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Biomedical subjects

Yuko Morikawa

Publications and source records attributed to Yuko Morikawa.

At least 37 records · Page 2Linked to original sources

Binding site for fungal beta-lactone hymeglusin on cytosolic 3-hydroxy-3-methylglutaryl coenzyme A synthase.

We studied the molecular mechanism through which the fungal beta-lactone, hymeglusin, potently and specifically inhibits 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase. [(14)C]Hymeglusin covalently bound to purified rat liver and to recombinant hamster cytosolic HMG-CoA synthases. The enzyme activity was completely inhibited at a binding ratio of 1.6-2.0 mol [(14)C]hymeglusin/mol HMG-CoA synthase. Incubating the enzyme with 2 mM iodoacetamide (IAA) or 2 mM N-ethylmaleimide (NEM) but not with 1.0 mM diisopropyl fluorophosphates (DFP) completely inhibited the binding, suggesting that hymeglusin binds to a Cys residue of HMG-CoA synthase. Recombinant hamster HMG-CoA synthase labeled with [(3)H]hymeglusin was digested with V8 protease, and the [(3)H]peptide was purified by high performance liquid chromatography (HPLC). The sequence of the peptide was Ser-Gly-Asn-Thr-Asp-Ile-Glu-Gly-Ile-Asp-Thr-Thr-Asn-Ala-[(3)H]hymeglusyl Cys-Tyr-Gly-Gly-Thr-Ala-Ala-Val-Phe-Asn-Ala-Val-Asn-, which corresponds to the active site sequence (from Ser 115 to Asn 141) of hamster HMG-CoA synthase. These findings showed that hymeglusin inhibits hamster cytosolic HMG-CoA synthase by covalently modifying the active Cys 129 residue of the enzyme.

Amino Acid Sequence↗

Mortality in a cadmium polluted area in Japan.

A 15-year follow-up study of 3178 inhabitants (1424 men and 1754 women) living in the cadmium (Cd) polluted Kakehashi River basin was conducted. The results clarified effects on mortality of renal dysfunction induced by Cd indicated by urinary beta-2-microglobulin (beta2-MG), total protein, glucose, and total amino acids. This study used Cox's proportional hazard model. The mortality risk ratio of urinary beta2-MG positive (>= 1000 microg/gCr) subjects was significantly increased in both sexes: 1.35 for men and 1.73 for women. The increased mortality ratio of the urinary protein positive (>= 10 mg/dl) subjects was also significant for both sexes, with risk ratios of 1.82 for men and 2.01 for women. Only the women showed significantly increased mortality of the urinary glucose positive (>= 20 mg/dl) subjects and amino acids positive (> = 300 microg/gCr) subjects. When the subjects were divided into four categories according to urinary beta2-MG, <300, 300-1000, 1000-10000, >= 10000 microg/gCr, the mortality risk ratios were increased in proportion to the increase of urinary beta2-MG in both sexes. These results suggest that mortality of Cd-exposed subjects increased with increasing excretion of four urinary markers of renal tubular dysfunction, and in proportion to increases in the amount of urinary beta2-MG excretion including under 1000 microg/gCr.

Amino Acids↗

Lack of association between human G-protein beta3 subunit variant and overweight in Japanese workers.

We examined the association of the G-protein beta3 subunit gene (GNB3) C825T polymorphism with overweight in Japanese workers. This cross-sectional study used multivariate analysis to investigate whether a polymorphism in the C825T polymorphism was associated independently with overweight when factors such as age and lifestyle were taken into account. The study in 1453 men and 1172 women involved identifying subjects with the C825T genotype using the polymerase chain reaction, followed by restriction fragment-length polymorphism analysis. Overweight was defined as a BMI > or = kg/m(2). Genotype distributions for C825T in overweight men (CC = 80, CT = 162, TT = 80) and women (CC = 52, CT = 91, TT = 40) were not significantly different from normal-weight men (CC = 278, CT = 588, TT = 265) and women (CC = 242, CT = 549, TT = 198). The allele distributions were also not significantly different between either sex. The power of the study was estimated as 98% in men and 81% in women based on the allelic frequencies reported in a previous positive study in Chinese subjects. Multiple logistic regression analysis showed that the genotype was not significantly associated with overweight. In conclusion, this study indicated that the GNB3 C825T polymorphism is not a significant factor for overweight in Japanese people.

