Biomedical subjects

Parminder Kaur

Publications and source records attributed to Parminder Kaur.

14 recordsLinked to original sources

PubMed2006

Effect of diethyl maleate induced oxidative stress on male reproductive activity in mice: redox active enzymes and transcription factors expression.

Free radicals and reactive oxygen species (ROS) associated with oxidative stress are likely to play a number of significant roles in male reproduction. Present study was carried out to evaluate the effect of diethyl maleate (DEM) induced oxidative stress on male fertility in mice. Intraperitoneal injection of DEM for two weeks resulted in decrease in reduced glutathione and increase in the oxidized glutathione levels in the testis. Effect on the reproductive ability in term of sperm concentration, motility and fertility status was studied. Sperm concentration and motility were found to be significantly reduced along with a significant reduction in the litter size. Expression of redox sensitive transcription factor, cjun and cfos genes, along with gamma-glutamyl cysteine synthetase (GCS) and manganese superoxide dismutase (MnSOD) expression were also studied using RT-PCR after DEM treatment. RT-PCR analysis revealed decrease in the testicular mRNA expression for cjun and cfos whereas the expression for GCS and MnSOD increased. Enzyme activity of SOD also increased. The study reflects the effect of DEM induced oxidative stress on the reproductive ability of male mice and interplay of the various components of the antioxidant defense system and redox regulated transcription factors at the transcriptional level.

Animals↗

PubMed2006

Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry.

P21ras, the translation product of the most commonly mutated oncogene, is a small guanine nucleotide exchange protein. Oxidant-induced post-translational modifications of p21ras including S-nitrosation and S-glutathiolation have been demonstrated to modulate its activity. Structural characterization of this protein is critical to further understanding of the biological functions of p21ras. In this study, high-resolution and high mass accuracy Fourier transform mass spectrometry was utilized to map, in detail, the post-translational modifications of p21ras (H-ras) exposed to oxidants by combining bottom-up and top-down techniques. For peroxynitrite-treated p21ras, five oxidized methionines, five nitrated tyrosines, and at least two oxidized cysteines (including C118) were identified by "bottom-up" analysis, and the major oxidative modification of C118, Cys118-SO3H, was confirmed by several tandem mass spectrometry experiments. Additionally, "top-down" analysis was conducted on p21ras S-glutathiolated by oxidized glutathione and identified C118 as the major site of glutathiolation among the four surface cysteines. The present study provides a paradigm for an effective and efficient method not only for mapping post-translational modifications of proteins but also for predicting the relative selectivity and specificity of oxidative post-translational modifications, especially using top-down analysis.

Amino Acid Sequence↗

PubMed2006

Novel approach to multifunctionalized homoallylic alcohols via regioselective ring opening of aryl oxiranes with 3-iodo allenoates.

[reaction: see text] 3-Iodo allenoates were generated in situ and utilized, for the first time, in the ring opening of oxiranes in a regioselective fashion. This simple one-pot three-component reaction protocol provides easy access to highly functionalized homoallylic alcohols in good yields and moderate to very good (Z/E) selectivity. The two functional groups (ester and halogen) can be further subjected to many synthetic transformations.

Alcohols↗

PubMed2006

Tertiary-butyl hydroperoxide induced oxidative stress and male reproductive activity in mice: role of transcription factor NF-kappaB and testicular antioxidant enzymes.

Reactive oxygen species (ROS) have been proposed as a major factor affecting male reproductive capacity. The present study has evaluated the possible role of oxidative stress during testicular pathogenesis in male mice exposed to tertiary-butyl hydroperoxide (TBHP). TBHP was administered by daily intraperitoneal injection for 2 weeks. Treatment greatly increased lipid peroxidation in the testis and led to a significant decrease in sperm concentration and motility and a reduction in litter size relative to controls. An increase in testicular mRNA abundance of redox-regulated p50 and p65 subunits of NF-kappaB was observed after TBHP treatment. Evaluation of NF-kappaB regulated antioxidant enzymes in the testis revealed an increase in superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione-S-transferase (GST) activities and corresponding mRNA abundance. These results suggest a potential role of NF-kappaB in oxidative stress mediated changes in the physiology of male reproductive system.

