Only few workers exposed to wood dust are detected with specific IgE against pine wood.

BACKGROUND: Our aim was to investigate the frequency of pine allergy in woodworkers with respiratory symptoms and to identify high molecular weight allergens in pine wood extracts. METHODS: In a cross-sectional study we examined work-related respiratory symptoms in 2033 furniture workers and 474 controls by questionnaires. Clinical examination was performed in 365 wood dust exposed and 116 nonexposed subjects. Blood samples were collected for measuring pine-specific immunoglobulin (Ig)E by an immunoassay and Western blots. RESULTS: Eleven exposed and three nonexposed subjects had pine-specific IgE. In the group with clinically defined asthma eight persons (5.4%) had pine-specific IgE compared with six persons (1.8%) in the group without asthma (P < 0.05). In the groups with and without respiratory symptoms, 13 (3.8%) and one (0.7%) subject, respectively, had pine-specific IgE (P = 0.06). Western blots demonstrated pine-specific IgE to components in the molecular range of 14 - 100 kD in eight samples (all wood dust exposed). Five samples had pine-specific IgE against components in a 43 - 59 kD zone and against two bands at 27 and 29 kD that are candidates for major allergens. CONCLUSION: Some workers in the Danish furniture industry are specific IgE sensitized against pine wood dust. Pine-specific IgE probably explains a minor part of the respiratory symptoms in workers exposed to pine wood dust.

Soluble intercellular adhesion molecule and C-reactive protein as early markers of infection in newborns.

In order to find a reliable early marker of infection in newborns a study with simultaneous determination of soluble Intercellular Adhesion Molecule-1 (sICAM-1) and C-Reactive Protein (CRP) was planned. Prospectively 90 babies < 5 days of age suspect of infection were included. Retrospectively this population was classified into an "infected" group (n = 45) and a "non-infected" group (n = 45). For each of these two groups we calculated the sensitivity, specificity and predictive values of sICAM-1 and CRP as early markers of infection. We determined the best cut-off level for sICAM-1 to be 300 micrograms/l and for CRP 5 mg/l. As a biochemical test for infection in the newborns the sensitivity and negative predictive value for CRP were 0.69 and 0.73 respectively. When sICAM-1 was added and CRP and s-ICAM-1 were used in combination the sensitivity improved significantly to 0.93, p < 0.01 and the negative predictive value improved to 0.92, p < 0.05. In normal 5-8 days old babies' sICAM-1 was significantly higher than at birth (cord blood), p < 0.0001. In conclusion, sICAM-1 and CRP in combination are better than CRP as a primary test for identification of infection in babies < 5 days of age.

False negative anti-HIV-1/HIV-2 ELISAs in acute HIV-2 infection.

Since HIV-2 was isolated in 1986, only 1 case of acute HIV-2 infection has been reported. We have identified another patient with primary HIV-2 infection. Follow-up samples were requested from the patient due to discrepant results. The HIV-2 infection was confirmed with HIV-2-specific proviral DNA amplification by PCR. The HIV-2 seroconversion panel obtained was used to evaluate the sensitivity of both combined and specific ELISAs currently in use in Europe, and to investigate the Western-blot patterns on both HIV-1-and HIV-2-specific Western blots. The window period was determined to be less than 37 days with the most sensitive assays. A remarkable difference in sensitivity to HIV-2 antibodies in acute HIV-2 infection was found in combined HIV-1/HIV-2 ELISAs. Three out of the 4 combined sandwich ELISAs appeared to be less sensitive than the indirect ELISAs in HIV-2 seroconversion, leading to a prolonged window period. One HIV-2-specific ELISA was also negative on the first sample, but positive on the second sample. In the HIV-2 Western blot, early reaction with HIV-2-specific env and gag proteins was seen, whereas the HIV-1 Western blot on the first sample revealed gag (p24, p55) reactivity only.

Antinuclear antibodies and anti-DNA antibodies in scleroderma. A possible relationship between joint manifestations and increased antibodies in localized scleroderma.

