The use of inhibitors of platelet activation or protease activity in platelet concentrates stored for transfusion. During storage of platelet concentrates the platelets show signs of activation, and extracellular protease activity becomes evident in the plasma. The consequences of platelet activation and plasma protease activity are potentially detrimental to the preservation of platelet function in vitro. The earlier use of prostaglandins during preparation of platelet concentrates to increase the harvest of platelets from whole blood did little to improve their shelf-life. Other compounds that sustain elevated cyclic AMP levels or that directly inhibit platelet agonists provide more effective inhibition of platelet activation during storage. Also, the inclusion of general or specific protease inhibitors appears to improve platelet preservation over extended storage periods. These studies demonstrate the possibility of prolonging the shelf-life of platelet concentrates stored at 22 degrees C through the addition of non-toxic formulations of inhibitors of platelet activation and protease activity.


Does the circadian pacemaker act through cyclic AMP to drive the melatonin rhythm in chick pineal cells? Cyclic AMP is a key regulator of melatonin production in the chick pineal gland. Agents that raise cyclic AMP levels (such as forskolin), or cyclic AMP analogues (such as 8-bromocyclic AMP), increase melatonin synthesis and release, whereas agents that lower cyclic AMP levels (including light) decrease melatonin synthesis and release. A circadian oscillator in these cells also raises and lowers melatonin output. We have been investigating the relationships between cyclic AMP and the circadian pacemaker in the regulation of melatonin production. In the chick pineal (unlike certain neuronal systems), the weight of the evidence indicates that cyclic AMP is not on an entrainment pathway to the circadian pacemaker. Instead, cyclic AMP appears to act downstream from the pacemaker. The pacemaker might itself act directly through cyclic AMP, regulating melatonin content by raising and lowering cyclic AMP levels. If this were the case, and if the effects of cyclic AMP levels on melatonin output are saturable (as they must be), then, in the face of such saturating levels of cyclic AMP, the pacemaker should no longer raise or lower melatonin output. To test this prediction, maximally effective concentrations of forskolin and 8-bromocyclic AMP were determined. Both agents markedly increased melatonin output. After 36 hr, cells were refractory to additional stimulation of melatonin output by addition of both agents together, or by higher concentrations of forskolin (although cyclic AMP levels could still be raised further). Nonetheless, the circadian pacemaker continued to raise and lower melatonin output: The rhythm persisted in the face of saturating levels of cyclic AMP. It is therefore suggested that the circadian pacemaker in chick pineal cells acts with, not through, cyclic AMP to regulate melatonin synthesis. Cyclic AMP and the pacemaker act synergistically to regulate serotonin N-acetyltransferase activity and the melatonin rhythm, with cyclic AMP mediating acute effects and amplitude regulation.


Connexin 43 expression in the mouse embryo: localization of transcripts within developmentally significant domains. The expression of the gap junction gene, Cx43, during mouse embryogenesis was characterized by an in situ hybridization analysis of mouse embryos from gestation days 4.5 to 12.5. This analysis revealed that Cx43 transcripts are differentially expressed as a function of development beginning at the blastocyst stage. In many regions of the embryo, Cx43 transcripts were found in discrete spatially restricted domains. This was observed in conjunction with the development of the brain, neural tube, prevertebra, limb, and various aspects of organogenesis. In some cases, the differential localization of Cx43 transcripts is associated with developmental processes mediated by inductive interactions, such as that of the eye, otic vesicle, kidney, and the branchial arches. In addition, in the 10.5 day embryo, Cx43 transcripts appear to be distributed as a gradient in regions spanning the midbrain/hindbrain junction, in the telencephalon, and in the limb mesenchyme. Surprisingly, our results also suggest that neural crest and sclerotomal cells, i.e., cells that are presumably migratory, express high levels of Cx43 transcripts. Overall, these results suggest that gap junctions encoded by Cx43 may play a role in various aspects of mouse development, possibly including relaying second messengers emanating from signal transduction pathways that mediate inductive interactions.


Chronic active Epstein-Barr virus infection in an adult. We report a rare adult case of chronic active Epstein-Barr virus (EBV) infection. A 54-year-old woman was admitted to our hospital with intermittent fever, weight loss, hepatosplenomegaly, pancytopenia and liver disturbance. In serological tests for EBV, anti-virus capsid antigen (VCA)-IgG antibody and anti-early antigen (EA)-IgG antibody were markedly elevated and anti-EBV nuclear antigen (EBNA) antibody was negative. EBV genome was detected in the bone marrow nucleated cells and peripheral lymphocytes by Southern blot hybridization. The patient developed left facial edema, bilateral breast tumor and pneumonia. She died one year after admission in spite of the administration of prednisolone, interferon and acyclovir.


Thyroid storm associated with probable subclinical hypoadrenocorticism in an elderly woman. A 73-year-old woman was admitted to the hospital for severe persistent vomiting with fever, drowsiness, and weight loss. Elevated serum levels of thyroid hormones and the presence of a consciousness disorder with fever and vomiting led to the diagnosis of thyroid storm. A low normal concentration of serum cortisol, urinary 17-hydroxycorticosteroids and an elevated plasma level of corticotropin suggest that an inadequate adrenal reserve have been involved in the pathogenesis of the thyroid storm in this patient. She responded to the administration of intravenous methimazole and oral supersaturated potassium iodide solution.


High performance liquid chromatographic analysis of six conjugated and unconjugated estrogens in serum. Six estrogens-estrone, equilin, estradiol, sodium estrone sulphate, sodium equilin sulphate and sodium 17-alpha-dihydroequilin sulphate in dog serum were successfully separated and quantified by a high performance liquid chromatographic method developed in our laboratories. The mobile phase was optimized by studying the effects of an organic modifier (acetonitrile) and an ion pairing reagent (tetrabutylammonium hydroxide). The serum sample work-up procedure was designed to recover both conjugated and unconjugated estrogens. The optimum method involved acetonitrile to precipitate serum proteins/peptides and extract estrogens. Residues were reconstituted in 50% acetonitrile in water for injection. The detection limits for these six estrogens via UV detection ranged from 0.5 to 5 ng on-column with a signal-to-noise ratio (S/N) of 10 for a 20 microL injection and via fluorescence 0.1 ng on-column for 17-beta-estradiol. Validation data are included for all six estrogens.


[Ion channels and drugs: what can be learned from their interactions?]. The study of the interaction between drugs and the sodium channel is important to understand the functioning of the channel at the molecular level. The affinity of a drug to bind depends on the channel's state: rested, activated or inactivated state. Since the probability of these three states depends on the membrane potential and the drug itself may carry a charge, there are two reasons for a modulation of the effect by the membrane potential. Drugs like lidocaine and tertiary amines bind preferentially to the inactivated state. Others like disopyramide and penticainide block only the open or activated channel and do not bind to the channel in the rested or inactivated state. The same rule has to be applied for the unblocking. The consequences of these differences are discussed.


99Tcm-Technegas and krypton-81m ventilation scintigraphy: a comparison in known respiratory disease. Krypton-81m gas, by virtue of its imaging characteristics, is often considered the "gold standard" for ventilation scintigraphy but its use is restricted by its high cost and limited availability. The new radiopharmaceutical 99Tcm-Technegas, a suspension of ultrafine technetium-99m labelled carbon particles, produces high-quality images of ventilation and has the advantage of continuous availability. As part of our evaluation of Technegas the two were compared in 40 patients with a variety of established respiratory diseases. Disparities were seen in five patients in five diagnostic groups and may be a consequence of the differing physical properties of the two agents and the different inhalation techniques used. In addition two interesting features were noted on the Technegas images. (1) Hot spots were seen in 50% of patients, particularly in those with a degree of airways obstruction; (2) preferential basal deposition of activity was seen in 30%, particularly in patients with idiopathic pulmonary fibrosis. Both features were significantly associated with parameters of pulmonary function indicating obstructive lung disease in the former case and restrictive lung disease in the latter.


Late follow-up of lung function after whole lung irradiation for Wilms' tumour. Radiotherapy is known to have acute and long term deleterious effects on lung tissue. However, pulmonary irradiation is an established treatment in advanced childhood tumours with pulmonary metastases not responsive to chemotherapy. In this study eight patients with Wilms' tumours and lung metastases treated with whole lung irradiation (1200-1837 cGy) and chemotherapy were reassessed clinically, radiologically and with lung function tests 6-26 years after radiotherapy. One patient was breathless after mild exertion, four after strenuous exercise and three were asymptomatic. Clinically all had small chests and four of five females had underdeveloped breasts. A chest radiograph showed clear lung fields in all cases. Lung volumes, especially total lung capacity (TLC) and vital capacity (VC), were decreased when compared with predicted values for age and height. However, gas transfer per unit lung volume (KCO) was normal. This study suggests that pulmonary irradiation in childhood results primarily in underdevelopment of the thorax and that diffuse interstitial lung fibrosis is not a significant feature at this dose level.


Interaction between mitogens upon intracellular Ca2+ pools in murine fibroblasts. A comparison of the effect of platelet-derived growth factor (PDGF) and bombesin on intracellular Ca2+ stores was carried out in Swiss 3T3 cells loaded with Fura-2. It was found that the tumor promoter thapsigargin (Tg) almost completely inhibited both the PDGF- and the bombesin-induced intracellular Ca2+ concentration ([Ca2+]i) rise, indicating that the two mitogens mobilize Ca2+ from intracellular pool(s) sensitive to the tumor promoter. It was also found that pre-treatment with PDGF almost totally and persistently (up to at least 30 min) inhibited the bombesin-, Tg- and ionomycin-induced rise in [Ca2+]i, whereas pre-treatment with bombesin had only a partial inhibitory effect on the PDGF, Tg and ionomycin [Ca2+]i response, both in the absence and in the presence of external Ca2+. On the other hand, vasopressin and bradykinin, which also stimulate hydrolysis of phosphoinositides in these cells, did not affect the [Ca2+]i response induced by the same agents. These results indicate that, despite the poor production of inositol 1,4,5-trisphosphate (InsP3), PDGF was capable of totally discharging and maintaining discharged the InsP3-sensitive stores of intracellular Ca2+, regardless of whether extracellular Ca2+ was present in the medium. Bombesin only partially caused this effect. On the contrary, bradykinin and vasopressin, after releasing intracellular Ca2+ allowed an almost total refilling of the pools. It is interesting to note that, at variance with PDGF and bombesin, neither bradykinin nor vasopressin are able to induce a mitogenic response in Swiss 3T3 cells.


