Similar Quantitative Changes Research Articles

Neuroprotective effects of isatin and afobazole in rats with rotenone-induced Parkinsonism are accompanied by increased brain levels of Triton X-100 soluble alpha-synuclein.

Effects of the endogenous neuroprotector isatin and the pharmacological drug afobazole (exhibiting neuroprotective properties) on behavioral reactions and quantitative changes in the brain proteomic profile have been investigated in rats with experimental rotenone Parkinsonism. A single dose of isatin (100 mg/kg subcutaneously on the last day of a 7-day course of rotenone administration) improved the motor activity of rats with rotenone-induced Parkinsonism in the open field test (horizontal movements) and the rotating rod test. Afobazole (10 mg/kg intraperitoneally, daily during the 7-day course of rotenone administration) reduced the manifestations of rigidity and postural instability. Proteomic analysis, performed using brain samples obtained the day after the last administration of rotenone and neuroprotectors, revealed similar quantitative changes in the brain of rats with rotenone Parkinsonism. An increase in the relative content of 65 proteins and a decrease in the relative content of 21 proteins were detected. The most pronounced changes - an almost ninety-fold increase in the alpha-synuclein content - were found in the brains of rats treated with isatin. In animals of the experimental groups treated with "Rotenone + Isatin", as well as "Rotenone + Afobazole", the increase in the relative content of this protein in the brain was almost 60 and 50 times higher than the control values. Taking into consideration the known data on the physiological role of alpha-synuclein, an increase in the content of this protein in the brain upon administration of neuroprotectors to animals with rotenone Parkinsonism may represent a compensatory reaction, at least in the early stages of this disease and the beginning of its treatment.

Pregnancy preparation: redistribution of CCR7-positive cells in the rat uterus.

Changes in the endometrium prior to implantation may be critical in predicting pregnancy outcomes. This study shows that the endocrine system directs positional changes in CCR7+ cells before implantation, which may be critical for developing maternal tolerance. Suppression of the maternal immune system is vital for the implantation of the semi-allogeneic embryo. Although progress in understanding the dialogue between mother and embryo has been made, key interactions between maternal immune cells, hormones, and chemokines remain elusive. Uterine expression of the C-C chemokine receptor type 7 (CCR7) could recruit T regulatory cells and facilitate localized immune suppression. To test this concept, Ccr7 mRNA and protein were assessed in uterine tissue. Ccr7 mRNA expression peaked at day 4 in pregnant rat uteri and then declined at days 5 and 6. CCR7 protein showed similar quantitative changes. To test if female sex steroids affected the spatial distribution of CCR7-expressing cells, uteri from ovariectomized rats, progesterone-pretreated rats (2 mg daily), and progesterone-pretreated rats injected with estradiol (0.2 µg) were analyzed. Progesterone increased CCR7-positive (+) cells in the antimesometrial stroma. Progesterone and estradiol increased CCR7+ cells in the mesometrial stroma. Estradiol increased the density of cluster of differentiation 4 (CD4) positive cells in the mesometrial stromal region over progesterone alone. The density of cells expressing the T regulatory cell marker, forkhead box protein 3 (FOXP3), increased in the antimesometrial stroma in response to progesterone alone. Progesterone and estradiol increased FOXP3+ cells in the antimesometrial region of the stroma. Co-localization of CCR7, CD4, and FOXP3 in the stroma suggests CCR7+ cells are T regulatory cells. Polarization of CCR7+ cells in the endometrial stroma was an intrinsic response regulated by sex steroids and did not require the presence of an embryo.

Profiling of Host Cell Response to Successive Canine Parvovirus Infection Based on Kinetic Proteomic Change Identification.

