Ultrastructural evidence of brain mast cell activation without degranulation in monkey experimental allergic encephalomyelitis.

Experimental allergic encephalomyelitis (EAE) is an animal model for the human demyelinating disease multiple sclerosis (MS). Increased permeability of the blood-brain barrier (BBB) precedes the development of clinical or pathologic findings in MS and may be induced by perivascular brain mast cells secreting vasoactive and proinflammatory molecules. Brain mast cells were investigated ultrastructurally in acute EAE of the non-human primate common marmoset Callithrix jacchus, which develops a mild neurologic relapsing-remitting course. Control diencephalic samples contained perivascular mast cells with mostly intact electron dense granules. In contrast, EAE samples had marked demyelination and mast cells with numerous altered secretory granules; their electron dense content varied in amount and texture with a "honeycomb" or "target" appearance, but without degranulation. These changes were evident even before the development of any clinical symptoms and suggest that brain mast cells may be involved in EAE, and possibly MS, through a unique process that may involve selective secretion of molecules able to disrupt the BBB.

[Cerebral adrenomyeloneuropathy as a late type of adrenoleukodystrophy: case report]. / Mózgowa odmiana adrenomyeloneuropatii jako pózna postac adrenoleukodystrofii. Opis przypadku.

Adrenoleukodystrophy (ALD) is a hereditary disease related to abnormalities in the structure and function of peroxisomes. The nature of disorder arises from defective process of beta-oxidation of very-long-chain fatty acids and their accumulation in various organs. ALD may present as a few phenotypes urologic symptomatology of which depends on proportions of the brain, spinal cord and peripheral nerves affection. Below, we present a case (and its family) of a cerebral type of adrenomyeloneuropathy--a rare form of adrenoleukodystrophy, and discuss its clinical manifestation as well as biochemical, hormonal, electrophysiological, neuropsychological and magnetic resonance imaging (MRI) diagnostics.

Morphological and functional demonstration of rat dura mater mast cell-neuron interactions in vitro and in vivo.

Mast cells derive from a distinct bone marrow precursor and mature in tissues under the influence of stem cell factor, nerve growth factor (NGF) and certain interleukins. Intracranial mast cells first appear in the meninges and are located perivascularly close to neurons. They can be activated by antidromic stimulation of the trigeminal nerve, as well as by acute immobilization stress. Substance P (SP) and corticotropin-releasing hormone (CRH) are particularly potent in stimulating mast cell release of vasoactive, inflammatory and nociceptive molecules. These findings have suggested that mast cells may be involved in neuroinflammatory conditions, such as migraines. In this study, dura mast cells were shown to have characteristics of connective tissue mast cells (CTMC) as they contained histamine, heparin and rat mast cell protease I (RMCP-I). Mast cells were localized close to SP-positive neurons immunocytochemically and mast cell-neuron contacts were also documented using scanning electron microscopy. Dura stimulated by SP and carbachol in situ released histamine. Preincubation of dura with estradiol slightly augmented histamine release by SP, an effect possibly mediated through estrogen receptors identified on dura mast cells. Acute stress by immobilization led to dura mast cell degranulation which was prevented by pretreatment with a neutralizing antibody to CRH or a CRH receptor antagonist. The present results further clarify the biology of intracranial mast cells and support their involvement in the pathophysiology of migraines which are precipitated or worsened by stress.

Differential effect of histamine 3 receptor-active agents on brain, but not peritoneal, mast cell activation.

