The aim of the experiment was to determine the effect of potential antioxidants (adenosine, L-cysteine hydrochloride, ascorbic acid, magnesium fumarate and prolactin) supplementing the Biosolwens ...extender on semen survival time and sperm chromatin structure. The semen motility was examined every day and the susceptibility of sperm chromatin to denaturation was evaluated on collection day and day 15 of storage. The addition of magnesium fumarate to Biosolwens extender increased sperm survival but resulted in the highest increment in the proportion of sperm with damaged chromatin. Biosolwens supplemented with 200 mg of L-cysteine hydrochloride brought the best results. It is possible that lower concentrations of this component would act in a more protective manner. The examination of the chromatin structure appears to be an useful tool for investigation of semen preservation.
We have determined the physical and genetic map of the 73,000 base-pair mitochondrial genome of a novel yeast species Saccharomyces douglasii. Most of the protein and RNA-coding genes known to be ...present in the mitochondrial DNA of Saccharomyces cerevisiae have been identified and located on the S. douglasii mitochondrial genome. The nuclear genomes of the two species are thought to have diverges some 50 to 80 million years ago and their nucleo-mitochondrial hybrids are viable but respiratorily deficient. The mitochondrial genome of S. douglasii displays many interesting features in comparison with that of S. cerevisiae. The three mosaic genes present in both genomes are quite different with regard to their structure. The S. douglasii COXI gene has two new introns and is missing the five introns of the S. cerevisiae gene. The S. douglasii cytochrome b gene has one new intron and lacks two introns of the S. cerevisiae gene. Finally, the L-rRNA gene of S. douglasii, like that of S. cerevisiae, has one intron of which the structure is different. Another salient feature of the S. douglasii mitochondrial genome reported here is that the gene order is different in comparison with S. cerevisiae mitochondrial DNA. In particular, a segment of approximately 15,000 base-pairs including the genes coding for COXII and S-rRNA has been translocated to a position between the genes coding for var1 and L-rRNA.
A semidominant nuclear suppressor, called nam6, of oxi2-V276 mitochondrial mutation has been isolated and characterized. The nuclear character of nam6 was proved by its retention in rho degree ...strains, lack of mitotic segregation in diploids and meiotic 2:2 segregation in tetrads. The specificity of nam6 was tested on 315 mit- mutations of four mitochondrial genes (oxi1, oxi2, oxi3, and cob-box). It suppresses clearly only three mutations in the oxi2 gene, restoring partially or completely cytochrome aa3 formation. The results suggest a functional character of the suppression.
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•New arylpiperazine oxicams derivatives were designed and synthesized.•Compounds obtained proved to be between two and five times analgesically more potent than the reference drug ...piroxicam.•New derivatives did not show any ulcerogenic activity.
Gastrotoxicity continues to be a major issue in therapy with nonsteroidal anti-inflammatory drugs (NSAIDs). Medicine is yet to develop absolutely safe analgesics. Numerous strategies are employed to discover new, safer NSAIDs, for example selective inhibition of cyclooxygenase-2, new molecular targets (e.g. microsomal prostaglandin E2 synthase-1), incorporation of cytoprotective compounds in the drug molecule or modification of the classic NSAIDs currently available on the market. The research presented in this paper is indicative of a current worldwide trend in this area of science, and is an example of the fourth strategy noted above. Two series of new arylpiperazine derivatives of the classic NSAID – piroxicam, were developed by conventional synthesis. The full range of compounds obtained proved to be between two and five times analgesically more potent than the reference drug and, most importantly, they did not show any ulcerogenic activity.
