Relation of Poor Nutritional Status to Mild Cognitive Impairment in Patients with Coronary Artery Disease.

OBJECTIVES: Nutritional status affects cerebral circulation and cognitive function. More attention needs to be paid to nutritional status in coronary artery disease (CAD) patients, yet the relation between nutritional status or dietary intake (DI) and cognitive function or mild cognitive impairment (MCI) in CAD patients remain unclear. Thus, we examined the following relations: 1) that between nutritional status and cognitive function, and MCI and 2) that between DI and cognitive function, and MCI. DESIGN, SETTING, AND PARTICIPANTS: We conducted a cross-sectional study of 208 patients with CAD but without dementia. MEASUREMENTS: MCI was estimated with the Japanese version of the Montreal Cognitive Assessment (MoCA-J). Nutritional status was assessed by the Geriatric Nutritional Risk Index (GNRI), and DI was assessed by total energy intake per day. We investigated the relation between nutritional status or DI and cognitive function by Pearson correlation analysis, and that between nutritional status or DI and MCI by multivariable logistic regression analysis. RESULTS: The GNRI and DI were positively associated with the MoCA-J score (r = 0.23, p < 0.001, and r = 0.24, p < 0.001, respectively), and both were independently associated with MCI in the multivariable logistic regression analysis (odds ratio, 0.96; p = 0.045, and odds ratio, 0.998; p = 0.020, respectively). CONCLUSIONS: Poor nutritional status and low DI were found to be significantly associated with cognitive function and MCI in CAD patients. Our findings regarding nutritional status and DI might be useful for clinicians to prevent or intervene in the early cognitive decline of inpatients with CAD.

Metal-ion-induced expression of gene fragments from subseafloor micro-organisms in the Kumano forearc basin, Nankai Trough.

AIMS: Using substrate-induced gene-expression (SIGEX) screening on subseafloor sediment samples from the Nankai Trough, Japan, we identified gene fragments showing an induction response to metal ions. METHODS AND RESULTS: Environmental DNA libraries in Escherichia coli host cells were tested by the addition of metal ions (Ni2+ , Co2+ , Ga3+ or Mo6+ ), followed by cell sorting of clones exhibiting green fluorescence upon co-expression of green fluorescence protein downstream of the inserted gene fragments. One clone displayed Ni2+ -specific induction, three clones displayed Ga3+ -specific induction and three clones displayed an induction response to multiple metal ions. DNA sequence analysis showed that a variety of genes showed induction responses in the screened clones. CONCLUSIONS: Using the SIGEX approach, we retrieved gene fragments with no previously identified response to metal ions that exhibited metal-ion-induced expression. This method has the potential to promote exploration of gene function through gene-induction response. SIGNIFICANCE AND IMPACT OF THE STUDY: We successfully linked gene-induction response with sequence information for gene fragments of previously unknown function. The SIGEX-based approach exhibited the potential to identify genetic function in unknown gene pools from the deep subseafloor biosphere, as well as novel genetic components for future biotechnological applications.

Expression of activation-induced cytidine deaminase in human hepatocytes via NF-kappaB signaling.

Activation-induced cytidine deaminase (AID) is involved in somatic DNA alterations of the immunoglobulin gene for amplification of immune diversity. The fact that constitutive expression of AID in mice causes tumors in various organs, including lymphoid tissues and lungs, suggests the important role of the aberrant editing activity of AID on various tumor-related genes for carcinogenesis. AID expression, however, is restricted to activated B cells under physiological conditions. We demonstrate here that ectopic AID expression is induced in response to tumor necrosis factor-alpha stimulation in cultured human hepatocytes. The proinflammatory cytokine-mediated expression of AID is achieved by IkappaB kinase-dependent nuclear factor (NF)-kappaB signaling pathways. Hepatitis C virus, one of the leading causes of hepatocellular carcinoma (HCC), enhanced AID expression via NF-kappaB activation through expression of viral core protein. The aberrant expression of AID in hepatoma-derived cells resulted in accumulation of genetic alterations in the c-myc and pim1 genes, suggesting that inappropriate expression of AID acts as a DNA mutator that enhances the genetic susceptibility to mutagenesis in human hepatocytes. Our current findings indicate that the inappropriate expression of AID is induced by proinflammatory cytokine stimulation and may provide the link between hepatic inflammation and the development of HCC.

