To determine the prevalence and effect of dietary habits on functional constipation in preschool and early elementary school children in Japan.  A total of 3595 children aged 3 to 8 years from 28 nursery schools and 22 elementary schools in Yokohama City, Kanagawa Prefecture, Japan, were evaluated. The subjects were divided into a functional constipation group and a nonfunctional constipation group according to the Rome III criteria. Dietary intake data were collected using a brief-type, self-administered, diet-history questionnaire validated for Japanese preschool-aged children. p = 0.617). A decrease in bowel frequency was observed in 15.9% of those with functional constipation. There was no significant difference in the proportion of participants in the constipation group by age (p = 0.112). Binomial logistic regression analysis indicated that only fat per 100 kcal positively correlated with functional constipation .  Of the 3595 subjects evaluated, 718 (20.0%) had functional constipation. The association between functional constipation and gender was not statistically significant ( Functional constipation is common among children in preschool and early elementary school in urban areas of Japan. Parents should pay attention to constipation-related symptoms other than defecation frequency. A high-fat diet should be avoided to prevent functional constipation. Constipation is one of the most common health problems in children and adults. Variation in the normal bowel habit and symptoms recognized as constipation has made it difficult to accurately diagnose the condition in children. In addition, overflow incontinence and encopresis, both symptoms of functional constipation (FC), are often considered simply as problematic bowel habits and not as symptoms of FC. Therefore, it is difficult to clarify the current state of FC in childhood. The Rome III criteria , 2 have It is well known that dietary habit can affect defecation, and as such, regional disparity should be considered when determining the prevalence of constipation. Although increasing the intake of dietary fibre and water is often recommended as a dietary therapy for patients with constipation, the influence of total calories and individual nutrients on FC in childhood have not been clarified. A brief-type, self-administered, diet-history questionnaire (BDHQ) developed by Kobayashi et al. has been validated for dietary intake assessment in adults . Asakura2.1.Two types of questionnaire were distributed to the guardians of 2052 children attending 28 public or private nursery schools and 4451 children attending 1st or 2nd grade at 22 public elementary schools  in Tsurumi Ward, Yokohama City, Kanagawa Prefecture, Japan, in September 2013. The questionnaires were collected between October and November 2013. One questionnaire assessed defecation and physical status, and the other was the BDHQ3y, which assessed total calories consumed and the intake of individual nutrients. The questionnaires were enclosed and concealed in envelopes by each guardian. The envelopes were collected at each school and sent back to the authors by a representative at each school.This survey included questions on sex, age, height, weight, defecation frequency, fecal incontinence, withholding behaviour, painful defecation, stool consistency, and the presence of large-diameter stools. Guardians were required to answer questions regarding defecation frequency, stool consistency using the Bristol Stool Scale type, and frequency at 4 grades  for other details related to defecation. Guardians had to choose the Bristol Stool Scale type that best represented their children's stools using a picture chart. The picture chart was accompanied by descriptors that had been translated into Japanese.The BDHQ3y is a four-page structured questionnaire that asks about the consumption frequency of selected foods commonly eaten in Japan, general dietary behaviour, and usual cooking methods. Estimates of the daily intake of different foods , total calories, and selected nutrients were calculated using an ad hoc computer algorithm for the BDHQ3y based on the Standard Tables of Food Composition in Japan . AlthougThe defecation status of each subject was classified according to the Rome III criteria . Questio2). Height and weight were evaluated using z-scores (standard deviation (SD) scores). The z-score is the deviation of an individual's value from the median value of a reference population, divided by the standard deviation of the reference population . The z-scores (SD scores) for height and weight were calculated according to the standard formula z = (sample data − mean)/SD. z-Scores for height and weight and the percentile of BMI were calculated using the taikakushisu_v3.xlsx spreadsheet  developed by the Standard Value Committee of The Japanese Society of Pediatric Endocrinology/The Japanese Association for Human Auxology.Height, body weight, and body mass index (BMI) were evaluated based on the report of a national growth survey in preschool children  and an aThe BDHQ3y calculates the intake amounts of 99 different types of food nutrients, including total calories, meal weight, water, protein, fat, carbohydrates, 15 food groups, and 78 ingredients based on an assessment of food-intake frequency per month. Inputting questionnaire data and conversion of the quantities of calories, nutrients, and food groups consumed were performed by the developer's company , and we then used the converted data for analysis. We analysed 15 items, including total amount per day of meal weight, total calories, water content in foods, protein, fat, carbohydrates, calcium, magnesium, phosphorus, soluble dietary fibre, insoluble dietary fibre, total dietary fibre, sodium chloride, juice, and water including tea. Twelve items, excluding juice and water, were analysed as the nutritional equivalent per 100 kcal.This survey was approved by the Ethics Committee of Saiseikai Yokohama City Tobu Hospital (protocol number 201230). Response to the questionnaire was regarded as an agreement to participate, in accordance with Ethical Guidelines for Epidemiological Research published by the Ministry of Health, Labor and Welfare.χ2 test. Associations between FC and physical status, such as height and weight and energy and nutrient intake, were initially examined using t-tests, and analysis of covariance was performed with age as a covariate to compare the FC group and non-FC group. Then, binomial logistic regression analysis was performed to identify factors independently associated with FC in children. All statistical analyses were performed using SPSS statistical software . Two-sided p values < 0.05 were considered statistically significant.The prevalence of FC and the symptoms detailed in the Rome III criteria were examined using Pearson's n = 3595) are shown in p = 0.617) or in age (p = 0.112) between the two groups. However, the average age of the girls in the FC group was significantly higher than that of the girls in the non-FC group . The mean body weight and the mean BMI were higher in the FC group than the non-FC group , but in the analysis using age as a covariate, only the mean BMI percentile value was significantly higher in the FC group than the non-FC group (p = 0.042). Numbers of obesity (BMI percentile > 90) were 59 (8.2%) in the FC group and 186 (6.5%) in the non-FC group, but there was no significant difference between the two groups (p = 0.059). There was no significant difference in the proportion of children in the constipation group by age (p = 0.290). Binomial logistic regression analysis did not identify any factors associated with FC.A total of 3932 guardians returned the questionnaires which gave a collection rate of 58.7%. Overall, 3643 subjects answered both questionnaires. Of these, 14 subjects aged 9 years, 3 subjects who did not give their age, and 31 subjects who gave incomplete descriptions were excluded from further analysis. Thus, in total, 3595 subjects were analysed in this study. The characteristics of the analysed children , with odds ratios as high as 24.5 and 11.0, respectively, and high sensitivity and specificity. The incidences of the items “≤2 defecations per week” and “history of large-diameter stools that obstruct the toilet” in the FC group were low , although the specificity for both was 0.98.p = 0.005) and greater juice intake  but less intake of water, including tea  (p = 0.003), meal water content , magnesium intake , soluble dietary fibre intake , insoluble dietary fibre intake , total dietary fibre intake , and sodium chloride intake  than the non-FC group. In contrast, the intake of fat was significantly higher in the FC group than in the non-FC group . Binomial logistic regression analysis indicated that only fat per 100 kcal positively correlated with FC   . Similar6–1.412] .We could analyse 25.4% of the population of children aged 3 to 8 years in the survey area. Hence, our results reflect the prevalence of FC among children aged 3 to 8 years living in urban areas in Japan, which is approximately 20% regardless of age or sex. It should be noted that because the item in the Rome III criteria that asks about the presence of a large fecal mass in the rectum was not included in the questionnaires in this survey, the prevalence of childhood FC may be higher.Asakura et al. conducted a school-based survey to examine the relationship between constipation and lifestyle factors, including dietary intake, among preschool-aged children in Japan . The parIn this study, a history of excessive stool retention and painful bowel movements were observed in 73.6% and 61.5% of the children in the FC group. In contrast, only 15.9% of the children in the FC group experienced 2 or less defecations per week. In previous studies, such differences between items in the Rome III criteria were not observed , 20. In p = 0.042), whereas there was no significant difference in the number of obese children between the two groups (p = 0.059). In normal individuals after a high-fat meal, abnormal gastrocolic reflex with prolonged retrograde phasic contraction was observed . In mouse experiments, feeding of a high-fat diet resulted in intestinal dysbiosis and delayed colonic motility . Howeverobserved . In rodeobserved , 27. Theobserved . MoreoveThe main limitations of this study were that the Japanese questionnaire on defecation was not validated before its use and the question about the presence of large fecal mass in the rectum was omitted in the questionnaire. These factors could have influenced the prevalence of FC obtained in this study.The current findings suggest that FC is common among children in preschool and early elementary school in urban areas in Japan. Decreased frequency of bowel movements is not a common symptom of FC in this population, and guardians should pay attention to other symptoms, including withholding defecation and painful defecation. Consumption of a high-fat diet should be avoided to prevent FC. A longitudinal study and an expanded study of additional age groups are also needed to determine whether FC in children aged 3 to 8 years persists into late childhood, adolescence, and adulthood.


