Reduction of myocardial ischemia by gallopamil: a dual-isotope study with thallium-201 and iodine-123 phenylpentadecanoic acid.

The present study was performed for characterizing the effect of chronic oral treatment with the calcium antagonist gallopamil on regional myocardial perfusion and free fatty acid utilization in poststenotic human myocardium. Twenty-two patients with angiographically documented coronary artery disease and stable angina pectoris underwent consecutive dual-isotope studies following simultaneous injection of 80 MBq thallium-201 and 200 MBq iodine-123 phenylpentadecanoic acid (IPPA) during a symptom-limited stress test. Radionuclide studies were performed after 1 week of placebo treatment (baseline), 4 weeks after oral treatment with 50 mg of gallopamil t.i.d. and again after 1 week of double-blind treatment with gallopamil or placebo. As compared to baseline, initial (poststress) uptake of both tracers in poststenotic myocardial segments was significantly improved after 4 weeks of treatment with gallopamil [thallium-201, +9.0%; p < 0.001; 95% confidence interval (CI), 4.3-13.6%; IPPA, +11.8%; p = 0.003; 95% CI, 4.2-19.3%]. Poststenotic IPPA-clearance was likewise significantly increased (+28.2%; p < 0.001; 95% CI, 12.4-44.0%) indicating a considerably enhanced myocardial fatty acid oxidation after treatment. In the final double-blind phase, myocardial uptake of both tracers as well as IPPA clearance remained enhanced in the subgroup of patients receiving gallopamil and returned to baseline values in patients receiving placebo. Thus, in poststenotic myocardium, chronic treatment with gallopamil provokes an improvement of both regional myocardial perfusion (as demonstrated by an increased tracer uptake in poststress scintigrams) and regional myocardial fatty acid utilization (as demonstrated by an increased uptake and clearance of IPPA).


Diastolic ventricular dysfunction in noncomplicated acute myocardial infarction: the influence of gallopamil.

The goal of this study was to assess the effect of gallopamil on left ventricular (LV) diastolic function early after acute myocardial infarction (AMI). Gallopamil was compared with placebo and atenolol in two different groups of patients. Study patients, 2 days after experiencing their first AMI, in Killip class I and stable sinus rhythm, were randomized in a crossover, double-blind sequence to receive (a) gallopamil (50 micrograms/kg over 5 min) or placebo (i.v. 10-ml bolus, with a time interval of 90 min); and (b) gallopamil (50 micrograms/kg over 5 min) or atenolol (5 mg over 5 min), with a time interval of 24 h. Group I and group II consisted of 28 patients (26 men and 2 women; mean age 55 +/- 9 years) and of 14 patients (13 men and 1 woman; mean age 56 +/- 10 years), respectively. All the patients were treated with thrombolysis within 6 h from the onset of symptoms. Doppler echocardiographic examinations were performed as follows: at baseline (B) and 15 min after administration of gallopamil bolus and placebo bolus, in group I; at baseline before gallopamil (BG) and atenolol (BA), and 15 min after each bolus in group II. The following echo-Doppler parameters were calculated, at each examination: the early and late transmitral peak velocity ratio (E/A), the early and late velocity integral ratio (Ei/Ai), the peak filling rate normalized to mitral stroke volume (nMPFR), and the LV isovolumic relaxation time (IVRT), which was normalized to an 800-ms R-R cycle length in the patients in group II. Mean blood pressure (MBP) and heart rate (HR) were also measured.(ABSTRACT TRUNCATED AT 250 WORDS)


Exercise-induced symptomatic and asymptomatic myocardial ischemia in patients with severe coronary artery disease: focus on the efficacy and safety of gallopamil.

Symptomatic and asymptomatic episodes of transient myocardial ischemia are well-known risk factors in patients with coronary artery disease. In a single-blind, randomized, and placebo-controlled study, the efficacy and safety of gallopamil was studied during a 1-week treatment period in 25 patients with high-grade coronary artery stenosis and frequent, exercise-induced episodes of myocardial ischemia. Eighteen patients were men, and seven patients were women; the mean age +/- SD was 59 +/- 7 years. After a 1-week run-in period (days 1-7), all patients were treated with gallopamil 50 mg t.i.d. (days 8-14) and placebo t.i.d. (days 15-21) or vice versa. Twenty-four-hour Holter monitoring, exercise testing, and adverse effects were controlled at days 7, 14, and 21. During the run-in period, all patients suffered a mean of 5.9 +/- 2.9 episodes of transient myocardial ischemia, mean ischemic duration was 38 +/- 29 min/day. Gallopamil increased exercise tolerance from 7.9 to 9.8 min (+24%, p < 0.05) and resulted in reduction of the weekly usage of short-acting nitrates by 45% compared to placebo. During 24-h Holter monitoring, mean heart rate at the onset of ST-segment depression increased from 106 to 118 beats/min (p < 0.05). The frequency of daily ischemic episodes was reduced after gallopamil administration from 6.1 to 3.9 episodes/day (-37%, p < 0.05), the total ischemic burden decreased for symptomatic episodes by -54% (p < 0.05) and for asymptomatic episodes by 29% (p < 0.05). Gallopamil modified the circadian distribution of ischemic episodes by modifying the morning peak of transient myocardial ischemia.(ABSTRACT TRUNCATED AT 250 WORDS)


Combined gallopamil and isosorbide-5-mononitrate in "mixed" angina pectoris.

The efficacy of combining gallopamil and isosorbide-5-mononitrate (IS-5-MN) was evaluated in 15 patients with "mixed" angina and documented coronary artery disease who participated in a 4-week, double-blind, double-dummy, crossover, placebo-controlled trial. After the first week of the placebo phase (single-blinded), all patients received in three different weeks IS-5-MN 20 mg three times daily, gallopamil 50 mg three times daily, and the same dosages of IS-5-MN and gallopamil three times daily. Exercise tolerance, and peak values of heart rate, systolic blood pressure, double product (DP/100), and ST-segment were evaluated with a treadmill test at the end of each phase. The improvement in exercise tolerance obtained by the combination of the two drugs was significantly greater (p < 0.01) than that achieved by IS-5-MN but not that by gallopamil monotherapy (NS). This effect was accompanied by significant (p < 0.05) reduction (-61%) in ST-segment and significant (p < 0.05) increment (+8%) in peak heart rate only after administration of the combination of the two drugs. The number of ST-depression (ST-) > 1 mm or ST-elevation (ST+) episodes on 24-h Holter monitoring lasting > or = 1 min were also noted in all patients at the end of each phase of the trial.(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical evaluation of calcium-antagonist drugs.

Calcium-antagonist drugs are therapeutic agents of first choice in patients with coronary artery disease. We have reviewed a number of clinical trials in which the safety and efficacy of calcium blockers have been tested and discuss the established clinical effects of these compounds, which range from relief of angina and improved quality of life (both in patients with ischemia due to reduction in coronary flow and in patients with ischemia due to increased O2 demand) to a favorable effect on the course of coronary atherosclerosis and, finally, (at least for some of these agents) to an improvement in prognosis.


Slow-release gallopamil in patients with stable effort angina.

Gallopamil (GSR) is a new calcium-channel blocker. The anti-ischemic activity of GSR was investigated in 12 patients with stable angina of effort, with fixed ischemic threshold (variations < 15%). After a 7-day washout period, patients were randomized to receive treatment with either GSR 100 mg or placebo twice daily for 7 days. Patients underwent maximal symptom-limited exercise test, 10 W/min on a bicycle, during washout (twice) and after the end of each treatment period. Patients were studied by electrocardiogram and the cuff method for determining systolic blood pressure. After treatment with GSR, ischemic and anginal thresholds were increased for at least 12 h in comparison with placebo (ischemic threshold: GSR 663 +/- 37, placebo 571 +/- 36, p < 0.01; anginal threshold: GSR 708 +/- 32, placebo 646 +/- 38, p < 0.05). Rate-pressure product was not changed at the same levels of exercise, but it was significantly increased during exercise at ischemic threshold. In conclusion, GSR possesses an anti-ischemic and antianginal activity lasting at least 12 h. This activity seems due to an increase of coronary blood flow to ischemic areas.


Slow-release gallopamil evaluated by exercise test and long-term electrocardiography.

The effects of slow-release gallopamil (100 mg b.i.d.) were studied on exercise-induced ST-segment depression as well as on spontaneous myocardial ischemia detected by long-term electrocardiography (ECG) monitoring for 48 h in 26 patients with coronary artery disease and angina pectoris. Eight patients had to be excluded (because of paroxysmal atrial fibrillation in four patients, development of unstable angina pectoris in three patients, and frequent ventricular premature beats in one patient). In the remaining 18 patients, gallopamil led to an increase of work load (W x min) evaluated by bicycle ergometry, paralleled by an increase of exercise duration until the occurrence of ST-segment depression of > or = 0.1 mV in the nonblinded part of the trial. The number of spontaneous episodes of myocardial ischemia during long-term ECG recording, ranging from 0 to 14 during control, decreased in patients with two or more episodes during control, paralleled by a decrease in the total duration of ischemic episodes and a decrease in the ischemic score (duration of episodes x maximal ST-segment depression). During long-term ECG monitoring, we observed asymptomatic episodes of spontaneous second degree atrioventricular block of the Wenckebach type in three patients. No other adverse effects of slow-release gallopamil were observed. Therefore, these preliminary results of the non-blinded protocol confirm the anti-ischemic effects of slow-release gallopamil given 100 mg b.i.d.; however, these promising results will have to be confirmed in the consecutive double-blind, placebo-controlled part of the trial.


Efficacy and tolerability of slow-release gallopamil in patients with stable exercise-inducible angina pectoris.

The anti-ischemic properties and tolerability of a slow-release formulation (SR) of gallopamil were investigated in 118 patients with exercise-inducible ST-segment depression and stable angina pectoris in this double-blind, randomized, placebo-controlled, multicenter study. After a placebo run-in period (A) of 2-7 days and a 7-day open therapy period (B) with gallopamil SR, the patients were randomized to a double-blind 7-day period (C) to receive placebo or gallopamil SR 100 mg twice a day. Each patient was submitted to gradual upright bicycle ergometry and electrocardiography (ECG) at rest on the last 2 days of each period at 6 and 12 h postadministration (p.a.) In period C, exercise time and exercise tolerance remained significantly prolonged at 6 and 12 h after gallopamil SR administration in comparison with the placebo values. Additionally the sum of ST-segment depression and maximal ST-segment depression were significantly reduced by gallopamil SR at 6 h p.a. as were the frequency of angina attacks and nitroglycerin consumption. Four patients were withdrawn from the study because of gallopamil-related adverse events, which, however, were not serious. Constipation was noted in 2.5% of the patients. These data suggest that gallopamil SR is effective in reducing exercise-inducible ST-segment depression and increasing exercise tolerance with no serious adverse effects in patients with stable angina pectoris.


