Increased levels of trkB mRNA and trkB protein-like immunoreactivity in the injured rat and cat spinal cord. Expression of neurotrophins and neurotrophin receptors was examined with in situ hybridization and immunohistochemical techniques 10 days to 6 weeks after ventral or dorsal funiculus spinal cord lesions in adult rats and cats, lesions that have previously been shown to allow axon regrowth. Strongly elevated levels of trkB mRNA were seen in the scar tissue formed in the white matter after both types of lesions. Only small increases were detected for nerve growth factor, brain-derived neurotrophic factor, neurotrophin 3, neurotrophin 4, trk, and trkC mRNA in response to the injuries. trkB protein-like immunoreactivity was increased in the regions that showed elevated levels of trkB mRNA. EM localized this immunoreactivity to neurons, astrocytes, and leptomeningeal cells. Neurofilament immunolabeling and axonal tracing demonstrated that nerve fibers in the scar tissue were concentrated to areas that showed strong trkB protein-like immunoreactivity. The findings implicate a role for neurotrophin receptors in axonal sprouting and glial reactions in the injured spinal cord.


Carbohydrate cycling in signal transduction: parafusin, a phosphoglycoprotein and possible Ca(2+)-dependent transducer molecule in exocytosis in Paramecium. Parafusin, a cytosolic phosphoglycoprotein of M(r) 63,000, is dephosphorylated and rephosphorylated rapidly in a Ca(2+)-dependent manner upon stimulation of exocytosis in vivo in wild-type (wt) Paramecium. In contrast, the temperature-sensitive exocytosis mutant nd9, grown at the nonpermissive temperature (27 degrees C), does not exocytose or dephosphorylate parafusin upon stimulation in the presence of Ca2+; grown at the permissive temperature (18 degrees C), nd9 cells show a wt phenotype. Parafusin contains two types of phosphorylation sites: one where glucose 1-phosphate is added by an alpha-glucose-1-phosphate phosphotransferase and removed by a phosphodiesterase and one where phosphate from ATP is added directly to a serine residue by a protein kinase and removed by a phosphatase. We show here that, in cell fractions from wt Paramecium, both reactions can be carried out in vitro by using uridine(5'-[beta-[35S]thio])diphospho(1)-glucose (UDP[beta 35S]-Glc) and [gamma-32P]ATP, respectively. The characteristics of these pathways are different. Specifically, in the presence of Ca2+, the amount of UDP[beta 35S]-Glc label in parafusin is reduced. In contrast, identical labeling experiments with [gamma-32P]ATP show that Ca2+ enhances labeling of parafusin. Mg2+ had no appreciable effect on either labeling. Removal of the UDP[beta 35S]-Glc label on parafusin in the presence of Ca2+ correlates with the in vivo dephosphorylation seen upon exocytosis. Incubations with UDP[beta 35S]-Glc were then performed with homogenates and nd9 cell fractions grown at 27 degrees C under the ionic conditions used for wt cells. These labelings were not affected by Ca2+, in contrast to results from wt cells but in accord with those obtained earlier with nd9-27 mutant cells in vivo. Factors responsible for both dephosphorylation and Ca2+ sensitivity were found in the high-speed pellet (P2) in wt cells, suggesting that the putative phosphodiesterase is in this fraction and that the defect in the mutant nd9-27 residues in the Ca2+ activation of the phosphodiesterase. We conclude that the in vivo dephosphorylation of parafusin that occurs upon exocytosis is a dephosphoglucosylation due to removal of the alpha-glucose 1-phosphate and more generally that carbohydrates on cytoplasmic glycoproteins may be cyclically added and/or removed in response to extracellular stimuli.


Quantum aspects of brain activity and the role of consciousness. The relationship of brain activity to conscious intentions is considered on the basis of the functional microstructure of the cerebral cortex. Each incoming nerve impulse causes the emission of transmitter molecules by the process of exocytosis. Since exocytosis is a quantal phenomenon of the presynaptic vesicular grid with a probability much less than 1, we present a quantum mechanical model for it based on a tunneling process of the trigger mechanism. Consciousness manifests itself in mental intentions. The consequent voluntary actions become effective by momentary increases of the probability of vesicular emission in the thousands of synapses on each pyramidal cell by quantal selection.


The role of endogenous atrial natriuretic peptide in resting and stress-induced release of corticotropin, prolactin, growth hormone, and thyroid-stimulating hormone. Our previous studies have shown that stimulation of the anteroventral third ventricle region increases atrial natriuretic peptide (ANP) release, whereas lesions of the anteroventral third ventricle or median eminence block the release of ANP from blood volume expansion, suggesting a critical central nervous system participation in this response. ANP is also produced within neurons that have cell bodies in the rostral hypothalamus and axons that extend to the median eminence and neural lobe. In addition to its natriuretic effect, the peptide can inhibit the release of corticotropin (ACTH) and prolactin, anterior pituitary hormones that are released during stress. To determine the physiologic significance of ANP in the control of basal and stress-induced release of anterior pituitary hormones, highly specific antiserum against the peptide (AB-ANP) was microinjected into the third cerebral ventricle of conscious freely moving male rats to immunoneutralize hypothalamic ANP. In the initial experiment, the antiserum or control normal rabbit serum (NRS) was injected into the third cerebral ventricle to determine the effect of the antiserum on basal release of pituitary hormones. The antiserum had no effect on the concentrations of plasma ACTH, prolactin, or thyroid-stimulating hormone for 3 hr after the injection; however, plasma growth hormone concentration, although unchanged for 2 hr, was markedly elevated at 3 hr. These results indicate that although ANP appears to have no effect on the basal release of the other hormones, it has a physiologically significant inhibitory effect on growth hormone release. The delay of the effect is probably related to the time required for the antiserum to diffuse to the site of action of the peptide, presumably at some distance from the ventricle. Since this effect was demonstrable only after 3 hr, in the stress experiment, the antiserum or NRS was microinjected into the third ventricle 3 hr prior to application of ether stress. The rapid elevation of plasma ACTH in NRS-injected rats was markedly augmented by AB-ANP. Ether also induced a rapid increase in plasma prolactin in the NRS-injected animals, as expected. Contrary to the ACTH response, the maximal increase in plasma prolactin after ether was attenuated in animals preinjected with AB-ANP. In the NRS-injected animals, there was a significant decline in plasma growth hormone after the application of ether that was significantly accentuated by AB-ANP, but this was probably the result of the higher initial levels of plasma growth hormone in the ANP-AB group followed by its disappearance with a half-time similar to that of the NRS-injected group. The decline in plasma thyroid-stimulating hormone after ether stress was unaltered in the animals injected with AB-ANP. The results of these immunoneutralization studies suggest that endogenous ANP does not play a role in thyroid-stimulating hormone release. On the other hand, the endogenous peptide appears to have a physiologically significant inhibitory role in suppressing ACTH release during stress, mediated at least partly by suppression of vasopressin release. Endogenous ANP has a pathophysiologic role in augmenting the prolactin release in stress either by inhibiting release of prolactin-inhibiting factors or, alternatively, by enhancing release of prolactin-releasing factors. Endogenous ANP appears to inhibit resting, without altering stress-induced inhibition of growth hormone release by stimulating somatostatin release and/or inhibiting growth hormone-releasing hormone release or by both actions.


RAD25 (SSL2), the yeast homolog of the human xeroderma pigmentosum group B DNA repair gene, is essential for viability. Xeroderma pigmentosum (XP) patients are extremely sensitive to ultraviolet (UV) light and suffer from a high incidence of skin cancers, due to a defect in nucleotide excision repair. The disease is genetically heterogeneous, and seven complementation groups, A-G, have been identified. Homologs of human excision repair genes ERCC1, XPDC/ERCC2, and XPAC have been identified in the yeast Saccharomyces cerevisiae. Since no homolog of human XPBC/ERCC3 existed among the known yeast genes, we cloned the yeast homolog by using XPBC cDNA as a hybridization probe. The yeast homolog, RAD25 (SSL2), encodes a protein of 843 amino acids (M(r) 95,356). The RAD25 (SSL2)- and XPBC-encoded proteins share 55% identical and 72% conserved amino acid residues, and the two proteins resemble one another in containing the conserved DNA helicase sequence motifs. A nonsense mutation at codon 799 that deletes the 45 C-terminal amino acid residues in RAD25 (SSL2) confers UV sensitivity. This mutation shows epistasis with genes in the excision repair group, whereas a synergistic increase in UV sensitivity occurs when it is combined with mutations in genes in other DNA repair pathways, indicating that RAD25 (SSL2) functions in excision repair but not in other repair pathways. We also show that RAD25 (SSL2) is an essential gene. A mutation of the Lys392 residue to arginine in the conserved Walker type A nucleotide-binding motif is lethal, suggesting an essential role of the putative RAD25 (SSL2) ATPase/DNA helicase activity in viability.


Positive supercoiling of DNA greatly diminishes mRNA synthesis in yeast. In Saccharomyces cerevisiae cells harboring a GAL1 promoter-linked beta-galactosidase gene, the simultaneous expression of Escherichia coli DNA topoisomerase I and inactivation of yeast DNA topoisomerases I and II reduces the cellular level of beta-galactosidase to an undetectable level. Analysis of intracellular mRNA level and the density of RNA polymerase along DNA indicates that this reduction is due to the suppression of transcription and that both plasmid-borne and chromosomally located genes are affected. These results are interpreted in terms of inhibition of transcription in vivo due to positive supercoiling of the DNA template: preferential removal of transcription-generated negative supercoils by E. coli DNA topoisomerase I in the absence of both yeast DNA topoisomerases I and II results in the accumulation of positive supercoils in intracellular DNA. In normal prokaryotic or eukaryotic cells, accumulation of positive supercoils is presumably avoided through the balanced actions of DNA topoisomerases.


