Serum CRP in patients with gout and effects of benzbromarone.

OBJECTIVE: C-reactive protein (CRP) is associated with increased risk for myocardial infarction, atherosclerosis, and peripheral artery diseases, while increased serum uric acid level is suggested to be independently associated with an increased risk of cardiovascular mortality. Accordingly, to investigate whether hyperuricemia is associated with serum CRP, we compared serum CRP levels between healthy subjects and patients with gout. In addition, we also examined whether benzbromarone has effects on serum CRP levels in patients with gout and the expression of CRP messenger RNA of CRP in the hepatoma cell line HuH7. METHODS: In the first experiment, 40 healthy males and 43 male patients with gout were enrolled, then blood samples were drawn from each after an overnight fast. In the second experiment, 42 male patients with gout were given uric acid-lowering therapy with benzbromarone. Blood samples were drawn after an overnight fast before and 1 year after beginning benzbromarone treatment. In the third experiment, the effects of benzbromarone on IL1beta-induced CRP expression were determined in HuH7 cells. RESULTS: Log serum CRP levels were not significantly different between the patients with gout and healthy subjects, while log serum CRP levels were decreased by 11% after benzbromarone treatment, as compared to the values before treatment (p < 0.01). In addition, log serum adiponectin levels were elevated by 2% after treatment (p < 0.01). Furthermore, our in vitro findings demonstrated that benzbromarone down-regulated IL1beta-stimulated CRP gene expression. CONCLUSIONS: These results suggest that hyperuricemia may not contribute to an increase in serum CRP level, while benzbromarone may have a favorable effect on CRP.

Effects of bovine milk ingestion on urinary excretion of oxypurinol and uric acid.

OBJECTIVE: Although allopurinol is a xanthine oxidase inhibitor, its overall effect may be due to the action of oxypurinol, a metabolite of allopurinol and another xanthine oxidase inhibitor, since the biological half-life of oxypurinol is longer than that of allopurinol. Oxypurinol shares a renal transport pathway with uric acid and ingestion of bovine milk increases the urinary excretion of uric acid. Therefore, we investigated whether its ingestion promotes the urinary excretion of oxypurinol. SUBJECTS/METHODS: Bovine milk (15 ml/kg body weight) was administered to 6 healthy subjects who took allopurinol (300 mg) 12 h prior to ingestion. In addition, a control experiment was performed with the same subjects using the same protocol, except for the ingestion of water instead of bovine milk. Blood and urine samples were collected before and after bovine and water ingestion. RESULTS: In the bovine milk ingestion experiment, the urinary excretion values of oxypurinol and uric acid were increased by 18% and 38%, respectively, and the fractional excretion values of oxypurinol and uric acid were increased by 20% and 40%, respectively, whereas those did not change in the control experiment. In addition, the concentration of alanine and sum of concentrations of amino acids were increased by 16% and 20%, respectively, in the bovine milk ingestion experiment. CONCLUSION: These results suggest that bovine milk ingestion promotes the urinary excretion of oxypurinol as well as uric acid by increasing amino acid concentration.

Endogenous endothelin-1 mediates cardiac hypertrophy and switching of myosin heavy chain gene expression in rat ventricular myocardium.

OBJECTIVES: We investigated the role of endogenous endothelin-1 in the development of cardiac hypertrophy in vivo under pressure overload conditions. BACKGROUND: Endothelin-1, a potent vasoconstrictor peptide, has recently been shown to act as a growth factor of myocardial cells in culture. METHODS: We examined the effect of an endothelin-A receptor antagonist (FR139317) on the development of right ventricular hypertrophy in rats with monocrotaline-induced pulmonary hypertension. Three groups of rats were studied: those given monocrotaline alone or monocrotaline plus FR139317 and those given vehicle alone (control group). RESULTS: The ratio of right ventricular systolic pressure to aortic systolic pressure was similarly elevated in rats treated with monocrotaline and monocrotaline plus FR139317. The right ventricular/left ventricular weight ratio was increased in monocrotaline-treated rats but lower in rats treated with monocrotaline plus FR139317 than in those treated with monocrotaline alone (p < 0.01). As a biochemical marker of hypertrophy, the isoform ratio of beta-myosin heavy chain protein was determined for the right ventricular tissue samples. This ratio was increased in all monocrotaline-treated rats but was lower (p < 0.01) in rats given monocrotaline plus FR139317 than in those given monocrotaline alone. The isoform ratio of beta-myosin heavy chain messenger ribonucleic acid quantitated by S1 nuclease mapping also was lower (p < 0.025) in rats receiving monocrotaline plus FR139317 than in those receiving monocrotaline alone. CONCLUSIONS: These data suggest that blocking the action of endothelin-1 with a receptor antagonist ameliorates cardiac hypertrophy in this model system, and that this action is not mediated by ameliorating hemodynamic changes.

