Biomedical subjects

P Kucera

Publications and source records attributed to P Kucera.

At least 55 records · Page 3Linked to original sources

PubMed1996

Glucose is arrhythmogenic in the anoxic-reoxygenated embryonic chick heart.

Unlike in adult heart, embryonic myocardium works at low PO2 and depends preferentially on glucose. Therefore, activity of the embryonic heart during anoxia and reoxygenation should be particularly affected by changes in glucose availability. Hearts excised from 4-d-old chick embryos were submitted in vitro to strictly controlled anoxia-reoxygenation transitions at glucose concentrations varying from 0 to 20 mmol/L. Spontaneous and regular heart contractions were detected optically as movements of the ventricle wall and instantaneous heart rate, amplitude of contraction, and velocities of contraction and relaxation were determined. Anoxia induced transient tachycardia and rapidly depressed contractile activity, whereas reoxygenation provoked a temporary and complete cardioplegia (oxygen paradox). In the presence of glucose, atrial rhythm became irregular during anoxia and chaotic-periodic during reoxygenation. The incidence of these arrhythmias depended on duration of anoxia, and no ventricular ectopic beats were observed. Removal of glucose or blockade of glycolysis suppressed arrhythmias. These results show similarities but also differences with respect to the adult heart. Indeed, glucose 1) delayed and anoxic contractile failure, shortened the reoxygenation-induced cardiac arrest, and improved the recovery of contractile activity; 2) attenuated stunning at 20 mmol/L but worsened it at 8 mmol/L; and 3) paradoxically, was arrhythmogenic during anoxia and reoxygenation, especially when present at the physiologic concentration of 8 mmol/L. The last named phenomenon seems to be characteristic of the young embryonic heart, and our findings underscore that fluctuations of glycolytic activity may play a role in the reactivity of the embryonic myocardium to anoxiareoxygenation transitions.

Animals↗

PubMed1996

Endothelin-1 does not mediate the endothelium-dependent hypoxic contractions of small pulmonary arteries in rats.

Various pulmonary artery preparations in vitro demonstrate sustained endothelium-dependent contractions upon hypoxia. To determine whether endothelin-1 could mediate this phenomenon, we examined the effect of bosentan, a new antagonist of both the ETA and ETB subtypes of the endothelin receptor. Small (300 pm) pulmonary arteries from rats were mounted on a myograph, precontracted with prostaglandin F2 alpha and exposed to hypoxia (PO2, 10 to 15 mm Hg, measured on-line) for 45 min. Endothelium-intact control rings exhibited a biphasic response, with a transient initial vasoconstriction (phase 1) followed by a second slowly developing sustained contraction (phase 2). Expressed in percent of the maximal response to 80 mmol/L KCl, the amplitudes of phase 1 (peak tension) and 2 (tension after 45 min of hypoxia) averaged 37 +/- 12% and 17 +/- 14%, respectively (n = 11). In endothelium-denuded rings, phase 1 persisted while the amplitude of phase 2 was reduced to 2 +/- 12% (p < 0.05, n = 8), showing the endothelium dependence of this contraction. Neither phase was significantly decreased in rings treated with 10(-5) mmol/L bosentan (38 +/- 15% and 17 +/- 12%, respectively, n = 6). The PO2 threshold for onset of hypoxic contraction was not significantly different among these three groups and averaged 32 +/- 24 mm Hg. In a separate experiment, we assessed the inhibitory effect of 10(-5) mol/L bosentan on the response to 10(-8) mol/L endothelin-I. Rings treated for 45 min with 10(-8) mol/L endothelin-1 alone exhibited a maximal contraction of 75 +/- 27% (n = 6). This was reduced to 4 +/- 17% (p < 0.01, n = 6) in rings treated with both 10(-8) mol/L endothelin-1 and 10(-5) mol/L bosentan. We conclude that complete blockade of all endothelin receptor subtypes has no effect on either endothelium-dependent or -independent hypoxic contractions in this preparation. This suggests that endothelial factors other than endothelin-I mediate the acute hypoxic contractions of small pulmonary arteries in the rat.

