Endothelium-dependent regulation of resistance arteries: alterations with aging and hypertension.

Small arteries with a diameter of 200 microns or less play an important role in the regulation of peripheral vascular resistance. Dysregulation of vascular tone of these arteries may contribute significantly to high blood pressure. The contractile state of blood vessels is regulated by peripheral neurons, vascular smooth muscle, and the endothelium. In perfused mesenteric resistance arteries of the rat, removal of the endothelium markedly augments the sensitivity and maximal response to norepinephrine and endothelin-1. Acetylcholine causes profound endothelium-dependent relaxation of resistance arteries contracted with norepinephrine or endothelin-1. The potency of the muscarinic agonist is particularly pronounced with intraluminal application. The inhibitory effects of the endothelium against contractions induced by norepinephrine and endothelin-1 are reduced with aging and hypertension. The endothelium-dependent relaxation in response to intraluminal, but not extraluminal, acetylcholine is blunted in mesenteric resistance arteries of hypertensive rats and in the forearm circulation of hypertensive patients studied in vivo. The sensitivity of vascular smooth muscle to the effects of endothelin decreases with advancing age and in the spontaneously hypertensive rat. Thus, endothelium-derived vasoactive substances can profoundly affect vascular tone of resistance arteries studied in vitro and in vivo. The inhibitory effects of the endothelium against vasoconstrictor stimuli decreases with aging and hypertension, indicating a dysfunction of these regulatory mechanisms under these conditions.


Determining hypertensive end-organ damage in trials: a review of current methodologies and techniques.

The determination of hypertensive end-organ damage is the most crucial aspect of any long-term hypertension trial. Definition of inclusion criteria and withdrawal rules, and the evaluation of the treatment regimen in light of the cardiovascular prognosis, are all largely dependent on the trialists' abilities to identify vascular lesions and/or functional deteriorations of the major target organs. Although the vascular complications of hypertension tend to be diffuse, some organs and tissues are particularly sensitive: brain, eye fundus, heart, conduit arteries, and kidneys. To achieve a modicum of diagnostic accuracy, trial investigators will have to combine clinical acumen with a selection of ancillary techniques. This article reviews the compromises resulting from weighing the available technological potential against such conflicting factors as convenience and compliance of trial subjects, and expenditure.


The molecular basis of cardiovascular hypertrophy: the role of the renin-angiotensin system.

Factors that can influence cardiovascular growth are becoming increasingly important for our understanding of such complex diseases as cardiac hypertrophy, coronary artery disease, atherosclerosis, and hypertension. Several proto-oncogenes were found to be involved in the regulation of abnormal cell growth in cardiovascular disease. It is also evident that some peptide hormones, which are well known to be involved in blood pressure control, may play a role as growth modulators. Angiotensin II is one such peptide. It elevates blood pressure through its direct vasoconstrictor, sympathomimetic, and (through release of aldosterone) sodium-retaining activity but also appears to have mitogenic actions. Interestingly, all components of the renin-angiotensin system were found locally in cardiovascular tissues. The question remains whether angiotensin can act directly as a growth factor or whether it does so indirectly by influencing or modulating cell growth factors. A better understanding of the renin-angiotensin system as a direct or indirect mediator for cardiovascular hypertrophy would offer new and interesting insights into the pathophysiology of hypertension and possibly novel options for the treatment of cardiovascular disease.


Hypertensive heart disease: pathophysiology and clinical and prognostic consequences.

The determinants of cardiac output were investigated in 161 patients with essential hypertension World Health Organization (WHO) stages I and II. In multiple regression analysis, cardiac output was inversely and independently related to blood pressure and to age. In patients with more severe hypertension, the lower cardiac output was associated with a lower stroke volume and a higher peripheral oxygen extraction. When age and blood pressure were taken into account, cardiac output was not a significant predictor of total mortality and of future cardiovascular events. Clinic and casual blood pressure explain only up to about 30% of the variability of echocardiographic left ventricular mass. The relationship of electrocardiographic voltages with blood pressure at various levels of bicycle exercise was highly significant in 169 patients with essential hypertension (r = 0.29-0.38; p less than 0.001) but the relationship was not better than with pressure at rest (r = 0.39). Ambulatory blood pressure, however, may be better related to left ventricular mass than clinic pressure. Several studies indicate that left ventricular hypertrophy is a significant risk factor for future cardiovascular events, independent of age and blood pressure. Left ventricular systolic function is usually normal in established hypertension, but diastolic function is frequently impaired, which could explain the reduced peak oxygen uptake in patients with more severe hypertension.


The concept of coronary flow reserve.

Coronary reserve has been defined as the ratio of coronary resistance under control (rest) conditions and of coronary resistance after maximal coronary vasodilation. The latter can be achieved by various interventions, the most important and clinically relevant example being intravenous administration of dipyridamole at 0.5 mg/kg of body weight. For patients without coronary artery disease, the coronary reserve is about 400 to 500%, i.e., the normal heart is capable of reducing its coronary resistance to minimal values of 0.18 to 0.2 mm Hg/ml/min/100 g or to increase coronary flow by approximately four- to fivefold. The determination of coronary reserve in humans implies the availability of adequate methods. Systematic analyses of different coronary blood flow measurements have proved the gas chromatographic argon method to be the most appropriate and accurate method for clinical conditions, as previously described in detail. In this report, our findings on the coronary reserve analysis in various clinical conditions are described as follows: (a) coronary artery disease, (b) inflammatory disturbances of the microcirculation, (c) hypertensive microangiopathy, (d) rheologic abnormalities of the heart, and (e) pressure and volume overload due to hypertension and heart valve lesions (metabolic overload).


Hypertension and vascular disease.

The use of antihypertensive drug treatment has altered the natural history of hypertension. Whereas congestive heart failure, cerebral hemorrhage, and renal failure were the major complications of untreated severe hypertension, myocardial infarction and thrombotic stroke have emerged as major problems in treated hypertensive patients. None of the major therapeutic trials in hypertension have provided evidence that reducing blood pressure reduces the risk of atherosclerotic complications of hypertension. Hypertension certainly aggravates the severity of atheromatous lesions in experimental animals, and may do so in humans. However, atherosclerosis is more closely related to disturbances in lipoprotein metabolism than to other factors. The common finding that serum cholesterol is raised in hypertensive patients may be due to atherosclerosis being the primary lesion, and hypertension a secondary complication rather than hypertension being the primary lesion.


Diagnosis of left ventricular hypertrophy by echocardiography.

Left ventricular hypertrophy in systemic arterial hypertension is associated with an increased risk of morbidity and mortality due to cardiovascular disease. Therefore, the diagnosis of left ventricular hypertrophy is clinically important. In current clinical practice, echocardiography is the method of choice for diagnosing left ventricular hypertrophy. This review describes current, clinically applied techniques of measuring left ventricular mass using M-mode and two-dimensional echocardiography. Using M-mode techniques, left ventricular hypertrophy is usually present when myocardial mass estimates exceed 150 g/m2 in males and 120 g/m2 in females. Using two-dimensional echocardiography, upper limits of normal have been described to be slightly lower (102 g/m2 in males and 88 g/m2 in females). In serial clinical two-dimensional echocardiographic studies, image acquisition and quantitation predominantly determine total variability in left ventricular mass estimates. Using any single technician and any single reader, left ventricular mass estimates in normal subjects may vary by 35 g (standard deviation) between serial studies. Strategies to reduce the magnitude of this variability include increasing the number of technicians and readers acquiring and analyzing a single study.


Studies of the heart using magnetic resonance.

Cardiovascular medicine is based on high-technology management of end-stage disease. Preventive medicine is preferable but, before magnetic resonance (MR), there was no noninvasive, safe, repeatable method of detecting occlusive vascular disease, which accounts for more deaths than any other disease at a presymptomatic stage. Demographic and animal studies show that occlusive vascular disease is not a normal aging process and is reversible. There was no method of monitoring the efficacy of preventive and therapeutic measures other than long, expensive, and sometimes inconclusive clinical trials. The rate of precession of atomic nuclei in a magnetic field is related to field strength. Creating a unique magnetic field in each volume of interest allows analysis of emitted radio signals to produce an image containing information about movement and local biochemical environments. MR produces accurate high resolution anatomical images and functional information including blood flow. It is already established as a method of choice in clinical cardiology for congenital heart disease and aortic disease. It is becoming established as a method of evaluating cardiovascular-active drugs, and in the future MR will be used for diagnosis and as a population-screening instrument for all cardiovascular disease including coronary artery disease.


Prognostic significance of micrometastatic tumour cells in bone marrow of colorectal cancer patients.

Much of the information recorded at the time of surgery for malignant disease has been used, alone or in combination, to indicate the outlook for that patient, including probability of metastasis. However, direct evidence of tumour seeding in distant organs at that time is not available. An immunocytochemical assay for the epithelial cytokeratin protein (CK18) may fill this gap since it is a feature of epithelial cells but would not normally be in bone marrow. We looked for disseminated tumour cells preoperatively in bone marrow from 88 patients with radically resected colorectal carcinomas. Smears for 28 (32%) patients were positive for cells of epithelial origin, which were absent from aspirates taken from 102 controls with non-malignant disease. The prognostic value was assessed in a follow-up study with a median observation time of 35 (12-58) months. Patients who had bone marrow tumour cells in the aspirate showed a significantly shorter disease-free survival than those without tumour cells (p = 0.0084). In a Cox regression model multivariate analysis demonstrated that the finding of tumour cells in bone marrow is an independent, significant (p = 0.0035) determinant of relapse. Although the skeleton is not a preferred site of overt metastasis in colorectal cancer the demonstration of tumour cells in bone marrow has to be taken as evidence of the general disseminative capability of an individual tumour.


Ganglioside-induced inhibition of in vivo immune response in mice.

In vitro immunomodulatory properties of gangliosides have been well characterized such as the ganglioside-induced modulation of CD4 on T lymphocytes and inhibition of lectin-induced proliferative response of lymphocytes. These findings have led to an interesting suggestion that gangliosides play a role as in vivo immune modulators, although this possibility is not clearly defined yet. We then first confirmed in vitro effects of gangliosides on murine immunocytes and examined in vivo effects of gangliosides on immune response in mice. Murine spleen cells that were treated with a ganglioside mixture (GS) purified from bovine brain exhibited a marked decrease in CD4 expression, while CD8 expression was slightly suppressed. Transplantation of GS-untreated control immunocytes that were isolated from syngeneic mice into the immune suppressed mice by X-ray irradiation restored in vivo immune responses, while GS-treated cells could not. Immune response was assayed by the evaluation of footpad swelling which was induced by immunization with sheep erythrocytes as antigens. Moreover, intramuscular administration of gangliosides into mice suppressed both immediate (Arthus)-type and delayed-type allergic reactions. These results suggest that gangliosides would be potential in vivo immune modulators.


