The identification and characterization of a new GTP-binding protein (Gbp45) involved in cell proliferation and death related to mitochondrial function.

We describe the identification and characterization of a GTP-binding protein with a molecular weight of 45 kD (Gbp45). Gbp45 cDNA was found to overlap with a hypothetical human protein, PTD004, the sequence of which was previously deposited in the databases. The gene for PTD004 was recently found to be one of the ATPases, hOLA1 (human Obg-like ATPase 1).

Differences in molecular alterations of hepatocellular carcinoma between patients with a sustained virological response and those with hepatitis C virus infection.

Background/aims: The mechanism of hepatocarcinogenesis remains unclear in patients in whom hepatitis C virus (HCV) disappears after interferon (IFN) therapy. We compared molecular alterations in hepatocellular carcinoma (HCC) between patients with a sustained virological response (SVR) to IFN and patients with HCV.

Methods: The study group comprised 44 patients with HCV and 13 patients with SVR.

Mechanism of Liver Injury during Obstructive Jaundice: Role of Nitric Oxide, Splenic Cytokines, and Intestinal Flora.

To elucidate the roles of enteric bacteria and immunological interactions among liver, spleen and intestine in the pathogenesis of liver injury during obstructive jaundice, we studied the effects of antibiotics and splenectomy on bile-duct-ligated C57BL mice. When animals were subjected to bile-duct-ligation (BDL), plasma levels of bilirubin, alanine aminotransferase and aspartate aminotransferase increased markedly. However, the increases in plasma transaminases were significantly lower in splenectomized or antibiotics-treated groups than in the control BDL group.

[Oxidative stress and tissue injury].

Production of reactive oxygen species (ROS) and defense systems against them are balanced well in the living body. This balance is very important for the maintenance of physiological condition. The collapse of the balance such as in inflammation causes ROS toxicity and induces oxidative injury in various tissues.

Identification and characterization of endoplasmic reticulum-associated protein, ERp43.

Disposal of misfolded proteins from the lumen of the endoplasmic reticulum (ER) is one of the quality control mechanisms present in the protein secretory pathway. Through ER-associated degradation, misfolded substrates are targeted to the cytosol where they are degraded by proteasomes. Here we describe the identification of a human ER-associated 43-kD protein (ERp43) by sequencing of the subtraction suppression hybridization cDNA library from ER stress-treated cells.

Lack of mitochondrial DNA enhances growth of hepatocellular carcinoma in vitro and in vivo.

To elucidate the role of mitochondrial DNA (mtDNA) in determination of growth of hepatocellular carcinoma, we examined wild-type Hepa1-6 cells and their rho(0) cells with depleted mtDNA in vitro and in vivo. Cultured rho(0) cells grew more rapidly than did wild-type cells. Production of reactive oxygen species (ROS) was higher in wild-type cells than in rho(0) cells.

Production of reactive oxygen species in peripheral blood is increased in individuals with Helicobacter pylori infection and decreased after its eradication.

Background: Helicobacter pylori infection has been reported to cause gastroduodenal ulcer, mucosa-associated lymphoid tissue lymphoma, and gastric cancer. Recent studies have suggested that H. pylori infection may also associate with other diseases, including hematologic and dermatologic disorders, and cardiovascular injury, by unknown mechanisms.

Mannose-conjugated alendronate selectively depletes Kupffer cells and inhibits endotoxemic shock in the mice.

To elucidate the roles of Kupffer cells in the host-defense mechanisms and liver injury, we synthesized a mannose-conjugated alendronate (MANA) and examined its effects on Kupffer cells and lipopolysaccharide (LPS)-induced liver injury in the mice. Intravenous administration of a small amount of MANA (50mumol/kg) rapidly and selectively depleted Kupffer cells in the mice. The depletion of Kupffer cells by MANA resulted in a marked decrease of the production of both TNF-alpha and IL-1beta in the plasma during the liver injury induced by low (1mg/kg) and lethal (75mg/kg) doses of LPS.

