A Novel IFITM5 Variant Associated with Phenotype of Osteoporosis with Calvarial Doughnut Lesions: A Case Report.

Osteogenesis imperfecta (OI) and other decreased bone density disorders comprise a heterogeneous group of heritable diseases with skeletal fragility. Recently, it was discovered that mutations in SGMS2, encoding sphingomyelin synthetase 2, result in aberrant sphingomyelin metabolism and lead to a novel form of OI termed osteoporosis with calvarial doughnut lesions (OP-CDL) with moderate to severe skeletal fragility and variable cranial hyperostotic lesions. This study describes a Japanese family with the skeletal phenotype of OP-CDL. The affected individuals have moderately severe, childhood-onset skeletal fragility with multiple long-bone fractures, scoliosis and bone deformities. In addition, they exhibit multiple CDLs or calvarial bumps with central radiolucency and peripheral radiopacity. However, SGMS2 sequencing was normal. Instead, whole-exome sequencing identified a novel IFITM5 missense mutation c.143A>G (p.N48S) (classified as a VUS by ACMG). IFITM5 encodes an osteoblast-restricted protein BRIL and a recurrent c.-14C>T mutation in its 5' UTR region results in OI type V, a distinctive subtype of OI associated with hyperplastic callus formation and ossification of the interosseous membranes. The patients described here have a phenotype clearly different from OI type V and with hyperostotic cranial lesions, feature previously unreported in association with IFITM5. Our findings expand the genetic spectrum of OP-CDL, indicate diverse phenotypic consequences of pathogenic IFITM5 variants, and imply an important role for BRIL in cranial skeletogenesis.

Relationship between the combination of polyunsaturated fatty acids intake and psychological distress during pregnancy: The Tohoku Medical Megabank Project Birth and Three-Generation Cohort Study.

No studies have examined the association of the combination of n-3 polyunsaturated fatty acids (PUFAs) and n-6 PUFAs intake with psychological distress during pregnancy. To examine these associations, we divided Japanese pregnant women into 25 groups based on combining quintiles of n-3 PUFAs intake and quintiles of n-6 PUFAs intake. We conducted multivariable logistic regression analyses to assess the risk of psychological distress during pregnancy (Kessler Psychological Distress Scale ≥ 5 or 13). Compared to the third quintile of both n-3 PUFAs and n-6 PUFAs intake, the groups with unbalanced intake, high intake of both, and low intake of both were associated with a higher risk of both Kessler Psychological Distress Scale ≥ 5 and 13 in early and mid-pregnancy. Further research is needed to identify the precise combination of n-3 PUFAs and n-6 PUFAs intake associated with the lowest psychological distress during pregnancy.

A resonating broken symmetry configuration interaction approach for double-exchange magnetic systems.

The correct description for ion-radical systems has recently attracted much attention from density functional theory (DFT) researchers. Although several hybridization schemes using exact (Hartree-Fock) exchange and DFT exchange-correlation functionals have been proposed, it has been reported that such treatments do not work for the description of ion-radical systems. In this study we show that combining the exact exchange term in the Kohn-Sham DFT (or the Hartree-Fock equation) with the following resonating configuration interaction method is effective for the description of double-exchange type molecular magnetic interactions. The results are analyzed in relation to the 'many-electron self-interaction' concept that was recently proposed by DFT researchers.

Copy number loss in the region of the ASPN gene in patients with acetabular dysplasia: ASPN CNV in acetabular dysplasia.

