Aberrant expression of microRNAs and the miR-1/MET pathway in canine hepatocellular carcinoma.

Canine hepatocellular carcinoma (HCC) is the most common primary hepatic tumour in dogs. MicroRNA (miRNA) dysregulation has been reported in human HCC and shown to have diagnostic and prognostic value; however, there are no data on miRNA expression in canine HCC. The aim of the present study was to investigate differentially expressed miRNAs in canine HCC. Analysis of miRNA expression in canine HCC tissues and cell lines by quantitative reverse transcription PCR showed that miR-1, miR-122, let-7a, and let-7g were downregulated, whereas miR-10b and miR-21 were upregulated in canine HCC. MET is one of the target genes of miR-1. MET was upregulated in canine HCC at the gene and protein levels, and a significant correlation between the concomitant downregulation of miR-1 and upregulation of MET was observed. Fast/intermediate-proliferating canine HCC cell lines had higher MET gene and protein expression levels than the slow-proliferating cell line. These findings suggest that miRNAs are differentially expressed in canine HCC, and that the miR-1/MET pathway may be associated with canine HCC cell proliferation.

The association between neutropenia and prognosis in stage III colorectal cancer patients receiving adjuvant chemotherapy.

Neutropenia during chemotherapy has been reported to be a predictor of better survival in patients with several types of cancer, although there are no reports on stage III colorectal cancer (CRC). The purpose of this study was to examine the association between neutropenia and prognosis in stage III CRC patients receiving adjuvant chemotherapy consisting of oral uracil and tegafur (UFT) plus leucovorin (LV). We retrospectively analysed 123 patients with stage III CRC who received UFT/LV as adjuvant chemotherapy. The end-point was disease-free survival (DFS). Survival curves of the two categories (neutropenia absent vs. present) were estimated using the Kaplan-Meier method and compared by the log-rank test. We estimated the hazard ratio (HR) for DFS according to neutropenia after adjustment for covariates by multivariate analyses using Cox's regression analysis. A total of 33 (26.8%) patients experienced neutropenia. Patients without neutropenia showed a significantly lower DFS than those with neutropenia (3-year DFS 57.3% vs. 81.2%, P = 0.0213). By multivariate analysis, neutropenia and histological type were independent prognostic factors, with HR of 0.410 (neutropenia absent vs. present, P = 0.045) and 4.793 (well to moderately differentiated vs. poorly differentiated, P = 0.004) respectively. We demonstrated that neutropenia occurring during adjuvant chemotherapy consisting of UFT/LV may be a prognostic factor of recurrence in stage III CRC patients.

OARE-1, a Ty1-copia retrotransposon in oat activated by abiotic and biotic stresses.

Transcriptionally active Ty1-copia LTR-retrotransposons were found in oat using RT-PCR for amplifying the reverse transcriptase domain. Sequence analysis of the RT-PCR clones suggested that oat LTR-retrotransposons consist of at least seven groups, which were tentatively designated as Oatrt1 to Oatrt7. A full length copy of Oatrt1 was isolated from an oat genomic library, and was designated OARE-1. OARE-1 was 8,665 bp long and a member of the BARE-1 subgroup. The oat genome carried it in multiple copies (at least 10,000 copies / a hexaploid genome). The expression of OARE-1 was intensively induced by wounding, UV light, jasmonic acid and salicylic acid, and its pattern was very similar to that of the PAL (phenylalanin ammonia lyase) gene. Furthermore, OARE-1 was highly activated by infection with an incompatible race of the crown rust fungus, Puccinia coronata. These results suggest that OARE-1 is highly sensitive to various abiotic and biotic stimuli leading to plant defense responses.

Possible risk factors of carotid artery atherosclerosis in the Japanese population: a primary prevention study in non-diabetic subjects.