Effects of long-term administration of sulindac on APC mRNA and apoptosis in colons of rats treated with azoxymethane.

PURPOSE: Non-steroidal anti-inflammatory drugs, including sulindac, have been shown to exhibit anti-colon cancer activity; however, the detailed mechanisms concerning continuous long-term administration are still unclear. Therefore, we examined the anti-colon carcinogenesis effects of sulindac after prolonged administration. METHODS: Administration of AOM, a colon-specific carcinogen, induced colonic preneoplastic lesions, which can progress to carcinomas about 40-50 weeks after AOM administration. We studied the effects of sulindac on the incidence of preneoplastic lesions, proliferative activity of colonic cells (AgNORs), tumor suppressor adenomatous polyposis coli (APC) gene expression, and apoptosis using AOM-treated rat colon mucosa at 4 weeks and 40 weeks (early and late stage of colon carcinogenesis, respectively). RESULTS: Sulindac suppressed the development of preneoplastic lesions induced by AOM at 4 weeks and 40 weeks by about 50% ( P<0.01); the proliferative activity of colonic cells increased by AOM was suppressed almost completely. Furthermore, APC expression was significantly increased by sulindac at both the early and late stages ( P<0.01). However, apoptosis was clearly increased at the early stage ( P<0.01), but not at the late stage. CONCLUSIONS: APC overexpression induced by sulindac can suppress colon carcinogenesis at both the early and late stages, but apoptosis might work as one of anti-cancer mechanisms at the early stage of colon carcinogenesis.

Molecular changes in the early stage of colon carcinogenesis in rats treated with azoxymethane.

To elucidate early molecular events related to colon carcinogenesis, we examined alterations in the expression of colon cancer-related genes such as cyclooxygenase (COX)-2, APC and c-Myc, cell proliferation and apoptosis in the background colon mucosa, and K-ras mutation at aberrant crypt foci (ACF) in the colons of azoxymethane (AOM)-treated rats 4 weeks after the first exposure to AOM. About 40 ACF/colon were induced in the colons of rats treated with AOM (Group 1); however, rats not treated with AOM (Group 2) showed no ACF formation in the colon. The level of AgNORs in the colonic mucosa was significantly higher in Group 1 than in Group 2 (P<0.01). The colonic mucosa in Group 1 looked macroscopically and histologically normal, but the proliferative activity of the mucosa of rats treated with AOM was clearly elevated. COX-2 mRNA expression was not detected in normal colonic mucosa in Group 2, but 3 out of 10 rats in Group 1 showed COX-2 mRNA expression in their colons by reverse transcription (RT)-polymerase chain reaction (PCR). There was a tendency toward an increased expression level of COX-2 in the AOM-treated group. The level of APC mRNA expression in Group 1 was significantly lower than that in Group 2 (P<0.01). Moreover, the level of c-Myc mRNA expression in Group 1 was significantly higher than that in Group 2 (P<0.01). An average of 0.034+/-0.006% apoptosis in colonic mucosa was detected in Group 1; the incidence of apoptosis in Group 2 was 0.021+/-0.005%. The difference between Groups 1 and 2 was significant (P<0.01). These results indicate that apoptosis was possibly induced to eliminate cells damaged by AOM administration. Six out of 22 (27%) ACF with 4 or more crypts showed K-ras mutations at codon 12; all mutations were G to A transitions (GGT to GAT). ACF with 1-3 crypts showed no mutations in the K-ras gene. In conclusion, AOM caused an increase in COX-2 and c-Myc mRNA expression, a decrease in APC mRNA expression, induction of apoptosis in normal-appearing colonic mucosa, and a K-ras mutation in ACF with 4 or more crypts. These findings may help to identify key targets in the early steps of colon carcinogenesis, against which drugs that would be broadly effective for chemoprevention of colon cancer could be developed.