Antinuclear antibodies (ANA), including anti-DNA antibodies, and rheumatoid factors (RAT, Waaler-Rose) were determined prospectively during a 3-year period in 40 patients with localized scleroderma (LS) compared with 77 patients with generalized scleroderma (GS). ANA were increased in 26% of patients with LS, and in 47% with GS, anti-DNA antibodies in 23% of patients with LS, and in 34% with GS. Thus, the anti-DNA antibody level was lower compared with the known level in systemic lupus erythematosus. Rheumatoid factors were present in 6-7% of patients with LS, and in 14-15% of patients with GS. Increased antinuclear antibodies were not associated with any specific type of localized scleroderma, nor with internal disorders, and no case of clinical overlap to discoid or systemic lupus erythematosus was observed. However, six patients with localized scleroderma and complaints of arthralgia all presented increased antibodies, and one patient showed overlap to rheumatoid arthritis. It is suggested that increased ANA and anti-DNA antibodies in localized scleroderma, associated with joint manifestations, represents a systemic component in this type of scleroderma, with activation of the immune system and similarities with generalized collagen diseases.

Circulating endogenous vasoactive intestinal polypeptide (VIP) in patients with uraemia and liver cirrhosis.

The concentration of vasoactive intestinal polypeptide (VIP) in peripheral venous plasma was median 6.0 pmol l-1 (range 0-20) in 112 normal subjects. In fifty-three patients with decreased kidney function plasma VIP was significantly increased (median 15.0 pmol l-1, range 0.5-70, P less than 0.0001) and positively correlated to serum creatinine concentration (r = 0.51, P less than 0.001). In 133 patients with liver cirrhosis peripheral venous VIP was slightly elevated (median 7.0 pmol l-1 range 0-86, P less than 0.01). Samples obtained during a central venous catheterization showed significant renal extraction of circulating VIP in control subjects (median extraction fraction 23%, P less than 0.05, n = 6) and in patients with cirrhosis (median 60%, P less than 0.02, n = 8), but not in uraemic patients (median 0%, NS n = 5). In control subjects and patients with cirrhosis the concentration of VIP in the hepatic vein was significantly below that of systemic plasma (-42%, P less than 0.05, n = 6 and -45%, P less than 0.01, n = 10, respectively). On the contrary, in uraemic patients hepatic venous VIP was almost similar to systemic VIP (-4%, NS, n = 7). The results indicate that in normal subjects and patients with cirrhosis both the liver and kidneys are involved in the biodegradation of VIP. The elevated level of circulating VIP in uraemic patients may in part be due to decreased renal and hepatic biodegradation but increased neuronal release of VIP, especially in the splanchnic system, may also contribute to the increased plasma VIP in this condition.

VIP binding sites on synaptosomes from rat cerebral cortex: structure-binding relationship.

The structural requirements for VIP interaction with receptors on synaptosomes from rat cerebral cortex was investigated by the ability of VIP and VIP fragments, secretin analogues and fragments, peptides of the VIP/secretin family and several other regulatory peptides to inhibit specific 125I-VIP binding. Only large VIP fragments interacted with the VIP receptors with potencies relative to VIP ranging from 0.9-0.006%. The rank order of inhibition was: VIP 7-27 greater than VIP 11-28 greater than VIP 1-22-NH2 greater than VIP 16-28. Shorter fragments: VIP 18-28; VIP 18-28-NH2; VIP 19-28; VIP 21-28; VIP 22-28; VIP 1-18; VIP 1-18-NH2; VIP 1-10-NH2; VIP 1-6; VIP 16-20 and VIP 16-19 had no effect. Secretin fragments and analogues inhibited 125I-VIP binding with potencies of 2.2-0.01% relative to VIP in the order; secretin greater than (Ala4, Val5) secretin greater than (D-Ala4) secretin greater than (D-Phe6) secretin greater than secretin 5-27 greater than secretin 14-27. Other peptides of the VIP/secretin family inhibited 125I-VIP binding with potencies of 200-1%; avian VIP greater than porcine VIP greater than PHI = secretin greater than human GRF, whereas glucagon and GIP showed no inhibition. Among twenty-five other regulatory peptides only avian PP and somatostatin were inhibitors with relative potencies of 0.02% and 0.03%, respectively. In conclusion it may be emphasized that the intact VIP molecule is essential for VIP interaction with its receptors in the rat brain cortex.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of pregnancy and sex steroids on concentration, motor effect and receptor binding of VIP in the rabbit female genital tract.

