Childhood bone tuberculosis from Roman Pécs, Hungary.

A child from a Roman necropolis in Pécs, Hungary (4th century CE) was initially diagnosed with severe spinal osteomyelitis. The post-cranial skeleton displayed bone alterations in the lower thoracic and upper lumbar segments, including vertebral body destruction, collapse and sharp kyphosis, and additional multiple rib lesions, suggesting a most likely diagnosis of pulmonary and spinal tuberculosis. This study discusses a number of selected diagnoses in the context of our pathological findings, complementing the macroscopic examination with radiological and biomolecular analyses.

Amperometric cell for subcutaneous detection of hydrogen sulfide in anesthetized experimental animals.

Hydrogen sulfide (H2S) is a toxic gas. It has been recognized that H2S evolving in biochemical reactions in living organisms has an important role in different physiologic processes. Nowadays, H2S is known as an endogenous messenger molecule. Natural sulfurous spring water has been proved beneficial in the therapy of diseases of the skin and other organs (Boros et al 2013). In vivo real-time detection of local H2S concentration is an important but challenging task.We developed a two-electrode amperometric cell for selective subcutaneous detection of H2S in anesthetized mice. The cell is a small size implantable gas sensor containing a platinum disc anode and a silver cathode. The selectivity is provided by a membrane permeable only by gases. There is a buffered reversible electrochemical mediator solution in an oxidized form inside the cell. As gaseous H2S penetrates into the cell the mediator is reduced, and +0.4 V versus the reference is employed on the platinum working electrode. The reduced mediator is oxidized on the anode surface. The current provides an analytical signal representing the concentration of H2S.Appropriate shape, size and membrane material were selected, and optimal working parameters--such as mediator concentration, pH and cell voltage--were determined in vitro. The lower limit of detection in the stirred sample solution at pH = 5.5 was as small as 9.4 × 10(-7) M and a dynamic concentration range of 0-6 × 10(-4) M could be achieved.The detecting surfaces of the cell were covered with freshly dissected mouse skin to test dermal H2S permeability. In other experiments, the cell was implanted subcutaneously in an anesthetized mouse and the animal was submerged in a buffer solution containing different concentrations of H2S so that the skin surface over the sensor was covered by the solution. Measurements of subcutaneous H2S concentration were taken. The experiments clearly proved that H2S diffuses through the skin of the live mouse.

Chronomics: circadian lead of extrapineal vs. pineal melatonin rhythms with an infradian hypothalamic exploration.

A circadian rhythm is documented for plasma, pineal, and hypothalamic melatonin of male and female rats kept on staggered lighting regimens. Log[_10]-transformation of the data usually normalizes, when need be, the distribution of residuals from the 24-hour cosine curve fits. A tentative circadian acrophase chart is presented that shows a lead in circadian acrophase of duodenal over pineal melatonin. The use of antiphasic lighting regimens facilitates circadian studies that can be carried out for several days, thereby allowing the assessment of infradian components such as a circasemiseptan variation in hypothalamic melatonin documented herein. The results are qualified by the presence of a second extremum of a double magnetic storm at the start of mapping.

Chronomics, neuroendocrine feedsidewards and the recording and consulting of nowcasts--forecasts of geomagnetics.

A multi-center four-hourly sampling of many tissues for 7 days (00:00 on April 5-20:00 to April 11, 2004), on rats standardized for 1 month in two rooms on antiphasic lighting regimens happened to start on the day after the second extremum of a moderate double magnetic storm gauged by the planetary geomagnetic Kp index (which at each extremum reached 6.3 international [arbitrary] units) and by an equatorial index Dst falling to -112 and -81 nT, respectively, the latter on the first day of the sampling. Neuroendocrine chronomes (specifically circadian time structures) differed during magnetically affected and quiet days. The circadian melatonin rhythm had a lower MESOR and lower circadian amplitude and tended to advance in acrophase, while the MESOR and amplitude of the hypothalamic circadian melatonin rhythm were higher during the days with the storm. The circadian parameters of circulating corticosterone were more labile during the days including the storm than during the last three quiet days. Feedsidewards within the pineal-hypothalamic-adrenocortical network constitute a mechanism underlying physiological and probably also pathological associations of the brain and heart with magnetic storms. Investigators in many fields can gain from at least recording calendar dates in any publication so that freely available information on geomagnetic, solar and other physical environmental activity can be looked up. In planning studies and before starting, one may gain from consulting forecasts and the highly reliable nowcasts, respectively.

