Some pharmacological actions of aloe extracts and Cassia abbreviata on rats and mice. The effects of crude extracts (500 mg/kg IP and 500-1000 mg/kg PO) of three species of aloes and Cassia abbreviata in rats and mice were studied, particularly as regards their abortifacient actions. The LD50 values were as follows: A. globuligemma < 250 mg/kg IP; A.chabaudii 250-500 mg/kg IP; A. cryptopoda > 1500 mg/kg IP and C.abbreviata 500-750 mg/kg IP. Thus the most toxic was A.globuligemma. Their most visibly striking toxic effects in rats were CNS depression, and post mortem investigations showed widespread haemorrhagic lesions. Administration of the aloes and A.abbreviata to pregnant mice and rats did not cause expulsion or resorption of the foetuses. Several rats died within 36 hours of injection of A.chabaudii and A.globuligemma, but even in these rats there was no expulsion or resorption of the foetuses. Rats which survived the treatment delivered normal sized, healthy litters at term. It is suggested that neither the aloe species test nor C.abbreviata possess abortifacient activity at doses which are not toxic to the animals.


Correlations between weights of newborn babies, placental parameters and gestational age. The relationships between foetal weights, placental parameters and gestational ages were studied in 200 placentae. In addition to confirming that foetal weight, placental weight and gestational age correlated, the results showed that baby weight, placental weight, placental surface area correlate to the length of cord and that the length of cord correlates to gestational age (p < 0.05). These correlations are reported for the first time and are difficult to explain. It is recommended to establish birthweight standards in order to assess intra-uterine growth retardation, and to re-examine the correlations reported here through prospective studies as they might be relevant clinically and provide important clues on morbidity prospects of an individual during postnatal life.


Long-term results of tricuspid annuloplasty according to DeVega. The long-term results on 36 patients undergoing tricuspid annuloplasty according to DeVega over the years 1981-1990 were analyzed. All had mitral valve replacement or repair in addition to the above procedure. Moreover, three patients had aortic valve replacement while aortocoronary bypass and interatrial septal defect closure was performed in two cases each. Duration of follow-up ranged between 6 to 72 months (mean 36 months). All patients were examined by echocardiography. Significant tricuspid regurgitation was found in three of the patients operated on, manifest clinical signs of the condition were present in one female patient only. Functional evaluation reassigned 72% of the operated patients from Class III and IV to Class I and II, and apparent subjective improvement was reported by 81% of patients. The analysis showed that indication of patients for simultaneous management of tricuspid regurgitation associated with mitral valve disease is justified, and that the surgical technique employed is appropriate.


Carotid artery involvement in patients with ischaemic heart disease prior to myocardial revascularization. A carotid arterial bed with sclerotic lesions in patients with ischaemic heart disease before myocardial revascularization in extracorporeal circulation raises the risk of development of serious irreversible dysfunction of the central nervous system. The authors report on the results of ultrasound examination of the carotid arteries in 284 patients, hospitalized at the Department of Cardiac Surgery of University Hospital at Hradec Králové prior to myocardial revascularization between 1986-1990. Based on their results, and in agreement with data of the literature, the authors recommend to perform non-invasive examination of the carotid arterial bed in all patients prior to myocardial revascularization and, based on the results of the examination, to follow a strictly individualized investigational and therapeutic algorithm.


A comparison of the protective effect of a modified StTH solution and HTK-B on the energy and functional status of the isolated rat heart. Using a model of the isolated beating rat heart, the authors compared the protective effect of St. Thomas Hospital cardioplegic solution enriched with glucose and mannitol (StTH-M) and Bretschneider solution (HTK-B). Results showed that, during 120-minute global ischaemia in cardioplegia, StTH-M was able to maintain levels of high-energy phosphates comparable with those found in a group of hearts perfused with HTK-B at 20 degrees C only when the temperature had been decreased to 12-15 degrees C. Under these conditions, repair of metabolic and functional parameters during post-ischaemic perfusion was also similar in both groups.


Urinary albumin excretion in patients with renovascular hypertension. Twenty-four hour urinary excretion of albumin (UEalb), IgG and beta-2 microglobulin was investigated at a 3 hour-interval in a control group (C) of healthy subjects, in 30 patients with renovascular hypertension (RVH), and in 16 patients with essential hypertension (EH). Mean UEalb in RVH was significantly higher than in C. A significant direct correlation was demonstrated between diastolic blood pressure and UEalb (p < 0.01). Microalbuminuria (MA) > or = 30 micrograms.min-1 was found in about 18% of RVH patients; it was higher than 16.7 micrograms.min-1 in approx. 31%. These results did not substantially differ from those obtained in patients with EH. The cause for increased UEalb in hypertensive patients may be functional, haemodynamic changes, or structural ones. In either case, MA indicates renal injury, and these patients should be given increased attention when monitoring their blood pressure and when selecting antihypertensive drugs.


Follow-up of outpatients with essential hypertension. A comparison of three methods of blood pressure measurement. Compared with conventional, routine blood pressure measurement in the outpatient department of a hospital, 24-hour blood pressure monitoring provides more comprehensive information about blood pressure values in day-to-day life of the hypertensive patient. The hospital setting with all its stimuli affects the hypertensive patients so that their values will be statistically significantly higher compared with normal ones. Recent years have seen the introduction into clinical practice of self-monitoring by the patients themselves. Comparison of these three methods for blood pressure measurements revealed that self-monitoring may provide fairly accurate values comparable with those obtained by the "standard method", i.e., 24-hour monitoring. In view of the low variability of the mean value of blood pressure over a period of several days, self-monitoring can be employed in groups of selected outpatients on long-term follow-up.


Metabolic effects of enalapril in the treatment of essential hypertension. In an open two-month study with an initial placebo period, the effect of enalapril on glucose tolerance, insulin (IRI) sensitivity and lipid profile was evaluated in 20 patients with mild to moderate essential hypertension. The following results were obtained: 1. Enalapril produced a favourable effect of blood pressure both in monotherapy and if combined with a diuretic. 2. Therapy did not lead to significant differences in blood glucose, IRI or IRI/glucose increase at 1 or 2 hours of oral glucose tolerance test either in patients with monotherapy or combination therapy, and with normal or disturbed glucose tolerance, respectively. 3. Serum lipids (total and HDL-cholesterol and triglycerides) did not change significantly in any group of patients.


Prognostic value of body mass index in the development of cardiovascular diseases among Kaunas male population. The paper presents the results of a primary medical examination and of a prospective, on the average 14-year follow-up study of the male population of Kaunas aged 45-59 years (n = 2452). The prevalence of overweight, according to the WHO criteria, was 48.7%, that of obesity 20.5%. With increasing body mass index (BMI) there occurred a statistically significant increase of main risk factors of ischaemic heart disease (IHD) and of IHD itself. The results of the prospective study showed a U-shaped relationship between BMI and mortality from all cardiovascular diseases, and a J-shaped relationship between BMI and mortality from IHD and morbidity of acute myocardial infarction, respectively. The same relationship held true even after standardization of the level of all other risk factors using the multiple logistic function. Thus, the overweight was confirmed as an independent IHD risk factor in the given population.


A long history of the left main coronary artery occlusion. The case of a 70-year-old woman with severe angina pectoris for 12 years and occlusion of the left main coronary artery is presented. The reasons for this long history seemed to be adequate collateral circulation, hibernating myocardium, as well as incredible patience of the patient.


Circulating endothelial cells in surgical patients. The circulating endothelial cells (CEC) in healthy volunteers and surgical patients before and after operations were counted. The results showed that CEC counts of the patients were much higher than those of the volunteers. After operation, however, patients' CEC counts returned to control levels. It is suggested that detection of CEC may be useful for quantitatively evaluation of the injury to blood vessels in surgical patients. The operation may not only remove the source of injury, it also eliminates the injured microvascular bed which may be important to maintain homeostasis. Another possible interpretation of the results may be related to time lag and mental stress.


Age-related changes in the levels of IgG, IgM and IgA antibodies to type I collagen in the sera of normal human subjects. Serum samples from healthy subjects of different ages (within the age range of 1-75 years) were tested for the presence of anti-type I collagen (anti-CI) IgG, IgM and IgA by enzyme-linked immunosorbent assay (ELISA). All the tested sera showed anti-CI antibodies from the three immunoglobulin classes with the following age-related regularities: 1. Anti-CI IgG and IgM showed statistically non-significant changes up to the age of 60 decreasing thereafter. 2. Anti-Ci IgA changed negligibly up to the age of 40 increasing with age thereafter. The established age-related changes in the levels of anti-CI antibodies may serve as a basis for future studies of normal and pathological turnover of type I collagen.


Pulse repetition rate and duration affect the responses of bat auditory cortical neurons. Under free field stimulation conditions, we studied the effect of pulse repetition rate and duration on the responses of auditory cortical neurons of the big brown bat, Eptesicus fuscus. The best frequency (BF) and minimum threshold (MT) of each recorded neuron were first determined with a 4 ms pulse delivered from a specific point of the bat's frontal auditory space at which the neuron had maximal spatial sensitivity. Then a 4 ms pulse was delivered repetitively at different rates with an intensity in 10 dB increments above the neuron's MT and its number of impulses and response latency to each combination of repetition rate and intensity were recorded. From the series of isorepetition rate-intensity functions, we identified the neuron's best intensity (BI) which elicited a maximal number of impulses. By using a 4 ms BF and BI pulse delivered at a wide range of repetition rates, we then determined the neuron's best repetition rate (BRR) to which the neuron discharged maximally. The effect of pulse duration on the neuron's responses was subsequently studied by recording its number of impulses and latency to a BF, BI and BRR pulse delivered at a wide range of durations. A neuron's best duration (BD) to which the neuron discharged maximally was also identified. We found that cortical neurons sequentially isolated with an orthogonally penetrated electrode have comparable BFs, MTs and latencies. In addition, high BF neurons tend to have higher MTs and shorter latencies than low BF neurons. Most intensity-rate functions of cortical neurons were nonmonotonic but they can be affected by pulse repetition rate. Under repetitive single pulse stimulation, all cortical neurons show low pass characteristic to pulse repetition rate such that their number of impulses reduced drastically with increasing pulse repetition rate regardless of pulse intensity. The BRRs of most cortical neurons studied are between 1 and 3 pulses/s and their BDs are between 0.5 and 10 ms. Response latencies of cortical neurons generally shortened to a plateau level with increasing pulse intensity. In contrast, they increased with pulse repetition rate and varied in different manner with pulse duration. Nevertheless, a cortical neuron generally responds with a shortest latency to a pulse delivered at its BF, BI, BRR and BD. Thus in auditory signal processing, a neuron's responses to a wide combination of pulse intensity, duration and repetition rate enhance its versatility in signal detection. Its maximal response to a specific combination of signal duration, repetition rate and intensity provides a neural basis for fine analysis of signal features.


