[Low doses of azidothymidine in the treatment of HIV-1 virus infection].

A group of 26 patients (18 males and 8 females) infected with HIV (42% through sexual route and 58% through blood/blood products transfusion) was prospectively studied to assess the efficacy of low doses (300 mg/day) of AZT combined (n = 15) or not (n = 11) with ACV (600 mg/day). According to CDC stages, 12% were in stage II, 73% in stage III and 15% in stage IV. Patients were followed for a maximum of 156 weeks. An objective response was observed in all patients who improved significantly in: performance status (Karnofsky 74.5 versus 97.6%, p less than 0.01), weight 58.9 versus 68.6 kg, p less than 0.01), and absolute CD4 T cell count (329/microL versus 480/microL, p less than 0.01). The levels of hemoglobin dropped after treatment (12.8 versus 11.5, p less than 0.01). Median survival was 114 weeks for all the group. With the exception of granulocytopenia in 42% of patients treated with AZT + ACV versus only in 22% of those treated solely with AZT (p = 0.02), similar effects were recorded in both treatments: 114-week survival was 60% for those treated solely with AZT, whereas 156-week survival was 93% for those treated with AZT + ACV (p NS), but the response was better for the combination of antivirals in the group of patients with more than 200 CD4 cells/microL at diagnosis as compared with those with less than 200 cells (110-week survival of 100% versus 50% respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Disposition of mesalazine from mesalazine-delivering drugs in patients with inflammatory bowel disease, with and without diarrhoea.

The disposition of mesalazine from the azo compounds sulphasalazine and olsalazine (Dipentum) and from the slow-release mesalazine drugs Pentasa, Asacol, and Salofalk was studied in 20 patients with inflammatory bowel disease. Ten of them had diarrhoea, and 10 had normal stools. On the last 2 days of a 7-day maintenance treatment with each of the study drugs urine and faeces were collected for determination of mesalazine, acetyl-mesalazine, and unsplit azo compound. In patients with and without diarrhoea the urinary and the faecal excretion of acetyl-mesalazine was lowest during treatment with olsalazine. The proportion of acetyl-mesalazine in faeces was highest during treatment with Pentasa in both groups. The presence of diarrhoea was associated with a decrease in the proportion of acetyl-mesalazine in faeces during treatment with all drugs, not significant only for Pentasa. The proportion of unsplit azo compound in faeces increased in the case of diarrhoea to almost 50%. It is concluded that in patients with inflammatory bowel disease diarrhoea substantially influences the disposition from all these drugs except Pentasa.


Clinical evaluation of pancreatic cancer-associated mucin expressing CA19-9, CA50, Span-1, sialyl SSEA-1, and Dupan-2.

We have previously described the purification and partial characterization of a new pancreatic cancer-associated antigen, a pancreatic cancer-associated mucin expressing CA19-9, CA50, Span-1, sialyl SSEA-1, and Dupan-2. This study describes the clinical evaluation of various assay systems for this antigen which depend on measuring respective serum levels. Elevated levels of antigen were detected in the sera from both patients with malignant and non-malignant diseases. However, elevated serum levels of CA19-9 and Lewisa and Lewisb epitopes on moieties were restricted to pancreatic and biliary tract cancers, although adequate sensitivity was not attained. Coordinate evaluation of these three markers improved the sensitivity to some extent without loss of specificity for the diagnosis of pancreatic and biliary tract cancers, because of the heterogeneity of the coexpression of these epitopes. We developed additional assay systems with a combination of this antigen and two lectins (Bauhinia purpurea (BPA) and Vicia villosa (VVA)). Elevated levels of BPA- and VVA-reactive antigens were detected in 41% and 31%, respectively, of pancreatic cancer sera samples. Few patients with chronic pancreatitis had an elevated serum level of either antigen, and higher elevated levels of these markers were restricted to the sera of patients with malignancies. Our results suggest that this antigen is found in the sera of patients with various conditions and in the sera of normal subjects but that antigens bearing CA19-9 or Lewisa or Lewisb epitopes and an altered carbohydrate structure recognized by BPA and VVA lectins are preferentially present in the sera of patients with pancreatic and other malignancies.


Vagal control of the release of somatostatin, vasoactive intestinal polypeptide, gastrin-releasing peptide, and HCl from porcine non-antral stomach.

We studied the secretion of somatostatin and HCl and the release of vasoactive intestinal polypeptide (VIP) and gastrin-releasing peptide (GRP) from isolated, vascularly perfused, porcine non-antral stomach. Electric vagus stimulation increased acid secretion and the release of VIP and GRP and inhibited somatostatin secretion as determined in the venous effluent. Atropine abolished the HCl response and reversed the somatostatin inhibition to a three-fold increase, whereas GRP and VIP responses were unchanged. Both intra-arterial carbachol (10(-6) M) and GRP (10(-8) M) increased acid secretion and inhibited somatostatin secretion. VIP (10(-8) M) increased somatostatin secretion and had no effect on acid secretion. By immunohistochemistry, somatostatin was localized to both open-type and closed-type cells equally spread in the various parts of the gastric glands without particular relation to the parietal cells. Numerous GRP- and VIP-immunoreactive nerve fibers were seen between the glands. It is concluded that the fundic and antral secretion of somatostatin, investigated in a previous study, are differently regulated. The relation of fundic somatostatin release to acid secretion seems to be complex.


Gender differences in tardive dyskinesia: a critical review of the literature.

We analyzed data from 76 selected studies on prevalence of tardive dyskinesia (TD), published through 1989. The primary focus was on gender differences. The overall prevalence of TD in the 39,187 patients included in these reports was 24.2 percent, and prevalence was significantly higher in women (26.6%) than in men (21.6%). The gender difference in TD prevalence appeared to narrow intriguingly in more recent studies. Overall, the TD prevalence seemed to reach its peak in the 50-70-year-old age group in men and continued to rise after age 70 in women. Also, women tended to have more severe TD than men. Spontaneous dyskinesia too was found to be more common in women. The material was also analyzed for cultural differences by comparing studies in four continents: North America, Europe, Africa, and Asia. Although grouping together studies from different countries in a continent into a single group is somewhat problematic, we found that Asian patients had lower prevalence of TD than North American, European, and African patients. Limitations of our review (including differences among studies in diagnostic criteria, observer bias, etc.) as well as possible explanations for the reported differences in the risk for TD are discussed.


Influence of acetylator status on sulphasalazine efficacy and toxicity in patients with rheumatoid arthritis.

The influence of acetylator status on the therapeutic efficacy and the toxicity of sulphasalazine (SASP) was assessed in 106 patients with rheumatoid arthritis (RA). Changes of indices of disease activity after 6 months, and progression of erosions after 2 years of SASP treatment were similar in fast and slow acetylators. Incidence and nature of withdrawals and side-effects, and requirement for intra-articular steroid injections or combination therapy due to poor response to SASP were almost identical in the two groups. A significant increase of the hepatic enzyme aspartate transaminase was noted mainly in slow acetylators, but was not associated with clinical disease. These results suggest that acetylator status does not relate significantly to either the efficacy or the toxicity of SASP in RA. It is possible that hepatic metabolism is affected by SASP, particularly in slow acetylators, but this does not lead to clinically identifiable problems.


Association of selected social, environmental and constitutional factors to blood lead levels in men aged 55-75 years.

Blood lead (B-Pb) levels were determined in 1802 out of 1856 non-occupationally exposed men aged 55-75 years living in the Rome area who participated, between 1989 and 1990, in an epidemiological survey for coronary heart disease (New Risk Factors Project). The median B-Pb level was 113 micrograms/l (10th-90th centiles: 74-180 micrograms/l) and only 0.7 per cent (n = 14) of the subjects had B-Pb values higher than 300 micrograms/l. B-Pb levels were significantly and positively associated to alcohol consumption. Moderate and heavy drinkers had median B-Pb level of 143 micrograms/l (10th-90th centiles: 92-233) and 165 micrograms/l (10th-90th centiles: 102-285) respectively, whereas non-drinkers had a median B-Pb level of 96 micrograms/l (10th-90th centiles: 66-143). The influence of smoking habits was less relevant. Subjects who never smoked and subjects smoking more than 20 cigarettes daily had median B-Pb levels of 103 and 133 micrograms/l, respectively. Individuals classified as habitual car-drivers had slightly higher Pb levels than non-drivers. Subjects classified as manual workers had higher B-Pb levels in comparison with non-manual workers and retired subjects. B-Pb levels were directly related to HDL-cholesterol (HDL-C, r = 0.2252) and gamma-glutamyltransferase (gamma-GT, r = 0.2207) serum levels. The alleged alcohol consumption was more related to B-Pb level (r = 0.3848) than to serum level of HDL-C (r = 0.2474) or gamma-GT (r = 0.2469). A significant correlation (r = 0.2409) also existed between B-Pb and blood cadmium levels (B-Cd). Subjects with a low Gaensler ratio, an index of respiratory function, had higher B-Pb levels. In multiple regression analyses alcohol intake was the most important predictor of B-Pb level, explaining more (14.27%) of the total variance than did B-Cd (4.98%), HDL-C (1.89%), driving habits (1.46%), gamma-GT (1.09%), skinfold thickness (0.96%), and Gaensler index (0.38%). The risk ratio of having B-Pb level higher than 180 micrograms/l (90th centile of B-Pb distribution in our subjects) was 5.3 (95% CI: 2.7-10.4) for drinkers versus non-drinkers and 1.9 (95% CI: 1.2-3.1) for current smokers versus subjects who had never smoked. B-Pb was, at least in our subjects, a more specific and sensitive objective index of alcohol consumption than gamma-GT and HDL-C.(ABSTRACT TRUNCATED AT 400 WORDS)


The human Y chromosome: overlapping DNA clones spanning the euchromatic region.

The human Y chromosome was physically mapped by assembling 196 recombinant DNA clones, each containing a segment of the chromosome, into a single overlapping array. This array included more than 98 percent of the euchromatic portion of the Y chromosome. First, a library of yeast artificial chromosome (YAC) clones was prepared from the genomic DNA of a human XYYYY male. The library was screened to identify clones containing 160 sequence-tagged sites and the map was then constructed from this information. In all, 207 Y-chromosomal DNA loci were assigned to 127 ordered intervals on the basis of their presence or absence in the YAC's, yielding ordered landmarks at an average spacing of 220 kilobases across the euchromatic region. The map reveals that Y-chromosomal genes are scattered among a patchwork of X-homologous, Y-specific repetitive, and single-copy DNA sequences. This map of overlapping clones and ordered, densely spaced markers should accelerate studies of the chromosome.


Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors.

