Measurement of gastric bicarbonate secretion in the human stomach: different methods produce discordant results. Human gastric bicarbonate secretion has been measured by back-titration, from pH and pressure of carbon dioxide (PCO2) determinations (using the Henderson-Hasselbalch formula), and from equations based on gastric juice osmolality and [H+] (osmolality-[H+] method). Since these methods show large quantitative differences in their estimations of gastric bicarbonate secretion, we examined each to define the reasons for these discrepancies and establish guidelines for future work in this area. Bicarbonate recovery from 'non-parietal' secretions (0 to 80 mM HCO3) reacting with 'pure parietal secretion' (160 mM HCl) was studied both in vitro and in the pylorus-occluded healthy human stomach during acid suppression, exogenous acidification, and pentagastrin stimulation. The pH/PCO2 method estimated HCO3- accurately under anaerobic conditions in vitro, whereas the osmolality-[H+] method (with correction factors for osmolality incorporated by us) was accurate under aerobic conditions. In the acid-suppressed stomach back-titration was significantly more accurate than the pH/PCO2 method. In the exogenously acidified and pentagastrin-stimulated stomachs the pH/PCO2 method underestimated bicarbonates, and the osmolality-[H+] method was spuriously elevated in the low range and diminished at high bicarbonate concentrations. Estimates of 'basal' bicarbonate secretion (at zero added bicarbonate) were severalfold higher by the osmolality-[H+] method (5.26 +/- 0.33 mmol/h) than by the pH/PCO2 method (1.20 +/- 0.23 mmol/h) or back-titration (0.65 +/- 0.14 mmol/h). In conclusion, gastric bicarbonate was determined most correctly by back-titration in the acid-suppressed stomach, whereas measurement of bicarbonate in the acid-secreting stomach was not accurate with any method.


Restorative proctocolectomy--the pouch operation: good or bad? Restorative proctocolectomy is now the procedure of choice for patients requiring surgery for ulcerative colitis and some for familial adenomatosis. The indications for operation are being refined, and technically the procedure has been simplified. Although it avoids ileostomy, it does not restore normality. Stool frequency, continence problems, and pouchitis can all spoil a good result. In spite of these shortcomings, it represents a major advance.


Tumours of the liver.

Tumours of the liver. One of the major debates in hepatocellular carcinogenesis at present is whether the hepatitis-B and -C viruses are directly carcinogenic or exert their effect indirectly by causing chronic necro-inflammatory hepatic disease, which in turn is responsible for malignant transformation of hepatocytes. This debate has been fueled by the observation that hepatitis C virus is a single-stranded RNA virus with no precedent for inducing cancer but with a marked propensity to cause chronic necro-inflammatory hepatic disease and by the findings in Chisari's transgenic mouse model, which suggest that severe and prolonged hepatocellular injury per se induces a proliferative response that progresses to tumour formation. Recent reports of a guanine to thymine mutation of the third base of codon 249 of the tumour suppressor gene, p53, in 50% of patients with hepatocellular carcinoma in regions of high aflatoxin exposure, and mutagenic experiments showing that aflatoxin B1 binds particularly to guanine residues in G-C-rich domains and that codon 249 is a preferred target have suggested a mechanism whereby aflatoxin might induce malignant transformation.


The effect of alpha-adrenoceptor stimulation and blockade on the static urethral sphincter function in healthy females. The influence of noradrenaline and prazosin on the urethral closure function was evaluated in 10 healthy female volunteers. Measurements of pressure at varying cross sectional areas were carried out at the bladder neck, in the high pressure zone, and in the distal urethra. Prazosin reduced the static pressure, predominantly in the midportion of the urethra, whereas noradrenaline caused no significant pressure change. The urethral resistance to dilatation and the hysteresis were unaffected by the two agents. It is suggested that the response to prazosin is related to decreased activity of the urethral smooth as well as of the striated muscles, the latter as a result of a reduced somatomotor output from the central nervous system.


Changes in the noradrenaline content of the female rat urinary bladder after transient acute overdistension. The effect of transient acute overdistension produced by forced diuresis and outflow occlusion was examined in female Sprague-Dawley rats. Their bladders were distented for three hours and studied at selected intervals during the postoperative period using high performance liquid chromatography (HPLC) for the exact measurement of tissue noradrenaline. Biopsies were taken from the bladder dome and anterior body 10 hours, 2 days, 7 days and 21 days after overdistension. A reduction in NA concentration was found in both the dome and anterior body at 2 to 21 days after distension. The results demonstrated at least partial transient damage to the adrenergic nerves containing NA. The primary success of the distension therapy used to treat bladder instability may be due to this injury.


Atrial natriuretic peptide and parathyroid hormone (1-84) in relation to noradrenaline induced changes in blood pressure in uraemic and healthy subjects. In order to evaluate the hormonal regulation of blood pressure (BP) in uraemia 12 patients on chronic maintenance dialysis and 14 healthy controls were studied. BP and plasma concentrations of atrial natriuretic peptide (ANP), cyclic 3',5'-guanosine monophosphate (cGMP), and intact parathyroid hormone (PTH(1-84)) were determined before, during, and after a 60 min noradrenaline infusion 0.1 micrograms kg-1 body wt. min-1. Mean BP increased to the same extent in the uraemic patients (median 15 mmHg, range 6-25 mmHg) as in the controls (12 mmHg, 5-25 mmHg). ANP increased during noradrenaline infusion both in patients (7.2 to 8.3 pmol/l, medians, p < 0.01) and in controls (4.4 to 6.0 pmol/l, p < 0.01), and so did cGMP (patients: 31.6 to 35.9 nmol/l, p < 0.05; controls: 6.6 to 8.7 nmol/l, p < 0.01). PTH(1-84) was higher in the uraemic patients than in the controls, but was unchanged during noradrenaline infusion in both groups. Correlation analyses gave no evidence of a direct relation between BP and ANP, but basal PTH(1-84) was negatively correlated to basal mean BP in the patients (rho = -0.615, p < 0.05), but not in the controls. In conclusion, noradrenaline induced similar elevations of BP in dialysis patients as in healthy controls despite elevated ANP and PTH(1-84) in the patients, and ANP release was stimulated in both groups. PTH(1-84) may participate in blood pressure regulation in uraemic patients.


Congenital cystic mesoblastic nephroma: a rare cystic renal tumour of childhood. Case report. A 3-month-old baby boy presented with a right-sided abdominal mass that was shown on radiographic and ultrasonographic examination to be cystic and within the kidney. Histological examination of the right nephrectomy specimen showed it to be a congenital cystic mesoblastic nephroma. The patient made an uneventful recovery and there were no signs of recurrence eight years later. Though this tumour is extremely rare it should be considered as a differential diagnosis in infancy as its prognosis and treatment are different from those of other tumours.


[New receptor ligands for single-photon emission tomography of the brain]. Emission computer tomography offers a unique opportunity to examine functional receptor processes. With the help of specific exogenous radioligands for the various receptor systems of the brain it is possible to visually depict receptor-neurotransmitter interactions, metabolic consequences and changes in diseased conditions. Although radioligands for application in positron emission tomography (PET) are relatively numerous, those for single photon emission tomography (SPECT) are markedly fewer. However, the general availability of the SPECT method gives the SPECT radioligands a special significance. The known SPECT ligands for brain receptors are introduced and discussed. While for certain receptor systems (eg glutaminergic and opiate) no generally useful substances are available, in the case of other receptors (eg cholinergic, serotonergic, dopaminergic and the benzodiazepine system) certain molecules are undergoing clinical evaluation. Iomazenil, a benzodiazepine antagonist, is on the verge of clinical application. Particularly in the case of partial epilepsy, and also possibly in schizophrenia and Alzheimer's disease, this SPECT radioligand is seen as a clear advance in diagnostic methodology. In the light of Iomazenil studies it has been shown how the reproducible analysis of SPECT images can be improved using the Talairach atlas. Finally, the question of whether and how SPECT methodology could be used to determine receptor affinity constant and receptor density is discussed. It is apparent that, under certain conditions (eg., low unspecific binding, slow kinetic etc.), the determination of affinity constants should be feasible using this approach.


[Infectious hemorrhagic disease of rabbits in the Bern pathology institute (1988-90): seasonal and regional distribution and histopathological findings]. Between 1988 and 1990 we have investigated 45 cases of the rabbit viral haemorrhagic disease (VHD) at our institute in order to ascertain: 1) both monthly and regional distribution of the disease: 2) the histopathological patterns of the disease. The VHD cases were particularly frequent between October and January and between May and August. They occurred in 7 cantons. The panlobular hepatocytic necrosis was observed in 80% of the cases, whereas in the remaining 20% the necrosis occurred in the periportal region. In 80% of the animals the spleen blood vessels were dilated with perivascular fibrin accumulation indicative of hyperpermeability. In two cases haemoglobin cages were observed in the kidney tubules.


Transport of proteins across the endoplasmic reticulum membrane. The biosynthesis of many eukaryotic proteins requires their transport across the endoplasmic reticulum (ER) membrane. The process can be divided into two phases: (i) a targeting cycle, during which, by virtue of their signal sequences, nascent polypeptides are directed to translocation sites in the ER and (ii) the actual transfer of proteins across the membrane. The first phase has been well characterized, whereas the latter until recently was completely unresolved. Key components of the translocation apparatus have now been identified and it seems likely that they form a protein-conducting channel in the ER membrane. The transport process is similar to the process of protein export in bacteria.


Response properties of upper cervical spinothalamic neurons in cats. A possible explanation for the unusual sensory symptoms associated with upper cervical lesions in humans. Lesions in the foramen magnum and upper cervical spinal cord often cause an unusual array of sensory changes and atrophic weakness, primarily involving the ipsilateral forelimb. Furthermore, small midline myelotomies performed at C1 often lead to widespread analgesia covering most of the body in patients with chronic pain. These observations challenge physicians' understanding of anatomy and physiology in the upper cervical region. Using single cell recording techniques the authors have shown that spinothalamic neurons in the second cervical segment of cats have complex response properties, often responding to stimuli throughout the body. These findings together with a review of clinical and basic science literature are used to provide explanations for the unusual signs and symptoms observed in patients with upper cervical and foramen magnum lesions.