Modulation of a transient K+ current in the pleural sensory neurons of Aplysia by serotonin and cAMP: implications for spike broadening. To study the contribution of cAMP to the spike broadening produced by serotonin (5-HT) in the pleural sensory neurons of the tail withdrawal reflex, we utilized two phosphodiesterase-resistant cAMP analogs: the Sp diastereomer of cyclic adenosine 3',5'-monophosphothioate (Sp-cAMP[S]), which activates protein kinase A, and the antagonist Rp diastereomer of cyclic adenosine 3',5'-monophosphothioate (Rp-cAMP[S]), agonist Sp-cAMP[S] was injected into the sensory neurons, it caused spike broadening comparable to that induced by 5-HT. In turn, the cAMP antagonist Rp-cAMP[S] blocked approximately 50% of the 5-HT-induced spike broadening. We next examined the K+ currents that are modulated by 5-HT and determined how these currents are affected by cAMP. Confirming Baxter and Byrne [(1989) J. Neurophysiol. 62, 665-679], we found that 5-HT modulated two currents, an S-type K+ current (IKS) as well as a transient and voltage-dependent K+ current (IKV). Rp-cAMP[S] blocked the reduction by 5-HT of the early phase of IKV in parallel with, and to the same degree (60%), as this inhibitor blocked the IKS and spike broadening. These results support the idea that in the pleural sensory neurons cAMP mediates a significant part of the spike broadening that accompanies short-term facilitation produced by 5-HT and that cAMP can produce spike broadening by modulating both IKV and IKS.


cAMP modulates multiple K+ currents, increasing spike duration and excitability in Aplysia sensory neurons. Enhancement of the defensive withdrawal reflex of Aplysia involves a prolongation of the action potentials of mechanosensory neurons, which contributes to facilitation of transmitter release from these cells. Recent reports have suggested that whereas cAMP-dependent modulation of K+ current increases sensory neuron excitability, a cAMP-independent decrease in K+ current may increase the action potential duration and, thus, facilitate transmitter release. We have tested this proposal using Walsh cAMP-dependent protein kinase inhibitor or activators of the cAMP cascade and found that cAMP plays a major role in the spike-broadening effects of facilitatory transmitter; however, broadening requires higher levels of activation of the cAMP-dependent kinase than does increasing excitability. A steeply voltage-dependent transient K+ current, termed IKV,early, and the slowly activating S-type K+ (S-K+) current are both reduced by activation of the cAMP cascade, although with different sensitivities to the second messenger, enabling excitability and spike duration to be regulated independently. Differences in cAMP sensitivity also suggested that the originally described S-K+ current actually consists of two independent components, a slowly activating component and a time-independent, "steady-state" current that is activated at rest.


Leukotriene C4 uses a probenecid-sensitive export carrier that does not recognize leukotriene B4. The export of leukotriene (LT) C4 from human eosinophils, a carrier-mediated process that is temperature-dependent and saturable, was characterized further in eosinophils and in two human leukemia cell lines that do not present an intact 5-lipoxygenase pathway. In eosinophils, KG-1 cells, and dimethyl sulfoxide (DMSO)-differentiated HL-60 cells, the respective Q10 values for temperature-dependent LTC4 export were 3.7, 3.3, and 3.4 and for energy of activation were 28.2 kcal/mol, 23.0 kcal/mol, and 27.8 kcal/mol (1 kcal = 4.18 kJ). When human eosinophils, KG-1 cells, and DMSO-differentiated HL-60 cells were preloaded with defined amounts of intracellular LTC4 by incubation with LTA4 and with incremental amounts of a glutathione conjugate, S-dinitrophenyl glutathione (GS-DNP) by sequential incubation with 1-chloro-2,4-dinitrobenzene, GS-DNP inhibited the export of LTC4 in a dose-dependent manner. By plotting the ratio of total GS-DNP (cell retained plus released) to the sum of total GS-DNP plus total LTC4 against the percentage inhibition of LTC4 release, IC40 values of 0.839, 0.803, and 0.841 were obtained for eosinophils, KG-1 cells, and DMSO-differentiated HL-60 cells, respectively. When cells preloaded with LTC4 were resuspended in incremental concentrations of the organic acid transport inhibitor, probenecid, there was a dose-dependent decrease in LTC4 release; GS-DNP and probenecid inhibited LTC4 release in a cumulative fashion, whereas neither inhibited the release of LTB4 from preloaded nondifferentiated HL-60 cells. Therefore, LTC4 export from cells of bone marrow origin occurs through a probenecid-sensitive membrane carrier shared by other glutathione conjugates and distinct from the LTB4 carrier export system.


Importance of type I and type II mechanisms in the photodynamic inactivation of viruses in blood with aluminum phthalocyanine derivatives. The relative importance of type I and type II mechanisms in the photodynamic treatment of red blood cell concentrations (RBCC) to inactivate viruses was studied using aluminum phthalocyanine tetrasulfonate (AlPcS4), visible light and quenching or enhancing agents of reactive forms of oxygen. Treatment of a human RBCC with 10-13 microM AlPcS4 and 25-26 mW/cm2 visible light resulted in the rapid and complete inactivation of added vesicular stomatitis virus (VSV). The addition of mannitol, glycerol, reduced glutathione (GSH), or superoxide dismutase (SOD), known quenching agents of type I mechanisms, had little to no effect on the rate of inactivation of VSV. Significant inhibition of VSV kill was observed on addition of tryptophan or sodium azide, known quenchers of type II mechanisms. Additionally, the rate of VSV kill was enhanced in the presence of D2O. Taken together, these results indicate a predominant role of singlet oxygen in the inactivation of VSV on photodynamic treatment of RBCC. The relative importance of type I and type II mechanisms on cellular toxicity was also evaluated. Little, if any hemoglobin release was observed on treatment of human or rabbit RBCC with 10 microM AlPcS4 and 44 J/cm2 of visible light in the presence or absence of the above mentioned quenchers. The effect of the addition of quenchers on the recovery and circulatory survival of treated, autologous rabbit RBCC, labeled with 51Cr, was also assessed.(ABSTRACT TRUNCATED AT 250 WORDS)


Structure-activity studies of photoactivated antiviral and cytotoxic tricyclic thiophenes. The photoactivated antiviral and cytotoxic activities of the naturally occurring thiophene, alpha-terthienyl (1), and 15 synthetic analogues were evaluated against murine cytomegalovirus and Sindbis virus, and murine mastocytoma cells. After irradiation with near UV light, alpha-terthienyl and most of its analogues had significant toxicity, with minimum inhibitory concentrations in the range of 0.02-40 microM. In the absence of near UV irradiation, only one analogue had antiviral activity and five were cytotoxic. The most active analogues were those containing carboxylic acid, hydroxyl, or cyano substituents. Quantitative structure-activity relationship analysis of thiophene phototoxicity suggested that the rate of singlet oxygen production is the primary determinant of antiviral and cytotoxic activities. For phototoxicity against murine cytomegalovirus, a significant role for hydrophobicity was also demonstrated. Tricyclic thiophenes show significant potential for photochemotherapy of viral infections and cancer, and further evaluation in animal models is recommended.


Transformation of a bop-hop-sop-I-sop-II-Halobacterium halobium mutant to bop+: effects of bacteriorhodopsin photoactivation on cellular proton fluxes and swimming behavior. We have transformed Pho81, a Halobacterium halobium mutant strain which does not contain any of the four retinylidene proteins known in this species, with the bop gene cluster to create Pho81BR, a BR+HR-SR-I-SR-II-strain. The absorption spectrum, pigment reconstitution process, light-dark adaptation and photochemical reaction cycle of the expressed protein are indistinguishable from those of native bacteriorhodopsin (BR) in purple membrane of wild type strains. Strain Pho81BR permits for the first time characterization of effects of BR photoactivation alone on cell swimming behavior and energetics in the absence of the spectrally similar phototaxis receptor sensory rhodopsin I (SR-I) and electrogenic chloride pump halorhodopsin (HR). A non-adaptive upward shift in spontaneous swimming reversal frequency occurs following 3 s of continuous illumination of Pho81BR cells with green light (550 +/- 20 nm). This effect is abolished by low concentrations of the proton ionophore carbonylcyanide m-chlorophenylhydrazone. Although BR does not mediate phototaxis responses in energized Pho81BR cells under our culture conditions, proton pumping by BR in Pho81BR cells partially deenergized by inhibitors of respiration and adenosine triphosphate synthesis results in a small attractant response. Based on our measurements, we attribute the observed effects of BR photoactivation on swimming behavior to secondary consequences of electrogenic proton pumping on metabolic or signal transduction pathways, rather than to primary sensory signaling such as that mediated by SR-I. Proton extrusion by BR activates gated proton influx ports resulting in net proton uptake in wild-type cells.(ABSTRACT TRUNCATED AT 250 WORDS)


[Occurrence of antibodies against hepatitis C virus (anti-HCV) in patients on long-term hemodialysis]. The prevalence of anti-HCV antibodies, HBs antigenemia and transaminase level were examined in the 136 patients with chronic renal failure on maintenance hemodialysis, treated in dialytic centers of South Poland. In this group anti-HBs and anti-HBc antibodies were also investigated in 60 and 42 patients respectively. Anti-HCV antibodies were present in 16.7-60.0% (mean = 44.1%) of patients depending on the center. These values considerably exceed the percentage of antigen HBs positive patients but they are lower that the percentage of anti-HBs and anti-HBc positive patients.


Effects of angiotensin II and its selective antagonists on inferior olivary neurones. On the basis of biochemical and autoradiographic studies it has been shown that the inferior olivary nucleus (ION) contains predominantly angiotensin II (Ang II) receptors of the subtype 2 (AT2). In the present investigation we used microiontophoretic techniques to test the effect of Ang II on the spontaneous firing rate of rat neurones in the ION in vivo. Ang II excited the majority of histologically identified ION neurones. Furthermore, the antagonism of this angiotensin-induced excitation by selective angiotensin receptor blockers of subtype 1 and 2 (AT1 and AT2) was examined. The excitation could be blocked by low doses of the AT2-antagonists PD 123177 and CGP 42112A, whereas the AT1-antagonist DuP 753 was ineffective even at high doses. On a few occasions, however, ejection of the AT1-antagonist resulted in a potentiation of angiotensin-induced excitation. The results suggest that Ang II has an excitatory effect on a considerable number of ION neurones and that this effect is mediated by AT2-receptors.


Modulation by neurotensin and neuromedin N of adherence and chemotaxis capacity of murine lymphocytes. The action of neurotensin and related peptides has not been yet studied on lymphocytes, although there are studies indicating the stimulative action of neurotensin, a peptide first isolated from bovine hypothalamus, on different functions of phagocytic immune cells. The present study demonstrates that neurotensin and a related peptide, neuromedin N, increased significantly the adherence and chemotaxis capacity of murine peritoneal lymphocytes, when they were incubated in the presence of neuropeptide concentrations between 10(-9) M and 10(-12) M. With respect to their adherence capacity, neuromedin N showed a slightly higher stimulation than neurotensin at a shorter time. However, both neuropeptides stimulated the chemotaxis capacity in a similar percentage. The study of the action mechanisms of these neuropeptides showed that intracellular cAMP levels were not modified by neurotensin or neuromedin N, but using an extracellular calcium chelator, EGTA (1 mM), and a blocker of calcium channels in endoplasmic reticulum, ryanodine (0.5 mM), we observed that neurotensin and neuromedin N could produce their effects through an augmentation of the intracellular Ca2+ concentration. As adherence and chemotaxis are initial processes of immune response, the results show that both neuropeptides may be physiological modulators of the lymphocyte function.