Neurofibrillary degeneration and microtubule associated protein tau. Alzheimer disease is characterized by neurofibrillary pathology containing paired helical filaments (PHF). These abnormal filaments consist of a modified form of microtubule associated protein tau. The modification involves phosphorylation. In this mini review, we summarize recent studies regarding the differences between normal tau and PHF-tau, focusing especially on the extent and the site of phosphorylation. We also discuss the mechanisms possible involved in the development of PHF.


Phonology and orthography in visual word recognition: evidence from masked non-word priming. Three lexical decision experiments in French investigated the effects of briefly presented forward-masked non-word primes on latencies to phonologically and/or orthographically related targets. At 64-msec prime presentation durations, primes that are pseudohomophones of the target produced facilitatory effects compared to orthographic controls, but these orthographically similar non-word primes did not facilitate target recognition compared to unrelated controls. These results were obtained independently of target word frequency and independently of the presence or absence of pseudohomophone targets in the experimental lists. With a 32-msec prime duration, on the other hand, pseudohomophone and orthographic primes had similar effects on target recognition, both producing facilitation relative to unrelated controls. The results are discussed in terms of the time course of phonological and orthographic code activation in the processing of pronounceable strings of letters.


Do visual images have Gestalt properties? Recent research has demonstrated that percepts and images share processing resources. A natural consequence of this evidence is to ask what kind of properties are shared by the two representational systems, i.e. images and percepts. To what degree, for example, do images share the complex organization of visual percepts? This paper investigates whether percepts and images share Gestalt properties. Four experiments were conducted to study this hypothesis. In all experiments subjects were presented with an auditory message in which approximately 15 locations in a matrix were defined by giving the co-ordinates of the cells. Half of the stimuli presented some "good" form, whereas in the other half the locations of the pieces were scattered. Subjects were systematically less able to recall the locations of the random forms. Therefore, it could be argued that the "good" forms help in image construction, even though the elements were auditorily presented. Effects of varying presentation speed, order, and of requiring a mental rotation before recall support this conclusion.


The representation of objects and their attributes in memory: evidence concerning memory for location. The general principle is advanced that different attributes of objects (e.g. shape and colour) are more readily associated when they are attributes of the same object than when they are attributes of different objects. Previous studies provide support for the principle, for they have shown that a shape is more readily associated with its own colour than, for example, with the colour of its background. In the present experiments, the principle was applied to the attributes of shape and location. In four experiments it was shown that a shape was more readily associated with its own location than with the location of another object. Differences in contiguity between the presentation of the shape and the location with which the shape was to be associated were controlled for, as were other factors. The results were interpreted as providing support for the general principle stated above. It was suggested that the general principle could be explained by supposing that memory is propositional in format.


An anxiety-related bias in semantic activation when processing threat/neutral homographs. Three experiments are reported comparing high- and low-trait anxious subjects in terms of their patterns of semantic activation in response to ambiguous primes, with one threat-related and one neutral meaning. Such primes were followed by targets related to either their threat or neutral meaning, or by unrelated targets, in a lexical decision task. Experiments 1 to 3 employed stimulus onset asynchronies (SOAs) of 750 msec, 500 msec, and 1250 msec, respectively. At 500-msec SOA all subjects showed facilitation for both meanings. At 750-msec SOA the only significant priming effect was that for the threat-related meaning in the high-anxiety group, and a similar trend was found at 1250-msec SOA. Consideration of the patterns of priming for targets following ambiguous threat/neutral primes suggest that at the longer SOAs, high-anxiety subjects consciously "lock on" to a threatening interpretation if one has been made available by earlier automatic spreading activation.


[Control of foodstuffs in international turnover. I. Documents of the European economic community]. Council Directive (89/397/EEC) is presented which harmonize official control of foodstuffs. The main principles are given, obligatory for the Member States, concerning control of foodstuffs, food additives and materials intended to come into contact with foodstuffs performed by home authorities. The proposal of supplementary Directive is discussed, dealing with problems of qualification and competence of laboratory personnel, introduction of quality assurance systems in the laboratories and the system of information exchange of foodstuffs within the Common Market.


[Problem with the presence of carcinogenic substances formed spontaneously or during technologic processing in food]. The current opinion on the spontaneously occurring or evoked by certain culinary and technological processes muta-, cancero- and teratogenic compounds are reviewed on the basis of the world literature. It was noted that these substances may form in food products of vegetal origin. Formation of carcinogenic compounds is more intensive in plants attacked by various diseases, insects or parasites. The importance of this problem should be stressed together with the need of improving knowledge in this field. The compounds under discussion are the most serious risk factor in the incidence of neoplasms in humans.


[Estimation of table salt content in daily food rations of children, youth and elderly people]. In order to estimate table salt intake in selected population groups, sodium chloride as well as sodium, potassium were determined in daily food rations. Sodium chloride was determined by Mohr's method, while sodium and potassium by flame photometry. Parallel samples of soups, bread and carbohydrates in the second courses were analysed in the same way. Samples for analysis were collected during a period of 10 days in a State Children's Home (SCH) and in Home for the Aged (HA). The table salt content in food rations from the HA was found between 14.7-24.7 g and in rations from the SCH between 7.9-19.6. In both cases the values were significantly higher than those recommended. The average table salt content in soup from the HA was 1.24 g/100 g and that from the SCH -1.65 g/100 g. A high level of salt was observed also in potatoes, groats and macaroni, the average value being 1.5 g/100g. As regards the size of the portions given, it was found, that soups as well as bread supplied 34% of the total salt content in the food rations, and the carbohydrates in the second courses brought about 14%. It was found, that about 96% of the total salt quantity in the investigated rations was derived from addition of salt during food preparation and from the processed products. The studied food rations contained relatively high quantities of potassium, that completely satisfied the demand for this element of the investigated population groups. The Na:K weight ratio was abnormal, as the results of too high sodium content.


[Determination of antibiotic residues in some food products of animal origin]. Food products of animal origin were analysed in years 1986-1990 by Sanitary-Epidemiological Stations for the presence of antibiotics residues-chloramphenicol and streptomycin. In raw milk chloramphenicol (trace) was found in 9 out 938 samples, streptomycin (3.66-19.67 ppm) in 28 out 251. In eggs chloramphenicol (trace) was present in 5 out 866 samples. In milk powder chloramphenicol was not detected.


[Evaluation of degree of freshness and value of fish and fish product t intake based on histamine and trimethylamine analysis]. The results of histamine and trimethylamine determination in three fish species (cod, herring, scomber) of various freshness are presented. In fresh fish the histamine contents were s follow: cod 0.00; scomber 1.00; and herring 0.50 mg/100 g. During storage time at room temperature (+18 degrees C), the value of the freshness indicators increased after 24 and 48 hours of storage. The highest increase was observed in scomber (21.30 and 34.00 mg/100 g), the lowest in cod (2.00 and 4.00 mg/100g). The trimethylamine value increased in different order than the histamine in the fish species. The highest TMA amount was observed in cod flesh--the lowest in scomber. According to these findings the freshness of some ready fish products was determined. The histamine level never exceeded 6 mg/100 g, always being below the "safety level" (= 10 mg/100g). The value of TMA never exceeded the limits considered for fresh fish products--15 mg N per 100 mg sample.


[Effect of the fermentation process on levels of nitrates and nitrites in selected vegetables]. The aim of this study was to follow the changes in the levels of nitrates and nitrites throughout the process of fermentation of sauerkraut from white and red cabbage and red beets. The nitrate and nitrite levels were determined in raw and fermentation as well as in red beets and "beet acid" after a week of souring. Nitrate were determined by the brucine method, while nitrates by the Griess colorimetric method. Mean reduction of nitrates in sauerkraut (in relation to raw cabbage) was ca. 55.5% and that of nitrites ca. 76.7%. In the red sour cabbage a decrease in the level of nitrates in relation to the product by ca. 84.1% and in that of nitrites by ca. 67.4% was found. The stabilization of both nitrate levels in both kinds of cabbages followed as after the second month of storage, as confirmed by statistical analysis of the results. In the red beets after the process of fermentation a decrease in the level of nitrates by ca. 91.6% was noted. An undefined portion of the studied compounds passed into the liquid. The "beet acid" contained 595.9 mg/dm1 of nitrates and 3.26 mg/dm3 of nitrites.


[Nitrate water pollution of drinking water in the Bydgoszcz district]. The study was undertaken to evaluate the degree of drinking water pollution in the towns and countryside of the Bydgoszcz District, supplied by various installations, on the basis of the results of investigations of sanitary-epidemiological stations. Evaluation was done according to the state in 1985 as compared with that of 1981 to establish the eventual dynamics of the changes. The data presented in the tables indicate that the water supplied by the big water lines (public and institutional) does not contain nitrate quantities exceeding the norm (above 10 mg N/dm3). Their presence at a 20 mg N level/dm3 was found only in four (1.5%) lines supplying large state farms. Small installations were much more frequently polluted with nitrates, especially public wells and those of local institutions, a total of 8.8% in towns and as many as 17% in the country, where a tendency to deterioration of this state is also visible in contrast to the towns. Among the plants and institutions supplied by these water sources the situation is most unfavourable in agriculture, schools, educational and food producing and distributing establishments. The nitrate concentration in the analysed water varied in general within the limits of 10-30 mg N/dm3, maximal amounts within 60-200 mg N/dm3, showing an about 50% decrease in the compared time periods. Nitrates occur most frequently and in highest concentrations in water of wells belonging to individual households which supply about 50% of the rural population and about 11% of town dwellers.(ABSTRACT TRUNCATED AT 250 WORDS)


[Three cases of methemoglobinemia following nitrite intoxication]. Three children of nitrate oral intoxication with symptoms of meta-hemoglobinemia and respiratory insufficiency have been described. All cases have been recovered.