Comparative genomic hybridization produces a map of DNA sequence copy number as a function of chromosomal location throughout the entire genome. Differentially labeled test DNA and normal reference DNA are hybridized simultaneously to normal chromosome spreads. The hybridization is detected with two different fluorochromes. Regions of gain or loss of DNA sequences, such as deletions, duplications, or amplifications, are seen as changes in the ratio of the intensities of the two fluorochromes along the target chromosomes. Analysis of tumor cell lines and primary bladder tumors identified 16 different regions of amplification, many in loci not previously known to be amplified.


Reductase activity encoded by the HM1 disease resistance gene in maize.

The HM1 gene in maize controls both race-specific resistance to the fungus Cochliobolus carbonum race 1 and expression of the NADPH (reduced form of nicotinamide adenine dinucleotide phosphate)-dependent HC toxin reductase (HCTR), which inactivates HC toxin, a cyclic tetrapeptide produced by the fungus to permit infection. Several HM1 alleles were generated and cloned by transposon-induced mutagenesis. The sequence of wild-type HM1 shares homology with dihydroflavonol-4-reductase genes from maize, petunia, and snap-dragon. Sequence homology is greatest in the beta alpha beta-dinucleotide binding fold that is conserved among NADPH- and NADH (reduced form of nicotinamide adenine dinucleotide)-dependent reductases and dehydrogenases. This indicates that HM1 encodes HCTR.


Nucleotide-iron-sulfur cluster signal transduction in the nitrogenase iron-protein: the role of Asp125.

Electron transfer in nitrogenase involves a gating process initiated by MgATP (magnesium adenosine triphosphate) binding to Fe-protein. The redox site, an 4Fe:4S cluster, is structurally separated from the MgATP binding site. For MgATP hydrolysis to be coupled to electron transfer, a signal transduction mechanism is proposed that is similar to that in guanosine triphosphatase proteins. Based on the three-dimensional structure of Fe-protein, Asp125 is likely to be part of a putative transduction path. Altered Fe-protein with Glu replacing Asp has been prepared and retains the ability for the initial nucleotide-dependent conformational change. However, either MgADP or MgATP can induce the shift and Mg binding to the nucleotide is no longer essential.


Prevention of protein denaturation under heat stress by the chaperonin Hsp60.

The increased synthesis of heat shock proteins is a ubiquitous physiological response of cells to environmental stress. How these proteins function in protecting cellular structures is not yet understood. The mitochondrial heat shock protein 60 (Hsp60) has now been shown to form complexes with a variety of polypeptides in organelles exposed to heat stress. The Hsp60 was required to prevent the thermal inactivation in vivo of native dihydrofolate reductase (DHFR) imported into mitochondria. In vitro, Hsp60 bound to DHFR in the course of thermal denaturation, preventing its aggregation, and mediated its adenosine triphosphate-dependent refolding at increased temperatures. These results suggest a general mechanism by which heat shock proteins of the Hsp60 family stabilize preexisting proteins under stress conditions.


Dual-target inhibition of HIV-1 in vitro by means of an adeno-associated virus antisense vector.

An adeno-associated virus vector encoding an antisense RNA was used to transduce stable intracellular resistance to human immunodeficiency virus-1 (HIV-1) in human hemopoietic and non-hemopoietic cell lines. The antisense targets are present in all HIV-1 transcripts and include the TAR sequence, which is critical for transcription and virus replication, and the polyadenylation signal. Cell lines expressing antisense RNA showed up to 95 percent inhibition of gene expression directed by the HIV-1 long terminal repeat and greater than 99 percent reduction in infectious HIV-1 production, with no detectable cellular toxicity. Because of their efficient transcription and inability to recombine with HIV-1, adeno-associated virus vectors represent a promising form of anti-retroviral gene therapy.


The time course of glutamate in the synaptic cleft.

The peak concentration and rate of clearance of neurotransmitter from the synaptic cleft are important determinants of synaptic function, yet the neurotransmitter concentration time course is unknown at synapses in the brain. The time course of free glutamate in the cleft was estimated by kinetic analysis of the displacement of a rapidly dissociating competitive antagonist from N-methyl-D-aspartate (NMDA) receptors during synaptic transmission. Glutamate peaked at 1.1 millimolar and decayed with a time constant of 1.2 milliseconds at cultured hippocampal synapses. This time course implies that transmitter saturates postsynaptic NMDA receptors. However, glutamate dissociates much more rapidly from alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. Thus, the time course of free glutamate predicts that dissociation contributes to the decay of the AMPA receptor-mediated postsynaptic current.


Elephantiasis: a disease of development in north east Ghana.

A reconnaissance survey for the presence of lymphatic filariasis is made in 41 chiefdoms of north east Ghana. Four disease levels are identified culminating in hyperendemic disease foci associated with two Government-introduced rice irrigation projects. Attention is also drawn to the disease effects of small village dams. Multiple concurrent infections are noted. Within the most stricken irrigation villages, aspects of concealment, stigma and marriage are considered. Failure to control lymphatic filariasis has led to hospital avoidance and neglect of the disease jointly by patients, physicians and nurses. Culpability rests with the irrigation authority and Government health services. An outline is given of possible measures for disease control. A multisectoral policy of 'prevention before development' is strongly advocated.


Assessment of sanitation conditions by qualitative sanitation measurement.

Survey of sanitation conditions by qualitative sanitation measurement were carried out in three districts, Chachoengsao Province, Thailand. The sanitation of each household was assessed in terms of positive stool examination and scores of seven main sanitary activities which were used to classify each household's sanitation as hygienic or non-hygienic. Stool collections were made from each household's housewife and examined for pathogenic bacteria and parasites. It was found that in the qualitative measurement of the household's sanitation as hygienic or non-hygienic, most sanitary activities were highly associated with the results of stool examination. From consideration of the sensitivity, specificity and kappa coefficient of significant sanitary activities as predictors and the result of stool examination was used as gold standard, it was apparent that a package of all main sanitary activities was the most appropriate measurement in the survey to assess sanitation conditions in the community.


Beta-cyfluthrin, a synthetic pyrethroid for mosquito control.

Beta-cyfluthrin (OMS 3051), a new synthetic pyrethroid and one of the stereoisomers of cyfluthrin, was studied for insecticidal activity against eight mosquito species. Its larvicidal activity with LC50 values of 5.62 x 10(-5) and 1.19 x 10(-4) mg/l respectively for Culex quinquefasciatus and Aedes aegypti was comparable with that of deltamethrin. This pyrethroid was more effective against the larvae of Armigeres subalbatus (LC50 - 7.76 x 10(-7)) and the adults of Anopheles culicifacies LT50 - 27.76 min at 2.0 mu/cm2) than the other species tested. Residual efficacy at 50 mg(ai)/m2 was more persistent (for 14-25 weeks) on thatch and asbestos among the four treated surfaces. This compound also elicited oviposition deterrent activity at 0.001 mg/l against Cx.quinquefasciatus. beta-cyfluthrin is a good insecticide for mosquito control. However, care should be exercised while using it as a larvicide in breeding habitats considering its toxicity to fish.


The resting and house frequenting behavior of Mansonia annulifera, Ma. uniformis and Ma. indiana, the vectors of Malayan filariasis in Kerala State, India.

Mansonia annulifera, was recorded to be an endophilic species, preferring to rest indoors, while Ma. uniformis was exophilic, having a predilection for outdoor resting habitats, eg bushes and shrubs. Ma. indiana did not show a clear preference to either of these biotopes. In indoor resting collections, the unfed proportion of Ma. uniformis was significantly higher during post-dusk compared to day hours (p < 0.05), indicating that this exophilic species enters houses during dusk hours for feeding. The full fed proportion was higher during day hours compared to dusk/night hours. The semigravid proportion showed a significant reduction during post-dusk hours (p < 0.05). These findings suggest that after having a blood-meal this species rest indoors and leave the houses for outdoor resting sites during the dusk hours on the subsequent night.


Evaluation of carbon dioxide and 1-octen-3-ol as mosquito attractants.

CDC Light traps were used to study the attractant effect of CO2 and 1-octen-3-ol on trap catches of mosquito populations at three different locations in Malaysia. There was a significant increase in the number of mosquitos caught in traps baited with CO2 and CO2 with 1-octen-3-ol. The number of mosquitos caught in the CDC light trap and in the CDC light trap baited with 1-octen-3-ol alone were very few. 1-octen-3-ol and CO2 acted synergistically in attracting significantly greater numbers of Culex tritaeniorhynchus. However Anopheles sp. were not very attracted to light traps even with attractants added to them.


Factors affecting compliance with depot injection treatment in the community.

This study investigated aspects of the community psychiatric care of psychotic patients living in inner city areas, and especially those of Afro-Caribbean ethnicity. Ethnicity was not found to be a major factor affecting compliance with depot injections. More important were variables such as time since first treatment and sex. Defaulters from treatment were more likely to be subsequently admitted to hospital.


Complete hamster CAD protein and the carbamylphosphate synthetase domain of CAD complement mammalian cell mutants defective in de novo pyrimidine biosynthesis.

The mammalian CAD gene codes for a 240-kDa multifunctional protein that catalyzes the first three steps of de novo pyrimidine biosynthesis. Previously, the longest cDNA construct available was missing approximately 500 bp of coding sequence at the 5' end, thereby lacking the sequence to encode the entire carbamylphosphate synthetase (CPSase) domain. Here, a complete CAD hamster cDNA is constructed, placed into a mammalian expression vector, and transfected into hamster cells deficient in CAD. Transfectants show coordinately restored levels of all three enzyme activities and the presence of full-length CAD protein. A derivative construct of the CAD cDNA was generated that should encode only the CPSase domain. When transfected into mammalian cells, a protein was synthesized that had significant CPSase activity both in vivo and in vitro. The two constructs generated in this study will facilitate the study of CAD structure, function, and allosteric regulation.


Spinal epidural abscess: a report of 40 cases and review.

Despite modern medical advances, the morbidity and mortality rates associated with spinal epidural abscess remain significant, and the diagnosis often is elusive. A retrospective study was undertaken to define better the incidence and clinical features of this infection, and to establish current diagnostic and therapeutic guidelines. Forty cases of spinal epidural abscess were encountered at our institution between July 1979 and March 1991. All medical records and radiological images were reviewed. We report a significant increase in the incidence of epidural abscess after June 1988 (p = 0.0195). Sixteen patients used drugs intravenously, and six had undergone spinal procedures. Twelve patients were misdiagnosed in various emergency rooms or clinics and discharged. Localized back pain, fever, and neurological deficit remained the typical clinical manifestations. Erythrocyte sedimentation rate was elevated uniformly when measured (21 cases). Magnetic resonance imaging was diagnostic specifically in 23 of 24 instances. The majority of patients underwent surgical drainage, but five selected patients were managed nonoperatively. The highly variable presentation of spinal epidural abscess may confuse the diagnosis and delay indicated surgical intervention. Localized back pain in a febrile patient at significant risk for epidural abscess warrants erythrocyte sedimentation rate measurement. The presence of erythrocyte sedimentation rate elevation or evidence of spinal cord compression on physical examination are indications for immediate magnetic resonance imaging examination with contrast enhancement. Surgical drainage with sustained intravenous antibiotic treatment remains the cornerstone of therapy. Nonoperative management may be considered in selected cases.


