The modulation of calcium currents by the activation of mGluRs

Glutamatergic transmission in the central nervous system (CNS) is mediated by ionotropic, ligand-gated receptors (iGluRs), and metabotropic receptors (mGluRs). mGluRs are coupled to GTP-binding regulatory proteins (G-proteins) and modulate different second messenger pathways. Multiple effects have been described following their activation; among others, regulation of fast synaptic transmission, changes in synaptic plasticity, and modification of the threshold for seizure generation. Some of the major roles played by the activation of mGluRs might depend on the modulation of high-voltage-activated (HVA) calcium (Ca²⁺) currents. Some HVA Ca²⁺ channels (N-, P-, and Q-type channels) are signaling components at most presynaptic active zones. Their mGluR-mediated inhibition reduces synaptic transmission. The interference, by agonists at mGluRs, on L-type channels might affect the repetitive neuronal firing behavior and the integration of complex events at the somatic level. In addition, the mGluR-mediated effects on voltagegated Ca²⁺ signals have been suggested to strongly influence neurotoxicity. Rather different coupling mechanisms underlie the relation between mGluRs and Ca²⁺ currents: Together with a fast, membrane-delimited mechanism of action, much slower responses, involving intracellular second messengers, have also been postulated. In the recent past, the relative paucity of selective agonists and antagonists for the different subclasses of mGluRs had hampered the clear definition of the roles of mGluRs in brain function. However, the recent availability of new pharmacological tools is promising to provide a better understanding of the neuronal functions related to different mGluR subtypes. The analysis of the mGluR-mediated modulation of Ca²⁺ conductances will probably offer new insights into the characterization of synaptic transmission and the development of neuroprotective agents.     

Immunomicroscopical localization of human preformed natural antibodies against pig tissues in xenogeneic transplantation

Summary The targets of preformed natural antibodies need to be identified whenever the use of pig organs is considered for human transplantation. In this study we used extracorporeal perfusion of pig organs with human blood, immunocytological techniques and immuno-electron microscopy to identify the targets and the nature of human preformed natural antibodies against pig antigens. The antibodies were found to be mainly of the IgG and IgM type and directed not only against endothelial cells, but also against mesenchymal and epithelial structures. To reproduce an in vivo situation, a Bio-pump was used to xenoperfuse pig kidneys and livers with human fresh oxygenated blood at 37°C, drawn from polycythaemic patients. Biopsies showed a deposition of human IgG and IgM on tubuli and glomeruli of pig kidneys and on endothelial cells of pig livers. Preperfusion of pig liver with human blood for 45 minutes before perfusion of kidneys significantly reduced the deposition of the natural antibodies.     

Influence of irradiance on in vitro growth and proliferation of Vaccinium corymbosum (highbush blueberry) and subsequent rooting in vivo

The effects of light on in vitro proliferation and subsequent in vivo rooting and acclimatisation of Vaccinium corymbosum were investigated. The shoots were exposed in vitro to different irradiances (total radiation ranging from 55 to 240 μmol m⁻² s⁻¹) for 7 to 60 days. In vitro growth and proliferation and the possible consequences on in vivo rooting were observed.
As compared to the control treatment (55 μmol m⁻² s⁻¹), higher irradiances improved proliferation and rooting ratios only with short applications (7 days). Short but high (210 μmol m⁻² s⁻¹) exposures applied at the end of the proliferation phase increased in vivo growth and rooting of the shoots. The shoots treated with strong light for longer times (14 and 28 days) showed both inhibition of growth and red colour of leaves and sprouts, and were less vigorous when transferred in vivo.     

Neither Moderate Hypoxia nor Mild Hypoglycaemia Alone Causes Any Significant Increase in Cerebral [Ca2±i: Only a Combination of the Two Insults Has This Effect.: A 31P and 19F NMR Study

(1) The energy state and free intracellular calcium concentration ([Ca^2±_i) of super-fused cortical slices were measured in moderate hypoxia (～65 μM O₂), in mild hypoglycaemia (0.5 mM glucose), and in combinations of the two insults using ¹⁹F and ³¹P NMR spectroscopy. (2) Neither hypoxia nor hypoglycaemia alone caused any significant change in [Ca^2±_i. Hypoxia caused a 40% fall in phosphocreatine (PCr) content but not in ATP level, and hypoglycaemia produced a slight fall in both (as expected from previous studies). These changes in the energy state recovered on return to control conditions. (3) A combined sequential insult (hypoxia, followed by hypoxia plus hypoglycaemia) produced a 100% increase in [Ca^2±, and a decrease in PCr level to ～25% of control. The reverse combined sequential insult (hypoglycaemia, followed by hypoglycaemia plus hypoxia) had the same effect. On return to control conditions there was some decrease in [Ca^2±_i and a small increase in PCr content, but neither recovered to control levels. (4) Exposure of the tissue to the combined simultaneous insult (hypoxia plus hypoglycaemia) immediately after the control spectra had been recorded resulted in a fivefold increase in [Ca^2±_i and a similar decrease in PCr level to 20–25% of control. There was little if any change of [Ca^2±_i or PCr level on return to control conditions. (5) These results are discussed in terms of metabolic adaptation of some but not all of the cortical cells to the single type of insult, which renders the tissues less vulnerable to the combined insult.     

