Response of otolith-related neurons in bilateral vestibular nucleus of acute hemilabyrinthectomized cats to off-vertical axis rotations.
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Biomedical subjects
Publications and source records attributed to Y S Chan.
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Cymbidium mosaic virus (CyMV) is the most prevalent virus infecting orchids. Here, we report the isolation of partial cDNA clones encoding the genomic RNA of CyMV. Like most of the polyadenylated monopartite positive-strand RNA viruses, the open reading frame (ORF) coding for the viral coat protein (CP) is located at the 3' end. The ORF predicts a polypeptide chain of 220 amino acids with a molecular weight of 23,600. Sequence comparison of this ORF to the CP sequences of potato virus X(PVX) and white clover mosaic virus (WClMV) revealed a strong amino acid homology in the mid-portion of the CP, but the overall homology was low. The CyMV CP gene was placed downstream of a cauliflower mosaic virus 35S promoter and the chimaeric gene was transferred into Nicotiana benthamiana. Transgenic plants expressing the CyMV CP were protected against CyMV infection.
In female pentobarbital-anesthetized Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR), changes in spontaneous discharges of cardiovascular neurons in the rostral ventrolateral medulla (RVL) in response to iontophoretic application of angiotensin II (Ang II) were studied and compared. It was found that iontophoretic application of Ang II to RVL increased the spontaneous neuronal activities of 30% of the cardiovascular neurons in both types of rats and that the increase was significantly greater in SHR than in WKY. In both types of rats, there was an increase in arterial blood pressure in response to iontophoretic release of Ang II to RVL. The pressor response was accompanied by tachycardia, which was significantly greater in SHR than in WKY. The present study provides evidence that Ang II acts directly on cardiovascular neurons in RVL, and in SHR, an enhanced sensitivity and responsiveness of the RVL cardiovascular neurons to Ang II may augment the sympathetic outflow from RVL and contribute to the genesis of hypertension.
Single unit activities were recorded from the rostral ventrolateral medulla (RVL) of pentobarbital-anesthetized normotensive Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Throughout the recording period, arterial blood pressures of WKY (mean arterial pressure, MAP = 103.1 mm Hg) and SHR (MAP = 159.2 mm Hg) remained stable at the respective basal levels. The units recorded in this study were all spontaneously active and cardiac-locked. Two types of discharge patterns, namely single and double discharges, were identified. These single and double discharge units were found to distribute randomly in RVL. In WKY, 92.6% of RVL neurons exhibited single discharges whereas in SHR, the majority (57%) of RVL neurons exhibited double discharges. The mean firing rate of single discharge units in RVL of SHR was significantly higher than that of WKY, whereas the mean firing rate of double discharge units in WKY was similar to that of SHR. About half of the units studied were also tested for antidromic collision; all units tested could be antidromically activated from the intermediolateral column (IML) of the thoracic spinal cord and the lowest threshold sites were consistently localized within IML. In both groups of rats, the axonal conduction velocity of RVL neurons showed a bimodal distribution viz. the fast and slow conducting axons. The mean conduction velocities of each of these two groups of neurons in WKY and SHR were similar. Most of the double discharge units in WKY and SHR belonged to the fast conducting type.(ABSTRACT TRUNCATED AT 250 WORDS)
Nucleotide sequencing data from a region extending 35 kb inward from the right inverted terminal repeat (ITR) of the vaccinia virus (VV) genome established the presence of VV homologues of the Shope fibroma virus (SFV) ITR proteins. The nucleotide sequences, comprising a total of 8.6 kb, and the amino acid translations for nine predicted open reading frames (ORFs) (designated SalF4L, SalF 19R, SalF21R, B4R, B8R, B9R, B10R, and B14R) are presented. Eight of the nine VV genes and all the SFV ORFs are transcribed towards their genomic termini. However, the relative positions of the VV genes (genus Orthopoxvirus) are different than those of the corresponding ORFs in SFV (genus Leporipoxvirus), indicating complex rearrangements of DNA in the genome of one or both of these viruses subsequent to their divergence from a common ancestor. Several other features of the VV ORFs were noted. SalF4L, B7R, B8R, and B9R have hydrophobic amino-terminal signal sequences but lack discernible membrane anchor domains suggesting that the proteins may be secreted. VV ORF SalF19R has a single cysteine-rich region homologous to the multiple domains of nerve growth factor receptor (NGFR), CD40, OX40 (a glycoprotein from the surface of activated murine T lymphocytes), and the recently described tumor necrosis factor receptors. Just downstream of the ORF SalF19R and in a different reading frame, there are another two related cysteine-rich domains, indicating that SalF19R was once a larger gene. B4R has homology to the host range gene of cowpox virus and to related genes near the opposite end of the vaccinia virus genome, and contains regions homologous to the repeat domains of erythrocyte ankyrin. In addition, several of the VV ORFs have homology to ORFs from near the opposite end of the VV genome, thus increasing the number of known VV gene families.
