Genetic Analysis Workshop 14 simulated data have been analyzed with MASC(marker association segregation chi-squares) in which we implemented a bootstrap procedure to provide the variation intervals of parameter estimates. We model here the effect of a genetic factor, S, for Kofendrerd Personality Disorder in the region of the marker C03R0281 for the Aipotu population. The goodness of fit of several genetic models with two alleles for one locus has been tested. The data are not compatible with a direct effect of a single-nucleotide polymorphism (SNP)  in the region. Therefore, we can conclude that the functional polymorphism has not been typed and is in linkage disequilibrium with the four studied SNPs. We obtained very large variation intervals both of the disease allele frequency and the degree of dominance. The uncertainty of the model parameters can be explained first, by the method used, which models marginal effects when the disease is due to complex interactions, second, by the presence of different sub-criteria used for the diagnosis that are not determined by S in the same way, and third, by the fact that the segregation of the disease in the families was not taken into account. However, we could not find any model that could explain the familial segregation of the trait, namely the higher proportion of affected parents than affected sibs. The aim of this work is to study and model the marginal effect of one susceptibility locus involved in the determinism of Kofendrerd Personality Disorder (KPD) in the Aipotu population. The presence of a susceptibility locus closely linked to the marker C03R0281 was shown by the existence of both strong association and genetic linkage between this marker and the trait [We want to select the replicates that best represent the distributions in the region of marker M (C03R0281) in terms of both association and linkage. Estimating the parameters and their variation intervals in this sample is then equivalent to evaluating them in the whole set of replicates. In the pooled sample set , we consider one index (an affected case) by family and his genotype for the marker M. For each index, we also consider his identity-by-descent (IBD) sharing for M with one random affected sib (families of the Aipotu population have been selected as having at least two sibs affected with KPD). In order to have a reliable IBD sharing for each sib pair, we ordered one SNP packet (153) surrounding marker M. The IBD sharing is obtained by maximum likelihood estimation using the information provided by the whole set of markers in the M region on chromosome 3.Figure Because we are looking for the model that best explains these four independent distributions (one genotype distribution and three stratified IBD distributions), we first determined the replicates that best reflect these distributions. We computed the distance of each of the 100 replicates to the pooled sample by a chi-square statistics, equal to the sum of the four independent chi-squares obtained by comparing the distributions observed in the replicate and in the pooled sample.We modeled the effect of the susceptibility factor by the MASC method. For a given genetic model, the MASC method computes the four expected distributions described in Figure We implemented a bootstrap procedure for calculating the variation intervals of the parameter estimates in the MASC program. The uncertainty on the parameters is due to the sampling of families and of controls, when the marker allele frequencies are inferred from a control sample. For each bootstrapped family set , we estimated the parameters considering the three possibilities described above for the marker allele frequencies. In situation 1, there is no uncertainty induced by the marker allele frequencies. In situation 2, the bootstrap procedure is applied to both the family sample and a sample of 50 controls randomly drawn among the 100 control samples. In situation 3, the bootstrap is only applied to the family sample. For the three situations, we obtain the distribution of the parameter estimates and provide the 95% intervals.Table 11and c12, whereWe tested a biallelic susceptibility locus model  and estimate four parameters: 2 relative penetrances, λ1 and λ2, and 2 coupling probabilities, cλ1 = P(affected | S1S2) / P(affected | S1S1),λ2 = P(affected | S2S2) / P(affected | S1S1),11 = P(S1 | M1),c12 = P(S1 | M2).cThe frequency (q) of allele S1 at the susceptibility locus can be written as11 P(M1) + c12 P(M2).q = P(S1) = c2 = 14.14; 6 df).The direct effect of marker M, which means S1 is confounded with M1, and S2 with M2, was rejected . The larger the sample size, the closer the estimate to the true value.Note that the true value of the dominance parameter cannot be inferred from the provided answers without extensive work. Indeed, the KPD phenotype is a mixture of different phenotypes, each one corresponding to different models of interaction between D2 and another susceptibility locus. Because there is no generation effect in the simulation, we still cannot explain the greater risk for parents than for sibs.GAW14: Genetic Analysis Workshop 14IBD: Identity by descentKPD: Kofendrerd Personality DisorderLD: Linkage disequilibriumMASC: Marker association segregation chi-squaresSNP: Single-nucleotide polymorphismMB performed the MASC and bootstrap analyses and drafted the manuscript. A-LL carried out the selection of the SNPs and draft the manuscript. EC carried out the study of the LD in the region, CB the association studies, M-CB the linkage studies. FC-D conceived, designed, coordinated the study and helped to draft the manuscript.


The critical role of inappropriate inflammation is becoming accepted in many diseases that affect man, including cardiovascular diseases, inflammatory and autoimmune disorders, neurodegenerative conditions, infection and cancer.This review proposes that cancer up-regulates the angiotensin II type 1 (AT1) receptor through systemic oxidative stress and hypoxia mechanisms, thereby triggering chronic inflammatory processes to remodel surrounding tissue and subdue the immune system. Based on current literature and clinical studies on angiotensin receptor inhibitors, the paper concludes that blockade of the AT1 receptor in synergy with cancer vaccines and anti-inflammatory agents should offer a therapy to regress most, if not all, solid tumours.With regard to cancer being a systemic disease, an examination of supporting evidence for a systemic role of AT1 in relationship to inflammation in disease and injury is presented as a logical progression. The evidence suggests that regulation of the mutually antagonistic angiotensin II receptors (AT1 and AT2) is an essential process in the management of inflammation and wound recovery, and that it is an imbalance in the expression of these receptors that leads to disease.In consideration of cancer induced immune suppression, it is further postulated that the inflammation associated with bacterial and viral infections, is also an evolved means of immune suppression by these pathogens and that the damage caused, although incidental, leads to the symptoms of disease and, in some cases, death.It is anticipated that manipulation of the angiotensin system with existing anti-hypertensive drugs could provide a new approach to the treatment of many of the diseases that afflict mankind. Tumour has been linked with inflammation since 1863, when Rudolf Virchow discovered leucocytes in neoplastic tissues and made the first connection between inflammation and cancer , includiSeveral reports implicate inflammation as a significant risk factor in cancer development: asbestos, cigarette smoke and inflammation of the bowel and pancreas are but a few well-known examples given ,10. ThesIt is evident that cancer is using natural inflammatory processes to spread and, unlikely as it seems at first, it is proposed that this is through the use of the angiotensin II type 1 (AT1) receptor.Angiotensin II (Ang II) is a peptide hormone within the renin-angiotensin system (RAS), generated from the precursor protein angiotensinogen, by the actions of renin, angiotensin converting enzyme, chymases and various carboxy- and amino-peptidases . Ang II AT1 receptors are expressed in various parts of the body and are associated with their respective functions, such as blood vessels, adrenal cortex, liver, kidney and brain, while AT2 receptors are highest in fetal mesenchymal tissue, adrenal medulla, uterus and ovarian follicles . The oppet al (2003) [In addition to the mediators reviewed by Suzuki l 2003) , a numbel (2003) , Tumour l (2003) , Plasminl (2003)  and adre03 , a nul (2003) ,22 and al (2003) . It has l (2003) .On examination of the tumour environment, it is interesting to note that angiotensin II actually increases vasodilation, a phenomenon that researchers have attempted to utilise for drug delivery . This woIn the 'Hallmarks of Cancer', the authoritative work by Douglas Hanahan and Robert A. Weinberg, the evolutionary acquired capabilities necessary for cancer cells to become life-threatening tumours are described. Furthermore, it is suggested that cancer researchers should look not just at the cancer cells, but also at the environment in which they interact, with cancers eliciting the aid of fibroblasts, endothelial cells and immune cells .Sustained angiogenesis, tissue invasion and metastasis are the latter of six necessary steps in tumour progression, as described in the 'Hallmarks of Cancer' . These eThe alternative basis under which induction of AT1 in tumours may occur is by looking at the environment under which the cancer is developing. Stresses and cell damage on the growing tumour boundary could potentially be causing the expression of AT1. Evidence that appears to support this view can be found in a study of AT1 expression in breast cancers . In thisThe formation of oxidised LDL by monocytes and macrophages at the sites of tissue damage has been established in a recent report by Jawahar L. Mehta and Dayuan Li . In thisHypoxia has been demonstrated to induce the expression of both AT1b (AT1a and AT1b are subsets of AT1) and AT2 receptors in the rat carotid body and pancreas ,40. The A review of hypoxia and oxidative stress in breast cancer cites the chaotic flow of blood in the tumor environment with resultant periods of hypoxia and reperfusion . ReperfuThe evidence relating to over-expression of AT1 with cancer progression is compelling. To this effect, AT1 blockade has been hypothesised as the mechanism to overcome cancer associated complications in organ graft recipients . AdditioTumour progression has been significantly slowed with AT1 receptor antagonists ,48. The The powerful direct and indirect suppression effects of TNF-α , IL-1β  and TGF-Despite this, it has long been established that the body does have the capability to recognise cancer cells and develop antigens. Dentritic cell vaccines for instance have been developed