R0 (basic reproduction number) of 15 and the initial conditions as the reported COVID-19 infections on 17 December 2022, model simulations indicate that the Spring Festival travel can substantially increase the national peak magnitude of infection. The infection peaks arrive at most cities 1–4 days earlier as compared to those of the non-holiday time. While peak infections in certain large cities, such as Beijing and Shanghai, are decreased due to the massive migration of people to smaller cities during the pre-Spring Festival period, peak infections increase significantly in small- or medium-sized cities. For a less transmissible disease (R0 = 5), infection peaks in large cities are delayed until after the Spring Festival. Small- or medium-sized cities may experience a larger infection due to the large-scale population migration from metropolitan areas. The increased disease burden may impose considerable strain on the healthcare systems in these resource-limited areas. For a less transmissible disease, particular attention needs to be paid to outbreaks in large cities when people resume work after holidays.The large population movement during the Spring Festival travel in China can considerably accelerate the spread of epidemics, especially after the relaxation of strict control measures against COVID-19. This study aims to assess the impact of population migration in Spring Festival holiday on epidemic spread under different scenarios. Using inter-city population movement data, we construct the population flow network during the non-holiday time as well as the Spring Festival holiday. We build a large-scale metapopulation model to simulate the epidemic spread among 371 Chinese cities. We analyze the impact of Spring Festival travel on the peak timing and peak magnitude nationally and in each city. Assuming an  Human mobility can shape the transmission of epidemics at different geographical scales ,3,4,5,6.Mass vaccination can effectively reduce the incidence of severe illness caused by Omicron . With a R0 to highlight the impact of the Spring Festival travel on epidemics with various levels of disease transmissibility.We use the pre-pandemic level inter-city mobility data to simulate population movement across 371 Chinese cities after relaxing epidemic control measures. We construct a large-scale metapopulation model using the inter-city traffic data. Using COVID-19 as a motivating example, we analyze the impact of the Spring Festival travel on epidemic spread at both national and city levels. Note that we do not aim to project the actual spread of Omicron across China; instead, our objective is to use mathematical models and simulations to study the impact of the Spring Festival travel on general epidemic spread. In addition, we also run model simulations under different Using traffic flow data for epidemic simulation is an effective method . This stAccording to the official definition, the 15 days before the Spring Festival and the 25 days  after the Spring Festival, i.e., 1 February to 12 March 2018, were considered as the Spring Festival travel season. Other major official Chinese holidays include New Year’s Day (30 December 2017–1 January 2018), Lantern Festival (2 March 2018), Tomb-sweeping Festival (5 April 2018–7 April 2018), Labor Day (29 April 2018–1 May 2018), Dragon Boat Festival (16 June 2018–18 June 2018), Mid-Autumn Festival (22 September 2018–24 September 2018), and National Day (1 October 2018–7 October 2018). This study considers the dates outside of these major official Chinese holidays as the non-holiday time.We simulated the spread of epidemics during the period of non-holiday time  versus during the Spring Festival travel season .i. We define patients with symptoms severe enough to be confirmed as documented infected individuals, whereas other infections are defined as undocumented infected individuals. We define a transmission rate β for documented infected individuals. The transmission rate for undocumented individuals is reduced by multiplying a factor Z is the latency period, and D is the duration of infection. Spatial coupling within the model is represented by the daily number of people traveling from city j to city i , which is central to many epidemiologic models. Simulation of disease transmission using mathematical models is an effective method for predicting epidemic development ,20,21,22R0) is an indicator of the infectiousness or transmissibility of infectious and parasitic agents; it is defined as the average number of secondary cases generated by a case in a fully susceptible population [R0 of an infectious disease event is usually reported as a single value or low–high range; if the value of R0 is >1, the outbreak continues, and if the value of R0 is <1, the outbreak will end [R0 value for that event [The basic reproduction number μβD . We usedThis study considers only the virus transmission during the Spring Festival travel season and uses forty days of traffic flow data. It is difficult to reach steady states, and therefore we did not study the steady states of the model. Since this study focuses on the effect of Spring Festival travel on susceptible and infected individuals, the removal state individuals were not reflected in the formula. However, in order to satisfy the compartment conservation law of the model, the removal of infectious individuals was present in the actual simulation.We used model simulations to examine the impact of Spring Festival travel on the spatiotemporal transmission of epidemics. Specifically, we focused on two metrics that are crucial for policy-making and disease control: peak timing and peak magnitude for active infections. Peak timing is defined as the date with the largest number of active infections  as a proportion of the total population); peak magnitude represents the fraction of active infections at the peak among the total population.As shown in A clear asymmetric travel pattern emerged during the Spring Festival travel B. BeforeR0 = 15 and using the reported COVID-19 infection on 17 December 2022 as the initial condition  during the non-holiday time. During Spring Festival travel, most cities reach their peaks 1–4 days earlier compared to the non-holiday time and most cities without initial infections peak before day 14 as well .R0 = 15, most infections occur before the Spring Festival. Pre-Spring Festival travel brings a large number of infections from large cities to smaller cities. In addition, the migration from metropolitan areas greatly increases the susceptible pool in other cities. This results in a significantly higher peak magnitude in small- or medium-sized cities compared to the non-holiday time. In contrast, some large cities, such as Beijing, Shanghai, Guangzhou, Shenzhen, and Hangzhou, have lower peak magnitudes in the Spring Festival travel , peaks in large cities are delayed until after the Spring Festival, when a large number of susceptible and infected people return to large cities and fuel the epidemic spread. The peak magnitude is similar to or even higher than in the non-holiday time, which could stress the capacity of healthcare systems.The temporal difference in the number of infections among travelers can dynamically impact the spatial distribution of infected and susceptible populations in all cities, which further determines the variation in local peak timing and magnitude. For instance, for a less transmissible disease  for patients with severe illness. For less transmissible pathogens, we should pay attention not only to the epidemic situation in small- or medium-sized cities during the homecoming period but also to the rapid epidemic spread in large cities during the returning period after holidays.The impact of the Spring Festival travel on epidemic spread is compounded by several factors. First, population migration shapes the spatial distribution of infectious and susceptible populations across cities. The asymmetric traffic before and after the Spring Festival creates a temporal fluctuation of population present in each city, which strongly impact local transmission dynamics. Second, disease transmissibility determines the speed at which infections unfold within a population. The infection prevalence among travelers in the homecoming and returning traffic depends on the temporal growth of epidemics in each city. It is the combination of local infection dynamics and the timing of migration that collectively determines the effect on peak timing and magnitude.There are several limitations in this modeling study, both in terms of data and model. The initial conditions of epidemic simulation  have a large impact on virus transmission, and specific analyses should be conducted for each type of situation, a single model with some parameters cannot profile all results. Tencent data do not fully cover mobility across all pairs of cities, which leads to the absence of some inter-city traffic flows. The model only considers the spatial variation of travelers on a particular day but does not consider the duration of the specific trip. This study does not consider other holidays covered during the simulation and mainly focuses on the differences between the Spring Festival travel and the non-holiday time. In reality, vaccination or mask-wearing and other measures will change model parameters, which will lead to different outcomes between the simulation results and in reality. This study does not take into consideration the effects of intrinsic fluctuations and the steady states of the model. We do not pay much attention to the dynamic behavior of the epidemic model.While reducing epidemic prevention and control measures and restoring the normal life of the nation, research on various types of viruses should be maintained to reduce the harm to the economy and life from outbreaks caused by new viruses and virus mutations, and to focus on the differences between special periods and daily life to measure their impact on the epidemic to develop targeted measures.R0 > 10 on small- or medium-sized cities and the impact of viruses with R0 < 5 on large cities. This study provides a quantitative reference for Spring Festival travel epidemic prevention and control for regional governments in China.The Spring Festival is an important traditional holiday in China, and the increase in overall national population travel during the Spring Festival travel can accelerate the epidemic spread and increase the peak magnitude of infections nationwide. Different infectious viruses affect the infection rate of the population returning home before the Spring Festival and the population returning to work after the Spring Festival, and cities have asymmetries in the inflow and outflow of population before and after the Spring Festival; these affect the peak time and peak magnitude of cities, with particular attention to the impact of highly infectious viruses with


The professional identity of the ‘genetic counsellor’ first took shape in the UK in the early 1990s, when the University of Manchester established the country’s first masters-level training course. Postwar, genetic counselling had been carried out by  clinical geneticists, who, alongside their research, clinical and field-building activities, met patients and families to discuss inherited conditions and risk estimates, and who sometimes advised parents whether to attempt or continue pregnancies. By contrast, the new cohort of students in Manchester in the 1990s were not medically trained, were mostly women, and were schooled in the psychological and social consequences of genetic testing and diagnosis, as well as methods for the care, support and emotional management of patients and families. This was a significant change both in the practices of ‘genetic counselling’ and who was expected to practise it. Focusing on a small section of this history, between 1980 and 1995, this paper describes some of the historical threads that contributed to this change. It charts the early work of genetic nurses and social workers, who in the 1980s carved out distinctive roles within National Health Service genetics centres. It describes the separate, specialist provision developed by sickle cell and thalassaemia counsellors, who developed new approaches in dialogue with racialised and underserved patient communities. It examines growing interest in the late 1980s and early 1990s in the tacit social and cultural conditions of genetic counselling encounters, and how this cohered with attention from disability scholars, psychologists and social scientists. By describing these historical contributions, this paper explores how the intersecting gendered, racialised and disciplinary politics of clinical genetics shaped the new professional role of the ‘genetic counsellor’. Today, clients usually encounter genetic counselling during pregnancy, cancer treatment, or in the course of paediatric care, although increasingly they might be given referrals from other hospital departments.The identity of ‘the genetic counsellor’ consolidated in the UK in the early 1990s, when the University of Manchester established the country’s first degree-level training course. The course was the culmination of a remarkable shift in genetic counselling practices over several decades. For several decades postwar, genetic counselling in the UK had been carried out by  clinical geneticists, who, alongside their clinical, research and field-building activities, met patients and families to discuss inheritance and risk, and who sometimes gave advice about reproductive decisions, before or during pregnancy eg, . By contHere I trace in particular the growing significance of care, emotional management and psychotherapeutic interactions in genetic counselling.The 1980s was also an important decade for the separate provision given to patients and families affected by the serious and unpredictable heritable haemoglobin conditions sickle cell disease (SCD) and thalassaemia. These conditions have tended to affect people with recent ancestry in Africa, the Mediterranean and South Asia—groups that, in the UK, often also