Fungus invasion into human hair tissue in black dot ringworm: light and electron microscopic study.

In order to investigate the morphology of fungi invading into the human hair tissue, three cases of black dot ringworm caused by Trichophyton violaceum and Trichophyton glabrum were studied by light and electron microscopy. Fungal elements were mainly present in the hair cortex and showed a constant morphologic change during the differentiation of hair layers. The fungal elements, located deep in the keratogenous zone of the cortex, showed less electron dense non-septate hyphae. Distally, the hyphae showed septation and contained several scattered dense bodies in the cytoplasm. At the level where the Huxley's layer was keratinized, the fungal elements were transformed into arthrospores, which occupied the large volume of the cortex; each spore was surrounded by a fiber- and melanosome-free, electron lucent halo. Fungal elements occasionally invaded the keratinized hair cuticle and keratinized inner root sheath in a few hair follicles. Fungi do not invade the hair germinative cells. There seems to be a distinct relationship between the morphology of the invading fungi and the cortical cell differentiation in black dot ringworm; a balance between the fungus proliferation and the cortical cell development may be present.

Hypothalamic control of pituitary and adrenal hormones during hypothermia.

In order to investigate neuroendocrinological mechanisms of hypothermia, we determined the changes in plasma concentrations of corticosterone (CS), prolactin (PRL), and thyrotropin (TSH), and their correlations with alterations in hypothalamic dopamine (DA) and thyrotropin releasing hormone (TRH), in rats restrained and immersed in a water bath at various temperatures. A graded decrease of body temperature induced a progressive increase in the plasma level of CS, whereas that of PRL showed a drastic decrease. The plasma level of TSH also showed an increase during mild hypothermia (about 35 degrees C), but this increase was not evident during profound hypothermia (below 24 degrees C). The changes in these hormones were readily reversed by rewarming animals. Although DA content in the hypothalamus was not affected, its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), showed an increase following the decrease of body temperature. Pretreatment of the animals with sulpiride, a D2-antagonist, prevented the hypothermia-induced inhibition of PRL release. Hypothalamic TRH was significantly decreased during mild hypothermia, and it returned to control levels after rewarming. These results suggest that the decrease in plasma PRL induced by hypothermia may be associated with the activation of hypothalamic DA neurons, whereas the increase in plasma TSH during mild hypothermia seems to be caused by the increased release of TRH in the hypothalamus.

The effects of volatile anesthetics on the binding of 1-anilino-8-naphthalene sulphonate to biological membranes and lipid vesicles: the role of cholesterol.

The effects of volatile anesthetics on the properties of membrane surfaces were studied using the negatively charged fluorescent probe 1-anilino-8-naphthalene sulphonate (ANS). Although the addition of the anesthetics caused no change of the fluorescence quantum yield of ANS bound to the biological membranes, there was a significant increase of the number of binding sites (n) for ANS to the membranes. This increase was also found after treating the membranes with either trypsin or neuraminidase and with vesicles of total lipids extracted from erythrocytes. With phosphatidylcholine (PC) vesicles, the fluorescence increase by the addition of anesthetics was observed only when the vesicle contained cholesterol. The greater increase of the n value was seen in vesicles containing a higher concentration of cholesterol. When the neutral probe, N-phenyl-1-naphthylamine (NPN) was used instead of ANS, this fluorescence increase was not seen. These results were interpreted in terms of the electrostatical change of the membrane lipid region. That is, a change of the surface potential of the membrane is possibly caused by the anesthetics through the interaction with lipid components. Cholesterol plays a critical role in this interaction.

Alterations in cerebral ?-adrenergic receptor-adenylate cyclase system induced by halothane, ketamine and ethanol.

