Venous thromboembolism (VTE) developing after ankle sprain. Comparison with VTE after knee arthroplasty.

BACKGROUND: Venous thromboembolism (VTE) is a critical complication after non-major trauma or surgery. While the risk and severity of VTE following major orthopedic surgery is well-documented, there is significant knowledge gap regarding, non-major trauma such as ankle sprains. METHODS: We analyzed data from the RIETE registry to assess the clinical characteristics, VTE prophylaxis usage, and outcomes in patients with VTE following ankle sprain versus those post elective knee arthroplasty. We aimed to assess the risk and severity of VTE in a population traditionally considered at lower risk. Risk stratification was performed using the TRiP(cast) score. RESULTS: Among 1,250 patients with VTE, those with ankle sprain (n = 459) were much younger than those post knee arthroplasty (n = 791), less often female, had fewer comorbidities, and received VTE prophylaxis less often (27% vs. 93 %). During anticoagulation, 26 patients developed recurrent VTE, 31 had major bleeding, and 12 died (fatal PE 3, fatal bleeding 2). There were no differences between the two groups in the rates of VTE recurrences (rate ratio (RR): 1.65; 95%CI: 0.69-3.88) or death (RR: 1.12; 95%CI: 0.33-3.46), but patients with VTE after ankle sprain had a lower rate of major bleeding (RR: 0.39; 95%CI: 0.13-0.99). CONCLUSIONS: Ankle sprain patients are often undertreated for VTE prophylaxis and have similar severity of VTE than those undergoing elective knee surgery, indicating the need for a more customized approach to VTE management.

Molecular phylogeny and divergence time of Harpalyce (Leguminosae, Papilionoideae), a lineage with amphitropical diversification in seasonally dry forests and savannas.

Our knowledge of the systematics of the papilionoid legume tribe Brongniartieae has greatly benefitted from recent advances in molecular phylogenetics. The tribe was initially described to include species marked by a strongly bilabiate calyx and an embryo with a straight radicle, but recent research has placed taxa from the distantly related core Sophoreae and Millettieae within it. Despite these advances, the most species-rich genera within the Brongniartieae are still not well studied, and their morphological and biogeographical evolution remains poorly understood. Comprising 35 species, Harpalyce is one of these poorly studied genera. In this study, we present a comprehensive, multi-locus molecular phylogeny of the Brongniartieae, with an increased sampling of Harpalyce, to investigate morphological and biogeographical evolution within the group. Our results confirm the monophyly of Harpalyce and indicate that peltate glandular trichomes and a strongly bilabiate calyx with a carinal lip and three fused lobes are synapomorphies for the genus, which is internally divided into three distinct ecologically and geographically divergent lineages, corresponding to the previously recognized sections. Our biogeographical reconstructions demonstrate that Brongniartieae originated in South America during the Eocene, with subsequent pulses of diversification in South America, Mesoamerica, and Australia. Harpalyce also originated in South America during the Miocene at around 20 Ma, with almost synchronous later diversification in South America and Mexico/Mesoamerica beginning 10 Ma, but mostly during the Pliocene. Migration of Harpalyce from South to North America was accompanied by a biome and ecological shift from savanna to seasonally dry forest.

Identifying patients with prostate cancer at increased risk for haematuria during anticoagulation for venous thromboembolism.

BACKGROUND: Haematuria is a common complication in prostate cancer patients receiving anticoagulation for venous thromboembolism (VTE). Early identification of at-risk patients might help to reduce its incidence and severity. METHODS: We used data from the RIETE registry to develop a prognostic score for haematuria during the first year of anticoagulation for VTE. The prognostic score was built using regression coefficients. RESULTS: From March 2001 through March 2021, 1934 patients with prostate cancer and acute VTE were enrolled. Of these, 1034 (53 %) initially presented as pulmonary embolism and 900 (47 %) as isolated deep vein thrombosis (DVT). During anticoagulation (median 181 days; inter-quartile range: 97-354), 99 patients (5.1 %) developed haematuria (fatal 1, major 27, non-major 72). The incidence rate was: 8 events per 100 patient-years (95%CI 6.5-9.7). Median time to haematuria was 53 days (IQR 4-134). On multivariable analysis, recent haematuria, initial presentation as DVT, comorbidity, metastases, haemoglobin levels <11 g/dL, creatinine >1.2 mg/dL, and radiotherapy independently predicted the risk for haematuria. C-statistics was 0.71 (95%CI: 0.65-0.77). A cut-off of ≥1.5 points classified 312 patients (20 %) at high-risk and had the highest sensitivity (51 %; 95%CI: 39-62) and specificity (82 %; 95%CI: 79-83). Our score improved the performance and non-event net reclassification index (NRI) of the RIETE score (c-statistics: 0.61; 95%CI: 0.54-0.68; NRI: 0.09) or VTE-BLEED score (c-statistics: 0.64; 95%CI: 0.58-0.71; NRI: 0.76). CONCLUSIONS: A prognostic score for haematuria during anticoagulation for VTE performed well in patients with prostate cancer, and improved identification compared to other validated scores.

