Gene expression of FK 506-binding protein.
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Amino Acid Isomerases↗
Biomedical subjects
S Kar
Publications and source records attributed to S Kar.
Origins and projections of peptide-immunoreactive nerves in the male rat genitofemoral nerve.
Following injection of retrograde tracer to one genitofemoral nerve of male rats, motoneurones and dorsal root ganglion cells of segmental levels L1/L2 were labelled ipsilaterally. Many labelled motoneurones were calcitonin gene-related peptide- (CGRP) immunoreactive. In the ganglia, a proportion of the labelled cells were CGRP-, tachykinin- or galanin-immunoreactive (10%:6%:53% respectively). In comparison with ganglia of the contralateral side, galanin-immunoreactive cells were significantly increased in the ipsilateral ganglia. Unilateral genitofemoral nerve section induced a loss of CGRP- or tachykinin- and an increase of galanin-immunoreactive cells in the ipsilateral L1/L2 ganglia. In the ipsilateral L1/L2 dorsal horn, CGRP and tachykinin, but not galanin, immunoreactivity was reduced. In the cremaster muscle and scrotal skin of either side galanin-immunoreactive fibres were not visible and CGRP- and tachykinin-immunoreactive fibres were depleted in the ipsilateral side. Capsaicin induced a loss of CGRP- and tachykinin-immunoreactive ganglion cells and of CGRP, tachykinin or galanin immunoreactivity from the dorsal horn. In the scrotal skin, CGRP- and tachykinin-immunoreactive fibres were depleted. By contrast in the muscle, a few CGRP-immunoreactive fibres persisted. The data demonstrate that (i) the genitofemoral nerve originates at segmental levels L1/L2; (ii) CGRP- and tachykinin-immunoreactive sensory and CGRP-immunoreactive motor neurones project to the cremaster muscle and scrotal skin; and (iii) nerve pertubation induces an increase of galanin-immunoreactive sensory neurones, the significance of which remains to be elucidated.
Animals↗
Time-related decrease of substance P and CGRP in central and peripheral projections of sensory neurones in Mycobacterium leprae infected nude mice: a model for lepromatous leprosy in man.
We have previously shown the depletion of cutaneous calcitonin gene-related peptide (CGRP)- and substance P-containing nerves in human leprosy. The aims of this study were to investigate the temporal effects of leprosy on nerves in skin and spinal cord. Tissues were taken from nude mice, 6 and 12 months after inoculation of Mycobacterium leprae into the hind footpads, and from age-matched controls. Sections were immunostained with antisera to substance P or CGRP. After 6 months of infection, substance P- and CGRP-immunoreactive nerves were reduced in skin from all body areas; by 12 months, the reduction was substantially greater. In the spinal cord, sensory fibres immunoreactive for substance P had decreased compared with controls at 6 and 12 months [by 60 per cent (0.022 mm2) and 80 per cent (0.048 mm2), respectively, P less than 0.001], as with CGRP [30 per cent (0.018 mm2) (P less than 0.02) and 40 per cent (0.028 mm2) (P less than 0.01), respectively]. CGRP immunoreactivity was completely absent in motor neurones after 12 months of infection. Loss of CGRP- and substance P-immunoreactive fibres in skin and spinal cord, and CGRP in motor neurones is in accord with impaired pain sensation and muscle weakness in leprosy.
Animals↗
The immunocytochemical distribution of seven peptides in the spinal cord and dorsal root ganglia of horse and pig.
