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D R Springall

Publications and source records attributed to D R Springall.

126 records · Page 7Linked to original sources
PubMed1987

Calcitonin gene-related peptide is localised to human airway nerves and potently constricts human airway smooth muscle.

In human airways synthetic human sequence calcitonin gene-related peptide (hCGRP), a novel peptide produced by alternative processing of mRNA from the calcitonin gene, caused concentration-dependent contraction of human bronchi (EC50 4.9 X 10(-9) M) and was significantly more potent than substance P or carbachol. The contractile response was unaffected by atropine (2 X 10(-6) M), propranolol (10(-6) M), indomethacin (10(-5) M), tetrodotoxin (3 X 10(-6) M), chlorpheniramine (10(-4) M), cimetidine (10(-5) M), or FPL55712 (10(-4) M) suggesting a direct effect of CGRP on airways smooth muscle. CGRP was detected in human airways by radioimmunoassay with highest concentrations in cartilaginous airways. CGRP was localised by immunocytochemistry to both nerves and ganglia in human airways. CGRP, is a potent constrictor of human airways and may have important effects on airway function and be implicated in the pathogenesis of bronchial hyper-responsiveness and asthma.

Aged↗
PubMed1987

Regulatory peptides in the respiratory tract of Macaca fascicularis.

The quantitative distribution and localisation of seven regulatory peptides (vasoactive intestinal peptide (VIP), peptide histidine methionine (PHM), calcitonin gene related peptide (CGRP), galanin, substance P, neuropeptide tyrosine (Y), and bombesin like peptides) were determined by radioimmunoassay and immunocytochemistry in six different regions of the respiratory tract of the cynomolgus monkey, Macaca fascicularis. In general, peptide concentrations were higher in the airways than in lung tissue itself. VIP and PHM were found in greatest abundance and in equimolar concentrations. Concentrations of substance P, neuropeptide Y, and bombesin were substantially lower. Immunocytochemistry localised all the peptides to nerve fibres, whose density generally paralleled the tissue concentrations by radioimmunoassay except in the case of bombesin, which was not detected. VIP, PHM, and galanin were mostly associated with glands of trachea and bronchus and with blood vessels and smooth muscle; CGRP and substance P were found principally beneath airway epithelium and around smooth muscle fibres and blood vessels; neuropeptide Y was found around blood vessels and seromucous glands only. The pattern of peptide distribution in the Macaca fascicularis respiratory tract is similar to that previously reported in human postmortem material, suggesting that the cynomolgus monkey may be a useful model for examining the pathophysiological role of peptides in human respiratory disease.

Animals↗
PubMed1986

Endocrine differentiation of extra-pulmonary small cell carcinoma demonstrated by immunohistochemistry using antibodies to PGP 9.5, neuron-specific enolase and the C-flanking peptide of human pro-bombesin.

Several recent studies have confirmed the endocrine nature of small cell carcinoma of the lung. In extra-pulmonary sites, small cell 'undifferentiated' carcinomas have classical morphological features similar to their pulmonary counterpart. We therefore investigated, using immunocytochemistry, the possibility that the non-pulmonary neoplasms may also be endocrine in nature. Sections of 29 small cell carcinomas from oesophagus, stomach, larynx, colon and urinary bladder were immunostained using antisera to protein gene product 9.5 (PGP 9.5), neuron-specific enolase (NSE), cytokeratin, leucocyte common antigen and peptides including bombesin, the C-flanking peptide of human probombesin, adrenocorticotrophic hormone, neurotensin, calcitonin and pancreatic polypeptide. All the tumours showed immunoreactivity for at least one of the two general endocrine markers PGP 9.5 and NSE. Twenty-three of the 29 cases were immunoreactive for PGP 9.5, 27 for NSE. All were positive for cytokeratin and negative for leucocyte common antigen. Of the regulatory peptides, immunoreactivity was obtained with antisera to bombesin (one case), the C-flanking peptide of human pro-bombesin (14 cases), adrenocorticotrophic hormone (one case) and calcitonin (three cases). No PGP 9.5-, NSE- or peptide-like immunoreactivity was detected in 25 control tumours from similar sites, including lymphomas and poorly differentiated tumours. These results suggest that non-pulmonary small cell carcinoma has an endocrine character.