The effect of halothane, ketamine and ethanol on ?-adrenergic receptor adenylate cyclase system was studied in the brain of rats. An anesthetic concentration of halothane and ketamine added in vitro decreased the stimulatory effect of norepinephrine on cyclic AMP formation in slices from the cerebral cortex. On the other hand, ethanol increased the basal activity of cerebral adenylate cyclase without affecting on the norepinephrine-stimulated activity. The increase of the basal activity induced by ethanol was not antagonized by propranolol, a ?-adrenergic antagonist. In the crude synaptosomal (P(2)) fraction, these drugs had no significant effect on the basal adenylate cyclase activity, binding of [(3)H]dihydroalprenolol to ?-receptor, and binding of [(3)H]guanylylimido diphosphate ([(3)H]Gpp(NH)p) to guanyl nucleotide binding site. In contrast, the adenylate cyclase activity stimulated by Gpp(NH)p or NaF was significantly inhibited by an anesthetic concentration of these drugs. An anesthetic concentration of these drugs increased the membrane fluidity of P(2) fraction monitored by the fluorescence polarization technique. The addition of linoleic acid (more than 500 ?M) also induced not only the increase of fluidity, but also the decrease of Gpp(NH)p- or NaF-stimulated adenylate cyclase activity in the cerebral P(2) fraction. The present results suggest that general anesthetics may interfere with the guanyl nucleotide binding regulatory protein-mediated activation of cerebral adenylate cyclase by disturbing the lipid region of synaptic membrane.

Involvement of endogenous thyrotropin-releasing hormone in central regulation of the cardiovascular system after bleeding in conscious rats.

The 4th ventricle of a conscious rat was perfused using a push-pull cannula. The concentration of thyrotropin-releasing hormone (TRH) in the perfusate was significantly increased after withdrawal of 30% of the total blood. Administration of antiserum of TRH into the ventricle potentiated and prolonged the hypotension induced by the bleeding. These results suggest that endogenous brain TRH is involved in the central regulation of the cardiovascular system after bleeding in conscious rats.

Immunohistochemical demonstration of simple epithelia-type keratin intermediate filament in a case of Merkel cell carcinoma.

A case of Merkel cell carcinoma that developed on the right cheek of a 77-year-old woman is reported. The diagnosis was ultrastructurally made by demonstrating dense-core granules in the cytoplasm of the tumor cells. Immunohistochemically, the tumor cells were shown to possess simple epithelia-type keratin intermediate filaments, but no neurofilaments. This finding was in accordance with that obtained on normal Merkel cells. No bioactive peptides examined could be detected in the tumor cells. Typing of intermediate filaments in tumor cells may be one of the important markers, along with the ultrastructural findings, in diagnosing Merkel cell carcinoma.

Epidermolysis bullosa simplex (Koebner) is a keratin disorder. Ultrastructural and immunohistochemical study.

A skin biopsy specimen was obtained from a 1-month-old female with epidermolysis bullosa simplex (Koebner). Histologically, an intraepidermal separation was seen and considered to be formed by cytolysis of the epidermal basal cells. Ultrastructurally, the basal cells were lacking in cytoplasmic tonofilaments, and the initial change of the cytolysis seemed to be cleavages of the cytoplasm. Immunohistochemically, a basal cell keratin was expressed in a suprabasal cell layer but not in the basal cell layer, and a panepithelial keratin was not detected in the basal cell layer. These findings suggest that keratin production of the epidermal cells may be delayed, resulting in a weakness of the basal cells against minor trauma to the skin.

An antiserum against amyloid beta-protein precursor detects a unique peptide in Alzheimer brain.

An antiserum was raised against an amino acid sequence predicted from the DNA sequence of amyloid beta-protein precursor (ABPP), and it was then affinity-purified. This affinity-purified antibody (anti-GID) intensely stained neurons and dystrophic neurites in plaques of Alzheimer's disease (AD) patients, but marginally stained neurons of age-matched normal individuals. Anti-GID antibody detected a series of protein bands with a molecular weight centered at 100,000 and a second band at 55,000 on a blot of the human brain particulate fraction. It also stained a set of bands with a molecular weight around 95,000 and a doublet of Mr 16,000 in the soluble fraction. A band at Mr 35,000 was detected in the soluble fraction prepared from brain tissue of AD patients but not from control brain tissue. A strong immunostaining of AD sections with anti-GID and the presence of a Mr 35,000 band unique to AD might reflect an altered processing of ABPP in AD brains.

The involvement of central cholinergic mechanisms in cardiovascular responses to intracerebroventricular and intravenous administration of thyrotropin-releasing hormone.