experienced considerable racism and neglect within the postwar welfare system. Ignorance, racism and lack of training within the NHS meant that patients suffering from these conditions experienced inadequate or no healthcare during the postwar decades .6 As a cGenetic nurses and social workers, and sickle cell and thalassaemia counsellors, all contributed to a growing interest in the late 1980s in the tacit social and cultural conditions of genetic counselling—the topic of section three. Many practitioners perceived that the social and cultural experiences of clients and professionals could profoundly affect the kinds of information that could be communicated, how that information was understood, and what kind of support was possible. Genetic information did not mean the same thing to all people: the way in which it was assimilated and acted on varied with social, cultural, gendered, ‘disablised’ and ‘racialized’ identities and experiences.Thus, this paper attempts to trace the history of a specific, ephemeral, talking-based, medical encounter.What roles did genetic nurses and genetic social workers have in the NHS? By 1980, clinical genetics in the NHS had been expanding for three decades, and had grown from a few ad hoc units run by clinicians with special interests in genetics, to a total of 16 genetic centres across the ‘regions’ of the UK, led by trained clinical geneticists . A key mWho were these ‘auxiliary’ workers and what did they do? The earliest such appointment was apparently made in 1959 by Cedric Carter, who ran genetics clinics at Great Ormond Street Hospital and a Medical Research Council unit at the Institute of Child Health. Carter recruited Kathleen Evans, a medical social worker at the hospital, who prepared information on clients and their families prior to clinics, who talked to the families before they met with Carter, and who accompanied Carter in the clinics , 29.12 OWe know more about the responsibilities of the first so-called ‘genetics field worker’ in the UK, a woman named Edith Quinn who was a senior registered nurse when appointed in 1971 by clinical geneticist Rodney Harris to work in the Manchester Department of Medical Genetics. Two years later, he and Quinn coauthored a memo explaining her multiple responsibilities in the department’s research and clinical work . Like EvPractical Genetic Counselling (1981), Cardiff-based clinical geneticist Peter Harper praised the important work by ‘auxiliary workers’ in clinical genetics, noting that they were uniquely able to ‘detect problems that the family have not spoken about, and ensure that they have actually understood what the person giving genetic counselling thinks they have’. But Harper saw these activities as distinct from ‘genetic counselling’, which was the preserve of clinical geneticists.Others evidently agreed with Harris, and over the next decade, centres around the country brought in nurses and social workers to take on similar roles as those of Evans and Quinn. As their work gradually began to be acknowledged in print, colleagues described the expertise of nurses and social workers in gauging the attitudes and responses of families, and offering appropriate support. In the textbook Harper was not alone. In services across the country, nurses performed supporting roles to clinical geneticists, who had authority over clinics, and made decisions regarding the information that needed to be collected from which clients and families.During the 1980s, published accounts began alluding to the ‘psychological’ aspects of nurses’ work. A Clinical Genetics Society report published in 1982 explained that nursing staff hired to the genetics services should have experience in community medicine, paediatrics and obstetrics. People with this training, it explained, ‘will be aware of the many problems resulting from genetic disease in families, and should be sensitive to the psychological implications for the affected individuals and their relatives’ , 11. Loog-nas-wa). Twenty people attended the first meeting, held in September 1980, and they travelled from the genetic services in Cardiff, Edinburgh, Belfast, Nottingham, Manchester, Preston and Cheshire. Eleven were nurses or health visitors, eight were social workers and one was a psychologist  the nursing care, of patients in the hospital. Also, with my health visitor hat on I was concerned to support the families. No, it wasn't selling! everything’ , and later reflected on the significance of cultural identities in the centre. She recalled, ‘[genetic counselling] is a very emotive subject; it’s one that affects reproductive choices, there are cultural issues, religious issues … and I didn’t think it was adequate trying to interpret through a younger member of the family, a husband’ . Most of its essays were written by US-based psychiatrists and psychologists, who offered ‘practical guidance to important psychological problems involved in genetic counselling and the skills required to tackle them’.‘content-oriented counselling’ and towards ‘person-oriented counselling’   , 4. The mphasis) , now a nmphasis) .Psychological Aspects was reviewed largely positively in the USA and UK , an organisation established by a group of social workers in Groningen in the Netherlands . AuthorsBy the early 1990s, some NHS regional genetic centres were engaging explicitly and practically with these innovations. For example, unusually, the Yorkshire Regional Genetics Service in Leeds had on its team a psychotherapist, Christina Oliver, who had trained as a midwife. Anna Middleton, who was associated with the Leeds centre from 1992, recalls the profound influence that Oliver had on the practices of genetic counsellors on the team, and that it was one of the first units in the country to introduce ‘supervision’ for genetic counsellors. For the Leeds team, supervision—by then an established practice in psychotherapy—meant confidential meetings between a counsellor and a colleague in order to reflect on the emotional encounters that they were dealing with, especially with respect to difficult cases.The 1990s was the decade of the Human Genome Project and ‘the new genetics’, which attracted attention from disability scholars and activists deeply concerned with the potential for expanded genetic testing to reify a biological determinist concept of disability as medical incapacity.American Journal of Human Genetics in 1998, Middleton argued that employing deaf genetic counsellors would improve services for deaf people, because of better language communication and because of their cultural awareness: ‘A deaf genetic counsellor would be fluent in sign language and would have a cultural awareness as well as first-hand knowledge of issues relevant to deaf people’ . people’ , 1179. C people’ , 1179.I have described here how, in the mid-to-late 1980s and 1990s, UK healthcare professionals began to reinvent genetic counselling as a psychological, emotional and social encounter. Genetic nurses, social workers and haemoglobinopathy counsellors, in collaboration with clinical geneticists, sociologists, psychologists and other social scientists, created forums for exchanging ideas and practices, a flourishing vocabulary, and an increasingly varied body of research on the social and psychological responses of clients and families to clinical genetic information. It was this ‘psychosocial’ emphasis that would become a central feature of the new genetic counselling master’s course first offered in Manchester in 1992.As we have seen so far, the 1980s was formative for the UK’s development of practices delivered in regional genetics centres by genetic nurses, and within the haemoglobinopathy services by sickle cell counsellors and thalassaemia counsellors. During the early part of that decade, those ‘non-medical’ counselling roles were still relatively invisible—in the sense that they did not involve nationally organised training and they were rarely mentioned in journals or textbooks. There were also few opportunities for formal training, which tended to happen ‘in-house’ within individual institutions. One exception was a week-long training course for haemoglobinopathy counsellors run by Elizabeth Anionwu at the Brent Sickle Cell and Thalassaemia Clinic . AnotherBorn in the USA, Kerzin-Storrar had been studying in San Diego for a first degree in biology in the mid-1970s when she heard about the master’s course in Genetic Counselling at Berkeley, California. The first such graduate degree had been established at Sarah Lawrence College in New York in 1969; a handful of others followed suit and the Berkeley course was set up in 1973.The orientation of the Berkeley degree had a profound impact on Kerzin-Storrar’s own vision and practice, and on her establishment of the Manchester masters in genetic counselling in the early 1990s.Also teaching on the course was Elizabeth Anionwu, by now Lecturer in Community Genetics at the Institute of Child Health at UCL. Anionwu schooled students on haemoglobin-related conditions, but also on issues of ‘ethnicity’ and counselling interactions (discussed in the previous section). Recalling Anionwu’s insights into social identity and counselling, Kerzin-Storrar commented, ‘frankly in those early days that would have been the only…thing in the entire course that actually addressed [issues of culture and identity] directly’ . AnionwuLike in Berkeley, the Manchester degree put students on placements in social care, educational or health settings, which they would do with supervision, to introduce them to the experience of reflective practice . OrganisThis alignment work happened in concert with another institutional change. In 1995, the GNSWA voted to change its name to the Association of Genetic Nurses and Counsellors (the AGNC); a letter circulated to services around the country explained, ‘it was felt that this [name] best describes the focus of most of the membership’.Practical Genetic Counselling (1993) explained that, ‘genetic counselling should preferably be undertaken by people who are medically trained, largely for the reason that it is quite impossible to separate the actual counselling from the associated aspects of clinical diagnosis’  influenced by developments on the US West Coast. The work of Elizabeth Anionwu and her colleagues in Brent was modelled on the broad support given by sickle cell counsellors in Los Angeles and Oakland—who also influenced growing interest in and attention to notions of culture and identity in counselling encounters. Meanwhile, Lauren Kerzin-Storrar brought to the Manchester degree several features of the Berkeley genetic counselling course, including an emphasis on the value of community placements, on psychological theories and issues, and on reflexive practice. Although the UK’s first master’s courses in genetic counselling lagged the US by over two decades, this perceived ‘delay’ does more to reveal some of the distinctive features of clinical genetics in the NHS. For example, in contrast to the US landscape of private healthcare, the context of state-run, nationwide, regionally organised health services—characterised by cooperation within teams and attempts to standardise services between regions—likely slowed changes to the professional identities of nurses, counsellors and social workers. The ‘team’ character of regional genetics centres already created the conditions for a local diversity of roles, training and expertise, which encouraged but also absorbed change. It is also likely that the NHS administrative structure tempered the pace of change compared with the USA. For example, Kerzin-Storrar noted that for many years her hospital Chief Executive, who supported her original appointment, did not know what to pay her, because she did not fit into an existing salary scale.Within the NHS clinical genetics services, we have seen the growing significance of the emotional labour of genetic counselling. The emotional management of clients and families by ‘auxiliary workers’ was largely unacknowledged in print until the late 1980s, perhaps both cause and consequence of this work being done by women.and with genetic nurses and social workers to carve out their new roles and expertise.Perhaps because of the commitment to team working within the NHS genetic services, the history of genetic counselling in the UK draws attention to a great deal of careful negotiation and management of roles and identities across the system of clinical genetics. Devon Stillwell, in her account of genetic counselling in the US, draws on Andrew Abbott’s concept of an interrelated ‘system of professions’—in which the development of one profession is shaped by competition with others it is in contact with—to describe the complicated gender culture that structured genetic counselling . StillweThe dynamics of the relationships between UK genetic nurses and counsellors, on the one hand, and clinical geneticists, on the other, was complex and varied. Some former nurses described the paternalistic attitudes of clinical geneticists and other specialists. Others chose to emphasise the extensive support and autonomy they were given by the clinicians they worked with. Genetic nurse Heather Skirton explained, ‘I always used to think how it happened in this country was a few medical geneticists started saying, ‘my nurse can do more than just change the bedsheet on the examination table and make my coffee’’. She reflected on how those relationships likely affected the trajectory and pace of change of particular services . The conI have described how the expanding autonomy of genetic nurses and the construction of the genetic counsellor developed in concert with a turn to the ‘psychosocial’ in clinical genetics. We might understand this turn as part of a conscious distancing from ‘eugenics’, a shift that was then itself complicated by debates in the late 1980s and early 1990s about ‘non-directive counselling’.This history is ongoing. As genomics becomes ever more visible within UK healthcare, genetic counsellors continue to carve out roles as essential mediators and interpreters of such data.