The influence of sex steroid and pregnancy on the tissue concentration, uterine motor effect and receptor binding of VIP has been studied in the female genital tract of pregnant rabbits and oophorectomized rabbits during progesterone and/or oestrogen substitution. The concentration of immunoreactive VIP was high in the vagina and cervix, and lower in the uterine body of both pregnant and non-pregnant rabbits. A significant decrease in the VIP concentration (pmol/g wet weight) of the uterine body was observed toward term of pregnancy. The total uterine content of VIP, however, seems unchanged. Treatment of oophorectomized rabbits with ovarian steroids had no effect on the VIP concentration. The sensitivity for and potency of VIP on the relaxation of uterine muscle was significantly higher in oophorectomized rabbits treated with a combination of progesterone and oestrogen than in control rabbits. No difference was observed between non-pregnant and pregnant rabbits. The degradation and binding affinity for 125I-labelled VIP was highest in oophorectomized rabbits substituted with both oestrogen and progesterone. In the pregnant rabbits, the amount of receptors was decreased near term. In conclusion, sex steroids are able to influence the motor effect of VIP at the receptor level, but have no effect on the VIP concentration in the female genital tract.

The regional distribution of receptors for vasoactive intestinal polypeptide (VIP) in the rat central nervous system.

The regional distribution of receptors for vasoactive intestinal polypeptide (VIP) was studied in the rat central nervous system (CNS). The specific binding was highest in cerebral cortex, limbic forebrain and cerebellum, whereas moderate to low binding was found in hypothalamus, thalamus, brainstem and pituitary. The lowest binding was observed in pons and spinal cord. Scatchard analysis showed curvilinear plots with upward concavity, which was interpreted as two classes of binding sites. The Kd values were similar in all regions and calculated as 2.4 and 62 nmol/liter, respectively. The variations of specific [125I]VIP binding were due to differences in the total amount of receptors and were in the range of 1.7-8.6 pmol per mg protein. The regional distribution of VIP receptors was parallel with the occurrence of VIP-containing nerve terminals with exceptions of cerebellum, olfactory areas and nucleus caudatus, where a greater number of receptors than expected from the VIP content was found. In these regions, VIP may interact with receptors for a different, but homologous neuropeptide. In conclusion, the regional distribution of VIP receptors in CNS gives further evidence for the role of VIP as a central neurotransmitter.

Development of binding sites for vasoactive intestinal polypeptide in mouse cerebral cortex and cultured cortical neurons.

Binding sites for vasoactive intestinal polypeptide (VIP) and the content of immunoreactive VIP were measured in the foetal and neonatal mouse brain cortex and primary cultures of foetal murine brain-cortical neurons and astrocytes. The amount of cortical VIP binding sites and the concentration of immunoreactive VIP were low before birth, but increased postnatally reaching adult level at about 3 weeks of age. In cultures, a similar rise in neuronal binding sites occurred after 10 days, whereas the VIP concentration remained lower than in adult brain cortex. No binding sites or immunoreactive VIP could be detected in cultured astrocytes. The VIP binding sites were heterogeneous both in developing brain cortex and cultured neurons, and consisted of two classes of binding sites. The high affinity constants (2.5-2.9 nM) as well as low affinity constants (50-76 nM) were unchanged during the development, whereas the number of binding sites increased. In cultured neurons the binding constants were similar to those found in the adult mouse brain cortex. The peptide specificity for the VIP binding was similar in brain cortex and neuronal culture. In conclusion, the maturation of murine brain cortical neurons is accompanied by the development of synaptosomal VIP receptors in support of the neurotransmitter function of VIP.

Insulin receptors in rat brain cortex. Kinetic evidence for a receptor subtype in the central nervous system.

Binding kinetics of porcine 125I-insulin were studied in synaptosomal and microsomal fractions of rat brain cortex. Receptor binding was temperature- and pH-dependent with optimum at 4 degrees C and pH 8.0-8.3. At 15 degrees C, steady state binding was heterogenous, and Scatchard analysis revealed two classes of receptors with Kd of 2 nmol/l and 40 nmol/l in amounts of 50 pmol/g and 200 pmol/g of membrane protein. Dissociation kinetics were biexponential with T1/2 of about 5 min and 180 min, and in contrast to other cell-types, not influenced by negative cooperativity. No receptor-mediated insulin degradation was detectable at 37 degrees C in the presence of bacitracin. Insulin analogues inhibited 125I-insulin binding with potencies relative to porcine insulin (%): human insulin 100, rat insulin (I + II) 71, coypu insulin 47, rat multiplication stimulating activity 8, porcine proinsulin 5, among which the three last values were significantly higher than in rat liver and fat cells. No competition was observed with porcine relaxin and mouse nerve growth factor up to about 1 mumol/l. Receptors were present in all regions of central nervous system with highest concentrations in the cerebral cortex, cerebellum and olfactory bulb, and lowest in the pons, medulla oblongata and spinal cord. In conclusion, insulin receptors in rat brain cortex are functionally different from other tissues regarding the insulin specificity and the absence of negative cooperativity. It is suggested that an insulin receptor subtype in rat brain mediates the growth activity of insulin on nerve cells.

