Recommendations of the DNA Commission of the International Society for Forensic Genetics (ISFG) on short tandem repeat sequence nomenclature.

The DNA Commission of the International Society for Forensic Genetics (ISFG) has developed a set of nomenclature recommendations for short tandem repeat (STR) sequences. These recommendations follow the 2016 considerations of the DNA Commission of the ISFG, incorporating the knowledge gained through research and population studies in the intervening years. While maintaining a focus on backward compatibility with the CE data that currently populate national DNA databases, this report also looks to the future with the establishment of recommended minimum sequence reporting ranges to facilitate interlaboratory comparisons, automated solutions for sequence-based allele designations, a suite of resources to support bioinformatic development, guidance for characterizing new STR loci, and considerations for incorporating STR sequences and other new markers into investigative databases.

Neuron-specific enolase as a prognostic biomarker in acute ischemic stroke patients treated with reperfusion therapies.

Introduction: Ischemic stroke is a significant global health concern, with reperfusion therapies playing a vital role in patient management. Neuron-specific enolase (NSE) has been suggested as a potential biomarker for assessing stroke severity and prognosis, however, the role of NSE in predicting long-term outcomes in patients undergoing reperfusion therapies is still scarce. Aim: To investigate the association between serum NSE levels at admission and 48 h after reperfusion therapies, and functional outcomes at 90 days in ischemic stroke patients. Methods: This study conducted a prospective cross-sectional analysis on consecutive acute ischemic stroke patients undergoing intravenous fibrinolysis and/or endovascular thrombectomy. Functional outcomes were assessed using the modified Rankin Scale (mRS) at 90 days post-stroke and two groups were defined according to having unfavorable (mRS3-6) or favorable (mRS0-2) outcome. Demographic, clinical, radiological, and laboratory data were collected, including NSE levels at admission and 48 h. Spearman's coefficient evaluated the correlation between analyzed variables. Logistic regression analysis was performed to verify which variables were independently associated with unfavorable outcome. Two ROC curves determined the cut-off points for NSE at admission and 48 h, being compared by Delong test. Results: Analysis of 79 patients undergoing reperfusion treatment following acute stroke revealed that patients with mRS 3-6 had higher NIHSS at admission (p < 0.0001), higher NIHSS at 24 h (p < 0.0001), and higher NSE levels at 48 h (p = 0.008) when compared to those with mRS 0-2. Optimal cut-off values for NSE0 (>14.2 ng/mL) and NSE48h (>26.3 ng/mL) were identified, showing associations with worse clinical outcomes. Adjusted analyses demonstrated that patients with NSE48h > 26.3 ng/mL had a 13.5 times higher risk of unfavorable outcome, while each unit increase in NIHSS24h score was associated with a 22% increase in unfavorable outcome. Receiver operating characteristic analysis indicated similar predictive abilities of NSE levels at admission and 48 h (p = 0.298). Additionally, a strong positive correlation was observed between NSE48h levels and mRS at 90 days (r = 0.400 and p < 0.0001), suggesting that higher NSE levels indicate worse neurological disability post-stroke. Conclusion: Serum NSE levels at 48 h post-reperfusion therapies are associated with functional outcomes in ischemic stroke patients, serving as potential tool for patient long-term prognosis.

Patients' and Providers' Perspectives of the Transition of Care from Hospital to Community for Hispanic/Latino Adults with Diabetes.

INTRODUCTION: Transition of care is a critical point of care for Hispanic/Latino patients with diabetes, who face higher rates of diabetes-related complications and hospital readmission and challenges during the transition of care from hospital to the community. METHODS: Using semi-structured interviews, with questions tailored to each group, we explored patients' and providers' perspectives of their experiences and challenges during the transition of care of Hispanic/Latino adult patients with diabetes from the hospital to the community. RESULTS: Overlapping themes emerged from both patient and provider interviews identifying common barriers regarding the transition of care, including discordance due to language/communication barriers, perceived burden/nuisance, lack/unawareness of available resources, and lack of patient education. CONCLUSIONS: Findings in this study provide a frame of reference that can be used to address challenges facing Hispanic/Latino patients with diabetes. The uniqueness of this study is its exploration of the perspectives of patients and healthcare providers and the overlapping themes that emerged. Additionally, timely outpatient follow-up care, education, and reducing communication discordance can help with lowering readmission rates for patients with diabetes, especially those who are medically disadvantaged and have language barriers.