Adult↗

A comparison of socio-economic differences in long-term sickness absence in a Japanese cohort and a British cohort of employed men.

OBJECTIVES: To compare the magnitude of socio-economic differences in sickness absence rates between a Japanese cohort and a British cohort. To assess the effects of self-rated health and behavioural risk factors on sickness absence in the two cohorts, and whether they explain socio-economic differences in sickness absence within and between cohorts. METHODS: An 8 year follow up study of sickness absence in 2504 Japanese male employees in a factory in Japan and 6290 British male employees in civil service departments in London. The rates of first occurrences of long-term (>7 calendar days) sickness absence were determined and compared between these cohorts. Socio-economic status was measured with hierarchical employment grades. RESULTS: The first time sickness absence rates were about two times higher among British men as compared with Japanese men. The rate ratio of lower to higher employment grade was 1.2, 1.3 and 2.1 among Japanese white-collar, Japanese blue-collar and British white-collar employees respectively. Baseline self-rated health and smoking habit predicted sickness absence in both cohorts. After adjusting for these factors a significant difference between the Japanese and British cohorts, and between employment grades remained. CONCLUSIONS: The rate of long-term sickness absence was higher in the British cohort than the Japanese cohort.

Absenteeism↗

Lack of association of human G-protein beta 3 subunit variant with hypertension in Japanese workers.

OBJECTIVE: To examine the association between hypertension and the C825T polymorphism in the G-protein beta 3 subunit gene in Japanese workers. METHODS: This study used logistic regression analysis and multiple regression analyses to investigate whether the C825T polymorphism was associated independently with hypertension or blood pressure when factors such as age, body mass index, blood chemistry and lifestyle were taken into consideration. The target subjects were 1452 male and 1169 female workers selected from 3834 male and 2591 female workers in a single company. Hypertension was defined as a systolic blood pressure > or = 140 mmHg and/or diastolic blood pressure > or = 90 mmHg, or taking antihypertensive medication. The power of the study was estimated as 83% for males and 41% for females based on allelic frequencies in Caucasians. RESULTS: Genotype distributions for C825T in hypertensive males (CC = 58, CT = 135, TT = 63) and females (CC = 20, CT = 36, TT = 20) were not significantly different from normotensive males (CC = 300, CT = 614, TT = 282) or females (CC = 274, CT = 602, TT = 217), respectively. Allele distributions were not significantly different in either sex. Multiple logistic regression analysis showed that genotype was not associated significantly with hypertension, whereas there was a significant relationship between hypertension and age, family history of hypertension, body mass index, hematocrit, platelet count, gamma-glutamyl transpeptidase (gamma-GTP) and uric acid. Data analysis using one-way analysis of variance and multiple regression showed that the C825T allele had no significant influence on either systolic, diastolic or mean blood pressure. CONCLUSIONS: This study indicates that the C825T polymorphism is not a significant factor for hypertension or blood pressures in Japanese people. Targeting of this polymorphism is therefore unlikely to be beneficial when attempting to prevent hypertension in the general Japanese population.

Adult↗

Environmental factors and risk of idiopathic dilated cardiomyopathy: a multi-hospital case-control study in Japan.

BACKGROUND: Detailed epidemiological investigations on the relationship of environmental factors, especially occupational and microbiological factors, to the development of idiopathic dilated cardiomyopathy (IDC) are scarce. METHODS AND RESULTS: A multi-hospital case-control study was conducted in 38 hospitals throughout Japan in order to survey IDC cases and age, sex-matched outpatient controls at each hospital. Crude and adjusted odds ratios (ORs) by various environmental factors were calculated in 135 pairs of cases and controls. Univariate analyses revealed significantly increased ORs for lower education, passive smoking in the workplace, cold and/or hot workplace, symptoms of fatigue and history of bacterial infection; in contrast, decreased ORs were associated with a history of rubella and gastroduodenal diseases. Based on multivariate adjusted analyses, lower education (OR 1.96, 95% confidence interval (CI) 1.13-3.40), cold or hot workplace (OR 1.84, 95%CI 1.08-3.12) and history of measles (OR 1.78, 95%CI 1.01-3.08) exhibited a significant positive relationship with IDC risk. History of rubella (OR 0.17, 95%CI 0.06-0.52) and gastroduodenal diseases (OR 0.14, 95%CI 0.07-0.29) were inversely related to the risk. CONCLUSIONS: Some occupational and microbiological factors appear to relate independently to the development of IDC and further investigation is required to establish their respective mechanisms.

