Norovirus (NoV) and hepatitis A virus (HAV) pose a considerable health risk worldwide. In recent years, many cases of virus infection caused by virus‐contaminated strawberries have occurred worldwide. This study applied a critical control point system to analyze the main hazards during the production and marketing of strawberries imported into China and explore the key control points in the whole process. To further evaluate the risks in the supply chain, the established quantitative microbial risk assessment (QMRA) was used to determine the probability that residents would be infected with viruses after consuming imported strawberries. It was found that the risk of virus contamination from imported strawberries was low, and the virus contamination mainly results from water resources and personnel. This research helps the regulatory authorities formulate strategies to ensure the long‐term microbial safety of imported strawberries. In addition, the methods may prove useful in evaluating the risks of other agricultural produce. Enteroviruses are the main cause of outbreaks of foodborne diseases. We investigated the risk of norovirus and hepatitis A virus in imported strawberries based on the supply chain from both qualitative and quantitative perspectives. In the strawberry supply chain, the virus contamination factors were determined by the critical control point system. The QMRA allows uncertain factors to be incorporated and propagated into the model using input values following a certain frequency or probability distribution function and the Monte Carlo simulation  stipulates that the preharvest interval of crops should not exceed 14 days in most cases and be at least 7 days. The United States Environmental Protection Agency (https://www.epa.gov) stipulates that the preharvest interval of strawberries is generally 14 days. Given the similarity of the collected data between the two countries, the two countries were deemed to have adopted the same safety interval in this study. The interval range for different pesticides was set to 0–14 days, considering any possible illegal operations. During the picking process, the virus from the strawberries is transferred to the hands of the workers, and the virus from the hands of the workers is transferred to the strawberries, so the contamination is reciprocal. Picking may cause cross‐contamination of NoVs between the agricultural products and the workers' hands , and Nharv is the number of viruses on the hands of workers .Many pesticides have a defined preharvest interval which is the time between the last pesticide application and the harvest. During the preharvest interval, the virus will be exposed to the environment and loses viability over time. The Australian Pesticide and Veterinary Drug Administration  among the infected population was used. The values of η and 2.2.7The extracted data were entered into a spreadsheet Table , and eac33.13.1.1According to the strawberry supply chain Figure , the expMost virus outbreaks related to fresh fruits are caused by irrigation with polluted water in the field  < PSAustralia (−2.92 LogPDU/berry) < USAmerica (−1.28 LogPDU/berry) < USAustralia (−1.22 LogPDU/berry) < FSAmerica (0.0166 LogPDU/berry) < FSAustralia (0.0168 LogPDU/berry). Compared with the same kind of strawberries, the pollution level of NoV in Australian strawberries is higher than that in American strawberries. This result is consistent with the information published in the two national institutions. The average annual number of NoV infections reported by the Center for Disease Control and Prevention (https://www.cdc.gov/) in the United States is 601,378. The Australian Ministry of Health  reported that, on average, 1,800,000 Australians are infected with NoVs every year.As shown in Figure https://www.cdc.gov/) excluding the homeless is 3025. The annual average number of hepatitis A cases in Australia  is 299. The population density of the United States and Australia (httpsww.worlddata.info/) is 33 and 3, respectively. The severity of the food‐related hepatitis A outbreak was expressed by the ratio between the average number of hepatitis A cases to the population density, which was 90.6 in America and 91.2 in Australia. Therefore, the outbreak degree of food‐related hepatitis A is consistent with the pollution results calculated in this study.The statistics shows that the main reasons for the outbreak of hepatitis A are the health problems of the homeless and the virus contamination of food. There are no outbreak cases of homeless groups in Australia. The average annual number of hepatitis A cases in the United States  into the main operating system interface  architecture, this section adopts Java EE and MVC (Model–View–Controller) models to design and implement a pathogen quantitative risk assessment system. The Model is used to store and process data, the View is used in the application to realize the visualization of risk data, and the Controller is used in the application to deal with the interaction of risk analysis users. The MVC pattern is used to identify the client and server components in the Web‐based online system, and the user interaction information and interface are modified and formulated. The platform realizes the information of virus damage in the production and marketing chain of strawberry, can report the potential risk of pathogen contamination in strawberry early, improve the efficiency of risk assessment, and has certain application value. Users can through the website links  and the Centers for Disease Control and Prevention (www.cdc.gov/infectioncontrol/guidelines/norovirus). Government departments should conduct management and sampling inspections on their strawberry markets. The results of strawberry detection should meet the requirements of DBS13/001‐2015  to query the trade of strawberries, we can find that the United States and Australia are closer to China's strawberry trade and other countries are not necessarily in our main trade objects. In less developed countries, there may be more people infected with NoV, so quantitative risk assessment models can be built for more less developed countries in the future to provide a virus contamination warning platform. Strawberry production and health management of workers need water resources, so it is important to understand the fate of viruses in water and wastewater.Combining the critical control point system with the QMRA model, the study can effectively determine the virus contamination factors in strawberry production and marketing, ensure the quality and safety of strawberries, and provide key quality control strategy references for managers. Our study shows that NoV and hepatitis A in many berries originate from water and food handlers, which are supported by some investigations in Europe, North America, Asia, Australia, Africa, and South America  study u5In this study, we investigated contamination risk of NoV and HAV in imported strawberries through qualitative and quantitative analyses across the supply chain. Employing a critical control point system, we identified field management and disinfection as pivotal in minimizing virus risks. Our research adhered to rigorous hygiene standards while contributing novel insights. Based on the critical control point system, the QMRA model was explored which revealed distinct contamination levels based on strawberry origin, aligning with authoritative agricultural data which showed that the contamination levels of virus in strawberries from Australia were higher than those from the United States, consistent with the information displayed by the agriculture departments of the two countries. We highlighted the disparity in safety between carefully processed and directly picked fresh strawberries, emphasizing the importance of postharvest measures. Notably, our study unveiled heightened virus persistence on FS, underlining the need for tailored handling during freezing. Our pioneering risk assessment early warning platform equips researchers and authorities with predictive tools for timely interventions.Overall, our research advances food safety understanding, offering practical risk management strategies while enhancing the safety and quality of strawberries across the supply chain.Junjie Zhong: Writing – original draft . Yunfeng Yang: Visualization . Hui Zhang: Validation . Shuwen Zhang: Validation . Xiaosheng Qu: Validation . Qin Chen: Supervision . Bing Niu: Project administration ; writing – review and editing .This study was supported by the Shanghai Agriculture Project (19391901500) and the National Key Research and Development Program of China .The authors declare that they have no conflicts of interest.