The structures of two vaccinia virus genes (B15R and B18R) from near the right inverted terminal repeat are described. These genes encode proteins of 36.5K and 40.7K, respectively, which have an N-terminal hydrophobic sequence, possible sites for attachment of N-linked carbohydrate and a short string of hydrophobic residues near the C terminus. These properties are consistent with the mature proteins being either virion, cell surface or secretory glycoproteins. Protein sequence comparisons established that the two gene products are related to each other (20% identity) and to the immunoglobulin (Ig) superfamily. Intriguingly, the nearest homologues of these proteins in the SWISS-PROT (version 14) database are the human and murine interleukin-1 receptors, although both proteins are related to a wide range of Ig superfamily members, including the interleukin-6 receptor. The product of one of these genes is known to be expressed on the cell surface early during infection and immunity directed against it confer resistance to virus infection without directly neutralizing virus infectivity. We propose a novel method for virus immune evasion in which the product of one or both of these proteins may bind interleukin-1 and/or interleukin-6 and prevent these cytokines reaching their natural receptors. In consequence the inflammatory response would be diminished and virus replication enhanced.
The nucleotide sequence of 42090 bp of vaccinia virus strain WR is presented. The sequence includes the SalI L, F, G and I fragments and starts near the centre of the HindIII A fragment and extends rightwards towards the genomic terminus, finishing approximately 0.5 kb internal of the inverted terminal repeat (ITR). Translation of this region has identified 65 open reading frames (ORFs) of greater than 65 amino acids in length. Fifty-one of these which do not extensively overlap other larger ORFs have been subjected to further analysis; the other 14 are termed minor ORFs. In the rightmost 28.7 kb, the genes are, with one exception, transcribed towards the genomic terminus, similar to the arrangement of genes at the left end of the virus genome. Internal of this region the genes are expressed off either DNA strand but still predominately rightwards. ORFs are tightly packed with few intergenic non-coding regions of greater than 250 bp. Protein sequence comparisons have established a remarkably high number of homologies with entries in existing protein databases. Of these, DNA ligase, thymidylate kinase, two serine-threonine protein kinases, two serine proteinase inhibitors (serpins), two interleukin-1 receptor homologous and a discontinuous ORF related to tumour necrosis factor receptor have been reported. Other homologies include lectins, profilin, 3 beta-hydroxy steroid dehydrogenase, superoxide dismutase, guanylate kinase, ankyrin and complement factor H. In addition, there are a number of polypeptides with predicted properties of membrane-associated, secretory or glyco-proteins. Twelve gene families are described here and elsewhere. There is considerable similarity between genes from the right and left end of the virus genome that may have arisen by terminal transposition events. Several differences from the corresponding region of vaccinia virus strain Copenhagen sequence are noted. Near the right terminus the sequences diverge completely, and internal of this there are multiple examples of deletion of short sequences (eight to 10 nucleotides) that lie within penta- or hexanucleotide direct repeats.
The plasma melatonin in the confluens sinuum of anesthetized rabbits and rats exhibited an episodic release pattern, with pulses superimposed on a basal level. The rhythmicity of pineal melatonin release revealed by our experimental paradigm was dependent on the blood sampling interval. In rabbits, it was found that the plasma melatonin level in the confluens sinuum was 7-15 times higher than that obtained from the plasma collected at the same time from the peripheral artery where the melatonin level also showed a pulsatile pattern. Diurnal variation in pineal melatonin level in the confluens sinuum was observed only in acutely blinded rabbits and rats. Electrical stimulation of the unilateral cervical sympathetic trunk of rabbits resulted in elevation of the level of plasma melatonin in the confluens sinuum, while the frequency of pulse peaks of plasma melatonin revealed at 4-min intervals was not affected. Similar results were obtained in rabbits with systemic administration of beta-adrenergic agents. These findings suggest that the release of pineal melatonin into the confluens sinuum is under the regulation of the cervical sympathetic system. The sympathetic influence on pineal melatonin secretion seems to vary with age. Furthermore, age-related fluctuation of systemic blood melatonin appeared to be the results of changes in the rate of pineal melatonin secretion, body growth and ageing.