Adrenocorticotropic Hormone↗
PubMed1986

Occurrence, distribution and ontogeny of CGRP immunoreactivity in the rat lower respiratory tract: effect of capsaicin treatment and surgical denervations.

The occurrence and distribution of calcitonin gene-related peptide (CGRP) immunoreactivity in the rat respiratory tract were investigated by means of immunocytochemistry and radioimmunoassay using antibodies raised in rabbits to synthetic rat CGRP. Substantial amounts of CGRP immunoreactivity (range 5-37 pmol/g) were detected in all parts of the respiratory tract, the highest being in the stem bronchus. Gel filtration chromatography of extractable CGRP immunoreactivity revealed one single peak, eluting at the position of synthetic rat CGRP. CGRP immunoreactivity was localized both in mucosal endocrine cells and nerve fibres from the larynx down to the peripheral lung. CGRP-immunoreactive endocrine cells were found singly in trachea and stem bronchi and in groups in intrapulmonary airways. They appeared at a late stage of gestation (17 days), reached a maximum number near term and decreased after birth to maintain a population similar to that of the adult animals by postnatal day 21. Similarly, CGRP-immunoreactive nerve fibres were first identified by day 18 of the gestation period and reached the adult distribution by postnatal day 21. CGRP-immunoreactive nerve fibres were localized among smooth muscle, seromucous glands, beneath and within the epithelium of the airways and around blood vessels. CGRP was also found in sensory ganglia and in motor end plates of the larynx musculature. Neonatal pretreatment with capsaicin caused a marked reduction in CGRP immunoreactivity of nerve fibres in the respiratory tracts as well as a less marked decrease in the population of CGRP-containing endocrine cells of the lung. No change was seen in motor end plates immunostaining. Vagal ligation experiments revealed that CGRP-immunoreactive nerve fibres travelling in the vagus originate mainly from neurons located in the jugular ganglion. Infranodosal right vagal ligation induced a marked loss in CGRP-immunoreactive nerves of the trachea, and of the ipsilateral stem bronchus, but no changes were observed in peripheral lung. By contrast infranodosal left side vagal ligation caused a decrease in CGRP-immunoreactive nerves of the ipsilateral lung and bronchus without affecting the peptide content in the trachea. Left vagal ligation also induced a marked increase in both the intensity of staining and number of CGRP-immunoreactive endocrine cells in the lung. We conclude that CGRP immunoreactivity is localized in both nerve fibres and endocrine cells and is associated principally with the afferent (sensory) innervation of the respiratory tract.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1985

Antibodies to neurofilament protein and other brain proteins reveal the innervation of peripheral organs.

Monoclonal and polyclonal antibodies to neurofilament proteins, neuron-specific enolase, glial fibrillary acidic protein and S-100 have been used to demonstrate nerves, ganglion cells and the supportive glial system of the innervation of various organs. The female genitalia, the urinary tract, the respiratory system, the pancreas, the heart and the skin of several mammalian species, including rat, mouse, guinea pig, cat, pig, monkey and man were fixed in para-benzoquinone and portions of each organ were snap frozen. Serial or free-floating thick cryostat sections were stained using indirect immunofluorescence and peroxidase anti-peroxidase immunocytochemistry. In addition, the newly described and highly sensitive immunogold-silver staining technique was used on Bouin's-fixed and wax-embedded tissues. Antibodies to neurofilament proteins seemed to react with neuronal structures in all the species studied. Alternately stained serial sections showed a similar distribution of neurofilament proteins and neuron-specific enolase-containing nerves. Neuron-specific enolase staining had a diffuse appearance and was found to be highly variable, indicating that the neuron-specific enolase content might be related to the physiological state of the nerves and ganglion cells, whereas antibodies to neurofilament protein gave a consistently intense and very clear picture of the ganglion cells and nerve fibres. Antibodies to S-100 stained supportive elements of the peripheral nervous system in all tissues examined, whereas antibodies to glial fibrillary acidic protein were more selective.

Animals↗
PubMed1985

The immunogold-silver staining method. A powerful tool in histopathology.

