Mek2 is dispensable for mouse growth and development

MEK is a dual-specificity kinase that activates the extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinase upon agonist binding to receptors. The ERK/MAP kinase cascade is involved in cell fate determination in many organisms. In mammals, this pathway is proposed to regulate cell growth and differentiation. Genetic studies have shown that although a single Mek gene is present in Caenorhabditis elegans, Drosophila melanogaster, and Xenopus laevis, two Mek homologs, Mek1 and Mek2, are present in the mammalian cascade. The inactivation of the Mek1 gene leads to embryonic lethality and has revealed the unique role played by Mek1 during embryogenesis. To investigate the biological function of the second homolog, we have generated mice deficient in Mek2 function. Mek2 mutant mice are viable and fertile, and they do not present flagrant morphological alteration. Although several components of the ERK/MAP kinase cascade have been implicated in thymocyte development, no such involvement was observed for MEK2, which appears to be nonessential for thymocyte differentiation and T-cell-receptor-induced proliferation and apoptosis. Altogether, our findings demonstrate that MEK2 is not necessary for the normal development of the embryo and T-cell lineages, suggesting that the loss of MEK2 can be compensated for by MEK1.     

A study of the yeast cell wall composition and structure in response to growth conditions and mode of cultivation

AIM: The polysaccharide composition of the Saccharomyces cerevisiae cell wall was measured under various growth conditions and was compared with the cell wall structure. METHODS AND RESULTS: Chemical and enzymatic methods were used to determine levels of beta-1,3-glucan and 1,6-glucan, mannan and chitin of the yeast cell wall, whereas the structure/resistance of the wall was qualitatively assessed by the sensibility to the lytic action by zymolyase. It was found that the dry mass and polysaccharides content of the cell wall could vary by more than 50% with the nature of the carbon source, nitrogen limitation, pH, temperature and aeration, and with the mode of cell cultivation (shake flasks vs controlled fermentors). While no obvious correlation could be found between beta-glucan or mannan levels and the susceptibility of whole yeast cells to zymolyase, increase of beta-1,6-glucan levels, albeit modest with respect to the growth conditions investigated, and to a lesser extent that of chitin, was associated with decreased sensitivity of yeast cells to the lytic action by zymolyase. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results indicate that the cell wall structure is merely determined by cross-linking between cell wall polymers, pointed out the role of beta-1,6-glucan in this process. Hence, this study reinforces the idea that enzymes involved in these cross-linking reactions are potential targets for antifungal drugs.     

The eukaryotic nucleotide excision repair pathway

Nucleotide excision repair (NER) is the most versatile mechanism of DNA repair, recognizing and dealing with a variety of helix-distorting lesions, such as the UV-induced photoproducts cyclobutane pyrimidine dimers (CPDs) and pyrimidine 6-4 pyrimidone photoproducts (6-4 PPs). In this review, we describe the main protein players and the different sequential steps of the eukaryotic NER mechanism in human cells, from lesion recognition to damage removal and DNA synthesis. Studies on the dynamics of protein access to the damaged site, and the kinetics of lesion removal contribute to the knowledge of how the cells respond to genetic insult. DNA lesions as well as NER factors themselves are also implicated in changes in cell metabolism, influencing cell cycle progression or arrest, apoptosis and genetic instability. These changes are related to increased mutagenesis and carcinogenesis. Finally, the recent collection of genomic data allows one to recognize the high conservation and the evolution of eukaryotic NER. The distribution of NER orthologues in different organisms, from archaea to the metazoa, displays challenging observations. Some of NER proteins are widespread in nature, probably representing ancient DNA repair proteins, which are candidates to participate in a primitive NER mechanism.     

Lipid peroxidation and scavenging enzyme levels in the liver of streptozotocin-induced diabetic rats

In this study, alterations in the liver antioxidant enzymes status and lipid peroxidation in short-term (8-weeks) and long-term (24-weeks) diabetic rats were examined. Glutathione peroxidase (GSH-Px) activity and malondialdehyde (MDA) levels were significantly increased, but superoxide dismutase (SOD) activity was significantly reduced in 8-weeks diabetic rats, compared to control. Catalase (CAT) activity, however, was found unchanged. In 24-weeks diabetic rats, while GSH-Px activity was unchanged, but SOD and CAT activities and MDA levels were significantly increased, compared to control. These results suggest that diabetes-induced alterations in tissue antioxidant system may reflect a generalized increase in tissue oxidative stress. It can be concluded that lipid peroxidation and antioxidant enzyme levels are elevated in diabetic condition. Hence, diabetes mellitus, if left untreated, may increase degenerative processes due to accumulation of oxidative free radicals.     