Drosophila sperm experiments using high dose radiation delivered in a short period. The fallacious 60 years-old LNT is applied to cancer induction by radiation without solid data and then to cancer induction by carcinogens also without solid data. Therefore, even the smallest amount of carcinogens is postulated to be carcinogenic without thresholds now. Radiation hormesis is observed in a large variety of living organisms; radiation is beneficial at low doses, but hazardous at high doses. There is a threshold at the boundary between benefit and hazard. Hormesis denies LNT. Not a few papers report existence of chemical hormesis. If mutagens and carcinogens show hormesis, the linear dose-response relationship in mutagenesis and carcinogenesis is denied and thresholds can be introduced.The Japanese Environmental Mutagen Society (JEMS) was established in 1972 by 147 members, 11 of whom are still on the active list as of May 1, 2016. As one of them, I introduce some historic topics here. These include 1) establishment of JEMS, 2) the issue of 2-(2-furyl)-3-(3-nitro-2-furyl)acrylamide (AF-2), 3) the Mammalian Mutagenicity Study Group (MMS) and its achievements, and 4) the Collaborative Study Group of the Micronucleus Test (CSGMT) and its achievements. In addition to these historic matters, some of which are still ongoing, a new collaborative study is proposed on adaptive response or hormesis by mutagens. There is a close relationship between mutagens and carcinogens, the dose-response relationship of which has been thought to follow the linear no-threshold model (LNT). LNT was fabricated on the basis of  When the members of the board of trustees of the Japanese Environmental Mutagen Society (JEMS) were asked if they would contribute their papers to Environmental Mutagen Research (EMR), the former title of Genes and Environment (G&E), when they write papers, more than half answered no. They would rather contribute them to Mutation Research, Mutagenesis, or Environmental and Molecular Mutagenesis. The main reason was that papers in EMR were not cited in PubMed. When I was the editor-in-chief of EMR (1998–1999), I tried to make EMR a PubMed-citation journal, but my trials and efforts came to naught. EMR was a quarterly magazine and consisted of Japanese and English articles. A total annual number of original papers might not reach the criteria of citation by PubMed. EMR was changed to Genes and Environment (G&E), a fully English journal, in 2006. Minako Nagao, editor-in-chief, made a great contribution to G&E. In spite of her ardent efforts, however, citation in PubMed was not successful. The editor-in-chief was replaced by Takashi Yagi in 2011, and M. Nagao became the production editor. As she retired from this position, I took over the job from her in 2014. As the production editor, I looked through all of the accepted papers and checked them for errors and for format, so as not to deviate from the instructions to authors. One comment from me is the deletion of keywords from the title so as to avoid duplication. G&E became an open access journal in 2015, and I was freed from the position of production editor. Recently, I was delighted to learn that PubMed decided to cite G&E papers. From my viewpoint, this is a dream come true and more congratulatory than the 10th anniversary itself. As I am one of JEMS members from the start, let me make some remarks on the occasion of the PubMed citation and the 10th anniversary of G&E.JEMS was established at the National Education Center in Tokyo on August 21, 1972. The number of participants at the first meeting was 147. Active members are 11 as of May 1, 2016. They are Hikoya Hayatsu, Yasumoto Kikuchi, Taijiro Matsushima, Tohru Shibuya, Hiroyasu Shimada, Takashi Sugimura, Noriho Tanaka, Hiroshi Tanooka, Hideo Tezuka, Makoto Umeda, and I. At first, JEMS was started as the Environmental Mutagen Research Association for the first 6 years. The program and the list of participants at the first meeting were in my hands; I offered them to the JEMS’ office so as not to be lost.Neurospora crassa, Sohei Kondo (Osaka University): Molecular mechanisms of mutations, Takeo Suzuki : Hazardous substances in human environments, Tsuneo Kada : Screening methods for chemical mutagens based on the theories of chemical mutagenesis, Yasuhiko Shirasu : Toxicity issues of pesticides, Hideya Endo (Kyushu University): Chemical carcinogenesis and mutation, Mamoru Saito (Tokyo University): Roles of natural carcinogens in the environment, Fuminori Yanagisawa : Consideration of teratogenicity by alkylbenzene sulfonate, Tsuyoshi Kajiwara : Mutagenesis testing and teratogenicity, Ujihiro Murakami : relationship between teratogens, mutagens, and carcinogens, and W.W. Nichols and R.C. Miller : Anaphase as a cytogenetic method in mutagenicity testing.The organizer-in-chief of the first JEMS meeting was Yataro Tajima, who gave an opening address at 9:30. It was followed by a congratulatory address by E.B. Freese, the then president of the American Environmental Mutagen Society. Fourteen papers were presented at the meeting. Each speaker presented his paper for 25 min. It may be of interest to learn what the top researchers at that time spoke of. Japanese titles are translated into English. Hikoyuki Yamaguchi (Tokyo University): Chromosomal aberrations by antibiotics in plants, Hidetoshi Yoshida and Yukimasa Shiraishi : Chromosomal aberrations by cadmium in humans, Shigeo Iwahara : Mutation induction in bacteria by food-associated substances, F.J. de Serres : Mutation induction in radiation-sensitive strains of A close relationship between mutagenicity and carcinogenicity was a major concern in these days. Carcinogenicity testing was time-consuming, labor-intensive, and costly. Researchers made efforts to develop detection methods for mutagens as substitutes for detection of carcinogens. As a result, a widely used food additive was found to be a mutagen and this created a social problem as shown below.E. coli B/rWP2 Trp−, the inability to detect AF-2 mutagenicity with S. typhimurium strains TA1535, TA1536, TA1537 and TA1538 [Mutagenicity and carcinogenicity was the major issue of AF-2 in the 2nd JEMS meeting, organized by Yataro Tajima and held at the National Institute of Genetics in Mishima City in 1973. Seven papers out of 17 were associated with AF-2. Some insisted that AF-2 was not a carcinogen and others refuted this assertion. There was a very hot debate. AF-2 is an amorphous reddish powder. AF-2 is a stable wide-spectrum antibiotic, and bacteria do not attain tolerance or resistance to it. It is effective at very low dose levels and long-lasting. In 1965, AF-2 was substituted for nitrofurazone, a food additive approved in 1950, and used widely in foods such as hams, sausages, tofu, sweetened bean paste, and so forth. AF-2 would be an ideal food preservative if it lacked mutagenic and carcinogenic properties. The carcinogenic potential of AF-2 was tested, and negative results were obtained in Donryu rats and ICR/JCL mice , and AF-50 is 475 mg/kg in mice at 7-day observation. AF-2 at a concentration of 0.2% in the diet (1/3 of LD50) for up to 2 years was not carcinogenic [50) for 18 months [Critical remarks were that AF-2 induced forestomach cancer, but humans do not have the forestomach as rodents do, and that doses of AF-2 were so high that albino mice were colored by AF-2. The oral LDIt is not too much to say that JEMS’ activities are largely supported by those of subgroups such as the Mammalian Mutagenicity Study Group (MMS), the Bacterial Mutagenicity Study Group, and the Study Group of Mutation Mechanisms. The Environmental Epigenomics Society and the Study Group of Non-mutagenic Carcinogens have suspended their activities at the present time. Since I was involved in the establishment of MMS, let me have a say.MMS was established in 1982 by merging the Dominant Lethal Test Seminar  and the Micronucleus Test Research Association . An ad hoc committee held a meeting in February 1982, and adopted the name of MMS. The objectives of MMS were to scrutinize in vivo mutagenesis testing systems and, by supporting development of the research area, to contribute to safety evaluation in humans. At the meeting held at the occasion of the 11th annual meeting of JEMS in October 1982, in Shuzenji, organized by Yukiaki Kuroda, action programs were adopted and sub-committees were formed: 1) to do collaborative studies, 2) to have workshops, and 3) to examine protocols. MMS decided to have biannual meetings. MMS had the 69th meeting in December 2016, while JEMS had the 45th meeting in this November 2016.As for 1) to do collaborative studies, a spot test group and a micronucleus test (MN) group were organized at first. Collaborative studies by MMS have expanded to more than 20 studies as follows: (1) Mouse spot test, (2) Sex-related difference in MN, (3) Strain difference in MN, (3) Administration route difference in MN, (4) Treatment times in MN, (5) Peripheral blood MN using supravital staining with acridine orange, (6) MN using IARC  carcinogen, (7) Aging and spontaneous and induced MN, (8) Rat peripheral blood MN, (9) Examination of genotoxicity using transgenic animals, (10) Development of MN using organs other than the bone marrow such as the liver, intestine, skin, and gonads, (11) Risk assessment of genotoxicity with special reference to mitomycin C, (12) Development of MN coupled with the 4-week repeat dosing test, (13) Toxicogenomics using mice and rats, (14) relationship between in vivo and in vitro genotoxicity and carcinogenicity (ongoing), (15) In vitro MN using human cells, (16) Development of MN using the liver of juvenile animals, (17) Examination of in vitro and in vivo comet assays, (18) Conduct of an international collaborative in vivo comet assay using rats, (19) Development of MN using the liver and digestive tracts of mature rats given repeated doses (ongoing), and (20) Development of Pig-assay (ongoing). The results of these collaborative