1. Neuropeptide-induced histamine release is thought to occur via receptor-independent mechanisms, with net charge and lipophilicity being important factors. 2. In this study, the histamine releasing ability of neuropeptide Y (NPY), two C-terminal segments of NPY and 13 centrally truncated NPY analogues was examined. These results were compared with the ability of the peptides to bind to the Y2 receptor in the rabbit kidney membrane model and with their hypotensive actions in the anaesthetized-rat model. 3. All analogues tested, with the exception of [Glu4,25,33,35]-NPY(1-4)-Ahx-(25-36) and [Asp4,25,33,35]NPY(1-4)-Ahx-(25-36) which were devoid of histamine releasing activity, evoked a dose-dependent histamine release but there were marked differences between the peptides. The native peptide was the least active. 4. Histamine release was not linked to the ability of the peptides to displace NPY from Y2 receptors. There was a statistical correlation between the hypotensive effects expressed as ED10 values (mumol kg-1, which induced a blood pressure decrease of 10 mmHg) and the EC25 for histamine release (r = 0.62, P = 0.04), although histamine release may not be the sole determinant of the alterations in blood pressure. 5. There was a strong negative correlation between EC25 for histamine release and net positive charge (r = -0.93, P = 5.7 x 10(-7), i.e. increasing the net positive charge caused greater histamine release. However, there was a 12 fold difference in activity amongst the most positively charged analogues (+5). Helicity did not correlate with histamine releasing ability. 6. In the development of NPY-related drugs the avoidance of compounds with net positive charge is recommended.

Animals↗

PubMed1996

Medial preoptic area afferents to periaqueductal gray medullo-output neurons: a combined Fos and tract tracing study.

We have shown recently that the medial preoptic area (MPO) robustly innervates discrete columns along the rostrocaudal axis of the midbrain periaqueductal gray (PAG). However, the location of PAG neurons responsive to MPO activation is not known. Anterograde tract tracing was used in combination with Fos immunohistochemistry to characterize the MPO --> PAG pathway anatomically and functionally within the same animal. Focal electrical or chemical stimulation of MPO in anesthetized rats induced extensive Fos expression within the PAG compared with sham controls. Fos-positive neurons were organized as 2-3 longitudinal columns. The organization and location of these columns overlapped remarkably well with the distribution of fibers and terminals in PAG labeled by Phaseolus vulgaris leucoagglutinin (PHA-L) injected into the same MPO stimulation site. This indicates that MPO inputs may terminate on the soma or proximal dendrites of neurons exhibiting elevated Fos. A second series of experiments investigated whether MPO stimulation excited PAG neurons with descending projections to the medulla. Retrograde labelling of PAG neurons projecting to the medial and lateral regions of the rostroventral medulla (RVM) was combined with MPO-induced Fos expression. The results showed that a substantial population (37-53%) of Fos-positive PAG neurons projected to the ventral medulla. This indicates that MPO stimulation engages PAG-medullary output neurons. Taken together, these results suggest that the MPO --> Pag --> RVM projection constitutes a functional pathway. This circuit may coordinately regulate neuroendocrine, motor, and autonomic adjustments necessary for the elaboration of sexual behaviors.

Afferent Pathways↗

PubMed1996

Investigating paediatric airways by non-bronchoscopic lavage: normal cellular data.