Animals↗

PubMed2006

Algorithms for automatic interpretation of high resolution mass spectra.

Automated interpretation of high-resolution mass spectra in a reliable and efficient manner represents a highly challenging computational problem. This work aims at developing methods for reducing a high-resolution mass spectrum into its monoisotopic peak list, and automatically assigning observed masses to known fragment ion masses if the protein sequence is available. The methods are compiled into a suite of data reduction algorithms which is called MasSPIKE (Mass Spectrum Interpretation and Kernel Extraction). MasSPIKE includes modules for modeling noise across the spectrum, isotopic cluster identification, charge state determination, separation of overlapping isotopic distributions, picking isotopic peaks, aligning experimental and theoretical isotopic distributions for estimating a monoisotopic peak's location, generating the monoisotopic mass list, and assigning the observed monoisotopic masses to possible protein fragments. The method is tested against a complex top-down spectrum of bovine carbonic anhydrase. Results of each of the individual modules are compared with previously published work.

Algorithms↗

PubMed2005

Characterization of a new qQq-FTICR mass spectrometer for post-translational modification analysis and top-down tandem mass spectrometry of whole proteins.

The use of a new electrospray qQq Fourier transform ion cyclotron mass spectrometer (qQq-FTICR MS) instrument for biologic applications is described. This qQq-FTICR mass spectrometer was designed for the study of post-translationally modified proteins and for top-down analysis of biologically relevant protein samples. The utility of the instrument for the analysis of phosphorylation, a common and important post-translational modification, was investigated. Phosphorylation was chosen as an example because it is ubiquitous and challenging to analyze. In addition, the use of the instrument for top-down sequencing of proteins was explored since this instrument offers particular advantages to this approach. Top-down sequencing was performed on different proteins, including commercially available proteins and biologically derived samples such as the human E2 ubiquitin conjugating enzyme, UbCH10. A good sequence tag was obtained for the human UbCH10, allowing the unambiguous identification of the protein. The instrument was built with a commercially produced front end: a focusing rf-only quadrupole (Q0), followed by a resolving quadrupole (Q1), and a LINAC quadrupole collision cell (Q2), in combination with an FTICR mass analyzer. It has utility in the analysis of samples found in substoichiometric concentrations, as ions can be isolated in the mass resolving Q1 and accumulated in Q2 before analysis in the ICR cell. The speed and efficacy of the Q2 cooling and fragmentation was demonstrated on an LCMS-compatible time scale, and detection limits for phosphopeptides in the 10 amol/muL range (pM) were demonstrated. The instrument was designed to make several fragmentation methods available, including nozzle-skimmer fragmentation, Q2 collisionally activated dissociation (Q2 CAD), multipole storage assisted dissociation (MSAD), electron capture dissociation (ECD), infrared multiphoton induced dissociation (IRMPD), and sustained off resonance irradiation (SORI) CAD, thus allowing a variety of MS(n) experiments. A particularly useful aspect of the system was the use of Q1 to isolate ions from complex mixtures with narrow windows of isolation less than 1 m/z. These features enable top-down protein analysis experiments as well structural characterization of minor components of complex mixtures.

Equipment Design↗

PubMed2005

Effect of selenium-induced oxidative stress on the cell kinetics in testis and reproductive ability of male mice.