Various viral infections can affect the oral cavities of pediatric patients ,2. Some Viral infections of the oral cavity can be distinguished into those that do not result in visible damage or a disease in the oral cavity, but are transmitted orally or during dental procedures, and those associated with oral and perioral lesions ,3. HowevPrimary infection with herpes simplex virus-1 is generally acquired during early childhood and is usually subclinical or underdiagnosed . The priHuman papillomaviruses (HPV) can cause benign and malignant diseases in various areas, such as the genital and oral mucosa or skin . InfectiSARS-CoV-2 infections occurred less frequently among children than they do among adults during the early phase of the pandemic, possibly because infected children were asymptomatic or had only mild symptoms . HoweverEnteroviruses are RNA viruses that are usually fecally–orally transmitted . They beMorbilli virus is a negative-stranded RNA virus that belongs to the paramyxovirus family. It is transmitted via direct contact and has an incubation period of less than one week . MorbillMumps virus infection typically leads to inflammation and the swelling of the parotid gland, resulting in parotitis ,24. The Human immunodeficiency virus type 1 is most commonly transmitted vertically in children from an infected mother during childbirth, in utero, or through breastfeeding ; althougVaccination programs have effectively prevented severe childhood infections, although viral infections remain a serious health problem in developing countries ,25. Emer


To examine direct and indirect costs, early retirement, cardiovascular events and mortality over 5 years in people with atherosclerotic cardiovascular disease (ASCVD) and matched controls in Sweden.Individuals aged ≥ 16 years living in Sweden on 01 January 2012 were identified in an existing database. Individuals with ASCVD were propensity score matched to controls without ASCVD by age, sex and educational status. We compared direct healthcare costs , indirect costs (resulting from work absence) and the risk of stroke, myocardial infarction (MI) and early retirement.After matching, there were 231,417 individuals in each cohort. Total mean per-person annual costs were over 2.5 times higher in the ASCVD group versus the controls (€6923 vs €2699). Indirect costs contributed to 60% and 67% of annual costs in the ASCVD and control groups, respectively. Inpatient costs accounted for ≥ 70% of direct healthcare costs. Cumulative total costs over the 5-year period were €32,011 in the ASCVD group and €12,931 in the controls. People with ASCVD were 3 times more likely to enter early retirement than controls (hazard ratio [HR] 3.02 [95% CI 2.76–3.31]) and approximately 2 times more likely to experience stroke (HR 1.83 [1.77–1.89]) or MI (HR 2.27 [2.20–2.34]).ASCVD is associated with both economic and clinical impacts. People with ASCVD incurred considerably higher costs than matched controls, with indirect costs resulting from work absence and inpatient admissions being major cost drivers, and were also more likely to experience additional ASCVD events.The online version contains supplementary material available at 10.1186/s12872-023-03518-y. Cardiovascular disease (CVD) affected over 120 million people globally in 2016 , which iBoth CVD , 7 and AA small number of cost-of-illness studies have shown that ASCVD is associated with indirect costs due to lost productivity , 14–17; Population-based administrative registries hold large amounts of individual-level, real-world data on the occurrence of disease, healthcare resource use and work absence. In addition, using population-based registries often means that multiple data sources can be connected to allow exploration of the broader economic burden of common conditions. For example, linkage of healthcare resource use to subsequent mortality or work absence for people with defined diagnoses permits evaluation of outcomes over time and helps researchers to understand the wider societal impact of disease and interventions , 18–20. In the present study, we examined direct healthcare costs, indirect costs resulting from work absence, early retirement, stroke, myocardial infarction (MI) and mortality over 5 years, comparing individuals with ASCVD and matched controls without ASCVD. In a separate analysis, we examined the impact of the first occurrence of ASCVD on direct and indirect costs.This observational, retrospective, closed-cohort study used an existing database at the Swedish Institute for Health Economics, originally set up to identify individuals with diabetes, plus control individuals without diabetes, between the ages of 16 and 70 years. Details of patient and control selection have been published previously , 20. In n = 491,116) plus controls  in the existing database. ASCVD was defined as the presence of at least one inpatient admission or outpatient visit with a main or sub-diagnosis of cerebrovascular disease, ischemic heart disease, or PAD  and activity compensation (long term absence/early retirement). Indirect costs and early retirement were analysed for individuals aged under 66 years, i.e. labour-market active ages.In addition to direct and indirect costs, the risk of four types of events were analysed and compared between the ASCVD and control groups: full-time early retirement, an inpatient admission due to stroke  or MI (ICD-10 code I21), or death at any time during the study period. To ensure that the ASCVD-related diagnosis was the main cause of hospital admission, the analysis of events used a stricter definition to that used in the main analysis and required that stroke or MI was the main diagnosis for inpatient admission.Annual total, direct and indirect costs were assessed in a subset of individuals with their first observed occurrence of ASCVD in 2011 (the year before baseline). Mean annual per-person costs for the years 2007–2016 were compared with costs for controls to quantify the impact of incident ASCVD.Mean and standard deviation (SD) are reported for continuous variables, and categorical variables are reported as actual numbers and proportions (%). Standardized differences for baseline characteristics were calculated after matching using the Stata package ‘stddiff’, and a standardized difference of greater than 0.1 was considered to indicate that groups were not balanced. Cost estimates are reported as mean and standard error (SE). Fine–Gray competing risk regression analyses  were used to estimate the hazard ratios (HRs) and 95% confidence interval (CIs) for full-time early retirement, stroke, MI and death, with adjustments for non-CV conditions . Individuals who were already in early retirement before baseline were excluded from the estimation of the HR for full-time early retirement.Potential violations of proportionality assumptions were tested using visual inspection of log–log plots and observed Kaplan–Meier plots against Cox predicted curves. Schoenfeld residuals were also calculated for each regression variable, with the null hypothesis of proportional hazards.We identified 231,417 individuals with ASCVD in at least one related observation before the study period (01 January 2012). From a pool of 2,088,675 potential controls without ASCVD, 231,417 individuals were matched 1:1 based on age, sex and educational status. The numbers of individuals in the control group who subsequently met the criteria for ASCVD in each year following the start of the study are shown in Supplementary Table SStandardized differences of the matching variables indicate similarities between individuals with ASCVD and the controls Table . In bothMean per-person annual costs were more than 2.5 times higher in the ASCVD group  than in the control group , with a similar pattern for cumulative total per-person costs over 5 years . The greatest contributor to these totals was indirect costs, which contributed 60% of annual and 60% of cumulative costs in the ASCVD group  and 67% of annual and cumulative costs in the control group  compared with controls (52 days). For those with ASCVD, on average, 8% were absent for 180–269 days per year and 17% were fully absent each year. For controls, 4% were absent for 180–269 days and 7% for more than 360 days per year. Among individuals with ASCVD, 52% had no work absence in the year 2012, compared with 75% of controls.n = 21,351 in 2012), annual costs were examined from 2007 to 2016 and compared with controls. Total costs decreased gradually decreased from 2007 to 2010 in individuals with observed ASCVD but increased sharply in 2011, from €6263 in 2010 to €18,538 in 2011. This increase in the ASCVD group was mostly driven by direct costs, which increased from €920 in 2010 to €12,188 in 2011 . The trend for a decrease in indirect costs over time is likely to be the result of population aging, whereby the proportion of individuals reaching retirement age counteracts the impact of work absence on costs.n = 38,352) of individuals in the ASCVD cohort died, compared with 8.7%  of controls, and all-cause mortality was 1.7 times higher (HR 1.7 [95% CI 1.7–1.8]) in those with ASCVD than in controls  or MI  than their controls  by including individuals who had at least one registered occurrence of ASCVD at a hospital. The primary prevention population  may also benefit from preventative interventions and should be evaluated in future studies. A limitation of this study concerns the fact that the existing database only included individuals with diabetes onset before the age of 70 years and their controls, and that although follow-up spanned up to 20 years, the dataset is likely to be under-representative of the very elderly section of the general population. Data on primary care were not available for this analysis and, although hospital costs are expected to be a comparatively far larger contributor to the overall economic burden of CVD, this means that the total costs in this study are likely to be underestimated. The contribution of primary care to healthcare costs is also likely to vary between different manifestations of ASCVD; further investigation of this is warranted.This study demonstrates that ASCVD incurs considerable economic costs, both immediately after the first occurrence and over the subsequent 5 years. Work absence and inpatient admissions are major drivers of costs, with drug and outpatient visits only contributing to a small part of the total healthcare costs associated with ASCVD. ASCVD is a common condition, and its effects on individuals, society and healthcare systems should be a key consideration in treatment decision-making and healthcare policy. ASCVD is strongly linked to modifiable risk factors , 4, 8, aAdditional file 1: Supplementary Methods. Supplementary Table S1. Data sources and variables included. Supplementary Table S2. ICD-10 codes for the broad definition of ASCVD. Supplementary Table S3. ICD-10 codes and hospital procedure codes (KVÅ) selected from the National Patient Register. Supplementary Table S4. ATC codes selected from the National Prescribed Drug Register. Supplementary Table S5. Numbers of people in the control group who met the criteria for ASCVD in each year of the study. Supplementary Table S6. Cumulative total, direct and indirect cost data for individuals with ASCVD and control individuals. Supplementary Table S7. Mean annual direct costs (€) before and after the first observed occurrence of ASCVD in 2011.