and have demonstrated limited effect in treating established tumours. The effectiveness of one such approach was greatly enhanced leading to complete regression of tumours in 40% of cases when TGF-β was neutralised using TGF-β monoclonal antibodies in synergy with a dentritic cell vaccine .Strong evidence suggests that tumour cells over-express AT1 receptors and compelling evidence has been presented on the implications of AT1 in cancer progression. Although still at a theoretical stage, this evidence leads to the formulation of the hypothesis that effective blockade of AT1 with a tight binding receptor antagonist, in combination with NSAIDs to further control the inflammation, and immunotherapy, such as cancer vaccines, would provide an effective treatment. Most, if not all, solid tumours utilise inflammation processes, which, through the over-expression and activation of AT1 and the subsequent expression of a number of inflammatory cytokines and chemokines, allow for tumour protection from the immune system through immunosuppression, as well as tumour progression and metastasis. Blocking these pathways through inhibition of AT1 using one of the commercially available AT1 inhibitors, whilst lifting the induced protective effect of immunosuppression and further reducing inflammation with the use of NSAIDs will both inhibit tumour progression and allow currently developed immunotherapies, such as cancer vaccines, to promote their therapeutic effect uninhibited. The role of AT1 post-metastasis, given the observation that AT1 protein expression ceases, as demonstrated in the breast cancer study, requires further investigation . HoweverCancer is a systemic disease, one that can affect every part and organ in the body and, as presented in this review so far with regards to the role of AT1 in cancer, AT1 upregulation is of the utmost importance in the activation of inflammation. Systemically, therefore, what purpose does this upregulation of AT1 serve? The release of ACE and extended expression of AT1 and AT2 during the healing process following vascular injury helps to answer this question . AngII iThe activation of AT1  and circulates in the blood (decreasing with age) and is important in the regulation of immunity and inflammation : IGF-1 iSignificant evidence has been shown that AT1 receptors are upregulated during disease and that AT2 receptor expression follows behind AT1 expression during injury and healing. Given the opposing roles of AT1 and AT2 it can thus be postulated that the interplay of these receptors plays a significant part in judging the current local status of appropriate versus inappropriate inflammation and in providing feedback to the rest of the body. Indeed it is anticipated that prolonged expression of AT1 combined with a lack of AT2 expression results in sustained chronic inflammation and fibrosis.Overall, the role of the AT receptors in managing and monitoring the healing process is complex, with many positive and negative feedback mechanisms both within the site of inflammation/healing and with the rest of the systems in the body. An attempt to summarise these systemic signalling inter-relationships is given in figure The invasiveness and immunosuppression of many cancers appears dependent on inflammation and the upregulation of AT1. Two mechanisms for upregulation of AT1 are discussed: 1) evolutionary changes to take advantage of this pro-inflammatory control mechanism, 2) AT1 expression induced by an alternating environment of hypoxia and oxidative stress. Immunosuppression as a common protection mechanism of solid tumours against immune responses has been verified from current literature and experimental procedures, as has the implication of cytokines and chemokines in tumour growth and metastasis. Given the involvement of AT1 in the immunosuppression and inflammatory processes, as well as in the expression of the pro-inflammatory cytokines and chemokines, it becomes evident that the AT1 receptor is essential for tumour protection and progression. A combination therapy consisting of AT1 receptor antagonists, NSAID for further control of the inflammation and immune therapy in the form of tumour vaccines should provide a novel and successful treatment for solid tumours.In the renin-angiotensin system, the angiotensin II receptors AT1 and AT2 seem to have opposing functions. The actions of AT1 being principally pro-inflammatory whilst AT2 provides protection against hypoxia, draws inflammatory action to a close and promotes healing. The various direct and indirect mechanisms for feedback between the receptors, their induced products and the external hormonal system in the control of inflammation and healing are summarised in a highly simplified model which none the less can be used to explain how many key promoters and inhibitors of disease exert their effects.From a review of the current disease literature, it has been demonstrated that the role of AT1 and AT2 in inflammation is not limited to cancer-associated inflammation, but is generally consistent and system wide. Potential therapy by manipulation of these receptors, although at an early stage, has been demonstrated for some of these diseases and it is proposed that this approach will provide an effective basis for the treatment of autoimmune, inflammatory and neurodegenerative disorders using existing drugs. AT1 receptor blockade should, in addition, provide a treatment to alleviate the damage caused by bacterial and viral infections, where their destructive action is through chronic inflammation. Given the importance of the immune suppressant effect of inflammation in cancer, it is anticipated that AT1 blockade should also serve to elicit a more effective immune response to other invaders that seek to corrupt the wound recovery process.Manipulation of the AT1 and AT2 receptors has profound and exciting implications in the control of disease.TGF-β Transforming Growth Factor BetaAT1 Angiotensin II Type 1 receptorAT2 Angiotensin II Type 2 receptorIGF-1 Insulin-like Growth Factor 1LOX-1 Lectin-like Oxidized Low-Density Lipoprotein Receptor 1HIF-1a Hypoxia Induced Factor 1 AlphaHMG CoA 3-Hydroxy-3-Methyl-Glutaryl Coenzyme AbFGF basic Fibroblast Growth Factor2-.)ROS Reactive Oxygen Species  in nature, our long-term ambition is to identify additional drug targets and agents that could work in combination with ACE inhibitors and AT1 blockers to treat most diseases.Sotiris Missailidis is a Lecturer at the Chemistry Department of The Open University, with research focus on cancer and had been the academic supervisor of Gary R Smith. There are no conflicting interests or financial implications related to the publication of this review article.Gary R Smith performed the literature review and proposed the hypothesis that cancer utilises the Angiotensin system to trigger chronic inflammation as a means of spreading and avoiding the immune system. In addition to providing significant editorial contributions and literature related comments, Sotiris Missailidis prompted Gary R Smith to undertake additional research with led to clarification of the role of hypoxia and oxidative stress in governing AT receptor expression. This understanding led Gary R Smith to propose the hypothesis that inflammation through the AT receptors is the cause of many of the diseases that affect mankind, including infectious diseases, which utilise inflammation to disrupt the immune system.


It remains unknown whether primary ocular adnexal extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is a homogeneous entity, as there are few reports of the results of cytogenetic or molecular analyses of these tumors.in situ hybridization (FISH) analysis to detect translocations and aneuploidy in 34 cases of primary ocular adnexal MALT lymphoma, and reviewed the histopathological findings. Correlations between the results of FISH analysis, the histopathological features and the clinical data were also analyzed.We performed interphase fluorescence Among the 34 cases, FISH analysis revealed t in one case, trisomy 3 in 21 cases (62%), and trisomy 18 in 16 cases (47%). The cases with trisomy 18 had significantly more prominent lymphoepithelial lesions (LELs) and less nodularity in the tumors. In regard to the clinical correlations, tumors with trisomy 18 were observed predominantly in females and younger patients; also, in the majority of the cases, the tumor was of conjunctival origin. All the cases with recurrence showed trisomy 18 in the tumor.Primary ocular adnexal MALT lymphoma is a significantly heterogeneous entity. Cases with trisomy 18 may have unique clinicopathological features. Since the first description by Isaacson and Wright in 1983, MALT lymphoma has been recognized as a distinct entity of low-grade B-cell lymphoma; it is described as a separate entity in the revised European-American lymphoma (REAL) classification and also in the more recent classification by the World Health Organization (WHO) [Various cytogenetic changes, such as t11;18)q21;q21), and t1;14)p22;q32) and trisomy 3, 7, 12 and 18, have been detected as recurrent chromosomal aberrations in this tumor [1;q21, an2 and triMalignant lymphomas arising from the ocular adnexa account for about 8% of all extranodal lymphomas . They haArchived samples of primary ocular adnexal MALT lymphoma at our institute between 1995 and 2003 were utilized for this study. The diagnosis of MALT lymphoma was confirmed by histopathological examination of either biopsy specimens or surgically resected specimens of the primary ocular adnexal lesions. In all the cases, the diagnosis was reviewed by two hematopathologists (YM and AAM) according to the REAL classification or the WHO classification. Samples for the FISH analysis were obtained from the same primary lesions in the ocular adnexa.This study was approved by the institutional review board of National Cancer Center (16–7).The clinical data were obtained from the medical records. The sex, age, clinical stage, site of involvement, performance status (PS) according to the Eastern Cooperative Oncology Group scale, the serum lactate dehydrogenase (LDH) value at the time of initial presentation, and the initial therapy were recorded. The anatomic site of involvement was determined by the system reported by Knowles et al. . We specThe tissue FISH procedure was performed in formalin-fixed paraffin sections in accordance with the procedure described by Sekiguchi et al. . We applWe also analyzed 4 reactive lymphomatous lesions of the ocular adnexa as a negative control and the threshold was determined for each probe by counting the number of signals or fusions in one nucleus per 100–200 cells. The signal frequency thresholds were determined as the mean plus 3 standard deviations. Each specimen was examined independently by three investigators . The number of signals or fusions in one nucleus was counted in 200 cells, and when the results were judged to be the same, the