Quantification of HPV16 E7 Oncoproteins in Urine Specimens from Women with Cervical Intraepithelial Neoplasia.

We present the validity of using an ultrasensitive enzyme-linked immunosorbent assay (ELISA) for quantifying high-risk human papillomavirus (HPV) 16 E7 oncoproteins in urine specimens as a noninvasive method of analyzing the oncogenic activity of HPV. Some reports claim that the oncogenic activity of HPV is a more relevant clinical indicator than the presence of HPV DNA for estimating malignant potential. In the present study, urine containing HPV16 and related types were selected by uniplex E6/E7 polymerase chain reaction and classified according to the pathologic diagnosis of cervical intraepithelial neoplasia (CIN) in cervical biopsy specimens. Our ultrasensitive ELISA was able to detect attomole levels of HPV16 E7 oncoproteins, and it detected HPV16-positive SiHa cells at >500 cells/mL without detecting HPV18-positive cells. Our ELISA results showed E7 oncoproteins in 80% (4/5) of urine specimens from women with HPV16-positive CIN1, 71% (5/7) of urine specimens from CIN2 patients, and 38% (3/8) of urine specimens from CIN3 patients. Some urine specimens with undetectable E7 oncoproteins were thought to be negative for live HPV 16-positive cells or in an inactivated state of infection. These results provide the basis for assessing oncogenic activity by quantifying E7 oncoproteins in patient urine.

Dietary supplement use and its related factors among Chinese international and Korean college students in South Korea.

BACKGROUND/OBJECTIVES: The consumption of dietary supplements has shown an increase among young people in their 20s. We aimed to compare the use of dietary supplements and related factors between Chinese international and Korean college students living in South Korea. SUBJECTS/METHODS: We conducted online surveys of 400 Chinese international students and 452 Korean college students from January to February 2021. We analyzed the factors related to the use of dietary supplements by these students using multi-group structural equation modeling and logistic regression analysis. RESULTS: Approximately 65% of the Chinese international students and 93% of the Korean college students consumed dietary supplements at least once in the year preceding the survey. The common types of dietary supplements consumed by both groups of students were vitamin and mineral supplements, Lactobacillus products, and red ginseng products. Structural equation modeling showed that perception of the consumption of dietary supplements by family and friends positively influenced attitude toward dietary supplements. This effect was higher for Korean college students than for Chinese international students (P < 0.01). Attitude toward dietary supplements positively influenced their use, and this effect was higher for Chinese international students than for Korean college students (P < 0.001). Logistic regression analysis showed that the use of dietary supplements by Chinese international students was significantly associated with age, self-reported health status, interest in health, perception of and attitude toward dietary supplements, and length of residence in South Korea. Among Korean college students, it was associated with exercise frequency and attitude toward dietary supplements. CONCLUSION: This study showed significant differences in the use of dietary supplements and related factors between Chinese international and Korean college students. Therefore, nutrition education programs on dietary supplements need to have differentiated content for each group. Such differences also suggest that the industry should consider the relevant characteristics of college students while developing and marketing dietary supplements.

Development of nutrient-based nutritional standards for foodservice at shelters during disasters in the Republic of Korea.