Characterization of the energy-dependent, mating factor-activated Ca2+ influx in Saccharomyces cerevisiae. The yeast mating pheromones, a and alpha factors, bind to specific G protein-coupled receptors in haploid cells and bring about both growth arrest in the early G1 phase of the cell cycle and differentiation into mating capable cells. This induces an increase in Ca2+ influx leading to elevated intracellular calcium concentrations, which has been shown to be essential for subsequent downstream events and the mating process itself [1]. We have characterized the alpha factor induced increase in cellular Ca2+ in wild type S. cerevisiae and in the temperature-sensitive cell division cycle mutants cdc7 and cdc28 which are growth-arrested at the G0-G1 border at the nonpermissive temperature. We observed a 2-4 fold increase in the initial velocity of Ca2+ influx in alpha factor-treated wild-type cells and in cdc7 and cdc28 cells grown at the nonpermissive temperature. Calcium influx was energy dependent, inhibited by membrane depolarization and slightly increased by hyperpolarization. Furthermore, Ca2+ influx was sensitive to both divalent and trivalent cations, but was unaffected by nifedipine and verapamil. These data demonstrate that budding yeast possesses a regulated Ca2+ transport mechanism, the activation of which is dependent upon exit out of the cell cycle and growth cessation. This transport mechanism has many similarities to that observed in mitogen-stimulated mammalian cells.


Ryanodine-affinity chromatography purifies 106 kD Ca2+ release channels from skeletal and cardiac sarcoplasmic reticulum. A 106 kD protein was isolated from skeletal sarcoplasmic reticulum (SR) vesicles and shown to have the properties of SR Ca2+ release channels, including blockade by 5 nM ryanodine. In view of extensive reports that the ryanodine-receptor complex consists of four 565 kD junctional feet proteins (JFPs) and is the 'physiological' Ca2+ release channel, we prepared ryanodine-affinity columns to isolate its receptor site(s). Conditions known to maximize the association and dissociation of ryanodine to SR proteins were respectively used to link, then elute, the receptor(s) from ryanodine-affinity columns. The method purified a protein at about 100 kD from both rabbit skeletal and canine cardiac SR vesicles. The skeletal and cardiac proteins isolated by ryanodine-affinity chromatography were identified as the low molecular weight Ca2+ release channel through their antigenic reaction with an anti-106 kD monoclonal antibody. Upon reconstitution in planar bilayers, both skeletal and cardiac proteins revealed the presence of functional SR Ca2+ release channels. Surprisingly, ryanodine-affinity columns did not retain JFPs but purified 106 kD Ca2+ release channels which are a minor component (0.1-0.3%) of SR proteins.


Mechanisms of perturbation of erythrocyte calcium homeostasis in favism. Favism is an acute hemolytic anemia triggered by ingestion of fava beans in genetically susceptible subjects with severe deficiency of glucose-6-phosphate dehydrogenase (G6PD) activity. Erythrocytes from 10 favic patients had constantly and markedly increased calcium levels, as compared with values detected in 4 asymptomatic G6PD-deficient controls. Correspondingly, the calcium permeability of erythrocytes, estimated as the fraction of intracellular calcium exchangeable with externally added 45Ca2+, was invariably enhanced in favism and returned to normal patterns after several months from the acute hemolytic crisis. In favic patients, the levels of erythrocyte calcium ATPase activities showed wide variability, ranging from 2.0-12.9 mumol Pi/ml RBC/h, while control values in asymptomatic G6PD-deficient subjects were 10.62 +/- 2.03 mumol Pi/ml RBC/h. Analysis of the calcium ATPase in situ in erythrocyte membranes from favic patients showed the same molecular mass of 134 kD as observed in the control subjects. Exposure of G6PD-deficient erythrocytes in vitro to autoxidizing divicine, a pyrimidine aglycone strongly implicated in the pathogenesis of favism which leads to late accumulation of intracellular calcium, caused: (i) a marked inactivation of calcium ATPase, without changes in the molecular mass of 134 kD; and (ii) the concomitant loss of spectrin, band 3 and band 4.1, all known substrates of the calcium activated procalpain-calpain proteolytic system. Thus, the increased intraerythrocytic calcium apparently results in the degradation of calcium ATPase observed in some favic patients. It is proposed that both enhanced calcium permeability and a calcium-stimulated degradation of the calcium pump are the mechanisms responsible for the perturbation of erythrocyte calcium homeostasis in favism.


Inhibition of the human erythrocyte calcium pump by dimethyl sulfoxide. The action of dimethyl sulfoxide on the human red cell Ca2+ pump was studied in inside-out vesicles. In a high-K+ medium at pH 7.6, the organic solvent inhibited both Ca2+ transport and ATP hydrolysis. Half-maximal effect was obtained with about 2% (v/v). At or below 10% dimethyl sulfoxide, the inhibition was overcome by adding inorganic phosphate or oxalate. In the absence of organic solvent, Ca2+ efflux from Ca(2+)-loaded vesicles consisted of a slow and a fast component whilst in its presence, there appears additionally a leakage component. The size of the latter depended markedly on dimethyl sulfoxide concentration, being about 3% at that level where Ca2+ uptake was half-maximally inhibited. ATP hydrolysis was more sensitive to dimethyl sulfoxide (10%) when free Ca2+ was increased within the millimolar level than when it was raised within the micromolar range. On the other hand, raising Ca2+ with organic solvent greatly stimulated ATP synthesis through ATP-Pi exchange, without reaching saturation. The results suggest that dimethyl sulfoxide blocks the red cell Ca2+ pump by increasing the affinity of the Ca2+ translocating site at the releasing step. They also show that at high concentrations, this solvent increases Ca2+ permeability.


Mineralocorticoid excess and inhibition of 11 beta-hydroxysteroid dehydrogenase in patients with ectopic ACTH syndrome. 11 beta-Hydroxysteroid dehydrogenase protects renal mineralocorticoid receptors from cortisol by converting cortisol to inactive cortisone. We hypothesize that 11 beta-dehydrogenase is inhibited by ACTH, providing a mechanism whereby cortisol induces hypokalaemic alkalosis in ectopic ACTH syndrome. The principal sources of plasma cortisone were assessed by selective venous catheterization with measurement of cortisol and cortisone by radioimmunoassays. The effect of ACTH on peripheral plasma cortisol/cortisone ratio was assessed in healthy volunteers during circadian rhythm, insulin induced hypoglycaemia, and infusions with exogenous ACTH or cortisol. In patients with Cushing's syndrome plasma cortisol/cortisone ratios were related to plasma potassium, corticosterone, and 11-deoxycorticosterone concentrations. Catheterization was performed in 24 patients with valvular or ischaemic heart disease. Cushing's syndrome patients included: 15 with pituitary adenoma; two with adrenal adenoma; and nine with ectopic ACTH secretion. Plasma cortisol/cortisone ratios were low in renal vein and high in hepatic vein. In healthy volunteers plasma cortisone increased during cortisol infusion but did not change with increases in endogenous or exogenous ACTH. Plasma cortisol/cortisone ratios were higher in ectopic ACTH syndrome than in other forms of Cushing's syndrome. However, the cortisol/cortisone ratio was no better a predictor of hypokalaemia than the levels of 11-deoxycorticosterone or corticosterone. Peripheral conversion of cortisol to cortisone occurs mainly in the kidney and is inhibited by ACTH. In ectopic ACTH syndrome the characteristic mineralocorticoid excess can be accounted for by a combination of increased secretion of cortisol, corticosterone and of 11-deoxycorticosterone and decreased inactivation of cortisol and corticosterone by 11 beta-dehydrogenase.


In vitro activity of sparfloxacin and other antimicrobial agents against genital pathogens. The in vitro activity of sparfloxacin was determined for 60 strains of Neisseria gonorrhoeae, 15 strains of Chlamydia trachomatis and 40 strains each of Gardnerella vaginalis, Mycoplasma hominis, and Ureaplasma urealyticum and compared with those of ampicillin, azithromycin, clarithromycin, erythromycin, ofloxacin, temafloxacin and tetracycline. Sparfloxacin was active against all the strains studied and appeared to be the most potent quinolone tested. Sparfloxacin had the lowest MICs against N. gonorrhoeae (MICs 0.002-0.06 micrograms/ml). Its MICs against C. trachomatis (0.03-0.06 micrograms/ml) were higher than those of clarithromycin but lower than those of the other antimicrobial agents. Sparfloxacin was particularly active against tetracycline-susceptible as well as resistant strains of M. hominis (MICs, 0.06 micrograms/ml) and U. urealyticum (MICs 0.125-1 micrograms/ml). Because of this in vitro activity and its tissue distribution, sparfloxacin might be a valuable therapeutic agent for treating major bacterial sexually transmitted diseases.


The comparative activity of fleroxacin, three other quinolones and eight unrelated antimicrobial agents. The in vitro activity of fleroxacin, a new trifluorinated quinolone was evaluated against 432 bacterial isolates. Fleroxacin was 1- to 2-fold less active than ciprofloxacin and at least as active as ofloxacin and lomefloxacin against most members of the family Enterobacteriaceae. The MICs of fleroxacin for 90% of strains tested (MIC90) were < or = 0.25 micrograms/ml against all isolates of Enterobacteriaceae except Citrobacter freundii (MIC90, 4 micrograms/ml) and Serratia marcescens (MIC90, 2 micrograms/ml). Fleroxacin was as active as ciprofloxacin, ofloxacin and lomefloxacin against Pseudomonas spp, (MIC90 for all quinolones tested were > 8 micrograms/ml). Acinetobacter and Haemophilus influenzae were very susceptible to fleroxacin; however fleroxacin was 1-fold less active than lomefloxacin against Acinetobacter and at least 1-fold less active than ciprofloxacin or ofloxacin against H. influenzae. Methicillin-susceptible and -resistant strains of Staphylococcus epidermidis and methicillin-susceptible strains of S. aureus were very susceptible to fleroxacin, with an MIC90 < or = 1 microgram/ml (range 0.5-1 microgram/ml). Methicillin-resistant S. aureus and Staphylococcus spp. other than aureus and epidermidis were not susceptible to fleroxacin (MIC90 > 8 micrograms/ml). In addition, fleroxacin as well as ciprofloxacin, ofloxacin and lomefloxacin were inactive against Enterococcus spp. (MIC90 > 8 micrograms/ml). Streptococcus pneumoniae and S. pyogenes were resistant to both fleroxacin and lomefloxacin but were very susceptible to ciprofloxacin and ofloxacin. These results suggest that fleroxacin represents a valid therapeutic option in the treatment of infections caused by most Enterobacteriaceae and some species of staphylococcus.