specimen was considered as being successfully stained and included in this study.Each hematoxylin eosin (HE)-stained specimen was histologically reviewed by two of us (KT and YM). In addition to histopathology, the paraffin-embedded sections were also subjected to immunostaining for CD3 by the standard avidin – biotin method using monoclonal anti-CD3 antibody  as the primary antibody. Immunophenotypic analysis revealed that the tumor cells of MALT lymphoma were positive for CD20 and negative for CD5, CD10 and cyclin D1. Frequently but not universally observed morphological features of primary ocular adnexal MALT lymphomas were examined for and categorized as follows: the finding of an abundance of plasma cells, monocytoid B cells and reactive CD3-positive T cells in none of the areas of the tumor was denoted as negative (-), in less than 30% of the tumor area was denoted as focally positive (+), in 30–60% of the tumor area was denoted as positive (2+), and in 60% or more of the tumor area was denoted as strongly positive (3+). The finding of residual reactive germinal centers, large lymphoid cells, polykaryocytes, brown-pigment-laden histiocytes, mitotic cells, lymphoepithelial lesions (LELs) and Dutcher bodies was categorized as negative (-), focally positive (+), or positive (2+) when it was absent, present but inconspicuous, or very conspicuous, respectively. The degree of nodularity was classified as negative (-) in cases without discernible vague nodularity, focally positive (+) in cases with vague nodularity, and positive (2+) in cases showing definite nodularity.Statistical analysis was performed using the SPSS software for Windows, version 13.0. The correlations between the clinical features, results of FISH analysis and the morphological features were analyzed by Fisher's exact test. Survival was censored at the time of the last documented follow-up date. The overall survival period and time to recurrence were calculated using the Kaplan-Meier method and compared by the log-rank test.The characteristics of the patients are shown in Table The most frequently involved site was the orbit (62%). In 26 patients (76%), the disease was unilateral, whereas in eight patients (24%) it was bilateral. Almost all patients had localized disease and a serum LDH in the normal range. None of the patients had any manifestations of autoimmune diseases. The initial therapies varied among the patients: 21 patients (66%) received radiation only, three (9%) received chemotherapy, three (9%) received chemotherapy combined with radiation therapy and 5 (16%) were observed under a watch-and-wait policy. Two patients were lost to follow-up after the diagnosis. After a median follow-up duration of 31 months , all 32 patients were alive and the disease recurred in five patients. There were no significant differences in terms of the sex or age of the patients, site of involvement, laterality, initial therapy, stage, or the serum LDH level at the time of initial presentation (data not shown).The signal frequency thresholds were determined as the mean plus 3 standard deviations (SD). The mean values of more than two signals for the CEP 3, 7 and 18 probes were 2.50%, 0.67% and 1.75%. The thresholds of the CEP 3, 7 and 18 probe signals were 4.23%, 2.40%, and 3.25%. IgH/MALT1 and API2/MALT1 fusions and more than two signals for CEP 12 probe were not observed for any of the 4 control samples. Based on these results, we decided to set a cut-off value of 5% for all the probes to reduce the incidence of false-positive results.The results of FISH analysis are shown in Table The relationship between the clinical factors of the patients and the detection of trisomy 18 is shown in Table The disease recurred in five cases, and all of these cases had trisomy 18. As shown in Fig. Table We attempted to identify the existence of any correlation between the cytogenetic heterogeneity as revealed by FISH analyses and the morphological features. Table A number of karyotypic abnormalities have been reported in MALT lymphomas. Among these, t was the most frequently encountered structural chromosomal abnormality resulting from the fusion of the apoptosis inhibitor-2 (API2) gene and the MALT1 gene at the 11q21 and 18q21 breakpoints ,20-22. SThe results of the FISH analysis revealed that translocations such as t11;18) and t were rather rare; only one case showed t. In the study by Streubel et al., nine out of the 37 cases 24%) of ocular adnexal MALT lymphoma were positive for t and one out of the 37 (3%) cases were positive for t according to the results of FISH analysis [% of oculIn contrast, we found that aneuploidy was rather common. Approximately half of all the tumors showed trisomy 3 and/or 18. The frequency was at the upper end of the previously reported range of 10–50% ,8,24. ThThe ocular adnexal MALT lymphomas also exhibited morphologic heterogeneity, and the degree varied. Several reports revealed that the majority of ocular adnexal lymphomas were the MALT type. To the best of our knowledge, there have been no detailed analyses of the histopathological characteristics of ocular adnexal MALT lymphomas -16. We sThe differences in the morphological features of the tumors were related to the primary site of involvement of the MALT lymphomas; the number of large lymphoid cells, polykaryocytes and LELs, and the degree of nodularity of the tumor varied significantly depending on the site of origin of the tumors. It may only seem natural that tumors arising from different sites show different morphological characteristics; however, there have been no reports focusing on this issue among ocular adnexal MALT lymphomas, partly because of the small number of patients. The morphological differences suggest that MALT lymphomas of ocular adnexa origin might represent a combination of several diseases which may need to be analyzed separately.The morphological features were also related to the chromosomal aberrations; more specifically speaking, the presence of trisomy 18 was associated with a specific morphology. The tumors in cases with trisomy 18 had more LELs and less nodularity than those without trisomy 18. In addition, we found that the cases with trisomy 18 had shorter recurrence-free survivals than those without trisomy 18. These five cases showed no significantly different characteristics from the other cases in terms of the sex or age of the patient, the site of involvement, laterality, initial therapy, stage, or the serum LDH levels at the time of initial presentation. Trisomy 18 was strongly related to the recurrence-free survival rate. These results show that tumors with trisomy 18 may represent a characteristic and one unique subgroup among primary ocular adnexal MALT lymphomas. This recognition might compare well with the situation of gastric MALT lymphomas with t, which constitutes a specific subgroup in terms of the clinical setting, patient prognosis, and morphological findings among all cases of gastric MALT lymphoma ,26. The Recently, it has been reported that some ocular lymphomas may be related to Chlamydia and hepatitis C virus infection, and the involvement of these agents might be the reason for the heterogeneity among the tumors -29. In oWe detected aneuploidy in cases of ocular adnexal MALT lymphomas, and also morphological heterogeneity. These results demonstrate that ocular adnexal MALT lymphomas differ in characteristics from those at other anatomic sites, and also show significant heterogeneity. Also, we suggest that these tumors showing trisomy 18 might represent a unique entity.The author(s) declare that they have no competing interests.KT, NS, YY and YK carried out the FISH analysis. KT, AMM and YM carried out the histological analysis. KT, NS, AK, TW and KT participated in the design of the study and performed the statistical analysis. KT, NS, MH and YK conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:


The maintenance of standards is a problem for postgraduate medical examinations, particularly if they use norm-referencing as the sole method of standard setting. In each of its diets, the MRCP(UK) Part 1 Examination includes a number of marker questions, which are unchanged from their use in a previous diet. This paper describes two complementary studies of marker questions for 52 diets of the MRCP(UK) Part 1 Examination over the years 1985 to 2001 to assess whether standards have changed.Study 1, which used routinely collected information on the performance of 4405 marker items, used a statistical method to assess changes in performance across diets. Study 2 compared performances of individual candidates on 28 individual marker items that were shared by the 1996/2 and 2001/3 diets.Study 1 found evidence that candidate performance on the MRCP(UK) Part 1 Examination showed a gradual improvement over the period 1985 to 1997, which was followed by a sharp decline in performance until 2001. The 'dog-leg' in performance at 1997/3 was not an artefact of changed Examination Regulations, mix of UK and overseas candidates, or time from qualification until taking the Examination. Study 2 confirmed that performance in 2001/3 was significantly worse than in 1996/3, that the poorer performance was found in graduates of UK medical schools, and that candidates passing the Examination in 2001/3 performed less well than those passing in 1996/2.There has been a decline in the performance of graduates from UK medical schools taking the MRCP(UK) Part 1 examination. The reasons for this are not clear, but the finding has implications for medical education, and further studies are needed of performance in other postgraduate and undergraduate examinations. The use of norm-referencing as the sole method for setting the pass mark over this period meant that candidates passing the MRCP(UK) examination also had a lower standard. The MRCP(UK) Part 1 and Part 2 examinations now have their standard set by criterion-referencing. The unstandardised regression coefficients of vd  are then the estimates of sd , with sr = 0, and the standard errors of the regression coefficients are the standard errors of the estimates of sd .SPSS version 11.5 was used for statistical analysis of the data. For statistical analysis, diets 1, 2 and 3 in a year were set at 0, .33 and 0.67 of the year.Raw data were available only for the 1996/2 diet and the diets from 1997/1 onwards, and of these only the 1996/2 and 2001/3 diets shared sufficient marker items for analysis  Part 1 Examining Board as the performance of the top 35% of candidates taking the Examination in the UK on their 1All analyses were restricted to 'UK graduates', defined as those with primary registrable qualifications from United Kingdom medical schools.Altogether 5332 marker questions were used in the 43 diets held between 1988/1 and 2002/1, and these had previously been used on diets extending back to 1972. Although in principle the statistical method can assess standards outside of the