Improved diagnosis of chronic hepatitis C virus infection by detection of antibody to multiple epitopes: confirmation by antibody to synthetic oligopeptides. Serum samples from 226 patients covering a wide spectrum of liver disease were tested for antibodies to hepatitis C virus (HCV) using both first and second generation enzyme linked immunosorbent assays. Selected sera were also tested by peptide immunoassays, by the four-antigen recombinant immunoblot assay (RIBA II), and for viral genome by the polymerase chain reaction. Antibody to c100-3 was detected in 61% of patients with chronic non-A, non-B (NANB) hepatitis and/or 46.5% with presumed NANB-related cirrhosis by the first generation test. These figures increased to 77% and 58% when antibodies to recombinant structural and non-structural HCV antigens were sought by the second generation assay. Supplemental testing against peptide Sp75 and Sp65/sp67 confirmed that reactivity of sera by second generation assays was due to antibodies to the additional structural and non-structural antigens. Samples negative by the first generation assay were not confirmed by the supplemental assay using peptides Sp75 and Sp65/Sp67. HCV RNA was detected in 60% of the anti-HCV positive sera tested, most of which were also RIBA II positive. Our findings confirm that the introduction of the structural and non-structural antigens, especially the putative nucleocapsid protein, improves sensitivity of detection of antibodies to HCV, and facilitates diagnosis in patients with "cryptogenic" chronic hepatitis.


Virus-like particles with reverse transcriptase activity associated with Kawasaki disease. To investigate the etiologic agent associated with Kawasaki disease (KD), we initially established a cocultivation system using concanavalin A (Con A)-stimulated lymphoblastoid cells obtained from each retrovirus-seronegative individual's peripheral blood mononuclear cells (MNCs) cocultivated with each of 1) 40 patients with KD, 2) 10 patients with other viral infection and skin rash, and 3) 10 age- and sex-matched normal controls. Five major findings suggested that virus-like particles with reverse transcriptase (RT) activity are associated with KD. First, RT activity appeared significantly higher on day 12 after the onset of fever in the KD patients than in those with other viral infections and normal controls (dTMP incorporation: 3,645 +/- 248 vs. 434 +/- 50 vs. 412 +/- 46 cpm, P < 0.0001). Second, the RT activity was not endogenous, because the Con A-stimulated lymphoblastoid cells were obtained from the individuals who were negative for retrovirus. Third, virus-like particles (80-100 nm in diameter) by electron microscopy were found in the concentrated pool supernatants of particulate fraction containing RT activity subjected to sucrose density gradient, obtained from KD patients. Fourth, the viral product, a 31.4 kilodalton molecule, was detected by SDS-PAGE after internal labelling (methionine-S35) and density gradient centrifugation. Fifth, using a "retrovirus universal pol gene region" as a primer and the RT-PCR method, a retrovirus-specific band was detected in the cocultivated supernatants obtained from four KD and one AIDS patients but not in patients with rubella or in healthy controls.(ABSTRACT TRUNCATED AT 250 WORDS)


Serum hepatitis C virus (HCV)-RNA and response to alpha-interferon in anti-HCV positive chronic hepatitis. Hepatitis C virus (HCV) replication was assessed before and during alpha-interferon (IFN) treatment in 22 anti-HCV positive patients with posttransfusion or sporadic chronic hepatitis (CH). Eleven patients were "responders" and 11 patients "non-responders" to IFN. Thirteen anti-HCV negative healthy subjects and five anti-HCV negative patients with autoimmune CH served as controls. Serum HCV-RNA was detected by the polymerase chain reaction (PCR) in all untreated anti-HCV positive patients but in none of the anti-HCV negative subjects. PCR primers from the 5'-noncoding (NC) region were more sensitive than primers from a non-structural (NS5) region in detecting HCV-RNA (21/22, 95% vs. 7/22, 32%, respectively). Positive strand HCV-RNA titre and positivity rate for the negative strand were similar in responders and non-responders before IFN treatment, as well as anti-c100-3 titre by enzyme-linked immunosorbent assay (ELISA), and anti-5-1-1, anti-c33c, anti-c22 positivity rate by immunoblot assay (RIBA). HCV-RNA positivity by both NC and NS primers was more frequent before IFN among responders. During IFN treatment, serum HCV-RNA was detectable, mostly at low titres, in 1 (NC positive) of the 11 responders and in 9 (4 NS positive and 5 NC positive) of the 11 non-responders. Among the four non-responders who were NS positive during IFN, three were NC positive before IFN. Serum HCV-RNA was always found in our post-transfusion or sporadic anti-HCV positive patients with CH. Viraemia generally decreased during IFN treatment, but no available HCV markers clearly distinguished responders from non-responders before IFN treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of S-100 protein, epithelial membrane antigen, carcinoembryonic antigen and alpha fetoprotein in normal salivary glands and primary salivary gland tumors. The distribution of S-100 protein, epithelial membrane antigen, carcinoembryonic antigen and alpha fetoprotein was studied in 38 primary salivary gland tumors. S-100 protein, a useful marker of myoepithelial cells, was demonstrated in some benign tumors. Carcinoembryonic antigen expression was consistently positive in adenoid cystic carcinoma. Demonstration of epithelial membrane antigen helped to confirm the epithelial nature of some neoplastic cells. Alpha fetoprotein was not expressed in any of the cases examined. No correlation was found between immunopositivity and tumor behavior in the present series.


Electrophoretic detection of salivary alpha-amylase activity. Fresh samples of human whole saliva containing approximately 20-40 micrograms protein were analyzed using SDS-polyacrylamide slab gel electrophoresis systems. More than 20 protein bands were revealed by Coomassie Brilliant Blue R 250 staining. Some of the protein bands were shown to be glycoprotein-positive with PAS (periodic acid-Schiff) reagent. The protein bands with alpha-Amylase activity appeared within a molecular weight range of 120,000-180,000, which is 2 to 2.8 times higher than the normal molecular weight reported for alpha-Amylase from parotid saliva, and showed positive staining with PAS reagent. These results show that the alpha-Amylase in whole saliva appears to exist in a macromolecular form which is not dissociated in the presence of sodium dodecyl sulfate (SDS).


Insecticide poisoning in pregnancy. A case report. A 25-year-old multigravida drank an organochlorine insecticide in an attempt to commit suicide at 16 weeks of pregnancy. This resulted in the death of twin fetuses and vaginal bleeding. The pregnancy was terminated using gemeprost, syntocinon and later surgical evacuation of the uterus. She developed respiratory arrest after the fetuses were aborted and also after surgical evacuation. On both occasions, she was intubated and mechanically ventilated. We believe that organochlorine insecticides may be fetocidal. In addition, surges in blood levels may occur during termination of pregnancy, delivery or evacuation of retained products of conception.


[Assessment of air pollution health effects on respiratory organs]. During the past three decades, industrial expansion in Japan has been remarkable, resulting in a numerous number of chemical substances got synthesized. Unfortunately, our living environment has concomitantly been polluted with such substances released through industrial activities and our daily lives, and health injuries have occasionally occurred in the human population. Although the critical conditions in the 1960s were overcome by the countermeasures we took, the potential for environmental pollution still remains. In the present social and economical situation, the management of environmental pollutants should be decided depending upon more quantitative and predictive evaluation of their health effects. In this paper the author tries to evaluate the relation of air pollution to chronic bronchitis, bronchial asthma and pulmonary cancer from a quantitative point of view.


Role of cyclic AMP in prostaglandin-induced modulation of acetylcholine release from the myenteric plexus of guinea pig ileum. Prostaglandins (PGs) have modulatory effects on spontaneous and nicotine-induced release of acetylcholine (ACh) from the myenteric plexus of guinea pig ileum. To determine whether cyclic AMP is involved in the mechanisms of these effects, we studied ACh release under conditions that inhibit PG synthesis. Indomethacin (IND), a cyclooxygenase inhibitor, inhibited ACh release concentration-dependently. The effect of the maximally inhibitory concentration of IND (2.8 microM) on nicotine-induced ACh release were reversed concentration-dependently by PGE2, forskolin, 3-isobutyl-1-methylxanthine (IBMX) and 8-bromo cyclic AMP. These compounds caused concentration-dependent reversal of the inhibition of spontaneous ACh release by IND, but their concentrations for restoration of spontaneous release were higher than those for restoration of nicotine-induced release. The effects of PGE2 and forskolin or IBMX were not additive in reversing the inhibition of nicotine-induced ACh release by IND. Neither forskolin nor 8-bromo cyclic AMP alone had any significant effect on either release. These results showed that increase in the level of cyclic AMP in myenteric cholinergic neurons restored ACh release from the tissue whose PG level had been lowered by IND and indicated that endogenous PGs may modulate the level of intraneuronal cyclic AMP.


[Chronic obstructive lung disease: does accompanying arrhythmia have any clinical significance?]. Clinical significance of cardiac arrhythmias in patients with advanced and stable chronic obstructive pulmonary disease was assessed 22-24 hour Holter monitoring revealed supraventricular and ventricular arrhythmias in all 65 patients. There was a great variation of arrhythmia quality and quantity in the study group. Among others, 1808 complex ventricular extrasystoles in 33 patients and 302 episodes of nonsustained ventricular tachycardia in 19 patients were recorded. No sustained symptomatic ventricular arrhythmias were observed. We found no correlation between the intensity of ventricular and supraventricular extrasystoles and clinical parameters as assessed by echocardiography, spirometry and gasometry. However, patients with complex ventricular arrhythmias had larger right ventricular diameter in echocardiography. Of 65 studied patients, 14 (21%) died during follow-up ranging from 1 month to 3.2 years, mean 12.2 months. Two outpatients died suddenly. These 14 patients had larger right ventricle and left atrial dimension, more pronounced PaO2 decrease as well as lowered FEV1 as compared to the survivors. No differences in the incidence of cardiac arrhythmias were noted. Summarizing, despite the great prevalence of cardiac arrhythmias in patients with advanced stable COPD we found no life-threatening ventricular arrhythmias during approximately 1500 hours of holter monitoring. Cardiac arrhythmias seem not to influence the prognosis in these patients.