IgG4-Related Disease: A Rare Case of Simultaneous Lung and Retroperitoneal Involvement.

Immunoglobin G4-related disease is a progressive immune-mediated fibroinflammatory condition that can affect any organ, causing a tumor-like swelling appearance. We present a case of a 57-year-old male who presented with a one-month history of weight loss, constant abdominal pain with dorsal irradiation, night sweats, and respiratory symptoms. CT scan revealed multiple mediastinal and retroperitoneal adenopathies, right pulmonary consolidation, and retroperitoneal fibrosis. Transthoracic pulmonary biopsy and excisional cervical lymph node biopsy revealed fibroinflammatory disease related to IgG4, with normal serum IgG4. The patient presented a good response to glucocorticoids, a clinical characteristic of this disease. The diagnosis of immunoglobin G4-related disease is challenging, due to the nonspecific clinical manifestations, requiring a high level of suspicion in order to perform the appropriate immunohistochemical examination.

A Novel Route to Optimize Placement Equipment Kinematics by Coupling Capacitive Accelerometers.

Machine end-effector kinematic analysis is critical to optimizing transporting components where inertial forces are the main loads. While displacements may be measured with relatively high accuracy in transportation equipment motors, the inertial forces in the transported components are seldom optimized. This is especially relevant in electronic component placement systems, where the components have a wide range of configurations (i.e., geometry and mass) and the deployment dimensional/geometric tolerances are remarkably good. The optimization of these systems requires the monitoring of the real position of the accelerometers relative to the measurement point of interest with sufficient accuracy that allows the assembly position to be predicted instantaneously. This study shows a novel method to calibrate this equipment using triaxial accelerometers on a surface mount machine to measure the end-effector accelerations and velocities in its planar motion. The dynamic equations of the system and the method for integration are presented to address the uncertainty on the exact position of the accelerometer sensors relative to the measuring point of interest exist and allow the position correction to optimize response and accuracy.

Prognostic Impact of Active Cigarette Smoking on Mortality in Patients with Acute Venous Thromboembolic Events, Findings from Real World Data.

Background and Objectives: The influence of smoking habits on mortality, VTE recurrence, and major bleeding in patients receiving anticoagulant therapy for venous thromboembolism (VTE) has not been consistently evaluated. Materials and Methods: We used data from the RIETE (Registro Enfermedad TromboEmbólica) registry to compare mortality, VTE recurrence, and major bleeding risk in smoking versus non-smoking patients with acute VTE. Results: 50,881 patients (43,426 non-smoking and 7455 smoking patients) were included. After a median follow-up of 8.8 months, 7110 patients died (fatal PE 292 and fatal bleeding 281), 3243 presented VTE recurrence, and 1579 had major bleeding. At multivariate analysis, smoking behavior was associated with a higher hazard of death, (HR: 1.28; 95% CI: 1.19-1.40). The risk of VTE recurrence was marginally increased in smoking patients compared to non-smoking patients (1.14; 95% CI: 1.02-1.27). Major bleeding did not differ in smoking and non-smoking patients (1.15; 95% CI: 0.96-1.38). The presence of cancer did not appear to influence the association between smoking habits and death (HR: 1.34; 95% CI: 1.22-1.47 in cancer patients and HR: 1.23; 95% CI: 1.04, 1.45 in non-cancer patients, respectively) Conclusions: the risk of death after an acute episode of VTE appeared to be higher in smoking than in non-smoking patients and this risk is higher between patients presenting PE at the onset of symptoms.

Reading light: leaf spectra capture fine-scale diversity of closely related, hybridizing arctic shrubs.