OBJECTIVE: Hyperinsulinemia has been associated with the risk of coronary heart disease, stroke, and renal disease in nondiabetic subjects. However, direct evidence that hyperinsulinemia per se is directly associated with atherosclerosis has been conflicting. The present study was designed to investigate the cross-sectional association of carotid artery atherosclerosis with insulin, independent of well-known cardiovascular risk factors, in nondiabetic subjects. METHODS AND SUBJECTS: Between 1996 and 1997, 1,335 subjects (620 men and 715 women) were recruited from one Japanese community, interviewed, and examined. Clinical measurements in the study included intimal-medial thickness (IMT) of the carotid artery, fasting plasma insulin, serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), hemoglobin type HbA1c, systolic blood pressure (SBP), diastolic blood pressure (DBP), and body mass index (BMI). We divided the subjects of both genders into three subgroups according to age (40-49 years of age; 50-59; and 60-69). RESULTS: Using simple regression analysis, we found that IMT was significantly correlated with at least one of TC, LDL-C, HbA1c, SBP, DBP, and BMI in each subgroup. The results of multivariate analysis showed that IMT was independently correlated with TC, HDL-C, LDL-C, SBP and BMI in males and with TC, TG, HDL-C, LDL-C, HbA1c, SBP, DBP, and BMI in females. Insulin levels showed no correlation with IMT in either males or females. CONCLUSION: Fasting hyperinsulinemia does not appear to be correlated with carotid artery atherosclerosis based on the present cross-sectional results.

Patients' attitude toward consultations by a physician without a white coat in Japan.

OBJECT: To know how Japanese patients perceive their physicians without a white coat during consultations. SUBJECTS AND METHODS: The patients who visited a university clinic were divided into two groups: those seen by a physician in a white coat (the white-coat group) and those seen by a physician in private clothes (the private-clothes group). Questionnaires were distributed to the patients, which asked the tension and satisfaction of consultations as well as their preference for physician's attire. The answers of the white-coat group were compared with those of the private-clothes group. RESULTS: The percentage of new patients who felt tense during consultations was greater in the white-coat group (42%) than in the private-clothes group (33%). Seventy-one percent of the patients in the white-coat group preferred physicians in a white coat whereas only 39% preferred so in the private-clothes group (p<0.0001). However, the degree of patients' satisfaction for the consultation showed no statistical difference between the groups. Sixty-nine percent of the patients older than or equal to 70 years preferred a white coat while 52 percent of the patients younger than 70 years preferred so (p=0.002). CONCLUSION: Physician's white coats did not influence the satisfaction with the consultations for most Japanese patients in a university clinic, although elderly patients as well as those seen by a physician in a white coat tended to prefer the white coat to the private clothes. Furthermore, practice without a white coat might reduce patients' tension during their first consultation.

Three-dimensional large-screen display with reflection-mode spatial light modulators and a single-projection optical system: analysis of a retardation-modulation method.

A stereoscopic projector with polarized glasses is proposed that consists of spatial light modulators (SLM's) that control the retardation of projected light, a polarization beam splitter (PBS), and single-projection optics. This display's features include a three-dimensional (3-D) image display with a single projector and half the size and the power consumption of a conventional 3-D projector. Analysis shows that the cross talk and the extinction ratio of this system depend strongly on the polarized light-separation characteristics of the PBS, the light output, and the extinction ratio of the SLM's. A double-PBS method that drastically improves 3-D image quality is also discussed.

Effects of interferon-alpha, beta, and gamma on the function of differentiated leukemic HL-60 cells induced by 1,25-dihydroxyvitamin D3.

The differentiation of HL-60, a human leukemic cell line, into monocyte-like cells (D3-HL-60 cells) is induced by 1,25-dihydroxyvitamin D3 (D3). We examined the effects of interferon (IFN) treatment of D3-HL-60 cells on the expression of cell surface antigens, the phagocytic activity for fluorescent beads, production of oxygen radicals, and intracellular growth of Legionella pneumophila. Activation of D3-HL-60 cells with IFN-gamma, Beta, and alpha for 24 h significantly increased expression of CD16, CD36, CD71, and HLA-DR antigens. IFN-gamma markedly enhanced the phagocytic activity of beads in D3-HL-60 cells. There was no significant difference in phagocytic activity between cells exposed to IFN-alpha or beta and untreated D3-HL-60 cells. IFN-alpha, beta, and gamma enhanced production of oxygen radicals, including superoxide, by D3-HL-60 cells. Superoxide production was enhanced to the greatest degree by IFN-gamma, followed by IFN-beta and then IFN-gamma. Intracellular growth of L. pneumophila in D3-HL-60 cells was inhibited by interferons (IFN-gamma > beta > gamma). Similar results were obtained in human mononuclear cells. These data indicate that interferons can act as biologic response modifiers not only in human mononuclear cells but also in differentiated leukemic cells. Our results may have implications for the development of differentiation therapy for treatment of leukemia.