In vitro susceptibility of 245 yeast isolates to amphotericin B, 5-fluorocytosine, ketoconazole, fluconazole and itraconazole. The in vitro activity of amphotericin B, 5-fluorocytosine, ketoconazole, fluconazole and itraconazole was tested against 245 yeast strains isolated from clinical specimens (68 Candida albicans, 74 Candida tropicalis, 43 Candida krusei, 28 Candida glabrata, 19 Candida parapsilosis, 8 Candida lusitaniae and 5 Candida guilliermondii). An agar dilution method was employed to carry out testing. Minimal inhibitory concentrations to restrain 90% of isolate growth (MIC90) ranged from 0.12 to 2 mg/l for amphotericin B and for 5-fluorocytosine, from 0.03 to 8 mg/l for ketoconazole, from 0.05 to 50 mg/l for itraconazole and from 0.1 to > 100 mg/l for fluconazole. Among the azole derivatives, the most active was ketoconazole, followed by itraconazole. Only 1 strain of C. albicans was resistant to amphotericin B (MIC > 4 mg/l). Both C. tropicalis and C. krusei responded poorly to fluconazole and the former to itraconazole as well. The species most susceptible to the antifungal agents tested was C. glabrata and the most resistant were C. tropicalis and C. krusei.


[Surgical treatment of 1450 patients with hepatocellular carcinoma]. An analysis of 1450 patients with pathologically and surgically proved hepatocellular carcinoma (HCC) revealed a steadily increased 5-year survival rate of 2.8% (1958-1968), 10.5% (1969-1979), and 36.6% (1980-1990), owing to increased number of cases with small HCC (0.9%, 9.9%, 25.2%, respectively), more and more patients undergoing tumor resection (0, 14, 59) and preoperative tumor bulk reduction for otherwise unresectable HCC (0, 1, 33 cases). Small HCC resection (n = 250) resulted in a high 5-year survival rate as compared to non-small HCC resection (n = 491) (66.3% versus 31.2%). The 5-year survival of 73 patients undergoing tumor re-resection was 40.6%, and the 5-year survival of 34 patients having second stage resection was as high as that of those having small HCC resection (62.0%). 125 patients survived more than 5 years; of these 65 underwent small HCC resection, and 43 non-small HCC resection. Thus, early resection, re-resection for subclinical recurrence, and second stage resection for originally unresectable HCC play an important role in improving prognosis of HCC.


[Hepatectomy under continuous normothermic interruption of hepatic blood flow in noncirrhotic patients]. Continuous normothermic interruption of hepatic blood flow for 20 minutes or longer was applied in 37 non-cirrhotic patients, including 7 having emergency operation and 30 elective operation. Hepatic blood inflow was interrupted for 40 to 47 minutes in 4 patients, 30 to 39 minutes in 7, 25 to 29 minutes in 14, and 20 to 24 minutes in 12. Postoperatively, 1 patient died of recurrent hemobilia, which was not related to intraoperative vascular interruption. Two patients had mild jaundice for 1 week after right trilobectomy, while the remaining 34 patients recovered uneventfully with no clinical hepatic insufficiency. This method is considered simple, safe, and acceptable for hepatic resection in non-cirrhotic patients.


[Small hepatocellular carcinoma. Diagnosis and treatment of 40 cases]. In 40 patients with small hepatocellular carcinoma, the lesions were less than 5 cm in diameter, with a mean diameter of 3.4 +/- 1.1 cm. The smallest was 1.1 x 1.3 cm. 39 patients were first detected by ultrasonic scanning, and one was discovered during emergency operation because of spontaneous tumor rupture. 36 patients were treated by means of hepatic resection. One (2.7%) died within 30 days after operation. Four patients with severe hepatic cirrhosis and significant prolongation of prothrombin time (4 seconds over control) were subjected to selective transcatheter arterial embolization combined with transcatheter arterial infusion of chemotherapeutic agent. The diagnosis and treatment of small hepatocellular carcinoma was discussed.


[Cryosurgery for primary hepatic cancer of 87 patients]. From november 1973 to June 1991, cryosurgery with liquid nitrogen was performed on 87 patients with primary liver cancer (PLC). Of these, 27 patients was of stage I (31.0%), 52 in stage II (59.8%), and 8 stage III (9.2%). There were 30 patients with PLC of < or = 5 cm (34.5%). Liver cirrhosis was found in 73 patients (83.9%). In the beginning, plate-like cryoprobes and thermocouples were used to monitor the frozen area. Later on we designed single- and multiple-needle cryoprobes for freezing tumors deeply into the hepatic parenchyma and intraoperative ultrasound was used to monitor hepatic cryolesions. The 1-year, 3-year, and 5-year survival rates were 60.5%, 32.0%, and 20.2%, respectively. Among the 30 patients with PLC of < or = 5 cm, the 1-year, 3-year, and 5-year survival rates were 92.6%, 66.6%, and 50.8%, respectively. There were no operative mortality and complications such as rupture of the tumor, delayed bleeding, and bile leakage. These results indicate that cryosurgery is a safe and effective local treatment for unresectable PLC.


[Cisplatin (CDDP) microcapsules in transcatheter hepatic arterial chemoembolization of primary liver cancer]. Hepatic arterial transcatheter chemoembolization with CDDP microcapsules (CDDP mc) was carried out in 19 patients with primary liver cancer (PLC). Tumor regression greater than 50% in diameter after therapy occurred in 58% of the patients, which was statistically significant as compared with another 22 PLC patients given conventional hepatic arterial chemoembolization (58% vs 23%, P < 0.01). Histological examination revealed that 4 of the five tumor specimens resected after the treatment showed complete tumor necrosis. The CDDP concentration was kept at a high level in the tumor region for a long period of time while peripheral drug level was low. Hepatic angiography showed that CDDPmc mainly obliterated the peripheral tumor vascularity with neither establishment of collaterals nor recanalization of the embolized tumor vessels. No serious complications were noted in all patients. Our results demonstrated that CDDPmc is very useful for hepatic arterial embolization with prolonged chemotherapeutic and embolic effects on PLC.


[Transcatheter chemoembolization combined with hepatectomy for primary hepatic carcinoma. A clinical and pathological study]. Transcatheter chemoembolization (TCE) followed by laparotomy was performed in 12 patients with primary hepatic carcinoma (HCC). Each patient received TCE for 1-5 times with an interval of 4-6 weeks. The embolizing reagents included mitomycin C (20-40 mg), adriamycin (20-60 mg) and 40% lipiodol (2-10 ml). Results showed that the AFP level remarkably decreased even became normal and tumor size reduced. Partial hepatectomy was carried out in all 12 patients undergoing laparotomy except one with extensive abdominal metastasis. Histopathological study showed tissue necrosis and fibrosis at different stages in the surrounding liver parenchyma especially in the tumour area. Apart from sclerotic lesions, one or two daughter nodules were found in 2 patients. residual liver cancer was found in all of the 11 patients. Reexploration was made one year after liver resection in one patient for local recurrence and metastasis. The other two with right lung metastatic tumour 4-6 months after operation were given transcatheter pulmonary arterial embolization. We consider TCE as an effective adjuvant therapy to the combined treatment of liver carcinoma. It is suggested that TCE is indicated for unresectable HCCs, resectable HCCs with poor liver function, those associated with portal hypertension, and recurrent HCCs.


[Residual liver cancer after transcatheter hepatic arterial chemoembolization in patients with large primary hepatic carcinoma]. 15 cancer specimens resected after hepatic arterial chemoembolization (TCE) were studied pathologically. It was found that residual tumor tissue was present in three forms: failure of complete tumor necrosis; tumor emboli in small portal veins; and multiple cancer foci in adjacent liver parenchyma. It is believed that presenting residual tumor tissue after TCE may be attributed to one of the following conditions: low drug concentration in residual tumor tissue; the larger tumor size after TCE; more residual cancer tissue in ASCCL than in HCC, collaterals formed soon after hepatic arterial occlusion.


Calcium calmodulin dependent kinase II in cat visual cortex and its development. A monoclonal antibody against the alpha-subunit of calcium/calmodulin-dependent protein kinase II (CAM-K II) was used to visualize the kinase in developing kitten visual cortex. CAM-K II was first expressed in neurons of the deep cortical layers (V and VI) at postnatal day 1-4 and appeared in the remaining cortical layers within the first 2 weeks. The level of immunoreactivity declined in cells of layer V and upper layer VI at about 30-40 days of age. By postnatal day 90, the most densely labelled neurons were concentrated in cortical layers II, III, lower layer IV and in layer VI. This laminar pattern remained constant into adulthood. EM studies showed that the kinase was found in both pre- and postsynaptic locations. About twice as many immunopositive neurons were found in cortical layers II-IV and VI in young adult cats when geniculate input was removed by an unilateral thalamic lesion performed early in life. These results indicate that expression of CAM-K II is developmentally regulated in visual cortical neurons; the alteration of immunoreactivity after early LGN lesions suggests that the level of the kinase (or its alpha-subunit) is also regulated by cortical input.


Ultrastructural localization of peroxidase in atherosclerotic lesions of pigeons. Atherosclerotic lesions are known to have metabolic alterations which are associated with progressive lipid accumulation. Among the changes, lysosomal enzyme activity has been extensively characterized and at the ultrastructural level has been correlated with the amount of foam cell lipid. In a fashion paralleling lysosomal change, artery wall peroxidase activity is also altered during disease progression. The present study focuses upon the ultrastructural localization of peroxidase activity in atherosclerotic lesions of the aorta and coronary arteries from White Carneau pigeons fed a cholesterol-supplemented (0.3%) diet for 3 years. This resulted in fibrous lesions, rich in smooth muscle cells. The birds were necropsied by perfusion fixation, and peroxidase cytochemistry was carried out using the diaminobenzidine reaction. Peroxidase activity was found within endothelial cells and smooth muscle cells in both the media and intima, but cytochemically demonstrable activity was not found in macrophage foam cells. Peroxidase was localized within the nuclear envelope and endoplasmic reticulum, especially within cells that had lipid inclusions. The degree of peroxidase positivity varied within and among the arteries. In nonlesion regions of the aorta 20% of medial smooth muscle cells was peroxidase positive; the value for coronary artery smooth muscle cells was less. The peroxidase activity within aortic lesions was increased with 44% of intimal smooth muscle cells being positive. Notably, 85-90% of the lipid-containing intimal smooth muscle cells were positive. In contrast, intimal smooth muscle cells in the coronary artery lacked peroxidase reaction product, even in cells containing lipid. We conclude from these studies that aortic lesions contain a cytochemically differentiated subset of lipid-containing, peroxidase-positive smooth muscle cells; but coronary lesions lack a comparable subset of smooth muscle cells.


Tryptophan binding to nuclei of rat brain. Nuclei purified from whole rat brain specifically bind [3H]tryptophan ([3H]Trp) under in vitro conditions. Excess unlabeled Trp (10(-4) M) is an effective inhibitor of in vitro [3H]Trp binding to brain nuclei. Rats tube-fed L-tryptophan (Trp) (30 mg/100 g body wt) 30 min to 4 hr before killing revealed decreased specific binding of [3H]Trp to purified brain nuclei in vitro. By Scatchard analysis, the nuclei from whole brain appear to contain one binding site for [3H]Trp, and the KD is 263 nM. A number of Trp-related compounds, Trp metabolites, or other amino acids and their analogues were observed to compete for in vitro [3H]Trp binding to brain nuclei. The ability of Trp analogues, metabolites, and other cognate compounds to inhibit in vitro [3H]Trp binding to brain nuclei was evaluated and utilized to map the active site of Trp binding.