Site-specific antibodies to the DNA-binding domain of estrogen receptor distinguish this protein from the 3H-estradiol-binding protein in pancreas.

The estradiol-binding protein (EBP) in extracts of rat and rabbit pancreata was characterized by sucrose density gradient analysis, immunoaffinity adsorption and Western blot (immunoblot) analysis using polyclonal antibodies raised against EBP. Rat pancreatic extracts labeled with 3H-estradiol contained a readily resolvable peak of steroid-binding activity that sedimented as a 4S complex on sucrose density gradients in the presence or absence of 0.4 M KCl. Estrogen receptor (ER) from calf uterine cytosols sedimented as a 4S complex on gradients containing 0.4 M KCl and as an 8S entity on gradients without KCl. Incubation of cytosol fractions from rat pancreas and calf uterus with benzoyl-DL-arginyl-p-nitroanilide (BAN) increased specific binding of 3H-estradiol to EBP but not to ER. Furthermore, two distinct site-specific antibodies to the DNA-binding domain of ER caused a marked increase in sedimentation rate of 3H-estradiol-labeled ER while normal rabbit serum and antibodies against EBP were ineffective in this regard. These data suggest that a significant portion, if not all, of the DNA-binding domain of ER is absent from EBP. Examination of the amino acid sequence of the DNA-binding domain of ER revealed a region of 10 amino acids that is significantly homologous to somatostatin, a tetradecapeptide that is a co-ligand in the binding of 3H-estradiol to EBP. Based on this observation, a possible mode of action of EBP in pancreatic acinar cells is proposed.


Induction of a humoral immune response to a Shiga toxin B subunit epitope expressed as a chimeric LamB protein in a Shigella flexneri live vaccine strain.

Shigella flexneri vaccine strain (SFL124) given orally, evokes humoral immune response in human volunteers. Such a strain, expressing antigenic epitope of B subunit of Shiga toxin, would also provide immunity to the toxin produced by some species of Shigella. A synthetic oligonucleotide, specifying an epitope [13-26 amino acids (aa)] of the B subunit of Shiga toxin, was inserted into the lamB gene of Escherichia coli and expressed in the S. flexneri vaccine strain. The chimeric LamB protein functioned normally and the epitope was expressed at the surface of the bacteria. The animals immunized with the live bacteria, expressing the epitope or sonicated lysates, showed a humoral response that was specific to the peptide (13-26 aa) and to the whole B subunit molecule. The elicited antisera neutralized the toxin activity on HeLa cells up to 40%, while the purified IgG fractions from the sera gave 90% neutralization.


Ultrastructural study on the adherence of Branhamella catarrhalis to oropharyngeal epithelial cell.

In the present study, it was observed that Branhamella catarrhalis adhere to the microplicae of the oropharyngeal epithelial cells. Both long and short microplicae patterns are present on the surface of oropharyngeal epithelial cells and the adherence ability of fimbriated Branhamella catarrhalis also varies according to the microplicae pattern. It was found that Branhamella catarrhalis attached more to one surface of the epithelial cell than to the other, suggesting that the presence of receptors are more on one surface than on the other. Branhamella catarrhalis did not attach to the mucus layer but directly to the epithelial cell surface. Ruthenium red staining specimen showed that Branhamella catarrhalis attached to a granular ruthenium red positive layer on the microplicae and also to a ruthenium red positive component, external to the unit membrane of the epithelial cell membrane.


Serological cross-reaction between Yersinia enterocolitica O9 and non-O1 Vibrio cholerae bio-serogroup Hakata and antigenic analysis of their relationship by their lipopolysaccharides.

Cross-agglutination and cross-agglutinin absorption experiments were carried out on non-O1 Vibrio cholerae bio-serogroup Hakata (Hakata) and Yersinia enterocolitica O9 (O9). It was shown that the O-antigen of Hakata was closely related to that of O9 in an a, b-a, c type of relationship. The antigenic relationship between the O-antigens of the two bacteria was analyzed by passive hemolysis (PH) and passive hemolysis inhibition (PHI) tests by using their lipopolysaccharides (LPS) as antigen for sensitizing sheep red blood cells (SRBC) and, in the case of the latter, as an inhibitor in a PH system consisting of LPS-coated SRBC, guinea-pig complement and anti-Hakata or O9 antiserum, both unabsorbed and absorbed with the heterologous Hakata or O9 antigen. In the PH experiment, unabsorbed anti-Hakata antiserum had hemolytic titers of 126,100 and 2,600 against Hakata- and O9-LPS-coated SRBC, respectively, and anti-O9 antiserum had hemolytic titers of 19,400 and 38,800, respectively, against these SRBC. The PH experiment showed that anti-O9 antiserum contains a hemolysin reacting with the heterologous Hakata antigen at a high titer (19,400), while anti-Hakata antiserum contains a hemolysin reacting with the heterologous O9 antigen at a significant titer (2,600). The former was completely removed from anti-O9 antiserum with the Hakata antigen and the latter from anti-Hakata antiserum with the O9 antigen. Thus, serological cross-reactivity was demonstrated between the Hakata and O9 strains.


Prevalence of hepatitis C virus infections in dialysis patients and their contacts using a second generation enzymed-linked immunosorbent assay.

The prevalence of antibodies to the hepatitis C virus (HCV) was determined in 498 hemodialysis patients from three german dialysis units, 121 staff members and 42 family members using an enzyme-linked immunosorbent assay (ELISA) of the second generation which detects antibodies to a structural (C22) and to non-structural (C33c, C100, 5-1-1) recombinant antigens to HCV. Using the second generation ELISA 115 patients (23.1%) were anti-HCV positive versus 77 (15.5%) when sera were tested by an ELISA of the first generation containing only a non-structural antigen (C100). In 34 of these 40 discordant sera antibodies against at least one viral protein was found by a recombinant immunoblot assay. Of 5 sera containing antibodies to only one viral protein (C22) 3 were HCV RNA positive by polymerase chain reaction. Epidemiological evaluation of the patients revealed that the prevalence of anti-HCV was correlated to the duration of dialysis but not to the number of blood transfusions. Of 121 staff members 2 (1.6%) and 2 of 42 family members (4.7%) were positive indicating a low risk of the patients' contacts of acquiring HCV infection.


Neural subsystems for object knowledge.

Critical issues in the cognitive neuroscience of language are whether there are multiple systems for the representation of meaning, perhaps organized by processing system (such as vision or language), and whether further subsystems are distinguishable within these larger ones. We describe here a patient (K.R.) with cerebral damage whose pattern of acquired deficits offers direct evidence for a major division between visually based and language-based higher-level representations, and for processing subsystems within language. K.R. could not name animals regardless of the type of presentation (auditory or visual), but had no difficulty naming other living things and objects. When asked to describe verbally the physical attributes of animals (for example, 'what colour is an elephant?'), she was strikingly impaired. Nevertheless, she could distinguish the correct physical attributes of animals when they were presented visually (she could distinguish animals that were correctly coloured from those that were not). Her knowledge of input stimulus. To explain this selective deficit, these data mandate the existence of two distinct representations of such properties in normal individuals, one visually based and one language-based. Furthermore, these data establish that knowledge of physical attributes is strictly segregated from knowledge of other properties in the language system.


ATP receptor-mediated synaptic currents in the central nervous system.

Until now, the only well documented, fast excitatory neurotransmitter in the brain has been glutamate. Although there is evidence for adenosine 5'-triphosphate (ATP) acting as a transmitter in the peripheral nervous system, suggestions for such a role in the central nervous system have so far not been supported by any direct evidence. Here we report the recording of evoked and miniature synaptic currents in the rat medial habenula. The fast rise time of the currents showed that they were mediated by a ligand-activated ion channel rather than a second messenger system, thus limiting the known transmitter candidates. Evidence was found for the presence on the cells of glutamate, gamma-aminobutyric acid, acetylcholine and ATP receptors, but not for 5-hydroxytryptamine (5HT3) or glycine receptors. The evoked currents were unaffected by blockers of glutamate, gamma-aminobutyric acid or acetylcholine receptors but were blocked by the ATP receptor-blocker, suramin and the desensitizing ATP receptor-agonist alpha,beta-methylene-ATP. Our evidence identifies for the first time synaptic currents in the brain, mediated directly by ATP receptors.


[The Aachen Aphasia Bedside Test--criteria for validity of psychologic tests].

The Aachen Aphasia Bedside Test (AABT) has been developed in order to examine aphasic patients within the first 4 to 6 weeks after onset of illness. The psychometric properties of the AABT were established by repeated examination of 82 acute stroke patients, ratings by 20 raters on the basis of 10 videotapes, repeated examination of 28 chronic aphasics three times with an interval of 2 days, and parallel examination of 47 chronic aphasic patients with the AABT and the Aachen Aphasia Test (AAT), administered on the same day. Objectivity, reliability and validity of the AABT were highly rated, indicating its usefulness in acute stroke cases. Data on the 82 acute stroke patients showed that an initial prognosis can be made as early as the fourth day after the stroke.


The neurotoxic effects of methamphetamine on 5-hydroxytryptamine and dopamine in brain: evidence for the protective effect of chlormethiazole.

Studies were undertaken in mice and rats on the neurotoxic effects of methamphetamine on dopaminergic and 5-hydroxytryptaminergic neurones in the brain and the neuroprotective action of chlormethiazole. In initial studies, mice were injected with methamphetamine (5 mg/kg, i.p.) at 2 hr intervals, to a total of 4 times. This procedure produced a 66% loss of striatal dopamine and a 50% loss of tyrosine hydroxylase activity 3 days later. Chlormethiazole (50 mg/kg, i.p.), given 15 min before each dose of methamphetamine, totally prevented the methamphetamine-induced loss of tyrosine hydroxylase activity and partly prevented the loss of dopamine. Phencyclidine (20 mg/kg, i.p.), given in place of chlormethiazole, also prevented the loss of tyrosine hydroxylase. Administration to rats of 4 doses of methamphetamine (15 mg/kg, i.p.) at 3 hr intervals resulted in a 75% loss of striatal dopamine 3 days later and a similar loss of 5-HT and 5-HIAA in cortex and hippocampus. Chlormethiazole (50 mg/kg, i.p.), given 15 min before each injection of methamphetamine, protected against the loss of dopamine and indoleamine content, in the respective regions. Pentobarbital (25 mg/kg, i.p.) also provided substantial protection but diazepam (2.5 mg/kg, i.p.) was without effect. Confirming earlier studies, dizocilpine (1 mg/kg) also provided substantial protection against the methamphetamine-induced neurotoxicity. Preliminary data indicated that chlormethiazole was not neuroprotective because of a hypothermic action. These data therefore demonstrate that chlormethiazole is an effective neuroprotective agent against methamphetamine-induced neurotoxicity and extend the evidence for the possible value of this drug in preventing neurodegeneration.