Comparison of the kinetic disposition of and serum gastrin change by lansoprazole versus rabeprazole during an 8-day dosing scheme in relation to CYP2C19 polymorphism.

BACKGROUND: Little is known about differences in the disposition kinetics and pharmacological effects on gastrin levels between lansoprazole and rabeprazole given in a repeated dosing scheme with respect to the polymorphic CYP2C19. AIM: To provide preliminary information that should be considered when prescribing proton-pump inhibitors (PPIs) for the treatment of acid-related diseases with reference to the CYP2C/9 genotypic status. METHODS: Helicobacter pylori-negative healthy volunteers were divided into the following three groups (n = 5 each) on the basis of genotyping for CYP2C19: homozygous (hmEMs) and heterozygous extensive metabolizers (htEMs), and poor metabolizers (PMs). All received once-daily 30-mg doses of lansoprazole or 10-mg doses of rabeprazole during an 8-day course in a crossover manner. RESULTS: The relative values for the area under the serum concentration-time curve (AUC) of lansoprazole and rabeprazole in the hmEMs, htEMs, and PMs after the final doses were 1:1.7:3.9 and 1:1.7:3.8, respectively. The relative AUCs of gastrin in the hmEMs, htEMs, and PMs were 1.6:2.6:3.1 for lansoprazole and 1.6:2.6:2.9 for rabeprazole, respectively. CONCLUSION: The disposition kinetic behavior of the two PPIs is co-segregated with CYP2C19. The magnitude of CYP2C19-dependent drug availability in the systemic circulation and resulting gastrin response appears to be fairly similar between the two drugs within the same CYP2C19 genotypic groups after a multiple-dosing regimen.

Loss of heterozygosity of the mannose 6-phosphate/insulin-like growth factor II receptor and p53 genes in human hepatocellular carcinoma.

Information about M6P/IGF2R and p53 genes in hepatocarcinogenesis is limited and controversial. We tested the loss of heterozygosity (LOH) of M6P/IGF2R and p53 genes in cirrhotic and neoplastic foci in surgically resected livers of 30 patients with hepatocellular carcinoma (HCC). The DNAs extracted from microdissected specimens were used for polymerase-chain-reaction (PCR)-based assay. LOH of the M6P/IGF2R gene in the primary HCCs was detected in 10 of 22 informative cases (45%). In five of these 10 cases (50%), LOH was detected in cirrhotic lesions adjacent to the HCCs. The allelic loss patterns of M6P/IGF2R in liver cirrhosis (LC) were identical to those in the corresponding HCC, suggesting that HCC could develop from one of the cells in which M6P/IGF2R had been lost. Furthermore, LOH of the p53 gene in HCC was detected in 10 (43%) of 23 informative cases, and p53 loss in cirrhotic foci adjacent to HCC was shown in one of the 10 cases (10%). The pattern of allelic loss of the p53 gene in the cirrhotic foci was identical with that in the corresponding tumor. The LOH of the M6P/IGF2R and p53 genes occurred independently in HCCs. LOH of the M6P/IGF2R locus was a relatively frequent and possibly early event in hepatocarcinogenesis, and LOH of the M6P/IGF2R gene and LOH of the p53 gene occurred independently.

Influence of angiotensin II type 1 receptor polymorphism on hypertension in patients with hypercholesterolemia.