Introduction: the metabolic cardiovascular syndrome. During the last few years new knowledge concerning risk factors for cardiovascular diseases (CVD) has emerged. Many of these are metabolically interrelated. The clustering of a variety of metabolic disorders in individuals prone to develop CVD has led to the launching of the so-called metabolic cardiovascular syndrome (MCVS). At the XIIIth Scandinavian Congress of Cardiology in Oslo, Norway, on June 1991, a satellite symposium was arranged on the MCVS. The present supplement comprises articles written by the symposium speakers, who are all experts in various fields of relevance to the MCVS. In addition, Per Björntorp, Stevo Julius, and Arne Westheim were invited to contribute articles to make the coverage of the topic more complete. The present introduction summarizes some of the clinical and biochemical features characteristic of the MCVS, with a special reference to the multiplicity of biochemical changes. The possibilities for at least three basic disorders being of primary or causal importance in different individuals are stressed. These are chronic sympathetic overactivity, insulin resistance, and central or visceral obesity. By including the related changes in the hemostatic systems with "hypercoagulability" and "hypofibrinolysis," whereby the importance of thrombosis for the clinically often dramatic complications of the MCVS is stressed, the more specific name atherothrombogenic syndrome (ATS) has been proposed. The non-pharmacological treatment of the syndrome with low-caloric diet and physical exercise is emphasized.(ABSTRACT TRUNCATED AT 250 WORDS)


Carbohydrate metabolism, insulin resistance, and metabolic cardiovascular syndrome. Insulin resistance is seen in several pathophysiological conditions, such as obesity, diabetes mellitus, and essential hypertension. This means that a greater than normal amount of insulin is needed to give a normal biological response. A major biochemical defect in insulin resistance seems to be a defect in the intracellular nonoxidative metabolism of glucose in muscle cells. However, in many individuals, there is also increased hepatic glucose output. The result of insulin resistance in individuals with normal insulin-secreting capacity is hyperinsulinemia, a potential risk factor for cardiovascular disease.


Hypertension and the metabolic cardiovascular syndrome: special reference to premenopausal women. It has been proposed that hyperinsulinemia may not constitute a cardiovascular risk in women, and that the metabolic risk profile is less apparent in women than in men. In two different studies, we have assessed the interrelationship between classical coronary risk factors in women with untreated essential hypertension and looked for possible hypertensive-normotensive differences. Hypertensive women (HT1, 156 +/- 2/98 +/- 1 mm Hg, n = 18) in study I turned out to be overweight and had nearly three times higher fasting serum insulin levels than the normotensive control subjects (NT1, 118 +/- 3/77 +/- 2 mm Hg, n = 9). HT1 women with a body mass index (BMI) above 25 kg/m2 had significant higher insulin levels than HT1 women with a BMI less than 25 kg/m2, and when adjusting for BMI the hypertensive-normotensive difference in insulin levels was lost. In HT1 women, the serum insulin level correlated positively to the BMI and triglycerides. In study II, insulin was positively associated with the systolic blood pressure in HTII women (150 +/- 3/99 +/- 1 mm Hg, n = 29), and a negative correlation appeared between the glucose/insulin ratio and the systolic as well as diastolic blood pressure. No difference was observed in BMI and insulin between HTII and NTII women (121 +/- 3/79 +/- 1 mm Hg, n = 18). In HTII women, plasminogen activator inhibitor (PAI-1) levels were higher and the euglobulin clot lysis time prolonged compared to NTII women. PAI-1 was positively correlated to insulin and triglycerides and negatively to high-density lipoprotein (HDL) cholesterol in HTII women. Strong associations between potential cardiovascular risk factors seem to be present even in untreated women with mild hypertension, with insulin being correlated to hypertension, BMI, fibrinolytic activity, triglycerides, and HDL cholesterol.


Lipoproteins, lipases, and the metabolic cardiovascular syndrome. High levels of plasma triglycerides and very-low-density lipoproteins and low levels of high-density lipoproteins are consistently found in the metabolic cardiovascular syndrome. These changes are exaggerated postprandially. In the liver, synthesis and secretion of triglyceride-rich particles are increased. In addition, the removal capacity of plasma triglycerides is decreased, due to downregulation of lipoprotein lipase in skeletal muscles by hyperinsulinemia. Resistance to insulin-stimulated glucose uptake is believed to be the main pathogenetic factor. However, increased flux of free fatty acids from abdominally localized adipose tissue must also be considered when discussing pathogenesis. Treatment is primarily nonpharmacological, with diet and increased physical activity.


Abdominal fat distribution and the metabolic syndrome. Recent studies have indicated that the waist/hip circumference ratio (WHR), an index of abdominal fat distribution, is a risk factor for cardiovascular disease and diabetes, in parallel with other previously established risk factors. Obesity, without taking fat distribution into account, seems to be associated with WHR in its relationship to the metabolic risk factors for these diseases. The important component of the WHR is probably the mass of visceral fat. This cluster of phenomena constitute what has recently been called the metabolic syndrome or syndrome X. Visceral fat mass is probably increased by a multiple endocrine aberration, where steroid hormones are important. This seems to cause insulin resistance by direct effects on the periphery, which may be amplified by the metabolism of the enlarged visceral adipose tissues.


Hypercoagulability and reduced fibrinolysis in hyperlipidemia: relationship to the metabolic cardiovascular syndrome. Hypercholesterolemia is seen as an important risk factor for coronary artery disease (CAD), as the incidence of CAD is strongly correlated with the level of serum cholesterol in epidemiological studies. However, hypercoagulability and reduced fibrinolytic capacity, often seen in survivors of myocardial infarction, are associated with hypertriglyceridemia (possibly concomitant with low levels of high-density lipoprotein cholesterol) and not with increased levels of total or low-density lipoprotein cholesterol. The important role of thrombogenesis in CAD is supported by the fact that initial high levels of plasma fibrinogen, coagulation factor VII (VIIc), and plasminogen activator inhibitor (PAI-1) are all independent risk factors for CAD or recurrent myocardial infarction as found in multivariate analyses of epidemiological studies. Furthermore, high plasma levels of VIIc and PAI-1 are associated with hypertriglyceridemia, reduced glucose tolerance, overweight, and hyperinsulinemia. The contribution of thrombogenic risk factors to the metabolic cardiovascular syndrome (MCVS) is thus established. Diet intervention is preferable for the normalization of hypercoagulability and hypofibrinolysis associated with MCVS. In familial combined hyperlipidemia, however, and especially with concomitant thromboembolic disease, diet alone is often not sufficient, and drug treatment with anticoagulants and/or lipid-lowering drugs may be necessary.


The sympathetic nervous system may modulate the metabolic cardiovascular syndrome in essential hypertension. The association between blood pressure and coronary artery disease may be caused by a concurrence of atherogenic biochemical abnormalities in hypertensive patients, i.e., the metabolic cardiovascular syndrome (increased total cholesterol, triglycerides, and insulin; decreased high-density lipoprotein (HDL) cholesterol; and insulin resistance, glucose intolerance, and blood platelet dysfunction). There are numerous reports of sympathetic nervous system overactivity in hypertensive subjects that could be of importance for the pathophysiology of the high blood pressure. Plasma catecholamines have metabolic hormonal effects at concentrations slightly above low normal resting levels. Even transiently and certainly chronically raised plasma catecholamine levels may cause biochemical abnormalities. Catecholamines may raise total cholesterol, triglycerides, and insulin, decrease HDL cholesterol, and cause insulin resistance and glucose intolerance, and recent evidence supports an in vivo influence of epinephrine on blood platelets, causing dysfunction in hypertensive subjects. Thus, the sympathetic nervous system may modulate the metabolic cardiovascular syndrome in essential hypertension. Hypertensive subjects may respond to environmental stimuli with larger sympathoadrenal responses than normal subjects. Furthermore, emotional stress has been associated with coronary artery disease. Thus, the metabolic hormonal effects of catecholamines, by causing the metabolic cardiovascular syndrome, may be the crucial link between "stress" and cardiovascular disease.


Early association of sympathetic overactivity, hypertension, insulin resistance, and coronary risk. Whereas this review is based on the available evidence, interpretation of the data is entirely hypothetical and subjective. To a large degree the review reflects our long-standing fascination with the role of the sympathetic overactivity in hypertension. The basic tenet is that sympathetic overactivity in hypertension may lead to a number of secondary changes, all of which are conductive to coronary heart disease. We also propose that insulin resistance and hypertension are associated through an underlying hemodynamic abnormality and again find good evidence for a possible role of sympathetics in the genesis of such hemodynamic changes. Future research in our laboratory will be oriented toward experimental testing of various aspects of the proposed sympathetic overactivity/hypertension/insulin resistance hypothesis.


Physical activity and the metabolic cardiovascular syndrome. Hyperinsulinemia, lipid abnormalities, and impaired fibrinolytic capacity may accompany hypertension, and comprise the metabolic cardiovascular syndrome. They are all independent risk factors for coronary artery disease. Lifestyle modifications can reduce the coronary artery risk. Physical activity has been shown to improve insulin sensitivity, reduce hypertriglyceridemia, improve fibrinolytic capacity, and reduce the blood pressure. A sedentary lifestyle is associated with increased mortality compared to a physically active lifestyle. Hypertensive patients, who often have other atherothrombogenic risk factors, should be encouraged to undertake physical activity as an adjuvant to other nonpharmacological interventions as well as to pharmacological treatment.


The metabolic cardiovascular syndrome: syndrome X, Reaven's syndrome, insulin resistance syndrome, atherothrombogenic syndrome. A short review of the metabolic cardiovascular risk syndrome (MCVS) is given. Traditionally, cardiovascular risk has been associated with three so-called "main" risk factors; hypercholesterolemia, hypertension, and smoking. In addition, the association between diabetes and cardiovascular disease has been known for many years in clinical medicine. Primarily, these risk factors have been regarded separately as independent factors, although epidemiological studies showed intercorrelations between them. However, it is now well accepted that relatively few at-risk individuals have only one risk factor, and in many cases a whole "symphony" of factors play together to create what we might call an individuals' risk profile. As an example, very often essential hypertension has been regarded as a disease in itself, which can be successfully treated just by lowering the blood pressure by drugs. When such a strategy obviously failed, the association of elevated blood pressure with dyslipoproteinemia and impaired glucose tolerance attracted more attention, particularly when it was realized that many antihypertensive drugs affected risk in MCVS in a possible negative way. The most important etiologic factor of MCVS is (besides genetics) an excessive caloric intake compared to what the individual spends in physical activity. In the clinical setting, the most important findings of MCVS are central obesity, dyslipoproteinemia with low high-density lipoprotein (HDL) cholesterol, hypertension, reduced insulin sensitivity in peripheral tissues, and increased thrombogenicity. The reduced insulin sensitivity leads to a compensatory increase in beta-cell insulin production, and thereby hyperinsulinemia.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of intracerebral iloprost injections on motor activity and chemically-induced seizures in rats. The effects of iloprost, a chemically stable analog of prostacyclin, on motor activity, pentylenetetrazol (PTZ)- and strychnine (ST)-induced seizures were studied in rats. Depression on motor activity was observed after a 500 ng/icv dose. Thus, both spontaneous locomotor activity and exploratory behavior were significantly reduced. While iloprost was ineffective against ST-induced seizures, it produced dose-dependent inhibition of PTZ-induced seizures. ED50 (95% confidence limits) value of iloprost for the suppression of clonic convulsions induced by PTZ was 224.96 (100.43-504.00) ng/icv. Anticonvulsive effect of iloprost was significantly potentiated by clonazepam pretreatment. In this case ED50 of iloprost was 39.40 (23.88-65.01) ng/icv. Unilateral iloprost injections into substantia nigra pars reticulata in a relatively lower dose range (0.5-2.0 ng/ic) also dose-dependently inhibited PTZ-induced seizures. In comparison to other prostanoids iloprost seems to have more potent and selective anticonvulsive activity against PTZ-induced seizures without marked motor depressant action in rats. It is further suggested that antiseizure effect of iloprost might be mediated by GABAergic inhibitory mechanisms.