Leaf reflectance spectroscopy is emerging as an effective tool for assessing plant diversity and function. However, the ability of leaf spectra to detect fine-scale plant evolutionary diversity in complicated biological scenarios is not well understood. We test if reflectance spectra (400-2400 nm) can distinguish species and detect fine-scale population structure and phylogenetic divergence - estimated from genomic data - in two co-occurring, hybridizing, ecotypically differentiated species of Dryas. We also analyze the correlation among taxonomically diagnostic leaf traits to understand the challenges hybrids pose to classification models based on leaf spectra. Classification models based on leaf spectra identified two species of Dryas with 99.7% overall accuracy and genetic populations with 98.9% overall accuracy. All regions of the spectrum carried significant phylogenetic signal. Hybrids were classified with an average overall accuracy of 80%, and our morphological analysis revealed weak trait correlations within hybrids compared to parent species. Reflectance spectra captured genetic variation and accurately distinguished fine-scale population structure and hybrids of morphologically similar, closely related species growing in their home environment. Our findings suggest that fine-scale evolutionary diversity is captured by reflectance spectra and should be considered as spectrally-based biodiversity assessments become more prevalent.

Giardia duodenalis infection in dogs from the metropolitan area of Lisbon, Portugal: prevalence, genotyping and associated risk factors.

ABSTRACT: Giardia duodenalis is a cosmopolitan enteric protozoan that affects a wide range of vertebrates, including humans and dogs. Genetic characterisation reveals eight different assemblages, with A and B having been found mainly in humans and several other animals, and thus considered potentially zoonotic, while C and D are adapted to infect dogs. This study aimed to determine the prevalence of G. duodenalis, their distribution into assemblages, and risk factors associated with their infection of dogs from the metropolitan area of Lisbon. Giardia duodenalis cysts were microscopically identified in 33.8% (27/80) of the faecal samples analysed. Multivariate logistic regression analysis revealed that dogs under 6 months of age and from both breeders and shelters, had a significantly higher risk of being infected with G. duodenalis. Based on phylogenetic analysis of the partial coding sequences for ß-giardin, glutamate dehydrogenase, and triosephosphate isomerase, the parasites found in three dog isolates were typed as G. duodenalis assemblage C, 11 were typed as D, and four were typed as C or D, depending on the targeted genes. The risk to public health seems to be reduced, as no genotypes with zoonotic potential have been detected. Nevertheless, better health management towards a minimisation of the environmental faecal pollution, as well as an increase in the awareness of health professionals, dog owners, dog breeders and caregivers regarding the risks posed by this protozoan to the health of animals and humans, are recommended.

Immunophenotyping of Peripheral Blood, Lymph Node, and Bone Marrow T Lymphocytes During Canine Leishmaniosis and the Impact of Antileishmanial Chemotherapy.

Dogs are a major reservoir of Leishmania infantum, etiological agent of canine leishmaniosis (CanL) a zoonotic visceral disease of worldwide concern. Therapeutic protocols based on antileishmanial drugs are commonly used to treat sick dogs and improve their clinical condition. To better understand the impact of Leishmania infection and antileishmanial drugs on the dog's immune response, this study investigates the profile of CD4+ and CD8+ T cell subsets in peripheral blood, lymph node, and bone marrow of sick dogs and after two different CanL treatments. Two CanL groups of six dogs each were treated with either miltefosine or meglumine antimoniate combined with allopurinol. Another group of 10 clinically healthy dogs was used as control. Upon diagnosis and during the following 3 months of treatment, peripheral blood, popliteal lymph node, and bone marrow mononuclear cells were collected, labeled for surface markers CD45, CD3, CD4, CD8, CD25, and intracellular nuclear factor FoxP3, and T lymphocyte subpopulations were immunophenotyped by flow cytometry. CanL dogs presented an overall increased frequency of CD8+ and CD4+CD8+ double-positive T cells in all tissues and a decreased frequency of CD4+ T cells in the blood. Furthermore, there was a higher frequency of CD8+ T cells expressing CD25+FoxP3+ in the blood and bone marrow. During treatment, these subsets recovered to levels similar to those of healthy dogs. Nevertheless, antileishmanial therapy caused an increase of CD4+CD25+FoxP3+ T cells in all tissues, associated with the decrease of CD8+CD25-FoxP3- T cell percentages. These findings may support previous studies that indicate that L. infantum manipulates the dog's immune system to avoid the development of a protective response, ensuring the parasite's survival and the conditions that allow the completion of Leishmania life cycle. Both treatments used appear to have an effect on the dog's immune response, proving to be effective in promoting the normalization of T cell subsets.