Canine parvovirus (CPV) reproduces by co-opting the resources of host cells, inevitably causing cytotoxic effects to the host cells. Feline kidney F81 cells are sensitive to CPV infection and show disparate growing statuses at different time points post-infection. This study analysed the response of F81 cells to CPV infection at successive infection time points by iTRAQ-based quantitative proteomics. Differentially expressed proteins (DEPs) during 60 h of infection and at selected time points post-infection were identified by an analysis of variance test and a two-tailed unpaired t test, respectively. DEPs with similar quantitative changes were clustered by hierarchical clustering and analysed by gene ontology enrichment, revealing that 12 h and 60 h post-infection were the optimal times to analyse the autonomous parvovirus replication and apoptosis processes, respectively. Using the MetacoreTM database, 29 DEPs were enriched in a network involved in p53 regulation. Besides, a significantly enriched pathway suggests that the CPV-induced cytopathic effect was probably due to the deficiency of functional CFTR caused by CPV infection. This study uncovered the systemic changes in key cellular factors involved in CPV infection and help to understand the molecular mechanisms of the anti-cancer activity of CPV and the cytopathic effects induced by CPV infection.

Inductively coupled mass spectrometry analysis of biometals in conditional Hamp1 and Hamp1 and Hamp2 transgenic mouse models

Hepcidin, a circulatory antimicrobial peptide, is involved in iron homeostasis, inflammation, infection and metabolic signals. Humans express one hepcidin gene, HAMP but mice express two hepcidin genes, Hamp1 and Hamp2. Consecutive gene targeting events were performed to produce transgenic mice expressing conditional alleles of either Hamp1 or both Hamp1 and Hamp2 (Hamp1/2). The deletion of Hamp1 alleles elevated Hamp2 expression, particularly in males, which was reduced by endotoxin treatment. The tissue levels of iron and other biometals were quantified by inductively coupled mass spectrometry. The ubiquitous or liver-specific deletion of Hamp1 alleles yielded similar quantitative changes in iron levels in the liver, duodenum, spleen, kidney, heart and brain. The introduction of Hamp2 null allele did not exacerbate the iron-related phenotype of Hamp1 null allele. Besides iron, Hamp1 null allele significantly elevated the levels of selenium in the liver, manganese in the liver and duodenum, and copper in the brain. Mice with conditional Hamp alleles will be useful to determine the tissue-specific regulation and functions of Hamp1 and Hamp2 in biometal homeostasis and other biological processes.

Cardiorespiratory performance and blood chemistry during swimming and recovery in three populations of elite swimmers: Adult sockeye salmon

Every year, millions of adult sockeye salmon (Oncorhynchus nerka) perform an arduous, once-in-a-lifetime migration up the Fraser River (BC, Canada) to return to their natal stream to spawn. The changes in heart rate, stroke volume, and arterio-venous oxygen extraction (i.e., factors determining rates of oxygen delivery to the tissues by the cardiovascular system) have never been directly and simultaneously measured along with whole animal oxygen uptake in a maximally swimming fish. Here, such measurements were made using three sockeye salmon populations (Early Stuart, Chilko and Quesnel), which each performed two consecutive critical swimming speed (Ucrit) challenges to provide a comprehensive quantification of cardiovascular physiology, oxygen status and blood chemistry associated with swimming and recovery. Swim performance, oxygen uptake, cardiac output, heart rate and stroke volume did not significantly vary at rest, during swimming or during recovery between populations or sexes. Despite incomplete metabolic recovery between swim challenges, all fish repeated their swim performance and similar quantitative changes in the cardiorespiratory variables were observed for each swim challenge. The high maximum cardiorespiratory performance and excellent repeat swim performance are clearly beneficial in allowing the salmon to maintain steady ground speeds and reach the distant spawning grounds in a timely manner.

Quantitative proteome analysis of cisplatin‐induced apoptotic Jurkat T cells by stable isotope labeling with amino acids in cell culture, SDS‐PAGE, and LC‐MALDI‐TOF/TOF MS