Adult↗

The influence of work characteristics on body mass index and waist to hip ratio in Japanese employees.

A cross-sectional study on 6,676 workers consisting of 4,243 males and 2,433 females aged 20-58 yr in a metal product factory was conducted to elucidate the relationship between work characteristics, e.g. job demand/control/support, sedentary job, overtime work and shift work, and waist to hip ratio (WHR) as well as body mass index (BMI) taking alcohol consumption, smoking, exercise and other psychosocial factors such as education and marital status into account. By a stepwise multiple regression analysis, BMI was associated with shift work, marital status and sedentary job for males, and with exercise but inversely associated with education for females. WHR was also associated with shift work, alcohol consumption, marital status and sedentary job but inversely associated with exercise for males, and with sedentary job, marital status and education but inversely associated with smoking for females. These results suggest that work characteristics such as sedentary job and shift work should also be considered when trying to prevent increases in BMI and WHR.

Adult↗

High-density lipoprotein cholesterol and risk of stroke in Japanese men and women: the Oyabe Study.

BACKGROUND AND PURPOSE: Evidence of an inverse relationship between serum high-density lipoprotein cholesterol (HDL-C) and the risk of stroke is sparse in Asians and in women. The purpose of this investigation was to examine the relationship in a long-term cohort study of Japanese men and women among whom stroke occurrence is higher than in Western countries. METHODS: A prospective cohort study was performed involving 4989 participants (1523 men, 3466 women) 35 to 79 years of age at baseline with approximately 10 years of follow-up in a rural area of Japan. End points included all stroke incidence and ischemic stroke incidence. RESULTS: During follow-up, 132 participants developed stroke, including 81 ischemic stroke cases. Age-adjusted incidence rates per 10,000 person-years for all stroke in subjects with low HDL-C (<30 mg/dL [0.78 mmol/L]) were 103.4 in men and 49.3 in women, which were remarkably higher than in subjects with high HDL-C (>or=60 mg/dL [1.56 mmol/L]) (26.4 in men and 15.5 in women). A similar relationship was observed for ischemic stroke. Multivariate-adjusted relative risks for all stroke incidence and ischemic stroke incidence were 2.89 (95% CI, 1.35 to 6.20) and 2.92 (95% CI, 1.17 to 7.32), respectively, for low versus high HDL-C participants. The relationships were independent of sex, age, body mass index, blood pressure, serum total cholesterol, alcohol consumption, and smoking. CONCLUSIONS: This 10-year follow-up study of Japanese men and women demonstrated that lower HDL-C levels were related significantly and independently to increased risk of all stroke incidence and ischemic stroke incidence.

Adult↗

Time course of Gag protein assembly in HIV-1-infected cells: a study by immunoelectron microscopy.

Recent biochemical studies have identified high molecular complexes of the HIV Gag precursor in the cytosol of infected cells. Using immunoelectron microscopy we studied the time course of the synthesis and assembly of a HIV Gag precursor protein (pr55gag) in Sf9 cells infected with recombinant baculovirus expressing the HIV gag gene. We also immunolabeled for pr55gag human T4 cells acutely or chronically infected with HIV-1. In Sf9 cells, the time course study showed that the first Gag protein appeared in the cytoplasm at 28-30 h p.i. and that budding started 6-8 h later. Colloidal gold particles, used to visualize the Gag protein, were first scattered randomly throughout the cytoplasm, but soon clusters representing 100 to 1000 copies of pr55gag were also observed. By contrast, in cells with budding or released virus-like particles the cytoplasm was virtually free of gold particles while the released virus-like particles were heavily labeled. Statistical analysis showed that between 80 and 90% of the gold particles in the cytoplasm were seen as singles, as doublets, or in small groups of up to five particles probably representing small oligomers. Clusters of gold particles were also observed in acutely infected lymphocytes as well as in multinuclear cells of chronically infected cultures of T4 cells. In a few cases small aggregates of gold particles were found in the nuclei of T4 lymphocytes. These observations suggest that the Gag polyprotein forms small oligomers in the cytoplasm of expressing cells but that assembly into multimeric complexes takes place predominantly at the plasma membrane. Large accumulations of Gag protein in the cytoplasm may represent misfolded molecules destined for degradation.

