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At least 307 records · Page 17Linked to original sources
PubMed2003

Motion characteristics of Murrah buffalo bull spermatozoa in various seasons and its relationship with functional integrity of the plasmallema.

Semen was collected from six adult (3.5-7-year-old) Murrah buffalo bulls at weekly intervals for 1 year and evaluated for routine parameters, motion characteristics, reactivity in hypoosmotic solution, and acrosomal and other morphological abnormalities of the spermatozoa. The overall motility (MOT), straight line velocity (VSL), curvilinear velocity (VCL), linearity (LIN), lateral head displacement (ALH) and average path velocity (VAP) were 66.85+/-2.79%, 26.58+/-0.24 and 107.07+/-1.47 microm/s, 26.91+/-0.01%, 11.19+/-0.09 and 61.78+/-2.79 microm/s, respectively. Significant seasonal variation was observed in sperm kinematics and hypoosmotic swelling (HOS) reactivity. Except for LIN, the mean values of sperm dynamics were higher during summer and rainy season and significantly lower in winter season. Sperm kinematics showed significant (P<0.01) positive correlation (r=0.25-0.60) with plasmallemal integrity. Ejaculates with less than 50% HOS-reactive spermatozoa had significantly lowered MOT, VSL, VCL and VAP as compared to the ejaculates with >50% HOS-positive spermatozoa. No significant difference was observed in sperm kinematics among the ejaculates having 50-70% and >70% HOS-reactive spermatozoa. The trend of motion dynamics of the spermatozoa with respect to HOS reactivity was similar in all the three seasons (summer, rainy and winter). The results indicate that ejaculates having more than 50% of HOS-reactive sperm show a higher magnitude of sperm kinematics compared to ejaculates having less than 50% HOS-positive spermatozoa.

Animals↗
PubMed2002

Vaccinia virus complement control protein is monomeric, and retains structural and functional integrity after exposure to adverse conditions.

Vaccinia virus complement control protein (VCP) possesses the ability to inhibit both classical and alternative pathways of complement activation, as well as bind to heparin or heparan sulfate proteoglycans, making it a unique multifunctional protein with therapeutic potential. Recently, the structure of the complete molecule of VCP was determined by X-ray crystallography. Two or three VCP molecules were packed within the unit cells of both crystal forms. Using gel filtration, VCP has now been shown to exist as a monomer in solution. To test the stability of this molecule, VCP was studied by nuclear magnetic resonance (NMR) over a range of temperatures and by differential scanning calorimetry (DSC). It was also subjected to adverse physical conditions, including, freeze-thawing, changes in pH, changes in temperature, and storage at room temperature. VCP melts fully reversibly, and it maintained its 3-D structure and the ability to inhibit serum-induced hemolysis of sheep red blood cells after exposure to many extreme conditions. The robustness of VCP may be rationalized in terms of its architecture.

Animals↗
PubMedDate not supplied

Functional integration of the transport of ammonium, glutamate and glutamine in astrocytes.

This short review surveys the effects of extracellular potassium, released by neuronal activity, on the fluxes of ammonium, glutamate and glutamine in astrocytes. There is evidence that each of these fluxes is modulated by potassium-induced changes in astrocytic pH. The result is viewed as an integrated response to neuronal activity. The unusually high permeability of astrocyte cell membrane to ammonium ions, together with the normal transmembrane gradient of pH, enables astrocytes to accumulate ammonium appreciably. However, at loci of neuronal activity, effective ammonium ion permeability is diminished and the cytosol is alkalinized, resulting in a local decline in intracellular ammonium concentration. Intracellular potassium concentration rises at these same loci, creating the conditions for a 'potassium-ammonium countercurrent' in which ammonium ions migrate intracellularly towards sites of neuronal activity as potassium ions diffuse away. Physiologic elevations of extracellular potassium evoke a marked 'paradoxical' increase in the velocity of glutamate uptake in astrocytes. This increase correlates well with the extent of potassium-induced alkalinization. Further, recent evidence identifies a major transporter of glutamine in astrocytes (System N) as a glutamine/proton exchanger. Potassium can reverse the transmembrane gradient of protons in astrocytes, and increase intracellular glutamine concentration, creating the conditions for a reversal of glutamine flux via System N from uptake to export. These flux changes, evoked by potassium released from active neurons, combine to accelerate glutamate-glutamine cycling.