BACKGROUND/OBJECTIVES: In times of disaster, simplified and minimized nutritional standards are necessary for a quick response to provide nutritious relief food. This study aimed to develop nutrient-based nutritional standards for foodservice at shelters during disasters in the Republic of Korea (South Korea). SUBJECTS/METHODS: The standards were developed in 2 phases. First, nutrients to be included in the standards were selected. Initial candidates were selected considering 3 aspects: preceding standards, insufficient intake during disasters, and inadequate intake among South Koreans. Final selection was made by excluding nutrients for 3 reasons: nutrients for which there is no deficiency concern in South Korea, nutrients whose intake data were not available, or nutrients whose values presented by Dietary Reference Intakes for Koreans are difficult to achieve based on the current diet among South Koreans. Second, the reference values of energy and the selected nutrients were calculated. The reference values for the entire population who were 1-year-old and over were calculated by multiplying the estimated energy requirements or the recommended nutrient intake and the proportion of each age and sex group. Respective reference values were also calculated for 4 different age groups (1-5, 6-11, 12-64, and ≥ 65-year-old). RESULTS: The standards for the entire population were 2,000 kcal for energy, 55 g for protein, 650 µg retinol activity equivalents for vitamin A, 95 mg for vitamin C, 1.1 mg for thiamin, 1.3 mg for riboflavin, 14 mg niacin equivalents for niacin, 350 µg dietary folate equivalents for folic acid, 750 mg for calcium, and 11 mg for iron. Four additional standards corresponding to each age group were developed. CONCLUSIONS: The nutritional standards during disasters were developed for South Korea, including energy and 9 nutrients with reference values for the entire population and 4 different age groups. The standards will contribute to maintaining the health of disaster evacuees in South Korea.

Effects of beta-hydroxy beta-methyl butyrate calcium combined with exercise therapy in patients with cardiac disease: a study protocol for clinical trial.

INTRODUCTION: The current treatment for heart disease consists of exercise therapy in addition to pharmacotherapy, nutritional support and lifestyle guidance. In general, nutritional support focuses on protein, salt and energy restrictions, with no active protein or amino acid intake in cases involving moderate or higher renal failure. From this perspective, patients with cardiac disease are at high risk of frailty.Beta-hydroxy beta-methyl butyrate (HMB) is a metabolite of leucine. HMB is widely used for muscle strengthening and can be safely ingested even by patients with renal failure. The proposed study protocol will investigate the effects of HMB-calcium (HMB-Ca) administered in combination with comprehensive cardiac rehabilitation for muscle strength, muscle mass and cardiac function in patients with cardiac disease during the convalescent period. The primary outcome will be knee extensor strength. Secondary outcomes will be gross isometric limb strength and skeletal muscle mass. METHODS AND ANALYSIS: This study will be a single-blinded, randomised, controlled trial with parallel comparisons between two groups. The study period will be 60 days from the start of outpatient cardiac rehabilitation. Participants will be randomly divided into two groups: an HMB group consuming HMB-Ca one time per day for 60 days; and a Placebo group consuming reduced maltose once one time per day for 60 days. Exercise therapy will be performed by both groups. ETHICS AND DISSEMINATION: The study protocol will be published in a peer-reviewed journal. Ethics approval was provided by the Showa University Clinical Research Review Board. TRIAL REGISTRATION NUMBER: jRCTs031220139; Japan Registry of Clinical Trails.

Successful cochlear implantation in a patient with Epstein syndrome during long-term follow-up.

Epstein syndrome is a rare disease characterized by macrothrombocytopenia, nephritis and progressive sensorineural hearing loss (SNHL). This syndrome is presently recognized as an autosomal dominant disease caused by mutations of non-muscle myosin heavy chain 9 (MYH9). Little information is available about the progress of SNHL, the efficacy of cochlear implants (CI) or the perioperative management of thrombocytopenia in patients with Epstein syndrome. We herein report a case of a patient with Epstein syndrome with the MYH9:c.2105G>A:p.R702H variant who underwent cochlear implantation after 27 years of follow-up for her progressive SNHL. The deterioration rates of hearing were 3.48 dB/year on the right ear and 2.46 dB/year on the left ear. The patient derived benefits from CI and had a speech recognition test result (for sentences) of 93% at 6-months postoperatively. Thrombocytopenia was successfully managed without any bleeding complications by using eltrombopag, an oral thrombopoietic agent, making transfusion of platelets unnecessary. The accurate diagnosis of Epstein syndrome was made only after long-term follow-up as the thrombocytopenia was initially diagnosed as idiopathic thrombocytopenic purpura. This case report highlights the perioperative management of thrombocytopenia, the progress of SNHL and the potential pitfalls of diagnosis.

Novel ACTG1 mutations in patients identified by massively parallel DNA sequencing cause progressive hearing loss.