The activation of presynaptic histamine 3 (H(3)) receptors inhibits the release of histamine and other neurotransmitters from central nervous system neurons. Rat brain mast cells (MCs) release histamine and 5-hydroxytryptamine (5-HT) in response to neuropeptides and neurotransmitters secreted from adjacent neurons. Dura MCs also degranulate in response to antidromic trigeminal nerve stimulation and with acute psychological stress. Such findings have implicated brain MCs in certain neuroinflammatory disorders, such as migraines. We investigated the ultrastructural appearance of control and stimulated thalamic/hypothalamic (brain) MCs before and after treatment with the H(3) receptor agonist N(alpha)-methylhistamine (N(alpha)-mH) and the H(3) receptor antagonist thioperamide (Th). Ultrastructural investigation of brain MCs stimulated with compound 48/80 revealed extensive intragranular changes that paralleled 5-HT secretion but without degranulation by exocytosis typical of connective tissue MCs. N(alpha)-mH significantly reduced these morphological changes, as well as 5-HT release from brain MCs and neurons stimulated with KCl; conversely, Th augmented both histamine and 5-HT release from brain neurons and MCs. Neither N(alpha)-mH nor Th had any effect on peritoneal MCs. Simultaneous addition of both drugs largely antagonized each other's effects on brain MC activation and 5-HT secretion. Ultrastructural observations and lack of lactic dehydrogenase release in the perfusate excluded any cytotoxic effect. The ability of H(3) agonists to inhibit brain MC activation, as well as secretion of 5-HT from both brain MCs and neurons, may be useful in the management of migraines.

Mast cell activation in children with migraine before and after training in self-regulation.

Migraine may affect as many as 9% of all schoolchildren and often presents with abdominal symptoms of pain, nausea, and vomiting. Even though the pathophysiology of migraine remains unknown, self-regulation techniques appear to be more effective in prevention of childhood migraine than conventional pharmacotherapy which is often associated with adverse effects. Mast cells have been implicated in the pathogenesis of migraine in adults, but have not been previously studied in children with migraine. Mast cells are found close to the vessels and nerves in the meninges where they can release multiple vasoactive, neurosensitizing, and pro-inflammatory mediators. Therefore, we investigated whether children with migraine may have increased urinary levels of mast cell mediators and whether practicing relaxation imagery exercises has an effect on the frequency of headache, as well as on mast cell activation. Urine was collected for 24 hours from children with and without migraine after a 5-day amine-restricted diet. Children with migraine also collected urine during migraine episodes. The mean levels of urinary histamine, its main metabolite, methylhistamine, and the mast cell enzyme, tryptase, were higher in children than generally found in adults, but they did not differ statistically in any of the categories studied. However, in 8 of 10 children who practiced relaxation imagery techniques and successfully reduced the number of migraines, the urine tryptase levels were also significantly lower. There was no relationship between successful practice and sex or age of the child. These results suggest that stress may activate mast cells which could be involved in the pathophysiology of migraine.

[The influence of multiple histamine inhalation on nonspecific airway hyperresponsiveness to histamine in patients with bronchial asthma]. / Wplyw wielokrotnego wziewania histaminy na nieswoista nadreaktywnosc oskrzeli na histamine u chorych na astme oskrzelowa.

The study was performed in 19 patients with stable mild/moderate asthma aged from 16 to 66. Nonspecific airway hyperresponsiveness (AH) to histamine was measured according to Cockcroft's et al. method and expresses as PC20H in mg/ml. The study was controlled by inhalation of PBS. The histamine or PBS challenges were performed three times at 45 min. intervals on the third day in case when the stability of AH on two preceding days was observed. The geometric means of PC20H (xg PC20H) during the consecutive three days did not change. They were 0.97, 0.92, 0.87 mg/ml (p > 0.05) respectively. Three times histamine challenges performed on the same day induced the decrease of AH (xg PC20H increased from 1.48 to 6.55 mg/ml) in 5 patients. In 4 other subjects the increase of AH was observed (xg PC20H decreased from 1.14 to 0.20 mg/ml). In 10 subjects the three times histamine challenges performed on the same day had no influence on AH to histamine.

[An increase of nonspecific airway hyperreactivity after repeated challenge with subthreshold doses of allergen in asthma patients]. / Wzrost nieswoistej nadreaktywnosci oskrzeli po wielokrotnej prowokacji podprogowa dawka alergenu u chorych na atopowa astme oskrzelowa.