LPL. By combining data from African Americans, European Americans, and West Africans, we identify three conditionally independent variants with varying amounts of ancestrally differentiated allele frequencies. Using out-of-sample data, we demonstrate improved prediction achievable by accounting for multiple causal variants and locus-specific ancestry effects at a single locus.The vast majority of human populations and individuals have mixed ancestry. Consequently, adjustment for locus-specific ancestry is essential for genetic association studies. To empower association studies for all populations, it is necessary to integrate effects of locus-specific ancestry and genotype. We developed a joint test of ancestry and association that can be performed with summary statistics, is independent of study design, can take advantage of locus-specific ancestry effects to boost power in association testing, and can utilize association effects to fine map admixture peaks. We illustrate the test using the association between serum triglycerides and  LPL from African Americans, European Americans, and West Africans, we identify three conditionally independent variants with varying amounts of ancestrally differentiated allele frequencies.We describe a joint test of ancestry and association calculated from summary statistics from admixture mapping and association studies. Using data for serum triglycerides and  The idea of admixture mapping as a gene mapping tool was described in 1954,,,,A union-intersection test (UIT) involves the composite null hypothesis that all null hypotheses are true against the composite alternative hypothesis that at least one alternative hypothesis is true.,,,An intersection-union test (IUT) involves a composite null hypothesis that at least one null hypothesis is true against the composite alternative hypothesis that all alternative hypotheses are true.LPL (MIM: 609708) for serum triglycerides.,Here, we derive an IUT that is a joint ancestry and association test. This test is applicable for any study design and phenotype that can be analyzed using a generalized linear mixed model. Furthermore, this test is based on signed Z score for a locus-specific ancestry effect and let Z score for an association effect of genotype conditional on locus-specific ancestry. The product of these two Z scores is the test statistic for the joint test of ancestry and association. The product can only be nonzero if both Z scores are nonzero. The product follows the product normal distribution. We used the R package RMediationAt a marker, let Z scores convey useful information that would be lost in a test based on The signs of the The definition of admixture as the process of intermating between previously isolated populations involves, at a minimum, three populations  and two timescales. In our study, the descriptor African American refers to descendants of the Middle Passage. Admixed African Americans resulted from intermating over the last few hundred years. The descriptors African and European refer to the two primary source populations. These two descriptors are intentionally broad and inclusive in order to reflect the genetic diversity in both sources. The timescale of isolation encompasses the tens to hundreds of thousands of years since the split of anatomically modern humans into lineages that predated human dispersal out of Africa.We integrated individual-level data from eight genetic epidemiology studies comprising 23,643 African Americans : the Ath,Serum triglyceride (mg/dL) measurements for HUFS and AADM were made on fasting samples and determined enzymatically, as previously reported.For each study, we followed the same genotype cleaning and imputation steps. First, markers that were strand ambiguous or monomorphic were removed. Using PLINK, we then filtered for individual-level missingness of 0.1, per-marker missingness of 0.05, and a Hardy-Weinberg equilibrium p value of For each of the eight studies of African Americans, we inferred locus-specific ancestry using RFMix.Principal-component analysis of the African data revealed that the top principal component separated Ghanaians from Nigerians.th and 5th degree relatives) in GCTA.To test the effect of locus-specific ancestry on phenotype, we regressed the phenotype on locus-specific ancestry using a linear mixed model, adjusted for age, sex, and study as fixed effects and the relatedness matrix estimated from locus-specific ancestry as a random effect . To reduce confounding by shared environmental effects among close relatives, unrelated individuals were extracted using --grm-cutoff 0.046875 , an approach robust to the unknown genetic mode of inheritance.We generated a genetic score accounting for the effects of age, sex, and genetic variants. For each genetic variant, we weighted the allelic dosage by the estimated allelic effect size. We used the nonoverlapping set of individuals excluded upon filtering for relatedness.Z score for a locus-specific ancestry effect and let Z score for an association effect of genotype conditional on locus-specific ancestry. The product of these two Z scores is a test statistic for the joint test of ancestry and association. Under the null hypothesis, the test statistic is distributed as the product of two standard normal random variables. Conditioning association testing on locus-specific ancestry leads to the property that the two random variables are conditionally independent. By incorporating ancestral allele frequencies into the test, we constrain the test to be one tailed. To establish the validity of this test, we generated 1,000,000 statistical replicates of three independent random variates from the standard normal distribution and calculated the test statistic by multiplying the sign of the first random variate by the product of the other two random variates. For a significance level To integrate locus-specific ancestry and association effects, we derived an IUT to specifically test the composite alternative hypothesis of both locus-specific ancestry and association effects. Let as valid A. On an as valid B. Also aas valid B.Figure We next benchmarked the joint ancestry and association test relative to the standard association test C. We perLPL for serum triglycerides,LPL is the only gene in this interval.Given prior evidence for a locus-specific ancestry effect at We next tested the association of genotype on serum triglycerides at all 1,679 variants in the 184.6 kb interval in 14,895 unrelated African Americans . A totalWe further tested the association of genotype on serum triglycerides at 1,623 variants in 2,083 West Africans and 916 variants in 13,416 unrelated European Americans and combined all results in meta-analysis. Of 1,878 total variants, associations at 302 were genome-wide significant . The mosA) associated with higher serum triglycerides was higher in the European background (68.9%) than in the African background (42.9%) among African Americans  a locus-specific ancestry effect but not an association effect, (2) an association effect but not a locus-specific ancestry effect, or (3) a locus-specific ancestry effect and an association effect. Consequently, these tests are joint tests of ancestry or association, not joint tests of ancestry and association. In contrast, we describe a joint test of ancestry and association with a composite alternative hypothesis of a locus-specific ancestry effect and an association effect. Our test is constructed as a product of locus-specific ancestry and association effects rather than as a sum. The use of a structured association test ensures that the association effect is conditionally independent of the locus-specific ancestry effect. Our test uses a third piece of information, the allele frequencies in the ancestries, to determine whether the sign of the association effect is consistent with the locus-specific ancestry effect. Consequently, our test can leverage the presence of a locus-specific ancestry effect to boost power to detect association while fine mapping the locus-specific ancestry effect by ruling out variants with combinations of allele frequencies and association effects inconsistent with the locus-specific ancestry effect. Joint tests based on sums of LPL such that an increasing amount of European ancestry correlated with higher serum triglycerides . For rs3208305, Africans have more of the allele associated with lower triglycerides, a pattern consistent with the locus-specific ancestry effect. For rs3289 and rs117199990, Africans have more of the allele associated with higher serum triglycerides. Thus, these two associations are inconsistent with the locus-specific ancestry effect. Consequently, the joint test of ancestry and association detected rs3208305 but not rs3289 or rs117199990. Although the associations at rs3289 and rs1177199990 cannot be detected by the joint test, they can be detected using a standard test of association. All three variants have been reported previously to be associated with relevant phenotypes: rs3208305 with postprandial triglyceride levels,,We observed a locus-specific ancestry effect at LPL. We detected no associations at coding variants. The top index variant, rs3208305, tags rs13702 (NC_000008.11:g.19966981T>C), with strong linkage disequilibrium (LD) in both Africans , phs000285.v3.p2 (CARDIA), phs000284.v1.p1 (CFS), phs000007.v32.p13 (FHS), phs001238.v2.p1 (GENOA), phs000286.v4.p1 (JHS), phs000209.v13.p3 (MESA), phs000433.v1.p1 (SIGNET), and phs000200.v12.p3 (WHI). The HUFS and AADM datasets are available from C.N.R. upon reasonable request.


