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Transient receptor potential cation channel subfamily M member 4 (TRPM4) is a Ca2+-activated nonselective cation channel that mediates membrane depolarization. Although, a current with the hallmarks of a TRPM4-mediated current has been previously reported in pancreatic acinar cells (PACs), the role of TRPM4 in the regulation of acinar cell function has not yet been explored. In the present study, we identify this TRPM4 current and describe its role in context of Ca2+ signaling of PACs using pharmacological tools and TRPM4-deficient mice. We found a significant Ca2+-activated cation current in PACs that was sensitive to the TRPM4 inhibitors 9-phenanthrol and 4-chloro-2-[[2-(2-chlorophenoxy)acetyl]amino]benzoic acid (CBA). We demonstrated that the CBA-sensitive current was responsible for a Ca2+-dependent depolarization of PACs from a resting membrane potential of −44.4 ± 2.9 to −27.7 ± 3 mV. Furthermore, we showed that Ca2+ influx was higher in the TRPM4 KO- and CBA-treated PACs than in control cells. As hormone-induced repetitive Ca2+ transients partially rely on Ca2+ influx in PACs, the role of TRPM4 was also assessed on Ca2+ oscillations elicited by physiologically relevant concentrations of the cholecystokinin analog cerulein. These data show that the amplitude of Ca2+ signals was significantly higher in TRPM4 KO than in control PACs. Our results suggest that PACs are depolarized by TRPM4 currents to an extent that results in a significant reduction of the inward driving force for Ca2+. In conclusion, TRPM4 links intracellular Ca2+ signaling to membrane potential as a negative feedback regulator of Ca2+ entry in PACs.

Over the past 30 years, transient receptor potential (TRP) channels have evolved from a somewhat obscure observation on how fruit flies detect light to become the center of drug discovery efforts, triggering a heated debate about their potential as targets for therapeutic applications in humans. In this review, we describe our current understanding of the diverse mechanism of action of TRP channels in the itch pathway from the skin to the brain with focus on the peripheral detection of stimuli that elicit the desire to scratch and spinal itch processing and sensitization. We predict that the compelling basic research findings on TRP channels and pruritus will be translated into the development of novel, clinically useful itch medications.

Clinical findings indicate that Strogen forte (a standardized extract of the plant Sabalis serrulata) can be use successfully in the medical treatment of benign prostatic hyperplasia. The aim of the present study was to investigate the possible inhibitory effects of the Strogen forte extract on rat and human testicular delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSD) and prostatic 5 alpha-reductase (5 alpha-R). Strogen forte proved to be a direct inhibitor of medium effectivity, with similar IC50 values for rat testicular delta 5-3 beta-HSD (400 +/- 23 micrograms/ml) and human testicular delta 5-3 beta-HSD (212 +/- 8.6 micrograms/ml). 5 alpha-R activities were analysed by in vitro incubation of rat and human prostatic tissue homogenates with 14C-labelled testosterone as substrate in KRPG-DTT medium (pH = 7.4) with NADPH coenzyme, in air, at 37 degrees C, in the presence of Strogen forte extract. The results clearly demonstrate that Strogen forte is a potent inhibitor of prostatic 5 alpha-R, with IC50 values of 385 +/- 35.6 micrograms/ml for rat and 245 +/- 64.6 micrograms/ml for human prostatic 5 alpha-R. The present study has revealed that this plant extract inhibits not only prostatic 5 alpha-R, but also testicular delta 5-3 beta-HSD.

Both human investigations and studies in animal models have suggested that abnormalities in GABA(A) receptor function have a potential role in the pathophysiology of absence seizures. Recently we showed that, prior to seizure onset, GABA(A) IPSCs in thalamic reticular (NRT) neurons of genetic absence epilepsy rats from Strasbourg (GAERS) had a 25% larger amplitude, a 40% faster decay and a 45% smaller paired-pulse depression than those of nonepileptic control (NEC) rats. By means of a novel mathematical description, the properties of both GAERS and NEC GABAergic synapses can be mimicked. These model synapses were then used in an NRT network model in order to investigate their potential impact on the neuronal firing patterns. Compared to NEC, GAERS NRT neurons show an overall increase in excitability and a higher frequency and regularity of firing in response to periodic input signals. Moreover, in response to randomly distributed stimuli, the GAERS but not the NEC model produces resonance between 7 and 9 Hz, the frequency range of spike-wave discharges in GAERS. The implications of these results for the epileptogenesis of absence seizures are discussed.