Leaf reflectance spectra capture the evolutionary history of seed plants.

Leaf reflectance spectra have been increasingly used to assess plant diversity. However, we do not yet understand how spectra vary across the tree of life or how the evolution of leaf traits affects the differentiation of spectra among species and lineages. Here we describe a framework that integrates spectra with phylogenies and apply it to a global dataset of over 16 000 leaf-level spectra (400-2400 nm) for 544 seed plant species. We test for phylogenetic signal in spectra, evaluate their ability to classify lineages, and characterize their evolutionary dynamics. We show that phylogenetic signal is present in leaf spectra but that the spectral regions most strongly associated with the phylogeny vary among lineages. Despite among-lineage heterogeneity, broad plant groups, orders, and families can be identified from reflectance spectra. Evolutionary models also reveal that different spectral regions evolve at different rates and under different constraint levels, mirroring the evolution of their underlying traits. Leaf spectra capture the phylogenetic history of seed plants and the evolutionary dynamics of leaf chemistry and structure. Consequently, spectra have the potential to provide breakthrough assessments of leaf evolution and plant phylogenetic diversity at global scales.

Microbiological findings of the maternal periodontitis associated to low birthweight.

OBJECTIVE: To determine the association between the presence of periodontal pathogens and low birthweight. METHODS: This observational and case-control study consisted of mothers of infants weighing <2,500g (Group A), and mothers of newborns weighing ≥2,500g (Group B), born at Hospital da Mulher in Feira de Santana (BA), Brazil. A semi-structured questionnaire covering demographic data, gestational history and aspects related to general and oral health was employed postpartum. Following a complete periodontal examination, biofilm samples were collected at six sites in the mouth. The participants were further categorized in terms of presence or absence of periodontitis. Differences between the groups were determined using Pearson's χ 2 test, odds ratio, and confidence intervals were obtained using the Mantel-Haenszel test. RESULTS: Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Prevotella intermedia were detected by polymerase chain reaction. A total of 303 postpartum women were evaluated, 224 (73.9%) in Group B. Statistically significant differences between the groups were found for age, body mass index and history of previous low birthweight babies. Group A had a higher frequency of periodontitis (33.34%) than Group B (16.22%). P. gingivalis and P. intermedia were detected more frequently among women with periodontitis (74.19% and 88.70%, respectively). CONCLUSION: In this population, there was no association between the presence of maternal periodontal pathogens and the occurrence of low birthweight infants.

Microbiological findings of the maternal periodontitis associated to low birthweight / Achados microbiológicos da periodontite materna associados ao baixo peso ao nascer

ABSTRACT Objective To determine the association between the presence of periodontal pathogens and low birthweight. Methods This observational and case-control study consisted of mothers of infants weighing <2,500g (Group A), and mothers of newborns weighing ≥2,500g (Group B), born at Hospital da Mulher in Feira de Santana (BA), Brazil. A semi-structured questionnaire covering demographic data, gestational history and aspects related to general and oral health was employed postpartum. Following a complete periodontal examination, biofilm samples were collected at six sites in the mouth. The participants were further categorized in terms of presence or absence of periodontitis. Differences between the groups were determined using Pearson's χ 2 test, odds ratio, and confidence intervals were obtained using the Mantel-Haenszel test. Results Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Prevotella intermedia were detected by polymerase chain reaction. A total of 303 postpartum women were evaluated, 224 (73.9%) in Group B. Statistically significant differences between the groups were found for age, body mass index and history of previous low birthweight babies. Group A had a higher frequency of periodontitis (33.34%) than Group B (16.22%). P. gingivalis and P. intermedia were detected more frequently among women with periodontitis (74.19% and 88.70%, respectively). Conclusion In this population, there was no association between the presence of maternal periodontal pathogens and the occurrence of low birthweight infants.