Chromosomal assignments of mouse connexin genes, coding for gap junctional proteins, by somatic cell hybridization. The connexin genes Cx31 and Cx45 coding for proteins of gap junctional subunits have been assigned to mouse chromosomes 4 and 11 by Southern blot hybridization of specific gene probes to DNA from mouse x Chinese hamster somatic cell hybrids. In addition, our results confirm the recent assignment of mouse connexin genes Cx26, Cx32, Cx37, Cx40, Cx43, and Cx46 to mouse chromosomes 14, X, 4, 3, 10, and 14, respectively, by analysis of interspecific backcrosses and by somatic cell hybridization. Our assignment of the Cx31 gene to mouse chromosome 4 locates the fourth connexin gene on this mouse chromosome to which the genes for Cx31.1, Cx37, and Cx30.3 have previously been assigned. Interestingly three of them (coding for Cx31, Cx31.1, and Cx30.3) are preferentially expressed in skin. Possibly some of the connexin genes clustered on mouse chromosome 4 may be regulated coordinately.


Direct selection of hepatoma cell variants deficient in alpha 1-antitrypsin gene expression. Expression plasmids containing the human alpha 1-antitrypsin (alpha 1 AT) promoter fused to either adenine phosphoribosyltransferase (aprt) or xanthine-guanine phosphoribosyltransferase (gpt) coding sequences were sequentially introduced into APRT- HPRT- rat hepatoma cells. Stable transfectants expressing both transgenes were isolated and characterized. Nonexpressing variants were subsequently obtained by selecting against expression of one or both transgenes. Variants isolated by selecting against expression of either transgene alone generally displayed deficiency phenotypes in cis, as only three of 20 clones tested were affected for expression of alpha 1AT mRNA. In contrast, double selection yielded predominantly trans effects: 12 of 14 lines tested showed impaired ability to express their chromosomal alpha 1AT genes. Furthermore, expression of several other liver genes, including the gene encoding the HNF-1 trans-activator, was repressed in many of the variant lines. Thus, double selection using chimeric transgenes is a useful approach for generating variant cell lines deficient in expression of specific mammalian genes.


[Cystosarcoma phylloides in adolescent women]. The authors describe four cases of juvenile fibroadenoma (benign form of cystosarcoma phyllodes) in pubertal girls aged 11-15 years. They reflect on the variable behaviour of this rare tumour and differentiate the relations between fibroadenoma, virginal hypertrophy of the breast, juvenile fibroadenoma, giant fibroadenoma and cystosarcoma phyllodes. A dominant position is held by surgical treatment which is strictly individualized. The authors recommend, similarly as many other authors, an extensive excision of the tumour with a safety margin and in case of repeated relapses or the very rare malignant form which frequently behaves as a benign tumour. The operation is made in the submammary groove or on the lateral margin of the breast which ensures an excellent cosmetic effect. The authors do not consider excision of superfluous skin or reconstruction of the breast necessary with regard to the elastic properties of the skin and the capacity of the mammary gland to develop.


Update on solid tumor management in childhood. Surgery will continue to play an important role in the multidisciplinary treatment of solid tumors in children. However, as the complexity of therapy increases, it is imperative that the pediatric surgeon become familiar with new radiation therapy techniques, new chemotherapeutic agents, and the advances that are being made in immunodiagnosis, immunotherapy, and molecular biology. New markers for various tumors are being discovered and have therapeutic and prognostic value that may have a significant impact on surgical therapy. New protocols are being developed within the various cooperative research groups, and the surgeon must participate actively, so that surgical questions of importance can be answered and so that adequate tissue specimens can be obtained for future investigations.


[The differences between interstitial pneumonia following whole-body irradiation and radiation-induced pneumonitis]. Clinical observations and experimental results suggest that radiation pneumonitis occurring after localised lung irradiation and interstitial pneumonia occurring after localised lung irradiation and interstitial pneumonia occurring after total body irradiation and bone marrow transplantation (no matter whether or not associated with cytomegalovirus infection) are two different pathological entities. They differ with regard to latency, pathogenesis, dose dependence, and probably dose distribution. Based on these considerations some recommendations on the planning of total body irradiation are made.


Results of radiotherapy with curative intent in non-small cell lung cancer. An analysis of 130 patients. Between 1970 and 1985, 130 patients were irradiated with curative intent at the Center of Oncology in Kraków. The histological diagnosis was squamous-cell carcinoma in 60.8% of patients, adenocarcinoma in 25.4% of patients and other non-small cell cancer in 13.8% of patients. Out of 130 irradiated patients 21.5% refused surgery, 26.2% were inoperable for medical reasons, and 52.3% had unresectable tumors. According to the UICC TNM 1987 classification, 62 (47.7%) patients had early (stages I and II) disease. The remaining 68 (52.3%) patients had stage III A cancer. Additional criteria for patients selection to radiotherapy with curative intent were: Karnofsky performance status > or = 50, and no respiratory insufficiency. All patients were treated with megavoltage radiation. Patients with stage I were treated by three isocentric beams. Tumor dose was 6000 cGy in 24 fractions over five weeks. In patients with stage II and III A disease the radiotherapy was started with two parallel opposed beams encompassing primary lesion and mediastinum. The dose of 4000 cGy was given in 20 fractions over four weeks, followed by a "boost" dose of 2000 cGy in the fractions over two weeks, delivered with three isocentric beams. 54% of patients were disease-free at the twelfth month, 24.6% at the 36th month, and 18.5% of patients survived five years without evidence of cancer. A significantly better survival has been observed in patients with stages I and II, with Karnofsky performance status > or = 70, and with complete radiological regression eight weeks after radiation therapy. The main cause of failure of the treatment were distant metastases.


Benign fibrosing disease at the hepatic confluence mimicking Klatskin tumors. Hilar obstructions remain a challenge with regard to diagnosis and treatment. In the period from 1984 to 1990, 82 patients underwent resective surgery under the presumptive diagnosis of hilar cholangiocarcinoma (Klatskin tumor). The diagnosis was based on the combined appearances on direct cholangiography and ultrasonography in all cases, with the use of various other imaging modalities in some cases. The perioperative findings from an experienced surgical team were usually thought to be compatible with bile duct carcinoma. However, histologic examination of the resected specimens revealed benign fibrosing or localized sclerosing lesions in 11 patients (13.4%). The current state of diagnostic imaging fails as yet to discriminate reliably between benign and malignant hilar lesions. Whereas the immediate therapeutic consequences may be equal (resection followed by hepaticojejunostomy), the late consequences differ in a major way because benign disease has a much better prognosis. In the presence of suspicious hilar obstruction, operable lesions should not be treated by "palliative" intubational techniques and radiation therapy without a firm diagnosis of malignancy. However, overtreatment (extended liver resection, vascular reconstruction, and liver transplantation) should be avoided as well when a benign lesion has not been ruled out.


[Acute malignant myocarditis and pleurodynia as the manifestation of a probable Coxsackie B viral infection]. The authors describe a 45-year-old female patient suffering from rapid-progressing myocarditis with growing disorders in conduction, repolarization alterations and heart dilatation seen for 3 days. The patient died on the 10th day of the disease which started from fever and catarrhal phenomena. Attack-like pains in the right half of the chest and abdomen, removable by narcotics, were a remarkable disease manifestation. The diagnosis of myocarditis was verified morphologically. The disease may be interpreted as infection with Coxsackie B virus associated with myocarditis and pleurodynia.


[The effect of influenza and para-influenza on the course of ischemic heart disease]. A retrospective analysis was made of 541 case reports of patients with coronary heart disease admitted to the infectious department with the diagnosis of influenza (n-387) and parainfluenza (n-154). In all the patients, the diagnosis was verified serologically with the aid of the hemagglutination inhibition test, with a 4-fold and greater increase of the antibody titer in the serum. In part of the patients, it was confirmed by the above test combined with immunofluorescence in examining rhinopharyngeal smears and in part of influenza patients, it was verified virologically. Exacerbation of CHD was seen in the period of early convalescence in 60% of cases whatever the etiology of viral infection (days 5-13 of the disease). As compared to parainfluenza, influenza provoked the deterioration of CHD significantly more often, especially influenza B (25 and 14.3%). The group at risk for an unfavourable outcome of CHD included patients with influenza and parainfluenza, suffering from postinfarction cardiosclerosis. In this group, exacerbation of CHD was diagnosed in 38.3 and 26.1% of cases, whereas acute myocardial infarction developed in 11.7 and 4.4% of cases. Every second influenza patient and every third parainfluenza patient with acute pneumonia and postinfarction cardiosclerosis demonstrated deterioration of CHD.


[R proteins as a reflection of disorders in the cellular reception mechanisms in kidney diseases]. As many as 47 patients with amyloidosis and 31 with chronic glomerulonephritis (CGN) were examined for the blood serum content of R proteins, chemotactic activity of blood leukocytes and the degree of the inhibitory influence of the autologous serum on chemotaxis. Amyloidosis and CGN were marked by an increase of the level of receptor proteins. A reverse correlation was established between the rise of the content of R proteins and the chemotactic properties of leukocytes. In CGN, the rate of demonstration of the high titers of R proteins and the magnitude of the chemotaxis decline depended on the disease phase (the degree of activity) whereas in amyloidosis, these reactions differed insignificantly in patients with various clinical patterns and stages of renal amyloidosis.


Mouse conceptuses have a limited capacity to elevate the mRNA level of cellular retinoid binding proteins in response to teratogenic doses of retinoic acid. In these studies, we wished to determine the effect of teratogenic doses of retinoic acid on the expression of cellular retinoic acid binding protein I (CRABP-I) mRNA, cellular retinoic acid binding protein II (CRABP-II) mRNA, cellular retinol binding protein I (CRBP-I) mRNA, and cellular retinol binding protein II (CRBP-II) mRNA in mouse conceptuses. Levels of CRABP-II mRNA and CRBP-I mRNA were modestly elevated (2.5-fold and 1.5-fold, respectively) in 9-day gestation conceptuses following treatment of dams with 100 mg/kg b.w. of retinoic acid. These levels were elevated by 6 hr following treatment and remained elevated until 48 and 24 hr, respectively. Two other retinoids, etretinate and retinoyl beta-glucuronide, also moderately elevated CRABP-II mRNA and CRBP-I mRNA levels in conceptuses. In contrast, the levels of CRABP-I mRNA in the conceptuses remained unaffected by treatment with any of these three retinoids. These results demonstrate that conceptuses have a limited capacity to elevate the cellular retinoid binding proteins mRNA levels and presumably the synthesis of their respective proteins in response to high, teratogenic doses of retinoic acid. As a result, an excess of free retinoic acid becomes available to the nuclear retinoic acid receptors, which may lead to inappropriate gene expression and eventual maldevelopment.


[New aspects of asthma therapy].