BACKGROUND: Bronchoscopic bronchoalveolar lavage in children to investigate bronchial disorders such as asthma has both ethical and procedural difficulties. OBJECTIVE: The aim of this study was to establish a standardized non-bronchoscopic method to perform bronchoalveolar lavage in children attending for elective surgery to obtain normal cellular data. METHODS: Bronchoalveolar lavage was performed on normal children (n = 55) by infusing saline (20 mL) through an 8 FG suction catheter passed after endotracheal intubation. Oxygen saturation, heart and respiratory rate were monitored during the bronchoalveolar lavage procedure. Cellular analysis and total protein estimation of the lavage fluid were performed. Epithelial lining fluid volume was calculated (n = 15) using the urea dilution method. RESULTS: The procedure was well tolerated by all children. Total cell count and differential cell count for children (macrophages 70.8 +/- 2.3%, lymphocytes 3.8 +/- 0.6%, neutrophils 5.7 +/- 1.0%, eosinophils 0.14 +/- 0.03%, epithelial cells 19.6 +/- 2.1%, mast cells 0.21 +/- 0.02%) were similar to those reported for adults. Age and sex comparisons revealed no differences between groups. The mean total protein recovered in the cell free supernatant was 49.72 +/- 4.29 mg/L and epithelial lining fluid volume was 0.82 +/- 0.11% of return lavageate. CONCLUSION: This method allows bronchoalveolar lavage to be performed safely and quickly on children attending for routine elective surgery. Using this method and taking the 'window of opportunity' of elective surgery, the presence or absence of airway inflammation could be studied in children with various patterns of asthma during relatively asymptomatic periods.

Adolescent↗

PubMed1995

Influence of alpha-helicity, amphipathicity and D-amino acid incorporation on the peptide-induced mast cell activation.

Mast cell activation by polycationic substances is believed to result from a direct activation of G protein alpha subunits and it was suggested that the adaption of amphipathic, alpha-helical conformations would allow the peptide to reach the cytosolic compartment to interact with G proteins (Mousli et al., 194, Immunopharmacology 27, 1, for review). We investigated the histamine-releasing activity of model peptides as well as analogues of magainin 2 amide and neuropeptide Y with different amphipathicities and alpha-helix content on rat peritoneal mast cells. Amphipathic helicity is not a prerequisite for mast cell activation. Moreover, non-helical magainin peptides with high histamine-releasing activity were less active in the liberation of carboxyfluoresceine from negatively charged liposomes, indicating that peptide-induced mast cell activation and peptide-induced membrane perturbation do not correlate. In contrast to the negligible influence of the secondary structure, amino acid configuration may exert a striking influence on peptide-induced mast cell activation. Thus histamine-release by substance P was markedly impaired when the L-amino acids in the positively charged N-terminal region were replaced by D-amino acids, with [D-Arg1)substance P being the most inactive substance P diastreoisomer.

Amino Acid Sequence↗

PubMed1995

Fos expression induced by changes in arterial pressure is localized in distinct, longitudinally organized columns of neurons in the rat midbrain periaqueductal gray.

The distribution of neurons expressing Fos within the periaqueductal gray (PAG) following pharmacologically induced high or low blood pressure was examined to determine (1) if PAG neurons are responsive to changes in arterial pressure (AP) and (2) the relationship of these cells to the functionally defined hypertensive and hypotensive columns in PAG. Changes in AP differentially induced robust Fos expression in neurons confined to discrete, longitudinally organized columns within PAG. Increased AP produced extensive Fos-like immunoreactivity within the lateral PAG, beginning at the level of the oculomotor nucleus. At the level of the dorsal raphe, Fos expression induced by increased AP shifted dorsally, into the dorsolateral division of PAG; this pattern of Fos labeling was maintained throughout the caudal one-third of PAG. Double-labeling for Fos and nicotinamide adenine dinucleotide phosphate diaphorase confirmed that Fos-positive cells induced by increased AP were located in the dorsolateral division of PAG at these caudal levels. Fos positive cells were codistributed, but not colocalized, with nicotinamide adenine dinucleotide phosphate diaphorase-positive cells. Decreased AP evoked a completely different pattern of Fos expression. Fos-positive cells were predominantly located within the ventrolateral PAG region, extending from the level of the trochlear nucleus through the level of the caudal dorsal raphe. Double-labeling studies for Fos and serotonin indicated that only 1-2 double-labeled cells per section were present. Saline infusion resulted in very few Fos-like immunoreactive cells, indicating that volume receptor activation does not account for Fos expression in PAG evoked by changes in AP. These results indicate that (1) substantial numbers of PAG neurons are excited by pharmacologically induced changes in AP and (2) excitatory barosensitive PAG neurons are anatomically segregated based on their responsiveness to a specific directional change in AP.