Quantitative proteome analysis of cisplatin-induced apoptosis in total Jurkat T cell lysates was performed in order to identify modified proteins. Proteins were labeled in cell culture with stable isotopes of arginines, and fractionated by SDS-PAGE. Subsequently, tryptic peptides were analyzed by nano-LC coupled offline to MALDI-TOF/TOF-MS as an alternative to commonly used online LC-ESI-MS. As a result, 26 proteins were found with a relative abundance higher than 1.5, thereof 19 already known and seven unknown to be involved in apoptosis (adenine phosphoribosyltransferase, microsomal signal peptidase 25 kDa subunit, phosphomevalonate kinase, probable rRNA processing protein EBP2, RNA-binding protein 4, transmembrane protein 33, and tetratricopeptide repeat domain 9C). Immunoblotting of core-binding factor beta and elongation factor 2 revealed similar quantitative changes as detected by the SILAC-based proteomics approach. Strikingly, 8 of 26 identified apoptosis-modified proteins contained at least one RNA-binding motif. Three caspase cleavage sites of the 54 kDa nuclear RNA-binding protein (p54nrb) were mapped at DQLD(231) (downward arrow)D, DQVD(286) (downward arrow)R, and MMPD(422) (downward arrow)G by applying caspase-3 to the in vitro translated protein and mutation analysis. The determined caspase cleavage sites were located C-terminal to the two RNA-binding motifs and one (DQLD(231) (downward arrow)D) within the NOPS domain of p54nrb. Concisely, quantitative protein data generated by offline LC-MALDI-MS were shown to be particularly accurate. Furthermore, only regulated peptides were selected in a result-dependent manner for MS/MS analyses and revealed novel apoptosis-modified proteins.

Morphometric and Ultrastructural Changes in Ciliated Cells of the Oviductal Epithelium in Prolific Chinese Meishan and Large White Pigs during the Oestrous Cycle

There are some differences in reproductive features between the Chinese Meishan (CM) pig and Large White (LW) pig. The aim of the present study was to investigate the quantitative changes and ultrastructural features of ciliated cells in the various regions of the CM and LW pig oviduct during the follicular and luteal phases of the oestrous cycle. In the fimbrial and ampullar epithelia at the follicular phase, the ciliated cells were more plentiful than in the isthmic region in both pigs. In the CM pigs, there was a striking decrease in the percentage and cell height of ciliated cells in the luteal phase compared with the follicular phase. Although similar quantitative changes were observed in the LW pig oviduct, these changes were less dramatic than that in the CM pig oviduct. In both pigs, the percentage and cell height of ciliated cells in the isthmus were unchanged between the follicular and luteal phases. The ultrastructure of ciliogenic and ciliated cells was observed. In the fimbrial and ampullar epithelia during the follicular phase, most of the ciliated cells showed normal morphology, having many elongated cilia and mitochondria, but in the CM pig oviduct during the luteal phase many of the ciliated cells possessed immature cilia and swollen mitochondria. Cells undergoing ciliogenesis were frequently observed in the fimbriae and ampulla, and occasionally in the isthmus. Cytoplasmic protrusions containing variable numbers of ciliary axonemal complexes occurred in the fimbrial and ampullar epithelium in the CM pigs at the luteal phase, suggesting that deciliation of cells occurs by membrane-bound cilia packets forming at the apices of cells and pinching off. These results demonstrate that there are regional variations in the cyclic changes associated with the oviductal ciliated cells of the pigs, while there are marked morphometrical and ultrastructural changes in oviductal ciliated cells of the CM pigs compared with that of the LW pigs.

Integration of morphological and cytophysiological studies on estrogen receptor

Ultrastructural and immunocytochemical studies of an intra-nuclear particle, the perichromatin granule (PCG), demonstrated the presence of processed mRNA in this structure. Ovariectomy caused an increase in the number of PCGs in uterine cells and administration of estradiol drastically reduced the nuclear pool of PCGs in 15 min. In vitro studies demonstrated that this depletion was accompanied by an increase of the export of previously synthesized RNA. Similar quantitative changes of the abundance of PCG and of the rate of the export of RNA were found in ventral prostate after orchiectomy and testosterone restitution, as well as in the target cells of FSH, LH, TSH, and ACTH. These results taken together led us to conclude that PCGs constitute an intra-nuclear compartment of a few processed mRNA in equilibrium with transcription and export. This mRNA is rapidly transferred to the cytoplasm by specific hormone signals.

Quantitative MRI in patients with idiopathic generalized epilepsy. Evidence of widespread cerebral structural changes.

