The present study was designed to investigate whether acute ethanol intoxication increases the production of active oxidants, and subsequently promotes apoptosis of hepatocytes. Hepatocytes were ...isolated from male Wistar rats, and cultured in the presence or absence of ethanol. The fluorescence in situ nick end labeling method and an enzyme‐linked immunosorbent assay (ELISA) system to quantify fragmented DNA were used to estimate apoptotic change in hepatocytes. Nuclear morphological alterations and membrane barrier dysfunction of hepatocytes were assessed by staining with Hoechst 33342 and propidium iodide (PI). Intracellular glutathione level was determined as the fluorescence of monochlorobimane (MCLB), which forms conjugate with glutathione to become fluorescent. Ethanol (100 mmol/L) increased the amount of fragmented DNA and the number of apoptotic hepatocytes in vivo as well as in vitro. These ethanol‐induced alterations in hepatocytes were attenuated by simultaneous incubation with either 4‐methylpyrazole, an inhibitor of alcohol dehydrogenase, or dimethylthiourea, an intracellular oxidant scavenger. Diethyl maleic acid (DMA), a glutathione depletor, enhanced the induction of apoptotic change, and decreased membrane barrier function in ethanol‐treated hepatocytes, whereas ethanol per se did not increase the number of PI‐positive hepatocytes. Furthermore, combination of ethanol and DMA but not ethanol alone decreased the hepatocyte MCLB fluorescence. Taken together, the present study suggests that active oxidants produced during ethanol metabolism mediate fragmentation of DNA in hepatocytes, and that intracellular antioxidants such as glutathione play a critical role in the cytoprotective mechanisms of hepatocyte against lethal cell death, ie, apoptosis, induced by ethanol.
ABSTRACT
Satellite cells, resident myogenic stem cells found in postnatal skeletal muscle, are most abundant during early postnatal development and sharply decline in frequency thereafter to adult ...levels in mice and rats. Therefore, postnatal changes in satellite cell mitotic activities are important aspects for further understanding a muscle growth strategy. In large meat‐production animals, however, the traditional in vivo proliferation assay may be less realistic because it requires intra‐peritoneal (ip) injection of huge dosage of mutagenic nucleosides, 3H‐labeled thymidine or bromodeoxyuridine (BrdU), at each age‐time of sacrifice. We report in the present pilot study using rats that in vivo proliferation activity of satellite cells can be evaluated by an in vitro BrdU‐incorporation assay in early cultures. Briefly, satellite cells were prepared from upper hind‐limb and back muscles and maintained for 24 h with imposing by BrdU addition for the last 2 h, followed by the regular immunocytochemistry for determining BrdU‐incorporated cell percentage. This in vitro assay demonstrated a rapid decrease in proliferating satellite cell frequency to the adult level during about 3‐month period after birth, and yielded a high correlation to the measurements by the in vivo BrdU ip‐injection method during the postnatal period examined from day‐2 to month‐11. The in vitro proliferation assay may be further adaptable for large domestic animals by the combination with a muscle biopsy technique that enables age‐interval sampling from the same growing animals.
Purpose: Childhood epilepsy with occipital paroxysms (CEOP) is characterised by ictal visual hallucinations and occipital epileptiform activity on interictal EEG. A variant has been described with ...nonvisual symptoms including tonic head and eye deviation, vomiting, and episodes of partial status epilepticus. We fully documented the electroclinical features of such patients to determine whether classification separate from CEOP is justified.
Methods: This was a multicentre study with participating investigators submitting details of patients with idiopathic occipital seizures characterised by ictal head or eye deviation and vomiting.
Results: One hundred thirteen patients were recruited. Seizures began in early childhood (mean, 4.6 years) and occurred infrequently (mean total seizures, 3); 30% of patients had only a single seizure. Two thirds of seizures were nocturnal. Ictal eye deviation occurred in 79%, vomiting in 70%, and head deviation in 35%. Seizures were predominantly complex partial in type. Partial status epilepticus occurred in 44% of patients. Seventy‐four percent of patients had occipital interictal EEG epileptiform activity, predominantly right sided, with fixation‐off sensitivity. Extraoccipital EEG abnormalities occurred in 35% of patients. Prognosis was excellent: the mean duration of active seizures was 1 year.
Conclusions: Although the two groups shared identical EEG features, the distinct clinical symptoms probably justify separate classification. Early‐onset benign occipital seizure syndrome (EBOSS) is suggested as an appropriate name for the variant group.