Human ACTG1 mutations are associated with high-frequency hearing loss, and patients with mutations in this gene are good candidates for electric acoustic stimulation. To better understand the genetic etiology of hearing loss cases, massively parallel DNA sequencing was performed on 7,048 unrelated Japanese hearing loss probands. Among 1,336 autosomal dominant hearing loss patients, we identified 15 probands (1.1%) with 13 potentially pathogenic ACTG1 variants. Six variants were novel and seven were previously reported. We collected and analyzed the detailed clinical features of these patients. The average progression rate of hearing deterioration in pure-tone average for four frequencies was 1.7 dB/year from 0 to 50 years age, and all individuals over 60 years of age had severe hearing loss. To better understand the underlying disease-causing mechanism, intracellular localization of wild-type and mutant gamma-actins were examined using the NIH/3T3 fibroblast cell line. ACTG1 mutants p.I34M p.M82I, p.K118M and p.I165V formed small aggregates while p.R37H, p.G48R, p.E241K and p.H275Y mutant gamma-actins were distributed in a similar manner to the WT. From these results, we believe that some part of the pathogenesis of ACTG1 mutations may be driven by the inability of defective gamma-actin to be polymerized into F-actin.

Clinical Characteristics and In Vitro Analysis of MYO6 Variants Causing Late-Onset Progressive Hearing Loss.

MYO6 is known as a genetic cause of autosomal dominant and autosomal recessive inherited hearing loss. In this study, to clarify the frequency and clinical characteristics of hearing loss caused by MYO6 gene mutations, a large-scale genetic analysis of Japanese patients with hearing loss was performed. By means of massively parallel DNA sequencing (MPS) using next-generation sequencing for 8074 Japanese families, we found 27 MYO6 variants in 33 families, 22 of which are novel. In total, 2.40% of autosomal dominant sensorineural hearing loss (ADSNHL) in families in this study (32 out of 1336) was found to be caused by MYO6 mutations. The present study clarified that most cases showed juvenile-onset progressive hearing loss and their hearing deteriorated markedly after 40 years of age. The estimated hearing deterioration was found to be 0.57 dB per year; when restricted to change after 40 years of age, the deterioration speed was accelerated to 1.07 dB per year. To obtain supportive evidence for pathogenicity, variants identified in the patients were introduced to MYO6 cDNA by site-directed mutagenesis and overexpressed in epithelial cells. They were then assessed for their effects on espin1-induced microvilli formation. Cells with wildtype myosin 6 and espin1 co-expressed created long microvilli, while co-expression with mutant constructs resulted in severely shortened microvilli. In conclusion, the present data clearly showed that MYO6 is one of the genes to keep in mind with regard to ADSNHL, and the molecular characteristics of the identified gene variants suggest that a possible pathology seems to result from malformed stereocilia of the cochlear hair cells.

Prevalence and clinical features of hearing loss caused by EYA4 variants.

Variants in the EYA4 gene are known to lead to autosomal dominant non-syndromic hereditary hearing loss, DFNA10. To date, 30 variants have been shown to be responsible for hearing loss in a diverse set of nationalities. To better understand the clinical characteristics and prevalence of DFNA10, we performed genetic screening for EYA4 mutations in a large cohort of Japanese hearing loss patients. We selected 1,336 autosomal dominant hearing loss patients among 7,408 unrelated Japanese hearing loss probands and performed targeted genome enrichment and massively parallel sequencing of 68 target genes for all patients. Clinical information of cases with mutations in EYA4 was gathered and analyzed from medical charts. Eleven novel EYA4 variants (three frameshift variants, three missense variants, two nonsense variants, one splicing variant, and two single-copy number losses) and two previously reported variants were found in 12 probands (0.90%) among the 1,336 autosomal dominant hearing loss families. The audiometric configuration of truncating variants tends to deteriorate for all frequencies, whereas that of non-truncating variants tends to show high-frequency hearing loss, suggesting a new correlation between genotype and phenotype in DFNA10. The rate of hearing loss progression caused by EYA4 variants was considered to be 0.63 dB/year, as found in this study and previous reports.