Demonstration of specific neuronal cell groups in rat brain by beta-galactosidase enzyme histochemistry.

beta-Galactosidase activity as illuminated by the indigogenic X-gal staining method has been used to demonstrate the presence of genetically modified cells carrying the reporter gene lacZ, coding for the E. coli enzyme. Endogenous activity has been assumed to be minimal since the pH optimum for the mammalian enzyme is 3.5-5.5, while the pH optimum for the E. coli enzyme (and thus of the staining procedure usually employed) is 7.3. Background staining has been reported to be limited to pericytes and a few specific neuronal cell groups. In contrast, our investigations of normal rat brain anatomy demonstrate that many specific neuronal cell groups possess endogenous beta-galactosidase activity when staining is performed at physiological pH. This suggests that background staining of endogenous beta-galactosidase activity in the rat brain has been underestimated. In addition, such specific activity would afford an additional means of identification and illustration of these cells.


Voltammetric monitoring of brain extracellular levels of serotonin, 5-hydroxyindoleacetic acid and uric acid as assessed by simultaneous microdialysis.

We have previously developed a microcomputer-assisted curve-fitting method for measuring the components of the mixed electrochemical signals recorded by differential normal pulse voltammetry in the living brain. It was initially used for resolution of the dopamine and dihydroxyphenylacetic acid components of the catechol signal (peak 2). This report shows how it can be applied to analysis of the indoleamine/uric acid (UA) components of the more complex peak 3. The voltammogram is modeled as a mixture of 3 normal curves of known parameters corresponding to the oxidation of UA, 5-hydroxyindoleacetic acid and serotonin, which is solved by non-linear iterative procedures. Performance was assessed by treatments with drugs having well-known effects on the substances monitored, pargyline and allopurinol, and by the chromatographic analysis of microdialysates collected simultaneously from the contralateral side.


HRP reacted with the chromogen o-tolidine produces whole-cell reaction product at light and electron microscope levels: negative effects of sucrose and Golgi staining on benzidine reactions.

Studies of pathway microcircuitry often require electron microscope analysis. To facilitate these analyses, methods for labeling cells in their entirety are extremely useful. Furthermore, such a method would be most useful if the label would be completely confined by the cell membrane so that second labels for synapse identification could be used. No existing method reliably and repeatably produces this kind of a result. In seeking to develop such a method, a seldom-used chromogen for horseradish peroxidase (HRP) was found which produced superlative results for light and electron microscope analysis. o-Tolidine (3,3'-dimethylbenzidine) reacted with HRP produces a very electron-dense reaction product distributed uniformly throughout the cytoplasm and nucleoplasm; membranes are unobscured so that mitochondria, lysosomes, Golgi apparati and endoplasmic reticula are well defined. The reaction product extends into cellular processes of all sizes, including processes with cell bodies not within the plane of section, and is easily visualized at even the lowest electron microscope magnification. The HRP reaction product is completely confined by the cell membrane, thus terminals presynaptic to labeled cells remain distinct. However, the o-tolidine/HRP reaction product is negatively affected by exposure to oxidizers. In tissue exposed to sucrose before or after being reacted with o-tolidine, the HRP reaction product is less electron dense and is found only in lysosomes outside the nucleus or occasionally in proximal cellular processes. The o-tolidine/HRP reaction product is similarly affected when exposed to potassium dichromate for Golgi staining. For some other benzidine compounds, the chromogen/HRP reaction product is also negatively affected by exposure to these chemicals. Therefore, o-tolidine is a superior chromogen for HRP, labeling cells with details similar to that found for cells intracellularly injected with HRP.


Protective effect of zinc acexamate on experimental gastric ulcers: a histochemical study.

The antiulcerogenic effect of zinc acexamate on gastric ulcers induced by reserpine and changes in the morphology of gastric mucosa were studied in rats by histochemical methods. Histochemistry revealed that zinc acexamate preserved reserpine-depleted neutral and acid glycoproteins. ATPase reaction remained strong in nearly normal periglandular capillaries. The reaction intensity of SDH and NADH2-tetrazolium reductase, and the number and size of the DH-positive parietal cells were decreased, illustrating the decline of energy metabolism involved in acid secretion. The decreased height and weaker staining of the pyroninophile chief cell layer corresponded to the lower amount of RNA, an indirect indicator of pepsinogen synthesis. The significant correlation indices "r" between the severity of gastric lesions and histochemical parameters of the defensive (glycoproteins and microvascular ATPase) and aggressive factors (parietal cell DH and chief cell RNA) confirmed the pathogenic effect of reserpine and the protection provided by zinc acexamate. These findings confirm the multifactorial mechanism of action described for zinc acexamate in several previous works.


Metaphor as illness: postmodern dilemmas in the representation of body, mind and disorder.

The search for a synthesis bridging the gap between materialist and idealist approaches in anthropological theory has been invigorated by recent efforts to develop a critical medical anthropology. Not limited to integrating class analysis and cultural interpretation, the "mindful body" paradigm also aims at empowering the ill, whose experience is denied by biomedical and psychiatric categories that locate disease either in the body or in the mind, and treat them separately from one another and independently of social context. However, missing from the "mindful body" discourse is a reflexive awareness of its contextual grounding in both popular and biomedical discourses of illness, with which it exchanges meanings and from which it borrows dominating power. The case of cancer as a metaphorized illness and a pathologized trope is used to illustrate this process.


The effect of aflatoxin B1 exposure on serotonin metabolism: response to a tryptophan load.

Semi-chronic exposure of ICR male Mice to Aflatoxin B1 (AFB1) in non-toxic doses decreased brain serotonin (5-hydroxytryptamine, 5-HT), 5-hydroxyindole-3-acetic acid (5-HIAA) and catecholamines without altering tryptophan (TRP) or tyrosine (TYR) levels. A TRP load (300 mg/kg, i.p. x 2 hours) slightly increased brain TRP levels while causing a slight decrease in 5-HT and 5-HIAA in control animals. A TRP load in AFB1 treated mice increased brain 5-HT and 5-HIAA. The TRP load caused a further reduction in brain catecholamines without altering TYR levels. Exposure to AFB significantly increased lung TRP levels without altering 5-HT or 5HIAA levels. TRP loading increased lung TRP concentrations in control mice. However, in AFB1 treated mice the increase was not significantly elevated above the level caused by AFB1 treatment alone. Lung 5-HT or 5-HIAA levels in control or AFB1 treated mice are not significantly altered by TRP loading. These experiments demonstrate that AFB1 alters brain and lung TRP metabolism.


Effect of vitamin C administration in modulating some biochemical changes in arthritic rats.

Two phases of arthritis, acute phase (four days after adjuvant inoculation) and chronic phase (21 or 29 days after adjuvant inoculation) were studied in male rats. The effect of administration of vitamin C in a daily oral dose of 50 mg/kg body wt for four and 21 days starting on the day of adjuvant inoculation and for 7 days starting 21 days after adjuvant inoculation against these phases of arthritis was demonstrated. Results showed that prolonged administration of vitamin C (21 days) increased the lowered serum sulphydryl (SH-groups) to prearthritic values while it decreased the elevated level of blood glutathione (GSH) of arthritic rats. However, neither (four-day) nor seven-day treatment with vitamin C exerted any significant changes in these parameters. The results showed also a slight significant increase in the level of erythrocyte superoxide dismutase activity (SOD) [1.15.1.1] upon seven-day treatment with vitamin C. Meanwhile, four-, 21- or seven-day treatment with vitamin C produced no significant change in the elevated levels of serum ceruloplasmin (Cp) and alpha 2-macroglobulin (alpha 2-M) of arthritic rats. However, 21-day and 7-day administration of vitamin C has improved the lowered A/G ratio in these animals. The improvement in these parameters after prolonged administration of vitamin C was explained in the light of the antioxidant property of this vitamin and suggests a beneficial role for it in the treatment of arthritis.


[Adhesion of lymphocytes to allogenic endothelium. Four different pathways].

Lymphocyte adhesion to allogeneic endothelium is a critical step in graft rejection. To further characterize the expression and the regulation of adhesion molecules involved in this process, we stimulated endothelial cells with Interleukin 4 or tumor necrosis factor alpha (TNF alpha) and blocked lymphocyte adhesion to stimulated-endothelial cells with monoclonal antibodies. We demonstrated that lymphocytes bound endothelial cells by at least four adhesion pathways: a) a LFA 1/ICAM 1 dependent pathway on TNF alpha-stimulated endothelial cells, b) a LFA 1 dependent/ICAM 1 independent pathway on unstimulated or IL 4-stimulated endothelial cells, c) a VLA 4/VCAM 1 dependent pathway on unstimulated and stimulated-endothelial cells, d) a CD 2 dependent/LFA 3 independent pathway on IL 4 and TNF alpha-stimulated endothelial cells.


[T CD57+ lymphocytes in kidney transplanted patients. Phenotypic and functional study].

Phenotypic analysis showed a significant increase of CD3+/CD57+ lymphocyte subpopulation in 88 kidney-transplanted patients when compared with non-transplanted patients. No clear correlation was found between the increase of this cell subpopulation and renal function, immunosuppressive regimen, and allogenic or viral stimuli, when each of these factors was considered. Conversely, there was a significant correlation between the proportion of CD3+/CD4+/CD57+ cells and the post-transplant follow-up period. Finally, functional study of CD3+/CD57+ cells suggested that they have suppressive properties, in vitro.


[Amplification of a circulating CD4+CD7- T lymphocytes subset in kidney transplantation].

A minor subset of CD4+ T-cells (10 +/- 4 percent) that does not express the CD7 antigen is detectable in normal individuals. These CD4+CD7- T-cells have lower proliferative capacities than autologous CD4+CD7- T-cells. We observed an expansion of CD4+CD7- peripheral T-cells in 45 kidney transplant recipients. The CD4+CD7- TL expansion (23 +/- 20 percent was observed in both short term (< 6 months, n = 20) and long term transplanted patients (> 3 years, n = 25). These results significantly differed from those observed in patients on dialysis (10 +/- 4 percent, P < 0.002). The percentages of CD4+CD7- TL in kidney recipients are significantly correlated with the percentages of CD4+CD57+ and CD8+CD57+ T-cells subsets. A three-color immunofluorescence analysis indicated that 75 percent of the CD4+CD7- TL express the CD57 molecule contrasting with those in normal individuals (0.3 percent). Purified CD4+CD7- T-cells from kidney recipients have lower proliferative responses after PHA, CD2 and CD3 stimulations than autologous CD4+CD7+ TL. The proliferative response is partially restored by the addition of anti-CD28 moAb or IL2. We have highlighted, in kidney recipients with good graft function, an expansion of CD4+CD7- T lymphocytes which could be associated with CD4+ T-cell anergy and tolerance.