Animals↗

PubMed1996

[Autoimmunity and glutamate decarboxylase antibodies in the pathogenesis of type I diabetes mellitus in experimental and clinical practice].

Diabetes mellitus type 1 (DM type 1) is a chronic, organ specific autoimmune disease. Autoantibodies against the islets of Langerhans (ICA) are found in 71%, against insulin (IA) in 65% against glutamate decarboxylase (GAD) in 67-77% of children with DM type 1; in their close relatives and in patients with a more prolonged persistence of DM type 1 these antibodies and manifestations of cell-mediated autoreactivity are found less frequently also as a sign of the autoimmune character of DM type 1. In the pathogenesis of DM type 1 with the help of experimental mammalian models an idea on the genesis of the disease was obtained--activation of autoimmunity on the background of genetic sensitivity and environmental factors, the development of lymphocytic infiltration of the pancreatic islets (insulitis) destruction of pancreatic B-cells due to the cytotoxicity of T lymphocytes and the manifestation of DM type 1. According to this idea among the series of antigens of B-cells of the islets of Langerhans the dominant antigen is the isoform GAD65--under experimental conditions the specific response of T lymphocytes against GAD65 is found first, then follows extension of the reactivity also against other antigens of the islets. Understanding of the immune pathogenesis of DM type 1 makes it possible to apply the immunomodulating approach to the treatment of this condition which gave the first positive results in experimental work as well as in clinical practice.

Animals↗

PubMed1995

Extracellular protons modulate the Ca2+ block of a Ca(2+)-blockable monovalent cation channel in chick embryo.

Some epithelial nonselective cation channels although in an open conformation are impermeable because they are blocked by extracellular Ca2+ ions. Such a channel is present in the apical membrane of the ectodermal cells of the chick gastrula. This single-channel study shows that protonation of a site located inside the pore of this embryonic channel reduces the Ca2+ block thus allowing Na+ to pass. The apparent pK for Ca2+ at -50 mV shifts from 5.7 at pH 7.4 to 3.5 at pH 4.5. Unexpectedly, the change in Ca2+ affinity does not originate from a destabilization of the binding but comes from a large reduction of the association rate.

Animals↗

PubMed1995

Permeation properties of a Ca(2+)-blockable monovalent cation channel in the ectoderm of the chick embryo: pore size and multioccupancy probed with organic cations and Ca2+.

A Ca(2+)-blockable monovalent cation channel is present in the apical membrane of the ectoderm of the gastrulating chick embryo. We used the patch clamp technique to study several single-channel permeation properties of this channel. In symmetrical conditions without Ca2+, the Na+ current carried by the channel rectifies inwardly. The channel has an apparent dissociation constant for extracellular Na+ of 115 mM at 0 mV and a low density of negative surface charge (-0.03 e/nm2) at its extracellular entrance. The minimal pore diameter is approximately 5.8 A, as calculated from the relative permeabilities of 10 small organic cations. Extracellular application of six large organic cations decreased the inward Na+ current in a voltage-dependent manner, which strongly suggests an intrachannel block. The presence of at least two ion binding sites inside the pore is inferred from the Na+ dependence of the block by the organic cations. This hypothesis is strengthened by the fact that the extracellular Ca2+ block is also modified by the Na+ concentration. In particular, the rise of the unblocking rate with increased Na+ concentrations clearly suggests the presence of an interaction between Ca2+ and Na+ inside the channel. A low probability of double occupancy at physiological ionic conditions is implied from the absence of an anomalous mole fraction effect with mixtures of extracellular Li+ and K+. Finally, the absence of inward current at very strong hyperpolarizations and in the presence of 10 mM extracellular Ca2+ demonstrates the absence of significant Ca2+ current through this channel. It is argued that this embryonic epithelial Ca(2+)-blockable monovalent cation channel is related to both L-type Ca2+ channel and cyclic nucleotide-gated channels.

Animals↗

PubMed1994

Ion transport across the early chick embryo: I. Electrical measurements, ionic fluxes and regional heterogeneity.