Effects of short- and long-term administration of fluoxetine on the monoamine content of rat brain.

The effects of repeated doses of fluoxetine over time and dose-responses of the content of indoles and catecholamines and metabolism, were examined in rats in relation to the concentrations of the parent compound and its active metabolite norfluoxetine in brain. Brains were removed for assays of the regional content of monoamines and concentrations of drugs 24 hr after the last dose on days 1, 7 and 21 of a twice-daily schedule of fluoxetine (15 mg/kg, i.p.). Measurements were also taken 1 week after the last dose (7.5 and 15 mg/kg, b.i.d.) of the 21-day regimen. On day 1 fluoxetine did not change the content of serotonin (5-HT) but reduced the concentrations of 5-hydroxyindolacetic acid (5-HIAA) in the hippocampus and cortex, compatible with the action of a blocker of the uptake of 5-HT. Continued injections of fluoxetine, however, significantly reduced 5-HT in the brain of the rat, the depletion being significant on days 7 and 21 in the hippocampus and cortex, respectively. The content of indoles remained significantly decreased for at least a week after the last dose of fluoxetine in the 21-day regimen, although the concentrations of 5-HIAA (but not 5-HT) totally recovered at the smaller dose (7.5 mg/kg) in all regions of the brain (cortex, hippocampus and striatum). In spite of slight changes in the concentrations and metabolism of dopamine (DA) in the striatum, 24 hr after the last dose (15 mg/kg), treatment with drug had no significant long-term effects on the content of catecholamines in these regions of the brain.(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of activated platelets using activation-dependent monoclonal antibody (SZ-51) in clinical disorders.

Platelets may become activated in a number of clinical disorders and participate in thrombus formation. A direct test for activated platelets in whole blood has been developed by radioimmunoassay with 125I labelled SZ-51, an antibody specific for an alpha-granule membrane protein (GMP-140) that associates with the platelet surface during secretion. The assay with sufficient sensitivity can be applied satisfactorily to detect as few as 2% activated platelets. In 50 normal subjects, minimal GMP-140 molecules per platelet were expressed on the surface of circulating platelets. The expression of GMP-140 molecules was increased transiently in 10 patients undergoing cardiopulmonary bypass, especially at the end of bypass. Evaluation of 18 patients with epidemic hemorrhagic fever (EHF) showed that the number of GMP-140 molecules on the platelet surface increased significantly, particularly at the shock phase, and closely related to the four different phases of EHF. In 9 patients suffered from acute myocardial infarction (AMI), the number of GMP-140 molecules changed with the procession of AMI and the highest occurred at the 48th h after AMI. The GMP-140 molecules were also increased in patients with asthma attack (n = 14), but not in patients with idiopathic thrombocytopenic purpura (n = 11). Considered together, these results suggest that activated platelets can be measured reliably in whole blood using radiolabeled SZ-51 antibody and the detection of activated platelets is potentially useful in identifying patients with certain thrombotic disorders.


Immunological characterization of the region of tau protein that is bound to Alzheimer paired helical filaments.

Tau protein is known to be present in the paired helical filaments (PHFs) of Alzheimer brains. This study investigated the fragments of tau protein that remain bound to pronase-treated PHFs and conditions that lead to the release of these tau fragments from the core structure of the PHF. Antibody 423 reacted with PHFs and with fetal rat tau but not with adult rat tau, pig tau, or recombinant human tau. Three other antibodies that react with the tubulin binding region of tau only reacted with PHFs after they were disrupted with formic acid or guanidine. Other antibodies that recognize tau sequences C terminal to the tubulin binding region also recognized pronase-treated PHFs. Antibodies SMI34 and T3P that recognize phosphorylated epitopes were reactive with pronase-treated PHFs. Tau fragments from the PHF were solubilized by acid or guanidine treatment. These findings suggest that the fragments of tau that are bound to PHFs and protected from pronase digestion include sequences from the tubulin binding region to the C terminus of tau. In addition, some of these sequences appear to be conformationally or post-translationally modified.


Platelets and vitronectin: immunocytochemical localization and platelet interaction with exogenously added vitronectin.

Vitronectin stores were localized in human platelets by immunofluorescence, confocal laser microscopy, and immunoelectron microscopy. In ultrathin cryosections, the immunogold labelling was mainly observed in the alpha-granules, although occasional immunolabelling of the platelet surface was present. The interaction between thrombin-stimulated platelets and exogenously added vitronectin was also explored. This vitronectin had been purified by heparin-affinity chromatography. Binding of vitronectin to thrombin-stimulated platelets was demonstrated by flow cytometry. The immunoelectron microscopical studies revealed that this binding was mainly restricted to clumps of released alpha-granular material on the platelet surface. The possible significance of the interaction between exogenous vitronectin and released platelet proteins is discussed.


Chronic dietary pergolide preserves nigrostriatal neuronal integrity in aged-Fischer-344 rats.

Pergolide, a potent D2 presynaptic agonist with postsynaptic D2 agonist activity and some D1 agonist activity was administered in the diet (0.5 mg/kg/day) of male Fischer 344 rats from age 3 to age 26 months. We hypothesized that the potent D2 presynaptic activity would reduce the baseline release of dopamine (DA) and thereby slow the formation of toxic oxidative metabolites that lead to age-related deterioration of nigrostriatal DA neurons. Pair-fed rats served as controls. We observed age-related losses of fluorescent DA cell bodies in the substantia nigra pars compacta and of fluorescent DA terminals in the striatum; chronic pergolide administration prevented these losses. Pergolide administration also prevented the age-related diminution of DA fluorescence intensity in substantia nigra cell bodies. A large decline in 3H-DA uptake with age was partially prevented by pergolide administration. We found no age-related alteration in the concentration of DA in the striatum and pergolide did not alter this concentration. Pergolide treatment resulted in only minor alterations in striatal 3H-spiperone binding and no change in dendritic arborizations of either DA substantia nigra neurons or medium spiny striatal neurons. Pergolide administration also prevented an age-related decline in circulating FSH levels. The uptake data and quantitative morphological findings suggest that pergolide administration in the diet for 2 years exerts a protective effect on age-related deterioration of DA nigrostriatal neurons. This finding was consistent with clinical reports of a subset of patients with Parkinson's disease in whom long-term efficacy of pergolide therapy is observed.


[Early results of the treatment of Hodgkin's disease in adults using the seven-drug cytostatic protocol].

65 adult patients with Hodgkin's disease were treated acc. to multidrug protocol proposed by I. Koza et al. including doxorubicin, vincristine, vinblastine, bleomycin, procarbazine, lomustine and prednisone. 4 drugs (3 cytostatics and prednisone) including cycles were given repeatedly every 4 weeks. In the group of first line therapy (39 persons) 61.5% CR and 23% PR was obtained i.e. 84.5% therapeutic responses. The protocol used as salvage therapy resulted in 30% CR and 23% PR. Undesirable gastrointestinal effects were observed less commonly than after ABVD and CVPP schemes but myelosuppressive effect was more often seen and required attenuation of cytostatic drugs doses in the further cycles of therapy. The considered protocol seems to be a valuable one approaching to -but not achieving- the results of MOPP and ABVD schemes: is better tolerated because of elimination of strong emetic cytostatics (chlormethine and decarbazine) but late toxicity could be evaluable only after several years.


[Long-term mexiletine treatment of ventricular arrhythmia in patients after myocardial infarction].

Eighty two patients with ventricular extrasystole (VES) after myocardial infarction had been treated with antiarrhythmic drugs in the half (41 patients) with mexiletine (M) and in the half (41 patients) with other ones. The mortality rate in the M-group was twice time lower than in the other, however this difference was statistically not significant. M was mostly effective in VES of III and IVb degree after Lown's classification and did not cause any deterioration of the left ventricle function.


[Effect of epirubicin on the heart conduction system in patients with Hodgkin's disease].

24 patients (12 women and 12 men) aged 17 to 70 years with Hodgkin's disease in the clinical stages IIA-IVB were treated according to the polychemotherapeutic schemes containing epirubicin (epiDX). Using the 24-hour ambulatory ECC on the day before and the day of drug application as well as after completion of the therapy, heart rate, arrhythmias and conduction abnormalities were estimated. There was statistically insignificant decrease in heart rate (mean, minimal and maximal) on the day of epiDX administration and after therapy completion. Arrhythmias in form of supraventricular and ventricular extrasystoles including bigeminy and couplets of VES in one patient and two episodes of atrial tachycardia in another one on the day of drug injection were noticed. In 2 young women occurred interruption of sinus rhythm lasting longer than 3 sec., due to sinus arrest in one and due to 2:1 sinus exit block in the second one. The above mentioned disturbances were in majority of patients not associated with clinical symptoms. The authors conclude that the reduction in heart rate and SVES formation might be in the majority of patients neurogenic but in some of them as well as VES generation result from epiDX action.


Nurses' perceptions of the dimensions of nursing care episodes.

The purpose of this exploratory study was to describe the dimensions of 17 nursing care episodes as perceived by nurses. The episodes represented recurrent nursing care situations in the postoperative period. Nineteen nurses judged the dissimilarity of all possible pairs of the episodes. An additional 14 nurses rated each episode on 10 attribute scales. Multidimensional scaling (MDS), was used to analyze nurses' dissimilarity judgments. A three-dimensional MDS configuration, which accounted for 51% of the variance of nurses' optimally scaled data, was chosen to describe the structure of the episodes. To interpret the dimensions of the MDS configuration objectively, multiple regression was used to correlate the attributes, one at a time, with the coordinates of the episodes in the configuration. The dimensions of the MDS configuration were interpreted as (a) the degree of independence patients could achieve in the episodes, (b) the nursing knowledge or skill needed in the episodes, and (c) the degree to which nurses could individualize the episodes for patients. Information obtained from the MDS analysis can be used to help nurses (a) explicitly evaluate and communicate their expectations for patients regarding activities to promote independence and (b) better articulate the specialized knowledge and skills needed to provide nursing care.


Masculinization of growth hormone (GH) secretory pattern by dihydrotestosterone is associated with augmentation of hypothalamic somatostatin and GH-releasing hormone mRNA levels in ovariectomized adult rats.