Herpes B virus is a biosafety level 4 pathogen and widespread in its natural host species, macaques. Although most infected monkeys show asymptomatic or mild symptoms, human infections with this virus can cause serious neurological symptoms or fatal encephalomyelitis with a high mortality rate. Herpes B virus can be latent in the sensory ganglia of monkeys and humans, often leading to missed diagnoses. Furthermore, the herpes B virus has extensive antigen crossover with HSV, SA8, and HVP-2, causing false-positive results frequently. Timely diagnosis, along with methods with sensitivity and specificity, are urgent for research on the herpes B virus. The lack of a clear understanding of the host invasion and life cycle of the herpes B virus has led to slow progress in the development of effective vaccines and drugs. This review discusses the research progress and problems of the epidemiology of herpes B virus, detection methods and therapy, hoping to inspire further investigation into important factors associated with transmission of herpes B virus in macaques and humans, and arouse the development of effective vaccines or drugs, to promote the establishment of specific pathogen-free (SPF) monkeys and protect humans to effectively avoid herpes B virus infection. The SARularis) . The Natularis) . Due to Experimental monkeys that satisfy market demand, also known as specific pathogen-free (SPF) monkeys, must be tested negative for mycobacterium tuberculosis (TB), simian retrovirus D (SRV), cercopithecine herpesvirus type 1 (B virus), simian T lymphotropic virus type 1 (STLV-1) and simian immunodeficiency virus (SIV) . AccordiHerpes B virus was first identified in 1932, The International Committee on Taxonomy of Viruses (ICTV) termed it Cercopithecine herpesvirus 1 (CHV-1) in 1999 , which wThis review focuses on summarizing the recent progress and characteristics of viral biology and clinical epidemiology of herpes B virus, and discussing the current and future of the diagnosis, prevention and treatment of herpes B virus, hoping to inspire researchers to solve the existing hazards of herpes B virus, more effectively eliminate B virus from infected groups, and help to prevent its re-infection into established SPF NHP colonies.22.1L) and the short unique region (US). UL is bounded by a terminal repeat region (TRL) and an inverted internal repeat region (IRL), and US is similarly bounded by a terminal repeat region (TRS) and an inverted internal repeat region (IRS)  , 15. Bason (IRS) , while 6on (IRS) , and 70.on (IRS) .134.5 (RL1), which encodes a neurovirulence factor ICP34.5  showed that the genome of BVcy is different from the rhesus macaque B virus isolate (BVrh) E2490 . Ohsawa 2.2The B virus was found to be an enveloped particle with a diameter of about 200 nm, comprised of an electron dense nucleus containing viral DNA within an icosapentahedral capsid encased in an amorphous tegument protein layer, and a lipid envelope coated with viral glycoproteins , 15 (FiThe degree of amino acid homology between B virus and HSV varies from 26.6% (US5) to 87.7% (UL15), and the three least-conserved proteins in B virus are US4, US5 and US12. Twelve glycoproteins are known to be present in the B virus, namely gB, gC, gD, gE, gG, gH, gI, gJ, gK, gL, gM and gN . Homolog2.3Macaca fuscata), (II) cynomolgus macaques isolates, and (III) isolates from pig-tailed macaques (Macaca nemestrina).There is no consensus on the genotyping of the B virus. Smith et al.  suggesteMacaca silenus) does not belong to the three reported genotypes and may represent the fourth genotype of the B virus. Ohsawa et al.  (Cebus apella) , which are usually latent and asymptomatic or cause only mild disease. Significant lesions of the tongue and lips were found only in 332 (2.3%) of 14,400 rhesus monkeys examined by Keeble et al. . Howeverglectus) , capuchi apella) , patas m apella) , which mInfection with the B virus is usually mild and self-limited in Asian macaques. Nevertheless, in some cases, diseases similar to human HSV infection may occur . Although the presence of herpetic skin lesions means the active transmission, B virus-infected macaques without visible mucosal lesions can still transmit the virus. Some slight symptoms may subside within 10-14 days without scarring, occasionally conjunctivitis of varying severity, and no genital tract damage symptoms. But B virus infection in monkeys is lifelong, because the virus can be in the trigeminal ganglia and lumbosacral nerve for a long time –60. DuriB virus infection is common in macaques and the incidence rises with age. It is mainly transmitted through mating, scratching or biting. In sexually mature non-SPF rhesus macaques, the incidence of B virus infection is low, but it rapidly increased after sexual maturity at 3-4 years old, approaching 80-90% B virus-positivity in some groups –67. Duri3.2Human infections are typically caused by monkey bites or scratches, contamination injuries caused by monkey experiment supplies , or exposure to mucosal infectious substances  –71. ExpoThe Centers for Disease Control and Prevention (CDC) claimed that about 50 cases of the B virus were recorded globally from 1932 to 2019, mostly caused by exposure to laboratory or captive macaques and their tissues . In NoveMoreover, the threat to humans from the B virus in global non-laboratory settings has not been studied. A perplexing feature of this zoonotic pathogen infection is that despite B virus-positive macaques having substantial-close interaction and direct exposure with humans, there were few fatal or clinically obvious B virus infections. Before the human infection with B virus in China, fatal cases of B virus have only occurred in the US and Canada following contact with captive macaques. Researchers are similarly baffled by this regional constraint of zoonotic B virus infection.44.1in vitro include Vero cells, Hela cells and other well-established epithelial cell lines . During active infection, B virus DNA can be detected in samples such as saliva or genitourinary samples by PCR tests Figure 4To improve laboratory diagnosis of B virus infections, Scinicariello et al.  developePerelygina et al.  designedPCR tests can be used for quality testing in monkey colonies. Nevertheless, a significant limitation of PCR or virus isolation is that, because of the incubation period of the B virus infection, the diagnostic test would be false-negative, meaning that the presence of viral DNA could not be detected. A positive result occurs only when a latent viral infection is reactivated and the virus sheds in oral and genital secretions. Therefore, serological testing has a higher sensitivity and utility than PCR or virus isolation, which is why it is better suited for screening purposes rather than routine B virus detection. Apart from conventional PCR, real-time PCR and LAMP, digital PCR (the third-generation PCR technology) , PCR amp4.3Typically, serological diagnosis aims to detect anti-B virus antibodies, usually using enzyme-linked immunosorbent assay (ELISA), dot-immunobinding assay (DIA), and enzyme immunoassay (EIA), etc. ELISA is the standard first-line diagnostic test for B virus infection Figure 4In 1986, Katz et al.  establisIn addition to the use of envelope glycoprotein as a detection antigen, synthetic polypeptides have also been employed as an alternative antigen. Perelygina et al.  found thMoreover, monoclonal antibodies recognize the viral antigen to detect B virus. Cropper et al.  preparedHowever, the serological diagnosis could not specifically identify the virus that was actually infected in humans or macaques. According to the positive results of B virus antibody assays, it can only be determined that the macaques were infected with B virus, but not the current status. Lees et al.  measuredPrevious studies have revealed that viral DNA is not necessarily detectable in B virus-positive macaques by PCR, and the virus is latent , 118, ma55.1The relative rarity of zoonotic B virus infections does not provide a financial incentive for the development of drugs that specifically target the B virus. Therefore, the drugs used to treat HSV infection are also referred to treat patients with suspected B virus infection Table 1,If any signs or symptoms of B virus infection or a positive B virus test result exist, intravenous anti-viral therapy is required instead of oral drugs for post-exposure prophylaxis. If CNS symptoms are present, high-dose intravenous ganciclovir is the recommended first choice (5 mg/kg q12h) . If CNS In some cases, the effectiveness of human infection intervention was monitored by suppressing peripheral viral shedding and in others by reducing cerebrospinal fluid antibodies or viral DNA load , 82, 1265.2in vitro supported the idea that high-dose human γ-globulin might be useful in postexposure immunoprophylaxis against B virus infection ] of the National Institutes of Health initiated experimental research contracts to establish and maintain SPF colonies, so far, many countries have built monkey colonies free of the B virus, but non-SPF monkeys are still widely used for research. Latent infection and intermittent reactivation of B virus make it quite difficult to detect infected monkeys accurately, and the antigen crossover of B virus and other pathogens also increases the difficulty of confirming B virus infection. These suggest that it is impossible to completely eliminate the possibility of B virus infection in humans.In addition, there is no vaccine or specific neutralizing antibody drug available to prevent B virus infections. Although acyclovir or ganciclovir can be effective against the B virus, they cannot eradicate it entirely. In accordance with the standardized and secure operating procedures for experiments, therefore, prevention and fortification of personal protection are of paramount importance. When bitten or scratched by a monkey, the wound should be handled immediately, detected and treated in time, even if it is an SPF monkey. It is not possible to take chances, even if the number of individuals infected with the B virus is far lower than the number of persons scratched and bitten by monkeys. Since the B virus may remain dormant in the human body, false-negative tests may conceal serious health risks.The biological characteristics of the B virus have been relatively well studied, but the pathogenesis and immune response to the B virus in macaques and humans need to be further studied. The specific diagnostic methods, medications and vaccines also require deeper investigation and development. Only by preventing well, establishing rapid and accurate diagnostic methods, and developing efficient and feasible treatment methods can it be possible to eliminate the risk of human mortality caused by herpes B virus infection.JL: Writing – original draft, Writing – review & editing. YL: Writing – review & editing. JS: Funding acquisition, Writing – review & editing. LG: Writing – review & editing.


