A myofibrillar protein phosphatase from rabbit skeletal muscle contains the beta isoform of protein phosphatase-1 complexed to a regulatory subunit which greatly enhances the dephosphorylation of myosin. A form of protein phosphatase-1 (PP1M), which possesses 25-fold higher activity towards the P light chain of myosin (in heavy meromyosin) than other forms of protein phosphatase-1, was purified over 200,000-fold from the myofibrillar fraction of rabbit skeletal muscle. PP1M, which eluted from Superose 12 with an apparent molecular mass of 60 kDa, was dissociated by LiBr into two subunits. One of these displayed enzymic properties identical to those of the catalytic subunit of protein phosphatase-1 (PP1C) and was identified as the beta isoform of PP1C by amino acid sequencing. The second subunit had no intrinsic protein phosphatase activity, but greatly increased the rate at which PP1C dephosphorylated skeletal-muscle heavy meromyosin and decreased the rate at which it dephosphorylated glycogen phosphorylase. The properties of PP1M, together with those of smooth muscle PP1M [Alessi, D., MacDougall, L. K., Sola, M. M., Ikebe, M. & Cohen, P. (1992) Eur. J. Biochem. 210, 1023-1035] and the previously characterised glycogen-associated form of protein phosphatase-1 (PP1G), indicate that the subcellular localisation and substrate specificity of PP1 is determined by its interaction with specific targetting subunits.


Hydration of the Gly2 and Gly3 peptide oxygens of [Leu5]-enkephalin in aqueous solution as revealed by the combined use of 17O-NMR and Fourier-transform infrared spectroscopy. Solvent-induced and temperature-induced 17O chemical shifts of [17O-Gly2, Leu5]-enkephalin and [17O-Gly3, Leu5]-enkephalin and solvent-induced spectral modifications of the amide-I' stretching vibrations of [1-13C-Gly2, Leu5]-enkephalin and [1-13C-Gly2, Leu5]-enkephalin are reported and correlated with the spectroscopic characteristics of model amides. It is demonstrated that both Gly2 and Gly3 peptide oxygens are motionally equivalent and form solvation species which are essentially monohydrated in aqueous solution, contrary to several simple amides and model peptides in which water largely forms dihydrates. It is shown that the combined use of 17O-NMR and Fourier transform infrared is a unique methodology for studying the hydration state of specific peptide oxygens in peptide hormones.


Characterization of a biologically active extracellular domain of fibroblast growth factor receptor 1 expressed in Escherichia coli. The functional features of a recombinant fibroblast growth factor (FGF) receptor (FGF-R) were investigated by expressing at high level in Escherichia coli a soluble non-glycosylated form of FGF-R1. The extracellular domain of the mature protein (XC-FGF-R), comprising the first 356 amino acids, was purified from a large-scale fermentation. After cell lysis, the protein was quantitatively found in the pellet. XC-FGF-R was solubilized using guanidine/HCl and allowed to refold using two dialysis steps. The refolded protein was obtained in a homogeneous form after ammonium sulphate precipitation and gel-filtration chromatography. The soluble receptor had the ability to form a complex with recombinant human basic FGF (rhbFGF) in solution, as demonstrated by immunoprecipitation with anti-(FGF-R) serum. Formation of a rhbFGF/XC-FGF-R complex was visualized by cross-linking experiments. Quantitative binding experiments with the XC-FGF-R immobilized on Affi-Gel resin showed high binding affinity for 125I-bFGF (Kd = 5-10 nM). Purified XC-FGF-R inhibited binding of 125I-bFGF to its high-affinity receptors on baby hamster kidney cells. These data suggest that glycosylation of the FGF-R is not necessary for its ligand-binding activity. The use of an E. coli expression system resulted in the efficient production of a soluble receptor in a form suitable for ligand/receptor structural studies and screening of new potential agonists and antagonists of angiogenesis. These results indicate that E. coli can be used for the production of complex molecules such as Ig-like receptors.


Purification and characterization of glycerate kinase from a serine-producing methylotroph, Hyphomicrobium methylovorum GM2. The glycerate kinase of a serine-producing methylotroph, Hyphomicrobium methylovorum GM2, was purified to complete homogeneity and characterized, the first time for an enzyme from a methylotroph. The enzyme was a monomer with a molecular mass about 41-52 kDa. The enzyme was stable against heating at 35 degrees C for 30 min at pH values over 6-10. Maximum activity was observed at pH 8.0 and around 50 degrees C. The Km values for D-glycerate and ATP were 0.13 mM and 0.13 mM, respectively. The enzyme showed high specificity for D-glycerate, and was activated by potassium and ammonium ions. The reaction product of the enzyme was identified as 2-phosphoglycerate.


Crystallographic studies of the binding of protonated and unprotonated inhibitors to carbonic anhydrase using hydrogen sulphide and nitrate anions. The structures of human carbonic-anhydrase-II complexes with the anionic inhibitors hydrogen sulphide (HS-) and nitrate (NO3-) have been determined by X-ray diffraction at 0.19-nm resolution from crystals soaked at pH 7.8 and 6.0, respectively. The modes of binding of these two anions differ markedly from each other. The strong inhibitor HS- replaces the native zinc-bound water/hydroxide (Wat263) leaving the tetrahedral metal geometry unaltered and acts as a hydrogen-bonding donor towards Thr199 gamma. The weak NO3- inhibitor does not displace Wat263 from the metal coordination but occupies a fifth binding site changing the zinc coordination polyhedron into a slightly distorted trigonal bipyramid. The interaction of NO3- with the metal is weak; the nearest of its oxygen atoms being at a distance of 0.28 nm from the zinc ion. The binding of nitrate to the enzyme is completed by a hydrogen bond to the metal coordinated Wat263 and a second one to a water molecule of the active-site cavity. The structures of the two complexes help to rationalize the binding of anionic inhibitors to carbonic anhydrase and the binding mode displayed by NO39 may be relevant to the catalytic mechanism.


Structural studies of Desulfovibrio vulgaris ferrocytochrome c3 by two-dimensional NMR. Two-dimensional NMR has been used to make specific assignments for the four haems in Desulfovibrio vulgaris (Hildenborough) ferrocytochrome c3 and to determine their haem core architecture. The NMR signals from the haem protons were assigned according to type using two-dimensional NMR experiments which led to four sets of signals, one for each of the haems. Specific assignments were obtained by calculating the ring current shifts which arise from other haems and aromatic residues. Observation of interhaem NOEs confirmed the assignments and established that the relative orientation of the haems is identical to that found in the crystal structure of D. vulgaris (Miyazaki F.) ferricytochrome c3. Assignments were also made for all the aromatic residues except for the haem ligands and F20, which is shifted under the main envelope of signals. The NOEs observed between these aromatic protons and haem protons confirm the similarity between the structures in solution and in the crystal. The assignments reported here are the basis for the cross-assignments of the four microscopic haem redox potentials to specific haems in the protein structure [Salgueiro, C. A., Turner, D. L., Santos, H., LeGall, J. and Xavier, A. V. (1992) FEBS Lett., in the press]


Overproduction of prismane protein in Desulfovibrio vulgaris (Hildenborough): evidence for a second S = 1/2-spin system in the one-electron reduced state. The gene encoding the prismane protein from Desulfovibrio vulgaris (Hildenborough) was inserted into broad-host-range vector pSUP104. The recombinant plasmid, pJSP104, was transferred to D. vulgaris by conjugal plasmid transfer. In the transconjugant D. vulgaris cells the prismane protein was 25-fold overproduced. The overproduced prismane protein was characterized by molecular mass, isoelectric point, iron content and spectroscopical properties. Both the iron content and the ultraviolet/visible spectrum are identical to the wild-type protein indicating that iron incorporation in the overproduced protein is complete. EPR spectra of the dithionite-reduced form of the overproduced protein indicated that the Fe-S cluster might occur in a similar structure as found in inorganic model compounds containing a [6Fe-6S] prismane core. The as-isolated overproduced protein showed the presence of a second S = 1/2 spin system that was also detected in the corresponding prismane protein from D. desulfuricans (ATCC 27774), but not in the protein from wild-type D. vulgaris. This additional signal was irreversibly transformed to the 'wild-type' high-spin and low-spin systems upon two reduction/re-oxidation cycles. It is shown that the EPR spectroscopy of the overproduced prismane protein is very similar to that of the D. desulfuricans enzyme and, with the exception of the second S = 1/2 spin system, to that of the prismane protein from wild-type D. vulgaris. Contrary to claims for the D. desulfuricans protein, it is shown here that all data can be fully explained assuming a single [6Fe-6S] cluster, that might be titrated into four different redox states and occurs in up to three different spin systems in the one-electron reduced state.


A comparative study of lisinopril and atenolol on low degree urinary albumin excretion, renal function and haemodynamics in uncomplicated, primary hypertension. The presence of slightly increased urinary albumin excretion (UAE), even at levels well below levels detectable by an ordinary dipstick, has been suggested as a predictor of cardiovascular morbidity and as a reflection of the degree of overall vascular permeability. The aim of the present investigation was to study the effects of two different antihypertensive drug regimens, an ACE inhibitor and a beta-adrenoceptor antagonist, on the low UAE rate observed in subjects with uncomplicated, mild to moderate primary hypertension. After a 4-week placebo run-in period, 49 patients (mean age 54 y) were randomly assigned in a double blind manner either to further 4 weeks on placebo (P, n = 15), 8 weeks on lisinopril (L, n = 17; 20 mg/40 mg o.d.) or 8 weeks on atenolol (A, n = 17; 50 mg/100 mg o.d.). The 24-h UAE was measured every second week. At entry and after 4 weeks the glomerular filtration rate and the renal plasma flow were measured. Both drugs lowered blood pressure (BP) to a similar extent after 4 and 8 weeks of treatment; the blood pressures were 160/106 (P), 159/104 (L) and 154/103 (A) at entry, and 133/83 (L) and 134/87 (A) at the end of the study after 8 weeks. On entry the 24-h UAE in all patients ranged from 4 to 49 mg (mean 14.1 mg), and it did not differ significantly between groups.(ABSTRACT TRUNCATED AT 250 WORDS)


The tolerance to and pharmacokinetics of penciclovir (BRL 39,123A), a novel antiherpes agent, administered by intravenous infusion to healthy subjects. The tolerance to and pharmacokinetics of intravenously administered penciclovir (BRL 39,123A), a novel anti-herpes agent, were investigated in 15 healthy male subjects. The volunteers were divided into three groups, receiving either 10, 15 or 20 mg/kg penciclovir by a 60 min constant-rate infusion. Blood samples were taken sequentially up to 48 h after the start of the infusion and urine collections made at appropriate intervals up to 72 h. After a simple solid phase extraction, concentrations of penciclovir in plasma and urine were determined using HPLC with U.V. detection. Mean values of Cmax, corresponding usually with the end of infusion, and of AUC appeared to increase proportionately with dose. Furthermore, there was no evidence that dose significantly affected any individual pharmacokinetic parameter. Penciclovir was distributed into tissues with an overall mean volume of distribution of approximately 1.5 l.kg-1, i.e. approximately double that of body water. It was rapidly eliminated, with a mean total plasma clearance of 39.3 l.h-1, and a mean terminal-phase half-life of 2.0 h. The majority of the dose, approximately 70%, was excreted unchanged in the urine. Mean renal clearance of BRL 39,123 was 28.1 l.h-1, which exceeds normal glomerular filtration rate and approaches renal plasma flow. At all dose-levels, the infusions of penciclovir were well tolerated, with no evidence of drug-related adverse events.


