[Study on the meiotic mapping and functions of another new osmotic-sensitive gene-osm3 in Saccharomyces cerevisiae]. Another recessive nuclear gene mutation of Sacchromyces cerevisiae, osm3, which causes growth inhibition on hypertonic media, has been mapped on the chromosome II by tetrad analysis. Its second-division segregation frequency is about 51.01%, and the map distances between osm3 and the centromere of chromosome II is about 25.51 centimorgans and it is approximately 45 centimorgans far gene gall. The studies on the reverse mutation of osm3 indicated that this osmotic-sensitivity arises from a missense or nonsense mutation in OSM3 locus. The response of yeast to osmotic stress and the mechanism of osm3 gene function were investigated. The results indicated that high intracellular glycerol content is necessary for yeast to grow in hypertonic media; The gene product of osm3 is probably relative to the process of glycerol transportation. Finally, the effect of osmotic stress on fermentability was also discussed.


[Construction and characterization of shuttle plasmid pBHG1 in Bacillus thuringiensis and Escherichia coli]. The recombinant plasmid pBHGA was constructed by ligating plasmid pHTA1030 from B. thuringiensis to plasmid pJH101 from E. coli. The recombinant plasmid pBHGA was digested and deleted by various restriction enzymes, generating 14 derived recombinant plasmids differing in molecular weight. The expressible analysis for the plasmid pBHG1 from among the derived plasmids in E. coli HB101 and B. subtilis 168, the results proved the recombinant plasmid pBHG1 is carring promotive region, original region and segregational stability region of B. thuringiensis plasmid pHTA1030. The recombinant plasmid pBHG1 can be high frequency transformation in to B. thuringiensis HD-1 Cry- B and high-expression in B. thuringiensis. HD-1 Cry- B. The plasmid pBHG1 is segregational stability in B. subtilis 168 at 45 degrees C. These plasmids may be useful tools for study of plasmid replication and expression in B. thuringiensis once a reproducible transformation system is available for this organism.


[The correlation between the maturation of rbcL transcripts and its flanking sequence in a prokaryotic system]. There is a 15bp large reverse repeated sequence proceeded by a 7bp small one in the 3'flanking region of rbcL of Nicotiana tabacum. A 383 bp of XbaI fragment containing these tandemly repeats was inserted into the plasmid p lambda S delta, at the position between the lambda p and the cat gene. Then these two repeats were separated and deleted systematically to obtain various deletions. The deletion pRT65, pRT74 and pRT83 was sequenced to determine the deleted base pairs exactly. S1 mapping analysis was adopted to investigate the transcripts of these deletions in E. coli JM83. The results showed us that the stability of mature 306bp mRNA relied on the large repeat and a short sequence downstream. The small one was not efficient. The regulation level of the rbcL termination was also investigated. The XbaI-EcoRI fragments from pRT65, pRT74 and pRT83 were transferred into pSP-TT* at the position between the spinach promoter and threonine terminator to construct pRT65, pRT74 and pRT83 respectively. The results from S1 mapping analysis showed that the E. coli RNA polymerase read through the 3' flanking region of rboL gene and terminated at the stop site of threonine terminator. These results suggested that mature rbcL mRNA might be the product precisely processed from a precursor mRNA and the 3' flanking sequence might be the signal for precursor mRNA to be processed to the correct position.


Effects of short length immobilization of medial gastrocnemius muscle of growing young adult rats. Effects of four and six weeks of immobilization at short length of gastrocnemius muscle on its architecture at optimum muscle length and length-force characteristics were studied. In general, immobilization effects were similar after 4 and 6 weeks. Smaller physiological cross-sectional area and lower muscle force were found as a consequence of immobilization. Muscle and aponeurosis were shorter. This was shown to be quantitatively related to atrophy i.e. smaller fibre diameter. Despite this atrophy no effects of immobilization on fibre and aponeurosis angles could be shown. Adaptation of the number of sarcomeres in series was found exclusively in distal fibres after 4 weeks of immobilization. No significant effects were found for proximal fibres of muscles at this time nor for any fibres after 6 weeks of immobilization. The effects of immobilization on muscle architecture did not affect the length range of active force exertion. It is concluded that muscle length adaptation as a consequence of short length immobilization is not related to adaptation of number of sarcomeres in series but to the occurrence of atrophy. It is also concluded that atrophy of pennate muscles does not have to be accompanied by a lower fibre and aponeurosis angle. Comparison of immobilized and control group rats indicates that the effects of immobilization can be characterized as a combination of retarded development of several variables and the influence of atrophy and its consequences.


Pericytes in human breast stroma and the cells to which they relate. The function of pericytes, the cells nearest the microvascular endothelium, has long been debated. On the basis of ultrastructural studies it is pointed out that they have specialised features in common with endothelial cells of the lymphatic labyrinth of the human breast. The latter are in continuity with the initial lymphatics. These features, which include points of stromal attachment, allow contraction or relaxation of the cell-processes. The cytoplasmic processes of the pericyte, when relaxed, form areas of contact with the blood vascular endothelium. Subsequent contraction may lead to loss of contact and increase in the area of vascular endothelium available for diffusion. The pericyte is thus equipped to act as a regulatory link between the blood vascular endothelium and that of the fine lymphatic system.


Tissue and sex differences in the expression of nucleoli in mouse somatic cells. In Mus musculus, the nucleolus organizer regions (NORs), or sites of ribosomal RNA-encoding genes, map at three chromosomal pairs. A silver procedure was modified to stain nucleoli in interphasic somatic cells of mice. The number of nucleoli per cell nucleus was determined in squashed cells of kidney, liver and pancreas obtained from male and female mice. In liver and pancreas cells the average number of nucleoli per cell was 4.84 and 4.66, respectively, and only 2.83 in kidney cells (p < 0.001). Less than 8% of pancreas cells and about 15% of liver cells contained more than 6 nucleoli per cell, which was the maximum expected number. In addition, the number of nucleoli per cell was significatively different (p < 0.01) when male and female liver or pancreas cells (not kidney cells) were compared. In both cases, female cells presented more nucleoli than the respective male cells. Assuming that the available NORs are the same, the variable number of nucleoli in the examined cell types would be the consequence of a tissue specific NOR regulation. The apparent influence of sex on this regulation is noted.


Calibrating microwave-incubation times for droplet immunostaining: reversible Thermo-Paint as a low cost alternative. Reversible Thermo-Paint R50 (Manufactured by Thermographic Measurements, Ltd., Burton, UK and imported in The Netherlands by Blanken Controls, Loenen (Vel.)) has a yellow colour that changes to orange when a temperature of 50 degrees C is reached. Microscopical glass slides can be painted with R50. Under the droplet on the painted slide an orange disk will appear at the moment the droplet has reached the temperature of 50 degrees C. This test provides us with information about how much energy per minute is available to be absorbed by that precisely defined "test droplet" using a certain setup of the microwave oven. The obtained data can be used to calibrate incubation times for various microwave ovens. The results explain why, when short cycle times are used, immunostaining is highly reproducible.


[The content of the nursing consultation at a health center]. To describe the care process in the Nursing Station. To analyse the length of a consultation and factors which affect the length. Crossover study. Analysis of the length of the Consultation, using a multiple lineal regression model. SITE. Urban Health Centre. PATIENTS AND OTHERS TAKING PART: All the calls and consultations, both in the patient's home and in the Health Centre, undertaken by six nurses over a period of two weeks. There were 879 consultations. 65.41% of them took place in the treatment room. Ten health problems represented 72% of the work-load. Health education took un the greatest proportion of time. The average time of a consultation was of 10.40 +/- 0.3 minutes. The length of the consultation depended on the professional involved, on the place where it took place, on the number of problems which the patient had and on the nature of the main problem: multiple correlation coefficient = 0.74; F = 67.52 (p < 0.00000001). The nurse faces a very limited number of health problems. Most of her activities are preventive or to do with prevention. The length of the Consultation in greatly conditioned by the amount of morbidity.


[An analysis of the obligatory reporting of meningitis in a pediatric area]. To find out the validity, reliability and quality of the epidemiological information which comes from the System of Statutory Declarations. The example used is the incidence of Meningitis. Descriptive study of the coverage of the Declaration, with the help of the EDO (Illnesses requiring a Statutory Declaration) index, of the pediatric (up to 14 years old) cases of meningitis attended in our Area between 1984 and 1989. We analysed how the Declaration had been filled in and the agreement between the information gathered through the notification system and clinical records. SITE. The whole network of epidemiological vigilance: hospitals, provincial epidemiological services and primary care services. All the cases of child meningitis (120 cases) seen in the Mother and Child Hospital in our area between 1984 and 1989. The meningitis declaration covered 47.5% of the total number of cases seen. In the cases declared, the diagnosis of meningococcal meningitis on the EDO index was 89.5%, whereas the rate in the clinical reports was only 20%. Thus agreement in the diagnosis does not exceed what could be produced by chance (Kappa = 2.5%; p = NS), as agreement for all diagnoses is 7.1%. Our results demonstrate a low level of declaration, poor standard of compliance and scant agreement between the Statement of Declaration and the clinical report. The validity and reliability of this information is extremely low and suggests a need to re-consider the usefulness of this vigilance system for health-service practice, especially in Primary Health Care.


[The attitudes of primary care professionals on mental patients and psychiatric care]. To describe the views of different primary care professionals regarding mental illness, psychiatric patients and mental health care. Crossover survey. SITE. Primary care centres in a health zone. All the primary care centre professionals and social service base teams. Out of 66 possible subjects, 61 answered the questionnaire. A questionnaire composed by ourselves about peoples' opinions was given. The chi 2 test was then used to analyse the questionnaire, both item by item and by means of the distribution of frequencies and comparisons. On some items differences between Health Professionals and social service staff occurred. But there were no differences between doctors and nursing staff, nor between men and women (level of importance: 0.50). For 91% of those surveyed, mental health is an illness like any other. 87% believe psychiatry is a science; and 77% that psychiatry is related to the rest of medicine. 48% believe that the majority of mental disturbances are untreatable. 80% affirm that psychiatric admissions should be into special hospitals. The authors highlight the need to overcome the discrepancy between positive attitudes on the theoretical side and a reluctance to integrate those attitudes into practice.


[Tobaccoism in medical students].