The role of androgen in the sexual dimorphism in hypothalamic growth hormone (GH)-releasing hormone (GHRH) and somatostatin (SS) gene expression was examined in rats. In the first study, the SS and GHRH mRNA levels were measured in both male and female rats at 4, 6, 8, and 10 weeks of age. A significant sex-related difference in the SS and GHRH mRNA levels was observed after 8 weeks of age, when sexual maturation is fully attained. Male rats had higher SS and GHRH mRNA levels than the female rats. In the second study, adult ovariectomized rats received daily injection of dihydrotestosterone (DHT), nonaromatizable testosterone, at a dose of 2 mg/rat for 21 days. The DHT treatment masculinized the GH secretory pattern, which was indistinguishable from that of intact male rats, and simultaneously augmented the SS and GHRH mRNA levels. The DHT treatment of ovariectomized rats after hypophysectomy significantly raised the level of SS mRNA, but not that of GHRH mRNA compared to the control animals. These findings suggest that the activation of the SS gene expression through androgen receptor plays an important role in the maintenance of sexual dimorphism in GH secretion in rats.


Peptide immunoreactive neurons in the amygdala and the bed nucleus of the stria terminalis project to the midbrain central gray in the rat.

The central nucleus of the amygdala, bed nucleus of the stria terminalis, and central gray are important components of the neural circuitry responsible for autonomic and behavioral responses to threatening or stressful stimuli. Neurons of the amygdala and bed nucleus of the stria terminalis that project to the midbrain central gray were tested for the presence of peptide immunoreactivity. To accomplish this aim, a combined immunohistochemical and retrograde tracing technique was used. Maximal retrograde labeling was observed in the amygdala and bed nucleus of the stria terminalis after injections of retrograde tracer into the caudal ventrolateral midbrain central gray. The majority of the retrogradely labeled neurons in the amygdala were located in the medial central nucleus, although many neurons were also observed in the lateral subdivision of the central nucleus. Most of the retrogradely labeled neurons in the BST were located in the ventral and posterior lateral subdivisions, although cells were also observed in most other subdivisions. Retrogradely labeled neurotensin, corticotropin releasing factor (CRF), and somatostatin neurons were mainly observed in the lateral central nucleus and the dorsal lateral BST. Retrogradely labeled substance P-immunoreactive cells were found in the medial central nucleus and the posterior and ventral lateral BST. Enkephalin-immunoreactive retrogradely labeled cells were not observed in the amygdala or bed nucleus of the stria terminalis. A few cells in the hypothalamus (paraventricular and lateral hypothalamic nuclei) that project to the central gray also contained CRF and neurotensin immunoreactivity. The results suggest the amygdala and the bed nucleus of the stria terminalis are a major forebrain source of CRF, neurotensin, somatostatin, and substance P terminals in the midbrain central gray.


Effects of an interferon inducer bropirimine on bleomycin-induced lung fibrosis in hamsters.

The antifibrotic effect of an interferon inducer, bropirimine (2-amino-5-bromo-6-phenyl-4(3H)-pyrimidinone, ABPP) was evaluated in bleomycin (BLM)-hamster model of lung fibrosis. ABPP is an orally active biological response modifier and has immunomodulatory, antiviral, and antineoplastic activities. The hamsters were randomized in four groups and treated with either bropirimine (100 mg/kg, intraperitoneally) suspended in carboxymethylcellulose (CMC, 10 mg/10 mg/ml) or CMC alone each day for sixteen days. After two days, the hamsters received either single bolus of BLM (7.5 U/5 ml/kg) or an equivalent volume of saline by the transoral endotracheal route. Groupings were assigned as: CMC + saline (CS), ABPP + saline (AS), CMC + BLM (CB) and ABPP + BLM (AB). Animals were sacrificed at fourteen days after intratracheal installation of either BLM or saline. Their lungs were lavaged and processed for morphometric and biochemical studies. ABPP had little effect in preventing BLM-induced weight loss and lung injury. ABPP was found to reduce the BLM-induced accumulation of collagen in the lung as measured by hydroxyproline content. The hamsters in AB group had significantly less collagen than the hamsters in CB group: 995 and 1157 micrograms hydroxyproline/lung, respectively. Administration of ABPP prevented the BLM-induced increase in the lung prolyl hydroxylase activity. The total number of monocytes and eosinophils recovered from the bronchoalveolar lavage fluid (BALF) of the AB group were significantly lower than that of the animals in CB group. However, the BALF supernatant protein content from animals in AB group (7.9 mg/lung) was significantly higher than that of CB group (4.5 mg/lung).(ABSTRACT TRUNCATED AT 250 WORDS)


Rod opsin cDNA sequence from the sand goby (Pomatoschistus minutus) compared with those of other vertebrates.

The absorbance spectra of rods from the sand goby were measured by using microspectrophotometry. Analysis of the averaged spectra shows that the rod visual pigment has a maximum absorbance (lambda max) at approximately 501 nm. A sand goby retinal cDNA library was constructed and then screened with a partial sand goby rod opsin clone obtained by the polymerase chain reaction (PCR). The screening of the library yielded a full length rod opsin clone. The cDNA sequence and deduced amino acid sequence of this clone are compared with those of other vertebrate rod opsins.


Galanin inhibits rat pancreatic amylase release via cholinergic suppression.

The effects of galanin on pancreatic exocrine function were examined using rat pancreatic tissues. In anesthetized rats, galanin (40 micrograms/kg/h) decreased amylase secretion stimulated by 2-deoxy glucose (5.8 +/- 0.1 vs. 3.1 +/- 0.1 times basal) and cholecystokinin octapeptide (21.5 +/- 0.6 vs. 16.8 +/- 0.5), while not inhibiting bethanechol-stimulated secretion. In dispersed acini, there was no effect of galanin alone (10(-8) to 10(-13) M) on amylase release, nor did galanin (10(-6) or 10(-8) M) coincubation affect amylase release stimulated by bethanechol (10(-3) to 10(-7) M) or CCK-8 (10(-8) to 10(-13) M). Using pancreatic lobules, coincubation with galanin (10(-6) M) suppressed 75 mM KCl-stimulated amylase secretion and ACh release (10.1 +/- 0.6% vs. 7.3 +/- 0.4%). Veratridine-stimulated (10(-4) M) amylase secretion and ACh release (12.4 +/- 1.7% vs. 8.5 +/- 0.7%) were similarly diminished.


Cloning of ShIII (Shaw-like) cDNAs encoding a novel high-voltage-activating, TEA-sensitive, type-A K+ channel.

Transient voltage-dependent potassium (K+) currents, also known as A currents, have been of great interest to neurophysiologists due to their special roles in neuronal excitability. Several cDNAs encoding transcripts expressing A currents have been characterized. Recently, a cDNA (KShIIIC or Raw3) was isolated which expresses an unusual A current that is highly sensitive to TEA, and activates at potentials more positive than -20 mV. Channels containing this protein may have specialized roles in modulating the electrical behaviour of neurons. Here we report the isolation and characterization of two rat cDNAs corresponding to two alternatively spliced transcripts (KShIIID.1 and KShIIID.2) from another gene (KShIIID) of the same subfamily as KShIIIC, the ShIII or Shaw-related gene subfamily. KShIIID.1 also expresses an unusual high-voltage-activating, TEA-sensitive A-type channel. There are, however, significant differences between KShIIIC and KShIIID channels which may have interesting functional consequences. The two most important differences are: (i) KShIIID channels conduct in the steady-state over a much broader window of potentials than KShIIIC; this reflects differences between the kinetic schemes of the two channels; and (ii) KShIIID inactivates with significantly slower kinetics than KShIIIC. The identification of KShIIID transcripts contributes to our knowledge of the molecular components that may determine the functional diversity of A currents and provides exciting opportunities to increase our understanding of the structure and function of K+ channels.


Distribution and effects of neuropeptide Y, vasoactive intestinal peptide, substance P, and calcitonin gene-related peptide in human middle meningeal arteries: comparison with cerebral and temporal arteries.

A sparse to moderate supply of nerve fibers containing neuropeptide Y-like immunoreactivity (NPY-LI), vasoactive intestinal polypeptide (VIP-LI), substance P (SP-LI), and calcitonin gene-related peptide (CGRP-LI) was demonstrated in the walls of human middle meningeal arteries. Comparison with similar studies on human cerebral and temporal arteries indicated a similar distribution and density. The immunoreactive material in all three arterial regions was characterized by reversed-phase high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA). The major peak of NPY-LI, VIP-LI, SP-LI, and CGRP-LI in each extract eluted approximately with the same elution volume as that of the corresponding synthetic analogues. The concentration of NPY in the middle meningeal arteries was lower as compared to the temporal arteries. Low concentrations of SP-LI and CGRP-LI were found in the middle meningeal arteries as compared to the cerebral arteries. In isolated ring segments of human middle meningeal and cerebral arteries, NPY caused vasoconstriction but did not potentiate the contractile response of noradrenaline. In the temporal artery, NPY did not induce contraction but potentiated the vasoconstrictor response to noradrenaline. Vasoactive intestinal polypeptide, peptide histidine methionine-27, SP, neurokinin A, and CGRP relaxed all three types of cephalic arteries. The peptide effects were not antagonized by propranolol, atropine, or cimetidine. Comparison of the responses to VIP and SP of vessels from the different regions showed a similar pattern of reactivity. The response to SP was slightly (p less than 0.05) more potent, whereas the responses to CGRP were less potent in the middle meningeal as compared to that in cerebral (p less than 0.005) vessels.


Protection by a new synthetic protease inhibitor, ONO3307, of the rat exocrine pancreas during acute edematous pancreatitis induced by a supramaximal dose of caerulein in comparison with FOY007.

The present study investigated the protective effects of the new potent synthetic protease inhibitors, ONO3307 (4-sulfamoyl phenyl-4-guanidinobenzoate methanesulfonate) and FOY007 (gabexate misilate), on the exocrine pancreas in rat caerulein-induced acute pancreatitis in both in vitro and in vivo experiments. These protease inhibitors prevented hyperamylasemia, pancreatic edema, congestion of amylase, redistribution of lysosomal enzyme in acinar cells, and lactic dehydrogenase (LDH) discharge from dispersed acini. They also inhibited the cathepsin B leakage from the lysosomes in a dose-dependent manner in doses of 2-10 mg/kg.h of ONO3307 and 20-50 mg/kg.h of FOY007. These results indicate that both ONO3307 and FOY007 exert protective effects against pancreatitis at subcellular levels in lysosomes and cellular or organelle membranes. Proteases appear to be important in the pathogenesis and development of acute pancreatitis, and low-molecular-weight protease inhibitors may be of clinical use in the treatment of acute pancreatitis.


Voltage-dependent calcium current and the effects of adrenergic modulation in rat aortic smooth muscle cells.