Local generation and action of angiotensin II in dog iris sphincter muscle. Existence of the renin-angiotensin system was pharmacologically investigated in the dog isolated iris sphincter muscle. The sphincter muscle contracted in response to tetradecapeptide, a synthetic renin substrate, angiotensin (ANG) I and ANG II dose-dependently. The contractions induced by these peptides were suppressed by treatment with saralasin, indomethacin and aspirin. Contractile responses to tetradecapeptide and ANG I were also reduced by KRI-1314, a renin inhibitor, and captopril, respectively. ANG II stimulated the release of prostaglandin (PG) F2 alpha from the sphincter muscle. Angiotensin-converting enzyme activity was measurable in the sphincter muscle. Miosis was observed by intracameral injection of ANG I and ANG II into the anterior chamber. These results strongly suggest that angiotensin generating enzymes function in the sphincter muscle and ANG II produced by these enzymes contracts the sphincter muscle via the formation of PG (s), possibly PG F2 alpha.


Endocrine activation mimics the adverse effects of prenatal stress on the neuromotor development of the infant primate. Pregnant female rhesus monkeys were exposed to a 2-week period of adrenocorticotropic hormone (ACTH) to determine whether it would affect the early neuromotor development of their fetuses in a manner similar to that observed after psychological stressors. During the first month after birth, infants were tested on two occasions with a modification of the Brazelton Newborn Behavioral Assessment Scale. Infants derived from ACTH-treated pregnancies showed early impairments in motor coordination and muscle tonicity and shorter attention spans as compared to controls. In addition, on a temperament rating scale, infants from the ACTH condition were more irritable and difficult to console. These findings indicate that a delimited period of endocrine activation during pregnancy can have an adverse effect on infant neurobehavioral development, like that of prenatal stress.


[Effects of suplatast tosilate (IPD-1151T) on antibody formations in mice]. We examined the effects of suplatast tosilate (IPD-1151T) on antibody formation in mice, and the following results were obtained: 1) IPD-1151T clearly increased the productions of IgM and IgG-hemolytic plaque forming cells (PFC) in mice immunized with sheep red blood cells (SRBC) when the agent was given p.o. for 5 days from the day of immunization. 2) IgM and IgG-PFC productions in old mice, 60 weeks of age, were clearly lower than those in adult mice, 10 weeks of age; and an oral administration of IPD-1151T significantly recovered the decayed PFC-production in the old mice. 3) IPD-1151T clearly suppressed the production of anti-dinitrophenyl (DNP)-IgE antibody in mice when the agent was given p.o. for 5 days from the day of immunization, whereas the agent did not affect anti-DNP-IgM and IgG antibody productions. 4) IPD-1151T did not affect the induction phase of the cellular immune reaction of picryl chloride-induced contact dermatitis and the SRBC-induced footpad reaction. Our present results suggest that IPD-1151T has a class-specific suppressive effect on the production of IgE antibody, but does not suppress the other immune responses.


Alterations in femoral bone histomorphometry and vitamin D metabolism in neonatal streptozotocin-induced diabetic rat. Histomorphometric examination and histological observation of femoral bone were performed on long-standing neonatal streptozotocin-induced diabetic rats (n2STZ, n5STZ) as a human model of non-insulin-dependent diabetes mellitus. The growth and strength of femurs decreased in the STZ diabetic rats. Histomorphometric parameters such as cortical bone thickness, number of metaphysical trabeculae and percent trabecular volume of metaphysical area all significantly decreased in the STZ diabetic rats. There were no significant differences in parameters between the n2STZ and n5STZ diabetic rats. Histological findings demonstrated no significant change in the number of osteoclasts in femur nor change corresponding to osteomalacia. Bone absorption in the STZ diabetic rats appeared unchanged. The plasma calcium level did not change in the STZ diabetic rats, although their plasma phosphate or A1-p levels increased. Circulating 24, 25 (OH)2D3 was significantly lower in the STZ diabetic rats than the controls. However, 25 (OH) D3 or biologically active 1, 25 (OH)2D3 was not different between the controls and STZ diabetic rats. Osteopenia is thus present in the femurs of long-standing neonatal STZ diabetic rats, due in part to abnormal vitamin D metabolism.


Effects of ultraviolet-inactivated feline leukemia virus on the production of alpha/beta interferon by feline peripheral blood mononuclear cells. The effects of ultraviolet-inactivated feline leukemia virus (UV-FeLV) on the production of alpha and/or beta interferon (IFN) by mononuclear cells stimulated with the Newcastle disease virus (IFN-NDV) was investigated. The production of IFN-NDV and gamma IFN was suppressed by 50% as compared to the control in the presence of 200 ng/ml or 200 micrograms/ml of UV-FeLV, respectively. The presence of UV-FeLV decreased the rate of the production, but the time to reach the plateau of the IFN activity in the culture supernatant was not shortened in both IFNs. It was suggested that the suppressive effects of UV-FeLV against both IFNs were through rather similar mechanisms. The suppressive effects of UV-FeLV on IFN-NDV production was most evident in the cultures to which UV-FeLV was added at the early stages of the culture, and could not be demonstrated when UV-FeLV was added 8 hr after initiation of the culture. These data therefore indicate that non-infectious feline leukemia viral particles can modify the production of feline IFNs by peripheral blood mononuclear cells.


Anti-hepatitis C virus immunoglobulin M antibody in patients with chronic hepatitis type C. Anti-hepatitis C virus immunoglobulin M antibody titers were measured by enzyme-linked immunosorbent assay in 28 patients with chronic hepatitis C. Nine patients were treated with alpha-interferon at a dosage of 3 MIU/day for 4 weeks. Anti-HCV IgM was positive in 12 of the 28 patients, and these 12 patients also had significantly higher anti-HCV antibody titers than those patients who were negative for anti-HCV IgM. Among the 9 patients treated with alpha-interferon, anti-HCV IgM positivity was significantly lower in 4 good responders than in 2 non-responders and 3 transient responders at 12 months after the completion of treatment (p < 0.01). These results suggest that anti-HCV IgM titers might be useful for monitoring the antiviral effect of interferon treatment in CH C.


Herpes simplex virus types 1 and 2 and cytomegalovirus in peptic ulcer disease and non-ulcer dyspepsia. The prevalence and the serum levels of IgG antibody to Herpes simplex virus type 1 or 2 (HSV1, HSV2) and to cytomegalovirus (CMV) were studied by ELISA in patients with active peptic ulcer -- duodenal and gastric -- and non-ulcer dyspepsia. Two hundred and forty-two consecutive patients with endoscopically confirmed active peptide ulcer -- 170 duodenal ulcers, 72 gastric ulcers -- and 95 consecutive patients who fulfilled the criteria for the diagnosis of non-ulcer dyspepsia were included in the study. The patients, aged 17-80 years, were well matched for age and sex. Antibody to cytomegalovirus was found in 83% of duodenal ulcer, 85% of gastric ulcer and 75% of non-ulcer dyspepsia patients; differences were not significant. The prevalence of HSV1 antibody was significantly higher in patients with duodenal ulcer than in those with non-ulcer dyspepsia (p < 0.025); the prevalence of HSV2 antibody was significantly higher in patients with duodenal or gastric ulcer, than in those with non-ulcer dyspepsia (p < 0.05, p < 0.01, respectively); however, antibody levels (mean optical density) to the viruses studied were similar for all groups of patients. These results provide some evidence that HSV might be implicated in the pathogenesis of peptic ulcer disease.


Hemoglobin Sabine [beta 91 (F7) Leu-->Pro]: occurrence in a Sardinian individual with hemolytic anemia and inclusion bodies. Hemoglobin (Hb) Sabine (beta 91 Leu-->Pro) is an unstable variant detected for the first time in a 16-year-old Scottish-English-German girl affected by moderately severe hemolytic anemia. A second case was described in a patient of Yugoslavian descent. We report another case of this Hb variant arising as a de novo mutation in a Sardinian patient. Definition of the mutation was obtained by DNA direct sequencing on amplified beta-globin gene, as well as by structural analysis of the hemoglobin variant. The patient presented a moderately severe hemolytic anemia with red blood cell inclusion bodies. Hemoglobin electrophoresis showed that quantitatively the abnormal fraction represented 9% of the total Hb amount. beta globin gene analysis revealed a single nucleotide substitution, T-->C, at codon 91, which gives rise to a leucine-->proline substitution. Structural analysis of the variant confirmed the amino acid substitution (Leu-->Pro) predicted by DNA sequencing.


Effect of enalapril treatment and sclerotherapy of esophageal varices on hepatic hemodynamics in portal hypertension. The hemodynamic effects of long-term (3 months) treatment with enalapril, a potent angiotensin converting enzyme inhibitor, were studied in 12 randomly selected patients with portal hypertension and a previous episode of hemorrhage from esophageal varices. All these patients underwent injection sclerotherapy of varices at 1-week intervals. As a control group 13 patients treated only with injection sclerotherapy and placebo were used. After 3 months the wedged hepatic venous pressure (25.5 +/- 4.8 vs. 21.3 +/- 4.8 mmHg) and the non-wedged hepatic venous pressure gradient (17.0 +/- 6.0 vs. 12.6 +/- 3.4 mmHg) were significantly lower than the basal values (p < 0.01) in the group treated with enalapril. A large decrease (> 3 mmHg) in these pressures was observed only in 50% of the patients. In the group treated with sclerotherapy+placebo this pressure reduction was not observed. Systemic hemodynamics and liver function tests did not change during the treatment. None of our patients died during the study or the next 6 months. We conclude that enalapril lowers portal pressure in patients with portal hypertension, although not in all of them, and may be used to good effect to manage patients with esophageal varices in combination with sclerotherapy.