Relationship Between Performance Improvement in Activities of Daily Living and Energy Intake in Older Patients With Hip Fracture Undergoing Rehabilitation.

OBJECTIVE: To analyze whether sufficient energy intake (EI) improves performance of activities of daily living (ADL) in patients with hip fracture admitted to rehabilitation hospitals. The adequate amount of EI for improving performance of ADL in patients with hip fracture remains unknown. METHODS: This retrospective cohort study included all patients with hip fracture (n=234) admitted to rehabilitation hospitals in Japan. The inclusion criteria for this study were age >65 years and body mass index <30.0 kg/m2. Patients who were transferred to an acute hospital and those with missing case data were excluded. According to the amount of EI, the patients were classified into energy sufficiency and shortage groups (EI/total energy expenditure ≥1.0 and <1.0, respectively). The Functional Independence Measure (FIM) and FIM gain were used to evaluate the patient disability level and change in patient status in response to rehabilitation. Finally, FIM gain was calculated as the discharge FIM score minus the admission FIM score. RESULTS: The final analysis targeted 202 patients-53 (26.2%) were in the energy shortage group and 149 (73.8%) were in the energy sufficiency group. The energy sufficiency group had a greater FIM gain than the energy shortage group (mean, 25.1±14.2 vs. 19.7±16.4; p=0.024). Furthermore, sufficient EI in the first week since admission (ß=0.165; 95% confidence interval, 0.392-5.230; p=0.023) was an independent factor of FIM gain. CONCLUSION: Among elderly patients with hip fracture admitted to rehabilitation hospitals in Japan, the amount of EI during the first week after admission was an independent factor of FIM gain.

Mutational Spectrum and Clinical Features of Patients with LOXHD1 Variants Identified in an 8074 Hearing Loss Patient Cohort.

Variants of the LOXHD1 gene, which are expressed in hair cells of the cochlea and vestibule, have been reported to cause a progressive form of autosomal recessive non-syndromic hereditary hearing loss, DFNB77. In this study, genetic screening was conducted on 8074 Japanese hearing loss patients utilizing massively parallel DNA sequencing to identify individuals with LOXHD1 variants and to assess their phenotypes. A total of 28 affected individuals and 21 LOXHD1 variants were identified, among which 13 were novel variants. A recurrent variant c.4212 + 1G > A, only reported in Japanese patients, was detected in 18 individuals. Haplotype analysis implied that this variation occurred in a mutational hot spot, and that multiple ancestors of Japanese population had this variation. Patients with LOXHD1 variations mostly showed early onset hearing loss and presented different progression rates. We speculated that the varying severities and progression rates of hearing loss are the result of environmental and/or other genetic factors. No accompanying symptoms, including vestibular dysfunction, with hearing loss were detected in this study. Few studies have reported the clinical features of LOXHD1-gene associated hearing loss, and this study is by far the largest study focused on the evaluation of this gene.

The Prevalence and Clinical Characteristics of TECTA-Associated Autosomal Dominant Hearing Loss.

TECTA is well known as a causative gene for autosomal dominant mid-frequency hearing loss observed in various populations. In this study, we performed next-generation sequencing analysis of a large Japanese hearing loss cohort, including eight hundred and twelve (812) subjects from unrelated autosomal dominant hearing loss families, to estimate the prevalence and phenotype-genotype correlations in patients with TECTA mutations. The prevalence of TECTA mutations in Japanese autosomal dominant sensorineural hearing loss families was found to be 3.2%. With regard to the type of hearing loss, the patients with mutations in the nidogen-like domain or ZA domain of TECTA showed varied audiograms. However, most of the patients with mutations in the ZP domain showed mid-frequency hearing loss. The rate of hearing deterioration in TECTA-associated hearing loss patients and in the normal hearing Japanese control population were the same and regression lines for each group were parallel. We carried out haplotype analysis for four families which had one recurring missense variant, c.5597C>T (p.Thr1866Met). Our results revealed four different haplotypes, suggesting that this mutation occurred independently in each family. In conclusion, TECTA variants represent the second largest cause of autosomal dominant sensorineural hearing loss in Japan. The hearing loss progression observed in the patients with TECTA mutations might reflect presbycusis. The c.5597C>T mutation occurred in a mutational hot spot and is observed in many ethnic populations.