[Tobaccoism in medical students]. To evaluate medical students' understanding of the harmfulness of tobacco; their attitude to it; and the spread of tobacco addiction among them. This was a crossover study, using information collected through a self-answer questionnaire. SITE. It was carried out in the Faculty of Medicine in the Autonomous University of Barcelona. The people under study were the 490 students registered in the first year of Medicine and doing the Biostatistics course. The information was collected by means of a voluntary self-answer questionnaire, which was distributed during class-hours of the Biostatistics course. 68% of those registered answered. This broke down into: 25.7% smokers; 25.1% ex-smokers; and 49.2% non-smokers. 88% of students expressed total agreement as to the harmfulness of tobacco, though there were important differences between smokers and non-smokers (p < 0.01). The illness which they related most closely to tobacco addiction was Lung Cancer; and the least closely related was Bladder Cancer. Only 30% would warn healthy smokers about the risks attached to their addiction. 50% believe it is the doctor's responsibility to convince the general population to give up smoking. The spread of tobacco addiction is lower among medical students than among the general population of the same age and than among health professionals. We think it is essential to integrate an action programme against tobacco addiction into the overall study plan.


[An epidemiological study of arterial pressure in a schoolchild population]. To know the prevalence of high blood pressure in childhood. Epidemiological transversal study. SITE. School population, level community. A population of 566 children of 6, 10 and 13 years old of Madrid public schools. The blood pressure measure was registered two times according to the WHO's guidelines. We considered the first sound as systolic blood pressure and the fifth auscultatory phase as diastolic blood pressure. Considering as hypertensive those subjects whose blood pressure are higher than percentile 95 for their age and sex, the total prevalence of systolic hypertension was 5.8% and the total prevalence of diastolic hypertension was 1.4% in both genders. We realized that systolic hypertension prevalence is higher in males for all ages and its is also higher than the results offered by similar studies. It doesn't happen the same thing in the diastolic hypertension prevalence which we found that the measurements registered in males are higher than females but in contrast with other studies we found similar prevalence. We need to identify early the presence of higher blood pressure with the purpose of to prevent in time.


[Lumbago cases seen at a health center]. The main aim is to describe the cases of mechanical lumbalgia in relation to clinical and demographic variables, in the period between February and June 1989. The collateral purpose is to analyse the relationship between analgesic/NAI treatment as regards the time needed to resolve the problem. This is an observational prospective study. The cases included under the second aim are distributed at random. SITE. The study was carried out in the Primary Care Centre of Deltebre, a rural town of 10,000 inhabitants. The consultations for rheumatoid lumbar pain were 355. Of these, 183 were first visits, which were the object of the descriptive study. Out of these, 125 were lumbalgias of the posterior branch and are the target population of the analytical study. Paracetamol (on its own or with codeine) taken orally was used as an analgesic. Piroxicam, Indometazine, Ketoprofen or Diclofenac taken orally, rectally or intramuscularly was used as an anti-inflammatory. MAIN RESULTS AND MEASUREMENTS: In cases between the ages of 40 and 49 in the case of men, and of 40 and 59 in the case of women, an increase in relation to the municipal census was observed, with p < 0.01 and 0.001. There was no difference in the time of resolution between the cases treated with analgesics and those treated with NSAI. Future studies will have to analyse the increase in lumbalgias in the 40 to 59 age-group. The number of cases off-work signifies a health cost. Drug treatment plays no part in the resolution, which agrees with a previous reference.


[The influence of the social factor on the response to a vaccination program]. To make an analysis of the different reasons for the response to three subprograms of anti-tetanus vaccination. Retrospective descriptive analysis. SITE. Within the primary care field: in the rural clinic of Izarra-Urkabustaiz in the province of Alava. Inhabitants who live and/or work in Izarra-Urkabustaiz were allocated for the purposes of vaccination to three groups: 130 workers in a rubber factory, 116 citizens over 65 and 65 people aged 24, 34, 44, 54 and 64. We have used the chi 2 test to analyse the qualitative variables and the confidence interval, in order to find the accurate level of response. We have established at 0.05 the threshold of permitted error and significance. The confidence intervals of the vaccination response have been 74-88 for those over 65, 54-74 in the factory and 4-21 in the "decade" subprogram. The difference has been very significant (p < 0.0001). The factors which best explain such differences are those of a sociological nature, inasmuch as the level of acceptance and response is directly related to the amount of cohesion in the group.


Role of the Office International des Epizooties in protecting the health of free-ranging mammals. The various activities undertaken by the Office International des Epizooties (OIE) since 1924 to protect the helth of free-ranging mammals are outlined and discussed. Two types of activity have been conducted over the years: --The first type promotes measures to protect the health of free-ranging mammals in order to safeguard the health of domestic animals and human beings. These measures have prevented the propagation of the highly contagious diseases of animals and the more serious zoonoses. --The second type aims at protecting the health of free-ranging mammals in order to maintain the fauna and the natural equilibrium. Ecological and epidemiological studies are promoted with a view to effective management of wildlife populations and overcoming the associated health risks. By virtue of its close contacts with officials in charge of animal health in 126 Member Countries, the OIE has made a noteworthy contribution to the protection of populations of free-ranging mammals.


Epidemiology of sylvatic trichinellosis in north-western Italy. Muscle samples from 7,142 wild boars (Sus scrofa), 80 free-ranging hogs, 1,462 red foxes (Vulpes vulpes) 90 mustelids, 15 Alpine marmots (Marmota marmota), 873 rodents and 92 insectivores were examined for Trichinella Railliet, 1895. A wild boar, a badger (Meles meles) and 30 foxes (2.1%) were found to be infected. Eleven vulpine isolates and the wild boar isolate were identified as Trichinella britovi. These results show that the wild boar has little significance as a reservoir of T. britovi in the area under study. A vulpine isolate resulted in low infectivity in experimentally-infected swine. The prevalence of trichinellosis in the vulpine population is higher in the mountains than in lowland areas and indicates the key role played by this carnivore in the epidemiology of T. britovi. Preliminary results from four field trials seem to demonstrate that differences in attitudes towards cannibalism exist between mountain and lowland foxes.


Parasitosis of stray dogs in north-eastern Spain. The authors analysed 42 stray dogs in north-eastern Spain. Of these, 29 dogs were parasitised by one or more helminth species. The parasites found were intestinal nematodes (59.52%) and cestodes (47.61%). Differences between urban stray dogs and feral dogs are discussed.


[Detection of the eggs of Echinococcus multilocularis Leuckart, 1863, in the feces of the fox (Vulpes vulpes Linnaeus, 1758) by the polymerase chain reaction]. The polymerase chain reaction (PCR) was applied to the identification of eggs of Echinococcus multilocularis in faeces from foxes. The test was positive in three of six faeces samples from foxes which were harbouring adult worms, and in one of four samples from foxes in which no adult E. multilocularis was found in the intestines. These initial results show that it is possible to use PCR to identify E. multilocularis eggs in faeces. PCR can be used to complement examination of intestinal contents, showing that the distribution of eggs in faeces is uneven. The sensitivity of the test was estimated to be 50 eggs in 5 g of faeces. Further work is needed to confirm these initial results before the test can be used more widely.


Parasitological and serological studies on the prevalence of Echinococcus multilocularis Leuckart, 1863 in red foxes (Vulpes vulpes Linnaeus, 1758) in Switzerland. In the Canton of Zurich in Switzerland, 1,252 red foxes (Vulpes vulpes) were examined during 1990-1991 for intestinal stages of Echinococcus multilocularis using the mucosal smear technique. Special safety precautions were employed during examination. An average of 35% (432 foxes) were infected, mostly with low to medium numbers of gravid worms producing thick-shelled eggs. In the eleven districts of the Canton, prevalence rates varied between 13% and 57%. An average of 29% of the foxes had antibodies in serum or body fluid against a highly species-specific antigen of E. multilocularis (Em2-antigen). The fact that foxes with intestinal E. multilocularis infection have been found in all parts of the Canton of Zurich indicates a relatively high potential infection risk for humans, but apparently the risk is reduced by certain extrinsic or intrinsic factors which have yet to be determined.


[Histopathologic study of the host-parasite relationship: the earthworm-, wild boar-metastrongyle model]. A histopathological study was conducted on two hosts of metastrongyles (Metastrongylus sp.): earthworms and wild boars (Sus scrofa Linnaeus, 1758). In the earthworm intermediate host there were rare granulomas outside the normal site occupied by larvae (blood sinuses of calciferous glands). Given the small number of these reactions, they could not constitute a limiting factor for parasitism in this host. However, the lungs of wild boars contained numerous lesions associated with the presence of these nematodes. Such lesions, often inflammatory, were part of an immunological defence mechanism which controls the extent of parasitism in wild boar.


Dermatophilosis in camels (Camelus dromedarius Linnaeus, 1758) in Kenya. Natural dermatophilosis (caused by Dermatophilus congolensis Van Saceghem, 1915) has only recently been described in camels (Camelus dromedarius). Further work has shown that the disease is actually widespread. At the Ol Maisor farm where it was first diagnosed (in the Laikipia district of northern Kenya), detailed investigations have revealed that thirty camels (ten calves and twenty adult animals) were infected. The signs varied from mild to more than 50% skin involvement. Patches of wool on the rump were easily detachable with many hard, white crusts. Similarly, another twenty camels from a totally different area of northern Kenya (Samburu district) were found to be infected with D. congolensis. Bacterial isolation was used to confirm the diagnosis. At Ol Maisor farm, twenty sheep were also found to have severe dermatophilosis. There were scabs and many areas of wool shedding. The shed wool had crusts similar to those observed on camels. Experimental infection with D. congolensis isolated from camels was conducted, by means of scarification, on the skin of two goats. The same experimental infection was possible in two calves, taking two weeks to become established. After thirty days there were disseminated foci of infection on the heads of the calves, especially around the eyes. Although these findings cannot show the actual source of D. congolensis, it is possible that the same organism causes dermatophilosis in the different animals.


Myxomatosis: population dynamics of rabbits (Oryctolagus cuniculus Linnaeus, 1758) and ecological effects in the United Kingdom. In 1953-1955, myxomatosis spread among rabbits (Oryctolagus cuniculus) in the United Kingdom, causing 99% mortality. Subsequently, there was a gradual increase in rabbit numbers. By 1955, the Ministry of Agriculture, Fisheries and Food (MAFF) had already found attenuated strains of myxoma virus. By 1970, genetic resistance had appeared. In the 1970s, mortality declined to 47-69% with only approximately 25% of rabbits infected, giving a field mortality of 12-19%. However, myxomatosis is persistent, generally showing a major prevalence peak in autumn and often a minor peak in spring. An eight-year MAFF experiment in which prevalence of the disease was artificially reduced indicates that myxomatosis remains a significant factor in population regulation. After rabbit numbers fell in the 1950s, important ecological changes took place: vegetation altered due to reduced grazing pressure, predators were affected by the reduction of a major prey species and these changes also affected many other animals. Currently, rabbit numbers have returned to approximately one-third of pre-myxomatosis levels and this is causing damage to farm and conservation habitats.