Effect of benzodiazepines on tryptophan binding to rat hepatic nuclei.

This study evaluates whether or not some of the benzodiazepines would influence the binding of L-tryptophan to rat hepatic nuclei or nuclear envelopes. Previous publications have indicated that binding of L-tryptophan to hepatic nuclear envelope proteins was saturable, stereospecific, and of high affinity. In this study, we investigated whether some of the benzodiazepines would influence L-tryptophan binding to rat hepatic nuclei or nuclear envelopes as assayed by in vitro L-(5-3H) tryptophan binding. Our results indicate that the addition of chlordiazepoxide, diazepam, prazepam, flurazepam, nordazepam, N-desalkylflurazepam, temazepam, oxazepam, lorazepam, or 4-chlorodiazepam has little influence on the L-(5-3H) tryptophan binding to hepatic nuclei in vitro. However, the addition of demoxepam, the N-desalkylated compound of chlordiazepoxide, caused marked competition with 3H-tryptophan binding to hepatic nuclei in vitro. When chlordiazepoxide (1 mg/100 g body weight) is administered intraperitoneally 20 min before killing, the isolated hepatic nuclei reveal decreased specific L-tryptophan binding compared to controls. Also, rats pretreated with chlordiazepoxide intraperitoneally before tube-feeding L-tryptophan revealed diminished tryptophan-induced hepatic nuclear RNA efflux and protein synthesis. Our results suggest that chlordiazepoxide, possibly by itself or through a metabolite, can act to affect hepatic nuclear binding of L-tryptophan and to inhibit the stimulatory effect of L-tryptophan on hepatic protein synthesis.


N-nitrosoheptamethyleneimine-induced pulmonary and esophageal carcinogenesis and effects of concomitant treatment with bleomycin in rats.

The combination effects of bleomycin with N-nitrosoheptamethyleneimine (NHMI) or dihydroxy-di-N-propylnitrosamine (DHPN) on pulmonary carcinogenesis were investigated. Male F344 rats were given NHMI (20 or 40 ppm) or DHPN (200 ppm) in the drinking water and intraperitoneally injected with bleomycin (1 mg/kg) once a week for 18 weeks and then killed at week 24. Many rats treated with NHMI died before the termination of the experiment due to toxicity or development of advanced esophageal carcinomas, considered to be the main cause of death. Detailed histological examination performed on rats killed at week 24 revealed no statistically significant effects of bleomycin on NHMI or DHPN induction of neoplastic lesions in the lung or esophagus, although pulmonary carcinomas were only found in two rats treated with NHMI plus bleomycin. Under the present experimental conditions, NHMI exerted stronger carcinogenic activity in the esophagus than in the lung, and no obvious modifying effects of simultaneously administered bleomycin were evident on NHMI- or DHPN-induced pulmonary carcinogenesis.


[A proposal of the essential elements for teaching the process of nursing care and its application to the surgical patient].

Greater dynamism is advocated in the teaching-learning process of Nursing Care by proposing essential skills which are both logical and specific, necessary and sufficient, to provide people (healthy or diseased), the family and the community with more efficient and quicker nursing cares. The theoretical foundations of the invariants proposed are stated and a description is given of the operational method for evaluating students' case discussions by using a measuring scale in a surgical patient with hydric, electrolytical and acid-basic unbalance. The results reached are satisfactory since 50% of the students were evaluated as "very high". Continuation of the study with other groups of students is recommended.


[Changes in the blood glucose of revascularized patients in the immediate postoperative period].

Variability in glycaemia in revascularized patients in the immediate postoperative period was studied in order to determine the presence of hyperosmolar hyperglycaemia coma, ketogenic and non-ketogenic, and the relation with hemodynamic changes. It was found that 15 patients (9.2%) from the Arteriology Service and two patients (7.1%) from the Diabetes Angiopathy Service had a non-ketogenic hyperglycaemic hyperosmolar coma and there was no ketogenic hyperglycaemic hyperosmolar coma. There were 32 patients (16.8%) with hemodynamic changes who had mild and moderate hyperglycaemia in the immediate postoperative period.


Severe cutaneous hypersensitivity reactions during treatment of tuberculosis in patients with HIV infection in Tanzania.

Concurrent infection with HIV-1 and Mycobacterium tuberculosis is increasingly common in East Africa. In the past, a drug regimen consisting of 2 months of intramuscular streptomycin plus 12 months of isoniazid and thiacetazone has been used in tuberculosis control programs with acceptable efficacy and low incidence of adverse reactions. Anecdotal reports of increasing cases of Stevens-Johnson syndrome prompted a 2 month prospective search for cases of severe cutaneous hypersensitivity reactions at Muhimbili Medical Centre in Dar es Salaam, Tanzania. Five such patients were admitted to a single ward during this time, 4 of whom were HIV-seropositive and all of whom were being treated with isoniazid and thiacetazone. These findings have implications for the management of tuberculosis in East Africa and perhaps other countries with high prevalence of both HIV-1 and tuberculosis.


[The mapping of the neurons that participate in the innervation of the body wall of the edible snail].

In the paper are presented the results of the experiments with the retrograde staining of the left-side cutaneous nerves of the pedal ganglia of the snail Helix lucorum. The somata of neurons which sent processes forming the studied nerves were revealed with the help of nickel and cobalt ions precipitation by the rubeanic acid. A lot of nerve cells including some command neurons were found in all the ganglia excluding the buccal ones. Especially large amount of neurons was stained in the ipsilateral pedal, pleural, and parietal ganglia.


[The structural organization of the snail sensory systems].

In the paper are reviewed the author's data on the structure of peripheral nervous system of the body wall, gravitational organs--statocysts, eyes, and organs of distant chemoreception--ommatophors, or posterior head tentacles, of the snails Helix vulgaris and Helix pomatia. Localization in the CNS of central parts of the main sensory systems is shown and some structural characteristics of them are described. The data on the organization of the procerebrum which takes part in processing of information from the receptors of the head tentacles are presented. By some morphological features the procerebrum may be considered as one of the highest associative centers of the snails. The data under consideration were obtained by means of silver impregnation by Golgi, retro- and anterograde infusion of CoCl2, horseradish peroxidase and Lucifer yellow into the nerves, as well as by means of a number of other classical histological techniques.


[The facilitation of defensive reactions during food consumption in the snail helix: the participation of glucose and gastrin/cholecystokinin-like peptide].

Molecular and cellular mechanisms of the interrelations between the feeding and defense behaviour were studied in a snail Helix lucorum. The dynamics of defense reactions was investigated in snails with different levels of feeding motivation. Defense reactions were suppressed in hungry snails, while 15-20 min after the beginning of food intake they were facilitated. The facilitation depended on a duration of starvation. Injection of 0.5 ml of 5 mM glucose solution (up to the glucose level in the haemolymph of a food satiated snail, 1.6-2.0 mM) or injections of 20-30 ng of synthetic analogues of the gastrointestinal peptides (pentagastrin of octapeptide cholecystokinin, CCK-8) facilitated the defense reaction in a hungry snail. Parameters of the facilitation were similar to those in the period of food intake. Activity of the command neurons of defense behaviour (L-PPL1) after the carrot juice application to the lip of a semi-intact preparation from a hungry snail was glucose-dependent. Similar glucose-dependent changes of L-PPL1 activity were found after CCK-8, but not FMRFamide application during the perfusion with 0.5 mM glucose. L-PPL1, but not L-PPa2-3 neurons were most sensitive to glucose and CCK-8 level changes in the Ringer solution. Adaptive significance of the behavioural phenomena as well as glucose and gastrin/CCK-like peptide participation in these processes are discussed.


[The cholinoreceptors of the neurons in the edible snail: their identification and plasticity and its regulation by opioids and second messengers].

In the paper is presented the review of author's own data on the cholinoreceptors of identified Helix neurons and the regulation of their plasticity. The review contains the following parts: cholinoreceptors identification, their coupling with ionic channels, identification of opioid receptors, modulation of cholinoreceptors by opioids, plasticity of cholinoreceptors, modulation of their plasticity by the second messengers, second messengers involved in a modulation of cholinoreceptors' plasticity by the opiate kappa-agonist bremazocine.


Induction of mucosal and serum immune responses to a specific antigen of periodontal bacteria.

The dynamics of the host immune response to periodontal bacteria not only may be informative from the standpoint of specific mucosal protection to these pathogens, but also may reveal the capacity of the mucosal immune response to provide protection of the host. To this end, we have examined the immune response to chromatographically purified fimbriae of P. gingivalis administered orally or systemically with liposomes and adjuvant in BALB/c mice, high responders to this antigen. Oral administration of P. gingivalis fimbriae clearly enhanced the fimbriae-specific salivary IgA response. ELISPOT analysis revealed that significant numbers of fimbriae-specific IgA SFC were seen in lamina propria and mesenteric lymph nodes but not in Peyer's patches of mice immunized orally. In contrast, antigen-specific IgM and IgG SFC were seen mainly in the circulating blood mononuclear cells. On the other hand, subcutaneous injection of fimbriae with GM-53 also raised the fimbriae-specific IgG followed by IgM and IgA responses in serum, and both IgA and IgG responses in saliva. Oral immunization was less effective than subcutaneous injection in terms of the serum antibody response. However, the salivary antibody level of mice injected subcutaneously was similar to that of mice immunized orally. In the subcutaneously immunized mice, fimbriae-specific SFC were detected in the spleen, blood, and brachial lymph nodes by ELISPOT assay. Fimbriae-specific IgM SFC appeared earlier and antigen-specific IgG SFC were seen later. These results show that the combined use of fimbriae together with the adjuvant results in sharply increased IgA responses in saliva and IgG responses in serum. In summary, it is clear that the nature of the host's antibody response in serum and mucosal secretions is distinct, and depends on the route of antigen administration, the use of adjuvant and/or liposomes, and the temporal phase of the humoral immune response following various immunization regimes.


Canatoxin triggers histamine secretion from rat peritoneal mast cells.

Canatoxin, a toxic protein present in the seeds of Canavalia ensiformis, induces the secretion of serotonin, dopamine and insulin through activation of the lipoxygenase pathway. The purpose of the present study was to verify if canatoxin causes histamine release from rat peritoneal mast cells and to perform a detailed study of this phenomenon. Our results indicate that canatoxin is capable of activating mast cells to release histamine. The process is time- and concentration-dependent, occurs without cell damage and requires metabolic energy as well as the presence of divalent cations (Ca2+ and Mg2+). Optimal release occurs at 37 degrees C and at physiological pH. Extremes of temperature (0 degree C and 45 degrees C) inhibit the process. We conclude that canatoxin induces histamine release from rat peritoneal mast cells by an active secretory process.