The present study reports the population structure, genetic admixture and phylogeography of cattle breeds of Sri Lanka viz. Batu Harak, Thawalam and White cattle. Moderately high level of genetic diversity was observed in all the three Sri Lankan zebu cattle breeds. Estimates of inbreeding for Thawalam and White cattle breeds were relatively high with 6.1% and 7.2% respectively. Genetic differentiation of Sri Lankan Zebu (Batu Harak and White cattle) was lowest with Red Sindhi among Indus Valley Zebu while it was lowest with Hallikar among the South Indian cattle. Global F statistics showed 6.5% differences among all the investigated Zebu cattle breeds and 1.9% differences among Sri Lankan Zebu breeds. The Sri Lankan Zebu cattle breeds showed strong genetic relationships with Hallikar cattle, an ancient breed considered to be ancestor for most of the Mysore type draught cattle breeds of South India. Genetic admixture analysis revealed high levels of breed purity in Lanka White cattle with >97% Zebu ancestry. However, significant taurine admixture was observed in Batu Harak and Thawalam cattle. Two major Zebu haplogroups, I1 and I2 were observed in Sri Lankan Zebu with the former predominating the later in all the three breeds. A total of 112 haplotypes were observed in the studied breeds, of which 50 haplotypes were found in Sri Lankan Zebu cattle. Mismatch analysis revealed unimodal distribution in all the three breeds indicating population expansion. The sum of squared deviations (SSD) and raggedness index were non-significant in both the lineages of all the three breeds except for I1 lineage of Thawalam cattle (P<0.01) and I2 lineage of Batu Harak cattle (P<0.05). The results of neutrality tests revealed negative Tajima’s D values for both the lineages of Batu Harak (P>0.05) and White cattle (P>0.05) indicating an excess of low frequency polymorphisms and demographic expansion. Genetic dilution of native Zebu cattle germplasm observed in the study is a cause for concern. Hence, it is imperative that national breeding organizations consider establishing conservation units for the three native cattle breeds to maintain breed purity and initiate genetic improvement programs. Bos indicus), exotic Zebu, commercial taurine (Bos taurus) and various crosses of these three types. Historically, cattle rearing in Sri Lanka dates back to 543 BC with the arrival of “Indo Aryans” and since then, periodic introduction of various cattle breeds has taken place through time [Cattle represent a major proportion of livestock population in Sri Lanka. Cattle genetic resources in Sri Lanka comprise indigenous Zebu  and Kangayam (51) generated in a previous study  was inclTwenty seven FAO recommended microsatellite markers  with for5’ AGGACAACCAGTCGAACACC 3’) and BTMTD1-R 5’ (GTGCCTTGCTTTGGGTTAAG 3’) as reported elsewhere [The mitochondrial D-Loop region was amplified using the primers BTMTD1-F . The extent of population sub-structuring was further explored using STRUCTURE [To perform data quality control, the microsatellite genotypes were pruned for (i) missingness (ii) null alleles and (iii) deviations from selective neutrality. Individuals with >20% missing data (missing five or more genotypes out of 27 investigated loci) were excluded from the analysis. The presence of null alleles in the dataset was checked using MicroChecker version 2.2.3 . The neuersion 1 . Basic dersion 1 . Pairwisersion 1 . F statiersion 1  was calcersion 1 . AnalysiTRUCTURE  with theTRUCTURE . The optTRUCTURE .ST among studied cattle populations were calculated using ARLEQUIN 3.5.3 [ST derived from mtDNA haplotype frequency were utilized to perform non-metric multidimensional scaling (MDS) analysis using SPSS version 13.0. MEGA version 6.0 [S were conducted using ARLEQUIN 3.5.3. Median Joining (MJ) network of Sri Lankan cattle haplotypes was reconstructed using NETWORK 4.5.1.2 [Mitochondrial DNA sequences were checked and edited by using Codon Code Aligner version 3.7.1. . A total of 212 sequences generated in the present study was deposited to NCBI-GenBank (accession nos. MZ501954-MZ502165). Additionally, 87 mtDNA sequences from two South Indian cattle breeds, Hallikar  and Kangayam  were included for comparative analysis . HaplotyIN 3.5.3 . PairwisIS) among all the investigated cattle breeds was 0.054±0.016. Among Zebu cattle alone, the overall FIS was 0.063±0.020 and ranged from -0.053±0.023 (TGLA126) to 0.415±0.053 (HEL5) across different loci  deviated significantly (P<0.05). Among the Sri Lankan native cattle 17 breed x loci combinations (20.99%) deviated significantly (P<0.05) due to heterozygosity deficit. The percent HWE deviation observed in the study is comparable to Brazilian (19.5% breed x locus combinations ) and Indelection . Both thST was estimated at 0.145±0.010 indicating 14.5% of the total variation being explained by between breed differences and 85.5% of the variation due to within breed differences. The between breed differences was estimated at 6.5% when all the Zebu breeds were considered while it was only 1.9% when Sri Lankan Zebu  alone were considered. The pairwise FST estimates among Zebu cattle breeds ranged from 0.015  to 0.085 (LTM-IKA)  . Geneticu cattle . The Neiu cattle . Genetiche model . The Tuche model . The MDSST), among group variance was only 0.86% (P>0.05). However, when Hallikar cattle alone (South India) was grouped with Sri Lankan Zebu, among group variance was 2.54% and significant (P<0.01). The results clearly indicated strong genetic relationship of Sri Lankan Zebu cattle with Hallikar, an ancestral Mysore type South Indian breed known for its excellent draught characteristics. Bayesian clustering of individuals without prior population information was performed to assess the cryptic genetic structure and degree of admixture among the investigated cattle breeds. Among K = 2 to 14, our dataset was best described with K = 2 genetic clusters, at which the second order rate of change in LnP(D) was maximum  was performed to further assess the underlying genetic relationship among the investigated Zebu cattle populations and to test and validate the results of phylogeny and MDS analysis. When no grouping of Zebu cattle was assumed, AMOVA revealed 93.93% of total genetic variation was due to differences among individuals within breeds and only 6.07% was accounted for by differences between breeds . When Ze maximum . When K  maximum . When K  maximum .The haplogroup classification and diversity of Sri Lankan Zebu cattle are presented in ST among Zebu cattle from Sri Lanka, Indus Valley and South India was estimated utilizing the frequency of mtDNA haplotypes  and 0.202  while I2 haplotypes showed pairwise FST varying between zero  and 0.178 . Among the Sri Lankan Zebu breeds, Batu Harak showed low FSTs with Tharparkar (FST = 0.030) and Red Sindhi (FST = 0.039) among I1 haplogroup while it showed lowest FST with Sahiwal (FST = 0.021) among I2 haplogroup. Lanka White cattle showed low FSTs with Sahiwal (FST = 0.030) and Hallikar (FST = 0.050) among I1 haplogroup while it showed low FSTs with Red Sindhi (FST = 0.020) and Hallikar (FST = 0.053) among I2 haplogroup. Interestingly, I1 haplogroup of Thawalam cattle showed high FSTs not only with Indus Valley and South Indian cattle (FST = 0.108–0.199) but also with other Sri Lankan Zebu (FST = 0.168–0.202) breeds. However, among I2 haplogroup, Thawalam cattle showed low FSTs with Sahiwal (FST = 0.020) and Red Sindhi (FST = 0.039).Phylogenetic analysis was performed on all the 112 unique haplotypes along with reference sequences of each lineage, the results of which is presented in plotypes . AnalysiThe demographic history of Sri Lankan Zebu cattle was explored by assessing mismatch distribution of nucleotide differences among haplotypes within I1 and I2 maternal lineages . All theo, Ho and He. The extant population of Batu Harak is believed to have resulted from extensive outcrossing of original Sri Lankan Zebu cattle with various Indo-Pakistan Zebu cattle brought to Sri Lanka [IS can be associated with heterogeneity of herds sampled within each breed, possibly resulting in Wahlund effect [Our study presents the first comprehensive analysis of diversity, genetic and phylogeographic relationship of island cattle of Sri Lanka with Zebu cattle of Indus Valley and South Indian origin. The existing cattle production system of Sri Lanka comprises of at least five categories of breed types that include native Zebu cattle, exotic Zebu, native Zebu x exotic Zebu crosses, exotic taurine, and native Zebu x exotic taurine crosses. The distribution of cattle breed types varies with the agro-ecological zones with Zebu and Zebu crosses dominating the dry zone while purebred exotic taurine and native Zebu x exotic taurine crosses dominating the wet zone of the country. The intermediary zone consisted of both Zebu x taurine as well as native x exotic Zebu crosses . Variousri Lanka , 43. Thid effect . AssortaThe present-day Batu Harak cattle are believed to represent the descendants of crosses between original archaic “Sri Lankan Zebu cattle” and various Indus Valley (Indo-Pakistan) Zebu breeds , 4. The Bos taurus breeds were brought into Sri Lanka and since then crossbreeding is used as one of the methods for genetic improvement of milk production in the country. However, indiscriminate breeding of local cattle using semen of taurine and crossbred bulls resulted in dilution of native germplasm. Bayesian clustering analysis was performed to assess the taurine admixture in Sri Lankan Zebu cattle using purebred Jersey, Ayrshire and