range of the diets actually assessed, a preliminary analysis suggested the process was unstable prior to about 1985, with large standard errors of the estimates. Analysis is therefore restricted to the 4405 marker questions that provided information on the performance of the 52 diets held between 1985/1 and 2002/1.Figure sd, calculated by regressing the dummy variables v1 to vn on the values of Δm. The ability of candidates increased slowly but consistently between 1985 and about 1996, after which ability appears to decline fairly steeply. That hypothesis was formalised by using non-linear regression to fit two straight lines to the data, each with its own independent slope, with a 'dog-leg' mid-way through the data, the date of the dog-leg itself being a free parameter. The dog-leg curve shown in figure 2 = .468), and is a significant improvement over a simple linear regression  = 18.28, p < 0.001). The inflexion of the dog-leg is at 1997.3 , with 95% confidence intervals of 1996.0 to 1998.6 . The slope before 1997 is +0.18%/year (95% CI 0.03% to 0.33%), and the slope after 1997 is -1.34%/year (95% CI -1.93% to -0.75%).The individual points in figure The 1996/2 diet was taken by 2132 candidates of whom 852 were UK graduates. The 2001/3 diet was taken by 2051 candidates of whom 557 were UK graduates. The 28 marker items were combined into a single scale for which alpha was 0.593, which is equivalent, using the Spearman-Brown formula , to a reIn the 1996/2 diet, the 852 UK graduates have a mean marker score of 69.4 (SD 14.5), compared with a mean marker score of 59.6 (SD 15.8) for the 557 UK graduates in the 2001/3 diet . The average mark in 2001/3 was therefore 14.1% lower than in 1996/2 when precisely the same items are compared. Detailed analyses of items answered correctly, incorrectly or not answered showed that the 2001/3 candidates had answered fewer of the 28 items correctly  and more of the items incorrectly , and more were not answered . The reduced performance in the 2001/3 diet was independent of whether candidates were taking the Examination on their first, second or subsequent attempts  on 17 of the 28 individual marker items  Part 1 examination for the twelve years between 1985 and 1997. Such a change could either result from a genuine increase in candidates' knowledge or from candidates becoming more 'test-wise', perhaps due to attending crammer courses and becoming more aware of questions from previous papers. However, the latter explanation seems unlikely in view of the sharp decline in performance of candidates after 1997.Additional File, have investigated the following possibilities:If the decline in performance after 1997 reflects decreasing candidate ability, then that has important implications for medical education and training, and possible confounders and artefacts must be excluded. Separate analyses, not reported in full here but available in the From 1999/2 onwards, candidates were not restricted to four attempts at the Examination. Figure The MRCP(UK) Part 1 examination was negatively marked, wrong answers incurring a greater penalty than unanswered questions. If candidates changed their strategy, that may have produced an apparent decline in performance. We have modelled the number of not-answered questions, and although this did show a decline in between 1990 and 1993, the timing was unrelated to the change shown in Figure Additional File).Because the marker question statistics recorded in the Examination records are based on aggregate statistics for all candidates taking the examination, changes in the mix of candidates could cause a change in marker question performance. We have looked at the relative proportion of UK and overseas graduates, the numbers of candidates on first, second, third, fourth and later attempts, and the time between qualification and taking the MRCP(UK) Part 1 Examination, and although secular trends are visible, none shows a sudden change in 1997.  Examination may have changed over time, rising gradually until about 1997, and then declining rather more rapidly. The discontinuity around 1997 does not seem to be related to any obvious change in the structure of the Examination or the composition of the candidates taking it.Study 2 examined the performances of individual candidates on marker items and confirms the earlier finding that performance dropped between 1996/2 and 2001/3. In particular, fewer UK graduates who passed the Examination in 2001/3 gave correct answers than had equivalent candidates in 1996/2. The median change in performance on items of -7.75% was similar to the expected change of -7.1%, based on the -1.34% per year shown in figure We found that the performances on individual marker items between 1996/2 and 2001/3 had dropped for 21 of the 28 items. None of the questions is about recondite, obscure or unimportant areas of knowledge for a general physician in training, and none of the changes are likely to reflect changes in the importance of knowledge, in understanding of disease mechanisms, or in treatment strategies. They are therefore acceptable marker questions. The largest decreases were on the electrocardiography and anatomy of the Wolff-Parkinson-White syndrome, aseptic meningitis in infectious mononucleosis, and bone marrow biopsy findings in the anaemia of chronic infection, all of which are important clinical problems. The only two significant increases in knowledge are on questions concerning C-reactive protein , and on the diagnosis of organic brain disease.These two complementary studies have implications specifically for postgraduate medical examinations, and more generally for undergraduate medical education. The studies provide evidence that there was a sudden, relatively steep decline in the performance of candidates passing the MRCP(UK) Part 1 Examination between 1997 and 2002, which was not an artefact of changes in the mix of overseas and UK candidates, or changes in the time after qualifying of first or subsequent sittings of the examination. Study 2 confirmed that the decline had taken place in doctors graduating from UK medical schools.The MRCP(UK) examination sets a standard for professional clinical practice in the UK. Our use of marker questions for assessing the standard across diets parallels a study in 1989, which described the falling standard of candidates passing the American Board of Internal Medicine (ABIM) examination   Part1 and Part 2 written Examinations have recently changed their format, the Part 1 Examination now consisting entirely of 'best-of-five' questions. The MRCP(UK) Part 1 and Part 2 Examinations both carry out standard-setting by a process incorporating criterion referencing using the Angoff technique ,13, withThe decline in performance of candidates from UK medical schools taking the MRCP(UK) Part 1 Examination raises questions extending beyond the Part 1 Examination itself. The examination can be taken eighteen months after graduation, and a high proportion of UK graduates take it at the earliest possible time, when they typically have five or six years of undergraduate education, a year of PRHO posts, and six months of SHO training. Several explanations need to be considered for the changes in standard that we have found.Topics once perceived as central to medical training may now no longer be important to modern medical practice. If the marker questions used were out-of-date then that may explain the apparent decline. However, not only is the content of marker questions always approved by MRCP(UK) Part 1 Examining Board before each inclusion in the Examination, but the questions shown in table The present results relate only to one examination, the MRCP(UK) Part 1 Examination, albeit an exam taken by over 30% of UK graduates. Corroboration of the present findings from other UK postgraduate examinations is desirable, in order to assess the generality of the findings. It is possible that around 1997 more able UK graduates candidates decided they no longer wished to take the MRCP(UK) Part 1 Examination, and instead took other career paths (and that seemed to be the explanation for the declining standard in the ABIM examination ). AlthouIn recent years the working hours of junior doctors have declined, in part due to changes in Government regulations, and clinical experience and hence examination performance may also have declined. In the absence of good measures of clinical experience this hypothesis is difficult to test. There is, however, evidence that the undergraduate clinical experience of UK doctors qualifying in 1996 was lower than that of doctors qualifying a decade earlier , and thaTomorrow's Doctors [Tomorrow's Doctors. A key question concerns whether the standards of undergraduate examinations, both basic medical sciences and finals, have been maintained; however, the patchy use of marker questions, frequent changes in undergraduate examination formats, and the absence of a UK national medical licensing examination make it unlikely that the question can be answered easily. Indeed, the only reliable evidence on the absolute standard of undergraduate training may have to come from performance in postgraduate examinations.Undergraduate medical training in the UK has been continually changing for nearly four decades, dating back primarily to the Royal Commission of 1968 , and supIn summary, we have provided evidence of a decline in the performance of candidates taking the MRCP(UK) Part 1 Examination between 1997 and 2001. In addition, as a result of the reliance on norm-referencing, there was also a decline in the standard of those passing the Examination. Criterion referencing is now included as a central part of the MRCP(UK) Examination standard setting process. The reasons for the declining standard of UK graduates are not clear, but on balance are more likely to reflect changes in undergraduate training than changes in postgraduate medical education. More research into the standard of other postgraduate examinations, as well as undergraduate assessments, is urgently needed.ICM, JAV, OLD and JM have all been involved with the MRCP(UK) for many years. JM is a paid employee of the MRCP(UK) central office, JAV, OLD, and ICM contribute to the examination academically, and JAV and OLD receive financial compensation for the extensive work that this involves.The original idea for the study was ICM's, arising from discussions that occurred at Board Meetings chaired by JAV. JM contributed extensive statistical and administrative support to the study, and JAV and OLD contributed to the interpretation of results and the writing of the paper. ICM wrote the first draft of the manuscript and carried out the multivariate statistical analyses. All authors have contributed to the final draft of the paper.The pre-publication history for this paper can be accessed here:Changes in the composition and timing of entries to MRCP(UK) Part 1.Click here for file