A systems thinking framework for understanding rising childhood obesity in the Caribbean.

OBJECTIVE: The aim of this study is to develop a systems thinking framework to describe the common complexities of childhood obesity in the Caribbean region and identify potential areas of intervention. METHODS: Group model building (GMB) is a form of systems science. Trained GMB facilitators in Puerto Rico, the US Virgin Islands, Barbados, and Trinidad and Tobago convened a group of multi-disciplinary stakeholders in a series of virtual meetings in 2021 to elaborate a hypothesis of the system driving childhood obesity represented by causal loop diagrams (CLD). Commonalities and differences between the CLDs from each island were identified and reconciled to create a synthesized CLD. RESULTS: A single explanatory CLD across the islands was developed and includes nine reinforcing loops. These loops addressed the interconnected role of schools, policy, commercial determinants, community and the personal experience of the child in rising childhood obesity rates. CONCLUSIONS: Despite differences across settings, there is a core system driving childhood obesity in the Caribbean, as described by stakeholders in GMB workshops. Policy solutions to the problem must be multi-faceted and multi-level to address the interlinked reinforcing loops of the complex system and reduce rates of childhood obesity.

Discrepancies in the results reported for multiple sclerosis clinical trials: A comparison between ClinicalTrials.gov and peer-reviewed journals.

OBJECTIVE: We aimed to compare the results of phase III and IV clinical trials examining drugs to treat multiple sclerosis (MS) registered at ClinicalTrials.gov to those published in peer-reviewed journals. METHODS: After identifying trials registered at ClinicalTrials.gov, consecutive searches were conducted in PubMed, EMBASE and Google Scholar for matching publications. Information regarding participants and efficacy and safety results was extracted and compared. The degree of consistency was classified as 'concordant', 'discrepant' or 'not comparable'. The Kaplan-Meier method was used to model time to reporting. RESULTS: In total, 65 trials were appraised. The median time from completion to reporting was shorter for ClinicalTrials.gov (16.4 vs 27.3 months; p = 0.010). Information availability was generally higher in journals except for serious adverse events (SAEs) (86.2% vs 100.0%, p = 0.029) and their description (78.2% vs 100.0%, p < 0.001). However, 45 trials had at least one reporting discrepancy (69.2%). Three studies omitted one or more primary outcomes in the matching journal publication. Regarding safety results, the lowest consistencies were found for causes of death (60.0%) and description of SAEs (27.9%). CONCLUSION: Consulting both ClinicalTrials.gov and journals increases the accessibility to MS clinical trial results. Some data were frequently missing or disagreed between sources, raising concerns about transparency and generalizability of results.

Facilitated Peer Group Mentoring for Underrepresented Biomedical Researchers: Facilitators' Experiences and Implications for Dissemination of a Curriculum.

Peer group mentoring facilitated by senior faculty represents an effective approach. However, for underrepresented biomedical researchers, access to senior faculty from underrepresented racial/ethnic groups is limited. We explored motivations, benefits, and challenges for facilitators enrolled to deploy an intervention in the context of a randomized controlled trial that tested two peer group mentoring strategies for underrepresented early career researchers. Peer group sessions were co-facilitated by two senior underrepresented faculty. Thirty-six faculty were recruited as facilitators over four years. The facilitators' primary motivation was advancing the diversity of the workforce, the primary benefit was satisfaction from helping underrepresented researchers, and the primary challenge was time. Understanding motivations, benefits, and challenges of facilitators informs efforts in recruiting and retaining facilitators and disseminating this curriculum and others like it, to the broader community.

Discovery of Novel, Selective, and Nonbasic Agonists for the Kappa-Opioid Receptor Determined by Salvinorin A-Based Virtual Screening.