Animals↗
PubMed1986

Integrated functioning of the chloroplast coupling factor.

This review is focused on some functional characteristics of the chloroplast coupling factor. The structure of the enzyme and the putative role of its subunits are recalled. An attempt is made to discriminate the driving force and the activator effects of the electrochemical proton gradient. Respective roles of delta pH, delta phi, external and internal pH are discussed with regard to mechanistic implications. The hypothesis of a functional switch of the enzyme between two states with better efficiency either in ATP synthesis or in ATP hydrolysis is also examined. A brief survey is made on some problems complicating quantitative studies of energy coupling, such as localized chemiosmosis, delta pH and delta phi computations, and scalar ATPases. The main data on the enzyme activation and the energy-dependent release of tightly bound nucleotides are summarized. The arguments for and against the catalytic competence of theses nucleotides are reviewed. Lastly, some prevailing models of the catalytic mechanism are presented. The relevance of nucleotides binding change events in this process is discussed.

Adenosine Diphosphate↗
PubMed2002

Towards integrating functional imaging in the treatment of prostate cancer with radiation: the registration of the MR spectroscopy imaging to ultrasound/CT images and its implementation in treatment planning.

PURPOSE: Dose-escalation to intraprostatic tumor deposits detected by magnetic resonance spectroscopy (MRS) is an example of tumor-targeted radiation therapy. Because treatment planning for prostate brachytherapy is performed based on ultrasound (US)/computed tomography (CT) images, a sine qua non of this technique is the ability to map MRS-positive volumes (obtained in a gland deformed by the endorectal balloon coil) to the US/CT images. An empirical algorithm designed to perform this function, and its validation, are described. METHODS AND MATERIALS: Mathematically, the problem of mapping points between the MR and US/CT domains comes to: (a) ascertaining that the position of any point in the interior of the prostate is uniquely determined by the shape of the gland, and (b) finding an algorithm that describes this relationship. The image registration algorithm described here is based on the assumption that points within the gland maintain the same relative position with respect to both the axial contours of the prostate and the center of the prostate along the superior-inferior direction. Relative positions of MRS-positive voxels are calculated with this method in both MR and US/CT space. For a particular voxel in the MR space, one obtains first the z coordinate in the US/CT space, that is, along the superior-inferior direction. This determines the axial slice in the US/CT frame of reference where the other two coordinates (x, y) will be calculated. The validity of this algorithm was examined with the aid of a pelvic phantom built to simulate realistically the prostate and its surrounding bony and tissue structures and with CT scans of implanted patients obtained, at several weeks' intervals, as part of an edema-resolution study. Seventy-five "dummy" seeds were placed in the phantom, within the simulated prostate gland, in a quasi-regular pattern. The coordinates of these seeds were determined and thus served as markers of prostate deformation when an inflated rectal probe was introduced in the phantom. CT images of this phantom were taken for different volumes of the MR rectal probe and in each case the prostate outlines were contoured and seed coordinates calculated. Using these data, the predictions of the mapping algorithm could be directly verified. RESULTS: Absolute values of the 3D-positional errors in this algorithm were 2.2 mm +/- 1.2 mm (average +/- SD). Only 6 of 75 seeds had positional displacement of 4 mm or more. Similar results were obtained in the patient analysis. CONCLUSIONS: In comparison to the MRS voxel size (6.25 x 6.25 x 3.0 mm3), the present algorithm achieves the desired clinical accuracy. As well, with this 3D algorithm seed positions are reconstructed with an uncertainty that, along the z direction, is less than half the thickness of the typical US slice (0.5 cm).

Algorithms↗
PubMed1996

Studies on integrative functions of the human frontal association cortex with MEG.