Smooth muscle cells from rat aorta were cultured in defined, serum-free medium and studied using whole-cell patch-clamp techniques. Under conditions designed to isolate currents through Ca channels, step depolarizations produced inward currents which were fast in onset and inactivated rapidly, with little sustained inward current being observed. Both Ni and Cd blocked these currents, with Ni being effective at 50 microM. Removal of external Na or addition of 1 microM tetrodotoxin had no effect. Peak inward currents were attained at about -15 mV, with half-maximal activation at -41 mV using -80 mV holding potentials. The transient inward currents were reduced by depolarized holding potentials, with half-maximal steady-state inactivation at -48 mV. In three of the 98 cells studied, small maintained inward currents were observed with a -40 mV holding potential. The Ca channel antagonist nicardipine (5 microM) blocked the transient inward current while neither of the dihydropyridine Ca channel agonists S(+)202 791 and (-)BAY K 8644 produced a significant augmentation of sustained inward current. At 10 microM, both noradrenaline and adrenaline but not phenylephrine decreased the peak inward current. This inhibition was unaffected by a variety of adrenoceptor antagonists and was also observed when internal solutions having high Ca buffering capacity were used, but was absent when GDP-beta-S instead of GTP was included in the pipette solution. The main conclusions from this study are that under our cell culture conditions, rat aortic smooth muscle cells possess predominantly a transient, low-threshold-activated inward Ca current and that this Ca current is inhibited by certain adrenoceptor agonists but with a quite atypical adrenoceptor antagonist pharmacology.


Housing influences exploration and social interaction of control and DSP-4-treated rats.

Exploratory behavior and social interaction were investigated in rats that were reared in different social environments following neonatal injection with either water vehicle or the norepinephrine neurotoxin, DSP-4. At weaning, they were placed in a familiar or novel bedding type and were housed in either vehicle control-only, DSP-4-only, or mixed vehicle control and DSP-4 groups for 10 days. They were then observed in three different situations: the home cage, the cage of an unfamiliar rat, and an open field. Compared to rats housed in vehicle control-only or DSP-4-only groups, rats housed in mixed DSP-4 and vehicle control groups showed elevated exploration behavior in the home cage. Also, rats housed in mixed groups in the familiar bedding, but not the novel one, showed abnormally low levels of rearing in an open field test and reduced social interaction with unfamiliar rats. The implications of these results for a new animal model of anxiety are discussed.


Incidental finding of rectal carcinoma during transrectal US for prostatic diseases.

During transrectal ultrasound (TRUS), rectal carcinoma was an incidental finding in seven patients among a series of 5,000 TRUS examinations. TRUS was performed in seven patients with symptoms characteristic of prostatic diseases. All seven patients underwent examination by at least one physician before TRUS and, except for abnormal prostatic findings, no tumors were detected during digital rectal examination (DRE). The tumors were clearly visualized with TRUS and were easily palpated during DREs performed after TRUS. They were large and were located mainly along the posterior and lateral walls of the rectum. All the tumors were diagnosed by means of proctoscopy; the biopsy findings were positive, and the pathologic staging indicated advanced disease: adenocarcinoma of the rectum with a minimum grade of Dukes C. It is recommended that, in addition to evaluation of scans obtained in the transverse plane, the multiplane transducer be used to evaluate the longitudinal plane of the rectum for detection of possible undiscovered tumors.


Characterization of porcine gastric galanin.

The presence of a peptide capable of producing powerful contractions of rat small intestinal smooth muscle was detected in chromatographic fractions derived from porcine gastric corpus extracts. The pharmacological characteristics of this entity suggested that it might be galanin and on its purification to homogeneity, amino acid composition and sequence analysis demonstrated the identify of the gastric and intestinal forms of galanin. The presence of galanin in the gastric corpus tissue and its ability to affect gastric smooth muscle activity, gastrin release, and gastric acid secretion suggest potential important physiological roles for galanin in the stomach.


Morphological evidence for a substance P projection from medial septum to hippocampus.

The medial septal nucleus provides one of the major afferents to the hippocampal formation. The two major types of neurons present in the medial septum are cholinergic and GABAergic, but other types of neurons are also present. A small population of substance P-containing neurons is present along the border between the medial and lateral septum, but it is unclear whether these project to the hippocampus. The present study, by employing both anterograde and retrograde tracing techniques, combined with immunocytochemistry for substance P, provides direct morphological evidence for a substance P projection from the lateral region of medial septum to a portion of CA2/3a, which is restricted to the mid-septotemporal portion of the hippocampus.


Dexfenfluramine treatment and hypothalamic neuropeptides in diet-induced obesity in rats.

Neuropeptide Y (NPY) is a powerful appetite stimulant, and hypothalamic concentrations rise after food deprivation and in experimental diabetes. Serotonergic drugs such as dexfenfluramine are inhibitors of feeding. We measured hyothalamic NPY and NPY mRNA, along with galanin, neurotensin, and somatostatin in chow-fed rats and in rats with dietary obesity, and examined the effect of dexfenfluramine on these peptides in this model. Sixty-five rats were fed a palatable diet (condensed milk, sucrose and chow) for 6 weeks, which produced significant weight gain compared to twenty fed standard chow (145.1 +/- 2.3 g vs. 113.4 +/- 3.2 g, p less than 0.001). Groups of animals were treated for 7 days or 28 days with dexfenfluramine (1.8 mg/kg/day) or saline intraperitoneally via miniosmotic pumps. Hypothalami were dissected into medial and lateral blocks, and NPY, galanin, neurotensin, and somatostatin were measured by radioimmunoassay. Neuropeptide Y mRNA was measured by Northern blotting. Hypothalamic NPY was significantly higher in the palatable diet group compared to chow-fed controls (medial hypothalamus: 86.6 +/- 7.6 vs. 65.7 +/- 4.0 pmol/g tissue, p less than 0.02, lateral hypothalamus 71.2 +/- 6.6 vs. 53.1 +/- 3.6 pmol/g tissue, p less than 0.05), but NPY mRNA was unchanged. Although dexfenfluramine was effective at reducing weight gain in the animals fed the palatable diet, this did not result in any changes in the hypothalamic neuropeptides measured. Neuropeptide Y may be of importance in diet-induced obesity but the weight loss produced by dexfenfluramine in such animals is not mediated by changes in hypothalamic NPY.


Amplification of dengue 2 virus ribonucleic acid sequence using the polymerase chain reaction.

The polymerase chain reaction (PCR) has been adapted to the amplification of dengue type 2 virus (DEN2) nucleic acid sequences. A pair of 20-mer oligonucleotides were designed and synthesized based on conserved sequence blocks of DEN2 strains isolated from different geographical areas. RNA samples were prepared from two DEN2 strains, prototype New Guinea C (NGC) and local isolate Hainan 98 (HN98). The reverse transcription step was performed for cDNA synthesis before the standard PCR procedures. The amplified products were fragments about 476 bp in length, corresponding to the upper one third of DEN2 envelope gene (E1 to E476 nt). Specificity of the amplification products was confirmed by "nested" PCR using the internal primers and by Southern and dot blot hybridization to cloned DEN2 cDNA probes following agarose gel electrophoresis. Further improvement and the potential application of the methods in study of dengue virus RNA are discussed.


Serological and virological studies of Japanese encephalitis in the Terai region of Nepal.

In 1987 and 1990, serum samples were collected from people living in the two districts (Itahari and Chitwan) of the Terai region of Nepal. Antibodies against Japanese encephalitis (JE) virus in these sera were detected by the hemagglutination inhibition (HI) and neutralization (N) tests. By the HI test, 26 out of 172 (15.1%) sera from Chitwan and 15 out of 137 (10.9%) sera from Itahari showed positive titers. Higher positive rates were shown by the N test, where 46 out of 172 (26.7%) sera from Chitwan and 22 out of 137 (16.1%) sera from Itahari had antibodies against JE virus. A JE strain was isolated from a blood specimen of a pig raised in Kathmandu. When the nucleotide sequence of the pre-M region of the strain was compared to the same region of the other JE virus strains reported, the highest similarity was observed to the strains isolated in Nepal in 1985. These results suggest that the Terai region has been an epidemic area of JE.


Studies on common antigenicities between the Belo Horizonte strain, Brazil of Schistosoma mansoni eggs and Biomphalaria snails by immunoelectrophoresis.

Immunoelectrophoretic studies of common antigenicities were carried out by using rabbit sera immunized against the Belo Horizonte strain of Schistosoma mansoni eggs and crude antigens of Biomphalaria snails and vice versa. With regard to common antigenicities between S. mansoni eggs and Biomphalaria snails, S. mansoni eggs produced 4 to 5 bands with Biomphalaria glabrata pigmentado, 3 to 4 bands with B. glabrata albino and only 1 band with B. straminea. In our laboratory, the infection rate of S. mansoni miracidia to B. glabrata pigmentado was 64.3% and 55.0% for B. glabrata albino, but B. straminea was not found to be susceptible to S. mansoni miracidia. It was observe that more bands were seen between S. mansoni egg and suitable hosts. Biomphalaria snail crude antigens were fractionated by Sephadex G-100 column, and each fraction antigen was tested with anti-S. mansoni egg sera by immunoelectrophoresis. As results, three fractions were collected form each snail strains. The common antigenicities between fractionated antigens from Biomphalaria snails crude antigens and anti-S. mansoni egg sera mostly existed in the first fraction and they were estimated to have molecular weights over 45,000.


[Epilepsy and thyrotoxicosis in a 4-year-old boy].

Hyperthyroidism in childhood has a relative incidence of 5%. The presence of epilepsy secondary to thyrotoxicosis is very unusual. We report the case of a four-year old boy with thyrotoxicosis due to Graves' disease. This patient developed a generalized tonic-clonic seizure followed by left sided partial motor status epilepticus. The EEG was markedly abnormal. The EEG was normal after five months of the ablative therapy. At the present time the patient is seizure-free without any antiepileptic medication and receiving replacement therapy with thyroxin due to post-ablation hypothyroidism. We conclude that this is the first reported case with this association in our country. We discuss the possible pathophysiological mechanism involved in the development of seizures in this patient.


[The role of serotonin, histamine and the kallikrein-kinin system in the pathogenesis of asphyxic attacks in bronchial asthma].

Altogether 300 patients suffering from bronchial asthma were examined for serotonin metabolism, the kallikrein-kinin system and histamine. A study was also made of the information relations of the indicated parameters to diverse groups of the signs using Shannon's technique. It has been revealed that serotonin and the kallikrein-kinin system are of importance in the pathogenesis of changes in the efficacy of the diffusion lung capacity in severe bronchial asthma. The authors discuss the significance of excessive accumulation of serotonin and bradykinin as a marker of the impairment of endothelial function of pulmonary vessels, specified by the damaging action of free radicals and immune complex allergic vasculitis as well. Histamine along with macrophages act as delimiters of allergic inflammation, with their action being mediated by negative feedback.