Granule-laden perivascular cells observed in rabbits with experimental cerebral atherosclerosis.

Granule-laden perivascular cells distributed around the fine vessels in brain of hypertensive rabbits fed a cholesterol diet were examined by electron microscopy. Perivenule granule-laden cells contained secondary lysosomes, residual bodies, Golgi vesicles, fusion vesicles, and vacuoles. In periarteriole granule-laden cells, secondary lysosomes and vacuoles were not prominent. Findings indicated that the granule-laden cell may be a histiocyte that appears during the vascular reaction to hypertension and hypercholesterolemia. This vascular reaction may occur more strongly in the veins than the arteries.

Urine C-peptide and atherogenic risk factors in diabetes mellitus: relevance to "syndrome X".

The relation between the C-peptide concentration in twenty-four-hour urine specimens and atherogenic risk factors was investigated in 38 patients with noninsulin-dependent diabetes mellitus in an attempt to determine the significance of urine C-peptide in diagnosing "syndrome X," which is characterized by insulin resistance. Weak positive correlations between twenty-four-hour urine C-peptide concentration and body mass index, systolic blood pressure (BP), diastolic BP, serum total cholesterol, and serum triglyceride were detected. A weak negative correlation was also apparent between urine C-peptide and serum high-density lipoprotein (HDL). The body mass index and serum triglyceride of patients with urine C-peptide excretion of > 100 micrograms/day were significantly higher than those in patients with normal urine C-peptide excretion (< 100 micrograms/day) (P < 0.01 and P < 0.02, respectively). Systolic BP, diastolic BP, serum total cholesterol, and serum HDL did not differ significantly between the two groups of patients. Results indicate that twenty-four-hour urine C-peptide concentration is of significance in determining whether a patient has a tendency to insulin resistance but has only limited value as a quantitative measure of endogenous insulin secretion.

Subclavian steal phenomenon associated with stress polycythemia. A case report.

A case of stress polycythemia without symptoms of cerebral ischemia is presented. The patient demonstrated a blood flow reversal through the right vertebral artery that was caused by a complete obstruction of the right subclavian artery at its origin. This obstruction may have been due to thrombosis associated with stress polycythemia. This is a rare example of a case in which thrombotic complications of stress polycythemia occurred in a larger caliber vessel such as the subclavian artery.

Effects of a new calcium channel blocker, MPC-1304, on blood pressure, serum lipoproteins and serum carbohydrate metabolism in patients with essential hypertension.

The effects of MPC-1304, a new calcium channel blocker, on blood pressure, serum lipoproteins, and carbohydrate metabolism were compared with those of atenolol in a group of patients with mild to moderate essential hypertension. Systolic and diastolic pressures were significantly decreased by both MPC-1304 and atenolol administration. Serum levels of apolipoproteins A-I and A-II were significantly increased after 8-12 weeks of MPC-1304 treatment, but were unchanged during a similar period of atenolol treatment. Neither drug induced any significant change in other lipoprotein parameters, fasting blood sugar, immunoreactive insulin, C-peptide or HbA1c. No serious side-effects or abnormal laboratory values were observed during the course of administration of either drug. These findings indicated that MPC-1304 is as efficacious as an antihypertensive drug and is without adverse effect on lipoprotein or carbohydrate metabolism.

The mechanism of interaction of sodium dodecyl sulfate with elastic fibers.

