Co-localisation of kisspeptin with galanin or neurokinin B in afferents to mouse GnRH neurones.

The gonadotrophin-releasing hormone (GnRH) secreting neurones, which form the final common pathway for the central regulation of reproduction, are directly targeted by kisspeptin (KP) via the G protein-coupled receptor, GPR54. In these multiple labelling studies, we used ovariectomised mice treated with 17ß-oestradiol (OVX + E(2)) or vehicle (OVX + oil) to determine: (i) the ultrastructural characteristics of KP-immunoreactive (IR) afferents to GnRH neurones; (ii) their galanin or neurokinin B (NKB) content; and (iii) the co-expression of galanin or NKB with KP in the two major subpopulations of KP neurones located in the rostral periventricular area of the third ventricle (RP3V) and the arcuate nucleus (Arc). Electron microscopic investigation of the neuronal juxtapositions revealed axosomatic and axodendritic synapses; these showed symmetrical or asymmetrical characteristics, suggesting a phenotypic diversity of KP afferents. Heterogeneity of afferents was also demonstrated by differential co-expression of neuropeptides; in OVX + E(2) mice, KP afferents to GnRH neurones showed galanin-immunoreactivity with an incidence of 22.50 ± 2.41% and NKB-immunoreactivity with an incidence of 5.61 ± 2.57%. In OVX + oil animals, galanin-immunoreactivity in the KP afferents showed a major reduction, appearing in only 5.78 ± 1.57%. Analysis for co-localisation of galanin or NKB with KP was extended to the perikaryal level in animal models, which showed the highest KP incidence; these were OVX + E(2) females for the RP3V and OVX + oil females for the ARC. In the RP3V of colchicine-treated OVX + E(2) animals, 87.84 ± 2.65% of KP-IR neurones were galanin positive. In the Arc of the colchicine-treated OVX + oil animals, galanin immunoreactivity was detected in only 12.50 ± 1.92% of the KP expressing neurones. By contrast, the incidence of co-localisation with NKB in the Arc of those animals was 98.09 ± 1.30%. In situ hybridisation histochemistry of sections from OVX + E(2) animals identified galanin message in more than a third of the KP neurones in the RP3V (38.67 ± 11.57%) and in the Arc (42.50 ± 12.52%). These data suggest that GnRH neurones are innervated by chemically heterogeneous KP cell populations, with a small proportion deriving from the Arc group. The presence of galanin within KP axons innervating GnRH neurones and the oestrogen-dependent regulation of that presence add a new dimension to the roles played by galanin in the central regulation of reproduction.

Evidence for suprachiasmatic vasopressin neurones innervating kisspeptin neurones in the rostral periventricular area of the mouse brain: regulation by oestrogen.

In rodents, a circadian signal from the suprachiasmatic nucleus (SCN) is essential for the pro-oestrous surge of gonadotrophin-releasing hormone (GnRH), which, in turn, induces luteinising hormone (LH) surge and ovulation. We hypothesised that kisspeptin (KP) neurones in the anteroventral periventricular and periventricular preoptic nuclei (AVPV/PeN) form part of the communication pathway between the SCN and GnRH neurones. In anterograde track tracing studies, we first identified vasopressin (VP)-containing axons of SCN origin in apposition to KP-immunoreactive (IR) neurones. Studies to quantify this input relied on the observation that VP-synthesising neurones in the SCN differ from other VP systems in their lack of galanin expression. In ovariectomised mice, 30.79 +/- 1.63% of KP-IR perikarya and proximal dendrites within the AVPV/PeN received galanin-negative VP-IR varicosities. Oestrogen-treatment significantly increased the number of KP-IR neurones, with their percentage apposed by galanin-negative VP-IR varicosities (46.95 +/- 1.88%) and the number of VP-IR appositions on individual KP-IR neurones. At the ultrastructural level, the VP-IR terminals formed symmetric synapses with KP-IR neurones, which was in accordance with the morphology of inhibitory synapses established by SCN neurones. By contrast to VP, vasoactive intestinal polypeptide (VIP), which is synthesised by a distinct subset of SCN neurones, occurred only rarely in axons apposed to KP-IR neurones. Altogether, our results are consistent with the hypothesis that KP neurones located in the mouse AVPV/PeN receive circadian information from the SCN via a vasopressinergic monosynaptic pathway, which is enhanced by oestrogen.