Sarcoptic mange in Swedish wildlife.

Sarcoptic mange in Swedish wildlife. Mange caused by Sarcoptes scabiei var. vulpes appeared among red foxes (Vulpes vulpes) in Scandinavia (south-west Finland) for the first time in this century in 1967. The disease was most probably introduced by foxes crossing the Gulf of Finland from Estonia. The mange epizootic spread northwards through Finland and reached Sweden in late 1975, when mangy foxes appeared in the northern part of the country. In 1984, mange was observed in most parts of Sweden. The disease was observed to spread rapidly in boreal areas, whereas it spread more slowly in agricultural areas. Mortality due to mange was very high. The duration of the disease before death due to emaciation has been shown experimentally to be over 90 days. An outbreak of fox mange among Arctic foxes (Alopex lagopus) occurred in 1986. The local population of Arctic foxes was caught and successfully treated. The following year, treated foxes were caught again and no signs of disease were found. Sporadic cases of fox mange have also been diagnosed in lynx (Lynx lynx), pine marten (Martes martes) and domestic dogs. Single cases have been observed in other species: wolf (Canis lupus), mountain hare (Lepus timidus), domestic cat and horse. No cases of sarcoptic mange have been recorded in the badger (Meles meles). At present, although fox mange occurs as an epizootic in local populations, the number of foxes has increased again in many parts of Sweden.


The ecology of tularaemia.

The ecology of tularaemia. Tularaemia, a zoonotic disease caused by the bacterium Francisella tularensis McCoy, 1912, is reported from North America, Europe and northern parts of Asia, but not from the Southern Hemisphere. Two subspecies of F. tularensis are recognised: the highly virulent type A and the milder type B, with additional subdivisions reported. Tularaemia has been reported in more than 250 animal species including man, other mammals, birds, fish, amphibians, arthropods and protozoa. Type A is reported to have a terrestrial cycle with the main reservoirs being cottontail rabbits (Sylvilagus spp.) and ticks. Type B is reported to have a mainly water-borne cycle with aquatic rodents as reservoirs, e.g. muskrats (Ondatra zibethicus) and beaver (Castor canadensis) in North America, and ground voles (Arvicola terrestris) in the former Soviet Union. In Europe, tularaemia is most frequently seen in hares (Lepus spp.) although hares probably do not constitute a reservoir for the disease. Tularaemia is transmitted by direct contact with infected animals, through contaminated water or food, or by vectors such as mosquitoes or ticks. The disease normally occurs as an epidemic, both in man and in animals, depending on the types of reservoir involved and the means of transmission at different times of the year.


Crassicaudosis: a parasitic disease threatening the health and population recovery of large baleen whales. This communication briefly reviews knowledge of the systemic disease caused by Crassicauda boopis in blue whales (Balaenoptera musculus), fin whales (B. physalus) and humpback whales (Megaptera novaeangliae). Infections with this giant nematode characteristically incite a chronic inflammatory reaction of the blood vessels which drain the kidneys. In this critical location, the parasite-induced lesion can cause complete vascular occlusion and kidney failure. Whale calves and juveniles typically suffer the heaviest parasite burdens following transplacental infection of the developing whale foetus. There is also probable whale-to-whale transmission post-partum, involving urinary contamination of the environment with C. boopis eggs and larvae. The frequency of the infection can exceed 95%. Haematological findings suggest that systemic pathological effects are typical at the population level. Gradual development of occlusive lesions in the renal veins appears to correlate with a major peak in natural mortality at about one year of age. To date, all findings support the conclusion that premature death caused by C. boopis infection is potentially a major impediment to population recovery of affected whale species. This suggests the interesting possibility of actively encouraging the population recovery of three species of large baleen whales. Such a restoration effort would entail remotely-deployed anthelminthic therapy administered, at sea, to infected whale cows and calves.


Epidemiology and control of rabies in bats in The Netherlands. The first case of bat rabies in the Netherlands was recorded in 1987. Since then, 2,540 bats have been examined. In total, 186 bats belonging to two species were found positive for rabies: 182 Eptesicus serotinus and 4 Myotis dasycneme. The virus isolates from the two species were found to be different. Approximately 20% of the E. serotinus bats sent for examination were found positive. The epidemiological findings suggest a stable situation of infection north of the Rhine and the Meuse. A total of 174 humans received post-exposure treatment after direct contact with a positive or suspected bat. The public has been warned not to touch bats and, if contact does occur, to send the bat for rabies examination. The authorities also advise post-exposure treatment for dogs and cats which have had contact with a rabies-positive bat.


Serological survey for bovine bacterial and viral pathogens in captive Arabian oryx (Oryx leucoryx Pallas, 1776). Tests for antibodies to bovine bacterial and viral pathogens were conducted on 239 sera from 128 Arabian oryx (Oryx leucoryx) from seven locations (Taif, Riyadh and Mahazat as Said, Saudi Arabia; San Diego, United States of America [USA]; Shaumari, Jordan; Qatar; and Bahrain). No antibodies to Pasteurella multocida type E or epizootic haemorrhagic disease 1 virus were found. Antibodies to Brucella abortus, P. multocida type B, P. multocida type D, lumpy skin disease virus and Akabane virus were detected in 2, 1, 5, 2 and 1 animals, respectively. Evidence of P. multocida type A, Coxiella burnetti, Chlamydia psittaci and parainfluenza 3 virus was found in 3 herds (prevalence in the main herd [n = 78]: 8%), 3 herds (8%), 6 herds (7%) and 5 herds (15%), respectively. Evidence of antibodies against bluetongue virus was found in five oryx from the USA and in one oryx from the Taif herd. Antibody vaccinal titres against rinderpest virus (and the virus of peste des petits ruminants, due to cross-reactions) were found in almost all the herds. This is the first report of antibodies against B. abortus, C. burnetti, C. psittaci, parainfluenza 3 virus and Akabane virus in the genus Oryx.


Trypanosomiasis in the black rhinoceros (Diceros bicornis Linnaeus, 1758). A black rhinoceros (Diceros bicornis) moved from a tsetse-free to a tsetse-infested area in Kenya was monitored for two months following translocation. The animal acquired a Trypanosoma vivax infection from natural tsetse challenge, but survived without requiring treatment with trypanocides. The infection was characterised by moderately high parasitaemia, with symptoms of anaemia, leukopaenia and thrombocytopaenia. Although confirmed to be T. vivax through deoxyribonucleic acid hybridisation and parasite development in tsetse in the proboscis only, the parasite had unusual morphology and motility. It also failed to infect normally susceptible hosts such as cows and goats, and produced unusually low infection rates in Glossina morsitans centralis and G. brevipalpis.


[Changes in the properties of the 5-HT1 receptors in the brains of rats, the progeny of parents consuming morphine for a long time]. Studies were performed in two-month rats whose parents had been given morphine for a long period before mating. Considerable changes were found in the properties of rat brain 5-HT1 receptors, which manifested as increased number of ligand binding sites (Bmax) and higher KD, which indicates a lower receptor affinity for ligand. Single morphine administration to the animals caused normalization of the properties of the receptors.


[The protective action of nimodipine on the ischemic myocardium]. The experiments with unconscious cats and dogs have demonstrated that the calcium antagonist nimodipine has a profound anti-ischemic property. The drug reduces the average value of ST-segment elevation in multiple epicardial ECG leads, during acute myocardial ischemia. Nimodipine maintains cardiac pump and contractile functions, elevates ATP levels in the arbitrarily intact and ischemic myocardium of the left ventricle during 40-min occlusion and 60-min reperfusion of the coronary artery. The protective action of the drug is unassociated with enhanced collateral coronary circulation.


[The activity of mitochondrial enzymes in the myocardium in chronic coronary failure during the administration of antianginal agents]. The experiments with Chinchilla rabbits indicated that the activity of mitochondria-associated enzymes changed hetero-directionally in chronic coronary insufficiency. The activity of creatine kinase changed most greatly. Obsidan and finoptin were found to show a different effect on the activity of the tested enzymes in the cardiac mitochondria during experimental coronary insufficiency.


[The effect of proanthocyanidins from Alhagi pseudoalhagi (M. B.) Desv on the course of experimental myocardial infarct]. Intravenous proantocyanidin isolated from Alhagi pseudoalhagi (M. B.) Desv, 5-10 mg/kg, has no effects on the general hemodynamics and cardiac contractility of intact rats and rabbits. The agent given to animals with experimental myocardial infarction reduces serum creatinine phosphate levels and lipid peroxidation both in the myocardium and serum, as estimated by the amount of malonic dialdehyde and dienic conjugates), normalizes free fat acid content and cardiac phospholipid spectrum. At the same time the size of a myocardial necrotic zone became diminished.


[The effect of the table salt substitute sanasol on arterial pressure, water-salt metabolism and kidney functions in experimental pathology of circulatory homeostasis]. The potassium-containing salt substitute sanasol was examined for its effects on blood pressure and renal function in normo- and hypertensive rats, as well as on the edematous syndrome in circulatory insufficiency. The antihypertensive and occasional antiedematous effects of sanasol resulted in unfavorable changes in electrolyte balance. Sanasol was found to be ineffective in experimental renal failure.


[The gastroprotective and antiaggregant activities of 13-azaprostanoids]. Some 13-azaprostanoic acid derivatives were shown to have a high cytoprotective activity against the damaging effect of ethanol on the gastric mucosa in non-strain rats and affect ATP- and arachidonate-induced platelet aggregation in the whole blood of rabbits and rats in a different way. The gastroprotective activity that is common to natural and synthetic prostanoids is not closely related to the chemical structure of the compounds. On the contrary, the trends of these compounds to affect the induced platelet aggregation depend on the modification of both the cyclic moiety and both chains of a prostanoid molecule.


[The effect of isradipine on the central and cerebral hemodynamics of patients with circulatory encephalopathy and hypertension]. The new dihydropyridine calcium antagonist isradipine versus nifedipine was tested in 41 patients with essential hypertension and cerebrovascular insufficiency. The tests have shown that isradipine shows all hemodynamic effects by significantly improving cerebral blood flow and it is better tolerated than nifedipine. It is concluded that isradipine is an effective agent in the long-term management of hypertensive patients at a high risk for cerebral hypoperfusion.


[The effect of bonnecor on blood coagulability and thrombocyte aggregation in transient myocardial ischemia]. The experiments on conscious rabbits have indicated that bonnecor given in doses of 0.05 mg/kg and 0.5 mg/kg accelerates clot formation and platelet aggregation, without changing the density of a thrombus. In experimental transient myocardial ischemia, the agent decreases the hypercoagulative change associated with the damaging effect of occlusion and reperfusion.


