal health and disease

Nitric oxide (NO) is a labile radical gas that is widely acclaimed as one of the most important molecules in biology. Through covalent modifications of target proteins and redox reactions with oxygen and superoxide radical and transition metal prosthetic groups, NO plays a critical role in many vital biological processes, including the control of vascular tone, neurotransmission, ventilation, hormone secretion, inflammation, and immunity. Moreover, NO has been shown to influence a host of fundamental cellular functions, such as RNA synthesis, mitochondrial respiration, glycolysis, and iron metabolism. NO is formed from L-arginine by NO synthases (NOSs), a family of related enzymes encoded by separate unlinked genes. The different NOS isozymes exhibit disparate tissue and intrarenal distributions and are governed by unique regulatory mechanisms. In the kidney, NO participates in several vital processes, including the regulation of glomerular and medullary hemodynamics, the tubuloglomerular feedback response, renin release, and the extracellular fluid volume. While NO serves beneficial roles as a messenger and host defense molecule, excessive NO production can be cytotoxic, the result of NO's reaction with reactive oxygen and nitrogen species, leading to peroxynitrite anion formation, protein tyrosine nitration, and hydroxyl radical production. Indeed, NO may contribute to the evolution of several commonly encountered renal diseases, including immune-mediated glomerulonephritis, postischemic renal failure, radiocontrast nephropathy, obstructive nephropathy, and acute and chronic renal allograft rejection. Moreover, impaired NO production has been implicated in the pathogenesis of volume-dependent hypertension. This duality of NO's beneficial and detrimental effects has created extraordinary interest in this molecule and the need for a detailed understanding of NO biosynthesis.