[New aspects of asthma therapy]. Until recently, optimal bronchodilatation and in particular the inhalation of beta-agonists was considered the mainstay of asthma therapy. Steroids were added only in case of an unsatisfactory treatment effect. Recent epidemiological surveys, revealing an association between asthma mortality and the use of beta-agonists, and well controlled treatment studies suggest a change of concepts. Considering the long-term course of asthma, early suppression of asthmatic airway inflammation appears to be at least as important as an effective bronchodilatation. A prerequisite for a success of an individually taylored pharmacologic treatment is a comprehensive management of the disease, inclusive a continuous patient education.


Effect of tissue factor pathway inhibitor (TFPI) in the HEPTEST assay and in an amidolytic anti factor Xa assay for LMW heparin. Both the HEPTEST and amidolytic anti factor Xa assays are currently being used for heparin activity detection in plasma from patients receiving standard heparin or low molecular weight heparin (LMWH). In this study we have investigated the influence of recombinant and endogenous Tissue Factor Pathway Inhibitor (TFPI) on these assays. The HEPTEST determinations were performed on an ACL 300 R Clottimer using the APTT program which resulted in a longer incubation time with factor Xa than recommended by the manufacturer. rTFPI added to plasma prolonged the HEPTEST clotting time markedly, but had only a little effect in the amidolytic assay. Antibodies against TFPI (anti-TFPI) abolished these effects. The effect of adding rTFPI and Logiparin was additive. When anti-TFPI IgG was added to samples of normal plasma, a statistically significant shortening of the HEPTEST clotting time was seen. When anti-TFPI was added to plasma samples from volunteers who had received Logiparin by subcutaneous or intravenous injection, then the HEPTEST clotting time was shortened considerably. For some samples the clotting time was halved. These experiments show that the HEPTEST clotting time is prolonged not only by heparin-antithrombin III, but also by TFPI released by heparin injection.


The procoagulant response of cytomegalovirus infected endothelial cells. The report describes the effect of an in vitro infection of human umbilical vein endothelial cells with human Cytomegalovirus (CMV). The parameters studied are cellular procoagulant activity, secretion of plasminogen activator inhibitor (PAI-1) and urokinase-type plasminogen activator (u-PA), activation and internalization of factor X and Merocyanine 540 staining. The infection does not result in an increase in PAI-1 and u-PA secretion, but it brings about a procoagulant response, which is relatively rapid compared to the tissue factor mediated response induced by inflammatory mediators. The time course and the coagulation factor dependency suggest a facilitated interaction of coagulation factors on the surface of infected cells. Chromogenic activity measurements after the addition of purified factor X and electron microscopic examination of the cells after addition of colloidal gold-factor X conjugates both point to an internalization of factor X and/or Xa after interaction with the endothelial cell surface. Merocyanine 540 staining suggests that CMV infection leads to membrane perturbations.


Factor VII hyperactivity in chronic dialysis patients. To study factor VII (F VII) hyperactivity in chronic dialysis patients, we measured the plasma levels of F VII activity (F VII c) and antigen (F VII Ag), prothrombin activation fragments 1 + 2 (F1 + 2), thrombin-antithrombin III complexes (TAT), and thrombomodulin in 28 patients on hemodialysis. Marked elevation of F VII c was found in long-term dialysis patients (185 +/- 30%). This hyperactivity was accompanied by both elevation of the F VII Ag level (153 +/- 28%) and enhanced activation of F VII zymogen, expressed as the F VII c/F VII Ag ratio (1.23 +/- 0.23), but pseudocholinesterase activity was decreased. The 6 patients with ischemic heart disease had slightly higher F VII c (200 +/- 25%) than those without ischemic heart disease (181 +/- 30%), although the difference was not significant. Increased F VII c was accompanied by factor Xa hyperactivity (a high plasma F1 + 2 level) in the long-term dialysis patients, but there was no significant elevation of plasma TAT levels when compared with controls matched for age, sex, and the presence or absence of diabetes mellitus. Plasma TAT levels were significantly correlated with plasma thrombomodulin levels, suggesting that thrombin generation in blood as a result of hemodialysis could induce systemic endothelial cell injury.


Fragmin once or twice daily subcutaneously in the treatment of deep venous thrombosis of the leg. In an open, randomised controlled study, 101 patients with phlebographically diagnosed deep vein thrombosis of the leg, not extending into more than two thirds of the femoral vein, were randomised to receive Fragmin (a low molecular weight heparin) administered subcutaneously either once or twice daily in doses of 200 U(anti-FXa)/kg/24h or 100 U(anti-FXa)/kg/12h respectively. Prior to Fragmin unfractionated heparin had been administered by continuous iv infusion for not longer than 24h. Warfarin was administered from the first treatment day. Fragmin was administered for at least 5 days or until the prothrombin complex had been within the therapeutic range for at least 2 days. Patients were kept in bed for the first day but thereafter were ambulant. Phlebography was repeated at 5-7 days. Comparison of the phlebograms revealed a similar improvement (reduction in Marder score) in both groups. There were 5 cases of bleeding: 1 major and 3 minor in the twice daily group and 1 minor bleed in the once daily group. There were no cases of clinical pulmonary embolism. It is concluded that Fragmin, administered as a single daily subcutaneous injection, is effective in the treatment of deep vein thromboses, and offers the advantages of reduced costs, despite higher price of the drug, including reduced nursing time.


Prothrombotic phenotype diversity of human aortic endothelial cells in culture. We have previously demonstrated that human aortic endothelium exhibits morphologic heterogeneity in situ, and this heterogeneity can be reproduced in culture. In this study, we have compared prothrombotic properties of cultured endothelial cells (EC) from areas of human aorta at high risk for atherosclerosis (HP-EC) with EC from areas at low risk (LP-EC). Using paired cultures from the same donors, we have found that the expression of cell surface thrombomodulin (TM)--as measured by the ability to generate activated protein C (APC) from protein C in the presence of thrombin--is relatively reduced on HP-EC compared to LP-EC (respectively, 4.98 +/- 4.43 vs. 5.83 +/- 4.37 pM APC/min/cm2; p = .03, n = 12). Furthermore, HP-EC more efficiently assemble the prothrombinase complex on their cellular surface, resulting in an increased rate of thrombin generation from prothrombin (9.81 +/- 3.10 (HP-EC) vs. 7.96 +/- 3.20 nM thrombin/min/cm2 (LP-EC); p less than .03, n = 7). The combination of reduced TM expression and increased prothrombinase complex assembly on HP-EC suggests a prothrombotic phenotype in these cells. These findings may be important in the pathogenesis of thrombosis associated with atherosclerotic plaques.


Ex vivo activity of heparin is not predictive of blood loss after neutralization by protamine. To study whether the ex vivo activity of heparin and Fraxiparin correlates to and predicts the extent of blood loss induced by the heparins (pre- and post neutralization by protamine), a rat tail transection model and a rabbit ear bleeding model were used. In the rat model heparin (2 mg/kg i.v.) significantly prolonged the bleeding time, while this dose of Fraxiparin had no effect. In the rabbit ear blood loss model, heparin (2 mg/kg i.v.) produced significant increases in blood loss while Fraxiparin (2 mg/kg i.v.) produced approximately 30% of the blood loss induced by heparin. Equigravimetric protamine reduced the heparin-induced blood loss by approximately 50%, however, significant blood loss, thrombin time and Heptest activity remained. Heparin and Fraxiparin (3 mg/kg s.c.) did not cause any increased bleeding. While, all activities of heparin were completely neutralized by protamine, the Heptest activity of Fraxiparin was resistant to neutralization. The ex vivo activity of heparins after neutralization by protamine does not correlate to the extent of blood loss which suggests it may not be necessary to neutralize all ex vivo activities of the heparins to baseline values to be assured that blood loss is reversed.


Stellate cells of aortic intima: II. Arborization of intimal cells in culture. The present study analyzed effects of different cAMP-elevators on cell morphology in primary culture of human intimal and medial cells from grossly normal and atherosclerotic areas. In primary culture of human aortic cells adenylate cyclase activator forskolin and other cAMP elevators induced arborization of cells, i.e. they reversibly changed the shape of cells. This resulted in the formation of thin branching processes and in the concentration of cytoplasm around the nucleus. In the culture, the shape of the arborized cells resembled that of stellate ones detected in the aortic intima in situ. The arborization of cells was accompanied by destruction of myofilaments. Due to cAMP elevators' effect, most of the arborized cells were exhibited in the cultures isolated from the elastic-hyperplastic layer of the intima. The number of arborized cells was significantly less in the cultures isolated from the musculo-elastic layer and still lesser in those isolated from media. We failed to reveal any significant difference in the number of arborized cells cultured from fatty streaks, atherosclerotic plaques and grossly normal aortic areas. Obtained results suggest that the previously revealed polymorphism of human aortic intimal cells may be accounted for by the cell shape transformations underlined by the mechanism similar to that of arborization in vitro.


[Practice observations of cross protection between bovine and porcine herpesviruses]. In many cattle herds in Lower Saxony the serologic IBR/IPV = BHV-1-status is known because since 1988 the bovine herpes virus infection (BHV 1), the infectious rhinotracheitis and pustular vulvovaginitis (IBR/IPV) are being fought. Through extensive investigations in five districts in the Weser-Ems area, it was found that cows infected by bovine herpes virus (BHV 1) are also protected to a high degree against infection by porcine herpes suis virus (SHV 1). An interpretation of the observed cross protection is not yet possible; further research is necessary.


Expression of the immediate early antigens of human cytomegalovirus is responsible for virus proliferation; an intracellular immunization approach. The purpose of this study was to determine the sequence within the immediate early (IE) region of human cytomegalovirus (HCMV) that is the most important in HCMV proliferation. Stable DNA-transfected VA/13 cell lines that express the immediate early antigens (IEA's) of HCMV were established and immunofluorescence staining with patients' antibodies revealed that the transfected DNAs were stable and able to express HCMV-IEA's. A cell line (VA/13-mt) that was transformed with a metallothionein (MT)-promoter was able to express IEA's constantly in 80-90% of cultured cells. Under this condition, VA/13-mt cells produced over 50 times the number of HCMV particles produced by parental VA/13 cells. However, another cell line (VA/13-PX) that was transformed with a fragment deleted at the region of EcoRI H of HCMV DNA did not produce as many HCMV particles as the parental VA/13 cells. These findings indicate that IEA's are responsible for virus proliferation, and that the products from a deleted gene of the IE region may contribute to intracellular immunization.