Immunogold silver staining method for light microscopic detection of peripheral blood lymphocyte surface antigens with monoclonal antibodies. Labelling of peripheral blood Lymphocytes surface antigens was carried out using the method of colloidal gold, enhanced with silver staining. Instead of PBS the minimal essential medium (MEM) according the Eagle, pH 7.2, was used rinsing of isolated lymphocytes. Visibility of positive reactions on lymphocytes at application of both mentioned media was the same. Positive reaction at demonstration of p24 BLV on cells acquired the from of black cap while the IgG expression was observed in the from of diffuse dispersion of colloidal gold on cells. Differences between the application of individual media were observed in the shape of peripheral lymphocytes in smears. Utilization of Eagle's MEM resulted in more uniform shapes and optically smooth surfaces when viewed under a light microscope.


Comparative study on effects of cytochalasins B and D on F-actin content in different cell lines and different culture conditions. Effects of cytochalasins on actin polymerization state in living cells were measured using fluorimetry of TRITC-phalloidin bound to F-actin. Normal (3T3) and tumour (SV-3T3, B16 melanoma, and Ehrlich ascites) cells were treated with cytochalasin B and cytochalasin D (1 microgram/ml). Three effects of cytochalasins were demonstrated--depolymerization of F-actin, promotion of polymerization, and redistribution of actin without change in polymerization state. Occurrence of a given effect was dependent on cell type, cell density, cytochalasin concentration and type. This indicates that cells from different lines, and even the same cells in different culture conditions may differ significantly in their state of actin polymerization, which we suppose is the cause of their different reactions to cytochalasins. Accordingly, caution should be taken in generalizing the results concerning the effect of cytochalasis on the polymerization state of actin.


[The effect of antimembrane testicular antibodies on Na+, K(+)-ATPase activity in the testicles of rats of different ages]. The effect of large and small doses of rabbit antibodies specific to plasma membranes of the rat testicle cells has been studied in the experiments on Wistar rats of three age groups (preadolescent--aged 20 days, puberal--aged 5-7 months and old--aged 24-26 months). It is stated that incubation of plasma membranes by IgG fraction isolated from antimembrane testicular serum (IgG-ATCSm) in a large dose (43 g of protein G per 125 g of protein of membrane fraction) caused statistically reliable inhibition of Na+, K(+)-ATPase activity in the membranes of testicle cells of puberal and old rats. Preadolescent rats exhibit only a tendency to decrease the activity of this enzyme. Incubation of plasma membranes of testicle cells in rats of different age by small doses of IgG-ATCSm (0.43 g of protein G per 125 g of membrane protein) induced a statistically reliable increase of Na+, K(+)-ATPase activity in puberal and old animals and its slight increase in preadolescent rats. The IgG fraction isolated from normal rabbit serum (IgG-NRS) exerted a less pronounced effect on Na+, K(+)-ATPase activity parallel with retention of a tendency to a decrease of activity under the influence of large doses of the drug and to an increase with introduction of small doses.


Faecal lipid excretion levels in normal Japanese females on an unrestricted diet and a fat-restricted diet measured by simultaneous analysis of faecal lipids. Faecal lipid excretion was determined in 16 females on an unrestricted diet and on a fat-restricted diet using a chromatographic method for the simultaneous analysis of faecal lipids. The fat-restricted diet reduced the total quantity of faeces and the amounts of fatty acids, neutral sterols and bile acids excreted were almost halved compared with when on an unrestricted diet. This indicates that dietary fat, fibre and cholesterol affect the amount of faecal bile acid, neutral sterol and fatty acid excretion. The amount of cholesterol/animal sterols excreted and the percentage of primary bile acids were, however, similar for both the fat-restricted and unrestricted diets.


[The adrenoreceptor regulation of ureteral contractile function in the guinea pig]. The effects of specific agonists and antagonists of adrenoceptors and inhibitors of cAMP phosphodiesterase on electrostimulated phasic contractions in the ureter of guinea pig were studied. It has been shown that there mainly excitatory alpha 1-adrenoceptors in this object, the density of beta-adrenoceptors is slight and functional alpha 2-adrenoceptors are probably absent. Some aspects of adrenergic regulation of the contractile function of guinea pig ureter are discussed.


Uncomplicated urinary tract infections: lomefloxacin versus trimethoprim/sulphamethoxazole. Data were collected from 14 French centres which participated in a randomized study to compare the safety and efficacy of 400 mg lomefloxacin taken orally once daily by 62 patients with 160/800 mg trimethoprim/sulphamethoxazole (TMP/SMX) taken orally twice daily by 64 patients with uncomplicated urinary tract infections. Most patients were infected with Escherichia coli at baseline (72.4% in the lomefloxacin group and 69.0% in the TMP/SMX group) and all patients were treated for 5 days. At 5-9 days post-treatment, lomefloxacin had eradicated the causative organism of infection in 100% of evaluable patients treated with lomefloxacin compared with 86.7% of those treated with TMP/SMX. At 4-6 weeks post-treatment, there were no marked differences in eradication rates between the two treatment groups: 83.3% and 80.0% for the lomefloxacin and TMP/SMX groups, respectively. Clinical cure rates showed no marked differences between treatment groups at 5-9 days or at 4-6 weeks post-treatment. At 5-9 days post-treatment, lomefloxacin achieved a clinical cure rate of 78.6% compared with 86.7% for TMP/SMX evaluable patients. At 4-6 weeks post-treatment, the clinical cure rates were 66.7% and 86.7% for the evaluable lomefloxacin- and TMP/SMX-treated patients, respectively. Both treatment regimens were well tolerated with a low incidence of adverse events. In conclusion, once-daily oral dosing with lomefloxacin is a safe and efficacious alternative to twice-daily dosing with TMP/SMX in the treatment of uncomplicated urinary tract infections.


Acceptability of Aujeszky's disease vaccines. Vaccines against Aujeszky's disease are not only used to prevent the clinical consequences of a field infection, but also to support eradication of the virus. The current Aujeszky's disease vaccines (ADV) protect against severe clinical signs of disease and they reduce but usually do not prevent virus multiplication and excretion or the establishment of latency after infection. Vaccines also limit virus multiplication after reactivation. The efficacy of vaccination is reduced by passively acquired maternal antibodies. The mechanisms that afford immunity to the virus are only poorly understood. No simple parameters for immunity are therefore available. The European Pharmacopoeia formulates requirements for inactivated and live ADV vaccines for parenteral use. The vaccines must be safe; they must not induce local or systemic reactions; they must not be transmitted to unvaccinated pigs; they must not be transmitted by semen and across the placenta; the attenuation must be irreversible (live vaccines); the inactivation must be complete (inactivated vaccines); they must prevent mortality and limit growth retardation after challenge infection; the vaccine must not contain contaminating micro-organisms and viruses. No requirements have been formulated with regard to reduction of excretion of challenge virus after experimental infection, efficacy in pigs with maternal antibodies, reproducibility of efficacy studies, reduction of virus transmission under field conditions, the presence of a serological marker, safety for other species, and safety and efficacy of intranasally administered vaccines. Future developments should be directed to the development and evaluation of ADV vaccines that are able to limit transmission of the virus.


Safety of infectious bursal disease vaccines: assessment of an acceptability threshold. This study was undertaken to check the safety of commercially available infectious bursal disease (IBD) vaccines in terms of bursal damage, and their immunodepressive effects as evaluated by testing the birds after vaccination for their response to Newcastle disease vaccination. Further requirements are proposed to establish a suitable safety standard.


[The characteristics of the effect of inert gases on tissue respiration in vivo]. Helium-oxygen and argon-oxygen gaseous mixtures have been studied for their effect on tissue respiration. It is shown that 1-hour exposition of animals induces an increase in the oxygen consumption of the liver tissue (more essential in the presence of helium in the gaseous mixture).


[Genetic correlation between the arterial pressure response during emotional stress and the alpha1-adrenoreceptor concentration in brain regions]. Arterial blood pressure reactivity to the emotional stress and brain alpha 1-adrenoreceptors concentrations were studied in hypertensive (ISIAH strain) and normotensive (Wistar strain) rats and their F1 and F2 hybrids. Significant correlations between the stress-induced increase in the arterial blood pressure and the amount of alpha 1-adrenoreceptors in hypothalamus (+0.46) and medulla (+0.38) were found in the F2. This cosegregation may point to the significant role of genetically determined peculiarities of expression of alpha 1-adrenoreceptors in brain regions during pathogenesis of arterial hypertension in the ISIAH strain.


[Dermatoglyphic changes in some human hereditary disorders: syndactyly]. The dermatoglyphic hand prints from 19 patients with different types of syndactyly were analysed. It was shown that some digital triradii and palmar lines were missing and replaced by only one triradius with common radiants and one main palmar line in patients with syndactyly. With fingers fused incompletely so called zygodactylous triradius and the main Z line may appear, instead of or alongside with them. It is proposed that the position of local cell death in the interdigital spaces is determined by positional information which is expressed in the system of polar coordinates.


[Mobile genetic elements and quantitative characters in Drosophila: facts and hypotheses]. This review is dedicated to the comparison of the facts obtained and the proposed hypotheses, to the critical analysis of the situation arisen, and to the estimation of key propositions of the concept developed. The main point is that mobile genetic elements (MGEs) participate directly in expression, variability, selection and evolution of different quantitative characters. Genetic and selection data are considered, and hypotheses of random fixation, marker effect and direct participation of MGE patterns in expression and selection of quantitative characters are discussed. The consequences of temperature treatment are considered and hypotheses of masked selection and temperature induction of transpositions are discussed. The marker effects are shown to be non-sufficient to explain the properties of quantitative character radius incompletus system. The MGE patterns are important components of genetical system of determination of a quantitative character. MGEs modify, enhance the expression of neighbouring polygenes. Temperature effects could be explained by the influence of stress temperature treatment through the system of heat shock response on the capacity of MGEs to transcribe and transpose. The system of diversed MGE patterns in drosophila chromosomes could be believed to be universal genomic system of "soft" modification of the polygenic control of any limiting quantitative characters.


[Plasmid determined degradation of sulfo-aromatic compounds by Pseudomonas sp. BS1304]. Utilized sulfo-aromatic compounds of Pseudomonas sp. BS1304 were isolated. It was shown that the first step of conversion is desulfonation with following meta-cleavage of the substituted aromatic ring. At least two steps are controlled by the plasmid pBS1004 (120 kb), belonging to the IncP-9 incompatibility group. The degradative plasmid marked by Tn5-lac may be mobilized to P. putida, P. aeruginosa, P. mendocina, where a degradative phenotype is expressed.