To investigate whether neurons in the medullary reticular formation of decerebrate cats can encode spatial orientation during 360 degrees off-vertical axis rotations (OVAR), responses of these units were studied during constant velocity OVAR in clockwise (CW) and counter-clockwise (CCW) directions. Tilt-sensitive units, some of which were activated by antidromic stimulation of the thoracic cord, were found to respond to 360 degrees OVAR with position-and direction-dependent modulation in discharge rates. For the individual responsive unit there was a difference in the spatial location of the CW and CCW discharge maxima, while a comparable response gain was obtained for rotations in either direction. The orientation of the best response of each unit was not significantly modified by changes in the amplitude of head tilt and/or velocity of rotation. Such characterized directional axes of the medullary reticular units were found to have a fairly equal distribution over the 360 degrees. The role that neurons in the medullary reticular formation plays in encoding otolith inputs with respect to head movements is discussed.
Cardiovascular responses to electrical microstimulation of the ventrolateral medulla were investigated in both Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) under pentobarbital anesthesia. The threshold intensity required to elicit a change in blood pressure (BP) and the cardiovascular responses in these two groups of rats upon electrical stimulation were compared. It was found that the region with the lowest threshold intensities was located in the rostral ventrolateral medulla (RVL) and the threshold intensities were much lower in SHR than in WKY. Electrical stimulation of this brain region also resulted in a greater increase in BP during stimulation in SHR, compared to control. In SHR, upon termination of stimulation, the BP dropped to a level above the pre-stimulation level and this was followed by a prolonged, sustained elevation in BP before returning to the control level, whereas in WKY, the BP showed an initial drop to below the pre-stimulation level and then returned to the control level. These results suggest an enhanced responsiveness to electrical stimulation in SHR. Although the heart rate (HR) increased to a similar extent during electrical stimulation in both groups of rats, upon termination of stimulation WKY exhibited bradycardia followed by tachycardia before the HR returned to the pre-stimulation level, whereas SHR exhibited tachycardia which was maintained for a substantial period of time before returning to the pre-stimulation level. The results suggest a lower baroreceptor sensitivity in SHR. The change in HR/change in BP was smaller in SHR than in WKY, suggesting that the increase in HR may not contribute to the increase in BP during stimulation as much in SHR as in WKY.(ABSTRACT TRUNCATED AT 250 WORDS)
The level and release pattern of plasma melatonin in the confluens sinuum of 28 sighted and 18 blinded (i.e. acute bilateral orbital enucleation) rabbits anesthetized with pentobarbital were studied. The animals had been adapted to a 12:12 h light-dark regime. Blood samples were collected from the cannulated confluens sinuum and/or the femoral artery at either 2- or 4-min intervals in both the light and dark phases. Plasma melatonin was determined by radioimmunoassay. In all rabbits studied, plasma melatonin in the confluens sinuum exhibited an episodic release pattern, with pulses superimposed on a basal level. At 4-min sampling intervals, the pulsatile release of melatonin in sighted rabbits were 3.8 peaks/h in the light phase; shorter sampling interval (2-min) revealed more frequent pulsatile release of melatonin (5.1 peaks/h). In sighted animals in the light phase, the level of melatonin in the plasma of confluens sinuum was 7-15 times higher than that obtained from the plasma collected at the same time from the peripheral artery where the level of melatonin also exhibited pulsatile pattern. In blinded animals, melatonin levels in terms of mean concentration, mean maximum level and mean minimum (or baseline) level obtained in the dark phase were 12-14 times higher than those obtained in the light phase. These results suggest that the level of melatonin exhibits a diurnal rhythm in the confluens sinuum of rabbits.