In patients with idiopathic generalized epilepsy (IGE), visual inspection of routine MRI is normal. However, pathological studies have shown microdysgenesis in grey and white matter in a large percentage of autopsies from cases of IGE. Recently, widespread structural changes not evident on visual inspection of high resolution MRI have been shown using quantitative MRI in patients with apparently focal cerebral dysgenesis. We sought to determine whether similar quantitative changes might be present in patients with IGE, reflecting possible underlying structural abnormalities. Twenty patients with juvenile myoclonic epilepsy, 10 patients each with childhood absence epilepsy and juvenile absence epilepsy, five patients with tonic-clonic seizures on awakening and 30 control subjects had T1-weighted volume acquisition MRI scans on a 1.5T GE scanner. The cerebral hemispheres were segmented semi-automatically, allowing the comparison of normalized cortical and subcortical matter volumes between groups, and investigation of the regional distribution of cortical and subcortical matter in individual subjects. Patients with IGE had significantly larger cortical grey matter volumes than control subjects. Significant abnormalities of the regional distribution of cerebral grey and subcortical matter were found in eight out of 20 patients with juvenile myoclonic epilepsy, one out of 10 patients with childhood absence epilepsy, four out of 10 patients with juvenile absence epilepsy and two out of five patients with tonic-clonic seizures on awakening, but in none of the 30 control subjects. Using MRI-segmentation, we identified widespread cerebral structural changes in patients with various IGE syndromes. Quantitative MRI supports the existence of structural abnormalities in patients with IGE.

Non-histone nuclear proteins in liver of Japanese quail divergently selected for relative susceptibility or resistance to starvation

1. 1. Nucleoplasmic, 0.35 M NaCl- and 5 M urea-soluble non-histone proteins were isolated from the liver of females of Japanese quail selected for the largest (a susceptible line) and the smallest (a resistant line) decreases in body weight at 12 weeks of age following a 4-day starvation. 2. 2. While the resistant line showed a specific increase in the amount of 40- and 60-kDa non-histones soluble in 0.35 M NaCl, the line sensitive to starvation did not exhibit significant changes in protein composition in comparison with a control line mated at random. 3. 3. After a 4-day starvation, the all genetic groups of birds developed similar quantitative changes among nucleoplasmic proteins. The 0.35 M NaCl- and 5 M urea-soluble proteins were, however, not affected by fasting.

Interrelationships Between Serologic Markers of Immune Activation and T Lymphocyte Subsets in HIV Infection

Relationships among four serologic activation markers and T cell subsets were measured in HIV-seropositive former blood donors (N = 64) and seronegative controls (N = 61). Significant correlations were observed for the HIV group in pairwise comparisons of soluble IL-2 receptor (sIL-2R), beta 2-microglobulin (beta 2M), neopterin (NEOP), and soluble CD8 (sCD8). CD4 cell levels (number/microliter) in the HIV group showed significant negative correlation with all four serologic markers; CD8 cell levels, in contrast, showed no significant correlation with any serologic activation marker measured. Significant correlations were observed, however, among various cell surface activation markers and serologic activation markers. Specifically, the proportion of CD8 cells expressing CD45RA showed significant negative correlations with NEOP and B2M levels, whereas the proportion of CD8 cells expressing HLA-DR showed significant positive correlations with B2M and sIL-2R levels. Further, the proportion of CD8 cells expressing CD38 showed significant positive correlations with all four serologic activation markers. These findings indicate that sIL-2R, B2M, NEOP, and sCD8 show similar quantitative changes and correlational relationships to CD4 cell destruction in HIV infection; they differ, however, in their relationships to proportional changes in activated CD8 cell subsets.

Responsiveness of ewe genotypes to varying nutritional management, and conventional genotype × environment interactions in sheep

ABSTRACTAn experiment was conducted during 2 years to evaluate industry Coopworth, industry Romney and Waihora Romney ewes (a strain selected for 10 years for increased ewe productivity) in a variety of nutritional and management treatment combinations. Waihora Romneys were most productive with small and inconsistent differences between industry Romneys and Coopworths. Conventional cross-classified analyses of variance did not reveal large, consistent nor economically important ewe genotype × nutritional treatment or year interactions. Additional analyses were designed to investigate whether the ewe genotypes responded to changes in the nutritional and(or) management environment with similar quantitative changes in production. A sensitive genotype would be one that responded to environmental changes with relatively large changes in productivity; a stable genotype would be one whose changes in production in response to a variable environment were smaller. By several statistical procedures designed to assess relative ‘sensitivity’ v. ‘stability’, Waihora Romney ewes appeared to be the most stable, with industry Romney and Coopworth ewes similar in their sensitivity.