[Interaction between the enhancers of yellow and the yellow gene in Drosophila melanogaster]. Earlier, mutations at six loci designated enhancers of yellow (e(y)1-6) were obtained in the system of prolonged instability, where transpositions of Stalker mobile element occur. These mutations decrease pigmentation of the cuticle in a number of alleles. In the present work, genetic analysis of the mechanism of action of e(y)1u1, e(y)3u1 and newly described e(y)7u1 mutations on the yellow locus expression was conducted. The data are obtained proving that the enhancers of yellow 1 and 3 control transcription of the yellow locus. At the same time, the interaction of here described gene e(y)7 with the yellow gene is likely to occur at the level of protein products.


Chromosome interactions in P-M dysgenic male recombination of Drosophila melanogaster. The frequency, distribution and structure of P elements on the second and third chromosomes of Texas 1, a wild-type inbred strain of Drosophila melanogaster, were investigated by in situ hybridization. These autosomes were isolated individually and used as P-element donors to study the frequency and distribution of male recombination events generated on recipient chromosomes which were originally devoid of P sequences. The P-element array of chromosome 2 was shown to generate higher male recombination frequencies on chromosome 3 than vice versa, despite having fewer P factors and fewer P elements in general. This is likely to be due to the presence and distribution of specific P-deletion derivatives, which vary in their ability to repress P mobility. The male recombination generated on recipient chromosomes is associated with the insertion of donated P sequences, but only in a small minority of cases could a novel P-element site be detected at, or near, the recombination breakpoint. The majority of such breakpoints appear to be associated either with unsuccessful P insertion, or with the action of P transposase attracted by P elements newly inserted elsewhere on the recipient chromosome. Recent evidence also suggests that a small proportion of the breakpoints may be associated with the action of P transposase alone. Male recombination breakpoints appear to be distributed effectively at random along the recipient autosomes, and their frequency of occurrence was shown to correlate with the physical length of DNA available between markers, as revealed by the polytene map distance.


Hydroxyl radical generation in the NADH/microsomal reduction of vanadate. ESR spin trapping measurements demonstrate generation of hydroxyl (.OH) radical from reduction of vanadate by rat liver microsomes/NADH without exogenous H2O2. Catalase decreases the .OH signal while increasing a vanadium (4+) signal. Addition of superoxide dismutase (SOD) or measurements under an argon atmosphere show decreased .OH radical production. The results suggest that during the one-electron vanadate reduction process by microsomes/NADH, molecular oxygen is reduced to H2O2, which then reacts with vanadium (4+) to generate .OH radical via a Fenton-like mechanism.


EPR spectra of DMPO spin adducts of superoxide and hydroxyl radicals in pyridine. Electron spin resonance spectroscopy and the spin trapping technique were used to study the formation of the superoxide radical in pyridine. 5,5-Dimethyl-1-pyrroline-N-oxide (DMPO) was employed as a trapping agent. Superoxide radical was generated using chemical (potassium superoxide) and photochemical methods with anthralin, benzanthrone, rose bengal, 1,8-dihydroxyanthraquinone and zinc tetraphenylporphyrine as photoactive pigments. Hyperfine coupling (hf) constants for DMPO/O2.- were determined to be aN = 12.36 G, a beta H = 9.85, G, a gamma H = 1.34 G. The aN and a beta H hf constants are in good agreement with values calculated from a previously determined relationship between hf constants and solvent acceptor number (Reszka et al., (1992) Free Radical Res. Commun., in press). When concentrated hydrogen peroxide was added to DMPO in pyridine a similar EPR spectrum was observed. It is suggested that in this case the DMPO/.O2H adduct is formed by nucleophilic addition of H2O2 to DMPO to give a hydroxylamine, followed by oxidation to the respective nitroxide. The EPR spectrum observed when tetrapropylammonium hydroxide and H2O2 were added to DMPO in pyridine had hf couplings aN = 13.53 G, a beta H = 11.38 G, a gamma H = 0.79 G and it was assigned to a DMPO/.OH adduct. This assignment was based on similarity of this spectrum to the one produced by UV photolysis of hydrogen peroxide and DMPO in aqueous solution and subsequent transfer to pyridine.


Possible reasons for differences in phototoxic potential of a 5 quinolone antibacterial agents: generation of toxic oxygen. The reason for the differences in phototoxic potential between the 5 quinolone antibacterial agents lomefloxacin, enoxacin, ciprofloxacin, ofloxacin and DR-3355 (the s-isomer of ofloxacin) in mice was investigated. Superoxide anion, hydrogen peroxide (H2O2), and bleaching of p-nitrosodimethylaniline (B-NDMA) were detected in quinolone solutions during irradiation with ultraviolet-A (UVA). Apparent levels of H2O2 and the B-NDMA per mole of quinolone paralled the phototoxic potentials in the mice. The N-NDMA induced by quinolones and UVA was inhibited partially by treatment with D-mannitol and dimethylsulfoxide, and also with diethylenetriamine-pentaaceticacid (DTPA), suggesting that Haber-Weiss and Fenton reactions occurred. UVA concentration-dependently increased the level of the B-NMDA in H2O2 solution and the swelling in the ear pretreated by intra-auricular injection of H2O2. Both augmentations were inhibited by DTPA or DMSO. The swelling induced by the 5 quinolones and UVA was completely inhibited by pretreatment with dimethylsulfoxide. Oxygen consumption was detectable during the photodegradation, and increased with time. These results showed that the phototoxic potentials of the 5 quinolones were probably related to the amounts of toxic oxygens generated in the target cells during irradiation.


Electrochemical sensors for direct reagentless measurement of superoxide production by human neutrophils. Electrochemical sensors based on immobilised cytochrome c or superoxide dismutase for the measurement of superoxide radical production by stimulated neutrophils are described. Cytochrome c was immobilised covalently at a surface-modified gold electrode and by passive adsorption to novel platinised activated carbon electrodes (PACE). The reoxidation of cytochrome c at the electrode surface upon reduction by superoxide was monitored using both xanthine/xanthine oxidase and stimulated neutrophils as sources of the free radical. In addition, bovine Cu/Zn superoxide dismutase was immobilised to PACE by passive adsorption and superoxide, generated by xanthine/xanthine oxidase, detected by oxidation of hydrogen peroxide produced by the enzymic dismutation of the superoxide radical. A biopsy needle probe electrode based on cytochrome c immobilised at PACE and suitable for continuous monitoring of free radical production was constructed and characterised.


[Importance of calcium ions and calcium antagonists in affective psychoses]. Apart from the eminent changes in the neurotransmitter systems of the central nervous system, a disturbance of the calcium ion concentration may be of significance in the pathophysiology of affective psychoses. The present paper deals with the contribution of calcium ions in the generation of affective psychoses and discusses the calcium antagonism as a new strategy in the treatment of the disease. The following topics will be described: 1. Disturbances of calcium metabolism in affective psychoses, 2. a comparison of lithium and carbamazepine effects, 3. clinical studies with organic calcium channel blockers in affective psychoses and 4. the role of calcium ions in affective psychoses.


[Anti-cyclic manifestation of asthma bronchiale and schizophrenic psychosis]. The interaction between the central nervous system and the immune system has been focus of recent research. Whereas cyclic AMP has been described as a link between atopic and affective disorders, a possible link between atopic disorders and schizophrenia has not yet been investigated. A 33-year-old patient showed episodes of allergic-asthmatic and paranoid-catatonic symptoms which appeared in an anticyclic pattern. Biological, clinical and therapeutic aspects are discussed in respect to the interaction between immunological and psychiatric disorders. Cyclic-AMP as a second-messenger is not specific for a certain cell species and might represent a possible link for integrated communication between the nervous and the immune system.


Transfection of SV40-transformed ataxia-telangiectasia fibroblasts with mouse DNA corrects hypersensitivity to neocarzinostatin and activates fibronectin gene expression. SV40-transformed ataxia-telangiectasia (SV40-AT) fibroblasts were cotransfected with a plasmid carrying the neomycin-resistance gene as well as DNA from primary mouse embryo fibroblasts. The transfected fibroblasts were seeded under selective conditions and neomycin-resistant (neor) colonies were obtained and tested for the effect of the carcinogen neocarzinostatin (NCS) on DNA synthesis. Whereas the primary A-T and SV40-transformed A-T fibroblasts did not respond to carcinogen treatment and continued to synthesize DNA on damaged templates, normal fibroblasts stopped DNA synthesis after NCS treatment. Among the neomycin-resistant colonies, cells of two colonies responded to NCS treatment by the cessation of DNA synthesis like normal fibroblasts. When DNA from such a colony was transfected into SV40-AT cells, four neor colonies were isolated of which one had regained the normal phenotype. This study provides the first clue that mouse DNA can partly correct the A-T genetic defect expressed in SV40-transformed fibroblasts. Two of the neor colonies with the corrected phenotype expressed a 3.5 kb fibronectin RNA that was detectable by a rat fibronectin DNA probe but not by the human fibronectin DNA probe containing the cell attachment sequence. The latter probe did not detect fibronectin mRNA in the SV40-AT cells but detected expression of the 8.6 kb fibronectin RNA in the two neor colonies of transfected SV40-AT fibroblasts in which the response to NCS was repressed. The results suggest that "correction" of the A-T gene defect in SV40-AT fibroblasts might be associated with regulation of human fibronectin gene(s) expression.


Colonic proliferation and colon cancer risk. A review of clinical studies. The purpose of this review is to discuss some of the recent hypotheses regarding the dietary etiology of colorectal cancer. We will review some of the epidemiological background and animal studies that were performed to test these hypotheses. The use of the rectal mucosal proliferation rate as measured by thymidine labelling index as an intermediate endpoint for clinical studies is discussed, including the results of testing these hypotheses using this marker. We believe that all hypotheses must eventually be tested in the human model and that carefully controlled, sequential analyses of each acceptable hypothesis is the most prudent road to understanding the environmental causes of colorectal cancer.