Abnormalities at different levels of the hypothalamic-pituitary-adrenocortical axis early after stroke. Hypercortisolism is common in stroke patients. The aim of this study was to investigate possible disturbances at different sites within the hypothalamic-pituitary-adrenal axis. We also studied possible associations between hypercortisolism and clinical manifestations of brain dysfunction. Patients with an acute ischemic stroke (n = 16; mean +/- SD age, 71 +/- 11 years) were compared with healthy elderly subjects (n = 9). We performed a short adrenocorticotropic hormone (ACTH) test with 0.25 mg 1-24 ACTH injected intravenously and an overnight dexamethasone suppression test with 1 mg dexamethasone given orally at 11 PM. Serum cortisol levels after dexamethasone at 8 AM were significantly higher in stroke patients (p = 0.003). The area under the curve for the cortisol response to ACTH was elevated in seven (47%) of stroke patients, and the centered cumulative cortisol response was elevated in three (20%) patients. The area under the curve response correlated significantly to the presence of an acute confusional state and male sex in stroke patients (rs = 0.63 and rs = 0.62, respectively; p < 0.05), whereas the centered cumulative cortisol response diminished with increasing age (rs = -0.62; p < 0.05). Postdexamethasone cortisol levels were significantly correlated to the presence of an acute confusional state and to extensive limb paresis (rs = 0.66 and rs = 0.62, respectively; p < 0.05). There are abnormalities in the cortisol axis both at the central level and at the adrenal level early after stroke. Hypercortisolism is closely associated with cognitive disturbances and extensive motor impairment.


Acute endocrine failure after brain death? After brain death, 32 potential organ donors were studied to determine serum and plasma concentrations of hypothalamic-pituitary hormones, thyroid hormones, and cortisol over a period of up to 80 hr. Diagnosis of brain death was established either on the basis of clinical criteria (n = 16) or by angiography (n = 16). While 78% of the organ donors developed diabetes insipidus, none of the circulating hormones of the anterior pituitary gland showed a progressive decline in concentration according to their plasma half-lives. With the exception of arginine vasopressin (AVP), no hormone concentration was found to be subnormal due to the onset of brain death. The subnormal free triiodothyronine (FT3) values in 62% of cases (median FT3 of 2.2 pmol/L within the first 24 hr) and the cortisol concentration of 6.9 micrograms/dl correlate with the frequency of similar findings in patients with severe head injuries. While the adrenocorticotropic hormone (ACTH) concentrations of 10-53 pg/ml remained constant during the study period, thyroid-stimulating hormone (TSH) and human growth hormone (hGH) concentrations showed a 12- and 35-fold increase from baseline values after 30-40 hr. These results suggest that, despite the now generally accepted criteria of brain death, there is still some residual function, and thus also perfusion of the hypothalamic-pituitary neuroendocrine system. This residual function appears to be sufficient to maintain hormonal plasma levels at least in the low reference range in most donors. Hormonal depletion in organ donors subsequent to brain death, as suggested repeatedly in the literature, could not be confirmed. The analysis of serum or plasma concentration patterns of a number of hormonal parameters following brain death does not support the rationale for a routine replacement therapy of total triiodothyronine (TT3) or cortisol to maintain endocrine homeostasis prior to organ harvest. However, dexamethasone therapy may be followed by suppression of the adrenal cortex of the organ donor. In these cases, cortisol substitution may be indicated.


Arboviruses as aetiological agents of encephalitis in the People's Republic of China. A serological study was undertaken to determine the role of arboviruses as etiological agents of encephalitis in the People's Republic of China (PRC). Paired sera were collected during mosquito seasons in 1988-1990 from 614 patients with possible viral encephalitis in 15 regions of PRC and tested for haemagglutination inhibiting antibodies to selected arboviruses. Seroconversions were documented to alphavirus and flavivirus antigens in 13.0 and 18.7% of patients respectively in most of the study areas. No California group seroconversion was detected. The age of alphavirus seroconvertors ranged from 2 months to 32 years and of flavivirus seroconvertors from 6 months to 50 years, with higher numbers in males. Serious central nervous system manifestations were seen more commonly in flavivirus seroconvertors. This study affirms the importance of flavivirus as causative agents of encephalitis in PRC and provides evidence that one or more alphaviruses are causing symptomatic infections with neurological involvement in PRC.


The influence of HLA A-B-DR matching on cytomegalovirus disease after renal transplantation. Evidence that HLA-DR7-matched recipients are more susceptible to cytomegalovirus disease. We examined the prevalence of cytomegalovirus infectious episodes, as defined by clinical, virological, and serological criteria (i.e., CMV disease), in 660 kidney graft recipients; 109 patients (16.5%) developed the disease, and 551 did not. No significant statistical link between CMV disease prevalence and a given HLA-A, -B, or -DR allele was observed. However, patients with HLA-DR7 matched grafts were statistically more frequently found (P < 0.01) in the group of recipients who developed CMV disease as compared with the group who did not develop CMV disease. Furthermore, among patients who developed CMV disease, a significant increase of HLA-DR7 matched over DR7 mismatched patients was noted, whereas no difference between matched and mismatched recipients for the other HLA-DR alleles was found. No difference in the severity of graft failure, often observed during, or immediately after, the CMV episode, was noted between patients matched or mismatched for HLA-DR7. Our data suggest that donor/recipient matching for HLA-DR7 is associated with increased CMV disease.


A prospective study of human herpesvirus-6 infection in renal transplantation. Sixty-five kidney transplant recipients and their (22 living related and 43 cadaveric) donors were studied prospectively to determine the relationship between kidney transplantation and human herpesvirus-6 (HHV-6) infection. The virus isolation from peripheral blood and other tissues and sequential determination of neutralizing antibodies to HHV-6 were performed during 3 months following the transplantation. All of the donors and their recipients examined had neutralizing antibodies to HHV-6 at the time of renal transplantation and the virus was not isolated from them. HHV-6 was isolated from 3 renal tissues (2 living related and 1 cadaveric) obtained during transplant surgery, but not from their blood at that time. HHV-6 viremia occurred in 9 (14%) of the 65 recipients around 2 to 4 weeks after the transplantation. An additional 27 recipients showed a significant rise in the antibody titer. Thus, the infection with HHV-6 was confirmed in 36 (55%) of the 65. These results indicate that the virus is activated in many cases in the early posttransplant period and that HHV-6 establishes in vivo latency in the kidney tissue. There was no correlation between HHV-6 infection and acute rejection or the antirejection prophylaxis.


A new rural focus of cutaneous leishmaniasis caused by Leishmania tropica in Kenya. We have identified a new rural focus of cutaneous leishmaniasis caused by Leishmania tropica in Muruku sublocation, Salama location, Laikipia district, Rift Valley province, Kenya. Based on a few available case histories, previous reports of L. tropica in Kenya indicated a tentative geographical distribution. Recently 6 indigenous Kenyans from the new focus, who had never travelled outside Kenya, developed cutaneous lesions on the face and/or extremities found to contain Leishmania by culture and smear. Most of the patients manifested the typical 'urban' dry sore which grew slowly into a nodule measuring 2 x 1 cm to 9.5 x 3 cm, and after some months formed a central crust surrounded by small satellite papules. After treatment with Pentostam (sodium stibogluconate), about 40% of the sores failed to heal completely, either scarring centrally with fulminating papules at the edges and spreading peripherally, or healing but then recrudescing at the edge of the scar. Stationary-phase promastigotes from culture isolates were analysed by cellulose acetate electrophoresis. Isoenzyme profiles of 6 isolates were compared with those of World Health Organization reference strains using 12 enzyme loci; they were indistinguishable from those of 2 L. tropica reference strains. All 6 case sites lay within a radius of 4 km. Several other suspected cases from the same area are being investigated.


T lymphocyte activation by cytomegalovirus-infected, allogeneic cultured human endothelial cells. Cytomegalovirus, a source of serious complications among immunosuppressed individuals, infects endothelial cells in vivo, and has been epidemiologically associated with allograft rejection and transplantation-associated accelerated arteriosclerosis (TxAA). As the interface between the host immune system and allograft tissues, the endothelium may be of primary importance in mediating pathogenic immune responses to CMV. We have therefore investigated T lymphocyte responses to CMV-infected allogeneic human umbilical vein endothelial cells (HUVE) in vitro. Proliferation assays demonstrated dramatically enhanced responses by CMV-seropositive donor-derived PBMC or purified T cells cocultured with CMV-infected HUVE, as compared with those elicited by noninfected stimulator cells. As determined by limiting dilution analysis of IL-2-producing cells, the frequency of T cells responding to infected HUVE was generally found to exceed by approximately one order of magnitude those responding to uninfected cells. Similar analyses of isolated T cell subsets revealed that these responses (proliferation and IL-2 production) were nearly entirely accountable to the CD4+ fraction. Responses of CD8+ populations, however, varied among donors. The marked activation of CD4+ cells is particularly intriguing since we have shown that CMV-infected HUVE do not express HLA class II antigens. Responses of lymphocytes derived from CMV-seronegative donors were minimal in all assays. These studies show that purified T cells can respond to CMV in the exclusive context of allogeneic endothelial cells. Furthermore, we demonstrate the heretofore undescribed phenomenon of CD4+ T cell activation in the absence of serologically detectable HLA class II antigen. As vascular rejection and TxAA are characterized by subendothelial T cell infiltration, these findings support a role for CMV/endothelial interaction in their pathogenesis.


Development of a simple indirect enzyme-linked immunosorbent assay for the detection of immunoglobulin M antibody in serum from patients following an outbreak of chikungunya virus infection in Yangon, Myanmar. During 1984, 1548 children were admitted to the Yangon [Rangoon] Children's Hospital in Myanmar [Burma] with haemorrhagic fever. No evidence of recent dengue infection was found in 577 of the 803 children from whom paired sera were obtained, raising the possibility of reappearance of Chikungunya virus infection in Myanmar. An enzyme-linked immunosorbent assay (ELISA) for the detection of anti-Chikungunya virus immunoglobulin M (IgM) antibody was prepared and standardized using only reagents which are commercially available or which could be prepared without the use of sophisticated equipment. While there was 90% agreement between haemagglutination inhibition (HI) tests and the IgM ELISA in the diagnosis of acute Chikungunya virus infections, 12 additional patients with stationary anti-Chikungunya virus HI antibody titres could be identified as having acute Chikungunya infections using the ELISA. Furthermore, the ELISA could identify twice as many patients (31/103) at the time of admission to hospital as the HI test (15/103). There was no false positive IgM reaction with the ELISA which could be attributed to the presence of rheumatoid factor. Using the test, 103 of a sample of 163 children who presented to the Yangon Children's Hospital with fever/haemorrhagic fever were diagnosed as Chikungunya patients, 4 had possible dual Chikungunya and dengue infections, 16 had dengue, 30 had neither Chikungunya nor dengue infections, and a definitive diagnosis could not be made for 10 patients. Routine use of the ELISA would alert authorities to future outbreaks of Chikungunya virus infection and avoid admission to hospital of patients with a non-life-threatening viral disease.