Introduction Antioxidant enzymes protect the human body against the harmful effects of oxidative stress. The activity of antioxidant enzymes changes with age and depends on dietary nutrients such as ...fats and vitamins, which can have a significant impact on minimizing or exacerbating oxidative stress. Aim To examine the effect of age, BMI, diet, physical activity, and smoking status on the activity of erythrocyte antioxidant enzymes catalase, glutathione reductase, glutathione peroxidase glutathione S-transferase, superoxide dismutase, and glutathione concentrations in healthy women. Material and methods This study included 98 healthy women aged between 20 and 65 years. All women underwent anthropometric tests: body weight, height, hip, and waist circumference. Antioxidant activity in erythrocytes was measured by spectrophotometric methods. Results Catalase activity increased significantly with age (p < 0.001), while superoxide dismutase activities and glutathione decreased with age (p = 0.008, p = 0.023, respectively). Women with a lower BMI (emaciation) had higher superoxide dismutase activity than those in the first degree of obesity (p = 0.009). Conclusions (1) Increased catalase activity with age may signify a large amount of hydrogen peroxide resulting from malfunctioning antioxidant systems in old age. (2) A decline in superoxide dismutase activity with age may indicate inactivation of this enzyme, inappropriate SOD function in the presence of excessive amounts of hydrogen peroxide, and glycation of superoxide dismutase molecules. (3) A negative correlation between superoxide dismutase activity and the BMI index may indicate a decreased enzymatic activity in obese people. Keywords: Oxidative stress, Antioxidant enzymes, Obesity, Physical activity, Diet
ATP level, cell motility and viability, oxygen uptake, pyruvate kinase activity, and ultra-weak photon emission (UPE) induced by red-ox Fe(2+)-ascorbate cycling system were studied in fresh, in ...previously equilibrated in a glycerol diluent, and in cryopreserved bull spermatozoa, exposed to thermal stress by incubation of the cells at 44 degrees C. A sharp drop in motility and viability of fresh spermatozoa and even more so, of equilibrated and cryopreserved cells was accompanied by accumulation of ATP. When cell movement was totally inhibited, ATP utilization was decreased, while chemical energy continued to be produced by cell pyruvate kinase, one of the key glycolytic enzymes, which in spermatozoa is very active (6500 IU/g protein) and insensitive to feed-back inhibition by excess of ATP and L-cysteine. Accumulation of ATP during incubation at 44 degrees C in 0.9% NaCl was accompanied by rapid decrease in oxygen consumption by fresh spermatozoa and an increase in Fe(2+)-ascorbate induced UPE, followed by a sharp decrease in ATP level observed at the end of induced UPE measurement. The increase in photon emission due to lipid peroxidation was highly correlated with the increase in cell ATP level caused by thermal stress.
Janus green B was found to be a specific inhibitor of mitochondrial function in yeast. This is consistent with the Janus green specificity in supravital staining of mitochondria. A mutant of S. ...Cerevisiae resistant to Janus green B was isolated. It shows cross resistance to oligomycin, ethidium bromide and a weak resistance to chlormaphenicol. The mutant was found to be sensitive to cycloheximide and erythromycin. Genetic analysis of this mutant showed that mitochondrial genes are not involved in the determination of Janus Green resistance. Tetrad analysis suggested that two more more nuclear genes are concerned, but many unusal genetic features suggestive of the involvement of a cytoplasmic element remain to be explained.
We report the genetic characterization, molecular cloning, and sequencing of a novel nuclear suppressor, the NAM9 gene from Saccharomyces cerevisiae, which acts on mutations of mitochondrial DNA. The ...strain NAM9-1 was isolated as a respiration-competent revertant of a mitochondrial mit mutant which carries the V25 ochre mutation in the oxil gene. Genetic characterization of the NAM9-1 mutation has shown that it is a nuclear dominant omnipotent suppressor alleviating several mutations in all four mitochondrial genes tested and has suggested its informational, and probably ribosomal, character. The NAM9 gene was cloned by transformation of the recipient oxil-N25 mutant to respiration competence by using a gene bank from the NAM9-1 rho° strain. Orthogonal-field alternation gel electrophoresis analysis and genetic mapping localized the NAM9 gene on the right arm of chromosome XIV. Sequence analysis of the NAM9 gene showed that it encodes a basic protein of 485 amino acids with a presequence that could target the protein to the mitochondrial matrix. The N-terminal sequence of 200 amino acids of the deduced NAM9 product strongly resembles the S4 ribosomal proteins from chloroplasts and bacteria. Significant although less extensive similarity was found with ribosomal cytoplasmic proteins from lower eucaryotes, including S.
cerevisiae. Chromosomal inactivation of the NAM9
+
gene is not lethal to the cell but leads to respiration deficiency and loss of mitochondrial DNA integrity. We conclude that the NAM9 gene product is a mitochondrial ribosomal counterpart of S4 ribosomal proteins found in other systems and that the suppressor acts through decreasing the fidelity of translation.
To understand better the factors contributing to keratoconus (KTCN), we performed comprehensive transcriptome profiling of human KTCN corneas for the first time using an RNA-Seq approach. Twenty-five ...KTCN and 25 non-KTCN corneas were enrolled in this study. After RNA extraction, total RNA libraries were prepared and sequenced. The discovery RNA-Seq analysis (in eight KTCN and eight non-KTCN corneas) was conducted first, after which the replication RNA-Seq experiment was performed on a second set of samples (17 KTCN and 17 non-KTCN corneas). Over 82% of the genes and almost 75% of the transcripts detected as differentially expressed in KTCN and non-KTCN corneas were confirmed in the replication study using another set of samples. We used these differentially expressed genes to generate a network of KTCN-deregulated genes. We found an extensive disruption of collagen synthesis and maturation pathways, as well as downregulation of the core elements of the TGF-β, Hippo, and Wnt signaling pathways influencing corneal organization. This first comprehensive transcriptome profiling of human KTCN corneas points further to a complex etiology of KTCN.
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