The effects of histamine, pyrilamine, cimetidine, and ranitidine on secretion of lingual lipase and amylase from rat von Ebner's glands.

Minced von Ebner's glands of rat tongue were incubated in vitro with histamine and histamine receptor antagonists. At various time intervals, media and homogenates of the tissue were assayed for lingual lipase and amylase activity and percentage secretion calculated. Histamine elicited moderate secretion (approximately 10%) of lingual lipase and amylase. In contrast, pyrilamine, an H1 receptor antagonist, elicited > 60% secretion. There were statistically significant differences between the percentage secretion of lingual lipase and amylase for basal secretion, as well as for histamine- and pyrilamine-evoked secretion above basal. The H2 receptor inhibitors, cimetidine and ranitidine, stimulated secretion of only amylase, but not lingual lipase. When combined with histamine, these antagonists partially inhibited only the secretion of histamine-evoked lingual lipase, but not amylase. The differences in percentage secretion between the two enzymes indicate that exocytosis may not be the only process involved in protein secretion. The anomalous effects of the H1 and H2 receptor antagonists necessitate a more detailed characterization of the receptors of von Ebner's glands.


Substance P as a potent stimulator of sneeze responses in experimental allergic rhinitis of guinea pigs.

Substance P was examined for sneeze-inducing activity and its involvement of sneeze responses in experimental allergic rhinitis. Substance P, dripped into a nostril of guinea pigs, at concentrations of 100 pM and above induced sneezing in a dose-dependent fashion. The activity of substance P was not affected by the previous subcutaneous injections of capsaicin that depleted substance P in nerve fibers. Histamine induced sneezing at concentrations of 30 mM and above and the activity was reduced by capsaicin treatment. The frequency of antigen-induced sneezing was proportional to the substance P content in nasal mucosa of sensitized guinea pigs treated with increasing doses of capsaicin; correlation coefficient 0.91. These results suggest that substance P plays an important role as a stimulator of sneeze responses in experimental allergic rhinitis in guinea pigs.


Suppressive effects of E3330, a novel quinone derivative, on tumor necrosis factor-alpha generation from monocytes and macrophages.

E3330 [(2E)-3-[5-(2,3-dimethoxy-6-methyl-1,4-benzoquinoyl)]-2-nonyl-2- propenoic acid], a novel synthesized hepatoprotective compound, has suppressive effects on tumor necrosis factor-alpha (TNF-alpha) generation from monocytes/macrophages in vitro. E3330 (1-100 microM) reduced lipopolysaccharide (LPS, 10 mg/ml or 1 microgram/ml)-induced TNF-alpha generation from rat resident and Propionibacterium acnes (P. acnes)-elicited peritoneal macrophages, rat and human monocytes, rat Kupffer cells, and splenic mononuclear cells in a concentration-dependent manner. E3330 also (1-100 microM) suppressed TNF-alpha generation stimulated with egg-albumin immune complex in rat P. acnes-elicited peritoneal macrophages. Northern blot analysis showed that LPS-induced expression of TNF-alpha messenger RNA (mRNA) in human blood monocytes was suppressed by E3330. These findings indicate that E3330 has a suppressive effect on TNF-alpha generation from monocytes/macrophages, regardless of origin or species, and this effect is based in part on the suppression of TNF-alpha mRNA expression.


Knowledge about developmental screening in mothers attending a maternal and child health clinic in Singapore.

A study of knowledge about Developmental Screening in mothers was done among 200 Singapore mothers attending a Maternal and Child Health Clinic from February to March 1990. They were requested to answer a set of questions pertaining to their knowledge on the Developmental Screening session and their reason for attending the session. They were also asked whether they had attended any health education talks given by the nurses in the clinic. Seventy-three out of 200 (36.9%) knew what the appointment given was for. Mothers with secondary education were almost three times and mothers with tertiary education were five-and-a-half times more likely to possess the correct knowledge than a mother with primary education. Wives of husbands from Social Classes I and II were more knowledgeable (83% and 54%) than those in lower social classes (32% and 25%). Mothers with older children were more knowledgeable than those with children age six weeks. Mothers of one child four-and-a-half years old were more knowledgeable than those with one child age six weeks. Twelve out of 27 (44%) of mothers of one child age six weeks did not know what Developmental Screening was about whereas all mothers with four-and-a-half year old children knew what Developmental Screening was about. Only 88 (44%) mothers had attended health education talks. There was no discernible difference in knowledge of Developmental Screening between those mothers who attended talks and those who did not (35% vs 40%).(ABSTRACT TRUNCATED AT 250 WORDS)


Renal tubular function of cadmium exposed workers.

The kidney is considered the critical organ following long term occupational or environmental exposure to cadmium. Tubular dysfunction in the form of low molecular weight proteinuria is the earliest manifestation of cadmium nephrotoxicity. The current acceptable critical concentration of cadmium in the urine is 10 ug Cd/g creatinine. The aim of this paper is to identify the presence of tubular dysfunction among workers with less than 10 ug Cd/g creatinine. The exposed group of 92 workers were from a nickel-cadmium battery factory. The control group of 122 workers were factory and sedentary office workers with no known history of exposure to nephrotoxic agents. The urinary excretion of N-acetyl-D-glucosaminidase (NAG), beta-2-microglobulin (beta 2m) and alpha-1-microglobulins (alpha 1m) were measured from morning spot urine samples. The age, sex and race adjusted NAG and alpha 1m showed increasing trend with rising urinary cadmium levels. Levels were significantly raised when the urinary cadmium was above 5 ug Cd/g creatinine. A similar trend was seen with increasing length of exposure. Renal tubular dysfunction is present among cadmium exposed workers with levels below the current critical concentration.


Vimentin expression in benign and malignant lesions in the human mammary gland.

The distribution pattern of vimentin in mammary gland lesions, including benign lesions (36 cases) and adenocarcinomas (40 cases), was studied with the use of a monoclonal antibody. In myoepithelial cells of mastopathy, the positive vimentin staining was seen in 6 cases (33.3%). Gynecomastia showed strong vimentin expression in myoepithelial cells. Fibroadenomas (14 cases) expressed positive vimentin staining in 8 cases (57.1%) with various staining intensities in myoepithelial cells. Lactating mammary gland tissue was positive for vimentin staining in narrow myoepithelium. In adenocarcinoma, positive reactions in tumor cells were found in 17 cases (41.2%) of solid tubular, 12 cases (33.3%) of papillo-tubular, and 11 cases (54.5%) of scirrhous carcinomas. The staining intensity for vimentin was weak in modified myoepithelial cells and neoplastic cells. Malignant cells in mammary carcinomas with vimentin reactivity usually expressed keratin staining; thus a co-expression of vimentin and keratin was found in mammary carcinomas.


Enhancement by androgen of the angiogenic ability of androgen-responsive Shionogi carcinoma 115.

The effects of various steroids on the induction of angiogenesis by androgen-responsive Shionogi carcinoma 115 (SC115) were investigated by implanting a piece of a SC115 tumor tissue into a rabbit cornea together with a pellet containing each steroid (250 micrograms) and by examining neovascularization from the limbus. SC115 tumor tissues induced neovascularization in 42% corneas in the absence of steroids, and in 31, 63, 80 and 4% in the presence of estradiol-17 beta, progesterone, testosterone and dexamethasone, respectively. Only testosterone significantly increased the percentage of corneas with neovascularization, while dexamethasone significantly decreased it. Testosterone itself induced no neovascularization, and dexamethasone also inhibited neovascularization induced by acidic fibroblast growth factor. The present results suggest that androgen increases the secretion of an angiogenic factor(s) by an androgen-responsive SC115 tumor, while dexamethasone inhibits angiogenesis induced by the angiogenic factor(s).


Immunohistochemistry in stage FIGO III cervical cancer: prognostic value of tumor associated antigens and intermediate filaments.

The prediction of prognosis in cervical carcinomas solely by histological examination is doubtful. Our study was undertaken to evaluate whether Cytokeratin 5D3, Cytokeratin high molecular weight (HMW), Cytokeratin AE1/AE3, Carcinoembryonic Antigen (CEA) and CA 19.9 could have a prognostic value in primarily irradiated stage FIGO III cervical cancer. We investigated 45 pretreatment punch biopsies of cervical cancer stage FIGO III. All patients underwent a combined standardised primary irradiation treatment. In the cases with expression of Cytokeratin AE1/AE3 survival was significantly better than in cases without expression. The expression of CEA also had a significantly positive effect on survival. The best prognosis was found in cases with coexpression of AE1/AE3 and CEA (Wilcoxon p = 0.006, log-Rank p = 0.03). The other investigated tumor associated antigen and intermediate filaments did not correlate with survival. The application of Cytokeratin AE1/AE3 and CEA allows a prediction of prognosis in patients with stage FIGO III cervical cancer undergoing irradiation treatment.


Squamous cell carcinomas, an immunohistochemical and ultrastructural study.

Fourty-seven squamous cell carcinomas (SCCs) were studied by light microscopy, immunohistochemistry and electronmicroscopy. Nineteen percent (9 cases) of SCCs in different locations were immunoreactive for CAM5.2 and three poorly differentiated SCCs did not express cytokeratins 6 and 18. No cases were positive for c-erbB2 protein. p53 protein overexpression was found in malignant cells in 40% of the primary tumours and in 60% of the lymph node metastases. Four poorly differentiated SCCs expressed vimentin and in these cases the tumour cells had accumulations of less dense intermediate filaments in cytoplasm.


Partial characterization of protein tyrosine kinases in human lymphoid cells.

We have examined several aspects of protein tyrosine kinase (PTK)-activity in the cytosolic and membrane fractions of both normal and malignant lymphoid cells. The expression of the PTK-encoding oncogenes fes, abl and src, was investigated with the use of antibodies generated to their respective gene products. These antibodies recognized proteins in all cell types examined, most frequently in both the cytosolic and the membrane fraction. PTKs were partially characterized by FPLC. PTK-activities of column fractions were assayed using a non-radioactive dot blot assay. Cytosolic and membrane fractions showed FPLC patterns with a constant as well as a variable part in both normal and malignant cells, suggestive of PTKs with specialized functions in normal cell growth and transformation. Lastly, using antibodies to phosphotyrosine, we found that cytosolic fractions contained the majority of proteins phosphorylated at tyrosine in all cell types. Normal peripheral blood lymphocytes and B-lymphoma cells showed a great similarity in tyrosine phosphorylation pattern, while in tonsillar lymphocytes a clearly different pattern was found. These methods further characterize PTK-activities in lymphoid cells, and the results give evidence that PTKs in normal and malignant cells have both similar and different aspects.