OBJECTIVE: The present study evaluated the role of experimental oxidative stress (induced by feeding diets with different concentrations of selenium [Se], a trace nutrient and potent antioxidant) on male reproductive activity in mice. METHODS: To create different levels of oxidative stress in male mice, three diets with different levels of Se were fed to different groups for 8 wk. Mice in group 1 were fed a yeast-based diet, which is considered a Se-deficient diet (0.02 ppm). Mice in groups 2 and 3 were fed with an Se-deficient diet supplemented with 0.2 and 1 ppm Se as sodium selenite, respectively. RESULTS: After completion of the feeding schedule, a significant decrease in Se levels were observed in Se-deficient mice (group 1), whereas Se levels greatly increased in the Se-excess mice (group 3). Glutathione peroxidase activity was greatly decreased in the liver and testis in group 1, whereas glutathione-S-transferase activity was significantly increased in the testis. No significant change was found in activities of glutathione peroxidase and glutathione-S-transferase in group 3 compared with group 2. Cell kinetics showed a significant decrease in the number of pachytene spermatocytes and young and mature spermatids in group 1 compared with group 2. No appreciable change was observed in the germinal cell population in group 3. A significant decrease in sperm number was observed in group 1 compared with group 2. No change in these parameters was observed in group 3. The fertility status of mice in terms of percent fertility and litter size also exhibited a significant decrease in the reproductive ability of group 1. No change in these parameters was observed in group 3 compared with group 2. CONCLUSION: The present results clearly demonstrate the effect of oxidative stress generated by feeding different concentrations of Se on cell kinetics in the testis and, hence, its effect on the reproductive ability of male mice.

Animals↗

PubMed2005

Selenium, a versatile trace element: current research implications.

Selenium (Se), a trace element, has evolved from its toxic properties to an essential element. Se was known a potent antioxidant through glutathione peroxidase (selenium being part of this molecule). Later, many other selenium-binding proteins were discovered and their functions were tried to be known with unsuccessful results in many cases. Se is known to be involved in carcinogenesis, immune function, male reproduction, cardiovascular diseases etc. The specific mechanism of the involvement of the element is still not known. Recent research with application of modern research tools viz. bioinformatics, cDNA microarray and transgenesis have revealed the mechanism of selenium involvement in various processes. This review highlights mysterious and useful roles of selenium in biological processes.

Animals↗

PubMed2004

Use of statistical methods for estimation of total number of charges in a mass spectrometry experiment.

Estimation of the number of ions in a mass spectrometry experiment is needed to determine instrumentation parameters such as ionization efficiency, collision-induced dissociation efficiency, ion-transfer efficiency, ion trapping efficiency, and preamplifier detection limit. This work aims at analyzing the statistical characteristics (primarily variance) in the intensities of the isotopic distributions, which depend on the number of ions in the cell. A mathematical derivation was developed based on the maximum likelihood estimation method, which estimates the most likely number of ions in the cell using a method known as nonrandom parameter estimation. The performance of the method improves with increase in the number of observed distributions. The method works well provided the spectra show isotopic resolution and is independent of the instrument or method used to arrive at the spectra.

Carbon Isotopes↗

PubMed2004

A high pressure matrix-assisted laser desorption ion source for Fourier transform mass spectrometry designed to accommodate large targets with diverse surfaces.

A new design for a high pressure MALDI-FTMS instrument is described and initial data are shown. The instrument incorporates a large, 10 cm x 10 cm, sample translation stage to accommodate and position the MALDI target. The new instrument allows coupling to a wide variety of surface techniques such as gel electrophoresis or surface plasmon resonance. Coupling to thin layer chromatography is shown. Furthermore, a new nozzle design allows high pressure collisional cooling sufficient to stabilize gangliosides while minimizing the gas load on the system.

Chromatography, Thin Layer↗

PubMed2004

Effect of selenium-induced oxidative stress on the oxidation reduction system and reproductive ability of male mice.

The present study was carried out to evaluate the effect of selenium (Se)-induced oxidative stress on the oxidation reduction system and the fertility status of male mice. Different levels of Se, a potent antioxidant, were fed in three separate groups for 8 wk to create the different oxidative stress in mice. A significant decrease in the glutathione peroxidase (GSH-Px) in both liver and testis was observed in the Se-deficient (0.02 ppm) group I, whereas enzyme levels in the Se-excess (1 ppm) group were comparable to the Se-adequate (0.2 ppm) group. Glutathione-S-transferase activity was enhanced in group I in comparison to group II; however, no change was seen in group III. The glutathione reductase and superoxide dismutase activities were decreased in the Se-deficient group, whereas the enzyme levels were significantly increased in the Se-excess group. The fertility status of the animals studied in terms of percentage fertility and litter size showed a significant decrease in the reproductive ability of male mice in group I when compared to group II. No changes in the fertility status of animals were observed in group III. Thus, the data clearly indicate the effect of oxidative stress generated by feeding various Se levels on the oxidation reduction system and, consequently, its effect on the reproductive ability of male mice.