Identification of Oklahoma isolate as a strain of Pseudomonas pseudomallei. Based on the morphological, physiological, biochemical and nutritional characteristics, cellular fatty acid and lipid composition, ubiquinone-8 as the major respiratory quinone, guanine-plus-cytosine content of DNA, DNA-DNA homology value, and sequence alignment of 16S rRNA nucleotides, Oklahoma isolate was reidentified as a strain of Pseudomonas pseudomallei.


Proposal of Burkholderia gen. nov. and transfer of seven species of the genus Pseudomonas homology group II to the new genus, with the type species Burkholderia cepacia (Palleroni and Holmes 1981) comb. nov. Based on the 16S rRNA sequences, DNA-DNA homology values, cellular lipid and fatty acid composition, and phenotypic characteristics, a new genus Burkholderia is proposed for the RNA homology group II of genus Pseudomonas. Seven species in this group were transferred to the new genus. Thus seven new combinations, Burkholderia cepacia (Palleroni and Holmes 1981), Burkholderia mallei (Zopf 1885), Burkholderia pseudomallei (Whitmore 1913), Burkholderia caryophylli (Burkholder 1942), Burkholderia gladioli (Severini 1913), Burkholderia pickettii (Ralston et al 1973) and Burkholderia solanacearum (Smith 1896) were proposed.


[Enzyme immunoassay stability of alpha tumor necrosis factor in plasma and serum]. With an enzyme immunoassay (ELISA) the stability of tumor necrosis factor alpha (TNF-alpha) has been determined in serum and plasma with and without addition of a protease inhibitor (Aprotinin, Trasylol) 24 hours, as well as seven days after blood collection. It has been observed, that after blood collection the concentration of TNF-alpha decreases first and increases again six hours, (+4 degrees C) respectively twelve hours (-20 degrees C) later. Reasons for that might be: The active form of TNF-alpha is a trimer, the presence of TNF-binding proteins, the presence of TNF-antibodies. Furthermore it has been found, that TNF-alpha has the highest stability in plasma at -20 degrees C and -70 degrees C. The addition of Aprotinin had no significant influence on the stability.


[The cellular and lysosomal fragility of pancreatic acinar cells after ligation of pancreatico-biliary duct in the rat and the protective effects of nafamostat mesilate]. To explore the changes of exorine pancreas in the early stage after ligation of pancreatico-biliary duct (PBD) in rat, we evaluated the changes of serum amylase levels, pancreatic water content as well as the changes of subcellular distribution of cathepsin B in the acinar cells. We also evaluated the lactic dehydrogenase (LDH) discharge from dispersed acini as an index of cellular fragility and cathepsin B leakage from lysosomes as an index lysosomal fragility in vitro study as well as the protective effects of a new synthetic protease inhibitor, nafamostat mesilate (FUT 175) in this model. After ligation of PBD, the serum amylase levels and pancreatic water content, increased significantly compared with those of the control group, but returning to the normal levels at 18 hours. The redistribution of cathepsin B in acinar cells was found and both the cellular and lysosomal fragility increased significantly compared with those of the control groups, shown their peak changes at 12 hours after ligation of PBD. But continuous intravenous administration of FUT 175 (2mg/mg.hr) remarkably attenuated all the parameters. These results indicate the important roles of lysosomal enzyme and lysosomal fragility in the pathogenesis of acute pancreatic injuries in this modes, and the clinical usefulness of FUT 175 in the treatment acute pancreatitis.


G-protein mediating the slow depolarization induced by FMRFamide in the ganglion cells of Aplysia. Application of FMRFamide (Phe-Met-Arg-Phe-NH2) induced a slow depolarization in neurons of the Aplysia abdominal ganglion. In voltage-clamped cells, FMRFamide induced a slow inward current that increased when the membrane was depolarized beyond -85 mV, showing a negative slope conductance. However, this inward current never reversed to outward current when hyperpolarized beyond the equilibrium potential for K+. The FMRFamide-induced response was markedly augmented in Ca(2+)-free media, but depressed in Na(+)-free media. It was unaffected by a change in external potassium. Intracellular injection of guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) significantly depressed the FMRFamide response in a dose-dependent way. Injection of cholera toxin (CTX) which did not cause any current response, selectively and irreversibly blocked the FMRFamide response. Neither 3'-deoxyadenosine, an inhibitor of adenylate cyclase, nor H-8, an inhibitor of cyclic adenosine 3',5'-monophosphate (cyclic AMP)-dependent kinase, depressed the FMRFamide response. 3-Isobutyl-1-methylxanthine (IBMX) did not augment the FMRFamide response appreciably. The FMRFamide response was not occluded at all by a relatively large injection of 8-bromo-cyclic AMP. It was concluded that the FMRFamide response is produced by the opening of the voltage-dependent Na(+)-channels via activation of a certain CTX-sensitive G-protein which is different from conventional "Gs" that activates adenylate cyclase.


Enzymatic inactivation of tachykinin neurotransmitters in the isolated spinal cord of the newborn rat. A mixture of peptidase inhibitors increased the magnitude of the saphenous nerve-evoked slow depolarization of a lumbar ventral root and prolonged the similarly evoked inhibition of monosynaptic reflex (MSR) in the isolated spinal cord of the newborn rat in the presence of naloxone. The saphenous nerve-evoked MSR inhibition was curtailed by a tachykinin antagonist, GR71251, and after the treatment with GR71251, the peptidase inhibitor mixture no more prolonged the MSR inhibition. The present results suggest that enzymatic degradation plays a role in the termination of action of tachykinins released from primary afferents in the newborn rat spinal cord. The results provide a further support for the notion that tachykinins serve as neurotransmitters in the spinal cord of the newborn rat.


Synovial sarcoma: a new approach to the histological classification and a comparison with malignant mesothelioma. Immunohistochemical study with anti-mesothelial cell serum. Routine paraffin-embedded sections from synovial sarcomas in 70 patients were stained immunohistochemically with monoclonal anti-cytokeratin antibody and serum against mesothelial cells. Positive reactions occurred in the epithelioid cells of biphasic tumors only. On the ground of the reaction with anti-mesothelial serum a hypothetical scheme of cell differentiation in synovial sarcoma was set up.


Aspiration biopsy of the breast tumors. Analysis of diagnostically difficult cases. In this paper the authors described those of the breast fine needle aspirates which were difficult for interpretation e.g. clear-cut differentiation between benign and malignant character of the lesion could not be made. There were 40 such cases (1.96%) out of 2332 breast tumors biopsied between 1987-1989. In the cases that proved to be malignant on histology the authors described cytological features which speak in favour of malignancy. In the group of smears which proved to be benign on histology the authors described the features that imitate atypia and should not be misdiagnosed as malignant. The described criteria may be of some help in cytological diagnosis of breast masses.


Acute myelosclerosis--morphological and clinical analysis of the case. A review of literature. A rare case of acute myelosclerosis has been described in a 20-year-old man. Based upon literature data and our own studies a clinical and morphological analysis of the case was presented. The pathogenesis of the disease was also discussed.


Atypical (mild) forms of dihydropteridine reductase deficiency: neurochemical evaluation and mutation detection. We investigated two patients with an atypical (mild) form of dihydropteridine reductase (DHPR) deficiency. Both responded to the loading test with tetrahydrobiopterin; their plasma phenylalanine levels were lowered from 278 mumol/L to 85 and 48 mumol/L and from 460 mumol/L to 97 and 36 mumol/L after 4 and 8 h, respectively. In one of the patients, a combined loading test with phenylalanine followed by tetrahydrobiopterin was also carried out and showed a profile typical for DHPR deficiency. The phenylalanine hydroxylation rate was calculated to be 43 and 87%, 4 and 8 h after cofactor administration, respectively. Diagnosis was confirmed by the absence of DHPR activity in the patient's erythrocytes. In cultured fibroblasts, residual activity of 4 and 10%, respectively, was found. Excretion of urinary pterins was essentially normal, and the biopterin to neopterin ratio in cerebrospinal fluid was increased. Although in both patients cerebrospinal fluid homovanillic acid was found to be normal, and 5-hydroxyindoleacetic acid was substantially reduced, there was no sign of neurologic alterations until the age of 2 y. However, one of the patients recently developed deceleration of head growth, whereas psychomotor development continued to be normal for age. Using the chemical cleavage method on the amplified cDNA, mismatches of T to G at nucleotide 659 and of G to A at nucleotide 475, respectively, were identified. These results also demonstrate that screening for tetrahydrobiopterin deficiency by urinary pterin analysis alone can miss some newborns with mild DHPR deficiency and that all children with tetrahydrobiopterin defects need full neurochemical evaluation together with analysis of the enzyme activity.


[Rhabdoid tumor of the neck].

[Rhabdoid tumor of the neck]. Three pediatric cases of malignant rhabdoid tumor of the neck are described. Clinical data and imaging findings (US, CT and MRI) are stressed. The mass was well defined, containing punctate calcifications in two cases and encasing the vessels in two other cases. Two patients were treated with a chemotherapy regimen according to MMT 89 SIOP protocol, one had chemotherapy and radiotherapy. Two children died of progressive disease; the remaining child who had complete surgical removal of the tumor is on remission 17 months after diagnosis.


Prenatal detection of 45,X/46,XX/47,XXX mosaicism through amniocentesis: mosaicism confirmed in cord blood, amnion, and chorion. Sex chromosome mosaicism in amniotic fluid cells poses a serious dilemma in prenatal diagnosis. Chromosome analysis of 56 primary clones of amniocytes revealed three distinct cell lines. Nine cells (16.1 per cent) demonstrated a 45,X karyotype, 11 cells (19.6 per cent) a 47,XXX karyotype, and the remaining 36 cells (64.3 per cent) had a modal number of 46 chromosomes (46,XX). Cytogenetic evaluation of 100 cells from cord blood, amnion, and chorion following delivery confirmed this triple mosaicism. However, the distribution of the three karyotypes in the pre- and postnatal samples was not found in the same proportions. The cord blood had the most similar frequency to that of the amniotic fluid sample, while the chorion had a significantly increased frequency of 47,XXX cells (41 per cent) and a decreased frequency of 45,X cells (2 per cent). Physical examination of the infant at birth revealed no discernible phenotypic abnormalities. Parental karyotypes were normal. This case highlights the difficulty in determining whether a prenatally detected abnormality will be associated with postnatal phenotypic deviation.