[Detection of carcinogenic compounds in the environment with special regard to tests on fruit flies (Drosophila melanogaster)]. The increasing exposure of the human population to mutagen and carcinogenic substances necessitates research and introduction of methods for their detection. The diversity of these dangerous substances in the environment requires, the application of the chemical and biological tests. The latter allow testing of chemical mixtures of unknown composition. The advantages and limitations of short-term tests are presented. Among these the advantages of somatic mutation and recombination tests are described in more detail on Drosophila melanogaster. The subcellular structure, enzymes produced and the course of metabolic processes in Drosophila are similar as in man. It is possible, therefore, to detect the numerous mutagens undetectable in tests on bacteria. These test are relatively simple, not expensive and do not require expensive apparatus or preliminary activation of promutagens necessary in tests on bacteria.


[Effect of histamine application on diamine oxidase activity in kidneys and histamine level in urine]. Histamine or cadaverine or both amines jointly were given to guinea pigs for 7 weeks. The histamine content was determined in urine and the diamine-oxidase activity in the kidneys. Histamine excretion by the kidneys occurred 4 hours after introduction of the amine into the stomach. In the animals receiving histamine the activity of diamine-oxidase was greatly enhanced.


[Determination of formaldehyde using the colorimetric method with acetylacetone. I. Determination of formaldehyde in selected cosmetic products]. Total formaldehyde content was determined by the colorimetric method with acetylacetone in shampoos and foam baths. The commercial products containing formaldehyde were fortified with a solution of this compound of various amounts. Recovery in this method was 91.7-98.7%. The method was found useful in routine determinations of formaldehyde in shampoos and foam baths and can be used by the State Sanitary Inspection.


[Occurrence of bacteria of the Yersinia genus in surface water]. The aim of the study was determination of the frequency of occurrence of Yersinia genus bacteria in surface waters polluted to various degrees with bacteria of the coliform and of fecal coli. For detection of Yersinia rods the previously elaborated medium Endo MLCe and the membrane filter method were applied. Samples of 42 surface waters were examined, including 26 from rivers and 16 from lakes, ponds and clay-pits. On the basis of sanitary bacteriological analysis 16 surface waters were classified to class I purity, 10 to class II, the remaining ones to class III or beyond classification. Yersinia rods were detected in 15 water bodies that is 35.7% of the examined waters. A total of 27 Yersinia strains were identified with dominance of Y. intermedia (14 strains) and Y. enterocolitica (10 strains). Three strains represented by the species Yersinia frederiksenii. Most of the Y. enterocolitica strains belonged to biotype 1, the particular strains being represented by various serotypes. Hence their different origin may be concluded. The pathogenic serotypes 0:3 and 0:9 of Yersinia enterocolitica were not detected.


[Evaluation of school milieu as an aspect of conditions for realizing health promotion. I]. Data collected in 1990 by workers of children and youth hygiene section of the Sanitary Epidemiological Station were analysed in randomly chosen elementary schools of 41 districts. In elaboration of the data negative observations were singled out. There were differences between schools of various milieus. A regularity was noted: the smaller the locality, the more there are shortcomings and neglect. Teaching in shifts was general: in at least two relays in 90% of town schools and 50% of village ones. Many shortcomings were noted in the functioning of WC and washing equipment (lighting, heating, ventilation), besides 10% of village schools had no washing facilities. About 60% of town schools had showers bath but they were hardly utilised; 1/4 of village schools had shower baths but did not profit of them. In the sanitary-hygienic state frequent negligence was visible. For instance in 1/4 of what used to be communal schools the WC were dirty, without toilet paper, and less than half of them had neither soap nor towels. Lousiness indexes were as follows: 1.6% (if 140,000 individuals examined) in town schools and 2.5% (of 65,000 examined). Lousiness was recorded in more than 60% of schools. The staff of the schools health service may be described by the following data: in 85% of village schools the physician appeared at most several times in the course of the year, in town schools he was present at most several times in a month; in 50% of village schools a nurse was seen less frequently than one a week, and in 85% of town schools three or more times weekly.(ABSTRACT TRUNCATED AT 250 WORDS)


Erythro-toxicity of dichlorvos and possibility of its modification in vitro. The purpose of this study was to investigate the in vitro effects of dichlorvos (organophosphorous pesticide) on the biological value of erythrocytes; an in vitro test of erythrocyte protection against DDVP effects was also included. A significant decrease of glutathione level with concomitant increase of methemoglobin and denatured hemoglobin levels, as well as a rise of Heinz body number were observed after 24 h following the treatment with dichlorvos (100 microM). Preincubation of erythrocytes with glutathione prevented the toxic effects of the pesticide with the maintenance of both glutathione and methemoglobin at a control level.


[The influence of DDT analogues (nuarimol and fenarimol) on the frequency of occurrence of micronuclei in erythrocytes of bone marrow and spleen in mice]. The clastogenic potential of the two DDT analogues: fenarimol and nuarimol was investigated, and the usefulness of the mouse spleen cells for this purpose was evaluated. Nuarimol and fenarimol were administered per o to 8-10 weeks old male mice (Swiss) according to the following protocol: nuarimol 100 and 200 mg/kg b.w. and fenarimol 50 and 100 mg/kg b.w. twice during 24 hours. Mitomycin C (2 and 4 mg/kg b.w.) and cyclophosphamide (25 and 75 mg/kg b.w.) were administered to the positive control mice. After 30, 48, 72 hours following the first administration the animals were killed and the number of micronuclei were calculated in spleen and bone marrow polychromatic and normochromatic erythrocytes. The results show that the spleen could be regarded as an appropriate tissue for the evaluation of micronuclei induction. Fenarimol and nuarimol did not cause micronuclei induction in the bone marrow and spleen erythrocytes.


[Studies of certain trace elements in selected internal organs and meat of bulls]. The aim of the study was to determined lead, cadmium, copper, zinc, and iron contents in certain internal organs and meat of young bulls. The material comprised kidney, liver and muscle tissues of 60 bulls from an agricultural region (control group). Contents of lead, cadmium, iron, copper and zinc were determined by atomic absorption spectrometry. It was found that the levels of lead and cadmium in kidney and liver of the bulls from Kraków voivodship, as compared with the agricultural region, were several times higher.


The Canadian four-centre study of anaesthetic outcomes: I. Description of methods and populations. The objectives of this study were first to develop and institute a methodology for the study of anaesthetic outcome for parallel use in four teaching hospitals in Canada and second, to compare rates of morbidity and mortality associated with anaesthesia between the four centres. The basic design of the study was occurrence screening with anaesthetists entering data on patient demographics, anaesthetic and surgical factors. Research nurses reviewed anaesthetic records and hospital charts and interviewed patients postoperatively. Data on 37,665 anaesthetics were collected during 1988-89 in the four teaching centres. There were major differences found across the hospitals, particularly with regard to volume, patient case-mix, anaesthetic drugs and monitoring used. The use of parallel training, repeated consultations and use of rounds and inservices contributed to the reliability and validity of the data collection. We conclude that outcome surveillance can be instituted in different hospital Departments of Anaesthesia with sufficient confidence to form the basis of comparison of anaesthetic outcome.


Age as a factor in breast cancer knowledge, attitudes and screening behaviour. To determine whether there are age-related differences in knowledge, attitudes and behaviour with respect to breast cancer and whether the differences reflect the age-specific Canadian recommendations on breast cancer screening. Telephone survey. Two cities and five towns and their surrounding areas in Alberta. The age-specific, randomly selected sample comprised 1284 women aged 40 to 75 years who did not have breast cancer. Of the 1741 eligible women who were contacted, 1350 (78%) agreed to participate; 66 were excluded because of age ineligibility or a history of breast cancer. Frequency of knowledge, attitudes and behaviour with respect to breast cancer, by age group. Knowledge of breast cancer risk factors was generally low and decreased with age. Few women were aware of the Canadian recommendations on breast self-examination, physical examination of the breasts by a health care practitioner and mammographic screening. Older women believed they were less susceptible to breast cancer than younger women and were less likely to have positive attitudes toward screening. Self-examination was performed 9 to 15 times per year by 424 women (33%), and 810 (63%) had been examined by a health care professional in the past year. Although 664 (52%) had undergone mammography, the proportion decreased with age after age 59. The main barriers to mammography were lack of physician referral and the woman's belief that the procedure is unnecessary if she is healthy. Education is needed to increase breast cancer knowledge, promote the Canadian recommendations for early detection of breast cancer and decrease negative beliefs about the disease. Changes in the behaviour of women and physicians are needed to increase the use of breast self-examination, clinical breast examination by a health care professional and mammographic screening. Reaching women in the upper range (60 to 69 years) of the target group for mammographic screening should be a focus in promoting early detection of breast cancer.


Direct thrombosis of aneurysms with cellulose acetate polymer. Part I: Results of thrombosis in experimental aneurysms. The authors have developed a liquid material for thrombosing aneurysms. This material is a mixture of cellulose acetate polymer and bismuth trioxide dissolved in dimethyl sulfoxide. On contact with blood, the dimethyl sulfoxide diffuses and cellulose acetate polymer forms, which balloons when slowly injected into the blood. The polymer solidifies from surface to core in 5 minutes. Cellulose acetate polymer was injected directly into experimental aneurysms created in 10 dogs; it rapidly hardened in the shape of the aneurysms, completely obliterating them but preserving the parent vessels in all cases. No distal migration of the polymer was seen. The good results of this experimental trial led to a clinical study using a cellulose acetate polymer, as described in Part II.