Immunostaining of routinely fixed, wax embedded tissues may present problems to the pathologist since destruction of antigens can lead to false negative results. In an attempt to overcome this problem, we have compared the results of the standard peroxidase anti-peroxidase (PAP) method with those obtained using the newly developed and very sensitive immunogold-silver staining (IGSS) method. Sections from routine histopathological material as well as from normal tissue specimens were used in the comparison. Antisera to a variety of antigens commonly employed in pathology were used, including regulatory peptides and a range of other markers. In all cases the IGSS method was found to give superior or at least equal results to those obtained with the PAP technique. In some cases staining was obtained with IGSS method when the PAP technique gave no result. The intense black reaction product allowed much easier and more rapid screening of immunostained preparations as well as permitting sections to be counterstained with routine histological stains such as haematoxylin and eosin. It is therefore suggested that immunogold-silver staining is a valuable technique for the pathologist, particularly when examining overfixed or badly processed tissues.

Animals↗
PubMed1985

Gastrin-releasing peptide-like immunoreactivity in medullary thyroid carcinoma.

Gastrin-releasing peptide, the mammalian counterpart of amphibian bombesin, has been found to be present in high concentration by radioimmunoassay in eight histologically confirmed medullary thyroid carcinomas and to be undetectable in postmortem normal thyroid tissue. Chromatographic analysis of the tumor extracts by gel permeation revealed two major peaks of gastrin-releasing peptide-like immunoreactivity (GRP-LI). However, reverse-phase high-pressure liquid chromatography demonstrated three immunoreactive peaks of GRP-LI. None of these immunoreactive peaks was coeluted with synthetic porcine GRP or amphibian bombesin, but one of the peaks exactly emerged in the position of neuromedin C (C-terminal decapeptide of GRP). Sections from nine primary or secondary tumours were immunostained for GRP using a peroxidase/anti-peroxidase technic. All the medullary thyroid carcinomas were shown to contain GRP-LI, specifically localized to the tumor cells. This immunoreactivity is elevated in plasma from some patients with this malignancy, raising the possibility that it may be used as an additional tumor marker.

Carcinoma↗
PubMed1985

Distribution of galanin immunoreactivity in the respiratory tract of pig, guinea pig, rat, and dog.

Galanin, a newly discovered peptide isolated from porcine intestine, is known to cause contraction in rat smooth muscle preparations and to induce hyperglycaemia in dogs. By the use of radioimmunoassay and immunohistochemical techniques the concentration and distribution of galanin immunoreactivity were determined in several areas of the respiratory tract of five dogs, five guinea pigs, five rats, and two pigs. Antibodies were raised in rabbits to whole unconjugated natural porcine galanin. The highest galanin concentrations were found in the bronchus and the trachea of the dog, guinea pig, rat (2 pmol/g in each case), and pig (less than 1 pmol/g). The lowest galanin concentrations were found in the lung parenchyma. Gel chromatographic analysis in the pig showed one molecular form of galanin coeluting with the porcine galanin standard. By means of the indirect immunofluorescence technique on sections of tissues fixed in benzoquinone solution, galanin was found to be confined to nerve fibres in different regions of the respiratory tract. In the nasal mucosa of the pig nerve fibres containing galanin were distributed around seromucous glands and blood vessels and beneath the epithelium. In the trachea, bronchus, and major intrapulmonary airways of the pig, dog, and guinea pig galanin immunoreactive fibres were detected predominantly in smooth muscle, as well as around seromucous glands and in the adventitia of blood vessels. Rarely, galanin immunoreactive nerve fibres were found in the lung parenchyma. A few galanin immunoreactive ganglion cells also containing vasoactive intestinal polypeptide were found in the adventitia of the tracheobronchial wall of the pig and dog. The distribution of galanin suggests that it may have some influence on airway, vascular, and secretory functions in the mammalian respiratory tract.

Animals↗
PubMed1984

The immunocytochemical detection of herpes simplex virus in cervical smears--a valuable technique for routine use.

Herpes simplex virus type 2 is demonstrated in routine cervical smears by immunocytochemistry. Optimum antibody dilution and suitable fixation were determined and immunostaining results were found to compare favourably with those obtained by screening Papanicolaou-stained smears or by virus culture methods (virus isolation). Results obtained for immunostaining of smears from 158 patients agreed with those of virus isolation in 47 of 50 cases where virus isolation was performed. It is suggested that this immunocytochemical technique is valuable in detection of Herpes simplex virus in cervical smears, offering an inexpensive and rapid laboratory diagnosis with advantage to both patient and clinician.