Glutamatergic innervation of the hypothalamic median eminence and posterior pituitary of the rat.

Recent studies have localized the glutamatergic cell marker type-2 vesicular glutamate transporter (VGLUT2) to distinct peptidergic neurosecretory systems that regulate hypophysial functions in rats. The present studies were aimed to map the neuronal sources of VGLUT2 in the median eminence and the posterior pituitary, the main terminal fields of hypothalamic neurosecretory neurons. Neurons innervating these regions were identified by the uptake of the retrograde tract-tracer Fluoro-Gold (FG) from the systemic circulation, whereas glutamatergic perikarya of the hypothalamus were visualized via the radioisotopic in situ hybridization detection of VGLUT2 mRNA. The results of dual-labeling studies established that the majority of neurons accumulating FG and also expressing VGLUT2 mRNA were located within the paraventricular, periventricular and supraoptic nuclei and around the organum vasculosum of the lamina terminalis and the preoptic area. In contrast, only few FG-accumulating cells exhibited VGLUT2 mRNA signal in the arcuate nucleus. Dual-label immunofluorescent studies of the median eminence and posterior pituitary to determine the subcellular location of VGLUT2, revealed the association of VGLUT2 immunoreactivity with SV2 protein, a marker for small clear vesicles in neurosecretory endings. Electron microscopic studies using pre-embedding colloidal gold labeling confirmed the localization of VGLUT2 in small clear synaptic vesicles. These data suggest that neurosecretory neurons located mainly within the paraventricular, anterior periventricular and supraoptic nuclei and around the organum vasculosum of the lamina terminalis and the preoptic area secrete glutamate into the fenestrated vessels of the median eminence and posterior pituitary. The functional aspects of the putative neuropeptide/glutamate co-release from neuroendocrine terminals remain to be elucidated.

The change of attitude of Hungarian physicians towards the importance of risk factors of coronary heart disease over the period 1985-1996.

Cardiovascular mortality in Hungary is still increasing, while it shows a continual decrease in the developed Western world. The authors examined, by means of a questionnaire, the attitude of physicians, in a large county hospital, to prevention of cardiovascular diseases and promotion of a healthy way of life. The questionnaire was answered by 170 physicians, 107 (63%) males and 63 (37%) females. Eighty-six percent of them believed coronary heart disease to be preventable. Twenty-six percent of the physicians currently smoked, and 53% did not know their own cholesterol level. As a cardiovascular mortality risk factor smoking was considered the most important risk factor, with sedentary lifestyle the second, high cholesterol level the third, and hypertension being only the fourth. Hungarian hospital physicians' rating of the effect of reducing the risk factors for coronary heart disease was similar to those results published in 1986 of American doctors, there being no significant difference in the importance attributed to smoking and elevated blood cholesterol. American doctors believed that hypertension had a more important effect on coronary heart disease than did Hungarian physicians, whilst the Hungarians attributed greater importance to a diet high in fat, being overweight, having a sedentary life-style, stress, elevated triglyceride level and type A behaviour. The results of this present study which related to the doctors attitudes towards health education for their patients were compared to results obtained from a study relating to physicians in the same hospital in 1985. Only in two aspects was a significant change observed. According to the authors' opinion greater efforts should be made regarding physician education on the subject of disease prevention. Additionally the employment of well educated nurses with specific training in preventive medicine could improve the effectiveness of the prevention of coronary heart disease.