Specificity of antibody responses affected by extracorporeal immunoadsorption of plasma over columns of protein A silica. A relationship is described between the interaction of circulating immune complexes (CIC) from plasma with staphylococcal protein A immunoadsorption treatment columns and modulation of antibody responses related to the specific CIC. Eluates from the initial immunoadsorption columns used to treat a series of patients with breast adenocarcinoma, cancer chemotherapy-associated thrombotic thrombocytopenic purpura/hemolytic uremic syndrome (C-TTP/HUS), or immune thrombocytopenic purpura (ITP) were evaluated for disease-specific CIC containing Lex glycosphingolipid (Lex gl) adenocarcinoma-associated antigens or platelet autoantibody (anti-GPIIb/IIIa), together with the corresponding neutralizing antibody [anti-F(ab')2], and for nonspecific CIC containing cytomegalovirus (CMV) or herpes simplex virus type 1 (HSV-1) antigens. In addition, the levels of antibodies directed against CMV, HSV-1, Lex gl, and GPIIb/IIIa antigens, as well as anti-F(ab')2 antibodies, were compared in pretreatment and posttreatment serum samples. Columns used to treat breast adenocarcinoma patients contained only Lex gl CIC, and the only immunologic change observed after treatment was significant increases in anti-Lex gl antibodies in some patients. Columns used to treat C-TTP/HUS patients contained anti-GPIIb/IIIa-anti-F(ab')2 CIC, in addition to Lex gl CIC. After treatment, significant increases in anti-Lex gl and anti-F(ab')2 antibodies and significant decreases in anti-GPIIb/IIIa antibodies were observed in some patients. Columns used to treat ITP patients only exhibited anti-GPIIb/IIIa-anti-F(ab')2 CIC, and after treatment only decreases in anti-GPIIb/IIIa and increases in anti-F(ab')2 antibodies were observed in some patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Herpesviral Fc receptors and their relationship to the human Fc receptors. Nearly two decades ago, it was observed that cells infected with herpes simplex virus (HSV) acquired an IgG Fc binding activity. The properties of the viral Fc receptor (FcR) have now been characterized by several laboratories. The Fc binding activity appears on the surface of the infected cell prior to formation of progeny virions. The FcR induced by HSV has been identified as the HSV glycoprotein, gE. When HSV gE forms a complex with a second HSV glycoprotein, gI, the receptor binds IgG with higher affinity. Varicella-zoster virus (VZV), which is closely related to HSV, has also been shown to induce an FcR. Like the HSV FcR, the FcR specified by VZV possesses characteristics common to viral glycoproteins. VZV encodes two glycoproteins, gpI and gpIV, which are the homologs of HSV gE and gI. The VZV glycoproteins have many properties common to cell surface receptors, including O-linked glycans and phosphorylation sites. However, extensive computer-assisted analyses of the amino acid sequences of VZV gpI and gpIV did not uncover regions of homology to the human cellular Fc receptors for IgG.


Prevalence of herpesvirus, human T-lymphotropic virus type 1, and treponemal infections in Southeast Asian refugees. Sera obtained for treponemal serology (VDRL) from 193 Southeast Asian refugees representing five ethnic groups seen in a primary care clinic were examined for antibodies to human T-lymphotropic virus type 1 (HTLV-1), human herpes-virus-6 (HHV-6), Epstein-Barr virus (EBV), and cytomegalovirus (CMV). The seroprevalence was highest for EBV (99%), followed in decreasing order by CMV (95%), HHV-6 (26%), and HTLV-1 (0.6%). The VDRL was positive in 15% of patients. The highest seroprevalence to HHV-6 was noted in the Chinese (33%) and the lowest in the Laotian hilltribes, the Mien and Hmong (14%). Antibody to HHV-6 was most prevalent among patients under 20 and those between 60 and 69 years of age. Differences were not found among ethnic groups in the seroprevalence of HTLV-1, EBV, or CMV.


Use of antigen expressed in bacteria for detection of EBV-specific thymidine kinase antibodies in sera from patients with nasopharyngeal carcinoma. Two cDNA clones covering the N- and C-terminal portions of the EBV BXLF1 open reading frame were selected from a cDNA library derived from P3HR1 cells. The two clones were ligated, the N-terminal untranslated region truncated, and the product inserted into an E. coli expression vector, pET3CP*. The fusion protein was expressed under control of the T7 phage phi 10 gene promoter and shown to possess thymidine kinase activity. The protein was then used as an antigen to detect antibody reactivities in serum samples of nasopharyngeal carcinoma patients and healthy blood donors. Using a 1:400 dilution of serum samples in Western blot analyses, it was possible to differentiate the reactivities of serum IgA of NPC patients and healthy donors. The prevalence of positive reactivity to EBV TK in NPC was around 84%. The test was compared to others used for early diagnosis of NPC and was able to detect some patients who were negative in those tests.


Polymerase chain reaction assay for hepatitis C virus RNA using a single tube for reverse transcription and serial rounds of amplification with nested primer pairs. A procedure is described for the detection of hepatitis C virus (HCV) RNA in blood by means of the polymerase chain reaction (PCR) in which the reverse transcription step and two rounds of amplification are carried out in a single tube. This results in fewer manipulations, reduced risk of contamination, and economy of time. The procedures are generally applicable to other assays based on the PCR. We describe the preparation (from 100 microL serum) of test samples that remain stable for at least 6 days under specified conditions and an assay that employs nested primer pairs homologous to conserved sequences in the 5' noncoding region. The method was tested on 107 sera from the United States and Japan. Correlation with first-generation anti-HCV was 77%. Two sets of nested primer pairs homologous to sequences in the 5' noncoding region and one set based on structural region sequences showed differences in their reactivities with serum HCV RNA. The recommended single tube procedure specified a primer for reverse transcription that was conserved in all reported HCV genomes but absent from pestivirus genomic sequences. The effects of preanalytical factors on the detection of HCV RNA were studied. Qualitatively, there was no change in the HCV RNA-positivity of sera that were exposed to room temperature for 24 hours. Quantitative studies showed a decrease in titer in some specimens. Three cycles of freeze-thawing had no detectable effects on the titers of HCV RNA.


Superoxide anion production by leukocytes exposed to post-ischemic skeletal muscle. Superoxide anion (O2-) and polymorphonuclear leukocytes (PMNs) have been implicated in the genesis of skeletal muscle ischemia-reperfusion (I-R) injury, but the source of (O2-) has not been established. We studied PMNs as a potential source of O2- using a ferricytochrome reduction assay in 5 anesthetized dogs. Using a gracilis muscle model of I-R, 6 hours of ischemia was followed by 2 hours of reperfusion. The contralateral muscle served as control. Prior to ischemia and after 0.5 and 2.0 hours of reperfusion, PMNs were separated from the gracilis venous effluent of ischemic (I) and control (C) muscles. Central venous samples were also obtained prior to surgical preparation and after reperfusion. Assays for O2- were performed with and without zymosan (Z) activation. Results are expressed as nmol O2-/2 x 10(6) PMNs +/- SEM. Baseline production of O2- was 0.49 +/- 0.54 in central venous samples; Z increased the values to 6.77 +/- 2.13. After 2 hrs of reperfusion, central O2- was 1.57 +/- 0.75, which increased to 7.1 +/- 1.04 with Z. Gracilis venous samples O2- values with and without Z are reported in Table I. One way measures of analysis of variance showed no significant (p > 0.05) differences between samples. Our results demonstrate that PMNs are not the sole source of O2- in the pathophysiology of skeletal muscle I-R injury. PMN associated injury may be mediated by mechanisms other than O2- production.


Risk factors inducing the persistence of high-risk genital papillomaviruses in the normal cervix. To evaluate the risk factors associated with persistence of human papillomaviruses (HPV) types 16, 18, and 33 in the normal cervix, a prospective study was carried out in Belgium of 323 women without cytological evidence of cervical intraepithelial neoplasia. Demographic and clinical data were obtained by interview, and HPV DNA was assayed in cervical-swab specimens using the Fast Multiplex Polymerase Chain Reaction-based screening and confirmatory tests. A multivariable linear regression model was constructed using four well-known risk factors: the use of an oral contraceptive which is either triphasic, or monophasic and containing ethynylestradiol in association with either norethysterone, or levonorgestrel, or lynestrenol, or gestoden, or estrogenic and containing estriol (P = 8 x 10(-5)), a positive history of genital herpes simplex virus (HSV) infection (P = 10(-4)), an age inferior or equal to 30 years (P = 0.012), and cigarette smoking (P = 0.020). Crude and adjusted relative risks were calculated for each HPV persistence predictor. The data and the results of the molecular biology of high-risk genital HPVs are consistent with the hypothesis that the use of an oral contraceptive containing simultaneously and continuously both a potent estrogen and a high activity progestative is necessary to enhance significantly HPV transcription. These observations are also consistent with the hypothesis that the oral contraceptives and HSV genital infection are responsible for HPV persistence in the normal cervix but not for HPV-induced cervical transformation.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of rufloxacin in Salmonella typhimurium infection in mice. This study was undertaken to investigate the efficacy of rufloxacin, a new quinolone which is interesting due to its pharmacokinetics characterized by a long plasma half-life, in the treatment of systemic salmonella infections in the mouse typhoid model. Innately susceptible BALB/c and resistant CBA mice were used to investigate the efficacy of rufloxacin in controlling systemic salmonella infections when given for brief or prolonged periods. The present study shows that rufloxacin is not only very effective on both mouse strains, but can completely eradicate the salmonellae from livers and spleens when given early in the infection of CBA resistant mice.


Clinical comparative study of azithromycin versus erythromycin in the treatment of acute respiratory tract infections in children. The efficacy and tolerability of azithromycin and erythromycin in the treatment of acute respiratory tract infections in children were compared in an open, multicenter, randomized trial. A total of 151 children, aged from 2 months to 14 years, suffering from upper airways infections (60), or lower respiratory tract infections (91), were randomized to be treated either with azithromycin, 10 mg/Kg/day per os once daily for 3 or 10 mg/Kg/day 1 and 5 mg/Kg/days 2-5 (77 patients) or with erythromycin, 50 mg/Kg/day thrice daily for at least 7 days (74 patients). The two treatment groups did not significantly differ as to sex, age, weight, type and severity of infection, and infecting pathogens. Clinical evaluation was performed prior to therapy, on treatment days 1, 3, 5 and 7, and on day 10. Microbiological and laboratory assessment were carried out at baseline and after the end of therapeutic course. Chest X-ray and serologic assays for Mycoplasma pneumoniae infection were obtained in patients suspected to have lower respiratory tract infections. At the end of therapy, clinical cure was achieved in 73 out of 77 patients (94.8%) in the azithromycin group, and in 60/72 evaluable subjects (83.3%) in the erythromycin group. A significantly more rapid remission of several illness-related signs and symptoms was observed in patients treated with azithromycin. A total of 75 bacterial pathogens were isolated at baseline microbiological examination; at the end of the therapeutic course bacteriological eradication was obtained in 34/34 cases (100%) treated with azithromycin, and in 40/41 children (97.5%) treated with erythromycin.(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical use of the new macrolides, azalides, and streptogramins in pediatrics. Macrolides are the primary drugs of choice for a number of clinically significant infections in children. The clinical aspects of newer macrolides such as roxithromycin, clarithromycin, dirithromycin, flurithromycin, miocamycin, rokitamycin, azithromycin and RP 59500 are discussed in different pediatric infections including streptococcal infections (e.g. pharyngitis, otitis, pneumonia, skin infections), staphylococcus soft tissue infections, mycoplasma pneumonia, chlamydial infections as well as legionellosis and campylobacter enteritis. Also, incidences of adverse events in pediatric patients receiving different macrolides are indicated as well as the dosages in children. The advantages of newer macrolides are: lower dosages, b.i.d. or once daily dosage regimens, good intracellular and tissue penetration, better activity against gram-negative microorganisms (some) and a low rate of adverse reactions.