The distribution of calcitonin gene-related peptide (CGRP), enkephalin, galanin, neuropeptide Y (NPY), somatostatin, tachykinins and vasoactive intestinal polypeptide (VIP) was compared in cervical, thoracic, lumbar and sacral segmental levels of spinal cord and dorsal root ganglia of horse and pig. In both species, immunoreactivity for the peptides under study was observed at all segmental levels of the spinal cord. Peptide-immunoreactive fibres were generally concentrated in laminae I-III, the region around the central canal, and in the autonomic nuclei. A general increase in the number of immunoreactive nerve fibres was noted in the lumbosacral segments of the spinal cord, which was particularly exaggerated in the case of VIP immunoreactivity. In the horse, some CGRP-, somatostatin- or tachykinin-immunoreactive cell bodies were present in the dorsal horn. In the pig, cells immunoreactive for somatostatin, enkephalin or NPY were noted in a similar location. In the ventral horn most motoneurones were CGRP-immunoreactive in both species. However, in pig many other cell types were CGRP-immunoreactive not only in the ventral horn, but also in laminae V-VI of the dorsal horn. With the exception of enkephalin and NPY immunoreactivity, which was not seen in pig dorsal root ganglia, all peptides studied were localised to neuronal cell bodies and/or fibres in the dorsal root ganglia. In both species, immunolabeled cell bodies were observed in ganglia from cervical, thoracic, lumbar and sacral levels, with the exception of VIP-immunoreactive cells that were detected only in the lumbosacral ganglia. Numerous CGRP- and tachykinin-immunoreactive cell bodies were visualised in both species, while the cells immunolabeled with other peptide antisera were much lower in number. In both species, immunostaining of serial sections revealed that a subset of CGRP-immunoreactive cells co-expressed tachykinin, galanin or somatostatin immunoreactivity. In the horse some enkephalin-immunoreactive cells were also CGRP positive and occasionally combinations of three peptides, e.g. CGRP, tachykinin and galanin or CGRP, tachykinin and enkephalin were identified. The results obtained suggest that the overall pattern of distribution of peptide immunoreactivities is in general agreement with that so far described in other mammals, although some species variations have been observed, particularly regarding the presence of immunoreactive cell bodies in the dorsal horn of the spinal cord.
Animals↗
Reversal of chronic regional myocardial dysfunction (hibernating myocardium) by synchronized diastolic coronary venous retroperfusion during coronary angioplasty.
A 62 year old man with previous myocardial infarction, an occluded right coronary artery and a 90% stenosis of the left anterior descending coronary artery underwent angioplasty with the support of coronary venous retroperfusion of arterial blood during the procedure. In two of four angioplasty balloon dilations of the left anterior descending coronary artery, synchronized diastolic retroperfusion of the coronary veins with arterial blood was applied to protect the severely dysfunctioning myocardium from additional ischemia. Two-dimensional echocardiography was used to monitor and quantitate alterations in left ventricular function. Retroperfusion of arterial blood resulted in immediate improvement in ischemic zone wall motion despite the totally occluded artery during balloon dilation. Echocardiographic images recorded after angioplasty showed a marked improvement in contraction of the previously dyskinetic segments, with changes similar to those seen during balloon dilations with synchronized diastolic coronary venous retroperfusion. Thus, in this patient, viability of chronically dysfunctioning myocardium could be demonstrated by the improvement in regional wall motion during retroperfusion. This technique could eventually be of value to elucidate the anatomic location of viable myocardium while maintaining adequate left ventricular systolic function during coronary artery interventions in the catheterization laboratory.
Angioplasty, Balloon, Coronary↗
Pronounced accumulation of metoprolol in ischemic myocardium after coronary venous retroinfusion.
The myocardial availability of the beta 1-selective blocker metoprolol was compared following standard intravenous (i.v.) administration and after coronary venous retroinfusion. Thirteen open-chest farm pigs were subjected to 90-min occlusion of the left anterior descending coronary artery. In six of these pigs, metoprolol was administered as an i.v. injection while 7 pigs received the drug retrogradely into the coronary vein. The time of administration was 5 min. In both groups, metoprolol was administered after 30 min of coronary artery occlusion. Metoprolol did not influence heart rate (HR) or blood pressure (BP) whether administered i.v. or into the coronary vein. At the end of administration, plasma metoprolol was significantly higher when administered i.v. (2,955 +/- 543 nmol/L) than after coronary venous infusion (1,213 +/- 464 nmol/L; p less than 0.05). At 30 and 60 min after injection, plasma metoprolol did not differ significantly between the two groups. Myocardial tissue concentration of metoprolol in nonischemic myocardium was approximately 480 pmol/g for both groups and similar in the subendocardial, midmyocardial, and subepicardial layers of the myocardium. After i.v. administration, myocardial Metoprolol concentration in the ischemic zone was less than that in the nonischemic zone, averaging 150-300 pmol/g tissue. In contrast, coronary venous retroinfusion of metoprolol resulted in a substantial accumulation of the drug in the ischemic zone, as exemplified by a subendocardial concentration of 2,002 +/- 689; a midmyocardial concentration of 26,643 +/- 8,813 and a subepicardial concentration of 98,571 +/- 58,930 pmol/g, respectively (mean +/- SE). Coronary venous retroinfusion of metoprolol resulted in a pronounced accumulation of drug in the ischemic myocardium.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Cloning and expression in Escherichia coli of LKP pilus genes from a nontypeable Haemophilus influenzae strain.