[The neuroendocrine reactions in patients with chronic bronchitis in a region of the northeastern USSR].

The author describes differences in the manifestations of mutually determined reactions of the sympathoadrenal and hypophyseoadrenocortical systems in their interaction with the biologically active substance serotonin in healthy persons (124) and patients suffering from chronic bronchitis at different times of living on the Chukot Peninsula. The most perfect system of the protective effect on the part of the neuroendocrine complex can be seen in chronic bronchitis patients within the first 3-5 years of living in the North as compared with the reaction of the neuroendocrine system in the examined patients with a longer "northern record" (10-15 years and more).


Transplacental exposure to tobacco smoke in human-adduct formation in placenta and umbilical cord blood vessels.

Smokers are exposed to a large number of genotoxic compounds that react with DNA to form covalently bound carcinogen-DNA adducts after metabolic conversion to their biological active form. Using the P32-postlabeling techniques, tobacco smoke related carcinogen--DNA adducts have been demonstrated in DNA isolated from human placenta and umbilical cord vein and artery obtained from 11 nonsmoking and 8 smoking normal healthy women and foetuses. The adduct level was significantly higher in tissues from smokers than from nonsmokers (P = 0.021), when all tissues were combined. Furthermore, the total adduct level was higher in maternal tissue than the level in fetal tissues (P = 0.030). The adduct level in umbilical cord vein DNA was significantly lower than in placenta, and marginally lower than in umbilical cord artery from the same donor. This suggests that the foetus can metabolise some of the genotoxic compounds found in tobacco smoke to DNA-binding metabolites. The presence of DNA adducts in foetal tissues is indicative of potential genomic damage, that may result in an increased risk for the development of serious diseases, like cancer in childhood or later during the life span of the individual.


Effects of reproductive tract glutathione enhancement and depletion on ethyl methanesulfonate-induced dominant lethal mutations in Sprague-Dawley rats.

The effects of altering glutathione (GSH) levels in the male reproductive tract have been studied in an attempt to establish a link between chemical-induced perturbations in glutathione and susceptibility of spermatozoa to chemical insult. Tissue GSH levels were enhanced by a treatment regimen of N-acetylcysteine (NAC) (250 mg/kg, 4 treatments at 2 h intervals). With this treatment, GSH levels in liver, testis, caput epididymis, and cauda epididymis were elevated to 126%, 110%, 178%, and 136% of control values. Sexually mature male rats were then treated with NAC and challenged with a dose of EMS (100 mg/kg) to determine if enhanced tissue GSH would protect against EMS-induced dominant lethal mutations. Pretreatment with NAC significantly decreased the post-implantation loss from 7.05 +/- 0.57 with EMS alone to 5.28 +/- 0.47. Conversely, a dominant lethal assay was conducted using different doses of phorone pretreatment to determine the relative contribution of hepatic versus reproductive tract GSH in protecting against EMS-induced dominant lethal resorptions. Doses of 100 mg/kg and 250 mg/kg phorone significantly lowered both hepatic and reproductive tract GSH while 25 mg/kg lowered only hepatic GSH. These three dose levels were used as pretreatments in a dominant lethal study followed by a challenge administration of EMS (50 mg/kg), which is a threshold dose of EMS for producing dominant lethal mutations. Comparison against controls demonstrated a significant potentiation of fetal resorptions in all groups receiving phorone pretreatment, including the 25 mg/kg pretreatment group which only lowered hepatic GSH prior to EMS challenge. The results of these experiments indicate that GSH reserves in the male reproductive tract are insufficient to protect developing spermatozoa from damage by alkylating agents in the absence of hepatic GSH.


Lack of synergism among DMAB, BOP, and MNU in induction of carcinomas of the rat ventral prostate.

The present experiment was undertaken to investigate possible synergism in induction of rat prostate tumor. F344 rats were repeatedly administered the prostate carcinogens 3,2'-dimethyl-4-aminobiphenyl (DMAB),N-nitrosobis(2-oxopropyl)amine(BOP), and N-methylnitrosourea (MNU) sequentially or together at times of hormonal castration induced regenerative cell proliferation of prostate epithelial cells. Group 1 animals received two 100 mg/kg doses of DMAB, two 20 mg/kg applications of BOP, and two times 15 mg/kg of MNU. Groups 2, 3, and 4, respectively, received each of the carcinogen treatment alone. Group 5 was given 6 applications of all three in combination, each at one-third the dose administered to the other groups. The results showed a clear synergism in enhanced tumor development in the colon but not in the prostate, in which the incidence of lesions induced by the three carcinogens was similar to that with DMAB alone.


Strategy of research for cancer--chemoprevention.

It has been reported that environmental chemicals are important factors in terms of both development and prevention of human cancer. For the latter, detection of early stages is an essential first step followed by clinical trials for surveying populations at risk. Thus a great deal of attention has been focused on these areas. However, investigations of possibilities for active prevention of cancer development itself form another major project. Chemoprevention of carcinogenesis, which means prevention of carcinogenesis by exogenous chemical compounds, has been investigated extensively in a variety of organs in animal models. Usually attention is concentrated on only one organ. However, antioxidants, such as BHA, exert very different effects on different organs, suggesting the necessity of whole body approaches to the question of chemoprevention. Furthermore, the mechanisms of chemoprevention, including the step of carcinogenesis, i.e., initiation, promotion, progression or whole carcinogenesis steps, in which exogenous compounds exert their protective effects, should be considered. A medium-term bioassay system and a multi-organ carcinogenesis system, which can be used for investigation of potential for cancer chemoprevention, have been developed in our laboratory. Dose dependent inhibitory effects were established for both BHA and alpha-tocopherol in the medium-term bioassay system, and inhibition of small intestinal carcinogenesis by catechins in green tea has also been investigated in our multi-organ carcinogenesis protocol. It is extremely important for prevention of human cancer that we find new candidates for chemopreventive agents using animal studies. This paper reviews published reports on chemoprevention, taking into account effective stages, and proposes suitable experimental animal models for future investigations in this increasingly important area.


Airway calibre as a confounder in interpreting bronchial responsiveness in asthma.

The relation between airway responsiveness to constrictor agents and forced expiratory volume in one second (FEV1) is important when interpreting change in airway responsiveness after an intervention. The aim of the study was to analyse the relation between FEV1 as a percentage of predicted values (% predicted) and airway responsiveness between and within asthmatic subjects. Results of non-specific bronchial challenge tests were pooled from two randomised crossover studies comparing the effect of a non-sedative antihistamine with placebo in 35 patients with moderate asthma. The design of the two studies was similar: the provocative concentration of either histamine (first study) or methacholine (second study) resulting in a 20% decrease in ventilatory capacity (PC20) was repeated at two week intervals while patients were treated with the antihistamine or placebo. The dose of inhaled corticosteroid was gradually reduced during the study. Data were analysed with PC20 as the dependent variable in a general linear model so that the influence on PC20 of inhaled corticosteroid dose, antihistamine, and choice of bronchoconstricting agent could be separated from the influence of FEV1% predicted. The correlation coefficient between mean PC20 and mean prechallenge FEV1 for each patient was 0.45. In the general linear model two thirds (65%) of the variation in PC20 was due to variation between subjects. One third of the within subject variation in PC20 could be explained by variation in prechallenge FEV1% predicted (a change in FEV1 of 27% predicted was associated with one doubling or halving of PC20). Treatment with the antihistamine had no influence on PC20, except when histamine was used as the bronchoconstricting agent. The dose of inhaled corticosteroid had a small but significant effect. The variation in a patient's PC20 over time (several months) is related to changes in FEV1% predicted. Variation in FEV1% predicted explains less of the variation in bronchial responsiveness between subjects where a patient specific factor, which is probably related to the pathogenesis of bronchial asthma, seems to dominate.


Development and evaluation of ELISAs for factor XIIIA and XIIIB subunits in plasma.

Severe, congenital deficiency of factor XIII is extremely rare. However, a moderate reduction in the plasma level of the functional subunit (factor XIIIA) and also to a lesser extent of the carrier subunit (factor XIIIB), and a decrease in the XIIIA:B subunit ratio, have recently been reported in patients with the inflammatory bowel disorder Crohn's disease, particularly during clinical relapse. In order to accurately monitor patients, sensitive, reliable assays for the two subunits of factor XIII are required. We report here the development and validation of ELISAs for these components. The assays are identical except in respect of the specificity of the polyclonal antiserum used as starting material, both of which are commercially available. The antisera are purified by n-octanoic acid precipitation and portions of these purified immunoglobulins are used as coating antibodies. The remaining portions are biotinylated and used with streptavidin and horse-radish peroxidase as tracer antibodies. A normal range (n = 24) was established for factor XIIIA (mean 95 range 60-130 U/dl) and for factor XIIIB (mean 99 range 60-130 U/dl). There were no significant differences between the ELISA and electroimmunodiffusion assays either for factor XIIIA (means +/- 1 standard deviation 95 +/- 15.9 and 89 +/- 22.7 respectively) or for factor XIIIB (99 +/- 18.3 and 106 +/- 23.4 respectively). These assays have been in routine use for six months, during which time two further antisera purifications and biotinylations have been carried out without significant problems of reproducibility or stability.


Canine leukocyte cell adhesion molecules (LeuCAMs): characterization of the CD11/CD18 family.

The canine LeuCAM (CD11/CD18) family is characterized using a panel of newly developed anti-canine LeuCAM monoclonal antibodies (mAb) as well as a cross-reactive anti-human CD11b mAb. The new anti-canine LeuCAM mAb were developed by immunizing mice with purified canine CD11/CD18 antigen derived from an anti-canine CD18 affinity column. Six mAb with reactivity to the canine LeuCAM family were cloned and characterized. These 6 included a new anti-canine CD18 mAb, 2 anti-canine CD11a mAb, 2 anti-canine CD11c mAb, and a mAb which recognizes a novel canine CD11/CD18-related macrophage antigen, designated LeuCAMmac, which is described in a separate report. The cell and tissue distribution of canine LeuCAMs was found to be similar to that reported in other species with a few notable exceptions. One prominent exception was the finding that canine CD11c, in contrast to human CD11c, was much more widely distributed on dendritic cells than it was on tissue macrophages. The molecular organization of the canine LeuCAM family was also found to be similar to that reported in other species, with the possible exception of the canine CD11b alpha chain, which may exist as several different species. It is anticipated that these anti-canine LeuCAM mAb will prove useful in immunophenotypic studies of canine hemolymphatic system neoplasia.