The body mass index (BMI) of preschool children from 4 years of age through primary school has increased since the Great East Japan Earthquake, but that of children aged under 3 years has not been studied. This study evaluated how the anthropometrics of younger children changed following the earthquake.Height and weight data of children living in northeast Japan were collected from 3-, 6-, 18-, and 42-month child health examinations. We compared the changes in BMI, weight, and height among infants affected by the earthquake between their 3- and 6-month health examinations, toddlers affected at 21–30 months of age (affected groups), and children who experienced the earthquake after their 42-month child health examination (unaffected group). A multilevel model was used to calculate the BMI at corresponding ages and to adjust for the actual age at the 3-month health examination, health examination interval, and gestational age.We recruited 8,479 boys and 8,218 girls living in Fukushima, Miyagi, and Iwate Prefectures. In the infants affected between their 3- and 6-month health examinations in Fukushima, the change in BMI at 42 months of age was greater than among the unaffected children. In the toddlers affected at 21–30 months of age in Fukushima, the change in BMI was greater, but changes in weight and height were less.Affected infants and toddlers in Fukushima suggested some growth disturbances and early adiposity rebound, which can cause obesity. The future growth of children affected by disasters should be followed carefully. Studies conducted after the Great East Japan Earthquake show that dietary sources were restricted during the acute phase,13 and outdoor physical activity decreased during the post-quake phase because of fear of exposure to radioactive discharge from damaged nuclear power plants.14 Fear of death may change hormone levels among those affected.15 Experts suggest that growth retardation during the first 2–3 years of life can cause undesirable health outcomes later in childhood.16,17 Studies on the impact of the earthquake have been limited to preschool children about 4 years of age,8 and little is known about the influence on children younger than 3 years of age. The aim of this study was to determine: 1) whether the growth of infants and toddlers was disturbed by the earthquake; and 2) how the earthquake changed the BMI of children living on the Pacific coast of northeast Japan.Following an earthquake, micronutrient deficiency in children during the acute phase18 In total, 31 out of 59 municipalities (52.5%) in Fukushima Prefecture, 19 out of 36 (54.3%) in Miyagi Prefecture, and 30 out of 33 (90.9%) in Iwate Prefecture collaborated with this survey and provided data from 3-, 6-, 18-, and 42-month child health exams. During the healthcare visits, the children’s gestational age and birth weight were collected through an interview or from a maternal and child health handbook, and their weight and height were measured. We compared the BMIs of three study groups: the ‘group affected between 3 and 6 months old’ included infants who experienced the earthquake between their 3- and 6-month health exams; the ‘group affected between 21 and 30 months old’ included children between 21–30 months of age at the earthquake and between their 18- and 42-month health exams; and the ‘unaffected group’ (reference group for comparison) included any child who had completed a 42-month health exam before the earthquake. This last group’s anthropometric data were not considered to have been affected by the earthquake. Through the municipalities, we invited 20,600 children from Fukushima, 15,804 from Miyagi, and 17,932 from Iwate who were born from March 2007 through April 2011. At the baseline of the 3-month health exam, 8,855 boys and 8,508 girls in Fukushima, 4,293 boys and 4,041 girls in Miyagi, and 6,851 boys and 6,714 girls in Iwate were included. The data with analyzable values included 2,890 children from Fukushima, 461 from Miyagi, and 2,309 from Iwate in the group affected between 3 and 6 months old; 2,331, 422, and 1,508 in the group affected between 21 and 30 months old; and 3,212, 1,190, and 2,374 in the unaffected group, respectively. These figures constitute around 13.9% of children in Fukushima, 2.6% in Miyagi, and 15.2% in Iwate, according to the numbers of children enrolling in primary school in the following years.19With municipal government assistance, we invited children living along the Pacific coast of northeast Japan  to participate in a survey on infant growth. The details of the Health Examination Survey on Early Childhood Physical Growth in the Great East Japan Earthquake Affected Areas have been published elsewhere.The primary outcome in the group affected between 3 and 6 months old was the change in BMI (weight in kilograms divided by the square of height in meters) from a baseline age of 3 months to 42 months old compared with the unaffected group. The primary outcome in the group affected between 21 and 30 months old was the change in BMI from a baseline age of 18 months to 42 months old compared with the unaffected group. The secondary outcomes in the two affected groups were the changes in weight and height compared with the unaffected group. The data of the children who received each of the 3-, 6-, 18-, and 42-month health exams were analyzed for the group affected between 3 and 6 months old. The data of those who received both the 18- and 42-month health exams were analyzed for the group affected between 21 and 30 months old.20 was used to adjust for the level of statistical significance, correcting for comparisons at three time points between the groups affected between 3 and 6 months old and not affected. The P-values were evaluated at one-third of the level of the usual statistical significance. For instance, we applied a singular multilevel model to estimate the changes in the BMI from the age of 3 to 6, 18, and 42 months in the children affected between their 3- and 6-month exams and the unaffected children by strata of prefecture and sex. In the same model equation, we estimated the change in BMI from the age of 18 to 42 months in the children affected at between 21 and 30 months old. The estimated coefficients were used to calculate the adjusted changes in BMI at an actual age of 6, 18, and 42 months. A total of 12 models were used for the boys and girls of Fukushima, Miyagi, and Iwate Prefectures. The model for the change in BMI was:ij is the residual term in the model. We plotted the mean adjusted change in BMI at 6, 18, and 42 months of age as calculated using the model coefficients.To determine the influence of the earthquake on continuously growing children, we estimated and compared the changes in the anthropometrics from 3 to 6, 18, and 42 months of age between the children affected between their 3- and 6-month health exams and the unaffected children. We also compared the changes from 18 to 42 months of age between the children affected at between 21 and 30 months old and the unaffected children. We used a multilevel model  to adjust for the gestational age, actual age at the 3-month health exam, and the intervals between the 3-month and subsequent exams. Bonferroni’s methodP-value of the coefficients (β1). All statistical analyses used SAS statistical software . Descriptive statistics were reported as the means and standard deviations (SD). All reported P-values were two-sided; P-values <0.05 were considered to be statistically significant.For sensitivity analyses, changes in BMI were estimated using a similar model, with adjustment of the birth weight or BMI at the 3-month health exam, instead of the gestational age. In the plotted figure, the statistical significance of the differences between each affected group and the unaffected group was assessed using the The ethics committee of the Tohoku University School of Medicine approved the study protocol (no. 2012-1-125). The study was conducted following the ethical guidelines and regulations of the Declaration of Helsinki. The data were collected anonymously, and the participants could choose to opt out of the study. This study was an analysis of the data of the municipal healthcare service, and informed consent was therefore not required for this investigation according to the Japanese guidelines.21 Table Table P = 0.0006) and 42 months (P < 0.0001) of age and for the girls at 42 months of age (P = 0.0005) compared with the unaffected group. In the group affected between 21 and 30 months old in Fukushima, there were significant increases in the BMI from baseline for both boys (P = 0.014) and girls (P = 0.0034) at 42 months old, compared with the unaffected group. In the group affected between 3 and 6 months old in Miyagi, there were significant decreases in the changes in BMI for the boys at 6 (P < 0.0001) and 18 months old (P = 0.0005) compared with the unaffected group. In the group affected between 21 and 30 months old in Miyagi, there was no significant change in the BMI of either boys or girls compared with the unaffected group. In the groups affected between 3 and 6 months old and 21 and 30 months old in Iwate, there were no significant changes in the BMI of either boys or girls compared with the unaffected group. These results were consistent with analyses of the changes in BMI in similar models with adjustment for birth weight and BMI at the 3-month health exam  were relatively small. Since a subset of residents left the prefectures, the accurate proportion of children in each prefecture is unknown. However, although the anthropometric data did not include severely-affected infants who moved away or died, a recent analysis suggests that the influence of those who moved out and died might be ignored.38 In normally-growing infants, the growth in height catches up with the increase in weight. Particularly in Fukushima, the infants in the group affected between 3 and 6 months old exhibited unbalanced changes in mean weight and height (Table 39,40 We recommend that health administrators and pediatricians should carefully follow the affected children in Fukushima to assess their growth, as well as to check for the development of potential cardiovascular risks from adolescence to adulthood.In infancy, a decrease in BMI indicates a more rapid increase in height than weight.The infants affected at between 3 and 6 months old in Fukushima Prefecture showed an increase in mean BMI towards 42 months compared with the reference group, which suggests a risk of potential early adiposity rebound at a population level. The data also suggested the potential risk of this in toddlers affected between 21 and 30 months old in Fukushima, with a smaller increase in mean height and weight. In Miyagi Prefecture, the mean BMI of the male infants decreased immediately after the earthquake but recovered. The results underscore the role of pediatricians in following the growth of affected infants when disasters occur, to prevent obesity and its complications.