Inappropriate medication prescribing in skilled-nursing facilities. To quantify the appropriateness of medication prescriptions in nursing home residents. Prospective, cohort study. Twelve nursing homes in the greater Los Angeles area. A total of 1106 nursing home residents. The appropriateness of medication prescriptions was evaluated using explicit criteria developed through consensus by 13 experts from the United States and Canada. These experts identified 19 drugs that should generally be avoided and 11 doses, frequencies, or durations of use of specific drugs that generally should not be exceeded. Based on the consensus criteria, 40% of residents received at least one inappropriate medication order, and 10% received two or more inappropriate medication orders concurrently; 7% of all prescriptions were inappropriate. Physicians prescribed a greater number of inappropriate medications for female residents. Regression analysis, corrected for clustering effects within facilities, showed that a greater number of inappropriate medication prescriptions were ordered in larger nursing homes. Inappropriate prescriptions were not related to the proportion of Medicaid (Medi-Cal) residents or the number of physicians practicing in the homes. Inappropriate medication prescribing in nursing homes is common. Female residents and residents of large nursing homes are at the greatest risk for receiving an inappropriate prescription.


The influence of standard of care and severity of injury on the resolution of medical malpractice claims. To explore how frequently physicians lose medical malpractice cases despite providing standard care and to assess whether severity of patient injury influences the frequency of plaintiff payment. Retrospective cohort study. Physicians from the state of New Jersey insured by one insurance company from 1977 to 1992. A total of 12,829 physicians involved in 8231 closed malpractice cases. Physician care and claim severity were prospectively determined by the insurance company using a standard process. Physician care was considered defensible in 62% of the cases and indefensible in 25% of the cases, in almost half of which the physician admitted error. In the remaining 13% of cases, it was unclear whether physician care was defensible. The plaintiff received a payment in 43% of all cases. Payment was made 21% of the time if physician care was considered defensible, 91% if considered indefensible, and 59% if considered unclear. The severity of the injury was classified as low, medium, or high in 28%, 47%, and 25% of the cases, respectively. Severity of injury had a small but significant association (P < 0.001) with the frequency of plaintiff payment (low severity, 39%; medium severity, 43%; and high severity, 47%). The severity of injury was not associated with the payment rate in cases resolved by a jury (low severity, 23%; medium severity, 25%; and high severity, 23%). In malpractice cases, physicians provide care that is usually defensible. The defensibility of the case and not the severity of patient injury predominantly influences whether any payment is made. Even in cases that require a jury verdict, the severity of patient injury has little effect on whether any payment is made. Our findings suggest that unjustified payments are probably uncommon.


Effective dose from radiopharmaceuticals. The effective dose, as defined by the International Commission on Radiological Protection (ICRP 1991), provides a possibility of expressing the radiation risk to patients undergoing different radiodiagnostic procedures by means of a single figure. This has been obtained by introducing organ or tissue weighting factors reflecting the radiation sensitivity of the organs. Such weighting factors were first published by the ICRP in publication 26 (1977), and have now been revised in publication 60 (1991). The effective dose for almost all radiopharmaceuticals in clinical use has been recalculated using the new weighting factors from ICRP 60 (1991) and compared with results from former calculations. A slight decrease in the numerical value for the effective dose has been observed, on average 11%. However, this does not correspond to a decrease in the estimated risk from the irradiation, since this has been re-evaluated and found to be higher than earlier believed (NAS 1990; ICRP 1991).


Afferent and efferent connections of the paramedian reticular nucleus in the brain stem of cats. Anatomical connections of the paramedian reticular nucleus (PRN) of the caudal medulla were investigated using a bi-directional tracer, horseradish peroxidase (HRP). The followings were observed when the tracer was microinjected to PRN: A. Both labelled neurons and terminals were found in the areas of the mesencephalic cardioinhibitory mechanism (CIM), the gigantocellular reticular nucleus (GRN), the ambiguus nucleus (AN) and the contralateral PRN. B. Only labelled terminals were demonstrated in the area of the nucleus of solitary tract (NTS) and the intermedial lateral cell column (IML) of the spinal cord. C. Only retrogradely labelled neurons were observed in the areas of the dorsal and dorsomedial medulla (DM) and ventrolateral medulla (VLM). A few labelled neurons were observed in the periaqueductal gray, the cuneiform nucleus and superior colliculus of the mesencephalon as well as the alamina spinal trigeminal nucleus. When HRP was applied to the CIM, GRN or AN structures, respectively, both labelled cells and terminals were found in the PRN area. HRP injection in the VLM showed only labelled terminals in the PRN. However, injection of HRP to DM showed neither labelling neurons nor terminals in PRN. Results suggest that PRN projects to the pressor area of DM/NTS and IML through which PRN could exert its inhibitory functions on the sympathetic pressor actions. In addition, PRN may suppress the vagal bradycardiac action through its reciprocal connections with CIM, GRN and AN. No lateralization in the PRN pathway was evident.


Epidemiology of dengue/dengue hemorrhagic fever in Malaysia--a retrospective epidemiological study 1973-1987. Part I: Dengue hemorrhagic fever (DHF). Dengue hemorrhagic fever (DHF), though endemic in the sixties, emerged as a major public health problem in Malaysia from 1973 onwards. The incidence rate of DHF which was 10.1 per 100,000 in 1973 has fallen down to 1.9 per 100,000 in 1987 with a mean case fatality rate of 6.4 per 100 persons. The Chinese appear to be more prone to DHF with the highest mean morbidity rate of 5.5 per 100,000 and case fatality rate of 6.1%. The incidence of DHF is higher in the males with a higher case fatality rate in females. Male Chinese appear to be mainly affected. The overall age-specific incidence rate is highest in two age groups, viz. 5-9 years and 10-19 years of age with a mean morbidity rate of 4.9 cases per 100,000. The mean age-specific case fatality rate was highest in the 0-4 years age group. Dengue hemorrhagic fever is predominantly an urban disease in Malaysia with a mean incidence rate of 5.3 cases per 100,000 as opposed to 1.2 cases per 100,000 being reported from rural areas. The mean overall incidence of deaths in the urban area is 0.5 compared to 0.1 per 100,000 for rural areas. There is a marked seasonal correlation between DHF cases and rainfall, with a peak in August. While all four serotypes of dengue viruses are found in Malaysia, Den 2 appears to be isolated with greater frequency during all the epidemics.


The managerial process as a key to successful breastfeeding for promoting infant health in Thailand. A strengthening of the managerial process within the health care delivery system can facilitate and encourage successful breastfeeding. This process includes problem-based planning, multidisciplinary team approach, leadership development and generation training. The process has proved to be very successful in promoting breastfeeding and improved infant health. It can be applicable to other urban settings throughout Thailand and perhaps to other developing countries.


The interaction of lead exposure and pregnancy. The toxic effects of low-level lead exposure have been the subject of a good deal of research and media attention in recent times. In most countries, the acceptable occupational exposure limit for lead is being progressively decreased as the adverse health effects of lead are being identified at levels approaching those found in non-occupational environments. Due to the sensitive nature of the fetus to hazardous substances, the exposure to lead of the unborn child via maternal sources is of critical concern. Preterm delivery, congenital abnormalities and decreases in growth stature have all been associated with prenatal lead exposure at "acceptable" levels. There is an accumulation of evidence which indicates that maternal exposures prior to conception can play an important role in determining blood lead levels during pregnancy. In light of these observations the practice of removing the pregnant woman from lead sources may be of questionable value with regards to providing sufficient protection for the fetus. This article reviews the relevant literature pertaining to the mobilization of lead from bone during pregnancy and the toxicity of low-level lead exposure to the fetus, and briefly discusses some factors which may affect this toxicity.


Environmental biotechnology: biotechnology solutions for a global environmental problem, hazardous chemical wastes. Biotechnology has a growing place in the remediation of hazardous waste sites throughout the world, and especially in Asia where population density is high and land and fresh water are scarce. In-situ bioremediation has been demonstrated already to be highly effective for petroleum hydrocarbons (alkanes, aromatics, polychlorophenols) and organophosphate pesticides in soils and for gasoline by-products (benzene, toluene, xylene) and chlorinated solvents (trichloroethylene) in groundwater. Heavy metals and PCBs are not suitable for bioremediation. Environmental biotechnology includes solid-phase and slurry-phase bioremediation for contaminated soils and site-specific bioreactors for contaminated groundwater. Specific examples are presented. From a policy point of view, accumulated wastes must be detoxified, preferably at sites where they already exist. We cannot continue to rely on their removal and disposal "elsewhere". For current waste streams, we must minimize the volumes and toxicity. Environmental biotechnology will play a key role.


Clinical studies on juvenile, rapidly progressive and adult periodontitis before and after periodontal therapy (I). The sample in the present study comprised 36 patients who were referred for treatment of periodontal disease. Twenty-four of them were early onset periodontitis (EOP) patients and 12 were adult periodontitis (AP) patients. Based upon the age, number of involved teeth and the location of diseased sites, the EOP patients were divided into two groups as juvenile periodontitis (JP) and rapidly progressive periodontitis (RPP). All the patients were individually examined both clinically and radiographically. At a baseline examination and 3 months after active therapy, the patients were evaluated for plaque, gingivitis, probing depths and probing attachment levels. Following instruction in proper oral hygiene measures, all of the patients were subjected to periodontal treatment. Treatment comprised scaling and root-planing, occlusal adjustment, tetracycline administration and periodontal surgery with osteoplasty. Results showed that age limits between JP and RPP cannot be drawn with certainty. The mean age for the JP group was 21.83 years and was 30.58 years for the RPP group. The number of involved teeth was found to increase with age. The first molars were the most frequently affected teeth followed by the incisors. Three months after active treatment, the response of periodontal tissues to therapy both in EOP and AP groups was almost identical.


Diagnostic studies on juvenile, rapidly progressive and adult periodontitis before and after periodontal therapy (II). The use of antibody levels may identify subjects developing destructive periodontitis or provide markers of disease type. Clinical parameters can be used to group cases but they are incapable of detecting differences in host-susceptibility and host immune response. The validity of radial immunodiffusion (RID) as a laboratory screening method in diagnosis based on antibody quantitation has been evaluated in 36 patients before and after periodontal therapy. It was concluded that the serum IgA, IgG, IgM and parotid IgA concentration profiles, measured with RID have little value in determining the subgroups of early onset periodontitis patients but they can be used to differentiate these groups from slowly progressing adult periodontitis patients.