Galactoside-specific misletoe lectin modulates dexamethasone-induced apoptosis and glucocorticoid receptor level in Balb/c thymocytes.

The galactoside-specific plant lectin, Viscum album agglutinin-(VAA)-I has been shown to activate the natural immune system and modulate the maturation of thymocytes in vivo. However the mechanism of this immunobiological action is not yet understood. In our previous study we demonstrated the VAA-I-induced enhancement of proliferation and selection of thymocytes which inhibited the dexamethasone (DX)-induced thymocyte depletion. In this present work we investigated the effect of 1, 4 and 21 days of VAA-I treatment on DX-induced apoptosis of thymocytes in Balb/c mice. The number of early apoptotic cells was detected with Annexin V staining while the late apoptotic cells were identified according to their propidium iodide incorporation into DNA using flow cytometry. The expression of glucocorticoid receptor (GCR) in double-negative (DN), double-positive (DP) and CD4 or CD8 single-positive (SP) cell populations was assessed. The additive effect of lectin on DX-induced apoptosis of thymocytes consisted of two different actions of VAA-I and DX. One-day VAA-I treatment caused enhanced apoptosis in SP mature cells in contrast to the apoptotic effect of DX, which was mainly directed towards immature DN and DP cells. Treatment with 30 ng/kg VAA-I for four days elevated the GCR level (mean fluorescence intensity) in DP thymocytes. Lectin treatment for 21 days caused more than 20% elevation of GCR expression in all thymocyte subpopulations (DN, DP, CD4+ and CD8+). These results suggest that VAA-I may alter the sensitivity of thymocytes to glucocorticoids and this effect may play a role in the bell-shaped dose-response curve of lectin-induced immunological effects.

Exercise and vitamin E intake are independently associated with metabolic abnormalities in human immunodeficiency virus-positive subjects: a cross-sectional study.

We investigated the relationship among habitual exercise, diet, and the presence of metabolic abnormalities (body fat redistribution, dyslipidemia, and insulin resistance) in a cross-sectional study of 120 human immunodeficiency virus (HIV)-infected subjects with use of bivariate and multivariate regression-analysis models. Total and aerobic exercise were significantly and negatively associated with fasting plasma triglyceride levels in the entire sample and in the fat redistribution group. Inverse associations between total or aerobic exercise and insulin resistance were suggestive but did not achieve statistical significance. Diastolic blood pressure was significantly and inversely associated with supplemental or total but not habitual dietary intake of vitamin E. In conclusion, exercise and vitamin E intake were independently and negatively associated with several phenotypic manifestations of HIV-associated metabolic syndrome, whereas other macro- or micronutrients did not have comparable significance.

Brain structures mediating the suckling stimulus-induced release of prolactin.

Suckling-induced prolactin release is a widely studied neuroendocrine reflex, comprising a neural afferent and a humoral efferent component. The information on the brain structures involved in this reflex is fairly limited. The present studies focused on this question. The following hypothalamic interventions were made in lactating rats and the dams were tested for the suckling-induced prolactin response: (i) unilateral or (ii) bilateral frontal cuts at the level of the anterior and posterior hypothalamus; (iii) administration of 5,7-dihydroxytryptamine or (iv) 6-hydroxydopamine into the hypothalamic paraventricular nucleus (PVN) to destroy serotonergic and catecholaminergic innervation of the cell group, respectively; (v) lesion of the medial subdivision of the PVN; and (vi) horizontal knife cuts below the PVN. Bilateral posterior and bilateral or unilateral anterior frontal cuts caused blockade of the suckling-induced release of prolactin. Likewise, most dams receiving 5,7-dihydroxytryptamine in the PVN did not respond to the suckling stimulus. Immunocytochemistry revealed that, in those rats which did not show a rise in plasma prolactin, there were almost no serotonergic fibres and terminals in the PVN, while in dams which exhibited a response, numerous serotonergic elements were evident. 6-Hydroxydopamine treatment did not cause significant alteration in the prolactin response. Lesion of the medial, largely parvocellular subdivision of the PVN, or horizontal knife cuts below this cell group, blocked the hormone response. The findings demonstrate for the first time that: (i) interruption of the connections between the brain stem and the hypothalamus interferes with the prolactin response to the suckling stimulus; (ii) serotonergic fibres terminating in the hypothalamic PVN are involved in the mediation of the suckling stimulus; and (iii) within the PVN, neurones in the medial, largely parvocellular subdivision of the cell group take part in the transfer of the neural signal, eventually inducing prolactin release.