Our MEG studies on the human frontal association cortex are briefly reviewed. (1) The no-go potential was first found at go/no-go reaction-time hand movement task with discrimination between different colour light stimuli in the prefrontal cortex of monkeys. The potential was recorded in human subjects with EEG over the scalp, but its current dipoles could be localized only by use of MEG, in the dorsolateral part of the frontal association cortex in both cerebral hemispheres. The function for no-go decision and subsequent suppressor action was thus substantiated in the human frontal cortex. (2) Utterance of a short noun in Japanese was found to be initially preceded by an activity in the lower lateral part of the frontal lobe and then by that around the central sulcus. The area of the former, often in both hemispheres, appears to correspond to Broca's motor speech centre and that of the latter, always in both hemispheres, to correspond to the motor-somatosensory cortices. (3) Intensive and continuous concentration on mental calculation and some 'abstract' thinking for a few minutes were often associated with magnetic theta (5-7 Hz) wave bursts in the frontal part of the scalp. Dipole fitting suggested that the electrical current dipoles occur successively and scattered in wide areas of the frontal lobe on both sides. They are to be called "frontal mental theta wave", revealing dynamic and active participation of the frontal lobe in mental functions.

Adult↗
PubMed2000

Perspectives on the integrative functions of the 'sympatho-adrenomedullary system'.

An historic survey is given of the gradual change of views and concepts concerning how the sympatho-adrenomedullary system is organized and operates: While it for nearly a century was considered to merely exhibit more or less generalized activation-inhibition responses, experimental studies during the last 50 years have revealed how it instead constitutes a highly sophisticated instrument for control, engaged in a variety of differentiated response patterns by which the brain controls events in major organ systems, down to include their cellular-molecular levels of organisation.

Adrenal Medulla↗
PubMed1986

Integrated function of a kinetic proofreading mechanism: double-stage proofreading by isoleucyl-tRNA synthetase.

Experimental measurements for isoleucyl-tRNA synthetase proofreading valyl-tRNAIle in Escherichia coli previously have been incorporated into the conventional Michaelis-Menten model for this system. This model was augmented to include two stages of proofreading--the aminoacyl adenylate and aminoacyl-tRNA stages--and used to predict the values of four additional rate constants that have been determined experimentally. The results suggest that two stages of conventional kinetic proofreading with binding sites designed for isoleucine (the "correct" substrate) are inconsistent with the experimental data, that a double-stage mechanism in which one stage (the "double-sieve") involves a binding site designed for valine (the "incorrect" substrate) and the other involves a binding site designed for isoleucine is consistent with all the experimental data, and that the experimental data are not sufficiently accurate to distinguish the stage at which the double-sieve mechanism operates in vivo. Furthermore, analysis of the model suggests that four parameters have the most questionable values and that experimental refinement of their estimates will be needed to determine which of the two stages involves the double-sieve mechanism.

Amino Acyl-tRNA Synthetases↗
PubMedDate not supplied

Direct evidence for an integrated function of J chain and secretory component in epithelial transport of immunoglobulins.

J chain is a polypeptide of molecular weight (Mr) approximately 15,000 common to human dimeric IgA and pentameric IgM. These immunoglobulin polymers show a high affinity for secretory component (SC) in vitro, a feature that, in some studies, has been claimed to be a function of the J chain. SC is a glycoprotein of Mr approximately 80,000 which is expressed on the basolateral surfaces of secretory epithelial cells where, according to a current hypothesis, it may act as a receptor for dimeric IgA and pentameric IgM which are selectively transported through secretory epithelial cells into exocrine fluids. Previous studies, however, have not excluded the possibility that secretory cells express isotype-specific Fc receptors for IgA and IgM which may be involved in epithelial transport. We now report that the adsorption of immunoglobulin polymers to SC-expressing epithelial cells depends solely on a J chain-determined binding site. This finding lends biological significance to the striking J-chain expression shown by immunoglobulin-producing immunocytes in secretory tissues.

Biological Transport↗
PubMed1999

Release properties and functional integration of noradrenergic-rich tissue grafted to the denervated spinal cord of the adult rat.