Not only are A2ARs involved in the mechanism of transactivation of BDNF receptor TrkB, they also modulate the effect of BDNF on synaptic transmission, playing a facilitatory and permissive role. The cAMP-PKA pathway, the main transduction system operated by A2ARs, is involved in such effects. Furthermore, a basal tonus of A2ARs is required to allow the regulation of BDNF physiological levels in the brain, as demonstrated by the reduced protein levels measured in A2ARs KO mice. The crucial role of adenosine A2ARs in the maintenance of synaptic functions and BDNF levels will be reviewed here and discussed in the light of possible implications for Huntington's disease therapy, in which a joint impairment of BDNF and A2ARs seems to play a pathogenetic role.In the last few years, accumulating evidence has shown the existence of an important cross-talk between adenosine A


This paper examines the effect of ultrasonic vibration (USV) on grain size and interrupted porosity in Gas Tungsten Arc (GTA) spot-welded copper. Grain size was refined by perpendicularly attaching a transducer to the welded sheet and applying USV to the weld pool for a short time  in addition improvements to the degassing process. Results illustrate a significant reduction of grain size (57%). Notably, USV provided interaction between reformations (fragmentation) and provided nucleation points (detaching particles from the fusion line) for grains in the nugget zone and the elimination of porosity in the nugget zone. The GTA spot welding process, in conjunction with USV, demonstrated an improvement in the corrosion potential for a copper spot-welded joint in comparison to the joint welded without assistance of USV. Finally, welding of copper by GTA spot welding in conjunction with ultrasound for 2 s presented significant mechanical properties. The market requirements of copper grow annually , especiaHowever, the presence of porosity at the copper NZ has been noticed in many fusion welding studies, and the grain size for weldments of pure copper metal are more significant than weldments of copper alloys. In this study, welding pure copper sheets by the GTA spot welding process is proposed and tested via experiment in which a transducer was attached underneath the lower plate. USV was initiated and interacted directly after the end of the main arc application. Various vibration times were assessed . Additionally, tensile shear tests, microstructure, electrical conductivity, and corrosion resistance were investigated. Significant attention was paid to the mechanism of enhancement and its effect on the investigated properties.3, 5 mL HCl, and a few drops of HNO3 and 100 mL water for a few seconds at room temperature. The temperature history of the HAZ was recorded at 12.5 mm from the center of NZ (3 mm from the ceramic cup) by a four-input data logger-type DT-3891G digital thermometer . Samples with dimensions of 30 mm × 25 mm × 2.4 mm were cut from a lapped joint then ground and polished for electrical conductivity testing with a D60K-E eddy current electrical conductivity measurement system . Corrosion potential and current at the cross-section of the welded joint were recorded on a Zahner IM6e electrochemical workstation  using a solution of 3.5 wt % NaCl. The standard calomel electrode and Pt were used as a reference electrode and auxiliary electrode, respectively. The polarization curve was recorded by means of potential-dynamic scanning with a scan rate of 1 mV/s and analyzed by Tafel straight-line extrapolation.Pure copper, 99.97%, with dimensions 100 mm× 25.4 mm× 1.2 mm was welded with GTA spot welding to achieve a fluxless and fillerless joint a. A tranIn this study, an ultrasonic transducer with a frequency 28 KHz and 60 W power was attached to copper welded plates proposed to refine and degas grains in the nugget zone.At temperatures above approximately 450 °C, absorbed hydrogen in molten copper reacts with the inclusions to form steam in the microstructure :Cu2O + The macroporosities can be interrupted at the fusion line beside the NZ, which represent a critical reason to initiate a fracture. Half of the GTA spot-welded copper joints were simulated with Abaqus CAE software which is symmetrical at two sides of the nugget. The simulation consisted of the application of an axial tensile load at the edge of the sample and an encastre boundary condition at the NZ contact surface. The fracture of the simulated spot-welded joint with nugget sizes of 5, 7.5, 10, 12.5 mm are shown in Macroporosities were increased on the surface with the presence of USV, as shown in Grain size in the nugget zone of welds was measured following the procedures as described in ASTM E112-13—Standard Test Methods for determining average grain size. The investigation of the cross-section of welded joints also showed the reduction of grain size in the HAZ and NZ . For welThe detached particles and fragmented grains were investigated. The distribution of porosity through the nugget zone was investigated. 2) was assumed to estimate the tensile-shear strength , C0 is 456 ms−1 ); and S Pa = 4.38 MPa; thus, bubbles rose to open at the surface of the molten metal.The estimated pore pressure was The primary application of pure copper is transference of electrical power. In this study, the electrical conductivity of the copper spot-welded joint was investigated. corr) decreased in the sequence of BM, 0 s, 4 s, 6 s, and 2 s, while the corrosion potential (Ecorr) increased in the sequence of BM, 0 s, 6 s, 2 s, and 4 s. Because the smaller Icorr denoted a slower corrosion rate and the higher Ecorr denoted greater difficulty to start the etching, the fresh grains at NZ and HAZ were subjected to ultrasonic peening. Accordingly, the Ecorr of the weld joints were higher than the BM and the welded joint without ultrasonic peening [In  peening . Local cUSV had no obvious effect on grain size at NZ. The vibration of NZ for 6 s reduced the grain size ~57%.The cooling rate of the welded metal increased for joints welded with the assistance of USV, which led to an increase of the heat dissipation rate from NZ to the surrounding space. Accordingly, it suppressed the grain growth at HAZ.The refining of grains at HAZ and NZ enhanced the tensile-shear strength of the copper arc spot-welded joints.The mechanism of grain refinement in GTA spot welding of copper assisted with USV was analyzed. It was concluded that USV dissolved the fusion line into small particles that detached to form the nucleation of the new grains at NZ.The mechanism of degassing of NZ was also analyzed. USV activated porosities to be joined and increased internal size and pressure, supporting the movement to be opened at the surface when the pressure of the bubble was 5 MPa.The peening action, degassing, and refining of grains reflected positively on the improvement of electrical conductivity and corrosion behavior of the GTA spot-welded copper joint.In the present work, the assistance of ultrasonic vibration time  for GTA spot-welded copper joints was investigated. The process showed a refining of the grain size at NZ and HAZ simultaneously. Conclusions from the results are as follows:


Moreover, we report evidence of a sudden and anomalous increase in sedimentation rate around 2600 yr BP, leading to the deposition of a 4-6m thick package of alluvial deposits in approximately one century. We discuss this datum in the light of possible tectonic activity along a morpho-structural lineament, revealed by the digital elevation model of this area, crossing the Forum Boarium and aligned with the Tiber Island. We formulate the hypothesis that fault displacement along this structural lineament may be responsible for the sudden collapse of the investigated area, which provided new space for the observed unusually large accumulation of sediments. We also posit that, as a consequence of the diversion of the Tiber course and the loss in capacity of transport by the river, this faulting activity triggered the origin of the Tiber Island.The Tiber valley is a prominent feature in the landscape of ancient Rome and an important element for understanding its urban development. However, little is known about the city’s original setting. Our research provides new data on the Holocene sedimentary history and human-environment interactions in the Forum Boarium, the location of the earliest harbor of the city. Since the Last Glacial Maximum, when the fluvial valley was incised to a depth of tens of meters below the present sea level,  To reconstruct details of the pre-urban landscape, a combination of deep-trench excavation, coring survey, environmental sampling and paleomagnetic analysis has allowed us to explore the archaeological and alluvial sequence of the Tiber River Valley to a depth of 55 m below the modern surface. Here, we present the results of our investigation of the river sedimentary history in this sector since the end of the Last Glacial Maximum, when the fluvial valley was incised to a depth of tens of meters below the present sea level, until the establishment of a relatively stable alluvial plain at 6–8 m a.s.l. during the late 3rd century BCE. This new reconstruction provides a starkly different context for the crucial period during which Rome was transformed from a hut settlement to a monumental city.Despite Rome’s prominent position in the development of Mediterranean civilization, scholars actually know frustratingly little about the city’s original setting and early history. The present cross-disciplinary investigation of Rome’s river valley has produced new data on the Holocene evolution of the landscape in the heart of the ancient city. This area, known as the Forum Boarium, is the location of Rome’s first river harbor . Our wor14C age determinations spanning 13500–2250 calibrated years before present , and an additional 61 age determinations on ceramic fragments incorporated in the sedimentary succession, spanning the 8th through the 2nd century BCE.The aggradational history of the Tiber River in the 30 km long valley comprised between the city of Rome and the coastal plain is strictly coupled with the post-glacial sea-level rise ,2. GeochAll necessary permits were obtained for the described study, which complied with all relevant regulations.We analyzed sediments from 11 boreholes drilled in the Forum Boarium area, between the Capitoline Hill and the Palatine Hill see . Visual The boreholes were drilled with variable depth from 15 to 53 m below the ground surface in the investigated area . ContinuVisual lithostratigraphic and in situ geotechnical (i.e. pocket penetrometer test) analyses of the cores was performed in order to describe the characteristics of the recovered sediments, including texture, granulometry, coherence, color, mineralization , presence of vegetal, faunal and anthropic material.14C and archaeological dating. Radiocarbon dating, including the treatment of the bulk sediments to retrieve plant material in FB48, was performed by Beta Analytics ; full analytical data are provided in SI dataset 1. Laura Sadori processed the palynological samples from FB38 while the 14C dates on pollen remains were provided by Center for Isotopic Research on Cultural and Environmental Heritage at the Seconda Università di Napoli . 