Mid-Frequency Hearing Loss Is Characteristic Clinical Feature of OTOA-Associated Hearing Loss.

The OTOA gene (Locus: DFNB22) is reported to be one of the causative genes for non-syndromic autosomal recessive hearing loss. The copy number variations (CNVs) identified in this gene are also known to cause hearing loss, but have not been identified in Japanese patients with hearing loss. Furthermore, the clinical features of OTOA-associated hearing loss have not yet been clarified. In this study, we performed CNV analyses of a large Japanese hearing loss cohort, and identified CNVs in 234 of 2262 (10.3%, 234/2262) patients with autosomal recessive hearing loss. Among the identified CNVs, OTOA gene-related CNVs were the second most frequent (0.6%, 14/2262). Among the 14 cases, 2 individuals carried OTOA homozygous deletions, 4 carried heterozygous deletions with single nucleotide variants (SNVs) in another allele. Additionally, 1 individual with homozygous SNVs in the OTOA gene was also identified. Finally, we identified 7 probands with OTOA-associated hearing loss, so that its prevalence in Japanese patients with autosomal recessive hearing loss was calculated to be 0.3% (7/2262). As novel clinical features identified in this study, the audiometric configurations of patients with OTOA-associated hearing loss were found to be mid-frequency. This is the first study focused on the detailed clinical features of hearing loss caused by this gene mutation and/or gene deletion.

OTOF mutation analysis with massively parallel DNA sequencing in 2,265 Japanese sensorineural hearing loss patients.

The OTOF gene (Locus: DFNB9), encoding otoferlin, is reported to be one of the major causes of non-syndromic recessive sensorineural hearing loss, and is also reported to be the most common cause of non-syndromic recessive auditory neuropathy spectrum disorder (ANSD). In the present study, we performed OTOF mutation analysis using massively parallel DNA sequencing (MPS). The purpose of this study was to reveal the frequency and precise genetic and clinical background of OTOF-related hearing loss in a large hearing loss population. A total of 2,265 Japanese sensorineural hearing loss (SNHL) patients compatible with autosomal recessive inheritance (including sporadic cases) from 53 otorhinolaryngology departments nationwide participated in this study. The mutation analysis of 68 genes, including the OTOF gene, reported to cause non-syndromic hearing loss was performed using MPS. Thirty-nine out of the 2,265 patients (1.72%) carried homozygous or compound heterozygous mutations in the OTOF gene. It is assumed that the frequency of hearing loss associated with OTOF mutations is about 1.72% of autosomal recessive or sporadic SNHL cases. Hearing level information was available for 32 of 39 patients with biallelic OTOF mutations; 24 of them (75.0%) showed profound hearing loss, 7 (21.9%) showed severe hearing loss and 1 (3.1%) showed mild hearing loss. The hearing level of patients with biallelic OTOF mutations in this study was mostly severe to profound, which is consistent with the results of past reports. Eleven of the 39 patients with biallelic OTOF mutations had been diagnosed with ANSD. The genetic diagnosis of OTOF mutations has significant benefits in terms of clinical decision-making. Patients with OTOF mutations would be good candidates for cochlear implantation; therefore, the detection of OTOF mutations is quite beneficial for patients, especially for those with ANSD.

Frequency and clinical features of hearing loss caused by STRC deletions.