Vitronectin modulates glycosaminoglycan dependent reactions of protein C inhibitor. Protein C inhibitor (PCI), a glycosaminoglycan (GAG) dependent serine protease inhibitor, inhibits its target proteases by forming SDS-stable 1:1 complexes. GAGs alter target enzyme specificity of PCI in such a way that e.g. urokinase (uPA) is the preferred target enzyme in the presence of GAGs while in their absence preferentially tissue kallikrein (TK) complexes are formed. The effect of the GAG-binding adhesive glycoprotein vitronectin (Vn) on the GAG-stimulated inhibition of uPA by PCI was studied using an amidolytic assay. In the presence of heparin, Vn protected uPA from inhibition by PCI in a dose-dependent manner with respect to both, Vn- and heparin-concentration. Vn also was active when heparin was replaced by low-molecular weight heparin or heparan sulfate, respectively. In the absence of GAGs, Vn had no effect on the inhibition of uPA by PCI. In a similar system, Vn was far less effective in modifying the inhibitory function of heparin on the inhibition of TK by PCI. When equimolar concentrations of radiolabelled uPA and TK were incubated with PCI in the presence of heparin, only complexes of PCI with uPA were detectable. Addition of Vn reduced this complex formation, whereas, in contrast, complexes of PCI and TK appeared. These results indicate that Vn modulates both, the activity and specificity of PCI and suggest different structural heparin-requirements for the PCI/uPA versus PCI/TK interaction.


Embryotoxicity and neurotoxicity in rats associated with prenatal exposure to DURSBAN. DURSBAN (DB; active ingredient chlorpyrifos) is a widely-used organophosphate insecticide. The teratogenic and neurotoxic potential of DB was evaluated in rats in utero by exposing embryos on days 0-7 or days 7-21 of development. These prenatal exposures to DB (0.03, 0.1 or 0.3 mg chlorpyrifos/kg, ip) induced physical abnormalities and embryotoxicity. Rat pups which had been exposed to 0.3 mg chlorpyrifos/kg prenatally demonstrated significant behavioral neurotoxicity on postnatal day 16 in the rotorod test compared to time-matched saline-infused litters. Exposure to DB on postnatal day 3, 10 or 12 also caused neurotoxicity as evaluated by the rotorod test. Our studies suggest prenatal exposure to relatively low concentrations of DB may be associated with embryotoxicity, fetal lethality and behavioral neurotoxicity.


Effects of acute ethanol administration on regional serotonin turnover. Numerous studies show the role of biogenic amines in the effects of ethanol. The present work analyzed the effect of an acute sedative-hypnotic dose of ethanol (4 g/kg) on the levels of tryptophan, serotonin (5-HT) and 5-hydroxyindol-acetic acid (5-HIAA), the serotonin synthesis rate and the 5-HIAA elimination rate in several regions of the brain. The 5-HT synthesis rate was not enhanced by ethanol administration, despite increases in brain tryptophan, 5-HT and 5-HIAA, which were primarily in the medulla-pons. The increased indole levels could be due to inhibition of 5-HIAA elimination from the brain.


[Sudeck's disease].

[Sudeck's disease]. The treatment of Sudeck's atrophy is not easy since there is a constant risk of relapse into the inflammatory stage. The earlier the diagnosis is established, the higher is the likelihood that treatment will be successful. As a rule, dystrophic lapses can be avoided by the use of prophylactic measures, even in patients at risk. Here, the cooperation with the pain therapist or anaesthesiologist is required. I am convinced that, by early infusion treatments or drug therapy for the sedation of sympathetic hyperexcitability, improvements in results can be achieved. It stands to reason that psychological guidance for the patient must be taken into due account.


Incubation chambers for the staining of ultrathin sections. An in situ technique especially suited for immunocytochemistry. The preparation and use of incubation chambers for staining of ultrathin sections is described. As an example the immunogold staining is given which can be carried out in these devices in a more simple manner than normally possible, and with good results as shown by electron microscopy.


Cytokeratin and vimentin heterogeneity in human cornea. The body of information on cytokeratin expression in non-epithelial and epithelioid cells is steadily increasing. In this immunocytochemical study using a panel of monoclonal cytokeratin antibodies, we regularly observed cytokeratins no. 8 and 18 together with vimentin in the corneal endothelium of the human eye, but the antibodies exhibited a heterogeneous reactivity pattern. In fetal eye specimens, cytokeratins were already present at the 10th week of gestation, and disappeared at about the 22th week of fetal development. Corneal and conjunctival epithelial cells in the same specimens usually showed uniform cytokeratin 8 and 18 expression, beside the well documented presence of corneal and squamous epithelium type cytokeratins. In 2 of our 7 cases of adults, cells coexpressing cytokeratin and vimentin were observed in the corneal epithelium. The data provide another example of modulation of cytokeratin and vimentin expression, in which simplistic rules cannot be applied.


Localization of NADPH diaphorase, a histochemical marker for nitric oxide synthase, in the mouse spinal cord. NADPH diaphorase-reactive neurons and fibres are present within the entire spinal cord. Moderately to strongly stained neurons were found in the dorsal horn, in particular in lamina III, as well as around the central canal. A dense accumulation of stained neurons was localized in putative preganglionic sympathetic and parasympathetic nuclei. The somatic motor neurons of the ventral horn were found unlabelled. In view of the recently established identity of NADPH diaphorase with nitric oxide synthase, these results suggest that nitric oxide is involved in sensory and autonomic information processing in the spinal cord.


Characterization of two porcine proinsulin reactive monoclonal antibodies by immunostaining of beta-cells in pancreatic sections of different species. Murine monoclonal antibodies against porcine proinsulin were generated by somatic cell hybridization. As detected by radioimmunoassay, 2 monoclonal antibodies KSPI14D4 and KSPI13G10 showed a strong binding to 125I-labelled porcine proinsulin but not to insulin. The species specificity of these 2 monoclonals was found to be different as shown by indirect immunofluorescence using sections of Bouin-fixed pancreata of different species. The KSPI14D4 recognized the proinsulin of pig, mouse, man, cattle, rat, dog, and cat but not that of guinea pig, whereas the KSPI13G10 bound to porcine proinsulin only. From these results it is concluded that KSPI14D4 effectively recognizes a wide-spread epitope located in one of the insulin-C-peptide junctions of the proinsulin molecule, whereas KSPI13G10 is directed to a species-specific epitope of the porcine connecting peptide.


Brain filament proteins in primary cultures derived from chick embryos early in development. Brain filament expression and neurofilament post-translational modifications (phosphorylations) were studied in primary cultures derived from whole 3-4 day chick embryos. After 2-3 days in culture, neurofilament-positive cells formed neuronal aggregates connected by bundles of neurites in a distinctive pattern similar to that observed in cultures derived from embryonal rat brain and neonatal rat cerebellum. Aggregates and neuritic bundles were stained with several monoclonal antibodies reacting with phosphorylated neurofilament epitopes. With two monoclonal antibodies reacting with phosphorylated forms of the high molecular weight neurofilament subunit, staining was only observed after 8 and 10 days in vitro. There was a major difference between rat and chicken with respect to astrocyte differentiation in culture. In chicken, the flat cells surrounding the neuronal aggregates remained constantly GFAP-negative throughout the whole experimental period (10 days). GFAP-positive cells were first observed within the neuronal aggregates on day 8 in vitro.


Perinatal morphine exposure alters peptidergic development in the striatum. It has been reported that perinatal exposure to opiates affects mRNA synthesis, body growth and brain development in mammals, including humans. We have observed that morphine administration in drinking water during the perinatal period alters peptide development in the striatum of the rat. There is a marked increase in substance P and met-enkephalin content, the latter is maintained even at 30 days postnatally. The transient increase or earlier maturation of substance P content is correlated by a more precocious axon terminal organization as revealed by immunocytochemical staining. The increased metenkephalin content is correlated by a higher abundance of preproenkephalin A mRNA and this correlation is particularly evident at 15 days postnatally. At earlier times both northern blotting and in situ hybridization techniques fail to show any significant difference between control and morphine exposed rats, likely because the peptide content is not very different in the two groups or at least the gap is not as wide as at later times.


Influence of cryopreservation on viability and nutritional microcirculation of islets of Langerhans. Isolated pancreatic islets of Syrian golden hamsters were cryopreserved for 1 and 10 weeks, respectively. Following thawing islet mass was found more than 90%. Analysis of viability of these islets by dithizone staining revealed 94 +/- 4% (1 week cryopreservation) and 96 +/- 3% (10 weeks cryopreservation) positively stained islets. After implantation into dorsal skin-fold chambers of syngeneic animals, intravital fluorescence microscopy (5% FITC-dextran, i.v.) showed a regular microvascular network 10 days after implantation. The total diameter of the microvascular network was found similar in cryopreserved islets (day 10: 420.8 +/- 55.2 microns (1 week preservation) and 406.5 +/- 25.2 (10 weeks preservation)) as compared to non-preserved controls (day 10: 411.7 +/- 53.9 microns). Therefore we conclude that cryopreservation of hamster islet isografts for 1 and 10 weeks, respectively, does not alter viability as well as the potential for revascularization. Cryopreservation seems to be an adequate technique for long time storage prior to free transplantation.


Serum protein concentrations in Plasmodium falciparum malaria. In patients with uncomplicated Plasmodium falciparum infection cytokine-mediated serum protein levels of C-reactive protein (CRP), coeruloplasmin (COE), beta 2-microglobulin (B2M), alpha 1-acid glycoprotein (AAG), alpha 1-antitrypsin (AAT), haptoglobin (HPT), prealbumin (PRE), retinol binding protein (RBP), albumin (ALB) and transferrin (TRF) were measured in an endemic area of the Amazonian rain forest. Semi-immune (SI) and nonimmune (NI) patients were investigated. In both patient groups the serum concentrations of CRP, COE and B2M were elevated on admission. In addition AAG and AAT concentrations were increased in NI patients compared to control subjects. Significantly lower serum concentrations of HPT, PRE, RBP, ALB and TRF were seen in both patient groups during the acute phase of the disease, and were more pronounced in NI patients. After a 28-day follow-up, AAT and B2M were normal in SI patients but HPT, AAT and B2M were still significantly altered in NI patients.