Animals↗

Trend of congenital anomalies over 20 years ascertained by population-based monitoring in Ishikawa Prefecture, Japan.

Monitoring in Ishikawa Prefecture (mean population; 1,150,000), during the 20-year period from 1981 to 2000, involved monitoring the prevalence of congenital anomalies starting at gestational week 22. The main objectives of the present study were (1) to investigate the trend over the past 20 years in the prevalence of congenital anomalies, and (2) to investigate whether there is area accumulation in congenital anomalies or not. In addition, the prevalence of well-known risk factors such as smoking, drinking and family history among cases with congenital anomalies was analyzed to obtain clues to the background of such anomalies. During the 20 years, more than 200,000 births have been observed. Thirty-three selected congenital anomalies, consisting mainly of external anomalies, were analyzed in this study. The percentage of children born with congenital anomalies did not change markedly between the first and second halves of the monitoring period, remaining at approximately 4.5/1,000 births. There was also no significant change in annual trends in either Ishikawa Prefecture as a whole or in the three regions within Ishikawa. The prevalence of congenital anomalies by type showed a significant decrease in anencephaly and spina bifida. In contrast, Down syndrome and microcephaly increased significantly. The mean maternal age for Down syndrome increased significantly from the first-half period to the second-half period. The percentage of children with polydactyly whose mother smoked were relatively higher than that for the other congenital anomalies.

Congenital Abnormalities↗

Yeast-derived human immunodeficiency virus type 1 p55(gag) virus-like particles activate dendritic cells (DCs) and induce perforin expression in Gag-specific CD8(+) T cells by cross-presentation of DCs.

To evaluate the immunogenicity of human immunodeficiency virus (HIV) type 1 p55(gag) virus-like particles (VLPs) released by budding from yeast spheroplasts, we have analyzed the effects of yeast VLPs on monocyte-derived dendritic cells (DCs). Yeast VLPs were efficiently incorporated into DCs via both macropinocytosis and endocytosis mediated by mannose-recognizing receptors, but not the mannose receptor. The uptake of yeast VLPs induced DC maturation and enhanced cytokine production, notably, interleukin-12 p70. We showed that yeast membrane components may contribute to DC maturation partly through Toll-like receptor 2 signaling. Thus, Gag particles encapsulated by yeast membrane may have an advantage in stimulating Gag-specific immune responses. We found that yeast VLPs, but not the control yeast membrane fraction, were able to activate both CD4(+) and CD8(+) T cells of HIV-infected individuals. We tested the effect of cross-presentation of VLP by DCs in two subjects recruited into a long-term nonprogressor-slow progressor cohort. When yeast VLP-loaded DCs of these patients were cocultured with peripheral blood mononuclear cells for 7 days, approximately one-third of the Gag-specific CD8(+) T cells were activated and became perforin positive. However, some of the Gag-specific CD8(+) T cells appeared to be lost during in vitro culture, especially in a patient with a high virus load. Our results suggest that DCs loaded with yeast VLPs can activate Gag-specific memory CD8(+) T cells to become effector cells in chronically HIV-infected individuals, but there still remain unresponsive Gag-specific T-cell populations in these patients.

Antigen Presentation↗

HIV capsid assembly.

HIV Gag assembly is the first and most essential step in the formation of virus particles. Following protein synthesis, Gag relocates from ribosomes and forms a virus particle at the plasma membrane, using host factors and machinery. Early studies focused on mapping the regions within Gag required for assembly and identified three distinct domains (M, I, and L), although their precise locations within the three-dimensional structure of Gag awaited later study. In this review, I summarize the mapping results in the light of recent progress on Gag structures made by nuclear magnetic resonance and X-ray crystallography as well as further functional analysis. These data are largely consistent and provide sufficient information for an understanding of the interactions and functions of the assembly domains at a macromolecular level. Current studies have moved on to the identification of the host factors and machinery used in the process of Gag assembly. Cumulative data suggest that the dynamics of Gag assembly and transport are achieved not by simply using, but rather by taking control of, cellular machinery. Key area in the process include interactions with TSG101, L domain receptor which normally functions in the endosomal sorting pathway and with lipid rafts, a type of M domain receptor, which has been suggested to be the sites for effective concentration of Gag. The review provides a summary of these data and discusses the likely direction of future studies.