Antibacterial efficacy of root canal medicaments. The antibacterial efficacy of four endodontic medicaments was studied in vitro. Calcium hydroxide (CALACEPT), camphorated parachlorophenol (CPCP), Cresophene, and 2% iodine-potassium iodide (IKI) were tested against four anaerobic microorganisms, Streptococcus mutans, Peptostreptococcus anaerobius, Porphyromonus gingivalis and Fusobacterium nucleatum, for 10 or 15 min. IKI was effective only against F. nucleatum and P. gingivalis, and the others were effective against all four microorganisms.


Changes in the levels of rat interleukin 8/CINC and gelatinase in the exudate of carrageenin-induced inflammation in rats. A sensitive enzyme-linked immunosorbent assay (ELISA) for rat interleukin 8/cytokine-induced neutrophil chemoattractant (CINC) has been established by using biotin-conjugated anti-CINC rabbit IgG. The biotin-streptavidin sandwich ELISA detected CINC at concentrations from 3 pg/ml up to 30 ng/ml. The concentration of CINC in the pouch fluid (exudate) of rat carrageenin-induced inflammation was measured by the ELISA. After a time lag of about 2 h, neutrophils steadily accumulated in the carrageenin/air-pouch until 8 h. Similarly, the CINC level of exudate increased after about a 2-h lag, and reached a maximum (134 ng/ml) at 8 h, and thereafter decreased to a negligible concentration at 24 h after carrageenin injection. In association with the rise in CINC level, the concentration of exudate 96-kDa gelatinase corresponding to neutrophil gelatinase/type IV collagenase increased with time. The results suggest that CINC contributes, at least in part, to the neutrophil migration into the inflammatory lesion of the carrageenin-induced inflammation in rats.


Cyclic AMP mimics IL-1 action in augmenting the differentiation of a mouse myeloid leukemic cell line (M1). We have shown previously that recombinant human interleukin 1(IL-1) and interleukin 6 (IL-6) inhibited the proliferation of a mouse myeloid leukemic cell line (M1), and that IL-6 induced differentiation of the cells into macrophage-like cells and that IL-1 augmented this differentiation. Using this model we investigated the action mechanisms of IL-1 and IL-6. IL-6, but not IL-1, stimulated prostaglandin E2 (PGE2) production. The differentiative effect of IL-6 however, was not suppressed by indomethacin, although PGE2 induction by IL-6 was completely inhibited. Exogenously added PGE2 neither augmented the differentiative effect of IL-6 nor induced differentiation in combination with IL-1. Therefore, stimulation of PGE2 production did not appear to be essential for differentiative effects of these cytokines. Dibutyryl cAMP, 8-Br-cAMP and two adenylate cyclase-activating reagents, cholera toxin (CT) and forskolin (FK), all exhibited the similar augmenting effects as IL-1. These reagents augmented M1 cell differentiation by IL-6, and they did not induce differentiation in combination with IL-1. cAMP derivatives, CT, FK, IL-1 and IL-6 all inhibited the proliferation of M1 cells. CT and FK increased the intracellular cAMP levels. However, neither IL-1 nor IL-6 increased the cAMP levels. In contrast to the cAMP derivatives and reagents that activate adenylate cyclase activity, phorbol 12-myristate 13-acetate (PMA) and calcium ionophore neither induced nor augmented the differentiation in combination with either IL-1 or IL-6. Intracellular Ca2+ concentration was not altered by IL-1 or IL-6 suggesting that Ca2+/Calmodulin kinase and protein kinase C activation are not involved in this signal transduction pathway. Therefore, the present study suggests that IL-1 exhibits an effect similar to that of cAMP without affecting intracellular cAMP level.


[Chronic fatigue syndrome and virus infection: human herpesvirus 6 (HHV-6) infection]. Chronic fatigue syndrome (CFS) is newly-recognized disease characterized by chronic and debilitating fatigue. It has been suggested that viral infection may be involved in this syndrome from the results of clinical examination, including increased activity of 2',5'-synthetase in leukocytes of patients. The following viruses have been reported as etiologic agents of this disease. First, many studies have found elevated levels of IgG to viral capsid antigen and early antigens to Epstein-Barr virus (EBV), but low titer or absence of antibody to EBV-associated nuclear antigen. Second, the enteroviruses have also been implicated as possible causative agent of CFS, because virus could be isolated from patients. Recently it was also reported that antibodies to human T-lymphotropic virus (HTLV) and HTLV type II (HTLV-II) gag sequence were detectable in patients. Finally several reports state that human herpesvirus 6 (HHV-6) could be isolated from CFS patients in the high frequency. In conclusion, it is still early to identify the etiologic agent from these reports, and more effort is needed.


[Viral infection and its causative role for chronic fatigue syndrome]. Patients with chronic fatigue syndrome (CFS), of unknown etiology, have been increasingly reported. This syndrome is characterized by debilitating fatigue, lymphadenopathy, and fever. Herein, I focus on and review this syndrome from the view point of the causative role of viral infection. Since the symptoms of CFS are similar to those of chronic infectious mononucleosis (CIM) or chronic Epstein-Barr virus infection (CEBV), the role of EBV has been intensively studied. The etiological relationship between EBV and CFS, however, is questioned, like other lymphotropic viruses, including human retroviruses, adenoviruses and human herpesvirus 6. Additionally, severe chronic active EBV infection syndrome (SCAEBV) is also discussed in this review because symptoms of this disorder are similar to those of CFS but more severe in degree. Currently, the cause(s) and treatment of CFS are enigmatic and require further research and multidisciplinary study.


[Chronic fatigue syndrome--study of 51 cases treated at the Second Tokyo National Hospital]. Fifty-one patients with chronic fatigue syndrome (CFS) were studied. Tender points, which are a characteristic clinical feature of fibromyalgia, were found in all but two of the patients at 11.4 points (mean) per patient. IgG antibody titers to EB virus viral capsid antigen were more elevated in the CFS patient group compared to that of the control (p < 0.0015). IgG antibody titers to HHV-6 were not higher in the patient group. NK cell activity was not more decreased in the patient group, whereas, the mean number of NK cells was lower (p < 0.005) in the patient group, when CD57 was used as the NK cell marker. Viral infections and/or disorders in cellular immunity may be important factors in the pathogenesis of CFS.


[Chronic fatigue syndrome--cases in the Kanebo Memorial Hospital]. In our hospital, 134 patients (28 male, 106 female, 10-82 years of age) were diagnosed as having chronic fatigue syndrome (CFS). Some patients had mild elevation of antibodies against Epstein-Barr Virus and immunologic abnormalities (natural killer cell dysfunction and high rates of skin reactivity to house dust, pollen, drugs and common food). In the patients with immunologic abnormalities, we found decreases in serum concentrations of arachidonic acid and dihomogamma-linolenic acid. A Kampo medicine, Ren-Shen-Yang-Rong-Tang was used in the management of 134 patients and 98 patients returned to work or school.


[Symptoms, signs and laboratory findings in patients with chronic fatigue syndrome]. This review summarizes the symptoms, signs and laboratory abnormalities seen in 59 patients with chronic fatigue syndrome (CFS), 2 patients with post-infectious CFS and in 26 patients with possible CFS whose illnesses fulfill the criteria proposed by the study group of the Ministry of Welfare, Japan. The characteristic symptoms and signs of CFS are prolonged generalized fatigue following exercise, headache, neuropsychological symptoms, sleep disturbance and mild fever. In possible CFS patients, the frequency of mild fever, muscle weakness, myalgia and headache is low. Our standard hematologic and laboratory tests revealed a few abnormality in patients with CFS. The characteristic abnormality in CFS patients is the low values of 17-Ketosteroid-Sulfates/creatinine in morning urine and the acylcarnitine deficiency. It seems likely that this deficiency of acylcarnitine induces an energy deficit in the skeletal muscle, resulting in general fatigue, myalgia, muscle weakness and postexertional malaise in CFS patients. Virologic studies revealed no evidence of retrovirus infection with HTLV-1, HTLV-2 and HIV, but the reactivation of HHV-6 infection was apparent.


[Chronic fatigue syndrome, a case of high anti-HHV-6 antibody titer and one associated with primary hyperaldosteronism]. Two cases of chronic fatigue syndrome (CFS) were reported which were suggestive for the study of the etiology and a cure for CFS. Case 1: A 31-year-old woman was admitted for chronic fatigue syndrome. Examination revealed a high titer of anti HHV-6 antigen of x2560 and an increased percentage of suppressor T lymphocytes in the peripheral blood. HHV-6 was speculated to be reactivated and stimulating the immune system in CFS. Case 2: A 46-year-old woman suffering from CFS had been in remission for 6 years. She was admitted for hypertension associated with right adrenal adenoma and hyperaldosteronism. After right adrenalectomy, there was a recurrence of high fever and other CFS symptoms. It was suggested that CFS symptoms may be ameliorated by aldosterone.


[Brain-gut interactions in irritable bowel syndrome: physiological and psychological aspect]. Recent advances in the investigation of brain-gut interaction in irritable bowel syndrome (IBS) were reviewed. Brain is suggested to play an important role in the pathophysiology of IBS on the basis of the following evidence. (1) Stress often induces major symptoms of IBS patients (Drossman et al., 1982), simultaneously with colonic hypermotility (Fukudo et al., 1987) or dysmotility of the small intestine (Kumar et al., 1985). (2) IBS patients rarely express symptoms or small intestinal dysmotility during sleep (Kellow et al., 1990). (3) IBS patients complain of more pain with balloon distension of the colon or rectum than normal controls; visceral perception is enhanced in IBS (Whitehead et al., 1990). (4) IBS patients often show psychoneurotic symptoms and extra-colonic somatic symptoms (Young et al., 1976). (5) There are some animal (Williams et al, 1987) or human (Dinan et al, 1990) experiments which indicate the possible involvement of brain peptide or brain monoamine in IBS. (6) Dysrhythmia or increased beta power in electroencephalogram is observed more often in IBS patients than in the normal controls (Fukudo et al, 1991) in addition to abnormal REM sleep in IBS patients (Kumar et al., 1992). These observations support our hypothesis that not only the gut but also the brain show dysfunction and exaggerated responsivity to the stimuli in IBS. Further research on brain-gut interaction in IBS is warranted.


[Dietary treatment in patients with irritable bowel syndrome]. The author claims that dietary treatment of irritable bowel syndrome (IBS) consists of methods aiming at improvement of abdominal symptoms and functional disorder of the bowel. Patients with constipation are recommended to take dietary fiber positively, while those with diarrhea should consume sparingly food which may cool their body. Both should avoid overeating and overdrinking, and have regular dietary habits. In order to improve the functional disorder of the bowel, it is necessary for those patients (1) to be careful not to take often refined cereals or manufactured foods, (2) to eat green and yellow vegetables and seaweeds positively, as well as, protein and fat in proper quantity, and (3) to take care of the well-balanced intake of various kinds of vitamins, minerals and other nutriments.