VIP receptors in pig liver cells: binding characteristics and role in degradation.

The physiological role of VIP in the liver is controversial. VIP receptors are present, but their function in the metabolic regulation is uncertain. The interaction of porcine VIP with isolated cells from pig liver was studied with respect to receptor-binding, degradation and glycogenolytic action. In this model, VIP and liver showed homology of animal species. 1. Receptor-binding was heterogenous with Kd values of 10(-9) mol/l and 4 X 10(-8) mol/l, and a total amount of binding sites of 7 X 10(-11) mol per 10(9) cells. The peptide specificity showed that porcine and chicken VIP were equally potent in inhibiting receptor-bound 125I-VIP; secretin was about 30 times less potent; glucagon and somatostatin were ineffective. 2. Receptor-bound 125I-VIP was degraded since about 70% was released as radioactivity not reacting with VIP-antiserum. 3. Glucose-release was not stimulated by VIP (10(-6) mol/l) whereas the rate was increased two-fold by glucagon (10(-6) mol/l). In conclusion, VIP receptors in pig liver cells are different from other tissues regarding peptide specificity. It is suggested that receptor-binding mediates degradation of VIP by pig liver rather than metabolic effects.

Iron, copper, zinc and selenium in human liver tissue measured by X-ray fluorescence spectrometry.

The content of iron, copper, zinc and selenium was measured by energy-dispersive X-ray fluorescence (XRF) spectrometry in normal liver tissue obtained at autopsy from 16 females and 12 males 46-87 years old. The precision of the XRF analysis, expressed by the coefficient of variation was: iron, 1.8%; copper, 3.2%; zinc, 1.0%; and selenium, 26.7%. In large liver samples, mean amount-of-substance contents of elements in dry liver tissue were: iron, 16.95 mmol/kg (range 7.90-27.31 mmol/kg); copper, 0.33 mmol/kg (0.08-0.76 mmol/kg); zinc, 5.12 mmol/kg (2.92-9.47 mmol/kg); selenium 0.02 mmol/kg (less than 0.004-0.04 mmol/kg). Furthermore the amounts of iron, copper and zinc were measured in wet-ashed Menghini needle biopsy specimens taken from the centre of 20 large liver samples. There was good agreement between results obtained in biopsy specimens and large samples concerning iron (r = 0.96, P less than 0.001) and zinc (r = 0.97, P less than 0.001), but not concerning copper (r = 0.66, P less than 0.01). XRF analysis appears to be a convenient method for element analysis in liver tissue and for measurement of iron and zinc in needle biopsy specimens.

Escherichia coli antibodies in alcoholic liver disease. Correlation to alcohol consumption, alcoholic hepatitis, and serum IgA.

In 41 patients with alcoholic liver disease, antibodies to 12 common Escherichia coli O antigens (expressed as number of O antibody reactions with an agglutination titre of greater than or equal to 40) and to immunoglobulins IgG, IgA, and IgM were studied for 8 weeks. In 18 patients (8 with cirrhosis, 10 with fatty liver) who continued drinking during this period no significant changes were found. In 23 patients (11 with cirrhosis, 12 with fatty liver) who stopped or reduced drinking, a significant decrease in the levels of E. coli O antibodies and IgA was found (p less than 0.05 and p less than 0.01, respectively). In these 41 patients and in an additional 43 patients with alcoholic liver disease the amount of E. coli O antibodies was compared with type of histological lesion. The levels of E. coli O antibodies were significantly higher in cirrhosis with alcoholic hepatitis (22 cases) than in cirrhosis without alcoholic hepatitis (17 cases) (p less than 0.05). In these 17 patients antibody levels were significantly higher than in 41 patients with fatty liver without alcoholic hepatitis (p less than 0.02). In all patients a significant correlation between the number of positive reactions to E. coli O antigens and serum IgA concentration was found (p less than 0.01). No microbes were cultured from the liver biopsies, and no E. coli O antigens were demonstrated in the liver tissue by immunohistochemistry. Our results support the hypothesis that the high levels of E. coli O antibodies in alcoholic liver diseases are due to failure of the liver to extract circulating antigens and gut-derived endotoxins.(ABSTRACT TRUNCATED AT 250 WORDS)

Multiplicity of receptors for vasoactive intestinal polypeptide (VIP): differential effects of apamin on binding in brain, uterus and liver.