Surgical treatment for hepatocellular carcinoma (HCC) 3 cm or less than 3 cm in diameter. The efficacy of surgical treatment of small hepatocellular carcinoma (3 cm less in diameter) was studied in 30 patients with solitary tumors. The survival of the patients surviving at least three months after resection was 95% at 1 year, 69% at 2 years, 50% at 3 and 4 years. With regard to the range of resection in cases of partial resection the survival rate was 94% after 1 year, 64% after 2 years, and 47% after 3 and 4 years, while in the case of more than subsegmental resection the 4-year survival rate was 100% and a significant difference was noted. Recurrence was noted after 47% of the partial resections and 25% of the subsegmental or larger resections, and 6 of the 8 recurrent site especially in lesions of less than 2 cm were within the same segmental region. Thus, in the surgical treatment of solitary cancers smaller than 3 cm in size, resection of the segmental resection seems to be preferred whenever the functional reserves of the liver permits.


A new look in the management of unresectable primary hepatocellular carcinoma. From January 1992 to October 1992, nine patients with unresectable primary hepatocellular carcinoma were treated either by liver resection combined with transarterial on-target chemotherapy (n = 4) or by transarterial on-target chemotherapy alone (n = 5). All nine patients were seen with diffuse spread of their disease and were considered as refractory to surgical treatment. The patients who had liver resection combined with alcohol transtumoral injection of the residual tumor in the liver remnant and transarterial lipiodol on-target chemotherapy, responded well and were seen postoperatively with a significant decrease in size of their residual tumor, which was found histologically to have advanced necrotic changes. Similarly, the remaining patients, who had only alcohol injection and frequent administration of on-target chemotherapy, were seen with necrosis of their tumor and with decrease in its size. The fetoprotein serum levels were decreased in all patients. None of the patients showed systemic effects from the use of chemotherapy, nor did they demonstrate any hepatotoxic side effects.


[Studies on nuclear DNA content in testicular germ cell tumors using flow cytometry]. Flow cytometric DNA analysis was done in 76 patients of testicular germ cell tumors (GCTs) experienced in our clinic for evaluating the clinical relevance of DNA index (DI), clarifying those biological features and shedding some insights in the pathogenesis of testicular GCTs. Histological type and its incidences were seminomas in 35 adults, nonseminomatous germ cell tumors (NSGCTs) in 24 adults and prepubertal germ cell tumors (P-GCT) in 17 boys (9 in yolk sac tumors and 8 in teratomas). Totally 190 samples of paraffin embedded materials (with a mean of 2.5 samples per case) were histologically reconfirmed and employed the flow cytometry analysis with some modification of the Hedley's technique for analyzing the DNA ploidy and DIs. Coefficient of variation (CV) were acceptably ranged from 3 to 10%. In 58 evaluable cases with adult GCTs, 57 (98%) were revealed as DNA aneuploid, while DNA diploid tumor were observed in only one case with NSGCT. On the other hand, all specimens showed DNA euploid in P-GCT; DNA diploid in all teratomas and 6 yolk sac tumors, DNA tetraploid in other 3 yolk sac tumors. DNA aneuploid pattern was not apparently detected in children. Even in one adult case with pure yolk sac tumor and two adults with mature teratomas, all specimens were classified as DNA aneuploid. These results support the present hypothesis that the pathogenesis of P-GCTs is different from that of adult GCTs. DIs in adult NSGCTs (median DI = 1.56) are significantly lower than those in adult pure seminomas (median DI = 1.85) (p < 0.01). Although there was no significant correlation among the DIs in NSGCTs and the clinical staging of Japanese Urological Association, the distribution of DIs in NSGCT patients of the advanced extent were lower than that of the other extents of NSGCTs on the basis of Indiana University staging system (p < 0.05). In general, it has been postulated that the higher DI is paralleled to the more malignant nature of neoplasms. Nevertheless, this study conversely suggested that the lower DI in adult testicular GCTs is apt to related with high malignant potential determined by histological type and clinical stage. DNA heterogeneity was observed only in 4 of 23 cases with adult NSGCTs (17%), but 3 of these 4 cases (75%) with DNA heterogeneity were assigned as the advanced extent. These data suggest that the lower DI and the presence of DNA heterogeneity may have prognostic relevance for adult NSGCTs.


[Electromechanical responsiveness to alpha 1-adrenoceptor and angiotensin II receptor stimulation in the ventricular myocardium of hypertrophied and dilated cardiomyopathic Syrian hamsters]. It has been postulated that neurohumoral factors, such as sympathetic nervous system or renin-angiotensin system, may play an important role in the development of myocardial lesion in cardiomyopathic hamsters. This study was undertaken to determine whether the electromechanical responsiveness of the ventricular myocardium to alpha 1-adrenoceptor and angiotensin II (Ang II) receptor stimulation is altered in cardiomyopathic (CM) Syrian hamsters. Changes of action potential and isometric tension in response to these receptors stimulation were examined in left ventricular papillary muscles of 14-20 week-old normal (F1 beta), hypertrophic (BIO 14.6) and dilated (BIO 53.58) cardiomyopathic hamsters using conventional microelectrode techniques. Action potential duration (APD) recorded from the papillary muscles (PMs) of BIO 14.6 was significantly shorter than that recorded from BIO 53.58 and F1 beta PMs. Developed tension, which was corrected for the cross-sectional area of PMs, was significantly reduced in PMs of both cardiomyopathic hamsters than those of normal hamsters. Phenylephrine (0.3-30 microM) in the presence of propranolol (1 microM) produced concentration-dependent increases in APD and DT in PMs of normal and cardiomyopathic hamsters. Increases in APD and DT in response to 30 microM phenylephrine (PHE) in BIO 14.6 PMs were significantly greater than those in PMs of F1 beta. However, the increases in APD and DT produced by alpha 1-adrenergic stimulation in BIO 53.58 PMs were comparable to those observed in F1 beta PMs. In order to define ionic mechanism(s) responsible for the greater increase in APD during alpha 1-adrenergic stimulation in PMs of BIO 14.6, effects of PHE on membrane currents were examined in ventricular cells of F1 beta and BIO 14.6 using patch clamp techniques. PHE-induced decrease in the transient outward current (I(to)) in BIO 14.6 cells was similar to that in F1 beta cells. However, PHE produced a significantly greater decrease in the inward rectifier potassium current (IK1) in BIO 14.6 myocytes than in F1 beta ones. The greater inhibition of IK1 may, at least in part, explain the greater prolongation of APD in BIO 14.6 PMs. Angiotensin II (Ang II, 0.01-1 microM) also produced concentration-dependent increases in APD and DT in PMs of normal and cardiomyopathic hamsters. In contrast to electromechanical responses to alpha 1-adrenergic stimulation, Ang II produced a significantly smaller increase in DT in BIO 14.6 PMs than F1 beta ones, concomitantly with a smaller increase in APD. The Ang II-induced increase in DT in BIO 53.58 PMs was also smaller than that in F1 beta PMs.(ABSTRACT TRUNCATED AT 400 WORDS)


Fallacies in the interpretation of Paul-Bunnel Davidsohn differential test. Serological findings in five cases where Paul-Bunnel Davidsohn (PBD) test results were misleading, are presented. Three patients' with Chronic renal disorder and positive PBD test had specific serology results, signifying Cyto Megalo Virus infection. A fourth patient with Hepatitis B Virus infection also had positive PBD test. Forssman type of antibody response was demonstrable in a boy with recent Epstein-Barr virus infection and high Cyto Megalo Virus antibody titres.


Periampullary carcinoma with special reference to their mucin characteristics--a pathologic study of 53 cases. Histological features and mucin characteristics of 53 cases of periampullary carcinoma, a rare group of epithelial tumors is presented. The cases comprised of 17 resected specimens and 36 endoscopic biopsies. Jaundice was the commonest symptom. The mean tumor size was 2 cms diameter (1-6 cms). Forty cases were characterised as well differentiated adenocarcinoma, 5 as moderately differentiated adenocarcinoma, 5 as mucin secreting adenocarcinoma, 3 as papillary adenocarcinoma and 1 as undifferentiated carcinoma. Desmoplasia and insignificant mitosis were amongst the commonly encountered histological features. Forty four cases (83%) stained positive for mucin (PAS-AB) with a majority of them (66%) predominantly showing acidic mucin. The likely prognostic implication of this feature is discussed.


The control of cardiac chronotropic function in hypobaric hypoxia. Altitude hypoxia induces an increase in adrenergic activity in humans. However, a decrease in maximal heart rate is observed after a few days of exposure to altitudes above 3500 m, as well as a decrease in chronotropic response to isoproterenol infusion. This phenomenon has been linked to a desensitization of beta-adrenoceptors (beta AR), and/or an increase in parasympathetic activity. A decrease in the density of beta AR in chronic hypoxia has been found in rat left ventricle and in human lymphocytes, without modification of the affinity of beta AR for an agonist or antagonist, and a decreased adenylate cyclase activity in the right ventricle. In the same conditions, the density of adenosine A1 receptors is decreased by 46% in rat myocardium, without alteration in the coupling between hormone, receptor and Gi protein. The density of muscarinic receptors is increased by 40%, with an increase in the affinity for an agonist, suggesting an augmented parasympathetic effect. Hypoxia probably acts on all the receptors involved in the modulation of cardiac chronotropic activity; the combined effects of chronic hypoxia on these receptors tend to a beneficial limitation of myocardial oxygen consumption, especially during heavy exercise.


Adrenergic system in high altitude residents. Heart rate (HR) response to isoproterenol (ISO) infusion (IP) is decreased in normal sea level (SL) natives exposed to high altitude (HA). Since norepinephrine plasma concentration is higher in HA hypoxia, a downregulation of beta-adrenoceptors (beta AR) was evoked. We explored this phenomenon at 3600 m in a HA normal population (HAN) and in polycythemic subjects (HAP). Results are compared to SL natives in normoxia (SLN), and during chronic hypoxia at 4800 m (SLH) (J Appl Physiol 65:1957-1961, 1988). ISO dose required to raise HR by 25 min-1 (I 25) is not different in HAN or HAP group when compared to SLN. Density of beta AR on lymphocytes was 39% and 25% lower in HAN and HAP than in SLN group, respectively. Chronotropic response to IP is similar in SL and HA subjects under their usual environmental conditions, while SL natives show a blunted response under hypoxia, probably due to a decrease in beta AR density. No adrenergic desensitization was found in highlanders. Lower beta AR density in HA groups could be an adaptive mechanism to chronic hypoxia. Polycythemia does not affect this responsiveness.


Screening for cervical cancer in Latin America: a case-control study. The beneficial effect of cervical cytology in reducing the incidence of invasive cervical cancer is well accepted, but many issues regarding specific patterns of screening remain to be resolved, and preventive programmes need to be adapted to regional characteristics. In a case-control study conducted in Latin America, we investigated cytological screening histories of 759 cases of invasive cervical cancer and 1430 controls, with participation rates of 99% and 96%, respectively. Fifty per cent of the cases and 29% of the controls reported never having been screened. Screening was less common among older, less educated and less parous women; non-users of oral contraceptives and women without histories of venereal diseases. There was also evidence that older women and those with multiple partners had longer intervals between examinations. The relative risk (RR) associated with no prior screening was approximately 3 and was not modified by other risk factors. Women reporting a Pap smear within 24-47 months before interview had the same RR as those examined within 12-23 months. Women tested longer ago had higher risks, but still much lower than women never examined. There was evidence that one examination is associated with less reduction in risk than two, regardless of the interval since last Pap smear. Screening appeared to reduce risk of both squamous cell carcinomas and adenocarcinomas. As expected, cases presenting at advanced stages were less likely to have been screened and reported longer intervals since their last examination. These results support the need to concentrate limited resources in the groups that need screening most, mainly older and less educated women who have never been screened.