Maternal pregravid weight, age, and smoking status as risk factors for low birth weight births. The Illinois Department of Public Health, in cooperation with the Centers for Disease Control (CDC), monitors trends in the prevalence of prenatal risk factors that are major predictors of infant mortality and low birth weight (LBW). Analyzed data from CDC are available to the department annually. During 1988, a total of 26,767 records of Illinois women giving birth were submitted to CDC. These surveillance data support the fact that women older than 30 years who smoke and enter pregnancy underweight are at greatest risk of delivering LBW babies. Overall, 13.9 percent of underweight smokers had LBW infants compared with 8 percent of underweight nonsmokers. Prevalence of LBW among underweight and smoking women older than 34 years was much higher (29.6 percent) than among those between ages 30 and 34 (15.2 percent). The prevalence of LBW decreased as the pregravid weight increased among normal weight smokers (10 percent) and overweight smokers (8.6 percent).


Noradrenergic activity in anticipatory nausea. Two studies were conducted to examine the hypothesis that noradrenergic activity is a cause of the anticipatory nausea associated with cancer chemotherapy. In the first study concentrations of plasma 3-methoxy-4-hydroxyphenyl-glycol (MHPG) on day 1 of cycle 5 of initial chemotherapy were significantly higher in patients with than without anticipatory nausea. To determine whether elevated MHPG reflected a clinically significant causative role for noradrenergic activity in anticipatory nausea, we conducted a randomized, double-blind, placebo-controlled, crossover trial of clonidine for anticipatory nausea. At a dose of clonidine that produced significant side effects and reductions of plasma MHPG, anticipatory nausea was improved only marginally. These studies do not support a causative role for noradrenergic activity in anticipatory nausea that can be reduced by clonidine with an acceptable therapeutic index.


[Small-cell lung tumors: is a more accurate staging possible and useful?]. Small cell lung cancer is generally staged as a localized or diffuse disease due to its great invasiveness and quick spread. The authors investigated the advantages of a more accurate staging by TNM system applied to small cell lung cancer. Sixteen patients (12 males and 4 females, mean age 54 years, max 66, min 48) were submitted to a treatment protocol consisting of 6 cycles of chemotherapy over an 18-month period. All patients underwent CT before and after the third and sixth cycles. Disease evolution was evaluated by means of the TNM system; relative to the N parameter, the American Thoracic Society criteria were followed. After completion of the third chemotherapy cycle, CT demonstrated reduction in T in 7/16 cases, while in the extant patients T was unchanged. N decreased too in 7 patients and remained unchanged in the others. CT examinations at the end of the whole treatment protocol demonstrated no changes in T. As for N, CT showed evolution from N0 to N2 in one case and from N3 to N0 in another one, while no changes were observed in the extant patients. The M parameter was constantly negative in all cases. Our results demonstrate that this approach to small cell lung cancer permits a more accurate characterization of the disease, thus making it easier to monitor the positive/negative response to treatment and allowing the latter to be personalized.


Effects of angiotensin peptides on cholinergic neurotransmission in rabbit tracheal smooth muscle. To determine the effects of angiotensins I, II, and III (A I, A II, and A III) on airway smooth muscle functions, we studied isolated tracheal segments from rabbits under isometric conditions in vitro. Addition of A II and A III but not A I potentiated the contractile response to electrical field stimulation (EFS) at 5 Hz in a dose-dependent fashion. The frequency-response curves for EFS were displaced to the left by A II and A III, whereas the contractile responses to exogenous acetylcholine remained unchanged. Angiotensin-induced potentiation of the response to EFS was further increased in the presence of physostigmine. These results suggest that A II and A III may prejunctionally potentiate the vagally-mediated contraction of airway smooth muscle.


Phorbol ester alters carbachol-stimulated diacylglycerol formation in parotid acinar cells through the hydrolysis of phosphoinositides and phosphatidylcholine. Incubation of rat parotid acinar cells with phorbol 12,13-dibutyrate (PDBu) resulted in inhibition of carbachol-stimulated formation of sn-1,2-diacylglycerol (DAG). PDBu pretreatment inhibited carbachol-induced turnover of phosphoinositides; this inhibition was indicated by phosphatidylinositol 4,5-bisphosphate breakdown. This pretreatment also attenuated the effect of carbachol on inositol phosphate generation and phosphatidylcholine hydrolysis. These results show that PDBu alters carbachol-stimulated DAG formation through the hydrolysis of phosphoinositides and phosphatidylcholine.


The FPbase microcomputer system for managing community health screening and intervention data bases. Health promotion and intervention projects at State and community levels need computerized data bases to assist in making policy decisions and in operating the projects. Computer data base systems are used in entering, storing, retrieving, and analyzing information about health project activities and their participants in a timely and cost-effective manner. Computer support is essential for such labor-intensive tasks as post-screening followup of participants, identifying subpopulations, and evaluating recruitment efforts and behavior change programs. The Pawtucket Heart Health Program developed a microcomputer software package, FPbase, for community health project data base management. FPbase is described and is available for use by other organizations. FPbase incorporates formative and process interactive data base activities and is suitable for use in operating intervention and screening programs at State and local levels. The system accommodates management of data for social marketing, evaluation, followup, and promotional activities.


Effects of isoxazolidine or triazolidine on rat serum lipids in vivo and LDL and HDL binding and degradation in human and rodent cultured cells in vitro. 2-(3,4,5-Trimethoxybenzoyl)-4,4-diethyl-3,5-isoxazolidione (TDI) and 1-acetyl-4-phenyl-1,2,4-triazolidine-3,5-dione (APTD) are two chemically related derivatives which have demonstrated potent hypolipidemic activity in mice at 20 mg/kg/day I.P. for 16 days. The purpose of this study is to correlate in vivo effects of TDI and APTD on rat serum lipoprotein lipids and apoprotein levels as well as plasma clearance and tissue uptake with effects of the agents on tissue cultured cells' LDL and HDL receptor binding, internalization and degradation. This study also correlates in vivo effects of TDI and APTD with endogenous enzyme activities regulated by these high affinity receptors. In rats at 20 mg/kg/day orally serum cholesterol, triglyceride, and VLDL-cholesterol levels were effectively reduced while HDL cholesterol levels were significantly elevated with both agents. These compounds in human hepatocytes lowered LDL receptor binding and degradation, whereas HDL receptor binding and degradation were elevated in human hepatocytes, rat small intestinal epithelium cells, human BG fibroblasts, rat aorta cells and mouse macrophages. These drugs inhibited HMG CoA reductase and sn-glycerol-3-phosphate acyl transferase activities, findings consistent with the observed in vivo reductions in serum cholesterol and triglyceride levels. Both drugs reduced activity of acyl CoA:cholesterol acyl transferase and accelerated activity of neutral cholesterol hydrolase in liver and aorta cells. This modulation by the drugs should reduce disposition of cholesterol esters in these tissues especially aorta wall; this effect was indeed observed in vivo. In the presence of TDI and APTD, HDL uptake of intracellular cholesterol from fibroblasts was accelerated. This was consistent with results from in vivo rat studies showing that HDL clearance was faster after treatment while clearance of LDL slowed. Tissue uptake of HDL and LDL after drug treatment was reduced for the major organs; however the liver accumulation was elevated. The accelerated uptake in the liver was probably due to the observed higher levels of Apo-E and Apo-AI in HDL after drug treatment. Increased excretion of cholesterol from the liver to the bile after drug treatment indicated that the reserve cholesterol transport system by HDL was accelerated by the agents in vivo.


National survey of state epidemiologists to determine the status of Lyme disease surveillance. In 1990, the Council of State and Territorial Epidemiologists and the Centers for Disease Control made Lyme disease (LD) nationally notifiable and developed a national case definition. State epidemiologists were surveyed about their State LD surveillance system. Responses were received from all States and the District of Columbia. As of November 1991, LD was notifiable in 45 States. A total of 44 of these 45 States use the 1990 national case definition for case confirmation. Twenty-five State health departments offer diagnostic testing for LD, and 38 States have conducted surveys for infected ticks. This study has shown that there has been greater standardization of LD reporting with the adoption of a new national case definition for LD. However, many States confirm cases using data that are not a part of the criteria used for the national case definition.


Involvement of central GABAA receptor complex in controlling the basal gastric secretory activity in pylorus-ligated rats. Effect of compounds related to gamma-aminobutyric acid (GABA) receptor complex on gastric acid secretion was studied to clarify a role of endogenous GABA in controlling the basal secretory activity in pylorus-ligated rats. When pentobarbital, a GABAA stimulant, was given to pylorus-ligated rats after recovery from ether anesthesia, the drug did not modify gastric acid secretion. In contrast, pentobarbital significantly stimulated acid secretion in rats anesthetized with urethane. The basal acid secretion was much higher in the rats without urethane anesthesia. Diazepam, an agonist of the benzodiazepine (BZP) receptor, did not change gastric acid secretion in both the animal groups. Picrotoxin, pentylenetetrazol (PTZ) and bicuculline, which block GABAergic mediation, significantly reduced gastric acid secretion in rats without urethane anesthesia. Moreover, the inhibition by picrotoxin was reversed completely by pentobarbital and moderately by diazepam, while the inhibition by PTZ was partly reversed by pentobarbital but not affected by diazepam. These findings suggest that the central GABAA receptor complex may play a significant role in controlling the basal gastric secretory activity. The direct involvement of the BZP receptor may be small, if any. The stimulatory mechanism through the mediation of GABAA receptor complex appears to be demonstrable when the basal secretory activity is lowered.


Development and supermarket field testing of videotaped nutrition messages for cancer risk reduction. This article describes the development and field testing of five 1-minute videotaped nutrition public service announcements focusing on the role of nutrition in reducing cancer risks. Topics include dietary fat, vitamin A, vitamin C, cruciferous vegetables, and fiber. Produce departments in two large supermarkets were the test sites. Videotapes were shown over a 3-day period for a total of 20 hours in each store. Of the total 1,196 customers intercepted, 1,050 (88 percent) agreed to be interviewed to determine whether they had viewed the video; the impact of the video on nutrition awareness, diet, and immediate purchases; and their perception of the stores' providing nutrition information. Only 26 percent of the customers viewed the videotapes. The main reason cited for nonviewing was lack of time. Forty-three percent of viewers stated that this was new information to them, and 21 percent said they would change their eating habits as a result of seeing the tape. Seventeen percent of viewers stated that they actually purchased foods recommended in the tapes. Of all the customers surveyed, 40 percent said they would shop more often at a supermarket that showed nutrition videos.


[Benign tumors of the liver. Clinical and radiological data]. Cysts and haemangiomas, which are frequent benign tumours of the liver, must be separated from hepatic adenoma and focal nodular hyperplasia. The former are usually diagnosed by ultrasonography and/or computed tomography (CT), and they exceptionally require surgery. The latter, much rarer, are similar in that both occur in young women and have the same imaging characteristics: CT does not always show the sign that confirms the diagnosis of focal nodular hyperplasia (i.e. a central "scar" vascularized at CT-angiography); scintigraphy would provide the diagnosis, but it is at fault in 30 to 40% of the cases. Excluding a malignant tumour (hepatocellular carcinoma or fibrolamellar hepatocarcinoma) is sometimes difficult. Often more than a certainty, it is a collection of convergent clinical and radiological data that leads to the correct diagnosis.