[The effect of mildronate on erythrocyte membrane permeability in experimental lung pathology]. With the use of the erythrocyte osmotic and acidic resistance methods, the experiments on intact albino rats by applying a model of chronic non-specific lung diseases and those in vitro have shown that mildronate has a membrane-stabilizing action only when given in high doses (500 mg/kg) and concentrations (10(-3)-10(-4) M). When used in low doses (5, 25, 50, and 200 mg/kg) and concentrations (10(-5)-10(-7) M), in produces no positive effect.


[The effect of antidepressants on the reorganization of circadian mobility in normal and stressed rats after a shift in the light regimen]. Changes in the light regime caused a continuous reorganization of circadian motor rhythm with a gradual formation of a new pattern of circadian mobility. Both chronic administration of imipramine and maprotiline and pain stress-induced simulation of depression modified this process in different ways. However, when antidepressants were given, impairments in rhythm resynchronization attenuated in the stressed rats to a great extent.


[The absence of stereoselectivity in propranolol uptake by the lungs]. Propranolol (P) is known to be actively captured by the lungs of experimental animal and man. To elucidate the mechanism of the phenomenon, the authors studied the P enantiomeric ratio in the serum from the left ventricle 0-10, 10-20 and 20-30 s after bolus injection of P (0.5 mg) into the pulmonary artery of 8 patients catheterized for coronary angiography. The total P concentrations and enantiomeric ratio were measured by non-stereoselective and stereoselective high performance liquid chromatography. The solid-phase extraction method was used for extracting P from the serum. It showed a small, but significant increase is S(-)/R(+) ratio (to 1.18 +/- 0.12, P < 0.05 vs 1) which was, however, almost the same when measuring this after adding P to serum of the same patients (1.11 +/- 0.05, P > 0.05 vs 1.18). Thus, propranolol uptake by the lung during a single passage is not stereoselective and hardly involves some active process.


[The metabolism and biological activity of glucuronidation substrates in experimental diabetes mellitus]. The study was undertaken to examine the impact of experimental diabetes mellitus on the biotransformation of some drugs, their toxicity and hypoglycemic action, as well as on the biotransformation of endogenous testosterone and the androgenic status. The accelerated metabolism of both exogenous and endogenous substrates which occurred in diabetes mellitus caused a decrease in their biological activity.


[The possible mechanisms of the interaction of hepatotropic diagnostic agents with proteins]. The phenomenon of precipitation in protein solutions after addition of hepatotropic diagnostic media is described. The examination of different proteins and media allows the coagulative mechanism to be proposed, through which the electrostatic interactions between di- and polyvalent ligands with positively charged protein groups play the major role. It is suggested that the described phenomenon contributes to the nephrotoxicity of some drug groups.


[The effect of desferal on the cellular proliferative activity of the mouse spleen, thymus and bone marrow]. CBA mice were given a single Desferal dose (50 mg/kg). The mice received 3H-Thymidine 1 hour before they were killed. They were killed 1, 2, 6, or 9 hours after Desferal. Radiometry, and autoradiography were used to determine the level of DNA synthesis. Desferal was found to stimulate splenic 3H-incorporation 3 hours after drug injection and to increase the DNA-synthesizing cell fraction. In contrast, the agent inhibited 3H-incorporation in the bone marrow and thymus 9 hours after injection. This suppressive affect was accompanied by a decrease in the DNA-synthesizing cell fraction in these organs. In the thymus, there was a significant increase in the DNA-synthesizing cell fraction among lymphoblasts 1 and 3 hours following the administration of Desferal.


[The nonspecific mechanisms of the pharmacological correction of motoneuron trophic failure in experimental botulin poisoning]. Cobalamines, leukovorine, cymidine phosphocholine, dalargin are pharmacological agents that affect the support of motoneuronal trophic functions, unlike the actoprotector bemityl, prevent the development of fatal botulinic intoxication, but have no effects on the specific mechanisms of botulinic damage when the threshold concentrations of toxin achieve target cells. The depotentiating effect of the drugs is associated with the increased natural degradation of botulinic toxin in its target cells.


[Essentiale as an immunomodulator in a toxic liver lesion]. Tetrachlormethanum used in Wistar rats caused an increase in their immune response to sheep red cells, followed by its decrease. Administration of essential in combination with tetrachlormethanum prevented the accumulation of immunomodulating agents and impairment of an immune response to sheep red cells.


[The therapeutic efficacy of alloxime in experimental carbamate pesticide poisoning]. The new cholinesterase reactivator alloxime, 10 mg/kg, shows a marked therapeutical benefits in acute intoxication rats with carbamine pesticides such as carbofuran, pirimor, elocron. The therapy of alloxime in combination with atropine, 10 mg/kg, results to their combined therapeutical effect, the toxicity of carbofuran, elocron, and pirimor (by LD50) decreasing by 8.4, 5.6, and 3.5 times, respectively). The mechanism of alloxime's therapeutical action is due to its capacity to restore cholinesterase activity in the central nervous system, normalizing neuromuscular transmission and hepatic and renal cytochrome P-450 levels.


[The antihypoxic action of malonic acid]. Malonic acid showed a protective action on rats in acute hypoxic hypoxia. The antihypoxic effect of this compound is unassociated with its action on succinate dehydrogenase and may be mediated through the changes in hormonal metabolic regulation.


[The activities of aldehyde dehydrogenase, GABA-aminotransferase and succinic semialdehyde reductase in the brain of rats with different preferences and tolerances for ethanol]. The formation and catabolism of aldehydes were compared in the hemispheres and brain stem of rats preferring ethanol (EP) or water (WP) and of those which were high tolerant (HT) and low tolerant to the hypnotic effect of ethanol. It was shown that aldehyde dehydrogenase was more active in the brain stem of HT-EP rats than that of HT-WR or LT-EP animals, whereas GABA aminotransferase is most active in the hemispheres and brain stem of LT-EP rats. The total activity of succinic semialdehyde reductase was equal in all the groups studied; however kinetic analysis suggest that the enzyme has a higher affinity for the substrate and coenzyme in the brain stem of HT-EP rats. Ethanol administered to HT-EP animals suppressed aldehyde dehydrogenase in the brain stem, unchanged GABA aminotransferase and activates succinic semialdehyde reductase in the two brain structures.


Combination of interleukin-2 and interferon-alpha in renal cell carcinoma and malignant melanoma: a phase II clinical trial. A total of 22 patients with metastatic renal cell carcinoma or malignant melanoma were treated in a phase II study to assess the safety and efficacy of combination therapy of interleukin-2 (IL-2) and interferon-alpha (IFN-alpha). 3 x 10(6) U/m2/day recombinant human (rh)IL-2 was given in repetitive cycles by continuous 24-h infusion from day 1 to day 4; 6 x 10(6) U/m2/day rhIFN-alpha was given subcutaneously on days 1 and 4. There was one complete remission and two partial remissions in the renal cell carcinoma group and two partial remissions in the malignant melanoma group, giving an overall response rate of 24% in 21 evaluable patients with a median response duration of 5+ months. Toxicity was moderate, with hypotension, fever, chills, nausea, neurotoxicity, and dermatitis as prominent side effects. Measurement of circulating cytokine levels showed increased serum tumor necrosis factor-alpha (TNF), interferon-tau, and soluble interleukin-2 receptor levels during each cycle with a tendency to higher concentrations of TNF in responders as compared to nonresponders. With regard to therapeutic efficacy and tolerance, our approach might represent an alternative to the high-dose protocols and the labor- and cost-intensive strategies of adoptive immunotherapy.


[The effect of phytin, benzonal and their combination administered prophylactically on the pharmacodynamics of drugs metabolized in the liver in hypokinesia]. Hexenal, meprobamate, amidopyrine and ethylmorphine produced a significantly marked effect in animals under hypokinesia as compared with normal rats. When phytin, benzonal and their combination were used for preventive purposes, impaired pharmacodynamics of the tested drugs metabolizing in the liver disappeared. The investigations demonstrated that the preventive use of phytin in combination with benzonal is the most optimal in correcting the impairments of drug pharmacodynamics in hypokinesia.


Muramyl peptide adjuvants for Plasmodium falciparum and Plasmodium vivax circumsporozoite vaccines in rodent model systems. Circumsporozoite proteins from the malaria parasites Plasmodium falciparum and Plasmodium vivax were expressed at high levels in the yeast Saccharomyces cerevisiae. Recombinant proteins varied both in length and in number of the natural amino acid repeat motifs. The proteins were purified and used to immunize mice, guinea pigs, and rabbits. Novel muramyl peptide adjuvants were used that increased the immune response as measured by ELISA assays, indirect immunofluorescence of fixed sporozoites, and the invasion of cultured liver cells by live sporozoites. These results suggest that an improved humoral response to recombinant circumsporozoite vaccines might be achieved by varying the design of the recombinant protein and by the use of novel adjuvant systems.


Trace amounts of murine immunoglobulin in affinity purified leukocyte interferon alpha are not immunogenic. Human leukocyte-derived interferon alfa-n3 (Alferon N Injection) is purified to very high specific activity over a murine immunoaffinity column specific for human interferon alpha. Trace amounts of murine immunoglobulin copurify with the interferon alfa-n3. Three populations of individuals were studied for the development of human anto-murine antibodies (HAMA), that is, normal donors, Condylomata acuminata patients receiving interferon alfa-n3, and Condylomata acuminata patients receiving placebo. High and variable endogenous levels of HAMA were observed in all three populations. The same relative increase in HAMA was seen in the placebo as in the interferon alfa-n3 treatment groups. The data demonstrate that intralesional injection of the interferon alfa-n3 did not induce the development of HAMA.


Conjugated anti-TNF antibodies enhance the binding and degradation of TNF. We have used disulfide-linked conjugates of murine anti-TNF antibodies to determine whether TNF binding to the cell surface could be increased by exploiting the Fc receptors present on monocytes and macrophages. Binding and degradation of TNF via Fc receptors may enhance the ability of anti-TNF antibodies to lower the high TNF levels found in several diseases. Conjugated murine anti-TNF antibodies greatly increased the binding of TNF to human U937 cells and had little effect on cells which did not express Fc receptors. U937 cells treated with anti-TNF conjugates also degraded more TNF than untreated cells. Competition analysis indicated that Fc receptors were involved in anti-TNF binding and that conjugated anti-TNF shared binding sites with monomeric anti-TNF. Preincubation of anti-TNF conjugates or monomeric anti-TNF with TNF increased the amount of anti-TNF antibody bound to the surface of cells. These results demonstrate that cross-linking may greatly enhance the ability of anti-TNF antibodies to mediate TNF clearance and degradation by making effective use of Fc receptors.