[Effect of enalapril on restrictive type diastolic filling disorder in severe congestive heart failure]. / Enalapril hatása restriktív típusú diastolés telödési zavarral járó súlyos szívelégtelenségben.

Authors investigated the effect of enalapril (10-20 mg daily) on twenty-nine patients with chronic heart failure in NYHA III-IV stadium who's ejection fraction value was under 40% and had restrictive type of diastolic filling abnormality. Parameters of left ventricular function were measured by echocardiography and the basal data were compared to data detected at the time of 3 and 6 months control. A beneficial effect of enalapril was found on functional cardiac status (reduction in NYHA score), on systolic function (augmentation in ejection fraction and mean mitral anulus motion amplitude). Parallel with the previous parameters in the severity of restrictive diastolic filling alterations (reducing the value of pathologically high E/A ratio and an increasing in deceleration time of early diastolic filling wave) a countable decrease was observed. Authors emphasize the favourable effect of angiotensin converting enzyme inhibitors on chronic heart failure caused by different reasons independently of its severity. The source of drug action is reduction of the preload and afterload as well as keeping under control the harmful ventricular remodelling based on impediment of myocardial angiotensin system.

[Motion of the mitral anulus and suitability of the motion amplitude for determination of left ventricular function in Q-type myocardial infarct]. / A mitralis anulus mozgása és mozgási amplitudójának alkalmassága a bal kamra funkció megítélésére acut Q-típusú myocardialis infarctusban.

Cardiac apex hardly changes its location during the time of cardiac cycle, but the mitral anulus--representing cardiac base--has a systolic descent toward the apex and in diastole moves to the left atrium. The whole amplitude of mitral anulus motion is so useful indicator of left ventricular systolic function as ejection fraction derived from enddiastolic and endsystolic parameters. Mitral anulus motion amplitude easy to measure noninvasively by echocardiography. Authors examined 110 patients with their first, Q-type acute myocardial infarction 7-10 days after the onset of symptoms. In the same time ejection fraction value was assessed by radionuclide ventriculography. Data of ejection fraction and mitral anulus motion amplitude were compared. Their conclusions: 1. mitral anulus motion amplitude well correlated with left ventricular ejection fraction 2. patients had a decreased mitral anulus motion amplitude at the site of infarction 3. investigation of mitral anulus motion amplitude is an useful and easy to make method to assess of systolic function of left ventricle in acute myocardial infarction.

[Acute myocardial infarction mortality in a hospital department during the last decades]. / Az acut myocardialis infarctus halálozásának alakulása osztályunkon az utóbbi évtizedekben.

Authors surveyed the data of 2677 patients with acute myocardial infarction treated in cardiological department between 1970 and 1993. Activity of Coronary Care Unit (has been working from 1974) reduced the earlier hospital mortality in general internal medicine possibilities with 50%. Main cause of this reduction was the temporal electrotherapy of cardiac arrhythmias. Since the middle of last decade the goal of has been generally introduced combined therapy (beta-blockers, intravenous nitrates, acetyl-salicic acid, thrombolytics) is to reduce the mass of damaged myocardium and hereby increasing the ratio of survival. They reduced the hospital mortality around 10% with consistent application of this therapy. According to the authors application of thrombolytic therapy has basic importance in management of myocardial infarction, but they could not treat a lot of patients with streptokinase in consequence of too long prehospital time period and not properly controlled hypertension. The proportion of systemic thrombolytic therapy on their patients is almost a quarter of them. It would be very important to increase this beneficial ratio with reducing the time of patients decision to turn to health care and applying systemic thrombolytic therapy more than six hours after the onset of symptoms of myocardial infarction.

[Unrecognised acute myocardial infarct in a nine-year autopsy material]. / Fel nem ismert acut myocardialis infarctusok elöfordulása 9 év sectiós anyagában.