The charge-transfer compound {Ru2(O2CPh-o-Cl)4}2TCNQ(MeO)2·CH2Cl2 (1; o-ClPhCO2 − = o-chlorobenzoate; TCNQ(MeO)2 = 2,5-dimethoxy-7,7,8,8-tetracyanoquinodimethane) was synthesized from the reaction of ...the neutral precursors Ru2 II,II(O2CPh-o-Cl)4 (abbreviated as Ru2 II,II or Ru2 4+) and TCNQ(MeO)2 in a CH2Cl2/nitrobenzene solution. The structure consists of two-dimensional layers consisting of an infinite array in which Ru2 II,II units are involved in charge transfer to TCNQ(MeO)2 to give a formal charge of {Ru2 4.5+}-TCNQ(MeO)2 • −-{Ru2 4.5+}. Interstitial CH2Cl2 molecules are located in the void spaces between the layers. Strong intralayer magnetic coupling between the units Ru2 II,II with S = 1 or Ru2 II,III with S = 3/2 and TCNQ(MeO)2 • − with S = 1/2, as well as long-range ordering due to antiferromagnetic interlayer interactions, was observed. An antiferromagnetic ground state exists below T N = 75 K, which undergoes a metamagnetic transition under applied fields less than 2 T to a field-induced canted antiferromagnetic state with large coercivities up to H c = 1.6 T at 1.8 K. Compound 1 gradually loses the interstitial CH2Cl2 molecule at room temperature to form a dried sample (1-dry) without loss of crystallinity and converts nearly reversibly back to 1 after being exposed to CH2Cl2 vapor for 72 h (distinguished as 1′). Interestingly, during this process there is no significant change in lattice dimensions and bond distances or angles with a volume change of only 1.2 vol %. The only discernible difference is ordering/disordering of a pendant ligand orientation, but the magnetism is dramatically altered to a ferromagnetic state with T c ≈ 56 K for 1-dry. The magnetic property changes are gradual and depend on the degree of interstitial CH2Cl2 molecule loss with reversibility in the process of going between 1 and 1-dry. In addition, in the case of partially desolvated crystals that have mixed domains of ferromagnetically and antiferromagnetically ordered domains for desolvated and solvated segments, respectively, the complete change to ferromagnet can also be triggered by magnetic fields even if the desolvated segments are comparatively minor compared to the solvated segments in a crystal. Surprisingly, the information of the existence of ferromagnetically ordered domains is dynamically recorded in the entire crystal after applying significant magnetic fields as if the majority of the antiferromagnetically ordered domains for solvated segments were never present.
Six terpenoids have been isolated from the root bark of
Tripterygium hypoglaucum, along with 14 known compounds. The structures of the terpenoids were elucidated as 3,11,14-oxo-abieta-8,12-diene, ...3β-hydroxy-12,14-dimethoxyabieta-8,11,13-triene, 3β-hydroxy-11α-ethoxyurs-12-ene, 3β-hydroxy-11α-methoxyurs-12-ene, 3β-hydroxy-11α-methoxyolean-12-ene-28-oic acid, and 1β-benzoyl-8α-cinnamoyl-4α,5α-dihydroxydihydroagarofuran.
Three C-17 diacetylenic compounds (
1–
3), one monoterpenoid (
4), seven ceramides (
5a–
g), seven cerebrosides (
6a–
f) and nine known compounds were isolated from the methanolic extract of
...Hydrocotyle leucocephala. The isolated compounds were active in the LPS induced cytokine production assay for IL-10, IL-12, and TNF-α.
Three C-17 diacetylenic compounds (
1–
3), one monoterpenoid (
4), seven ceramides (leucoceramides A–G,
5a–
g), six cerebrosides (leucocerebrosides A–F,
6a–
f) and nine known compounds were isolated from the methanolic extract of
Hydrocotyle leucocephala. Their structures were established by spectroscopic methods. The isolated compounds
1–
3,
5a–
g,
6a–
f and
7 were shown to be active in the lipopolysaccharide (LPS) induced cytokine production assay for IL-10, IL-12, and TNF-α.
Two new diterpenes, named sibiriquinone A (1) and B (2), along with four known diterpenes have been isolated from the aerial part of Veronicastrum sibiricum. Their structures were elucidated by ...spectroscopy. The isolated compounds showed significant immunosuppressive activities.
Helicobacter pylori infects over half the population worldwide and is a leading cause of chronic gastritis and gastric cancer. However, the mechanism by which this organism induces inflammation and ...carcinogenesis is not fully understood. In the present study we used insulin-gastrin (INS-GAS) transgenic mice that fully develop gastric adenocarcinoma after infection of H. pylori-related Helicobacter felis. Histological examination revealed that more than half of those mice developed invasive adenocarcinoma after 8 months of infection. These carcinomas were stained by NCC-ST-439 and HECA-452 that recognize 6-sulfated and non-sulfated sialyl Lewis X. Lymphocytic infiltration predominantly to submucosa was observed in most H. felis-infected mice, and this was associated with the formation of peripheral lymph node addressin (PNAd) on high endothelial venule (HEV)-like vessels detected by MECA-79. Time-course analysis of gene expression by using gene microarray revealed upregulation of several inflammation-associated genes including chemokines, adhesion molecules, surfactant protein D (SP-D), and CD74 in the infected stomach. Immunohistochemical analysis demonstrated that SP-D is expressed in hyperplasia and adenocarcinoma whereas CD74 is expressed in adenocarcinoma in situ and invasive carcinoma. These results as a whole indicate that H. felis induces HEV-like vessels and inflammation-associated chemokines and chemokine receptors, followed by adenocarcinoma formation.
Cholesterol efflux (CE) is the initial and important step of reverse cholesterol transport (RCT), a major protective system against atherosclerosis. However, most of the molecular mechanism for CE ...still remains to be clarified. In the present study, cDNA subtraction revealed that the expression of a member of the Rho GTPase family, Cdc42Hs, was markedly decreased in both passaged fibroblasts and macrophages (Mφ) from patients with Tangier disease (TD), a rare lipoprotein disorder with reduced CE. This small G protein is known to have many cell biological activities such as rearrangement of actin cytoskeleton and vesicular transport, however the association between this molecule and lipid transport has never been reported. We demonstrate that MDCK cells expressing the dominant negative form of Cdc42Hs had reduced CE, inversely ones expressing the dominant active form had increased CE. From these observations, we would like to raise a novel hypothesis that this type of small G protein may play a role in some steps of CE. To our knowledge, the present study is the first demonstration that the expression of this molecule is altered in cells from human disease.
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