IL-2 phase II trial in metastatic melanoma: analysis of clinical and immunological parameters. Twenty-four patients with histologically proven metastatic malignant melanoma were included in a phase II trial of recombinant IL-2 (rIL-2, RU 49637). Twenty million international units (IU)/m2/day were given by continuous intravenous infusion on days 1 to 5, 15 to 18, and 29 to 31, and then monthly for 5 days until disease progression or major intolerance developed. All patients were evaluable for response and toxicity. Toxicity was consistent with one case of myocardial ischemia, 13 cases of grade III and IV hypotension, and 15 cases of proven sepsis. There were 8 objective responses: 4 of them were of short duration as they were observed on day 31 only. An activation of the immune system was detected in all patients. It was demonstrated by an increase in lymphocyte populations, especially in activated NK cells. A tendency for higher numbers of cytotoxic cells was found in patients with objective tumor responses. These results indicate a role for rIL-2 RU 49637 in treating patients with metastatic malignant melanoma. However, further trials are required to determine its optimal dosage and schedule of administration.


[The ultrastructural characteristics of the changes in sections of the rat brain under piracetam exposure]. Rat experiments have shown that intraperitoneal piracetam leads to formation of abundant coated vesicles in the neurons of the sensomotor cortex, hippocampus, and central gray matter. Concurrently, subsuperficial cisterns form under the plasmalemma of nerve cells. The interneuronal contacts are characterized by long-term excitation without destructive changes in synapses. It is suggested that piracetam is able to increase the number of synapses in some brain regions at the expense of sprouting.


Immunotherapy of advanced ovarian carcinomas by activation of the idiotypic network. The positive effect of an immunotherapy using tumor-associated antigens or tumor cells of ovarian carcinomas has not yet been proven. Although many unique tumor-associated antigens have been described and a tumor rejection could be seen in occasional cases, the failure of the immune system to destroy tumor cells is not clearly understood. An alternative approach is to initiate the idiotypic network utilizing antibodies (Ab1 or 2) against a tumor-associated antigen, which induces the production of anti-idiotypic-antibodies (Ab2 beta), mimicking the "internal image" of the tumor-associated antigen. These antibodies are able to induce a specific antitumor immunity in two ways: (1) the Ab2 can present the critical epitope in a different way and so modulates the immune system, or (2) it can induce the production of an Ab3, which by itself binds to the tumor antigen. Our first results on 22 patients with advanced ovarian carcinomas show that the induction of an anti-idiotypic antibody (Ab2 beta) against OC 125 mimicking the TAA Class III CA 125 leads to a prolongation of the survival rate also for extended stages. We see a beneficial role of the induction of the idiotypic network against a tumor-associated antigen showing delayed clinical courses of the disease after vaccination of the patients with antibody fragments of the OC 125.


[Main anticancer substances of vegetable origin]. During the last thirty years, the systematic screening of thousands of vegetal extracts has led to the isolation of numerous antitumor agents which belong to various chemical series. This paper only deals with some of them which are either of current clinical use or under advanced clinical experimentation, e.g., ellipticine, homoharringtonine, camptothecine, acronycine and their derivatives. The origin, the biological activity and its mechanism, and the toxicity of each of these alkaloids are described. These examples highlight the interest of the Plant Kingdom as source of biologically active new structures and the importance of a good knowledge of the mechanism of the activity and toxicity of active components. This knowledge gives a rational basis to prepare compounds of increased activity or reduced toxicity and to use them at their best in therapeutic.


[In vivo and in vitro antitumoral effects of an extract from Vibrio cholerae in different murine models. II. Antitumoral effect in vitro of an extract from Vibrio cholerae: inhibition of 3LL cell growth and stimulation of murine splenocytes]. A bacterial extract from Vibrio cholerae, the DGZ, has been separated into two fractions by gel filtration. The effects of these two fractions and that of the crude extract are investigated and compared by two different in vitro tests. It appears that these extracts exert direct inhibition over the growth of 3LL cultured cells and that they trigger splenocytes in vitro proliferation.


[Cell-matrix interactions and odontoblast differentiation]. The terminal differentiation of odontoblasts requires the integrity of the cytoskeleton and is controlled by cell-matrix interactions. These interactions implicate both matrix molecules and matrix-associated growth factors. On the one hand, predentin-dentin constituents were found to initiate odontoblast differentiation and to allow the maintenance of this state; TGF-beta or related molecules are implicated. Fibronectin on the other hand can induce the differentiation of second generation odontoblasts and interacts with three high molecular weight proteins present in membrane prepared from dental mesenchymal cells. One of these proteins (165 kDa) was localized on the surface of odontoblasts and is involved in the organization of microfilaments. Two main axes of research will have to be developed in the future in order to understand how matrix molecules and growth factors interactions can be modulated in time and space by epithelial and mesenchymal cells, and how such modulations can affect the phenotype of these cells.


[Immortalization of cultured cells by the Ha-rasEJ oncogene and expression of a gene of biogenetic value]. The double recombination into NIH-3T3 cells of the cloned Ha-rasEJ oncogene and the cDNA gene of the human growth hormone (hGH) under the control of a heat inducible promoter (hsp70) allowed hGH production either in vitro using the mass culture of engineered cells in biogenerators, or in vivo after xeno-transplantation of the cells into an animal host. Therefore the in vivo synthesized hGH induced the production of anti-hGH polyclonal antibodies.


[Bioluminescence and creation of chimeric cellular models]. The response of a chimeric cellular model to a hormone has to be more specific, more sensitive, more easily measurable and quantifiable than the natural response. Hence, it can be used either in fundamental and applied studies, for example to analyse hormonal interactions, or to screen new hormone molecules. In the field of steroid hormones, we took benefit from the advantages of the firefly luciferase as receptor gene (simplicity and rapidity of the measurement, high sensitivity, handiness ...) to construct such chimaeric models. Such cell lines were obtained by developing another original use of this reporter gene based on an easy detection of the luciferase activity in intact living cells and using a photon counting camera. This was successfully adapted to obtain and to detect stable transfected cells.


[Using examples of firefly luciferase gene for the study of biological activities (estrogens, retinoids, phorbol esters)]. Some possible applications of chimeric cellular models, specifically responding to an effector through firefly luciferase induction are presented with the help of examples in relation with the biological activity of estradiol or retinoic acid, or phorbol ester. A comparison of experiments on either chimeric or natural responses shows that: i) the responses of both type of cellular models are effector concentration-dependent; ii) these concentrations are in the same order of magnitude; partial agonist compounds and antagonist compounds; iv) potencies (EC50) of test-compounds are similarly classified. Moreover we show that a chimeric cellular model allows the observation of interactions between hormone or effector pathways: it allows readily performed kinetic studies and long-term experiments in intact cells that permit to investigate the effect of a given effector, its reversibility and time-dependent action. Therefore, various steps of a cellular signalling pathway involved in the action of an effector may be observed with such a valuable tool.


[Control of gene expression by antisense nucleic acids]. The use of antisense RNA or of antisense oligonucleotides for the specific control of viral or cellular genes expression has undergone rapid developments recently; their respective advantages and drawbacks will be discussed. Progresses in oligonucleotides chemistry have lead to the synthesis of analogs with improved pharmacological properties. Besides the antisense approach, which usually targets translation initiation or splicing sites, it is possible to interfere specifically with gene expression through triple helix formation (anti-gene strategy) or through the titration of regulatory proteins (sense approach). A major problem encountered in the use of synthetic oligonucleotides is their delivery to their nuclear or cytoplasmic targets after cell uptake by an endocytic pathway; our own work in this field will be discussed. Finally, we will describe the strategies followed by our group to improve the bioavailability of antisense oligonucleotides, as for instance conjugation to poly (L-lysine) or encapsidation in antibody-targeted liposomes.


[Transgenesis and gene therapy].

[Transgenesis and gene therapy]. There are now rather straightforward methods to create transgenic animals whose genome is altered at the germline level. One method consists in the micro-injection of a gene into the pronucleus of a fertilized egg, the second one involves an homologous recombination event obtained in embryonic stem cells in culture. Only the latter method could eventually lead to an authentic gene therapy since it could actually substitute a normal gene for a mutated one instead of merely introducing a supplementary gene as done by micro-injection. Description of these techniques makes it obvious that germline therapy of human beings would not only be inacceptable on ethical grounds but would also hardly have any medical indications. Quite on the contrary, somatic gene therapy does not suffer from the same reservations and has numerous potential applications to man. As a matter of fact, several protocols have already received approval and have reached the stage of clinical trials: for example SCID (severe combined immunodeficiency due to a mutated adenosine deaminase gene), AIDS as well as some forms of malignant tumors.


Deregulated c-fos augments the expression of IL-2 gene in T cells stimulated with anti-CD3 antibody. The expression of IL-2 gene in T cells stimulated with immobilized anti-CD3 antibody is controlled by several transcription factors like as AP-1, IL-2 kappa B and NFAT. We studied an effect of deregulated c-fos, a component of AP-1, in the expression of IL-2 gene in T cells by using splenic T cells from transgenic mice carrying the exogenous c-fos gene under the control of the H2-Kb promoter (H2-c-fos). The IL-2 production of H2-c-fos T cells stimulated with anti-CD3 antibody was enhanced and prolonged. Levels of AP-1 and IL-2 kappa B transcription factors in the nuclear extract from the H2-c-fos T cells were also augmented. These results suggest that c-Fos is one of the major regulatory factors for the expression of IL-2 gene in T cells activated with anti-CD3 antibody.


Effect of intra-aortic balloon pumping after cardiac surgery: comparative study of use in coronary bypass and valve replacement. To compare the effects of intra-aortic balloon pumping (IABP) between use for coronary artery bypass grafting (CABG) and for valve replacement, the results were reviewed in 513 patients who underwent CABG (n = 215) or valve replacement (n = 298). The rate of weaning from IABP and in-hospital survival after IABP support were excellent in CABG patients, where all 21 CABG patients (100%) and 17 out of 25 valve replacement patients (68%) could be weaned from IABP, and 19 out of 21 CABG patients (90%) and 14 out of 25 valve replacement patients (56%) survived for a long period. From comparative studies of pre- and postoperative data, the duration of IABP support in patients who could be weaned from IABP or who achieved in-hospital survival tended to be more prolonged in patients with valve replacement than in those with CABG. Among patients having IABP postoperatively, 4 CABG patients and 8 valve replacement patients had definite indications for IABP support. In the other 34 patients, preoperatively depressed cardiac function was observed in 12 patients where most of them were patients with valve replacement. Postoperative improvements in cardiac index were not as marked in patients undergoing valve replacement with IABP. The outcome for patients who had definite causes of post-operative cardiac failure, which were more numerous in the valve replacement group, was poor even if they could be weaned from much longer IABP support. In conclusion, the IABP was more effective in patients with CABG compared with those with valve replacement, because the number of patients who had a depressed cardiac function during the pre- and postoperative periods was lower in the CABG group and the incidence of serious perioperative complications was less frequent in these patients.