[Observation of the posterior urethra by transrectal linear ultrasonography]. Transrectal linear ultrasonography was performed on 59 male patients with dysuria, and voiding movement of the posterior urethra was observed with a videomonitor. Uroflowmetry was performed simultaneously, and the results were compared with those obtained in a previous study. On the image synchronized with maximum flow rate, the diameter of the bladder neck, and the center of the prostatic urethra were measured as parameters and the correlation with maximum flow rate, average flow rate, and residual urine rate, was examined. The forward movement of the ventral side of the posterior urethra was found to play a more important role in the opening of the posterior urethra, than the opening of the bladder neck, and the width of the prostatic urethra to play a more important role in the efficiency of urination than the width of the bladder neck.


[A case of staphylococcal enterocolitis caused by methicillin resistant Staphylococcus aureus]. Recently, the frequency of methicillin resistant Staphylococcus aureus (MRSA) infection has been increasing. We experienced a case of postoperative enterocolitis due to MRSA. The patient was an 81-year-old male with benign prostatic hypertrophy. Urine leakage from the penrose drain tube appeared 1 day after suprapubic prostatectomy. We had used intravenous infusion of antibiotic agents including cefodizine (CDZM), imipenem (IPM/CS) and cefmetazol (CMZ). He developed severe diarrhea, high fever, oliguria, leg edema and ascites 24 days after the operation. MRSA was detected from his feces. Toxic shock syndrome toxin-I (TSST-I) was produced by this bacteria the coagulase type of which was type II. The patient was treated with oral vancomycin (1 g/day), to which this bacteria showed sensitivity and the patient showed, improvement, including symptoms, leucocytosis and serum CRP level 12 days after administration of vancomycin.


Acute phase proteins in "small for dates" babies. II. Haptoglobin, transferrin, alpha-1-feto protein, alpha-1-acid glycoprotein and caeruloplasmin levels. Haptoglobin, transferrin, alpha-1-feto-protein (AFP), alpha-1-acid glycoprotein (AAGP) and caeruloplasmin levels were estimated in 14 "small for dates" (SFD) and 31 "appropriate for dates" (AFD) babies by the single radial immunodiffusion method. The mean caeruloplasmin levels was observed to be significantly reduced in the SFD babies when compared with the AFD babies (t = 3.4582, P < 0.02). None of the other 4 acute phase proteins showed any significant differences in mean concentration between the SFD babies and the controls. The diminished caeruloplasmin levels observed in SFD babies agrees with previous reports in post-natal undernutrition. Our findings of no significant differences in the other 4 acute phase proteins between SFD and AFD babies are however at variance with previous observations of elevated levels of AFP, haptoglobin and AAGP and reduced levels of transferrin in malnourished infants.


Clinical, hematological, and molecular features in Sicilians with Hb S-beta-thalassemia. The clinical, hematological, and molecular features of 81 patients with Hb S-beta-thalassemia and relatives from 76 unrelated families are reported. We analyzed the beta-thalassemia mutations and the beta S haplotypes in all patients and detected 6 different beta-thalassemia alleles: codon 39 (C-->T) (39 cases), IVS-I-1 (G-->A) (12 cases), IVS-II-1 (G-->A) (4 cases), IVS-I-6 (T-->C) (6 cases), IVS-I-110 (G-->A) (14 cases), and IVS-II-745 (G-->C) (6 cases). Eighty patients had haplotype #19 or the Benin type and one had haplotype #17 or the Cameroon type. The type of beta-thalassemia allele had the greatest influence on the phenotypic expression; this was observed for patients with Hb S-beta-thalassemia and for simple beta-thalassemia heterozygotes. The mild IVS-I-6 (T-->C) mutation produced borderline abnormal erythrocytic indices and Hb A2 levels in heterozygotes. Overall, there was a milder expression in beta(S) beta(+) patients (only 7.7% presented severe disease) than in those with the beta(S)beta(0) condition (22.6% had the severe form of the disease).


Clinical course and molecular characterization of a compound heterozygote for sickle hemoglobin and hemoglobin Kenya. We describe a 25-year-old black woman who presented with a long history of anemia requiring transfusions during childhood and adolescence. Molecular analysis revealed her to be a compound heterozygote for the sickle mutation and the approximately 22.7 kb deletion associated with hemoglobin Kenya. This patient's clinical course was more severe than previously reported for the Hb S/Hb Kenya genotype, a probable consequence of concomitant iron deficiency.


Foregoing life-sustaining treatment for adult, developmentally disabled, public wards: a proposed statute. This Article proposes a procedure for making decisions to forego life-sustaining treatment for adult, developmentally disabled, public wards who are not competent to make health care decisions. Few commentators or cases address the special considerations involved in making life-sustaining treatment decisions for this patient population. The proposal attempts to fill this gap with a patient-centered process that allows decisionmakers, without prior judicial approval, to forego life-sustaining treatment for adult, developmentally disabled, public wards who have been reliably diagnosed with specific medical conditions.


Chimeric rat Na,K-ATPase alpha 1/alpha 3* isoforms. Analysis of the structural basis for differences in Na+ requirements in the alpha 1 and alpha 3* isoforms. Na,K-ATPase molecules containing the alpha 1, alpha 2*, and alpha 3* isoforms expressed in HeLa cells exhibit a two- to threefold difference in their K0.5 for Na+ (alpha 1 = alpha 2* < alpha 3*). To investigate the structural basis for this difference, chimeric alpha 1/alpha 3* isoform cDNAs were constructed and expressed in HeLa cells. Na,K-ATPase containing each alpha isoform chimera was analyzed for its Na+ dependence properties. Results of these experiments do not reveal a region in the alpha 1 or alpha 3* isoform that is clearly responsible for the apparent affinity for Na+. It is possible that molecular interactions involving amino acids that span virtually the entire Na,K-ATPase molecule contribute to the determination of this parameter.


Constraints on models for the folding of the Na,K-ATPase. We have attempted to bring together in graphic fashion the available evidence on the structure of the Na,K-ATPase and the H,K-ATPase. There appears to be much room for modification of the existing models for transmembrane folding. More sites on each side of the membrane need to be identified. Whether these will be antibody epitopes, sites of covalent modification, or tags inserted by mutagenesis is less important than that there be many of them and that each be verified by alternative approaches. If any single principle has emerged from the study of the topography of membrane proteins, it is that it is easy to reach conclusions too soon.


Interactions of monocrotophos and quinalphos with Anabaena torulosa isolated from rice soil. The interaction of insecticide combinations of monocrotophos and quinalphos on Anabaena torulosa, by the criteria of absorbance (OD) and packed cell volume (PCV) of the culture, content of chlorophyll a, phycocyanin, carotenoids, total protein, DNA and RNA, heterocyst differentiation and nitrogen fixation, were assessed. In general, monocrotophos and quinalphos, in combination, interacted significantly yielding three different responses viz., additive, antagonistic or synergistic. The nature of the interaction found with OD and PCV was nearly the same for a particular concentration of the insecticides. However, no consistent interaction was observed with respect to carotenoids and phycocyanin. The three types of interaction were noticed for total protein, DNA and RNA. Interestingly, the insecticide combinations at lower concentrations yielded all interaction responses for heterocyst frequency and nitrogenase activity. But, higher concentrations, in combination, resulted in synergism for heterocyst differentiation and nitrogen fixation.


Specific incorporation of kinetin into eukaryotic and prokaryotic transfer ribonucleic acid molecules. We show that kinetin, a non-natural product with strong cytokinin activity, is incorporated into prokaryotic and eukaryotic tRNAs in the exchange reaction catalysed by a putative tRNA-kinetin transglycosylase. We also show that kinetin is specifically incorporated into E. coli tRNA(Tyr) and most probably at position 37. To our knowledge, this is the first report of a nucleic acid base exchange reaction occurring at this position.


Physiological activities of S-(1,2-dicarboxyethyl)glutathione as an intrinsic tripeptide present in liver, heart and lens. S-(1,2-Dicarboxyethyl)glutathione (DCE-GS, CAS 1115-52-2) found in rat liver, heart and lens in considerable amounts, showed an anti-inflammatory effect, which was evaluated by testing the inhibition of the experimental conjunctival edema of rats. An intravenous injection (3 mg/kg) prior to the carrageenan injection prevented the conjunctive edema formation by up to 30%. This peptide also inhibited the histamine release from rat mast cells induced by the compound 48/80. The peptide was added to the mast cells before addition of the compound 48/80 and an inhibition of the histamine release up to 96% at a 1 mmol/l concentration occurred. Furthermore, it displayed an antianaphylactic effect in rats using antibody against chicken egg albumin. An injection of the peptide (30 mg/kg) prior to the antigen administration inhibited color deposition up to 43%. Analogues or derivatives of DCE-GS were synthesized and tested for those inhibitory activities. However, there was no other peptide having stronger effects than DCE-GS and little structure-activity relationship among them.


Coexpression of vimentin, cytokeratin and S-100 in monomorphic adenoma of salivary gland; value of marker studies in the differential diagnosis of salivary gland tumours. An unusual coexpression of glial fibrillary acid protein (GFAP), keratin and vimentin occurs in pleomorphic adenoma of salivary gland. We designed this study to see if coexpression of the markers was also present in monomorphic adenoma of the salivary gland and whether monomorphic adenoma could be distinguished from other salivary gland tumours by marker studies. Immunocytochemical markers were used on fine needle aspiration samples from four cases of monomorphic adenoma, two cases of oncocytic adenoma, three cases of adenoid cystic carcinoma and four cases of pleomorphic adenoma. While positivity for cytokeratin, vimentin and S-100 was consistently found in all cases of monomorphic adenoma, only cytokeratin was expressed in adenoid cystic carcinoma. In pleomorphic adenoma, GFAP, cytokeratin and vimentin were coexpressed while in cases of oncocytic adenoma none of the markers was localized. Thus, it appears that by using a combination of GFAP, cytokeratin, vimentin and S-100 a distinction between these neoplasms may be possible. However, a larger study is needed to establish the usefulness of this approach.


Inhibitory effect of substituted dextrans on MCF7 human breast cancer cell growth in vitro. Substituted dextrans can reproduce some of the properties of heparin and can thus be used to alter cellular growth. We studied the effect of heparin (H108), dextran (D), carboxymethylbenzylamide dextran (CMDB) and carboxymethylbenzylamide sulfonate dextran (CMDBS) on the growth of human mammary cells of the MCF7 tumor line. The cells were cultured in minimum Eagle's medium containing 2% fetal calf serum without biopolymer, or with increasing concentrations of H108, D, CMDB or CMDBS. Growth curves were accurately based on cell counting using a Coulter counter. Cell distribution in the various phases of the cycle was analyzed by flow cytometry. Dose-dependent growth inhibitory effects (400-4000 micrograms/ml) were observed. The effect on MCF7 tumor cells was most apparent with CMDBS. The percentage of cells in the S phase decreased with preferential blocking in the G0/G1 phase. Pre-clinical studies can be anticipated as there is an absence of in vivo toxicity.