All three forms of recombinant low voltage-activated T-type Ca(2)(+) channels (Ca(v)3.1, Ca(v)3.2 and Ca(v)3.3) exhibit a small, though clearly evident, window T-type Ca(2)(+) current (I(Twindow)) which is also present in native channels from different neuronal types. In thalamocortical (TC) and nucleus reticularis thalami (NRT) neurones, and possibly in neocortical cells, an I(Twindow)-mediated bistability is the key cellular mechanism underlying the expression of the slow (1 Hz) sleep oscillation, one of the fundamental EEG rhythms of non-REM sleep. As the I(Twindow)-mediated bistability may also represent one of the cellular mechanisms underlying the expression of high frequency burst firing in awake conditions, I(Twindow) is of critical importance in neuronal population dynamics associated with different behavioural states.

We examined on models the inner pressure tolerance of the most frequently performed single layer anastomoses used in esophagectomies and gastrectomies. The aim was to examine whether interrupted or continuous suture proves safer immediately after the operation. We investigated the difference between sutures involving and not involving the mucosa. The anastomosis models were of organs of hybrid pigs slaughtered in the meet-industry. Atraumatic, 3/0 Biosyn suture material was used. The bursting pressure was measured by insufflating CO2 gas. Its bursting pressure can characterize the inner pressure tolerance of an anastomosis. The circumstances of the experiment, the suture technique and the examination of the bursting pressure were standardized. Our considerations are: 1. The early inner pressure tolerance of an anastomosis does not depend on the condition of the wall. The critical factor is their ability to expand. 2. The inner pressure tolerance of continuous sutures is better than of interrupted ones if the tissues in the suture line show similar degree of dilatation. Optimal conditions are achieved with end-to-end or side-to-side anastomoses of single organs (for example between small bowel and small bowel or colon and colon). 3. Involving the mucosa in the stitches has not influenced early physical suture certainty.

Echovirus 11' (prime) isolates from an epidemic of haemorrhagic syndrome in departments of obstetrics in Hungary have been characterised. The leading component of the clinical disease was carditis and its lethal outcome occurred in 13 newborn babies. Maternal immunity was found to be absent even in women of 41 years of age. The application of monovalent oral poliovirus type 1 vaccine prevented the progress of the epidemic within two weeks. Nevertheless, a serological survey among primovacinees of 3-15 months of age revealed that 20% of the babies seroconverted without clinical symptoms during the epidemic. Serological evidence showed that the echovirus 11' infection was unable to interfere with the efficacy of oral poliovirus vaccine (OPV), since seroconversion rates of primovaccinees did not differ significantly from those in the group seroconverted also to echovirus 11' during the vaccination campaign. A 440 nucleotide (nt) fragment of the 5'-non-translated region of 12 epidemic echovirus 11' isolates and 26 echovirus prototype strains was amplified by a nested reverse transcription-polymerase chain reaction (RT-PCR) and analysed using three different restriction endonucleases. The 5'-regions of the echovirus 11' isolates were found to be identical to each other but different from that of the prototype echovirus 11 (Gregory) strain. The results indicate that echovirus 11' isolates underwent genetic changes in the 5'-end and P1 region of the genome before the onset of the epidemic.

Diagnostic value of multiplex polymerase chain reaction (PCR) was examined by using three primer pairs, specific for the common conserved region of stx1 and stx2, eae and an enterohaemolysin A gene (ehxA). The sensitivity in respect of each amplicon decreased with three exponents comparing to the individual PCR reactions. These PCR reactions were partially inhibited by the presence of certain additional primers. This inhibitory effect was template-concentration dependent, and was partially balanced by usage of increased amount of dNTP. Taq DNA polymerase in a range of 0.3-1.25 U/reaction did not influence the inhibition. The same inhibition was detected if the annealing temperature was changed from 48 degrees C to 57 degrees C. Pairs of EHEC primers inhibited a Salmonella enteritidis virulence-plasmid specific gene amplification, as well. Theoretical inhibiting effects were predicted by Primer Premier software but our observations can be sufficiently explained neither by the competitions between the specific and aspecific amplifications nor by the inhibition caused by dimerization of primers.