Noradrenaline- (NA-) containing grafts of central (embryonic locus coeruleus, LC) or peripheral (juvenile adrenal medullary, AM, autologous superior cervical ganglionic, SCG) tissue were implanted unilaterally into rat lumbar spinal cord previously depleted of its NA content by 6-hydroxydopamine (6-OHDA) intraventricularly. A microdialysis probe was implanted in the spinal cord 3-4 months after transplantation, and extracellular levels of noradrenaline were monitored in freely moving animals during basal conditions and following administration of pharmacological or behavioural stimuli. Age-matched normal and lesioned animals both served as controls. Morphometric analyses were carried out on horizontal spinal sections processed for dopamine-beta-hydroxylase (DBH) immunocitochemistry, in order to assess lesion- or graft-induced changes in the density of spinal noradrenergic innervation, relative to the normal patterns. In lesioned animals, the entire spinal cord was virtually devoid of DBH-positive fibers, resulting in a dramatic 88% reduction in baseline NA, compared with that in controls, which did not change in response to the various stimuli. LC and SCG grafts reinstated approximately 80% and 50% of normal innervation density, respectively, but they differed strikingly in their release ability. Thus, LC grafts restored baseline NA levels up to 60% of those in controls, and responded with significantly increased NA release to KCl-induced depolarization, neuronal uptake blockade and handling. In contrast, very low NA levels and only poor and inconsistent responses to the various stimuli were observed in the SCG-grafted animals. In AM-grafted animals, spinal extracellular NA levels were restored up to 45% of those in controls, probably as a result of nonsynaptic, endocrine-like release, as grafted AM cells retained the chromaffine phenotype, showed no detectable fibre outgrowth and did not respond to any of the pharmacological or behavioural challenges. Thus, both a regulated, impulse-dependent, and a diffuse, paracrine-like, NA outflow may play roles in the recovery of lesion-induced sensory and/or motor impairments previously reported with these types of grafts following transplantation into the severed spinal cord.

Age Factors↗
PubMed2003

Selective depletion of fibroblasts preserves morphology and the functional integrity of peritoneum in transgenic mice with peritoneal fibrosing syndrome.

BACKGROUND: A peritoneal fibrosing syndrome (PFS) can progressively reduce peritoneal ultrafiltration during chronic peritoneal dialysis in patients with renal failure. The pathogenesis of PFS is unclear and the role of peritoneal fibroblasts has not been evaluated experimentally. METHODS: We followed the fate of fibroblasts producing PFS in a mouse model using fibroblast-specific protein 1 (FSP1) as a marker. PFS was induced by daily peritoneal infusions of chlorhexidine gluconate (CHG) saline into transgenic mice expressing the thymidine kinase (Delta tk) gene under the control of the FSP1 promoter (FSP1.Delta tk mice). To demonstrate the role of fibroblasts in PFS, we treated these FSP1.Delta tk mice with a nucleoside analogue to induce DNA chain termination and fibroblast death. RESULTS: Mice receiving peritoneal infusions of CHG saline every other day for 2 weeks developed increasing numbers of FSP1+ fibroblasts in the subserosal layers of the visceral peritoneum. Mac-3+ monocytes (macrophages) subsequently accumulated over the next 2 weeks in association with increased deposition of type I collagen and increased endothelial vascularity (CD31+) in these subserosal tissues. Since these peritoneal fibroblasts expressed monocyte chemoattractant protein-1 (MCP-1), heat shock protein 47 (HSP47), and vascular endothelial growth factor (VEGF), we suspect they were partially responsible for macrophage recruitment, matrix production, and the neoangiogenesis in the subserosal tissue. Treatment of PFS in FSP1.Delta tk transgenic mice with a nucleoside analogue selectively reduced the numbers of peritoneal fibroblasts and attenuated the attendant changes in peritoneal histology. Rescuing the peritoneal membrane from chronic thickening and neoangiogenesis by reducing the number of fibroblasts also preserved ultrafiltration. CONCLUSION: Peritoneal fibroblasts play a pivotal role in PFS, and their deletion using a fibroblasts-specific transgene was effective in preventing peritoneal fibrogenesis.

Animals↗
PubMed2004

Biliverdin protects the functional integrity of a transplanted syngeneic small bowel.