Heparin cofactor II deficiency in the elderly: comparison with antithrombin III.

We investigated heparin cofactor II (HC II) levels and their relationship to other haemostatic factors in the elderly in comparison with antithrombin III (AT III). We measured plasma HC II activity levels in 166 subjects aged from 61 to 99 years using a chromogenic method. HC II levels (94.4 +/- 18.5%) in the healthy elderly subjects were significantly (p less than 0.001) lower than in 40 healthy adult controls under 60 years of age (mean age: 51.5 years; 111.6 +/- 21.2%). HC II levels in the elderly subjects decreased further with age (r = 0.308, p less than 0.001) and the extent of the decrease was more marked than that for AT III (r = 0.179, p less than 0.05). There was no significant sex difference in HC II levels in the elderly. HC II levels correlated significantly with AT III levels and with acute phase reactants including sialic acid, fibrinogen, and PAI-1. HC II levels also correlated with factor VII, plasminogen, alpha 2-plasmin inhibitor, serum lipid, pseudocholinesterase, and albumin levels. These correlations were also found for AT III except active PAI-1 and tPA-PAI-1 complexes, but the correlations with acute phase reactants were stronger for HC II than AT III. We divided 154 elderly subjects into 4 groups by their pseudocholinesterase and albumin levels to estimate the effect of nutritional status on antithrombin activity in the elderly. HC II levels were normal in the elderly subjects with a good nutritional state (103 +/- 18%), but were significantly decreased in those with malnutrition (85 +/- 15%, p less than 0.001). AT III levels also showed the same tendency. These results indicate a decrease in the reserve capacity to inhibit thrombin generation at sites of atherosclerosis in response to trigger events. The deficiency of two major antithrombin factors in the elderly may indicate a tendency to thrombosis, especially in individuals with malnutrition. When considering the clinical significance of HC II, several other parameters, including age, nutritional status, hepatic synthetic ability, and the presence or absence of acute phase reaction should also be assessed.


Modulation of heparin cofactor II activity by glycosaminoglycans and adhesive glycoproteins.

Heparin cofactor II (HCII) is a specific thrombin inhibitor; its inhibitory activity is stimulated by heparin (Hep) and dermatan sulfate (DS). Vitronectin (VN), a heparin binding adhesive glycoprotein present in plasma and extracellular matrix, has been shown to decrease the stimulatory effect of Hep but not of DS on thrombin inhibition by HCII (Preissner and Sié, 1988). We analyzed the effect of glycosaminoglycans (GAGs) and GAG-binding proteins on the HCII/thrombin interaction in more detail. HCII was purified from the supernatant of barium citrate adsorbed normal human plasma by polyethylene glycol precipitation followed by affinity chromatography on heparin-Sepharose CL-6B and ion-exchange chromatography on a QAE-Sephadex A-50. Inhibition of thrombin by HCII was studied in the absence and presence of GAGs (hep 0.03-30 micrograms/ml, DS 0.05-50 micrograms/ml, heparan sulfate (HS) 0.05-50 micrograms/ml) in a chromogenic substrate assay using S-2366 as a thrombin substrate. The effects of VN and fibronectin (FN) on HCII stimulation by GAGs were determined. In addition to Hep and DS the inhibitory effect of HCII was stimulated by HS, however, to a lesser extent. Using 0.03U/ml thrombin and 1nM HCII the stimulatory effect of GAGs was completely inhibited when Hep (less than or equal to 0.3 micrograms/ml) was preincubated with VN (60 micrograms/ml) and decreased to less than 50% when HS (50 micrograms/ml) was preincubated with VN (60 micrograms/ml). VN had no effect on DS as already described previously.(ABSTRACT TRUNCATED AT 250 WORDS)


A longitudinal study on anthropometric and clinical development of Indian children adopted in Sweden. II. Growth, morbidity and development during two years after arrival in Sweden.

One hundred and fourteen consecutively recruited children adopted from India (60% girls) to Sweden were studied during 2 years after arrival, with examinations monthly the first 6 months and thereafter every 3 months. Sixty-two percent were below 1 year of age at arrival. There was a mean increase from -2.2 standard deviation scores (SDS) height/age to -0.7 SDS during the two years, and a similar development for weight/age, with no significant difference between boys and girls. The weight/height remained at around -0.8 throughout the study period. Those who had lowest height/age at arrival had the most marked catch-up, but remained smaller throughout the 2 years. The psychomotor development was initially delayed in nearly 30% of the children, mainly among those stunted and/or with very low weight at arrival. After 2 years the rate was at a level similar to Swedish children. In a sub-sample, birth weight was found to be correlated to subsequent height and weight development. Hepatitis B, salmonella, giardia lamblia, trichuris trichiura, ascaris and hymenolepis nana were still found in a small percentage after 2 years. Other morbidity was at the same level as in Swedish children. Adopted children who are stunted and/or have a very low weight at arrival should be followed up with special care, and infectious diseases found at arrival should be kept in mind for differential diagnosis at subsequent disease episodes.


Basal cell hyperplasia of prostate: an elusive lesion?

We report a case of basal cell hyperplasia of the prostate accompanying benign prostatic hypertrophy. The histogenesis of the basal cells as well as the histologic features and differential diagnosis of basal cell hyperplasia are reviewed.


[Quality control in the allergy laboratory--4 years "Allergy Ring Trial" in Austria].

In 1988 the Austrian Society for Allergology and Immunology initiated an external quality control program for the in vitro allergy diagnosis. In 12 mailings, 62 sera from allergic patients were sent to selected laboratories in order to determine total and antigen specific IgE according to the laboratory-specific methods. The values for total IgE varied considerably (73.9% were within +/- 1 SD, 94.3% within +/- 2 SD and 99.1% within +/- 2 SD, but only 0.9% beyond that). Sources of error were mainly attributable to inappropriate equipment and low quality reagents, but also bad test performance, with respect to personnel and the routines. In contrast, antigen specific IgE against pollen, mites, moulds, insect venoms, animal danders, drugs, parasites, environmental and food antigens revealed quite homogeneous results. Out of 1492 data, only 46 (3.1%) had to be declared as "wrong", and the variability of the RAST-classes was low. Whereas the quality of the reagents from all the different suppliers was not absolutely reliable at the beginning of the study, it improved considerably with time, as consequence of our complaints. The comparability of the methods for detecting total-IgE were non-satisfactory, whereas those for antigen specific IgE were generally good. The variety of methods employed (radio- and enzyme immunologic, fluoro- and nephelometric methods, etc.) should entail appropriate consequences, especially critical comparisons within one and the same laboratory; in addition, international standardization of the "normal" values should replace company-standards, and quality control programs for each test system before it is marketed should be mandatory. Our results confirm, that external control should be obligatory for any laboratory.(ABSTRACT TRUNCATED AT 250 WORDS)


[Familial Mediterranean fever. Case of a 48-year-old patient with recurrent abdominal pain].

A 48 year-old male patient from Turkey underwent laparotomy 13 years before admission to one unit because of persistent pain in the upper abdomen and fever. Subsequently, he was repeatedly admitted to surgical departments with recurrent upper abdominal pain and fever. The patient was admitted for medical investigation to our department with fever and left pleuritic pain. During this observation period he repeatedly had attacks of fever lasting for one day with leucocytosis. The diagnosis of familial Mediterranean fever was made. Therapy with colchicine (1.5 mg/day) led to complete remission, maintained over the follow-up period of 2 years to date.


[Various indicators of serotonin metabolism in the cerebrospinal fluid of patients with neurasthenia and cerebral arachnoiditis].

Thirty patients with cerebral arachnoiditis and 26 with neurasthenia were found to have differences in the content of serotonin, 5-hydroxyindoleacetic++ acid and melatonin in the cerebrospinal fluid, which depended in arachnoiditis on the degree of intracranial pressure elevation. The data on serotonin metabolism can be used in objective evaluation of the organic background in neurotic patients and serve for estimating the intensity of hypertension in patients with cerebral arachnoiditis.


Ultrastructural studies of the cells forming amyloid in the cortical vessel wall in Alzheimer's disease.

Ultrastructural studies of serial sections of the vessels with amyloid deposits in the brain cortex of patients with Alzheimer's disease showed that cells in the position of pericytes--perivascular cells--and perivascular microglial cells are producers of amyloid fibrils in the vascular wall. Three types of changes from normal are distinguishable in the vessel wall: (1) semicircular or circular thickening of vascular wall containing a large amount of amorphous material and various number of amyloid fibrils, (2) tuberous amyloid deposits containing both amorphous material and amyloid fibrils, some of the fibrils being arranged in strata and others arranged radially, and (3) amyloid star composed of a predominantly radial arrangement of bundles of amyloid fibrils and a less prominent amorphous component. A mixture of amorphous material and amyloid fibrils is present in cell membrane invaginations of perivascular cells, and occasionally perivascular microglial cells. Bundles of amyloid fibrils are found in altered cisternae of the endoplasmic reticulum and in the channels confluent with the infoldings of the plasma membrane of perivascular microglial cells. The amyloid deposition in the wall of the vessel causes degeneration of endothelial cells and the reduction of, and in some vessels obliteration of, the vessel lumen. In areas affected by amyloid angiopathy, extensive degeneration both of the neuropil and of neurons was observed. These changes were accompanied by astrogliosis. This study demonstrates similarities in amyloid formation in amyloid angiopathy and in beta-amyloid plaques in the neuropil and suggests that cells of the mononuclear phagocyte system of the brain (perivascular cells and perivascular microglia) are engaged in amyloid fibril formation.


Presenile dementia with progressive supranuclear palsy tangles and Pick bodies: an unusual degenerative disorder involving the cerebral cortex, cerebral nuclei, and brain stem nuclei.

Degeneration of heterogeneous systems in the central nervous system, with widespread distribution of argyrophilic neuronal fibrillary inclusions, was found in a patient with presenile dementia. Atrophy was circumscribed in the frontal and temporal lobes. Neuronal loss was severe in the basal ganglia, subthalamic nucleus, and substantia nigra. Immunocytochemical study using anti-phosphorylated tau and anti-ubiquitin antibodies in conjunction with ultrastructural observations revealed two types of inclusions: neurofibrillary tangles (NFTs) of progressive supranuclear palsy (PSP) in the Edinger-Westphal nucleus, locus coeruleus, cerebellar dentate nucleus, inferior olivary nucleus, and posterior horn of the spinal cord; and Pick bodies (PBs) in the atrophied cerebral cortex and red nucleus. PSP-type NFTs and PBs have been demonstrated in a single case for the first time. Despite their pathognomonic significance in certain disorders, we suggest that these inclusions may reflect a form of cytoskeletal disorganization, which is not entirely restricted to a single disease entity.