Modulating the kappa-opioid receptor (KOR) is a promising strategy for treating various human diseases. KOR agonists show potential for treating pain, pruritus, and epilepsy, while KOR antagonists show potential for treating depression, anxiety, and addiction. The diterpenoid Salvinorin A (SalA), a secondary metabolite of Salvia divinorum, is a potent and selective KOR agonist. Unlike typical opioids, SalA lacks a basic nitrogen, which encouraged us to search for nonbasic KOR ligands. Through structure-based virtual screening using 3D pharmacophore models based on the binding mode of SalA, we identified novel, nonbasic, potent, and selective KOR agonists. In vitro studies confirmed two virtual hits, SalA-VS-07 and SalA-VS-08, as highly selective for the KOR and showing G protein-biased KOR agonist activity. Both KOR ligands share a novel spiro-moiety and a nonbasic scaffold. Our findings provide novel starting points for developing therapeutics aimed at treating pain and other conditions in which KOR is a central player.

ß-D-Glucose-Reduced Silver Nanoparticles Remodel the Tumor Microenvironment in a Murine Model of Triple-Negative Breast Cancer.

Breast cancer is the most diagnosed type of cancer worldwide and the second cause of death in women. Triple-negative breast cancer (TNBC) is the most aggressive, and due to the lack of specific targets, it is considered the most challenging subtype to treat and the subtype with the worst prognosis. The present study aims to determine the antitumor effect of beta-D-glucose-reduced silver nanoparticles (AgNPs-G) in a murine model of TNBC, as well as to study its effect on the tumor microenvironment. In an airbag model with 4T1 tumor cell implantation, the administration of AgNPs-G or doxorubicin showed antitumoral activity. Using immunohistochemistry it was demonstrated that treatment with AgNPs-G decreased the expression of PCNA, IDO, and GAL-3 and increased the expression of Caspase-3. In the tumor microenvironment, the treatment increased the percentage of memory T cells and innate effector cells and decreased CD4+ cells and regulatory T cells. There was also an increase in the levels of TNF-α, IFN-γ, and IL-6, while TNF-α was increased in serum. In conclusion, we suggest that AgNPs-G treatment has an antitumor effect that is demonstrated by its ability to remodel the tumor microenvironment in mice with TNBC.

Understanding Health Care Students' Perceptions, Beliefs, and Attitudes Toward AI-Powered Language Models: Cross-Sectional Study.

BACKGROUND: ChatGPT was not intended for use in health care, but it has potential benefits that depend on end-user understanding and acceptability, which is where health care students become crucial. There is still a limited amount of research in this area. OBJECTIVE: The primary aim of our study was to assess the frequency of ChatGPT use, the perceived level of knowledge, the perceived risks associated with its use, and the ethical issues, as well as attitudes toward the use of ChatGPT in the context of education in the field of health. In addition, we aimed to examine whether there were differences across groups based on demographic variables. The second part of the study aimed to assess the association between the frequency of use, the level of perceived knowledge, the level of risk perception, and the level of perception of ethics as predictive factors for participants' attitudes toward the use of ChatGPT. METHODS: A cross-sectional survey was conducted from May to June 2023 encompassing students of medicine, nursing, dentistry, nutrition, and laboratory science across the Americas. The study used descriptive analysis, chi-square tests, and ANOVA to assess statistical significance across different categories. The study used several ordinal logistic regression models to analyze the impact of predictive factors (frequency of use, perception of knowledge, perception of risk, and ethics perception scores) on attitude as the dependent variable. The models were adjusted for gender, institution type, major, and country. Stata was used to conduct all the analyses. RESULTS: Of 2661 health care students, 42.99% (n=1144) were unaware of ChatGPT. The median score of knowledge was "minimal" (median 2.00, IQR 1.00-3.00). Most respondents (median 2.61, IQR 2.11-3.11) regarded ChatGPT as neither ethical nor unethical. Most participants (median 3.89, IQR 3.44-4.34) "somewhat agreed" that ChatGPT (1) benefits health care settings, (2) provides trustworthy data, (3) is a helpful tool for clinical and educational medical information access, and (4) makes the work easier. In total, 70% (7/10) of people used it for homework. As the perceived knowledge of ChatGPT increased, there was a stronger tendency with regard to having a favorable attitude toward ChatGPT. Higher ethical consideration perception ratings increased the likelihood of considering ChatGPT as a source of trustworthy health care information (odds ratio [OR] 1.620, 95% CI 1.498-1.752), beneficial in medical issues (OR 1.495, 95% CI 1.452-1.539), and useful for medical literature (OR 1.494, 95% CI 1.426-1.564; P<.001 for all results). CONCLUSIONS: Over 40% of American health care students (1144/2661, 42.99%) were unaware of ChatGPT despite its extensive use in the health field. Our data revealed the positive attitudes toward ChatGPT and the desire to learn more about it. Medical educators must explore how chatbots may be included in undergraduate health care education programs.