Active water selective channels in the stomach: investigation of aquaporins after ethanol and capsaicin treatment in rats.

UNLABELLED: Recent studies discovered the existence of aquaporins (AQP), suggesting their roles in the active, ATP dependent water secretion or absorption. Our recent development of the monoclonal antibody family against aquaporins (Type 1 and 4) allowed us a good opportunity to investigate the mechanism of the gastric mucosal edema in a rat model. THE AIM OF OUR STUDY was to evaluate the changes in the tissue level of aquaporins (AQP1 and AQP4) after ethanol and capsaicin treatment in rat stomach. MATERIALS AND METHODS: the experiments were carried out on Sprague-Dawley rats weighing 150-200 g. The animals were fasted for 24 h, after the 1 ml of ethanol (50% v/v) or capsaicin (2 mg/ml) was given intragastrically. Rats were sacrificed after 5, 30, 60, 120 and 240 min, the tissue level of AQP1 and AQP4 was investigated immunoserologically by ELISA and dot-blot methods using our monoclonal antibodies. The location of these aquaporins in the gastric tissue was demonstrated by immunohistochemistry. RESULTS: (1) in ethanol-treated stomach, both AQP1 and AQP4 increased after 5 min simultaneously with gastritis, then decreased dramatically depending on time. (2) In the capsaicin-treated group there were no changes in the tissue level of aquaporins in the first hour. After 60 min both AQP1 and AQP4 increased in the stomach without any macroscopically detectable changes, then decreased depending on time. (3) The immunohistochemical investigations using our monoclonal antibodies seem to support our present quantitative results. CONCLUSION: chemically induced gastric mucosal lesions are started by an extended edema. In the induction of the edema and the subsequent gastric injury, aquaporins (both AQP1 and AQP4) play an important role in the maintenance of mucosal integrity.

Immunoneutralization of prolactin prevents stimulatory feedback of prolactin on hypothalamic neuroendocrine dopaminergic neurons.

We have found that exogenous prolactin (PRL) stimulates all three populations of hypothalamic neuroendocrine dopaminergic neurons. In this study, we investigated the effects of immunoneutralization of endogenous PRL on the activity of these neurons. Injection of 17beta-estradiol (E2) (20 microg subcutaneously) 10 d after ovariectomy induced a proestrus-like increase in PRL in peripheral plasma the following afternoon. At 1000 h the day after E2 injection, rats received either rabbit antirat PRL antiserum (PRL-AS) (200 microL) or normal rabbit serum (NRS, 200 microL, controls) intraperitoneally. Groups of rats were then decapitated every 2 h from 1100 h to 2100 h. Trunk blood was collected and serum extracted with protein A to remove the PRL-AS/PRL complex, and the remaining free PRL was measured by radioimmunoassay. Sites of neuroendocrine dopaminergic nerve terminals, the median eminence (ME), and intermediate and neural lobes of the pituitary gland were excised and stored for determination of dopamine (DA) and 3,4-dihydroxyphenyl acetic acid (DOPAC) concentrations by high-performance liquid chromatography electrochemical detection (EC). In addition, the anterior lobe of the pituitary gland, the locus of DA action, was collected. The concentration of PRL in NRS-treated animals increased by 1500 h, peaked by 1700 h, and returned to low levels by 2100 h. PRL-AS prevented the increase in PRL secretion in response to E2. The turnover of DA (DOPAC:DA ratio; an index of dopaminergic neuronal activity) in the ME of NRS-treated animals increased at 1500 h and rapidly returned to basal levels. Treatment with PRL-AS prevented the increase in DA turnover in the ME. DA turnover in the intermediate lobe increased coincident with the peak of PRL in serum of NRS-treated rats. PRL-AS administration prevented increased DA turnover in the intermediate lobe. The turnover of DA in the neural lobe increased by 1300 h and decreased steadily through 2100 h. However, administration of PRL-AS minimally suppressed the turnover of DA in the neural lobe. Moreover, administration of PRL-AS attenuated the rise of DA in the anterior lobe associated with the waning phase of the E2-induced PRL surge. These results clearly indicate that endogenous PRL regulates its own secretion by activating hypothalamic neuroendocrine dopaminergic neurons.