Nontypeable Haemophilus influenzae HF0295, isolated by aspiration from the middle ear of a patient with otitis media, expresses long, thick, and hemagglutinating pili of a single serotype (LKP1) on its surface. An intact pilus vaccine consisting of the LKP1 serotype protected chinchillas against experimental otitis media (C. C. Brinton, Jr., M. J. Carter, D. B. Derber, S. Kar, J. A. Kramarik, A. C. C. To, S. C. M. To, and S. W. Wood, Pediatr. Infect. Dis. J. 8:554-561, 1989; R. B. Karasic, D. J. Beste, S. C. M. To, W. J. Doyle, S. W. Wood, M. J. Carter, A. C. C. To, K. Tanpowpong, C. D. Bluestone, and C. C. Brinton, Jr., Pediatr. Infect. Dis. J. 8:562-565, 1989). The genes encoding LKP1 pili were cloned from a genomic library of the clinical strain as a 12.5-kilobase insert on a plasmid vector and inserted into Escherichia coli K-12. Transposon mutagenesis and deletion constructs mapped the pilus-coding region within a 7-kilobase region of insert DNA. The recombinant bacteria were found by electron microscopy to express pili morphologically similar to LKP1 pili. Purified pilus rods from the recombinant and its parental strain were composed of a single detectable protein with an apparent molecular weight of 27,500. Antibodies raised against LKP1 pili purified from H. influenzae immunologically reacted with pili from the recombinant bacteria. Pili from both strains also adhered to human erythrocytes and buccal cells with the same specificity.
Bacterial Adhesion↗
Ischemic brain rescue by transvenous perfusion in baboons with venous sinus occlusion.
We studied brain retroperfusion in nine adult baboons. Experiments in four baboons determined techniques and the safety of retroperfusion, and experiments in three baboons determined the ability of retroperfusion to reverse cerebral ischemia. Two baboons died before retroperfusion. Arterial blood was continuously circulated by an external pumping system from one femoral artery into the intracranial sinuses through specially designed balloon-tipped catheters placed percutaneously into the sigmoid sinuses bilaterally. The balloons intermittently occluded the sinuses. Ischemia was produced by occluding the left middle cerebral artery. Standard and computed electroencephalography with topographic mapping monitored the onset and reversal of ischemia. Retroperfusion rate exceeded 50 ml/min with a mean intrasinus pressure increase of 27 (0-149) mm Hg in all seven experiments. Venograms demonstrated complete or partial filling of the superior sagittal sinus in each experiment. Four experiments without ischemia established maximal balloon occlusion cycles, retroperfusion rates, and sinus pressure changes. These four baboons were neurologically normal after retroperfusion; two had normal magnetic resonance imaging scans. Ischemic changes, detected by electroencephalography following middle cerebral artery occlusion, were reversed with retroperfusion in all three ischemia experiments. Autopsies in the seven baboons demonstrated no parenchymal hemorrhage or edema. Our results suggest that further investigation of retroperfusion, and possibly retroinfusion of agents for cerebral protection, is warranted.
Animals↗
Antitoxoplasma antibody in clinically suspected cases of human toxoplasmosis.