The study was performed in 11 patients suffering from stable mild and moderate atopic asthma. Bronchial allergen challenges were performed according to Chai et al method. Airway hyperreactivity-AH (PC20H in mg/ml) was determined in each study day before subthreshold allergen inhalation dose, meaning the dose which did not induce asthmatic responses. Then FEV1 was measured in 2, 5, 15, 30 i 60 min. after allergen provocation throughout 4 to 9 consecutive days. A significant increase of PC20H was observed in 8 patients (xg PC20H in whole group decreased from 0.93 to 0.35 mg/ml after allergen challenges). The authors conclude that allergen inhalation challenge without asthmatic bronchial responses can cause an increase AH.

[Complications after BCG vaccination in the urban section of Lodz in the years 1994-5]. / Powiklania po szczepieniach BCG w aglomeracji lódzkiej w latach 1994-95.

The purpose of this paper was to evaluate the incidence of complications after BCG vaccination in children from urban area of Lódz in 1994-1995 and to give their pathological and prognostic interpretation on the basis of immunological and Groër allergometric examinations. The obtained data demonstrate that postvaccinal complications occurred in 46 children, that is 0.7/1000 of vaccinated population. They were observed mainly in newborns (45.7%), whereas they were particularly rare in revaccinated six-year-old children (13%) and schoolchildren (8.7%). In half of the cases there was an evidence of ulceration and suppuration in the site of vaccination, in another half--suppuration of local lymph nodes with or without fistula. Immunological and allergometric examinations were carried out in 21 children with post-vaccinal complications and 21 children with normal post-vaccinal period. In both groups the following were the subjects of evaluation: values of B and T lymphocytes and their CD4 and CD8 subpopulations, lymphocytes proliferative response to mitogenic PHA doses and tuberculin, as well as IgG, IgA and IgM levels. Immunological and allergometric examinations indicated that immunosuppression was neither the cause nor the effect of BCG complications.

[The effect of nedocromil sodium on spontaneous production of histamine releasing factor (HRF) by mononuclear blood cells in patients with nonatopic bronchial asthma]. / Wplyw nedokromilu sodu na samoistne wytwarzanie, przez komórki jednojadrowe krwi obwodowej, czynnika uwalniajacego histamine z bazofili (HRF) u chorych na nieatopowa astme oskrzelowa.

The aim of the study was to evaluate the effect of 28-days antiinflammatory treatment with nedocromil sodium (Ned.sod.) (Tilade-8 mg/24 hours) on nonspecific bronchial hyperreactivity (PC20H in mg/ml) and spontaneous HRF activity production by mononuclear blood cells in patients with nonatopic bronchial asthma treated with beclomethasone dipropionate. The correlation between PC20H and HRF was also examined. The study was performed in 10 subjects with mild and moderate chronic, stable asthma in a double-blind, placebo-controlled way. Placebo and Ned. sod were given through 4 weeks in a randomized way with 8 weeks wash-out period. It was shown that Ned. sod. did not influence clinical asthma symptom scores, ventilatory parameters and PC20H. No changes in the spontaneous production of HRF activity were observed. The correlation between HRF activity and PC20H assessed 4 times was not significant. The authors conclude that studies with Ned. sod. in nonatopic asthma should be continued, but the dosage of the drug ought to be bigger and the time of treatment should to be longer.

Definitive characterization of rat hypothalamic mast cells.

Mast cells have previously been identified in mammalian brain by histochemistry and histamine fluorescence, particularly in the rat thalamus and hypothalamus. However, the nature of brain mast cells has continued to be questioned, especially because the electron microscopic appearance often shows secretory granule morphology distinct from that of typical connective tissue mast cells. Here we report that mast cells in the rat hypothalamus, identified based on metachromatic staining with Toluidine Blue, fluoresced after staining with berberine sulfate, indicating the presence of heparin. These cells were also positive immunohistochemically for histamine, as well as for rat mast cell protease I, an enzyme characteristically present in rat connective tissue mast cells. In addition, these same cells showed a very strong signal with in situ hybridization for immunoglobulin E binding protein messenger RNA. However, use of antibodies directed towards immunoglobulin E or its binding protein did not label any cells, which may mean either the binding protein is below the level of detection of the techniques used or that it is not expressed except in pathological conditions when the blood-brain barrier becomes permeable. At the ultrastructural level, perivascular mast cells contained numerous, intact, electron-dense granules which were labeled by gold-labeled anti-rat mast cell protease I. These results clearly demonstrate the presence of perivascular mast cells in the rat hypothalamus, where they may participate in homeostatic processes.

