The effect of passages during Japanese BCG vaccine production on genetic stability and protective efficacy

Many genetic differences have been found among currently available BCG vaccines. To avoid continued accumulation of phenotypic or genotypic changes in the strains, WHO and most national regulatory authorities request that the vaccine should not be prepared by more than 12 passages from the master seed lot. However, it has recently been reported that genetic changes occur even during the passage for vaccine production. In this study, the genetic stability of Japanese BCG vaccine production using currently available PCR methods and protective efficacy using a guinea-pig model during the passages were examined. The results showed that there were no significant differences between the seed lot, the product manufactured by normal procedures, and the 20th passage product. These results indicate that the maximum number of passages as currently required by WHO for BCG vaccine production is adequate for the Japanese vaccine, and that new genetic tools may help to examine the quality control of the BCG vaccine.     

Progressive systemic sclerosis associated with primary small cell carcinoma of the stomach

A 64-year-old man with a 5-year history of progressive systemic sclerosis (PSS) was hospitalized because of melena. Radiological and endoscopic examinations showed an ulcerative lesion with sharply demarcated and raised margins in the fornix of the stomach. Tumor markers—serum carcinoembryonic antigen (CEA, 11.3 ng/ml) and neuron-specific enolase (NSE, 38.9 ng/ml) were elevated. Histological examination of endoscopic biopsy specimens (and of necropsy specimens) showed proliferation of atypical small round cells. Immunohistological examination of these cells showed they were positive for epithelial membranous antigen (EMA), and neuron-specific enolase (NSE), but negative for UCHL1, leukocyte common antigen (LCA), anti-leukocyte B-cell (MB1), and antileukocyte T-cell (MT1) antigens. Based on these histological and immunohistological tests, a definite diagnosis of small cell carcinoma of the stomach with PSS was established. Our case is a rare combination of PSS and gastric small cell carcinoma. We also reviewed the literature for the association between PSS and gastric cancer in Japanese patients.     

Scrotal Ulcers Arising during Treatment with All-trans Retinoic Acid for Acute Promyelocytic Leukemia

All-trans retinoic acid (ATRA) is effective in approximately 90% of the cases of acute promyelocytic leukemia (APL) with a low incidence of adverse effects. We report a patient with APL who developed skin ulcers of the scrotum concomitant with high fever during treatment that included ATRA. Severe fever was promptly alleviated with discontinuation of ATRA, while the ulcers improved gradually over 3 months. As the clinical features are similar to those of Sweet's syndrome, we should be aware of the possibility that this rare adverse effect may occur in the treatment with ATRA.     

The arterial ketone body ratio and serum α-fetoprotein level in patients with acute hepatic failure

Abstract: Hepatocyte regeneration is essential for recovery in acute hepatic failure, although it requires a large amount of energy. The ratio of acetoacetate to beta-hydroxybutyrate in arterial blood has been reported to reflect the cellular energy charge of hepatocytes, and we proposed that the recovery of the ratio in the early days of acute hepatic failure is essential for survival. However, there is no report on any marker of regeneration to confirm this hypothesis. In this study, we have assessed this ratio and the serum α-fetoprotein level sequentially in 26 patients with acute hepatic failure. Ten patients recovered and 16 died. The arterial blood ketone body ratio 3 days after the onset of hepatic encephalopathy of grade II or more was below 0.6 in 15 of the 16 nonsurvivors, whereas that in the 10 survivors was above 0.6. There was a positive correlation between the arterial blood ketone body ratio and the maximal concentration of α-fetoprotein (r=0.465, p<0.02 by Student's t-test). These data indicate that the arterial blood ketone body ratio is a marker for the capacity of the liver to regenerate and for the prognosis in patients with acute hepatic failure: effective hepatocyte regeneration may be impossible if these metabolic changes in acute hepatic failure impair the hepatocyte energy charge severely.     

Conformation of cyclo(-L-Pro-Gly-)(3) and its Ca and Mg complexes

The synthetic hexapeptide cyclo(-L-Pro-Gly-)₃ is an ionophore that shows interesting conformational changes upon binding metal ions. X-ray crystallographic studies of this peptide show that when it is crystallized from an ethanol/ethyl acetate mixture the ring takes up an asymmetric conformation containing one cis peptide bond. In crystals of a Ca²⁺ complex, the cation is sandwiched between two peptide molecules that differ markedly in conformation. However, both exhibit threefold symmetric forms, with all six peptide bonds in the molecule occurring in the usual trans conformation. The Ca²⁺ is octahedrally surrounded by six glycyl carbonyl oxygens from the two peptides at an average distance of 2.26 Å and can easily be released by the disruption of the peptide sandwich. In the magnesium complex, the peptide forms a 1:1 complex with the ion. The Mg²⁺ is octahedrally coordinated to three glycyl carbonyls and three water oxygens. The average coordination distance between magnesium and the peptide oxygens is 2.03 Å and that between magnesium and water oxygen is 2.11 Å. The two peptide molecules in the asymmetric unit have similar conformations and have approximate threefold symmetry.     