Cytological responses of the pituitary (rostral pars distalis) and immunoreactive corticotropin-releasing factor (CRF) in the goldfish treated with dopamine antagonists

The in vivo effects of three dopamine (DA) antagonists on the cytology of the rostral pars distalis (RPD) were investigated in young goldfish ( Carassius auratus L.). Pimozide, sulpiride, and domperidone were injected for 5 (low dose, Experiment I) and 7 days (higher dose, Experiment II). Cytological and immunocytochemical techniques using antisera to (1–24) ACTH and (1–39) ACTH, human β-thyrotropin (TSHβ), and synthetic (1–41) CRF were applied to pituitary and brain sections. Cytometrical studies showed that the three drugs induced similar quantitative changes in the cells of the RPD. Prolactin (PRL)-secreting cell hypertrophy was significant in Experiment 11, whereas the nuclear enlargement was significant in both experiments. The numbers of cytoplasmic granules were similar in control and treated goldfish. Thyrotropic (TSH) cells and their nuclei were significantly enlarged in both experiments; their content in immunoreactive TSH was not clearly modified. Corticotropic (ACTH) cells showed significant nuclear and cellular hypertrophy, and labeled granules were often concentrated along the cell membrane. The amount of immunoreactive CRF present in the rostral neurohypophysial ramifications was reduced in the majority of treated fish. Solvent-injected controls showed no significant changes in the RPD. These results suggest that DA inhibits PRL cell activity in goldfish. TSH and ACTH cells appear stimulated by DA-receptor blockers, although differential effects on synthesis and release cannot be evaluated in in vivo experiments. A release of corticotropin-releasing factor may be involved in the ACTH cell stimulation. These data are compared with those obtained in other vertebrates.

Correlation of Na+,K+-ATPase content and plasma membrane surface area in adapted and de-adapted salt glands of ducklings.

During salt-water adaptation, an increase occurs in Na+,K+-ATPase content and surface area of the basolateral plasma membrane of the principal cell of the duck salt gland. To determine the degree to which these changes are correlated, accepted morphometric methods were used to determine numerical cell densities and plasma membrane surface densities of peripheral and principal cells. After adaptation, the plasma membrane surface area per principal cell was five times greater than in controls. Following de-adaptation, the plasma membrane content in principal cells returned to 1.9 times control levels. Two other cell constituents, mitochondria and lipid droplets, displayed similar quantitative changes. Na+,K+-ATPase content increased about fourfold with adaptation and decreased to near control levels with de-adaptation. Thus, changes in Na+,K+-ATPase content and basolateral plasma membrane surface area in adapting and de-adapting secretory epithelia of the salt gland occur nearly in parallel. These quantitative data enable Na+,K+-ATPase synthesis and degradation to be investigated in relation to membrane biogenesis.

Ethylene-Independent and Ethylene-Dependent Biochemical Changes in Ripening Tomatoes

Fruits of Lycopersicon esculentum Mill cv Sonatine stored in 6% CO(2), 6% O(2), and 88% N(2) for 14 weeks at 12 degrees C, exhibited a temporal separation of certain biochemical events associated with ripening.The specific activity of two citric acid cycle enzymes, citrate synthase and malate dehydrogenase, fell substantially during the first 2 weeks of storage when changes in organic acid concentration also occurred. During this period, lycopene, polygalacturonase, and ethylene were undetectable.When fruit were removed from store, ethylene was evolved and polygalacturonase and invertase activity were rapidly initiated as was synthesis of lycopene.To determine whether the changes in organic acid metabolism were affected by ethylene, fruit was kept at 22 degrees C in either a normal atmosphere or a normal atmosphere supplemented with 27 microliters per liter of ethylene, and it was shown that in both atmospheres similar quantitative changes to those described above occurred in the citric acid cycle enzymes specific activities before any detectable increase in the specific activities of invertase and polygalacturonase. These latter changes, together with pigment changes, occurred between 2 and 3 days earlier in fruit exposed to ethylene, compared with those kept in a normal atmosphere.