Bleomycin but not its derivatives inhibits the in vivo shedding of a rat tumor-associated antigen. Rat fibrosarcoma (KMT-17) cells and their in vitro clone, 10% FCS A3 cells, shed a tumor-associated antigen (TAA), CE7, from the cell surface under growth-promoting conditions. We treated cells with the antitumor agent bleomycin (BLM) and its analogs peplomycin and liblomycin in vitro and in vivo in an attempt to increase expression of this antigen and induce an antitumor response. Although all three agents enhance antigen expression in vitro, proportionate to their direct antiproliferative effects, only BLM enhances antigen expression in vivo. The in vivo regulation of CE7 expression appears not to be related to the direct cytotoxic effects of the antitumor agents but rather to the immuno-augmenting effects of BLM.


[Complex studies of virology, microbiology, biochemistry and epidemiology in the adults of a semi-closed community vaccinated against influenza with the inactivated NIVGRIP vaccine intranasally]. A study was conducted on the specific and non specific immunomodulator effects of the inactivated vaccine NIVGRIP. The results revealed the seroconversion of HAI antibodies against influenza and mumps viruses and the reduction of viral and microbial flora of the pharynx. The kinetics of some proteins specific to the acute stage (siderophiline antitrypsin, microglobulin, complement C1q and C2 factors, C reactive protein and fibronectin) was also investigated and significant quantitative modifications were noticed in vaccinated subjects. The mean illness duration (acute respiratory disease) was three times as short in vaccinated subjects as in controls.


Immunohistochemical investigation on the presence of chondroitin sulfate in calcification nodules of epiphyseal cartilage. Chondroitin sulfate localization in mouse epiphyseal cartilage was studied using CS-56 monoclonal antibody immunospecific for the glycosaminoglycan portion of the molecule. For light and fluorescence microscopy, decalcified specimens were embedded in paraffin, Lowicryl, or were frozen and cryostat-sectioned, and the antigen-antibody reaction was demonstrated by treating sections with IgM-peroxidase, IgM-alkaline phosphatase, or IgM-fluorescein conjugates. For electron microscopy, decalcified and undecalcified specimens were embedded in Lowicryl; ultrathin sections from undecalcified specimens were decalcified by flotation on EDTA; sections from both types of specimens were treated with IgM-immunogold conjugate for demonstration of CS-56 reaction. Before immunoreaction, part of all decalcified sections were digested with Streptomyces or testicular hyaluronidase. Control sections were treated with either mouse and goat non-immune serum, or mouse monoclonal antiserum to human dendritic reticulum cells. Both light and electron microscopy show CS-56 reaction with cytoplasmic components of maturing and hypertrophic chondrocytes. Under the light microscope, immunoreaction was not visible in calcified matrix, and was visible in uncalcified matrix only after hyaluronidase digestion. Under the electron microscope, it was evident both in uncalcified and calcified matrix, although the latter showed few immunogold particles, usually placed on areas which appeared incompletely calcified. Gold particles were chiefly distributed at the periphery of calcification nodules and fully calcified matrix. These results show that CS-56, besides reacting with cytoplasm of maturing and hypertrophic chondrocytes, binds to crystal ghosts and other components of cartilage matrix, immunoreactivity decreasing as calcification increases. This suggests that chondroitin sulfate molecules are either degraded during calcification, or segregated into macromolecular complexes, or both degraded and segregated. The second possibility is supported by the increase of immunosensitivity induced by hyaluronidase digestion.


Immunohistochemical and electron microscope studies of rat islets of Langerhans one month after adult thymectomy. Ultrastructural and quantitative immunocytochemical studies of rat pancreata were carried out 1 month after adult thymectomy. The proportions of insulin-, glucagon-, and somatostatin-immunoreactive cells in the pancreas were estimated on paraffin sections using the unlabelled peroxidase-antiperoxidase method. Relative islet volume, islet size and number were determined on hematoxylin and eosin stained sections. A moderate increase of the islet volume on account of size was found in the pancreas of the thymectomized rats. The proportion of insulin-immunoreactive cells was also elevated. Ultrastructural studies showed a rich supply of secretory granules in most beta-cells. Mixed beta-endocrine-acinar cells were often observed. Mitotic figures were found in single beta-cells. The blood glucose level was in the normal range. The findings suggest a moderate stimulation of beta-cell secretory activity after thymectomy which is not associated with elevated blood glucose levels.


Nerve growth factor receptor immunoreactivity in non-nervous structures of the adult rat brain. The non-neuronal distribution of nerve growth factor receptors (NGFr) in the adult rat brain was investigated immunohistochemically using an anti-NGFr monoclonal antibody (192-IgG). In addition to the neurons known to be NGF-dependent or containing NGF binding-sites, a moderate to strong NGFr immunoreactivity was observed in several non-nervous tissues such as the ependymal cells, the epithelial cells of the choroid plexus, the leptomeninge and the cerebral blood vessels. Vascular immunoreactivity for NGFr occurs in the adventitia and muscular layers of the large arteries (circle of Willis) and, apparently, in all the wall layers of the intermediate or small (pial-arachnoid) arteries. The present results provide direct evidence for the presence of non-neuronal NGFr in the brain of adult rats, and suggest a possible involvement of NGF in roles other than those classically thought.


GFAP-immunoreactive perivascular glia in the chick optic tectum. Immunocytochemical staining of the glial fibrillary acidic protein (GFAP) was utilized to characterize the processes of the astrocytes enveloping the vessel wall in the central nervous system. The study was carried out in the mesencephalic lobes of 18 and 20 incubation-day chick embryos and of 20 day chickens. A perivascular GFAP positivity was mainly detectable in the vessel portions running within the tectum white layers, while it was scarce, or absent, in the grey ones. The perivascular GFAP negativity in the tectum cellular layers was not considered result of the absence of astrocytic endfeet since our previous electronmicroscopical studies evidenced an almost complete perivascular astrocytic ring throughout the tectum layers at hatching time. Present data rather suggest that the expression of the GFAP-made intermediate filaments in developing astrocytes might be controlled by the surrounding microenvironment.


Brain natriuretic peptide (BNP)-like immunoreactivity in the central nervous system of the crested newt. The distribution of brain natriuretic peptide (BNP)-like immunoreactivity (ir) was studied in the brain of a urodele amphibian, the crested newt Triturus carnifex Laur. BNP-like immunoreactive neurons were found mainly in the caudal hypothalamus (retro- and supra-chiasmatic areas) and in the preoptic area. A widespread innervation throughout the brainstem as far as the spinal cord was also observed. By double immunostaining (after section incubation with a-BNP and a-tyrosine hydroxylase-TH-antibodies), close topographical relationships between BNP-like and TH-like immunoreactive neurons within the hypothalamus were found.


Comparative study of glial fibrillary acidic protein (GFAP)-like immunoreactivity in the retina of some representative vertebrates. A polyclonal glial fibrillary acidic protein (GFAP) antiserum was used to study the distribution of GFAP-like immunoreactivity in the retina of adult vertebrates (teleosts, amphibians, reptiles, birds and mammals). GFAP-positive Müller cells were demonstrated in all the species studied, although with different degrees and patterns of immunoreactivity. In nonmammalian vertebrates, Müller cells were the only immunoreactive retinal elements. The staining was located throughout the retina of the species examined, with the exception of the rabbit, which exhibited regional variability in the expression of GFAP. The data indicate that GFAP expression in retinal Müller cells is a common feature of a wide variety of adult vertebrate species.


On the expression of cytokeratins and their distribution in some rabbit gland tissues. The composition and distribution of cytokeratins were studied in a series of exocrine and endocrine glands in the rabbit, by means of anti-human monoclonal antibodies. Rabbit cytokeratins are known to be homologous to those in man, and their in situ distribution was also found to be similar. In acinic cells of the exocrine glands, only cytokeratins 8 and 18 were detected, whereas in ductal cells 1, 5, 10, 11 and 13, 19 were also present. The myoepithelial cells surrounding acini and basal duct cells were stained with the antibodies directed against cytokeratins 13 and 1, 5, 10, 11. Among the endocrine glands, a higher reactivity was observed in the thyroid gland and pancreatic islets, while pituitary and adrenal gland often remained unstained. Although the expression of cytokeratins was found to be almost homogeneous in each glandular cell type, a certain variability was observed, depending on the organ examined and the treatment carried out.


An immunohistochemical study of the distribution of ABH antigens in human submandibular glands at the light and electron microscopic levels. The distribution of blood group antigens ABH in submandibular glands was studied at light and electron microscopy levels by applying ImmunoGold Silver Staining (IGSS) and post-embedding ImmunoGold (IGS) methods, respectively. In IGSS treated samples, a cytoplasmic and a surface form of antigen localization were discernible in the glandular parenchyma. The former was restricted to most mucous cells and to scattered serous cells: A and B antigens were demonstrated in mucous cells of A and B type glands, while H antigen appeared in most mucous and occasional serous elements regardless of the blood type of donors. The latter appeared as a strong H reactivity on cell surfaces of serous acini and ducts regardless of the patient blood type. The IGS method was applied both on non-osmicated samples embedded in LR White resin and on osmicated, Epon embedded samples. In non-osmicated tissues, antigen labelling was revealed in secretory granules and cell surfaces. Positive secretory granules were found in most mucous cells and occasional serous, intercalated, and striated duct cells. A and B antigens weakly reacted in mucous cells of A and B type glands, respectively, while strong H reactivity was seen in mucous, serous, intercalated and striated duct cells of glands of all types. Surfaces labelled with H antigen were found on both lumenal and basolateral membranes of striated ducts in glands of all types. IGS method applied on osmicated, Epon embedded samples, selectively revealed blood group antigens in secretory granules of serous cells but not in the apical vesicles of striated ductal cells. Cell surfaces were completely unreactive.


Role of mast cells in health: daily rhythmic variations in their number, exocytotic activity, histamine and serotonin content in the rat thyroid gland. A chronobiologic transverse study on rat thyroid has been carried out to investigate whether mast cells and their content in biogenic amines normally undergo daily variations and whether these are related to circadian activity of the gland. The mean number of mast cells per microscopic field presents daily variations ranging from 10.9 +/- 3 to 14.6 +/- 3.8 in males and from 8.4 +/- 1.9 to 14.8 +/- 3 in females: these variations show a circadian trend in both sexes, with a 12 hrs period and two peaks at about 11:10/23:10. The mean percentage of degranulated mast cells per microscopic fields shows daily variations ranging from 51 +/- 11 to 60.4 +/- 14.2 in males and from 49.8 +/- 12.5 to 58.3 +/- 13.6 in females; these variations present a circadian rhythm with a 24 hrs period and a mean peak at 02:00. The histamine content of the gland varies in 24 hrs from 20.93 +/- 1.19 micrograms/g w w to 38.08 +/- 1.7 micrograms/g w w, without any sex-related difference: these variations show a rhythmic trend with a 12 hrs period and two peaks at 09:10/21:10. Serotonin content of thyroid presents circadian variations from 15.98 +/- 0.83 to 23.23 +/- 0.61 micrograms/g w w, with a 12 hrs period and two peaks at 04:20/16:20. Whereas the variations of mast cell exocytosis and of serotonin content seem to be chronobiologically linked to circadian variations of gland activity, evaluated on the basis of free and total tetraiodothyronine serum levels, the variations of mast cell number appear to be related to those of thyroid and blood histamine. The present data support the hypothesis that mast cell activity should not be considered as only linked to inflammation or allergic responses.