The binding of androgens and structurally related analogues to the androgen receptor was studied. The in vitro experiments were carried out with cytosol of castrated rat prostate, using [3H]R1881 (methyltrienolone) as radioligand. The binding parameters measured were Kd = 1.25 x 10(-10) M and Bmax = 111 fmol (mg protein)-1. Ligand specificity was confirmed by competition experiments with known androgen, oestrogen and progestogen ligands. The receptor binding of substituted steroids was studied. The RBAs (relative binding affinities) of our recently synthetized 16-alkyl steroids were low. The only exception was the 17 beta-hydroxy-16 beta-methylestr-4-en-3-one, which exhibited the remarkable RBA of 22.9%.

Specific and sensitive radioimmune methods were used to study the steroid levels and binding capacities of their binding proteins in the serum of 30 women with androgenetic alopecia, in order to establish the proportions of the patients in whom hyperandrogenism or hypercorticism can be detected. It was found that the serum testosterone, dehydroepiandrosterone sulphate and cortisol levels did not differ from those for healthy women except dehydroepiandrosterone which was high. In 5 cases (16.6%), the binding capacity of the "sex hormone-binding globulin" was pathologically low, while in 6 patients (20%) the "free androgen index" was elevated. The binding capacity of the "corticosteroide-binding globulin" was pathologically low in 4 of 21 cases (19%) while the "free cortisol index" was high in 5 of these 21 patients (23.8%). Diane treatment (2 mg cyproterone acetate and 50 micrograms ethinyl-oestradiol)--was administered during 2-5 cycles (8 cases), the pathological dehydroepiandrosterone and dehydroepiandrosterone sulphate levels normalized, the testosterone level decreased to the lower limit of the normal range, the binding capacity of the "sex hormone-binding globulin" increased considerably, and the "free androgen index" fell below the normal range. Diane treatment, an essential improvement was observed in the condition of the patients: the hair became less greasy and its tendency to fall out was likewise moderated.

A total of 93 wild type Escherichia coli of human origin (mostly representing intestinal isolates from Hungary) were examined for the presence of Shiga-like-toxin (SLT) genes using SLT-I, SLT-II and enterohemorrhagic E. coli (EHEC) specific DNA probes. The structural genes of the above specificity were labelled by random priming using 32-P-dCTP. E. coli strains investigated represented 16 serogroups: O1, O2, O4, O5, O6, O18, O25, O26, O45, O55, O111, O125, O126, O128, O157, O165, members of which were likely to produce verotoxin (VT) and strains of serotypes O11:NM, ONT:NM isolated from six haemorrhagic uraemic syndrome (HUS) patients. Out of these strains 51 were examined for in vitro VT production capacity. Only one strain (an O26:H11 from Germany) produced VT. This was also the only strain which proved to be positive with one of the SLT probes (SLT-II), and with the EHEC probe. From this strain a phage was isolated which was proven to be nonconvertible. These data support epidemiological and clinical observations about the very low occurrence or absence of EHEC in Hungary, in contrast to many European countries.

A simple and rapid method of measuring 5 alpha-reductase (5 alpha-R) activity and of determining the kinetic parameters (KM and Vmax) of the enzyme is described. The 5 alpha-R activity in the homogenate of the prostate of Wistar rats aged 8-12 weeks was established, and the effects of natural and synthetic steroids and of non-steroidal antiandrogens (IC50) upon the 5 alpha-R activity were studied. Of the natural steroids, 17-OH-progesterone was found to have the highest inhibitory effect (IC50 = 1.35 microM), followed in decreasing order by progesterone (IC50 = 5.0 microM) and 4-androstene-3,17-dione (IC50 = 21.6 microM). Oestradiol-17 beta had practically no inhibitory effect. Of the synthetic steroids, 4-MA had the highest inhibitory effect (IC50 = 0.068 microM), followed by nortestosterone (IC50 = 7.4 microM) and RU-486 (Mifepristone) (IC50 = 115 microM). Even at 1000 microM, cyproterone acetate exerted no inhibitory effect. Of the nonsteroidal compounds, ketoconazole proved a weak inhibitor (IC50 = 115 microM), while flutamide was practically ineffective.

Models of ion channel kinetics have previously assumed that the switching between the open and closed states is an intrinsically random process. Here, we present an alternative model based on a deterministic process. This model is a piecewise linear iterated map. We calculate the dwell time distributions, autocorrelation function, and power spectrum of this map. We also explore non-linear generalizations of this map. The chaotic nature of our model implies that its long-term behavior mimics the stochastic properties of a random process. In particular, the linear map produces an exponential probability distribution of dwell times in the open and closed states, the same as that produced by the two-state, closed in equilibrium open, Markov model. We show how deterministic and random models can be distinguished by their different phase space portraits. A test of some experimental data seems to favor the deterministic model, but further experimental evidence is needed for an unequivocal decision.

