A Reovirus Causes Hypovirulence of Rosellinia necatrix.

ABSTRACT White root rot, caused by Rosellinia necatrix, is a serious soilborne disease of fruit trees and other woody plants. R. necatrix isolate W370 contains 12 segments of double-stranded RNA (dsRNA) that is believed to represent a possible member of the family Reoviridae. W370 was weakly virulent and its hyphal-tip strains became dsRNA free and strongly virulent. The 12 segments of W370dsRNA were transmitted to hygromycin B-resistant strain RT37-1, derived from a dsRNA-free strain of W370 in all or none fashion through hyphal contact with W370. The W370dsRNA-transmitted strains were less virulent than their parent strain RT37-1 on apple seedlings, with mortality ranging between 0 to 16.7% in apple seedlings that were inoculated with the W370dsRNA-containing strains and 50 to 100% for seedlings inoculated with the dsRNA-free strains. Some W370dsRNA-containing strains killed greater than 16.7% of seedlings, but these were found to have lost the dsRNA in planta. These results indicate that W370dsRNA is a hypovirulence factor in R. necatrix. In addition, a strain lost one segment (S8) of W370dsRNA during subculture, and the S8-deficient mutant strain also exhibits hypovirulence in R. necatrix.

Acetyl-low density lipoprotein receptors on rat mesangial cells.

To elucidate whether mesangial cells have any scavenger functions for modified lipoproteins, surface binding and cholesteryl ester (CE) formation by acetyl-low density lipoproteins (acetyl-LDL) have been studied in cultured rat renal mesangial cells. Specific binding kinetics for acetyl-LDL were observed with Kd = 28.3 micrograms/ml and Bmax = 1.1 ng/micrograms cell protein at 0 degrees C. The fluorescence microscopic finding demonstrated the enhanced uptake of DiI-acetyl-LDL in mesangial cells. Incorporation of [14C]oleate into CE was enhanced to 6-fold by loading 30 micrograms/ml of acetyl-LDL on 10 micrograms/ml of [14C]oleate-bovine serum albumin conjugate as compared with the control without lipoproteins (P < 0.05). The CE formation was completely inhibited by chloroquine. The light microscopic finding demonstrated the increased CE deposition by acetyl-LDL, resulting in foam cell formation. These results indicate biochemically and morphologically that the mesangial cells take up acetyl-LDL by receptor-mediated endocytosis, and that cholesterols in acetyl-LDL are converted to CE, resulting in an increased cellular cholesterol content. In conclusion, mesangial cells may have a scavenger function similar to macrophages.

Phenol block for hip flexor muscle spasticity under ultrasonic monitoring.

Hip flexor spasticity, which is often associated with central nervous system (CNS) diseases, is a major impediment in rehabilitation. In order to cope with this problem, lumbar nerve blocking techniques developed by Meelhuysen and major and minor psoas muscle blocking techniques developed by Awad have been used in combination with physical therapies. Based on these techniques, we conducted major and minor psoas muscle phenol block (motor point block or intramuscular nerve block) under ultrasonic monitoring. Phenol block was conducted in nine patients with cerebral infarction (13 blocking procedures) and three with spinal cord injuries (six blocking procedures) while keeping them in a lateral position with the operation side upside. The beginning of the femoral nerves and part of the lumbar artery were visualized by ultrasound in some patients. As a result of the improvement of hip flexor spasticity, the range of hip joint motion (determined by the Mundale technique, prone hip extension and Thomas test) improved shortly after blocking. When physical therapy was conducted after blocking, improvement of skin care management was observed in eight cases, ability to keep in a stable sitting position in nine, improvement of a standing posture in three, increases in the ability to walk in two and alleviation of pain in three. Although nerve block is reported to result in hematoma, decreases in muscle force, pain, cystic/rectal disorders and hypogonadism, we have observed no such complication in our patients.

A specific binding site for lipoproteins containing apolipoprotein A-I on human hepatoma cell line HepG2.