Chemosensitivity testing of xenografted squamous cell carcinomas of the head and neck region.

Eight squamous cell carcinomas from the head and neck region were established as xenograft lines in nude mice and tested for their sensitivity to the antineoplastic drugs bleomycin and cisplatin. Tumor volume, histology, DNA flow cytometry and mitotic activity were used as parameters. One out of the 8 tumours appeared to be highly sensitive to bleomycin, while three other tumours were sensitive to both bleomycin and cisplatin. These observations are in good correlation with the reported data in patients. All chemosensitive tumours showed regrowth after the cytotoxic drug treatment had been completed. No change was seen in the chemosensitivity of other features of the regrown tumours, not even after repeated exposure to the drugs. Comparison of the tumour volume with the other parameters applied indicated that the tumour volume of squamous cell carcinomas was not always a reliable parameter for testing chemosensitivity, because of the important contribution of keratin to the tumour volume. It is concluded that additional parameters such as histological examination, DNA flow cytometry or mitotic activity are necessary in order to draw reliable conclusion on xenografts with a large avital component. In addition, DNA flow cytometry has proved to be very useful for the rapid screening of drug sensitivity.


MDR hamster cells exhibiting multiple altered gene expression: effects of dexniguldipine-HCl (B859-35), cyclosporin A and buthionine sulfoximine.

An actinomycin D selected, multidrug-resistant (MDR) hamster CHO subline showed strong expression of the P-glycoprotein and sorcin genes together with several other alterations such as a: (i) reduced growth rate, (ii) lowered topoisomerase II, (iii) lowered glutathione-S-transferase-P gene expression, and (iv) the emergence of a 15.5 kDa protein. Besides high resistances to adriamycin, actinomycin D, and vincristine, we observed a lowered sensitivity towards bleomycin, a rather hydrophilic drug usually not involved in P-glycoprotein associated MDR. Moreover, the MDR subline showed a pronounced collateral (enhanced) sensitivity towards the sterically pure dihydropyridine anticancer drug dexniguldipine-HCl (B859-35) preventing its characterization for MDR modulation here. At a non-cytotoxic dose (10 microM) the immunosuppressive cyclic peptide cyclosporin A completely abolished the resistance to vincristine, partially reversed the resistance to teniposide and strongly enhanced the sensitivity towards bleomycin, while not influencing the drug sensitivities of the parental cell line. Buthionine sulfoximine (BSO), an agent depleting cellular glutathione levels, distinctly increased the sensitivity towards teniposide at nontoxic doses (50 microM) exclusively in the MDR subline, while it did not alter vincristine or bleomycin cytotoxicity.


Distribution of neuropeptides in rat pterygopalatine ganglion: light and electron microscopic immunohistochemical studies.

The distribution and quantity of neuropeptides in the rat pterygopalatine ganglion were studied by using complete serial paraffin sections of the ganglion immunostained with antiserum against several neuropeptides. The pterygopalatine ganglion, composed of 4932 +/- 291 (mean +/- SD) neurons, was triangular in shape with a tapering caudal tail. The most commonly found peptide in neurons was vasoactive intestinal polypeptide (VIP) (99.0%), followed by neuropeptide Y (NPY) (54.1%) and enkephalin (10.5%). The rostro-ventromedial and caudal parts of the ganglion where intensely VIP-immunoreactive neurons predominate project to the nasal mucosa, while the rostro-dorsolateral part of the ganglion where NPY-immunoreactive neurons predominate projects to the Harderian gland. The coexistence of VIP/NPY (47.4%), VIP/NPY/enkephalin (6.6%) or VIP/enkephalin (3.9%) in the ganglionic neurons was recognized. Calcitonin gene-related peptide (CGRP)- and substance P-immunoreactive varicosities formed synaptic contacts with the somatic spine or soma, which confirmed that the reflex arch, composed of axon collaterals of trigeminal ganglionic neurons and parasympathetic ganglionic neurons, operates through direct synapses. Enkephalin-immunoreactive varicosities, which were probably derived from parasympathetic preganglionic neurons, also made synaptic contact with the somatic spine.


Morphology and intermediate filament composition of human mammary epithelial cells treated with stable butyrate derivative.

A new stable butyrate derivative monobut-3 was previously shown to inhibit proliferation and promote differentiation in human mammary established cell lines. The present study on monobut-3's effects on mammary epithelial cells cultured from human non-malignant and malignant breast tissues demonstrated pronounced morphological alterations suggestive of cellular differentiation. In addition, some degree of architectural differentiation was also evident in treated primary cultures. Monobut-3 did not affect the expression of vimentin and cytokeratin 18 when assessed in human breast cell lines expressing one or both types of intermediate filaments. However, it did induce expression of cytokeratin 19, characteristic of fully differentiated mammary cells, in one of the two cell lines devoid of this cytokeratin subtype. Furthermore, the network of intermediate filaments was often more largely extended in cells treated with monobut-3 than in untreated ones. These results indicate that monobut-3 can induce subtle changes in intermediate filaments which may contribute to its ability to promote differentiation in human mammary cells.


[Spontaneous motor activity in mice overloaded with iron-dextran].

We studied the spontaneous motor activity and the levels of iron in brain (frontal cortex and striatum) and liver of mice overloaded with iron dextran. Two groups of mice injected intraperitoneally with two different doses of iron and the control group were used. We found a significant increase in the spontaneous motor activity only for the experimental group treated with the highest dose of iron. In frontal cortex and striatum the concentration of iron did not augment in the experimental groups. However, in liver we detected a highly significant rise in the iron content of both experimental groups. Our results suggest that the increase in the spontaneous motor activity observed in mice injected with the highest dose is not directly caused by changes in the iron concentration in the brain.


Determining the neuronal connectivity of Golgi-impregnated neurons: ultrastructural assessment of functional aspects.

The combined light and electron microscopic analysis of Golgi-impregnated neural tissue is a potent tool for determining the connectivity of neural networks within the brain. In the experimental paradigms commonly applied in these studies, the Golgi-impregnated neurons are typically examined as the postsynaptic neuronal components. The structural characteristics and the pattern of distribution of their synaptic connections with other groups of identified neurons are analyzed. Due to the high power of resolution of the Golgi-electron microscopic technique, the ultrastructural analysis of Golgi-impregnated neurons can be expanded to elucidate activity-dependent structural alterations in their cytoarchitecture. These structural alterations can then be correlated under different physiological conditions with changes in the functional efficacy of the subcellular neuronal components.


Quantitative aspects of synapses on Golgi-impregnated neurons.

With the classical Golgi techniques, numerous types of neurons can be distinguished in the cerebral cortex, each with a specific dendritic geometry and pattern of axonal ramifications. In the present review we describe two techniques which allow quantification of synapses on identified neurons: (1) Golgi-rapid impregnation-gold toning-electron microscopy, and (2) Golgi-Kopsch impregnation-gold toning-electron microscopy in combination with staining of the tissue with ethanolic phosphotungstic acid (E-PTA). Both techniques were applied on neurons in the visual cortex of young and adult rabbits. By means of rotating and tilting specimens in the electron microscope, the nondistinctive ultrastructure of obliquely sectioned synapses can be circumvented, leading to precise estimates of asymmetrical vs. symmetrical synapses without complete reconstruction of the neuron.


[Cryopreservation of human erythrocytes with hydroxyethyl starch (HES)--Part 1: The procedure].

Although the efficacy of hydroxyethyl starch (HES) as a cryoprotectant for human red blood cells (HRBC) was demonstrated in 1967, no clinical application has evolved so far. In contrast to glycerol, which has been widely used for this purpose, HES offers the advantage of being nontoxic if administered intravenously. Being also a plasma substitute, HES in combination with HRBC can normalize blood volume as well as oxygen transport in the case of extended blood loss. This may be of importance for autologous blood supply for elective surgery and for disaster treatment plans. For a volume of 40 ml, the optimal combination of important parameters [e.g. cooling rate 200-250 K/min, warming rate 400 K/min, hematocrit 40%, HES concentration 12% (w/w)] has led to a survival after thawing in terms of saline stability of 93 +/- 1% (n = 10). Additionally, the age prior to cryopreservation (0-3 days) had an influence on postthaw survival, but at least 90% survival can be obtained if the samples are washed 3-5 times before freezing.


[Cryopreservation of human erythrocytes with hydroxyethyl starch (HES)--Part 2: Analysis of survival].

Red blood cells (RBC) were obtained from 5 whole blood units by centrifugation and were purified using a simple washing procedure. Hematocrit and HES concentration in the 108-ml samples to be frozen were adjusted to 40% (v/v) and 12% (w/w), respectively. Cooling was performed by submerging into liquid nitrogen using containers to generate a flat sample geometry (3 mm thickness). After thawing in a shaking water bath, HES and free hemoglobin were removed in a simple washing step. To investigate the influence of the resuspension medium, the RBC were transferred into freshly drawn autologous plasma and into Locke's solution. Survival after thawing in terms of saline stability reached 86.3 +/- 2.3%. The cryopreservation procedure caused no major changes with regard to osmotic fragility, 2,3-DPG or intracellular Na+ and K+. ATP was reduced by 16%, but this had completely recovered after 3 h resuspension in autologous plasma. Some morphological changes present after thawing (e.g. stomatocytes, echinocytes) also recovered after 1.5 h. In conclusion, those RBC which survived the preservation process can be considered to be fully viable with regard to the parameters investigated.


[The characteristics of the HIV isolated from HIV-infected persons and AIDS patients on the territory of the CIS].

HIV strains were isolated from HIV-infected patients and AIDS patients in CIS. A total of 81 HIV isolates were obtained. The isolates were identified by using immunofluorescence and enzyme immunoassay, by determining the activity of reverse transcriptase, immunoblot, electron microscopy, polymerase chain reaction. Of the 81 isolates 79 were HIV-1 and 2 HIV-2. The strains differed in their infectivity, the kinetics of virus antigen accumulation, and the spectrum of susceptible cell lines. The viruses isolated may be assigned as two groups: high and low infective. The biological properties of the national HIV isolates were shown to be similar to the prototype HIV strains isolated elsewhere.


[The possibilities of using immunological indices as criteria for determining the stage of HIV infection and the disease prognosis].

The authors studied the immune status of 14 HIV-infected patients, 6 of whom had lymphadenopathy, 4 were diagnosed as having AIDS-related complex and 4, a full-blown AIDS. Analysis of laboratory findings showed that of predictive value are serum levels of immunoglobulin B, a CD4 cell count less than 200, reduced populations of CD20 and CD16 lymphocytes, and a depressed response to pokeweed mitogen. Based on the clinical manifestations and laboratory results, three stages characterizing the immune system in HIV infection have been identified.


[The expert diagnosis of HIV infection by radioimmunoprecipitation].