A molecular recognition hypothesis for nonpeptides: Na+ K+ ATPase and endogenous digitalis-like peptides.

The molecular recognition hypothesis for peptides is that binding sites of ligands and their receptors are encoded by short, complementary segments of DNA. A corollary hypothesis for nonpeptide ligands posited here is that peptide replicas may be encoded by the DNA segment complementary to the receptor binding sites for nonpeptides. This corollary was tested for digitalis. a family of cardiotonic and natriuretic steroids including ouabain. A hexapeptide (ouabain-like peptide, OLP) complementary to a ouabain binding site on sodium potassium dependent adenosine triphosphatase (Na+ K+ ATPase) exhibited activity in a digitalis bioassay. Antisera to the complementary peptide (OLP) stained the neurohypophysis in an immunocytochemical procedure. The complementary peptide was found to share an identical 4-amino acid region with the 39-amino acid glycopeptide moiety of the vasopressin-neurophysin precursor. This glycopeptide was isolated from pituitary extracts; it exhibited digitalis-like activity in the submicromolar range and cross-reacted with complementary peptide antibodies. Another digitalis-like substance with high activity also was detected in the extracts. These results demonstrate that the vasopressin-neurophysin glycopeptide has digitalis-like activity. Moreover, the findings are consistent with the hypothesis that peptide mimetics of nonpeptides are encoded in the genome.

The effect of neurointermediate lobe denervation on hypothalamic neuroendocrine dopaminergic neurons.

The contribution of tuberohypophyseal and periventricular-hypophyseal dopaminergic neurons to the regulation of the secretion of prolactin (PRL) has yet to be clarified. In this study, we used pituitary stalk compression to disrupt hypothalamic neural input to the neurointermediate lobe (NIL). Neurointermediate lobe denervation (NIL-D) selectively disrupts the axons of tuberohypophyseal and periventricular-hypophyseal dopaminergic neurons, while leaving tuberoinfundibular dopaminergic neurons and the vascular supply of the pituitary gland intact. NIL-D was performed in ovariectomized (OVX) rats. The concentration of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME) and various regions of the pituitary gland of OVX and OVX+NIL-D rats were measured by HPLC-EC. The concentration of PRL, alpha-melanocyte stimulating hormone (alpha-MSH), and luteinizing hormone (LH) in serum were determined by radioimmunoassay. Successful NIL-D was confirmed by increased water intake. One week after NIL-D, serum PRL and alpha-MSH were elevated, but there was no change in the concentration of LH in serum. The concentration of DA was increased in the median eminence (ME), decreased in the outer zone of the anterior lobe (AL-OZ), as well as the intermediate (IL) and neural lobes (NL), and remained unchanged in the inner zone of the anterior lobe (AL-IZ). The concentration of DOPAC was increased in the ME and NL, decreased in the IL, and remained unchanged in both the AL-IZ and AL-OZ. These data confirm that pituitary stalk compression denervates the NIL. Moreover, decreases in the concentration of DA in the IL and AL-OZ, coupled with elevation of serum PRL and alpha-MSH indicate that DA from the NIL contributes to the increased inhibition of the secretion of PRL and alpha-MSH in OVX rats.

New combination for the therapy of canine otitis externa. II. Efficacy in vitro and in vivo.

A new, effective drug combination was developed for the therapy of canine otitis externa (OE) by taking into consideration the microorganisms most frequently isolated from dogs affected with OE, their combination, their drug sensitivity and the type and incidence of ear canal inflammation. The antimycotic active ingredient of the combination is ketoconazole, its antibacterial component is gentamicin sulphate, while its antiphlogistic constituent is mazipredone hydrochloride. Based upon the results of in vitro pharmacodynamic tests, the antifungal activity of the combination is superior to that of ketoconazole used alone at the same concentration. A total of 210 dogs affected with OE were treated with the combination: 94.2 per cent of them became clinically symptomless and microbiologically negative in an average of 8.5 days. No adverse reactions were observed in connection with the use of the drug combination. The therapeutic results can be attributed to the high antifungal efficacy of the combination demonstrated in vitro and to the favourable properties of the solvent mixture.