Rat endometrial bioelectric activity in vivo and in vitro: effects of adrenaline.

The transuterine potential difference (p.d.) generated by the endometrium was measured in vivo in anaesthetized, ovariectomized rats treated with 10 micrograms oestradiol-17 beta each day for 3 days. Intravenous injection of adrenaline induced transient hyperpolarization of the p.d.; sigmoid dose-dependency was observed over the range 2-40 micrograms/kg. Pre-treatment of the rats with syrosingopine shifted the dose-response curve to the left (enhancement) while combined propranolol (800 micrograms/kg) and phentolamine (800 micrograms/kg) shifted the dose-response curve to the right (depression). Isoprenaline gave similar quantitative changes in the p.d. in vivo which displayed a similar time course and sigmoid dose-dependency. Propranolol alone (800 micrograms/kg) shifted this dose-response curve significantly to the right. Addition of adrenaline or isoprenaline to the serosal solution bathing uteri incubated in vitro after removal from oestradiol-treated rats induced changes in endometrial bioelectric activity that exhibited sigmoid dose-response curves for the transuterine p.d. and short-circuit current (Isc). Propranolol significantly shifted the dose-response curves for adrenaline to the right. Only adrenaline caused small but significant decreases in the uterine resistance but these were not dose-dependent. Acetylcholine, 5-hydroxytryptamine, noradrenaline and phenylephrine had no effect on the transuterine p.d. in vivo or in vitro or on the Isc in vitro. No significant electrical changes could be induced by adrenaline or any of the other drugs across the uteri from untreated ovariectomized rats either in vivo or in vitro. The transluminal membrane p.d. of surface endometrial cells measured with micro-electrodes was depolarized by an addition of serosal adrenaline but only 41% of the cells successfully impaled responded in this way. The results indicate that endometrial cells under oestrogen stimulation possess electrogenic processes that can be modulated specifically by adrenaline through the mediation of a serosally sited beta-adrenoceptor. The physiological role of the mechanism has yet to be established.

8-Bromo-guanosine-3′,5′-monophosphate mimics the effect of acetylcholine on slow response action potential and contractile force in mammalian atrial myocardium

In isolated myocardium from the left atrium of guinea pigs, the effect of acetylcholine (ACh) and of 8-Br-cGMP on the slow response action potential (elicited after inactivation of the fast Na current) and on the isometric contractile force was studied. ACh reduced upstroke velocity and overshoot and abbreviated action potential duration. Though the sensitivity towards ACh (1.65 × 10 −8 m to 5.5 × 10 −6 m) widely differed, the decrease of upstroke velocity was accompanied in the most experiments by similar quantitative changes of contractile force. 8-Br-cGMP caused the same depression of the slow response action potential. The development of the negative inotropic effect was the more pronounced the stronger the upstroke velocity was depressed. 6 × 10 −4 m 8-Br-cGMP diminished upstroke velocity by 12 to 58% and isometric contractile force by 8 to 60%. The β-adrenergic compound isoproterenol abolished the inhibitory effects. But the absolute values of upstroke velocity, overshoot, and contractile force reached in the presence of isoproterenol were lower when the preparation had been pretreated with ACh or 8-Br-cGMP. It is concluded that cGMP might mediate the ACh effect on the surface membrane. The decline of upstroke velocity of the atrial slow response action potential suggests that 8-Br-cGMP could exert an inhibitory action on the Ca system of the membrane.

Changes in the soluble protein constitution of cucumber cotyledons following infection with two strains of cucumber mosaic virus

Changes in the soluble protein constitution of cucumber cotyledons following inoculation with two strains of cucumber mosaic virus ( W and N), which differe in the severity of symptoms they induce, were investigated by polyacrylamid gel electrophoresis and isoelectric focusing. Inoculation with either strain resulted in similar quantitative changes in soluble protein consitution as revealed by electrophoresis on 5, 7, 10 and 15% polyacrylamide gels. These changes were related both to the appearance of symptoms and of infective virus. Isoelectric focusing revealed an increase in a protein of pI approximately 4·0. Both strains elicited similar quantitative changes in peroxidase and polyphenoloxidase isoenzymes.