Yeast SNF/SWI transcriptional activators and the SPT/SIN chromatin connection. Genetic studies of many diversely regulated genes in the yeast Saccharomyces cerevisiae have identified two groups of genes with global functions in transcription. The first group comprises five SNF and SWI genes required for transcriptional activation. The other group, containing SPT and SIN genes, was identified by suppressor analysis and includes genes that encode histones. Recent evidence suggests that these SNF/SWI and SPT/SIN genes control transcription via effects on chromatin. SNF2/SWI2 sequence homologues have been identified in many organisms, suggesting that the SNF/SWI and SPT/SIN functions are conserved throughout eukaryotes.


Mls genes and self-superantigens.

Mls genes and self-superantigens. The Mls gene products, which have long been known for their potent T-cell stimulatory function, have recently come of age through two pivotal discoveries, revealing that they act as superantigens and originate from retroviruses. In addition, recent experiments suggest that two retroviruses, the murine B-type mammary tumor virus and the human lentivirus HIV, make use of the T-cell stimulatory capacity of a virally encoded superantigen for facilitating viral replication.


The fall and rise of neuronal alpha-bungarotoxin binding proteins. Although neuronal [125I]-alpha-bungarotoxin binding proteins are similar in many respects to muscle nicotinic acetylcholine receptors, their functional significance has eluded researchers for the past fifteen years. Over this period, their status became increasingly doubtful, as almost all attempts failed to demonstrate that alpha-bungarotoxin could block neuronal nicotinic responses. Recently, these enigmatic proteins have been cloned and expressed in oocytes, and have been examined afresh in their native state. As Paul Clarke explains, it is time to recognize neuronal alpha-bungarotoxin binding proteins as distinct members of the nicotinic acetylcholine receptor gene family, even if perhaps they do not function quite like other members.


Small cell carcinoma of the gallbladder: clinical course and response to chemotherapy. Small cell carcinoma of the gall bladder is a rare tumor. The neoplasm is highly lethal, metastasizes early, and may cause death shortly after diagnosis. An oat cell carcinoma of the gallbladder metastatic to the liver and adjacent lymph nodes is described in a 60-year-old male. Partial cholecystectomy was performed followed by aggressive chemotherapy with etoposide and cisplatinum. An 80% reduction in the size of the unexcised tumor was noted over a period of 6 months. The partial response and the relatively long survival of the patient suggest the use of the above protocol for these rare cases.


Evaluation of the sensitivity of cervicography in a consecutive colposcopic series. Cervicography was performed in 606 women referred for colposcopy. Cervigrams were blindly reviewed by two independent readers. The positivity rate at cervicography was high (operator A = 50%, B = 58.8%). The sensitivity for papillomavirus infection (HPV)/cervical intraepithelial neoplasia I (CIN I) (n = 141) was 79.4% for operator A and 80.8% for operator B. The sensitivity for CIN II or more severe lesions (n = 22) was 95.2% and 90.5% for operators A and B, respectively. The positive predictive value for HPV/CIN I or CIN II, or more severe lesions was 36.9% and 6.9% for operator A and 32.1% and 5.3% for operator B, respectively. Interobserver variability was acceptable (kappa = 0.62). Cervicography suspected 27 HPV/CIN I, 1 CIN II and 1 CIN III which showed no cytologic abnormalities. This study confirms that cervicography has a good sensitivity for cervical lesions, but it is based on a selected series, not representative of a screening condition. The combination of cervicography and cytology in screening is presently under evaluation in a prospective study of screened women.


Mixed signet-ring and transitional cell carcinoma of the bladder. Report of a case. A case of transitional cell carcinoma of the bladder displaying prominent signet-ring cell features is reported. Despite combined therapy, the patient died within a few months of the diagnosis, thus confirming the poor prognosis of this tumor. We understand this entity could be considered as an example of neoplastic-associated metaplasia and therefore propose that it should be distinguished from genuine adenocarcinoma.


[Effect of the nature of the surface of disperse silica on its interaction with the reproductive cells and seminal plasma of cattle]. The interaction of various disperse silica of I, II, III kind possessing various structure of surface groups (-OH; -O-CH2-CH2-O-CH2-CH2OH; -O-CH2-CH2-NH2 respectively) was investigated with some above membrane matrix polymers of bovine reproductive cells and seminal plasma (namely the surface proteins and carbohydrate polymers containing the N-acetyl-neuraminic acid (NANA) as terminal residue). Protein binding was preferentially observed for silica surface modified by aminoethoxy--and ethylene glycol groups and depended on concentration of silica in the mixture. It was found that biopolymers containing carbohydrate groups had larger affinity to I than to II or III. The binding value of I-III was 12-16% with respect to plasma proteins. Silicas I and II with -OH-groups on the surface absorb 17-21% N-ANA-containing polymers of bovine seminal plasma.


[Immunogenic proteins of nuclease-sensitive regions of chromatin of the normal rat liver and N-nitrosodiethylamine-induced hepatoma]. The localization of immunogenic proteins of chromatin in the nuclease-hypersensitive regions was investigated by means of polyclonal antibodies against the total chromatin from cells N-nitrosodiethylamine-induced hepatoma of rats and from hepatocytes. The fractions released after digestion by Ca-dependent endonuclease and by exogenous DNasa-1 were considerably enriched with immunogenic determinants. The enrichment of nuclease-hypersensitive sites of chromatin with tumour-associated antigens was observed in experiments with antihepatoma antibodies preincubated with chromatin from the normal liver cells. Higher immunoreactivity was typical of fractions obtained during Ca-dependent endonuclease segregation of the hepatoma chromatin, while products of digestion by exogenous DNasa-1 possessed higher tumour specificity.


Cytomembranous inclusions in myelodysplastic syndrome. A 56-year-old African-American man presented with fever of unknown origin and peripheral blood and bone marrow findings of myelodysplastic syndrome (MDS): refractory anemia with an excess of blasts in transformation that subsequently progressed to acute myeloblastic leukemia (AML). Ultrastructural study of two bone marrow specimens having the findings of MDS revealed frequent, large tubuloreticular structures (TRS) in lymphocytes, plasma cells, macrophages, and endothelial cells. Several cylindrical confronting cisternae (CCC) were present in macrophages and an endothelial cell. Two partially developed CCC were present in a plasma cell. TRS and CCC were not observed in eight subsequent bone marrow specimens obtained during the 9-month course of the AML. This is the first reported occurrence of TRS and CCC in MDS. These inclusions are probably related to an unidentified viral infection or possibly to cytokines released by the dysplastic hematopoietic cells.


Infection of pigs and cattle with bovine viral diarrhoea virus on a farm in England. Deaths within a litter of sucking pigs led to a suspicion of hog cholera, but pestiviruses isolated from both dead and live piglets appeared to be bovine viral diarrhoea virus. Persistent viraemia with bovine viral diarrhoea virus was demonstrated in living littermates and also in a bought-in calf, housed in a nearby pen on the same farm. Only two of the littermates survived, both of which had been virus negative and seropositive from the outset of testing. Porcine and bovine virus isolates grew well in calf testicular cells and were neutralised equally by sera collected at the farm from cattle and pigs. However, a comparison by means of their reactivity to monoclonal antibodies showed that they were similar but not identical, and only the porcine isolates grew well in a porcine kidney cell line.


Genomic variability among globally distributed isolates of equine arteritis virus. Equine arteritis virus (EAV), a non-arthropod borne togavirus, has been shown to have a global distribution. To date, no major antigenic variation has been demonstrated between EAV isolates from different geographic origins. In this study, the genomic RNA of EAV isolates obtained from horses of different breeds in various countries around the world was oligonucleotide fingerprinted. Comparisons of these fingerprints were used to determine the extent of genomic variation among such isolates. Comparisons among isolates from North American horses revealed, for the most part, oligonucleotide homologies of less than 60%. Only 29 of the 98 comparisons revealed greater than 60% oligonucleotide homology. Nonetheless, several comparisons indicated a close epidemiologic relationship between isolates from horses of different breeds located in different states. Though all European isolates were of Standardbred origin and were from horses located in northern European countries, the majority had oligonucleotide homologies of less than 60%. Where oligonucleotide homology was apparent, it was, with one exception, greater than 70%. The two isolates from New Zealand had 93.2% oligonucleotide homology. This is indicative of an extremely close epidemiologic relationship. Comparisons between EAV isolates from around the world revealed oligonucleotide homologies between viruses from North America, Europe and New Zealand. In several instances, this homology was greater than 70% and in one case greater than 80%. No oligonucleotide homology was evident in comparisons involving the virus from South Africa. The high level of genomic conservation between certain EAV isolates of disparate geographic origins may reflect dissemination of the virus associated with the international movement of horses. The extent of genomic variation demonstrated between most of the EAV isolates used in this study confirms the need for further investigation of genomic heterogeneity among strains of this virus before techniques that rely upon nucleic acid hybridization can be effectively applied as diagnostic procedures.


No evidence of vertical transmission of naturally acquired feline immunodeficiency virus infection. A naturally occurring feline immunodeficiency virus (FIV) infection in a closed breeding colony of cats, was studied for a period of 9 months. The colony consisted of 25 adult cats, of which six proved to be infected with FIV as judged by serological examination and virus isolation. In all, 48 kittens were monitored for levels of antibodies against FIV during their first 6 months of life. All the kittens (n = 30) born of FIV-infected queens showed maternal antibodies against FIV, although these declined to undetectable levels by the age of 5 months. Antibodies against FIV were not shown in any of 18 kittens born of FIV-negative queens. An attempt to isolate the virus from 12 kittens between 2 and 6 weeks of age did not succeed. None of the cats in the colony seroconverted during the observation period. In conclusion, neither vertical nor horizontal transmission of FIV infection were demonstrated in the colony during the 9-month investigation period.