Apamin is a neurotoxic octadecapeptide from bee venom, which has been shown to inhibit the non-adrenergic, non-cholinergic inhibitory innervation of the smooth muscle of the gut. Since vasoactive intestinal polypeptide (VIP) has been proposed as a possible inhibitory neurotransmitter, the effect of apamin on the receptor binding of 125I-VIP was studied using the following assays: (1) isolated synaptosomes from rat cerebral cortex, (2) crude plasma membranes from hog uterine smooth muscle, and (3) purified plasma membranes and isolated hepatocytes from hog liver. Apamin inhibited the receptor-bound 125I-VIP on membranes from brain or myometrium, although the binding affinity was 100-1000 times lower than for VIP. The displacement curves for VIP and apamin were parallel suggesting that apamin interacts with both the low and high affinity VIP receptors. In membranes and cells from liver, apamin was unable to displace receptor-bound 125I-VIP in concentrations up to 50 mumol/l. The findings suggest that the VIP receptors in liver are different from those in the brain cortex and myometrium.

Immunoreactive trypsin and pancreatic isoamylase activity in serum of patients with chronic renal failure or hepatic cirrhosis.

In 121 patients with either liver cirrhosis or chronic renal failure, abnormal values for the concentrations of two pancreatic enzymes in serum were a frequent finding. In renal insufficiency a decreased rate of enzyme elimination is the most likely cause of the above-normal values we observed for serum immunoreactive trypsin and pancreatic isoamylase activity. As for patients with liver cirrhosis, we believe that changes in entrance rates into the blood--i.e., an affected pancreas--is a likely explanation of the abnormally high values we often found for these serum enzymes.

Vasoactive intestinal polypeptide (VIP) in cirrhosis: arteriovenous extraction in different vascular beds.

The concentration of vasoactive intestinal polypeptide (VIP) was determined in peripheral venous plasma from 136 patients with liver cirrhosis without gastrointestinal bleeding or coma and from 112 controls. In eight patients (cirrhosis, six; fibrosis, one; steatosis, one) arteriovenous extraction or release of VIP was measured during catheterization at four locations: brain, lower limb, intestine-liver, and kidney. The mean concentration of VIP in peripheral venous plasma from patients with cirrhosis was 9.4 pmol/l (median, 7.0; range, 0-86), which was significantly higher than that of the controls, who had a mean of 6.2 pmol/l (median, 6.0; range, 0-20, P less than 0.01). No significant extraction or release of VIP could be detected across the vascular bed in brain or lower limb. A significant arterio-hepatovenous VIP extraction ratio (mean, 0.43; range, 0.05-0.87) confirmed at net splanchnic elimination of VIP from extra-splanchnic areas and from porto-systemic shunting of VIP in cirrhosis. The net splanchnic elimination rate of VIP was estimated to be about 3 pmol/min. The concentration of VIP in ascitic fluid was on the average three times that of arterial plasma. In conclusion, VIP is significantly elevated in peripheral plasma from patients with cirrhosis, probably due to porto-systemic shunting and/or compromised hepatic elimination. Hepatic elimination is still likely to account for the inactivation of most of the VIP escaping from the neurosynapses throughout the body in patients with cirrhosis without coma.

Serum ethanol estimations in the control of alcohol abstinence in patients with liver disease.

Eighty-eight patients with a non-alcoholic and 105 patients with an alcoholic liver disease were warned against alcohol consumption. On three consecutive ambulatory visits, serum ethanol was measured and compared with patients' admission of alcohol intake. None in the non-alcoholic group had a positive serum ethanol test, whereas 60 samples from 40 patients with alcoholic liver disease were positive. The serum ethanol values were higher in women than in men. Continuation of drinking was unrelated to sex, age, or type of alcoholic liver disease. Twenty-seven of the 40 patients with ethanol in serum denied alcohol consumption. The reliability of the patients was unrelated to sex, age, or type of alcoholic liver disease. Serum ethanol was more valuable than aspartate aminotransferase, alkaline phosphatase, bilirubin, and coagulation factors in pointing out the patients who continued drinking.