studies have been reported in more than 100 papers.As for 2) to have workshops, we had a workshop on how to judge and detect spots using the positive control of stuffed mice before conducting the mouse spot test. Kiyoshi Tutikawa was the main instructor. Several workshops were carried out before the conducts of MN. For example, bone marrow samples from several mice treated with mitomycin C were combined and stained by Makoto Hayashi and slides were distributed to 25 participants. Results are shown in Fig. 1 of my review paper . AlthougA workshop on chromosomal aberrations was also held. On the basis of this workshop, an atlas book was published . ApproxiMMS had its own journal, MMS Communications (MMSC) (ISSN 0918–5976). This was started as a kind of newsletter. MMSC No. 1 was prepared on the occasion of the 14th MMS meeting in November 1988. It was then published annually until 1991 (No. 4). In 1992, Nos. 5 and 6 were issued for the biannual MMS meetings. The first original paper was published by Naohiko Higashikuni and Shizuyo Sutou (ITOHAM FOODS Inc.) in No. 7 issued in 1993 . The NosMy colleagues and I published 10 papers in MMSC . After 1As mentioned above, a small group to examine MN protocols was established at the Shuzenji meeting in 1982. Members were Hiroyasu Shimada (chief), Makoto Hayashi, Yoshisuke Nishi (Japan Tobacco Inc.), Tohru Shibuya, Noriho Tanaka, and myself. We examined 9 guidelines including those of European Economic Community (EEC), US Environmental Protection Agency (EPA), Federal Republic of Germany (FRG), International Commission for Protection Against Environmental Mutagen and Carcinogen (ICPEMC), Japanese Ministry of Health and Welfare (JMHW), Organisation for Economic Co-operation and Development (OECD), and United Kingdom Environmental Mutagenesis Society (UKEMS). Close examination of these protocols for about 2 years revealed that these were not based on experimental data, but were deduced mainly from chromosomal aberration tests. As a result, we decided to carry out collaborative studies and to make MN protocols that are based on data.I proposed to conduct a collaborative study to examine sex-related differences at first, because animal numbers could be halved if there were no sex-differences or if the differences were negligible. This proposal was suggested by my research into sex-determination mechanisms conducted at Susumu Ohno’s laboratory in City of Hope Research Institute in 1980–1981. The sex difference  was follThe 5th ICEM was held at Case Western University, Cleveland, Ohio, on July 10–15, 1989. I had a chance to present our large body of MN data from CSGMT. The audience seemed to be deeply impressed by our large body of data. I think this provided a precedent for data-based protocols and for international experimental collaborations thereafter. After my presentation, Motoi Ishidate, Jr. told me that I won fame internationally. I answered, not me but CSGMT.China had planned to hold the 6th ICEM in 1993; however, the crackdown known as the Tiananmen Square Massacre occurred on June 4, 1989. Chinese troops with rifles and tanks killed unarmed Chinese civilians in Beijing. The ICEM board decided not to have the 6th ICEM in China, saying that an international meeting could not be held in such a barbarous county. So the 6th ICEM was held in Melbourne in 1993, hosted by the Australia and New Zealand Environmental Mutagen Society (ANZEMS). The Chinese Environmental Mutagen Society hosted the International Symposium on Environmental Mutagenesis and Carcinogenesis in Shanghai in May 1991. I was an invited speaker and presented a paper, “Strain difference in the micronucleus induction among different strains of mice with a special reference to MS/Ae mice.”F.J. de Serres, an editor of Mutation Research, asked me via Motoi Ishidate, Jr. to review the achievements of CSGMT. Earlier topics described above are shown more precisely in this review paper published in 1996 . This miMMS is one of sub-groups of JEMS and consists of around 150 members. CSGMT is one of sub-groups of MMS that carries out MN. The number of members varies depending on subjects. MMS has biannual meetings and results obtained by CSGMT and other groups are reported there.Workshops held before collaborative studies were useful to propagate standard techniques, which supported the acquisition of qualified data.A body of qualified data contributed to the establishment of domestic and international protocols and guidelines, e.g., for the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) and OECD. Subjects associated with regulatory sciences were of great concern for industries, constituting a factor to invite participants from industries.A large number of participants made it possible to collect lots of data in a limited period; the burden to each participant was not so heavy and one could take part in collaborative studies fairly easily. Collaboration among governmental, academic, and industrial researchers was readily achieved.The results of collaborative studies were published in journals. The publication provided the participants with credit. I am not sure, but around 20 people might get the doctorate degree using results of collaborative studies in part.The domestic activities of MMS and CSGMT were integrated, at least in part, into the international framework of guideline setting.This section is written mainly in the past tense. But activities of MMS and CSGMT are still ongoing, and the situation after changing from the past to the present tense would mostly hold.Drosophila melanogaster [Muller discovered that X-rays can induce mutations in nogaster . Atomic nogaster . He defeStandard Oil Co. Inc. was established by John Rockefeller in 1870. The Rockefeller Foundation (RF) was threatened by the discovery of atomic energy. In 1954, RF chose to finance six projects to evaluate atomic radiation. RF asked the U.S. National Academy of Sciences (NAS) to organize the whole program, which was conducted under the auspices of Bronk, president of the Rockefeller University, president of NAS, and an RF trustee. The Genetics Panel (GP) was established in NAS in 1954 and was chaired by Weaver, an RF officer. GP consisted of 17 members, 13 of whom were geneticists including Muller. Most members believed that all doses of radiation were harmful, irreversible, cumulative, and linearly acting, no significant discussion occurred . GP recoDrosophila sex linked recessive lethal test and LNT was fabricated on the data observed in insect sperm that lack repair systems. Actually, responses to X-rays in Drosophila were not linear and showed thresholds and hormesis [X-rays induced mutations at high doses in the hormesis . Lewis chormesis . The NatThe Environmental Protection Agency of the U.S.A. (EPA) is involved in the regulation of carcinogens under several laws. As risk cannot be eliminated completely, EPA introduced a concept of balance between risks and benefits . For risExposure doses are largely underestimated because the doses were estimated only for initial radiation, within 1 min after the atomic bomb blast. The residual radiation was neglected. It was twice as high as the initial radiation and was carried to the ground by black rain. Its effects must have lasted for days or weeks. This means that cancer risk in the LSS is largely overestimated.“In-the-city-control” people, who entered Hiroshima or Nagasaki after the atomic bomb blast and were used as the negative control, were exposed to residual radiation and would not be appropriate as the negative control. Indeed, their cancer mortality rate was less than that of people in the villages northwest of Hiroshima because the “in-the-city-control” people showed hormetic effects .Both reports say that the linear-quadratic fit is better than the linear fit . The Biological Effect of Ionizing Radiations (BEIR) of NAS asserted that the dose-response relation in cancer risk was linear and supported LNT . The latDrosophila sperm, led fallaciously to LNT without relevant data. The failed germ cell mutation hypothesis was applied to somatic cell mutations by radiation without relevant data. LNT is now fundamental basis for radiation-regulatory guidelines. Then, LNT was applied to chemical carcinogenesis without relevant data. Thus, the smallest amount of carcinogens is proportionally carcinogenic on the basis of unproven inference. The linear dose-response relationship of chemical carcinogens without thresholds must be reevaluated.Thus, the LSS does not support LNT. Gene mutations, induced by high-dose radiation in Radiation or chemicals are beneficial at low doses, but hazardous at high doses.Exposure to low doses beforehand gives enhanced cell repair after exposure to high doses.Exposure to X-rays gives tolerance to γ-rays. Exposure to substance A beforehand gives tolerance to substance B afterwards.Recently, I commented on hormesis . AdaptivHormesis must be applied to chemicals. Mutagens and carcinogens would not follow the manner that LNT predicts. Many papers indicate the existence of chemical hormesis. I proposed a new collaborative study on hormesis in mutagenesis at the 68th MMS meeting on June 17, 2016. Presently, a test chemical that is found to be mutagenic in one of the mutagenicity testing methods, it is branded as a mutagen. My expectation is that it might be mutagenic at higher doses, but would be antimutagenic at lower doses. In between higher and lower doses, there must be thresholds. My proposal is to examine the situation experimentally. When mutagenic hormesis is proven, a mutagen will be classified as a hormetic mutagen, indicating that the mutagen is not always hazardous. Many drug candidates might have been dropped off as mutagens during developmental process, but hormetic mutagens could be developed as drugs with less fear about mutation or cancer induction. Changes of guidelines for registration of drugs, cosmetics, pesticides, and so on are expected in the future if hormetic mutagenesis is proven.