Holstein-Friesian as reference genotypes. The proportion of membership coefficient obtained for individual animals in the inferred Zebu and taurine clusters were utilized to estimate taurine admixture and purity of the investigated Zebu cattle breeds [Historically, with the arrival of Europeans, temperate e breeds . The rese breeds , 46. It e breeds . Furthere breeds  recommenThe mtDNA maternal lineages of indigenous Sri Lankan Zebu cattle belonged to I1 and I2 haplogroups typical to other South Asian Zebu cattle , 47. TheThe present study revealed moderately high levels of genetic diversity in Sri Lankan Zebu cattle breeds. The estimates of inbreeding were relatively high in Thawalam and White cattle breeds. The Sri Lankan Zebu cattle breeds showed strong genetic relationships with Hallikar cattle, an ancient breed considered to be ancestor for most of the Mysore type draught cattle breeds of South India. Recent introgressions of Indus Valley Zebu like Red Sindhi, Tharparkar and Sahiwal was also evident in Sri Lankan Zebu, especially in Batu Harak and Thawalam cattle breeds. mtDNA analysis showed two maternal lineages, I1 and I2 in Sri Lankan Zebu cattle with relatively high level of haplotype diversity in Batu Harak cattle. Genetic admixture analysis revealed high levels of breed purity in Lanka White cattle while significant taurine admixture was observed in Batu Harak and Thawalam cattle. Genetic dilution of native Zebu germplasm is a cause for concern from biodiversity and conservation standpoint. It is recommended that national breeding organizations consider establishing conservation units for the three native cattle breeds to maintain breed purity and initiate genetic improvement programs. Considering the superior adaptability characteristics of these animals, such an initiative will be beneficial in the long run to develop locally adapted dual-purpose cattle for milk and meat production in Sri Lanka.S1 File(DOCX)Click here for additional data file.S2 FileFST outlier approach using LOSITAN under the assumption of infinite allele mutation model.Selection detection based on the (DOCX)Click here for additional data file.S3 File2005) (a) Mean L (K) over 20 runs for each K value of 1 to 14 (b) Distribution of ΔK with the modal value (K = 2) indicating the true K or the uppermost level of structure.(DOCX)Click here for additional data file.S4 File(DOCX)Click here for additional data file.S5 File(DOCX)Click here for additional data file.


Pediatric H3K27M-mutant diffuse midline gliomas (DMGs), including those formerly classified as diffuse intrinsic pontine gliomas (DIPG), are uniformly lethal CNS malignancies . Childreex vivo experiments, they are often unable to effectively cross the cell membrane due to their large size and charge. Smaller, cell permeable STAT3 inhibitors often utilize covalent binding mechanisms to enhance target binding affinity and inhibition  proteins are activated by extracellular signals such as growth factors and cytokines; these proteins are implicated in survival, proliferation, and differentiation at baseline . STAT3, hibition  11]..ex vivo in vitro and induce a robust restoration of H3K27 trimethylation (H3K27me3) expression which is normally absent in DMG tumors. Finally, the STAT3 pathway inhibitor WP1066, which utilizes a reversable-covalent mechanism to enhance target binding and inhibition [As detailed in our recently published manuscript, STAT3 is a biologically relevant therapeutic target in H3K27M-mutant diffuse midline glioma , our labhibition , increasSuccessful translation of STAT3 inhibitors to the clinic, both in DMG and in other cancers, will primarily require management of off-target effects and associated toxicity. Pathway inhibitors of STAT3 are currently in clinical trials for multiple CNS malignancies. A recent Phase I clinical trial of WP1066 in recurrent GBM found that this inhibitor is able to reduce STAT3 phosphorylation in peripheral blood with a 1 mg/kg dose . In addi


Gluten content labels inform food choice and people practicing a gluten-free diet rely upon them to avoid illness. The regulations differ between jurisdictions, especially concerning fermented foodstuffs such as beer. Gluten abundance is typically measured using ELISAs, which have come into question when testing fermented or hydrolysed foodstuffs such as beer. Mass spectrometry can be used to directly identify gluten peptides and reveal false negatives recorded by ELISA. In this survey of gluten in control and gluten-free beers, gluten protein fragments that contain known immunogenic epitopes were detected using liquid chromatography-mass spectrometry in multiple beers that claim to be gluten-free and have sufficiently low gluten content, as measured by ELISA, to qualify as being gluten-free in some jurisdictions. In fact, several purportedly gluten-free beers showed equivalent or higher hordein content than some of the untreated, control beers. The shortcomings of ELISAs for beer gluten testing are summarised, the mismatch between ELISA and mass spectrometry results are explored, and the suitability of existing regulations as they pertain to the gluten content in fermented foods in different jurisdictions are discussed. Gluten proteins consist of prolamins found in wheat, barley, rye, and oats . These pFood safety labelling regulations vary across jurisdictions, though the Codex Alimentarius represents a consensus of regulatory standards . It recoThe sandwich enzyme-linked immunosorbent assay (ELISA) is the official method for gluten determination for both the Codex Alimentarius and AOAC International (US). This requires the target antigen to have two or more antibody-binding epitopes per molecule. However, in fermented or heated food products wherein gluten proteins may be digested or fragmented to peptides and thus may only contain a single epitope per molecule, a competitive ELISA is more appropriate . There aNotably, the R5 antibody does not accurately quantify barley prolamins or hydrolysed gluten . While cBeer production involves multiple steps where prolamins can be digested, degraded, or sequestered, including malting, mashing, filtering, fermenting, and stabilisation . RemainiThe technicalities involved in beer production, and then measuring and regulating gluten protein content in beer is a topic of research. The USA’s FDA has recently issued a ruling on the gluten-free labelling of fermented foods such as beer that recognises this difficulty . They ruReagents were sourced from Sigma-Aldrich , solvents from ChemSupply , and enzymes used for digestion (trypsin and chymotrypsin) from Promega . The Ridascreen R5 competitive ELISA kits were purchased from R-Biopharm Australia  and were performed according to manufacturers’ instructions. A range of beers (single bottles) were selected that included different types of malt and brewing styles, as well as regular (control), low, and reduced-gluten labelling. They were chosen based on their advertised ingredients  and gluten status claims and purchased from international commercial vendors in 2020. Two of the gluten-reduced beers were selected based on anecdotal evidence by consumers of symptoms induced by consumption. Five regular beers that had been previously confirmed to have a range of gluten content  were usen = 4 technical replicates) using either trypsin or chymotrypsin, aliquots of degassed beer (50 μL) were diluted in 50 μL of 50 mM ammonium bicarbonate containing 1 mM CaCl2, pH 8.5. To the beer, 10 μL of 50 mM dithiothreitol was then added, and the samples were incubated for 30 min at 60 °C. The samples were cooled, 10 μL of 100 mM iodoacetamide was added, and the samples were then incubated at 20 °C for 20 min in the dark. To each solution, 10 μL of either trypsin or chymotrypsin (1 μg/μL) was then added and the samples were incubated at 37 °C for 16 hours. To quench the digestion, 50 μL of 1% formic acid (FA) was added, and the samples were frozen at −20 °C until analysis. To digest whole beers  coupled to an Ekspert nanoLC415 . Aliquots measuring 5 μL were loaded onto a ChromXP C18 trap column , using 10 μL/min 0.1% FA for 5 min. Peptides were chromatographically separated on a ChromXP C18  column at a flow rate of 5 μL/min. Mobile phases A  and B  varied in a linear gradient from 5% to 45% solvent B over 40 min, then 45–90% B over 5 min, a 5 min hold at 90% B, return to 5% B over 1 min, and 14 min of re-equilibration. The HPLC eluent was directed onto a DuoSpray ion source with heated interface set to 100 °C, ion spray voltage to 5500 V, curtain gas to 20 psi, and ion source gas 1 and 2 set to 15 and 20 psi, respectively. Data were acquired in information-dependent acquisition mode, where a time-of-flight MS spectrum was recorded over mass range m/z 350–1800 with 0.25 s accumulation time. The 30 most intense precursor ions that exceeded 200 counts per second, with charge state between 2–5, were selected for tandem mass spectrometry in 30 tandem MS scans . Dynamic ion exclusion was used to avoid selecting a precursor ion after one occurrence within 8 s, with ions having a mass tolerance 50 ppm and peaks within 6 Da of the precursor being excluded. Raw data were searched using ProteinPilot software v5.0.3 (SCIEX) against a database consisting of all Poaceae entries available on UniProtKB (accessed on 21 February 2023), concatenated with common repository of adventitious proteins (thegpm.org/crap). This identified tryptic and chymotryptic peptides detected in the samples. Search results are reported at 1% global false discovery rate as determined by an algorithm