[Treatment of benign tumors of the liver]. The treatment of benign liver tumors is dependent on the type of lesion, its symptoms and its natural history. Hemangiomas require no therapy in most cases. Surgical resection is indicated in symptomatic and/or complicated cases. Simple biliary cysts usually require no therapy. Rare cases of simple cysts with symptoms and/or complications are treated by percutaneous alcoholization or surgical fenestration. Polycystic liver disease requires specific therapy when the volume of the cysts becomes life threatening. Treatment may include percutaneous alcoholization, surgical fenestration and even liver transplantation in selected cases. Hepatic cystadenomas must be resected because of the risk of malignant transformation. Benign hepatocellular tumors include hepatocellular adenoma and focal nodular hyperplasia. Hepatocellular adenoma must be treated by withdrawal of oral contraceptives and tumour resection because of the risks of hemorrhage and malignant transformation. Focal nodular hyperplasia can be left in place when diagnosis is certain, because it always follows an uncomplicated course. Diagnosis may be confirmed by a surgical biopsy taken at exploratory laparotomy. In cases with doubts or with symptoms, surgical resection is indicated.


[Liver cancer and hepatitis B and C virus]. Chronic infection by hepatitis B and C viruses is frequently associated to the development of primary liver cancer. Liver cirrhosis, induced by these viral infection, plays an important role in the liver carcinogenesis. In addition, HBV has a direct role in liver cell transformation by a transactivating effect of some viral proteins as well as insertional mutagenesis. The role of hepatitis C virus is not known. The strong association, even in France, of primary liver cancer to these viral infections underline the importance of their prevention by vaccination.


[Surgical treatment of primary cancers of the liver]. Hepatocellular carcinoma (HCC) is the most frequent malignant tumor of the liver and is associated with cirrhosis in 90% of cases. It is often a multicentric tumor with vascular involvement but surgical resection, when it is possible, remains the best treatment of HCC. Major resection is feasible in patients without liver insufficiency; patients with mild liver insufficiency can be treated by local resection. In selected cases, liver transplantation is indicated in order to prevent the high risk of recurrence of HCC which is observed after resection.


[Non surgical treatment of hepatocellular carcinoma]. During the last past years, the curative and preventive treatment of hepatocellular carcinoma (HCC) has improved. The regular follow-up of patients with chronic liver disease using ultrasound examination and serum alpha-fetoprotein determination, leads to diagnose hepatocellular carcinoma at an early stage, when curative treatment could be quite efficient. Besides surgery which has been the only hope of cure for a few patients, efficient medical procedures--ie chemoembolization, in situ radiotherapy and alcohol injection--are emerging. The place of these different therapeutic procedures is to be defined. In addition, the prevalence of hepatocellular carcinoma should decrease due to the improvement of preventive policy--ie vaccination against hepatitis B infection and familial screening for genetic hemochromatosis.


Influence of dry heat treatment and anticoagulant on coupling of fibrinogen and fibronectin to von Willebrand factor in normal plasma. In plasma from healthy subjects a coupling was identified between von Willebrand factor (vWf), fibrinogen (fg), and fibronectin (fn) that was dependent of anticoagulants heparin, EDTA, and citrate. Binding was quantitated by ELISA methodologies, based on polyclonal antibodies directed against the proteins studied, in order to express the percentage of moles of fg or fn bound to moles vWf, C[fg/vWf] or C[fn/vWf] (mol/mol)%. The intra-assay coefficients of variation (CV%) for fg and fn bound to vWf were 10.6% and 7.4% (n = 10) respectively, and the inter-assay coefficients of variation were 24.4% and 22.2% (n = 10). The largest degree of coupling was found in heat-treated lyophilized heparin plasma, where C[fg/vWf] and C[fn/vWf] were 12.9 +/- 1.4 (mol/mol)% and 2.4 +/- 0.1 (mol/mol)% (mean +/- SD). Binding was further qualitatively demonstrated through experiments using gel filtration chromatography and agarose gel electrophoresis followed by immunoblotting. In all instances coupling of vWf with fg was higher than with fn. Lyophilisates of normal plasma that were subjected to dry heating (60 degrees C in 72 h) showed considerably increased coupling. Previous investigators, studying reconstituted factor VIII concentrates by means of gel filtration, pointed out that an association between vWf, fg and fn was present in such therapeutic material. This investigation signifies that a coupling between these proteins may be present even in the source plasma, and that 'dry heating' increases binding. Implications of these results are discussed.


Lipolytic effect of low-molecular-weight-heparin (Fragmin) and heparin/dihydroergotamine in thromboprophylactic doses during total hip replacement. Eighteen patients admitted for implantation of a primary hip prosthesis were randomized to one of the following thromboprophylactic regimens: (1) dextran 500 ml 12 h-1 on the day of surgery and 500 ml on days 1, 3, and 5 postoperatively, (2) heparin 5000 IU and dihydroergotamine 0.5 mg 8 h-1, or (3) Fragmin (Kabi-Pharmacia, Sweden) 2500 IU 12 h-1 on the day of surgery and 5000 IU 24 h-1 throughout the observation period. Fragmin is a low molecular-weight heparin which has proven both efficient and safe as a thromboprophylactic drug. The purpose of the study was to elucidate whether Fragmin had a lower lipolytic (fatty acid-mobilizing) effect than standard heparin. The plasma level of lipoprotein lipase (LPL) was increased on the first post-operative day both in the heparin/dihydroergotamine (preoperative 217 +/- 79 microU ml-1 vs. 1289 +/- 318, n = 6, p < 0.05) and the Fragmin-groups (136 +/- 16 vs. 668 +/- 123, n = 6, p < 0.05). This increase in plasma LPL was significantly higher in the heparin/dihydroergotamine-group compared to the Fragmin-group (p < 0.05). In both groups, the enzyme activity gradually decreased during the next 2 days. The circulating level of non-esterified fatty acids increased post-operatively only in the heparin/dihydroergotamine-group. Thus, the lower lipolytic effect of Fragmin compared to standard heparin is detectable using thromboprophylactic doses during surgery in patients on a standard hospital diet. From a theoretical viewpoint, the use of Fragmin is advantageous in patients with myocardial ischaemia and patients risking respiratory distress problems due to a low circulating level of non-esterified fatty acids.


FK 383 DS, a new silica gel for the determination of unsaturated haptocorrin and transcobalamin II in serum. The ability of selective absorption of the polypeptide transcobalamin II by silica gel was used for the determination of the unsaturated cobalamin binding capacity of haptocorrin and transcobalamin II. Two different silica gels, QUSO G 761 and FK 383 DS, were compared by Sephacryl gel-filtration and by determination of unsaturated haptocorrin and transcobalamin II from forty different patient sera. It was found that the silica gels act identically for this purpose.


Haemorrhagic effect of enoxaparin, a low molecular weight heparin. Comparison with unfractionated heparin in humans. The haemorrhagic effects of unfractionated heparin (UFH) and the low molecular weight heparin (LMWH) enoxaparin were investigated and compared in the gastric mucosa (haemorrhage induced by biopsy) and skin (haemorrhage induced by Simplate) of 12 healthy volunteers. Administration of UFH and LMWH (given in a dose of 75 anti-Xa U/kg intravenously) increased median gastric bleeding time (3.5 min) and geometric mean blood loss (11.5 microliters) to 19 min (p = 0.00003) and 54.1 microliters (p = 0.0021) after UFH and to 13 min (p = 0.008) and 29.0 microliters (p = 0.275) after LMWH. Median skin bleeding time (4.25 min) increased to 6.0 min after UFH (p = 0.003) and to 6.75 min after LMWH (p = 0.0008). Mean heparin activity in plasma was 20% higher after LMWH than after UFH. The calculated gastric bleeding time to heparin activity ratio was significantly lower for LMWH than for UFH (p < 0.05).


Localization of targets for anti-ulcer drugs in cells of the immune system. The gastric mucosa consists of the epithelium, which lines the lumen, the lamina propria, and the muscularis mucosae. The targets of drugs used to treat stomach and duodenal ulcers are thought to be the acid-secreting parietal cells of the epithelium. However, immune cells in the lamina propria are the only cells that showed detectable messenger RNAs for histamine, muscarinic, gastrin, and dopamine receptors by in situ hybridization histochemistry. None of the epithelial cells expressed any of these messenger RNAs. Thus, the targets of antiulcer drugs seem to be cells of the immune system in the gut and not parietal cells, as generally believed. This conclusion may revise the thinking about ulcer formation and may shed light on the etiology of such chronic small intestinal diseases as Crohn's disease.


[Changes of subtypes of alpha 1-adrenoceptor in blood vessels of spontaneously hypertensive rats]. The two subtypes of alpha 1-adrenoceptors in blood vessels were compared between stroke prone spontaneously hypertensive rats (SHRSP) and WKY rats in vitro and in vivo. Pretreatment with 50 mumol/L chlorethylclonidine (CEC) for 30 min decreased the maximal contractions induced by norepinephrine (NE) to 31.4 +/- 8.3% and 35.2 +/- 2.9% (aortae); 68.4 +/- 8.2% and 80.1 +/- 7.2% (renal arteries); 68.4 +/- 6.3% and 5.4 +/- 7.0% (mesenteric arteries) of the controls in the SHRSP and the WKY rats, respectively. All the differences between the SHRSP and WKY rats were not statistically significant. In contrast, the blocking effects of nifedipine were much stronger in the SHRSP rats than those in the WKY rats. In the presence of 10 mumol/L nifedipine the maximal contractions induced by NE were decreased to 3.1 +/- 1.5% and 56.5 +/- 4.8% (P < 0.01, aortae); 9.0 +/- 4.1% and 23.6 +/- 3.5% (P < 0.05, renal arteries); 5.9 +/- 2.5% and 28.0 +/- 0.8% (P < 0.01, mesenteric arteries) of the controls in the SHRSP and the WKY rats, respectively. The experiment in vivo also showed that the effects of nifedipine on decreasing basal blood pressure and on antagonizing phenylephrine were increased in the SHRSP rats, as compared to the WKY rats. Analyses of two components of the contractions induced by 10 mumol/L NE in aortae demonstrated that both phasic and tonic contractions were mainly caused by activations of the alpha 1B subtype. There were no significant differences of blocking effects of nifedipine on the phasic contractions between the SHRSP and WKY rats.(ABSTRACT TRUNCATED AT 250 WORDS)


[The objectives, failures and new ways in hospital aftercare and rehabilitation]. One of the main aims of oncological aftercare is to improve the transition from acute into chronic medicine, i.e. from curative into rehabilitative treatment. The somatic, psychic, social, professional, and family problems due to the tumour and its therapy have to be eliminated, improved or prevented. Up to now, oncological rehabilitation often was considered an isolated measure. The lack of cooperation between curative and rehabilitative medicine often resulted in the fact that patients in need of aftercare were not sent to appropriate rehabilitation clinics. There was no feed-back between pre- and after-treating doctors. The recommendations of the commission for further development of rehabilitation in the social insurance (VDR), elaborated together with the study-group for rehabilitation and aftercare and the study-group for the protection of the quality standard in oncology, aim at improving the above mentioned target. Active and forward-looking measures should improve the significance of clinical aftercare. The documentation of rehabilitation needs will allow more differentiated rehabilitation measures and long-term evaluation of their success. For a more effective integration of rehabilitative aspects into primary treatment, there is the demand for a closer cooperation between primary care and aftercare.