L-carnitine suppresses the onset of neuromuscular degeneration and increases the life span of mice with familial amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal disease caused by progressive degeneration of motor neurons in the spinal cord and motor cortex. Although the etiology of ALS remains unknown, a mutation of the gene encoding Cu,Zn-superoxide dismutase (SOD1) has been reported in 20% of familial cases of ALS (FALS). Transgenic mice that overexpress a mutated human SOD1 exhibit a phenotype and pathology similar to those observed in patients with FALS.

A possible cooperation of SOD1 and cytochrome c in mitochondria-dependent apoptosis.

A small amount of reactive oxygen species (ROS) is generated through aerobic respiration even under physiological conditions. Because ROS are known to have various deteriorating actions, the way cells could evade the effects of ROS in and around mitochondria would determine the fate of cells. We previously reported that Cu,Zn-superoxide dismutase (SOD1), a cytosolic enzyme, is also localized in mitochondria in various types of cells.

Mitochondrial density determines the cellular sensitivity to cisplatin-induced cell death.

We studied the relationship between the mitochondrial density in the cells and the cellular sensitivity to the toxicity of cis-diaminedichloroplatinum II (cisplatin), a potent anticancer agent. Biochemical analyses revealed that the density of mitochondria in the intestinal epithelium changed markedly along its entire length. The density was the highest at the duodenum, medium at the jejunum, and the lowest at the ileum.

L-carnitine inhibits hypoglycemia-induced brain damage in the rat.

Hypoglycemia sometimes occurs in patients with diabetes mellitus who receive excessive doses of insulin. Severe hypoglycemia has been known to induce mitochondrial swelling followed by neuronal death in the brain. Since L-carnitine effectively preserves mitochondrial function in various cells both in vitro and in vivo, we investigated its effects on the neuronal damage induced by hypoglycemic insult in male Wistar rats.

Free radical theory of apoptosis and metamorphosis.

Reactive oxygen species (ROS) are the major factors that induce oxidative modification of DNA and gene mutation. ROS can elicit oxidative stress and affect a wide variety of physiological and pathological processes including embryonal development, maturation and aging.

Correlation between clinical characteristics and mitochondrial D-loop DNA mutations in hepatocellular carcinoma.

Background: Mitochondrial DNA (mtDNA) mutations are found in many kinds of human cancer. The aim of this study was to evaluate the relationship between mtDNA mutations in the liver and human hepatocarcinogenesis.

Methods: Direct sequencing of mtDNA was done in 54 hepatocellular carcinomas (HCCs) and 47 surrounding liver tissue samples, obtained from 54 patients with HCC, and in 5 liver samples without inflammation, obtained from 5 patients with metastatic liver tumors.

L-carnitine inhibits hepatocarcinogenesis via protection of mitochondria.

Hepatocellular carcinoma is usually preceded by chronic inflammation. However, the molecular mechanism in hepatocarcinogenesis is not well known. Recently, we reported that mitochondrial dysfunction plays an important role in hepatocarcinogenesis via the production of free radicals.

Inducible NO synthase inhibits the growth of free tumor cells, but enhances the growth of solid tumors.

To elucidate the role of nitric oxide (NO) in tumor cell growth in vivo, dynamic aspects of the growth of Ehrlich ascites tumor cells (EATCs) were studied in wild-type (WT) mice and in an inducible strain of NO synthase (iNOS)-deficient (iNOS(-/-)) mice. Kinetic analysis showed that the rate of free tumor cell growth in the peritoneal cavity was significantly higher in the iNOS(-/-) mice than in the WT mice. In contrast, EATCs inoculated subcutaneously rapidly grew and formed a solid tumor in WT mice, but failed to grow in iNOS(-/-) mice.

Edaravone inhibits acute renal injury and cyst formation in cisplatin-treated rat kidney.

Background: Although cis-diamminedichloroplatinum (II) (cisplatin) is an effective anticancer agent, its clinical use is highly limited predominantly due to its adverse effects on renal functions. The present work examined the therapeutic potential of edaravone, a free radical scavenger, for inhibiting cisplatin-induced renal injury.