On a large, closed pig farm using artificial insemination (AI), 29 sows were treated with Prolan S-öl injection (Bayer, FRG) and 31 sows with a GnRH superactive analog (Ovurelin, D-Phe6-GnRH-EA, Reanal, Hungary) 48 h after weaning. The effect of treatment on the sows' sexual function was monitored by serum progesterone radioimmunoassay (RIA). The conception rate in the control group (36 sows) was 69.4%. In the groups treated with Prolan and Ovurelin it was 79.3 and 71%, respectively. The use of Prolan S-öl injection markedly reduced the number of acyclic sows and of those having an irregular oestrous cycle. Treatment with the GnRH analog inhibited the manifestation of weaning-induced heat; subsequently, however, it induced a regular cycle in 30 out of the 31 sows treated.

The consequences of preservation by solution of extracellular composition were examined in 24 renal autografts. Nine animals survived for indefinite period. In the reimplanted kidneys of the animals which had died in the acute phase (1st to 7th day) gross oedema, haemorrhages, lesions of tubular epithelium and glomerular congestion were prevalent. In later stages (23rd to 43rd day) the renal structure was well preserved in some places and there were extensive fibrous peritubular areas. In the kidneys of the long-term survivors the fibrous tissue resulting from irreversible destruction was confined to small areas. It is concluded that, though some degree of tissue destruction is inevitable, it is possible to minimize it by using appropriate preserving solutions.

Using the concept of the Markov dependency, we analyzed the spontaneous activity in the pre- and postganglionic nerve trunks of the feline superior cervical ganglion. The time structure (order and value of time dependency) of the sequence of compound action potentials was determined. The analysis revealed that the time dependency of the postganglionic activity is of higher order and is generally more consistent than that of the preganglionic one. The difference in orders and values between the pre- and postganglionic activities was found to be a measure of the integrative function of the ganglion. A transformation by the ganglion of some additional information, inaccessible in the time structure of the preganglionic activity, seems to underlie this difference.

The fine structural characteristics of ZIO reaction was studied in the cerebral and cerebellar cortex and olfactory bulb of the rat and in synaptosomes prepared from rat cerebellar cortex. It was concluded that: 1. Organelles of different nerve cell types exhibit different ZIO reactions provided that the impregnation was carried out under standardized conditions. 2. 6...10 times more synaptic vesicles were stained by ZIO in the inhibitory terminals than in the excitatory ones. 3. ZIO positivity was found in all types of synaptosomes prepared from cerebral cortex. Following electrical or chemical (KCl) depolarization there was a decrease in the number of ZIO positive synaptic vesicles, which decrease was directly proportional to the parameters of stimulation. 4. By x-ray microanalysis Os, Zn and Ca were consistently detected in the ZIO precipitates. Iodine, however, could not always be found. After stimulation the presence of Ca was observed even in those synaptosomes in which the ZIO reaction product was absent. 5. On the basis of the staining characteristics the reaction, under standard conditions, can reflect certain functional states of the nerve terminals.

Four different preserving fluids were studied for protective effect in a total of 33 isolated canine kidneys. The preservation method consisted in initial perfusion followed by 24 hours of hypothermic storage. Two electrolyte and two non-electrolyte solutions were tested. One of the electrolyte solutions was of the extracellular, the other of the intracellular type. The two non-electrolyte solutions were 5% human albumin and 10% Rheomacrodex.

Donor kidneys were preserved in a Collins-4 solution containing periodate in concentrations of 10(-2) and 10(-4) mole/l, respectively, in an attempt to oxidize the carbohydrate components of the antigen-carrying membrane surfaces and thus to influence the rejection phenomena exhibited by the allogeneic kidney. All kidneys were, however, rejected as a result of a hyperacute reaction indicating that periodate did not react with the hydrophobic membrane structures in vivo. Not only did the antigen thus remain unchanged but the vulnerability of other renal structures to the destructive effect of the killer cells also increased as a result of toxic damage.

The possibility of structural and functional preservation of the isolated liver has been studied in dog experiments. A sterile liver preparation left in situ was worked out, having practically no ischaemic period, except during perfusion. Nevertheless, well-defined lesions (perivascular and interstitial oedema, dissociated contraction of the smooth muscle elements of the veins, cellula accumulation in the sinusoids) appeared, accompanied by a gradual reduction (microcirculatory disorders, enzyme derailment) of the function of hepatocytes.