incorporated into ProteinPilot .Samples (10 μL) were injected onto a UHPLC system  directly coupled to a QTRAP 6500 mass spectrometer (SCIEX) and peptides from barley hordeins and wheat glutenins and gliadins were targeted using multiple reaction monitoring (MRM) analyses to examine the LG beers and compare gluten content to control beers (C1-C5). The MRM assay cycle time was 0.3 s, and transitions were scheduled to be ± 40 s of their observed retention time. MRM data were analysed using Skyline v.21.2.0.425 , where ehttps://software.broadinstitute.org/morpheus/). Wheat peptide MRM data was similarly exported from Skyline to GraphPad where mean ± SD were plotted for each beer directly. Peptide peak area was exported from Skyline into Excel for targeted tryptic and chymotryptic assays. The mean peak area and standard deviation (SD) of each protein class  was determined across the four technical replicates for each beer and graphs were generated in GraphPad Prism . Heatmaps of peptide abundance were visualised and annotated using the web-based software Morpheus , updated 2020 ,48, wereDegassed beers were measured using an R5 competitive ELISA  according to the manufacturers’ instructions. As listed in LG beers were subjected to discovery proteomics using data-dependent LC-MS acquisition, which detected tryptic and chymotryptic peptides separately. The high sequence homology of gluten proteins means that different proteins include the same peptides in their sequence, and multiple proteins may be present in each protein group in bottom-up proteomics . The majThere were 2091 distinct peptides detected across all LG beer tryptic digests, which identified at least 199 protein groups (excluding cRAP proteins). The top identified proteins include serpins, lipid transfer proteins (LTPs), and alpha-amylase/trypsin inhibitors, as well as B-, D- and γ-hordeins . In chymotryptic digests of LG beers, there were 787 distinct peptides and 110 protein groups detected. Serpins, D-, B-, and γ-hordeins, LTPs, amylase inhibitors, and trypsin inhibitors were seen in the top 10 proteins, as is typical of beer  Table S. To determine how many potentially immunogenic proteins are detectable in the LG beers, the proteins detected in each beer were searched for known immunogenic epitopes. The protein sequences in each protein group were searched for R5 epitopes , HLA-DQ Proteins with immunogenic epitopes were detected in all LG beers except for LG9 . While tImmunogenic epitopes  in the intact protein may be destroyed in the tested sample, either from hydrolysis during the beer production process , or intentionally during sample preparation. However, both enzymes used in sample preparation (trypsin and chymotrypsin) are present in the human digestive system, so any beer proteins consumed would be expected to be cleaved by the same enzymes in vivo. Peptides that are cleaved but retain 9-amino acid core epitope regions will exhibit immunogenic properties and are potentially clinically relevant . Figure MRM assays were used to quantify gluten-derived peptides from hordeins and avenin-like proteins (ALPs) in trypsin- and chymotrypsin-digested samples that yielded high responses. MRM allows for quantitative and more sensitive measurements of peptides than DDA-MS. Of the peptides monitored in ALP protein levels were generally higher in control than in LG beers and were seen in all nine LG beers. Peptide QQQQQGQSFVQPQQQVPVEITR was seen in LG3 and LG9 at higher levels than in control beers, and was the only ALP detected in Finnish beer LG9. Members of the B-hordein family share high sequence homology, meaning that their peptides (both tryptic and chymotryptic) are markers of multiple B-hordein isoforms. While overall B-hordein content is more abundant in control than in LG beers, selected peptides were more abundant in LG beers than in control . Two pepC-hordeins have over 20 isoforms per genome , but areD-hordein presents as a single gene in barley ,54, encoBarley γ-hordeins have three isoforms  and are potential triggers of CD, but are less abundant than the B- and C-hordeins . Some vaThe relative abundances (as measured by the peptide peak area) of the gluten protein peptides in The differences in net hordein protein peak area between beers is evident in both tryptic and chymotryptic digests . In mostSince wheat-specific proteins were observed in most LG beers during discovery proteomics, the beers were subjected to MRM analysis targeting nine wheat-specific peptides that ionise well to test for potential contamination . Wheat-sGluten proteins were detected and measured in nine LG and five control beers by ELISA and targeted LC-MS. While the relative gluten protein abundance varies greatly between beers, there were differences within control and LG groups, between the types of hordeins monitored, and between the measurement method , Table 1Gluten-free labelling regulations are traditionally based on antibody ELISA measurements, such as those shown for the competitive R5 ELISA assay results presented in Epitope masking by phenolic compounds can hinder the interaction between anti-bodies and epitopes. Polyphenols extracted from cranberry or apple inhibit signal from anti-gliadin antibodies in dot blot assays, demonstrating a reversible and dose-dependent interaction between certain polyphenols and gliadins mediated by epitope masking and potentially insoluble protein-polyphenol complexes . SimilarThere is evidence for low levels of wheat content based on four wheat peptides from three wheat HMW glutenin isoforms  in six of the LG beers: LG1, LG2, LG5, LG6, LG7, and LG8 . NotablyC-hordein peptides were detected in most of the LG beers, except for LG5, LG6, and LG9. The Finnish beers, LG7 and LG8, showed elevated levels of C- hordeins, and LG2 contained two C-hordein peptides that were not present in any other LG beer. This class of hordeins is particularly important, as several contain epitopes that are highly immunogenic to CD patients . Figure The presence of immunogenic epitopes in LG beers illustrates the technical hurdles to quantifying gluten proteins in fermented foods. Multiple peptides containing the R5 peptide QQPFP were present in LG beers at comparable levels to controls . B-, D-,The Finnish beer LG9 showed remarkably little evidence of gluten protein content. ALP peptide QQQQQGQSFVQPQQQVPVEITR and γ-hordein peptide APFVGVVSGVGGQ were present, as well as low levels of γ- and D-hordeins were detected. The lack of detectable B-hordein peptides suggests they were digested or removed to such a point that they did not cause ELISA signal suppression. The ELISA result of 2.0 mg/kg gluten  would thThese results highlight the difficulties faced when using ELISAs on fermented foods, and the practicality of testing the raw ingredients of fermented foods rather than the fermented or hydrolysed products. Hordein content measurements by R5 ELISA made after fermentation, in the presence of polyphenols, and estimated using certain standard materials have inherent inaccuracies. To ensure that gluten-free-accredited fermented foods contain <10 mg/kg gluten, measurements made using ELISA  must occur prior to fermentation, and the screening of the final product for contaminants must occur using a technique such as LC-MS, which does not encounter the signal suppression issues faced by ELISA. Nine LG beers that were specifically processed to remove gluten had less than 20 mg/kg gluten as measured by ELISA, though in all cases, specific hordein-derived peptides were detected using LC-MS, and in all but one, trace levels of wheat contamination were detected. LC-MS and ELISA measurements of hordein content revealed different relative abundances between the control and LG beers. All nine LG beers tested showed evidence of some hordein content, highlighting the importance of sensitive and reproducible sampling and gluten testing. Three LG beers contained peptides with intact epitopes defined as immunogenic for those with celiac disease, which is concerning because they may elicit symptoms in susceptible individuals if consumed at trace levels. Six LG beers had no immune reactive epitopes, one showed only trace levels of three hordein peptides, and eight of the nine had low levels of wheat gluten peptides, suggesting that contamination occurs at some point in production for most LG beers. This mismatch between ELISA and LC-MS is consistent with previous investigations that demonstrated ELISA-signal inhibition in certain beer types. Investigating the efficacy of gluten-reduction strategies (enzyme use or other proprietary methods) used in various breweries to lower gluten content would require further testing and optimisation. The variation in both total gluten content and presence/absence of immunogenic epitopes across beers that have been processed to remove gluten points to the challenges in gluten-free status assurance. Moreover, gluten presence due to ingredient or equipment contamination poses a further challenge for brewers. These factors are recognised in the recent FDA ruling on the gluten-free labelling of fermented foods. Given the challenges that remain in analysing partially hydrolysed (fermented) products, gluten quantitation of ingredients used in the production of gluten-free beers to test for the presence of contaminants is an appropriate approach, wherein sandwich ELISA is sufficient. Yet for beers produced using gluten-reduction strategies, wherein competitive ELISA may deliver variable results, techniques such as LC-MS offer an alternative for quality control assessment of GR beer production.