A novel, mild, specific and indirect maleimido-based radioiodolabeling method. Radiolabeling of analogs derived from parathyroid hormone (PTH) and PTH-related protein (PTHrP). In an effort to design a mild, non-oxidative and site-specific means of radiolabeling bioactive molecules we have employed maleimido-sulfhydryl chemistry to produce bioactive hormone radioligands. We have prepared two novel radioiodolabeled reagents, 3'-maleimidopropanoyl-3-125I-tyramide and its retro analog, N-maleoyl-N'-3-(4-hydroxy-3-125I-phenyl)propanoyl ethylenediamide, by either oxidative radioiodination of the precursors or radiolabeling of the phenolic component prior to its incorporation into the radiolabeling reagents. These reagents were then used to radiolabel analogs of parathyroid hormone (PTH) and parathyroid hormone-related protein (PTHrP) in an efficient way, yielding reaction mixtures which were easily purified. The radioligands obtained are stable upon storage and bind in a reversible manner to a single population of binding sites displaying affinity in the low nanomolar range. The potencies of these analogs are comparable to the non-modified PTH and PTHrP analogs. This study demonstrates the utility of the novel maleimido-based indirect radioiodination approach and highlights some of its advantages over either direct oxidative procedures or acylation using the Bolton-Hunter reagent.


AZT-induced mitochondrial myopathy.

AZT-induced mitochondrial myopathy. Histochemical, electron microscopy and biochemical studies were performed on muscle biopsy specimens from 11 AIDS patients treated with zidovudine. A peculiar association of structural abnormalities and mitochondrial dysfunction was found. Focal cytochrome c oxidase (COX) deficiency was evident in muscle sections from 9 patients, 8 of whom had received long-term treatment while one had been treated for 1 month only. Electron microscopy showed changes in number, size and structure of mitochondria. Biochemical studies proved partial COX and succinate cytochrome c reductase (SCR) deficiency in 4 patients; one patient had only reduced SCR activity. Our data confirm that AZT therapy can cause toxic myopathy with mitochondrial dysfunction.


Potentiation of topoisomerase I and II inhibitors cell killing by tumor necrosis factor: relationship to DNA strand breakage formation. Recombinant human tumor necrosis factor (rHuTNF) synergistically potentiates the cytotoxicity of the topoisomerase I inhibitor camptothecin, and the topoisomerase II inhibitors epidoxorubicin, etoposide, mitoxantrone, ellipticine, actinomycin D and 4'-(9-acridinylamino)methanesulfon-m-anisidide on A2780 human ovarian cancer cell line. Similar synergy was not observed with a combination of rHuTNF and cis-platinum or mitomycin C. When A2780 cells were incubated with rHuTNF simultaneously with camptothecin or mitoxantrone or VP16, increased numbers of DNA single-strand breaks were produced. rHuTNF alone did not induce DNA strand breakage. These data provide evidence that the enhancing effect of rHuTNF is closely related to the DNA damage mediated by topoisomerase-targeted drugs. These observations may have relevance for ovarian cancer treatment.


Effects of modifying agents on conformity of enzyme phenotype and proliferative potential in focal preneoplastic and neoplastic liver cell lesions in rats. Development of preneoplastic lesions in the rat liver under the influence of various modifiers was investigated with particular attention to changes in simultaneous expression of altered enzyme phenotype within the lesions (conformity) and proliferation potential. Degree of conformity of marker enzymes such as glutathione S-transferase placental form (GST-P), glucose-6-phosphate dehydrogenase (G6PD), glucose-6-phosphatase, adenosine triphosphatase and gamma-glutamyltranspeptidase was compared with levels of 5-bromo-2-deoxyuridine labeling. After initiation with diethylnitrosamine, rats were administered the hepatopromoter sodium phenobarbital (PB, 0.05%), the antioxidant ethoxyquin (EQ, 0.5%), or a peroxisome proliferator, clofibrate (CF, 1.0%) or di(2-ethylhexyl)-phthalate (0.3%) and killed at week 16 or 32. The PB promoting regimen was clearly associated with increase in the numbers of high conformity class lesions simultaneously expressing three to five enzymes, and elevated proliferation potential. The inhibitor, EQ, in contrast, brought about a time-dependent decrease in conformity so that only 1 or 2 alterations were most commonly observed at week 32. Lesion populations in the peroxisome proliferator- and especially CF-treated cases were characterized by obvious dissociation between degree of conformity and proliferative status. Such treatment-dependent differences were not always correlated with the size of the lesion. The results thus suggested that the conformity and proliferation potential of preneoplastic lesions are dependent on modification treatment. Overall, GST-P was found to be the most reliable marker, although G6PD was less influenced in the peroxisome proliferator cases.


Phenotypic properties of liver tumors induced by dehydroepiandrosterone in F-344 rats. Dehydroepiandrosterone (DHEA), a C19 adrenal steroid hormone, induces peroxisome proliferation in liver cells and is hepatocarcinogenic in the rat. The present study deals with the phenotypic properties of DHEA-induced liver lesions. A majority of the altered areas (80-87%), neoplastic nodules (> 94%) and hepatocellular carcinomas (HCC, 80-100%) lacked the marker enzymes gamma-glutamyltranspeptidase and placental form of glutathione S-transferase (GSTP). Northern blot analysis of HCC from 4 rats revealed no detectable GSTP mRNA. These HCC, however, showed a marked decrease in the staining of glucose-6-phosphatase and adenosine triphosphatase. These results indicate that the phenotypic properties of liver tumors induced by DHEA and amphipathic carboxylate peroxisome proliferators are similar.


Frequent occurrence of p53 gene mutations in uterine cancers at advanced clinical stage and with aggressive histological phenotypes. The clinical and pathological significance of mutation of the p53 tumor-suppressor gene was examined in 108 cases of primary uterine cancers using single-strand conformation polymorphism and direct DNA sequencing analyses. Mutation of the p53 gene was detected in 19 (31%) of 62 cases of cancer of the uterine corpus and was more frequent in groups at an advanced clinical stage and/or with aggressive histology. Among four adenocarcinomas arising in the lowest portion of the uterine corpus, three showed integration of human papillomavirus (HPV) types 16 and/or 18 DNA, and two of them also showed p53 mutation. In cancer of the uterine cervix, p53 mutations were rare; 7% (3/46) in total, 3% (1/30) of cases with integration of HPV types 16 and/or 18 DNA and 13% (2/16) of cases without HPV DNA integration. Three mutations were detected among two cases at clinical stage IV and two cases of undifferentiated cervical carcinoma. Immunohistochemically, all five cases of uterine cancer which showed diffuse (> 50% of cancer cells) nuclear staining of p53 protein also carried the p53 mutation. Therefore, p53 alterations were suggested to be involved in the development of uterine cancers showing aggressive biological behavior. Although a high incidence of HPV DNA integration and a low incidence of p53 mutation were confirmed in cancer of the uterine cervix, there was no inverse association between integration of HPV types 16 and/or 18 DNA and p53 mutation.


Evidence of direct generation of oxygen free radicals from heterocyclic amines by NADPH/cytochrome P-450 reductase in vitro. Highly purified cytochrome P-450 reductase (also called cytochrome c reductase; EC 1.6.2.4.) and NADPH were used to generate superoxide radical (O2.-) from 11 different heterocyclic amines (HCAs) as identified by electron spin resonance spectroscopy using the spin trapping method with 5,5-dimethyl-1-pyrroline N-oxide (DMPO). The signal intensity of DMPO-OOH(-O2-) (i.e., the DMPO spin adduct of O2.-) was strongest for 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ). The O2.- generation with HCAs decreased in the following order: 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx) = 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) > 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (diMeIQx) > or = other HCAs; O2.- generation was lowest with 2-amino-3-methyl-9H-pyrido[2,3-b]indole .CH3COOH (MeA alpha C). By using Lineweaver-Burk plots, Km values of cytochrome P-450 reductase for mitomycin C, IQ, and MeIQ were determined to be 1.60 x 10(-6) M, 1.97 x 10(-5) M, and 2.83 x 10(-6) M, respectively. The present findings have important implications for carcinogenesis because of the known effect of oxygen radicals on cell proliferation.


Differential changes in expression of gap junction proteins connexin 26 and 32 during hepatocarcinogenesis in rats. We examined expressions of the gap junction proteins, connexin 26 (Cx26) and 32 (Cx32), in preneoplastic and neoplastic lesions during rat hepatocarcinogenesis. A marked reduction in the number of Cx32-positive gap junctions was observed in 17% of the glutathione S-transferase placental form-positive foci, whereas 44% of the foci showed increased expression of Cx26. Most hyperplastic nodules exhibited decreased expression of Cx32, whereas 16% of the nodules showed increased expression of Cx26. In hepatocellular carcinomas, expressions of both Cx32 and Cx26 were significantly reduced. These results suggest that the expressions of Cx32 and 26 are differentially regulated during hepatocarcinogenesis, and that the decrease in Cx32 expression occurs earlier, whereas reduction in Cx26 expression occurs later in association with promotion and progression of carcinogenesis.


Retrovirus-mediated gene transfer targeted to malignant glioma cells in murine brain. A murine model for meningeal metastasis of malignant glioma was developed to study selective gene transfer into tumor cells and to establish a reliable means of determining the rate of tumor cell infection. A murine ecotropic retroviral vector was created in which the Escherichia coli beta-galactosidase gene served as a marker for gene expression from the integrated retrovirus. This retrovirus exhibited a high rate of infectivity in RSV-M mouse glioma cells in vitro. The recombinant retrovirus was injected directly into the cisterna magna of the mice. Staining of beta-galactosidase showed that the rate of gene integration was high in the disseminated glioma cells. These results suggest the possibility of retrovirus-mediated gene therapy for meningeal dissemination of malignant glioma.


Human lung cancer cell line producing hepatocyte growth factor/scatter factor. Hepatocyte growth factor (HGF)/scatter factor (SF) is a cytokine which is produced by mesenchymal cells and stimulates the motility of some epithelial cells, including cancer cells and vascular endothelial cells. Two human lung cancer cell lines, PC-1 and PC-13, were found to produce a protein which was indistinguishable from HGF/SF with regard to biological activities and immunological characteristics, although they were derived from epithelial cells. In general, highly aggressive cancer cells often show some mesenchymal characteristics, and production of HGF/SF by cancer cells is also considered as a phenomenon of acquisition of mesenchymal phenotype, which may be involved in cancer invasion and progression. These cell lines showed no apparent response to exogenous HGF/SF. In addition, no c-met proto-oncogene product was detectable in these cells by Western blot analysis. Although the function of HGF/SF produced by cancer cells, either autocrine or paracrine stimulation, remains to be studied, this is the first report to describe cancer cells producing HGF/SF.