Sodium dodecyl sulfate (SDS), an anionic hydrophobic ligand, is known to alter the mechanical properties of elastic fibers. In order to analyze the mechanism of the alteration, two forms of fibrous elastins, "solid" and "powder" elastins, which consisted of fascicular elastic fibers and single or oligomeric elastic fibers, respectively, were prepared from bovine aorta, and the interactions of SDS with these elastins in the presence and absence of 0.15 M NaCl were studied. The solid elastin was able to retain 1.2- to 1.4-fold larger amounts of SDS than the powder elastin under both conditions, and both elastins retained 1.2-fold or larger amounts of SDS in the presence of NaCl than in its absence. Whereas both elastins released the retained SDS gradually on repeated washing with an SDS-free buffer, the release rates from the solid elastin, especially the rate in the presence of NaCl, were much smaller than those from the powder elastin, and the solid elastin retained approximately 40% of the bound SDS under conditions where the powder elastin lost most of its SDS. The SDS-binding capacities of both elastins were significantly lower than those of soluble kappa-elastin and serum albumin, which bound SDS homogeneously on the polypeptide chains. When the washed SDS-bound solid elastin was incubated with methylene blue and examined under a microscope, most of the methylene blue-SDS complex was located at the interfiber spaces of the elastic fibers. These results suggest that SDS alters the mechanical properties of elastic fibers by binding to the interfiber spaces and surfaces of the fibers rather than by binding to the internal polypeptide chains.

Gadolinium-enhanced magnetic resonance imaging of meningeal carcinomatosis in colon cancer.

Meningeal carcinomatosis has been infrequently reported in colorectal carcinoma. Recently the value of gadolinium-enhanced magnetic resonance imaging (MRI) in the examination of this disease has been described. We report a 55-year-old man with meningeal carcinomatosis from a known colon cancer. In this case, a gadolinium-enhanced MRI of the brain showed enhancing dura and nodular lesions which were not detected by enhanced computed tomography. The gadolinium-enhanced MRI may serve as an indicator of leptomeningeal involvement and may be a useful test in addition to the evaluation of cerebrospinal fluid cytology and chemistry.

Cerebral infarction in a young adult associated with protein C deficiency. A case report.

Protein C deficiency is a cause of thromboembolic disease. Venous thrombosis is the most common clinical manifestation. Arterial thrombosis is unusual and involvement of the intracranial arteries is especially rare. Herein the authors describe a case of cerebral [correction of cerebellar] infarction associated with protein C deficiency and review the relevant medical literature. A thirty-year-old man was hospitalized because of dysarthria, right limb ataxia, and a gait disturbance. Cranial computed tomography disclosed an infarction in the right cerebellar hemisphere and brachium pontis. Three months earlier the patient had had a transient ischemic attack with truncal ataxia and gait disturbances. On admission, the protein C antigen was 57% and protein C activity was 45%. Investigation of family members revealed protein C deficiency in an uncle. Literature review of stroke cases associated with protein C deficiency revealed that most had had a previous vascular event and/or a positive family history or had used oral contraceptives chronically. Protein C deficiency should be considered in young stroke patients with a positive family history of vaso-occlusive disease, previous ischemic events, or chronic oral contraceptive use.

Biochemical characterization of the fibres of collagen and elastin in the aortic wall of SHRSP.

1. Vessel wall fragments consisting of collagen, elastin and other insoluble proteins were prepared from the aortas of 6 month old WKY and SHRSP and dead, elderly humans. 2. Prolonged incubations of these fragments with pepsin below or at 30 degrees C resulted in different amounts of insoluble materials containing similar or larger proportions of collagen and other insoluble proteins than the respective vessel fragments. The amounts of the pepsin-insoluble materials obtained from SHRSP were larger than those from WKY but were much smaller than those from elderly humans. 3. The elastins isolated from the vessel fragments were solubilized by pepsin much more effectively than the respective vessel fragments. 4. The pepsin-insoluble materials from WKY were composed of thin mesh-shaped materials, while these materials from both rats did not contain a significant number of distinctive fibrils of collagen, the materials from elderly humans did contain numerous distinctive fibrils of collagen. 5. Large fractions of both the collagen and other proteins in the pepsin-insoluble materials were solubilized by incubation with a crude bacterial collagenase below 30 degrees C or by incubation with pepsin above 40 degrees C where the triple-helical regions of the collagens were unfolded. 6. These results appear to indicate that the aortic wall of SHRSP contains larger amounts of some insoluble components that immobilize the collagen fibrils than that of WKY, but the aortic walls of elderly humans contain much larger amounts of these components than that of SHRSP.