Sensorineural hearing loss is a common deficit and mainly occurs due to genetic factors. Recently, copy number variants (CNVs) in the STRC gene have also been recognized as a major cause of genetic hearing loss. We investigated the frequency of STRC deletions in the Japanese population and the characteristics of associated hearing loss. For CNV analysis, we employed a specialized method of Ion AmpliSeqTM sequencing, and confirmed the CNV results via custom array comparative genomic hybridization. We identified 17 probands with STRC homozygous deletions. The prevalence of STRC homozygous deletions was 1.7% in the hearing loss population overall, and 4.3% among mild-to-moderate hearing loss patients. A 2.63% carrier deletion rate was identified in both the hearing loss and the control population with normal hearing. In conclusion, our results show that STRC deletions are the second most common cause of mild-to-moderate hearing loss after the GJB2 gene, which accounts for the majority of genetic hearing loss. The phenotype of hearing loss is congenital and appears to be moderate, and is most likely to be stable without deterioration even after the age of 50. The present study highlights the importance of the STRC gene as a major cause of mild-to-moderate hearing loss.

Diagnostic pitfalls for GJB2-related hearing loss: A novel deletion detected by Array-CGH analysis in a Japanese patient with congenital profound hearing loss.

Here, we report a novel deletion (copy number variation: CNV) in the GJB2 gene observed in a Japanese hearing loss patient. The deleted segment started in the middle of the GJB2 gene, but the GJB6 gene remained intact. This partial deletion in the GJB2 gene highlights the need for further improvements in GJB2 screening.

Sensorineural hearing loss and mild cardiac phenotype caused by an EYA4 mutation.

EYA4 is a member of the vertebrate eya gene family of transcriptional activators and plays several roles in both embryonic and inner ear development. The majority of EYA4 gene mutations are associated with autosomal dominant non-syndromic hearing loss (DFNA10). In addition, some mutations in this gene cause autosomal dominant syndromic hearing loss with dilated cardiomyopathy. EYA4 is a rare cause of sensorineural hearing loss, and only a limited number of papers regarding mutations in this gene have been published. Thus, detailed clinical features remain unclear. We conducted next-generation sequencing of a Japanese individual with progressive sensorineural hearing loss and identified an EYA4 pathogenic variant. Pure-tone audiometry revealed bilateral, nearly symmetric, moderate sensorineural hearing loss in the low and middle frequencies. Minor abnormalities were observed on the patient's electrocardiogram and echocardiography without any apparent symptoms. Next-generation sequencing is effective in elucidating the etiology of hearing loss, and the present findings suggested the possible phenotypic expansion of deafness caused by EYA4 gene mutations.

Nutritional quality and patterns of lunch menus at child care centers in South Korea and Japan.

BACKGROUND AND OBJECTIVES: This study aimed to investigate the nutritional quality and patterns of lunch menus provided by child care centers in South Korea and Japan. METHODS AND STUDY DESIGN: The weekly lunch menus from Monday to Saturday that child care centers provided in November 2014 in South Korea and Japan were analyzed. For Korea, a total of 72 meals provided by 12 centers in Seoul were analyzed by referring to the homepage of the Center for Children's Foodservice Management, which serviced menus for child care centers. For Japan, a total of 30 meals provided by 5 child care centers in Tokyo were analyzed. Nutrient content and pattern in lunch menus were evaluated. RESULTS: The lunch menus in Korea and Japan provided 359.5 kcal (25.7% of the estimated energy requirement) and 376.3 kcal (29.5% of the estimated energy requirement), respectively. 'Rice + Soup + Main dish + Side dish I + Side dish II' were provided in 66.7% of meals in Korea, while various patterns with rice and soup as their bases were provided in Japan. CONCLUSIONS: The lunch menus of child care centers in Korea and Japan provide similar amounts of energy, protein, carbohydrate, vitamin A, calcium, and other nutrients. However, there were significant differences in the lunch menu patterns in Korea and Japan. This study provides information about the nutritional content and pattern of lunch menus at child care centers in Asian countries with rice as a staple food.

Clinical evaluations of complete autologous fibrin glue, produced by the CryoSeal® FS system, and polyglycolic acid sheets as wound coverings after oral surgery.