14C dating on a piece of wood from core FB38-16 was performed by the Centre for Isotope Research at Groningen (NL).Chronological constraints were determined by a combination of All dates have been converted to calendar ages BCE and BP (2 sigma range) relative to 1950 using Cal13 dataset .When visible we collected organic and cultural remains from the identified discrete lithostratigraphic units. In addition, the sediment was sampled for wet sieving in selected cores at 10 cm interval in order to produce supplementary dating material.14C dates  was stepwise AF demagnetized in 12 steps at successive peak fields of 5, 10, 15, 20, 25, 30, 40, 50, 60, 80, 90 and 100 mT.Principal component analysis  was usedIn order to demonstrate that the ChRM record is not due to complexities and variations that arise from the magnetic mineralogy of these sediments, a set of rock magnetic analyses, based on magnetic coercivity and thermal unblocking characteristics, were conducted on representative discrete samples  is observed between 5671±32 and 4465±55 cal yr BP, consistent with the inversion in the trend of the aggradation curve occurring in the southern tract of the Tiber River Valley in Rome  and in the coastal plain [14C Hallstatt Plateau in archaeology, see 14C ages have been supplemented by dated cultural material which offers a more nuanced and reliable chronology. This has allowed us to check inconsistencies in the geochronological sequence and to recognize some apparent contradictory results due to the re-deposition of older material. Moreover, the introduction of mortar, a Roman invention of the mid-2nd century BCE [th and early 5th century BCE [For the period 700 BCE-400 BCE  that could be compared with each other. In particular, besides lateral correlation between the sedimentary successions occurring in FB43-FB47 and in FB38-FB40, analyses were aimed at checking the possible coeval age of the two lacustrine clay sections occurring at different elevations in FB38 and FB40.NRM) parameter imply that magnetite (and/or low-Ti titanomagnetite) is the primary remanence carrier. Characteristic remanence magnetization (ChRM) directions for most analyzed samples tend toward the origin of the vector component plots, with maximum angular deviation (MAD) of 3.5° on average and a range of variation between 0.6° and 18°. All cores show quite large variations of the ChRM inclination vs depth with fluctuations around the expected GAD field value of 61° for the site latitude (41°.89N). Some noisy record is attributable to the silty nature of the sediment characterized by the presence of cm-thick, fine sand layers that could have hampered the aligning influence of the geomagnetic field on ferromagnetic particles. A few samples present shallow or negative ChRM inclinations  and none of these are related to lithological variations. The ChRM inclination profile features corroborated by 14C dating, allow a correlation between cores FB47 and FB43 and between FB38 and FB40 , with gravel at the base incorporating organic matter of the substrate ;the young ages characterizing the sediments at the base of this upper sedimentary succession, immediately above the contact (FB47) or slightly above it (FB48), contrasting with the much older ages of the sediment occurring immediately below it (see FB48) or at a higher elevation (see FB38).14C age of 2800±50 cal yr BP around 1 m b.s.l. in FB48 indicates that the early stages of this new aggradational phase are characterized by a slow sedimentation rate  that causes the initial deposition of gravel within the fluvial channel, followed by deposition of clay in the alluvial plain. The 14C dates from these deposits in borehole FB39 fall within the range of the Hallstatt Plateau, ceramics indicate a more precise date around 2550 BP  for the beginning of the fast aggradation phase. In addition, building phases at the Sant’Omobono temple, whose tuff platform encountered in borehole FB 47 rests above the alluvial succession, provide key chronological upper boundary at 2430 yr BP for the end. Therefore, the archaeological evidence suggests a time span comprised between the beginning of 6th and early 5th century BCE for the deposition of the sediments comprised between 1 m b.s.l. and 5 m a.s.l. at cores 39, 43, 47, in the Forum Boarium, corresponding to lithotypes A1  and A2 .As the three The coeval, homogeneous filling of this area of the alluvial valley is consistent with the overlapping paleomagnetic signals recorded in the two portions of sediment occurring at the same elevation in core FB43 and FB47 . This seRapid re-incision of this massive package of sediments soon after its deposition is supported by the occurrence of ceramics dated 2400–2300 yr BP at elevation around 4 m a.s.l. (FB38 and FB46). In contrast, the anthropic fill at ca. 1 m b.s.l. incorporating ceramics dated ≤2100 yr BP at FB49 is problematic, and cannot be explained with the re-incision of the riverbed down to a depth below the sea-level. However, sediment compaction and/or human intervention can justify the observed low elevation of the base of anthropic fill.Finally, a new sudden sediment aggradation is evidenced around 2250 yr BP, based on the rising of a thick anthropic fill up to the previous elevation of the floodplain at around 9 m a.s.l. , and based on the contemporaneous deposition of floodplain sediments  without such a visible immediate effect on the Tiber sedimentation rate see .One possible alternative explanation to this remarkable alluvial aggradation is the co-occurrence of tectonic subsidence within the Tiber Valley, accounting for a sudden increase in accommodation space, which allows for the observed rapid sedimentary filling.In a digital elevation model (DEM) image of the area of Rome  is possiSuch feature suggests that the Tiber Island, located exactly along the continuation of the inferred fault segment, may be the result of its continued activity in early historical times. Indeed, the Tiber course seems strongly controlled by the presence of the fault in this sector, which through the lowering and the NE tilting of its hanging wall, may have generated a step acting as a natural dam to the riverbed . TectoniTectonic subsidence seems also suggested by the possible coeval age and postulated 3.2 m dislocation of the two lacustrine sections at different elevation in FB38 and FB40, as inferred by paleomagnetic analysis.Based on the geomorphological evidence described above, we have restored the original stratigraphic setting in the investigated area by hypothesizing a fault system characterized by a main NW-SE striking lineament developing in two parallel, en-echelon segments. Surface trace of these fault segments  is shown in the maps of th-7th century BCE, when a raised fluvial terrace at the foot of the Capitoline Hill overlooked the alluvial plain of the Tiber River. The shifting of the riverbed close to this sector is recorded around 2600 yr BP by the presence of a continuous layer of coarse sediments in FB39, FB48, FB47 and FB43. This is interpreted in th century BCE provides differential collapse of the western sector of the alluvial plain, for a cumulated displacement of 3.2 and 5 m, respectively  during the 6ectively . This caectively . Evidenc0 yr BP, .th and the mid-5th century the area in front of Sant'Omobono became a subsiding marshy plain, crossed by the river whose ephemeral bed was controlled by the fault scarp.The faulting process and the consequent lowering of the adjacent sector of the alluvial valley probably needs to be understood as a slow, progressive phenomenon, during which the newly created accommodation space is continuously filled by sedimentary material released by the river. The riverbed is constrained by the fault scarp along the B segment, acting as a dam and providing the condition for the origin and the stable positioning of the Tiber Island . The pro14C and archaeological age constraints on the ~4 to 7 m thick sedimentary fill  indicate that the action of faulting and the related subsidence ended in the second half of the 5th century BCE (~2430 yr BP), when a stable flooding surface was established between 5–6 m a.s.l.  BCE; [rd century may indicate abandoning of the river port and ceasing of dredging activity.Indeed, the re-incision of the alluvial plain may be explained through dredging of the fluvial channel since the mid-5 channel , this in channel . Dredginer delta  Indeed, 19) BCE; ). The do19) BCE; ,68. TherThe aggradational history of the Tiber River in central Rome is defined by three significant erosional/depositional events, which appear unrelated to the global eustatic signal and only partially related with the climatic events for the Holocene.The earliest event is characterized by a marked erosional phase around 5200 yr BP, re-excavating down to -6 m a.s.l. an early alluvial plain of the Tiber River established in Rome since ~5600 yr BP at elevation around 4 m a.s.l. Lack of information on the full thickness of the investigated sedimentary succession at Forum Boarium hinders assessment of variation in original absolute elevation of the dated erosional surface due to time-dependent sediment compaction, preventing us from discussing possible eustatic/tectonic versus climatic/sedimentological forcing of this erosive phase.th and the early 5th century BCE. However, concurrent climatic and especially anthropogenic causes are likely to contribute to the increased sediment accumulation. In this scenario, the deposition of several meters of alluvium would be indeed the result of several factors. Sediment released by increasing erosion, due to growing anthropic disturbance and landscape modifications connected to urbanization processes occurring in the 6th century BCE, further exacerbated by a shift towards wetter climate and few centuries of deforestation, would find a trap in the space created by the fault.The second event follows the progressive recovery of the floodplain up to present day sea-level, and is characterized by the sudden deposition of an ca. 6 m-thick package of sediment in one century, 2550 through 2450 yr BP. Based on the analysis of convergent geomorphological, structural and chronostratigraphic elements, corroborated by paleomagnetic investigation, we test the hypothesis that the abrupt overflooding of the river valley may be the consequence of fault displacement along a morpho-structural lineament crossing the Forum Boarium area, creating coupled increase in accommodation space and loss in capacity of transport by the river. Far from being definitive, the provided evidence accounts for the hypothesized fault displacement causing the consequent deposition of an extraordinary thick succession of sediments, the diversion of the river course, and the birth of the Tiber Island between the 6th century BCE. Urban growth in 6th century Rome is characterized by large-scale building and landscape modification projects, efforts that are well-documented and coincide with the marked increase in sedimentation in the Forum Boarium valley.Archaeological evidence demonstrates a significant increase in anthropic activity on the local landscape beginning in the late 7rd century BCE.As the city of Rome continued to expand in Republican Period, a third, less marked rapid increase in elevation of the alluvial plain occurs during the 3By providing new data on the changing hydrology of the Tiber River over the entire Holocene, this reconstruction has both scientific and historical implications for our understanding of the origins of the Eternal City and the impact of urban systems on the local and regional landscape. Additional cores, both on the Tiber Island and north of the fault displacement, are needed in order to fully support these hypotheses as well as framing them within the historical and archaeological context of the early occupation of Rome.S1 FileFigure A—Lithological/geotechnical sedimentary model for the alluvial deposits of Tiber River in Rome. See text for legend and description. Figure B—Measured magnetic parameters. Downcore variation of natural remanent magnetization at 0 mT (NRM), characteristic remanent magnetization (ChRM) declination and inclination, maximum angular deviation (MAD), magnetic susceptibility (k), anhysteretic remanent magnetization (ARM), median destructive field of the NRM (MDFnrm). See Figure C—Aggradational history of the Tiber River.14C age constraints provided by literature data  and by the present study to sediment aggradation in the Tiber Valley between Rome and the coastline. A cross-section longitudinal to the river course is reconstructed using all available borehole data , and following the 5200 yr BP erosional phase (B), until the formation of a new alluvial plain around 2800 yr BP (C), based on the reported core chronostratigraphy.(Word) (DOC)Click here for additional data file.S2 FileAll 14C ages are newly reported, except the last four (labelled SO) which have been published previously –5.(DOC)Click here for additional data file.S3 File(PDF)Click here for additional data file.S4 File(DOC)Click here for additional data file.