Changes of serum concentration of lipid peroxidation products in patients with pneumonia.

During lower respiratory tract infection, massive influx and activation of phagocytes is observed. Reactive oxygen species (ROS) released by macrophages and polymorphonuclear neutrophils (PMNs) kill microorganisms and cause damage to host tissues. One feature of this damage may be enhanced lipid peroxidation. Therefore, the aim of this study was to estimate the serum concentration of lipid peroxidation products in combination with clinical and biochemical indicators of inflammation in 32 patients with pneumonia. Serum concentration of lipid peroxides (CLP) and malondialdehyde (CMDA) was measured at Day 1, 4, 10 and 14 of observation, whilst chest radiography and routine blood analysis were performed at Day 1 and 14 during a 2 week treatment of lower airway infection. The CLP decreased during treatment (p < 0.05) from 0.059 +/- 0.024 to 0.043 +/- 0.017 (A532 nm) and the CMDA (p < 0.05) from 3.5 +/- 1.4 to 2.8 +/- 1.3 nmol.mL-1. A negative correlation between CLP and radiological regression (r = 0.49) and a drop in white blood cell count (WBC) (r = 0.39) was observed during the treatment. A positive correlation between CMDA and serum trypsin inhibitory capacity (STIC) (r = 0.47) and erythrocyte sedimentation rate (ESR) (r = 0.43) was found. Our data indicate that an enhanced process of lipid peroxidation occurs during pneumonia and that serum concentration of lipid peroxides returns to normal values quicker than the concentration of malondialdehyde during recovery. The use of antioxidants is suggested as an adjuvant treatment in patients with pneumonia.

[Chemotactic serum activity for neutrophils and eosinophils and spontaneous production of histamine releasing factor in patients with seasonal bronchial asthma treated with nedocromil sodium]. / Aktywnosc chemotaktyczna surowicy dla neutrofili i eozynofili oraz samoistna produkcja przez komórki jednojadrowe krwi obwodowej czynnika uwalniajacego histamine u chorych na sezonowa astme oskrzelowa leczonych nedokromilem sodu.

Nedocromil sodium (NS- Tilade) is an effective therapeutic agent against asthma and has been shown to exhibit antiinflammatory activity in vitro. The aim of this study was to assess the effect of NS on the serum neutrophil (NCA) and eosinophil (ECA) chemotactic activity and mononuclear cells-derived histamine releasing factor (HRF) activity in 14 patients with seasonal allergic asthma. The chemotactic activities, HRF and bronchial reactivity to histamine (PC20H in mg/ml) were determined before the grass-pollen season and at the end of the 14-day placebo treatment and 21-day NS treatment during the pollen-season. NCA and ECA were assessed using of the Boyden chamber methods. Blood samples for mononuclear cells culture were collected during the trial. The supernatants were assayed for HRF activity with basophils from a single donor. The NCA index was unaltered during the trial. There was a significant (p < 0.05) increase in the ECA after the NS treatment period (ECA index--2.5) compared with out of season and after placebo treatment (ECA index, 1.88 and 1.82). HRF activity increased after placebo compared with out of season (HRF % activity--38.62 before season, 47.61 after placebo p < 0.05) but NS did not revealed the further effect on HRF activity (49.81 p > 0.05). During placebo the significant decrease in PC20H from 1.81 mg/ml to 0.54 mg/ml (p < 0.05) was observed. NS did not have the effect on nonspecific bronchial reactivity--PC20H--0.74 mg/ml (p > 0.05). The correlation between PC20H and spontaneous production of HRF activity was not found. The results indicate that NS has some effect on inflammatory process which takes place in seasonal asthmatic patients during the natural allergic exposure.