Animals↗

PubMed1995

Neural anatomy of the glenohumeral ligaments, labrum, and subacromial bursa.

The neural histology of the human shoulder ligaments, glenoid labrum, and subacromial bursae were studied using a modified gold chloride stain. Two morphological types of mechanoreceptors and free nerve endings were found in the ligaments. Slow adapting Ruffini end organs and rapidly adapting Pacinian corpuscles were identified in the superior, middle, inferior, and the posterior glenohumeral ligaments. These specialized proprioceptive nerve endings were also found in the coracoclavicular, and coracoacromial ligaments. Only free nerve endings were found in the glenoid labrum and these were located in the peripheral half. Scattered free nerve endings were found throughout the subacromial bursae. This is the first histological evidence of neural receptors in the human shoulder ligaments, glenoid labrum, and the subacromial bursae. Any disruption of the labrum or these ligaments by trauma or surgery can deprive the shoulder of mechanical stability, and may cause a decrease in proprioception because of the loss of these afferent neural receptors. Removal of symptomatic, inflamed bursae may decrease pain signals from this area of the shoulder.

Adult↗

PubMed1995

A comparison of the determinants of safe injecting and condom use among injecting drug users.

A sample of 582 injecting drug users were interviewed as part of an evaluation of an AIDS prevention programme for drug users. This paper examines the biographic and predispositional determinants of five HIV preventive behaviours--equipment sharing (not receiving and not giving) and and condom use (with regular partners, casual partners and sex clients). A two-stage sequential approach was adopted for a logistic regression analysis. Initially, to model each of the five preventive behaviours, biographical and drug use variables were entered. In a second set of models, behavioural predisposition factors were included. Age, drug use and prison experience correlate with variables in both models, although not consistently in the same direction. While a predisposition to reject sharing correlates with safer rejecting and condom use, the predisposition to safer sex only correlates with condom use. Needle exchange programmes that only target the individual would seem to be inadequate. To enhance targeted interventions changes in public and agency policy that create a social environment conductive to behaviour change are required.

Acquired Immunodeficiency Syndrome↗

PubMed1995

A model for improving generalist physician output: the osteopathic experience.

Osteopathic medicine is identified closely with primary care. Approximately one-half of all doctors of osteopathy (DOs) are family practitioners. The educational model responsible for producing such a high percentage of generalist physicians should be better understood for its achievements. Colleges of osteopathic medicine are unique in sharing a common mission of producing primary care physicians, and their methodologies reflect a similarly shared educational emphasis. Compared to allopathic medical schools, colleges of osteopathic medicine are the highest producers of generalist physicians because of several common features. This article evaluates colleges of osteopathic medicine by profiling each institution according to characteristics that have a potential impact on the output of generalist physicians: (1) institutional ownership; (2) age; (3) class size; (4) leadership; (5) educational focus; (6) whole-time as opposed to adjunct clinical faculty; (7) clinical educational settings; (8) clinical training sites; and (9) reliance on MD rather than DO clinical faculty. Comparisons between colleges of osteopathic medicine according to these characteristics yield mixed results but also highlight many differences from allopathic educational models. Factors separating the highest from lowest producers of generalist physicians vary at colleges of osteopathic medicine, and other characteristics or circumstances beyond their control may affect the number of graduates pursuing careers in primary care.

Career Choice↗

PubMed1995

Modulatory action of Helicobacter pylori on histamine release from mast cells and basophils in vitro.