Surgical strategy in the management of non-small cell ectopic adrenocorticotropic hormone syndrome. Non-small cell ectopic adrenocorticotropic hormone (ACTH) syndrome is a rare cause of hypercortisolism that may require surgery for either curative resection or palliative adrenalectomy. We report our surgical experience with 41 patients with ectopic ACTH syndrome and no evidence of small cell lung cancer at initial evaluation. All 41 patients had documented hypercortisolism secondary to ectopic production of ACTH. Based on imaging study results, we determined that 21 patients had localized/resectable disease; eight patients had metastatic disease, and 12 patients had occult disease at examination. Of the 21 patients with localized disease, 16 (76%) were cured of ectopic ACTH by surgery (15 bronchial carcinoid, one pheochromocytoma). Patients with bronchial carcinoid had the greatest probability for cure of ectopic ACTH syndrome, and patients with thoracic primary tumor were more likely to be cured than patients with abdominal primaries. Of the eight patients who had metastatic disease, none were cured of the disease; five patients underwent bilateral adrenalectomy, and three patients were given medical therapy. Only one patient was alive after 5 years. Of the 12 patients who had occult disease, four patients were eventually cured of the disease (three bronchial carcinoid, one thymic carcinoid); one patient died of disease (small cell lung cancer), and seven patients still have occult disease. Nine of 12 patients with occult disease underwent bilateral adrenalectomy for surgical management of hypercortisolism. This study suggests that the most common primary focus of ectopic ACTH production is within the thorax with 25 of 34 (74%) identifiable tumors originating within either the thymus or bronchus. Adrenalectomy offers excellent palliation of hypercortisolism secondary to either occult or metastatic disease. Patients who initially have localized disease usually have bronchial carcinoids and have a high probability of cure with surgical resection (81%).


[The clinical significance of detecting the inhibition of topoisomerase I by the sera of patients with systemic scleroderma]. Topoisomerase I activity was studied by electrophoresis in agarose gel according to plasmid DNA relaxation. Sera from 62 patients with systemic scleroderma, 35 with Raynaud's syndrome, 8 with focal scleroderma, 15 with systemic lupus erythematosus, 20 with rheumatoid arthritis and 20 healthy subjects were examined. Out of 62 sera from SSD patients, anti-topoisomerase activity was found in 67.8% of cases. The test appeared positive in 79% of patients with diffuse and 63% with limited disease patterns. The mean age and disease standing were similar in the positive and negative groups. An increase of the skin count and more frequent occurrence of trophic disorders in patients with inhibition of the enzyme were recorded. 40% of the patients demonstrated the coincidence of the results with the use of the topoisomerase test and ELISA. In patients with other rheumatic diseases and in the healthy subjects, no inhibition of the enzyme was found.


[An analysis of the hormonal response during the performance of stress tests in patients with systemic lupus erythematosus and systemic scleroderma]. Patients with systemic lupus erythematosus (SLE), systemic scleroderma (SSD) and donors were examined for the blood levels of adrenocorticotropic hormone, hydrocortisone, follicle-stimulating hormone, luteinizing hormone, prolactin, estradiol, testosterone, progesterone, thyroid-stimulating hormone, triiodothyronine, thyroxin, and insulin. The corticotropin load test was carried out in 38 SLE patients, 32 SSD patients and 24 donors. The prednisolone test was made in 15 SSD patients and 27 donors. The studies were made with the aid of RIA. The patients with SLE manifested a decline of the basal level of hydrocortisone as well as a reduction of the reserve potentialities of the pituitary-adrenal system. The patients with SSD demonstrated a negligible decrease of the basal level of hydrocortisone with an evident lowering of the reserves of the same system. The treatment of SLE and SSD patients with glucocorticoids was followed by marked hyperinsulinemia.


Neutrophil elastase potentiates cathepsin G-induced platelet activation. We have previously demonstrated that human neutrophil cathepsin G is a strong platelet agonist that binds to a specific receptor. This work describes the effect of neutrophil elastase on cathepsin G-induced platelet responses. While platelets were not activated by high concentrations of neutrophil elastase by itself, elastase enhanced aggregation, secretion and calcium mobilization induced by low concentrations of cathepsin G. Platelet aggregation and secretion were potentiated in a concentration-dependent manner by neutrophil elastase with maximal responses observable at 200 nM. Enhancement was observed when elastase was preincubated with platelets for time intervals of 10-60 s prior to addition of a low concentration of cathepsin G and required catalytically-active elastase since phenylmethanesulphonyl fluoride-inhibited enzyme failed to potentiate cell activation. Neutrophil elastase potentiation of platelet responses induced by low concentrations of cathepsin G was markedly inhibited by creatine phosphate/creatine phosphokinase and/or indomethacin, indicating that the synergism between elastase and cathepsin G required the participation of ADP and thromboxane A2. On the other hand, platelet responses were not attenuated by the PAF antagonist BN 52021, signifying that PAF-acether did not play a role in elastase potentiation. At higher concentrations porcine pancreatic elastase exhibits similar effects to neutrophil elastase, demonstrating that the effect of elastase was not unique to the neutrophil protease. While neutrophil elastase failed to alter the ability of cathepsin G to hydrolyze a synthetic chromogenic substrate, preincubation of platelets with elastase increased the apparent affinity of cathepsin G binding to platelets.(ABSTRACT TRUNCATED AT 250 WORDS)


Hepatocellular carcinoma in children with hepatitis B surface antigen. This study discusses four children of hepatocellular carcinoma (HCC) who were asymptomatic HBsAg carriers or had HBsAg-positive chronic hepatitis for 3 to 11 years before the occurrence of the carcinoma. Three of these four patients were positive for anti-HBe at 3 to 5 years before the diagnosis of hepatocellular carcinoma. Autopsy findings disclosed liver cirrhosis in all the four patients. To the best of our knowledge few reports have documented children in HBsAg carrier status or with HBsAg-positive hepatitis prior to the development of hepatocellular carcinoma. It is emphasized that HBsAg-positive children, with or without detectable hepatic lesions in routine examinations, have a possibility of developing HCC, and should be carefully monitored for long periods.


Effects of acyclovir, oxetanocin-G, and carbocyclic oxetanocin-G in combinations on the replications of herpes simplex virus type 1 and type 2 in Vero cells. 9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV) and novel nucleosides, 9-(2-deoxy-2-hydroxymethyl-beta-D-erythro-oxetanocyl)guanine (oxetanocin-G, OXT-G) and (+)-9-[(1R, 2R, 3S)-2, 3-bis(hydroxymethyl)cyclobutyl]guanine (carbocyclic oxetanocin-G, carbocyclic OXT-G) possessed substantial antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). ACV inhibited only viral thymidine kinase positive (TK+) herpes viruses, although the latter two compounds inhibited the replications of the TK deficient (TK-) mutants of HSV-1 and HSV-2 as well as the TK+ parent strains in vitro. The TK- mutants of HSV-1 and HSV-2 (HSV-1 TK- and HSV-2 TK-) were as susceptible to OXT-G as the TK parent strains. However, the TK- mutants were less susceptible to carbocyclic OXT-G than the TK+ parent strains. We demonstrated synergistic inhibition of the replications of HSV-1 and HSV-2 by ACV and OXT-G in combination, additive inhibition of HSV-1 and HSV-2 by ACV and carbocyclic OXT-G in combination, synergistic inhibition of HSV-1 by OXT-G and carbocyclic OXT-G in combination, and additive inhibition of HSV-2 by these two compounds. We investigated the metabolism of ACV and OXT-G in HSV-1 TK(+)-, HSV-1 TK(-)- and mock-infected Vero cells by thin layer chromatography. ACV-triphosphate increased more in HSV-1 TK(+)-infected Vero cells than in HSV-1 TK(-)- and mock-infected Vero cells. The metabolism of OXT-G had almost the same pattern in HSV-1 TK(+)-, HSV-1 TK(-)- and mock-infected Vero cells. These results suggest that ACV is phosphorylated by virus-induced TK, and OXT-G is phosphorylated by cellular nucleoside and nucleotide kinases.


Prevention of the acute neurotoxic effects of phenytoin on rat peripheral nerve by H7, an inhibitor of protein kinase C. The neurotoxic effects of a single dose of phenytoin (150 mg/kg body weight) alone or 30 min after H7 (a protein kinase C inhibitor) injection (20 mg/kg body weight) were investigated in terms of peripheral neuromuscular function and Na+,K(+)-ATPase activity of the sciatic nerve. This intraperitoneal injection of phenytoin induced complete blockade of muscle action potentials in the dorsal segmental muscles of the rat tail evoked by electric stimulation of the caudal nerve and a 40% decrease in the Na+,K(+)-ATPase activity of the rat sciatic nerve when compared with control values, measured as the difference between total and ouabain-insensitive ATPase activity. Prior administration of H7 resulted in the complete prevention of both effects. Implications of protein kinase C inhibition in phenytoin neurotoxicity are discussed.


Esterase activity in human breast cyst fluid: associations with steroid sulfates and cations. Human breast cyst fluid (BCF) contains an esterase that on the basis of electrophoretic mobility and response to inhibitors differs from those found in the plasma. From a total of 384 BCF samples analyzed for esterase using p-nitrophenyl hexanoate as substrate, 149 (39%) showed significant activity. The samples had been analyzed for the concentrations of the sulfates of estrone, estriol, dehydroepiandrosterone, as well as the potassium and sodium cations (K+/Na+). The data were submitted to statistical analysis using the Spearman rank order test. The esterase-positive samples exhibited a significant positive association with each of the steroid sulfates and the K+/Na+ ratios. Except for protein concentration, there was no significant correlation between the esterase-positive and esterase-negative cysts. These observations may have physiological significance in that high K+/Na+ ratio cysts have been related to the histological status of the cyst.