Modifications in myotendinous junction structure following denervation.

Changes in the structure of myotendinous junctions in response to peripheral nerve lesions are examined by transmission electron microscopy and morphometric analysis. Modifications in the folding of the plasma membrane at myotendinous junctions relative to the cross-sectional area of myofibrils terminating at the membrane are evaluated quantitatively using a morphometric analysis in which the muscle cell processes at the myotendinous junction are modeled as circular paraboloids. Denervated frog semitendinosus muscles were analyzed at 2, 4 and 8 weeks following denervation and compared to innervated, contralateral controls. No significant differences were found in relative folding of junctional plasma membranes between any two data sets, although myofibril diameter decreased over time following denervation. This shows that junctional plasma membrane and associated junctional structures, such as basement membrane, are removed from the myotendinous junction at a rate similar to that of myofibril thinning, thereby keeping constant the ratio between junction area and myofibril cross-sectional area. Electron microscopic observations indicate that 4 weeks post-denervation is the most active stage of junction remodeling of the time points sampled.


Infectivity of human T-lymphotropic virus type I to human nervous tissue cells in vitro.

Infectivity of human T-lymphotropic virus type I (HTLV-I) to human nervous tissue cells was explored using co-cultivation with X-irradiated, HTLV-I-producing MT2 cells. Examined cells included normal cerebellar cells, brain tumor cells (astrocytoma, medulloblastoma, meningioma, hemangioblastoma, and schwannoma), and various cell lines (astrocytoma, ependymoma, oligodendroglioma, medulloblastoma, and neuroblastoma). Successful HTLV-I infection was confirmed immunohistochemically using monoclonal antibodies to HTLV-I p19, p24, and pX product. All cell lines and primary cultures from normal cerebellar tissues and brain tumors could be infected with HTLV-I. Double immunostaining showed that glial fibrillary acidic protein-, S-100 protein- or vimentin-positive cells were susceptible to infection. Neurofilament- or neuron-specific enolase-positive cells in medulloblastoma could also be infected. Reverse-transcriptase assay revealed the productive infection in U251-MG (astrocytoma) and KG-IC (oligodendroglioma) lines. Co-cultivated U251-MG cells formed syncytial polykaryons after serial passages, and polymerase chain reaction assay detected HTLV-I genome in U251-MG syncytial polykaryons and p19+ mononuclear cells. HTLV-I viral RNA was also detected in infected U251-MG cells by in situ hybridization. These data show that HTLV-I may have a wide spectrum of infectivity in human nervous tissues.


Histochemical study of pituitary adenomas with Ki-67 and anti-DNA polymerase alpha monoclonal antibodies, bromodeoxyuridine labeling, and nucleolar organizer region counts.

The growth potential of 65 pituitary adenomas was determined by histochemical analysis with Ki-67 and anti-DNA polymerase alpha monoclonal antibodies, bromodeoxyuridine (BrdUdR) labeling, and counts of argyrophilic nucleolar organizer regions (Ag-NORs). The mean proliferating cell indices (PCIs) determined by Ki-67 and anti-DNA polymerase alpha and the BrdUdR labeling index (LI) were generally very low [1.0 +/- 0.2%, 1.1 +/- 0.2%, and 0.5 +/- 0.1% (+/- SE), respectively]. Apart from adrenocorticotropic hormone-positive adenomas, which had significantly higher indices, there were no statistically significant differences in the indices among the other subtypes of pituitary adenomas. Recurrent tumors had higher Ki-67 and DNA polymerase alpha PCIs and BrdUdR LIs (3.6%, 4.2%, 1.4%) than primary tumors (0.8%, 0.8%, 0.3%; P less than 0.005). The number of Ag-NORs did not correlate significantly with any of the three indices. The mean number of Ag-NORs was higher in nonfunctioning adenomas than in functioning adenomas (2.04 vs 1.66, P less than 0.005); among prolactin-positive adenomas, those treated preoperatively with bromocriptine had more Ag-NORs than untreated tumors (1.75 vs 1.57, P less than 0.005). These results suggest that the Ki-67 and DNA polymerase alpha PCIs and the BrdUdR LI predict the growth potential of individual pituitary adenomas, whereas the number of Ag-NORs appears to correlate with hormone production rather than with the proliferative potential.


Neuronal damage, glial response and cerebral metabolism after hypothermic forebrain ischemia in the rat.

We investigated the effect of 30 degrees C whole body hypothermia on neuronal injury, astroglial reactivity and intracellular pH in rats subjected to 15 min of forebrain ischemia. Experimental groups included: (1) normothermic ischemia (n = 8), ischemia induced under 37 degrees C body temperature, (2) hypothermic ischemia (n = 6), ischemia induced under 30 degrees C body temperature. Cerebral intracellular pH was measured using in vivo 31P NMR spectroscopy over 7 days. Neuronal injury and astrocytic reactivity were evaluated using hematoxylin and eosin staining, and immunoreactivity to glial fibrillary acidic protein, respectively. Normothermic animals revealed significant alkalosis (P less than 0.01) at 48 h after ischemia compared to the pre-ischemic value. No significant intracellular pH change was detected after ischemia in the hypothermic group. Ischemic neuronal injury was prevented in the hypothermic animals, compared to the severe neuronal injury found in the normothermic animals (P less than 0.01). The marked astrocytosis of normothermic animals was significantly inhibited in the hypothermic animals (P less than 0.01). Our data indicate, that hypothermia significantly inhibits neuronal injury as well as post-ischemic alkaloids and astrocytosis, induced by 15 min of forebrain ischemia in the rat.


Muscle pathology correlates with permanent weakness in hypokalemic periodic paralysis: a case report.

We present a morphological follow-up in a case of familial hypokalemic periodic paralysis with progressive weakness. At age 12 years muscle biopsy revealed mild vacuolar changes. Seventeen years later, after the patient had developed permanent weakness, light and electron microscopy disclosed tubular aggregates in about 15% of the fibers and medium-grade myopathic alterations. We describe correlation of muscle pathology with permanent weakness in hypokalemic periodic paralysis.


[The effect of sodium-4-[2-(2,3-dimethyl-4-[1-(4- isobutylphenyl)ethoxy]benzolamino)phenoxy] butyrate (ONO-3805) and antiandrogenic agents on the rat accessory sex organs].

The effects of sodium-4-[2-(2,3-dimethyl-4-[1-(4-isobutylphenyl) ethoxy]benzolamino)phenoxy]butyrate (ONO-3805), newly developed as a non-steroidal 5 alpha-reductase inhibitor, on rat accessory sex organs was compared with chlormadinone acetate (CMA) and non-steroidal antiandrogen, flutamide (FLU). ONO-3805 demonstrated excellent antiandrogenic activity which was similar to that of CMA in reducing the accessory sex organ weight. Moreover this agent did not show any significant effects on the serum testosterone level, but CMA and FLU changed it significantly. We previously reported the androgenic activity of antiandrogen (CMA, FLU, AA560, cyproterone acetate, medroxy progesterone, estrogen), but ONO-3805 is free of androgenic activity.


[Papillary adenocarcinoma of the prostate with good response to antiandrogen therapy and delayed radiotherapy: a case report].

A 76-year-old man complained of initial hematuria and dysuria. Right lobe of the prostate was elastic soft, enlarged and hypoechoic. The serum levels of PA and gamma-Sm were abnormally high. Prostatic biopsy showed papillary adenocarcinoma which was stained by prostatic specific antigen (PA). The cancer was clinically diagnosed as stage C. Antiandrogen therapy was performed. After three months, the prostatic tumor markers decreased to the normal range and the tumor reduced in size without any findings of metastasis on CT scan. Because prostatic biopsy showed viable cancer cells, radiotherapy was added. After six months, the tumor reduced in size without any signs of metastasis on CT scan and prostatic biopsy revealed no viable cancer cells or elevation of the tumor markers. The positive staining for PA and the good response to antiandrogen therapy in our case support the view that papillary adenocarcinoma of the prostate is only a morphologic variant of ordinary prostatic carcinoma (acinous adenocarcinoma).


Papillary cystoadenocarcinoma of the prostate.

We report a papillary cystadenocarcinoma of the prostate found in a 66-year-old man presenting with gross hematuria. The tumor was identified as prostatic in origin because of positive immunohistochemical staining of prostate specific antigen.


[Determination of ER-D5 (estrogen receptor related antigen) in prostatic cancer and its significance].

The existence of an estrogen receptor and various other sex steroid receptors has been confirmed in prostatic cancer. However, the action mechanism of hormone therapy and relationship between the disappearance of hormone response, which is observed in the recurrence of prostatic cancer, and various sex steroid receptors have yet to be identified. The use of monoclonal antibody ERICA is known as a method of detecting the estrogen receptor in the immuno-histological chemistry method. However, this monoclonal antibody is difficult to use on paraffin sections. Therefore, using monoclonal antibody D5, which allows ER-D5 detection of the estrogen receptor related antigen on paraffin sections, we investigated whether or not estrogen receptors are present in the prostatic cancer preparation and studied the survival rate of prostatic cancer as well as relationship with recurrence. Positive ER-D5 results were obtained in 37 of 93 prostatic cancer cases (39.8%) and in all 20 prostatic hypertrophy cases (100%). ER-D5 tended to be more densely stained in prostatic hypertrophy than in prostatic cancer. The survival rate was obviously higher in the ER-D5 positive prostatic cancer cases than in ER-D5 negative cases for a certain period of time after the start of treatment. Despite differences in the histo-differentiation degree and clinical stage of prostatic cancer, the ER-D5 positive percentage did not change and remained between 35% and 45%. In cases where another prostatic cancer preparation was taken because of recurrence of prostatic cancer, there were no ER-D5 positive cases at the time of recurrence.(ABSTRACT TRUNCATED AT 250 WORDS)


[A case of urinary bladder hernia].

A 77-year-old male visited our hospital complaining of swelling in the right scrotal and inguinal region, urinary urgency and difficulty of urination. Physical examination revealed an elastic soft mass in the right inguinal region toward the right scrotum. Rectal examination proved that the prostate was moderately enlarged. Cystogram and urethrography showed hernia of the bladder into the right scrotum and benign prostatic hyperplasia. Urodynamic studies demonstrated organic obstruction in the lower urinary tract. Transurethral resection of the prostate was done for benign prostatic hyperplasia. Later, surgical treatment for the bladder hernia was performed by replacing the bladder into the pelvic cavity and closing the hernial ring without resection of the bladder wall. The bladder hernia proved to be a para-peritoneal type. The postoperative course was uneventful. The previous reports of urinary bladder hernia are reviewed and the incidence, etiology, diagnosis the treatment are discussed.