Helicobacter pylori is important in the aetiology of peptic ulceration. Despite inducing an inflammatory response in the mucosa, the organism persists, suggesting that it has efficient protective mechanisms. Some bacterial and viral products modulate histamine secretion from inflammatory cells. Therefore, this study examined the modulatory effects of H. pylori preparations on histamine release from rat peritoneal mast cells and human basophils. Eleven clinical isolates of H. pylori were prepared in different ways: as whole washed bacteria, washed sonicated bacteria, and formalin-killed bacteria, and as outermembrane and lipopolysaccharide (LPS) extracts. Histamine release from mast cells or basophils was not elicited by any of these bacterial preparations alone. However, when mixed with various secretory stimulants, the bacterial preparations caused inhibition of histamine release from rat mast cells (calcium ionophore A23187, compound 48/80, concanavalin A, anti-rat IgE) and human basophils (A23187, N-formyl Met-Leu-Phe). The degree of inhibition ranged from 48% to 97%. These results indicate that H. pylori exerts an inhibitory effect on cells of the immune system that contributes to its persistence within the gastric mucosa.

Animals↗

PubMed1995

Substance P induces histamine release from human pulmonary mast cells.

Substance P elicits histamine release from human skin and rodent mast cells. Since neuropeptide-mediated reflexes may be important in asthma, we examined the ability of substance P to stimulate human mast cells obtained at bronchoalveolar lavage (BAL). BAL samples were obtained at routine bronchoscopy from 35 non-preselected patients. Histamine release experiments were performed in a standard manner using substance P and the calcium ionophore A23187. Both substance P (50 microM) and A23187 caused histamine release (median 26.7% range 6.2-62.8% and 32.1%, 7.7-56.8% respectively) which was significantly greater (P < 0.0001) than the spontaneous release (median 15.6%, range 4.1-33.4%), i.e. that in the absence of any stimulus. Substance P induced histamine release was via an energy dependent process and was blocked by preincubation with antimycin A. A significant correlation was observed between substance P induced release and spontaneous release but was not observed with A23187 induced release. Mast cell counts correlated significantly with substance P induced release but not with spontaneous or A23187 induced release. The substance P induced histamine secretion was elicited at similar concentrations to those used with rodent and human skin mast cells. Asthma is associated with increased numbers of mast cells which have both increased spontaneous and stimulated secretory responses. Thus, in vivo, the bronchoconstrictor action of substance P may in part result from activation of mast cells in the bronchial lumen.

Calcimycin↗

PubMed1995

Effect of in vivo and in vitro lovastatin treatment on mast cell activation.

The hydroxymethylglutaryl coenzyme A (HMG CoA) reductase inhibitor lovastatin is used to treat hyperlipidaemia. This agent prevents the isoprenylation of some proteins involved in signal transduction processes and inhibits IgE-receptor-linked mediator release from RBL-2H3 cells. In this study the effect of in vivo and in vitro administration of lovastatin on histamine release from rat peritoneal mast cells was examined. Lovastatin (4 mg/kg/day for 2 weeks) inhibited histamine release induced by concanavalin A (con A) from rat peritoneal mast cells of Hooded-Lister rats and both homozygous lean and obese Zucker rats. In contrast, release induced by antirat IgE (anti-IgE) was only significantly inhibited in cells derived from Hooded-Lister rats and that induced by compound 48/80 was not altered. Lovastatin (20 microM, 24 h, in vitro) caused a significant inhibition of the subsequent histamine release to con A, anti-IgE and compound 48/80 but not to the calcium ionophore A 23187. It is important to determine whether such inhibitory effects are also observed after the chronic, clinical administration of lovastatin and other HMG CoA reductase inhibitors.

Animals↗

PubMed1994

Directionally specific changes in arterial pressure induce differential patterns of fos expression in discrete areas of the rat brainstem: a double-labeling study for Fos and catecholamines.