Attempted suicide in Kuala Lumpur.

Attempted suicide in Kuala Lumpur. A total of 306 patients were admitted to the University Hospital in Kuala Lumpur in 1989 after attempting suicide. Fourteen of them succumbed to injuries. Psychosocial data of 296 patients out of the 306 survivors are reported. Suicidal behaviour is more common in the young and especially amongst the females. Nearly 45.0% of them are from social class IV and V. Persons of Indian ethnic origin are overrepresented, while in Malays suicidal behavior seemed to be less common. Self-poisoning was reported to be the commonest method in attempting suicide. Diagnosis of adjustment disorder was made in 58.5% of the patients. Two-thirds of the patients had an intention score of less than 10 on the Pierce's Scale.


The fit of all-ceramic crowns cemented with different luting agents. In this article the fit of all-ceramic crowns, cemented with different luting agents (zinc phosphate and resin cements), was investigated using a scanning electron microscope. The marginal opening of all-ceramic crowns cemented with zinc phosphate cement was 111.9 +/- 40.6 microns which was significantly greater than the 46.7 +/- 12.9 microns marginal opening of Cerestore crowns cemented with resin cement. Total film thicknesses of the cements were not uniform and there were certain discrepancies from margins to walls and to tip due to the die spacer.


Pacific health: an analysis for training new leaders. History, economics, culture, and politics have created health systems that can no longer meet the health challenges in the Pacific. An analysis of the health situations in the region revealed health trends, health service deficiencies and the various issues that need to be addressed. The morbidity and mortality experience in the Pacific has been approaching that of the developed countries. The contributing factors to diseases and death have been changing but other health issues, such as matters pertaining to environmental sanitation, management, health information, food and nutrition, ethnic pluralism, colonialism, and technological arrogance, may exacerbate these experiences. To appropriately meet these constraints, the health services need reorientation. However, these need new types of health leadership. Doctors, being the most socially acceptable and politically powerful, should be trained for this new leadership role. This training must be guided by the biopsychosocial model to produce community health physicians. This is being undertaken at the Pacific Basin Medical officer Training Program, Pohnpei. It is characterized by an integrated, community-oriented, student-centered, career-ladder and competency-based approach. The graduates will then provide the leadership and catalyst for the needed health service reorientation.


The incidence of mandibular premolars with more than one root canal in a Turkish population. One hundred and eighty seven mandibular premolars taken from a Turkish population were examined in this study. The teeth were soaked in India ink and then decalcified, cleared and used in an in vitro study to determine the number of root canals and their types, the ramifications of root canals, the location of apical foraminae and the frequency of apical deltas. It was found that the incidence of more than one canal in mandibular premolars was 27.76%; that of finding two or three separate canals in the same root was 9.62% and that of having more than one canal at the apex was 23.49%.


The effect of a sealant on the microleakage of composite resin restorations: an in vivo study. The aim of this study was to evaluate the effect of sealant application in reducing microleakage of the composite resin restorations under in vivo conditions. Occlusal composite resin restorations were made in 24 caries-free primary molars which were about to exfoliate, in children aged between 10 and 13 years. Of these restorations, 12 were sealed with a fissure sealant whereas the others were kept unsealed as controls. After 1 month, the teeth were extracted and immersed in a 2% methylene blue dye solution. The dye penetration alongside the restorations was evaluated using a stereo-microscope. The findings indicated that sealant application significantly (p < 0.05) reduced the microleakage of composite restorations but did not eliminate it completely.


Effectiveness of a worksite nutrition education activity in a factory in China. To evaluate the effectiveness of a worksite nutrition education activity in China, a two-group pretest-posttest nutrition education lesson was designed and carried out in two divisions (N = 240 in each division) of a steel tube factory in Chengdu, Sichuan. Special features of the program were nutrition education materials developed from both (a) the results of a pretest survey of the employees, and (b) focus group discussions conducted with factory hospital and workteam staff; a colloquial style; a slogan; and an illustrated handout depicting the new Chinese Dietary Guidelines and a proposed Food Guide. Analysis of variance, chi-square, and t-tests showed both significant increases (p < 0.05) in nutrition knowledge and attitude scores and significant improvements in dietary practices in the group receiving the education. It is concluded that the method is a useful and practical model for designing and developing worksite nutrition education in China.


Fiber optic surface fluorometry-reflectometry technique in the renal physiology of rats. Most current knowledge on events in the mitochondria leading to acute renal failure originates from studies in which indirect methods were used. The disadvantage of these methods is that they cannot measure the turnover rate of various metabolites, and only one result per animal can be obtained. Chance et al. /9/ developed a method using optical techniques for continuously monitoring the fluorescence of intramitochondrial NADH, which has been applied mainly to the brain. This optical method has not yet been examined quantitatively in the kidney and no attempt has been made to adapt this method for routine measurement in kidney tissue. The purpose of this study was to adapt the surface fluorometry method for monitoring renal NADH redox state in situ, and to determine whether the hemodynamic artifacts involved in fluorometric studies of the renal surface in situ could be eliminated by using a correction factor. Another purpose was to understand the relationship between the changes in reflectance and blood volume in the rat kidney. This was achieved by measuring the reflectance after: a) blood exchange by FC-43 emulsion; b) intrarenal saline flush; c) occlusion of the renal vein, renal artery and reopening of the renal vein; d) calculation of the correlation between changes in kidney weight after renal artery occlusion, and the reflectance. Our results suggest that in the rat kidney, as opposed to the brain, a correction factor of 1:1 is not always applicable. This factor may vary between animals, and it is therefore necessary to adjust it electronically for each rat kidney. This observation contradicts the view suggesting a constant correction factor of 1:1 in the kidney. The results reported herein indicate that changes in the reflectance in the ischemic rat kidney are due to changes in blood volume. In conclusion, it seems that optical techniques for monitoring fluorescence are suitable for localized, continuous and non-invasive recording of tissue mitochondrial NADH redox states under various conditions in the rat kidney.


Streptococcus mutans in plaque from margins of conventional and high copper amalgam restorations. Levels of Streptococcus mutans in plaque samples from margins of conventional and high copper amalgam restorations were compared in fourteen patients, each having one conventional amalgam filling in each quadrant and one high copper amalgam filling on the contra-lateral side were examined. The percentage of S. mutans of total CFU count in plaque was higher on conventional amalgam than on high copper amalgam. The difference was statistically significant (p < 0.01).


The prevalence of Candida albicans in complete denture and removable partial denture wearers: a comparative study. In this study, 60 complete denture wearers, 53 removable partial denture wearers and 50 dentate subjects were examined to determine the Candidal carrier state. The influence of local factors such as denture type, smoking habits and sex on Candidal carrier rate were investigated. Wearing complete or removable partial denture was determined as an important factor increasing Candidal carrier rate. Cigarette smoking is another factor which increases Candidal growth in complete denture and removable partial denture wearers. In contrast to denture wearers Candidal carrier rate was considerably less on dentate smokers. Candidal colonization rate was found to be higher on the dorsum of the tongue. This suggests that the tongue is the primary oral reservoir of Candida albicans in the mouth.


A study of chromium, nickel and cobalt hypersensitivity. Hypersensitivity reactions to chromium, nickel and cobalt were investigated in the preclinical, fourth year and fifth year students of the Dentistry Faculty of Marmara University, Türkiye. A Standard patch test was performed. Four patches were applied to the shoulder blades of the students. Three of them contained small amounts of 0.5% potassium chromate, 5% nickel sulphate and 1% cobalt chloride respectively. The fourth (control) patch was of petroleum jelly. Questionnaires were used to determine sex and history of any previous allergy to jewelry. After 48 hours, the patches were removed and results assessed. For each base metal, no statistically significant difference was found in the incidence of hypersensitivity reaction between the groups. Nor was there a significant difference in response found between metals, for all of the students tested. Although women were found to have a higher rate of nickel hypersensitivity than men, no such sex difference was found chromium and cobalt. No significant relationship was found between a history of allergy to jewelry and chromium hypersensitivity, however there was a significant difference for nickel and cobalt hypersensitivity. In confirmation of our tests, significant relationships were found between previous allergy histories and the respective patch test results.


Cephalometric evaluation of the therapeutic effects of the Herbst appliance in the treatment of Class II. Div I. malocclusion. The purpose of this investigation was to assess the therapeutic effects of the Herbst appliance cephalometrically. Twenty patients were chosen from our clinical intake. Care was taken to select patients who had Class II Div I malocclusion and horizontal growth pattern. Skeletal age was the main selection criterion. Patients in the peak growth period were chosen. A Herbst appliance was used on the first ten patients. The other ten patients served as controls. Standardized profile roentgenograms were taken initially and at the end of a six month observation period. The following results were obtained: 1. Normal occlusal conditions occurred. 2. The mandible was positioned anteriorly and lengthened due to the Herbst therapy. 3. The most significant effect on the maxilla was the decrease in the 1- SN angle and in overjet reduction.


Antibacterial effects of ofloxacin, clindamycin and sultamicillin on surgical removal of impacted third molars. This study examined the bacteraemia following surgical removal of impacted mandibular third molars and evaluated the antibacterial effects of Ofloxacin, Clindamycin, Sultamicillin, used as prophylactic medications. The study involved a hundred healthy patients whose mandibular third molars were impacted horizontally. These patients were divided into four groups each including 25 individuals. One of the four groups was the control group. The other groups were those to which Ofloxacin, Clindamycin, and Sultamicillin were administered one hour before the operation and in the following 4 days postoperatively. Blood samples were taken before and immediately after the operation, and then, 1 and 24 hours postoperatively. Following the incubation of the samples under aerobic and anaerobic conditions, the samples were examined microbiologically. Preoperative samples were found to be negative. In the immediate postoperative samples, bacteraemia was found in 44% of the control group, 40% of Ofloxacin and Clindamycin groups and 36% of the Sultamicillin group. In the samples taken 1 hour after the operation, bacteraemia was found in 28% of the control group, 20% of the Ofloxacin group and 24% of the Clindamycin and Sultamicillin groups. In the control group, only 2 cases showed positive culture in the blood samples taken 24 hours after the operation. In conclusion, the antibiotics, Ofloxacin, Clindamycin, Sultamicillin have a significant effect in decreasing the risk of postoperative infection and bacteraemia.


