Large scale replication analysis of loci associated with lipid concentrations in a Japanese population.

BACKGROUND: Recent genome wide association studies discovered seven novel loci that influence plasma concentrations of triglycerides, high density lipoprotein (HDL) and low density lipoprotein (LDL) cholesterol in Europeans. To date, large scale replication studies using populations with known differences in genome-wide linkage disequilibrium (LD) pattern have not been undertaken. METHODS: To address this issue, we tested associations between single nucleotide polymorphisms (SNPs) within the seven novel loci and plasma lipid profiles in 21 010 Japanese individuals. RESULTS: Multiple linear regression analyses showed that the rs3812316 in MLXIPL was strongly associated with triglyceride concentrations (p approximately 3.0x10(-11), 7.1 mg/dl decrease per minor C allele) and that rs599839 in CELSR2/PSRC1/SORT1 was strongly associated with LDL cholesterol concentrations (p approximately 3.1x10(-11), 4.7 mg/dl decrease per minor G allele) in the Japanese population. SNPs near ANGPTL3, TRIB1 and GALNT2 showed evidence for associations with triglyceride concentrations (3.6x10(-6)

Vasoactive intestinal peptide in human nasal mucosa.

Vasoactive intestinal peptide (VIP), which is present with acetylcholine in parasympathetic nerve fibers, may have important regulatory functions in mucous membranes. The potential roles for VIP in human nasal mucosa were studied using an integrated approach. The VIP content of human nasal mucosa was determined to be 2.84 +/- 0.47 pmol/g wet weight (n = 8) by RIA. VIP-immunoreactive nerve fibers were found to be most concentrated in submucosal glands adjacent to serous and mucous cells. 125I-VIP binding sites were located on submucosal glands, epithelial cells, and arterioles. In short-term explant culture, VIP stimulated lactoferrin release from serous cells but did not stimulate [3H]glucosamine-labeled respiratory glycoconjugate secretion. Methacholine was more potent than VIP, and methacholine stimulated both lactoferrin and respiratory glycoconjugate release. The addition of VIP plus methacholine to explants resulted in additive increases in lactoferrin release. Based upon the autoradiographic distribution of 125I-VIP binding sites and the effects on explants, VIP derived from parasympathetic nerve fibers may function in the regulation of serous cell secretion in human nasal mucosa. VIP may also participate in the regulation of vasomotor tone.

Increased synthesis of hyaluronic acid by mouse mammary carcinoma cell variants with high metastatic potential.

Variant subpopulations of FM3A mouse mammary carcinoma cells that have increased lung-colonizing potential were obtained previously by sequentially harvesting pulmonary metastases, culturing their cells in vitro, and reestablishing the metastases in vivo. In the present study, glycosaminoglycan production by the parental and variant cells was studied after metabolic labeling of cultures by [14C]glucosamine for 24 hr. Analysis of the products indicated that the rate of incorporation of the labeled precursor into hyaluronic acid in the high-metastatic variant cells was 27 to 54 times the rate in the low-metastatic variant cells and that the increase in hyaluronic acid synthesis was not associated with an increase in the rate of synthesis of other glycosaminoglycans. Both the cell layers and media of high-metastatic variants contained a much higher proportion of radioactivity in hyaluronic acid than did the corresponding fractions of low-metastatic cell lines. The results provide a basis for further investigation of the potential role of hyaluronic acid in control of the behavior of epithelial tumor cells during metastasis.

Transformation of chicken chondrocytes by Rous sarcoma virus.

Chicken chondrocytes isolated from 11-day-old chicken vertebrate cartilage were transformed by Rous sarcoma virus ts LA24 of the Prague strain as well as by the wild-type Prague strain of Rous sarcoma virus. The morphology of chondrocytes transformed by Rous sarcoma virus ts LA 24 was dependent on the temperature, and the change was reversible. A similar but irreversible change in morphology was observed with chondrocytes transformed by wild-type virus. Hyaluronic acid production and deoxyglucose transport were markedly increased in the transformed chondrocytes. A marked increase of labeled acetate incorporation was observed with the transformed chondrocytes. In contrast to the normal chondrocytes, the labeled hyaluronic acid synthesized by the transformed chondrocytes was mostly released into the culture medium.