Intracerebroventricular (i.c.v.) administration of thyrotropin-releasing hormone (TRH) in a range from 0.1 to 100 micrograms induced a dose-related increase in blood pressure in conscious rats, whereas TRH-free acid (TRH-OH) and histidyl-proline diketopiperazine (His-Pro-DKP), metabolites of TRH, did not. The blood pressure responses to intravenous (i.v.) injection of 5 mg/Kg TRH were similar to those induced by TRH (i.c.v.). Pretreatment with atropine (50 micrograms, i.c.v.) significantly reduced the pressor effect of TRH administered through either route. Hemicholinium-3 (50 micrograms, i.c.v.), an inhibitor of choline uptake, also prevented the increase in blood pressure induced by TRH (10 micrograms, i.c.v.). These results indicate that both centrally and peripherally administered TRH have pressor effects that are mediated by central cholinergic mechanisms, probably by activating cholinergic neurons.

Cardiovascular effect of intravenously administered thyrotropin-releasing hormone and its concentration in push-pull perfusion of the fourth ventricle in conscious and pentobarbital-anesthetized rats.

Changes in the concentration of thyrotropin-releasing hormone (TRH) in cerebrospinal fluid (CSF) were examined by the push-pull perfusion method after intravenous (i.v.) administration of the peptide in conscious and pentobarbital-anesthetized rats. The concentration of endogenous TRH in the perfusate was not changed during the 160-min perfusion period and was similar to that in the CSF (0.92 +/- 0.26 ng/ml) collected before the perfusion in conscious as well as in anesthetized rats. After i.v. administration of TRH (5 mg/kg) to the conscious rats, the peptide concentration in the perfusate increased to 42.23 +/- 14.33 ng/ml during the first 20 min and gradually returned to the basal level 2 hr after administration. The total amount of TRH detected in the perfusate was 20.0 ng. It was reduced by 75% in the anesthetized animals. The increases in blood pressure and heart rate, seen after i.v. as well as intracerebroventricular administration of TRH in the conscious rats, was significantly inhibited in the anesthetized rats. These results indicate that systemically administered TRH exerts its cardiovascular effect at central site(s), and that the transportation and the effect of the peptide is suppressed by pentobarbital anesthesia.

Hemorrhage-induced regional brain thyrotropin-releasing hormone release in conscious rats measured by microdialysis.

The septum, nucleus accumbens and preoptic area in the brains of conscious, freely moving rats were perfused using microdialysis probes. The TRH concentration significantly increased in the septum after withdrawal of 30% of the total blood volume but remained at constant levels in the other brain areas. Also, high potassium dose-dependently stimulated TRH release in vivo. These results suggest that blood loss stimulates septal TRH release, probably by membrane depolarization of TRH-containing nerve terminals.

Trichophyton rubrum invasion of human hair apparatus in tinea capitis and tinea barbae: light- and electron microscopic study.

We have previously reported morphological changes of Trichophyton violaceum and Microsporum canis in hair apparatuses in tinea capitis. To investigate the morphology of Trichophyton rubrum in the human hair apparatus, two cases of tinea capitis and one case of tinea barbae were examined by light- and electron microscopy. The fungal elements, which were located in the lower keratogenous zone, showed non-septate hyphae in the outer part of the hair cortex. With the upward development of the hair layers, some hyphae invaded the keratinized hair cuticle and keratinized inner root sheath and were transformed into arthrospores. Some hyphae remaining in the hair cortex were also transformed into arthrospores, while other hyphae in the hair cortex did not survive, but degenerated. In T. rubrum hair infection, there is a distinct relationship between the morphological changes of the fungi and the hair cell differentiation as seen in T. violaceum and M. canis infections. However, T. rubrum displays unique morphological changes, which are different from those of T. violaceum and M. canis, in hair apparatuses.

Relationship between tissue reactions and morphological changes of the fungi in chromoblastomycosis: morphometry and electron microscopy.

To investigate the histological distribution and the morphology of the fungi and the tissue reactions in chromoblastomycosis, especially in the process of transepidermal elimination, cutaneous lesions of two patients with this disease were studied morphometrically and ultrastructurally. In the dermis, most of the fungal elements appeared as sclerotic cells and their cell wall showed an irregular, worm-eaten leaf-like appearance; they seemed to be continuously attacked by polymorphonuclear neutrophils. The epidermis eliminated 10-20% of all the organisms in the skin lesions, and the hypha-forming activity tended to be higher in the epidermis than in the dermis. Ultrastructurally, basal keratinocytes facing the dermal abscess containing fungal elements frequently appeared as dark cells, suggesting an increased proliferation activity. Spinous keratinocytes facing intraepidermal microabscesses containing fungal elements showed an abnormal accumulation of tonofilaments and further early keratinization in the spinous cell layer. All of the morphological changes of the dermis and epidermis are regarded as defence reactions against the fungi existing in the skin lesions. There is a close relationship between tissue reactions and morphological changes of fungi in chromoblastomycosis.