In the present experiments the role of unmyelinated sensory fibres in the mechanism of cutaneous inflammatory reactions under normal and pathological conditions has been studied in man and animals. Dye leakage responses to histamine, serotonin, compound 48/80, bradykinin and substance P were significantly reduced, while neurogenic inflammation was completely abolished in rats treated neonatally with capsaicin, as studied quantitatively by the Evans blue technique. Neurogenic inflammation could also be elicited by mustard oil in normally innervated human skin, but not in skin areas affected by herpes zoster or in a patient suffering from congenital analgesia. Repeated topical treatment of the skin with capsaicin (local desensitization) abolished the neurogenic inflammatory response for several days. Chemical pain sensitivity was strongly reduced, and thresholds for warmth and heat pain sensations were significantly elevated. Local capsaicin desensitization of the skin prevented whealing, flare and itch in patients with acquired cold and heat urticaria. The findings indicate that peptide-containing sensory nerves are involved in the mediation of chemogenic and heat pain, and possibly itch, and are responsible for initiation of the neurogenic inflammatory response. The results also provide direct evidence of the involvement of these particular sensory nerves in the modulation of the permeability-increasing effects of putative mediators of acute inflammatory reactions. It is concluded that, through modulation of cutaneous vascular reactions, peptidergic sensory nerves may play a hitherto unrecognized role in the pathomechanism of certain diseases of human skin.

Various observations suggest that sympathetic ganglia act as local integrative centers redistributing preganglionic excitation (i.e. the information issued by the central nervous system) to the postganglionic fibers (and effector organs). In order to support this concept a simple mathematical model of the elementary integration process, treating the case of a single preganglionic compound action potential, has been developed. This quantitative description, based on a few elementary assumptions, shows a possible way of processing preganglionic excitation in the ganglion. It is shown that on a particular ganglion cell the probability distribution of the number of activated synapses obeys hypergeometric distribution and hence, the postganglionic compound action potential is built up of several compound action potentials occurring at different times. The former correspond to different groups of firing cells. The model discloses modes of structural and temporal pattern generation performed by the sympathetic ganglion.

The structure of the dog kidney can be preserved in hypothermic intracellular-type solutions, but after 6 hours the appearance of pathologic focal changes such as mild oedema, lesions of the cellular elements, podocyte heteromorphism, vacuolization, disintegration of the basal membrane and of the cytomembrane structrues and even cell degeneration, is unavoidable.

Shock was elicited in anaesthetized dogs by maintaining a haemorrhagic hypotension of 4 kPa until 30 per cent spontaneous refusion, followed by total reinfusion. Functional residual capacity (FRC) and minute ventilation increased considerably similarly to our previous experiments. Lactate content in both the external intercostal and the biceps femoris muscles increased significantly in advanced shock. The expiratory external abdominal oblique muscle showed electromyographic signs of fatigue. At the height of the FRC changes tonic contraction of the external intercostal muscle could be demonstrated electron microscopically. This tonic contraction is the main factor in the large FRC rise in late shock forming the basis of a hitherto unknown vicious circle.

Animal experiments were carried out on 12 dogs. Dogs are specially qualified because the choledochus and the pancreatic duct seperately fall into the duodenum. Laboratory tests demonstrated the influence of the choledocholigature on the exocrine function of the pancreas. These findings refer to the unity of the biliary tract and the pancreas. The histologic inquiries of pancreas did not showed any pathologic alterations because the survival time was too short likely.

The reproductive status of sows was monitored by serum progesterone radioimmunoassay (RIA) in a closed pig herd of 1100 sows, producing for the market. The method proved suitable for monitoring reproductive events (progesterone production during lactation, effect of weaning on the resumption of cycling, successful and unsuccessful inseminations, abortion, etc.) and for diagnosing ovarian disturbances (acyclia, irregular cycle, etc.). A total of 813 inseminations were followed up to determine the time of return to oestrus. The serum progesterone values of blood samples taken from the v. cava cranialis on day 22 after insemination indicated that pregnancy was diagnosed with a reliability of 75.5%, while non-pregnant sows failing to return to oestrus were identified with a reliability of 96.1%. The economic benefits to be derived from the use of this method in a pig herd are discussed.