A new R software package, RAC, is presented. RAC allows to simulate the rearing cycle of 4 species, finfish and shellfish, highly important in terms of production in the Mediterranean Sea. The package works both at the scale of the individual and of the farmed population. Mathematical models included in RAC were all validated in previous works, and account for growth and metabolism, based on input data characterizing the forcing functions—water temperature, and food quality/quantity. The package provides a demo dataset of forcings for each species, as well as a typical set of husbandry parameters for Mediterranean conditions. The present work illustrates RAC main features, and its current capabilities/limitations. Three test cases are presented as a proof of concept of RAC applicability, and to demonstrate its potential for integrating different open products nowadays provided by remote sensing and operational oceanography. Virtual technologies are increasingly perceived as a resource for aquaculture science-based management . The selDicentrarchus labrax), Gilthead seabream (Sparus aurata), Manila clam (Ruditapes philippinarum) and Mediterranean mussel , both at the individual and the population level. These mathematical models account for growth and metabolism of the individual, based on input data characterizing the forcing functions—water temperature, and food quality and quantity. For finfish, seabass and seabream, food availability is represented by the feed provided by the farmer, while shellfish models require a characterization of quantity and quality of the suspended organic matter. The package provides a demo dataset of forcings for each species, as well as a typical set of husbandry parameters for Mediterranean conditions, in order to allow the user to run a default simulation. The present work illustrates the main features of RAC and delineates its current capabilities in terms of predicting growth and environmental interactions of aquaculture. RAC outputs represent a baseline which could be integrated with environmental spatial data, economic-social criteria and policy issues, in order to support the decision makers in the identification of suitable area for aquaculture activities and for the implementation of an EAA t is time [d]-1]A is the anabolic rate [J d-1]C is the catabolic rate [J d-1] represents the energy density of body tissues.ε , metabolic rates , faeces produced , uneaten feed , O2 consumption  and ammonia production . Mussel model computes the somatic and gonadic dry weight of the individual (g), the wet weight of soft tissues (g), and the total weight including the shell (g), the length (cm), faeces and pseudofaeces produced , metabolic rates (anabolism and catabolism J d-1), CNP content of animal tissues (g), temperature limitation function, O2 consumption  and ammonia production . Clam model outputs include the wet weight of the individual (g), temperature limitation function, metabolic rates (J d-1), and shell length (mm). The number of individuals is also provided as an output. Finally, the number of days needed to reach the commercial size from the beginning of the integration period is reported for all the models.RAC simulations generate two main classes of outputs: i) vector outputs, providing a time series of the number of individuals in the farmed population; ii) matrix outputs, with the mean and the standard deviation of each model state variable, as well as metabolic processes, which are estimated over the distribution of simulated individuals. These output variables, which are different for the 4 models, are summarized in e.g.: computes detritus + zooplankton C concentration as the difference between the POC concentration and phytoplanktonic C concentration). Additionally, the pre-processor function plots the interpolated forcing values, saving them into the previously created folder structure. The pre-processor can be executed by the main script as many times as required in order to satisfy the user requirements. Subsequently, MAIN function calls all the functions that are required to solve the balance and to save the results. The RK-SOLVER function implements the 4th order Runge-Kutta scheme for the solution of the bioenergetics balance. Equations describing limitation terms, and anabolic and metabolic rates are contained in the EQUATIONS function, called by the RK-SOLVER function. Outputs of the RK-SOLVER are processed by the POSTPROCESSOR function that plots and saves them in files. The population script contains also the LOOP function that runs the Monte Carlo simulation and computes the statistics of the simulated outputs. Finally, the POPULATION function solves the population equation, based on mortality rate parameters and on husbandry practices.The general workflow of RAC is schematically described in RAC package requires the subsequent execution of three instructions from the R console. The first instruction creates a directory structure at the path specified by the string variable “userpath”. This instruction populates the directory with a ready to use dataset of forcings and parameters, which can be subsequently modified by the user. The second instruction loads the list of model forcings into the R workspace contained at the user defined path. The third instruction runs the model with the loaded forcings and saves the textual and graphic results into the directories specified by the string “userpath”. In order to select the species to model, and the level of hierarchical organization , two parts of the string need to be specified by the user, while the third part reports the function of interest. In the first part of the string, the user indicates the species , while in the second part indicates “_ind_” or “_pop_”, in order to run the individual or the population model respectively.As an example, in order to run the Gilthead seabream individual model, the following instructions need to be specified:Bream_ind_skeleton (userpath)forcings <–Bream_ind_dataloader (userpath)Output <–Bream_ind_main On the other hand, running the Mussel population model will require the following instructions:Mussel_pop_skeleton (userpath)forcings <–Mussel_pop_dataloader (userpath)Output <–Mussel_pop_main In order to demonstrate the applicability of RAC, and the resources potentially provided by operational oceanography and remote sensing data for running the model, the following set of simulations was performed:individual models were run for the 4 species, using as input present environmental data, taken from the sites marked in mussel individual model was run repeatedly over a spatial domain which represents a portion of sea localized in a highly productive coastal area, in the Northern Adriatic Sea, Emilia-Romagna region—model runs were used to estimate the growth performance of the mussel over this area, which was quantified as days required by the mussel for reaching the length of 5 and 7 cm;-m day-1, were run via Monte Carlo simulation. This number of model runs was empirically found to be the minimum needed to stabilize the results [water temperature time series for 2049–2050, predicted by climate models, were used to run the seabream population model at a site located in the southern Adriatic Sea, Puglia region see . These s results . Farm ch results , Pastres results  and Brig results ,14,15. Ea (Chl-a) data were extracted from the Copernicus—Marine Environment Monitoring Service (http://marine.copernicus.eu/). For the purposes of this study, satellite data level 4 with a spatial resolution of 0.008 (SST) and 0.013 degrees (Chl-a) (~1 Km), from 01/03/2015 to 30/09/2016 were downloaded. As previously described, the shellfish models require TSM, POC and POM concentrations as forcing variables but, since satellite TSM, POC and POM were not available for the study area, these values were imposed on the basis of existing data collected at mussel farms in the area [The SST and Chlorophyll-the area ,18.3 metric tons in 2013, 33.6% of the national production [The area selected for running the model is located in the North Adriatic Sea, along the coastline of Emilia-Romagna region, where mussel farming represents an important activity (21.6 10oduction ), and aqoduction . Model aIn order to apply the model, environmental data were extracted from NetCDF (Network Common Data Form) datasets, and converted in data matrices . Hence, the forcing variables time series at each grid cell were used as input for the RAC model functions. RAC outputs were stored in a matrix, and associated to the cell spatial coordinates. Two maps of the number of days required to reach both sizes for commercialization were finally created, and the complete set of RAC daily outputs were stored in NetCDF files. R code to run the individual mussel model over a spatial domain is reported in https://esgf-index1.ceda.ac.uk/projects/esgf-ceda/) taking advantage of the results obtained within the EURO-CORDEX initiative ([Time series of future SST data were downloaded from the CEDA ESGF data node (tiative ( Coordinatiative  scenari-1), and the sum of faeces produced and uneaten feed per individual seabream (g d-1). For mussels, predicted values are in agreement with the measurements performed in eastern Canada [-1 d-1, and for seabream simulations agree with the results published for the Southern Adriatic Sea [-1 d-1.n Canada , with faatic Sea , with vaAs one can see in e.g. integrating the outputs obtained from this tool with site selection criteria  in a multi-criteria evaluation process [This paper provides a synthetic overview of design and applicability of the RAC package. The reasonableness of RAC estimations was confirmed by the agreement between the results obtained in test cases performed within the present work with previous studies in which models were validated. Runs carried out demonstrate the potential of RAC for integrating different open products nowadays provided by remote sensing and operational oceanography. This goes in the direction of overcoming limitations imposed by data scarcity, recognized as a major obstacle to the application of virtual technologies for aquaculture . Indeed, process , and con process . RAC out process , in orde process .S1 FileTable A. Model state variables, forcings, and functional relationships of M. galloprovincialis–as in Brigolin et al. (2009).Table B. Functional expressions used in the individual growth models of D. labrax and S. aurata–as in Brigolin et al. .Table C. Model state variables, forcings, and functional relationships of R. philippinarum—as in Solidoro et al. (2000).(PDF)Click here for additional data file.S2 FileTable A. Parameters used in the Mytilus galloprovincialis growth model.Table B. Parameters used in the Sparus aurata (SA) and Dicentrarchus labrax (DL) growth models.Table C. Parameters used in the Ruditapes philippinarum growth model.(PDF)Click here for additional data file.S1 Appendix(PDF)Click here for additional data file.