Background The role of temporal lobe epilepsy (TLE) in determining personality traits and neurobehavioral symptoms, collectively known as the interictal behavioral syndrome , as well as the syndrome's association with the particular artistic expression of many epileptic litterateurs are well known in neurology and psychiatry. A deepening of emotionality along with a serious, highly ethical, and spiritual behavior have been described as positive personality changes among patients with chronic mesial-TLE.Objectives Our narrative-based clinical hypothesis aims at contributing to the ongoing debate on the association between TLE and artistic expression, as well as the latter's supposed implication for epileptology in general and the neuropsychology of epilepsy in particular.Methods Through an analysis of the biography, language, and literary work of Greek novelist Demosthenes Voutyras, we hypothesize that his mystical and dark writing style could be attributed to medial temporal interictal dynamics.Conclusions We suggest that the psycholiterary profile of Voutyras is consistent with the idiosyncratic characteristics of the temporal lobe personality, while a non-dominant temporal lobe contribution has been proposed. Demosthenes Voutyras 1872–1958) is one of the most important Greek writers of the twentieth century, with an imposing work in terms of volume and quality.872–1958 He started writing as an escape from the devastating consequences of his illness, finding in it a way to express his desires and channel his great inherent vitality. Before devoting himself entirely to literature, Voutyras' father went bankrupt and committed suicide. Thereafter, his possessions were confiscated. Consequently, Voutyras was forced to get a job performing hard manual labor, which had a dramatic impact on him, making him get in touch with the life conditions and mind frame of the people who toil and struggle. The acquired social knowledge in combination with his innate talent transformed him in one of the greatest realistic storytellers in twentieth-century Greece .Voutyras was awarded the National Prize for Excellence in Letters and Arts 1924) and the Medal of the Municipality of Piraeus (1932). After the end of World War II, his honorary pension was terminated. Furthermore, the Academy of Athens, after two consecutive votes, rejected his application for membership, because Voutyras had supported the left-wing Greek Resistance during the Axis occupation of Greece in 1941–1944.4 and theHis stories have been translated and published in foreign anthologies and magazines. Many articles by foreign criticsMany of his works are also scattered in newspapers and magazines, in which his short stories began to appear in 1901. He published over 30 volumes from 1901 to 1945 .To our knowledge, Voutyras is the only known Greek fiction writer who officially suffered from early-onset epileptic seizures. His prose narrative works are almost exclusively linked to the social realism framework, and pioneering the transition, in Modern Greek literature, from ethography to urban prose.His world was that of the bourgeoisie and the urban working class. His imagery is reflective of nostalgic reminiscences of the poor quarters and suburbs of the city, and of the wretched life of its inhabitants.Voutyra's prose is characterized by precision in details blended with his perpetual imagination. leading to long descriptions (which are possibly hypergraphic due to his compulsively detailed accounts). This led to an anarchic style of writing rather than a structured work composition. Moreover, the psychological profiling of ordinary people seen in his writings, his dislike for social injustice and religious teachings, and, instead, his attraction for troubled atmospheres, set his technique apart as resistant to any esthetic analysis.The subject that dominated his writings was the life of marginalized people in Piraeus. Their life and mindset  were portrayed in melancholic colors, of almost utopic vision. These depictions take on an almost symbolic meaning,During the past decades, many clinicians undertook the challenging task of making retrospective diagnoses of nervous and mental diseases. Accordingly, in the second half of the twentieth century, talented neurologists gathered literary depictions of epileptic phenomena, such as those seen in the fictional characters created by Fyodor Dostoevsky (1821–1881) and Miguel de Cervantes 1547–1616). Dostoevsky's epileptic seizures have extensively been linked to his extraordinary literary style, and they represent the prototype of such pathographic inquiry. Along with other artists of the early twentieth century, Voutyras helped deepen neurobehavioral knowledge on epileptology and on the concept of temporal lobe personality. Using horror imagery, social drama, fiction, satiric reconstructions, and gothic style , he provided insight into the neuroesthetic dynamics of his epilepsy, probably TLE. This is consistent with previous studies assuming a right TLE diagnosis based on the works of writers, such as the eminent nineteenth century Brazilian litterateur Machado de Assis (1839–1908).–1616. DoDostoevsky's description of ecstatic auras received particular attention both from the general public and neurologists. Contradicting Gastaut 1978),,8who rIn the present essay, we describe the role seizures played in Voutyras' novels and the potential use of such information to retrospectively advance some diagnostic hypotheses regarding his condition through his literary style and neuroesthetic reflections. We occasionally provide instances of supposed seizure-related quotes. Finally, an effort was made to interpret these quotes in the light of the complex interplay between his writing style and seizure activity as determined by his prodromal and/or early ictal (the “aura”) manifestations.Chronic TLE causes mild interictal personality changes with a slowly but progressive cumulative effect.Hypergraphia, in particular, constitutes a symptom frequently observed in TLE. It is characterized by a strong desire toward compulsive writing, making detailed accounts of ordinary or personal life events or both, and writing poetry and fiction. It can even include lists or the mere repetition of written words, which are complex on a lexical and syntactic level. According to other authors,Involvement of the right cerebral hemisphere has been suggested, which is in line with right temporal interictal discharges in ecstatic auras.High religiosity scores on the Bear–Fedio Inventory and morphological abnormalities in the temporal lobe may account for the association between religious delusions and postictal psychosis;interictal personality disorderhas generated a great deal of controversy,The temporal lobe personality orRetrospective diagnosis has been criticized as an inappropriate method in medical history, and an interesting epistemic challenge against the retrospective diagnosis of historical figures points to the impossibility of scientific verification of a diagnosis in the past.For instance, syndromic diagnoses of functional disorders  and of many psychiatric disorders are made without any confirmatory test; yet, such diagnoses are highly practical accounts of the state of affairs of the patient in terms of treatment and prognosis. Of note, much of the criticism to such “anachronistic diagnoses” is decontextualized, since it often neglects the fact that different diseases might have coexisted in history, or that the same disease might have been described by means of different personal and clinical accounts, dictated by a particular historical context. Hapton e al.Some narratives by Voutyras resemble paradoxology and the horror inherent in the writings of H. G. Wells and Edgar Allan Poe,Voutyras's elliptical narratives and continuous time shifts from the past to the present and to the future are comparable to those of Joyce, Sartre and Faulkner. Temporal lobe epilepsy may induce aberrant time processing, in which the present may be mistaken for the past, and the past, for the present; the real may be seen as unreal, and vice versa; time may be sped up or slowed down.For Voutyras, time acquires a new role: it ceases to be the documentary time and becomes a parameter of consciousness. There is a gradual contraction of time from one pair to another  which intensifies the myth and the events in it. Narrative fluidity is also remarkable. Indeed, both narrative space and time constantly change, in an attempt to highlight the internal elements of the myth. At the same time, the writer is somewhat “indifferent” to their external organization..Voutyras's style breaks the rules of ordinary spontaneity and structure of writing. Without predefined rules, he deliberately combines fantasy and reality,. He forms a purely personal narrative time, “playing” with what the reader's expectations with constant twists.As Dimarasseizures and epilepsy used as metaphorsorseizures described in metaphors.The relationship between epilepsy and literary metaphors, may be expressed by means of eitherIn fiction, both the above are likely to reflect societal stereotypes, while seizures are conceived as bordering on religion and sexuality, as expressions of critical emotions ending in ecstasy or breakdown. Approximations to the experience of a seizure typically refer to either dreams or intensive sensory experiences. Of note, the use of metaphors by writers suffering epilepsy and by those who possess knowledge on epilepsy differs only superficially.Epileptic auras may represent a precious instrument in the way many litterateurs who are sufferers perceive their own selves and the world. In addition, the study of these experiential phenomena may promote understanding of new and enlightening aspects of epilepsy. The bizarreness and unusualness of aural and ictal subjective experiences are of such an “intensity” that patients may lack the expressive means to describe them. This is probably the reason why self-reports of epilepsy often are rich in metaphors and oxymora.For instance, in his narrative “The city of the curse”, the following oxymoron can be seen: “and the child with the old face who came to be paid and he smelled like that [...]. But he was not a child, he was an old man with something childish!”Superstitions and feelings of prescience expressed by his characters – often encountered in seizures of temporal lobe origin – are key elements in his stories. In “The revenge that will come”, one reads: “one morning he got up with a premonition that something bad was going to happen. He had also seen a dream”.The oneiric atmosphere of his prose makes the reader discover dream worlds through suggestion techniques. It is believed that Voutyras possessed a deep knowledge of the troubled mind, probably because of his own experience with epilepsy.These stories abound with cemeteries, blood rites, murders, scary dreams, dim lights, premonitions, obsessions, and illusions, all impregnated with a deadly savagery, and possibly rooted in the artist's temporo-limbic epileptogenesis. Voutyras portrayed heroes surrender after fighting in vain the darkness: “With the one who secretly directs these blows, how was it possible for him to fight? This is hidden everywhere, in the light of day, in the dark, in the air!”.“The darkness, or its shadow, began to fall, and in it the black companions of the night began to fly fast […].”“at that moment, along with it, a shadow, a dark figure of a man stood on the side of the road […]”“the wind did not blow, and the giant heavy-headed pines, the tall cypresses remained motionless, black, dark […]”“I was on a dark and mournful street in Piraeus […]”“and the night came wild, dark as hell, and like the sword of an angel the lightning tore it every now and then […]”“someone in the dark with an ax dug a tree trunk to make a baby crib […]”Interestingly, indications of weakness and vulnerability are often met as prototypes of epilepsy metaphors.As Voutyras stated,The first blow came to me in lower secondary school. I fell down during the lesson. Others tried in vain to help me recover. Where I was thinking something and somewhere my imagination was dragging me […] something was presented and with that I fell. This thing was presented as an old, forgotten memory. And it was so sweet that it led me for a while to a state of non-existence […].In “The revenge that will come” a déjà vu experience becomes evident:Paul listened carefully, but suddenly something circled him like a dream, or a piece taken from sleep, and it seemed to him then that what the teacher had said, the piece of Scripture and that he would still say, if he knew everything, everything, someone to have told him, to have read them to him on a card at other times, in mysterious hours!Voutyras began his writing carrier at the age of 35 years. Τhat illness that had oppressed me so much, had stopped bothering me […]. It did not bother me anymore from the day I lost my way to music […]. After he did everything, so I can say, to throw me in the literature, he left me”. He studied to become an officer in the Greek Merchant Marine Academy, a profession that he never pursued because of his seizures; moreover, he practiced fencing, and later on took tenor lessons. Epilepsy, however, distanced him from a music and a career as a tenor. And, as he himself wrote, “like a hand or a kick, it pushed me and threw me into literature”.This piece of autobiographic evidence leads us to hypothesize that the Greek writer suffered déjà vus linked to seizure activity of temporal lobe origin. Such a highly relevant clinical information, along with the general picture gathered through the idiosyncratic qualities emerging from his work, further support our literature-based hypothesis. In “The Prescience”,“But I will tell you now, how wild animals, birds, vultures, cry, mourn when it is to fall in the place where they have lived for years, in their homeland, some great calamity! […] I rushed out, on the street. But as soon as I came out from the shadow of the house and was illuminated by the light of the moon, a different voice, wild with fear, was heard coming from nearby […]I raised the wood. I did not see anything. “But what is this!”, I said. Isn't it a night owl, owl? […] Isn1t it the owl of Athens, and it will be, and it cried for some great evil, great evils, that will fall in Athens and Greece?Mystical experiences are widely linked to temporal lobe function,In “The revenge that will come”, an ecstatic aura can be observed:Τhe teacher  was talking, talking, but he was no longer paying attention, he was immersed in that magician, who, like a cloud invisible to others, had surrounded him and made him feel so much, and as if he heard other voices saying to him.In conclusion, we suggest that the psycholiterary profile of Voutyras is consistent with the idiosyncratic characteristics of the temporal lobe personality, while a non-dominant temporal lobe contribution has been proposed. One might indirectly discern a sort of resemblance of Voutyras epilepsy, abounding of ecstatic auras and déjà vus, mystical and dark elements and profound analytic detailing, with the aberrant interictal temporo-limbic dynamics  encountered among renowned artists suffering from TLE and presenting with temporal lobe personality. The lack of composition, put forward as a criticism to his work, might be linked to the fundamental disruption of self-awareness induced by seizures. Moreover, several writers suffering from epilepsy have used aura experiences in their works,Finally, from a conventional scientific viewpoint, Voutyras's epilepsy type, like in the case of Dostoevsky, may easily turn “into a 'cul-de-sac' debate”Last but not least, detecting qualitative information on retrospective diagnoses from the literature may represent an indirect way of reviving the clinical acumen that once was of vital importance to clinicians. Those professionals were able to formulate diagnostic hypotheses and subordinate data to their clinical reasoning, “not the inverse, as passive pursuing of instrumental data makes clinical reasoning follow instrumental data as a slave follows his master”, as Luria advised.