Essential hypertension results from the combined influence of environmental and genetic factors. The relationship between angiotensin II type 1 receptor (AT(1)) A-C(1166) polymorphism and essential hypertension is controversial. Because it is accepted that high concentration of serum cholesterol is one of risk factors of atherosclerosis, we investigated the influence of the AT(1) A-C(1166) polymorphism on hypertension in patients with hypercholesterolemia. A total of 131 hypertensive, 97 borderline, and 175 normotensive subjects were enrolled in this study. We selected hypercholesterolemic subjects on the condition that their serum concentration of total cholesterol was >220 mg/dl, and obtained 55 hypertensive, 24 borderline, and 52 normotensive subjects with hypercholesterolemia. There were no significant differences in the genotype nor allele frequency between hypertensive and normotensive subjects in the overall population. However, the presence of the C allele of the AT(1) gene has a tendency to increase the value of systolic blood pressure not only in subjects with hypercholesterolemia but also in the overall population. Furthermore, we found a significant relationship between the AT(1) polymorphism and hypertension in subjects with hypercholesterolemia; i.e., the frequency of the C allele of the AT(1) gene was significantly higher in hypertensives than in normotensives (P<0.005). These results suggested that high concentration of total cholesterol was an important risk factor to the occurrence of essential hypertension for patients who carried the C allele of the AT(1) gene.

Sulindac and a cyclooxygenase-2 inhibitor, etodolac, increase APC mRNA in the colon of rats treated with azoxymethane.

BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) have been reported to protect against the development of colon cancer. However, the mechanism(s) by which NSAIDs exert their effects is not clear. AIMS: The aim of this study was to examine the effects of NSAIDs on mRNA expression of tumour suppressor adenomatous polyposis coli (APC) gene in rat colon mucosa. METHODS: Starting at six weeks of age, three groups of rats (groups 1, 2, and 3) were treated with azoxymethane (AOM), a colon specific carcinogen, and another three groups (groups 4, 5, and 6) were not given AOM. Groups 2 and 3 were given 10 mg/kg of sulindac or etodolac, respectively, three times weekly during the experiment. Groups 4 and 5 were also given sulindac or etodolac, respectively, in the same manner as in groups 2 and 3. Group 6 (untreated control) was not given any agent (AOM or NSAIDs). At 10 weeks of age, preneoplastic lesions (aberrant crypt foci (ACF)) induced by AOM in the colon were counted, and the level of expression of APC mRNA in the colonic mucosa was estimated by the reverse transcription-competitive polymerase chain reaction method and northern blot analysis. RESULTS: Mean occurrence of ACF in rats in groups 2 and 3 was reduced to approximately 50% of that in group 1. The level of APC mRNA expression in group 1 (AOM alone) was lower than that in group 6 (untreated control) (p<0.05); however, levels of APC mRNA expression in groups 2, 3, 4, and 5, to which NSAIDs had been administered, were significantly increased compared with levels in groups 1 and 6 (p<0.01). CONCLUSIONS: Both sulindac and etodolac reduced the occurrence of ACF and induced an increase in APC mRNA in rat colon mucosa.

Spontaneous and cytokine regulated c-fos gene expression in rheumatoid synovial cells: resistance to cytokine stimulation when the c-fos gene is overexpressed.

OBJECTIVE: To study the effect of cytokines on the transactivation of the c-fos gene in relation to the contribution of overexpression of c-fos/AP-1 in rheumatoid joint destruction. METHODS: The promoter region (-447 to +109) of the human c-fos gene was integrated upstream of the chloramphenicol acetyltransferase (CAT) reporter gene, and the effect of cytokines on the expression of the c-fos gene was studied in the rheumatoid synovial cells of early (3-4) or late (14-18) passages, in the presence or absence of cytokines, by the transient transfection assay. RESULTS: Expression of c-fos gene was enhanced by tumour necrosis factor alpha (TNF alpha) and interleukin 6 (IL6) in the synovial cells of early passage, whereas it was not enhanced in the synovial cells of late passage. The c-fos gene expression was also enhanced by 13-O-tetradecanoyl phorbol-13-acetate (TPA) in early passage but was somewhat suppressed in the late passage. It was found that the c-fos gene and c-Fos protein were both increased in the synovial cells of late passage. Similarly, c-fos gene expression was also not increased by TPA or cytokine stimulation in the stable c-fos transformants (fos-pH8) or H-ras transformed NIH3T3 cells (NIH H-ras cells) that constitutively expressed c-fos genes. CONCLUSIONS: Although TNF alpha and IL6 augmented c-fos gene expression of rheumatoid synovial cells, transactivation of c-fos gene became resistant against cytokine stimulation under prolonged expression of c-fos gene, which may impart a tumour-like characteristic to rheumatoid synovial cells.