Patients with diabetes mellitus (DM) are always required to be able to control a healthy lifestyle throughout their life to avoid various diseases that can accompany the previous illness. However, psychological factors in the form of despair due to lack of hope make people with diabetes more depressed and less able to control behavior and maintain blood sugar stability, so an excellent internal locus of control is needed to be stronger. This study aimed to determine the effect of hope therapy in reducing hopelessness and increasing internal locus of control in people with DM. The research design used a experimental study with ten randomly selected respondents divided into two groups, namely the control group and the experimental group. Data retrieval using the locus of control scale and the beck hopelessness scale. Data analysis used non-parametric analysis, namely the Mann Whitney test, Wilcoxon test, and Spearman’s Rank Correlation test. The results of the Mann-Whitney U test on the internal locus of control variable show a value of 0.000 and a p score of 0.008 (p <0.05), it can be concluded that there are differences in the internal locus of control in the experimental group compared to the internal locus of control in the control group. The hopelessness variable shows a value of 0.000 and a p score of 0.008 (p <0.05), this indicates that there is a difference in hopelessness in the experimental group and the control group. There is a decrease in hopelessness and an increase in internal locus of control in people with DM given hope therapy. Health is everyone’s need, both physical and mental health; with a person’s mental health, other aspects of life will work optimally , 2. One The high number of people with DM is mostly caused by lifestyle changes, such as lack of physical activity, incorrect eating patterns, poor sleep habits, high levels of stress, negative habits, and difficulty adapting to the environment , 6.People with DM must constantly control their lifestyle for life to avoid various risks of psychological. Some research results show that people with DM experience psychological problems –8. PatieOne of the important factors for controlling lifestyle is locus of control which is an individual’s belief in finding the power that controls life, especially internal locus of control where the power that controls his life comes from the individual himself .Previous research has shown that several things can improve the internal health locus of control on compliance behavior and self-care behavior of patients with DM, as well as on dietary compliance and adherence to the rules of life for people with DM , 11. EffIndividuals with an external locus of control that relate their health progress and decline due to fate or luck will tend to feel that their efforts are not producing results. In contrast, individuals with an internal locus of control are more willing to follow advice and input to treat their disease because they believe that their disease progresses depends on the management of oneself , 16. TheInternal locus of control is very important and needed for dm sufferers because it can control behavior to obey the rules so as to improve the quality of life and health , 20, unfOne individual factor that does not have an internal locus of control is a sense of hopelessness; if hopelessness in people with DM can be reduced, it can increase the internal locus of control. The relationship between the internal locus of control and hopelessness is significant. However, the association between the internal locus of control and hopelessness and the discussion of the relationship between the two is still minimal. , 29, so Individuals who have low expectations tend to benefit the most from an expectation intervention –32. HopeHope is the most important factor for people with DM, because with the hope that people with DM can balance the psychological or physical pressures of the disease they suffer . ConcernThis study aimed to determine the effect of hope therapy on reducing hopelessness and increasing internal locus of control in people with DM. This research is expected to provide a theoretical contribution to psychology, especially regarding the internal locus of control and hopelessness in people with DM. The benefits of research can help people with DM to reduce feelings of hopelessness and increase internal locus of control to keep patients away from the prolonged negative impact of their disease. Health workers and related people with DM can be used as helpful information during the treatment process.Locus of control was invented by Rotter in the 1960s , Locus oThe cause of the internal or external locus of control is due to the experience related to the failure he experienced, and this factor can sometimes be confused regarding the behavior given to the experience. Some people can have different interpretations of reinforcement and rewards offered by others –43.The concept of locus of control is defined as an individual’s belief in locating the forces that control life. Internal locus of control is defined as the individual views the power that controls his life that comes from the individual himself in being responsible for what happens during his life . IndividNegative and repetitive life events can contribute to the formation of hopelessness because it will lead gradually to the belief that there is powerlessness in the face of stressors. Individuals who respond to negative events form three dimensions of causal attribution, namely internal to external, stable to unstable, and global to specific . The devHopelessness theory describes that the three negative inferential forces function as a cognitive diathesis that cannot be associated with the possibility of increasing hopelessness or depression. On the other hand, when faced with negative events, only individuals with cognitive diathesis are at greater risk of hopelessness and depression because this hopelessness theory is a stress diathesis depression model. In summary, individuals with cognitive diathesis should not be at risk for hopelessness or depression in the presence of positive events or the absence of negative events –50.Hopelessness is understood as a core element of negative expectations about an unappreciated outcome or feeling powerless to change the likelihood of that outcome . HopelesHope therapy states that a person’s emotions follow the desired goal; when thoughts are directed to a positive realm, emotions will reflect as they think. People with low expectations tend to think negatively about their desires, impacting their confidence and ability to achieve their dreams , 52.Hope is a force that can indeed function as an essential therapy for change by taking advantage of current expectations. Some of the principles that support hope therapy, including the fact that hope therapy is designed to be a short and semi-structured form of treatment in Indonesia whose primary focus is the current target .In addition, to increase hope, the therapist helps clients to focus on goals, possibilities, and past successes rather than on problems or failures. There are four main components in hope therapy, namely, Finding Hope, Bonding Hope, Raising Hope, and Reminding Hope .The components in hope therapy are divided into four; namely, Hope Finding is a search for hope and is owned by every client to be built and developed as a therapeutic process. Every individual has a different way of wishing, and various expectations can be in the form or purpose . The secPeople with DM tend to experience symptoms of depression due to their illness; one of the symptoms of depression that is the main trigger is hopelessness. Despair makes it difficult for people with DM to internalize their disease to help their recovery. People with DM who experience hopelessness tend to have an external locus of control. Hence, people with DM feel that their efforts to recover are futile because they put their hopes and actions on the outside –59. In dThe general expectation that the outcome of an event depends on the individual’s abilities and efforts is called the internal locus of control  and idenThus it can be concluded that when hopelessness decreases, the internal locus of control will increase, to reduce despair, hope therapy is given as a form of positive psychological intervention that focuses on improving hope.Hopelessness decreases through hope therapy and internal locus of control increases in people with diabetes mellitus.Research proposals are presented before the ethics committee to ensure the impact of the research to be carried out, while ensuring that it does not violate the code of ethics. The results of the study by the Experimental Research Ethics Commission of the Faculty of Psychology, University of Muhammadiyah Malang, approved the research . In addition, each participant is also required to provide willingness and commitment (informed consent) during the intervention process. The authors assured the participants that participant data would be kept confidential.This study uses a experimental type of research with a control group experimental design pre-test-posttest design; in this design, there are two groups selected randomly, then given a pretest to determine the initial state of the experimental group and the control group. After being manipulated, the dependent variable was re-measured with the same measuring instrument in two groups with two different situations.The participants in this study were people with DM in the Bareng Village area of Malang City Indonesia as many as 57 people, but after screening and asking for willingness, only 10 people were willing to be participants so that the ten people with DM were divided into two groups, five people as an experimental group and five people as a randomly selected control group. Participants between the ages of 30–50.Hopelessness was measured using the Beck Hopelessness scale (BHS) which was adapted from research by Sarfika  as many The research procedure begins with a literature review. It determines research methods that are relevant to the research topic, followed by searching for respondents who are following the characteristics of research respondents, one of the characteristics is that respondents are willing and committed to participating in the program as well as internal locus of control scores and the Beck hopelessness scale in the low category. (pre-test data). Ten respondents matched the characteristics of the research subjects; then, a random assignment process was carried out to determine the experimental and control groups. The experimental group was given intervention or treatment in the form of hope therapy.Hope therapy is carried out in 12 intervention sessions for 30–45 minutes, which consists of; 1) Hope finding (1 session), that is, providing knowledge about expectations and finding out and exploring the desires and expectations of the subject based on aspects, 2) Hope bonding (2 sessions), that is, fostering a strong and hopeful alliance or cooperation with the subject, 3) Hope enchancing (4 sessions), that is, increasing hopeful thinking on the subject and providing feedback related to the subject’s thoughts regarding expectations, 4) Hope reminding (3 sessions), that is, teaching the subject how to monitor himself by using their thoughts about the expectations that have been made before and making plans after the therapy has been completed, 5) Termination (2 sessions), i.e. closing the intervention by drawing conclusions from the intervention sessions that have been carried out, looking at the difference between the initial condition and the last condition after the intervention is carried out and the administration of posttests using the internal locus of control and beck hopelessness scale .The implementation of the intervention, in the form of hope therapy, was carried out in the experimental group. No treatment was carried out in the control group by the intervention module that had been prepared and tested. After the intervention process was carried out, measurements were taken (post-test data) to see the effect of hope therapy on hopelessness and internal locus of control.After the intervention process is complete and the data is collected, data analysis is carried out using non-parametric data from the Mann-Whitney and Wilcoxon Test. Mann Whitney test to test the hypothesis; if the significance value > 0.05, then H0 is accepted, and Ha is rejected. If the significance value is <0.05, then H0 is rejected, and Ha is accepted. Meanwhile, Wilcoxon to determine the difference between the results of the pretest and post-test in both control and experimental groups or to test the effectiveness of the therapy with the help of the SPSS version 23 for windows program. In addition to quantitative data, interviews were conducted before and after treatment to determine differences in psychological conditions that could not be obtained through a scale as additional data that was considered essential for the psychological well-being of respondents, as well as an evaluation of the intervention process.Based on the results of the study, a description of the study subjects consisting of 10 participants . As for more details as in The results showed that the comparison of pretest and posttest on internal locus of control variables in each group to answer the research hypothesis using the Wilcoxon test because there were too few study subjects and abnormal data, as in In The Mann-Whitney test was performed on both variables to see the differences in internal locus of control and hopelessness in the experimental and control groups. The results of the Mann-Whitney U test on the internal locus of control variable show a value of 0.000 and a p score of 0.008 (p <0.05), it can be concluded that there are differences in the internal locus of control in the experimental group compared to the internal locus of control in the control group. This shows that Ha is proven, namely that there is a difference in the internal locus of control between the control and experimental groups, where the internal locus of control in the experimental group has changed scores compared to the control group. The hopelessness variable shows a value of 0.000 and a p score of 0.008 (p <0.05), this indicates that there is a difference in hopelessness in the experimental group and the control group, which supports the proof of Ha, namely the difference in despair between the two groups, namely the experimental and control groups.Spearman Rank correlation analysis was used to determine the relationship between the two variables, namely despair and internal locus of control. It is known that the correlation coefficient of the hopelessness variable on the internal locus of control is -0.937 with a p-value of 0.000 (p<0.05), and the direction of the relationship is negative. Thus, it can be concluded that hopelessness negatively correlates with an internal locus of control. That is, the lower one’s despair, the higher the internal locus of control one has.In addition to the results of the quantitative analysis, guided interviews were also conducted through items on the internal locus of control scale and the hopelessness scale. The following is additional data obtained through the results of interviews between before and after the treatment was given, it can be seen in Tables The results showed a significant increase in the internal locus of control by reducing hopelessness using hope therapy in people with DM. The experimental group that was given this intervention had lower hopelessness than the control group, so an increase in the internal locus of control occurred significantly in the experimental group. The results showed an increase in hopelessness in the control group, although the increase was not significant even though no treatment was given. This proves that the control group that is not given any treatment can increase despair if it is not handled correctly, which means it is essential to provide treatment to hopeless people.The existence of the role of hopelessness in influencing a person’s locus of control is explained through previous research that a person’s sense of hopelessness does occur due to the influence of last negative events. Still, when the feeling of hopelessness can be controlled, it will change the control center in internalizing an event –69.Despair is very vulnerable to occur in people who suffer from DM because DM is a chronic disease and lasts a lifetime, so it can affect how the sufferer views his or future .Hopelessness is negative energy that a person has, especially for people with unpleasant events such as suffering from DM. When one is hopeless and has no hope, it will be difficult for a person to have a locus of control. Binzer explained that negative energy, such as loss of motivation, can contribute to a low internal locus of control and vice versa to provide control over himself and tend to depend on the circumstances around him .Based on research, Hope therapy can effectively reduce hopelessness in people with DM. Changes occurred in several sessions conducted with aspects of hopelessness and the presence of other factors, namely support from family, which decreased despair in each session. In the therapeutic process, the experimental group has a higher level of hopelessness than the control group. Therefore it is essential to pay more attention to the barriers and supports for people with DM in the experimental group.This research still has limitations in terms of time for therapy because it was carried out during the Covid-19 pandemic, resulting in researchers having limited time and space due to health protocol regulations and considering the safety of research subjects.Based on the research that has been done, it can be concluded that there is a decrease in hopelessness and an increase in internal locus of control in people with DM given hope therapy. In addition, research subjects can also understand that the illness they suffer can be controlled and is a personal responsibility that can be handled alone. The implications of this research are expected to bring awareness to people with DM to keep hope, not give up on their illness, and be able to internalize negative events in life and not surrender or surrender to fate.