Pharmacokinetic study of mitomycin C with emphasis on the influence of aging. Mitomycin C (MMC) at a dose of 8 mg/m2 was administered by short intravenous infusion to 14 cancer patients. All patients had normal renal, hepatic, cardiac and bone marrow functions. The plasma level of MMC, which was determined by high-performance liquid chromatography over a 24-h period after the infusion, fitted the 2-compartment model curve except for one patient. There was a significant correlation between the area under the time versus concentration curve (AUC) of MMC and the age of patients (r = 0.558, P < 0.05). Pharmacokinetic parameters in one of the older (a 69-year-old male) patients were extremely different from those of the other 13 patients. This patient experienced severe myelosuppression, probably due to the markedly increased AUC of MMC. Our results suggest that a patient's age has a significant influence on the pharmacokinetics of MMC in patients with normal major organ functions and that in some patients, MMC pharmacokinetics may be altered, possibly due to interpatient variation in the activation or metabolic pathway of MMC.


Aplastic crisis and leg ulceration: two rare complications of hereditary sideroblastic anaemia. Aplastic crisis as a result of parvovirus infection is seen in a number of haematologic disorders characterized by decreased red cell survival, and leg ulceration due to unknown causes is seen in a number of haemolytic anaemias. Neither of the two has been reported in a case of sideroblastic anemia. We report one case with each of these complications in association with sideroblastic anaemia.


High-performance liquid chromatographic assay for the alpha-melanotropin[4,10] fragment analogue (Melanotan-II) in rat plasma. A high-performance liquid chromatographic (HPLC) procedure has been developed for the quantification of Melanotan-II (MT-II), a cyclic heptapeptide which promotes rapid tanning of the skin, in rat plasma. The method involves precipitation of plasma proteins followed by direct-injection HPLC with ultraviolet detection. Calibration curves were linear over the range 100-1000 ng/ml for rat plasma. The method is reproducible and reliable with a detection limit of 50 ng/ml in plasma. Within- and between-day precision and accuracy reported as coefficient of variation and relative error, respectively, were < 7%. The application of the assay was successfully demonstrated by quantifying the concentration of MT-II in rat plasma samples following an intravenous dose of 0.3 mg/kg.


Determination of stavudine, a new antiretroviral agent, in human plasma by reversed-phase high-performance liquid chromatography with ultraviolet detection. A sensitive high-performance liquid chromatographic assay has been developed to determine the levels of a new antiretroviral agent, stavudine (2',3'-didehydro-3'-deoxythymidine, d4T), in human plasma. Didanosine (2',3'-dideoxyinosine, ddI) was used as the internal standard. The very selective sample pretreatment involved solid-phase extraction using silica gel columns. Chromatography was carried out on a mu Bondapak phenyl column, using a mobile phase of 0.005 M phosphate buffer (pH 6.8)-methanol (90:10, v/v) and ultraviolet detection at 265 nm. The method has been validated, and stability tests under various conditions have been performed. The detection limit is 10 ng/ml (using 500-microliters human plasma samples). The bioanalytical assay has been used in a single pharmacokinetic experiment in a rat to investigate the applicability of the method in vivo.


Transneuronal transfer of herpes simplex virus type 1 (HSV 1) from mixed limb nerves to the CNS. I. Sequence of transfer from sensory, motor, and sympathetic nerve fibres to the spinal cord. The time course of transneuronal transfer of Herpes simplex virus type 1 (HSV 1) from sensory, motor, and sympathetic nerve fibres to connected spinal neurones was examined. After injection of a constant number of infectious units into distal forelimb or hindlimb nerves of inbred rats of the same age, the extent of viral transfer was strictly dependent on the survival time postinoculation (p.i.). Retrograde transport to somatic motoneurones occurred at 28-29 hours p.i. (stage 1), in synchrony with anterograde transneuronal transfer via small cutaneous afferents (to laminae I-II). At 36-43 hours p.i. (stage 2), retrograde transneuronal transfer from sympathetic nerve fibres first labelled sympathetic preganglionic neurones. At 48-51 hours p.i. (stage 3), transfer via sensory and sympathetic axons became more extensive, labelling laminae III-IV and other preganglionic neurones. Transneuronal transfer from large muscle afferents and motoneurones (to Clarke's columns and the spinal intermediate zone) occurred only at 66-78 hours p.i. (stage 4). Further increases in distribution (stages 5-6) obtained between 78 and 97 hours p.i. may reflect both specific labelling of second and third order neurones and a gradual local loss of specificity. These results indicate that transfer of HSV 1 occurs through all main classes of peripheral axons, but that both anterograde and retrograde transneuronal transfer from small (unmyelinated and fine myelinated) cutaneous and sympathetic axons precedes transfer from large (myelinated) cutaneous and muscle afferents and motor axons. Analysis of viral transfer at sequential intervals is required to distinguish serially connected neurones, determine the route of labelling, and ensure its specificity.


Squamous cell carcinoma of the gingiva found in a patient with AIDS. A young woman with AIDS developed squamous cell carcinoma of the gingiva. This malignancy rarely develops in young people and may indicate an underlying condition.


Detection of hepatitis C virus RNA by nested polymerase chain reaction in sera of patients with chronic non-A, non-B hepatitis treated with interferon. To evaluate the effect of interferon-alpha treatment on the levels of serum aminotransferase (sALT) and of hepatitis C virus (HCV)-RNA, we studied 19 patients with chronic non-A, non-B (NANB) hepatitis. Before therapy, 14 patients were positive by nested polymerase chain reaction (PCR) with primers deduced from the 5'-non-coding region of the HCV genome. Serum HCV-RNA had disappeared in 12 (85.7%) of them by the end of therapy, but then reappeared 6 months later in 4 of these 12 patients. A marked improvement in sALT was seen in 5 of the 8 patients with sustained HCV-RNA disappearance, but not in the 4 patients with only transient HCV-RNA negativity. Pre-treatment levels of hepatitis C viremia, analyzed by single PCR and dot blot hybridization, ranged from 2 x 10(3) to 2 x 10(8) copies/ml, and were below 2 x 10(5) copies/ml in patients with a complete response to interferon therapy. These results suggest that this HCV-RNA assay, combined with sALT testing, may be useful for estimation of the long-term efficacy of interferon therapy in hepatitis C.


Hepatic macrophage malfunction in rats with obstructive jaundice and its biological significance. The present study was designed to investigate the pathophysiology of obstructive jaundice by analyzing the function of hepatic macrophages and their role in immune responses and homeostasis in rats. The phagocytic index, determined by the rate of disappearance of 51Cr-endotoxin from the peripheral blood after intravenous injection, was increased in obstructive jaundice 2 weeks after bile duct ligation. The superoxide production of isolated hepatic macrophages and peripheral blood monocytes, measured by the superoxide dismutase inhibitable ferricytochrome c reduction method, was increased. Prostaglandin E2 release, measured by RIA, was markedly increased in rats with obstructive jaundice, but there was no significant difference in interleukin-1 release between jaundiced and control rats. The flow-cytometric analysis of surface molecules of hepatic macrophages showed decreased expression of interleukin-2 receptor in rats with obstructive jaundice. Thus, the functions of hepatic macrophages in rats with obstructive jaundice were impaired. This malfunction may disturb the immunoregulatory network and metabolism, although the exact implications of the altered function of hepatic macrophages have not yet been clarified.


Peripheral and cardiovascular autonomic impairment in chronic liver disease: prevalence and relation to hepatic function. We have evaluated autonomic function using standard cardiovascular tests and a test of peripheral autonomic denervation, the acetylcholine sweatspot test, in 104 patients with biopsy proven chronic liver disease and 35 age- and sex-matched controls. Cardiovascular autonomic dysfunction was significantly more frequent in advanced liver disease compared with early liver disease (71.8% Child B or C vs. 39.7% Child A; p < 0.0006), and a strong correlation between the number of abnormal tests and Child-Pugh score could be demonstrated (Rs = 0.5; p < 0.0001). On multiple logistic regression analysis, cardiovascular autonomic dysfunction was related to age and to Child-Pugh score and occurred independently of the aetiology of liver disease. Peripheral autonomic denervation was found in 39% of patients, was significantly associated with cardiovascular abnormalities (p < 0.009) and correlated with the number of abnormal cardiovascular tests in each patient (Rs = 0.48; p < 0.0001). In chronic liver disease, the prevalence and severity of cardiovascular autonomic dysfunction is related to the severity of hepatic dysfunction and is independent of aetiology, suggesting a common pathogenetic basis related to hepatic damage; the association with peripheral autonomic denervation indicates that at least some of the abnormalities may be due to a true autonomic neuropathy. The possible significance of these findings to the complications of cirrhosis is discussed.


Serum hepatitis C virus RNA and hepatitis B virus DNA in non-A, non-B post-transfusional and sporadic chronic hepatitis. The sera of 36 French patients with post-transfusional and sporadic non-A, non-B chronic hepatitis were investigated, for HBV and HCV infections using a combination of serological and polymerase chain reaction (PCR) assays. Anti-HCV was detected in 75% (27/36) of the patients by ELISA1 and/or RIBA2 tests. HCV-RNA sequences were found in 75% (27/36) of the sera by a single step PCR, using a set of primers located in the 5' non-coding region. Altogether, 89% (32/36) of the patients were found positive with serological and/or molecular tests. Among the positive patients, 68% (22/32) were found positive for both anti-HCV and HCV-RNA, 16% (5/32) and 16% (5/32) were found positive for either anti-HCV or HCV-RNA, respectively. HBV-DNA sequences were detected in two patients associated to the HCV viraemia. This study confirms the extremely high prevalence of HCV infection in NANB chronic hepatitis in France. It also shows possible co-infection by HCV and HBV in hepatitis.


DNA ploidy pattern in synchronous and metachronous hepatocellular carcinomas. DNA ploidy of hepatocellular carcinoma (HCC) was studied in 28 patients using a flow cytometric method. Fourteen patients had two HCCs synchronously, and the remaining 14 had tumor recurrence in the remnant liver 3-41 months after curative resection of primary HCCs. DNA ploidy pattern and histopathologic parameters were compared between the synchronous and metachronous HCCs. Among those with synchronous HCCs, both tumors were diploid in 7 cases and aneuploid in 2 instances. Five patients had HCCs of different DNA ploidy pattern. On the other hand, 5 of 14 patients with metachronous HCCs had a consistent DNA ploidy between primary and recurrent tumors. In 4 cases, the first tumor was diploid whereas the recurrent HCC was aneuploid or tetraploid. In the remaining 5 cases, the primary HCC was aneuploid, but the recurrent tumor was diploid. Assuming that the difference in DNA ploidy pattern indicates a different clonal origin, the current results indicate that at least 36% of synchronous HCCs and 64% of recurrent HCCs develop in a multicentric fashion.