Impairment of macrophage function in Mongolian gerbils. Chemiluminescence studies on superoxide generation by phagosomes using opsonized zymosan showed the highest fluorescence in murine splenic macrophages among four different kinds of splenic or peritoneal macrophages from mice or gerbils. Murine splenic macrophages phagocytized two to three times more latex particles than gerbil splenic macrophages, but peritoneal macrophages did not show a significant difference in phagocytic activity between mice and gerbils. Phagocytosis by macrophages was determined by a technique based on measurement of the release of hydrogen peroxide and myeloperoxidase from phagosomes using microspheres conjugated with 3-(p-hydroxyphenyl) propionic acid (HPPA-MS). HPPA is a substrate of lysosomal myeloperoxidase. The fluorescence of HPPA-HPPA-MS produced by phagocytized HPPA-MS was measured with an immunoreaction analysis system (IMRAS), and the enzyme activities of the four different kinds of peritoneal or splenic macrophages from mice and gerbils were compared. All four kinds of macrophages produced HPPA-HPPA-MS in their phagosomes during phagocytosis and murine splenic macrophages showed the highest level of enzyme activity.


Circulating immune complexes in bovine leukemia virus (BLV)-infected cattle. Circulating immune complexes (ICs) were detected in the sera of bovine leukemia virus (BLV)-seropositive cattle. Immune complexes were precipitated in 2.5% polyethylene glycol (PEG) and further dissociated. Bovine leukemia virus antigens, IgG and IgM molecules were detected after solubilization in the presence of sodium dodecyl sulphate, and quantitated by enzyme-linked immunosorbent assay (ELISA) assays. Mean values of IgG and IgM in BLV-containing ICs did not significantly differ from those obtained from ICs originating from BLV-seronegative animals. However, differences were found in the composition of ICs from older BLV-positive animals as compared to those obtained from young animals. The ratio of IgG/IgM was 5.02 in animals aged 5-10 years, while this ratio was 11.66 in animals of less than 5 years of age and 10.19 in controls. This might indicate a possible increase in the contribution of IgM molecules to the structural composition of ICs in BLV-infected cattle as related to age or stage of infection.


Marek's disease virus-transformed chicken T-cell lines respond to lymphokines. Current assays for chicken interleukin-2 (IL-2) utilize mitogen-activated lymphocytes. However, very high inter-assay variability and sporadic high background proliferation limit their usefulness. In view of the above, several Marek's disease virus (MDV)-transformed T-cell lines (which grow well in a serum-supplemented medium) were tested for a response to chicken IL-2 when grown in serum-free media. Five of six lines examined showed a dose-dependent proliferative response to chicken T-cell conditioned media. One line, MDCC-CU14, was chosen for further studies. In addition to the tumor cells' dose-dependent responses to semi-purified chicken IL-2, they expressed T-cell activation antigens on the cell surface. Furthermore, the level of surface expression was enhanced on cells provided IL-2. Co-incubation of the tumor cells with monoclonal antibody INN-CH-16 (specific for an antigen on the surface of activated T-cells) and IL-2 resulted in a modulation of lymphokine-induced proliferation. Together, these data suggest that signalling mechanisms in MDV T-cell tumors are intact and that these lines can be used as an assay for chicken T-cell lymphokines. Furthermore, they provide an interesting model for the study of avian and mammalian T-cell transformation. Implications for the study of Marek's disease are also discussed.


Stored blood components contain agents that prime the neutrophil NADPH oxidase through the platelet-activating-factor receptor. Neutrophils (PMNs) initiate production of toxic oxygen metabolites through stimulation of an NADPH oxidase resulting in the reduction of oxygen to superoxide anion (O2-). The activity of this enzyme system may be primed by a variety of compounds. This laboratory investigated the possibility that stored blood components may contain agents which prime the PMN oxidase. At the time of outdate, packed red blood cells, whole blood, and platelet concentrates contained a priming agent which enhanced the PMN NADPH oxidase activity in response to a soluble stimulus, formyl-Met-Leu-Phe, by 2.1- to 2.8-fold. The priming activity was almost completely inhibited by WEB 2170, a specific platelet activating factor antagonist. Fresh plasma or fresh-frozen plasma did not exhibit priming activity. These data suggest that platelet-activating factor-like compounds are generated during the storage of cellular blood components. The presence of these agents in stored blood may suggest a role for specific compounds which prime PMNs and possibly mediate other effects which result in severe complications of transfusion therapy.


[The diagnostic potentials of pirfotekh in cardiovascular diseases]. The authors examined 98 patients with different cardiovascular diseases and 20 practically healthy subjects and established that complex use of pirphotech with 99mTc-pertechnetate gives an objective information on circulatory disorders in the myocardium. Radionuclide ventriculography in the static variant should be carried out early for obtaining adequate information on the state of cardiodynamics.


[Gastric motor-evacuatory function and the rehabilitation of peptic ulcer patients following vagotomy]. A study is presented of the motor-emptying function of the stomach evaluated by dynamic gastric scintigraphy in 43 patients with duodenal ulcer before and after vagotomy. In the early postoperative period 35 patients showed an enhanced emptying of the stomach, 8--a delayed. It was established that the most unfavourable results of vagotomy (gastrostasis, unhealed and relapsing ulcers) were observed in patients with preoperative dyskinesia of the duodenum and registered duodenogastric reflux. This should be considered in selecting the method of operative intervention. B-II gastric resection should be preferred in patients with duodenal dyskinesia.


[Generation of superoxide anion and lipid peroxidation in serum of gout patients]. Alterations in content of lipid peroxidation products, reactions of generation/interception of superoxide anion radicals, content of R-proteins and uric acid were studied in blood serum of 83 patients with gout depending on the disease activity. Rates of lipid peroxidation, estimated by content of thiobarbituric acid-positive products, and of superoxide anion generation as well as content of R-proteins correlated distinctly with activity of the inflammation. At the same time, content of uric acid in blood serum of these patients correlated reversely with content of the lipid peroxidation products at the disease preexacerbation period, while concentration of these products depended on content of total iron.


[The identification of alphaviruses by using cDNA probes complementary to the 3'-terminal of the viral genome]. Venezuelan equine encephalomyelitis, Eastern equine encephalomyelitis and Sindbis viruses, as well as members of various serological groups of alphaviruses were identified using synthesized molecular probes complementary to 3'-terminus of the molecule of the appropriate genomic RNA: 32P-single-stranded kDNA and E. coli-cloned 32P-double-stranded kDNA. A high species-specificity of kDNA-probes was demonstrated. The sensitivity of the method was 1 ng of viral RNA overlayed on the nitrocellulose filter.


[Status of the development of electron spin resonance measurement for the detection of irradiated food]. Electron spin resonance spectroscopy can be used for the detection of irradiation of various groups of foodstuffs. The results of ESR-measurements on irradiated meat and fish and fresh fruit, as well as dried fruit, spices and nuts as performed by the food irradiation laboratory of the German Federal Health Office are summarized in this report. For the detection of irradiated meat and fish, we examined the bones. Using the results from 10 different animal bones, we were able to develop an official method according to the German law section 35 LMBG. A similar routine method for fish will be established in 1992 (at the moment, an intercomparison with German food control laboratories is in progress). Irradiated dried fruit can be identified easily, because unirradiated samples give no ESR-spectra, while irradiated fruit show a partially resolved spectrum, which is caused by radiation induced sugar radicals. Interestingly, the structure of the resulting spectra is not identical for all irradiated species of fruit. We found three different types of ESR-spectra for irradiated dried fruit. Irradiated nutshells show an ESR-spectrum which reveals two additional lines (from cellulose-radicals) beside the main signal, while unirradiated samples show only the main signal. An official method for identifying irradiated nuts will be proposed in 1992. Irradiation specific ESR-signals of the cellulose radical were not only found for nutshells but also for fresh fruit and some spices, while most of the irradiated spices and herbs could not be identified by ESR-measurements.


[The incidence of detecting antibodies to the hepatitis C virus in children in specialized children's institutions]. Marked differences in the distribution of markers of hepatitis C and B among children in closed children's institutions was established. The frequency of detection of anti-C100-3 in children varied from O in St. Petersburg to 29.4% in Nalchik. A high rate of infection in children is primarily associated with a low level of medico-sanitary service. The results indicate the advisability of specific prophylaxis of viral hepatitis C in specialized children's institutions.


[Gastric stump cancer with osteoplastic metastases and microangiopathic hemolytic anemia]. In a case of adenocarcinoma in the anastomosis of a stomach resected by the Billroth II technique osteoplastic bone infiltration and microangiopatic haemolytic anaemia are described as paraneoplastic syndromes. Both phenomena open up a great deal of differential diagnoses for consideration. Even when the extended bone infiltration could already be observed, the primary tumor macroscopically in the stomach was not visible. Even an unremarkable B II anastomosis should be biopsied as a matter of routine.


[The epidemic importance of the virological study of sewage for rotaviruses]. The results of correlation of rotavirus circulation indices among apparently normal and sick children and the rates of rotavirus antigen detection in sewage from an orphanage and an infectious ward of children's hospital are presented. It was established that there should be up to 1% of virus excretors for rotaviruses to be detectable in sewage of peripheral collectors of children's institutions. A significant increase in the rate of detection of rotaviruses in sewage may indicate an activation of the epidemic process of rotavirus infection.


Purification and characterization of Epstein-Barr virus gp340/220 produced by a bovine papillomavirus virus expression vector system. Our initial results with a bovine papilloma virus (BPV) vector expression system indicated that we could produce significant amounts of Epstein-Barr virus (EBV) gp340/220 in the supernatant of a mouse fibroblast cell line. We have now extended these findings to show that the truncated version of gp340/220, where the membrane anchor sequence is deleted, is produced even after extended passage of the cells, at a level of approximately 1 mg/4 x 10(8) cells. A simple purification protocol using Sephacryl S300HR and gelatin agarose gives a product which is greater than 90% pure. This product is recognized by anti-gp340 monoclonal antibodies from five different epitope groups and induces antibody that recognizes the authentic gp340/220 and neutralizes EBV in vitro. The purified gp340/220 can be used in ELISA and stimulates the proliferation of T-cell clones specific for gp340/220. These characteristics, together with the fact that BPV-transformed lines have been utilized for the production of pharmaceuticals for use in humans, suggest that this gp340/220 is suitable as a source of antigen for vaccination to prevent EBV infection and related diseases.