OBJECTIVES: We have previously investigated an association between the genome copy number variation (CNV) and acetabular dysplasia (AD). Hip osteoarthritis is associated with a genetic polymorphism in the aspartic acid repeat in the N-terminal region of the asporin (ASPN) gene; therefore, the present study aimed to investigate whether the CNV of ASPN is involved in the pathogenesis of AD. METHODS: Acetabular coverage of all subjects was evaluated using radiological findings (Sharp angle, centre-edge (CE) angle, acetabular roof obliquity (ARO) angle, and minimum joint space width). Genomic DNA was extracted from peripheral blood leukocytes. Agilent's region-targeted high-density oligonucleotide tiling microarray was used to analyse 64 female AD patients and 32 female control subjects. All statistical analyses were performed using EZR software (Fisher's exact probability test, Pearson's correlation test, and Student's t-test). RESULTS: CNV analysis of the ASPN gene revealed a copy number loss in significantly more AD patients (9/64) than control subjects (0/32; p = 0.0212). This loss occurred within a 60 kb region on 9q22.31, which harbours the gene for ASPN. The mean radiological parameters of these AD patients were significantly worse than those of the other subjects (Sharp angle, p = 0.0056; CE angle, p = 0.0076; ARO angle, p = 0.0065), and all nine patients required operative therapy such as total hip arthroplasty or pelvic osteotomy. Moreover, six of these nine patients had a history of operative or conservative therapy for developmental dysplasia of the hip. CONCLUSIONS: Copy number loss within the region harbouring the ASPN gene on 9q22.31 is associated with severe AD. A copy number loss in the ASPN gene region may play a role in the aetiology of severe AD.Cite this article: T. Sekimoto, M. Ishii, M. Emi, S. Kurogi, T. Funamoto, Y. Yonezawa, T. Tajima, T. Sakamoto, H. Hamada, E. Chosa. Copy number loss in the region of the ASPN gene in patients with acetabular dysplasia: ASPN CNV in acetabular dysplasia. Bone Joint Res 2017;6:439-445. DOI: 10.1302/2046-3758.67.BJR-2016-0094.R1.

NaCl-activated nucleoside diphosphate kinase from extremely halophilic archaeon, Halobacterium salinarum, maintains native conformation without salt.

Enzymes from extremely halophilic archaea are readily denatured in the absence of a high salt concentration. However, we have observed here that a nucleoside diphosphate kinase prepared from Halobacterium salinarum was active and stable in the absence of salt, though it has the amino acid composition characteristic of halophilic enzymes. Recombinant nucleoside diphosphate kinase expressed in Escherichia coli requires salt for activation in vitro, but once it acquires the proper folding, it no longer requires the presence of salts for its activity and stability.

Changes in the migratory ability of human lung and skin fibroblasts during in vitro aging and in vivo cellular senescence.

The migration of human lung and skin fibroblasts was determined during in vitro aging and in vivo cellular senescence by measuring their migration from the edge of a denuded area of a monolayer. The migration of human fetal lung fibroblasts (TIG-1 and TIG-3) decreased only very slightly with increasing passage, whereas the migration of human fetal skin fibroblasts (TIG-3S) declined gradually: the difference in cell migratory ability between early and late passages was significant (P less than 0.05). The migratory patterns of skin fibroblasts from adult and elderly donors were also similar to that of fetal skin fibroblasts. Next, the migratory abilities of fibroblast lines from adult and elderly donor groups were compared, using relatively early passaged cells. The migratory ability of the elderly-donor skin fibroblast lines was significantly lower (P less than 0.05) than that of the adult-donor skin fibroblast lines. Addition of suramin and monensin suppressed the migration of fibroblasts from fetal, adult and elderly donors, which implies that fibroblast migration is regulated by growth factors and matrix substances. The relationships between the age-dependent decline of migratory ability, growth factors and the extracellular matrix are discussed.

Age-related changes in the mitogenic activity of heparin-binding growth factors in rat sera.

Sera from rats of either sex and different ages were examined for their ability to stimulate DNA synthesis in BALB/c 3T3 cells. The activity levels of sera from male and female rats were almost the same, with age-related changes in activity also being quite similar. Activity was considerably higher in infant rats (1-month-old), but then, at a young age (6-7 months), decreased drastically for male rats, but not significantly for female rats. It increased again in middle-aged rats (12-13 months old) and was maintained at the same level toward old age (24-26 months old) for both sexes. In order to determine what kinds of growth factors were responsible for these changes, we carried out heparin affinity chromatography on the sera of male rats. Four peaks were obtained for all sera, with individual peaks exhibiting specific age-related changes in activity. Among them a peak which was eluted at 1.1 M NaCl had very high activity. It showed a similar age-related change to that of the whole sera, except for a significant increase at old age, and the factor(s) included in the peak was found to be derived from platelets. These results suggested that the factor(s) in the peak was responsible for maintaining serum mitogenic activity at an old age. The experiments undertaken to characterize this factor suggested that it is a novel one.