(1) Seventy sera from a variety of patients suffering from different diseases suspected to be caused by Toxoplasma gondii infection and forty from non-toxoplasmic hospital cases and laboratory and hospital staff were collected. (2) Antitoxoplasma antibody was detected in those sera by Indirect Haemagglutination test (IHA test) and Enzyme Linked Immunosorbent Assay (ELISA). (3) Twenty one (30%) sera out of 70 test samples and 2 (5%) out of 40 control samples were positive by ELISA test. With IHA test only 17 (24.3%) of test samples and same 2 (5%) of control samples were positive. (4) Sera collected from Paediatric Department showed the highest positivity (40%) followed by Opthalmological group (35.7%) and obstetrics and Gynaecological group (13.6%). (5) No significant co-relation was found between the seropositivity with sex, diet and history of cat contact of the patients.
Animals↗
Novel peptide pancreastatin: its occurrence and codistribution with chromogranin A in the central nervous system of the pig.
The distribution of pancreastatin immunoreactivity was investigated in porcine brain, spinal cord, dorsal root ganglia, and pituitary. In the brain, immunoreactive cell bodies were present in many areas including the cortex, basal ganglia, hippocampus, thalamus, hypothalamus, mesencephalic reticular formation, cerebellum, and medulla oblongata. Immunoreactive fibres were most abundant in the globus pallidus, stria terminalis, entopeduncular nucleus, hippocampus, and in the substantia nigra. In the spinal cord, immunoreactive cells were found in laminae IV-IX. Immunoreactive fibres were concentrated in the dorsal horn. Pancreastatin immunoreactivity was localised to fibres and small cells (5-10% of the total) in the dorsal root ganglia. In the posterior pituitary, many immunoreactive fibres were present and in the anterior lobe subsets of gonadotrophs and thyrotrophs were pancreastatin-immunoreactive. The localisation of pancreastatin showed a parallel distribution with chromogranin A. Coexistence of pancreastatin with calcitonin gene-related peptide (CGRP) immunoreactivity in cell bodies in the spinal cord, including motoneurones, and with CGRP or galanin immunoreactivities in dorsal root ganglion cells was also noted. The differential pattern of pancreastatin immunostaining was reflected in the extractable levels of peptide with highest concentrations in the cortex (55.8 +/- 6.0 pmol/g wet weight, mean +/- S.E.M.), thalamus (60.0 +/- 5.0 pmol/g), hypothalamus (54.4 +/- 6.5 pmol/g), and anterior pituitary (2,714 +/- 380 pmol/g). Characterisation of pancreastatin immunoreactivity in the hypothalamus and pituitary by gel permeation and high-pressure liquid chromatography revealed multiple molecular forms, one of which was indistinguishable from natural porcine pancreastatin. The widespread distribution of pancreastatin immunoreactivity suggests this peptide may play a part in several neuroendocrine, autonomic, somatic, and sensory functions, and its colocalisation with chromogranin A is consistent with a precursor-product relationship.
Animals↗
Reduced numbers of calcitonin gene-related peptide-(CGRP-) and tachykinin-immunoreactive sensory neurones associated with greater enkephalin immunoreactivity in the dorsal horn of a mutant rat with hereditary sensory neuropathy.