P value obtained at 0.001  and 0.209 . In conclusion, there are risk factors such as age and occupation that have a correlation of being infected with hepatitis C serological status.Hepatitis is an inflammation of the liver commonly caused by viral infection such as hepatitis A, B, C, D, and E but it is also possible by other causes. Infection with hepatitis C virus is also referred to as a disguise because the early infection is often asymptomatic that often goes undetected. This study aims at determining the several associated risk factors with hepatitis C serological status. The study design is cross-sectional. The biomedical data collection was carried out in 33 provinces in Indonesia with a population in urban blocks, census in Indonesia, where the sample is all household members over the age of one year from selected households by signing the informed consent. Total block census in selected urban area is about 971-block census with a total sample of 15.536 households. The results showed that there is a correlation between hepatitis C serological status and demographic group and that the age and occupation groups showed significant  Hepatitis is an inflammation of the liver that can be caused by bacteria, viruses, autoimmune processes, drugs, fatty liver, alcohol, and other harmful substances. The most potential cause of hepatitis is an infection caused by viral hepatitis A, B, C, D, and E. Hepatitis virus is the cause of burden disease and death and it has potential to become an outbreak and a high risk of disease transmission. Hepatitis C typically only shows light such as tiredness, fever, nausea or poor appetite, muscle aches, joint pain, and pain in the liver area. Hepatitis C infection is also known as the silent infection since, in early infection, it is often asymptomatic with no typical symptoms and so it is often overlooked. This condition may continuously progress into an extensive liver damage, which is known as cirrhosis of liver. In some cases, cirrhosis will develop into liver failure, including liver cancer. Most people do not know that they are infected with hepatitis C till the liver damage appears, or through blood tests in the laboratory , 2.At the prior infection, only about 25% of patients show acute hepatitis symptoms, such as weakness, muscle pain, loss of appetite, and fever or sometimes it appears jaundiced on skin and eyes (jaundice), about 75% of patients are without symptoms, even the disease becomes chronic, and often the patient remained asymptomatic (without symptoms). Approximately, 10–15 percent of patients will heal themselves without treatment. This infection is commonly suffered by young women rather than other groups –5.It is estimated that about 50% of hepatitis C cases is transmitted through the use of drugs that are injected . People About 5–50% infected people in the United States and Canada know their status. The examination of hepatitis C is recommended for people at high risk, including those who have tattoos . ScreeniThe diagnosis of hepatitis C is found if there is HCV RNA in the serum of patients. The tests are very expensive. Therefore, the examination of anti-HCV by ELISA method can be used to screen patients whom are suspected of having hepatitis C, due to its low costs. If the result of anti-HCV shows positive, then it needs to be confirmed by HCV RNA examination. The HCV genotype examination should also be carried out for all patients who will get antiviral therapy to assess the duration of treatment required and the expected response to therapy .Both hepatitis C and hepatitis B are similarly transmitted through blood. In 2007, Ministry of Health Republic of Indonesia conducted National Basic Health Research. The data was collected using a structured questionnaire and also laboratory examination for several diseases including antibody testing of hepatitis B and C. The aim of this study is to determine the association of several risk factors with the serological status of hepatitis C. The analysis of risk factors such as age, gender, medical history, injury/accident history, dental care, occupation, thalassemia, hemophilia, and blood transfusion was conducted based on a questionnaire and linked with the data of anti-hepatitis C serological status.In 2007, National Institute of Health Research and Development (NIHRD), Ministry of Health Republic of Indonesia, had conducted the first National Basic Health Research, a nationwide survey with cross-sectional design. Samples of National Basic Health Research 2007 followed the framework of selected National Survey on Social and Economy in 2007. The National Basic Health Research could describe the health profile of the district/city or province level. From each district/city included in the sample frame district/city taken, a number of census blocks were proportional to the number of households in the district/city. A census block included in the sample census block on a district/city is proportional to the number of households in a district/city . Overall, a sample census block of the National Survey on Social and Economy 2007 was selected from 17,357 census block samples. In 2007, the National Basic Health Research was able to visit 17,150 census blocks of 438 districts/municipalities. In addition, beside the public health data collection, the survey was also gathering the biomedical data with specimens collection and laboratory testing [Biomedical data was conducted in 33 provinces in Indonesia with the population in urban census blocks in Indonesia. Blood sampling was also conducted on all household members over the age of one year from selected households with a signed informed consent. The series of biomedical sampling selected about 15% of total urban census block of National Survey on Social and Economy. The number of selected census blocks in urban areas reached 971 blocks selected with a total sample of 15,536 households. Blood sampling was not conducted on household members who have had bleeding background regularly.P < 0.05 was taken as significant for interpretation.Statistical analysis was performed with SPSS 15 software. Chi-square test was applied for analysis of categorical data. A P = 0.001 by Odds Ratio (OR) = 3.27 and 95% Confidence Interval (CI) = 1.84–5.81. Occupation shows a correlation with hepatitis C serological status.In Hepatitis C is an infection which attacks the liver. One of the causes of hepatitis C is the hepatitis C virus (HCV). Hepatitis C is often asymptomatic, but chronic infection can cause scarring of the liver and lead to cirrhosis. In some cases, people who have cirrhosis also suffer liver failure, liver cancer, or swollen veins in the esophagus and stomach, which can cause bleeding into death . Anti-HCHepatitis C showed acute symptoms only in 15% cases . These iEighty percent of persons exposed to hepatitis C will develop chronic infection . Most peHepatitis C is transmitted through blood and medical equipment contaminated with hepatitis C. According to previous numerous literature, several transmission processes of hepatitis C in the world are caused by the use of injecting drug (60%), through sexual relations 15%), with blood transfusions before screening making up about 10% and the use of means of hemodialysis and perinatal care occupying about 5%. Hepatitis C in developed countries is transmitted through injecting drug use (IDU), whereas, in developing countries, it is transmitted through blood transfusions and unsafe medical equipment . Accordi%, with bP = 0.050 and risk factors (OR) about 2,26 and 95% Confidence Interval (CI) about 0.98–5.2. Like the research conducted in Egypt, hepatitis C in this country is transmitted through unsterilized surgical equipment in dental and general health services [P = 0.001. Since the Odd Ratio result is less than number one, then the relation is not a risk factor but protective one. While on a variable, respondents who do not work have relation with serological status hepatitis C with the result and OR = 0.23, 95% CI (0.594 to 0.129), and P = 0.209. The effect is protective because the OR result is less than one. Works related to medical devices contaminated with hepatitis C virus can transmit to workers related to the equipment. Hospital equipment can also transmit hepatitis C virus, including the use of needles and syringes, drugs vials that are continuously used, infuse bags, and unsterilized surgical equipment [In this study, we also found a correlation between the variables of people who have oral and dental care at a health facility in the last 12 months; they are going to obtain results with no significance with services . Likewisquipment , 16.Several risk factors such as dental treatment, thalassemia, and blood transfusion had no correlation of being infected with hepatitis C. The age group 5 years has a high risk to be infected with hepatitis C.


Studies suggest that brain-derived neurotrophic factor (BDNF) and the hypothalamic-pituitary-adrenal (HPA) axis modulate dopaminergic activity in response to nicotine and that the concentrations of BDNF and cortisol seem to be dependent on the amount and duration of smoking. Therefore, we investigated BDNF and cortisol levels in smokers ranked by daily cigarette consumption. Twenty-seven adult males (13 non-smokers and 14 smokers) participated in the study. The smokers were divided in two groups: light (n=7) and heavy smokers (n=7). Anthropometric parameters and age were paired between the groups, and plasma BDNF and salivary cortisol levels were measured. Saliva samples were collected on awakening, 30 min after awakening, at 10:00 and 12:00 am, 5:00 and 10:00 pm. Additionally, cotinine serum levels were measured in smokers. Heavy smokers had higher mean values of BDNF compared to the control group (P=0.01), whereas no difference was observed in light smokers. Moreover, heavy smokers presented lower cortisol levels in the last collection (10:00 pm) than the control group (P=0.02) and presented statically higher values of cotinine than the light smokers (P=0.002). In conclusion, changes in BDNF and cortisol levels (10:00 pm) appear to be dependent on heavy cigarette smoking and can be involved in activation and in the relationship between the mesolimbic system and the HPA axis. Cigarette smoke is a mixture of gases and particles, and nicotine is the main addictive substance in tobacco. After each inhalation, nicotine rapidly reaches the brain, where it acts primarily on dopaminergic neurons in the mesolimbic system (reward system), leading to dopamine release, which is responsible for euphoria and pleasure feelings and constitutes the key component of addiction .Brain-derived neurotrophic factor (BDNF) is a key molecule related to the modulation of this system. BDNF is one of the most abundant neurotrophins in the brain and it is involved in neuronal growth, differentiation and survival. Moreover, BDNF has been identified as a regulator of synaptic plasticity , which iIn addition to BDNF, studies have demonstrated that the hypothalamic-pituitary-adrenal (HPA) axis also seems to mediate dopaminergic activity ,7, influTherefore, because both BDNF and the HPA axis appear to modulate dopaminergic activity in response to nicotine exposure, this study simultaneously evaluated central (BDNF) and peripheral (cortisol) components related to nicotine consumption by smokers. Furthermore, since the levels of both BDNF and cortisol in smokers seems to be dependent on the amount and duration of smoking, we proposed to investigate BDNF and cortisol levels in healthy smokers, stratified by daily cigarette consumption; previous studies have not yet been carried out. Therefore, this pilot study aimed to evaluate the BDNF and cortisol levels in healthy adult smokers, ranked by daily cigarette consumption and compared to healthy non-smokers.2); not currently using anti-inflammatory medications; self-reported absence of cough, infection, fever and flu in the month prior to the assessments; and no use of nicotine replacement therapy in the last three months. Control subjects could not have been passive smokers and cigarettes used by smokers must have been manufactured with a filter. The exclusion criteria were the lack of normal circadian rhythm: salivary cortisol at 10:00 pm <6.0 nmol/L and an 8:00 am/10:00 pm ratio of salivary cortisol >2. This study was performed according to Resolution No. 466/12 of the National Health Council. The