Gas chromatography/mass spectrometry of catechol estrogens. Catecholestrogens (CCEs), namely 2- or 4-hydroxyestradiol and hydroxyestrone, are highly polar, reactive, and extremely labile estrogen metabolites in many experimental conditions. For these reasons, indirect assay methods mainly have been used. Some experimental evidence suggests that CCEs are synthesized and biologically active mostly in target cells. At this level, unfortunately, the indirect assays cannot be used. We present a method of gas chromatographic/mass spectral (GC/MS) analysis for the identification of individual CCEs; the major fragmentation ions of authentic estrogen standards as trimethylsilylether derivatives, and the MS patterns of the major CCEs, namely, 2-hydroxyestradiol and hydroxyestrone, are included. Few examples of CCEs detected in human breast cancer tissues and in breast cyst fluids are reported. Sample extracts were submitted to reversed-phase, high-performance liquid chromatography (RP-HPLC) and were quantified by "on line" electrochemical (EC) detection; thereafter, either crude extracts or single eluted peaks were submitted to GC/MS, by which detection limits of less than 5 pmol were attained. As expected, the molecular ion was the most relevant molecule in all but one case. On the contrary, the other relative intensities of major fragmentation ions M -15, M -30, M -90, and M -15 + (-90) were unevenly distributed, although represented in the majority of cases. In all cases, the GC/MS of peak fractions, purified by RP-HPLC and UV detection, confirmed the results of liquid chromatographic analysis combined with EC detection. In contrast, GC/MS of crude extracts was not equally satisfactory. Comparison of a liquid chromatography system with EC detection and the GC/MS approach revealed some inconsistency in quantitation of individual CCEs.(ABSTRACT TRUNCATED AT 250 WORDS)


Synthesis and biological activities of 8(14)a-homocalcitriol. 8(14)a-Homocalcitriol was synthesized and tested for its biologic activities. It exhibited a vitamin D agonist activity profile. The compound was bound to the pig intestinal receptor with an affinity slightly less than calcitriol, showed the same potency in inducing HL 60 cell differentiation and inhibition of keratinocyte proliferation as calcitriol, and was found to be approximately 10-fold less potent in inducing hypercalcemia and hypercalciuria after a single injection in normal rats.


Receptor for sex steroid-binding protein of endometrium membranes: solubilization, partial characterization, and role of estradiol in steroid-binding protein-soluble receptor interaction. The sex steroid-binding protein (SBP) receptor was solubilized from the membranes of human premenopausal endometrium with the zwitterionic detergent CHAPS. The binding activity of the soluble receptor was studied, allowing it to interact with [125I]SBP and precipitating the complex with polyethylene glycol 8,000. The interaction of SBP with the soluble receptor was specific, saturable, and at high affinity. Indeed, the specific binding was definitely improved on the solubilized form of the receptor. The effect exerted by sex steroids on the interaction of SBP with receptor was also examined on both the soluble and membrane-bound forms. At physiologic doses (10(-8) M) estradiol inhibits the binding at a significant extent on the soluble receptor, but not on membrane-bound form. The dose of estradiol required to significantly inhibit the SBP-specific binding was dependent on the form of receptor. In membrane-bound receptor the inhibiting dose of estradiol was higher than its physiologic concentration. Thus, it is likely that, while soluble receptor cannot recognize the complex steroid-SBP, membrane-bound receptor can interact both with "unliganded" SBP and with the estradiol-SBP complex (but not with androgen-SBP complexes) in an estrogen-dependent tissue like human endometrium.


Calcium and inositol 1,4,5-trisphosphate receptors: a complex relationship. Increases in intracellular free Ca2+ concentration ([Ca2+]i), whether initiated by changes in plasma membrane potential or receptor-stimulated polyphosphoinositide hydrolysis, can be astonishingly complex, often occurring as repetitive Ca2+ spikes and regenerative Ca2+ waves that propagate through the cell and sometimes into neighbouring cells. The key to understanding these complex Ca2+ signals lies in understanding the interactions between the different pools from which Ca2+ can rapidly enter the cytosol and the activities of the various Ca(2+)-transporting systems that reverse the process.


Signal integration at the level of protein kinases, protein phosphatases and their substrates. Protein phosphorylation and dephosphorylation is one of the major mechanisms of signal integration in eukaryotic cells. It can be achieved through the phosphorylation of proteins that control the levels of second messengers, through the phosphorylation of protein kinases and phosphatases themselves, or through the reversible phosphorylation of their substrates. This article also highlights the important role of 'multi-site phosphorylation' in signal integration, in which different protein kinases produce additive, synergistic and antagonistic effects on activity by phosphorylating distinct Ser or Thr residues in a single protein.


Multiplicity generates diversity in the retinoic acid signalling pathways. Complexity in the retinoid signalling system arises from a combination of several forms of retinoic acid, multiple cytoplasmic binding proteins and nuclear receptors, and the existence of polymorphic retinoic acid response elements. Additional diversity appears to be generated by heterodimeric interactions between two families of nuclear retinoic acid receptors, and between nuclear retinoic acid receptors and other members of the nuclear receptor superfamily. Thus, a complex array of combinatorial effects is beginning to emerge which may account for the pleiotropic effects of retinoids.


Selective placement of electron spin resonance spin labels: new structural methods for peptides and proteins. Electron spin resonance (ESR) is more powerful than ever as a technique for solving biochemical and biophysical problems. Part of the great utility of ESR arises from the use of modern biochemical methods to place spin labels at important positions along the primary sequence of a peptide or protein.


[The detection of albumin intraperitoneally administered to rabbits in the subclavian lymph node]. With light and electron microscopy, the localization of human albumin labeled with colloidal gold is described in the subclavia lymph nodes of rabbits following an intraperitoneal injection of this labeled albumin. Most of the particles were found in the reticular cells of the sinus, and some particles were identified in the sinus macrophages. No particles were found inside lymph node follicules within 1 hour after injection. All stages of internalization of foreign protein inside lymph node cells were demonstrated.


[The effect of gamma irradiation on the structure and enzymatic activity of the nuclear membrane of the liver in pregnant rats and their embryos]. Morphological and biochemical investigations of pregnant rats and embryo liver cell nuclei after in vivo irradiation in the doses of 1 and 2 Gr revealed their high radiosensitivity at all stages of gestation and embryonal development. At damaging effect of radiation, we managed to observe sharp accumulation of products of lipid peroxide oxidation and suppression of the activities of such enzymes as cytochrome-c-oxidase, NAD.N-cytochrome-c-reductase, ATPase and RNAase in liver nuclei of pregnant rats and embryos. The changes of such a kind are shown to intensify with the increasing of irradiation doses. The most profound inhibition of the activities of these enzymes in liver nuclei of embryos irradiated in utero was observed during the period of organogenesis (the 13th day of the development) and in fetal period of embryogenesis (the 17th day of the development), as well as at the 13th and 17th day of gestation. The morphological data also demonstrate the high level of cell nucleus sensitivity to the action of radiation during gestation and embryogenesis.


[A comparative ultracytochemical and biochemical study of the ATPases of the choroid plexus in aging]. A complex study was undertaken on the ATPase and an ouabain-sensitive potassium-dependent p-nitrophenylphosphatase part of the Na, K-ATPase complex at the ultrastructural level, in addition to a biochemical assay of the total ATPase activity and activity of Na, K-ATPase of the choroid plexus (CP) of brain ventricles in adult (6-8 mo) and old (26-28 mo) rats. A correlation between the results of cytochemical and biochemical analyses was noted pointing to an age-related decrease in ATPase activity of the CP. Especially marked was the decrease in Na, K-ATPase activity that evidenced for the reduced level of liquor production in the CP during aging. Identified were the sites of a predominant localization of enzymes on the plasma membranes and intracellular organelles of the CP epitheliocytes. The latter fact was associated with their involvement in the processes of energy transformation and transport of substances. The data of the ultracytochemical and biochemical analyses, together with the results of the ultrastructural investigation, indicate an age-related decrease in the functional activity of the CP that represents one of the essential links in the mechanism of brain aging.


Acute and chronic faucitis of domestic cats. A feline calicivirus-induced disease. The lesions of acute feline calicivirus infection are of a transient vesiculo-ulcerative nature and involve, to varying degrees, the palate, tongue, gingiva, lips, nasal philtrum, and oral fauces. Chronic ulceroproliferative faucitis is a specific but uncommon sequelae to persistent feline calicivirus oral carriage.


Cross-reactivity of existing equine influenza vaccines with a new strain of equine influenza virus from China. A novel strain of equine influenza virus, influenza A/equine/Jilin (China)/1/89, has emerged which is genetically distinct from all earlier strains of equine influenza. It is therefore possible that the vaccines against equine influenza may be unable to protect horses against disease caused by this virus strain. In vitro serological assays established that there were low levels of immunological cross-reactivity between the new virus, the current vaccine strains and the strains of equine-2 influenza virus now in circulation.


Evaluation of two ELISA kits for the detection of Aujeszky's disease antibodies in pigs. This work describes the evaluation of two commercial ELISA kits for the detection of gI antibodies against Aujeszky's disease. A collection of experimental sera from infected pigs, field sample sera, and sera from pigs vaccinated with seven different modified gI-negative commercial vaccines were used to evaluate each test. Both ELISA kits showed good reproducibility, and specificity, but differences could be appreciated in sensitivity when sera obtained at early stages of infection was analysed. These results also indicated that both kits could be used in conjunction with the seven vaccines evaluated in this study.


Isolation of a virus serologically related to the bluetongue group from an outbreak of haemorrhagic disease among exotic deer in Saudi Arabia. In February 1991 a severe haemorrhagic disease affected exotic deer aged over six months in the Al-Hofuf area of the eastern region of Saudi Arabia. The morbidity rate was 40 per cent and the case fatality rate was 60 per cent. The clinical signs were high temperature (up to 41.5 degrees C), nasal discharge, slight salivation and lacrimation, congestion of the conjunctivae, torticollis, tremors when trying to stand, recumbency and coma leading to death. Post mortem examination revealed a severe haemorrhagic disease. A virus, serologically related to the bluetongue serogroup, was isolated from the deer. Sheep and goats kept on the same farm did not show any clinical signs. The epidemiology of the outbreak is discussed.


Serological responses of specific pathogen-free foals to equine herpesvirus-1: primary and secondary infection, and reactivation. Serum antibody (virus neutralisation, complement fixation, IgM and IgG) responses to equine herpesvirus-1 (EHV-1) infection were measured in six foals which were initially free from EHV-1 and EHV-4 infection and maternally-derived antibodies. Following primary infection, high titres of virus neutralisation and complement fixation antibodies were detectable against EHV-1, however, corresponding antibody levels against EHV-4 were low or inapparent, although the two viruses share a number of cross-reactive epitopes. In addition, following the primary infection with EHV-1, IgM levels increased before those of IgG, virus neutralisation and complement fixation antibodies, peaked sooner and thereafter declined. Stimulation of IgM levels was observed on secondary infection with EHV-1 given 61 days later. In contrast, IgG, virus neutralisation and complement fixation antibodies following primary infection were more sustained and no increase in their levels was observed on secondary infection. No consistent changes in IgM or IgG levels were seen after administration of dexamethasone to reactivate latent virus.