Interactions of lipoproteins containing apolipoprotein (apo) A-I with or without apoA-II with human hepatoma cell line HepG2 were studied to investigate the ligand specificity for high density lipoprotein receptor on human hepatic cells and their metabolism. The two types of lipoproteins were isolated by immunoaffinity chromatography, in which apoE-containing lipoproteins were removed. Specific binding kinetics at 0 degrees C were observed for the apoA-I-containing lipoproteins with or without apoA-II (Kd = 18 or 20 micrograms protein/ml, Bmax = 110 or 120 ng/mg cell protein, respectively). The binding of these lipoproteins to HepG2 cells was competitively inhibited by excess unlabeled apoA-I-containing lipoproteins or apoA-I-phospholipid complexes, but not by apoA-II.phospholipid complexes. Interactions of these lipoproteins with HepG2 cells at 37 degrees C were further examined. These results suggested that HepG2 cells have a specific binding site for apoA-I-containing lipoproteins, and that apoA-I might be a crucial component in the binding of these lipoproteins to human hepatic cells.

Placetins, platelet aggregation inhibitors from Streptomyces sp. Q-1043. I. Fermentation, isolation and biological properties.

Placetins, platelet aggregation inhibitors were obtained from the culture broth of Streptomyces sp. Q-1043. These were designated placetins A, A1, B and B1, respectively. Placetins A and B showed strong cytotoxicities against P388, L1210 and HeLa cells.

[An evaluation of MR imaging of soft-tissue masses in extremities].

We evaluated 22 soft-tissue masses in the extremities using magnetic resonance (MR) imaging. The twenty-two cases consist of 14 benign lesions and 8 malignant lesions. MR imaging is available to provide the representation of the extent of the masses, though characteristic diagnosis of them could not be identified. We think that there are more advantages to using MR imaging rather than computed tomography (CT) in the evaluation of soft-tissue masses in extremities.

Effects of plasmapheresis on familial type III hyperlipoproteinemia associated with glomerular lipidosis, nephrotic syndrome and diabetes mellitus.

A 59-year-old woman, one of 5 cases with familial type III hyperlipoproteinemia reported at our clinic to date, had nephrotic syndrome and diabetes mellitus, but had neither coronary atherosclerosis nor xanthoma. A renal biopsy specimen revealed a massive cluster of foam cells containing apolipoprotein B and E in the mesangial region of the kidney. A restricted diet intake combined with lipid-lowering drugs such as cholestyramine, clinofibrate, and bezafibrate, in addition to methylprednisolone was not very effective in lowering serum triglyceride and cholesterol levels within physiological ranges. Therefore, plasmapheresis, using a dextran sulfate-cellulose column, was performed. Repeated plasmapheresis resulted in a marked decrease in both serum total cholesterol and triglyceride. A second renal biopsy specimen performed 2 years later revealed a marked reduction in foam cells with concurrent improvement in her nephrotic syndrome and glucose intolerance. These results suggest that familial type III hyperlipoproteinemia may be responsible for glomerular lipidosis resulting in nephrotic syndrome. They also indicate that plasmapheresis using a dextran sulfate-cellulose column is very effective in the removal of abnormal lipoproteins such as beta-very low density lipoprotein and intermediate density lipoprotein in a case of familial type III hyperlipoproteinemia.

[MRI imaging of liver neoplasma].

MRI by 1.5 T imager with spin echo pulse sequence was performed on 53 liver neoplasms (13 hepatocellular carcinomas (HCC), 28 cavernous hemangiomas and 12 metastasis). Lobulated and notched contour were observed in 72% of our cases of hemangioma. It is well known that in hemangioma several kinds of echographic patterns appear, but in MRI, almost all hemangioma appear to show the same signal pattern as high intensity homogeneous on T2 weighted image. No liver neoplasma showed high intensity on T1 image except in 1/3 of cases of HCC. Contrast enhancement with Gd-DTPA is useful for differential diagnosis of liver neoplasma because it shows different signal patterns from the other diseases.

Effect of alpha 2-adrenoceptor antagonist on platelet activation during insulin-induced hypoglycaemia in type 2 (non-insulin-dependent) diabetes mellitus.