Interferon-beta, an autocrine cytokine, suppresses human fetal skin fibroblast migration into a denuded area in a cell monolayer but is not involved in the age-related decline of cell migration.

The migration of human skin fibroblasts into a denuded area in a cell monolayer declined during in vitro and in vivo aging. We carried out a study to determine whether this age-related decline in cell migration was mediated by the autocrine cytokine interferon-beta (IFN-beta), which has been reported to suppress the proliferation, chemotaxis and collagen synthesis of human fibroblasts. Actually, IFN-beta specifically suppressed the migration of TIG-3S human fetal skin fibroblasts into a denuded area in a cell monolayer, as shown by the dose response experiments of IFN-beta and neutralizing anti-IFN-beta antibody. IFN-beta also inhibited their collagen synthesis but the addition of type I collagen could not reverse IFN-beta-induced inhibition of cell migration. Double strand RNA, which has been generally known to induce IFN-beta in human skin fibroblasts, suppressed the migration of TIG-3S cells. Next, a study was done to determine whether IFN-beta and double strand RNA suppressed the migration of TIG-3S cells in late passages as well as early passages, or whether neutralizing anti-IFN-beta antibody stimulated the migration of TIG-3S cells in late and middle passages. IFN-beta and double strand RNA suppressed the migration of TIG-3S cells in middle (PD45) and late (PD55) passages as well as in early passages (PD23-28). Neutralizing anti-IFN-beta antibodies could not reverse the low migratory activity of middle and late passage cells to the high migratory activity of early passage cells. These results indicated that the autocrine cytokine IFN-beta did not seem to be involved in the age-dependent decline of fibroblast migration.

Effects of serum collected from rats of different ages on in vitro cell proliferation.

It has been reported by Carrel and his co-workers that serum from old hens inhibits cell growth in culture. However, as we had previously demonstrated contradictory results using serum from old rabbits, we examined whether serum from old rats would also show strong induction of cell proliferation. Sera from young and adult rats of either sex strongly stimulated the growth of rat fetal skin fibroblasts and human fetal lung fibroblasts (TIG-1). Sera of old female and male rats (24-29 months old) produced much greater fluctuations in growth-stimulatory activity than sera from young animals. Most samples of serum from old rats stimulated the growth of TIG-1 cells, as did fetal bovine serum and samples from younger rats, even when a higher concentration of serum (up to 50%) was used. On the other hand, a small proportion of samples repressed the growth of the cells. A study on the effects of serial mixtures of both different types of serum samples from old rats on cell growth suggested that this minor proportion of serum samples contain a large amount of inhibitory factor(s). The cell growth-stimulatory activity of serum did not correlate with the total protein and albumin concentrations, albumin/globulin ratio, and the levels of lipid peroxide in the sample. These results therefore seemed to imply that serum induced a striking increase in the heterogeneity of cell growth stimulatory activity with age, although most samples of serum from old rats of either sex stimulated cell proliferation as effectively as samples from younger rats. The biological significance of the small proportion of serum samples from old rats which do inhibit cell proliferation was discussed.

Effects of serum from human subjects of different ages on migration in vitro of human fibroblasts.

A study was carried out to determine whether human serum from older subjects inhibited cell migration. Sera of both sexes from subjects in their 60s (60-64 years) tended to be more inhibitory (8-14%) to the migration of human fetal lung fibroblasts, TIG-1, than serum from subjects in their 20s (20-29 years). In the case of females, the effects of serum on cell migration were significantly (P less than 0.05) different between the younger and older groups. Next, cell migration-stimulatory activity of serum was measured using human skin fibroblasts from young adult (age 21) and elderly (age 65) donors. The results were similar to those obtained with TIG-1 cells. However, the cell migration-stimulatory activity of serum was not significantly different between the two age groups. A study on the effects of concentration of human serum on the migration of TIG-1 cells showed that cell migration-stimulatory activity of serum declined linearly with increasing concentrations of sera from subjects in their teens (16-19 years) and 50s (50-59 years), and was the same between the two age groups. These results imply that substance(s) inhibitory to cell migration may not have accumulated in serum during the ageing process in humans, although human serum contained substance(s) inhibitory to cell migration.