Biopsy↗
PubMed1984

Immunostaining of neuron-specific enolase is a valuable aid to the cytological diagnosis of neuroendocrine tumours of the lung.

The neuroendocrine marker neuron-specific enolase (NSE) has been reported to be detectable by immunohistochemistry in paraffin sections of neuroendocrine neoplasms of the lung. There is no study of immunodetection of NSE in cytological smear preparations from these tumours. We have examined 10 cases of small cell carcinoma of the lung, using cells obtained from serous fluids or bronchial biopsies, and found all but one had NSE-like immunoreactivity. No such immunoreactivity was found in 26 serous fluids and 19 biopsies from non-small-cell carcinomas. It is suggested, therefore, that immunostaining for NSE is a valuable aid to the cytological diagnosis of small cell carcinoma of the lung.

Ascitic Fluid↗
PubMed1984

The potential of the immunogold-silver staining method for paraffin sections.

The immunogold-silver staining technique is shown to be of great value in the detection of regulatory peptide-containing nerves and endocrine cells in routinely fixed, paraffin-wax-embedded tissues. The method appears to be better for this system than peroxidase anti-peroxidase (PAP) which can yield poor or variable results. Antibodies to regulatory peptides, including calcitonin gene-related peptide (CGRP), substance P, neuropeptide tyrosine (NPY), glucagon, pancratic polypeptide, and somatostatin 14 and 28, as well as to neurofilaments, neuron-specific enolase (NSE) and S-100, were used on sections of a variety of tissues from rat and pig including respiratory tract, skin, gut, pancreas, vagina, uterus, fallopian tube and kidney. In all cases, stronger immunostaining of nerves was obtained with the immunogold-silver technique than with PAP. The inherent density of the staining was also found to improve the visibility of endocrine cells in the section, and to permit the use of routine histological stains for counterstaining. As immunogold-silver staining is sensitive, rapid, cheap and avoids hazardous reagents, we feel it has great potential for the immunostaining of nerves and endocrine cells that contain regulatory peptides in routinely fixed and embedded tissues and may prove useful in pathology.

Animals↗
PubMed1983

Distribution of bombesin, somatostatin, substance-P and vasoactive intestinal polypeptide in feline and porcine skin.

The content and distribution of several regulatory peptides in the skin of cats and pigs, freshly obtained at surgery, have been investigated. Immunoreactive bombesin was evenly distributed at low concentrations in both species, being below the detection limit in the body and nose of the cat, and showing a peak value of 1.6 +/- 0.7 pmol/g in the tip of the pig's ear. Similar concentrations of somatostatin-immunoreactivity (-IR) were found but greater regional variation occurred in the pig with a low in the mid back of 0.4 +/- 0.1 and the highest value in the snout of 3.1 +/- 0.8. Substance-P-IR in the pig showed a marked variation in concentration, apparently parallelling skin sensitivity, with a low in the back of 0.4 +/- 0.7 and higher values around the anus (8.1 +/- 1.6), legs (6.8 +/- 1.8) and snout (13.5 +/- 3.6) whilst in the cat values ranged from 0.3 +/- 0.06 in the body to 5.0 +/- 0.9 in the front footpads. In contrast, vasoactive intestinal polypeptide (VIP)-IR showed greater variability in the cat, being below the assay's detection in the body and highest in the front and rear footpads (17 +/- 7 and 29 +/- 6 respectively), but in the pig most regions exhibited low concentrations with the exception of the snout which peaked at 12.0 +/- 5.0. Immunocytochemical localisation showed the peptides to be present in nerve fibres. Substance-P-IR was particularly localised in the snout of the pig just below the epithelium while VIP-IR was more concentrated in deeper layers, often associated with sweat glands and blood vessels.

Animals↗
PubMed1983

The value of S-100 immunostaining as a diagnostic tool in human malignant melanomas. A comparative study using S-100 and neuron-specific enolase antibodies.