Comparative effects of monatepil, a novel calcium antagonist with alpha 1-adrenergic-blocking activity, and nitrendipine on lipoprotein and carbohydrate metabolism in patients with hypertension.

The effects of monatepil, a new calcium antagonist with alpha 1-blocking activity, and nitrendipine on lipoprotein and carbohydrate metabolism in 86 patients with mild-to-moderate hypertension were examined in a randomized, open-label, multicenter (32 hospitals) study. Thirty-nine patients treated with monatepil and 33 patients treated with nitrendipine completed the 12-week study. Monatepil and nitrendipine each significantly decreased both systolic and diastolic blood pressure. Changes in heart rate were not seen in either group. Monatepil administration significantly decreased total cholesterol, low density lipoprotein (LDL) cholesterol, the LDL cholesterol to high density lipoprotein (HDL) cholesterol ratio, apolipoprotein (Apo) B levels, and HbA1c levels, whereas no changes in these measurements were observed in nitrendipine-treated patients. Monatepil also significantly decreased lipoprotein(a) levels, but there were no significant changes in HDL cholesterol, Apo-AI, or Apo-E levels. After nitrendipine treatment, the C peptide concentration decreased significantly, although no significant changes were observed in fasting blood glucose or immunoreactive insulin levels. On the basis of these results, it can be concluded that monatepil belongs to a new class of antihypertensive calcium antagonist with favorable carbohydrate metabolism and lipid-lowering activity, although the clinical importance of these findings has not been established.

[An elderly patient with acute idiopathic thrombocytopenic purpura (ITP)].

An elderly patient with acute-type ITP is reported. An 89-year-old man was admitted to the Department of Physiatrics, Ureshino-Onsen Hospital on July 19, 1991 and commenced rehabilitation treatment. Laboratory data on January 28, 1992 showed decreased platelets (2.8 x 10(4)/mm3). According to this data, the patient was transferred to the Department of Internal Medicine, where laboratory data showed positive occult blood in urine and stool, prolonged bleeding time (over 10 min.) and positive PAIgG (630 ng/10(7) cells). Bone marrow showed an ITP pattern (megakaryocytes 55/mm3). The patient received steroidal pulse therapy (methylprednisolone 1,500 mg/day for 3 days). Subsequently, serum platelet count increased gradually and reached 10 x 10(5)/mm3 in May 1992. We reviewed the case reports of ITP published in Japanese, and conclude that this case was the oldest reported patient with acute-type ITP in Japan.

[The influence of serum separators on biochemical values in experimental animals].

The influence of three serum separators, A, B and C, on biochemical values was examined. With A or B, changes in the values of ChE, LAP, UN, K, P, Cl and Ca in rat serum; ChE, UN, Cl and Ca in dog serum; and K and P in monkey serum took place over a period of 20 min after blood collection. Therefore, the biochemical values of whole blood were considered to be stable after 20 min. Thus, biochemical tests were conducted on serum from the three serum separators after allowing the blood to stand 20 min. The values obtained for each separator were not markedly different from those of the control. These results suggest that a serum separator is useful for separation of serum of experimental animals under the proper conditions.

Heterogeneity of rabbit aortic endothelial cells, with special reference to phagocytosis.

Heterogeneity of aortic endothelial cells with regard to phagocytotic ability was examined by injecting India ink into normal rabbits. Light and electron microscopic analyses revealed that particles of India ink were phagocytosed in the endothelial cells, which in turn were localized at the distal side of the orifice of aortic branches, especially those of brachiocephalic, left clavicular, and dorsal intercostal arteries. No remarkable differences were found ultrastructurally between phagocytosing and nonphagocytosing endothelial cells. Ingested India ink particles were present within phagosomes of the endothelial cells for several hours after injection; the particles eventually accumulated in the subendothelial space twenty-four hours after injection. These results indicate that an active transport system of large molecules via the phagocytotic processes is present in endothelial cells located at the distal sides of the orifice of aortic branches. These regions are known to develop initial atherosclerotic lesions in hypercholesterolemic animals. Thus, a possible correlation between phagocytotic ability of endothelial cells and development of atherosclerosis is suggested.