The CryoSeal® FS System has been recently introduced as an automated device for the production of complete fibrin glue from autologous plasma, rather than from pool allogenic or cattle blood, to prevent viral infection and allergic reaction. We evaluated the effectiveness of complete autologous fibrin glue and polyglycolic acid (PGA) sheet wound coverings in mucosa defect oral surgery. Postoperative pain, scar contracture, ingestion, tongue dyskinesia, and postoperative bleeding were evaluated in 12 patients who underwent oral (including the tongue) mucosa excision, and received a PGA sheet and an autologous fibrin glue covering. They were compared with 12 patients who received a PGA sheet and commercial allogenic fibrin glue. All cases in the complete autologous fibrin glue group demonstrated good wound healing without complications such as local infection or incomplete cure. All evaluated clinical measures in this group were similar or superior to the commercial allogenic fibrin glue group. Coagulation and adhesion quality achieved with this method was comparable to that with a PGA sheet and commercial fibrin glue. Covering oral surgery wounds with complete autologous fibrin glue produced by an automated device was convenient, safe, and reduced the risk of viral infection and allergic reaction associated with conventional techniques.

Comparison of the nutrient-based standards for school lunches among South Korea, Japan, and Taiwan.

Nutritional standards are important guidelines for providing students with nutritionally-balanced school meals. This study compared nutrient-based school lunch standards regulated by South Korea, Japan, and Taiwan. The data were collected from relevant literature and websites of each country during September 2014. The number of classification groups of target students was 8, 5, and 5 for South Korea, Japan, and Taiwan, respectively. Gender was considered across all age groups in South Korea but only for high school students in Taiwan. Gender was not considered in Japan. Along with energy, the number of nutrients included in the standards for South Korea, Japan and Taiwan was 9, 12, and 4, respectively. The standards for all three countries included protein and fat among macronutrients. The standards for South Korea and Japan included vitamin A, B-1, B-2, and C, while the standards for Taiwan did not include any vitamins. Calcium was the only mineral commonly included in the three standards. The proportions of recommended daily intakes as reference values for each nutrient differed among the countries. Japan differentiated the proportions among 33%, 40%, or 50%, reflecting the target students' intake status of the respective nutrients. Taiwan differentiated either two-fifths or one-third of the recommended daily intakes. South Korea applied the proportion of recommended daily intake as one-third for all selected nutrients. This study could be valuable information for countries in developing nutrient-based standards for school lunches and for South Korea, Japan, and Taiwan in the process of reforming nutrient-based standards.

Detection of human papillomavirus infection in oral squamous cell carcinoma: a cohort study of Japanese patients.

OBJECTIVE: Recent evidence suggests that human papillomavirus (HPV)-related head and neck squamous cell carcinoma (HNSCC) is a separate HNSCC subgroup with distinct epidemiology, histopathological characteristics, therapeutic response to chemotherapy and radiation, and clinical outcome. This study aimed to investigate the role of HPV infection in oral squamous cell carcinoma (OSCC) and the correlation between HPV infection, tumor suppressor protein p16 expression, and clinicopathological features in Japanese patients. METHODS: In total, 174 OSCC specimens were examined for p16 levels by immunohistochemistry, and p16-positive OSCCs were analyzed for HPV DNA by in situ hybridization (ISH) and HPV genotypes by real-time PCR. The results were evaluated for the association with clinicopathological characteristics of OSCC patients. RESULTS: Twenty-four OSCC samples were found positive for p16 expression; all of them were well-differentiated tumors. P16 immunoreactivity was significantly associated with the invasion depth and tended to correlate with sex, site in the oral cavity, stromal reaction, TNM stage, and survival. HPV DNA was detected in 13 of 24 (54%) p16-positive OSCC by real-time PCR; HPV 16, 18, and other high-risk genotypes were the most prevalent. However, ISH failed to detect HPV DNA in p16-positive OSCCs. CONCLUSION: P16 immunoreactivity and HPV genotyping by real-time PCR may be useful markers of HPV infection in OSCC. However, although HPV-related OSCC showed good outcomes, HPV infection may have a minor role in oral oncogenesis in Japanese patients.