Malassezia is the most abundant fungus on human skin and is associated with a variety of conditions, including atopic eczema, atopic dermatitis, dandruff, folliculitis, pityriasis versicolor, and seborrheic dermatitis. Here, we conducted mycovirome studies on 194 public transcriptomes of Malassezia  screened against all available viral proteins. Transcriptomic data were assembled de novo resulting in 1,170,715 contigs and 2,995,306 open reading frames (ORFs) that were subsequently tracked for potential viral sequences. Eighty-eight virus-associated ORFs were detected in 68 contigs from 28 Sequence Read Archive (SRA) samples. Seventy-five and thirteen ORFs were retrieved from transcriptomes of Malassezia globosa and Malassezia restricta, respectively. Phylogenetic reconstructions revealed three new mycoviruses belonging to the Totivirus genus and named Malassezia globosa-associated-totivirus 1 (MgaTV1); Malassezia restricta-associated-totivirus 1 (MraTV1) and Malassezia restricta-associated-totivirus 2 (MraTV2). These viral candidates extend our understanding of the diversity and taxonomy of mycoviruses as well as their co-evolution with their fungal hosts. These results reflected the unexpected diversity of mycoviruses hidden in public databases. In conclusion, this study sheds light on the discovery of novel mycoviruses and opens the door to study their impact on disease caused by the host fungus Malassezia and globally, their implication in clinical skin disorders. Mycoviruses are viruses that selectively infect and multiply in fungal cells.  Sclerotinia sclerotiorum  CP aa identity finding in the tick metagenome. Despite the higher identity percentage, we suggest the genome be given the provisional name Malassezia restricta-associated-Totivirus 2 (MraTV2). We hypothesised that the occurrence of this virus in the tick metagenome could be associated with Malassezia colonization/infection of the ticks. Notably, this is the first identification of the viruses described above in Malassezia. The third virus found in Malassezia restricta belongs to a species previously described as MrV40L [In the family us genus . Based ot 83.69–8.62% CP aTotivirus, Giardiavirus, Victorivirus and unclassified Totivirus genera, we used maximum-likelihood phylogenetic analyses of the RdRP and CP amino acid sequences (Totivirus), we assumed that these candidate viruses belong to the Totiviridae family. Phylogenetic analysis performed using RdRps sequences showed a clustering of the newly found MgaTV1 and MraTV1 with mycoviruses of unclassified Totivirus and Totivirus genus. The MrV40L identified in this study was clustered with the previously described MrV40L by Park et al. as expected  sequences exhibited more than 50% identity with the closest known Totivirus. Nonetheless, the criteria for defining a new virus species were met, because both the CP sequences and the other two polyproteins (RT-like superfamily) shared less than 50% identity. Therefore, we considered this virus as a new species. The MraTV2 sequences shared more than 80% identity with DTtV1 (UYL95681.1), which was identified in a tick metagenome. We have therefore hypothesised that the tick could harbor Malassezia on its surface or in its digestive system, and that this virus comes from Malassezia rather than from the tick itself. However, this hypothesis cannot be tested because the metagenomic data are not publicly available. Our RNA-Seq approach also successfully identified both RdRp and CP sequences of MrV40L in the M. restricta meta transcriptomic data of the strain from which this virus had been characterised [Malassezia genome. On the other hand, the 86% Malassezia transcriptomes in which we did not detect any virus-associated contigs might be infected by unknown mycoviruses. Indeed, omics approaches cannot detect unknown viruses that are too dissimilar from currently known viral sequences.Although the presence of dsRNA and virus-like particles has been reported in some  species ,17, dataeviously , we condiviruses , thus methe ICTV . The CP Totivirus-like contigs clustered with unclassified Totiviridae, unclassified Totivirus, and the genus Totivirus. Regarding the RdRP amino acid sequences of the two predicted Totivirus-like contigs, MgaTV1 and MraTV1 clustered with an unclassified Totivirus that is known to infect sea cucumbers (Holothuroidea and Echinodermata) [Erysiphe necator, the agent of powdery mildew in grapes.The phylogenetic analysis of the detected RdRP and CP protein sequences showed that the amino acid sequences of the three newly identified dermata)  and consTotivirus, totivirus sp., and those of MraTV1 clustered with an unclassified Totiviridae that has been detected in a tick metagenome. CP sequences of MraTV2 clustered with Dali Totiv tick virus 1 which was also found in a tick metagenome. Both RdRP and CP amino acid sequences of MrV40L from M. restricta were closely related and shared significant similarity with MrV40L  identified by Park et al. [Regarding ORFs encoding the CP, those of MgaTV1 clustered with the unclassified k et al.  (Figure Malassezia associated-mycoviruses identified in this study raises questions about their potential influence on Malassezia virulence. Viruses from the genus Totivirus are currently known to infect a variety of fungi, in some of which a decrease in virulence has been observed [Totiviridae mycoviruses on Malassezia yeast. They showed that MsMV1 induces the overexpression of transcription factors and ribosomal genes and a higher level of IFN-β expression in cultured macrophages. Meanwhile, MrV40L has also been shown to upregulate TLRs and cytokines and to have an impact on several physiological processes in M. restricta [Malassezia species. The identification of a virus by only an in silico approach does not demonstrate that the virus infects a living organism. This must be confirmed by other methods.The presence of observed . In addiestricta . It is pM. globosa and M. restricta, which are among of the most abundant yeast of the human skin mycobiome. Additionally, we report for the first time the detection of three novel Totiviridae signatures in Malassezia transcriptomes. These findings expand our knowledge about the diversity of mycoviruses in this yeast of clinical importance.In conclusion, this study confirms the presence of mycoviruses in


Enamovirus-like virus were discovered. This is the first report of GAMaV, GCSV, GLRaV4, GPGV, GRGV, GRVFV, and GSyV-1 infecting grapevines in Mexico; the impact of these pathogens on production is unknown.This is the first viral metagenomic analysis of grapevine conducted in Mexico. During the summer of 2021, 48 plants displaying virus-like symptoms were sampled in Queretaro, an important grapevine-producing area of Mexico, and analyzed for the presence of viruses via high-throughput sequencing (HTS). The results of HTS were verified by real-time RT-PCR following a standardized testing scheme (Protocol 2010). Fourteen different viruses were identified, including grapevine asteroid mosaic-associated virus (GAMaV), grapevine Cabernet Sauvignon reovirus (GCSV), grapevine fanleaf virus (GFLV), grapevine fleck virus (GFkV), grapevine Pinot gris virus (GPGV), grapevine red globe virus (GRGV), grapevine rupestris stem pitting-associated virus (GRSPaV), grapevine rupestris vein feathering virus (GRVFV), grapevine Syrah virus 1 (GSyV-1), grapevine virus B (GVB), and grapevine leafroll-associated viruses 1, 2, 3, 4 . Additionally, divergent variants of GLRaV4 and GFkV, and a novel  Vitis spp.) are one of the world’s most economically important horticultural crops, used for both food and wine production [Grapevines  is a revolutionary molecular technique for plant virus diagnostics . The techttps://www.avq.com.mx/; accessed on 1 June 2023). The quality of this wine has been recognized internationally, as evidenced by multiple awards  in international competitions .In Mexico, there are limited studies on plant viruses using HTS. These studies include the detection of viruses in citrus trees, berries, beans, and non-cultivated plants ,14,15,16Enamovirus-like virus.Grapevines displaying virus-like symptoms were collected from commercial vineyards and analyzed via HTS. A confirmatory test of putative viral sequences was provided by real-time RT-PCR following a standardized testing scheme. Findings include the first report of grapevine asteroid mosaic-associated virus (GAMaV), grapevine Cabernet Sauvignon reovirus (GCSV), grapevine leafroll-associated virus 4 (GLRaV4), grapevine Pinot gris virus (GPGV), grapevine red globe virus (GRGV), grapevine rupestris vein feathering virus (GRVFV), and grapevine Syrah virus 1 (GSyV-1) infecting grapevines in Mexico, and the discovery of divergent variants of GLRaV4 and GFkV, and a new In the summer