Using a panel of sera from HIV-infected persons and donors, the authors showed that radioimmunoprecipitation assays compare favourably with immunoblotting assays. With radioimmunoprecipitation, cross reactions were observed between HIV-2 antigens and HIV-2 antibodies, and that the nature of cross reactivity differs from that observed with immunoblotting. Potentials of radioimmunoprecipitation assays as a confirmatory test for use with sera that have given indeterminate results in immunoblotting assays and contradictory results in enzyme immunoassays are examined.


[The factors that affect the results of the serological diagnosis of HIV infection and the screening for anti-HIV].

The laboratory diagnosis of HIV infection and screening for anti-HIV have determined the factors that influence the results of studies performed by ELISA and Western immunoblot. The factors include sensitivity and specificity of the test systems used, storage conditions of and transportation conditions for serum, errors of procedures for reactions, etc. Correlation is between the inactivation of the sera studied and the initial antibody titers during their storage at elevated temperatures. To obtain more valid results of HIV infection serodiagnosis it is necessary to use some test systems differing in the principles of setting up a reaction.


[The methodological aspects in enhancing the efficacy of the epidemiological surveillance of HIV infection].

Using a systems approach, the authors examined the main principles underlying an expert-based system for epidemiological surveillance over HIV infection. The principle objective is to control the processes which precede the occurrence of new infected cases and further spread of HIV infection. Accordingly, the system should address the following: epidemiological diagnosis and prediction of HIV incidence and spread, development of optimal preventive anti-epidemiological measures, research into how the infection spread. This should incorporate a database which is made by the hierarchic principle: the parameters of higher levels include those of lower level. So that the highest level would include two main integral estimates characterizing the risk of HIV from the outside (the first estimate) and the risk of HIV infection (the second estimate).


[Biotechnical systems--new potentials for research in hygiene].

The authors have worded the main assumptions of the present-day trends in hygienic studies based on the feedback biotechnical systems. It is suggested that biocontrol may be exercised on the basis of automated diagnosis of the hygienically significant conditions of the body. Concrete examples of deciding the indicated problems are reviewed. The principal advantage of the approach suggested lies in the perfection of the methodology of hygienic regulations on the basis of a direct effect of self-regulation of the body on the estimation of the concrete body-medium system.


[Reverse transcriptase inhibitors and the therapy of HIV infection].

Azidothymidine (retrovir) and didesoxyinosine, which represent nucleoside agents, are major remedies in the treatment of HIV infection and acquired immunodeficiency syndrome (AIDS). The paper describes the molecular mechanism of their action. It implicates that triphosphates of these nucleosides selectively suppress the activity of reverse transcriptase (RNA-dependent DNA polymerase) of HIV by a termination mechanism. This results in effective inhibition of HIV reproduction and recovery of lymphocyte count and yields marked therapeutical benefits. The new generation anti-HIV agents are nucleoside-based phosphonates which were discovered by Russian investigators in 1987. The agents having a significant anti-HIV activity are low toxic. Emphasis is made on combined therapy of HIV infection, which holds much promise.


[The evaluation of the mutagenic hazard of atmospheric air pollution for an urban population].

The authors describe the results of a genetic monitoring made in three towns of the Ukraine, which differ in the levels of polluted atmospheric air. Spontaneous abortions (SA) were selected as an index for genetic health of the population. The total number of CA, which were detected by primary documentation in the studied towns, the number of SA in early periods of pregnancy, and the incidence of SA, probably, of mutagenic origin, which is determined by multi-variance analysis, are greater in the contaminated towns than in comparatively pure towns. The findings suggest that the achieved level of atmospheric air pollution in some towns of the Ukraine is genetically significant.


[Disordered integration of the immune and nervous systems in exposure to adverse environmental factors].

Exposure to adverse environmental factors can result in the impaired relationships between the immune and nervous systems and hence increase autonomic properties of immunocompetent cells. This impairment may underlie some diseases associated with abnormal immunity. To have knowledge on the ways of restoring the integrative connections between the above systems will help maintain human health, augment its potential as a creator of the noosphere.


[The participation of prostanoids in the realization of the cardioprotective, antistressor and anti-arrhythmic effects of enkephalins].

It has been established that preliminary administration of D-Ala2-Leu5-Arg6-enkephalin, D-Met2-Pro5-enkephalinamide, and D-phenylalanine to experimental animals prevents a stress-induced increase of the content of thromboxane in the blood plasma and myocardium and induces a rise of the level of prostacyclin, PGF2 and PGE in the heart and blood plasma. The authors hold that the changes in the level of prostanoids may mediate cardioprotective and anti-stressor rather than antiarrhythmic effects of enkephalins.


[Pancreatic juice peptides as possible regulators of reparative processes in the pancreas in acute and chronic pancreatitis].

The experiments were performed on 34 dogs. The subcutaneous injection of boiled pancreatic juice was established to increase the survival of dogs in acute destructive pancreatitis (p < 0.001) and to decrease the specific volume damage to acinar (p < 0.01) and interstitial (p < 0.05) pancreatic tissue. It was also shown that boiled pancreatic autojuice exerted the therapeutic effect in chronic pancreatitis complicated by pancreatic fistulas. Five peptide components with a molecular mass from 4168 D to 1000 were discovered in the canine boiled pancreatic juice by liquid high pressure chromatography. Fractions with a molecular mass between 2187 and 1348 D were found to correspond to biologically active peptides, in particular to somatostatin. It is assumed that the therapeutic effect of boiled pancreatic juice used in acute pancreatitis may be due to somatostatin. Apparently pancreatic juice may turn an efficient therapeutic agent in acute pancreatitis and pancreatic fistulas.


[The ajoene blockade of integrin-dependent processes in an HIV-infected cell system].

Ajoene, (E,Z)-4,5,9-trithiadodeca-1,6,11-triene-9-oxide, isolated from extracts of garlic (Allium sativum) has been previously shown to inhibit platelet aggregation by inactivating allosterically the platelet integrin, GP IIb/IIIa. The structural and functional similarity of integrins led the authors to suggest that ajoene may also inhibit adhesive interactions and fusion of leukocytes. Synthetic stereoisomers of ajoene synthesized by the authors exhibited equal antiaggregatory activities (IC100 approximately 50 microM for platelets; IC100 approximately 10 microM for fMLP-stimulated neutrophils). Racemic ajoene inhibited the fusion of H9 cells with HIV-infected H9:RF cells (IC50 approximately 45 microM; 16 h of incubation) and also exhibited a degree of antiviral activity (IC50 approximately 5 microM as assessed by inhibition of HIV-1/CEM/Lav 1 Bru replication in CEM13 cells; m. o. i. 0.1; 72 h). A considerable increase in the latter became evident when the compound was administered in aliquots of 50 microM per 12 h of incubation (inhibition by 30%; total concentration 0.25 microM; 72 h).(ABSTRACT TRUNCATED AT 250 WORDS)


Coupling among changes in energy metabolism, acid-base homeostasis, and ion fluxes in ischemia.

This article attempts correlating changes in cellular energy metabolism, acid-base alterations, and ion homeostasis in ischemia and other conditions. It is emphasized that loss of ion homeostasis, with thermodynamically downhill fluxes of K+, Ca2+, Na+, Cl-, and H+, occurs because energy production fails and (or) ion conductances are increased. In ischemia, energy failure is the leading event but, in hypoglycemia, activation of ion conductances is what precipitates energy failure. The initial event is a rise in K+ e, at least in part caused by activation of K+ conductances modulated by Ca2+ or ATP/ADP ratio. Secondarily, this leads to release of excitatory amino acids and massive activation of unspecific cation (and anion) conductances. Production of H+ occurs in states characterized by energy failure (ischemia and hypoxia) or by alkalosis (hypocapnia and ammonia accumulation). H+ equilibrates between intra- and extra-cellular fluid via nonionic diffusion of lactic acid, and transmembrane fluxes of H+ or HCO3- via ion channels. Since the relationship between lactate and either pHi or pHe is linear, there are no abrupt pH shifts explaining why hyperglycemia worsens ischemic damage. The reversible insults seem to induce a sustained stimulation of H+ extrusion from cells giving rise to intracellular alkalosis and extracellular acidosis.


Astrocytes, as well as neurons, express a diversity of ion channels.

The electrophysiologist's view of brain astrocytes has changed markedly in recent years. In the past astrocytes were viewed as passive, K+ selective cells, but it is now evident that they are capable of expressing voltage- and ligand-activated channels previously thought to be restricted to neurons. The functional importance of most of these ion channels is not understood at present. However, from studies of astrocytes cultured from different species and brain regions, we learned that like their neuronal counterparts astrocytes are a heterogeneous group of brain cells showing similar heterogeneity in their ion-channel expression. Not only are subpopulations of astrocytes within areas of the brain equipped with specific sets of ion channels but, furthermore, regional heterogeneity is apparent. In addition, astrocyte ion channel expression is dynamic and changes during development. Some ion channels are only expressed postnatally, yet others appear to be expressed only during certain stages of development. Interestingly, the expression of some astrocyte channels, including Na+, Ca2+, and some K+ channels, appears to be controlled by neurons via mechanisms that are presently unknown. Some studies suggest roles for astrocyte channels in basic cell processes such as cell proliferation. Thus, although the role of some astrocyte channels remains unclear, our understanding of astrocyte physiology is starting to take shape and points towards roles of ion channels not involved in electrogenesis.


Aminergic and peptidergic elements and actions in a cardiac parasympathetic ganglion.

Correlated histochemical, immunocytochemical, and electrophysiological experiments have been undertaken to identify putative neurotransmitter-neuromodulator substances in cells and fibers in the parasympathetic cardiac ganglion of the mudpuppy, Necturus maculosus, and to determine the action of these agents on the properties of the parasympathetic postganglionic neurons. The mudpuppy cardiac ganglion contains two neuron types: large parasympathetic postganglionic neurons and smaller intrinsic neurons initially identified as small intensely fluorescent cells. We have shown that the postganglionic neurons contain both acetylcholine and a galanin-like neuropeptide. Also, we have demonstrated that the intrinsic neurons contain a number of different biogenic amines such as dopamine and serotonin, as well as neuropeptides including a substance P-like peptide and a galanin-like peptide. The results of these studies indicate that the anatomical and histochemical organization of the mudpuppy cardiac ganglion is more complex than that seen in other amphibians and is very similar to that found in most mammalian species. Previously, we showed that galanin has actions that make it of interest as a potential inhibitory neurotransmitter in the mudpuppy cardiac ganglion. Galanin hyperpolarizes and decreases membrane excitability in most parasympathetic neurons. Here we show that galanin initiates membrane hyperpolarization by activating a voltage- and time-dependent potassium conductance. We also present the initial results of ongoing studies which indicate that calcitonin gene-related peptide can depolarize some of the parasympathetic neurons as well as evidence that serotonin initiates depolarization in many parasympathetic neurons. This serotonin-induced depolarization consists of an initial transient depolarization followed by a longer, more slowly developing depolarization. Action potential activity is stimulated during the initial period of depolarization, but depressed during the later, slow depolarization. The results of these electrophysiological experiments suggest that many of the bioactive substances that have been identified in the different cells and nerve fibers within the cardiac ganglion affect the excitability of the postganglionic neurons. In conclusion, we suggest that the results of the studies summarized in this review demonstrate that the cardiac ganglion in the mudpuppy is not simply a relay station. Rather, the cardiac ganglion has a complex organization and exhibits a diversity of physiological responses, indicating that it very likely is another site of integration for control of cardiac function.