Age-related changes in serotonin content and its release reaction of rat platelets.

The serotonin content of platelets, serum and plasma from rats of various ages was examined. In male rats, platelet serotonin content, which was about 0.65 nmol/10(8) platelets at young age (6-7 months), increased slightly at middle age (12-14 months) but decreased markedly at old age (25-26 months). Significant difference (P less than 0.05) was observed between young and old rats, and between middle-aged and old rats. In female rats, on the other hand, no age-related change in the platelet serotonin content was found. In both sexes, the serotonin content of rat sera changed with age in the same pattern as that of the platelets. No plasma serotonin was detected in rats of either sex and at any ages examined. Serotonin release from rat platelets was also studied using collagen and thrombin as stimulants. In males, the responsiveness of platelets to these two stimulants showed almost the same age-dependent changes. It was lower in middle-aged rats than in young rats but increased greatly in old rats. Significant difference (P less than 0.05) was observed between middle-aged and old rats. In females, collagen and thrombin had the opposite effect on the sensitivity of the platelets as age increased. The amount of serotonin released in response to collagen was low until middle age but increased markedly at old age, while the content of serotonin released by thrombin remained high until middle age and decreased greatly at old age. These results imply that age-related changes in the serotonin release reaction in rat platelets differed according to the stimulants used.

Effects of aging and dietary restriction on activity of monkey serum in promoting fibroblast migration.

In order to determine whether serum modified cellular aging in vivo, we previously studied the effects of serum from various mammals of different ages on cell functions such as proliferation and migration, and reported that cell migration was more greatly inhibited by serum from old donors than cell proliferation [1]. Moreover, since dietary restriction has been reported to extend lifespan and slow the aging rate of some animals [2], we wondered whether sera from dietary restricted and control monkeys of various ages might exhibit reduced aging effects on cell migration. When serum from young adult (3-5 years old) monkeys was added to plain medium, the migration of human fetal skin fibroblasts was very strongly inhibited compared to FBS. Surprisingly, sera from adult (6-11 years old) and old (more than 18 years old) monkeys caused significantly less migration-inhibitory activity than serum from young adult monkeys although sera from adult and old monkeys were much more inhibitory to cell migration than FBS. Dietary restriction only caused marginal effects on serum migration-promoting activity in a few monkey groups. The inhibition of cell migration caused by monkey serum was not brought about by cytotoxic effects since monkey serum stimulated cell proliferation as well as fetal bovine serum. These results indicate that the effects of aging on monkey serum migration-promoting activity are much more pronounced than those of dietary restriction.

Value of dipyridamole thallium-201 imaging in noninvasive differentiation of idiopathic dilated cardiomyopathy from coronary artery disease with left ventricular dysfunction.

The purpose of this study was to noninvasively differentiate in patients with reduced global left ventricular function between those with idiopathic dilated cardiomyopathy (IDC) and coronary artery disease (CAD). Clinical features and findings of dipyridamole thallium-201 imaging in 55 consecutive patients with IDC were compared with those in 77 with CAD. Left ventricular ejection fraction was similar between the 2 groups (34 +/- 16% vs 39 +/- 7%). Patients with IDC had lower incidences of ischemic chest pain (11 vs 79%; p less than 0.0001), electrocardiographic evidence of myocardial infarction (24 vs 82%; p less than 0.0001), and reversible defects (4 vs 57%; p less than 0.0001) than did those with CAD. The lowest percent thallium uptake in the initial imaging was less with CAD than IDC (30 +/- 15% vs 59 +/- 10%; p less than 0.001). Patterns of perfusion defects were classified as: no defects, multiple small defects and large defects. Of patients with IDC, 15 had no defects, 19 had multiple small defects, and 21 had large defects, whereas all those with CAD had large defects (p less than 0.0001). Stepwise discriminant analysis, using chest pain and electrocardiography, revealed sensitivity of 89%, specificity of 87%, accuracy of 88%, and positive predictive value of 83% in the identification of patients with IDC.(ABSTRACT TRUNCATED AT 250 WORDS)

Noninvasive identification of significant narrowing of the left main coronary artery by dipyridamole thallium scintigraphy.