Although the nucleus tractus solitarii (NTS) has been established as the primary site of synaptic integration for the baroreceptor reflex, the higher-order pathways responsive to, and mediating, changes in vasomotor tone are not well characterized. We used immunohistochemistry to determine the distribution of cells expressing the Fos protein following pharmacologically induced, directionally specific changes in arterial pressure. The goal of this investigation was to determine if this immediate early gene product is differentially expressed in neurons of the rat brainstem following increased (pressor) versus decreased (depressor) arterial blood pressure (AP). Because brainstem catecholaminergic (CA) cell groups have been implicated in cardiovascular regulation, a double-labeling immunohistochemical procedure was used to examine the distribution of Fos in CA cells. Animals received continuous intravenous infusion of either a vasoconstrictor (l-phenylephrine hydrochloride), a vasodilator (sodium nitroprusside), or physiological saline. Extensive Fos-like immunoreactivity (FLI) was induced in both the pressor and depressor conditions in the NTS, caudal ventrolateral medulla (CVLM), rostral ventrolateral medulla (RVLM), A5, locus coeruleus (LC), Kolliker-Fuse, and parabrachial nucleus (PBN). These regions have all been implicated in central cardiovascular regulation. There were differences in the anatomical distribution of Fos-positive cells along the rostrocaudal axis of CVLM in the pressor and depressor conditions. Specifically, increased AP induced significantly more FLI cells within the rostral aspects of CVLM, whereas decreased AP resulted in a significantly greater number of FLI cells within the caudal CVLM. This result suggests that selective vasomotor responses differentially engaged discrete subsets of neurons within this brainstem region. Overall, approximately 50% of CA-immunoreactive cells were also FLI (CA-FLI) in the A1, A5, and A7 regions. Interestingly, increased AP produced significantly more CA-FLI double-labeled cells within the caudal than rostral A1 compared with depressor and control groups. Additionally, increased AP yielded significantly less CA-FLI double-labeled cells within the caudal A2 region. This suggests that CA barosensitive neurons in the CVLM/A1 and NTS/A2 regions are functionally segregated along the rostrocaudal axis of these structures. While twice as many PNMT-FLI double-labeled neurons were found in the C1-C3 regions following vasomotor changes versus saline control, there were no differences in the numbers or anatomical locations of labeled cells between pressor versus depressor groups. The results of this study indicate that (1) tonic changes in AP induce robust Fos expression in brainstem cardiovascular areas and (2) neurons responsive to specific directional changes in arterial pressure are segregated in some brainstem regions.

Animals↗

PubMed1994

Preoptic projections to Barrington's nucleus and the pericoerulear region: architecture and terminal organization.

The medial preoptic area (MPO), a sexually dimorphic region, plays a pivotal role in neuroendocrine function and reproductive behavior. We recently reported that MPO projects heavily to the midbrain periaqueductal gray (PAG). We also noted that MPO projects to the dorsolateral pontine tegmentum. Here we identified the cells of origin of the MPO-->tegmental projection and delineated the terminal organization of MPO projections to Barrington's nucleus, locus coeruleus (LC), and the rostromedial pericoerulear region (pLCrm). Correlative cyto- and chemoarchitectonic studies were done to define better the nuclear groups of the dorsolateral pontine tegmentum. Retrograde tracing revealed that MPO neurons projecting to the dorsolateral pontine tegmentum are preferentially distributed in distinct subregions of MPO, including the sexually dimorphic medial preoptic nucleus (MPN). Anterograde tracing with wheat germ agglutinin-horseradish peroxidase or Phaseolus vulgaris leucoagglutinin demonstrated considerable target specificity in projections from MPO to the dorsolateral pontine tegmentum. Barrington's nucleus receives a dense focal input along its entire rostrocaudal axis. In addition, pLCrm is heavily targeted by MPO inputs; pLCrm contains a concentrated plexus of extranuclear dendrites of LC neurons. The lateral dorsal tegmental (LDT) nucleus and LC proper receive only sparse input from MPO. MPO projections to Barrington's nucleus could regulate micturition reflexes during reproductive behavior. The MPO-->pLCrm projection could influence noradrenergic LC neurons in relation to reproductive and/or gonadal steroid function. Given the strong established connections from olfactory structures to MPO, it is possible that the MPO-->LC pathway provides an anatomical substrate for olfactory modulation of arousal.

Animals↗