The chick blastoderm at the stage of late gastrula is a flat disc formed by three cell layers and exhibiting epithelial properties. Blastoderms were cultured in miniature chambers and their electrophysiological characteristics were determined under Ussing conditions. Under open-circuit condition and identical physiological solutions on both sides, spontaneous transblastodermal potential difference (Voc) of -7.5 +/- 3.3 mV (ventral side positive) was measured. Under short-circuit condition (transblastodermal delta V = 0 mV), the blastoderm generated short-circuit current (Isc) of 21 +/- 8 microA/cm2, which was entirely dependent on extracellular sodium, sensitive to ouabain applied ventrally and independent of extracellular chloride. The net transblastodermal Na+ flux fully accounted for the measured Isc, both under control conditions and with ouabain. The total transblastodermal resistance (Rtot) was 390 +/- 125 omega cm2. Frequently, the Voc, Isc and Rtot showed spontaneous oscillations with a period of 4-5 min. Removal of endoderm and mesoderm did not significantly affect the electrical properties, indicating that the electrogenic sodium transport is generated by the ectoderm. The Voc and Isc measured in the area pellucida (-1.3 +/- 0.8 mV, 9.3 +/- 4.4 microA/cm2) and extraembryonic area opaca (-7.8 +/- 1.1 mV, 31.2 +/- 12.7 microA/cm2) were significantly different. Such a heterogeneous distribution of electrical properties can explain the presence in the blastoderm of extracellular electrical currents found by using a vibrating probe.

Animals↗

PubMed1994

Ion transport across the early chick embryo: II. Characterization and pH sensitivity of the transembryonic short-circuit current.

The ectoderm of the one-day chick embryo generates dorsoventrally oriented short-circuit current (Isc) entirely dependent on extracellular sodium. At the dorsal cell membrane, the Isc was modified reversibly and in a concentration-dependent manner by: amiloride (60% decrease at 1 mM, with 2 apparent IC50S: 0.13 and 48 microM), phlorizin (0.1 mM) or removal of glucose (30% decrease, additive to that of amiloride), SITS (1 mM, 13% decrease). Acidification of alkalinization of the dorsal (but not ventral) superfusate produced, respectively, decrease or increase of Isc with a pH50 of 7.64. Ba2+ (0.1-1 mM) from either side of the ectoderm decreased the Isc by 30%. Anthracene-9-carboxylic acid, furosemide and inducers of cAMP had no effect on electrophysiological properties of the blastoderm. The chick ectoderm is therefore a highly polarized epithelium containing, at the dorsal membrane, the high and low affinity amiloride-sensitive Na+ channels, Na(+)-glucose cotransporter, K+ channels and pH sensitivity, and, at the ventral membrane, the Na+, K(+)-ATPase and K+ channels. The Na+ transport reacts to pH, but lacks the cAMP regulatory system, well known in many epithelia. The active Na+ transport drives glucose and fluid into the intraembryonic space, across and around the blastoderm which, in the absence of blood circulation, could secure renewal of extracellular fluid and disposal of wastes and thus maintain the cell homeostasis.

Amiloride↗

PubMed1994

Cation channel blocked by extracellular Ca2+ in the apical membrane of the chick embryonic ectoderm.

In the chick embryo (20 h incubation, gastrula stage), the apical membrane of the ectodermal cells shows a high density of a non-selective cation channel which is blocked by very low extracellular Ca2+ concentrations. Properties of this channel were studied at the single-channel level using the patch-clamp technique in the cell-attached mode. With 1 mmol/l Ca2+ in the pipette, only outward current was present and the channel conductance measured at +120 mV was 25.5 pS. In the absence of Ca2+, also inward current through the channel was observed. The conductances measured at -50 mV were 49.5 pS with Na+ as the charge carrier, 72.5 pS with K+, 49.1 pS with Cs+, and 18.5 pS with Li+. The conductance measured at +80 mV was around 23 pS in all four cases. The reversal potential was similar (around 25 mV) for all four ions, which indicates a poor selectivity of the channel. In the absence of Ca2+ and the presence of 1 mmol/l ethylenebis(oxonitrilo)tetraacetate (EGTA), the kinetics of the channel were characterized by bursts of the order of seconds. During a burst, the channel flickered between one open and one closed level. The open time was constant between -30 mV and -80 mV, while the closed time decreased with hyperpolarization. The open time varied according to the permeant ion (K+ < Na+ = Cs+ < Li+). Extracellular Ca2+ blocked the inward current in a voltage-dependent manner. The Kd values, 1 mumol/l at -30 mV and 3.2 mumol/l at -80 mV, indicate that Ca2+ ions exit the channel toward the intracellular side. A weak voltage dependency of the association rate constant suggests that the Ca(2+)-binding site is close to the outside mouth. Extracellular Ca2+ was much less efficient at blocking the outward current (Kd about 1 mmol/l at 80 mV). Tetracaine, but not uraniumdioxide, decreased the opening probability of the channel. The embryonic channel shows similarities with the Ca(2+)-blockable, poorly selective channel described in the epithelium of toad urinary bladder.