Biochemical and immunological characterization of a novel peptide carrier system using rotavirus VP6 particles. A system which allows for the efficient attachment of synthetic peptides to spherical virus-like particles assembled from the VP6 rotavirus nucleocapsid protein is described. This attachment was shown to be mediated by peptide-protein interactions and did not require additional chemicals for conjugation. The resulting large macromolecular structure was highly immunogenic for both the VP6 protein and the coupled peptides. The antibody response to peptides bound to VP6 particles was of higher titre and longer duration than that induced by other carriers. In addition, the response to VP6-coupled peptides was not affected by prior exposure to rotavirus and exhibited a range of immunoglobulin subclasses in the absence of an adjuvant. These data demonstrate that assembled VP6 spherical particles are useful carriers for low doses of synthetic peptides.


Evidence for retroviral capsid and nucleocapsid antigens in ovine pulmonary carcinoma. A retroviral etiology has been proposed for ovine pulmonary carcinoma (OPC); however, the putative virus (OPCV) has yet to be cultured. A Western immunoblotting assay using a panel of retroviral antisera was developed to further define the structural proteins of the virus associated with OPC and to confirm their presence in tumor samples of affected sheep. The results confirmed that the main structural viral component, the capsid protein (CA), was present in tumor materials (lung fluids, lavages, and tumor homogenates) but not in similar samples from control subjects. A second viral protein was detected in the tumor samples by antisera to the nucleocapsid protein (NC) of type D retroviruses. Both components could be purified from the tumor material in a manner consistent with association in viral particles. The cross-reactivity of the OPC antigens to other type B and D retroviruses was assessed. These results suggest that OPC antigens are closely related to the structural proteins of several type D primate retroviruses.


[Virus persistence in hepatitis A in monkeys]. A long-term complex observation of 16 cynomolgus monkeys (Macaca fascicularis) and 8 African green monkeys (Cercopithecus aethiops) with spontaneous and experimental hepatitis A revealed two forms of the illness: acute and chronic. Some monkeys developed undulating chronic course of the disease consisting of 2-6 waves. Others developed relapses (1 to 3) which occurred within 2-4 or 6-11.5 months of the infection. The morphological changes in the liver persisted for 7-28 months. Alaninaminotransferase elevations in the blood and HAV shedding in feces were observed periodically for 7-20 months. HAV persistence was documented by radioimmunoassay, enzyme immunoassay, immune electron microscopy and molecular hybridization. Persisting HAV was shown to remain pathogenic for monkeys. Virological evidence of the etiological association of HAV with chronic infection and late relapses has been obtained for the first time.


Host range analysis of a chimeric simian virus 40 genome containing the BKV capsid genes. Simian virus 40 (SV40) propagates poorly in cells from human embryonic kidney (HEK) and human fetal fibroblasts (HFF) while BK virus grows well in many human cell types. It has been suggested that sequences within the SV40 late region but not within the BKV late region may act to inhibit growth of virus in HEK and HFF cells. In order to test this and to identify a late region host range function, we have replaced the late region of wtSV40 DNA with the late region of RFV (a variant of BKV) to produce an intermolecular hybrid or chimera. The constructed SV40/RFV chimeric genome contained approx. 5900 base pairs, more than 650 base pairs greater than wtSV40. Nevertheless, when introduced by transfection the chimera appeared to be infectious. Three chimeric genomes were recovered from infected cells and all contained deletions of nearly 600 base pairs, exclusively at the region of the 3' terminal junction. Since all three chimeras propagated in human HFF and HEK cells, the RFV late region and not the RFV regulatory region possesses a host range function required for growth in human cells. Analysis of T-antigen gene expression suggests that the replacement of the SV40 late region with the BKV late region leads to full expression of the SV40 early region in human cells. Two chimeras exhibited a BKV-like host range and the third exhibited both a BKV and an SV40-like host range. We determined precisely which sequences were deleted in each chimera and we exchanged 3' terminal junction fragments containing these deletions, between two chimeras with different host ranges. From these experiments we demonstrated that: (1) The 3' terminus of the SV40 large T-antigen gene is required for growth of SV40/RFV in TC-7 and CV-1 simian cells but not for growth in human cells; (2) while the SV40 late region is refractory for growth in human cells, the RFV late region is not refractory for growth in simian cells; (3) the 3' terminus of the RFV T-antigen gene is not required for growth in human cells. The results of the 3' terminal junction exchanges and studies of early gene expression also demonstrate that BKV and SV40 can penetrate human and simian cells, even when they failed to grow in one cell type.


Determination of the coding capacity of the BamHI DNA fragment B of apathogenic Herpes simplex virus type 1 strain HFEM by DNA nucleotide sequence analysis. Herpes simplex virus type 1 (HSV-1) strain HFEM acquired an apathogenic phenotype due to a deletion within the DNA sequences of the BamHI DNA fragment B of the viral genome. In order to investigate the coding strategy of this particular region of the genome of HSV-1 strain HFEM the DNA nucleotide sequence of the BamHI DNA fragment B was determined. This analysis revealed that the BamHI DNA fragment B of HSV-1 strain HFEM comprises 6593 bp, corresponding to the nucleotide positions (np) 113322 to 117088 and np 120643 to 123465 of the genome of HSV-1 strain 17. According to these data the deletion of the genome of HSV-1 strain HFEM occurred between the np 117089 and 120642. The promoter region of the UL56 gene of HSV-1 strain HFEM is a part of the deleted DNA sequences. Therefore, this gene of HSV-1 strain HFEM is affected and cannot be expressed. The first 35 amino acid (AA) residues of the deduced amino acid sequence of the UL56 open reading frame (ORF) were found to be identical to the amino acid sequence of the UL56 genes of HSV-1 strains 17 and F. However, due to a deletion at np 3494 of the BamHI DNA fragment B of HSV-1 strain HFEM the amino acid composition of the predicted UL56 gene of HSV-1 strain HFEM is different from HSV-1 strain 17 between amino acid positions 36 and 233. In addition the deduced amino acid sequence of the IRL (inverted repeat of the long segment) copy of the IE110 gene of HSV-1 strain HFEM was found to be about 342 amino acids shorter than the amino acid sequence of IE110 gene of HSV-1 strain 17 (775 AA). This was based on a point mutation which was detected within the DNA sequences of Exon 3 of this copy of IE110 gene of HSV-1 strain HFEM.


Comparative analyses of the nucleocapsid genes of several strains of infectious bronchitis virus and other coronaviruses. The natural sequence variations of the nucleocapsid genes of the Gray, Arkansas99 (Ark99), and Holland52 (Holl52) strains of infectious bronchitis virus (IBV) were determined. These were compared with previously published sequencing data of other IBV strains, as well as other coronaviruses, in order to correlate the serological and evolutionary relationship of coronaviruses. IBV nucleotide sequence alignment shows that overall the sequences are highly conserved, with homologies from 91.1 to 96.5%. However, there are also two regions (730 to 800 and 1138 to 1166) that appear to be even more highly conserved. Overall, the nucleocapsid protein is highly variable both in size and composition between coronavirus major antigenic groups but is conserved within these groups. A phylogenetic tree of the nucleocapsid protein of various coronaviruses indicates that the coronaviruses fall into distinct groups that correspond to the three major antigenic groups; however, a phylogenetic tree of the IBV nucleocapsid shows that this does not hold true for the type specific antigenic groups of IBV.


PAb 2000 specifically recognizes the large T and small t proteins of JC virus. A monoclonal antibody, PAb 2000, has been derived which recognizes the large T protein of JC virus (JCV), but not the corresponding proteins of the related polyomaviruses BK virus (BKV) and SV40. The epitope bound by PAb 2000 was localized to the amino-terminal 81 amino acids of this multifunctional protein. As observed previously with several monoclonal antibodies that bind a similar region of SV40 large T antigen, PAb 2000 was found to interact with the small t antigen and the denatured form of large T antigen. This monoclonal antibody recognized a subpopulation of T protein, the abundance of which varied in different species of cells transformed by JCV. The availability of PAb 2000, the first JCV T antigen-specific monoclonal antibody, will facilitate the purification and biochemical characterization of the JCV oncoproteins.


Evolutionary analyses of five US bluetongue viruses using the cognate S2 genes. Full-length cDNA copies of S2 genes (segment 8), coding for non-structural protein 2 (NS2) of bluetongue virus serotypes 2, 11, 13 and 17 were selectively synthesized by a modified polymerase chain reaction (Clamp-R) and cloned into the PstI site of the pUC19 plasmid. Each of these S2 cognate genes was 1125 nucleotides in length with an initiation and a termination codon at nucleotides 20-22 and 1082-1084, respectively, resulting in a long open reading frame capable of coding for a protein of 354 amino acids. The deduced amino acid sequence of NS2 protein had a high concentration of lysine and contained a relatively low number of tryptophan and histidine residues. There was a highly conserved hydrophilic region at the carboxyl termini of predicted NS2 proteins in all five BTV serotypes, even though the amino acid sequence in this region in BTV-2 was more variable than in the other four serotypes. There was significant sequence homology of the cognate S2 genes at both the nucleotide and the amino acid levels. Phylogenetic analyses using the S2 gene sequences indicated that BTV-10, -11, -13 and -17 were more closely related and BTV-2 was the most distantly related serotype among the five US bluetongue viruses.


Typing of Hantaviruses from five continents by polymerase chain reaction. Hantavirus, a genus in the family Bunyaviridae, is comprised of at least four serologically distinct types: Hantaan, Seoul, Puumala and Prospect Hill. The present communication reports the use of polymerase chain reaction (PCR) for typing 27 independently isolated Hantaviruses from 5 different continents. Total cellular RNA was extracted from virus-infected Vero E6 cell monolayers by the acid guanidium thiocyanate-phenol-chloroform method. We have utilized 5 different sets of oligonucleotide primers ranging from 18 to 22 nucleotides in length; one set was specific for a conserved region of the S genomic segment and used as genus-specific primers, the other 4 sets of primers were designed from unique sequences of the M genomic segment such that each primer set was specific to only one serological type of Hantavirus. The PCR products were analyzed by restriction endonuclease digestion for further confirmation. We typed 10, 12, 3 and 1 isolates into Hantaan, Seoul, Puumala and Prospect Hill respectively. The results of PCR were 100% agreeable with that of serological typing, and thus, PCR can be used as an adjunct test with serological method(s) or an independent test for diagnosis and for typing of new isolates of Hantaviruses.