Association of HLA-B51 and lack of association of class II alleles with Behçet's disease.

Eighty-one Behcet's disease patients have been studied for HLA association by HLA-DRB1, -DQA1, -DQB1 and -DPB1 genotyping with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique and for NcoI RFLP in the tumor-necrosis factor (TNF beta) gene by Southern hybridization in addition to serological typing. In serological typing, the frequency of HLA-B51 was significantly increased in the patients. In PCR genotyping, there was a significant increase in the HLA-DRB1*0802, DQA1*0301 and DQB1*0303 alleles, whereas the frequencies of DRB1*1502, DQA1*0103, DQA1*0101, DQB1*0601 and DQB1*0501 showed a significant decrease in the patients. No significant difference was observed in any HLA-DPB1 alleles. Among them, B51 was found to be a genetic marker most strongly associated with Behcet's disease (p less than 0.00005, chi 2 = 46.47, pc[corrected p] less than 0.005). The positive and negative associations of class II alleles with the disease can be explained by linkage disequilibrium with B51, and do not reach statistical significance by the corrected p-value test. In NcoI RFLP typing in the TNF beta gene, 250 kb centromeric of the HLA-B gene, the frequency of a 5.5 kb fragment was considerably decreased in the patients when compared to the controls, although the decrease was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)


HLA-DQB1 allele typing by a new PCR-RFLP method: correlation with a PCR-SSO method.

We have developed a new PCR-RFLP method for HLA-DQB1 typing. This method was easy to follow, requiring only one DQB1 generic amplification and 5 endonucleases to assign 14 out of 15 HLA-DQB1 alleles. In addition, we determined that by using one generic amplification and two enzymes (Sau96 I and Hae III) it was possible to type the generic specificities: DQw2, DQw4, DQw5, DQw6, and DQw7, DQw8-9, providing a practical alternative for serological HLA-DQ generic typing. We also performed a side-by-side correlation with a PCR-SSO typing method and found an almost 100% concordance between the methods. The limitations of these methods were: 1) the PCR-RFLP method did not allow the differentiation between the HLA-DQB1*0602 and *0603 alleles; 2) the PCR-SSO method gave crosshybridization signals in the detection of *0302 or *0303 alleles. Our results suggested that both methods, PCR-RFLP and PCR-SSO, are useful alternatives for HLA-DQB1 typing.


Studies of RFLP-inferred HLA-DR-DQ haplotypes in Danish women with recurrent fetal losses.

Results of HLA-DR and -DQ typing by RFLP in 152 unrelated women with at least 3 unexplained fetal losses were compared with those of 210 normal controls. The overall distribution of DR-DQ phenotypes did not differ significantly between patients and controls. In a subgroup of 59 patients having had at least 4 fetal losses, a significantly higher number of patients than controls were DRw17,DQw2-positive (RR = 2.9, pcorrected less than 0.02). DR-DQ haplotypes which carry DQB1 alleles encoding an amino acid other than aspartic acid at codon 57 in the second exon (non-Asp57 haplotypes) have been reported to confer susceptibility to several immunologically-mediated diseases. We found that the total frequency of non-Asp57 haplotypes (other than DR7,DQw2 which has unique features) was significantly increased in patients (50.0%) compared with controls (39.3%) (p less than 0.005). In the total group of patients and in the group with at least 4 fetal losses, 22.4% and 32.2% respectively were homozygous non-Asp57/non-Asp57 compared with 13.3% among controls (RR = 1.9, p less than 0.025 and RR = 3.1, p less than 0.001, respectively). The results suggest that DRw17,DQw2 confers susceptibility to suffer recurrent fetal losses. However, it is possible that the HLA-associated susceptibility may be primarily conferred by DQ molecules lacking aspartic acid at codon 57 in the DQ beta chain.


HLA-DPB1 typing with polymerase chain reaction and restriction fragment length polymorphism technique in Danes.

We have used the polymerase chain reaction (PCR) in combination with the restriction fragment length polymorphism (RFLP) technique for HLA-DBP1 typing. After PCR amplification of the polymorphic second exon of the HLA-DPB1 locus, the PCR product was digested with seven allele-specific restriction endonucleases: RsaI, FokI, ApaI, SacI, BstUI, EcoNI, and DdeI, and the DNA fragments were separated by electrophoresis in agarose gels. Altogether, 71 individuals were investigated and 16 different HLA-DPB1 types were observed in 26 different heterozygotic combinations, as well as five possible homozygotes. Four heterozygotes could not be unequivocally typed with the PCR-RFLP method. The HLA-DPB1 typing results obtained with the PCR-RFLP method were compared with the typing results obtained with PCR allele-specific oligonucleotides (PCR-ASO) in 50 individuals. The results obtained with the two methods were concordant in 84% of the cases. One of the HLA-DPB1 types was discrepant in six heterozygotes, both HLA-DPB1 types were discrepant in one heterozygote, and in one individual two HLA-DPB1 types were identified with the PCR-RFLP technique while only one HLA-DPB1 type could be demonstrated with the PCR-ASO technique. The frequencies of the HLA-DPB1 genotypes deduced from the results of PCR-RFLP typing were estimated in 71 healthy Danes.


A mosquito neuropeptide in a moth larva (Helicoverpa zea): relation to FMRF-amide immunoreactivity.

The cerebral nervous and midgut endocrine systems of the larval corn earworm, Helicoverpa zea, were examined using light microscopy and immunocytochemistry for RF-amide family peptides. Immunoreactivity for a mosquito neuropeptide, Aedes Head Peptide-I (Aea-HP-I,pERPhPSLKTRFa), is widely distributed in this lepidopteran. Immunostaining for Aea-HP-I is localized (1a) in perikarya and axons of the brain, the subesophageal ganglion, and the first thoracic ganglion, (b) in peripheral axons innervating muscles of the midgut, and (2) in numerous midgut endocrine cells. Aea-HP-I-associated activity generally occurs as a subset of FMRF-amide (FMRFa; a molluscan cardioactive peptide) immunoreactivity. Cross-reactivity studies indicate that Aea-HP-I and FMRFa immunoreactivities are heterogeneous in the cerebral nervous system and in axons innervating the muscles of the midgut, but may be homogeneous in midgut endocrine cells. Radioimmunoassay for Aea-HP-I reveals immunoreactivity in hemolymph, as well as in extracts of midguts and heads.


Morphological alterations accompanying the effect of peptaibiotics, alpha-aminoisobutyric acid-rich secondary metabolites of filamentous fungi, on Culex pipiens larvae.

The effect of different representatives of the group of peptaibiotics, alpha-amino-isobutyric acid rich secondary metabolites of filamentous fungi, on Culex pipiens larvae was studied. Light and transmission electron microscopy techniques were used to localize the intracellular damage and to determine the target organells for the mode of action of peptaibols in mosquito larvae. Though different in insecticidal activity, all tested compounds induced the same type of tissue damage, which was characterized by heavy challenge of mitochondria followed by partial swelling, crystaeolysis and destruction of mitochondrial walls. It is concluded that the mode of action of peptaibols in mosquito larvae is mediated through the damage of mitochondria. The structure-mosquitocidal effect of these compounds, their potential mode of action and role in the natural fungal entomopathogenic process are briefly discussed.


[Old and new alpha 2-adrenoceptor agonists. 1. Xylazine and medetomidine].

The "discovery" of alpha 2-receptors permits the explanation of the mechanism of xylazine that was hitherto poorly understood. The substance can be classified as an alpha 2-adrenoceptor agonist. In the near future two new alpha 2-adrenoceptor agonists will be available in Germany: medetomidine for small animals and detomidine for large animals. Also a new alpha 2-adrenoceptor antagonist will be introduced: atipamezole. alpha 2-adrenoceptor agonists cause sedation, analgesia and muscle relaxation. Notable side effects are bradycardia, heart block and a dose dependent respiratory depression. A comprehensive survey on the pharmacology of the alpha 2-adrenoceptor agonists is given. The previous literature is summarized and discussed, including own studies and clinical experience. An attempt is made to assess the clinical importance of the new drugs. For certain indications medetomidine alone may not be satisfying and combination with other drugs becomes necessary. If sedation is inadequate, the combination with benzodiazepines or propofol is advised; if analgesia is poor, the combination with ketamine or an opioid is recommended.


Sertoli cells isolated from adult 2,5-hexanedione-exposed rats exhibit atypical morphology and actin distribution.

Sertoli cells were isolated from 2,5-hexanedione (2,5-HD)-exposed, cryptorchid and 21-day-old rats in order to examine alterations in in vitro Sertoli cell transferrin secretion, germ cell adhesion, in vitro morphology, and cytoskeletal organization which might be involved in the irreversibility of 2,5-HD-induced testicular injury. Sertoli cells isolated from 21-day-old, cryptorchid and 2,5-HD-exposed rats exhibited similar transferrin secretion as measured using an enzyme-linked immunosorbent assay. Germ-cell adhesion was measured using [3H]leucine-labeled immature rat germ cells and revealed similar levels of germ-cell binding in Sertoli cell cultures isolated from the three groups of rats. Differential interference contrast microscopy demonstrated that Sertoli cells isolated from 2,5-HD-exposed rats possessed an atypical spindle shape and long cytoplasmic processes. The immunofluorescent distribution of tubulin and vimentin corresponded with the morphological appearance of the cells with well-defined microtubule and intermediate filament networks which, in the cells isolated from 2,5-HD-exposed rats, extended into the cytoplasmic processes. Rhodamine-conjugated phalloidin-labeled actin stress fibers were decreased in density within the 2,5-HD-exposed rat Sertoli cells. The altered morphology and distribution of actin filaments within Sertoli cells isolated from adult 2,5-HD-exposed rats may reflect an underlying insult which is involved in the irreversible nature of 2,5-HD intoxication.


The effects of Indian red scorpion Buthus tamulus venom in vivo and in vitro.

The Indian red scorpion Buthus tamulus (or Mesobuthus tamulus) can cause fatal envenoming, but its mechanism of action is unclear. Venom was tested in vivo in anaesthetized rats and in vitro on isolated cardiac and skeletal muscle preparations. In vivo, the venom caused marked rhythmical fluctuations in blood pressure preceding cardiovascular collapse and death. On sheep Purkinje fibres, venom could induce spontaneous action potentials and cause prolongation of action potential duration. In chick biventer cervicis and mouse triangularis sterni preparations, venom enhanced the release of acetylcholine and induced repetitive firing of nerve action potentials in response to single shock stimulation. High concentrations caused stimulation then block of neuromuscular transmission. The main effects of Buthus tamulus venom are likely to be due to toxins that affect the opening of Na+ channels in nerves and muscles. This will cause an increase in the release of neurotransmitters in the peripheral nervous system, which may produce cardiovascular abnormalities and respiratory paralysis.