Characterization of a novel ceramide octasaccharide isolated from whole tissue of a fresh-water bivalve, Corbicula sandai.

A novel glycosphingolipid containing two mannose residues, provisionally named GL-1 in a previous paper (Itasaka, O. and Hori, T. (1979) J. Biochem. 85, 1469-1481), was isolated from whole tissue of a fresh-water bivalve, Corbicula sandai, and the structure of the glycolipid was determined to be: Galp4Me(beta1 more than 3)GalpNAc(beta1 more than 3)Fucp(alpha1 more than 4)GlcpNAc(beta1 more than 2)Manp(alpha1 more than 3)(Xylp(alpha1 more than 2))Manp(beta1 more than 4)Glcp(beta1 more than 1)Cer. Glycosphingolipid-1 seems to be a biosynthetic precursor of glycosphingolipid-3, because they have an identical octasaccharide structure, except for the presence of an aminoethylphosphoryl residue in glycospingolipid-3. However, in contrast to glycosphinogolipid-3, glycosphingolipid-1 has a more complex ceramide pattern. This fact suggests that glycosphingolipid-1 as isolated is not a degradation product of glycosphingolipid-3, and it can be presumed that the restricted molecular species of glycosphingolipid-1 which has the same ceramide constitution as glycosphingolipid-3 may be utilized preferentially for glycosphingolipid-3 biosynthesis.

Highly sulfated proteochondroitin sulfates synthesized in vitro by rat glomeruli.

A previous report from this laboratory (Kobayashi, S., Oguri, K., Kobayashi, K. and Okayama, M. (1983) J. Biol. Chem. 258, 12051-12057) indicated that isolated rat glomeruli synthesized three species of sulfated glycoconjugates in vitro, namely, sulfated glycoproteins, proteoheparan sulfates and proteochondroitin sulfates. In the present study, the proteochondroitin sulfates, which showed the greatest incorporation of [35S]sulfate among the three sulfated glycoconjugates, were isolated and characterized. Radiolabeled tissue proteochondroitin sulfates were clearly separated on Sepharose CL-6B into three components with partition coefficients (Kd) of 0.16, 0.22 and 0.58, and medium proteochondroitin sulfates were separated into two components with Kd values of 0.33 and 0.62. When the chondroitin sulfate chains released by alkaline borohydride treatment were analyzed by digestion with chondroitinase AC-II, chondroitinase ABC, chondro-6-sulfatase and chondro-4-sulfatase, the results showed that all the samples contained glucuronosyl-N-acetylgalactosamine (chondroitinase AC-II-susceptible sequences, 72-86%) and iduronosyl-N-acetylgalactosamine (chondroitinase ABC-susceptible sequences, 14-28%), containing 4-sulfated N-acetylgalactosamine (50-70%) and 4,6-disulfated N-acetylgalactosamine (30-50%). On two-dimensional electrophoresis on cellulose acetate, all samples gave a single spot which closely coincided with chondroitin sulfate E of squid cartilage in electrophoretic mobility. These results indicated that the chains were highly sulfated chondroitin sulfates containing glucuronic acid and iduronic acid residues.

Primary amyloidosis with familial vitreous opacities: an unusual case and family.

Peripheral neuropathy was not found even six to ten years after the onset of visual symptoms in a family with primary amyloidosis, except in the propositus at the terminal stage. The propositus had mainly ocular and CNS involvement. An ocular manifestation, the vitreous opacity, was the only involvement in the family members, in spite of the long clinical course. This family may have a different type of familial primary amyloidosis from that previously reported.

Temporal frequency deficit in patients with senile dementia of the Alzheimer type: a visual evoked potential study.

To study the temporal aspect of visual processing of senile dementia of the Alzheimer type (SDAT), we recorded the steady-state visual evoked potentials (VEPs) to LED goggle stimulation at five different temporal frequencies in 14 patients with SDAT and 14 control subjects. For comparison, we also studied 14 patients with vascular dementia (VD). All subjects were women, and their ages were closely matched. Temporal frequencies were 5, 10, 15, 20, and 30 Hz. The steady-state VEPs were Fourier analyzed, and both the amplitude and phase of the first and second harmonic responses were obtained. There was a significant difference in the amplitude across the three groups for stimulation at 15, 20, and 30 Hz, but no difference at a stimulation of 5 and 10 Hz. Multiple comparison tests revealed that patients with SDAT had a significantly smaller amplitude than normal controls. Patients with VD had lower amplitudes than normal subjects, but this was not statistically significant. In the phase data, there were also no statistically significant differences across the three groups for any of the temporal frequencies. These results indicate that a temporal frequency deficit exists in SDAT. This phenomenon may be explained by the pathology that affects the early stage of visual signal processing.