Detection of hepatitis C virus (HCV) RNA sequences in liver tissue by in situ hybridization. In situ hybridization was used to identify the cell types infected by hepatitis C virus (HCV) in the liver. Using an antisense HCV-RNA probe from the 5' non-coding region, HCV-RNAs molecules were detected in liver sections of 4/11 patients with chronic hepatitis C. These 4 positive subjects were also infected by HIV. HCV-RNA-positive strands were detected in scattered hepatocytes as well as in cells identified as mononuclear cells within the inflammatory infiltrates. HCV-RNA negative strands, likely replicative intermediates, were also detected in these cells. This study therefore indicates that replication of HCV may occur in both hepatocytes and mononuclear liver cells.


Encephalopathy and neuropathy in end-stage liver disease before and after liver transplantation. The nervous system involvement of 8 patients with end-stage liver disease was evaluated by means of clinical neurological, neuropsychological, neurophysiological and neuroradiological investigation before and 6-12 months after a successful liver transplantation. Preoperatively, all subjects (7 women, 1 man; mean age 40 years, range 30-54 years) exhibited decreased muscle strength and 2 patients manifested clinical signs of polyneuropathy. In neuropsychological tests, slight visuoconstructive apraxia, and disturbances of verbal memory and cognitive function were observed. Magnetic resonance imaging (MRI) revealed cerebral lesions in two patients. After transplantation, muscle strength reverted to normal in all patients, polyneuropathy improved and in all but 2 patients recovery of neuropsychological functioning was observed. Clinical signs of encephalopathy had disappeared. All patients were emotionally better adjusted after transplantation. Four subjects showed new, albeit mild changes in neurophysiological and neuropsychological tests postoperatively. We conclude that the majority of neurological impairment disappeared after liver transplantation. We want to stress that evaluation of neurological sequelae of liver transplantation needs to be based on assessments both before and after liver transplantation.


Hepatocellular carcinoma. Prognostic factors and survival analysis in 135 Italian patients. This is a retrospective study to evaluate the history of hepatocellular carcinoma and find the relationship between clinical, biochemical and ultrasonographic features and survival in Italian patients. In 135 consecutive patients median follow-up was 16 months (range 1-66 months) and median survival from the time of diagnosis was 12 months. Univariate analysis showed that individual variables associated with significantly decreased survival included: absence of therapy, Okuda's Stage III, Child-Pugh's Class C, alpha-fetoprotein greater than 400 ng/ml, presence of symptoms, moderate or severe ascites, tumor involving both lobes, mixed internal echo pattern, and multinodular or massive type. Multiple regression analysis (Cox model) revealed that the mixed internal echo pattern of hepatocellular carcinoma, the presence of moderate or severe ascites and Okuda's Stage III were independent predictors of high risk of death. These data can help in selecting patients whose probability of survival is considered high enough to undergo treatment and may be useful for stratifying patients in randomized controlled trials.


A plea for consistent reliability in ambulatory blood pressure monitors: a reminder. To compare three different software versions [read-only memory (ROM) versions 1.22, 1.24 and 1.28] of the SpaceLabs model 90202 ambulatory blood pressure monitor. Simultaneous measurements in a two-arm crossover design. Ten measurements each in 14 normotensive persons were performed to compare each pair of monitors. The results were corrected according to the deviation from a mercury column. The systolic (SBP), diastolic (DBP) and mean arterial blood pressure (MAP) was noted and the factor K = (MAP - DBP)/(SBP - DBP) was calculated. The K factor in the two older ROM versions was close to 0.25, with no statistically significant difference. In contrast the K factor was higher in the most recent ROM version than in one of the previous versions. This could be explained by a 5.4 mmHg increase in MAP found by the new version. A significant difference in blood pressure was observed even between two monitors with the same ROM version. Unnotified changes in the software version in the SpaceLabs 90202 monitor operating by an oscillometric technique seriously affect the reliability of MAP measurements. Whenever possible in clinical trials the same monitor should be used for the same patient on each occasion. The industry should inform clinicians of the consequences of updating apparently identical monitors.


Vasopressin gene transcripts in mineralocorticoid hypertension: an in situ study. To test the hypothesis that enhanced expression of the vasopressin gene accompanies the development of deoxycorticosterone acetate (DOCA)-salt hypertension in the rat and to compare the response with those observed during chronic hypernatremia. Transcript levels were determined by measurement of vasopressin messenger RNA (mRNA) in the supraoptic nucleus and paraventricular nucleus by in situ hybridization, autoradiography and image analysis. Plasma, urinary and pituitary vasopressin were determined by radioimmunoassay. High-resolution localization and measurement of specific mRNA in the supraoptic and paraventricular nuclei before and during development of DOCA-salt hypertension were compared with corresponding results in both age-matched controls and normal rats that drank hypertonic saline. Vasopressin mRNA levels were increased in the paraventricular nucleus during the established and chronic stages of DOCA-salt hypertension, but were unchanged in the supraoptic nucleus. Urinary excretion of vasopressin was increased in the prehypertensive, established and chronic phases of DOCA-salt hypertension, whereas plasma vasopressin levels were increased only in the chronic phase. Pituitary vasopressin levels were unchanged. In comparative studies, vasopressin mRNA levels in both the supraoptic and paraventricular nuclei and plasma vasopressin were significantly increased in normal rats drinking 2% saline. Whereas hypernatremic rats showed markedly elevated vasopressin transcripts in the supraoptic and paraventricular nuclei, DOCA-salt hypertension is associated with increased vasopressin mRNA in the paraventricular but not the supraoptic nucleus. The response in the paraventricular nucleus may explain part of the increased peripheral vasopressin levels and suggests that this nucleus makes a critical contribution to the pathogenesis of DOCA-salt hypertension.


Contribution of catecholaminergic neurons of the dorsomedial and ventrolateral medulla oblongata to the hypotensive effect of clonidine in spontaneously hypertensive rats: in vivo voltammetric studies. Our previous electrochemical studies in the normotensive Wistar-Kyoto (WKY) rat showed a positive correlation between the hypotensive effect of low doses of clonidine (2-10 micrograms/kg intravenously) and inhibition of the activity of catecholaminergic neurons within the brainstem, and that this action was mediated by imidazoline-preferring receptors. In the present study the possibility of a relationship between the centrally mediated hypotensive effect of clonidine and the metabolic activity of catecholaminergic neurons of the ventrolateral and dorsomedial medulla oblongata was investigated in pentobarbital-anaesthetized spontaneously hypertensive rats (SHR). Neuronal metabolic activity was monitored by in vivo electrochemistry in the nucleus reticularis lateralis region of the ventrolateral medulla and in the nucleus tractus solitarii region of the dorsomedial medulla oblongata. Hypotensive doses of intravenously administered clonidine (10 micrograms/kg) failed to inhibit the neuronal metabolic activity of the nucleus reticularis lateralis region; fivefold higher doses were required to inhibit these neurons. The low dose of clonidine (10 micrograms/kg) decreased the metabolic activity of the nucleus tractus solitarii region. Nevertheless, this effect did not appear to be correlated with a reduction in blood pressure and it was not attenuated by a prior central injection of idazoxan (5 nmol/kg intracisternally), which almost completely prevented the hypotensive and the neuronal inhibitory effects of clonidine in the nucleus reticularis lateralis region of the normotensive rat. Both the site and the mechanism of the central hypotensive action of clonidine in the SHR appear to be different from those in normotensive control WKY rats.


Effects of dietary salt on sodium-calcium exchange and ATP-driven calcium pump in arterial smooth muscle of Dahl rats. To compare the effects of dietary salt on sodium-calcium exchange and the ATP-driven calcium pump in arterial smooth muscle between Dahl salt-sensitive (DS) and salt-resistant (DR) rats. Aortic rings freshly excised from 16 DS rats and 16 DR rats on a low- (0.3%) or high- (8%) NaCl diet for 4 weeks were superfused with physiological saline and isometric tension was measured. In the presence of 10 mumol/l phentolamine, 10 mumol/l verapamil and 5 mmol/l caffeine, relaxation of a low-Na+ contraction was promoted by external calcium removal. On the high-salt diet, the rate of relaxation at 1.2 mmol/l extracellular sodium (calcium extrusion by calcium ATPase) was significantly lower in aortic rings from DS rats than from DR rats. The increase in the rates of relaxation from 1.2 mmol/l to normal (139.2 mmol/l) extracellular sodium (calcium extrusion by sodium-calcium exchange) was significantly greater with the high-salt diet than with the low-salt diet in rings from DR rats, but it was not different between the high- and low-salt diets in DS rats. The rate of increase in tonic tension by reducing extracellular Na+ from normal to 1.2 mmol/l in the presence of verapamil, caffeine and phentolamine (calcium entry by sodium-calcium exchange) was significantly lower in rings from DS rats than in those from DR rats on the high-salt diet. These observations suggest that the effects of dietary salt on the sodium-calcium exchange system in arterial smooth muscle differ between DS and DR rats and that calcium extrusion by the calcium pump is decreased in DS rats compared with DR rats. The lack of an increase in sodium-calcium exchange in salt-fed DS rats might lead to an elevation in cellular calcium and contribute to the mechanism of hypertension.


Influence of the renal medulla and early treatment with enalapril upon the development of hypertension in young spontaneously hypertensive rats. To investigate the role of the renal medulla in early hypertension in spontaneously hypertensive rats (SHR), and to explore whether the attenuated increase of pressure induced by enalapril treatment is affected by chemical medullectomy. Forty-four male SHR were studied from 5 to 18 weeks of age: 22 remained intact; 22 were medullectomized at 5.5 weeks of age with 2-bromoethylamine hydrobromide; 11 of each of these two groups were treated with enalapril from 6 to 12 weeks of age. Blood pressure, heart rate and body weight were recorded intermittently, and at 18 weeks renal function was also analysed. The results indicate a protective effect of the renal medulla against severe pressure rises in SHR, although even when enalapril also lowered blood pressure in medullectomized SHR, persistent improvements of glomerular filtration rate and renal flow conductance occurred only in intact SHR. Furthermore, after enalapril treatment ended blood pressure rose to higher levels in medullectomized SHR, despite greater sodium-water losses. The renal medulla seems to exert a protective role both during and after enalapril treatment.