[A case of giant prostatic calculi].

A 83-year-old man was admitted because of fever and painful swelling of the right scrotal contents on January 23, 1989. Kidney-ureter-bladder X-ray (KUB) showed multiple big prostatic calculi which were compressed postero-laterally by benign prostate hyperplasia (BPH) according to the CT film. UCG film revealed urethral stricture in the anterior urethra. He was diagnosed as having right epididymitis, urethral stricture, BPH and giant prostatic calculi. He was treated with right orchiectomy, urethral bougie and suprapubic prostatectomy with removal of the calculi. The total weight of the prostatic calculi was 28 g of consisted of 125 pieces. The postoperative course was uneventful and two years after the operation he has been well and free from evident disease.


[A multicenter, fixed-flexible dose study of terazosin hydrochloride in the treatment of symptomatic benign prostatic hypertrophy].

In this study the multicenter, fixed-flexible dose regimen was taken to evaluate the effective dose range of Terazosin for the treatment of micturition disturbance in benign prostatic hypertrophy (BPH) and to clarify the characteristics of patients who are more responsive to Terazosin therapy. After a 1-week washout (placebo) the first two weeks 1 mg/day of Terazosin was administered, then depending on efficacy of subjective symptoms, Terazosin doses were increased up to 2 mg/day and 4 mg/day at intervals of two weeks. After six weeks the final efficacy and safety were assessed. The subjective symptom improvement rate was 18.5% by 1 mg/day, 55.6% by 2 mg/day and 65.4% by 4 mg/day cumulatively. The objective symptom improvement rate were 13.2% by 1 mg/day, 42.1% by 2 mg/day and 50.0% by 4 mg/day cumulatively. The global improvement rate was 14.5% by 1 mg/day, 50.0% by 2 mg/day and 61.8% by 4 mg/day cumulatively. The patients who had a higher subjective symptom score in the lead-in period were more improved rather than those who had a lower score. In objective symptoms, voided volume, maximum flow rate (MFR), MFR nomogram score and average flow rate improved and the ratio of residual urine volume decreased. There was no relationship between clinical improvement on either subjective or objective symptoms and prostatic weight. Adverse reactions, such as dizziness, vertigo, tinnitus, nausea and blurred vision; were seen in 10 cases. In conclusion Terazosin was effective and well tolerated for the treatment of patients who had micturition disturbance with BPH in the dose range of 2 to 4 mg/day.


[Clinical evaluation of oral ofloxacin in 3-day therapy for transurethral resection of the prostate].

The effectiveness of oral administration of an antibacterial alone was compared with that of intravenous administration of an antibiotic in patients with benign prostatic hyperplasia undergoing transurethral resection of the prostate (TUR-P). In group A (23 patients), surgery was carried out by administering 200 mg of ofloxacin (OFLX) alone, during the day before surgery and in the morning of the day of surgery and three times a day from the morning after surgery for two days, making the total dose 1,800 mg, and total period of administration 3 days. In group B (22 patients), 1 g of Cefotetan (CTT) was intravenously infused twice a day for 3 days from the operative day, making the total dose 6 g. The patients in both groups were given no other antibacterials or antibiotics. The maximum body temperature was recorded for post-operative 2 weeks. Negativization of bacteria by bacterial culture in the urine and disappearance of the pyria, were followed up on an outpatient bases. The mean maximum body temperature up to post-operative week 2 was 37.1 +/- 0.3 degrees C and 37.0 +/- 0.3 degrees C in groups A and B, respectively. The mean number of days required until negativization of bacteria in the urine was 41.5 +/- 38.9 days and 43.3 +/- 24.9 days in groups A and B, respectively. The mean number of days required until disappearance of the pyuria was 76.5 +/- 23.0 days and 68.2 +/- 17.9 days in groups A and B, respectively. No significant differences were noted by Student's t-test between groups A and B.(ABSTRACT TRUNCATED AT 250 WORDS)


Development of solid-phase enzyme-linked immunosorbent assays for the determination of epidermal growth factor receptor and pp60c-src tyrosine protein kinase activity.

Solid-phase ELISAs for the determination of EGF receptor (EGF-R) and pp60c-src tyrosine protein kinase activity are described. The methods were developed and optimized using purified recombinant EGF-R intracellular domain (ICD) and pp60c-src tyrosine protein kinases. A standardized assay that utilizes poly (GluNa-Tyr)4:1 as substrate and a monoclonal antiphosphotyrosine antibody for detection is described. Assay conditions for both enzymes were optimized with respect to substrate and ELISA plate-coating condition, divalent metal ion preferences, enzyme concentration, apparent kinetic constants for ATP, and reaction linearity. Following standardization, a number of reference tyrosine protein kinase inhibitors were tested in the ELISAs and compared to results obtained using solution-phase radioactive tyrosine protein kinase assays, which are based on the transfer of 32P from [gamma-32P]ATP to synthetic substrate. To enable a comprehensive comparison, IC50 values obtained in the ELISA were compared with values obtained in radioactive assays using both the holo-EGF-R and EGF-R ICD kinases. No substantial qualitative differences between these assays were seen. For many routine tyrosine protein kinase assays, semiquantitative or qualitative measurement of TPK activity is adequate. For such purposes, the ELISAs would be an attractive alternative to radioactive assays.


Exogenous primer-independent cDNA synthesis with commercial reverse transcriptase preparations on plant virus RNA templates.

Upon reverse transcription and cloning manipulations with virion RNAs of several plant viruses, namely beet yellows virus, brome mosaic virus, and potato virus X, we came across a significant background synthesis of cDNA on the virion RNA template in vitro independent of exogenous primers added. When tested with beet yellow virus RNA template, several commercial preparations of avian myeloblastosis virus (AMV) reverse transcriptase showed the background activity monitored by the [alpha-32P]dNTP incorporation in vitro, while the enzyme from murine moloney leukemia virus (MMLV) was found strictly exogenous-primer-dependent. To detect possible nucleic acid contaminations in reverse transcriptase, the enzyme preparations from several commercial sources were incubated with [gamma-32P]ATP and polynucleotide kinase. The labeled material from AMV reverse transcriptase preparations comigrated with a tRNA marker in polyacrylamide gels and was found to be RNase-sensitive. The MMLV reverse transcriptase preparations were free from such a contamination. These results indicate that the exogenous-primer-independent cDNA synthesis by some AMV reverse transcriptases could be due to a contaminating tRNA (or its low-molecular-weight degradation products) serving as an endogenous primer.


Centrifugal elutriation of large fragile cells: isolation of RNA from fixed embryonic blastomeres.

In order to analyze the RNA populations present in different cells of very early embryos, we have developed a protocol to purify these large blastomeres using counterflow centrifugal elutriation (CCE). This procedure employs ethanol fixation to stabilize the cells against shear forces encountered during CCE. Using this method, we fractionated the three different blastomere types of the 16-cell sea urchin embryo, the micromeres, mesomeres, and macromeres, achieving 96, 94, and 96% mean purities, respectively. We show here that intact RNA is recovered with equal efficiency from each blastomere preparation. Using this method, we have identified several RNAs that are distributed non-uniformly among these cells.


Molecular weights of glycosylated and nonglycosylated forms of recombinant human stem cell factor determined by low-angle laser light scattering.

The molecular weight of recombinant human stem cell factor (SCF) was determined using a low-angle laser light scattering combined with a differential refractometer and a uv detector. The protein samples were applied to these detectors through a gel filtration column by a high-performance liquid chromatographic pump. The Chinese hamster ovary (CHO) cell-derived SCF gave a molecular weight of 53,000 for the entire molecule and 35,000 for the protein moiety only at pH 7.0, indicating that the CHO cell-derived protein is glycosylated by 34%. Since the molecular weight of the polypeptide is 18,600, the results demonstrate that the CHO cell-derived SCF forms a dimer. The molecular weight of Escherichia coli-derived SCF was determined to be 39,000, similar to the above value (35,000). Essentially identical molecular weights were obtained at pH 3.0, indicating no dissociation of the dimer.


Management of an outbreak of Norwegian scabies.

An outbreak of Norwegian scabies in a 170-bed acute care hospital was controlled through an organized plan for delivering treatment to those affected: four patients, 50 staff members, and 14 family members of staff members. Health departments in two counties were notified and found four additional cases in the long-term care facility at which the index patient lived. Contact isolation was used for the index patient and any other patients with nosocomial scabies. Staff members infested with Sarcoptes scabiei were released from work until they had been treated with lindane. Staff members who had been in contact with infested persons and staff members' families were treated prophylactically with lindane. This aggressive treatment plan resulted in rapid resolution of the outbreak.


Neutrophil activation associated with increased neutrophil acyloxyacyl hydrolase activity during inflammation in cattle.

Acyloxyacyl hydrolase (AOAH) is a lysosomal enzyme found in neutrophils and macrophages that acts to partially deacylate the lipid-A component of the endotoxin of gram-negative bacteria rendering it less toxic, yet maintaining much of its immunostimulatory potential. We have found that the activity of neutrophil AOAH per cell increased during localized inflammation. The purpose of this study was to determine the mechanism(s) responsible for these increases in neutrophil AOAH activity. Because changes in neutrophil maturity commonly are associated with inflammation, intravascular infusion of purified gram-negative bacterial lipopolysaccharide and SC injection of bovine recombinant granulocyte colony-stimulating factor was used to induce large numbers of circulating immature neutrophils. Immature neutrophils were found to have AOAH activity equal to that of mature cells; however, when neutrophils were stimulated in vitro with known activators, AOAH activity of activated cells was more than that of unstimulated cells. The increase in AOAH activity was inversely related to prestimulation activity. Increases in AOAH activity after neutrophil activation were not a result of de novo synthesis of the enzyme, because cycloheximide did not prevent activation-induced increases in activity.


Culture and initial characterization of the secretory response of neoplastic cat mast cells.

Mast cells isolated from feline splenic mastocytomas were cultured to study their structural and functional properties. Isolated cells from various cats were grown as monolayer cultures for a mean of 56 days (range, 30 to 76 days). Cat mast cells released allergic mediators in response to compound 48/80, anti-cat serum antibodies, and concanavalin A. On the basis of the finding that secretion from cat mast cells was stimulated by anti-cat serum antibodies and concanavalin A, these cells contain surface-bound immunoglobulins. The presence of mast cell-sensitizing antibodies has been suspected in cats, but never before directly demonstrated. Cultured cat mast cells have cytochemical and functional characteristics common to connective tissue-type mast cells and provide one of the few non-rodent models of cultured cells for the study of this type of mast cell.