To evaluate the usefulness of dipyridamole thallium scintigraphy with low-level exercise for the identification of left main (LM) coronary artery disease (CAD), 466 consecutive patients with CAD were studied. Thirty-eight patients (8%) had LM stenosis (diameter narrowing greater than or equal to 50%). The LM scintigraphic pattern was present in 9 of 38 patients with LMCAD and 38 of 428 CAD patients without LMCAD (24 vs 9%; p less than 0.005). This pattern was present in 6 of 9 patients with LMCAD without right CAD and in only 3 of 29 patients with LM and right CAD (67 vs 10%; p = 0.0005). Patients with LMCAD had a higher incidence of premature cessation of low-level exercise (53 vs 21%; p less than 0.0001), chest pain (68 vs 48%; p less than 0.02), blood pressure decrease of greater than or equal to 20 mm Hg (44 vs 16%; p less than 0.002) and greater ST depression (0.17 +/- 0.13 vs 0.06 +/- 0.10 mV; p less than 0.001) during dipyridamole loading than patients without LMCAD. Stepwise discriminant analysis revealed that the LM scintigraphic pattern and markers of ischemia during dipyridamole loading best identified (p less than 0.0001) patients with LMCAD without right CAD (sensitivity 67%, specificity 91%), but this predictability is no better than the LM scintigraphic pattern alone. The combination of clinical markers of ischemia during dipyridamole loading and scintigraphic findings of diffuse slow washout, extensive fixed defects and the LM pattern best identified (p less than 0.0001) patients with LM and right CAD (sensitivity 72%, specificity 80%).(ABSTRACT TRUNCATED AT 250 WORDS)

Prognostic value of thallium-201 perfusion defects in idiopathic dilated cardiomyopathy.

To assess the prognostic significance of thallium-201 perfusion defects in patients with idiopathic dilated cardiomyopathy (IDC), 43 patients underwent thallium scintigraphy in addition to clinical, echocardiographic, angiographic and hemodynamic evaluation. Eleven patients had no significant thallium perfusion abnormality, 19 had multiple small defects and 13 had a large defect. During 3.2 +/- 2.2 years, 14 patients had disease-related mortality. The patients who died had a higher incidence of ventricular tachycardia (71 vs 31%; p less than 0.02), increased cardiothoracic ratio (60 +/- 6 vs 54 +/- 6; p = 0.005), decreased fractional shortening (11 +/- 6 vs 15 +/- 5; p less than 0.05), increased pulmonary wedge pressure (15 +/- 7 vs 10 +/- 6 mm Hg; p = 0.05), increased left ventricular end-diastolic pressure (21 +/- 8 vs 14 +/- 6 mm Hg; p = 0.02) and abnormal thallium perfusion defects (13 of 14 vs 16 of 26; p less than 0.05) compared with survivors. Age, gender, left ventricular end-systolic and end-diastolic dimensions, cardiac index and ejection fraction were not statistically different in the survivors versus the patients who died. Kaplan-Meier survival estimates at 1, 3 and 5 years were 100% in patients without significant perfusion abnormality; 89, 77 and 64%, respectively, in patients with multiple small defects; and 84, 76 and 30%, respectively, in patients with a large defect (p less than 0.025 by log rank test).(ABSTRACT TRUNCATED AT 250 WORDS)

Age-related changes in amino acid pool sizes in the adult silkmoth, Bombyx mori, reared at low and high temperature; a biochemical examination of the rate-of-living theory and urea accumulation when reared at high temperature.