Pre- and Posttranslational Regulation of β-Endorphin Biosynthesis in the CNS: Effects of Chronic Naltrexone Treatment

Abstract: There appear to be two anatomically distinct β-endorphin (βE) pathways in the brain, the major one originating in the arcuate nucleus of the hypothalamus and a smaller one in the area of the nucleus tractus solitarius (NTS) of the caudal medulla. Previous studies have shown that these two proopiomelanocortin (POMC) systems may be differentially regulated by chronic morphine treatment, with arcuate cells down-regulated and NTS cells unaffected. In the present experiments, we examined the effects of chronic opiate antagonist treatment on βE biosynthesis across different CNS regions to assess whether the arcuate POMC system would be regulated in the opposite direction to that seen after opiate agonist treatment and to determine whether different βE-containing areas might be differentially regulated. Male adult rats were administered naltrexone (NTX) by various routes for 8 days (subcutaneous pellets, osmotic minipumps, or repeated intraperitoneal injections). Brain and spinal cord regions were assayed for total βE-ir, different molecular weight immunoreactive β-endorphin (βE-ir) peptides, and POMC mRNA. Chronic NTX treatment, regardless of the route of administration, reduced total βE-ir concentrations by 30–40% in diencephalic areas (the arcuate nucleus, the remaining hypothalamus, and the thalamus) and the midbrain, but had no effect on βE-ir in the NTS or any region of the spinal cord. At the same time, NTX pelleting increased POMC mRNA levels in the arcuate to ～ 140% of control values. These data suggest that arcuate POMC neurons are up-regulated after chronic NTX treatment (whereas NTS and spinal cord systems remain unaffected) and that they appear to be under tonic inhibition by endogenous opioids. Chromatographic analyses demonstrated that, after chronic NTX pelleting, the ratio of full length βE_1–31 to more processed βE-ir peptides (i.e., βE_1–27 and βE_1–26) tended to increase in a dose-dependent manner in diencephalic areas. Because βE_1–31 is the only POMC product that possesses opioid agonist properties, and βE_1–27 has been posited to function as an endogenous anatgonist of βE_1–31, the NTX-induced changes in the relative concentrations of βE_1–31 and βE_1–27/βE_1–26 may represent a novel regulatory mechanism of POMC cells to alter the opioid signal in the synapse.     

Injury and repair in biocide-treated spores of Bacillus subtilis

Abstract Bacillus subtilis NCTC 8236 spores exposed to appropriate concentrations of test biocides (glutaraldehyde, two iodine and two chlorine preparations) were able to repair injury if subsequently held in nutrient broth at 37°C but not in broth at 22°C, sterile filtered water at 4, 22 or 37°C or germination medium at 37°C. Repair appeared to occur primarily during outgrowth and was initiated soonest for iodine-treated spores and latest for glutaraldehyde-treated ones.     

FASTPAT: a fast and efficient algorithm for string searching in DNA sequences

A new string searching algorithm is presented aimed at searchingfor the occurrence of character patterns in longer charactertexts. The algorithm, specifically designed for nucleic acidsequence data, is essentially derived from the Boyer –Moore method (Comm. ACM, 20, 762 – 772, 1977). Both patternand text data are compressed so that the natural 4-letter alphabetof nucleic acid sequences is considerably enlarged. The stringsearch starts from the last character of the pattern and proceedsin large jumps through the text to be searched. The data compressionand searching algorithm allows one to avoid searching for patternsnot present in the text as well as to inspect, for each pattern,all text characters until the exact match with the text is found.These considerations are supported by empirical evidence andcomparisons with other methods.     

Habitat selection and space use overlap between feral horses,pronghorn, and greater sage-grouse in cold arid steppe

Populations of feral horses (Equus ferus caballus) in the western United States have increased during the past decade, consequently affecting co-occurring wildlife habitat. Feral horses may influence 2 native wildlife species, greater sage-grouse (Centrocercus urophasianus; sage-grouse) and pronghorn (Antilocapra americana) through mechanisms of habitat alteration and competition. Wyoming, USA, contains the largest populations of pronghorn and sage-grouse of any state and also has the highest degree of range overlap between feral horses and these species. Consequently, the effects that horses may have on pronghorn and sage-grouse populations in Wyoming have implications at local, state, and population-wide levels. Managers need information concerning habitat selection and space use overlap among these species to develop appropriate management strategies; yet this information is absent for most feral horse management areas. To address this knowledge need, we attached global positioning system (GPS) transmitters to horses, pronghorn, and sage-grouse within the greater Bureau of Land Management–Adobe Town Herd Management Area in southern Wyoming and northern Colorado, USA, between 2017 and 2021 to evaluate habitat selection and space use of all species during 3 biologically relevant seasons: spring (Apr–Jun; sage-grouse breeding, nesting, and early-brood rearing; pronghorn late gestation and early parturition), summer (Jul–Oct; sage-grouse summer and late-brood rearing; pronghorn late parturition and breeding), and winter (Nov–Mar; non-breeding season). Feral horses selected flatter slopes and shorter mean shrub height across all seasons and were closer to water in spring and summer. Pronghorn habitat selection was similar to horses, but they also avoided oil and gas well pads year-round. During spring, sage-grouse selected greater herbaceous cover, flatter slopes, and areas farther from well pads. In summer, sage-grouse selected greater mean shrub height, flatter slopes, and were closer to water. In winter, sage-grouse selected flatter slopes and areas with greater vegetation production during the preceding summer. Our results indicate strong year-round overlap in space use between horses and pronghorn, whereas overlap between horses and sage-grouse is greatest during the summer in this region. Consequently, managers should recognize the potential for horses to influence habitat quality of pronghorn and sage-grouse in the region.