Managing Placenta Accreta and Massive Hemorrhage: A Case Report on Anesthetic and Surgical Interventions.

Obstetric haemorrhage is a leading cause of maternal morbidity and mortality and is a common reason for intensive care unit (ICU) admission in the postpartum. Primary postpartum obstetric haemorrhage is associated with four main causes: tone, thrombin, trauma, and tissue. Regarding the last one, placenta accreta is an abnormal invasion of the placenta into the myometrium. Early diagnosis of placenta accreta allows for better perioperative management; however, it is sometimes only identified during caesarean delivery when the placenta cannot be removed. We report a case of a 37-year-old woman with a history of caesarean section due to placenta previa, who was admitted at 36 weeks and 1 day for an urgent caesarean section (c-section) due to cord presentation. A subarachnoid block (SAB) was used for anaesthesia. It was chosen over general anaesthesia because it allows the patient to experience the birth of her children, enhances pain control, and avoids complications associated with general anaesthesia. Besides our centre has expertise in neuraxial anaesthesia. During the procedure, placental accretism and massive haemorrhage occurred, and a life-saving abdominal hysterectomy was needed. The patient experienced hypotension, partially responsive to volume replacement and vasopressors, leading to norepinephrine infusion and conversion to general anaesthesia. The surgery lasted 2.5 hours with a blood loss of 3500 ml. The patient was extubated without complications and transferred to the post anaesthesia care unit (PACU). Risk factors for placenta accreta spectrum (PAS) include previous surgery and placenta previa with a prior c-section. Antenatal diagnosis is crucial, and women with risk factors should undergo imaging at experienced centres. Delivery centres must have protocols for unexpected PAS and major obstetric haemorrhage. Both general and neuraxial anaesthesia can be suitable for managing PAS, and caesarean hysterectomy is often required to control haemorrhage. Postoperatively, adequate monitoring and care is essential. PAS management should involve excellent communication between a multidisciplinary team in specialised centres.

Left Atrial Septal Pouch (LASP) and Cryptogenic Stroke: A Narrative Review.

Over 20% of ischemic strokes are cardioembolic strokes, necessitating research into thrombus formation locations, particularly the left atrial appendage (LAA). The left atrial septal pouch (LASP), which is linked to thrombus development and stasis, has drawn attention recently as a possible thromboembolic location, especially in atrial fibrillation (AF). The primary aim of this review is to explore LASP's role in cryptogenic strokes and to discuss the methods used to assess LAA anatomy. Imaging modalities such as cardiac computed tomography (CT) and transesophageal echocardiography (TEE) are crucial for diagnosing and characterizing LASP. LASP, found in about one-third of individuals, provides an additional site for thrombus development in the left atrium. The potential clinical implications of LASP-related thromboembolic events include the need for targeted therapeutic strategies, such as anticoagulant medication and, in some cases, consideration of LASP closure to prevent recurrent strokes. Further investigation is required to elucidate LASP's involvement in thromboembolic events and to guide stroke prevention in at-risk patients.

The IPEFA model: An initiative for online training and education as applied by the International Society for Forensic Genetics.

The IPEFA model was developed for organizing online training and education events as applied by the International Society for Forensic Genetics (ISFG). It consists of five phases: 1) Input, 2) Preparation, 3) Execution, 4) Feedback, and 5) Assessment. This document details these phases and shows IPEFA's first practical application to the 2023 edition of the virtual ISFG Summer School. Through sharing the experiences, we aim to provide transparency and engage with potential participants and teachers to (virtual) training and education events as organized by the ISFG. The model may also be useful for others organizing (online) events. We have experienced that evaluation of events with input and feedback from both the (potential) participants and teachers is essential for successful training and education. This takes time which is limited in everyone's busy agenda's and may therefore not always be performed with the care it requires. Since these aspects are crucial, however, we aim to keep following the principles as outlined in the IPEFA model.

Pressing issues for oral care quality improvement: findings from the EU DELIVER project.