Production and characterization of monoclonal antibodies to cervine herpesvirus-1. Fourteen hybridoma cell lines secreting monoclonal antibodies (Mab) to cervine herpesvirus-1 (CerHV-1) produced following the fusion of NSO myeloma cells with splenocytes of BALB/c mice previously immunized with gradient purified CerHV-1 were selected using an indirect enzyme-linked immunosorbent assay (ELISA) employing CerHV-1 antigen and tested by the ELISA against four other ruminant alphaherpesviruses from cattle (bovine herpesvirus type 1.1 and 1.2) goat (caprine herpesvirus-2) and reindeer (rangiferine herpesvirus-1). Comparison of all five ruminant alphaherpesviruses with these Mabs confirmed their close antigenic relationships, with two Mabs reacting against all viruses. Ten Mabs which were able to differentiate between the viruses reacted with a 64 kDa polypeptide in a western blot. Four Mabs including two specific only for CerHV-1 with neutralizing activity against the virus used for immunization were directed against a 74 kDa viral protein.


Isolation of bluetongue and epizootic hemorrhagic disease viruses from mosquitoes collected in Indonesia. Three viruses isolated from anopheline mosquitoes in Indonesia have been identified as bluetongue and epizootic hemorrhagic disease viruses. Another virus isolate showed no relationship to other orbiviruses tested and should be regarded as a new virus; the name Golok is proposed for it. The mosquitoes were collected in 1980 and 1981 in a program designed to isolate flaviviruses infecting humans. It is apparent that such collections of arthropods which feed on large mammals could be screened for other viruses which may infect domestic livestock.


Foot-and-mouth disease: detection of antibodies in cattle sera by blocking ELISA. A blocking ELISA was developed for the detection of antibodies to foot-and-mouth disease virus SAT1, SAT2 and SAT3 and for the quantification of antibodies on a single dilution of serum. The avidin-biotin system was used. The test was compared with the liquid-phase ELISA executed at the World Reference Laboratory for foot-and-mouth disease. It was found to have favourable logistics and combined high specificity with high sensitivity. The quantitative test using a single dilution of serum was resource saving and proved to be a reliable and precise method for the assessment of antibody levels.


Recombinant polypeptide from the gp48 region of the bovine viral diarrhea virus (BVDV) detects serum antibodies in vaccinated and infected cattle. To characterize the immune response of cattle to bovine viral diarrhea virus (BVDV) glycoprotein gp48, we have produced a large amount of recombinant glutathione-s-transferase-gp48 (GST-gp48) fusion protein in Escherichia coli. Antibodies to gp48 were present in cattle vaccinated with killed or modified-live virus vaccination, or following natural infection. These results were in agreement with results of serum neutralization (SN) test which detected gp53 of BVDV.


Comparison of antibody and cell-mediated immune responses in horses following feeding of a novel dietary antigen, ovalbumin, and rotavirus. Adult ponies which were fed ovalbumin (OVA) daily for 2 weeks had significantly greater serum anti-OVA IgG (P = 0.001) and antigen specific lymphocyte responses (P = 0.031) after intramuscular injection with OVA given with saponin than control ponies which had not been fed the antigen. This suggests that, despite the lack of evidence of B- or T-cell activation in peripheral blood during the period of OVA feeding, the animals were primed for an active secondary immune response. Adult ponies were challenged with equine rotavirus, strain H-2, but no statistically significant differences were found in serum IgG-associated antibody responses or antigen-specific lymphocyte responses between the rotavirus-challenged group and the control group, either following rotavirus challenge or intramuscular injection of rotavirus antigen given with saponin. Our findings, that feeding the non-replicating protein antigen OVA appeared to prime for an increased immune response rather than inducing oral tolerance, may be of relevance to future studies on the way the equine gastrointestinal tract handles usually harmless antigens.


Immune dysfunction following infection with chicken anemia agent and infectious bursal disease virus. I. Kinetic alterations of avian lymphocyte subpopulations. The potential effect of chicken anemia agent (CAA) alone or in combination with infectious bursal disease virus (IBDV) on the immune system of young chickens was determined by measuring alterations in hematocrit values, lymphoid organ-to-body weight ratios and lymphoid cell concentrations at 4, 7, 10, 14, 17, 21, 28 and 42 days post-inoculation (PI). Lymphocyte subpopulations were identified and counted by flow cytometry using cell suspensions stained with monoclonal antibodies (Mabs) for panlymphocytes (K55), cytotoxic T-cells (CTLA3), T-helper cells (CT3), Ia-expressing cells (P2M11) and macrophages (P7). Chicken anemia agent induced a substantial but transient decrease in hematocrit value, thymus-to-body weight ratio and bursa-to-body weight ratio between 7 and 21 days PI corresponding to a generalized lymphocytopenia in the thymus, bursa and spleen. However, cytotoxic T-cell, T-helper cell and Ia-expressing cell concentrations increased in the bone marrow of birds inoculated with CAA alone or in combination with IBDV during the same time period. T-helper-to-cytotoxic T-cell ratios increased in the thymus and spleen during severe lymphocytopenia, indicating a selective decrease in cytotoxic T-cells. T-helper-to-cytotoxic T-cells ratios increased in the bone marrow, indicating a selective increase in T-helper cell concentrations. The increase in Ia-expressing cells in the bone marrow may be a reflection of increased number of activated T-cells which express Ia antigen. Infectious bursal disease virus alone induced a persistent depression of Ia-expressing cells in the bursa and the spleen and no measurable change in the bone marrow lymphocyte subpopulations. Chickens inoculated simultaneously with CAA and IBDV experienced clinical signs observed in chickens inoculated with each virus separately with a prolonged acute phase prior to recovery or mortality.


Immune dysfunction following infection with chicken anemia agent and infectious bursal disease virus. II. Alterations of in vitro lymphoproliferation and in vivo immune responses. To determine the functional impact of alterations in lymphocyte concentrations and ratios following infection with chicken anemia agent (CAA) alone or in combination with infectious bursal disease virus (IBDV) on the immune system of young chickens, in vitro lymphoproliferation assays and in vivo responses to vaccination with several common viral agents were assessed at various time intervals post-inoculation (PI). Concanavalin A (Con A), phytohemagglutinin (PHA) and pokeweed mitogen (PWM) stimulation of splenic lymphocytes (SPL) collected from control birds could not be detected until 10-14 days PI. Infection with CAA was characterized by significantly higher PWM stimulation of SPL at 17 days PI and significantly lower PWM stimulation of peripheral blood lymphocytes (PBL) at 14 days PI, compared with uninfected controls. Concanavalin A and PWM stimulation of SPL was significantly increased in birds inoculated with IBDV alone. Lymphocytes harvested from birds inoculated simultaneously with CAA and IBDV had significantly lower responses. Effects on humoral and cell-mediated immunity following CAA and/or IBDV were determined by evaluating vaccination responses to Newcastle disease virus (NDV), fowl pox virus (FPV) and infectious laryngotracheitis virus (ILTV) during the acute phase of CAA infection (2 weeks PI). Vaccination of birds 2 weeks following CAA infection at 1 day of age resulted in decreased protection against NDV (85.7%) and ILTV (7.1%) challenge compared with protection rates in control birds (100% and 53.3% respectively). Infectious bursal disease virus infection was associated with decreased protection against NDV (60%) only. Concomitant infection at 1 day of age resulted in a greater reduction in NDV challenge protection (33.3%), slightly decreased FPV protection (87.5%), increased numbers of persistent FPV vaccination lesions and increased protection against ILTV challenge (71.4%). Vaccination of birds 2 weeks following CAA infection at 2 weeks of age resulted in slightly decreased NDV humoral antibody, development of persistent FPV vaccination lesions (17%) and increased immunity to ILTV challenge compared with control birds (83.3% vs. 66.7%). Chickens inoculated with IBDV alone displayed a more severe depression in NDV antibody titers and only a slight decrease in ILTV protection. Vaccination following concomitant infection at 2 weeks of age resulted in a higher percentage of FPV persistent vaccination lesions (39%) and greatly enhanced immunity to ILTV challenge (100%).


Low p53 protein expression in salivary gland tumours compared with lung carcinomas. Fifty-one salivary gland tumours (23 pleomorphic adenomas, 5 Warthin's tumours, 12 mucoepidermoid carcinomas, 7 adenoid cystic carcinomas, 3 undifferentiated carcinomas and 1 acinic cell tumour) and 27 lung carcinomas (18 squamous cell carcinomas) were analysed immunohistochemically for the expression of p53 nuclear phosphoprotein. Eight out of 51 (16%) salivary gland tumours were p53 positive. Three of these were benign and 5 malignant. All 3 benign salivary gland tumours were pleomorphic adenomas and expressed only occasional nuclear positivity with less than 1% of tumour cells positive. Of the 5 p53-positive malignant tumours, 3 were mucoepidermoid carcinomas and 2 undifferentiated carcinomas. The malignant salivary gland tumours expressed more than 1% of positive nuclei in every case. Seventeen lung carcinomas were p53 positive (63%). Thirteen of these were squamous cell carcinomas, 3 were adenocarcinomas and 1 small cell lung carcinoma. The results show that mutations of the p53 gene may be infrequent in salivary gland tumours when compared with lung carcinomas. The relatively indolent course of some histological types of malignant salivary gland tumours could be associated with the preservation of the non-mutated p53 gene in most of these tumours. The presence of p53 positivity in some pleomorphic adenomas might, on one hand, suggest that p53 gene alterations are also present in these tumours; on the other hand, the accumulation of the p53 protein in these tumours might also be due to some unknown mechanism, not necessarily related to p53 gene mutation.


Enteropathy-associated T-cell lymphoma in a renal transplant patient with evidence of Epstein-Barr virus involvement. The clinical and histological findings in a 54-year-old patient with enteropathy-associated T-cell lymphoma (EATL) occurring 18 years after renal transplantation are presented. Ten years after adult-onset coeliac disease the patient developed medium to large T-cell non-Hodgkin's lymphoma of the small intestine. Epstein-Barr virus (EBV) genome was detected by polymerase chain reaction in the lymphoma tissue and localized via Epstein-Barr virus RNAs in situ hybridization to some of the tumour cells. This is the first case report of EBV-positive EATL occurring in the setting of immunosuppression.


[The interrelationship between humoral factors and the indices of left ventricular myocardial hypertrophy in hypertension]. Left ventricular myocardium hypertrophy, adrenocorticotropin, cortisol, renin and aldosterone were evaluated in 100 patients subdivided in three groups depending on the type of central hypertensive disease of stage two hemodynamics. Their correlation coefficients were assessed before and after treatment. It was found that patients with hypertensive disease (stage II) did not show any regularities in the correlation between the content of hormones in the blood and signs of left ventricular myocardium hypertrophy. Hence, aldosterone, renin, ACTH, cortisol do not essentially influence the development of left ventricular myocardium hypertrophy.