Decrease in beta-adrenergic receptors of lymphocytes in spontaneously occurring acute asthma.

To clarify changes in beta-adrenergic receptor (BAR) density in spontaneously occurring acute asthma, BAR binding studies were performed on peripheral blood lymphocytes in eight asthmatic and ten normal subjects. Spirometry also was performed. Maximum binding capacity (Bmax) of BAR in acute asthma was significantly lower by 44.2 percent compared with that in stable asthma; FEV1/FVC ratio decreased by 23.9 percent. The Bmax for acute asthma also was significantly lower than that in normal subjects. The Bmax of BAR in all subjects was significantly correlated with FEV1/FVC ratio and percent FEV1. The percentage decrease in the FEV1/FVC ratio and FEV1 from the stable to acute state for each asthmatic subject did not correlate with corresponding percentage decrease in Bmax. These data demonstrate that BAR density of lymphocytes decreases substantially in acute asthma and, simultaneously, suggest that some factors other than the BAR mechanism contribute to the airway obstruction during acute asthma.

Membrane-intercalated proteoglycan of a stroma-inducing clone from Lewis lung carcinoma binds to fibronectin via its heparan sulfate chains.

A Lewis lung carcinoma-derived low metastatic clone, P29, with a capacity to induce a fibrotic stromal response of host tissue, exhibits tumorigenesis depending on an interstitial matrix formed by the induced stromal cells. Using this clone, in the present study we isolated and characterized a membrane-intercalated proteoglycan that mediates interaction between the tumor cells and interstitial matrix. The tumor cells were cultured in the presence of [3H]glucosamine and [35S]sulfate or [35S]methionine, and hydrophobic proteoglycans were isolated by chromatography on DEAE-Sephacel and then Octyl-Sepharose CL-4B. Proteoglycans with high affinity to the octylresidue were obtained from the cell layer but not to any significant extent from the medium. By CsCl density gradient centrifugation, they were separated into bottom, middle, and top subfractions, which were shown to consist of homogeneous species with estimated M(r) values of 270,000 (named CPGIIIB), 200,000 (CPGIIIM), and 195,000 (CPGIIIT), respectively, by gel filtration on Sepharose CL-4B. These proteoglycans were intercalated into phosphatidylcholine liposomes, suggesting that they are all membrane-intercalated proteoglycans. Analyses of their glycosaminoglycans with chondroitinase ABC and heparitinase I plus II demonstrated that they all contain heparan sulfate as a major glycosaminoglycan (58-85%) and chondroitin 4-sulfate as a minor one (15-42%). Of these three proteoglycans, only CPGIIIB proteoglycan bound specifically to fibronectin-Sepharose 4B under physiological conditions. Molecular analyses of this proteoglycan by Sepharose CL-4B or SDS-PAGE before and after treatments with glycosaminoglycan degradation enzymes or trifluoromethanesulfonic acid demonstrated that CPGIIIB proteoglycan is a hybrid proteoglycan having heparan sulfate and chondroitin 4-sulfate chains on the same core protein with an M(r) of 40,000. Affinity chromatographies of the CPGIIIB proteoglycan on fibronectin-Sepharose 4B after treatments with these enzymes demonstrated that it bound to fibronectin via its heparan sulfate chains. On the basis of the above results, we propose that the CPGIIIB proteoglycan mediates the interaction between the tumor cells and interstitial matrix.

The localization of sulfated glycoconjugates synthesized by the corneal epithelium of chick embryos.