[Bronchial hyperresponsiveness in patients with seasonal allergic rhinitis and the influence of loratadine and beclomethasone on this hyperresponsiveness]. / Wystepowanie nieswoistej nadreaktywnosci oskrzeli u chorych na sezonowy alergiczny neizyt nosa oraz wplyw loratadyny i beklometazonu na te nadreaktywnosc.

60 patients aged 16 to 46 years with seasonal allergic rhinitis were selected for study. Daily symptom scores of seasonal allergic rhinitis and nonspecific bronchial hyperresponsiveness to histamine (PC20H in mg/ml) before, during the pollen season and after 3 weeks of treatment with loratadine, beclomethasone dipropionate were evaluated. The control group received oxymetazoline. Nonspecific bronchial hyperresponsiveness for 11 patients (18.3%) before the seasons was observed. At the beginning of the season frequency of nonspecific bronchial hyperresponsiveness increased to 26.3% and to 36.8% after 3-week treatment course. Bronchial hyperresponsiveness was not related to any of the way of treatment. The patients treated with beclomethasone dipropionate and oxymetazoline showed significant relief of nasal symptoms. The patients without bronchial hyperresponsiveness showed lower value of symptom scores.

[The effect of bronchial allergen provocation on spontaneous production of histamine releasing factor (HRF) by mononuclear cells in patients with bronchial asthma]. / Wplyw wziewnej prowokacji alergenem na samoistne wytwarzanie czynnika uwalniajacego histamine z bazofili (HRF) przez komórki jednojadrowe krwi obwodowej u chorych na astme oskrzelowa.

The study was performed in 8 patients suffering from mild asthma. Bronchial allergen challenges with Dermatophagoides pretonyssinus were performed according to Chai et al method. Nonspecific bronchial reactivity to histamine (PC20H in mg/ml) were determined before the allergen challenge and in 90 min. and 24 hrs. after allergen provocation. Blood samples for mononuclear cells culture were collected before the trial and after the allergen challenge in 15, 90 min. and 8 and 24 hrs. The supernatants were assayed for HRF activity with basophils from a single donor. The significant increase of PC20H was observed xgPC20H in mg/ml 0.43 before allergen challenge and 0.08 and 0.125 after allergen provocation. Blood samples did not reveal any significant changes in HRF % activity--mean and SD before challenge test 47.7 16.2 and 49.9 15.0 in 15 min, 53.3 19.0 in 90 min, 56.6 15.8 in 8 hr, 52.5 20.4 in 24 hr after allergen provocation. The correlation between postallergen, nonspecific bronchial reactivity and spontaneous production of HRF activity was not found. The authors discuss the role of HRF in pathogenesis of nonspecific bronchial reactivity of asthmatic patients.

Stress-induced intracranial mast cell degranulation: a corticotropin-releasing hormone-mediated effect.

Stress is known to precipitate or worsen a number of disorders, such as migraines, in which mast cells are suspected of being involved by releasing vasoactive, nociceptive, and proinflammatory mediators. However, no functional association has been demonstrated yet between a migraine trigger and brain mast cell activation. Nontraumatic immobilization (restrain) stress has been shown to stimulate the hypothalamic-pituitary-adrenal axis and to cause redistribution of immune cells. Here, restrain stress caused degranulation in 70% of rat dura mast cells within 30 min, as shown both by light and electron microscopy. These morphologic findings were accompanied by cerebrospinal fluid elevation of rat mast cell protease I, but not II, indicating secretion from connective tissue type mast cells. Mast cell activation due to stress was abolished in animals that had been treated neonatally with capsaicin, indicating that neuropeptides in sensory nerve endings are involved in this response. Complete inhibition was also achieved by pretreating the animals ip with polyclonal antiserum to CRH. Mast cells in the dura were localized close to nerve processes containing substance P, but no CRH-positive fibers were identified even though these were found close to mast cells in the median eminence. This is the first time that stress is shown to activate intracranial mast cells; apparently through the sequential action of CRH and sensory neuropeptides. These findings may have implications for the pathophysiology and possible therapy of neuroinflammatory disorders such as migraines, which are induced or exacerbated by stress.