Enhanced angiotensin converting enzyme binding in arteries from spontaneously hypertensive rats. To localize and measure angiotensin converting enzyme (ACE) in different vascular beds of genetically hypertensive rats. Quantitative autoradiography using the angiotensin converting enzyme (E.C. 3.4.15.1) inhibitor [125I]351A. [125I]351A binding was significantly increased in the ascending aorta (both adventitia and intima), descending (abdominal) aorta, carotid artery and coronary arteries of adult, 12-week-old spontaneously hypertensive rats (SHR) compared with Wistar-Kyoto (WKY) rats. Increased [125I]351A binding was also present in the descending aorta of 1-week-old SHR compared with age-matched WKY rats, and both groups of young rats had much higher binding than adult rats. No difference in [125I]351A binding was found in the caudal (tail) artery of adult SHR compared with WKY rats. In both the atria and the ventricles of adult SHR, [125I]351A binding was very significantly reduced. Our results indicate that higher ACE concentrations occur in some arteries of genetically hypertensive rats, and support the hypothesis that local arterial concentrations of ACE affect the development and maintenance of genetic hypertension.


Changes in cardiovascular mass, left ventricular pumping ability and aortic distensibility after calcium antagonists in Wistar-Kyoto and spontaneously hypertensive rats. To determine the effects of different dihydropyridine calcium antagonists on cardiovascular mass and function in normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). The rats were treated daily for 3 weeks with nitrendipine (20 mg/kg), nifedipine (30 mg/kg), nisoldipine (6 mg/kg) or their vehicles. At the conclusion of that period left ventricular pumping ability and aortic distensibility were determined, and the aortic, cardiac and left and right ventricular masses. Each drug reduced arterial pressure in both rat strains; each decreased left ventricular mass in SHR but not in WKY rats. All three agents increased right ventricular mass in WKY rats; only nisoldipine did so in SHR. Each compound improved left ventricular pumping ability in WKY rats, maintaining function even when pressure was abruptly increased to pretreatment levels. In contrast, although all three calcium antagonists improved cardiac performance in SHR at the pharmacologically reduced pressures, pumping ability was not maintained when pressure was increased to pretreatment levels in nisoldipine-treated SHR. All three agents improved aortic distensibility in both strains, but only in SHR was reduced aortic mass demonstrated. These data not only continue to demonstrate a functional/structural dissociation associated with antihypertensive therapy, but also suggest subtle functional and structural effects that differ even within the same class of calcium antagonists.


Anatomical localization and pharmacological activity of mature endothelins and their precursors in human vascular tissue. We developed four selective rabbit antisera in order to compare the distribution of immunoreactive mature endothelins and their precursors, proendothelin-1, proendothelin-2 and proendothelin-3, in the endothelium from human vascular tissue. Our second aim was to use in vitro pharmacological assays to test the vasoconstrictor actions of the mature endothelin and proendothelin peptides. The antisera were shown to be selective by enzyme-linked immunosorbent assays. With these antisera, we detected immunoreactivity in serial cryostat sections from saphenous and mesenteric veins, and mesenteric and internal mammary arteries, using a peroxidase-antiperoxidase technique. In pharmacological experiments, segments of human coronary and mesenteric arteries were exposed to cumulative (0.06-60 nmol/l) concentrations of the endothelins and their precursors. Antisera directed against mature endothelin stained the cytoplasm of endothelial cells in all vessels tested. Immunoreactive proendothelin-1 and proendothelin-2 were also detected, but not proendothelin-3. Endothelin-1 and endothelin-2 were strongly vasoactive, with similar molar potencies, and caused a dose-related increase in contractile force in human coronary arteries (0.06-60 nmol/l). However, proendothelin-1 and proendothelin-2 were 100-fold and 1000-fold less vasoactive than their respective mature peptides. No contractile effect was seen with proendothelin-3 or endothelin-3 at the concentrations tested in human coronary arteries, and similar results were obtained with human mesenteric arteries. These results suggest that proendothelin-1 and proendothelin-2 must be converted to their corresponding mature peptides to produce vasoconstrictor activity in human vessels. Immunoreactive mature endothelin is widely distributed in human vascular endothelial cells and, if released, may produce endothelin-mediated vasoconstriction.


Prazosin blunts the antinatriuretic effect of circulating angiotensin II in man. The present study examines the role of alpha 1-adrenoceptors in determining the renal haemodynamic and sodium excretory responses to a physiological dose of angiotensin II in man. The effects of a low-dose infusion of angiotensin II (1 ng/kg per min) and a non-depressor dose of prazosin (0.25 mg), alone and in combination, on urinary sodium excretion (UNaV), effective renal plasma flow (ERPF), glomerular filtration rate (GFR) and segmental tubular function were studied in eight normal male subjects. Subjects were studied undergoing maximal water diuresis. Clearances of inulin and para-aminohippurate were employed to estimate GFR and ERPF, respectively. Segmental tubular handling was assessed by both lithium clearance (CLi) and solute-free water methods. Angiotensin II decreased UNaV without altering ERPF and GFR. Angiotensin II caused a significant fall in fractional CLi, which may indicate a proximal tubular effect of angiotensin II. Angiotensin II alone also increased fractional reabsorption of sodium delivered to the distal nephron, as evaluated by both the CLi method and by estimation of solute-free water clearance. When angiotensin II was given in combination with prazosin, which on its own had no apparent effects on any renal parameters, the antinatriuretic and tubular effects of angiotensin II were significantly blunted. These findings suggest that low doses of circulating angiotensin II are able to modulate UNaV by increasing sodium reabsorption in the proximal and, to some extent, the distal nephron segment in man. The study also showed that a non-depressor dose of prazosin blunted the renal effects of angiotensin II, thereby providing tentative evidence of a renal interaction between alpha-adrenoceptors and angiotensin II in man.


Effect of dietary sodium on platelet alpha 2-adrenoceptors in young normotensive men with or without a family history of hypertension. To study the effects of genetics on the response of platelet alpha 2-adrenoceptors to a change in salt intake. Biochemical measurements and radioligand binding assays in platelets were performed in 11 normotensive male university students with a family history of essential hypertension (FH+) and in 17 students without a family history of hypertension (FH-). The 28 students were fed a high-sodium diet for 7 days and a low-sodium diet for 7 days. In FH+ subjects the number of alpha 2-adrenergic receptors on platelet membrane fractions increased significantly from the high-sodium diet to the low-sodium diet, even though plasma noradrenaline concentrations tended to increase with the low-sodium diet. There was no change in the number of alpha 2-adrenoceptors in the FH--group. In both groups the radioligand binding affinity was decreased during a low-sodium period compared with in a high-sodium period. In the FH+ subjects the change in platelet alpha 2-adrenoceptors associated with altered sodium status was similar to that seen in patients with salt-sensitive hypertension, suggesting that there is a genetic susceptibility to sodium.


Potassium supplementation reduces clinic and ambulatory blood pressure in elderly hypertensive patients. To determine the effects of potassium chloride 60 mmol/day supplementation on clinic and 24-h ambulatory blood pressure values in elderly untreated hypertensive patients. A double-blind randomized placebo-controlled crossover study lasting 8 weeks, following a 4-week run-in period. Outpatient clinic in a district general hospital. Eighteen untreated elderly hypertensive patients (mean age 75 years, range 66-79) with a systolic blood pressure of > or = 160 mmHg and/or a diastolic blood pressure of > or = 95 mmHg were recruited from the clinics of local general practitioners and from the current hospital outpatient department. Patients had not received any antihypertensive medication for at least 4 weeks before entry into the study. Before entry into the study, the daily dietary electrolyte intake of each individual was established and this was maintained during the run-in and intervention periods. Following a 4-week run-in period patients received potassium supplements or matching placebo, each for 4 weeks. The within-patient changes in clinic and 24-h ambulatory blood pressures at the end of each intervention period. After 4 weeks potassium supplementation compared with placebo there was a significant fall in supine clinic blood pressure, standing and 24-h ambulatory systolic blood pressure. There was no significant change in clinic standing diastolic blood pressure, 24-h ambulatory diastolic blood pressure or pulse rate. Plasma renin activity increased and body weight fell after potassium supplementation. Twenty-four-hour urinary potassium rose significantly, whereas urinary sodium excretion was unchanged. A 60-mmol daily supplement of potassium chloride reduces clinic and 24-h ambulatory blood pressure in elderly hypertensive patients.


Quality of life in hypertensives treated with atenolol or captopril: a double-blind crossover trial. To compare the effects of captopril and atenolol on quality of life of hypertensive patients. In a randomly allocated double-blind crossover trial with two 6-week treatment periods captopril at 25 mg twice a day or atenolol at 50 mg once a day were administered to 265 hypertensive patients (mean age 56 years; 55% men). Of these, 65% were newly treated hypertensives and 35% were previously uncontrolled on a diuretic alone. A seated diastolic blood pressure of 95-115 mmHg was required after a 3-week placebo run-in period. Any previous diuretic therapy was changed to hydrochlorothiazide (25 mg once a day) and the dose was kept constant throughout the trial. Newly diagnosed patients did not take a diuretic at any time. Quality of life was assessed from self-completed questionnaires measuring psychological well-being, symptomatic side effects of treatment, and activity and perceived well-being (a health index). A relative's perception of the patients' mood was also obtained where possible. Twelve patients withdrew on atenolol and 10 on captopril. No differences between the drugs were observed in quality of life measures, and 95% confidence intervals suggested that important differences were excluded. We conclude that at the doses used in this trial there were no important differences between captopril and atenolol in their effects on quality of life.


The haemodynamics of obesity: a theoretical analysis. Obesity and hypertension are frequently associated with one another, and changes in body weight are usually accompanied by consensual changes in blood pressure. To examine formally the haemodynamic conditions required for a weight gain to cause a rise in blood pressure in the resting state. The relevant equations were developed, and then used in a simulation to predict blood pressure and peripheral vascular resistance in response to changes in body weight. The model was tested on data taken from published reports. Furthermore, the impact of the composition of the excess weight, i.e. fat versus muscle tissue, on the haemodynamics of obesity was incorporated into the simulation by using a range of tissue-specific blood flow rates taken from published reports. The key determinant of weight-induced increases in blood pressure is a disproportional increase in cardiac output, which is not fully accounted for by the haemodynamic contribution of new tissues. An increase in muscle tissue relative to fat in newly gained weight is a further factor in the blood pressure increase arising from any given level of excess weight. The disproportional increase in cardiac output in obesity is best attributed to stimulation of cardiac dynamics by the adrenergic nervous system. The low resistance of adipose tissue relative to muscle may be the physiologic basis for the dominance of fat-free mass in multiple regression analyses of blood pressure determinants in the population; it further suggests that in the obese hypertensive, fat tissue may provide protection against cardiovascular disease by limiting increases in total peripheral resistance.