The role of epinephrine in platelet activation and the effect of an alpha 2-adrenoceptor antagonist, midaglizole, during insulin-induced hypoglycaemia in Type 2 (non-insulin-dependent) diabetes mellitus were examined. The action of midaglizole as a platelet alpha 2-antagonist was confirmed by in vitro studies using platelet-rich plasma and washed platelet suspension. Hypoglycaemia was induced by a bolus injection of short-acting insulin in 24 diabetic patients. They were divided into two groups, a control group (n = 12) and an alpha 2-group (n = 12), and midaglizole was administered orally 60 min before insulin injection in the latter. Blood glucose and plasma C-peptide levels were significantly decreased (p less than 0.005) by insulin injection in both groups. Counter-regulatory hormones, including epinephrine, and arginine vasopressin were similarly increased at the hypoglycaemic nadir compared with the levels at 0 min in both groups. Plasma beta-thromboglobulin was increased at the hypoglycaemic nadir (165.5 +/- 12.6 ng/ml) compared with the level at 0 min (121.0 +/- 11.5, p less than 0.005) in the control group, whereas no significant increase was demonstrated in the alpha 2-group. These results suggest that plasma epinephrine plays an important role in platelet activation during hypoglycaemia in Type 2 diabetes mellitus, and that the platelet activation is prevented by alpha 2-adrenoceptor antagonist.

Chronic granulomatous disease with neutrophil membrane cytochrome b deficiency: demonstration by immunochemical staining with monoclonal antibody.

Cytochrome b deficiency in the peripheral granulocytes of two male patients with chronic granulomatous disease was demonstrated by an immunocytochemical assay using a monoclonal antibody, 7D5, against human neutrophil cytochrome b. A mosaic of cytochrome b positive and negative neutrophils, indicating a carrier state in an X-linked trait, was found in the mother of patient 1 but not in the mother of patient 2.

Idiopathic thrombocytopenic purpura subsequent to Graves' disease and insulin-dependent diabetes mellitus.

We report a woman with idiopathic thrombocytopenic purpura (ITP) subsequent to Graves' disease and insulin-dependent diabetes mellitus (IDDM). The patient was affected by Graves' disease at the age of 29 and developed IDDM the following year. The ITP occurred after 8 years of well-controlled Graves' disease and IDDM with appropriate treatment, but subsided with prednisolone therapy, followed by splenectomy. Graves' disease was observed also in the parents. The serum anti-thyroid-microsome antibody, anti-parietal-cell antibody, anti-pancreas-islet-cell antibody and anti-platelet antibody were all positive. HLA analysis revealed that haplotypes including DR4, DRw9 and DRw53 were positive in both the patient and her mother. In Japanese IDDM patients with autoimmune diseases, these three haplotypes have been reported to be significantly frequent. These findings indicate that the patient of this study is a typical case of the primary autoimmune type of diabetes mellitus proposed by Irvine and Bottazzo. However, the combination of Graves' disease, IDDM and ITP has not been hitherto reported to our knowledge.

[A new technique for proximal esophagography and quantitative analysis of pharyngeal function].

The roentgenologic examination of the upper esophagus in somewhat difficult due to the rapid passage of barium. Therefore we designed a new technique for proximal esophagography, using magnetic video disc recorder and multiformat camera (VDR-MC technique). The video signal from fluoroscopic unit is led to video disc recorder (VDR) and after the examination have been finished the recorded images are transferred into multiformat camera, and then the images are copied sequentially on a sheet film VDR have a storage capacity, 600 frames per 20 seconds. The sequential pictures of barium swallowing in one deglutition movement are shown as the time-intervals of each 0.03 seconds on a sheet film. The movement of barium bolus in one deglutition will be demonstrated exactly and except any failure. This working process of VDR-MC technique in very simple and convenient, and when the reading and diagnosis of X-ray appearances, we can easily compare with the changes of each image. This is a characteristic merit of new technique. In order to examine the function of deglutition we designed a new technique by means of the dynamic pharyngo-esophagrams as before. The recorded dynamic images are transferred into multiformat camera and copied on a sheet film by the intervals of 0.03 seconds in turn. Viewing these sequential images the each measuring points are determined ane the barium transit times are calculated from the number of frames, and then transferred to the time in millisecond unit. According to the results of experiment, the measured values well correspond to that by EMG on the pharyngeal movement of deglutition. These quantitative analysis on the pharyngeal function and morphological diagnosis will be performed in easy procedure at the same time. The outline of the discussion about the advantages of this technique is as follows; The performance of the examination by new technique will be done with extreme easy. A difficult examination will be performed without failure and finished in a short time successfully. We can examine enough the patients who are not able to understand the doctor's indication, for example children and patients having hard hearing. In comparison with cinefluorographic unit, development of films, patient's position, observation and copy of images and storage of film, became extremely easy. A quantitative measurement on pharyngeal function was designed and the reproducibility of measurements was given proof.