The mutilated foot rat is a mutant with autosomal recessive sensory neuropathy and frequent mutilation of the hindlimbs. Decreased numbers of dorsal root ganglion cells and diminished sensitivity to painful stimuli are characteristics of these animals. By use of immunocytochemistry, changes in the distributions of peptides involved in sensory and/or autonomic regulation, i.e., calcitonin gene-related peptide (CGRP), tachykinins, enkephalin and neuropeptide Y in spinal cord, dorsal root ganglia and skin of these animals, were studied. In comparison with normal litter-mate controls, the dorsal horn of mutilated foot rats contained substantially fewer CGRP- and tachykinin-immunoreactive fibres but more fibres immunoreactive for enkephalin. Many enkephalin-immunoreactive cell bodies were also found in the dorsal horn of the mutants, by contrast none were visible in control animals. Neuropeptide Y immunoreactivity was, however, unchanged in the spinal cord of the mutants. In the dorsal root ganglia of the mutants, the number of CGRP- or tachykinin-immunoreactive cells and their proportion to total neuronal numbers were significantly less in comparison with normal controls. The diameter range of CGRP- and tachykinin-immunoreactive cells shifted from small (15-25 microns) to medium size (25-45 microns) as revealed by frequency distribution histograms. The skin from the affected fore- and hindlimbs of the mutant rats, in keeping with fewer CGRP- and tachykinin-immunoreactive cells in the dorsal root ganglia, contained substantially less fibres immunoreactive for CGRP and tachykinins; a difference that was not seen in skin of unaffected areas (whiskers and snout). By contrast, neuropeptide Y-immunoreactive fibres showed a normal distribution around blood vessels and sweat glands of mutilated foot rats. The data suggest that diminished pain perception in the mutilated foot rat is related to loss of peptide-containing sensory neurones. Furthermore, the intraspinal increase of enkephalinergic neurons in the dorsal horn, concomitant with the decreased number of primary sensory neurones, may also play a contributory rôle in reducing pain thresholds.
Animals↗
Design and development of pilus vaccines for Haemophilus influenzae diseases.
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Antibodies, Bacterial↗
Calcitonin gene-related peptide messenger RNA is expressed in sensory neurones of the dorsal root ganglia and also in spinal motoneurones in man and rat.
Calcitonin gene-related peptide (CGRP) mRNA was localised to neurones of the dorsal root ganglia and motoneurones of the ventral horn in man and rat. Presence of alpha- and beta-CGRP mRNA was confirmed by Northern blot analysis of rat tissues which showed alpha-CGRP was the predominant gene. The distribution of CGRP gene transcripts corresponded with neurones displaying CGRP immunoreactivity in the ganglia of both species and in the rat ventral horn. In man few motoneurones were immunoreactive despite many expressing CGRP mRNA. In situ hybridisation revealed not only sensory but also motor neurones are sites of CGRP manufacture. Thus in conjunction with other evidence the present study reinforces the proposed muscle trophic role for this peptide.
Aged↗
Echocardiographic criteria to distinguish reversible from irreversible myocardial ischaemia.
This treatise reviews two-dimensional echocardiographic criteria which have been developed to describe and distinguish reversible vs irreversible myocardial ischaemia. It also discusses the new pathophysiologic concepts such as 'hibernating' and 'decapitated' myocardium, and also 'reperfusion injury' and 'stunned' myocardium, complications which may supervene following reperfusion of jeopardized ischaemic myocardium. Computerized regional and global wall-motion analysis is now usually measured from enhanced endocardial edges. Provocative interventions can contribute information regarding viability of jeopardized ischaemic regions by testing contractile response of the myocardium to afterload reducing agents such as nitroglycerine or nitroprusside. They can also validate viability by demonstrating that post-extrasystolic beats can still cause potentiation. Ultrasonic contrast washout half-life of the myocardium which is compromised by stenotic coronary arteries provides a promising method for supplying information about the coronary perfusion defects and flow reserve. The decrease in global or regional ejection fraction following exercise echocardiography may show if jeopardized ischaemic myocardium is irreversibly damaged. A new hypercontractility phenomenon is described following brief coronary occlusions such as during percutaneous transluminal angioplasty, or after sudden release of angiospasm, and this should be considered a sign of viability. Increase in end-diastolic wall thickness and echo amplitudes immediately after reperfusion of ischaemic segments is often associated with reversibly damaged myocardium.
Coronary Disease↗
Ontogeny of the hypothalamo-neurohypophysial system in the toad, Bufo melanostictus: an immunohistochemical study.
Employing unlabeled antibody enzyme technique, the development of the hypothalamo-neurohypophysial system of the common Indian toad, Bufo melanostictus, has been studied by using antiserum raised in rabbit against bovine neurophysin. Immunoreactivity to the antiserum is first observed at stage 22 (Gosner, 1960) in the presumptive preoptic nucleus and at stage 23 in the presumptive median eminence and pars nervosa. As development proceeds, progressive increase in the immunoreactivity in all the components of the hypothalamo-neurohypophysial system is observed. The detection of neurohypophysial hormones (stages 22-23) much before the appearance of limb buds (stages 25-26) may be due to the greater sensitivity of the immunohistochemical technique.