Ultrastructural studies of the bile duct in alcoholic liver disease. The fine structural characteristics of the bile duct in patients with alcoholic disease are described. Dark cell metamorphosis, edematous microvilli, and increased number of pinocytotic vesicles on the basal wall surface of the duct epithelium were observed. These alterations may be interpreted as evidence of disordered water metabolism, probably reflecting secretion and reabsorption hyperfunction in the duct epithelium. In addition, widened intercellular spaces in the basal half of the epithelium suggested retention of fluid following reverse pinocytosis along the lateral cell surface. Although no alterations of the duct epithelium distinct from those in patients with other liver diseases were apparent in patients with alcoholic liver disease, the basement membrane of the bile duct exhibited unusual duplication with multiple layers and occasional loop-formation in lacunae on the basal surface.


Effects of angiotensin II on regional brain noradrenaline metabolism in non-stressed and stressed rats. By measuring levels of noradrenaline (NA) and its major metabolite, 3-methoxy-4-hydroxyphenylethyleneglycol sulfate (MHPG-SO4), in various rat brain regions, the effects of angiotensin II (AT II) injected intracerebroventricularly (ICV) were investigated in non-stressed and stressed rats. Immobilization stress with a wire mesh was employed. Immobilization stress induced significant increases in MHPG-SO4 levels in all brain regions examined, i.e., hypothalamus, amygdala, thalamus, midbrain, hippocampus, cerebral cortex and locus coeruleus (LC) region, which suggests that stress increases NA release in these regions. AT II (1 and 10 micrograms), injected ICV 5 min before exposure to stress for 60 min, attenuated stress-induced increases in MHPG-SO4 levels in the hypothalamus, thalamus and amygdala. AT II, injected ICV in non-stressed rats at 65 min before sacrifice, significantly elevated MHPG-SO4 levels in the midbrain, cerebral cortex, thalamus and LC region. These results indicate that the attenuating effect of AT II on stress-induced increases in brain NA release are, in part, related to the reduction of fear and/or anxiety of the animals exposed to stress.


Myocardial ischemia in Kawasaki disease; evaluation by dipyridamole stress thallium-201 (Tl-201) myocardial imaging and exercise stress test. The detection of myocardial ischemia is essential for evaluation of patients with Kawasaki disease, especially who have had coronary artery lesions. To evaluate the clinical efficacy of Tl-201 single photon emission computed tomography (SPECT) after dipyridamole infusion (maximum dose 0.70 mg/kg) for detecting myocardial ischemia, 44 patients with Kawasaki disease aged 7.7 +/- 4.8 years at the study and 10 age matched controls were observed. In the Kawasaki disease group, significant coronary artery stenosis was observed in 14, coronary aneurysm without stenosis in 18, the regression of the coronary aneurysms in 2 and without coronary lesions in 10 patients. In 22 of 44 patients, treadmill exercise stress test was also performed at the same period. Myocardial ischemic changes were observed in 11 patients, all combined with significant coronary artery stenosis. The sensitivity of SPECT for detection of overall coronary stenosis was 79%, contrary that of treadmill exercise test was only 33% (p < 0.001). Furthermore, among the patients having significant coronary stenosis, the severity score was significantly elevated in patients who had electrocardiographic abnormal Q wave compared to those without abnormal Q wave (51.0 +/- 38.8 versus 20.0 +/- 12.1, p < 0.05). These data suggest that the pharmacological stress scintigraphy using dipyridamole injection provides not only the accurate detection but quantitative evaluation of myocardial ischemia in these patients. This noninvasive technique may become one of the most useful index for detection and follow-up of myocardial ischemia in Kawasaki disease.


Concentrations of interleukin-1 beta, interleukin-6, interleukin-8 and TNF-alpha in cerebrospinal fluid from children with septic or aseptic meningitis. Cytokines at an inflammatory site may be a better indicator of the clinical severity of an infectious disease than the serum levels of the cytokines. Concentrations of interleukin-1 beta (IL-1 beta) in paired samples of cerebrospinal fluid (CSF) from 10 rabbits with experimental bacterial meningitis caused by H. influenzae type b, were measured, and compared to the concentrations of four cytokines; IL-1 beta, interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) in CSF samples from 45 children with or without meningitis. The IL-1 beta concentrations in the CSF from rabbits with experimental meningitis were significantly higher than the concentrations in control animals without meningitis (p < 0.001). The mean CSF concentrations of IL-8 from meningitic children were significantly higher than in the control group without meningitis (p < 0.005). TNF-alpha was only detected in septic meningitis. Assays of IL-6, however, were not significantly different in the septic meningitis group, the aseptic meningitis group and the non-meningitis group. These data indicate a possible role of IL-1 beta, IL-8 and TNF-alpha as mediators in the meningeal inflammatory process in patients with meningitis and TNF-alpha, in particular, may play a role in the pathogenesis of septic meningitis.


Clinical significance of thallium-201 single-photon emission computerized tomography (Tl-201 SPECT) in the evaluation of viability of gliomas. The clinical significance of Thallium-201 single-photon emission computerized tomography (Tl-201 SPECT) in the evaluation of viability of gliomas was studied comparatively with histological examination of tumor tissue using Ki-67 and proliferative cell nuclear antigen (PCNA) monoclonal antibody. The relationship between radionuclide uptake of Tl-201 in tumor specimens and labeling indices of special staining using Ki-67 and PCNA monoclonal antibodies were also studied. The population studied consisted of 17 patients with glioma. Tl-201 indices obtained from early and delayed images and its washout rates were used for quantitative analysis of Tl-201 SPECT findings. Tl-201 indices showed high values according to the histological malignancy of gliomas. Radionuclide uptake of Tl-201 in the tumor specimens were also high in those with high labeling indices of Ki-67 and PCNA. The lesions with marked Tl-201 uptake on early and delayed images had numerous Ki-67 and PCNA positive-stained cells. The lesion with low Tl-201 washout rate therefore reflected well the more viable lesion in the tumor tissue. It is concluded that the viability of gliomas including anaplastic changes and response of gliomas to the treatment can be detected by serial study with Tl-201 SPECT.


Comparison of incidence, mortality and treatment of acute myocardial infarction in hospitals in Japan and China. Acute myocardial infarction (AMI) is one of the most intractable diseases and is increasing rapidly in Japan and China. Two hospitals in Japan and China, the Critical Care Center of Kurume University Hospital and the Chinese Beijing 309 Hospital in China (abbreviated to Beijing 309 Hospital) were compared. The incidence, mortality and treatment of AMI were investigated in both hospitals from 1989 to 1991. The incidence of AMI for all patients admitted during the three years was 5% in Kurume University Hospital and 4.7% in Beijing 309 Hospital, which are similar rates. The average age of the patients in Beijing 309 Hospital was younger (58 +/- 13) than in Kurume University Hospital (64 +/- 11). The mortality rate in Kurume University Hospital was slightly lower than the rate in Beijing 309 Hospital (8.1% vs 8.9%). Thrombolytic therapy was actively performed in both hospitals. In Kurume University Hospital, urokinase (UK: 71.4%) or recombinant tissue plasminogen activator (rt-PA: 28.6%) was administered by intravenous (85.7%) and intracoronary percutaneous transluminal coronary recanalization (PTCR: 14.3%) injection. In Beijing 309 Hospital, UK (32.7%) or snake poison enzyme (SPE: 62.3%) was administered by intravenous (85.8%) or intra-aortic (14.2%) injection. Rt-PA was only used in Japan and SPE was only used in China, but both had very strong fibrinolytic effects and resulted in high success rates of coronary reperfusion. The incidence of direct coronary intervention with percutaneous transluminal coronary angioplasty (PTCA) and intra-aortic balloon pumping (IABP) for cardiogenic shock was much higher at Kurume University Hospital than at Beijing 309 Hospital.(ABSTRACT TRUNCATED AT 250 WORDS)


A study of the role of sex hormones in rat ovarian cancer. The effects of estrogen, progesterone, and testosterone on the growth of 7,12 dimethylbenz(a)anthracene (DMBA) induced adenocarcinomas in rats (Wistar strain) were evaluated. Estrogen resulted in the highest acceleration of tumor volume. The histologic features were a solid structure associated with a significant proliferation of connective tissues and with many signet ring cells with intracytoplasmic canaliculi. Progesterone changed the histologic features to a more immature adenocarcinoma associated with a notable solid area with many mitotic figures, although the growth rate of the tumor was the same as the controls. On the contrary, testosterone induced the slowest tumor growth and a histologically scirrhus pattern. The results of this preliminary observation indicate a possible role for sex steroids in the ovarian tumorigenic process.


Non-invasive evaluation of pulmonary arterial and right ventricular pressures with contrast enhanced Doppler signals of tricuspid regurgitation flow using sonicated albumin solution. To determine the feasibility of the non-invasive determination of systolic pressure of the pulmonary artery and the right ventricle in pediatric patients, the velocity of tricuspid regurgitation was measured in 30 patients using a contrast enhanced Doppler echocardiography. After sonicated albumin injection, trivial tricuspid regurgitation signals were enhanced in 27 patients (90%). Peak systolic velocity was not altered by before and after sonicated albumin injection in 2 patients. Right ventricular (RV) systolic pressure obtained by continuous wave Doppler during sonicated albumin enhancement corresponded very closely to that measured by catheter in 27 patients (r = 0.96). In 27 patients, difference of estimation of RV systolic pressure by non-enhanced Doppler and enhanced Doppler with sonicated albumin was statistically significant (32.3 +/- 27.6 mmHg versus 2.9 +/- 7.7 mmHg p < 0.001). Systolic pressure of pulmonary artery was estimated by RV systolic pressure measurement (by enhanced Doppler method) minus peak pressure gradient across the pulmonary valve (non-enhanced Doppler method). Pulmonary arterial systolic pressure measured by enhanced Doppler method and that by catheter method were highly significant (sonicated albumin method, r = 0.95). This technique may be a valuable non-invasive method for determining an accurate right ventricular and pulmonary arterial systolic pressures in this setting.