CCXVII. Separation and characterization of bile acid 3-glucuronides in human urine by high-performance liquid chromatography.

The separation and characterization of unconjugated and conjugated bile acid 3-glucuronides in biological fluids without prior deconjugation by high-performance liquid chromatography (HPLC) are described. A urine sample from a patient with obstructive jaundice was passed through a Sep-Pak C18 cartridge and was separated into groups by ion-exchange chromatography on a lipophilic gel, piperidinohydroxypropyl Sephadex LH-20, providing the glucuronide fraction. Subsequent resolution into individual 3-glucuronides was attained by HPLC on muBondapak C18 and Shodex ODS Pak F-411 columns. The 3-glucuronides of cholate, deoxycholate, chenodeoxycholate, glycocholate, glycochenodeoxycholate and taurochenodeoxycholate were identified on the basis of their behaviour in HPLC using mobile phases of different pH. The enzymatic hydrolysis of these glucuronides and derivatization of deconjugated bile acids with 1-anthroyl nitrile followed by chromatographic separation on a Cosmosil 5C18 column with fluorescence detection were carried out for unequivocal characterization. The ratio of unconjugated, glyco- and tauro-conjugated bile acid 3-glucuronides excreted in urine was found to be ca. 2:3:1.

Studies on steroids. CCXVI. Separation of bile acid 3-glucuronides by high-performance liquid chromatography.

The separation of 3-glucuronides of cholate, chenodeoxycholate, deoxycholate, ursodeoxycholate and lithocholate, and their glyco- and tauro-conjugates, has been carried out by high-performance liquid chromatography on a reversed-phase column. The chromatographic behaviour of bile acid 3-glucuronides was dependent on the type of conjugation. An effect of the pH of the mobile phase on the capacity ratio was observed at higher pH for chenodeoxycholate 3-glucuronide, probably owing to steric interaction of the 7 alpha-hydroxy group with the carboxy group in the glucuronyl moiety. Conversion of the alpha-hydroxy function on the steroid nucleus into an oxo group resulted in a 50% decrease in the capacity ratio. Bile acid 3-glucuronides were efficiently separated on Shodex ODS Pak F-411 using three kinds of ammonium phosphate buffer-acetonitrile systems.

Effect of simultaneous infusion of a somatostatin analogue, insulin and glucose on serum triglycerides and blood glucose levels in man.

Nine non-diabetic, non-obese, normocholesterolemic normal male subjects with varied triglycerides levels were subjected to a simultaneous infusion test with a synthetic somatostatin analogue [des(Ala1, Gly2)-D-Trp8, D-Asn3, 14-somatostatin], insulin and glucose under ambulatory conditions. The levels of C-peptide reactivity, immunoreactive glucagon and growth hormone were reduced, and the level of immunoreactive insulin remained constant during the infusion. The blood glucose reached a constant value at 110-120 minutes (steady state blood glucose, SSBG) after the commencement of the infusion. The total cholesterol (TC) levels decreased slightly in the 30 minutes after the experiments were begun, and the triglycerides (TG) levels decreased gradually throughout the infusion period, due mainly to the reduction of very low density lipoprotein (VLDL). The most striking finding was the highly significant positive correlation (p less than 0.005, r = 0.868) between SSBG and the serum TG level prior to the infusion. These results indicate an important relationship between insulin sensitivity and serum TG level. High TG level may be regarded as one of the indices of insulin resistance.