Enzyme immunoassay for prostate-specific antigen and its diagnostic application in prostate cancer. The purpose of this study was to develop a reliable screening test for prostate cancer. Monoclonal antibodies specific to the prostate-specific antigen (PSA) were generated with an improved hybridoma technique. The hybrid cells were initially cultured in a semisolid medium containing methylcellulose and later transferred to a liquid medium for further subculture. Thirty-six out of a total 1,250 recovered colonies were shown to exhibit a high affinity to PSA by radioimmunobinding assay. Eight hybrid cell lines which secreted either IgG1 or IgG2a antibodies of a high affinity and specificity were established for evaluation. The association constants between PSA and these monoclonal antibodies were shown to range from 1 x 10(9) to 5 x 10(9) M-1. From the results of a matrix cross-matching procedure, pairs of monoclonal antibodies were identified and the corresponding epitopes assigned, and most of them could also be paired with rabbit anti-PSA in a typical sandwich enzyme immunoassay. The designated EIA procedure was performed over 90 minutes at room temperature in a two-stage incubation protocol with a sensitivity of 0.4 ng/mL. The EIA kit was shown to have little cross-reactivity with thyroid stimulating hormone, alpha-fetoprotein, carcinoembryonic antigen and prostate acid phosphatase. Preliminary evaluations with clinically defined patients' sera revealed that proper selections of antibody pairs in sandwich immunoassays are crucial to the adequate performance of the EIA kits.


Modulation of extracellular matrix glycoproteins production by in vitro interacting conditions between rat colonic fibroblasts and tumoral cells. Mixed cultures of fibroblasts with rat colon carcinoma cell lines were used to investigate the production of extracellular matrix glycoproteins. Tumoral cells were shown to influence their production in different ways depending on the cell clone (PROb cells which in vivo produce progressive tumors and REGb cells which produce regressive ones) but also on the relative proportions of stromal and tumoral cells. When fibroblasts were predominant, the REGb cells containing mixture produced higher levels of all protein studied as compared with the PROb cells containing system. When the situation was reversed in favor of tumoral cells, REGb cells containing cocultures still produced more fibronectin, laminin and undulin, but the difference with PROb ones was reduced. On the opposite, cocultures enriched with PROb cells made more entactin and SPARC and approximately equal amounts of tenascin.


In vitro angiogenesis in fibrin matrices containing fibronectin or hyaluronic acid. Angiogenesis involved numerous interactions between extracellular matrix and endothelial cells which may exhibit changes in actin filament distribution. Using an in vitro model, capillary endothelial cells were grown in fibrin matrix containing fibronectin or hyaluronic acid. Actin filament distribution, nucleus localization and cell morphology were observed. Preliminary study showed the formation of tube-, branche- and capillary-like structures within fibrin. In the presence of both fibrin and fibronectin, cells with actin filament stress fibers were more spreading than those in fibrin. In the presence of hyaluronic acid, tubes were limited in extension into the fibrin. In addition, the study of co-localization of nucleus and actin filaments showed different cell behaviours. Migratory cells seem to arrange in parallel to each other and a capillary-like structure may be formed at the proximal extremity of this cell pattern.


Selective beta 1 and beta 2 adrenoceptor blockade on epinephrine-induced arrhythmias in halothane anaesthetized dogs. Beta 2 as well as beta 1 adrenoceptors have been recognized in the heart of vertebrates. They mediate a positive chronotropic action of catecholamines. We compared the effect of selective beta 1 and beta 2 adrenoceptor antagonists on the genesis of halothane-epinephrine arrhythmias in dogs. The arrhythmogenic dose (AD) of epinephrine was increased in the presence of l-metoprolol, a selective beta 1 antagonist (8.40 +/- 1.13 micrograms.kg-1 x min-1; mean +/- SEM), compared with control value (2.62 +/- 0.56) (P < 0.05). In contrast, ICI-118,551, a selective beta 2 antagonist, did not change the AD (2.36 +/- 0.43). Adding ICI-118,551 to l-metoprolol did not affect the AD of epinephrine in the presence of l-metoprolol alone (6.34 +/- 0.74 vs 8.40 +/- 1.13). These results suggest that selective beta 1 blockade is effective in preventing halothane-epinephrine arrhythmias, but selective beta 2 blockade is not.


[The influence of acupuncture on interleukin 2 interferon-natural killer cell regulatory network of kidney-deficiency mice]. The influence of acupuncture on IL2-IFN-NKC regulatory network was investigated, experiments were performed on kidney-deficiency mice received the treatments of acupuncture of "Zusanli" point. The results showed that the interleukin: (IL:), natural killer cell (NKC) activity and interferon (IFN) from the kidney-deficiency mice were lower than that from the normal mice. Acupuncture increased the levels of all the IL2 and NKC activity, acupuncture promoted Newcastle disease virus (NDV) inducing the IFN of kidney-deficiency mice, but also induced the IFN directly. The results suggests that acupuncture could make positive adjustment to IL2-IFN-NKC regulatory network. It provided new theoretical basis for the principles of acupuncture theory on IL2-IFN-NKC regulatory network put forward in recent years.


Recombinant human G-CSF and retinoic acid in synergistically inducing granulocyte differentiation of human promyelocytic leukemic cells. The effects of recombinant human G-CSF (rhG-CSF) and retinoic acid (RA) were studied on the proliferation and differentiation of HL-60 cells and human acute myeloid leukemic cells. Synergistic effect on granulocyte differentiation was observed when HL-60 cells and primary acute promyelocytic leukemic cells were cocultured with RA plus rhG-CSF. rhG-CSF combined with RA increased more significantly the percentage of mature cells than RA alone and greatly increased NBT reduction activity (P < 0.001). These results suggested that proliferated effect of rhG-CSF on leukemic cells may be important for inducing differentiation of myeloid leukemic cells. But this effect might expose the patients to the risk of acute myeloblastic leukemia if G-CSF was used alone. However, RA could not only rule out the latter situation but retain former merit as well. The authors suggest that the combined use of G-CSF with RA is probably a new approach to the treatment of leukemia.


Pseudomonas pseudomallei and melioidosis in China. Though melioidosis has been reported since 1912, it is still a new problem for most Chinese physicians and medical scientists. Because of the lack of understanding and clinical experiences in melioidosis, none had been reported until 1990 in the mainland of China. In view of China's open policy and health work the natural foci of Pseudomonas pseudomallei in South China, ie. Hainan, Guangdong and Guangxi Provinces, should be given sufficient concern. Efforts should be made in training physicians in the endemic areas to improve their knowledge about the disease and to avoid misdiagnosis.


[Distribution of bone morphogenetic protein in human bone and tooth germs: analysis of specificity of monoclonal antibody against bone morphogenetic protein]. A cell strain stably secreting monoclonal antibody against BMP was obtained by hybridoma technique. The monoclonal antibody specifically reacted with the cells of human bone and tooth germs on paraffin embedded tissue sections. Immunohistochemical staining shows that BMP is distributed along collagen fibres in normal bone, also exist in osteoid tissue of new bone, in osteoblasts and in the cells of bone marrow. BMP may be found in human tooth germs such as in predentin, in the cells of outer and inner enamel epithelium, in the cells of dental sac generating alveolar bone. The results demonstrate that generation and growth of human bone and development of tooth germs have relation to BMP. The results at the molecular level prove that some antigenic determinants of human and bovine BMP are the same. BMP function activity inhibition test suggests that this antibody may block the function group of BMP. The ability of the monoclonal antibody to detect antigen and to inhibit generation of new bone makes it potentially useful in purification of BMP and in treating osteosarcoma and other bone tumors.


Preclinical and clinical pharmacology of vinca alkaloids. Vinca alkaloids, including vinblastine, vincristine, vindesine and vinorelbine, are widely used antineoplastic drugs, either as single agents or in combination with other drugs. The mechanism of action of these cell cycle-dependent agents is the inhibition of tubulin polymerisation into microtubules. Numerous studies have been conducted in animals and humans, using various in vivo and in vitro models, to investigate the pharmacological behaviour of this class of antitumour drug. Studies in cellular pharmacology demonstrate that vinca alkaloids are transported by multiple mechanisms, including passive diffusion and energy- and temperature-dependent active transport systems. Moreover, active efflux of drug is involved in the P-glycoprotein-mediated multidrug resistance to vinca alkaloids. This phenomenon may be modulated, in vivo and in vitro, by calcium antagonists and calmodulin inhibitors. The clinical pharmacokinetics of vinca alkaloids after intravenous bolus injection, continuous infusion and oral administration are characterised by a large apparent total volume of distribution, high total plasma clearance and long terminal elimination half-life. Biliary excretion is the main elimination pathway, with low urinary excretion. Pharmacokinetic parameters of vinca alkaloids are time- and dose-dependent, and large inter- and intra-individual variabilities have been observed. Human hepatic P-450IIIA cytochromes are involved in the metabolism of vindesine, vinblastine and probably other vinca alkaloids. Therefore, the possibility of drug-drug interactions must be considered when coadministering drugs in combination cancer chemotherapy. Development of newer semisynthetic analogues of vinca alkaloids and conjugation of vinca alkaloids with monoclonal antibodies may result in derivatives with increased antitumour activity and less clinical toxicity.


Systemic therapy of advanced breast cancer. Many cytotoxic agents have demonstrated activity in advanced breast cancer, the more active agents being cyclophosphamide and the anthracyclines doxorubicin and epirubicin. Combinations of drugs are generally superior to single agents in terms of response rate, duration of response and survival. The treatment of advanced breast cancer can be continued either until treatment failure, or for a limited time from either initiation of therapy or from the observation of complete response. Although these are issues of significant concern, data from randomised trials are limited, and so the question of optimal treatment duration remains open. Randomised trials comparing regimens that differ by a dose intensity factor of less than 2 have failed to demonstrate significant differences in efficacy between the dose levels. With higher doses, as applied in combination with colony-stimulating factors and bone marrow transplantation, response rates seem to increase, but whether this translates into improved survival has not yet been answered by the results of randomised trials. Approximately 30% of patients respond to endocrine therapy. From the results of randomised trials, which have compared the efficacies and toxicities of different endocrine modalities including combined endocrine therapy, single-agent tamoxifen is generally considered as the preferred first-line treatment, leaving progestins and aromatase inhibitors as alternatives for second-line endocrine therapy in responders. In the majority of trials, chemotherapy combined with endocrine therapy has given improved response rates compared with chemotherapy alone, but the differences have not generally been translated into prolonged survival with combined modalities. This gives rise to the question of the optimal sequence of chemotherapy and endocrine therapy, a subject needing further evaluation in future trials.