Molecular mechanisms of open-channel blockade in nicotinic acetylcholine receptors of autonomic ganglia neurons.

The results of recent attempts to estimate the dimensions of ionic channels in nicotinic acetylcholine receptors of sympathetic and enteric ganglia neurons are reviewed. The channel dimensions, obtained from comparison of the sizes of the open-channel blocking molecules with their blocking activities, are 6.1 x 8.3 A (1 A = 0.1 nm) in both sympathetic and enteric ganglia. None of the competitive ganglionic blockers fit within this channel size. In addition, a chemical structure that binds the open-channel blockers in ganglionic nicotinic acetylcholine receptors is suggested to be formed by serine and threonine residues, as found by comparing the differences between the structures of the neuronal and muscle nicotinic acetylcholine receptors with the differences in their pharmacology.


Inhibition of neutrophil priming and tyrosyl phosphorylation by cepharanthine, a nonsteroidal antiinflammatory drug.

Receptor-mediated superoxide (O2.-)-generation and tyrosyl phosphorylation of neutrophil proteins, such as 58, 65, 84, 108 and 115 kDa, were enhanced by priming cells with granulocyte colony stimulating factor (G-CSF) [Akimura, K. et al. Arch. Biochem. Biophys. 298: 703-709, 1992]. To elucidate the possible involvement of tyrosyl phosphorylation of neutrophil proteins in the enhancing mechanism of O2.- generation, the effect of cepharanthine, a biscoclaurine alkaloid that inhibits phorbol 12-myristate 13-acetate (PMA)- and receptor-mediated O2.- generation, on the priming of human peripheral neutrophils (HPPMN) was studied. Both enhancement of formyl-methionyl-leucyl- phenylalanine (FMLP)-mediated O2.- generation and tyrosyl phosphorylation of some neutrophil proteins, i.e., 115, 108 and 84 kDa proteins, by HHPMN after treatment with G-CSF were strongly inhibited by cepharanthine in a concentration- and treatment-time-dependent manner. In contrast, inhibition of PMA-mediated O2.- generation by cepharanthine was weak and independent of treatment time. These results suggest that cepharanthine might inhibit the priming step of neutrophil activation concomitantly with its inhibition of the tyrosyl phosphorylation of some neutrophil proteins that might underlie the mechanism for priming of neutrophils with G-CSF.


Myelin associated glycoproteins confer heterophilic adhesion properties to an immortalized optic nerve derived cell line.

We have transfected an immortalized optic nerve-derived cell line with the cDNA's encoding the two isoforms of MAG. Our aim was to assess whether expression of L-MAG (72 KDa) and S-MAG (67 KDa) in these cells confer adhesion properties when a suspension of single cells is allowed to aggregate. The selected cell lines expressed MAG mRNAs and proteins of the appropriate molecular size, and the proteins were targeted correctly to the plasma membrane. Both L-MAG and S-MAG-expressing transfectants exhibited enhanced self-adhesive properties, aggregating with faster kinetics and forming larger aggregates than MAG-negative control cells. The interaction appears to be mostly heterophilic since MAG+ and MAG- cells which were labeled with a fluorescent probe bound equally well to pre-aggregated MAG+ transfectants and their interaction was blocked by monoclonal anti-MAG antibodies. A further finding which supports the role of MAG in adhesion was the observation that MAG was preferentially localized at the junctions between cells, in confluent cultures.


Diagnostic value of analysis of cystatin C and protein HC in biological fluids.

Cystatin C, a 13 kDa-protein, is produced by most nucleated cells and is catabolized by the renal tubular cells after passing the glomerular filter. It belongs to the family 2 of the cystatin superfamily of proteins. The function of cystatin C is to regulate the activity of cysteine proteinases and cystatin C seems to be the main cysteine proteinase inhibitor of most investigated human biological fluids. Its normal level in plasma is 0.8-2.5 mg/l, in cerebrospinal fluid 4-14 mg/l and in urine 0.03-0.3 mg/l. The production rate of cystatin C is remarkably constant and its plasma concentration can therefore be used as a reliable measure of the glomerular filtration rate (GFR). Indeed, the cystatin C plasma concentration is more closely correlated to the GFR than the plasma levels of creatinine and all other investigated low molecular weight proteins, including beta 2-microglobulin and retinol binding protein. Protein HC, alias alpha 1-microglobulin, is produced by the liver as a 27 kDa-glycoprotein. It belongs to the lipocalin superfamily of hydrophobic ligand binding proteins and more than 50% of the normal plasma amount of protein HC is present as a high molecular weight HC-IgA complex carrying antibody activity. The plasma concentration of free protein HC is, in contrast to that of HC-IgA, mainly determined by the GFR. The normal values for the plasma concentrations of HC-IgA and free protein HC are 36-620 mg/l and 14-26 mg/l, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Utility of filtration markers to monitor the quality of glomerular function.

Diabetes mellitus can lead, along the years of its course, to chronic renal failure in a high proportion of cases. An early risk-indicator of later diabetic nephropathy is the presence of microalbuminuria, but it usually takes about fifteen to twenty years to appear. Before that, no clinical signs can disclose the underlying alterations of glomerular basement membrane that will eventually bring forth overt nephropathy. The usefulness of the altered excretion of isoenzymes of amylase as an early marker of the glomerular charge selectivity was tested in 202 juvenile onset insulin-dependent diabetics, compared with 51 normal subjects matched for age and sex. The diabetic patients studied showed increased excretion of salivary amylase into urine. The salivary to pancreatic amylase ratio of concentrations in urine was always below 1 in normal subjects, and was increased over 1 in 33.2% of diabetics, although microalbuminuria was present only in 26.2% of patients. The excretion of other proteins was within reference values in the majority of cases, indicating that the kidney was not seriously affected in those patients. Moreover, the altered salivary to pancreatic amylase ratio in urine was more prevalent than microalbuminuria (36.6% vs 18%) in the first decade of the evolution of the diabetes. These results indicate that the ratio of excretions of both isoamylases into urine is a more sensible and earlier marker of altered glomerular charge barrier for anionic proteins.


Spatially restricted expression of fibroblast growth factor receptor-2 during Xenopus development.

The fibroblast growth factors (FGFs) play a role in Xenopus laevis embryonic development, particularly in the induction of ventral-type mesoderm. We have isolated a full-length cDNA from Xenopus that we have designated Xenopus fibroblast growth factor receptor-2 (XFGFR-2), with significant amino acid sequence similarity to the previously described bek gene (FGFR-2). We expressed the XFGFR-2 cDNA in COS1 cells and showed that it functions as an FGF receptor by binding radiolabeled FGF-2. RNA gel blot analysis demonstrates that unlike Xenopus fibroblast growth factor receptor-1 (XFGFR-1), XFGFR-2 mRNA expression begins during gastrulation and continues through early tadpole stages. Whole-mount in situ hybridization demonstrates that XFGFR-2 mRNA is localized to the anterior neural plate in early neurula stage embryos. Later in development, XFGFR-2 expression is found in the eye anlagen, midbrain-hindbrain boundary and the otic vesicle. In addition, XFGFR-2 transcripts are expressed in animal caps in a manner that is independent of mesoderm-inducing factors. These results indicate that XFGFR-2 may have a role in development that is distinct from that of XFGFR-1.


Timing of neuronal death following successive blockade of protein synthesis and axoplasmic transport in the axonal target territory.

Neurons in the isthmo-optic nucleus (ION) of chick embryos can withstand a substantial cycloheximide-induced reduction in protein synthesis in their target territory, the retina, at the very time when their survival is known to depend on a retrograde signal from the latter. We here test the hypothesis that this resistance to the cycloheximide injection might be due to the accumulation in the ION of a reserve of retina-derived trophic molecules (or of resulting second messengers). Following an intraocular injection of cycloheximide at E15 to deplete the hypothetical reserve in one ION, both eyes received injections of colchicine (which blocks axoplasmic transport) at E16. The resulting time course of cell death was very similar in the two IONs, which refutes the hypothesis. The most plausible alternative is that there is a reserve of trophic substance in the retina capable of maintaining the ION for about 1 day.


Endogenous and exogenous alpha-fetoproteins as differential markers of cultured neonatal mouse Schwann cells and fibroblasts.

alpha-Fetoprotein (AFP) and AFP-gene transcripts were demonstrated in vitro in Schwann (S) cell and fibroblast (F) cell cultures of neonatal mouse origin. All S and F cells of primary cultures and of established cell lines expressed the AFP gene. AFP mRNA was detected by an in situ hybridization technique using a 35S-AFP-cDNA probe. AFP was localized by immunocytoperoxidase labelling using purified anti-AFP antibodies. The amounts of stained endogenous AFP, estimated semiquantitatively, were about 3-fold higher in S cells than in F cells. After incubating the cultures with exogenous mouse AFP, both S and F cells showed significant ability to take up the protein; the amount of internalized protein was found to be higher in F cells than in S cells. Moreover, the uptake of AFP fluorescein conjugates (FITC-AFP), estimated quantitatively by fluorometry, also gave higher values for F cells. The cytoplasm of F cells exhibited a characteristic fluorescence pattern, strongly illuminated and dispersed grains; the cytoplasm of S cells was regularly labelled. If exogenous FITC-AFP uptake could be used to distinguish labelled F cells from S cells (with application for identification and selection of F cells), the immunocytochemically stained endogenous AFP could allow S cells to be distinguished from F cells (using the dilutions of antibodies still staining the S cells but which lead to the absence of F-cell labelling). The two procedures, which can be used independently or together, may constitute differential markers for S cell and F cultures in, i.e., nerve regeneration of neurofibroma studies using the model of mixed S and F culture also containing other types of cells.


Ultrastructural localization of basic lysine-rich proteins during the nucleologenesis in preimplantation bovine embryos.