Purification and characterization of iotrochotin, a novel toxin from the Caribbean sponge Iotrochota birotulata, which selectively permeabilizes synaptosomes.

A protein termed iotrochotin (IOT) was isolated from the exudate of the Caribbean sponge Iotrochota birotulata using as an assay its stimulation of the release of radioactivity from synaptosomes preloaded with [3H]choline. Sephadex G-50 chromatography of the exudate produced one peak, with a mol. wt of approximately 18,000, which was further resolved into two active fractions by anion exchange chromatography. The more tightly bound of the two fractions was characterized and referred to as IOT. The action of IOT was essentially complete by 0.5-1.0 min and was independent of the Ca2+ or Na+ content of the incubation mixture. Released radioactivity included about 50% each of [3H]acetylcholine and [3H]choline. Release of radioactivity increased as a function of IOT concentration and then reached a maximum. Extrapolated asymptotic release was nearly equal to that obtained by lysing the synaptosomes. IOT also released radioactivity from synaptosomes which had been preincubated with other tritiated neurotransmitters or with 2-[3H]deoxy-D-glucose. Lactate dehydrogenase and choline acetyltransferase activities were not released from synaptosomes by treatment with IOT, but were released by digitonin. IOT therefore releases some of the smaller molecular weight components of synaptosomes, but does not permeabilize the synaptosomal membrane in the same way as digitonin treatment.


Phoneutria nigriventer (armed spider) venom induces increased vascular permeability in rat and rabbit skin in vivo.

The effect of intradermally injected Phoneutria nigriventer venom (PNV) on vascular permeability of both rat and rabbit skin has been investigated. Oedema formation was measured as the local extravascular accumulation at skin sites of intravenously injected 125I-human serum albumin. In both rat and rabbit PNV induced dose-dependent oedema which was greatly potentiated by the vasodilators calcitonin-gene-related peptide and prostaglandin E1. In rats, PNV-induced oedema was markedly reduced either by previous treatment of the animals with the histamine H1 antagonist mepyramine and the serotonin antagonist methysergide or when venom was dialysed, indicating a major role for histamine and serotonin. In rabbits, dialysis of the venom to remove histamine and serotonin did not reduce PNV-induced oedema, indicating presence of oedematogenic component(s) which are different from the amines histamine or serotonin.


Role of H2-receptor antagonists in the treatment of duodenitis.

Non-erosive duodenitis is an ulcer-independent disorder, the pathogenesis of which is still unclear but apparently unrelated to gastric hyperacidity. However, antisecretory agents such as H2-blockers can be effective not only in relieving dyspeptic symptoms but also in promoting endoscopic healing or improvement. In this respect conflicting data are reported with cimetidine while promising, although preliminary, results were obtained with ranitidine and with nizatidine. Nizatidine seems especially effective when administered at a dose of 150 mg b.i.d., a regimen providing moderate, but continuous acid inhibition throughout a 24 hour period.


The effect of donor T lymphocytes and total-body irradiation on hemopoietic engraftment and pulmonary toxicity following experimental allogeneic bone marrow transplantation.

To study the effects of donor T lymphocytes on engraftment and graft-versus-host disease in relation to recipient total-body irradiation, we have returned small numbers of T cells to T-cell-depleted bone marrow transplanted across a minor histocompatibility barrier in mice (B10.BR-->CBA). T-cell-depleted B10.BR marrow (10(7) cells) was transplanted into CBA recipients prepared with TBI doses ranging from 4 to 14 Gy. Selected animals also received 10(4) (0.1%) and 10(5) (1.0%) measured B10.BR T lymphocytes. The extent of donor marrow engraftment was determined from hemoglobin and carbonic anhydrase phenotyping of peripheral blood at 3 months posttransplant. Toxicity was assessed from breathing-rate measurements, histopathology, and animal survival. Addition of T cells had a profound effect on survival related to radiation dose. The TBI doses resulting in an LD50 at 12 weeks were 6.9 Gy, 9.3 Gy, and 13.0 Gy for animals receiving 10(5), 10(4), and no T cells, respectively. Mortality was associated with pulmonary dysfunction as measured by an elevation of breathing rates. Autopsy and histological analysis revealed extensive damage to the lung parenchyma. In contrast to the toxicity data, addition of T cells to the donor marrow had no effect on the TBI dose required for equivalent erythroid engraftment. These results demonstrate that in combination with TBI small numbers of T cells in the transplanted marrow do not aid engraftment but do significantly increase the risk of pulmonary toxicity.


Induction of a cell-transferable suppression of alloreactivity by photodamaged lymphocytes.

We have reported previously that splenocytes from BALB/c mice acutely rejecting CBA/j skin allografts were exposed to 8-methoxypsoralen (8-MOP) and ultraviolet A light and infused several times intravenously into naive BALB/c recipients; the recipients were hyporesponsive to CBA/j alloantigens in skin graft and delayed-type hypersensitivity assays, as well as in mixed leukocyte culture and cytotoxicity assays. We currently expand on this work by showing that donor-specific tolerance can be transferred adoptively to naive syngeneic animals via unfractionated splenocytes from mice rendered tolerant by the previous protocol. This suppressed response to alloantigen was transferred optimally with splenocytes taken from mice on the sixth day after the final treatment with PET cells. We further demonstrate that the cells that are adoptively transferring suppression are radiosensitive, Thy-1+, Lyt-2+, L3T4- T lymphocytes.


P-glycoprotein but not topoisomerase II and glutathione-S-transferase-pi accounts for enhanced intracellular drug-resistance in LoVo MDR human cell lines.

The biochemical bases of the multidrug-resistant (MDR) phenotype were investigated in drug-resistant sublines derived from LoVo human colon carcinoma cell lines by doxorubicin (DOX) and teniposide (VM26) selection. In addition to P-glycoprotein-mediated drug extrusion through the plasma-membrane, LoVo MDR cells display a further drug-resistance mechanism. That is, to achieve equitoxic effects, LoVo MDR sublines require much higher intracellular drug concentrations than those required by LoVo drug-sensitive parent cell line. Involvement of mdr1, topoisomerase II and glutathione-S-transferase-pi (GST-pi) drug-resistance systems in intracellular drug resistance was investigated. Pharmacologic and biochemical data indicated that intracellular drug resistance in LoVo MDR sublines is uniquely consequent to the drug-transporting property of intracytoplasmic membrane-bound P-glycoprotein molecules which compartment drugs in vacuole-like structures.


[Poisonings by beta-adrenoblockaders].

The review includes data on symptomatology of acute poisoning with beta-adrenoblockers, their distribution in biological material, main pathways of metabolism in human body. Necessity for developing methods of systemic analysis of beta-adrenoblockers in biological material is pointed out.


The reservoir of Plasmodium falciparum malaria in a holoendemic area of western Kenya.

The reservoir of infectious Plasmodium falciparum gametocytes in a population living in an area of holoendemic malaria in western Kenya was estimated by directly feeding mosquitoes on volunteers. Resulting mosquito infections were assessed both by midgut examination for oocysts and by enzyme-linked immunosorbent assay for P. falciparum circumsporozoite antigen. Calculations based on the age structure of the population and the resulting rates of mosquito infections indicated that children under 10 years of age were responsible for 72% of mosquito infections, individuals between 10 and 21 years of age contributed 12%, and those over 21 years of age accounted for 16%. No infection resulted in mosquitoes fed on infants less than 1 year of age.


Colonization factor different from K88, K99, F41 and 987P in enterotoxigenic Escherichia coli strains isolated from postweaning diarrhoea in pigs.

A novel common colonization factor was detected in enterotoxigenic strains of Escherichia coli isolated from intestinal contents of piglets affected with postweaning diarrhoea. This factor was antigenically distinct from the previously described K88, K99, F41, 987P, CFAI, CFAII and Att25 fimbrial antigens. E. coli strains possessing this factor adhered to the pig intestinal brush borders and one strain, used in experimental infection in weanlings, colonized the intestinal epithelium and induced diarrhoea. Examination of 212 toxigenic strains of E. coli isolated from weanlings revealed the presence of the novel common colonization factor in 83 strains, belonging to serogroups O25, O108, O138, O141, O147 and O157. The antigen K88 was detected in 47 strains belonging to serogroups O8, O141, O147 and O149.


The protective efficacy of pili from different strains of Moraxella bovis within the same serogroup against infectious bovine keratoconjunctivitis.

Three groups of ten calves were each immunised with a total of 400 micrograms pili prepared from three separate strains of Moraxella bovis in Alhydrogel-oil adjuvant as two divided, equal doses 21 days apart. Groups 1 and 2 each received a monovalent vaccine made from strain 4L and S276R respectively, which belonged to pili serogroup A. Group 3 received vaccine made from pili of strain Maff1, belonging to serogroup F. A further group of ten calves served as non-vaccinated controls. Calves in groups 1 and 2 had developed serogroup A-specific antibody and those in group 3 developed serogroup F-specific antibody, and some evidence of cross-reacting antibody was also detected when measured by an agglutination test using formalin-killed piliated cells of serogroup A strain 4L. Although antibody titres measured against purified pili by ELISA were highest with homologous serogroup antigens, cross-reactive titres to shared epitopes of M. bovis pili were also detected by this method. Ocular challenge of the 40 calves with virulent M. bovis of serogroup A strain S276R was carried out 14 days after the second vaccine dose. All non-vaccinated calves developed infectious bovine keratoconjunctivitis (IBK). The percentage protection in groups 1 (strain 4L) and 2 (strain S276R) was 60% and 80% respectively (P less than 0.05), with mean lesion scores of 0.7 and 0.3 out of a possible 6.0. The percentage protection of calves in group 3 (strain Maff1) was only 30%, with a mean lesion score of 1.4 compared with 2.2 for non-vaccinated controls. The present findings, together with other evidence indicating that immunity to IBK is serogroup-specific, suggest that inclusion of pili from one representative strain from each of the seven Australian and British serogroups in a polyvalent, subunit vaccine should effectively protect the majority of cattle against IBK caused by most field strains of M. bovis encountered in Australia and the United Kingdom.


Neuroendocrine carcinoma of the stomach with extensive somatostatin immunoreactivity.

Upper gastrointestinal tract neuroendocrine tumors producing predominantly somatostatin have thus far been described only in the duodenum; their characteristic features include the frequent presence of psammoma bodies (psammomatous somatostinomas), and the association with von Recklinghausen's neurofibromatosis. Gastric neuroendocrine tumors, on the other hand, tend to display immunoreactivity to serotonin but may include small subpopulations producing gastrin, motilin, pancreatic polypeptide, and somatostatin. In this report we describe a neuroendocrine carcinoma of the stomach with rapidly fatal outcome, displaying neurosecretory granules by electron microscopy and immunoreactivity to pan-neuroendocrine markers, ie, chromogranin and neuron-specific enolase. The only neuroendocrine regulatory peptide detected in the tumor was somatostatin, identified by immunohistochemistry in the majority of neoplastic cells. In contrast with duodenal somatostinomas, there were no psammoma bodies and no demonstrable association with von Recklinghausen's neurofibromatosis. To our knowledge this appears to be the first report of a malignant neuroendocrine tumor with diffuse somatostatin immunoreactivity.