Animals↗

PubMed1994

Origins of motility patterns in isolated arterially perfused rat intestine.

BACKGROUND/AIMS: Assessment of the neuromuscular control of small intestinal motility and movement of luminal contents is hampered in vivo by measurement techniques and in vitro by tissue viability. The aim of this study was to establish the structural and functional integrity of an isolated segment of rat ileum and characterize its motility. METHODS: Segments of rat ileum were perfused arterially with oxygenated fluorocarbon and luminally with saline. Oral and aboral pressures were correlated with conformational changes detected by concurrent video imaging. RESULTS: Light and electron microscopy showed no neuromuscular abnormalities after experiments, and acetylcholine-induced pressure amplitudes were unchanged during experiments. Under basal conditions, low-frequency contractions showing constant frequency (0.27/min) and amplitudes (oral, 17 hPa; aboral, 15 hPa) corresponded to luminally occlusive aborally propagated contractions, which were eliminated by tetrodotoxin. High-frequency contractions with a constant frequency (27/min) were also seen; their basal amplitude (0.3 hPa) increased immediately before and after low-frequency contractions and after tetrodotoxin. Tetrodotoxin also increased basal intestinal tone. CONCLUSIONS: An isolated, arterially perfused segment of rat ileum retains structural and functional integrity. It shows low-frequency propulsive contractions, controlled by the enteric nervous system, and myogenic high-frequency contractions, probably subject to tonic neural inhibition.

Acetylcholine↗

PubMed1993

Embryonic and fetal development: fundamental research.

Much progress has been made over the past decades in the development of in vitro techniques for the assessment of chemically induced effects in embryonic and fetal development. In vitro assays have originally been developed to provide information on the mechanism of action of normal development, and have hence more adequately been used in fundamental research. These assays had to undergo extensive modification to be used in developmental toxicity testing. The present paper focuses on the rat whole embryo culture system, but also reviews modifications that were undertaken for the in vitro chick embryo system and the aggregate cultures of fetal rat brain cells. Today these tests cannot replace the existing in vivo developmental toxicity tests. They can, however, be used to screen chemicals for further development or further testing. In addition, these in vitro tests provide valuable information on the mechanisms of developmental toxicity and help to understand the relevancy of findings for humans. In vitro systems, combined with selected in vivo testing and pharmacokinetic investigations in animals and humans, can thus provide essential information for human risk assessment.

Animals↗

PubMed1992

Oxygen uptake during early cardiogenesis of the chick.