of 2021, 48 grapevine leaf samples were collected from 13 commercial vineyards located across the State of Queretaro, Mexico. The climate in Queretaro is semi-arid at an elevation of 1859 m above sea level and an average annual precipitation of 570 mm. At the time, although some plants were asymptomatic, others presented foliar and fruit necrosis, dwarfism, leaf spotting, curling, reddening, yellowing and chlorosis . DiverseFor sample preparation, 0.5 g of leaf petioles were cut from the grapevine samples and flash frozen in liquid nitrogen before grinding. Quick-RNA Plant Miniprep Kit  spin columns were used according to the manufacturer’s protocol. RNA concentration and quality was determined using Qubit  and Qsep100 , for which the required parameters were as follows: RQN > 8; minimum RNA amount per sample = 500 ng.Library construction of nucleic acids was carried out with the TruSeq Stranded Total RNA with Ribo-Zero Plant Kit , followed by the assembly of the obtained cDNA libraries after repairing their ends or converting them for ligation with the adapters by unique dual-indexes. Finally, sequencing was performed in an Illumina NovaSeq 6000 , with a flow cell SP Reagent Kit (2 × 100 cycles), located at the National Genomic Sequencing Laboratory Tec-BASE .E-value < 1 × 10−5) were further inspected to confirm their likely viral homology and host range based on their closest hits in GenBank.Illumina bcl2fastq2 software was used to perform demultiplexing and adapter trimming. Later, de novo assembly was performed using SPAdes software . To annoThe putative proteins and potential open reading frames (ORFs) encoded by virus like contigs determined by HTS were identified using ORFfinder and subsequent BLASTp annotation. To identify conserved domains within these proteins, the Pfam database  was searAlignments of the sequences of viruses here identified and sequences from GenBank with the highest identity were carried out in MEGAX . Brieflyhttps://fps.ucdavis.edu/fgr2010.cfm; accessed on 1 June 2023). These molecular tests have been described previously [Following the viral metagenomic analysis, real-time RT-PCR assays were employed to confirm the presence of known viruses. Such assays are part of Protocol 2010, and they are used routinely at Foundation Plant Services  as part of the grapevine certification program  as per the manufacturer’s specifications. Each reaction  included 2 µL of RNA and final primer and probe concentrations of 900 and 250 nM, respectively. In addition, virus-specific assays were multiplexed with an 18S rRNA assay to confirm presence of RNA .Forty-eight grapevine plants originating from commercial vineyards located in Queretaro, Mexico were screened for viruses via HTS . The paiMetagenomic analysis identified sequences of several diverse viruses and viroids in the transcriptome of both symptomatic and asymptomatic sampled grapevines. Not a single plant was free of viral agents, though the composition of the infecting viruses was diverse. Of the total read counts, the majority of sequence belongs to the host plant; only 0.02–2% reads mapped to viruses or viroids.Known viruses and viroids were identified in the analyzed plants as single or mixed infections . VirusesThe BLASTn annotation of the assembled contigs revealed a genetically diverse set of sequences from viruses known to infect grapevine. We identified the genomes of 14 viruses, represented by 116 distinct isolates  among the 48 samples we analyzed. The nucleotide identity of our sequence to the closest virus homolog in GenBank is provided in As indicated in Enamovirus, family Solemoviridae). The new viral sequence displayed an average depth of 3411 reads . ORF 2-encoded protein  has the highest similarity to the GEV1 RdRp , and it is produced by ribosomal slippage. A CP with a molecular weight of 21.86 kDa is encoded by ORF 3 (197 aa), based on 85% aa identity (100% query coverage) with the corresponding product of GEV1; additionally, a luteo coat super family (F62-N195) motif was also identified. Lastly, the ORF 5 545 aa-long protein (readthrough protein) is related to an aphid transmission protein of GEV1 (60.24 kDa) and shares 90% aa identity (100% query coverage) with the ortholog protein expressed by the GEV2 genome. Finally, phylogenetic analysis confirmed the relation of GEV2 with other enamoviruses, including GEV1 , was determined to be 6288 nucleotide (nt) long (GenBank: OR066156). BLASTn comparisons revealed the closest homologous sequence as GEV1 isolate CS-BR with 87.91% nt identity (98% query coverage). The GEV2 genome  containsing GEV1 .Real-time RT-PCR was used to confirm the presence of grapevine viruses identified by HTS. Virus detection was then validated by comparing HTS and real-time RT-PCR results for each sample. The real-time RT-PCR assays produced Ct values that ranged from 20 to 39 .Enamovirus was discovered. The high incidence of viral agents calls for an improvement in the sanitation protocols and management of grapevine viruses in Mexico. Previously, we discussed the risk posed by viral pathogens for grapevine production in this country [In this study, we completed the first viral metagenomic analysis of grapevine in Mexico. HTS results were validated using a standardized virus detection protocol, which involves real-time RT-PCR assays. A set of 48 grapevine plants, the majority displaying virus-like symptoms, was found to be infected by 14 viruses and 5 viroids; additionally, a new potential member of the genus  country . Some ofGrapevine plants analyzed during this study were collected from different commercial vineyards along the State of Queretaro. Overall, the average age of these vineyards was 8 years old. Mexico lacks nurseries and propagation programs with large-scale operations; thus, most of the stock planting material originated from abroad. The European origin of some of these plants may explain the presence of viruses linked to serious economic losses in the old world, like GFLV and GPGV ,30,31. OWe compared our virus sequences to reference genomes available in public databases to determine genetic diversity. The most divergent viruses were GFkV, GLRaV4, GRGV, and GRVFV. In the case of GFkV and GLRaV4, phylogenetic analysis suggests that these novel variants do not belong to any previously known type , and instead represent potentially new subclades. Thus, the near-complete genome sequences for these viruses create the first base-line framework for major grapevine viruses and their genetic variants identified in Mexico.Enamovirus infecting this host [https://ictv.global/report/chapter/solemoviridae/solemoviridae/enamovirus; accessed on 1 May 2023). Consequently, based on sequence identity, the putative novel virus found in sample “Vitis-TrPADL13” represents a new species belonging to the genus Enamovirus, which we provisionally name GEV2. Given the presence of additional viral agents in this sample, it was not possible to ascertain whether GEV2 is associated with symptoms. Complementary experiments on its pathogenicity are needed, likewise, studies investigating the distribution of this new virus.In 2017, GEV1 was described infecting grapevine in Brazil, being the first report of a virus in the genus his host . CriteriThis work establishes the first report of virus and viroid populations infecting grapevine in Queretaro. Similar field surveys should be performed in other important grapevine growing regions in Mexico, such as Sonora, Zacatecas and Coahuila. By contrast, initial efforts have already been done in Aguascalientes and Baja California through small-scale surveys ,35. EpidAccording to our results, mixed infections were common among the analyzed samples, being GRSPaV frequently detected. Initially, GRSPaV was considered the causal agent of Rupestris stem pitting, a common graft-transmitted disease. However, subsequent studies have shown that GRSPaV does not affect growth or develop symptoms in cultivars such as Albano, Madeleine Sylvaner, Ortega, and Savagnin rose . In addiAlthough the grapevine industry is relatively small in Mexico, some of the oldest vineyards in the American continent are situated in the country, being important sources of genetic diversity that must be preserved. As a result, the first search for grapevine viruses via HTS was conducted in Mexico. To our knowledge, this is the initial report of GAMaV, GCSV, GLRaV4, GPGV, GRGV, GRVFV, and GSyV-1 infecting grapevines in Mexico; in all cases, detection was confirmed by two independent analyses . To prevent future negative impacts on the local grapevine industry, new plantations should prioritize the use of certified clean stock, with virus diagnosis and tissue culture as requirements, to provide the means for long-term elimination of viral diseases from vineyards.