Moreover, hypercapnic acidosis induced pre-inspiratory discharges in the CN, HGN, SLN, and RLN. The onset of pre-inspiratory discharges in the CN, HGN, and SLN was prior to that of discharges in the RLN. These results suggest that the securing of the airway that occurs a certain time before dilation of the glottis may facilitate ventilation and improve hypercapnic acidosis.Maintaining the patency of the upper airway during breathing is of vital importance. The activity of various muscles is related to the patency of the upper airway. In the present study, we examined the respiratory motor activity in the efferent nerves innervating the upper airway muscles to determine the movements of the upper airway during respiration under normocapnic conditions (pH = 7.4) and in hypercapnic acidosis (pH = 7.2). Experiments were performed on arterially perfused decerebrate rats aged between postnatal days 21–35. We recorded the efferent nerve activity in a branch of the cervical spinal nerve innervating the infrahyoid muscles (CN), the hypoglossal nerve (HGN), the external branch of the superior laryngeal nerve (SLN), and the recurrent laryngeal nerve (RLN) with the phrenic nerve (PN). Inspiratory nerve discharges were observed in all these nerves under normocapnic conditions. The onset of inspiratory discharges in the CN and HGN was slightly prior to those in the SLN and RLN. When the CO HGN activity is exhibited in the inspiratory phase and the onset of HGN activity occurs prior to the onset of phrenic nerve (PN) activity during eupnea [Coordination of the patency of the upper airway with contraction of the diaphragm and intercostal muscles is important during breathing. The activity of various muscles is related to the patency of the upper airway, including the muscles of the tongue and lingual radix  innervated by the hypoglossal nerve (HGN), the infrahyoid muscles  innervated by the ansa cervicalis receiving fibers from the HGN or 1st–3rd cervical spinal nerves (CN) , the crig eupnea , 5. Furtg eupnea , 7. None2 [Hypercapnic acidosis is a pathological condition induced by chronically obstructive pulmonary disease, such as bronchial asthma and sleep-related breathing disorders. Pharyngeal airway obstruction during sleep is proposed to be caused by a reduction in the inspiratory activity in the genioglossus . When hy2 , 10, and2 chemosensitivity due to the inhibition of the spontaneous firing of serotonin neurons, which are known to be CO2-sensitive. Thus, in this study, we examined the respiratory motor activity in the efferent nerves innervating the upper airway muscles using in situ arterially perfused decerebrate rat preparations [in vivo mammal preparations. We also analyzed the differences in the burst onset of each nerve during eupnea and forced respiration to verify the hypothesis that the activity pattern of the upper airway muscles changes under forced expiration to promote the excretion of CO2.Anesthesia is widely acknowledged to depress the respiratory rhythm and respiratory neural discharges. Respiratory electrical activity in the cranial nerves and their innervation of the upper airway muscles has been found to be more sensitive to anesthesia than activity in the phrenic nerve , 12. Furarations , 16, enaarations  and firiarations  are simi3, 1.25 KH2PO4, 1.25 MgSO4, 2.5 CaCl2, 10 dextrose). In the cooled Ringer’s solution, the rats were decerebrated at the precollicular level. Preparations were then transferred to a recording chamber, and the descending aorta was cannulated with a double lumen catheter . Ringer’s solution containing 1.25% Ficoll  and heparin (10 unit/mL) was retrogradely perfused using a roller pump . The perfusate was continuously gassed with 5% CO2 and 95% O2 and warmed to 31–32°C (temperature measured at the point of entry into the aorta) using an in-line heater . The second lumen of the catheter was used to monitor aortic perfusion pressure. The perfusion pressure was maintained in the range of 35–50 mmHg by adjusting the flow between 25 and 38 mL/min.All experiments were performed with the approval (No. 15032) of the Institutional Animal Care and Use Committee of Showa University, which operates under Japanese Governmental Law (No. 105) for the care and use of laboratory animals. All efforts were made to minimize the suffering and number of animals used. Twenty-one Wistar rats of either sex aged between 21 and 35 days were used in this study. The weights ranged from 80 to 140 g. Preparations were prepared using a method similar to that described by Paton and St. John . The ratPreparations were paralyzed using vecuronium bromide  prior to the isolation of the peripheral nerves. Using 5 preparations, simultaneous recordings of the motor activity in the RLN, SLN, CN, and PN were obtained with bipolar suction electrodes mounted on separate 3-D micromanipulators. Each peripheral nerve on the left side was isolated, cut distally, and the proximal end was held in the suction electrodes. The PN was isolated from the pleura and cut at the distal end. The CN was identified between the left common carotid artery and sternothyroid muscle. The CN was cut proximally, 1 mm from the left common carotid artery. The SLN and RLN were identified between the trachea and the left common carotid artery. The SLN was cut in the vicinity of the trachea and the RLN was cut at the level of the thyroid gland. Using the other 5 preparations, simultaneous recordings of motor activity in the HGN, SLN, CN, and PN were obtained. The HGN was identified under the mylohyoid muscle and cut immediately before the genioglossus muscle. Using the remaining 5 and 6 preparations, motor activity in the intercostal nerve (ICN) and abdominal nerve (AbN) was recorded with CN and PN motor activity, respectively. The ICN was dissected from between the innermost intercostal muscle and internal intercostal muscle on the left at the 5th–7th thoracic spinal level (T5–T7). The AbN activity was recorded from the 1st lumbar spinal nerve (L1). All signals were amplified using a differential amplifier , band-pass filtered (1–3k Hz), and stored on a computer using an analog-to-digital converter  with version 8 of the Spike2 software (Cambridge Electronic Design). During post-hoc analysis, additional digital filtering was applied using DC remove with a time constant of 0.1 s (Spike2 software) when necessary to remove movement artifact. The nerve activity was rectified and integrated with a time constant of 0.1 s in the Spike2 software.2 and 95% O2) to a hypercapnic mixture (8% CO2 and 92% O2) for 15 min. The pH in the perfusate was altered from 7.4 to 7.2 via changes in the perfusate gas. The perfusate for hypercapnia was prepared in a separate reservoir and was gassed for at least 15 min prior to its administration.For hypercapnic stimulation, the perfusate gas composition was altered from normal carbogen . The expiratory duration was the duration of phrenic nerve activity not above the baseline noise per cycle. The integrated nerve activity in each nerve was used to obtain the peak amplitude of the discharge, onset of the discharge, and increasing duration from onset until peak PN discharge (time to peak). These values were obtained using nerve discharges during the last 1 min prior to the changes in the perfusate gas (control), 14–15 min after the perfusate changes (hypercapnia), and 14–15 min after the restoration of the perfusate (recovery). The duration of the pre-inspiratory (pre-I) discharge in the CN and ICN was measured for 5 min to calculate the correlation coefficient. In the experiments in which the AbN activity was recorded, we measured the duration of the CN pre-I discharge, AbN pre-I discharge, and respiratory cycle from 5 min before the perfusate changed to 5 min after the restoration of the perfusate. The respiratory cycle duration was defined as the time from the beginning of one phrenic nerve burst to the next.Values are presented as the mean ± standard error of the mean (SEM). Data obtained before and after changes in the perfusate gas were subjected to the Wilcoxon signed-rank test or Friedman analysis of variance (ANOVA) test. The Friedman ANOVA was followed by the Wilcoxon singed-rank test when appropriate. Differences in data between groups were analyzed using a two-way ANOVA or Kruskal-Wallis one-way ANOVA. The ANOVA was followed by a Bonferroni post-hoc multiple comparison test when appropriate. Probability values of less than 0.05 were considered significant. Statistical analyses were conducted using SPSS 17.0J  and Microsoft Excel 2011.2) to determine the activation patterns of the upper airway during respiration. In 5 preparations, the motor activities were simultaneously recorded from the CN, SLN, RLN, and PN. The integrated PN activity showed an augmenting inspiratory pattern under normocapnic conditions . Moreover, the pre-I discharges were induced prior to the PN discharges in all preparations under hypercapnic conditions . Furthermore, we recorded the nerve activity of the AbN at the 1st lumbar level in 6 preparations. Under normocapnic conditions, a distinct discharge was not observed in the AbN , the duration of the CN pre-I discharge and respiratory cycle was prolonged (P = 0.030); the difference between the respiratory duration during the last 1 min prior to changing of the perfusate and the duration during the first 0.5–1.5 min after switching approached significance . The duration of the CN pre-I discharge and respiratory cycle during the 14–15 min after changing of the perfusate was significantly longer than that during the first 0.5–1.5 min after changing of the perfusate . The duration of the AbN pre-I discharge was positively correlated with the duration of the CN pre-I discharge in hypercapnic acidosis during the 5 min after changing of the perfusate , which is similar to the PO2 level reported by Wilson et al. [The mean duration of PN discharges in our preparations was slightly longer than the discharge duration of the PN that has been suggested as a value (≤1 s) in optimally perfused preparations by Levitt et al.  and Bautvivo rat . Since tn et al. , our pren et al. .in vivo [in situ [in vivo experiments using plethysmography, Holley et al. [in situ rat preparations, many previous studies have demonstrated that hypercapnia increases the respiratory frequency [2 concentration in the perfusate from 5% to 8%, the respiratory frequency initially tended to increase; however, the frequency gradually decreased as the AbN and CN pre-I discharge appeared and grew. Similar relationships between the occurrence of the AbN pre-I discharge and slowing of respiratory rhythm have been reported by Abdala et al. [In the present study, hypercapnia increased the amplitude of PN activity and decreased inspiratory duration, consistent with the findings in many previous studies in vivo , 33 and [in situ , 34, 35.y et al.  reportedy et al.  reportedy et al.  reportedrequency , 34, 38,a et al.  and Molka et al. . Thus, mThe present study showed that hypercapnic acidosis evoked pre-I discharges in the CN, SLN, and RLN, in addition to the HGN, which has often been shown to display pre-I discharge during hypercapnia , 20, 39.2-induced hyperpnea in normal humans [The RLN includes both efferents innervating the laryngeal abductors and laryngeal adductors. Dutschmann et al.  assumed l humans  and duril humans . Such a