The localization of sulfated glycoconjugates in the corneal epithelium of 19-d-old chick embryo was investigated biochemically using epithelia labeled in vitro with [35S]sulfate, which exhibited autoradiographically a similar distribution of silver grains to that labeled in ovo. The radiolabeled tissues were dissociated into single cells by incubation in 0.25% trypsin containing 0.02% EDTA at 37 degrees C for 40 min. The proteoglycans and sulfated glycoproteins which were associated with the cells and those released into the dissociation medium were separated by DEAE-Sepharose CL-6B and analyzed on Sepharose CL-6B and SDS-PAGE. About 86% of the proteoglycans was released into the dissociation medium and more than 50% of the cell-associated ones was affected by trypsin. This indicates that the proteoglycans are mostly localized in an extracellular compartment. On the other hand, the extent of release of sulfated glycoproteins into the medium on dissociation of tissues was distinctly different depending upon their molecular weight (Mr): almost all of the sulfated glycoproteins of the family with Mr 48,000-70,000 (32% of the total sulfated glycoproteins) were recovered as intact molecules with the cells, whereas approximately 50% of those with Mr 70,000-150,000 (36%) and about 70% of those with Mr over 150,000 (28%) were released into the dissociation medium. These results indicate that the family with Mr 48,000-70,000 is localized intracellularly and that with Mr 70,000-150,000 in a compartment poorly affected by trypsin; in contrast to those, that with Mr more than 150,000 is localized in an extracellular compartment like the proteoglycans.

Sulfated glycoproteins synthesized by the corneal epithelium of chick embryo having the same molecular weights as cytokeratin polypeptides.

The previous study from this laboratory demonstrated that the corneal epithelium of 19-d-old chick embryo synthesizes two classes of sulfated glycoconjugates consisting of sulfated glycoproteins and proteoglycans (Yonekura, H., Oguri, K., Nakazawa, K., Shimizu, S., Nakanishi, Y., & Okayama, M. (1982) J. Biol. Chem. 257, 11166-11175). The present study demonstrated that when the sulfated glycoproteins labeled metabolically with [35S]sulfate and [3H]glucosamine were analyzed by SDS-PAGE, the 70,000 component (accounting for approximately 30% of the 35S and 35% of the 3H of the total sulfated glycoprotein) co-migrated with five major proteins with apparent molecular weights (Mrs) of 70,000, 66,000, 58,000, 51,000, and 48,000, which together accounted for about 57% of the total tissue protein. All five proteins cross-reacted with an antibody against human sole keratin, indicating that they are cytokeratin polypeptides of the corneal epithelium. Amino acid analysis demonstrated that they had high contents of glycine, serine, glutamic acid, leucine, and aspartic acid. Two-dimensional tryptic peptide maps indicated that they were all different. Analysis of radiolabeled materials released by alkaline borohydride treatment of the sulfated glycoproteins which were synthesized in the presence and absence of tunicamycin and co-purified with the five cytokeratin polypeptides, revealed that they contained both N- and O-glycosidically linked sulfated oligosaccharides. All the results obtained in the present study indicate that the five sulfated glycoproteins are similar, if not identical, to the cytokeratin polypeptides. This is consistent with the result in the accompanying paper that these sulfated glycoproteins are localized intracellularly.

Difference in blood flow volume of the common carotid artery between vascular and non-vascular dementia detected by colour duplex sonography.

Using colour duplex sonography, blood flow volume in the common carotid artery was measured in 72 demented patients and 28 normal controls. Thirty-five patients with a Hachinski's ischaemic score of 7 or above and marked ischaemic lesions on CT were assigned to the vascular dementia (VD) group. Thirty-three patients with probable Alzheimer's disease according to the NINCDS-ADRDA criteria, 2 patients with Parkinson's disease, 1 patient with spinocerebellar degeneration, and 1 patient with Pick's disease were assigned to the non-vascular dementia (nonVD) group. The sum of blood flow volume in the bilateral common carotid arteries (CCA flow) in the nonVD group and in the VD group was lower than that in the control group. The CCA flow in the VD group was lower than that in the nonVD group. Comparison of patients matched for both age and the Hasegawa's dementia rating scale also revealed lower CCA flow in the VD group than in the nonVD group. Linear discriminant function analysis showed that nearly 90% of the demented patients were correctly diagnosed as having VD or as having nonVD. These results show the usefulness of colour duplex sonography in the differential diagnosis of patients with dementia.

Possible contaminant origins of the red cosmetics decorating ancient burial sites in Japan.