Capsid Proteins↗

[Relationship of childhood obesity to adult obesity: a 20-year longitudinal study from birth in Ishikawa Prefecture, Japan].

OBJECTIVE: Evidence regarding the relationship between childhood obesity and adult obesity in Japan is limited. This study was conducted to determine the relationship between childhood mass index (BMI) at 3 months, 12 months, 3 years and 20 years in a general population. METHODS: Data obtained from men and women aged 20 years (born between 1968-1974), who had received medical examinations in Ishikawa Prefecture, Japan, were linked to data of medical examinations of the same individuals as infants (3 months, 12 months, and 3 years). The relationship between childhood BMI (or Kaup index) and adult BMI was analyzed for a total of 2,314 participants (1,080 men and 1,234 women), whose data could be followed for 20 years. RESULTS: BMI at 20 years displayed significant positive correlations with BMI at 3 months, 12 months, and 3 years; this correlation was strongest with respect to BMI at 3 years (r = 0.33, p < 0.001 in men; r = 0.42, p < 0.001 in women). In terms of percentages of obese participants (BMI 25 kg/m2 or over) at 20 years in accordance with BMI categories at each age, the rates were 4.6% in men and 1.0% in women with a BMI less than 15 kg/m2 at 3 years, but 29.1% and 29.5%, respectively, with a BMI of 18 kg/m2 or over (6.3 and 29.5 times higher, respectively). Percentages of obese participants at 20 years were highest in those exhibiting an above average BMI at 3 years, regardless of the BMI at 3 months. CONCLUSIONS: Body mass in young adults is strongly related to body mass in childhood, especially with that at 3 years. About 30 percent of obese children at 3 years remain obese into adulthood. These results are of interest with respect to assessment of future risk of adulthood obesity at medical examinations for infants in Japan.

Adult↗

Identification of Hsp90 as a stimulatory host factor involved in influenza virus RNA synthesis.

Efficient transcription and replication of the influenza virus genome are dependent upon host-derived factors. Using an in vitro RNA synthesis system, we have purified and identified Hsp90 as one of the host factors that stimulate viral RNA polymerase activity. Hsp90 interacted with the PB2 subunit of the viral RNA polymerase through the amino-terminal chaperone domain and the middle region containing a highly acidic domain. The acidic middle region was also responsible for its stimulatory activity. We found that a portion of Hsp90 is re-localized to the cell nucleus after viral infection. A PB2 fragment containing a Hsp90 binding domain inhibited viral gene expression in a dominant-negative manner. These results suggest that Hsp90 is a host factor for the influenza virus RNA polymerase.

Amino Acid Sequence↗

HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomyces cerevisiae.

Expression of retroviral Gag protein in yeast has previously shown Gag targeting to the plasma membrane but little or no production of Gag virus-like particles (VLPs). Here we show that, after removal of the cell wall, the expression of HIV type 1 Gag protein in Saccharomyces cerevisiae spheroplasts allowed simultaneous budding of VLPs from the plasma membrane. Our data show that (i) the VLPs released from yeast spheroplasts were spherical and had morphological features, such as membrane apposed electron-dense layers, characteristic of the immature form of HIV particles; (ii) the VLPs were completely enclosed in the plasma membrane derived from yeast, which is denser than that of higher eukaryotic cells; (iii) the VLP Gag shells remained intact after treatment of nonionic detergent; and (iv) the VLPs were released soon after removal of the cell wall and accumulated up to 300 microg/liter of culture. Our results also show that VLP production was abolished by amino acid substitution of the Gag N-terminal myristoylglycine and impaired when Gag C-terminal deletions were extended beyond the nucleocapsid domain. These results were consistent with those obtained previously in higher eukaryotic expression systems, suggesting that similar Gag domains were used for VLP assembly. We suggest that the system described here offers significant advantages for studying host factors required for VLP budding. The system also may be available for production of vector virus-free VLPs for practical applications such as vaccine development.

Base Sequence↗

Epidemiologic and clinical characteristics of cardiomyopathies in Japan: results from nationwide surveys.