Renal transplant and cytomegalovirus infection: experience in a community hospital program. Cytomegalovirus (CMV) is the most frequently occurring opportunistic pathogen and single most important infectious agent in renal transplant recipients, contributing significantly to their morbidity and mortality. Infection generally occurs in the first four months after transplant and develops in about two-thirds of these patients overall. Cytomegalovirus infection may occur as a primary disease, a reactivation, or as a superinfection and may encompass a spectrum of diseases ranging from asymptomatic to fatal. This study reviews CMV donor and recipient status, prophylaxis, rejection episodes, immunosuppression and subsequent development of disease in 100 transplant cases in 98 patients which were performed at the charleston Area Medical Center between September 1987 and August 1991, and contrasts these findings to those from larger centers. Sixty-five recipients were positive for cytomegalovirus serologically at the time of transplantation, and of the 35 patients who were CMV negative, 20 received a kidney from a CMV-positive donor and a total of six patients have developed symptomatic disease to date.


[A case of idiopathic hypercalcemia (hypersensitivity to vitamin D 3]. Hypercalcaemia occurs in two forms: mild and severe. In the mild form, usually in young infants the characteristic signs of the severe from (Williams syndrome) are absent, and thus it may cause diagnostic difficulties. Because of that, in infants with muscular hypotonia, growth arrest, constipation and apathy the possibility of idiopathic hypercalcaemia, apart from rickets, should be considered.


Direct injection/h.p.l.c. methods for the analysis of drugs in biological samples. 1. Direct injection h.p.l.c. methods for zaprinast, and pantoprazole and its sulphone metabolite were developed. 2. Optimal recovery of pantoprazole and its sulphone metabolite was effected by the absence of transfer losses and the effective adjustment of sample pH on-line. 3. Acetonitrile reduced the recovery of pantoprazole and its sulphone metabolite at acetonitrile concentrations greater than 5% in serum. 4. Direct injection h.p.l.c. methods minimize sample handling losses, reduce human contact with biological samples and are sufficiently accurate and reproducible to be used to support pharmacodynamic and toxicokinetic studies.


Active protection of mice against Salmonella typhi by immunization with strain-specific porins. NIH mice were immunized with between 2.5 and 30 micrograms of two highly purified porins, 34 kDa and 36 kDa, isolated from the virulent strain Salmonella typhi 9,12, Vi:d. Of mice immunized with 10 micrograms of porins, 90% were protected against a challenge with up to 500 LD50 (50% lethal doses) of S. typhi 9,12,Vi:d and only 30% protection was observed in mice immunized with the same dose of porins but challenged with the heterologous strain Salmonella typhimurium. These results demonstrate the utility of porins for the induction of a protective status against S. typhi in mice.


Immunogenicity of inactivated purified tissue culture vaccine against hepatitis A (HepAvac) assessed in laboratory rodents. Immune response of laboratory rodents (guinea-pigs, CBA and Balb/c mice, Wistar and August rats) to inactivated hepatitis A vaccine was quantitatively assessed. Under comparable conditions of experiment, the mice showed the highest antibody titres and were capable of reacting to the lower doses of immunogen; meanwhile their individual variations in immune response were more pronounced; white rats were the least susceptible to the vaccine, demonstrating the minimal antibody formation; guinea-pigs produced antibody at intermediate levels but the antibody titres were the most homogeneous. The enhancing effect of aluminium hydroxide was observed in guinea-pigs examined at the late postimmunization stage. Differences in immunogenicity of three vaccine lots were essentially similar when these lots were tested as undiluted preparations in guinea-pigs and mice for mean antibody titres and in mice for 50% immune response using serial dilutions of vaccine. All three tests could be routinely employed for vaccine immunogenicity control.


In vivo model for evaluation of species-specific virus vaccines. It is difficult to evaluate the protective efficacy of species-specific viruses of humans and expensive companion animals where there is no non-human animal model. This study describes an in vivo model system which allows simultaneous operation of humoral, cell-mediated, interferon-like or other unidentified immunological defence mechanisms. There was evidence of in vivo inactivation of both enveloped and unenveloped DNA and RNA viruses including retrovirus mouse sarcoma virus/mouse leukaemia virus as evaluated by assay of the enzyme reverse transcriptase. This model will allow examination of vaccine efficacy in immunocompetent host animals while avoiding morbidity and/or mortality from virus infection in these animals.


Liver resection for hepatocellular carcinoma (HCC) with direct removal of tumor thrombi in the main portal vein. Since the tumor thrombus in the main portal vein appears in the terminal stage of hepatocellular carcinoma (HCC), any attempt to remove it surgically is thought to be impractical as the malignancy itself cannot be entirely removed. During the past 5 years, we have performed tumor thrombectomy combined with hepatectomy in 29 of 298 patients with HCC. This combined therapy was initially decided upon as an emergency measure to prevent impending rupture of esophageal varices, rather than to improve patient survival. Since portal flow was obtained after removal of thrombi, this condition enabled transcatheter arterial embolization (TAE) and/or percutaneous ethanol injection therapy (PEIT). Although improved patient survival was not the primary goal of the emergency operation and there was an operative mortality of 11%, half of the other patients in the present series had unexpectedly high survival rates of 1 year (52.2%), 2 years (23.2%), and 3 years (11.6%), which were significantly higher than in patients not undergoing operation (n = 22).


Seroimmunity to poliomyelitis in an Albanian immigrant population. The immunity against poliomyelitis in a representative sample of the Albanian population recently immigrant to Italy was evaluated. A significant number of the subjects examined lacked protective antibodies against one or more polioviruses. The most prevalent seronegativity related to poliovirus type 3 (41.3%), followed by poliovirus type 1 (21.5%). This result was more consistent in the younger age groups. Our data suggest the hypothesis that this problem arises from the use of a vaccine of discontinuous efficacy caused by defective preservation. A poliovaccine booster dose for all emigree subjects aged less than 15 years is suggested.


Diabetes mellitus in cattle infected with bovine viral diarrhea mucosal disease virus. Four Japanese black cattle and four Holstein-Friesian cows were diagnosed with diabetes mellitus. Based on intravenous glucose tolerance tests, all these animals were believed to be insulin-dependent diabetics. Moreover, bovine viral diarrhea-mucosal disease (BVD-MD) virus was isolated and mucosal lesions and diarrhea were recognized in all cases. There was no genetic relation among the affected cattle. The breeding places of these cattle were different to each other. Therefore, it is suggested that BVD-MD virus induced the diabetes mellitus.


Antigens involved in vaccination of swine against Aujeszky's disease (pseudorabies) virus. The polypeptide and glycopolypeptide composition of a local virulent Aujeszky's disease virus (suid herpesvirus 1, SHV-1) strain (E-974) was determined in order to characterize the individual SHV-1 antigens inducing the serological responses in immunized and non-immunized animals. A commercially available inactivated vaccine of known efficacy and three experimental immunogen preparations (whole inactivated SHV-1 particles, lectin-purified glycoproteins from SHV-1 culture, and a combination of both) were used for immunization. Sera of two-month old immunized and non-immunized animals were analyzed by ELISA, seroneutralization and Western immunoblotting prior to and following challenge with E-974. Sera of 7- to 30-day-old piglets littered by immunized and non-immunized sows were likewise analyzed by immunoblotting. The following variables were determined: the total level of anti-SHV-1 antibodies, the level of neutralizing antibodies, the IgG responses to individual SHV-1 antigens, and the clinical parameters and degree of protection of the animals. The whole-particle experimental immunogen conferred greatest protection, but correlation between antibody levels and the degree of protection was imperfect. Serological responses seemed to be directed against certain structural polypeptides and viral envelope glycoproteins. The glycoprotein immunogen caused a selective response to bands which closely resemble the glycopolypeptides gII and gIII. A 71 kDa component of uncertain location within the viral structure appeared to be one of the main antigens involved in porcine serological response to SHV-1 and colostral protection of piglets.


Repeat operation for nodular recurrent hepatocellular carcinoma within the cirrhotic liver remnant: a comparison with transcatheter arterial chemoembolization. Management of recurrent hepatocellular carcinoma in a cirrhotic liver remnant is a difficult but challenging problem. To investigate the difference in survival between treatment by repeat resection and treatment by transcatheter arterial chemoembolization (TAE), a retrospective controlled study was conducted. Four patients with nodular recurrence received limited second operations which included right hepatic segmentectomy (2 patients), left lateral segmentectomy (1 patient), and subsegmental wedge resection (1 patient). Eight matched patients received a total of 16 repeated sessions of chemoembolizations. Complications of the TAE group consisted of gastrointestinal bleeding (2 patients), acute pancreatitis (1 patient), and acute cholecystitis (1 patient). No complication developed in the resection group. The 4 patients undergoing a second operation have survived 21, 26, 34, and 54 months after repeat surgery. Seven (87.5%) of the 8 patients receiving TAE died 4 to 11 months after TAE. The resection group survived significantly longer than the TAE group (p < 0.01). Our results suggest that it is more advisable to perform a second operation than to undertake chemoembolizations for patients with cirrhosis and nodular recurrent hepatocellular carcinoma with acceptable functional liver reserve.


[The rapid differential diagnosis of bacterial and viral meningitis by using the lysozyme test]. The authors have modified the technique of the lysozyme test by adding polimixin M sulfate into the gel bacterial medium. Rapid diagnosis with the use of this test is based on different time of the appearance of the lysis areas: in bacterial meningitides the CSF lysozyme activity is detectable within 15-120 min, whereas in viral meningitides it manifests 40-50 min later or does not manifest at all. The results were found to depend on the time of the CSF collection: the earlier the CSF samples were obtained, the higher was the share of positive results.


[Clinical polymorphism and genetic heterogeneity of hereditary spastic ataxia]. The authors systematized the descriptions of different ataxic degenerative syndromes associated with the development of pyramidal symptomatology. Demonstrated the genetic pleomorphism of hereditary spastic ataxias, described the clinical features characteristic of different types of inheritance of spastic ataxias. Based on the authors' observations and analysis of the reported data the following criteria for the diagnosis "spastic ataxia" are suggested: autosomal dominant inheritance type, with the onset on the 3rd-4th decade of life, cerebellar ataxia and dysarthria coupled with tendinous hyperreflexia and the rise of the muscular tone by the spastic type.


[Cerebrotendinous xanthomatosis].

[Cerebrotendinous xanthomatosis]. Cerebrotendinous xanthomatosis (CTX) is a rare familial lipid storage disease caused by defective bile acid synthesis. As a result, cholestanol, a derivative of cholesterol, is accumulated by virtually every tissue, predominantly by the nervous system, xanthomas and bile. Clinically, progressive neurologic dysfunction, tendon xanthomas, cataracts, osteoporosis and atherosclerosis are commonly found. Replacement therapy with chenodeoxycholic acid (750 mg/day), a primary bile acid, which is almost absent from the bile in CTX, reduces elevated cholestanol synthesis and concentrations and improves neurologic function in this disease.