Peroxisomes in human colon carcinomas. A cytochemical and biochemical study.

The presence of peroxisomes and their enzymic content were investigated and compared in healthy and neoplastic human colon epithelial cells using cytochemical studies at the ultrastructural level as well as biochemical analyses. Catalase-positive organelles were found to be more numerous in normal than in colonic neoplastic cells. Biochemical assays revealed that no D-aminoacid oxidase or L-alpha-hydroxyacid oxidase activity was detected in normal or tumor tissues. The specific activities of catalase, fatty-acyl CoA oxidase and enoyl-CoA hydratase/3 hydroxyacyl-CoA dehydrogenase (the so-called peroxisomal bifunctional enzyme of the beta-oxidation system) were found to be diminished in carcinoma cells compared with the control tissue. The fall in catalase activity correlated well with tumor stage according to Dukes, suggesting that this peroxisomal enzyme could be used as a potential prognostic marker.


Influence of synovial cells on cartilage in vitro: induction of breakdown and inhibition of synthesis.

Organoid or high density cultures of: (1) synovial cells from patients with rheumatoid arthritis, and (2) prechondrogenic mesenchymal cells from limb buds of 12-day-old mouse embryos, were co-cultured for 7-10 days using the Trowell culture system. Depending on the time of commencing co-cultivation, chondrogenesis was inhibited (co-cultivation from the start) or the cartilaginous matrix was partly degraded (co-cultivation after formation of embryonic cartilage, i.e. on day 4). These effects were obtained with cells from synovial fluid as well as from synovial tissue. Matrix degradation and the behaviour of the different cell types could be demonstrated well by electron microscopy under the in vitro conditions applied.


Tissue distribution of neutrophils in postischemic acute renal failure.

Polymorphonuclear neutrophil granulocytes (PMNs) seem to participate in the pathogenesis of renal ischemic reperfusion injury. The kidneys from male Sprague Dawley rats were immersion-fixed after 45 min of renal artery clamping followed by reperfusion for 0, 5, 20, and 120 min, respectively. The tissue distribution of PMNs in the kidneys was studied histochemically using naphthol AS-D chloroacetate esterase as a specific marker for these cells. Neutrophil counts per unit sectional area were obtained for renal cortex, outer and inner medulla. In the cortex separate intraglomerular and peritubular counts, and in the outer medulla separate outer and inner stripe counts were made. After 120 min of reperfusion the total renal PMN counts were 488 +/- 62 (n = 4) compared with 54 +/- 4 (n = 4) per cm2 in nonischemic controls. Within 120 min of reperfusion PMN counts increased by a factor of 8 in the cortex, of 12 in the outer medulla and of 14 in the inner medulla, compared with controls. The ratio of intraglomerular against peritubular PMN counts was approximately 2 in controls, but 0.5 after a 120-min reperfusion interval. The outer stripe of the outer medulla contained only a small number of PMNs whereas PMN counts of 923 +/- 197 (n = 4) per cm2 were found in the inner stripe after 120 min reperfusion. Interestingly, there was a marked increase in PMNs in the inner stripe during the first 5 min of reperfusion but no extravasation of PMNs was observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Intraperitoneal amyloid formation by amyloid enhancing factor--rich macrophages in ascitic fluid.

Although resident peritoneal cells from amyloidotic mice (amyloidotic peritoneal cells) are capable of processing the precursor protein of secondary amyloidosis, serum amyloid A (SAA) to amyloid fibrils, the peritoneum is a rare site for amyloid deposition. This is considered to be due to a deficiency of SAA in the peritoneum. To increase the supply of SAA to the peritoneum, ascitic fluid containing about the same protein constituents as in the serum was induced in mice. Amyloidotic peritoneal cells were packed in a microchamber which was shielded with filter membranes, and cultured in ascitic fluid supplemented with additional inflammatory factors. On the 7th day, Congo red-positive structures which showed green birefringence under polarized light were found inside and occasionally outside the chamber. By anti-AA or -SAA immunostaining, amyloid deposits and the cell surfaces of macrophages were positive. Immunologic depletion of T- and B-lymphocytes from the amyloidotic peritoneal cells did not adversely effect the amyloid formation in microchambers. These results suggest that either ascitic fluid containing sufficient amounts of SAA, or peritoneal macrophages with a high amyloid enhancing factor (AEF) activity are indispensable for AA amyloid fibrillogenesis in the peritoneum.


Altered transferrin gene expression in preneoplastic and neoplastic liver lesions induced in rats with N-nitrosomorpholine.

The expression of the gene for the iron transport protein transferrin was found to be altered in preneoplastic and neoplastic lesions induced in the rat liver by N-nitrosomorpholine. The total RNA of ten hepatocellular carcinomas (HCC) was investigated by Northern blot analysis using a cDNA-probe comprising 150 bp of the 3' region and compared with the total hepatic RNA in untreated rats. Seven hepatocellular carcinomas showed slight or pronounced reduction in transferrin expression. In situ hybridization of two additional hepatocellular carcinomas revealed marked reduction in the mRNA level for the transferrin gene compared with the surrounding tissue. In contrast, the majority of early preneoplastic lesions storing excess glycogen and tigroid cell foci expressed increased levels of transferrin mRNA. The loss of glycogen in mixed cell foci, which represent a later stage of hepatocarcinogenesis, was usually accompanied by a decrease in transferrin mRNA suggesting a close relationship between this change in gene expression and cellular dedifferentiation emerging during hepatocarcinogenesis.


Immunohistological study of skin involvement in Kikuchi's disease.

Five patients with histiocytic necrotizing lymphadenitis (Kikuchi's disease) with erythematous or popular skin lesions were studied. All patients healed naturally within a few months like the patients with no skin involvement. Histological findings for the skin lesions mimicked cutaneous malignant lymphoma. The infiltrated mononuclear cells usually demonstrated positive reactions for Ki-M1p (20-63%), lysozymes (13-54%), MT-1 (18-64%), UCHL-1 (22-39%) and LN2 (17-36%). OPD-4 and L26 positive cells were few in number. These results suggest that the infiltrated cells in a Kikuchi's disease skin lesion are composed of the same components as the affected lesion in the lymph node.


Low incidence of bronchus-associated lymphoid tissue (BALT) in chronically inflamed human lungs.

The relevance of bronchus-associated lymphoid tissue (BALT) in man is still under discussion. Animal experiments indicate that the development of BALT is dependent on microbial stimulation. Therefore, the incidence of BALT was investigated retrospectively in specimens removed during surgical procedures on patients with chronic pulmonary inflammation. All these patients had severe chronic bronchitis and bronchiectasis, but BALT was found in only 8%. In patients with BALT and a malignant tumor, occlusion of a bronchus with poststenotic pneumonia was always present and BALT was observed exclusively in areas peripheral to the occlusion. In man other compartments of the lung must be responsible for the immune function of BALT found in animals.


Pituitary lactotrophs and somatotrophs in pregnancy: a correlative in situ hybridization and immunocytochemical study.

Lactotroph hyperplasia is a prominent finding in the adenohypophyses of pregnant women. In order to elucidate the morphogenesis of this change, pituitaries from 16 women in various phases of pregnancy were collected at autopsy and studied by histology, immunocytochemistry and in situ hybridization. The results showed that the increase in the amount of prolactin (PRL) mRNA paralleled the progressive lactotroph hyperplasia. The presence of mitoses in PRL-immunoreactive cells provided evidence that proliferation of preexisting lactotrophs contribute to lactotroph accumulation. Growth hormone (GH) immunoreactive cells showed a marked reduction in GH mRNA indicating that GH synthesis was inhibited. In many GH-immunoreactive cells, PRL mRNA became apparent. These findings demonstrate that GH is stored following discontinuation of GH synthesis. It appears that, when PRL is secreted in excess during pregnancy, somatotrophs are recruited to produce PRL. These somatotrophs begin to express PRL mRNA, transform to bihormonal mammosomatotrophs and possibly later to lactotrophs, contributing to PRL production. Mature somatotrophs may be regarded as reserve cells in the adenohypophysis, having the potential to switch hormone synthesis and to convert to mammosomatotrophs and possibly lactotrophs.


Analysis of proliferative activity in colorectal mucosa by immunohistochemical detection of proliferating cell nuclear antigen (PCNA). Methodological aspects and application to routine diagnostic material.

Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) has been suggested as a new approach for determining proliferative activity in paraffin-embedded tissue. In a prospective study PCNA immunostaining was performed in 284 colorectal biopsies using monoclonal antibodies 19F4 (Ogata et al. 1987) and PC10 (Waseem and Lane 1990) and compared with the Ki67 method. From each site three biopsies were taken and a variety of fixation regimens for frozen and paraffin-embedded samples tested. For frozen biopsies methanol fixation at -20 degrees C proved best. In paraffin sections PCNA could be detected after methacarn fixation as well as after controled fixation at 4 degrees C in 4% paraformaldehyde for 1 h and in most biopsies routinely fixed with 10% formalin. However, the latter fixation regimens revealed additional PCNA-positive cells in the normal superficial colonic mucosal epithelium. Although the percentage of cells positive for PCNA was generally lower than for Ki67, the rates correlated in a highly significant fashion, both in frozen methanol-fixed biopsies, and in paraformaldehyde-fixed paraffin-embedded samples. PCNA immunohistochemistry revealed a similar proliferative activity in different parts of the large bowel. A higher proliferative activity was found in inflamed mucosa, adenomas, carcinomas and even in normal mucosa from patients with colorectal neoplasms. In routinely fixed biopsies, the monoclonal antibody PC10 was superior to 19F4 because of considerably less background staining. However, in the routine material only a rough estimate of the proliferative activity was possible by PCNA immunohistochemistry using these antibodies, because unpredictable numbers of non-S-phase cells were also stained.(ABSTRACT TRUNCATED AT 250 WORDS)


Interphasic nucleolar organizer regions expression and cell kinetics evaluation during gastric carcinogenesis induced by nitrosoguanidine in the rat.