BACKGROUND: While oral health often takes a backseat to other health domains, it silently affects nearly half of the Worldwide population. The DELIVER project, funded by the EU's Horizon Europe program, seeks to develop a blueprint model for improving the quality of oral health care for everyone. METHODS: Applying the Nominal Group Technique (NGT), 17 stakeholders from various backgrounds participated in identifying pressing issues for oral care quality improvement across practice, community, and policy levels. RESULTS: The results revealed significant differences at the different levels, with accessibility emerging as a prominent issue, encompassing affordability, availability, and acceptability of oral healthcare services. CONCLUSIONS: These findings emphasizes the need for policy reforms, increased investments, and a shift towards preventive and patient-centered dental care practices. It highlights the importance of collaborative efforts with multi-stakeholders and prioritizing pressing issues on a multi-level to drive positive change in improving oral care quality.

Humans seropositive for Trypanosoma cruzi co-infected with intestinal helminths have higher infectiousness, parasitaemia and Th2-type response in the Argentine Chaco.

BACKGROUND: The Gran Chaco ecoregion is a well-known hotspot of several neglected tropical diseases (NTDs) including Chagas disease, soil-transmitted helminthiasis and multiparasitic infections. Interspecific interactions between parasite species can modify host susceptibility, pathogenesis and transmissibility through immunomodulation. Our objective was to test the association between human co-infection with intestinal parasites and host parasitaemia, infectiousness to the vector and immunological profiles in Trypanosoma cruzi-seropositive individuals residing in an endemic region of the Argentine Chaco. METHODS: We conducted a cross-sectional serological survey for T. cruzi infection along with an intestinal parasite survey in two adjacent rural villages. Each participant was tested for T. cruzi and Strongyloides stercoralis infection by serodiagnosis, and by coprological tests for intestinal parasite detection. Trypanosoma cruzi bloodstream parasite load was determined by quantitative PCR (qPCR), host infectiousness by artificial xenodiagnosis and serum human cytokine levels by flow cytometry. RESULTS: The seroprevalence for T. cruzi was 16.1% and for S. stercoralis 11.5% (n = 87). We found 25.3% of patients with Enterobius vermicularis. The most frequent protozoan parasites were Blastocystis spp. (39.1%), Giardia lamblia (6.9%) and Cryptosporidium spp. (3.4%). Multiparasitism occurred in 36.8% of the examined patients. Co-infection ranged from 6.9% to 8.1% for T. cruzi-seropositive humans simultaneously infected with at least one protozoan or helminth species, respectively. The relative odds of being positive by qPCR or xenodiagnosis (i.e. infectious) of 28 T. cruzi-seropositive patients was eight times higher in people co-infected with at least one helminth species than in patients with no such co-infection. Trypanosoma cruzi parasite load and host infectiousness were positively associated with helminth co-infection in a multiple regression analysis. Interferon-gamma (IFN-γ) response, measured in relation to interleukin (IL)-4 among humans infected with T. cruzi only, was 1.5-fold higher than for T. cruzi-seropositive patients co-infected with helminths. The median concentration of IL-4 was significantly higher in T. cruzi-seropositive patients with a positive qPCR test than in qPCR-negative patients. CONCLUSIONS: Our results show a high level of multiparasitism and suggest that co-infection with intestinal helminths increased T. cruzi parasitaemia and upregulated the Th2-type response in the study patients.

Distinctive evolution of alveolar T cell responses is associated with clinical outcomes in unvaccinated patients with SARS-CoV-2 pneumonia.

The evolution of T cell molecular signatures in the distal lung of patients with severe pneumonia is understudied. Here, we analyzed T cell subsets in longitudinal bronchoalveolar lavage fluid samples from 273 patients with severe pneumonia, including unvaccinated patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or with respiratory failure not linked to pneumonia. In patients with SARS-CoV-2 pneumonia, activation of interferon signaling pathways, low activation of the NF-κB pathway and preferential targeting of spike and nucleocapsid proteins early after intubation were associated with favorable outcomes, whereas loss of interferon signaling, activation of NF-κB-driven programs and specificity for the ORF1ab complex late in disease were associated with mortality. These results suggest that in patients with severe SARS-CoV-2 pneumonia, alveolar T cell interferon responses targeting structural SARS-CoV-2 proteins characterize individuals who recover, whereas responses against nonstructural proteins and activation of NF-κB are associated with poor outcomes.

WHO/ICC Classification for Myelodysplastic Neoplasms/Syndromes Performs Better for Subtype Cytomorphological Diagnosis?