A cis-acting element within the hepatitis A virus 5'-non-coding region required for in vitro translation. Every picornavirus studied thus far has a sequence within the 5'-non-coding region that is required for internal ribosome binding and translation of the polyprotein. In an attempt to identify this region in hepatitis A virus we constructed a truncated hepatitis A virus (HAV) cDNA clone that contains the entire 736 bp 5' non-coding region (5'-NCR) and 754 base pairs of the viral capsid coding region (P1) under control of the SP6 promoter. In vitro transcription and translation of this transcript in a rabbit reticulocyte lysate yielded a protein product of about 29 kDa as analyzed by autoradiography following sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A series of mutations of this construct have defined a minimal sequence between bases 347 and 734 in the 5'-NCR that is required for efficient in vitro translation. The deleted constructs (D 523-734 and D 632-734) showed a reduced ability to translate in the rabbit reticulocyte lysate system in comparison with the full-length 5'-NCR construct, pH1489. The translation of these deleted constructs was artificially restored by the addition of a 5'-terminal methylated cap structure, m7GpppG, to the RNA. This increase in translational efficiency could be competed away with cap analog (m7GDP) thus indicating that this region is required for cap-independent internal ribosome binding for HAV translation.


[A neuropsychological analysis of the lack of school achievement among the pupils of mass schools]. We describe typology of neuropsychological organization underlying the cognitive difficulties of 6-10-years-old schoolchildren. In 10-20% subjects under study was revealed a deviant ontogenetic development with minimal cerebral disfunction, accompanied by systemic insufficiency of certain mental functions. Ontogenetic development of left-handed subjects as well as of right-handed ones with partial and/or family left-handedness was found to be atypical. About 80% of right-handed subjects with normal cognitive development displayed disorganization of some components of mental activity as a consequence of heterochronous morphofunctional maturation of the brain. Specific correction programs should be developed for each group of children under study.


[The functional role of enhanced cellular excitability and synaptic efficiency in the neocortex during learning]. In experiments with noncurarized and unanaesthetized rabbits was recorded pyramidal tract response in the course of conditioning of direct stimulations of the two points of the cortical surface. The data obtained point to temporary specificity in manifestation of the membrane and synaptic plasticity, to participation of these mechanisms in the processes proceeding in both cortical representations of paired stimuli, and to predominantly undirected changes of a degree of their involvement in both cortical areas. At the early stage of conditioning were demonstrated all the characteristics of the dominant state developing at this stage, and at the late one those of differential conditioning. A conclusion is drown that the reinforcement through the membrane plasticity leads to initial dominant increase of cellular excitability. On the background of the latter by means of summation mechanism the conditions are created for excitation transmission from the sensory link of a new bond to its motor output. Underlined by the mechanisms of synaptic plasticity gradual reorganization of the excitatory and inhibitory connections to the output elements of conditioned response determines and consolidates specialized character of the elaborated reaction.


[The mapping of brain biopotentials during the solving of a verbal task]. Reverse averaging of cortical potentials from the moment of the motor response followed the verbal task solving (anagram riddle) revealed some brain potentials correlations with the process of a decision making. In the case of task solving the negative frontal wave with the latency 900-400 ms from the motor response was recorded. Intracortical interaction mapping of this potential showed the regular patterns of cortical functional connections in different frequency ranges (alpha, beta). Successful solving of the task was characterized with predominant interaction foci topography in the frontal and left-temporal cortical areas in alpha band and parietal zones in beta. The absence of the task solution was characterized with the parieto-occipital interaction foci in alpha band and their frontal localization in beta.


[An EEG analysis of the structural interrelationships of the thalamofrontal system in the recovery of conditioned-reflex behavior in amygdalectomized rats]. Unilateral damage of the baso-lateral amygdala in white non-strain rats led to decrease of the number of instrumental responses and increase of their latencies in the course of the first two weeks after the surgery. This period was characterized by increase of the delta- and theta-frequency bands in electrograms of the orbito-frontal cortex and dorsomedial nucleus of the thalamus. During the next two weeks this increase was observed only in the orbito-frontal cortex. Functional reorganization of interconnections between the limbic and thalamo-cortical brain structures is supposed to underly phenomena observed.


[The role of the genotype in the variability of human evoked potentials at different stages of ontogeny]. The determinants of individual differences in visual EPs to stimuli of different type depending on the age of subjects were studied. EPs to flash, checkerboard pattern and some other stimuli with semantic aspect were recorded in three groups of MZ, DZ twins aged 8-9, 10-12, 18-25 years. The heritability of EPs parameters is not ontogenetically stable characteristic. It changes from one age group to another in different way for separate EPs components and parameters depending on their nature, type of stimulus, area of recording. Most of all genotypical influences are manifested in EPs parameters of 10-12 years old subjects, especially the middle latency components from vertex area and latent periods of EPs in comparison with their amplitudes.


[A comparison of the self-stimulation reaction and of conditioned place preference with fenamine administration in rats]. The reinforcing properties of different doses of amphetamine (1 and 5 mg/kg) were examined using two variants of self-stimulation reaction (in the Skinner box and locomotor self-stimulation in a shuttle box) and place preference test. Amphetamine in dose of 1 mg/kg increased the frequency of self-stimulation in the Skinner box and prolonged the time of rat staying in active zone of a shuttle box to a greater degree than 5 mg/kg of the drug. On the contrary, the aversive phase of self-stimulation, determined by a coefficient of "disagreement", grew higher after 5 mg/kg amphetamine than following 1 mg/kg. The study of effects by place preference test revealed the other regularity: the most positive reinforcing properties possessed the drug in a dose of 5 mg/kg. Thus, there are dissociation between the two doses of amphetamine (1 and 5 mg/kg) in their action on different physiological conditioned responses. The mechanisms of this dissociation are discussed.


[The dependence of the neuronal reactions of the sensorimotor cortex to a simultaneous complex stimulus on the level of the differentiation of its components]. On alert animals the change was studied of the neuronal activity of the sensorimotor cortical area of cats brain in dependence on the level of differentiation of the components of simultaneous heteromodal complex stimulus. According to the character of this dependence and a number of other parameters two groups of neurones were singled out in the sensorimotor cortex. It was shown that parameters of reactions of all recorded neurones of the sensorimotor cortex to the positive conditioned signal were the first established after consolidation of the animal conditioned motor activity. In the course of elaboration following parameters changed: expressiveness, intensity, duration and value of latency. Reactions of neurones of both groups to the inhibitory stimuli were stabilized only after consolidation of the habit of differentiation. Responses of the first group neurones changed only by the pattern of discharge, while the responses of the second group neurones could change by expressiveness of response, its sign, duration and value of latency. Oscillations of the differentiation level after finishing of the elaboration of inhibitory conditioned reactions affected only the responses of the second group neurones to complex components.


[The role of visual stimuli in the acquisition of place preference with morphine reinforcement]. In experiments with rats was studied the role of visual stimuli in conditioning of place preference in a T-shaped chamber, certain stimuli being combined with morphine injection (5 mg/kg). Intensity of illumination of the chamber sections, colour of walls, and the orientation of dark stripes on the walls were varied. Testing the animals' behaviour began with determining the reward properties of certain visual stimuli on the basis of the total time of the animals' stay in a chamber. Procedure of combining the visual stimuli with morphine was repeated four times. Associative properties of the stimuli were revealed by comparing the learning efficiency in different experimental groups. Learning within the framework of the place preference model was concluded to depend upon the reward properties of the visual stimuli, whereas the behaviour itself to be based on the mechanism of the instrumental but not the classical conditioned response.


[The neuronal reactivity and interaction of the rat cerebral cortex during the microiontophoretic action of acetylcholine in a model negative learning situation]. Unit activity of 46 pairs of neurons of sensorimotor cortex of rats was studied in a model situation of habituation to repetitive microiontophoretic applications of acetylcholine (ACh). The difference between the types of reactions to ACh of two neighbouring neurons recorded by the same microelectrode was observed on 37% of cases. The difference between the dynamics of activity of neurons with similar patterns of reactions during repetitive action of stimuli was also shown. Stability of the excitatory connections between two neighbouring neurons under the conditions when one of them demonstrated the habituation to repetitive action of ACh were indicated by analysis of cross-correlation histograms.


[The morphofunctional characteristics of the neurons in the sensorimotor cortex of old rabbits during the trace assimilation of rhythm]. Extracellular neuronal activity was recorded from 460 neurons from alert young (5-7 months), middle-aged (54-65 months) and old (66-85 months) rabbits. Trace rhythmic activity of sensorimotor cortical neurons was examined after long-lasting (10-20 min) rhythmic (0.5-2 Hz) electrocutaneous stimulation of the contralateral forelimb. Spectral analysis of spike activity showed age-related differences in capability of producing a rhythm of previous stimulation in spontaneous neuronal activity. In young animals propriate rhythmic fluctuations of firing rate appeared after the first or second sessions of stimulations (on the first experimental day), in middle-aged ones--after 2-4 sessions (on the second or third days); cortical neurons in old rabbits did not exhibit trace rhythmic activity. Significant morphological changes in glial and neuronal cells were observed in sensorimotor cortex of old rabbits. It is proposed that morphological deteriorations may be the reason of the impairement of trace processes during aging.


[The reactivity of the hypophyseal-adrenal system in rats with different forms of adaptive behavioral pathology]. The dexamethasone suppression test (DST) was applied to male Wistar rats with different models of depression: group with the learned helplessness, group with informational neurosis provided by time-deficit conditioned avoidance training, as well as groups of rats of two strains selected for low (KLA)--and high (KHA) avoidance learning. The pre-dexamethasone basal and stress-induced corticosterone levels were similar in intact rats and those exposed to inescapable shock. The dexamethasone administration (5 mkg/kg) failed to decrease the serum corticosterone level in rats with learned helplessness. The informational neurosis increased significantly the basal corticosterone level and decreased the stress response. Serum corticosterone levels were similar in KLA and KHA rats. These results give evidence that two stress-induced rat models of depression with similar behavioural disturbances (reduction of escape/avoidance reactions) exhibit marked differences in the activity of hypothalamo-pituitary-adrenal (HPA) axis.


[The open-field behavior of neuroticized rats]. A detailed study has been carried out of the behaviour in the open field of the lines of rats with different functional state of the nervous system after prolonged neurotization in dependence on the phase of circadian rhythm and terms after the end of the influence. Significant changes have been shown by the amplitude of circadian rhythms of emotionality and motor activity. It is established that the reaction of the rats lines to prolonged neuroticizing action depends on the phase of the circadian rhythm, the time after its termination and on genetically determined level of excitability of the nervous system.


[The effect of the removal of the epiphysis on the antidepressive properties of paradoxical sleep deprivation in rats]. After REM sleep deprivation antidepressant shifts in forced swimming with increase of time of immobility and decrease of rhythmical index of depression were observed. Pinealectomy did not remove, but attenuated these behavioural changes.