Research Ethics Committee of the Universidade Federal dos Vales do Jequitinhonha e Mucuri, Brazil, approved this study (protocol No. 003/12). All participants gave written informed consent.Thirty-five healthy adult men (17 non-smokers and 18 smokers), aged 18–45 years, were recruited by personal invitation during visits to homes and health centers in the local community. To be included in the study, the subjects had to meet the following criteria: normal lung function; self-report of no current acute or chronic diseases; be eutrophic according to the body mass index , forced vital capacity (FVC) and FEV1/FVC were calculated in accordance with the American Thoracic Society/European Respiratory Society . The forced expiratory volume in 1 s , the samples were stored at –80°C until further analysis.One week after the clinical assessment, the subjects underwent cortisol evaluation. Because the cortisol level increases 30 min after awakening and declines throughout the day, multiple measurements of cortisol levels during the day provide a more valid information regarding the daily cortisol release and reflect cortisol circadian rhythm. Saliva samples were, therefore, collected upon awakening and after 30 min, at 10:00 and 12:00 am, and 5:00 and 10:00 pm. For this purpose, the participants were instructed to collect saliva samples using a Salivette¯ . Volunteers were instructed not to smoke before the first collection (immediately upon awakening) and to avoid smoking for at least 30 min before each collection throughout the day. In addition, they were instructed to avoid alcoholic beverages, physical activity and eating 30 min before sample collection and brushing their teeth 2 h before sample collection. All collections were performed in the subject's house. After each collection, the subjects stored the Salivettes at –20°C. On the following day, the researchers transported the samples under refrigeration to the laboratory, and after centrifugation  were used for cortisol analyses. The procedures were performed according to the manufacturer's specifications. The test has a low sensitivity limit of 0.19 nmol/L. In the present study, the sensitivity ranged from 0.33 to 82.77 nmol/L, and the average intra-assay and inter-assay coefficients of variation were 3.3 and 4.3%, respectively.The day after cortisol evaluation, blood was collected between 6:00 and 8:00 am for BDNF analysis in all subjects and for cotinine analysis in smokers. The blood was collected aseptically by puncturing the median cubital vein, after 8–12 h of fasting and abstaining from cigarettes. The BDNF plasma levels were measured with an ELISA kit , according to the manufacturer's instructions. The limit of detection was 5 pg/mL.To assess the exposure to cigarette smoke and the amount of nicotine absorbed, levels of serum cotinine were measured in smokers. This analysis was performed in a privately-owned laboratory using a chemiluminescence method. A level of cotinine higher than 25 ng/mL was considered the reference value for smokers.post hoc test. Comparison of the smoking history variables between light and heavy smokers was performed with the independent t-test. The level of statistical significance was P≤0.05, and data are reported as mean±SD.Statistical analyses were performed using the GraphPad Prism 5  statistical package. The normality of data was checked by the Shapiro-Wilk test and the homogeneity by the Levene test. As the data were normally distributed, the comparison of results of the light and heavy smokers and the control group was performed by ANOVA and the Scheffé 1, FVC, FEV1/FVC, pack-years and smoking time (P>0.05). In relation to the level of cigarette exposure, heavy smokers presented statistically higher cotinine levels than light smokers (P=0.002).Twenty-seven subjects (13 non-smokers and 14 smokers) completed the study. Four subjects from each group were excluded for not having a normal circadian rhythm. All the light smokers consumed 10 cigarettes/day, and all heavy smokers consumed 20 cigarettes/day. The general characteristics and smoking history of the participants are shown in Plasma BDNF levels are presented in To our knowledge, this is the first study to simultaneously assess BDNF and cortisol levels in healthy smokers. Our data demonstrate that heavy smokers presented an increase in BDNF and a reduction in cortisol levels measured at 10:00 pm compared to non-smokers. These results corroborate other studies in humans that demonstrate a dependence between BDNF and cortisol levels that is related to the amount of smoking ,8.Pre-clinical data have shown that nicotine induces BDNF release and upregulates BDNF receptors in the rat brain . In humaSimilarly to BDNF, the cortisol response to nicotine seems to be dependent on the frequency of exposure . RecentlIn addition, the lower cortisol level in heavy smokers could be caused by the stimulation of the mesolimbic system because the activation of the reward circuitry underlying addictive behavior results in the release of excitatory neurotransmitters in brain areas, such as the hippocampus and amygdala . AlthougAs mentioned above, cortisol modulates dopaminergic activity in the reward system, possibly caused by a negative feedback, because cortisol increases dopamine levels and dopamine receptor stimulation promotes a decrease in cortisol . StrengtMoreover, although light smokers did not differ from control subjects in the variables assessed, such as BDNF and cortisol, an increase in cigarette consumption could lead to a pattern similar to that of heavy smokers and promote a higher risk for smoking-related diseases. It is believed that plasma nicotine levels are directly influenced by the number of cigarettes smoked per day. This hypothesis was confirmed by the significant difference in cotinine levels in heavy and light smokers. Furthermore, considering that higher BDNF levels were also observed in heavy smokers, it seems that the increase in nicotine exposure could have increased the activation of the mesolimbic system, thereby modifying the dopaminergic activity and decreasing cortisol levels.This study has limitations, and the results must be interpreted within the context of its design. Although BDNF was evaluated in plasma, studies have demonstrated that peripheral blood can provide an important reservoir of BDNF. In this context, the BDNF content evaluated in serum could be directly influenced by nicotine-lung stimulation because the smooth muscle of the airway is a significant source of BDNF in response to oxidant stress induced by cigarette smoke . MoreoveAlthough the present study needs to be confirmed in a large random series of smokers and non-smokers, the consistency of the results observed supports our conclusions. Furthermore, the results of this study provide subsidies for understanding the effect of nicotine consumption on BDNF and cortisol levels in healthy smokers. In summary, levels of BDNF and cortisol measured at 10:00 pm appear to be dependent on cigarette smoking and could represent the relationship between the mesolimbic system and the HPA axis in the mechanism of nicotine addiction. These results bring new perspectives for the investigation of this mechanism and, consequently, of pharmacological interventions.


Knowledge of the lung vessel morphology in healthy subjects is necessary to improve our understanding about the functional network of the lung and to recognize pathologic deviations beyond the normal inter-subject variation. Established values of normal lung morphology have been derived from necropsy material of only very few subjects. In order to determine morphologic readouts from a large number of healthy subjects, computed tomography pulmonary angiography (CTPA) datasets, negative for pulmonary embolism, and other thoracic pathologies, were analyzed using a fully-automatic, in-house developed artery/vein separation algorithm. The number, volume, and tortuosity of the vessels in a diameter range between 2 and 10 mm were determined. Visual inspection of all datasets was used to exclude subjects with poor image quality or inadequate artery/vein separation from the analysis. Validation of the algorithm was performed manually by a radiologist on randomly selected subjects. In 123 subjects (men/women: 55/68), aged 59 ± 17 years, the median overlap between visual inspection and fully-automatic segmentation was 94.6% (69.2–99.9%). The median number of vessel segments in the ranges of 8–10, 6–8, 4–6, and 2–4 mm diameter was 9, 34, 134, and 797, respectively. Number of vessel segments divided by the subject's lung volume was 206 vessels/L with arteries and veins contributing almost equally. In women this vessel density was about 15% higher than in men. Median arterial and venous volumes were 1.52 and 1.54% of the lung volume, respectively. Tortuosity was best described with the sum-of-angles metric and was 142.1 rad/m (138.3–144.5 rad/m). In conclusion, our fully-automatic artery/vein separation algorithm provided reliable measures of pulmonary arteries and veins with respect to age and gender. There was a large variation between subjects in all readouts. No relevant dependence on age, gender, or vessel type was observed. These data may provide reference values for morphometric analysis of lung vessels. TRadiologic evaluation of lung vasculature in thoracic computed tomography (CT) images allows the description of dilatation, tortuosity, and tapering of the pulmonary vessels down to diameters of about 1 mm . The anonymized datasets were transferred to an independent workstation. The images from both detectors (tin-filtered 140 and 100 kVp) were equally mixed and used for the analysis. In addition to the CT datasets, only information on age and gender was available for each subject.The CTPA images were acquired with a 128-slice dual-source CT scanner . The detailed description of the imaging protocol is presented in Meinel et al. . BrieflyThe automatic vessel segmentation was performed by the in-house developed software as shown in Figure musculus erector spinae was determined in order to quantify the influence of image quality on the readouts. Likewise, the mean attenuation in the main pulmonary artery was measured to evaluate the influence of the contrast material concentration in the lung vasculature on our readouts.Detailed evaluation of the artery/vein separation algorithm showed that in few subjects the algorithm may wrongly assign all arteries and veins in whole lung lobes  and the distance metric (DM)  or mean ± standard deviation, if they failed or passed the normality test, respectively, unless otherwise stated. P-values (p) < 0.05 were considered statistically significant.Statistical analysis was performed in SPSS  and GraphPad Prism . Agreement between visual inspection and the manual labeling of the artery/vein separation was analyzed using Bland-Altman analysis. The impact of gender, age, mean X-ray attenuation and vessel type (artery/vein) on artery/vein separation was analyzed using a General Linear Model (GLM). Therefore, the mentioned variables and the interaction gender*age were included in the analysis. Other readouts were tested for normality with the D'Agostino and Pearson omnibus normality test. Differences between men and women in these readouts were evaluated by One hundred and thirty six subjects (men/women = 64/72) were included in this retrospective study. For each subject information on age and gender was available. CTPA examinations resulted in a mean dose-length product of 299 mGycm (230–367 mGycm) corresponding to a mean effective radiation dose of 5.7 mSv . Further, the distribution of the DM values was very close to the minimum of 0%, which makes it hard to detect changes at this side of the distribution. Therefore, we have chosen to measure tortuosity with SOAM in this study. For comparison with other studies we present the results for DM in Supplementary Table Figure p < 0.001). Standard deviation of the X-ray attenuation in the musculus erector spinae was 25 HU (19–32 HU) without a significant difference between men and women. X-ray attenuation in the main pulmonary artery was 351 HU (300–444 HU). Attenuation in women was higher than in men . The algorithm detected 977 vessel segments per subject  in the diameter range of 2–10 mm. These were labeled as 487 arterial segments (390–587 segments) and 471 venous segments (371–551 segments). Representative CT images