The value of C1 esterase inhibitor in patients with aspirin-sensitive urticaria.

The aim of the study was to evaluate the concentration and activity of C1 esterase inhibitor (C1 INH) in patients with aspirin-sensitive urticaria. C1 INH deficiency is the basis of hereditary angioneurotic edema. The study was performed on 32 subjects with aspirin-sensitive urticaria. The value of C1 INH in examined patients was the same as in the control group. There seems to be no coexistence of aspirin-sensitive urticaria and C1 esterase inhibitor deficiency.

Elevated mast cell tryptase in cerebrospinal fluid of multiple sclerosis patients.

Multiple sclerosis (MS) lesions are associated with infiltration of T lymphocytes and macrophages that appear to mediate myelin destruction and gliosis (scarring). Mast cells are located perivascularly in the brain, are juxtaposed to neurons, and have been shown to secrete vasoactive and inflammatory mediators in response to neuropeptides and direct nerve stimulation. Mast cells have been previously identified in MS lesions, are activated by myelin basic protein, and can participate in the regulation of blood-brain barrier permeability, as well as in myelin destruction. Here, cerebrospinal fluid from MS patients and controls with other neurologic diseases was assayed for histamine, its major metabolite methylhistamine, and the specific mast cell marker tryptase. Histamine and methylhistamine were not elevated in MS. However, the mast cell specific proteolytic enzyme tryptase was significantly elevated in MS, suggesting that mast cell activation may be involved in the pathophysiology of this disease.

[Effect of tobacco smoke on serum antioxidant activity]. / Wplyw dymu tytoniowego na aktywnosc antyoksydacyjna surowicy.

Oxidants of cigarette smoke and those released from phagocytes in smoker's lungs may inactivate alpha-1-antiproteinase. Thus, the "elastase-antielastase" imbalance may lead to lung tissue destruction and emphysema. Insufficient antioxidant protection, postulated by some authors might be an additional factor contributing to this process. The presented data on the influence of acute in vitro exposure to cigarette smoke in humans did not show difference in serum antioxidant activity (AOA) before and after the exposure. Attempts were made to evaluate the effect of whole cigarette smoke, its gas-phase and water-soluble phase on serum AOA in vitro. The results show that gas-phase leads to the depletion of serum AOA, whereas water-soluble phase exerts protective effect. Thus, at least part of oxidants might be inactivated in the stream of inhaled smoke. In conclusion, we doubt that serum AOA is influenced as a result of smoking and that the depletion of serum AOA is a decisive factor in the development of pulmonary emphysema.

[Course of non-specific bronchial reactivity to histamine after bronchospasm induced by allergen challenge in patients with bronchial asthma]. / Przebieg reaktywnosci oskrzeli na histamine po skurczu oskrzeli wywolanym wziewnie podanym alergenem u chorych na astme oskrzelowa.

Bronchial hyperresponsiveness is the characteristic feature of bronchial asthma. Inhalation of allergen can cause dual asthmatic response--early and late reaction. Thirteen patients with mild and moderate asthma underwent bronchial allergen challenge. Non-specific bronchial reactivity was measured before, 90 minutes, on the 2nd, 7th, 14th day after provocation. Peak Expiratory Flow was measured every hour during 12 hours after challenge to search for late phase reaction. Increased bronchial reactivity was discovered as early as 90 minutes after challenge, it was still observed on the 2nd and 7th but not on the 14th day after allergen provocation. No relationship was found between appearance of late asthmatic reaction and increased bronchial reactivity.