[Strains of Salmonella with atypical biochemical activity found in diagnostic material]. Biochemically atypical strains belonging to 11 genera of Salmonella subspecies I in the number of 200 were collected in Poland in the years 1980-1991 were tested by means of 22 biochemical tests. Atypical property stability was detected in 164 strains. Most stable properties were following: ability to ferment lactose, lack of ability of dulcitol fermentation, lack of hydrosulfide production and lack of lysine decarboxylation. Atypical properties were occurring singularly or clusters numbering two or three. Identification procedure for Salmonella accepted in this country enables proper diagnosis of an atypical strain, however it requires application of supplemental biochemical tests. These strains were tested for susceptibility of 15 antimicrobial agents and no correlation between antibiotic resistance and occurrence of one or several atypical properties has been found.


[A trial application of the latex test for evaluation of antibody level in rabbit immune sera]. A latex test was elaborated which served for evaluation of quality of rabbit immune sera for antigen 0 of selected Gram-negative bacteria. Sensitivity and specificity of this test in comparison with passive hemagglutination and immunoenzymatic DOT-ELISA reactions was evaluated. These studies were performed on immune sera for antigen O of Salmonella groups B, C1, C2, D and E, Yersinia pseudotuberculosis and in antigen preparations for above listed microorganisms both in homologous and heterologous systems. It was found that sensitivity of the latex test is 9 to 160 times lower than that of passive hemagglutination and 7 to 307 lower than for DOT-ELISA. Sensitivity of the latex test and passive hemagglutination reaction was evaluated on the basis of results of cross reaction between studied antigens and unabsorbed rabbit sera, establishing so called sensitivity indexes, which were informing how many times heterologous titer is lower than homologous titer. So evaluated sensitivity of the latex test was close to sensitivity of the passive hemagglutination reaction. It was found that slide latex test is characterized by satisfactory sensitivity and good sensitiveness and may be used for evaluation of antibody level 0 antigens of Salmonella and Yersinia. The value of this test is characterized by high repeatability of results, as well as low work and time-consuming.


[Investigation of factors determining stability of BCG vaccine]. This study was aimed at establishment of influence of a carrier-sodium glutamate and of changes introduced during the process of freeze-drying on survival of BCG bacilli during lyophilization, as well on thermostability and homogeneity of the vaccine and its immunogenicity. It was found that appropriate drying of the vaccine after freeze-drying performed in higher temperature influences favorably its thermostability. Concentration of the carrier is significantly influential for survival of bacilli during freeze-drying. Vaccine containing 1% of sodium glutamate was characterized by best thermostability, homogeneity and high survival of bacilli during freeze-drying. These parameters were keeping on a high level also one year after expiration date. It was shown that there exists a proportional dependence between immunogenicity of the vaccine measured indirectly by intensiveness of tuberculin allergization and number of live particles of BCG contained in vaccination dose.


[Dependence between penetration route and course of infection with Coxiella burnetii in mice]. This study was aimed at investigation of course of Coxiella burnetii infection in mice infected by these bacteria by different routes. The animals infected intranasally, perorally, intraperitoneally and intravaginally by suspension of C. burnetii cells. Mice were also infected via peritoneal and intravaginal route with spermatozoa derived from infected males. In all animals at the same time specific antibodies against phase I and phase II antigens of C. burnetii belonging to IgG and IgM classes of similar titers appeared and this was detected by dot-blot immunoenzymatic test. Independently of route of infection C. burnetii were present in the liver, spleen, testicles, prostate and spermatozoa of tested animals. The bacteria were detected in these organs for 18 days of infection, in the blood for 7 days only, whereas in urine they appeared as late as 14 days after infection. The course of infection with C. burnetii in mice in thus similar regardless of site of bacterial penetration. Infection with C. burnetii may be also transmitted by a sexual route from male to female animals. Infection of female mice occurs both after intravaginal application of live suspension of C. burnetii or spermatozoa derived from infected males.


[Occurrence of infections with C burnetii (Q fever) in persons from increased risk groups in Poland--serological survey and evaluation of studied methods]. During years 1988-1991 at Regional Sanitary and Epidemiological Stations in Białystok, Kielce, Lublin, Leszno, Piotrków Trybunalski, Lódź, Poznań, Plock, Opole, Sanok, Skierniewice and Zielona Góra--serological studies in persons amenable to compulsory check-up against brucellosis were performed for detection of infections with C. burnetii. In whole, 20651 persons were investigated on the Polish territory and it was found that in 22.8% antibodies against antigens of C. burnetii are present. Percentage of persons with detected antibodies varied in different regions from 0 to 41.7%. No antibodies were found in persons inhabiting regions of Kielce, Piotrków Trybunalski and Skierniewice. Most persons with antibodies indicating contact with C. burnetii were found in West Poland, namely in the Leszno and Poznań region. As in diagnosis of Q fever two methods (OWD and OMA) or OWD only were applied, it is difficult to compare results obtained in regions using two or only one method of determination.


[Evaluation of usefulness of OWD and OIEP in diagnosis of epidemically occurring infections caused by Mycoplasma pneumoniae]. Blood serum samples of 3593 persons clinically suspected of infection with Mycoplasma pneumoniae were tested. Of these, patients with pneumoniae constituted 66.5%, upper respiratory tract infection--24.0% and with symptoms localized outside the respiratory system--9.5%. These studies were performed by application of complement fixation test (OWD) and immunoelectroprecipitation (OIEP) methods, accepting as a diagnostically significant--titer 1:60 or higher and/or occurrence in OIEP reaction with serum diluted 1:2 or more. Among patients studied prevailed children in the age of 3 to 16 years (61.6%). Mycoplasmosis was detected in these patients in 1071 out of 2236 cases (47.9%). Compatible results in both tests were obtained in 90.6% patients, whereas OWD only in 3.0% and OIEP only in 6.4% cases. Simultaneous application of both tests increased detectability of infections caused by M. pneumoniae by 3% in relation to OIEP and by 6.4% in relation to OWD.


[Influence of high dose intravenous immunoglobulins (IVIG) on HSV-1 infection in immunosuppressed mice]. Influence of low and high doses of intravenous immunoglobulin (IVIG) was compared on the basis of survival studies of mice infected with temperature sensitive (ts 28 degrees C) clone of Herpes simplex virus (HSV-1) after previous treatment with cyclophosphamide (CY), Sandoglobulin containing intact IgG and Gamma-Venin containing only fragments of (Fab)2 IgG were used. It was found that 4 and 16 mg doses protected animals in contrast to the 32 mg dose of Sandoglobulin which protected mice in only small degree.


[Changes in the level of humoral response for measles virus in patients with subacute sclerotic panencephalitis during 1984-1989]. Immunological response to measles virus in serum and cerebro-spinal fluid of 338 patients with subacute sclerotic panencephalitis (SSPE) during the years 1984-1989, was investigated. It was found that mean titer and distribution of results of antibody determination by the hemagglutination inhibition (OZHA) method in patients sera are visibly differing during consecutive years of investigation. Analysis of these parameters in determination by the immunoenzymatic methods (ELISA) has shown their higher stability. Also results of determinations of IgG concentration in sera of patients with SSPE did not demonstrate significant differences between consecutive years of investigation. Systematic decrease of mutual relation between antibodies to measles virus detected by ELISA and OZHA was found. Analysis of local synthesis of antibodies to measles virus in the central nervous system has revealed that it most strongly expressed by hemagglutinin.


[Use of western blotting methods in diagnosis of viral infection causing tick-borne encephalitis]. A Western blot method for investigation of antigens and of immunological response to tick-borne encephalitis was prepared. An analysis was performed testing cross reactions of proteins of prepared antigen with sera for four Polish strains of tick-borne encephalitis and four other flaviviruses. Usefulness for diagnosis of infections in humans was determined by comparison of Western blot method with ELISA immunoenzymatic test. It was found that elaborated method permits equally as ELISA for evaluation of immune response in immunoglobulin classes. By evaluation of response for individual virus components, Western blot method enables at the same time verifications of nonspecific determinations which result from reaction of antibodies present in tested diagnostic materials with nonviral antigens.


[Evaluation of contents of A and B group substances in biological preparations]. The study was aimed at determination of blood A and B group substances in biological preparations used in Poland. Twenty three series were investigated, namely: Di-Te-Per, Ty-Te, Ty, Te, against cholera, vaccine according to Delbet and Panodin. Also were tested: 65 series of imported preparations of immunoglobulin g (i.v.) such as Endobulin, Sandoglobulin, Gamma-Venin, Veinoglobuline and 5 local series such as Bioglobulin, as well as 9 series of preparation LNI (i.m.) Human Gamma Globulin. Presence of substance A was detected in tetanus and botulinum horse antitoxins in amount from 3.75 micrograms/ml to 30 micrograms/ml. Group substances A and B contained 6 series of LNI preparations-Veinoglobuline. Amount of substance A was detected as 3.75 micrograms/ml-7.5 micrograms/ml and of substance B as 2,5 micrograms/ml-5 micrograms/ml. Group substances A and B were not present in vaccines used according to vaccination calendar.


[Investigation of immunogenic properties of diphtheria antitoxin by intradermal active test of potency]. Usefulness of four variants of intradermal test for routine testing of protective potency and diphtheria anatoxin was studied. Comparing with classical active test it was found that full intradermal test possesses data system and results comparable to the classic test and may replace it. This in effect results in four-fold reduction of guinea pig number necessary for performance of the investigation. As orientating test, estimate determining AnaDi potency a single dose performed on six guinea pigs was selected. In the intradermal test the time of investigation, as compared with the classic test, may be shortened by one week.


[Simplified active test of estimating tetanus toxoid]. The main purpose of these investigations was the comparison of two potency tests of Tetanus Toxoid. It was found that two doses test with reduced number of animals can be used in assay of different vaccines containing Tetanus component. One dose test is agreeable with requirements of WHO. It is credible and not complicated method of estimation Diphtheria and Tetanus Toxoids. This test required minimal number of animals.


[Investigation of activity of new quaternary imidazole-linear chlorides against selected bacterial strains. Synthesis of 2-alkyl-3-alkymethyl-1-ethylimidazolelinear and 2-alkyl-3-alkoxymethyl-1-ethylimidazolelinear chlorides]. Synthesis and bacteriostatic properties of a new alkylthiomethylimidazole-linear and alkomethyly-limidazole-linear chlorides was described. It was found that these chlorides are active against bacilli, cocci and yeast-like fungi and express lower activity against rods. Highest activity was observed with 2-benzyl-3-decyloxymethyl-1--ethylimidazolinear and 1-ethyl-heptadecyl-3-undecyloxymethylimidazolelinear chlorides.