Effectiveness of three cavity varnishes in reducing marginal microleakage of amalgam restorations: an in vitro study. Dentin Adhesit, Cavi-Line and Kavite Lak cavity varnishes were applied to the walls of prepared Class V cavities in fifty-two caries free extracted molar teeth before placing amalgam fillings. Tests with Methylene Blue dye after thermocycling showed that all cavity varnishes showed significantly lesser marginal leakage when compared with the control group without any cavity varnish.


Evaluation of the variation in chemical composition and surface topography of different brands of gutta percha cones. There have been many studies on the gutta percha cones used in endodontic therapy. Variations exist between different brands of cone. Since an appropriate cone plays a very important role in the prognosis of therapy, this study examined the surface topography and chemical composition of eight different brands of cone by, a scanning electron microscopy (SEM) and atomic absorption spectrometry. There were some differences between all brands of gutta percha cones.


Biguanide related lactic acidosis: incidence and risk factors. The objective of this study was to evaluate in an open population the incidence and risk factors of biguanide related lactic acidosis. All patients currently treated in the Department of Diabetes and Lipid Metabolism of the Instituto Nacional de la Nutrición and their records were reviewed for the present use or history of administration of biguanides. The study was complemented with a revision of all admissions of diabetic patients to the emergency room during 1987-1990. In the outpatient study, 235 cases were included. No case of lactic acidosis was found. A high percentage of the biguanide treated patients had one or more lactic acidosis related risk factors. In the emergency study, 609 admissions of 273 patients were included. In 17 patients a metabolic non-ketotic acidosis was diagnosed. The frequency of non-ketotic acidosis for the different treatments was: 29.4 cases x 1000 emergency admissions for sulphonylurea treated group, 32 for sulphonylurea plus phenformin treated and 47.94 for type II insulin treated patients. All cases had severe precipitant diseases that can cause lactic acidosis with or without associated biguanide administration. No metformin related cases were found. The conclusions of this study are that biguanides in general and metformin in particular are not associated with a high risk of lactic acidosis. Severe systemic dysfunction associated with intercurrent diseases, frequently observed in diabetic patients, is the main determinant for the appearance of lactic acidosis.


Effects of G-6-PD deficiency, experimentally induced or genetically transmitted, on the sorbitol pathway activity. In vitro and in vivo studies. Aldose reductase catalyzes the NADPH-linked reduction of hexoses to their respective sugar-alcohols, which are involved in the pathogenesis of "sugar-cataracts". In the lenses, the reaction catalyzed by G-6-PD is the source of NADPH supply blocking sugar-alcohol formation and consequently prevents or delays the onset of "sugar-cataracts". We have investigated the effect of G-6-PD deficiency, either experimentally induced or genetically transmitted, on the sorbitol accumulation in whole cells incubated in high glucose media and on the "sugar-cataracts" formation in a galactosemic rat model. We also screened 31 Negro male adults with diabetes mellitus for red cell G-6-PD deficiency. G-6-PD deficiency produced a significant inhibition on sorbitol accumulation in rat lenses and human red cells incubated in 50 mM glucose. In the galactosemic rat model G-6-PD deficiency experimentally induced with acetaminophen delayed the development of cataracts. Finally, two diabetic individuals were G-6-PD deficient and did not show cataracts whereas cataracts were identified in six other diabetic patients.


Oral tolerance impairment in mice with staphylococci-induced wasting syndrome. The effects of staphylococci-induced experimental wasting disease on the immune response of 1 month old CD1 mice were investigated. Animals were separated into different subgroups in order to study their cellular immune competence. Malnutrition of wasted mice was associated with anorexia, diarrhea and weakness. Footpad-injected splenic cells produced normal graft vs. host (GvH) reactions in the popliteal lymph nodes of F1 hybrids. In other experiments, the SRBC intragastric feeding of wasted mice did not cause a tolerant anti-erythrocyte humoral immune response. Three weeks after the staphylococci injections, convalescent erythrocyte-fed wasted mice exhibited an anti-SRBC PFC production similar to that observed in non-tolerant immunized control healthy mice. In return, healthy SRBC-fed control mice showed the specific attenuation of antibody response characteristics of oral tolerance. Differences were found between the immune competence of the undernourished staphylococci-treated wasted mice and those shown by other authors in protein-depleted mice.


Sperm motility improvement: comparison of two methodologies of reproductive techniques. The objective of this study was to compare two methods for the extraction of high motility sperm; washed semen and swim-up techniques, on 172 ejaculates from 53 patients with suspected infertility. High quality motile sperm had been essential for the success of the assisted reproductive procedures, in which these patients were enrolled. Semen analyses were performed according to the criteria recommended by the WHO. In both methods Ham's F-10 medium plus 7.5% human cord serum were used. The incubations were carried out at 37 degrees C +/- 1. The percentage of motile spermatozoa with forward progression was measured in the fresh sample and after incubation with both methods. The results were analyzed by the two-way analysis of variance. Motility rates were significantly higher in swim-up than fresh samples (mean 41.158%, SD 20.1 vs. 27.572%, SD 16.6; p < 0.0001) and washed semen compared with fresh sample (mean 36.966%, SD 16.3 vs. 27.572% SD 16.6; p < 0.0001).


Densitometric validation of nine anthropometric prediction equations of body composition in young native men of Mexico City. Body density by hydrostatic method and anthropometric variables were measured in 29 young males, natives of Mexico City, in order to test the validity of nine currently used equations for predicting body composition. Tested equations were those reported for body density by Sloan, Wilmore, Jackson and Pollock, Durnin and Womersley, Lohman, and Pollock et al.; for lean body weight by Wilmore, and Hume-Rathbun; and for fat percentage by Yuhasz. Mean values for body density, fat percentage and lean body weight obtained by hydrostatic weighing were 1.0560 +/- 0.0149 kg/l, 18.9 +/- 6.7%, and 52.6 +/- 5.3 kg, respectively, while the means for predicted body density using the six regression equations varied from 1.0538 to 1.0654 kg/l, with a range of correlation from 0.846 to 0.900, and total errors from 0.0077 to 0.0124 kg/l. Mean lean body weight predicted by Wilmore was 55.7 +/- 5.9 kg, with an R of 0.902 and a total error of 4.0 kg, while values given in the same order by Hume-Rathbun were 52.8 +/- 4.8 kg, R of 0.892 and total error of 2.3 kg. Mean fat percentage predicted by Yuhasz was 17.8 +/- 5.6%, R of 0.869 and total error of 3.5%. Although by analysis of variance there were no significant differences among the means of the models, statistical analysis of individual differences between predicted and measured values were significant in six of them.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of a PGE-1 analogue on the survival of cardiac transplant rats. Prostaglandins can be considered as soluble factors of cell-mediated immunity. Studies with animal models have shown that prostaglandins of the series E-1 (PGE-1) can modulate the lymphocyte response to alloantigens. The goal of this work was to evaluate the immunosuppressive effect of a PGE-1 analogue (Enisoprost) on cardiac allograft survival in rats. PGE-1 was given to groups of six rats with heterotopic cardiac transplant. Group 1 was the untreated control group. Group 2a received PGE-1 from 0 to 4 days after transplant and group 2b received PGE-1 from 7 days before transplant to 4 days after transplant. Group 3 was treated with donor-specific blood transfusions (DST) 7 days before transplant. Groups 4a, 4b, and 4c were treated with DST and PGE-1 (0 to 4 days, -7 to 0 days, and -7 to 4 days relative to transplant, respectively). Group 5 was treated with cyclosporine (CsA), groups 6a, 6b, and 6c received DST, CsA and PGE-1. Cardiac allograft survival of group 2a (PGE-1) was better than that of the control group (9.6 +/- 1.7 days vs. 6.6 +/- 1.9 days) X +/- SD; p < 0.05. Group 4a (DST+PGE-1) had better cardiac allograft survival than group 3 (DST) (20.0 +/- 14.3 days vs. 14.3 +/- 3.3 days, respectively; p < 0.05). Group 6a (DST+CsA+PGE-1) had a better graft survival than group 5 (CsA) but the difference was not significant (44.6 +/- 13 days vs. 37.5 +/- 19.5 days, respectively; p = 0.20).(ABSTRACT TRUNCATED AT 250 WORDS)


Hypoglycemic effect of plants used in Mexico as antidiabetics. The objective of this work is to investigate the hypoglycemic effect of 12 "antidiabetic" plants used in Mexico. The studies were performed using 27 healthy rabbits with the gastric administration of water, tolbutamide or decoction of the "antidiabetic" plant before the induction of temporary hyperglycemia by subcutaneous injection of 50% dextrose solution (4 ml/kg of weight) at the beginning of the experiment and after 60 min. Blood glucose was determined every 60 min for a period of 5 h. Tolbutamide and eight of the studied plants decreased significantly the hyperglycemia as compared with control test (water) (p < 0.05). The strongest effect was yielded by Guaiacum coulteri, followed by Marrubium vulgare, Crataegus pubescens, Cynodon dactylon, Calea zacatechichi, Buddleia americana, Bauhinia divaricata and Coix lachryma. The decrease of hyperglycemia caused by Physalis phyladelphyca, Pavonia schiedeana and Eucaliptus globulus was not significant (p > 0.05). Urtica dioica increased glycemia slightly.


Hodgkin's disease and acute leukemia. Is it a true association? The incidence of secondary malignancy was assessed in 537 patients with Hodgkin's disease treated with radiotherapy (128 patients), chemotherapy alone (156 patients), or in combined modality therapy (253 patients) between January 1973 to September 1985 with a median follow-up of 7.5 years. The dose of radiation therapy, dose of cytotoxic drugs and sequence of treatment was carefully analyzed. No cases of acute leukemia or other secondary malignant disease were identified in these cases. No differences in clinical laboratory features or treatments were identified in relation to previous reports. Racial difference is the unique feature which seems to avoid the development of this complication in patients treated for Hodgkin's disease. We hope that other reports in Latin America can contribute to the identification of race as the difference.