Oxidative metabolism of the isolated embryonic heart of the chick has been determined using a spectrophotometric technique allowing global as well as localized micromeasurements of the O2 uptake. Entire hearts, excised from embryos of 10 somites (primordia fused, stage 10 HH) and 40 somites (S shaped, stage 20 HH) were placed in a special chamber under controlled metabolic conditions where they continued to beat spontaneously and regularly. During the 32 h of development, the O2 consumption of the whole heart increased from 0.9 +/- 0.1 to 5.3 +/- 0.8 nmol O2/h. These values corrected for protein content were, however, comparable (0.45 nmol O2.h-1.micrograms-1). At stage 10-12, the O2 uptake varied along the cardiac tube (from 0.74 to 1.0 nmol O2.h-1.mm-2). From stage 10 to 20, the O2 uptake per unit area of ventricle wall increased from 0.7 +/- 0.2 to 1.8 +/- 0.2 nmol O2.h-1.mm-2, and the O2 uptake per myocardial volume during one cardiac cycle varied from 7 to 2.5 nmol O2/cm3. These results indicate that, despite an intense morphogenesis, the cardiac tissue has a rather low and stable oxidative metabolism, although the O2 requirement of the whole heart increases significantly. Moreover, the normalized suprabasal aerobic energy expenditure decreases throughout early cardiogenesis. The functional integrity of the isolated embryonic heart combined with the experimental possibilities of the microtechnique make the preparation appropriate for studying the changes in cardiac metabolism during development.

Animals↗

PubMed1991

Fibronectin distribution in the chick embryo during formation of the blastula.

The distribution of the fibronectin-rich extracellular matrix (ECM) in the chick embryo during formation of the blastula has been evaluated semiquantitatively using an electron microscopical immunogold staining technique. During the first 10 h of postlaying development, fibronectin was found in both embryonic area pellucida and extra-embryonic area opaca of the blastoderm. In the area pellucida, the fibronectin was (1) associated with the basal lamina of the epiblast, (2) present between epiblastic and hypoblastic cells and (3) occasionally internalized in hypoblastic cells. Along the embryonic axis, a transient and high density of ECM was associated with the front of the anteriorly and rapidly expanding hypoblast. Very high density of fibronectin was observed in the marginal zone of the area pellucida, where the epiblastic and deeper cell layers show contacts and intense re-arrangements. In the area opaca, fibronectin was at first found only sporadically between contacting cells, but its density increased steadily and markedly during the first day of development. These rapid and significant changes in the regional distribution of fibronectin-rich ECM are discussed with respect to the early morphogenesis of the chick embryo.

Animals↗

PubMed1991

Oxygen uptake of isolated toad skin epithelium: micromeasurement and effect of ionic acclimation.

Oxidative metabolism of isolated toad skin epithelium (Bufo viridis) was investigated in vitro under open-circuit conditions using the spectrophotometric oxyhemoglobin micromethod. This highly sensitive technique has been adapted for studying several epithelia in parallel and for detecting possible regional variations of oxygen uptake in individual epithelium. Changes in the proportion of mitochondria-rich cells (MRC) by ionic acclimation affected oxidative metabolism under nontransporting condition. After acclimation of animals to either NaNO3 or NaCl solutions (100 mmol/l, for greater than 2 wk), the number of MRC per square millimeter in epithelia from nonacclimated and NaNO3- and NaCl-acclimated animals was 350 +/- 113, 460 +/- 196, and 107 +/- 52, respectively. O2 uptake of nonacclimated and NaNO3-acclimated epithelia was significantly higher than that of NaCl-acclimated epithelia (i.e., 0.89 and 0.90 vs. 0.57 nmol O2.h-1.mm-2, respectively). The correlation established between O2 uptake and number of MRC allowed evaluation of the respiration rate of one single MRC, i.e., approximately 1 pmol O2/h. The lowest mitochondrial oxidative activity was found in the epithelia from NaCl-acclimated toads where the uncoupler 2,4-dinitrophenol (50 mumols/l) had the highest relative stimulatory effect (+114%). Acetazolamide (50 mumols/l), a potent inhibitor of carbonic anhydrase mainly present in the MRC, reduced selectively by 31% O2 uptake of the MRC-rich epithelia (NaNO3 acclimated). O2 uptake increased significantly by approximately 80% when basolateral pH increased from 5.8 to 7.8, but did not depend on apical pH. These findings indicate that under nontransporting (open-circuit) conditions, aerobic metabolism of the isolated toad skin epithelium is related to the density and/or characteristics of the MRC.(ABSTRACT TRUNCATED AT 250 WORDS)

Acclimatization↗

PubMed1990

[Surgical treatment of Zenker's diverticulum].