Fungus invasion of human hair tissue in tinea capitis caused by Microsporum canis: light and electron microscopic study.

Previously, we reported a morphological change of Trichophyton violaceum in hair tissue in black dot ringworm. To investigate the morphology of Microsporum canis in human hais tissue, three cases of tinea capitis by M. canis were examined by both light and electron microscopy. The fungal elements, which were located deeplyin the keratogenous zone, showed nonseptate hyphae in the outer part of the hair cortex. With the upward development of hair tissues, some hyphae invaded the keratinized inner root sheath and were there transformed into arthrospores, which then occupied the large volume of the inner root sheath; each spore was surrounded by an electron-lucent halo. In some affected hair follicles, at the follicular isthmus level, a microabscess composed of polymorphonuclear leukocytes was often formed in the outer root sheath adjacent to the arthrospores in the keratinized inner root sheath. On the other hand, the remaining hyphae in the cortex became degenerated. Fungi did not invade the hair-germinative cells. There is a distinct relationship between the morphological change of fungi and the differentiation of hair cells in tinea capitis by M. canis as well as in that by T. violaceum, although the direction of invasion and pathological roles of fungal elements within hair tissue are significantly different between the two species of fungi.

Effects of hypothermia on thyrotropin-releasing hormone content in the rat brain.

The thyrotropin-releasing hormone (TRH) content in the brain was determined in normothermic and hypothermic rats subjected to immobilization stress. TRH contents in the hypothalamus, midbrain and cerebral cortex significantly decreased during mild hypothermia (body temperature about 34 degrees C), but not during profound hypothermia (about 24 degrees C). The decreases in the TRH content during mild hypothermia were readily reversed by rewarming the animal. These results indicate that cerebral TRH is involved in the response to a mild body temperature drop when the animal is exposed to a cold environment.

Alteration of the turnover of dopamine and 5-hydroxytryptamine in rat brain associated with hypothermia.

The alteration of monoamines and their metabolites in the brain in response to hypothermia was studied using rats subjected to a cold and immobilization stress. The experiments were designed to compare the responses in the "hypothermal" rats with those in the "normothermal" ones which received the same stress except for the change in body temperature. It has been found that the contents of norepinephrine (NE) and 5-hydroxytryptamine (5-HT) in various cerebral regions were significantly decreased during hypothermia. These decreases were readily reversed by the rewarming of animals. Moreover, the increase in the content of 5-hydroxyindole-3-acetic acid (5-HIAA), the metabolite of 5-HT, was also detected in some cerebral regions where the decrease of 5-HT was observed. Although the dopamine (DA) contents in all cerebral regions examined were found to be unaltered, its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and/or homovanillic acid (HVA) contents in most regions in the brain showed a significant elevation during and/or after the occurrence of hypothermia. These results suggest that the metabolic turnovers of 5-HT and DA in various cerebral regions may be accelerated during hypothermia.

Clorgyline-induced increases in presynaptic DA: changes in the behavioral and neurochemical effects of amphetamine using in vivo microdialysis.

Microdialysis was used in behaving rats to further characterize the behavioral and regional dopamine (DA) response to the monoamine oxidase (MAO) inhibitor clorgyline and determine how MAO inhibition affects amphetamine (AMPH)-induced changes in behavioral and extracellular DA dynamics. Although clorgyline (4.0 mg/kg) did not significantly alter behavior, it produced prolonged increases in caudate and accumbens extracellular DA and 3MT and corresponding decreases in homovanillic acid (HVA) and dihydroxyphenylacetic acid (DOPAC). Clorgyline pretreatment altered the behavioral response to both low (0.25 mg/kg) and moderate (2.5 mg/kg) doses of AMPH, which was characterized by a shift to more intense forms of stereotype and corresponding decreases in locomotion. The caudate and accumbens DA response to AMPH (0.25 mg/kg) was also significantly augmented, consistent with an increase in AMPH-releasable cytoplasmic DA. Thus, the potentiated DA response in clorgyline-pretreated animals may be responsible for the changes in the stereotypy profile. Possible implications of these observations for the augmented behavioral response observed with repeated AMPH administration are discussed.