Local and systemic antibody response in Balb/c mice immunized with Entamoeba histolytica trophozoites. Several immunization schedules with E. histolytica trophozoites were tested on Balb/c mice in order to induce antibody responses, both in intestinal secretions and in serum. Mice were immunized either orally, systemically, or using one of two combined schedules: the oral route followed by the systemic route (footpad), or vice versa. Each of the immunization schedules used in this project induced an anti-E. histolytica antibody response and there appears to be a correlation between the immunization route employed and the immunoglobulin isotype induced in the gut. Secretory IgA production is favored by the oral administration of trophozoites, whereas mucosal IgG appears to be enhanced by the systemic immunization route. Both schedules are effective in the induction of secretory IgA in the gut, yet higher and earlier levels of IgA appear in orally immunized mice. When systemic immunization is employed, the increase in antibody levels in the intestinal fluid is slower, and IgG is the predominant class. The combined oral/systemic routes of immunization appear to be comparably effective for the induction of local and systemic IgA and IgM antibody production. However, mice immunized first systemically and then locally produce more IgG in both compartments. Combined schedules modify the isotype pattern of antibody responses in serum and in intestinal secretions when compared with single (i.e., oral or systemic) schedules, but they do not appear to favor a secretory IgA immune response.


A study of erythropoiesis and iron metabolism in the rabbit in vivo. II. Dependence of the response on iron storage and transport. Rabbits subjected to a daily bloodletting schedule of 10 ml blood per kg body weight increase four- or fivefold their erythropoietic production compared to normal non-bled animals. The maximum response they can reach under these conditions mobilizes more than 9 mg of iron per day per rabbit into hemoglobin. When fed ad libitum with their regular diet, they do not need any further iron supplement for full erythropoiesis. The experimental increment in iron body stores and/or serum iron levels does not enhance their erythropoietic response, demonstrating that iron is not rate limiting under the conditions studied. Furthermore, although serum iron levels are elevated onefold in the controls under chronic anemia with respect to non-bled animals, the concentration of serum transferrin is only slightly increased; hence, the iron saturation of this protein changes from a 50% to an 80% level. In the absence of an extra supplement of iron, rabbits subjected to chronic bloodletting show no signs of body iron depletion, as judged by their continuous macrocytic RBC production.


Verapamil does not block the spastic response of Praziquantel on the larvae of Taenia pisiformis. This paper describes that the previous addition of Verapamil does not block the spastic response to Praziquantel (Pz) in larvae of Taenia pisiformis kept in vitro, where opposite results were found in the literature using mammalian tissue. It is possible that Pz stimulates other Ca++ transport channels not sensitive to Verapamil action and promotes Ca++ liberation from calcareous corpuscles stimulating phospholipase C of the tegument surface generating inositol triphosphate. These hypotheses require experimental approaches to define the exact mechanism of action.


Hypoglycemic activity of some antidiabetic plants. To assess the hypoglycemic activity mechanism of some plants used empirically by the Mexican population as antidiabetics, traditional preparations of Cucurbita ficifolia, Guaiacum coulteri, Lepechinia caulescens, and Psacalium peltatum, water, tolbutamide, and Regular Insulin were administered to three groups of rabbits each: 1. Healthy rabbits with temporary hyperglycemia induced by the subcutaneous administration of glucose. 2. Rabbits with moderate diabetes (fasting glycemia 150-300 mg/dl), induced with alloxan. 3. Rabbits with severe diabetes (fasting glycemia higher than 400 mg/dl), induced with alloxan. The plant preparations had a hypoglycemic effect similar to tolbutamide in healthy and mild diabetic rabbits and had no effect in severely diabetic rabbits. These results suggest that some pancreatic function or the presence of insulin is required for the hypoglycemic activity of these plants.


Plants used by Mexican traditional medicine with presumable sedative properties: an ethnobotanical approach. An ethnobotanical study of plants used in Mexican traditional medicine was made. The source was the national inquiry done by the IMSS-COPLAMAR health program (1983-1985) in which the plants used to treat mental disorders were selected and analyzed, in order to select the most frequent botanical species used in traditional medicine as sedatives, anticonvulsants and hypnotics.


Efficacy of pravastatin as a hypolipidemic agent in patients with polygenic hypercholesterolemia. In Mexico, hypercholesterolemia has become a major public health problem particularly in the states of the north of the country and in Mexico City, where a prevalence of 20% has been reported. Schemes of treatment have now been reinforced by the appearance of new cholesterol reducing drugs. The objective of the study was to demonstrate efficacy and safety of a 10 mg daily dose of oral Pravastatin (a new 3-hydroxy-3-methyl glutaryl CoA inhibitor) in a group of patients positive for hyperlipidemia, after 6 months of treatment. Twenty-five patients were included (14 men, 11 women) with an average age of 54 and 50 years, respectively. The main outcome measure was total cholesterol (T-CHOL), low density lipoprotein-cholesterol (LDL-C), triglycerides (TGL), high density lipoprotein cholesterol (HDL-C) and adverse drug reactions report. Twenty-one out of 25 patients completed the study. T-CHOL diminished 21%, LDL-C was reduced by 28%, TGL decreased 6% and HDL-C increased 32%. No adverse reactions were observed throughout the study. Our study shows that the use of a low dose of Pravastatin satisfactorily reduced T-CHOL and LDL-C levels while significantly increasing HDL-C after 27 weeks of treatment, without untoward effects.


EEG signs of "relaxation behavior" during breast-feeding in a nursing woman. The presence of electroencephalographic signs of "relaxation behavior"; i.e., groups of synchronic, 6-10 Hz, 100-150 microV EEG waves mainly appearing in parieto-occipital regions, have been investigated in a nursing woman during breast-feeding. EEG was recorded once a week from right and left frontal-central, central-parietal, parietal-temporal and parietal-occipital derivations of the mother during breast-feeding of her own child throughout 16 weeks of nursing (from the 4th to the 20th week after delivery). Groups of synchronic 6-10 Hz, 100-150 microV EEG waves were recorded in central-parietal and parietal-temporal derivations, 15-80 sec after starting of suckling of either breast. Proportions of this synchronic EEG activity ranged between 5 to 28% in the central-parietal and 6 to 22% in the parietal-temporal derivations, and significantly higher proportions were found in the right hemisphere. Similarities between EEG signs of "relaxation behavior" elicited by suckling in nursing cats and those elicited by suckling in a nursing woman lead to the possibility that the above described EEG phenomena are an expression of a functional state of the central nervous system which is a part of the integral response involved in the consummatory activity of nursing behavior in women.


Osteochondral lesions in developing rats intoxicated with thallium twenty four hours after birth. An i.p. injection of a solution of thallium acetate in deionized water at a dose of 32 mg/kg, in 24-h-old rats, produces morphological and biochemical alterations in both cartilaginous and osseous tissues. From the beginning, there are alterations in the cartilaginous cell as well as in chrondrine, osteoblasts, osseous tissue and bone marrow. Rats were sacrificed at 24, 48, and 72 h and also at 7 days. Two animals survived for 50 days. One showed total irreversible alopecia while the other one had partial alopecia with discrete recovery. Both showed a low weight and a size of 8 cm. Microscopically, degenerative changes were produced consisting of alteration and death of many cartilaginous cells, uneven metachromasia and the chondrine and decrease of the growth cartilage, scanty bone trabeculae with few osteoblasts. The bone marrow showed few myeloblasts and megakaryocytes. Progressive cellular damage throughout the 50 days of survival represents a response of the thallium ionic accumulation and recycling in cellular mitochondria of all the body's cells. This appeared in our study as irreversible and progressive osteochondral alterations with atrophy of the skin and its adnexa, hyalinization of elastic and collagenous fibers with intense interstitial edema.


Perinatal prevalence of B hepatitis markers. The present investigation describes the prevalence of markers detected in immunoenzymatic methods, for infection by the hepatitis B virus in sera of mothers and newborn umbilical cord blood in Merida, Venezuela. The sample population was composed of 500 mothers who attended the Obstetrics Department, University Hospital of los Andes, for their delivery between May and October 1990. The global prevalence figure of 11.8% showed a balanced distribution for surface antigen and its antibody. Prevalence for the urban area was 13.49% (RR = 1.49) and 11.80% for mothers from the rural zones (RR = 0.67), 95%, IC 0.63 to 1.9). A statistical difference was found between mothers with two or more live children (14 cases) and those with one or no children (7 cases) for anti-HBs (X2 = 7.62, p < 0.05, RR = 1.43, CI 0.47 to 2.86). Mothers with a history of several sexual partners showed a prevalence of 18.0%, RR = 1.5, CI 0.58 to 2.44, while those who were students, 18.3%, RR = 1.64, CI 0.94 to 2.5. These data may be indicating that sexual transmission mechanisms are of utmost importance, yet more precise investigations are required to confirm this hypothesis. Blood samples from umbilical cord were all negative for surface antigen.


Comparative study between the effects of total intravenous anesthesia with propofol and balanced anesthesia with halothane on the alveolar-arterial oxygen tension difference and on the pulmonary shunt. Inhalation anesthesia with halothane, inhibiting hypoxic pulmonary vasoconstriction, causes an increase in intrapulmonary shunt development as well as an increase of alveolar-arterial oxygen tension difference. In contrast, total intravenous anesthesia with propofol, as with other intravenous anesthetics, should not alter pulmonary gas exchange. The present study was carried out using two groups of patients of similar age, sex and weight, who were subjected to neurosurgical procedures. One group was under general inhalational anesthesia with halothane and the other group under total intravenous anesthesia with propofol. In a simple clinical manner and applying established formulae, the determination of intrapulmonary shunt and alveolar-arterial oxygen tension difference showed a significant increase after 120 min in the halothane group, whereas the group that received propofol did not show any significant variation. We therefore conclude that propofol, different from halothane, does not affect the pulmonary gas exchange.