Fruit from treated and untreated vines was harvested 45 days after CaCO3 spraying and evaluated for quality. The fruit was vinified, and the obtained wines were bottled and stored in darkness for 15 months at 20 °C. Wines were evaluated for quality after storage. The evaluation of grape and wine quality included the content of phenolic compounds and antioxidant capacity. The treatment with CaCO3 did not affect the ripening rate of grapes. However, the treatment improved the fruit yield as well as the color development, the content of phenolic compounds, and antioxidant capacity of grapes and wine. The treatment favored especially the accumulation of malvidin-3-O-glucoside, pelargonidin-3-O-glucoside, caftaric acid, caffeic acid, trans-cinnamic acid, quercetin, catechin, epicatechin, resveratrol, and the procyanidins B1 and B2. Wine made with treated fruit was of higher quality than that of control fruit.The quality of wine grapes and wine depends on their content of phenolic compounds. Under commercial conditions, the phenolic maturity of grapes is mostly achieved by applying abscisic acid analogues. Some Ca forms represent a cost-effective alternative for these compounds. In this study, ‘Shiraz’ vines (veraison of 90%) were sprayed with CaCO The content and diversity of phenolic compounds determine important quality attributes of red wines, including color, astringency, bitterness, and beneficial effects on human health, among others ,2. Under2 increased the accumulation of anthocyanins [3 increased the concentration of anthocyanins, flavan-3-ols, and other groups of phenolic compounds in ‘Shiraz’ grapes during ripening. The preharvest treatment with Ca also induces alterations in the mechanical properties (hardness and flexibility) of grape skin [3 on the biosynthesis of phenolic compounds in ‘Shiraz’ grapes and evaluate the content of phenolic compounds in wine made with these grapes after short-term bottle aging.The preharvest application of Ca seems to be an economically feasible alternative to improve the phenolic content in grapes and other fruits. Some studies have demonstrated that the foliar application of Ca increased the expression of phenylalanine ammonia lyase (PAL) and the phenolic content in grapes . Ca alsoocyanins ,12,13. Rocyanins  also demape skin . This adape skin . The comape skin . CurrentVitis vinifera) grafted on the 1103 Paulsen rootstock. The plantation is oriented from north to south with a spacing of 3.0 m between rows and 1.0 m between plants with a Royat system in sandy loam soil. The simple drip irrigation system was applied from sprouting to the beginning of veraison, maintaining the average soil moisture around 16% during these vegetative stages. Irrigation was restricted during ripening, from veraison until harvest time, however, the rains were frequent during ripening in the site of study . The vineyard was established in 2009 with the ’Shiraz’ variety , SiO2 (33.77%), free Ca (24.4%), and free magnesium (0.32%). The application of the residue solution was performed using a backpack-type automatic sprinkler. Unsprayed plants were used as the control group.Residues from a local cement industry were used as the Ca source at a concentration of 1% in water, using Tween-20 (0.05%) as the dispersant agent. This concentration of residues was determined based on preliminary studies with concentrations of 1 and 2%. The total content of Ca in this solution was 4.26 g of Ca per L. The treatment of vines with this solution was referred in this study as the treatment with CaCO3 solution. Bunch weight was determined at harvest time. The individual weight of grapes and their proportion of skin, seeds, and pulp were determined in 50 fruits per treatment and replication. Skin and pulp samples were oven-dried at 100 °C until constant weight and then ashed in a muffle at 550 °C for 8 h. The ashes were treated according to Fernández-Hernández et al. [L*, a*, b, C*, and °h) was determined using a CR-300 colorimeter . Fruit firmness was determined with a TA-XT2i texturometer  equipped with a 7 mm diameter stainless steel striker pin, which punctured the fruit at a rate of 10 mm s−1. The maximum puncturing force was recorded in N. These fruits were pureed in a kitchen blender immediately after firmness evaluation and the puree was evaluated for moisture content, the content of total phenolics (TPC), individual phenols, and antioxidant capacity (AC). The juice obtained from the puree was evaluated for °Brix, pH, and titratable acidity (TA). The moisture content of fruit was determined gravimetrically in puree samples (3 ± 0.5 g) dried at 100 °C until constant weight. The °Brix, pH, and TA were evaluated using a digital refractometer  and pH meter . TA was determined by titration at a pH of 7.0 with 0.1 N NaOH.Fruit from treated and untreated plants was harvested 45 days after the application of the CaCOz et al.  and eval−1 of SO2 and 24 h later, inoculated with enological Saccharomyces cerevisiae  at a proportion of 0.2 g L−1 in individual tanks of 40 L. During the alcoholic fermentation, the temperature and the decrease in °Brix were monitored until the completion of the alcoholic fermentation. The wines were pressed, and the malolactic fermentation was carried out at a controlled temperature of 20 °C. The wines were clarified with Vinigel Platinum at a rate of 0.05 g L−1 , subjected to tartaric stabilization at 2.0 ± 1 °C, placed into 750 mL amber bottles, which were closed with natural corks  and a heat-sealable capsule. The bottled wines were aged in complete darkness for 15 months at 20 °C. The wines were analyzed for alcohol content by densimetry, pH after elimination of carbon dioxide, TA by titration with 0.1 N NaOH, and chromatic characteristics of intensity and tonality at 420, 520, and 620 nm using a GENESYS 10 S UV-Vis spectrophotometer . Additionally, the density of wines was determined using a pycnometer . The evaluations were performed according to the international methods of the International Organization of Vine and Wine [Another lot of grapes was divided into four groups per treatment. Each group was composed of 30 kg of destemmed grapes. These grapes were vinified at 20–23 °C under red light. The musts were sulphited with 50 mg Kgand Wine . Wines wg, 5 min, 25 °C). Then, the supernatant was recovered and diluted with deionized water . The fruit and wine extracts were filtered through a nylon membrane of 0.45 μm pore size and analyzed for total and individual phenolic compounds. TPC was determined by Folin Ciocalteu’s method at 750 nm using a Bio-Rad microplate reader model 680 . The quantification was performed using gallic acid as the reference compound and the results were expressed as mg of gallic acid equivalents (mg GAE) per Kg of grapes, on a fresh weight basis, and as mg GAE per L of wine.The phenolic compounds were extracted from grape puree according to previous studies . For win2 = 0.973 to 0.999) constructed with three independent sets of dilutions of standard compounds. The instrument LOD and LOQ for phenolic compounds ranged from 0.000016 to 0.0440138 and from 0.0000743 to 0.1467126 mg mL−1, respectively.For the evaluation of individual phenols, the extracts were injected (20 μL) into an Agilent 1200 HPLC system equipped with a diode array detector . The separation of phenols was carried out at 30 °C in a Zorbax XDB-C18 column (4.60 × 150 mm)  using the gradient of solvents reported previously in other studies . The ind−1). The samples were incubated for 30 min in dark conditions until measurement at 515 nm . The % of inhibition of DPPH was recorded.The extracts used for the analysis of phenolic compounds were also evaluated for AC according to Abbasi et al. . Brieflyp < 0.05 as the significance limit. SAS software Ver. 9.0  was used for data analysis.The experiment was carried out in quadruplicate using a completely randomized design. The data were analyzed using ANOVA followed by the Tukey–Kramer post hoc test with 3 increased significantly the weights of clusters (82%) and berries (46%) (The treatment with CaCOes (46%) . The mages (46%) . Ca did 3 treatment  and higher pH (6%) as well as greater intensity of color (41%) and tonality (6%) compared with wines made with grapes from untreated vines , flavan-3-ols , anthocyanins , and the stilbene trans-resveratrol. It caused increases of several times in the content of most of these compounds in grapes  treated in preharvest with CaCl2, causing increases in weight of bunches and individual berries [2 also increased the yield in pomegranates [PG1 and PG2 gene expression, the promotion of sugar accumulation in fruits, and the Ca-mediated signaling that allows the expression of VIT_02s0012g02190, VIT_16s0039g02020, and VIT_18s0072g00370 genes, which cause increases of cellulose content in fruit [VIT_03s0038g03430 and VIT_08s0007g00440, responsible for cell expansion and fruit growth, or the FaCDPKs, gen of CDPKs involved in fruit development, as reported in grapes and other berries [The effects of Ca on plant nutrition and yield and firmness of many fruits have been widely studied ,22. Howe