Animals↗

PubMed2004

Influence of selenium induced oxidative stress on spermatogenesis and lactate dehydrogenase-X in mice testis.

AIM: To evaluate the effect of oxidative stress on the spermatogenesis and lactate dehydrogenase-X (LDH-X) activity in mouse testis. METHODS: For creating different levels of oxidative stress in mice, three selenium (Se) level diets were fed in separate groups for 8 weeks. Group 1 animals were fed yeast-based Se-deficient (0.02 ppm) diet. Group 2 and Group 3 animals were fed with the same diet supplemented with 0.2 ppm and 1 ppm Se as sodium selenite, respectively. After 8 weeks, biochemical and histopathological observations of the testis were carried out. LDH-X levels in the testis were analyzed by western immunoblot and ELISA. RESULTS: A significant decrease in testis Se level was observed in Group 1 animals, whereas it was enhanced in Group 3 as compared to Group 2. The glutathione peroxidase (GSH-Px) activity was significantly reduced in both the liver and testis in Group 1, but not in Group 2 and 3. A significant increase in the testis glutathione-S-transferase (GST) activity was observed in Group 1, whereas no significant change was seen in Groups 2 and 3. Histological analysis of testis revealed a normal structure in Group 2. A significant decrease in the germ cell population in Group 1 was observed as compared to Group 2 with the spermatids and mature sperm affected the most. Decrease in the lumen size was also observed. In the Se-excess group (Group 3), displacement of germ cell population was observed. Further, a decrease in the LDH-X level in testis was observed in Group 1. CONCLUSION: Excessive oxidative stress in the Se deficient group, as indicated by changes in the GSH-Px/GST activity, affects the spermatogenic process with a reduction in mature sperm and in turn the LDH-X level.

Animals↗

PubMed2003

Carcinogenicity of nickel is the result of its binding to RNA and not to DNA.

In vivo and in vitro studies have shown for the first time that nickel binds maximally to RNA as compared to DNA. The distribution of radiolabeled nickel in various subcellular fractions of liver, kidney, spleen, and lungs of rats has shown that it was mostly localized in the nuclear fraction of each organ, where it was bound to RNA and not to DNA or to proteins. Significant differences between infrared (IR) spectra of RNA and RNA incubated with nickel chloride further corroborate these findings. Parallel experiments with DNA did not show any such effects. Nickel may, therefore, be acting as a carcinogen by controlling gene expression post-transcriptionally.

Animals↗

PubMedDate not supplied

Effect of experimental oxidative stress on steroidogenesis and DNA damage in mouse testis.

The objective of the present study was to evaluate the effect of oxidative stress induced by feeding various levels of selenium on steroidogenesis and DNA damage in mouse testes. To create various levels of oxidative stress in mice, diets with three different Se levels were fed to separate groups for 8 weeks. Group 1 animals were fed a yeast-based diet, which was considered a Se-deficient diet (0.02 ppm). Group 2 and 3 animals were fed a Se-deficient diet supplemented with 0.2 and 1 ppm Se as sodium selenite, respectively. After completion of the diet feeding, estimations were carried out, and results were compared with those of group 2. A significant decrease in Se levels was observed in group 1 animals, whereas they were greatly enhanced in group 3. Glutathione peroxidase (GSH-Px) activity was greatly reduced in both the liver and testes in group 1, whereas no significant changes were found in GSH-Px activity in group 3. Serum luteinizing hormone, follicle-stimulating hormone (FSH), and testosterone levels were reduced in group 1. Significant decreases of sperm number and motility were observed in group 1 when compared to group 2 male mice. No changes in these parameters were observed in group 3. DNA fragmentation was observed in both groups 1 and 3; however, the damage was more prevalent in group 1. The results clearly demonstrate the effect of oxidative stress generated by feeding various Se levels on the steroidogenesis and DNA fragmentation in mice testes.

Animals↗