A review of the antitumour activity of vinorelbine in breast cancer. The use of vinorelbine 30 mg/m2/week as a single-agent treatment in advanced breast cancer has achieved response rates of > 20% as second-line treatment and 40 to 50% as first-line treatment. The major toxicity of the drug is reversible neutropenia; 35 to 50% of treated patients have grade IV neutropenia. The agent did not induce thrombocytopenia and proved mildly emetogenic and neurotoxic. Activity was confirmed in combination with fluorouracil or doxorubicin, when response rates ranging from 60 to 74% were achieved. Thus, vinorelbine appears to be a promising agent in the treatment of advanced breast cancer.


The current and future place of vinorelbine in cancer therapy. Vinorelbine is a new semisynthetic vinca alkaloid that differs chemically from vinblastine by a substitution of the catharanthine moiety. The antitumour activity of vinorelbine against murine tumours, human malignant cell lines and human tumour xenografts in nude mice is evidence of its powerful cytostatic activity against all tumour types. Phase I and phase II studies of intravenous vinorelbine, administered weekly as a single agent or in combination chemotherapy, have been conducted since 1985. Results suggest that vinorelbine has high activity in non-small cell lung cancer (with an overall response rate of 33 to 65%), breast cancer (overall response rate of 46 to 78%) and cisplatin-resistant ovarian carcinoma (over-all response rate of 16% and 35% with single-agent and combination therapy, respectively). In Hodgkin's disease, vinorelbine as a single agent demonstrates high activity, with overall responses ranging from 34 to 90%. Recent phase II studies assessing vinorelbine administered by continuous infusion or orally show promising response rates; however, further trials are needed to validate these preliminary results.


Current options in the treatment of non-small cell lung cancer. Non-small cell lung cancer (NSCLC) is a leading cause of cancer death in industrialised countries, with an overall 5-year survival rate of less than 10%. Good patient performance status is associated with a significantly improved prognosis, whereas the presence of metastases, particularly to the bone or liver, is associated with a poor outcome. Disease staging has largely been standardised using the tumour, node, metastasis (TNM) system, and also provides significant prognostic information. More recently, several biomarkers have been identified in patients with NSCLC, although their prognostic relevance remains to be established. Surgery, when feasible, is the treatment of choice in patients with NSCLC. However, most patients present with locally advanced disease that is not completely resectable. The use of combination chemotherapy regimens has generally been restricted to patients with metastatic, recurrent or unresectable disease, and several relatively active drugs that are generally used in combination have been identified. Combination chemotherapy either alone, or with surgery and/or radiotherapy, has produced some improvements in response rates and, in the neoadjuvant setting, has allowed complete resection in an increased number of patients with otherwise marginally resectable disease. There is also evidence that this approach may increase both disease-free interval and survival time. Thus, the most active chemotherapy regimens (generally those including cisplatin and a vinca alkaloid) appear to provide some benefit in patients with advanced non-small cell lung cancer, but the identification of more active combinations remains a priority.


Vinorelbine. A review of its antitumour activity in lung cancer. Vinorelbine is a new semisynthetic vinca alkaloid. Data from in vitro and in vivo preclinical studies suggest that vinorelbine is active in both small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). There are at present no clinical data on the activity of vinorelbine in patients with SCLC. The response rate with single-agent vinorelbine in patients with NSCLC was 30% overall in two phase II studies including a total of 153 patients; all responses were partial. Response rates of 14 and 33%, respectively were observed in 2 randomised studies, with 1 response of 41 being complete. Vinorelbine is among the most active single drugs in the treatment of NSCLC, but comparative studies with other vinca alkaloids are lacking. Combinations of vinorelbine with cisplatin have shown response rates of 28 to 33% in 2 studies. The drug is well-tolerated in patients with NSCLC. Leucopenia is a dose-limiting factor, while peripheral neurotoxicity does not appear to be a problem.


A gel of glycosaminoglycans lining the anterior and posterior chambers in man: histochemical evidence at light and electron microscopy levels. A thin mucous layer covers the ciliary body and iris epithelium and becomes thicker in the anterior chamber where it covers the anterior surface of the iris, the chamber angle and corneal endothelial surface. It is especially thick at the chamber angle, where it adopts a meniscus shape with the concavity towards the anterior chamber. Cetylpiridinium chloride along with glutaraldehyde was used to precipitate this layer of mucous substance lining the anterior and posterior chambers of the human eye. The staining of the precipitate by cationic dyes suggests that glycosaminoglycans are main components. The specific characterisation with anti-hyaluronic acid monoclonal antibody labelled with colloidal gold reveals the presence of hyaluronic acid in the precipitate. Long unbranched hyaluronic acid molecules may form the skeleton of a gel able to trap and hold virtually any other macromolecule suspended in the aqueous humour. A possible role of such a gel in the regulation of aqueous humour outflow is discussed.


Occlusion of the central retinal artery and ocular neovascularisation: an indirect association? Ocular neovascularisation is recognised as an uncommon complication of central retinal artery occlusion (CRAO) but has been associated most frequently in those patients suffering from carotid artery occlusive disease. We report the finding of neovascularisation after CRAO in a group of 8 patients, who have presented over the previous 6 years. Four showed no evidence of atherosclerotic carotid disease and only 2 were diabetic. Although the group numbers are necessarily small, it is suggested that these cases support the assertion that ocular neovascularisation may occur following CRAO in the absence of any underlying and predisposing ischaemic or inflammatory condition.


Acidic fibroblast growth factor accelerates the healing of acetic-acid-induced gastric ulcers in rats. Acidic fibroblast growth factor (aFGF) was evaluated for the healing of acetic-acid-induced gastric ulcers in rats. The effect of aFGF on angiogenesis in the gastric ulcer bed was determined by the carmine dye infusion method, while its effect on gastric acid secretion was assessed in chronic gastric fistula rats. Oral treatment with aFGF, in the presence of heparin, reduced (ED50 value = 30.2 micrograms/kg/day) the acetic-acid-induced gastric ulcer area, when assessed 1 week later. aFGF was about 1,333-fold more potent than famotidine for healing such ulcers. At a dose of 200 micrograms/kg/day, aFGF increased the carmine density 3-fold and correspondingly reduced (80%) the gastric ulcer area. Thus, the ulcer healing effect of this agent involves angiogenesis in the gastric ulcer bed. This effect of aFGF appears to be unrelated to an inhibition of gastric acid secretion, as it was ineffective in chronic gastric fistula rats. In summary, oral aFGF significantly accelerates the healing of experimental gastric ulcers in rats. It may be a potent and effective agent for the treatment of peptic ulcers in humans.


A simple radioassay for detection of antithyroglobulin autoantibodies. Radioassay technique for detection of anti-thyroglobulin autoantibodies (ATA) in serum samples using protein-A rich Staphylococcus aureus as an immune complex separating agent was comparable (r = 0.99; n = 42; P < 0.001) to that evaluated with anti-human gammaglobulin (AHGG). The inter- and intra-assay coefficient of variations were lower (7.4 and 2.2% respectively) using Staph. aureus as compared to that observed with AHGG (10.4 and 4.7% respectively). A highly significant correlation was observed (r = 0.53; n = 71; P < 0.001) between per cent bound radiolabel thyroglobulin in radioassay and log reciprocal titre of ATA by haemagglutination method. Radioassay scored more number of positive sera as compared to haemagglutination method for normal controls (6 vs 1; n = 46) and patients of thyroid diseases (151 vs 70; n = 238). Due to the relatively poor stability of tracer on storage larger inter-assay coefficient of variations were observed with the radiolabelled preparations older than 10-12 days. The radioassay method evaluated is sensitive, reproducible and useful in detecting presence of anti-thyroglobulin autoantibodies.


Inhibition of nitric oxide synthesis improves survival in a murine peritonitis model of sepsis that is not cured by antibiotics alone. Inhibition of nitric oxide synthesis was investigated in a murine model of advanced sepsis in which antibiotic therapy alone did not improve survival. Seven hours after receiving a lethal intraperitoneal challenge with live Escherichia coli, mice were given either NG-monomethyl-L-arginine (L-NMMA) intravenously, imipenem-cilastatin subcutaneously or a combination of both. L-NMMA (3-300 mg/kg) or imipenem-cilastatin (10 or 50 mg/kg) given alone did not improve survival; co-administration of L-NMMA and either 10 or 50 mg imipenem-cilastatin/kg improved survival significantly. These findings suggest that nitric oxide contributes to the morbidity associated with advanced sepsis and that nitric oxide synthase inhibition may improve the efficacy of conventional antimicrobial treatment of severe infections.


Hodgkin's disease: summary of twenty years' experience. A retrospective analysis on HODGKIN'S DISEASE (HD) was finalized to see if changing the management and therapy during the years we improved the cure rate of lymphomas and reduced the incidence of side effects due to therapy. Up to twenty years' experience was based in two major therapeutic periods: the first included patients observed between 1970 and June 1980 and the second between July 1980 and December 1987. Significant differences between the two periods were the reduction of splenectomies as staging procedure, the reduction of radiation dose and extension and the sequential use of MOPP/ABVD instead of MOPP alone. The analysis included all patients observed over the twenty years under study by looking to the differences concerning response to therapy, survival, relapse-free survival and major consequences due to therapy, namely death not directly related to lymphoma. 377 pts entered the first period and 193 the second one with a minimum follow-up of 4 years. Significant differences were recorded on CR rate, 80.9% vs 90.5%, respectively (p = 0.0024) and deaths in CR, 15.1% vs 2.6%, respectively (p = 0.000). The overall survival shows a probability of 60% and 83% at 11 years for the first and the second group, respectively (p = 0.000) being the probability of survival of 50% at 20 years for the first group of pts. The probability of being in remission is similarly of 79% and 78%, for the first and second group, respectively. The risk of death in remission accounting all causes not related to lymphoma shows a 17% probability vs 6% at 11 years (p = 0.006) for the first and second group, respectively, being 38% for the former group at 20 years. The most frequent single cause of death in remission was secondary leukemia which was recorded in 14 pts on the group of pts observed between 1970 and 1980, all splenectomized and treated by MOPP and extensive radiotherapy. The modifications of therapy of HD have produced improvements concerning the prognosis of pts; these improvements are due mainly to the reduction of late side effects such as acute leukemia and second solid tumor, and to the increase of remission rate and cure rate of the lymphoma.