Endoscopic studies on the minute structures of colonic mucosa in the follow-up observation of ulcerative colitis

Journal of Gastroenterology - Thirty cases of ulcerative colitis were examined endoscopically by means of magnifying colonoscope with dye spraying method (the combined method), and their minute...     

Sulfur-containing amino acids attenuate the development of liver fibrosis in rats through down-regulation of stellate cell activation

BACKGROUND/AIMS: We tested the pharmacological action of sulfur-containing amino acids on the development of liver fibrosis in rats and on the function of cultured stellate cells. METHODS: Liver fibrosis was induced in rats by thioacetamide administration or by ligating the common bile duct. DNA synthesis of cultured stellate cells was evaluated by BrdU incorporation. The expression of proteins and phospho-proteins was determined by western blot analysis. mRNA expression was evaluated by RT-PCR. RESULTS: Oral administration of l-cysteine or l-methionine attenuated the deposition of collagen in liver tissues in the two fibrotic models, accompanying a reduction in the expression of smooth muscle alpha-actin and platelet-derived growth factor receptor beta and mRNAs of collagens, transforming growth factor-betas and tissue inhibitors of matrix metalloproteinase. In cultured stellate cells, l-cysteine and l-methionine suppressed the DNA synthesis and the expression of growth factor receptors, smooth muscle alpha-actin and type I collagen. They hampered the phosphorylation of p44/42 MAPK and Akt under platelet-derived growth factor-BB stimulation. Stellate cells were found to express methionine adenosyltransferase 2A. CONCLUSIONS: l-Cysteine and l-methionine regulate the activation of stellate cells. Their oral supply aids the suppression of the progression of liver fibrosis.     

Splenectomy ameliorates hepatic ischemia and reperfusion injury mediated by heme oxygenase-1 induction in the rat

BACKGROUND/AIMS: Ischemia/reperfusion (I/R) induces severe organic injury. I/R injury seems to be mainly caused by oxidative stress. The aim of this study was to determine the role of the spleen in experimental hepatic I/R injury in the rat. Stress protein heme oxygenase (HO)-1 plays a protective role against the oxidative injury. In normal state, HO-1 is highly expressed in the spleen. METHODS: Liver HO-1 expression was assessed by Western blot before and after splenects. Liver injury was assessed by measurement of ALT and AST and by histopathology. RESULTS: Although HO-1 was not detected in normal liver, levels of HO-1 protein gradually increased and peaked on 3 days after splenectomy. When splenectomy was performed 3 days prior to the hepatic (45-min) ischemia followed by (2-h) reperfusion, the levels of serum aspartate transaminase (AST) and alanine transaminase (ALT), as markers for hepatic injury, were improved compared to the rats with I/R alone. In addition, prior administration of zinc-protoporphyrin IX, a specific inhibitor of HO, suppressed the protective effect of the splenectomy on the subsequent hepatic I/R injury. Histopathological examination also confirmed these results. CONCLUSIONS: Our findings suggest that the elevated HO-1 levels by splenectomy play a protective role against hepatic I/R injury.     

Diffuse alveolar damage after inhalation of zinc oxide fumes]

A 57-year-old man with a 37-year occupational history of welding was admitted for high fever and dyspnea after inhalation of zinc oxide fumes during a period of welding without a protective mask. Chest radiography and CT showed bilateral diffuse ground-glass opacities, and blood gas analysis revealed that PaO2 was 48.1 torr in room air. A transbronchial lung biopsy was done, and revealed diffuse alveolar damage. We diagnosed the case as chemical pneumonia due to the inhalation of zinc oxide, and prescribed prednisolone 40 mg per day. As a result, his symptoms improved within several days. The inhalation of zinc oxide fume usually causes metal fume fever, but chemical pneumonia is also reported on rare occasions. As far as our examination of the literature has disclosed, this is the first report of diffuse alveolar damage after inhalation of zinc oxide fume.