Effects of trimebutine maleate on delayed rectifier K+ currents in guinea-pig ventricular myocytes.

The effects of trimebutine maleate, a drug commonly used to regulate motility in the gastrointestinal tract, on the delayed rectifier K+ current (I(K)) were evaluated in guinea-pig ventricular myocytes to determine whether the drug has a proarrhythmic effect through blockade of I(K). Trimebutine decreased I(K) in a concentration-dependent manner. To investigate the effects of trimebutine on two components of I(K) (I(Kr) and I(Ks); rapidly activated and slowly activated components, respectively), we performed the envelope-of-tails test. Trimebutine-sensitive I(K) was determined by digital subtraction of I(K) during exposure to trimebutine from control I(K) for each duration of the test pulse over the range 50 ms-2 s. The ratio of deltaI(K,tail)/deltaI(K) plotted against pulse duration for trimebutine-sensitive I(K) gradually decreased to a steady-state value as the duration of the test pulse was lengthened. This finding suggested a weak inhibitory effect of trimebutine on both I(Kr) and I(Ks). The effects of trimebutine on the inward rectifier K+ current (I(K1)) responsible for the resting potential and final repolarization phase of the action potential were investigated by applying voltage clamp ramps over a broad range of potentials. No significant effects were observed at 10 or 100 microM. We next investigated the effects of the drug on the L-type Ca2+ current (I(Ca)). Significant inhibition of I(Ca) was observed at trimebutine concentrations greater than 10 microM. These results suggested that trimebutine maleate has weak inhibitory effects on I(Kr), I(Ks) and I(Ca) at concentrations much higher than those in clinical use.

Disopyramide decreases the fasting serum glucose level in man.

The effect of disopyramide, a class Ia antiarrhythmic drug, on the serum glucose level was evaluated in 6 consecutive in-patients. A 19-hour starvation test was repeated with oral administration of sustained-release disopyramide (150 mg) 0 and 12 hours after starting the test. Serum glucose levels during the starvation test decreased with disopyramide administration from a mean value of 96.5 +/- 1.8 to 85.9 +/- 1.4 mg/dl (24 samples, p < 0.05). The average reduction of the serum glucose level by disopyramide in each patient was 9.7 +/- 2.2 mg/dl. The decrease in the serum glucose level was not related to the serum concentration of disopyramide or serum creatinine levels. The decrease in the serum glucose level was larger in older patients (r = 0.75) and in light patients under 45 kg. These results suggested that disopyramide reduced the fasting serum glucose levels even in normal ranges as a common side effect of the drug, and that not only the occurrence of severe hypoglycemia but also the decrease in glucose levels were influenced by multiple factors including age and body weight.

Effects of itopride hydrochloride on the delayed rectifier K+ and L-type CA2+ currents in guinea-pig ventricular myocytes.

The effects of itopride hydrochloride, a new drug used to regulate motility in the gastrointestinal tract, on the delayed rectifier K+ current (I(K)) and the L-type Ca2+ current (I(Ca)) were evaluated in guinea-pig ventricular myocytes at concentrations of 1, 10 and 100 microM to determine whether the drug has a proarrhythmic effect through blockade of I(K). Itopride did not affect I(K) at concentrations of 100 microM or less, and no significant effects of 1, 10 or 100 microM itopride were observed on the inward rectifier K+ current (I(K1)) responsible for the resting potential and final repolarization phase of the action potential. We next investigated the effects of itopride on L-type Ca2+ current (I(Ca)). Significant inhibition of I(Ca) was observed at itopride concentrations greater than 10 microM. These results suggested that itopride hydrochloride has an inhibitory effect on I(Ca) at concentrations much higher than those in clinical use.