and a 3D rendering of the labeled vessels are presented in Figures Mean age of the subjects was 59 ± 17 years with no significant difference between men and women. The mean lung volume determined from the CTPA images was 4.6 ± 1.3 L and was significantly larger in men than in women  vs. 176/L (152–209/L), respectively; p < 0.001]. There was no significant offset associated with gender for normalized vessel volume. Tortuosity of all vessels was 140.8 rad/m (137.6–144.5 rad/m) in men and 142.7 rad/m (138.8–144.8 rad/m) in women and did not show a significant offset.The number of vessel segments in the diameter ranges of 8–10, 6–8, 4–6, and 2–4 mm were 9 (6–12), 34 (25–44), 134 (102–159), and 797 (623–905), respectively . Despite these differences, the strong influence of age and X-ray attenuation in the truncus pulmonalis on the respective values for arteries resulted in a virtually complete overlap between arteries and veins  and 471 veins (371–551 veins). Vessel density was just above 100/L for both vessel types (Table p = 0.03) and a trend concerning normalized vessel volume . In this readout, arteries but not veins showed a slight increase with age, although this increase was not significant by itself. The increase in tortuosity with age for arteries and veins was not significant.In women, the number of all vessel segments increased with age by 2.4 vessels/year, the vessel density by 0.49/L/year, and the normalized vessel volume by 0.004%/year . In both readouts, arteries showed a significantly stronger increase with attenuation compared to veins . In the normalized vessel volume, we observed a weak, non-significant decrease with higher attenuation. With increasing attenuation slightly more tortuous vessels were detected (p = 0.001).Increasing X-ray attenuation in the ty Table . HoweverMorphometric information on the lung structures in healthy subjects is necessary to improve our understanding about the complex and interconnected functional network of the lung Weibel, , 2017. TWe found that the number of detected vessels was strongly related to the lung volume. The normalized number of vessels larger than 2 mm in diameter was about 200 per liter lung volume. The number comprised about 100 arteries/L and 100 veins/L. These numbers were independent of age; however, they were about 15% higher in women than in men. This may have resulted, in part, from the higher pulmonary artery attenuation in women observed in the analyzed datasets. Despite the higher vessel density in women, the larger lung volumes in men resulted in more vessel segments over all diameter ranges. Since the anatomy of the lobar and segmental lung vessels is the same for men and women, the smaller vessels in women are subsequently counted in smaller diameter ranges than the anatomically comparable vessels in men. This shift can explain the lower number of vessel segments in women.A direct comparison to previous studies on vessel casts is only possible for the study from Huang et al., which also presents a connection matrix converting Strahler orders to vessel segments (Huang et al., The total volume of detected vessels accounted for about 3% of lung volume. This was independent of age and gender. Arteries and veins contributed to this volume by about 50% each. Notably, there was a trend toward an age-dependent decrease of the venous volume in men which might be due to age-related emphysema. Unfortunately we had no lung function data to substantiate this speculation.To our knowledge, our study is the first to quantify the tortuosity of both arteries and veins by age and gender. We used both distance metric and SOAM to examine vessel tortuosity. Although DM has been more widely used, we found SOAM more adequate as a measure because its distribution compares to a Gaussian distribution, whereas DM is very much skewed. SOAM was about 142 rad/m, independent of age and gender in both arteries and veins. This corresponds to an angle of approximately 9° between successive 1 mm long sections along the path of the vessel segments. It should be noted that SOAM depends on the length of the individual sections used. First, longer sections would average out small kinks and turns in the vessel paths, resulting in smaller values for the metric. And second, the measure itself depends on the length of the sections. Therefore, comparisons to other datasets should account for the section length used in the analysis.musculus erector spinae as a measure to assess the influence of image quality on the results. Both, number of vessel segments, and vessel density increased with increasing noise, which was driven by a small number of subjects (see Supplementary Figure Prior to inclusion into this study, all CTPA scans were examined for artifacts or inadequate vascular enhancement. Therefore, one can assume that the clear majority of vessels are depicted correctly. Still, image noise can be a source of error in the segmentation. With increasing noise, the measure identifying tube-like structures in the CTPA images becomes weaker and the algorithm might miss small vessels. We used the standard deviation of the X-ray attenuation of the truncus pulmonalis. The large variation remained even when the mixed model was used to correct for these factors (Supplementary Figure All readouts showed a large variation between the individual subjects independent of age, gender, vessel type, and X-ray attenuation in the truncus pulmonalis and veins running into the left atrium is subject to future development. While the algorithm runs fully automatically, the inadequate artery/vein labeling in approximately 10% of subjects necessitates a critical inspection of the results. Regarding the current state of the art, we consider this necessary for any algorithm. We only had information on age and gender available, thus we cannot exclude influences of other parameters on our morphological readouts. This includes the degree of inspiration which influences the lung volume and, therefore, the vessel density, relative vessel volume, and likely also the vessel tortuosity. However, under the assumption that this variation is normally distributed within our large sample, we can expect that this does not have an influence on our results, but rather reflects the variation expected in clinical routine. Finally, due to radiation exposure one cannot repeatedly scan the patients. This would be necessary to determine the reproducibility of the readouts in the same subject.We took advantage of a series of CT investigations performed in a standardized way in a high-volume diagnostic center. All patients were free of acute or chronic pulmonary embolism or other abnormalities of the heart or the lung that could be detected by CT. Unfortunately we had no information on co-morbidities like lung function abnormalities, arterial hypertension, diabetes, non-thoracic cancer, etc. Therefore, we cannot exclude that such age-related factors may have influenced our morphometric results. However, such a group of patients may better represent subjects undergoing thoracic CT investigations than a highly selected population of absolutely healthy subjects. Thus, this study provides normal ranges of the morphologic readouts applicable to the identification of patients in CAD algorithms. A further limitation of this study is the limited number of vessels accessible by CT imaging and hence by the algorithm. A typical human lung includes hundreds of millions of vessels. E.g., Huang et al. found a total of 15 generations of vessels between the main pulmonary artery and the capillaries, with diameters varying from 15 to 0.02 mm (Huang et al., We report quantitative readouts of lung vessel morphology from a large cohort of subjects with no apparent thoracic abnormalities examined by means of thoracic computed tomography and analyzed by means of a fully-automatic artery/vein separation algorithm. We obtained normal values of lung vessel morphology and provide information on their variation with respect to age and gender. These data may serve as reference values for morphometric analysis and may be useful for the identification of pathologic changes in chronic lung diseases.MP: conceived the study, acquired data, designed and executed the experiments, analyzed and interpreted the data, wrote and revised the manuscript; CB, CP, and AA: executed the experiments, analyzed and interpreted the data, read and revised the manuscript; MU, RS, and AO: guided the experimental design, read and revised the manuscript; HO: guided the experimental design, analyzed and interpreted the data, read and revised the manuscript; TJ: conceived the study, acquired data, read and revised the manuscript; FM: conceived the study, acquired data, guided the experimental design, read and revised the manuscript; ZB: conceived the study, guided the experimental design, analyzed, and interpreted the data, wrote and revised the manuscript. All authors gave final approval of the manuscript to be published.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


After the intervention, hemoglobin (p < 0.05) and hematocrit (p < 0.05) levels were significantly improved, thereby almost halving the rate of anemia. These changes were significantly larger in the baseline anemic group than the non-anemic group (p < 0.01). BMI significantly increased in the baseline underweight/normal group (p < 0.001) but not in the overweight/obese group. The school lunch intervention significantly improved nutritional intakes and health statuses, implying its potential for reducing anemia and resolving the double burden of malnutrition among rural Indonesian schoolchildren.School lunch is not provided in public elementary schools in Indonesia, and students frequently buy and eat snacks at school. We hypothesized that providing a traditional Sundanese meal as school lunch would be beneficial for children in rural West Java. To test this hypothesis, we evaluated the effect of a 1-month school lunch intervention aiming at sustainability and based on children’s nutritional intake, hemoglobin and hematocrit levels, and body mass index (BMI). A lunch  containing one-third of the recommended daily allowance of energy was offered every school day for 1 month, targeting 68 fourth-grade elementary schoolchildren. At baseline, the prevalence of anemia was 33.3%. The prevalence of stunting and underweight were 32.4% and 2.9%, respectively, whereas that of overweight and obesity combined was 17.6%, indicating a double burden of malnutrition among the subjects. During the intervention, intakes of protein ( In this program, a midmorning snack was provided 3 days/week throughout the year or 108 snacks in total per year; each snack contained a minimum of 300 kcal energy and 5 g protein, and the allocated cost for each snack was approximately US $0.10–0.15. The program was expanded to all provinces in 1998, targeting 8.1 million children in 53,000 schools. However, because of the severe Asian economic crisis around 1998, it was soon canceled [n = 68) than in non-snacks (n = 153). The ratios of the nutrient density in snacks to that in non-snacks were 1.31 for fat, 1.09 for carbohydrate, 0.54 for protein, 0.43 for iron, 0.25 for phosphorus, 0.07 for calcium, 0.03 for vitamin A, and 0.02 for vitamin C. Furthermore, children’s expenditure on snacks constituted a heavy financial burden for their parents [We conducted nutrition and health surveys in the target village located in West Java, particularly in children. One of the most important findings was that elementary school children frequently bought and ate snacks not only before and after school hours but also during recess in school; such snacking played a crucial role in worsening their nutritional status. In addition, their nutritional status deteriorated simply because the snacks were of very poor nutritional quality . Our ana parents . To imprIn this study, we conducted a 1-month school lunch intervention in a locally sustainable manner, involving the provision of locally available food cooked by villagers, with the advice of nutritionists. Moreover, we considered that the total per-day per-person cost for providing lunch should be equivalent to the per-day per-person expenditure on snacks. This study aimed to clarify the effect of the intervention trial on children’s nutritional status, hemoglobin and hematocrit levels, and BMI.The study village Sukajadi is located in the kecamatan (subdistrict) Tamansari, kabupaten (district) Bogor, West Java, Indonesia. Detailed information regarding the study area is reported elsewhere ,12. We tAll 68 subjects were students in the fourth grade at the target elementary school. There were two reasons for selecting fourth-grade students as follows: (1) their adequate memory of the food they had eaten in the past 24 h, and (2), they were in the growth stage before the onset of puberty. Written informed consent was obtained from the parents of each child after explaining the purpose and procedures of the study. The study protocol was approved by the Ethics Committee of the Graduate School of Frontier Sciences, The University of Tokyo.The intervention of providing lunch was performed on every school day for 1 month from August to September 2015. To assess the effect of this intervention, baseline and follow-up surveys were conducted one day before and after the intervention.Given that no school feeding trials had been previously performed in the study area, there was no kitchen or cook at the school. In this study, three local health assistants  participated as cooks and the kitchen at the home of one Kader was used for preparing the dishes. Three Kader started cooking at 05:00 and finished around 09:30 and then packed the cooked food in plastic lunchboxes around 11:00 to be carried to the school . The chiThe menu for the lunch each day for 1 month was planned by one of the authors (KR), following two principles. One was derived from the nutritional rationality of providing one-third of the energy required per day, based on the Japanese school lunch guidelines by the Ministry of Education, Culture, Sports, Science and Technology . The othBecause the local tube well was temporarily broken during the intervention period, the students used a water tank, through which spring water was run using a bamboo pipe, for washing their hands. Two nutritionists who were stationed at the school every day instructed the children to wash their hands before eating, to pray just before eating, and to finish the meal within 30 min. The nutritionists also weighed and recorded the leftovers, if any, in each child’s lunchbox.Intensive data collection was performed twice: in the baseline survey performed 1 day before the intervention and in the follow-up survey performed 1 day after the intervention. In both surveys, we conducted anthropometric measurements, a dietary intake survey, and blood sampling and conducted a questionnaire survey regarding children’s food-related knowledge, behavioral patterns, and socioeconomic features of their households, the data of which are not dealt with in detail in this study. The student’s mother (or caretaker in a few cases) was also invited to attend both baseline and follow-up surveys to assist the child with the dietary intake survey and the questionnaire on socioeconomic features of households.z-scores  and BMI-for-age (BMIZ)) were calculated based on the WHO growth references published in 2007 [z-scores, namely HAZ and BMIZ, was performed based on their month-based ages.Anthropometric measurements were conducted by one of the authors (MS), with the assistance of an Indonesian nutritionist. Height was measured to the nearest 1 mm using an anthropometer and body weight was measured with minimum clothing to the nearest 0.1 kg using a Tanita digital weighing scale . Mid-upper arm circumference was measured to the nearest 1 mm using a plastic measure tape. Skinfold thickness at the biceps, triceps, and subscapular area were recorded to the nearest 0.1 mm using Eiken calipers . From each child’s height and weight, two  in 2007 . The ageChildren’s dietary intakes in terms of energy and nutrients were assessed using a 24 h recall method, which was asked to both the child and his/her mother (or caretaker). To guarantee the accuracy of the reported amounts of food consumed as estimated in this 24 h recall procedure, we performed the following steps. First, all snack food available to children at local stores or stalls in and around the schoolyard was weighed. Second, all food (except that in cooked dishes) consumed at home was weighed for various sizes of each foodstuff to determine the “portion size” for each child ; for food in cooked dishes, we used the “standardized” recipe data based on our previous study in this village  to estimA non-fasting 2.0 mL sample of venous blood was obtained from each child by qualified medical staff. Immediately after collection, a small portion was used to measure hemoglobin and hematocrit levels. The remaining portion was aliquoted into a heparinized tube, stored at 4 °C, and centrifuged for 30 min on site. The obtained plasma samples were stored at −20 °C until hormone analysis (equipment for storing samples at −80 °C was unavailable). Capillary tubes, filled with fresh blood and stoppered in the field, were taken to a laboratory at the Bogor Agricultural University. These samples were then centrifuged to determine the hematocrit level. Using 20 μL of blood, hemoglobin levels were determined by the cyanmethemoglobin method. For both hemoglobin and hematocrit, the cut-off values suggested by WHO  were uset-test , or Mann–Whitney U test . Energy and nutrient intake and hemoglobin and hematocrit levels before and after the intervention were compared using paired t-test  or Wilcoxon signed-rank test  and the anemic status was compared using the chi-square test. Hemoglobin and hematocrit levels were compared using two-way analysis of variance (ANOVA) with repeated measures on two factors, namely ‘baseline anemic status’ (two levels: anemia or non-anemia) and ‘time’ (two levels: before and after the intervention), together with between-factors interaction term. Similarly, BMI and BMIZ levels were compared using two-way ANOVA with repeated measures on two factors, namely ‘baseline BMIZ status’ (two levels: BMIZ < 1 and BMIZ ≥ 1) and ‘time’ (two levels: before and after the intervention), together with between-factors interaction term. Changes in hemoglobin  and hematocrit  levels before and after the intervention were compared between the anemic and non-anemic groups, as determined in the baseline survey. Statistical significance was defined as p < 0.05. All analyses were performed using the Statistical Package for Social Science software package .The normality of the distribution of data was examined using the Kolmogorov-Smirnov test. Characteristics of the children were compared between boys and girls using the chi-square test , t-test  or Wilcoxon signed-rank test  revealed that the intake of protein, calcium, and vitamin C significantly increased, that of fat significantly decreased, and that of energy and other nutrients, namely carbohydrate, phosphorus, iron, and vitamin A, did not significantly change. When compared with the Indonesian RDA, children’s NAR before and during the intervention correspondingly differed, with the same statistical differences as in the amount of intake. Larger NARs of protein, calcium, and vitamin C during the intervention were of particular interest because they were judged to be less before the intervention, despite their significant roles in human health, particularly healthy growth.n = 51 because of failure to sample blood for 17 of them). Both hemoglobin  and hematocrit  levels were significantly improved after the intervention. Hemoglobin and hematocrit levels were compared using two-way ANOVA with repeated measures, with ‘baseline anemic status’ (two levels: anemia or non-anemia) and ‘time’ (two levels: before and after the intervention) as the independent valuables. The results showed that there was a significant interaction between baseline anemic status and time . There were also main effects of baseline anemic status  and time . Anemia prevalence was 33.3% at baseline, exceeding the national average (26.4%) [p < 0.01); in the anemic group, hemoglobin levels improved in all individuals, while hematocrit levels improved in 82% of them.n = 54) and high-BMI groups  based on the measurements before the intervention or baseline. BMI and BMIZ levels were compared using two-way ANOVA with repeated measures, with ‘baseline BMIZ status’ (two levels: BMIZ < 1 and BMIZ ≥ 1) and ‘time’ (two levels: before and after the intervention) as the independent valuables. The results revealed that there was no interaction between baseline BMIZ status and time . There was no main effect of time , however, there was a significant main effect of baseline BMIZ status . In the baseline underweight/normal or low-BMI group, both BMI (p < 0.001) and BMIZ (p < 0.05) were significantly higher after the intervention than before the intervention. In contrast, in the overweight/obese or high-BMI group, neither BMI nor BMIZ showed a significant difference. Notably, BMI was significantly higher after the intervention when all subjects were pooled.This is the first trial regarding a school feeding intervention in Indonesia, targeting children in a rural village of West Java. The schoolchildren in the Sukajadi village have been investigated by the authors for >10 years with regard to their health and nutrition statuses ,12,18. SA previous review of school feeding programs in many developing countries worldwide reported four major findings . First, n = 153) and only 15.9 mg/MJ in snacks (n = 68) [Our study, based on the provision of locally available Sundanese food as the school lunch at a reasonable cost for 1 month, was designed to investigate the effects of such an intervention on the double burden of malnutrition. There were advantages and disadvantages to this approach. The advantages were the high acceptability of the project among the villagers, particularly the parents of the children, and the change in the food that the children frequently consumed, i.e., from less nutritious snacks to nutritious local food. The disadvantages included the inadequate amounts of calcium contained in local food compared with the Indonesian RDA; during the intervention, the mean per-day calcium intake from all food, including the provided lunch, was 240 mg or only 24% of NAR, despite the mean calcium content being 236 mg/MJ in local non-snacks ((n = 68) .r = 0.588, p < 0.05), indicating that the improvement of vitamin C intake had a positive effect on the hemoglobin status among children with low baseline iron stores. Fourth, there was a notable result regarding the changes of BMI in children after the 1-month intervention. Considering that 1-month increments of height and weight for 9-year-old children are typically only 0.5 cm and 0.3 kg, respectively [There were five main achievements of our intervention trial. First, there was a clear effect on children’s behavior associated with them simply not buying snacks in school during school hours, although their behavior was not a direct focus of the current study. Second, the children’s dietary quality improved during the intervention, while their energy intake did not change. Our baseline survey revealed that one-third of the energy intake was attributable to snacks and the intake of some micronutrients was lower than one-third of RDA before the intervention, indicating the poor quality of the children’s diet because of excessive snacking. The provided school lunch included fruits, vegetables, and animal and protein dishes every day, resulting in improving the protein, calcium, and vitamin C intakes. Such improvements in the diet imply advantageous effects of reducing the prevalence of underweight and anemia. Among the major sources of hemoglobin improvement, namely protein, iron, and vitamin C ,30, the ectively , it is jectively ; in contectively . ProvidiOur study had a few limitations. First, we applied a single 24 h recall method to assess children’s dietary intake, for both baseline and the follow-up surveys. This is despite the fact that several 24 h recalls for the same individual on nonconsecutive days are preferable to assure reproducibility of dietary intake . The maiDespite the above-mentioned limitations, the present study provides evidence of school lunch interventions’ efficacy in improving the nutritional and health statuses of school-aged children in Indonesia, as well as other developing countries that face the double burden of malnutrition. As the period of schooling is critical for formation of effective dietary habits, appropriate school feeding programs, in tandem with practicable local food culture, should be promoted.