An increased number of interphasic nucleolar organizer regions containing ribosomal cistrons associated with argyrophilic proteins (AgNORs) has been described in human malignant tumor cells. In this study variations in AgNOR numbers have been compared with changes of cell kinetics, evaluated by the mitotic count (MC) and bromodeoxyuridine labeling index (BrdU LI), during gastric carcinogenesis induced with N-methyl-N'-nitro-N-nitrosoguanidine (NG) in rats. Significant differences (2 P < 0.005) in AgNOR mean numbers, evaluated in the antral isthmic cells, in MC mean values and BrdU LI, evaluated in the whole antral cellular population, were found when comparing areas of acute gastritis, atrophy and hyperplasia in NG-treated rats with the normal mucosa in controls. No differences were observed in MC and BrdU LI between normal antrum and carcinoma cells which showed an AgNORs mean number lower than in the isthmic cells of controls (2 P < 0.005). Moreover, significant correlations were found comparing changes in AgNOR numbers with MC (r = 0.89, P < 0.001) and BrdU LI (r = 0.66, P < 0.001) in different lesions. These data show that evaluation of AgNOR numbers does not allow the identification of malignant cells in NG-induced gastric carcinoma. However AgNOR quantification seems to be a reliable index of cell kinetics and related well with the cellular dividing fraction.


NCI-H716 cells as a model for endocrine differentiation in colorectal cancer.

In colonic neoplasms, endocrine differentiation is encountered not only in carcinoid tumors but also in adenocarcinomas, where endocrine cells may represent a distinct line of differentiation in the tumor. The significance of endocrine differentiation in colorectal cancer is not well established, partly because of the paucity of tumor cell lines which can serve as a model for studying endocrine differentiation. In this report we describe the properties of NCI-H716 cells, a cell line derived from a poorly differentiated adenocarcinoma of the caecum, under various in vitro conditions and as xenografts in athymic mice. Phenotypical properties were immunohistochemically assessed using a panel of differentiation related antibodies, and also by Northern blot analysis and by electron microscopy. Receptors for biogenic amines and peptide hormones were analyzed by ligand binding assay. These studies show that: 1. NCI-H716 cells can be undifferentiated, or show endocrine, mucin-producing or "amphicrine" properties. 2. Endocrine differentiation of NCI-H716 cells preferentially occurs in xenografts in athymic mice, which suggests that mesenchymal elements induce endocrine differentiation. 3. NCI-H716 cells express large amounts of high affinity receptors for gastrin, serotonin and somatostatin and these substances can regulate growth. Thus, NCI-H716 cells form a suitable model for the study of endocrine differentiation in intestinal epithelium and of auto- or paracrine growth regulation in intestinal neoplasia.


[A trial of the pharmacological correction of sleep disorders in sailors during a cruise].

The article makes a comparative estimation of the efficiency between the benzodiazepine pharmacological group (selenium, diazepam) and adaptogens of the plant origin (Extractum Leuzeae carthamoidis fluidum) for correction of sleeping disorders in seamen during cruise. The application of elenium and diazepam have caused the unpleasant subjective sensations, such as psychoemotional retardation, sluggishness, apathism and headaches. After application of Extractum Leuzeae there were no side effects like a drug hang-over; the health status, functional state and job performance of seamen were improved. Having taken Extractum Leuzeae the seamen had the normalization of sleeping in 5-7 days.


[The characteristics of the pharmacotherapy for patients with ischemic heart disease and hypertension in relation to age].

On the basis of literary and own findings the authors discuss disputable problems of the pharmacotherapy of ischemic heart disease and hypertensive disease in the age aspect including patients over 60 years of age. Approaches are analyzed to individual choice of adequate pharmacotherapy, criteria of selection and dosages with special emphasis of adreno-beta-blockaders and calcium antagonists as well as criteria of evaluation of pharmacotherapy.


[A method for the permanent maintainance aurotherapy of rheumatoid arthritis patients under the control of gold excretion].

A study is presented of 34 patients with rheumatoid arthritis. The authors propose a method of persistently maintained aurotherapy of patients with rheumatoid arthritis directed to prevention of recurrences and toxic effect of gold due to chrysanol treatment, the active substance of which is metallic gold. After course treatment with chrysanol (850-1020 mg) persistent treatment by maintained doses is instituted. prevention of recurrences of the disease and prevention of side effects due to the toxicity of cumulated gold.


[Effect of physiologically active polymers on the activity and hormonal induction of tyrosine aminotransferase].

Effect of neutral polymers on activity of tyrosine aminotransferase and hydrocortisone induction of the enzyme were studied. Physiologically active polymers decreased in most cases the basal rate of the enzymatic activity. Preadministration of polymers decreased hormonal induction of tyrosine aminotransferase. Simultaneous administration of polymers and hydrocortisone led to augmented and long-term induction, which appears to occur due to complex-formation of the polymer and hormone. Gradual liberation of hydrocortisone from the complex enabled to maintain the hormone effective concentration for a long time.


[The question of amino acid isomerization in the microwave field. Results of a model study with standard solutions].

Aqueous standard-solutions of L-alanine, L-glutamic acid, and L-proline do not reveal any increase of D-enantiomers after 30 min heating--neither by the conventional method on a hotplate, nor in a standard microwave oven. A specific "microwave effect" and, hence, a special consumer risk is, in contrast to recent assumptions, not detectable. Effects on the amino acids which were observed in conventionally heated samples are explained by higher heat-exposure during the treatment of these samples.


[Validation of the Child Speech Test using the Heidelberg Speech Development Test with six-year-old preschool children].

We present and discuss results from researches to validate a new test for diagnosing speech-competences of preschool children KISTE. 41 children (6;0-6;11) were tested both with KISTE and HSET. Correlations, factor- and clusteranalysis showed that the structure of both tests is different and that KISTE has a good differential validity for both aspects of speech: communicative and structural.


[The disposition of natural foci of hemorrhagic fever with renal syndrome in different landscape areas of Tyumen Province].

Lungs of 3159 animals of the forest complex from 90 areas of 30 administrative districts of Tyumen Province were examined by enzyme immunoassays for antigen of hemorrhagic fever with renal syndrome (HFRS) during 5 years, 1985-1989. The antigen of HERS virus was detected in the lungs of mammals of 8 species: Clethrionomys glareolus and Cl. rutilus, Siberian and Arctic lemmings (first findings in the world), M. oeconomus, field mouse, common and pygmy shrews. Nearly all the findings refer to the subzone of southern taiga and adjacent areas of subtaiga subzone where Cl. glareolus is the main reservoir of infection and Cl. rutilus an additional one. In the tundra zone, Siberian lemming is the main reservoir of infection and Arctic lemming an additional one. No natural foci of HFRS were found in forest steppe and forest tundra zones. In the subzone of the northern and middle taiga, the antigen was found only on 4 occasions: 3 in common shrews and one in Cl. glareolus (near the town of Khanty-Mansisk). An irregular annual infection rate with HFRS virus was observed in Cl. glareolus as well as its decline from spring to autumn. It cannot be ruled out that lemmings are carriers of a distinct HFRS virus serotype.


[Pharmacology of excitatory amino acid receptors].

L-Glutamate is an excitatory neurotransmitter at many synapses in the animal, and causes neuronal damage in the mammalian neuron. Glutamate receptors have been classified into at least five types: NMDA-, kainate-, AMPA-, and L-AP4-type ionotropic receptors and metabotropic glutamate receptors. Recently new glutamate agonists have been introduced into the pharmacological study in neuroscience research. They are methoxyphenylkainoids, acromelic acids and 2-(carboxycyclopropyl)glycine (CCG). A methoxyphenylkainoid and acromelic acid A are one of the most potent excitants in the mammalian central neuron. These compounds should provide valuable pharmacological probes for analysis of functions of excitatory amino acids.


[The intensity of the immune response to the hemorrhagic fever with renal syndrome virus among different population groups of Bashkortostan].

For the first time the natural immune stratum to hemorrhagic fever with the renal syndrome (HFRS) among the population of all regions of Bashkiria, a large territory with the highest morbidity level in this infection in the USSR, was studied. 9,176 persons over 15 years of age were examined by radioimmune techniques. Analysis of the immune structure of the population revealed that the share of immune males was higher than that of immune females, but the difference was less than the difference between males and females among the registered HFRS patients. Among children immune persons were almost completely absent. The immune stratum increased rather slowly among persons under 40 years, and only from this age the considerable increase of the immune stratum was observed (up to 16.2%). High risk groups included persons of such professions as forestry workers, truck and tractor drivers, oil workers and prospectors, livestock breeders, builders. These data may be used for the formation of groups to be vaccinated against HFRS in foci with different degrees of activity. Before vaccination the correction of groups to be vaccinated in a given region should be carried out, taking into account the immune structure of the population and the specific features of the local landscape and economy.


[Drug therapy of peptic ulcer in the elderly].

Medical therapy of peptic ulcers in elderly patients should depend on drugs which have to be administered only once daily. In addition, the ideal compound should have minimal side effects as well as minimal, if any, interactions with concomitant forms of therapy. From this point of view, acid-inhibitory drugs are preferable to other anti-ulcer drugs, the latter having little importance in advanced age. For ulcer healing, the proton pump inhibitor omeprazole is recommended, whereas H2-blockers (predominantly ranitidine) are the first choice in long-term therapy in order to prevent relapses. Treatments directed towards eradication of Helicobacter pylori should be regarded with great caution in the elderly at the present time.


[The use of the bromocriptine test in schizophrenia patients resistant to neuroleptic therapy].

To choose differentiated therapy, 62 patients suffering from refractory schizophrenia were administered bromocriptine in a dose of 7.5-15 mg/day after they had been placed on placebo. According to the bromocriptine test, the treatment was changed in 53 patients. After the occurrence of the picture of hallucinatory stupor remission was attained with the aid of less doses of neuroleptics given before. In patients who responded to bromocriptine by anxious excitation, the health status noticeably improved after administration of anxiolytics and lithium carbonate. Those patients in whom bromocriptine eliminated psychosis were treated later with the drugs belonging to the group of monosubstituted benzamides.


Attenuated ventilatory response to hypoxaemia at vecuronium-induced partial neuromuscular block.

The effect of a partial neuromuscular block on the ventilatory response to hypercarbia and to hypoxaemia was studied in 11 non-anaesthetized male subjects. Respiratory frequency, tidal volume, minute volume, respiratory timing and drive were measured during air breathing and during stimulation by hypercarbia and hypoxaemia. The ventilatory response was defined as the ratio between, respectively, tidal volume and minute volume during ventilation stimulated by hypercarbia and hypoxaemia compared to measurements during air breathing. The ventilatory measurements were repeated on three separate occasions: before neuromuscular block was established, during an infusion of vecuronium aiming at a mechanical adductor pollicis train-of-four (TOF) ratio of 0.70, and after the infusion had been stopped and the neuromuscular block had spontaneously recovered to a TOF ratio of > 0.90. Resting ventilation during air breathing remained with minor variations throughout the experiment. The ventilatory response to hypercarbia was not affected at a TOF ratio of 0.70 as compared to measurements before vecuronium and at a TOF ratio of > 0.90. In contrast, the ventilatory response to hypoxaemia was markedly reduced at a TOF ratio of 0.70. We conclude that a mechanical TOF ratio of 0.70 following vecuronium may be associated with an inadequate ventilatory response to hypoxaemia.