Animals↗
Effects of intracoronary infusion of arterial blood or Fluosol-DA 20% on regional myocardial metabolism and function during brief coronary artery occlusions.
Effects of intracoronary infusion (50 ml/min) of arterial blood, oxygenated or unoxygenated Fluosol, or Plasmalyte A on hemodynamics, electrocardiogram, regional myocardial function, and lactate metabolism were studied in six closed-chest dogs during 2 min occlusions of the left anterior descending coronary artery followed by 10 min of reperfusion. Normal hemodynamics were maintained with infusion of arterial blood and oxygenated Fluosol, whereas unoxygenated Fluosol and Plasmalyte A resulted in hemodynamic deterioration similar to that noted with no treatment. Ischemic zone systolic fractional area change, an index of systolic function measured by two-dimensional echocardiography, remained normal during the occlusion supplemented with intracoronary arterial blood (49 +/- 7%), was moderately hypokinetic with oxygenated Fluosol (31 +/- 10%), and became severely hypokinetic with unoxygenated Fluosol (14 +/- 14%), with Plasmalyte A (2 +/- 13%), and in the absence of treatment (5 +/- 9%). Only infusion of arterial blood resulted in no ST segment elevation or lactate production. Thus intracoronary infusion of arterial blood during brief coronary occlusion maintained normal myocardial function and aerobic metabolism. Infusion of oxygenated Fluosol resulted in amelioration of the decline in regional function after coronary occlusion, but not complete protection.
Angioplasty, Balloon↗
Cytodifferentiation and immunocharacteristics of adenohypophysial cells in the toad, Bufo melanostictus.
Employing the unlabelled antibody enzyme technique cytodifferentiation, immunocharacteristics and topographical distribution of melanotropic (MSH), adrenocorticotropic (ACTH), thyrotropic (TSH), prolactin (PRL), gonadotropic (GTH) and growth hormone (GH) secreting cells in the embryonic/larval as well as adult pituitary gland of the common Indian toad, Bufo melanostictus, have been studied by using antisera raised in rabbit against mammalian hypophysial hormones. Immunoreactive MSH and ACTH cells appear first in the dorsocaudal and rostral regions of the pituitary anlage (PA) at stage 21 (Gosner's classification) of the embryonic development. This is followed by the differentiation of TSH and PRL cells at stage 22 in the midventral and central regions of the PA respectively. Finally, at stage 23 the GTH cells appear in the rostral and the GH cells in the caudal regions of the PA. With the progress of the development, cells showing immunoreactivity to various antisera gradually increase in number, size, granular content and finally occupy the characteristic adult disposition. The MSH cells comprise the pars intermedia. In the pars distalis (PD) the ACTH cells are localized in the rostroventral region, TSH cells in the central region and the GH cells in the dorsocaudal region. However, GTH and PRL cells are distributed throughout the PD.
Adrenocorticotropic Hormone↗
Entry of Bartonella bacilliformis into erythrocytes.
Bartonella bacilliformis, which causes the human diseases Oroya fever and verruga peruana, binds to human erythrocytes in vitro and produces substantial and long-lasting deformations in erythrocyte membranes, including cone-shaped depressions, trenches, and deep invaginations. The deforming force is probably provided by the polar flagella of these highly motile bacteria. Deep invaginations containing bacteria are commonly seen, and membrane fusion at the necks of the invaginations leads to the formation of intracellular vacuoles containing bacteria. Fluorescent compounds present externally render the vacuoles fluorescent and, occasionally, lightly fluorescent cells are seen, suggesting that the vacuoles sometimes rupture to admit the bacteria to the cytoplasm. Vacuoles present in fluorescent erythrocytes prepared by preloading the erythrocytes with fluorescent compounds are seen as dark areas from which the fluorescent marker is excluded. Entry of the bacteria appears to be the result of a process of forced endocytosis.
Bacterial Adhesion↗