berries ,21,22,23in fruit ,15. Addi berries ,13. Ca c berries ,24,25,263 did not affect the ripening rate of grapes, according to their values of TSS, pH, and TA. Similarly, the foliar application of CaCl2 and ABA on pomegranate plants and vines, respectively, affected some quality attributes of fruit but SST, pH, and TA remained unaltered in fruit [3. This effect might be related to other compositional changes in fruit induced by the treatment with CaCO3, with these changes altering the performance of fermentative microorganisms and enzymes during fermentation and, consequently, retarding the stabilization of the content of organic acids  in the must, as observed in ’Muscadine’ wines [The treatment of vines with CaCOin fruit ,22,27. He’ wines .3 also increased the firmness of grapes, as reported for other grape varieties  cultivated in other regions and treated in preharvest with CaCl2 [2 [3-treated grapes could also influence the release of grape juice and phenolic compounds during the wine making process, causing higher content of phenolic compounds in wine. Similarly, the treatment of pears with CaCl2 caused increases in fruit firmness and release of juice [The treatment with CaCOth CaCl2 ,21. Thisth CaCl2 . AlthougCaCl2 [2 . This inof juice .3-treated vines showed a darker color, characterized by lower L* and b* values than those of fruit from untreated vines. Similarly, others have observed that the preharvest treatment of vines with ABA favored color development in ‘Crimson Seedless’ table grapes and fruit from some wine grape varieties  [3 on the biosynthesis of phenolic compounds might be related to its effect as a stress factor and direct interaction with enzymes involved in the phenylpropanoid pathway, improving the color of grapes and wine [Grapes from CaCOMerlot’) ,8,31,32.Merlot’) . As expeMerlot’) . MeJA acMerlot’) ,34. Thusand wine .3 caused increases in the contents of total and individual phenolic compounds in grapes. The effect of Ca on these response variables has been previously reported for some fruits, including ‘Sweetheart’ and ‘Lapins’ cherries and strawberries treated in preharvest with CaCl2 [3 on the content of total and individual phenolic compounds in aged wine. Our study demonstrated that the treatment of vines with CaCO3 improved the content of these compounds in wine. This finding is supported by studies with ‘Yan73′, ‘Merlot’, and ‘Cabernet Sauvignon’ wines made with fruit from vines treated with ABA in preharvest, which showed a higher TPC than control wines [On the other hand, the treatment with CaCOth CaCl2 ,35. ‘Malth CaCl2 . Currentol wines ,32.3-treated vines and their wine showed higher contents of malvidin-3-O-glucoside and pelargonidin-3-O-glucoside. Similar results have been observed in table grapes and strawberries from plants treated with CaCl2 in preharvest and cherries treated with CaCl2 in postharvest [3-treated grapes contained pelargonidin 3-O-glucoside, an anthocyanin that has been reported in ‘Cabernet Sauvignon’ and ‘Pinot Noir’ grapes from vineyards established at an altitude above 2000 m.a.s.l. [3 on malvidin-3-O-glucoside was lower in wine than in grapes, probably due to CaCO3 inducing the synthesis of other polyphenols not identified that copigmented with this anthocyanin, decreasing the concentration of its free form. As stated above, there is no information about the effect of the preharvest application of Ca on the composition of phenolic compounds in aged wine. In our study, the treatment of grapes with CaCO3 caused increases in the anthocyanin content in wine. This Ca-mediated effect was similar to that reported for ‘Yan73’, ‘Cabernet Sauvignon’, ‘Malbec’, ‘Tempranillo’, and ‘Graciano’ wines made with grapes from vines treated with ABA or MeJA in preharvest [3 on the anthocyanin content in grapes and, consequently, in wines. Ca activates directly the biosynthesis of ethylene, ABA, and other phytohormones, which influence the biosynthesis of anthocyanins [VIT_08s0040g00470 and VIT_14s0006g01400) and CDPKs (VIT_04s0023g03420), causing increases in the expression of some genes involved in the synthesis of phenylalanine ammonia-lyase , 4-coumarate CoA ligase , UDP-glucose: flavonoid 3-O-glucosyltransferase , which play an important role on anthocyanin biosynthesis in grape skin [O-glucosyltransferase, dihydroflavonol 4 reductase and anthocyanin synthase, as reported in some fruits [MdMYB308L, MdbHLH33, MdDFR, etc.) involved in anthocyanin biosynthesis, as reported in apples [3 also favored the biosynthesis of trans-cinnamic acid in grapes, which is a precursor of anthocyanins [3-mediated increase of skin mass in grapes, with skin being the major accumulation site for anthocyanins [In our study, grapes from CaCOtharvest ,34,35. Atharvest ,35. Intem.a.s.l. . The poseharvest ,32,33. Socyanins . Howeverase PAL,  genes, 4ase PAL,  genes, 4e fruits ,38. Finan apples ,39,40,41ocyanins .3 favored the accumulation of some flavonols in wine, but they were not observed in grapes. Gil-Muñoz et al. [3-treated grapes. Others have also observed that the treatment of vines  with MeJA or ABA caused increases of quercetin in wines [3 favored the accumulation of quercetin in the tested samples since the accumulation of this flavonol is mainly influenced by exposure of fruit to light, especially to UV-B light [The treatment of vines with CaCOz et al.  did not z et al. . Quercetin wines ,33. Thisin wines . However-B light ,43.3 caused increases of flavan-3-ols in grapes and wines, including catechin, epicatechin, and the procyanidins B1 and B2. Similar increases of flavan-3-ols have been observed for ‘Yan 73’, ‘Cabernet Sauvignon’, ‘Merlot’, and ‘Monastrell’ grapes and wines from ABA- or MeJA-treated vines [The treatment with CaCOed vines ,42. The ed vines . These ced vines .3-treated vines showed a higher content of the stilbene resveratrol compared to grapes from untreated vines and their wine. The preharvest treatment of ‘Malbec’ vines with ABA and MeJA also caused increases of stilbene in fruit and wine [3 and CaCl2 also increased the stilbenoid content in ‘Shiraz’ and ‘Vinhão’ grapes, allowing the inference that Ca and MeJA act similarly on stilbene biosynthesis [2 [Grapes and wine from CaCOand wine ,33. Howeand wine ,33. Inteynthesis ,15. The ynthesis ,13,24. Synthesis  and in ‘hesis [2 .3 favored the biosynthesis of trans-cinnamic and caftaric acid in grapes. In contrast, Portu et al. [trans-cinnamic acid found in grapes from CaCO3-treated vines is beneficial from the enological point of view because some studies have demonstrated that this acid is the precursor of other phenolic compounds, including its derivatives [trans-cinnamic acid favors the copigmentation of phenolic compounds with flavan-3-ols and the color stabilization of wine during aging [3-treated vines because it prevents oxidation of wine during aging, favoring its stabilization [The treatment with CaCOu et al.  did not u et al.  did not ivatives . This inng aging . These blization .3-treated vines could be attributed to the cascade of Ca-mediated signaling reactions to activate the expression of CaMs, CDPKs, and CLBs proteins, and other mechanisms indicated above. These proteins participate as sensors and transmitters in response to biotic or abiotic stress in plants, as seen in many plants under salinity or drought conditions [3 seems to act as a secondary messenger involved in the adaptability responses of vines to biotic and abiotic stress, promoting the synthesis of ABA or JA and, consequently, of phenolic compounds [3 could also induce the synthesis of other enzymes involved in the biosynthetic pathway of phenolic compounds [3-mediated increase of skin mass in grapes could also be involved in the increases of phenolic compounds [2-treated vines [2 and ‘Cabernet Sauvignon’ and ‘Merlot’ grapes with ABA caused increases of AC in fruit and wines, respectively [3, as other elicitors , is able to induce the accumulation of phenolic compounds in grapes and wine, causing increases in AC and, probably, in the activity of these foods in the prevention of some chronic degenerative diseases [Overall, the increased content of total and individual phenolic compounds in fruit and wine from CaCOnditions . This stnditions ,11. Thusompounds ,13. CaCOompounds . These ied vines . The preectively ,12. Our 3 on ‘Shiraz’ vines improved the phenolic composition, antioxidant capacity, and other quality attributes in grapes and wine . CaCO3 certainly acted as a biostimulant and its effects were similar to those reported for fruit and wines from vines treated with other biostimulants, most of them expensive. Thus, CaCO3 represents a cost-effective alternative to improve the quality of grapes and wine.The foliar application of CaCO3-treated vines is an important source of phenolic compounds and other antioxidant compounds, allowing the inference that the consumption of this food in moderation might have an increased beneficial effect on human health, especially preventing several degenerative diseases. Wines are considered an excellent source of compounds with cardio protective effects.Wine from CaCO