Distribution of the intensity-characteristic parameters of cat rapidly adapting mechanoreceptive fibers

Modeling population responses of nerve fibers requires statistical characterization of fiber-response properties. The rate/intensity characteristics of cat rapidly adapting (RA) fibers were fitted by four-parameter, piece-wise linear functions using nonlinear regression (n = 14; R2 > 0.958). The parameters were tested against the null hypothesis that they are log normally distributed. The test fail to reject this hypothesis (Kolmogorov-Smirnov p>0.477). However, a significant statistical difference was found between the specific lognormal distributions obtained from monkey (Johnson, J Neurophysiol 37: 48-72, 1974) and cat for all four parameters (Kolmogorov-Smirnov, p<0.0075, p<0.05, p<0.0001, p<0.00007). Although the stimulus contactor size was not the same in monkey and cat studies, the differences between monkey and cat fibers are attributed to anatomical differences in the glabrous sin of both species. Modeling studies suggest that the absolute firing thresholds of RA fibers have a right-skewed distribution because of the anatomical constraints present in both species' skin. Meissner corpuscles, which are the sensory end-organs of RA fibers, are likely to be found deeper in the skin within dermal papilla, therefore, the thresholds can be elevated. However, the thresholds are bounded at lower end, probably due to the epidermal junction that acts as a superficial mechanical barrier for these corpuscles.     

Morphology of umbilical vessels in human fetuses: a quantitative light microscope study

OBJECTIVE: To investigate the development of the intra-abdominal part of the umbilical vessels in human fetuses by light microscopy. MATERIALS AND METHODS: The location of the umbilicus and umbilical vessels in the abdominal cavity of 90 human fetuses of gestational ages 10-40 weeks was determined. The external vessel diameter, lumen diameter, wall thickness, tunica adventitia thickness, tunica media thickness and the number of vasa vasorum were recorded from cross-sectlons of the intra-abdominal part of the umbilical vessels. 1985). 1985). RESULTS: Umbilical artery agenesis was observed on the left side in two cases and on the right in one case. There was a positive correlation between gestational age and umbilical vessel measurements. There were differences between the vessel and lumen diameters, tunica media thicknesses of the vessels of the second and third trimesters, and the full-term period. There were also predictable differences between the vessel and lumen diameters, tunica media and tunica adventitia thicknesses of the umbilical vein and umbilical arteries. CONCLUSION: Detailed information on quantitative parameters of umbilical vessels at each gestational age may prove helpful in determining pathologies of umbilical vessels and illuminating certain syndromes.     

Evaluation of false transrectal ultrasonographic pregnancy diagnoses in sheep by measuring the plasma level of pregnancy-associated glycoproteins

The present study was undertaken to investigate to what extent pregnancy diagnoses made by transrectal ultrasonography could be confirmed by measurements of plasma concentration of ovine pregnancy-associated glycoproteins (ovPAG). A total of 424 Awassi x Merino ewes were synchronized for estrus and examined by transrectal ultrasonography. In Experiment 1, the ewes (n = 156) were repeatedly scanned in a standing position on d 29, 36 and 50 of gestation. Similarly, the ewes (n = 268) in Experiment 2 were scanned on d 24, 29 and 34 of gestation, but these ewes were fasted for 12 h prior to the examination and the abdominal wall of each animal was lifted up by the hands of the assistant during the scanning. Blood samples were withdrawn after each transrectal ultrasonographic examination in both experiments. Ovine PAG concentrations were measured in plasma by a heterologous radioimmunoassay and the cut-off value for pregnancy was > or = 1 ng.mL-1. Based on the lambing performance, in Experiment 1, altogether 47 false negative and 38 false positive diagnoses were made by transrectal ultrasonography in 24 and 33 ewes, respectively between d 29 and 50 of gestation. In Experiment 2, altogether 8 false negative and 13 false positive diagnoses both were made in 7 ewes between d 24 and 34 of gestation. In both experiments, all ewes with false negative diagnoses had ovPAG concentrations higher than the threshold level for pregnancy diagnosis and all ewes with false positive diagnoses had ovPAG concentrations lower than the threshold of pregnancy. Furthermore, by the PAG-RIA test all lambed or aborted ewes (n = 63) were correctly diagnosed as pregnant and with three exceptions, all non-lambed ewes (n = 361) were correctly diagnosed as non-pregnant during the examined periods of both experiments.     

End-to-end linkage (EEL) clustering algorithm: a study on the distribution of Meissner corpuscles in the skin

A novel hierarchical clustering algorithm was applied to the distribution of Meissner corpuscles in the skin of mammals. This method, called end-to-end linkage (EEL), is useful for grouping data that consists of chain-like contingencies in the multivariable space. Unlike the traditional techniques which uncover hyperspherical clusters (e.g. single linkage), EEL considers the shortest distance between the predefined end pairs of the two clusters as an inter-group distance. This scheme allows characterizing the internal structure of data better than other hierarchical techniques. The anatomical data used in the case study is important for studying the sense of touch. The results show a substantial improvement over the traditional single-linkage method. On average, the number of correctly classified corpuscles is increased to twice the number identified by the single-linkage method. EEL can also be used for analyzing other sensory modalities where geometric relationships need to be explored. In addition, the report contains corpuscle density and epidermal-ridge width data obtained from several species.