BACKGROUND & AIMS: Heme oxygenase-1 (HO-1) protects against inflammation in many disease models. By degrading heme, HO-1 generates carbon monoxide (CO), iron and biliverdin. We investigated whether biliverdin would protect rat syngeneic small intestinal transplants (SITx) against damage and, if so, by what mechanism. METHODS: Motility was assessed by organ bath techniques. Inflammatory cytokines and mediators were assessed by RT-PCR and spectrophotometric assays. Myeloperoxidase histochemistry for neutrophils was performed in jejunal segments. Western blots were performed for biliverdin reductase and HO-1 expression. Permeability was expressed as the mucosal to serosal clearance of fluorescent dextran in everted gut sacs. NF-kappaB activation was assessed via EMSA. RESULTS: Biliverdin significantly improved survival of recipients following SITx after prolonged intestinal ischemia (6 hours). Biliverdin treatment (1) led to a significant decrease in mRNA expression of iNOS, Cox-2, and ICAM-1 as well as the inflammatory cytokines IL-6 and IL-1beta; (2) decreased neutrophil infiltration into the jejunal muscularis; and (3) prevented SITx-induced suppression of intestinal circular muscle contractility. CONCLUSIONS: Biliverdin administration attenuates transplantation-induced injuries to the small bowel by its anti-inflammatory action. Importantly, biliverdin enhanced recipient survival. A comparison of the mechanisms by which biliverdin exerted these salutary effects compared with inhalation of CO, which we previously showed had salutary effects, suggests that the 2 compounds (biliverdin and CO) exert their effects in part by different mechanisms. This implies that the different products of HO-1 action on heme may exert protective effects that are additive or synergistic.

Animals↗
PubMed1999

Fate and functional integrity of fresh and frozen-thawed ram spermatozoa following intrauterine insemination.

Ewes in a synchronized oestrus were inseminated (intrauterine) with fresh and frozen-thawed spermatozoa and the spermatozoa were either recovered from each section of the reproductive tract after the animal was killed (Experiments 1a and 1b) or after they were voided from the cervix (Experiment 2). In Experiment 1a, only 1.2+/-0.27% of the original inseminate was recovered. Placing a ligature at the base of the uterine horn in Experiment 1b led to the recovery of 3.0+/-0.33% of the original inseminate, located mainly in each uterine horn (33.1+/-5.48%), and each isthmic and ampullary region of the oviduct (2.9+/-5.48% and 4.0+/-5.48%, respectively). A higher proportion of spermatozoa recovered from the isthmus were uncapacitated when observed by chlortetracycline staining than those recovered from the uterus (26.4+/-1.92% and 15.6+/-1.92%, respectively, P<0.05). Experiment 2 showed that large proportions of spermatozoa were voided from the tract through the vagina, with similar numbers of fresh and frozen-thawed spermatozoa lost from the tract. However, frozen thawed spermatozoa were lost at a faster rate than fresh (P<0.05) and with a more advanced membrane state (66.8+/-1.30% and 53.2+/-1.30% were acrosome reacted respectively; P<0.001). Large numbers of recovered spermatozoa had lost their tails, with frozen-thawed spermatozoa more susceptible to tail loss than fresh spermatozoa (55.0+/-0.96% and 45.5+/-0.96% respectively; P<0.05).

Animals↗
PubMed2005

An integrated functional genomics and metabolomics approach for defining poor prognosis in human neuroendocrine cancers.

Human neuroendocrine (NE) cancers range from relatively indolent to highly aggressive. In this study, we combine functional genomics with metabolomics to identify features of NE cancers associated with a poor outcome. Analysis of GeneChip datasets of primary prostate tumors, as well as lymph node and liver metastases from transgenic mice with a NE cell cancer, plus derived NE cell lines yielded a signature of 446 genes whose expression is enriched in neoplastic mouse prostatic NE cells. This signature was used for in silico metabolic reconstructions of NE cell metabolism, directed liquid chromatography/tandem MS analysis of metabolites in prostatic NE tumors and cell lines, and analysis of GeneChip datasets of human NE tumors with good or poor prognoses. The results indicate that a distinguishing feature of poor-prognosis NE tumors is a glutamic acid decarboxylase-independent pathway for production of GABA and a pathway for production of imidazole-4-acetate that involves dopa decarboxylase and a membrane-associated amine oxidase, amiloride-binding protein 1. Electrophysiological studies disclosed that imidazole-4-acetate can bind and activate GABA(A) receptors expressed by transformed NE cells, thus providing a previously uncharacterized paradigm for NE tumor cell signaling. Transcriptional, metabolic, and electrophysiologic features of transformed mouse NE cells are also evident in neural progenitor cells.

Animals↗