Nucleotide sequencing of the vaccinia virus SalI F DNA fragment identified an open reading frame of 552 amino acids encoding a protein of 63.3 kDa. The deduced amino acid sequence shares 30% identity with S. pombe and S. cerevisiae DNA ligases, with homology strongest near the carboxy terminus and around the lysine residue required for ligase-adenylate formation. Prokaryotic DNA ligases are poorly related to the vaccinia sequence. The initiation codon of the ORF forms part of a late transcriptional initiation sequence TAAATG and is preceded by two overlapping early transcriptional termination signals, TTTTTTTAT. Nonetheless, RNA mapping showed that the ligase gene is transcribed early during infection and the 5' end of the mRNA maps to the TAAATG motif. The possible roles of a DNA ligase in vaccinia virus DNA replication and recombination are discussed.
The influence of electrical stimulation of the unilateral sympathetic trunk on the release patterns of plasma melatonin in the confluens sinuum of anesthetized rabbits was studied. Plasma melatonin exhibited an episodic release pattern, with pulses superimposed on a basal level. The mean concentration, mean maximum and minimum levels of plasma melatonin in the confluens sinuum were found to be significantly elevated when the sympathetic trunk was electrically stimulated. The frequency of pulse peaks of plasma melatonin sampled at 4-min intervals was not affected by electrical stimulation. Our results suggest that the diurnal rhythmic secretory pattern of pineal melatonin into the confluens sinuum of rabbits is regulated by the cervical sympathetic system.
The nucleotide sequence and deduced amino acid sequence of a vaccinia virus gene from the SalI F fragment are shown. The predicted polypeptide shares 42% amino acid identity over a 200 amino acid region with Saccharomyces cerevisiae thymidylate kinase (TmpK) and has low homology with herpes simplex virus deoxypyrimidine kinase. Northern blotting and S1 nuclease protection showed that the TmpK gene is transcribed early during infection and mapped the mRNA 5' end to immediately upstream of the second inframe ATG codon of the open reading frame (ORF). The encoded polypeptide is predicted to be 204 amino acids long (23.2 kD) and is almost colinear with yeast TmpK. Vaccinia virus possesses genes for TK and TmpK, separated by 57 kilobases of DNA, which are co-ordinately expressed and the encoded enzymes perform sequential steps in the same biochemical pathway.
Nucleotide sequencing of a region of the vaccinia virus genome proximal to the right inverted terminal repeat (ITR) identified two open reading frames (ORFs) encoding proteins of 39K and 40K with amino acid homology to each other, to another vaccinia virus gene near the opposite end of the virus genome and to the superfamily of serine proteinase inhibitors (serpins). Serpins have now been found in poxviruses from the genera orthopox (cowpox and vaccinia viruses), leporipox (myxoma virus) and avipox (fowlpox virus). One of the vaccinia virus serpins identified here (B13R) shares 92% amino acid identity with the serpin from cowpox virus and 46% and 19% identity with vaccinia serpins B24R and K2L, respectively. The amino acid sequence of B13R reported here differs at 11 positions from a recently reported sequence and contains an additional three internal residues. The serpin genes near the right ITR are separated by 8 kb of DNA. Both genes contain early transcriptional termination signals just downstream of the ORFs and are transcribed in a rightward direction towards the end of the genome. Analysis of mRNAs from virus-infected cells demonstrated that all three vaccinia virus serpin genes are transcribed early during infection. The amino acid sequences at the active sites of these serpins suggest that they may inhibit serine proteinases of differing biochemical specificities. The possible functions of these genes are discussed.
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The responses to slow constant velocity rotations in the clockwise (CW) and counterclockwise (CCW) directions about an axis tilted 10 degrees from the earth's vertical were studied in static tilt-sensitive neurons in the vestibular nuclei of decerebrate cats. Each unit responded to any 360 degrees unidirectional rotation with a position-dependent discharge maximum. The location of the maximum, obtained by rotation in one direction, differed from that obtained by an oppositely directed rotation (phase difference). In about 80% of the units such phase difference (up to 160 degrees in second-order neurons) in response to oppositely directed rotations was unaffected by different amplitudes of head displacement (5-25 degrees). Units were thus classified into two groups depending on the location of the CW discharge maximum relative to the CCW counterpart. The direction of rotation had no influence on the response gains of these units.