To examine the rate-of-living theory, age-related changes in amino acid pool sizes were investigated in the adult silkmoth, Bombyx mori, reared at low and high temperature. At either temperature concentrations of free amino acids contained in silkmoths revealed a great sexual difference. Those in females were generally much higher than in males and the former changed much more dynamically than the latter. Major amino acids or ninhydrin-positive compounds inclusive of some essential amino acids such as Leu, Ile, Val, Thr, Arg, Phe, Met, Ala, Tyr, Gln, Aspn , Lan , Cysta , GABA and PEA accumulated in 4 degrees C-moths. However, the levels of these amino changed irregularly with advanced age. Inhibition of protein synthesis may occur generally at low temperature, while protein degradation may be promoted at high temperature. High concentrations of MSO and Tau in the moths reared at high temperature than in the normal moths suggested also catabolism of amino acids proceeding together with protein degradation at high temperature. Amino acid metabolism seems to be complicated under various temperature conditions. When reared at the optimal temperature of 25 degrees C, urea is not present in the body of the silkmoth except for a slight amount in the secreted meconium. In silkmoths reared at the higher temperature of 35 degrees C, however, an extraordinary accumulation of urea occurs accompanied by a reduction in lifespan by one half. Undoubtedly, urea is produced in this terrestrial insect, although the accumulation mechanism is not clear: in silkmoths reared at various temperatures, arginase is found, but urease is not detected. Arginase activity was found to be higher in male moths than in female moths regardless of the rearing temperature. High temperature rearing also did not induce activity and female activity never exceeded that in males at either 25 degrees C or 35 degrees C rearing. Protein degradation accelerated by rearing at high temperatures may result in increased amounts of free arginine, which could cause the active production of urea. This possibility would be a counter-argument to the rate of living theory relating to longevity and temperature. However, at least the above facts signify that an extrinsic factor influences the longevity of an animal by altering its intrinsic aging process.

Congenital coronary arteriovenous fistula associated with dilated cardiomyopathy.

A 59-year-old housewife with heart failure was found to have dilated cardiomyopathy associated with bilateral coronary artery fistula to the pulmonary artery. Coexistence of coronary arteriovenous fistula and dilated cardiomyopathy has not been reported and seems to be a casual association.

Cyclopentadithiophene (CPDT) dimer dication: bipolaron model for quaterthiophenes.

[structure: see text] Cyclopentadithiophene (CPDT) dimers in which both 3,3' and 4' ',3' " positions were bridged with 1,3-dioxalane, carbonyl, or dicyanovinylidene were prepared. These compounds have small HOMO-LUMO gaps (1.03-2.25 eV). The electrochemical oxidation of a dicyanovinylidene-bridged CPDT dimer gave a dication that had a quinoid-like structure.

Antagonism of ceruletide, a cholecystokinin analog, to the neurochemical effects of the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonists, phencyclidine and MK-801, on regional dopaminergic neurons in the rat brain.

In the present study, we investigated the effects of ceruletide (CL), a cholecystokinin analog, on the neurochemical response to non-competitive N-methyl-D-aspartate (NMDA) receptor antagonists, phencyclidine (PCP) and MK-801, of the dopaminergic neuron systems in the discrete regions of the rat brain. Systemically administered PCP (7.5 mg/kg, i.p.) or MK-801 (1.0 mg/kg, i.p.) produced significant increases in the tissue contents of dopamine metabolite, homovanillic acid (HVA), in the prefrontal cortex, the nucleus accumbens and the olfactory tubercle but not in the nucleus caudatus putamen after 60 min. The effects of NMDA receptor antagonists in the nucleus accumbens and the prefrontal cortex were partially antagonized by pretreatment with CL (80 and 400 micrograms/kg, i.p., at 60 min prior to the drugs). While CL alone decreased the dopaminergic metabolism only in the nigrostriatal pathways in naive rats, the present results indicated that CL also attenuates the activities of the meso-limbic and meso-cortical dopaminergic neuron systems when these are enhanced by either PCP or MK-801.

Effect of phencyclidine on dopamine release in the rat prefrontal cortex; an in vivo microdialysis study.

The effect of phencyclidine (PCP) on the extracellular dopamine levels in the rat prefrontal cortex was investigated using an in vivo brain dialysis technique. PCP increased extracellular dopamine levels in the prefrontal cortex of freely-moving rats after the systemic (7.5 mg/kg i.p.) or the local injection (100 microM and 500 microM). The local injection of MK-801, which is a more selective and potent NMDA receptor antagonist than PCP also increased the extracellular dopamine levels (from 10 microM to 100 microM). These results suggest that part of the effect of PCP is attributable to its antagonist effect on the NMDA receptor.