[Evaluation of susceptibility to antibiotics of Staphylococcus aureus strains resistant to methicillin]. Methicillin-resistant strains of Staphylococcus aureus (MRSA) constitute a serious diagnostic and therapeutic problem. Over 500 strains of Staphylococcus aureus were tested for susceptibility to methicillin. By application of a screening method, 13.7% of these strains were classified as methicillin-resistant. Over 95% of these strains were isolated from hospital infections. Applying criteria of belonging of these strains to methicillin-resistance classes it was found that 49.3% belonged to class II, 31.2% to class III and 19.5% to class IV. Analysis of susceptibility to antibiotics of MRSA strains demonstrated significant differences between class II and between class III and IV in resistance to imipenem, gentamycin, erythromycin and tetracycline. All tested strains were susceptible to ciprofloxacin, ofloxacin, vancomycin and teicoplanin. The screening method (25 mg methicillin/l of TSA medium) results in obtaining of univocal results of determination of methicillin-resistance in S. aureus.


Changes in the rat brain 5-HT1A and 5-HT2 receptors after chronic administration of levoprotiline, (+)-oxaprotiline and other antidepressant drugs. The effects of levoprotiline (LEV), a (-)-enantiomer of oxaprotiline (OXA) and a clinically effective antidepressant, on the binding parameters of hippocampal 5-HT1A and cortical 5-HT2 receptors of rats were compared with those of (+)-enantiomer of OXA ((+)-OXA), imipramine and mianserin. Both LEV and (+)-OXA displayed in vitro some affinity for 5-HT1A receptors labelled with [3H]-8-OH-DPAT, and for 5-HT2 receptors labelled with [3H]-ketanserin. Repeated administration of LEV, for 14 days led to a marked increase in the number of 5-HT1A binding sites in the rat hippocampus, with no change in the KD values. (+)-OXA, imipramine and mianserin produced similar effects on 5-HT1A binding parameters. The number of 5-HT2 receptors was increased after two weeks of LEV administration, not altered after (+)-OXA, and decreased after imipramine or mianserin. The number of [3H]-ketanserin binding sites was decreased after four weeks of (+)-OXA administration, but not altered after LEV. The specific binding of [3H]-ketanserin in the rat cerebral cortex was decreased after repeated treatment with LEV and (+)-OXA (ex vivo). In competition studies the affinity of serotonin for [3H]-ketanserin binding sites was decreased in LEV- and increased in (+)-OXA-treated rats. The results suggest that LEV similarly to other antidepressants increases the number of 5-HT1A receptors, however without common alteration in 5-HT2 receptor number and function.


[Effect of different vascular prosthesis materials on blood platelets, blood clotting and fibrinolysis]. Studies were performed on Dacron vascular prostheses (USA), Gore-Tex (Germany), polyester prostheses (Czechoslovakia), Lawsan (Russia), as well as on DALLON-standard and DALLON-double velour prostheses (Poland), non-impregnated or impregnated with collagen, with heparin and albumin, with antithrombotic and antibacterial preparation, and with antibacterial preparation. In vitro, all the materials of vascular prostheses cause an adhesion of platelets in different degree, increase the availability of platelet factor 3, release platelet factor 4, reduce the blood clot retraction, shorten clotting time of the whole blood as well as recalcination time of intact plasma and increase the prothrombin consumption. None of the materials induced platelet aggregation nor influenced the activity of fibrinolytic system. Prostheses impregnated with collagen or with heparin and albumin were the most thrombogenic. Those impregnated with antibacterial preparation or with antibacterial and antithrombotic preparations, and Gore-Tex prostheses showed the lowest thrombogenicity. Thrombogenicity of vascular prostheses not only depends on chemical structure of the material, but on the method of fibre manufacturing by individual producers as well.


Structure-activity relationship studies of CNS agents. Part VIII. Bulk tolerance around the protonation center of 4-substituted 1-(3-chlorophenyl)piperazines at 5-HT1A and 5-HT2 receptors. The effect of a steric hindrance around the protonation center of the model 4-substituted 1-(3-chlorophenyl)-piperazines 1-9 and 11-14 on their affinity for 5-HT1A and 5-HT2 receptor sites was investigated. Additional evidence for hydrophobic interactions between the N-4 hydrocarbon substituents and 5-HT1A receptors has been presented. However, the hydrophobic forces play a minor role in stabilization of the bioactive complex with 5-HT2 receptors. It has also been found that even bulky substituents around the protonation center of 1-aryl-piperazines are well tolerated at both 5-HT1A and 5-HT2 sites.


[Changes in the activity of the clotting system and fibrinolysis components in the layers of double velour DALLON grafts at different periods after implantation into the aorta]. During one year experiment, the thromboplastic, antiheparin, plasminogen activator, antithrombin and antiplasmin activities were evaluated in the homogenates of the intimal, medial and adventitial layers of polyester double velour DALLON prostheses implanted into the dog abdominal aorta. It was found, that 7 days after implantation the thromboplastic, antiheparin and plasminogen activator activities in DALLON graft neointima were high, whereas those of the antiplasmin and antithrombin were low. These changes facilitate the sealing of the prosthesis pores and at the same time prevents thrombosis. Four months after implanting, the activity of hemostatically active components in the various graft layers became similar to the activity of aorta components. Rapid decrease of the thrombogenic potential and increase of the fibrinolytic activity in layers of double velour DALLON grafts facilitate the maintenance of graft patency.


Synthesis, physicochemical and preliminary pharmacological properties of N-[beta-hydroxy-gamma-(N-phenylpiperazinepropyl)]-2-pyrrolidinone. The present paper reports on the synthesis and preliminary pharmacological properties of N-[beta-hydroxy-gamma-(N-phenylpiperazinepropyl)]-2- pyrrolidinone (MG-1). MG-1 was obtained by aminolysis of 1-(beta, gamma-epoxypropyl)-2-pyrrolidinone and N-phenylpiperazine. Its structure was established by elemental and spectral analyses (IR, UV, MS, 1H, 13C, 2D H-H and 2D C-H NMR). The antiarrhythmic activity of MG-1 was investigated on mice, rats and guinea pigs, using several models of arrhythmia. MG-1 attenuated or prevented the adrenaline- and barium chloride-induced arrhythmia. MG-1 demonstrated potent local anesthetic properties and depressed the depolarization phase of the action potential of cardiac cells. These results indicate that MG-1 possesses antiarrhythmic activity.


[Comparison of cathepsin D activity of DALLON-standard and DALLON-double velour grafts implanted into the abdominal aorta]. Cathepsin D activity was compared during the healing period of polyester DALLON-standard and DALLON-double velour prostheses implanted into the abdominal aorta of dogs. During first months following the implantation cathepsin D activity is considerably higher in all the layers of both types of prostheses than in the aorta. The normalization of this enzyme activity occurs significantly faster during the healing of DALLON-double velour prostheses than in that of DALLON-standard. This fact shows greater progress in the protein metabolism normalization during the process of healing and tissue reconstruction in DALLON-double velour prostheses.


[Comparison of healing characteristics of DALLON-standard and DALLON-double velour vascular prostheses]. By means of histological method and the scanning and transmission electron microscope healing processes of two types of Polish prostheses: DALLON-standard and DALLON-double velour were compared. The experiments were carried out on 36 dogs. The two types of prostheses mentioned were implanted in abdominal aorta. The grafts were removed 1, 4 and 12 months after implantation. The performed studies showed that although the sequence and nature of morphological changes leading to the reconstruction of the vascular wall in both types of grafts were similar, the development of the neointima as well as the formation of the neo-adventitia firmly accreted with surrounding tissues were considerably faster in double velour DALLON prostheses. Unlike in standard DALLON prostheses, the neointima of double velour DALLON grafts was rich in cells, had elastic fibres. Already after 12 months double velour DALLON grafts were completely covered by endothelial cells, which proves better reconstruction of the vascular wall. Rapid endothelial cells coverage of the lumen surface prevents thrombosis. Morphological differences observed in endothelial cells covering the neointima of standard grafts and double velour graft suggest the existance of various sources of endothelialization in both types of grafts.


The role of GABA-ergic signal in the regulation of melatonin biosynthesis in vertebrate retina. The in vivo effects of GABA-ergic drugs on the activity of serotonin N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT), two enzymes involved in melatonin biosynthesis, were investigated in light-exposed chicken retina. The ip administration of muscimol and baclofen (direct agonists of GABA-A and GABA-B receptors, respectively), aminooxyacetic acid (an inhibitor of GABA transaminase), and nipecotic acid (an inhibitor of GABA reuptake), significantly increased the retinal NAT activity by 50-100%. Similar rises in NAT activity were observed following intraocular treatment of ether-anesthetized chickens with muscimol, baclofen and GABA. In contrast to NAT, there was no effect of the tested drugs on the retinal HIOMT activity. Aminophylline (a phosphodiesterase inhibitor) markedly elevated the retinal NAT activity, and a combined treatment with the GABA-ergic drugs and aminophylline resulted in additive effects. The actions of both muscimol and baclofen were antagonized by picrotoxin and bicuculline (two GABA-A receptor blockers), whereas the effect of baclofen was not changed by a selective GABA-B receptor blocker, CGP 35,348. Melatonin given ip significantly raised NAT activity, and its combination with muscimol further stimulated the enzyme. Picrotoxin and bicuculline given to chickens during the dark phase of 12 h light--12 h dark illumination cycle significantly suppressed the nocturnal NAT activity in retina. Neither GABA nor muscimol and baclofen significantly affected basal and forskolin (1 microM)-stimulated adenylate cyclase activity in vitro in light-exposed chicken retina. It is concluded that a GABA signal (acting through type A of GABA receptors) plays an important role in a complex mechanism regulating the rhythmic melatonin biosynthesis in vertebrate retina.


Propranolol analogs containing natural monoterpene structures: synthesis and pharmacological properties. A few derivatives of natural, bicyclic monoterpenes, which are propranolol analogs, were synthetized. Those compounds were studied pharmacologically in order to determine their toxicity, antiarrhythmic activity in selected experimental models of arrhythmia, the local anesthetic effect and influence on the cardiovascular system. The tested compounds showed a less potent or similar toxicity towards reference drugs, were devoid of an antiarrhythmic activity in the model of barium arrhythmia, yet some of them (compounds 9 and 12) increased the arrhythmogenic dose of strophanthin. All the compounds studied had a local anesthetic effect stronger than lidocaine in infiltration anesthesia, and compound 8--also in surface anesthesia.