Nationwide clinico-epidemiological surveys of cardiomyopathies in Japan were carried out. Disorders surveyed included idiopathic dilated cardiomyopathy (DCM), hypertrophic cardiomyopathy (HCM), restrictive cardiomyopathy (RCM), arrhythmogenic right ventricular dysplasia (ARVD), mitochondrial disease, Fabry's disease of the heart and prolonged Q-T interval syndrome. The total number of patients was estimated at 17,700 for DCM, 21,900 for HCM, 300 for RCM, 520 for ARVD, 640 for mitochondrial disease, 150 for Fabry's disease of the heart, and 1,000 for prolonged Q-T interval syndrome. The prevalence of both DCM and HCM was higher in men than women: the male-to-female ratios were 2.6 and 2.3 for DCM and HCM, respectively. Detailed data on patients with DCM or HCM were collected by a follow-up survey. In 1 year more patients with DCM (5.6%) died than with HCM (2.8%): congestive heart failure (CHF) and arrhythmias were the leading causes of death for DCM and HCM, respectively. Angiotensin converting enzyme inhibitors (64.6%) and beta-adrenergic blockers (40.9%) are commonly used to treat the CHF complicating DCM and may be associated with the clinical improvement in a significant number of DCM patients. Thus, the nationwide surveys of Japanese patients have yielded important current epidemiological and clinical information on the characteristics of cardiomyopathies in Japan.

Adolescent↗

Effects of fatty liver induced by niacin-free diet with orotic acid on the metabolism of tryptophan to niacin in rats.

The effects of dietary orotic acid on the metabolism of tryptophan to niacin in weaning rats was investigated. The rats were fed with a niacin-free, 20% casein diet containing 0% (control diet) or 1% orotic acid diet (test diet) for 29 d. Retardation of growth, development of fatty liver, and enlargement of liver were observed in the test group in comparison with the control group. The concentrations of NAD and NADP in liver significantly decreased, while these in blood did not decrease compared to the control group. The formation of the upper metabolites of tryptophan to niacin such as anthranilic acid, kynurenic acid, and 3-hydroxyanthranilic acid were not affected, but the quinolinic acid and beyond, such as nicotinamide, N1-methylnicotinamide, N1-methyl-2-pyridone-5-carboxamide, and N1-methyl-4-pyridone-3-carboxamide, were significantly reduced by the administration of orotic acid. Therefore, the conversion ratio of tryptophan to niacin significantly decreased in the test group in comparison with the control group.

Animals↗

[The effects of nutrition and life-style on calcaneal bone mass in high school students].

OBJECTIVE: To clarify the effects of nutrient/food intake and lifestyle, especially exercise, on calcaneal bone mass in both male and female high school students. METHODS: A total of 798 healthy Japanese high school students (281 boys and 517 girls) were recruited, and their calcaneal bone was assessed by ultrasound using Lunar Achilles. The subjects were thereby divided into a lower bone mass group and a normal bone mass group according to stiffness at the cut-off level of 90 for the boys, and 77 for the girls. Information on lifestyle including frequency of food consumption and regular exercise during the period at junior high school and high school was collected by questionnaire. Nutrient intake was assessed by a semi-quantitative food frequency questionnaire. RESULTS: 1. The intake of total energy and fat in the lower bone mass group was lower than in the normal bone mass group in both sexes. 2. The frequency of subjects not consuming milk in the lower bone mass group was higher than in the normal bone mass group in both sexes. In the boys, the frequency of the subjects who liked to eat green vegetables in the lower bone mass group was lower in the normal bone mass group. 3. The mean stiffness for subjects undertaking exercise during junior high school was higher than in subjects not exercising, even after adjustment for age in both sexes. The frequency of subjects with a habit of sun bathing in the lower bone mass group was significantly lower than in the normal bone mass group in the girls. 4. An analysis using a logistic model was performed to select the factors related to lower bone mass without being confounded with other factors. Lower energy intake in boys and lower fat intake, no exercise during high school life and no milk consumption in girls were each independently related to lower bone mass. CONCLUSION: These results suggest that nutrition and exercise habit affect calcaneal bone mass in high school students of both sexes, especially milk consumption being an important factor related to bone mass in girls.

Adolescent↗