The process of nucleologenesis was examined in preimplantation bovine embryos by using ethanolic phosphotungstic acid (E-PTA) at the electron microscopic level. E-PTA binds basic lysine-rich proteins and thus makes possible detection of their localization and distribution. In blastomere nuclei of 2-, 4- and early 8-cell embryos, only nucleolus precursor body/bodies (NPBs) appear, being formed from a mass which is homogeneously stained with E-PTA. In cow embryos, the whole nucleologenesis is situated in the 8-cell stage. During the following step of nucleologenesis, the NPB with a big central area (named NPB vacuole) is formed. Fibrillar mass around the central vacuole is intensively stained, especially in the regions in close vicinity to the central vacuole. Many clumps of E-PTA-positive, regularly dispersed material are found inside the vacuole. The next step of nucleologenesis is characterized by the presence of NPB with secondary vacuoles that are also filled with clumps of E-PTA-positive material. Small, intensively stained areas are visible at sites that are probably identical with those where a dense fibrillar component is formed. Until the end of the 8-cell stage, as well as in the morula and early blastocyst, typical fibrillogranular nucleoli are present. These nucleoli have 3 basic components--fibrillar centres (FC), dense fibrillar components (DFC) and granular components (GC) which stain with different increasing intensity. FC and DFC show a strong (particular) reaction while the GC are stained to a lesser degree. In all examined stages of embryonal development, the E-PTA positivity was found within the perinucleolar chromatin and on the clumps of heterochromatin. An analogical localization of basic and acidic regulatory proteins in the examined developmental stages is discussed.


A novel panel of monoclonal antibodies against beta-galactosidase of Escherichia coli and its versatility for detection of recombinant expression products.

A panel of seven mouse monoclonal antibodies (BG-O1 - BG-07) raised against beta-galactosidase (beta-gal) from E. coli was characterized in respect of their binding to beta-gal and to fusion proteins. The antibodies were beta-gal specific, recognized six different antigenic determinants on beta-gal molecule and some of them inhibited catalytical activity of the enzyme. The antibodies reacted with C-terminal beta-gal fusion proteins in the native state as well as after Western blotting. BG-02 antibody was successfully used for immunofluorescence detection of cells transfected with vectors containing the lacZ gene.


[Changes in the serological indices of the immune status of workers in contact with raw foodstuffs contaminated by mycotoxins].

The immune status was estimated in workers engaged into the grain processing and exposed to constant hazards of the industrial environment (contact with the goods contaminated by mycotoxins). Studies revealed in workers the certain changes of serum Ig, complement fractions, proteins of the acute phase of inflammation. The importance of immunologic changes as criteria for adequate prophylactic measures was stressed.


Growth in malnutrition related to gastrointestinal diseases: coeliac disease.

Coeliac disease in children is frequently associated with a slow growth rate. This observation may be linked to the malabsorption that occurs in these patients; however, the underlying mechanism remains unknown. To better understand this phenomenon, we have studied the growth patterns of 153 patients with coeliac disease for 2-9 years. Gastro-intestinal biopsies were performed before and after gluten exclusion. In a second group of 79 children, somatostatin levels and binding properties in the plasma and jejunal mucosa were measured. In a third group of 40 patients we measured insulin-like growth factor I (IGF-I) and IGF-binding protein 3 (IGF-BP3) levels. We found that in children diagnosed before 2 years of age weight was the most affected growth parameter. In children diagnosed after this age, height was more affected. Suppression of gluten intake induced an acceleration of growth velocity. Although plasma levels of somatostatin were not significantly altered, somatostatin concentrations in the jejunal mucosa of patients in the active phase of the disease were significantly elevated (p < 0.05). Children with coeliac disease exhibited significantly lower levels of IGF-BP3 when compared to patients with normal stature and growth velocities. In contrast, these patients showed an increase in IGF-BP3 levels after gluten exclusion from the diet.


Immunofluorescence detection of the vimentin epitope in chromatin structures of cell nuclei and chromosomes.

We describe immunofluorescence detection of the vimentin epitope, recognized by monoclonal antibody VI-01, in chromatin structures of eukaryotic cell nuclei and chromosomes. The approach used is based on increased sensitivity of 5-bromodeoxyuridine (BrdU)-substituted DNA to UV irradiation-induced crosslinking of DNA with proteins in vivo, by which the proteins interacting with chromosomal DNA can be immunovisualized in situ.


Differential expression of insulin-like growth factors I and II (IGF I and II), mRNA, peptide and binding protein 1 during mouse palate development: comparison with TGF beta peptide distribution.

Development of the mammalian secondary palate involves a series of epithelial mesenchymal interactions: during one of these, a mesenchymal signal specifies regionally distinct palatal epithelial differentiation. Extracellular matrix molecules and soluble growth factors may be involved in this signalling process. In this study, we have mapped the expression of the genes for insulin-like growth factors (IGF I and II), the peptides they encode, and the IGF binding protein 1 (IGF BP-1) during murine palatogenesis (embryonic days (E) 12-15). IGF-I gene expression was below detectable levels in the craniofacial region at all ages. IGF-I peptide was at the threshold of immunocytochemical detection and widely distributed in the palatal mesenchyme, decreasing in staining intensity from E12 to E14. By contrast, IGF-II mRNA was intensely localised in several tissues. IGF-II gene expression within the forming palate was developmentally regulated. In the vertical palatal shelves (E12 to E13) IGF-II gene expression was absent. On early E14, in the horizontal prefusion palate, significant expression was present in the palatal mesenchyme, but not the epithelium. Once palatal fusion had occurred, mesenchymal expression fell rapidly to undetectable levels. IGF-II mRNA was next detectable in the secondary palate on late E15 at sites of membranous bone formation. By contrast to the mRNA distribution, IGF-II peptide was localised predominantly in the palatal epithelia (particularly the nasal and medial edge epithelia) but also in the mesenchyme of the E14 prefusion palate. Significantly, the IGF binding protein had a similar distribution pattern to the IGF-II peptide. At all ages, the developing tongue myotubes labelled heavily for IGF-II mRNA, protein and binding protein. These data suggest that IGF-II may play a localised paracrine role during murine palatogenesis, perhaps in the mesenchymal signalling of epithelial differentiation. IGF-II may also serve to coordinate the development of the tongue and palate. The distribution of IGF-II peptide was very similar to that of TGF-beta, suggesting a possible interactive role of these growth factors during palate development. Finally, evidence that the IGF-II gene is imprinted (Ferguson-Smith et al. 1991) and may be the target for uniparental disomy in the human Beckwith Wiedemann syndrome (Henry et al. 1991), which is characterised by the overgrowth of tissues (especially the tongue) expressing IGF II in the embryo, indicates the necessity of reanalysing human cleft palate families for disruption (including uniparental disomy) of the genes encoding IGFs, their receptors and binding proteins.


The primary structure of porcine aminoacylase 1 deduced from cDNA sequence.

A cDNA encoding the complete amino acid sequence of aminoacylase 1 (N-acylamino acid aminohydrolase, ACY-1) [EC 3.5.1.14], a dimeric metalloprotein having two Zn2+ in the molecule, which catalyzes the deacylation of N-acylated L-amino acids except L-aspartic acid, has been isolated from porcine kidney lambda gt10 cDNA library and sequenced. From sequence analysis of the cDNA and the N- and C-terminal amino acid analyses of the purified protein, it is deduced that porcine kidney ACY-1 consists of two identical subunits (M(r) 45,260), each of which consists of a single chain of 406 amino acids with acetylalanine at the N-terminus. A cDNA encoding porcine liver ACY-1 was also cloned. The amino acid sequence deduced from the nucleotide sequence of the cDNA from porcine liver was identical to that deduced for porcine kidney ACY-1. Northern blot analysis suggested that ACY-1 is more highly expressed in kidney than in liver. Comparison of the amino acid sequence of porcine ACY-1 with those of other Zn2+-binding metalloenzymes showed no significant homologies in either the overall sequence or the consensus sequences for the metal binding sites. This indicates that ACY-1 is a new type of metalloprotein.


Isolation of cDNA for bovine stomach 155 kDa protein exhibiting myosin light chain kinase activity.

Two proteins with myosin light chain kinase activity and electrophoretic molecular weights of 155,000 and 130,000 were each isolated from bovine stomach smooth muscle [Kuwayama, H., Suzuki, M., Koga, R., & Ebashi, S. (1988) J. Biochem. 104, 862-866]. The 155 kDa component showed a much higher superprecipitation-inducing activity than the 130 kDa component, when compared on the basis of equivalent myosin light chain kinase activity. In this study, we isolated a cDNA for the entire coding region of the 155 kDa protein. The deduced amino acid sequence revealed a high degree of similarity to those of chicken and rabbit smooth muscle myosin light chain kinases. Multiple motifs, such as three repeats of an immunoglobulin C2-like domain, a fibronectin type III domain, and unusual 20 repeats of 12 amino acids were detected in the sequence. Part of the amino-terminal sequence was similar to that of the actin- and calmodulin-binding domain of smooth muscle caldesmon. These observations suggest that the 155 kDa protein has additional functions other than its enzymatic activity. Two mRNAs of 6.0 and 2.6 kb in length in the bovine stomach smooth muscle RNAs were hybridized with cDNA probes. The 2.6-kb RNA probably encodes telokin, which is the carboxyl terminus of smooth muscle myosin light chain kinase. mRNAs with identical lengths were also detected in bovine aorta.


Molecular cloning and partial characterization of a novel collagen chain, alpha 1(XVI), consisting of repetitive collagenous domains and cysteine-containing non-collagenous segments.

In an effort to identify new members of the collagen family, we screened a human placenta cDNA library with a collagenous probe. A novel 3.7 kb cDNA was identified encoding an open reading frame of 1,186 amino acids and containing a termination codon. The predicted polypeptide consists of 9 repetitive collagenous (stretches of Gly-X-Y) and several non-collagenous segments. Two cysteinyl residues separated by two amino acid residues (Cys-X-X-Cys) are regularly located in the N-terminal region of each non-collagenous segment. The deduced amino acid sequence described above is distinct from those of known types of collagen. Therefore, this novel collagen chain is designated alpha 1(XVI). Northern blot analysis revealed an alpha 1(XVI) mRNA of 5.2 kb, indicating that the overlapping cDNA clones isolated in this study covered nearly three-fourths of the mRNA. As a tool for further study on the expression of type XVI collagen, we prepared an antibody against the nonadecapeptide CFLSLERPRAEEARGDNSE, derived from the putative translation product of the cDNA. In immunoblot analysis, the antibody recognized a 160 kDa protein, which was bacterial collagenase-sensitive. Immunohistochemical stainings of human placental tissues with anti-peptide antibody revealed a positive reaction with amnion, the membranous tissue lining the amniotic cavity. The gene of alpha 1(XVI) chain, COL16A1, is mapped on the short arm of human chromosome 1 (1p13-p34).