The International Consensus Classification of Myeloid Neoplasms and Acute Leukemias (ICC) and the 5th edition of the WHO classification (WHO 2022) have refined the diagnosis of myelodysplastic syndromes (MDS). Both classifications segregate MDS subtypes based on molecular or cytogenetic findings but rely on the subjective assessment of blast cell percentage and dysplasia in hematopoietic cell lineages. This study aimed to evaluate interobserver concordance among 13 cytomorphologists from eight hospitals in assessing blast percentages and dysplastic features in 44 MDS patients. The study found fair interobserver agreement for the PB blast percentage and moderate agreement for the BM blast percentage, with the best concordance in cases with <5% BM blasts and >10% BM blasts. Monocyte count agreement was fair, and dysplasia assessment showed moderate concordance for megakaryocytic lineage but lower concordance for erythroid and granulocytic lineages. Overall, interobserver concordance for MDS subtypes was moderate across all classifications, with slightly better results for WHO 2022. These findings highlight the ongoing need for morphological evaluation in MDS diagnosis despite advances in genetic and molecular techniques. The study supports the blast percentage ranges established by the ICC but suggests refining BM blast cutoffs. Given the moderate interobserver concordance, a unified classification approach for MDS is recommended.

A novel neuroprotective method against ischemic stroke by accelerating the drainage of brain interstitial fluid.

Ischemic stroke is a leading cause of death and disability worldwide. Inflammatory response after stroke determines the outcome of ischemic injury. A recent study has reported an efficient method, epidural arterial implantation (EAI), for accelerating interstitial fluid (ISF) drainage, which provides a promising strategy to clear pro-inflammatory cytokines in the brain extracellular space (ECS). In this study, the method of EAI was modified (m-EAI) to control its function of accelerating the ISF drainage at different time points following ischemic attack. The neuroprotective effect of m-EAI on ischemic stroke was evaluated with the transient middle cerebral artery occlusion (tMCAO) rat model. The results demonstrated the accumulation of IL-1ß, IL-6, and TNF-α was significantly decreased by activating m-EAI at 7 d before and immediately after ischemic attack in tMCAO rats, accompanied with decreased infarct volume and improved neurological function. This study consolidates the hypothesis of exacerbated ischemic damage by inflammatory response and provides a new perspective to treat encephalopathy via brain ECS. Further research is essential to investigate whether m-EAI combined with neuroprotective drugs could enhance the therapeutic effect on ischemic stroke.

Absence of Crimean-Congo hemorrhagic fever virus in wild lagomorphs and their ticks in Spanish Mediterranean ecosystems.

Crimean-Congo hemorrhagic fever virus (CCHFV) is an emerging tick-borne pathogen in different European countries. Since 2013, clinical and fatality cases associated with CCHFV infection have been reported in humans in Spain. During the last few years, endemic circulation of this virus has been detected in ticks and wild ungulates in this country, but the role of other sympatric wild species in the sylvatic cycle of this multi-host virus remains poorly understood. The aims of the present study were to assess exposure to CCHFV in wild lagomorphs in southern Spain, a CCHFV endemic area, and to determine the presence of the virus in ticks feeding on these species. Serum samples from 473 European wild rabbits (Oryctolagus cuniculus) and 162 Iberian hares (Lepus granatensis), and 120 ticks feeding on 85 of these wild lagomorphs were collected on 133 hunting grounds between 2018/2019 and 2021/2022 hunting seasons. The presence of antibodies against CCHFV was assessed in all serum samples using a commercial ELISA, whereas ticks were tested for the presence of CCHFV-RNA by a multiplex RT-qPCR that detects all known genotypes of this virus. None of the 635 (0.0 %; 95 %CI: 0.0-0.6) lagomorphs tested had anti-CCHFV antibodies and CCHFV-RNA was not found in any of the 120 (0.0 %; 95 %CI: 0.0-3.0) ticks analyzed. To the best of the authors knowledge, this is the first epidemiological study conducted on CCHFV in Iberian hare worldwide. Our findings indicate absence of exposure to CCHFV in European wild rabbit and Iberian hare populations, as well as in their ticks, which suggests that they do not seem to play a relevant role in the epidemiology of CCHFV in Mediterranean ecosystems of southern Spain.