Thyroid function and abdominal surgery. A longitudinal study. Surgical trauma produces complex metabolic changes and may be an excellent model for studying the effects of an acute insult on the circulating thyroid hormone concentrations. Abnormalities in thyroid function tests in patients with nonthyroidal illness can be divided into a low T3 syndrome, low T3 and T4 syndrome in more severe cases and elevated T4 syndrome. In this study 30 patients undergoing elective or urgent abdominal surgery were longitudinally evaluated with pre- and postoperative thyroid function tests. Comparing with preoperative values, a significant reduction in total T3 serum values in the immediate postoperative period was seen (p < 0.005). The rest of the variables did not show significant statistical differences between the three periods. Over half of the patients undergoing urgent surgery had a low T3 syndrome, whereas only a sixth of the patients scheduled for elective surgery suffered from this syndrome. In the late postoperative period half the patients submitted to urgent surgery persisted with alterations in thyroid function tests, whereas most of the patients scheduled for elective surgery showed an improvement in their thyroid hormone level in the same period of time. Abnormalities in thyroid hormone metabolism were more frequent in patients admitted for urgent surgery, reflecting the severity of their illness and favoring the concept that these syndromes can be used as prognostic markers.


Immune response to parasitic infection in mice without seminal vesicles. A protracted depression of anti-parasitic antibody and DTH responses were observed in Balb/c mice after surgical extirpation of seminal vesicles and ulterior infection with Taenia crassiceps cysts. Inclusion of male seminal accessory glands into the network of immunogonadal interactions was proposed.


Capacitation and acrosome reaction of human spermatozoa. An assisted reproduction approach. In this paper we briefly review the recent advances in the understanding of human sperm capacitation, fundamentally focused to give help to the practitioners involved in assisted reproduction. Sperm capacitation has been the subject of study for many years in several animal species, but during the past ten years the advances in assisted reproduction procedures have made important contributions in the progress of this topic in relation to human spermatozoa. In this manuscript we analyze the metabolic, membranal and physiological changes observed in these cells during in vitro capacitation and their relationships with the female genital tract secretions, as well as with the homologous oocytes.


Molecular cloning and sequence analysis of the cDNA for northern pike (Esox lucius) growth hormone. The complete nucleotide sequence of the northern pike (Esox lucius) cDNA for pregrowth hormones was determined from clones derived from a pituitary gland cDNA library. Seventeen cDNA clones were isolated from a single mRNA species. A cDNA of 1,227 nucleotides was sequenced and found to encode a polypeptide of 209 amino acid residues, which included a putative signal sequence of 22 amino acid residues. Sequence comparison of the northern pike growth hormone gene to other known growth hormone genes revealed similarities closest to other members of the superorder Protacanthopterygii, which includes the Salmonidae family (i.e., salmon and trout).


Characterization of AluI repeats of zebrafish (Brachydanio rerio). Two families of repetitive DNA sequences were isolated from the zebrafish genome and characterized. Eight different sequences were sequenced and classified by two standards, their (G + C) composition and their lengths. For convenience, the sequences were first divided into two types. Type I was (A + T)-rich, was repeated approximately 500,000 times, and constituted approximately 5% of the zebrafish genome. Type II was (G + C)-rich, was reiterated approximately 90,000 times, and comprised approximately 0.5% of the genome. Agarose gel electrophoresis of zebrafish DNA cleaved with AluI revealed three distinguishable bands of repetitive fragments: large (approximately 180 bp, designated RFAL), medium (approximately 140 bp, RFAM), and small (approximately 90 bp, RFAS). The RFAL fragments contained both type I and type II sequences. Limited digestion of genomic DNA indicated that RFAL and RFAM were tandemly arranged in the genome, whereas RFAS showed a mixed pattern of both tandem and interspersed repeated arrangements. Although inclusion of a repetitive sequence in a transgenic construct did not appreciably accelerate homologous integration of transgenes into the zebrafish genome, the AluI sequences could facilitate transgene mapping following chromosomal integration.


Automated sequential affinity chromatography of sea urchin embryo DNA binding proteins. An automated method of running a tandem sequence of oligonucleotide affinity columns was used to purify factors that interact specifically with cis-regulatory sites of the CyIIIa cytoskeletal actin gene of the sea urchin embryo (Strongylocentrotus purpuratus). The method allows quantitative enrichment in a single chromatographic run of up to 12 different sequence-specific DNA binding proteins, each of which may then be readily purified to homogeneity by methods such as preparative gel electrophoresis. The affinity chromatography and identification of six different CyIIIa-regulatory factors is described, and the general utility of the method is discussed.


A gene encoding chinook salmon (Oncorhynchus tschawytscha) prolactin: gene structure and potential cis-acting regulatory elements. A full-length chinook salmon (Oncorhynchus tschawytscha) prolactin (PRL) gene, the first genomic clone of a teleost prolactin, was isolated and fully sequenced. The chinook PRL genomic sequence spans 6.4 Kb, including 2.4 Kb of 5' flanking sequence, 3.0 Kb representing the five exons and four introns of the complete PRL gene, and 0.9 Kb of 3' flanking sequence. The transcriptional start site of the PRL gene was mapped through the agreement of both primer extension and S1 nuclease protection assay. The 5' flanking region of the PRL gene was searched for potential cis-acting elements based on the consensus binding site of trans-acting factor Pit-1, known to be involved in PRL gene expression in mammals. Functional analysis of PRL promoter by the transient transfection of several PRL promoter/CAT chimeric plasmids into rainbow trout pituitary cells suggests a functional PRL promoter whose cell-specific activity is most likely governed by both positive and negative mechanisms.


Isolation and sequence analysis of carp gonadotropin beta-subunit gene. Using the cDNA encoding the beta subunit of carp gonadotropin (cGTH-beta) as a probe, 14 clones containing cGTH-beta gene have been isolated from a carp genomic library. Nucleotide sequence analysis indicated that the transcriptional unit of the cGTH-beta gene is 1.2 Kb. Similar to mammalian GTH-beta genes, cGTH-beta gene contains three exons and two introns. The locations of the exon/intron junctions also correspond to those of mammalian GTH-beta gene. Using the primer extension assay, the start site of transcription was determined to be 35 or 37 bp upstream from the translation initiation codon. The TATAA box is present in the 5' flanking region of the gene, 21 bp upstream from the start site of transcription. Three polyadenylation signals, AATAAA, are located in the 3' noncoding region, 111, 430, and 442 bp downstream from the stop codon of translation, respectively.


Integration of chromosome set manipulation and transgenic technologies for fishes. Chromosome set manipulation techniques have significant implications for research of transgenic fish. Gynogenesis can be used to generate isogenic lines, to map genes in relation to their centromeres, and to produce fish carrying extra chromosome fragments of foreign origin. Androgenesis can be used to produce isogenic lines and to recover strains from cryopreserved sperm. Triploidy can be induced in fish with heat or pressure treatment of fertilized eggs and by crossing tetraploid individuals with normal diploids. Triploid fish are typically effectively sterile. Triploid interspecific hybrids are usually more viable than the corresponding diploid hybrids. Given concerns about potential reproduction of transgenic fish in the wild, induced triploidy could facilitate application of transgenic technologies in some situations.


Fish embryo cell cultures for derivation of stem cells and transgenic chimeras. It has been demonstrated in mammalian systems that techniques using embryonal stem cells provide advantages over conventional injection of DNA into embryos for generation of transgenic animals. We employed cell culture approaches in an attempt to develop this technology for fish transgenesis. Using a trout embryo-derived mitogenic preparation in a specialized culture medium, we initiated replication of zebrafish blastula-derived cell cultures and expressed marker genes introduced into the cells by plasmid transfection. Reintroduction of cells from the cultures into blastula-stage embryos indicated that the cultured cells survived and may contribute to the developing organism.


Expression of reporter genes introduced by microinjection and electroporation in fish embryos and fry. The technique for foreign gene transfer in fish is becoming a novel method for genetic engineers to produce useful transgenic fish as well as for experimental purposes. Our studies of transgenic fish have been focused on methods of gene transfer and regulatory elements for transgene expression. I describe the characteristics of 3 gene transfer methods we have established (i.e., microinjection into fertilized eggs, microinjection into oocytes, and electroporation). Also described is a series of experiments to estimate activities of several promoters and enhancers in a fish cell line. Finally, experiments to examine activities of the elements shown to be active in the cell line in fish embryos and fry are described. SV2, miw, and metallothioneine promoters of trout and mouse were shown to be active in these experiments.


Introducing foreign genes into fish eggs with electroporated sperm as a carrier. A new method has been developed for introduction of foreign genes into fish eggs. The procedure is based on the incubation of fish sperm cells suspended in dilute citrate solution with plasmid DNA, followed by application of high-field-strength electrical pulses (electroporation) to increase DNA binding., uptake, or both. Tissue homogenates and genomic DNA extracts of free swimming fry developed from eggs fertilized with treated sperm was tested to evaluate the efficiency of gene transfer. Dot blot hybridization and gene expression assay demonstrated the presence and expression of the reporter genes introduced in 2.6 to 4.2% of several hundreds of tested larvae of common carp (Cyprinus carpio L.), African catfish (Clarias gariepinus), and tilapia (Oreochromis niloticus). No transgene has been found in the fry resulting from parallel experiments without sperm electroporation. This is the first report on successful application of electroporated sperm cells for production of transgenic fish.


No transgenic rainbow trout produced with sperm incubated with linear DNA. We attempted to produce transgenic rainbow trout embryos by fertilizing eggs with sperm incubated with linearized plasmids. One experiment was conducted with the construct pBGH7 in the medium MMSF, with or without DMSO, at 2 concentrations of sperm cells and a relatively low concentration of DNA. The DNA was also in contact with the eggs during insemination and during the first minutes of egg activation. The second experiment was conducted with the construct CMVCAT in the medium MMSF, at 2 concentrations of sperm cells and a much higher concentration of DNA. The DNA was also present during the insemination. DNA analyses and dosages of CAT activity did not permit detection of any transgenic fry. However, one result suggests that sperm cells can capture part of the linear DNA in teh conditions tested.