The authors analysed in a retrospective study the clinical features, ECG and enzyme examinations of 97 patients who died in acute myocardial infarction proved by autopsy during 9 years in their department. They found the recognition rate of the acute myocardial infarction 73%, compared to the data of the literature good. There occurred 26 unrecognized myocardial infarctions, among which atypical clinical features were more frequent and typical ECG signs and enzyme values were less frequent, than among the recognized cases. In their earlier study 57% of the recognized acute myocardial infarctions occurred on the posterior wall, in the present one the rate of anterior and posterior cases was the same. In the earlier study from 95 posterior acute myocardial infarctions 35 (41%) were unrecognized, in present from 41 cases only 11 (27%). This 14% improvement in the recognition rate of the posterior myocardial infarctions should be attributed to the routine application of paravertebral ECG leads, but statistically was not significant.

Clinical trial of ofloxacin alone and in combination with dapsone plus clofazimine for treatment of lepromatous leprosy.

Twenty-four patients with newly diagnosed lepromatous leprosy were allocated randomly to three groups and treated for 56 days with 400 mg of ofloxacin daily, 800 mg of ofloxacin daily, or 400 mg of ofloxacin, 100 mg of dapsone, and 50 mg of clofazimine daily plus 300 mg of clofazimine once every 28 days. The patients in all three groups demonstrated remarkable clinical improvements, accompanied by rapid decline of the morphological index in skin smears during treatment. More than 99, > 99.99, and > 99.99% of the viable Mycobacterium leprae organisms had been killed by 14, 28, and 56 days of treatment, respectively, as measured by inoculation into the footpads of immunocompetent and nude mice of organisms recovered from skin biopsy specimens obtained before and during treatment. Mild to moderate elevations of the serum glutamic pyruvic transaminase level were observed in four patients, all after 28 days of treatment, which returned to normal after the trial had been completed. Clinical improvement, bactericidal activity, and hepatotoxicity did not differ significantly among the three groups. Ofloxacin displayed powerful bactericidal activity against M. leprae in leprosy patients and may be an important component of new multidrug regimens for the treatment of leprosy. Its optimal dosage appears to be 400 mg daily, and combination with dapsone and clofazimine does not enhance its activity.

An attempt to evaluate the risk factors related to coronary heart disease in Hungary.

Using questionnaires, the authors evaluated the risk of coronary heart disease in different groups of Hungarian society. Among physicians, teachers, factory workers and agricultural workers, the latter seem to be at the highest risk. On the basis of these results obtained in a population of 363 agricultural employees, a detailed risk factor analysis was made. These results were compared with those of the Framingham Offspring Study. Hungarians show alarmingly often a high blood cholesterol level, hypertension, smoking and obesity (the latter factor in women). The more frequent occurrence of the three main risk factors (high blood cholesterol, hypertension, smoking) in young Hungarians is concordant with the fact that the incidence of myocardial infarction in young people in Hungary is one of the highest in the world. The frequency of a positive parental history and obesity in men is lower in the Hungarian population than in the American one. The risk of coronary heart disease in the examined Hungarian population is considered high. The authors have launched a preventive programme.

Relationships between PGL-1 antigen in serum, tissue and viability of Mycobacterium leprae as determined by mouse footpad assay in multibacillary patients during short-term clinical trial.

In connection with a 56-day controlled clinical trial for comparing the therapeutic effects between pefloxacin and ofloxacin in 21 lepromatous patients, we have studied the relationships between PGL-1 antigen level in serum and in skin and serum PGL-1 antibody titre on the one hand, and the viability of Mycobacterium leprae, as measured by serial mouse footpad inoculations, and other bactericidal parameters on the other. Before and during treatment, significant correlation was found between serum PGL-1 level and the morphological index (MI), and with the number of viable organisms per mg skin tissue. However, neither serum PGL-1 antibody titre nor skin PGL-1 antigen level showed significant change during the 56-day trial. Because the reduction of serum PGL-1 level was well correlated but less pronounced as compared with the evolution of viable organisms during treatment, the serum PGL-1 antigen assay may be useful as an early indicator of response to chemotherapy in short-term clinical trial, but it is unlikely to replace mouse footpad inoculation for the evaluation of viability of M. leprae.