[Growth regulation of sex steroid hormone in endometrial carcinoma transplanted into nude mice].

By using ovariectomized nude mice, the hormone reactivity of endometrial carcinoma was evaluated. HEC-88nu cultured cells originated from human endometrial carcinoma were first transplanted to the animal in each experiment. Estrogen receptor (ER) of HEC-88nu reveals positive originally. Hormone pellets containing medroxyprogesterone acetate (MPA) and 17 beta-estradiol (E2) were used. The results were as follows: 1. The proliferation of this tumor was accelerated remarkably by administration of E2 pellet. 2. By administration of MPA pellet, the proliferation was inhibited from the beginning but progressed flatly afterwards maintaining 50% of the control. 3. When MPA was administered upon priming the tumor with E2, the proliferation began to be inhibited after 2 weeks developing 60% of suppression 5 weeks later. 4. Progesterone receptor (PR) of the tumor was induced starting at week 2 when E2 was given and revealed 189 fmol/mgP at week 5. 5. As the morphological changes due to hormone, light eosin-stainability, rarefaction and swelling of the cytoplasms were the common characteristics. 6. It was suggested that both hormonal and pharmacological actions take part in the mechanism of progestin to act on endometrial carcinoma.

[Diagnosis and treatment of dysplasia of the uterine cervix].

The accuracy of diagnosis and the results of treatment for dysplasia of the uterine cervix are analyzed. Three hundred and thirty-seven cases of mild, 231 of moderate and 200 of severe dysplasias have been treated at Kitasato University Hospital. By cytology 40.4% of mild, 74.5% of moderate and 89.0% of severe dysplasias were judged suspicious or positive, and the accurate cytopathological detection rate was 19.8%, 16.1% and 60.1%, respectively. Abnormal colposcopic findings such as M, P and W were observed in more than 95% of cases with dysplasia. The area of abnormal findings tends to be large as the lesion becomes more severe. Mild dysplasia progressed to cervical cancer in 1.5%, moderate dysplasia in 2.4% and severe dysplasia in 9.4%. Conservative therapy by laser vaporization or conization was used for 31 mild, 96 moderate and 114 severe dysplasias. The cure rate of each dysplasia was 100%, and pregnancy following laser therapy was encountered in 28 cases without abnormal course. It is concluded that colposcopy is more useful in diagnosis of mild and moderate dysplasia than cytology, and conservative laser treatment is an effective method for dysplasia.

[Establishment and characterization of a new SCC antigen producing cell line (HCS-2) from a carcinoma of the uterine cervix].

The cell line designated HCS-2 established from a squamous cell carcinoma of the uterine cervix has been subcultivated 77 times since Nov. 8, 1983. The cultured cells appear epithelial in shape, with a pavement-like arrangement and grow without contact inhibition. In electron microscopy, the cells are characterized by desmosomal cell contacts and a few tonofilaments. The cells are transplanted subcutaneously to nude mice and produce tumor which resembles the original tumor of large cell non-keratinizing squamous cell carcinoma. The growth rate of subculture has increased gradually, and population doubling time of cells at 17th passage was about 65 hours. The chromosome studies show aneuploidy and chromosomal number was mainly from hypertriploid to hypotetraploid range. The modal number of cells at 33rd passage was 81. Specific marker chromosome is not realized. The production of SCC antigen is detected from the cells and the amount of SCC antigen in cultured media was recorded from 1.5 to 2.0 ng per 1 x 10(4) cells for 48 hours. CEA synthesis is also confirmed immunohistochemically.