[Study of the cost-effectiveness of sequential antibiotic (ciprofloxacin) therapy]. / Szekvenciális antibiotikum (ciprofloxacin) terápia hatékonyságának és költségének összevetése.

A multicenter, prospective, randomized study was performed to compare the cost-effectiveness of parenteral antibiotic versus parenteral antibiotic + oral ciprofloxacin sequential therapy in the treatment of moderate and severe infections in hospitalized patients. After deciding on inclusion (n = 298), the patients were randomly allocated in group "A" (evaluable 120 cases) receiving only parenteral antibiotics, or in group "B" receiving parenteral antibiotic + oral ciprofloxacin from the 3.-5. day of therapy on. The local physician decided on the starting parenteral therapy and the necessity of other therapeutic interventions, and in case of group "B" patients on the possibility of switching parenteral to oral therapy. The overall cure rate was 68% in group "A" and 82% in group "B" (improved 33 and 21, failure 5 and 1 cases resp.). Of the direct costs in patients, care, the costs of antibiotic treatments based on purchase prices was compared: the cost of antibiotic treatment in group "B" was 32% less than in group "A", and the sequential therapy saved 9000 Ft/patient in average. In conclusion, ciprofloxacin proved to be an effective antimicrobial agent that can also be used in sequential therapy and the oral administration resulted in a significant save of money with the same therapeutic effectiveness.

Electrical potential dissipation induced by free fatty acids in pea stem mitochondria.

Linolenic, linoleic, oleic, palmitic and stearic acids (FFA) collapse the electrical potential of pea stem mitochondria in the absence or in the presence of 0.5 mM Mg2+. Higher concentrations of this cation (5 mM) lower the rate of dissipation caused by linoleic, oleic and palmitic acids, while abolishing that induced by stearic acid. Carboxyatractyloside and ADP do not reverse the FFA-induced collapse both in the presence or absence of Mg2+. EDTA, EGTA or BHT do not influence the dissipation caused by FFA that, in addition, is not linked to lipid peroxidation evaluated as malondialdehyde or conjugated diene formation. Only linolenic acid sustains a peroxidation which, however, appears to be caused by its own oxidation catalysed by lipoxygenases rather than by membrane lipoperoxidation induced by this free fatty acid. These results suggest that neither the ATP/ADP exchanger nor lipid peroxidation appear to be involved in FFA-induced uncoupling in pea stem mitochondria.

Absorption of colostral immunoglobulins in suckling piglets.

The three portions of the small intestine of 18 neonatal piglets were examined by using the so-called scroll technique evolved by the authors and in cross sections. The chemical constitution and the immunoglobulin content of undigested protein droplets being under absorption were studied. An intensive absorption of immunoglobulins was demonstrated in the body of each cuticulated cylindric epithelial cell of the mucosa covering the jejunum and the proximal two-third of the ileum. Absorption was the most intensive, though variable by area, in the jejunum. It was demonstrable 4 hr after birth and reached the highest intensity between 8 and 12 hr, then tended to decline. In the protein droplets alpha-amino acids, neutral mucopolysaccharides and protein-bound Ca2+ were detected. Alkaline phosphatase activity was demonstrated around the droplets. Immunofluorescence and immunoperoxidase reactions proved the presence of immunoglobulins, first of all of IgG, in droplets. Electron microscopically, the microvilli of the involved epithelial cells were swollen, sometimes neighbouring ones were pushed away from each other; in the same areas, an electron-dense substances (colostrum) covering the enterocytes was streaming through the tubulovascular network of cells to the perinuclear region, where it appeared in spherical droplets.

Ion content of synaptic vesicles.

Proton induced X-ray emission analysis measurements were performed to determine the P, S, K, Ca Fe, Ni, Cu and Zn ion content of presynaptic vesicles prepared from guinea-pig brain cortex. The number of different ions per single vesicle is calculated using the results of the additional protein content determinations. The ion contents of cholinergic and adrenergic vesicles are compared.