Clinical trial of pefloxacin and ofloxacin in the treatment of lepromatous leprosy.

Twenty-one previously untreated lepromatous patients were randomized into two groups and treated with either 800 mg pefloxacin (PEFLO) or 400 mg ofloxacin (OFLO) once daily. The trial consisted of two parts: monotherapy from day 0 to day 56; and combined with the World Health Organization multidrug therapy (WHO/MDT) regimen for multibacillary (MB) leprosy from day 57 to day 180. Four patients were removed from the trial because the organisms recovered from their pretreatment biopsies failed to infect mice. Among the remaining 17 cases, four (23.5%) had primary resistance to dapsone but all of them were susceptible to rifampin. The initial (day 0) proportion of viable organisms, as measured by mouse foot pad inoculation, varied tremendously from patient to patient despite randomization during admission. Definite clinical improvement was noticed in virtually all patients after 22 doses of treatment with either PEFLO or OFLO. A significant fall in the morphological index (MI) occurred as early as after 8 doses of PEFLO or after 22 doses of OFLO; the bacterial load also showed a moderate degree of reduction during the period of monotherapy. Although single-dose PEFLO or OFLO displayed only a modest degree of bactericidal effect against Mycobacterium leprae, about 99.9%, or 4 logs, of organisms viable on day 0 were killed by 22 doses of either PEFLO or OFLO. No significant difference in the therapeutic effect was detected between the two regimens. During PEFLO or OFLO monotherapy, except in one patient (case no. 10), the side effects were few and mild. Case no. 10 developed a psychic disorder after 27 days of PEFLO monotherapy, presumably due to the treatment with PEFLO. All of the patients tolerated the period of combined therapy extremely well, although some asymptomatic and transient laboratory abnormalities were observed. Because both PEFLO and OFLO displayed rapid bactericidal activities in human leprosy and were well tolerated by the patients, further clinical trials and field trials in evaluating the therapeutic effects of combined regimens containing both rifampin and PEFLO or OFLO are being organized. Since this is the first clinical trial in leprosy employing nude mice, in combination with normal mice, for monitoring the therapeutic effects of antimicrobials, the advantages, limitations and appropriate timing in using nude mice are discussed.

Effectiveness of pefloxacin in the treatment of lepromatous leprosy.

As a first clinical trial of a fluoroquinolone derivative in leprosy, ten previously untreated lepromatous leprosy patients, about two fifths of them with primary dapsone resistance but all susceptible to rifampin, were treated with pefloxacin 400 mg twice daily for 6 months. Definite clinical improvement was observed in all ten patients as early as 2 months after beginning treatment, and the morphological index was also drastically decreased to the baseline during the same period. The rapid bactericidal effects, as measured by serial mouse foot-pad inoculations, were demonstrated to the extent that about 99% of the bacilli were killed during the first 2 months of treatment. However, the bacterial load, in terms of the bacterial index and the number of acid-fast bacilli per mg of tissue, of the patients was only moderately reduced. The side effects were mild, and the patients tolerated the treatment well.

[Morphologic and biologic properties of diphtherroid organisms (LDC) isolated in leprosy]. / Propriétés morphologiques et biologiques des organismes diphtéroïdes (LCD) isolés chez les lépreux.

Acid-alcohol-fast bacteria are not always detectable in all leprosy lesions. Non acid-fast microorganisms may be associated with acid-fast bacteria. The most frequently isolated strains from leprosy lesions are non acid-fast bacteria, morphologically related to C. diptheirae. Hence their designation as diphtheroïds or LDC (leprosy derived corynebacteria). Their antigenic structure is more closely related to M. leprae and other mycobacteria than to classical corynebacteria. This leads to the hypothesis of a potential role in the pathogenesis of leprosy and their use as an antigen for skin tests by leprosy patients.