The brain proteins S-100 and neuron-specific enolase have been reported by separate groups to be present in human malignant melanomas. There is no systematic study comparing the occurrence of these proteins in the same tumour specimens. We have examined 33 primary malignant melanomas, including 5 which were amelanotic, and 25 metastatic melanomas using immunohistochemical methods with specific, non-cross-reacting antibodies to S-100 and NSE. We found S-100 immunoreactivity to be present in all cases but one, whereas NSE immunoreaction was very weak and patchy, and present in only 6 cases. S-100 immunoreactivity was not demonstrated in 40 control tumours, either primary or metastatic in skin, including basal- and squamous-cell carcinomas, spindle-cell sarcomas, lymphomas and Merkel cell tumours. All intradermal (n = 4) and compound (n = 1) naevi were positive for S-100, 2 blue naevi showing much less reaction. NSE immunoreactivity was detected in Merkel cell tumours (n = 8), undifferentiated (n = 2) and small cell (n = 1) carcinomas, and all melanocytic naevi. It is suggested therefore that antibody to S-100 is the reagent of choice for demonstration of melanocytic tumours, and may be especially valuable in the diagnosis of amelanotic melanoma or metastatic tumours of doubtful origin where melanoma is suspected.

Fluorescent Antibody Technique↗
PubMedDate not supplied

Deficiency of calcitonin gene-related peptide in Raynaud's phenomenon.

Skin biopsy samples from the fingers of nine patients with primary Raynaud's phenomenon, nine with the disorder associated with systemic sclerosis, and eleven healthy controls were examined by immunocytochemistry. There were no differences between the groups in the distribution of PGP 9.5 (a pan-neuronal marker) immunoreactivity, but there was a significant reduction in the number of calcitonin gene-related peptide (CGRP) immunoreactive neurons in the skin of patients with primary Raynaud's phenomenon and those with systemic sclerosis. These findings implicate dysfunction of the CGRP neurovascular axis in the pathophysiology of Raynaud's phenomenon.

Adult↗
PubMedDate not supplied

Induction of nitric oxide synthase in asthma.

Nitric oxide (NO) is a mediator of vasodilatation and bronchodilatation synthesised from L-arginine by the enzyme NO synthase, which is either constitutive or induced by lipopolysaccharides and/or cytokines. The presence and function of NO synthase in normal or diseased lung is not yet clear. Asthma is characterised by bronchial hyperresponsiveness, epithelial damage, inflammation, and increased cytokine production. To investigate the presence of NO synthase in asthma, we immunostained bronchial biopsies from non-steroid-treated people with asthma and non-asthmatic controls with specific polyvalent antisera to purified inducible NO synthase and to a selected peptide sequence of the same enzyme. Immunoreactivity was seen in the epithelium and some inflammatory cells in 22 of 23 biopsies from people with asthma, but in only 2 of 20 controls. To assess the relation of cytokines to NO synthase induction, bronchial epithelial cells in culture were stimulated with tumour necrosis factor (TNF alpha). Inducible enzyme immunoreactivity was found only in the treated cells. The existence of inducible NO synthase in human lungs suggests that increased production of NO, probably induced by cytokines, may be relevant to the pathology of asthma.

Adult↗
PubMedDate not supplied

Epithelially derived endothelin and nitric oxide in asthma.

Immunohistochemical staining of endobronchial biopsies has identified increased expression of the 21-amino-acid peptide endothelin (ET) and the inducible form of the enzyme nitric oxide synthase (iNOS) within the airway epithelium in asthma. Elevated concentrations of ET are also recovered in bronchoalveolar lavage fluid from asthma patients. iNOS generates the gas nitric oxide from L-arginine, and elevated levels of NO in exhaled air have been described in asthma. ET is a potent bronchoconstrictor and levels of ET in lavage and resting airflow obstruction are correlated. The effects of ET on bronchomotor tone may be modified by NO as this is a bronchodilator. The relative balance between ET and NOS may thus contribute to resting bronchomotor tone. ET also stimulates fibroblast proliferation, collagen gene expression and through its inhibitory actions on collagenase will promote airway wall collagen deposition and contributes to airway wall thickening which underlies bronchial hyperresponsiveness. The regulation of these epithelial events may thus be important to the control of asthma.

Amino Acid Oxidoreductases↗