Marker elements were estimated from the red cosmetics collected from different ancient burials and mine ruins in three separate districts of Japan. Element levels were displayed in reference to the relative amount to sulfur (RA/S), by which the cosmetics were divided into five types: I--a low Hg/S with a low Fe/S; II--both moderate Hg/S and Fe/S; III--a moderate Hg/S with a high Fe/S; III 2--a high Hg/S with a moderate Fe/S; IV--a high Hg/S with a high Fe/S. The cosmetics can be further characterized by referring to other contaminants such as Zn, Cu, and Mn. These combined analyses with contaminant metals were capable of characterizing the origins of the cosmetics; it is useful to compare them to each other. The cosmetics were identified as being due to several groups of contaminants from ancient mines in Japan, and also with this system analysis of the markers it is possible to identify them from neighboring countries.

[The efficacy of HFJV (high frequency jet ventilation) for ARDS (adult respiratory distress syndrome) following total arch replacement].

A 40-year-old man was admitted to the hospital with shock due to acute aortic dissection (type A of Stanford classification). The emergent total arch replacement was performed with the retrograde cerebral perfusion uneventfully. At the 3rd day after the operation, bilateral infiltrates were recognized on the frontal chest radiograph. The oxygenation was gradually decreased to less than 200 mmHg of PaO2/FiO2 in proportion with the growth of infiltrates and increase of LDH in blood. Steroid therapy was not effective. So then HFJV was introduced to improvement of hypoxygenation at the 16th day after the operation. HFJV made PaO2/FiO2 improve immediately to more than 300 mmHg and the inspiratrory pressure decrease to less than 30 mmHg. Bilateral infiltrates were diminished by HFJV. LDH in blood decreased apparently. HFJV had been used together during 15 days. The respirator was weaned at the 46th day after the operation. In this clinical course, the change of PaO2/FiO2 had the negative correlation with that of LDH in blood.

[A study of mental health in community-dwelling older persons--influence of cared elderly].

This study aimed to evaluate mental health of elderly persons living in the community and understand the various backgrounds and effects of care giving. The study used a questionnaire from similar to the one used by Matsubayashi et al in the Kahoku-cho study. The Japanese Version of General Health Questionnaire 12 (GHQ-12) was used to evaluate mental health. Responses were received from 2,799 (81.5%) of 3,432 Numura-cho residents (81.5%). After removing inadequate responses, 1,268 (36.9%) (575 men and 693 women) were analyzed. Fifty-five men (9.6%) and 69 women (10.0%) were caring for someone at home. Multiple logistic regression analysis which examined the relation between mental health and the background factors such as having someone to care for, revealed that their mental health was significantly poor compared to those having no one to care for (odds ratio (OR) = 2.01, 95% confidence interval (CI), 1.29-3.15). As for the relation between the degree of care giving and their mental health, the subjective burden was significantly poor at 72-100% (OR = 3.40, 95% CI, 1.71-6.78) and for those with three to eight years of care-giving (OR = 2.22, 95% CI, 1.00-4.90), disabled activity of daily living (almost bed ridden) of the cared for person (OR = 5.53, 95% CI, 2.23-13.7), severely demented state (OR = 5.13, 95% CI, 2.14-12.3), their mental health was significantly poor. However, being able to participate in an event (OR = 0.12, 95% CI, 0.03-0.49) and strong emotional support (OR = 0.12, 95% CI, 0.03-0.49) were negatively associated with caregivers' psychiatric distress. Having someone to care for largely affected mental health of the elderly to subjects living in the community. Services should be devised to serve the needs such as psychological care of care-givers as well as those being cared for.

[A study of depressive state and background factors in community-dwelling older persons].