Creutzfeldt-Jakob disease with codon 129 polymorphism (valine): a comparative study of patients with codon 102 point mutation or without mutations.

We examined 7 patients with Creutzfeldt-Jakob disease (CJD) with a methionine-to-valine change at prion protein (PrP) codon 129 (CJD129 patients). These CJD129 patients did not have either a codon 117 or 198 point mutation. For comparison, we also examined 7 patients with Gerstmann-Sträussler syndrome (GSS) with a proline-to-leucine change at PrP codon 102 (GSS102 patients) and 13 patients without any known mutations at codons 102, 117, 129, 178, or 200 (CJDwild patients). CJD129 patients had a long clinical duration and ataxia at onset, but rarely had any periodic synchronous discharge in their electroencephalogram. Unlike CJDwild patients, all CJD129 patients have typical congophilic PrP plaques in their brain. These clinicopathological findings were similar to those of GSS102. However, the distribution and morphology of PrP deposits revealed by immunohistochemistry were different between CJD129 and GSS102. In GSS102 more numerous and various types of PrP plaques are seen throughout the brain, while in CJD129 patients a unicentric core was the major feature of PrP plaques. The change in codon 129 influences the clinical course and pathological findings in CJD.


Doppler sonographic pulse curve analysis for estimation of the haemodynamic function.

Experience with Doppler sonographic pulse curve analysis is given from 40 glaucoma patients (examination of the temporal posterior ciliary artery), and from premature babies (blood flow in the anterior cerebral artery). In the latter group the reaction to pain and stress connected with blood sampling was studied. In the glaucoma patients focus was on the influence of IOP level on vascular autoregulation, and the possible response to therapy with beta-blockers. Describing an averaging technique, methodological problems are discussed as relevant for the detection of changes in flow and velocity in small blood vessels.


The statoconial membrane of the guinea pig utricular macula. Scanning electron microscopic investigation combined with the freeze-fracturing technique.

The superstructure of the guinea pig utricular macula was investigated using scanning electron microscopy combined with the freeze-fracturing technique. The statoconial membrane was composed of the otoconial layer, otolithic membrane and subcupular meshwork. The otolithic membrane consisted of closely arranged filaments with beaded appearance and densely packed globular matrix near the otoconial layer. The subcupular meshwork consisted of long branching filaments cross-bridged to one another. The filaments were continuous with those of the otolithic membrane on one side and with the surface of the epithelium on the other, which fills the space between the otolithic membrane and the macular surface. The otolithic membrane would function as a rigid plate and equally distribute the shear force of the otoconial layer to all the ciliary bundles. The subcupular meshwork may play an important role in transmitting the shear strain of the otolithic membrane to all the ciliary bundles and may also exert an additional dampening effect to prevent unwanted vibration.


[Synthesis, analgesic activity and structure-activity relationship of 4-N-propionyl analogs of cis-3-methyl fentanyl].

The synthesis of some 4-N-propionyl analogs of cis-3-methyl fentanyl by acyl chloride method. anhydride method, mixed anhydride method and DCC method is reported. The ethyl group in 4-N-propionyl portion of cis-3-methyl fentanyl was substituted by some groups with different electronic property. In mouse hot plate test, all compounds showed typical morphine-like action with analgesic potency corresponding to 49-1963 times that of morphine. Compound 3 was found to exhibit higher analgesic activity than cis-3-methyl fentanyl. Semiempirical INDO calculations have been undertaken for 4 typical compounds and it was found that as a result of introduction of chlorovinyl, compound 3 exhibited characteristics of electronic structure different from that of cis-3-methyl fentanyl. The relationships between analgesic activity and the electronic structure of these compounds were discussed.


Effects of picroliv, the active principle of Picrorhiza kurroa, on biochemical changes in rat liver poisoned by Amanita phalloides.

The efficacy of Picroliv, a standardized iridoid glycoside fraction of Picrorhiza kurroa, was studied against the Amanita phalloides-induced biochemical changes in rat liver. A phalloides (50 mg.kg-1) caused significant increases in the activities of hepatic 5'-nucleotidase, gamma-glutamyl transpeptidase, acid ribonuclease, and succinate dehydrogenase, but a decrease in glucose-6-phosphatase. The level of cytochrome P-450 in microsomal fraction and content of glycogen in liver showed significant depletions. Picroliv (25 mg.kg-1.d-1 x 10 d) provided significant restorations of all the biochemical changes poisoned by A phalloides except cytochrome P-450 and glycogen. These results demonstrated the protective effect of Picroliv against A phalloides-induced hepatotoxicity in rats.


[Teratologic studies of methyl 5-aminosalicylate and salicylazopyridine].

Methyl 5-aminosalicylate hydrochloride (M-5-AS) at the dosages of 21, 42, 209, and 417 mg.kg-1.d-1 ig to mice during d 6-15 of pregnancy, no obvious effects on the placenta, fetus weight, sex differentiation, external appearance, visceral, and skeletal development were observed. In rats ig M-5-AS 56 and 556 mg.kg-1.d-1 produced no noticeable effects on the organogenesis or teratogenesis either. In mice and rats ig salicylazopyridine 410 and 1089 mg.kg-1.d-1 respectively no obvious teratogenicity was detected. However, aspirin 250 mg.kg-1.d-1 ig did bring about significant teratogenicity in rats.


Adrenergic and non-adrenergic cardiovascular effects of thyrotropin-releasing hormone (TRH) in the anaesthetized rabbit.

The effects of thyrotropin-releasing hormone (TRH) on regional blood flows were studied in urethane-anaesthetized rabbits. Experiments were performed both with and without adrenergic antagonist pretreatment. The tracer microsphere method was used to measure blood flow. TRH (0.1 mg kg-1) caused an increase in mean arterial blood pressure (MAP) from 9.8 +/- 1 to 11.8 +/- 0.8; a higher dose (1 mg kg-1) increased the blood pressure to 15.2 +/- 1 kPa (P less than 0.001). Total cerebral blood flow (CBFtot) increased to 137 +/- 10% (P less than 0.05) of control at the lower dose and to 214 +/- 16% (P less than 0.001), at the higher dose. A reduction in blood flow at both doses of TRH in several peripheral organs indicates that the pressor effect was mainly due to an effect on the peripheral vascular resistance. In prazosin-pretreated animals in which the MAP was normalized by ligation of the thoracic aorta, TRH elicited an increase in the CBFtot to 131 +/- 12% (P less than 0.05) of control. In the iris, TRH caused vasodilation in prazosin-pretreated animals. In experiments with combined alpha- and beta-adrenergic blockade, a non-adrenergic vasoconstricting effect of TRH was seen in some peripheral organs. The results indicate that TRH activates the sympathetic nervous system thus causing an increased vascular resistance and MAP; these effects are mediated mainly by an alpha 1-adrenergic mechanism. In the spleen, the gastric mucosa and the adrenal glands, the vasoconstriction caused by TRH was partly non-adrenergic. The vasodilation seen in the small intestine and the anterior uvea after TRH treatment and adrenoceptor blockade may be explained by effects on the parasympathetic nervous system. The vasodilating effect of TRH in the brain does not seem to involve alpha 1- or beta-adrenergic mechanisms.


Effects of somatostatin on glucagon-stimulated glycogenolysis and gluconeogenesis in hepatocytes cultured in vitro.

The effects of somatostatin (SS-14) on glycogenolysis and gluconeogenesis in rat hepatocytes cultured in vitro in a serum-free medium were investigated. Somatostatin (122 nmol l-1) did not significantly change the basal glucose production with or without pyruvate (10 mmol l-1). Glucagon strongly (over 100%) increased the glucose production in hepatocytes incubated in a medium supplemented with 10 mmol l-1 pyruvate. This increase in glucose production is the result of increased rates of gluconeogenesis and glycogenolysis. Somatostatin partially inhibited the glucagon stimulated increase in glucose production. Glucagon also significantly increased the glucose production in a glucose-free medium without pyruvate, which resulted from an increase of glycogenolysis. Somatostatin did not inhibit the increase in glucose production in these conditions. After a 4 h 'fast', glycogen in hepatocytes fell to a very low level. Glucose production was minimal. After the addition of pyruvate, there was a increase in gluconeogenesis and glucose production. Glucagon stimulated the rate of gluconeogenesis. Somatostatin completely inhibited this glucagon-stimulated increase in gluconeogenesis.


Effect of gamma-glutamyl transpeptidase inhibitors on the transport of glutamate into neuronal and glial primary cultures.

Inhibitors of gamma-glutamyl transpeptidase (mixture of serine and borate--13 mM, kainic acid--5 mM and 6-diazo-5-oxo-L-norleucine--2 mM) significantly suppressed glutamate uptake into cultured neurones and glial cells. The simultaneous application of any of these inhibitors with ouabain resulted in a further decline in glutamate uptake. It can be speculated that gamma-glutamyl transpeptidase significantly contributes to glutamate transport into nerve cells in the early period of brain development until the Na(+)-K(+)-gradient is fully constituted.


Heparin binding properties of the carboxyl terminal domain of [A103,106,108] antistasin 93-119.

Antistasin is a 119 amino acid protein with anticoagulant, antimetastatic and heparin-binding properties derived from the salivary glands of the leech Haementaria officinalis (1). This protein contains a specific consensus sequence for heparin binding at its carboxyl terminal end and a region between residues 32 and 48 putatively involved in glycosaminoglycan interactions. The cyclic peptide antistasin 37-48 (C-P-H-G-F-Q-R-S-R-Y-G-C) and the carboxyl terminal fragment [A103,106,108] antistasin 93-119 (P-N-G-L-K-R-D-K-L-G-A-E-Y-A-E-A-R-P-K-R-K-L-I-P-R-L-S) were synthesized by solid-phase peptide chemistry and their interactions with 125I-labeled heparin were investigated. Heparin binding to [A103,106,108] antistasin 93-119 was specific and saturable as binding was blocked by addition of the unlabeled glycosaminoglycan. The rank order of potency of various glycosaminoglycans in blocking 125I-labeled heparin binding to [A103,106,108] antistasin 93-119 was dextran sulfate greater than heparin much greater than dermatan sulfate greater than or equal to chondroitin sulfate A and C indicating a specificity of the peptide for the glycosaminoglycan structure. Moreover, heparin binding increased linearly with increasing salt and was optimal at 0.15 M NaCl and physiological pH. In contrast, binding of heparin to the basic peptide antistasin 37-48 decreased linearly as the ionic strength of the medium was increased to physiological concentration (0.15 M) thus showing a greater specificity of heparin for [A103,106,108] antistasin 93-119. These studies indicate that residues 93-119 of antistasin mediate this inhibitor's interaction with heparin.