Involvement of alpha 2-adrenoceptors in the sacral micturition reflex in rats. We have studied the effects of intrathecally-injected drugs that act on alpha-adrenoceptors in the urinary bladder reflex contractile activity evoked by continuous infusion of fluid into the bladder of anesthetized rats. Clonidine (10 and 30 micrograms) facilitated and yohimbine (100 micrograms) abolished the bladder contractile activity, and pretreatment with yohimbine (30 micrograms) inhibited the effect of clonidine (10 micrograms). Phenylephrine (60 micrograms) abolished the bladder contractile activity, but prazosin (40 micrograms) had no significant effect on it. The bladder contractions induced by electrical stimulation of the pontine micturition center were inhibited by yohimbine in a dose-dependent manner. These results suggest that transmission in the descending neurons from the pontine micturition center to the sacral parasympathetic neurons that control bladder motility is mediated by alpha 2-adrenoceptors in rats.


[Neoplastic space-occupying lesions of the orbits. II. Space-occupying lesion in the area of the lacrimal gland]. In part two of this paper about orbital tumors, neoplasms of the lacrimal gland are discussed: These have to be properly separated from inflammations. While inflammatory affections of the lacrimal gland make up the majority of private practice consultations, in cases referred to an eye clinic the relation between inflammatory diseases and neoplasms is about equal (between 1987 and 1990, 14 neoplasias and 14 inflammations were seen at the University Eye Clinic, Zurich). The benign pleomorphic adenoma of the lacrimal gland should be removed in toto in its capsule. For this procedure a lateral orbital fenestration is required. An excisional biopsy is considered the method of choice while an incisional biopsy should be avoided because of the risk of recurrence. On the other hand, incisional biopsy is used in the cases of adenoidcystic carcinoma and lymphoma. After confirmation of an adenoidcystic carcinoma by biopsy, orbital exenteration has to follow as soon as possible. In cases of lymphoma, possible oncological treatment has to be evaluated. In order to differentiate histologically between a reactive lymphoid hyperplasia and a malignant lymphoma, immunofluorescent studies on non-fixed tissue are necessary in close collaboration with an immuno-pathologist. Because the benign pleomorphic adenoma of the lacrimal gland has to be treated by a different surgical approach than the adenoid cystic carcinoma, a proper diagnosis has to be made before any intervention; a requirement we could not always fulfill. Those mistakes made us conclude that even in the era of CT scan, MRI and angiography, the radiological diagnoses are often hypotheses which have to be confirmed by discussion.(ABSTRACT TRUNCATED AT 250 WORDS)


A simple biochemical approach to differentiate synaptosomes and non-synaptic mitochondria from rat brain. A simple and rapid low speed density gradient centrifugation method has been described to isolate relatively pure synaptosomes and non-synaptic mitochondria from rat brain cerebral cortex. The purity of the fractions isolated were tested biochemically by considering some marker enzymes, i.e., lactate dehydrogenase, plasma membrane Na+K+ATPase and acetylcholinesterase for the synaptosomes and succinate dehydrogenase for the mitochondrial fraction. The adrenergic receptor properties in the synaptosomal membrane were evaluated by observing the effect of different beta- and alpha 2-adrenergic receptor agonists and antagonists as function of synaptic plasma membrane Na+K+ATPase activity. Isoproterenol (beta-agonist) and yohimbine (alpha 2-antagonist)-induced changes in the synaptosomal Na+K+ATPase activity were counteracted by beta-antagonist propranolol and alpha 2-agonist clonidine, respectively. In the non-synaptic mitochondria the corresponding effects were insignificant. The study illustrates an easy, rapid and low-speed preparatory method to obtain synaptosomal and non-synaptic mitochondrial fractions of high purity.


Isolation of herpes simplex virus-1 by intracellular membranes of BHK tk- cells. Baby hamster kidney (BHK tk-) cells infected with herpes simplex virus-1 (HSV-1) showed a large number of virus particles isolated in vesicles characterized by the presence or the absence of ribosomes or inside cisternae of the rough endoplasmic reticulum or the nuclear envelope. The isolation of the virions by intracellular membranes appeared shortly after infection of the cells by HSV-1. These structures persisted for longer periods where no morphological alterations in the infected cells were noted as well as at periods where expression of the late viral genes and the presence of empty capsids or DNA-containing new capsids in the nucleoplasm of BHK tk- cells were detected. The results suggest that the presence of virions in membranic formations of the infected cells may be an indication of permanent isolation and subsequent deactivation of the viruses rather than an intermediate stage during their transport from the plasma membrane to the nucleus. The possible mechanisms by which the virions are isolated by the intracellular membranes of BHK tk- cells are discussed.


Enteric adenovirus type 41 isolates: cloning, physical maps and diversity in restriction enzyme cleavage pattern. Adenovirus (Ad) type 40 and 41 DNAs were directly extracted from stool specimens of children with gastroenteritis. Two new strains of Ad41, Sanekata and Ehime strain, were cloned and their restriction maps were constructed. The left terminal end of the cloned Ad41 genome, EcoRI-E fragment of the Sanekata strain and EcoRI-F fragment of the Ehime strain, had transforming ability in rat 3Y1 cells. Only one of the 35 isolates of Ad40 tested showed a different restriction profile, while three different restriction profiles were found in DNAs from Ad41 isolates.


Augmentation of bovine leukemia virus (BLV)-specific lymphocyte proliferation responses in ruminants by inoculation with BLV env-recombinant vaccinia virus: their role in the suppression of BLV replication. Lymphocyte proliferation responses were investigated in sheep and cattle, in which the replication of bovine leukemia virus (BLV) had been known to be suppressed by inoculation with recombinant vaccinia virus (rVV) expressing BLV envelope glycoprotein (gp60). Enhanced lymphocyte proliferation responses were observed in animals inoculated with rVV, regardless of whether they were naive or BLV carriers. These responses were roughly inversely correlated to the growth of BLV in the peripheral blood leukocytes. In contrast, there was no apparent correlation between humoral immune response and BLV growth. Based on these results, it was suggested that rVV rendered its suppressive effect of BLV replication primarily via augmentation of cell-mediated immunity.


Replication of human cytomegalovirus in the cells of the U937 monocytoid cell line. Human cytomegalovirus (HCMV) infection in immunocompromized hosts sometimes occurs as a result of reactivation. Cells of the monocyte-macrophage linkage are suggested to be a site of latency and persistence for HCMV. The human monocytic cell line U937 was infected with the AD169 strain and a clinical isolate of HCMV. The expression of surface antigens on the cells was assessed by flow cytometry. The polymerase chain reaction (PCR) was used to detect viral DNA from infected cells. CMV immediate early antigen, early antigen, and late antigen (LA) were detected from both clinical isolate- and AD169-inoculated U937 cells by flow cytometry. CMV DNA which code major immediate early gene (US3) and LA gene (US14) were detected from the clinical isolate-inoculated U937 over a period of 31 days as tested by PCR. These U937 cells proliferated as well as uninfected U937 cell, but only a small number of AD169-inoculated U937 cells survived after 14 days of inoculation. Interleukin-2 activities were detected in the media on days 24-40 after inoculation with AD169. This chronic CMV infection model of U937 might be utilized to study the mechanisms of persistence and reactivation.


[Immunohistochemical studies on tumor associated carbohydrate antigens in gastric cancers of different histological stages]. By comparing gastric cancer tissues treated by the conventional hematoxylin and eosin (HE) staining and those by the chemical staining of immunohistologicals using monoclonal antibodies (MoAB) which recognize different carbohydrate antigens, the relation of cancer tissue patterns between the two staining methods was studied. Consequently, stainability was not seen in MoAB-FH4, AH6, FH6 and TKH2 in the cancer tissues where MoAB-SH1 responded to immunohistological staining. Likewise, each of MoAB-FH4, AH6, FH6 was found to have its own stain localization. The patterns made by immunohistological staining using MoAB showed so-called mosaicism even where the HE stain presented the same histologic form. Study of correlation between gastric cancer patterns and MoAB's localization revealed that localization of MoAB-SH1, AH6 and TKH2 was predominant in well differentiated adenocarcinoma. On the contrary, MoAB-FH4 and FH6, which are more specific, showed predominant localization in poorly differentiated adenocarcinoma of gastric cancers. Localization of MoAB-FH6 and AH6 increased as cancer grew from the early stage to the advanced stage. These results leads to this assumption: cancer, being of an isogenic carbohydrate structure at the initial stage when carcinoma in situ is generated, gains heterogeneity with the process of growth, differentiating into various directions and thus changing into a complicated carbohydrate structure.


Retinoic acid receptor isoform RAR gamma 1: an antagonist of the transactivation of the RAR beta RARE in epithelial cell lines and normal human keratinocytes. The diversity of isoforms of retinoic acid (RA) receptors (RARs) and of DNA sequences of retinoic acid-responsive elements (RAREs) suggests the existence of selectivities in the RAR/RARE recognition or in the subsequent gene modulation. Such selectivities might be particularly important for RAREs involved in positive feedback, eg. the RAR beta RARE. In the present work we found that in several epithelial cell lines, reporter constructs containing the RAR beta RARE linked to the HSV-tk promoter were transactivated in the presence of RA by endogenous RARs and co-transfected RAR alpha 1 and RAR beta 2 isoforms, but not by RAR gamam 1. On the contrary, this latter isoform behaved towards the RAR beta RARE as an inhibitor of the transactivation produced by endogenous RARs and by cotransfected RAR alpha 1 and RAR beta 2. RAR gamma 1 also behaved as an antagonist of the transactivation produced by cotransfected RXR alpha. The natural RAR beta gene promoter or RAR beta RARE tk constructs were not activated by the endogenous receptors of normal human keratinocytes (NHK), which are known to contain predominantly RAR gamma 1. It was, however, possible to activate to a certain extent RAR beta RARE-reporter constructs in NHK by co-transfecting RAR alpha 1, RAR beta 2 or RXR alpha. The antagonist behavior of RAR gamma 1 towards the RAR beta RARE may explain why in certain cell types such as keratinocytes, RAR beta is neither expressed nor induced by RA.


Shuttle vectors conferring hygromycin B resistance to E. coli and to mammalian cells. Differential expression of carboxyterminal fusion proteins. Shuttle vectors expressing resistance to hygromycin B in both E. coli and in mammalian cells were constructed. A combination of the simian virus 40 early promoter upstream of the native bacterial promoter of the neo gene from transposon Tn5 was found to express hygromycin B resistance better in both types of host cells than a combination of the Tn5 promoter followed by the promoter of the Herpes simplex virus thymidine kinase gene. Hygromycin phosphotransferase fusion proteins with extensions at the carboxyterminus were also tested and found to be marginally less effective as selection markers in eukaryotic cells but virtually inactive in E. coli.