Early treatment of RA improves clinical outcomes; however, the impact on health economic outcomes is unclear. This review sought to investigate the relationship between symptom/disease duration and resource utilization/costs and the responsiveness of costs following RA diagnosis.A systematic search was performed on Pubmed, EMBASE, CINAHL and Medline. Studies were eligible if patients were DMARD-naïve and fulfilled 1987 ACR or 2010 ACR/EULAR RA classification criteria. Studies had to report symptom/disease duration and resource utilization or direct/indirect costs as health economic outcomes. The relationships between symptom/disease duration and costs were explored.Three hundred and fifty-seven records were identified in a systematic search; nine were eligible for analysis. The mean/median of symptom/disease duration in studies ranged between 25 days and 6 years. Annual direct costs of RA following diagnosis showed a U-shaped distribution in two studies. Longer symptom duration before starting a DMARD (>180 days) was associated with lower health-care utilization in the first year of RA diagnosis in one study. Annual direct and indirect costs 6 months before RA diagnosis were higher in patients with shorter symptom duration (<6 months) in one study. Given the clinical and methodological heterogeneities, the association between symptom/disease duration and costs after diagnosis was not computed.The association between symptom/disease duration at the time of DMARD initiation and resource utilization/cost in patients with RA remains unclear. Health economic modelling with clearly defined symptom duration, resource utilization and long-term productivity is vital to address this evidence gap. Key messagesThe association between symptom/disease duration before DMARD initiation and health economic outcomes in RA is unclear.Clinical and methodological heterogeneities impede direct comparison of health economic outcomes across RA studies.Longitudinal studies with defined symptom duration and long-term RA-associated costs will address this research question.The impact of early treatment on clinical outcomes in RA is well reported . HoweverTherefore, as a first step, the relationship between symptom/diagnosis duration at the time of DMARD initiation and subsequent resource utilization/costs needs to be identified. We sought to investigate this through a systematic review of cost-of-illness and cost-effectiveness studies of DMARD-naïve RA patients.Rheumatology Advances in Practice online.The full Methods section is detailed in https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42017077593.The protocol was registered on the International Prospective Register of Systematic Reviews (PROSPERO 2017 CRD42017077593); Rheumatology Advances in Practice online). Additional records were identified through independent manual database searching, external sources and reference scanning of relevant retrieved full-text articles. Study selection, data extraction and quality assessment were done independently by two authors (I.S. and R.S.); discrepancies were resolved by consensus or through a third reviewer (A.Bo.). PubMed, EMBASE, CINAHL and Medline electronic databases were searched up to 25 January 2023. All systematic searches were conducted using the same search terms and strategy  checklist  and a moA meta-analysis/regression on the association between disease/symptom duration and costs could not be performed owing to the number of studies and methodological heterogeneity, especially in reporting of health economic outcomes. Cost data per patient per year for the reported duration in studies were recorded and summarized in a unifying currency of US Dollars 2021 after adjusting for the Purchasing Power Parity (PPP) and Consumer Price Index (CPI) 2021 , 7.Nine articles were included in this systematic review. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) flow chart shows the literature search results .Rheumatology Advances in Practice online. Cost categories, source of cost reference and results in local currency are summarized in Rheumatology Advances in Practice online.Sociodemographic and clinical characteristics of patients are summarized in The symptom, disease or diagnosis duration variables reported at baseline varied. Two studies reported symptom duration , 14, sixResource utilization and cost data across studies were heterogeneous . Three sDirect medical costs were reported in six studies  was recorded in four studies , 14, 15.Study perspective refers to the point of view adopted in the economic evaluations , i.e. whRheumatology Advances in Practice online.Quality assessment has been included in et al. . Cumulative 5-year productivity cost loss (HCA) was highest in the TCZ plus MTX group (€51 700) compared with the TCZ monotherapy and MTX monotherapy groups . After adjusting for PPP and CPI 2021, total direct health-care-related costs (mean) in USD 2021 at the end of year 1 were $15 546, $8350 and $17 840 per patient for the TCZ plus MTX, TCZ and MTX groups, respectively. The association between symptom duration and health-care or productivity costs in the overall cohort or by treatment groups was not assessed.Verhoeven et al.  reportedet al. . The mean number of visits to medical specialists peaked at 6 months in the RA group . In addition, RA patients with disease duration ≥10 years from the Utrecht RA Cohort study group were included to capture costs data for patients with longstanding RA. There was a significant difference in annual direct costs between the four groups. The median annual direct costs per patient showed a U-shaped distribution, i.e. costs were high for patients with follow-up duration 0 to ≤2 years (€2923) and reduced after 2–6 years (€1967), but increased again for ≥10 years follow-up duration (€3778). Data from the group with the shortest follow-up duration were extracted for Verstappen et al.  assessedet al.  6 months before diagnosis in patients with disease duration <6 months compared with ≥6 months were $240/month ± $285 and $144/month ± $149, P < 0.001, respectively. Likewise, indirect costs were higher in patients with a disease duration <6 months as opposed to ≥6 months  at baseline. After adjusting for PPP and CPI 2021, the annual mean total direct and indirect costs 6 months before diagnosis per person in USD 2021 were $12 663 for <6 months and $7174 for ≥6 months groups. Overall, annual direct and indirect costs 6 months before RA diagnosis were higher in patients with shorter symptom duration (<6 months).Newhall-Perry et al.  assessedet al. [van Jaarsveld et al.  assessedA number of studies were excluded because study participants could receive at least one DMARD before study enrolment . Tables This study highlighted several interesting findings. Firstly, two studies reported a U-shaped distribution of costs over disease duration following an RA diagnosis. Total costs were high during the initial years, slightly lower thereafter, then high again for a disease duration of ≥5 years  and >10 Secondly, functional disability was a predictor of productivity costs in three studies , 10, 12.One study reported that the annual direct and indirect costs 6 months before diagnosis were higher in those with a symptom duration of <6 months before the start of DMARD therapy compared with those with a symptom duration ≥6 months . In contvs 6 years) also differed across studies (In this review, we could not delineate the aggregated-level data related to the relationship between symptom/disease/diagnosis duration and cost categories owing to the heterogeneity of the following factors: timing and duration of data collection regarding resources and costs; type of resources/cost-categories reported; and inconsistency in reported disease, symptom or diagnosis duration . Moreove studies .Before the era of early treatment, RA costs were related to established disease. Patients had more frequent hospitalization  and moreClear definitions of RA onset and duration have been proposed , becauseA strength of this review is the broad range of health economic outcomes and types of health economic studies that were included. Both direct and indirect costs, and cost-of-illness and cost-utility studies were within the scope of this review. Observational and clinical trials were also included.However, only a small number of studies fulfilled our strict inclusion criteria. In addition, studies that enrolled patients who had recently been treated with DMARDs before study recruitment were not included in this review. Furthermore, meta-analyses/regression were not possible owing to the different types of health economic outcomes reported.This review is the first to highlight a vital evidence gap in early arthritis: what is the financial consequence of diagnosing and treating patients with RA during the early disease phase? Health economic modelling with carefully defined symptom duration, resource utilization, treatment and long-term productivity costs is vital to address this important question.rkad040_Supplementary_DataClick here for additional data file.


We describe the detection of epizootic hemorrhagic disease virus (EHDV) serotype 8 in cattle farms in Sardinia and Sicily in October–November 2022. The virus has a direct origin in North Africa; its genome is identical (>99.9% nucleotide sequence identity) to EHDV serotype 8 strains detected in Tunisia in 2021. Orbivirus and circulate in multiple serotypes (Culicoides spp. biting midges. Bluetongue primarily affects sheep and in recent decades has been described multiple times in the European Union (EU), causing devastating repercussions on animal trade  lists epizootic hemorrhagic disease (EHD) as a disease of wild and domestic ruminants caused by EHD virus (EHDV). EHDV is related to bluetongue virus (BTV), the etiologic agent of bluetongue, a disease of ruminants. Both viruses belong to the genus On October 25, 2022, respiratory distress, erosions of the muzzle and oral mucosa, and drooling were reported in 3 cattle at farm 1, located near the city of Trapani, Sicily ; serum ahttps://www.thermofisher.com). We developed a real-time reverse transcription PCR (rRT-PCR) specific for the S2 of the EHDV-8 strain detected in Tunisia in 2021 (EHDV-8 TUN 2021) because the available test designed for the S2 segment of the EHDV-8 reference serotype  did not detect EHDV-8 TUN 2021 (https://www.innovative-diagnostics.com), and by virus neutralization . Genotyping confirmed the presence of EHDV-8 TUN 2021–like strains. We reported the outbreak to Italy’s Ministry of Health, which notified WOAH and the European Commission, which imposed animal movement restrictions within a 150-km radius of the outbreak sites. We selected 1 EHDV-8–positive blood sample from Sardinia for WGS; results confirmed that the Sardinia EHDV-8 strain (GenBank accession nos. OP897265–74) shares high nucleotide sequence identity (>99.9%) with multiple EHDV-8 TUN 2021–like strains. WGS of the Sicily strains is ongoing. We isolated the virus from all rRT-PCR–positive blood samples; all serum samples tested positive by ELISA and by virus neutralization (antibody titer 10–20).Orbivirus incursion into the EU sustained by EHDV-8 was predictable, considering the distribution of this virus in Tunisia and likely in neighboring countries (Confirmation of novel