In order to maintain and improve the mental health of elderly people living in the community, a cross-sectional survey was conducted to elucidate their depressive state and its background factors. Subjects were elderly persons living in the community who were able to fill in the questionnaire themselves. The study used the self-recording questionnaire sheets used in the Kahoku Longitudinal Aging Study by Matsubayashi et al and the Zung Self-rating Depression Scale (SDS). Out of 2,379 elderly persons who were able to fill in the questionnaire by themselves in the community, 2,361 (99.2%) returned the questionnaire sheets. After removing inadequate responses, analysis was possible for 1,181 (49.6%) (542 males (average age 72.3 +/- 5.5) and 639 females (average age 73.0 +/- 6.3). Degree of depressive state as evaluated by SDS was normal for 731 persons (61.9%); mild, 240 (20.3%); moderate, 181 (15.3%); and severe, 29 (2.5%). The average age became higher as the SDS became high, of being indicating the seriousness of the depressive state (p = 0.0155), with the ratio women significantly higher (p = 0.0077). Among those with severe SDS, the ratio of single persons was high (p < 0.001) as well as those who were non-drinkers (p = 0.0015), without regular habit of walking (p < 0.001), or without work (p < 0.001). The ratio of those receiving medication regularly was also significantly high (p = 0.0022). As for the relation of SDS with various of the scores, the higher the SDS score became, the scores for ADL, information-related function, functional and emotional support network, family relationship, friendship, economic condition became significantly lower (p < 0.001, respectively). In logistic regression analysis using the background factors for SDS as explanatory variables, factors such as being women (odds ratio, 1.73; 95% confidence interval, 1.10-2.72). ADL (0.80; 0.69-0.93), emotional support network (0.88; 0.81-0.96), friendship (0.98; 0.96-0.99) were significant independent contributing factors. As for the relation between SDS and subjective senses, the more serious the SDS score became, the scores for feelings of healthiness and satisfaction became significantly smaller (both p < 0.001). For prevention and amelioration of the depressive state of elderly persons living in the community, attempts should be made to improve the background factors which were clarified by the present study by efficiently utilizing health, medical and welfare services and following the future course with a positive attitude.

[Happiness and background factors in community-dwelling older persons].

In order to maintain and improve mental health of elderly people living in the community, a cross sectional survey was conducted to elucidate their happiness and background factors. The subjects were elderly persons living in the community who were able to fill in the questionnaire themselves. The study employed the self-recording questionnaire forms used in Kahoku Longitudinal Aging Study by Matsubayashi et al. Happiness was assessed using a visual analogue scale. Out of 2,379 elderly persons who were able to fill in the questionnaire by themselves in 2,361 (99.2%) returned the questionnaire sheets. After removing inadequate responses, analysis was possible for 1,873 (78.7%) (860 men (average age 72.7 years) and 1,013 women (average age 72.8 years). Among those with greater happiness, the ratio of those living with others (p = 0.0051) was high as well as those with spouses (p = 0.0240), without a history of hypertension (p = 0.0096) and apoplexy (p = 0.0039), not receiving medication regularly (p = 0.0039), with regular habit of walking (p < 0.001), or with work (p < 0.001). As for the relationship between happiness and various scores, the higher the happiness scale became, the scores for ADL, information-related function, functional and emotional support network, healthy condition, appetite condition, sleep condition, memory condition, family relationships, friendship, economic condition became significantly higher (p < 0.001, respectively). On multiple regression analysis using the background factors for happiness as explanatory variables, factors such as functional support network (p < 0.001), emotional support network (p = 0.0254), healthy condition (p < 0.001), good memory condition (p = 0.0027), friendship (p < 0.001), good economic condition (p < 0.001) were significant independent contributing factors. As for the relation between SDS and happiness, the more serious the SDS score (higher score) became, the scores for the feeling of happiness became significantly smaller (p < 0.001). For amelioration of the happiness of elderly persons living in the community, attempts should be made to improve the background factors clarified by the present study by efficiently utilizing health, medical and welfare services.

Acyclovir-resistant herpes zoster encephalitis successfully treated with vidarabine: a case report.

A 78-year-old man developed herpes zoster virus (HZV) encephalitis. Initially, treatment with aciclovir (750 mg per day) improved CSF cell count and protein level. During the treatment, however, encephalitis in the patient deteriorated in spite of the treatment with aciclovir, suggesting that HZV in the patient had become resistant to aciclovir. Subsequent treatment with vidarabine (600 mg per day, for 15 days) resulted in dramatic improvement in CSF pleocytosis. About two months after the discontinuation of vidarabine, the CSF cell count was normal. The patient became alert gradually, but his amnestic syndrome remained unchanged. Vidarabine may be recommended in the treatment of HZV encephalitis when aciclovir is not effective.