[Cellular or atypical mesoblastic nephroma of partially cystic form. Apropos of a case in a 27 year old woman]. A case of partially cystic atypical or cellular variant of mesoblastic nephroma in a 27 year old woman is reported herein. Some of the cysts were like those seen in a cystic nephroma (solitary multilocular cyst) and others resembled ectasic vascular channels. Nephrectomy was the only treatment used. Since her operation our patient has become pregnant and given birth to a healthy daughter. The mother is well and free of disease more than 18 months after nephrectomy. We have only found 7 other reports of adult mesoblastic nephroma. Ours in the first published case of the cellular or atypical variant of mesoblastic nephroma in an adult patient. Difficulty in diagnosis and treatment particular to this type of tumor is stressed upon with reference to pediatric and adult cases described in the literature.


[Effect of cyclic AMP and adenosine on hCG transcription by trophoblast cells]. It is well established that cAMP stimulates secretion of both alpha and beta subunits of hCG by trophoblasts. However, the molecular mechanism behind this phenomenon, especially that of beta subunit stimulation is still obscure. Moreover, while adenosine is known to play a pivotal role in the message transduction system of various cells, its effect on trophoblasts is not known. We therefore tried to answer these questions and found that cAMP stimulates hCG beta secretion by increasing transcription of its mRNA. However, no discrete cAMP responsive element like that seen in the hCG alpha gene seemed to be present in hCG beta transcriptional regulatory elements. It was assumed that hCG beta transcription is augmented indirectly by numerous trans acting elements which respond to cAMP. We have also shown that adenosine is a potent stimulator of hCG production. Its effect was synergistic with cAMP. It was therefore assumed that adenosine and cAMP are likely to act through different pathways.


[The significance of determination of human herpesvirus-6 (HHV-6) antibody during pregnancy]. HHV-6 which was detected in peripheral blood lymphocytes of lymphocytic patients is now said to be an agent causing exanthema subitum. We investigated the titer of HHV-6 antibody in 100 primigravida and 30 patients with spontaneous abortion by means of the indirect immunofluorescence method (IF method). The positive incidence in which the titer of HHV-6 antibody was more than 10 times in 100 primigravida was 82%. That of HHV-6 IgM antibody was 3% in those who had no complication during pregnancy. This supposes that there may be some women who were infected or were reactivated while they were pregnant. The positive incidence in 30 spontaneous abortion patients was 90%. 3 of them had IgM antibody to HHV-6, and HHV-6 antigen was detected on the epitherium of the chorionic villi by the IF method with HHV-6 monoclonal antibody in two of them. This suggests that the infection or reactivation of HHV-6 may be one of the causes of spontaneous abortions.


[A report on two siblings with vitamin D resistant rickets treated with 1 alpha-hydroxy-vitamin D3 until completion of the growth]. The treatment of vitamin D resistant rickets still raises much controversy. This report describes out experiences on 1 alpha OHD3 treatment in two siblings from childhood till completion of the growth. The treatment began from age 3 and 8 in these two patients respectively. The daily doses were necessary to maintain the healing and increased during the growth period so that the doses per body weight were virtually constant at 1 microgram/kg/day. Under this regimen, the improvement of rickets was recognized in the radiogram, and the lower leg deformity corrected. In one case the bone biopsy confirmed the improvement of disturbed calcification. The final height was 152.5 cm (-0.8 SD) and 149 cm (-1.4 SD) respectively suggesting the effectiveness of 1 alpha OHD3 on the growth promotion. Around the closure of growth plate, the regimen induced hypercalcemia. Thereafter the doses were reduced to one fourth or one fifth of those in the growth period.


High ulnar nerve palsy caused by the arcade of Struthers. A case of ulnar nerve entrapment neuropathy caused by the arcade of Struthers is reported. Nerve conduction studies showed a complete block and surgical decompression was successful.


Nerve conduction studies after treatment for carpal tunnel syndrome. 125 cases of carpal tunnel syndrome confirmed electrophysiologically were the subject of longitudinal nerve conduction studies to assess spontaneous improvement and effect of treatment. 36 cases showed a slowly progressive deterioration which became statistically significant only on lengthy follow-up; analysis of interval tests in these cases revealed that definite improvement or rapid worsening can occur in the interim. The 56 cases studied after local corticosteroid injections showed a statistically significant improvement at one month followed by an overall progressive return to the previous abnormal values in six to 12 months, indicating only slight and temporary alteration in the natural progression of the conduction deficit. The 33 cases which underwent surgical release of the median nerve were shown to have obvious and often rapid improvement, which was sustained for at least one year after surgery.


Pre-operative factors and treatment outcome following carpal tunnel release. The outcome of carpal tunnel release was evaluated retrospectively in 60 hands of 53 patients followed for six to 33 months (median ten months). Outcome was considered good in 27% (pain, weakness, and numbness were essentially resolved); fair in 42% (most of the symptoms improved); and poor in 32% (symptoms persisted or worsened). Patients whose pre-operative work activity was considered physically strenuous were associated with a slightly but significantly poorer outcome (60% good or fair) compared to those in light work or with no employment (89% good or fair). Proportionately fewer patients returned to their original work when they previously engaged in strenuous activity, ranging from 27% for those using air guns to 80% in light work. It appears that the highest chance of a poor outcome from carpal tunnel release occurs in patients who have either associated symptoms of thoracic outlet syndrome or physically strenuous work activities.


Intravitreal ganciclovir pharmacokinetics in rabbits and man. Cytomegalovirus (CMV) retinitis occurs in immunocompromised patients and can be treated by repeated intravenous or intravitreal injections of ganciclovir (GCV) or foscarnet. Due to toxicity and complications these modalities are not ideal. The development of alternative administration routes is hindered by a lack of pharmacokinetic data. Devices giving pseudo zero order release of GCV were implanted intravitreally first in rabbits and then in patients with AIDS associated CMV retinitis as part of a Phase I clinical trial. Steady state intravitreal GCV levels were obtained immediately after death and the elimination rate constants were calculated assuming first order pharmacokinetics. Normalizing for retinal surface area, distribution volume and anatomic volume, the retinal elimination rate constants were calculated. These were found to be 0.017 cm-2hr-1 in rabbits and 0.015 cm-2hr-1 in man. This indicates that the rabbit eye is a good model for studying intravitreal pharmacokinetics of ganciclovir and suggests a common elimination mechanism which may be trans-retinal.


Effects of steroids and immunosuppressive drugs on endotoxin-uveitis in rabbits. Anti-inflammatory actions of dexamethasone (DEXA), Cyclosporin A (CSA) and Rapamycin (RAPA) were assessed on uveitis induced by intravitreal E-coli Endotoxin (100ng) in rabbits at 24 hrs. In this model, endotoxin caused a breakdown of the blood-aqueous barrier (BAB) and polymorphonuclear neutrophils (PMN) infiltration into the aqueous humor (AH) and iris-ciliary body (ICB). Intramuscular (I.M.) DEXA (2mg/kg) but not topical DEXA (0.1% 6 x daily) inhibited AH leukocytes and protein level. However, both routes caused an inhibition of AH Prostaglandin E2 (PGE2) and Leukotriene B4 (LTB4). In the ICB, I.M. DEXA significantly inhibited PGE2 synthesis and myeloperoxidase (MPO) activity. I.M. CSA (25mg/kg) and I.M. RAPA (10mg/kg) inhibited the AH leukocytes and protein content and MPO activity in the ICB. RAPA also inhibited AH protein and eicosanoid (except AH LTB4) levels in both the AH and ICB. Interestingly, castor oil, a vehicle of CSA, also inhibited AH leukocytes and the release of PGE2 into AH and from ICB. In summary, systemic administration of DEXA and other immunosuppressive drugs CSA and RAPA significantly inhibited endotoxin-induced uveitis in rabbits.


Elevation of cytosolic free calcium in cultured ciliary epithelial cells by histamine: effects of verapamil and staurosporine. Treatment with histamine (10(-4) M) of cultured non-pigmented human ciliary epithelial cells led to a biphasic elevation of free intracellular calcium mediated by H1-receptors. The initial transient increase was due to Ca(2+)-release from intracellular calcium stores and could be blocked with a high concentration of verapamil (10(-4) M). The subsequent sustained elevation of cytoplasmic calcium caused by an influx of extracellular calcium was reduced by staurosporine (10(-7) M). We conclude that the sustained increase of cytoplasmic calcium by histamine may be partially mediated by activation of protein kinase C. Since depolarization of the cells had no effect on intracellular calcium, we conclude that typical voltage-operated calcium channels do not significantly influence intracellular calcium.


Altered stereoselectivity of cocaine and bupivacaine isomers in normal and batrachotoxin-modified Na+ channels. The inhibitory effects of local anesthetics (LAs) of cocaine and bupivacaine optical isomers on Na+ currents were studied in clonal GH3 cells under whole-cell patch clamp conditions. At holding potential of -100 mV, all four isomers inhibited peak Na+ currents when the cell was stimulated infrequently. The dose-response curves of this tonic block of peak Na+ currents by (-)/(+) cocaine and (-)/(+) bupivacaine were well fitted by the Langmuir isotherm, suggesting that one LA isomer blocked one Na+ channel. Each pair of isomers showed no greater than a twofold difference in stereoselectivity toward Na+ channels. Additional block of Na+ currents occurred when the cell was stimulated at 2 Hz. This use-dependent block was also observed in all four isomers, which again displayed little stereoselectivity. The voltage dependence of the use-dependent block produced by cocaine isomers did not overlap with the activation of Na+ channels but did overlap with the steady-state inactivation (h infinity), indicating that cocaine can bind directly to the inactivated state of Na+ channels before channel opening. In comparison, the peak batrachotoxin (BTX)-modified Na+ currents were little inhibited by cocaine and bupivacaine isomers. However, the maintained BTX-modified Na+ currents were highly sensitive toward the (-) form of cocaine and bupivacaine isomers during a prolonged depolarization. As a result, a profound time-dependent block of BTX-modified Na+ currents was evident in the presence of these LA isomers. The estimated values of the equilibrium dissociation constant (KD in micromolar) at +50 mV were 35.8, 661, 7.0, and 222 for (-)/(+) cocaine and (-)/(+) bupivacaine, respectively. Although chloramine-T (CT) also modified the fast inactivation of Na+ channels and gave rise to a maintained Na+ current during a prolonged depolarization, LA isomers showed no greater stereoselectivity in blocking this maintained current than in blocking the normal transient Na+ current. We conclude that (a) cocaine and bupivacaine isomers exhibit only weak stereoselectivity toward the LA receptor in normal and CT-treated Na+ channels, (b) BTX drastically modifies the configuration of the LA binding site so that the LA stereoselectivity of the open Na+ channels is altered by an order of magnitude, and (c) the (-) forms of cocaine and bupivacaine interact strongly with the open state of BTX-modified Na+ channels but only weakly, if at all, with the closed state. The last finding may explain why most LA drugs were reported to be less effective toward BTX-modified Na+ channels.(ABSTRACT TRUNCATED AT 400 WORDS)