Methods: Edaravone, 3-methyl-1-phenyl-pyrazolin-5-one, was administrated intravenously at a dose of 30 mg/kg of body weight to male Wistar rats (200-220 g).

Effect of endogenously generated nitric oxide on the energy metabolism of peritoneal macrophages.

To understand the role of nitric oxide (NO) in the regulation of cellular metabolism in peritoneal macrophages under physiological low oxygen tension, its effect on the respiration and energy metabolism was examined with casein-induced peritoneal macrophages from the rat. Intraperitoneal injection of casein transiently induced peritoneal infiltration of neutrophils (peaked on day 1) followed by the migration of macrophages that peaked on day 2. Western blotting analysis using antibodies against inducible type of NO synthase (iNOS) revealed that macrophages appeared in the peritoneal cavity during an early stage (approximately day 2) but not during the late stage (day 3 approximately) of inflammation expressed iNOS and generated substantial amounts of NO by a mechanism that was inhibited by N-iminoethyl-L-ornithine (NIO), a specific inhibitor of iNOS.

Mitochondrial generation of reactive oxygen species and its role in aerobic life.

Mitochondria are the major site for the generation of ATP at the expense of molecular oxygen. Significant fractions (approximately 2%) of oxygen are converted to the superoxide radical and its reactive metabolites (ROS) in and around mitochondria. Although ROS have been known to impair a wide variety of biological molecules including lipids, proteins and DNA, thereby causing various diseases, they also play critical roles in the maintenance of aerobic life.

Cross talk of nitric oxide, oxygen radicals, and superoxide dismutase regulates the energy metabolism and cell death and determines the fates of aerobic life.

Although oxygen is required for the energy metabolism in aerobic organisms, it generates reactive oxygen and nitrogen species that impair a wide variety of biological molecules, including lipids, proteins, and DNA, thereby causing various diseases. Because mitochondria are the major site of free radical generation, they are highly enriched with enzymes, such as Mn-type superoxide dismutase in matrix, and antioxidants including GSH on both sides of inner membranes, thus minimizing oxidative stress in and around this organelle. We recently showed that a cross talk of nitric oxide and oxygen radicals regulates the circulation, energy metabolism, reproduction, and remodeling of cells during embryonic development, and functions as a major defense system against pathogens.

Multicentric occurrence of hepatocellular carcinoma in patients with a somatic mutation of mitochondorial DNA and hepatitis C virus.

The relationship between the multicentric occurrence of hepatocellular carcinoma (HCC) and the frequency of mutation of mitochondrial DNA (mtDNA) in the noncancerous hepatic tissue in patients infected with hepatitis C virus was investigated. Of the 48 patients, multicentric occurrence of HCC was found in ten of 33 patients with three or more mutations in the mtDNA, whereas no patients had multicentric HCCs in 15 patients with two or fewer mutations in the mtDNA (P=0.0201).

Mice lacking inducible nitric oxide synthase show strong resistance to anti-Fas antibody-induced fulminant hepatitis.

Although nitric oxide (NO) plays important roles in pathogenesis of various liver diseases, the role of NO in the in vivo mechanism of Fas-mediated fulminant hepatitis is not known well. The effect of anti-Fas antibody (Jo2) on the survival, liver function, and histology was analyzed in wild-type (WT) and inducible NO synthase (iNOS)-deficient (iNOS(-/-)) mice. Upon intravenous injection of a lethal dose of Jo2, WT mice died on fulminant hepatitis within 12h.

L-Carnitine inhibits cisplatin-induced injury of the kidney and small intestine.

Although cis-diamminedichloroplatinum (II) (cisplatin) is a potent anticancer drug, clinical use of this agent is highly limited predominantly because of its strong side effects on the kidney and gastrointestinal tracts. We found that cisplatin impaired respiratory function and DNA of mitochondria in renal proximal tubules and small intestinal mucosal cells, thereby inducing apoptosis of epithelial cells. Cisplatin-induced mitochondrial dysfunction and DNA (mtDNA) injury, lipid peroxidation, and apoptosis of epithelial cells in the kidney and small intestine were strongly inhibited by L-carnitine.