The authors present an account of diagnostic methods, endoscopic problems and surgical treatment of Zenker's diverticula of the oesophagus. They analyze possibilities of X-ray and endoscopic diagnosis; a frequent problem is the insertion of an oesophageal probe before operation; tamponade of the diverticula before operation is useful. Surgical treatment is indicated in major diverticula, small flat ones are followed up. The surgical approach is from the left side of the neck. In the authors' department always total resection of the dicerticulum was performed in major diverticule, invagination could be used in minor diverticula. Early and late complications of operations are discussed.

Aged↗

PubMed1990

An experimental model for measuring middle ear antimicrobial drug penetration in otitis media.

Bacteria are an important cause of acute otitis media and successful treatment depends on achieving inhibitory or bacteriacidal antimicrobial drug concentrations in the middle ear. To evaluate further otitis media treatment success and failure, we developed a chinchilla model to study antimicrobial drug penetration through the middle ear mucosa. Using quantitative histomorphometry, we measured the middle ear space in 10 chinchillas and found a mean +/- SD volume of 2.09 +/- 0.08 ml and a mean +/- SD surface area of 14.41 +/- 1.48 cm2. To measure the apparent rate constant (Kc) of antibiotic elimination from the middle ear, through the middle ear mucosa, an antibiotic solution was inoculated into the middle ear cavity, and samples were aspirated between 1 and 8 hr later. In normal ears, the mean Kc +/- SD for amoxicillin was 0.118 +/- 0.013 hr-1, that for a trimethoprim 0.461 +/- 0.090 hr-1, and that for sulfamethoxazole 0.265 +/- 0.062 hr-1. In ears inoculated with type 7F Streptococcus pneumoniae to induce acute otitis media, the Kc +/- SD increased for all three drugs (P less than 0.05): amoxicilin to 0.286 +/- 0.089 hr-1, trimethoprim to 0.662 +/- 0.118 hr-1, and sulfamethoxazole to 0.411 +/- 0.056 hr-1. These values demonstrate that amoxicillin had the lowest apparent penetration rate constant of the three antibiotics but the greatest increase from normal to infected mucosa (142%). Trimethoprim had the highest apparent penetration rate constant of the three antibiotics but the smallest increase from normal to infected mucosa (44%), while the sulfamethoxazone apparent penetration rate constant increased from normal to infected mucosa by 55%.(ABSTRACT TRUNCATED AT 250 WORDS)

Amoxicillin↗

PubMed1990

[Reconstruction of the facial skeleton using Pallacos R].

Defects of the facial skeleton cause frequently severe aesthetic facial changes with negative repercussions also on the patient's mental state. The skeleton may be affected by injury, a neoplastic disease and iatrogenically when it is necessary to remove a part of the skeleton during operation. One possible way how to replace and reconstruct the skeleton is to use bone cement Pallacos R. This method was used in 8 patients with skeletal affections. The use of Pallacos R has the advantage that it can be used quickly, that it is easy to work with directly at the site of the defect and that its tolerance is satisfactory. It permits also to preserve the physiological shape of the paranasal sinuses. The method is one approach to individually indicated reconstruction.

Bone Cements↗

PubMed1990

[Treatment of tinnitus with gentamicin].

Tinnitus is a very serious problem in otolaryngology. The authors used for torpid cases the intratympanic administration of Gentamicin sulphate, administered twice a day. Nine patients with Ménière's disease were treated. The results were very favourable, deterioration of hearing did not occur and the tinnitus disappeared or improved markedly.

Female↗

PubMed1989

[Rational treatment of necrotizing external otitis].

Advances in the diagnosis and treatment of necrotizing external otitis (NEO) during the past 20 years improved substantially the prognosis of this disease. The use of hyperbaric oxygen therapy greatly potentiates the effect of newly developed highly effective antibiotics. Radioisotope examination of the petrous bones facilitates the diagnosis and indicates the time to terminate treatment. Medicamentous treatment is preferable to surgical treatment, which is reserved for exceptional very advanced cases of the disease. As NEO has been known for 30 years and the means to control it have been improved, it is no longer "malignant" for the majority of patients where the disease is diagnosed in time and treated adequately.

Aged↗