RESUMO Objetivo Determinar a associação entre a presença de patógenos periodontais e o baixo peso ao nascer. Métodos Trata-se de estudo observacional e caso controle. Participaram mães de bebês com peso <2.500g (Grupo A) e de recém-nascidos com peso ≥2.500g (Grupo B) nascidos no Hospital da Mulher, em Feira de Santana, (BA). Dados demográficos, história gestacional e aspectos relacionados à saúde geral e bucal foram obtidos por meio de questionário semiestruturado, aplicado no período pós-parto. Após exame periodontal completo, as amostras de biofilme foram coletadas em seis locais na boca. As participantes foram também categorizadas quanto à presença ou ausência de periodontite. As diferenças entre os grupos foram obtidas pelo teste do χ 2 de Pearson, odds ratio e os intervalos de confiança pelo teste de Mantel-Haenszel. Resultados Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia e Prevotella intermedia foram detectados por reação em cadeia de polimerase. Avaliaram-se 303 mulheres pós-parto, sendo 224 (73,9%) do Grupo B. Diferenças estatisticamente significativas entre os grupos foram observadas quanto à idade, ao índice de massa corporal e a história de partos prévios com bebês de baixo peso ao nascer. O Grupo A apresentou maior frequência de periodontite (33,34%) do que o Grupo B (16,22%). P. gingivalis e P. intermedia foram detectados com maior frequência em mulheres com periodontite (74,19% e 88,70%, respectivamente). Conclusão Nessa população, não houve associação entre a presença de patógenos periodontais maternos e a ocorrência de bebês com baixo peso ao nascer.

Meglumine Antimoniate and Miltefosine Combined With Allopurinol Sustain Pro-inflammatory Immune Environments During Canine Leishmaniosis Treatment.

Canine leishmaniosis (CanL) caused by Leishmania infantum is a zoonotic disease of global concern. Antileishmanial drug therapies commonly used to treat sick dogs improve their clinical condition, although when discontinued relapses can occur. Thus, the current study aims to evaluate the effect of CanL treatments in peripheral blood, lymph node, and bone marrow cytokine profile associated with clinical recovery. Two groups of six dogs diagnosed with CanL were treated with miltefosine combined with allopurinol and meglumine antimoniate combined with allopurinol (MT+A and MG+A), respectively. At diagnosis and after treatment, during a 3-month follow-up, clinical signs, hematological and biochemical parameters, urinalysis results and antileishmanial antibody titers were registered. Furthermore, peripheral blood, popliteal lymph node, and bone marrow samples were collected to assess the gene expression of IL-2, IL-4, IL-5, IL-10, IL-12, TNF-α, TGF-ß, and IFN-γ by qPCR. In parallel, were also evaluated samples obtained from five healthy dogs. Both treatment protocols promoted the remission of clinical signs as well as normalization of hematological and biochemical parameters and urinalysis values. Antileishmanial antibodies returned to non-significant titers in all dogs. Sick dogs showed a generalized upregulation of IFN-γ and downregulation of IL-2, IL-4, and TGF-ß, while gene expression of IL-12, TNF-α, IL-5, and IL-10 varied between groups and according to evaluated tissue. A trend to the normalization of cytokine gene expression was induced by both miltefosine and meglumine antimoniate combined therapies. However, IFN-γ gene expression was still up-regulated in the three evaluated tissues. Furthermore, the effect of treatment in the gene expression of cytokines that were not significantly changed by infection, indicates that miltefosine and meglumine antimoniate combined therapy directly affects cytokine generation. Both combined therapies are effective in CanL treatment, leading to sustained pro-inflammatory immune environments that can compromise parasite survival and favor dogs' clinical cure. In the current study, anti-inflammatory and regulatory cytokines do not seem to play a prominent role in CanL or during clinical recovery.

The role of diversification in community assembly of the oaks (Quercus L.) across the continental U.S.

PREMISE OF THE STUDY: Evolutionary and biogeographic history, including past environmental change and diversification processes, are likely to have influenced the expansion, migration, and extinction of populations, creating evolutionary legacy effects that influence regional species pools and the composition of communities. We consider the consequences of the diversification process in shaping trait evolution and assembly of oak-dominated communities throughout the continental United States (U.S.). METHODS: Within the U.S. oaks, we tested for phylogenetic and functional trait patterns at different spatial scales, taking advantage of a dated phylogenomic analysis of American oaks and the U.S. Forest Service (USFS) Forest Inventory and Analysis (FIA). KEY RESULTS: We find (1) phylogenetic overdispersion at small grain sizes throughout the U.S. across all spatial extents and (2) a shift from overdispersion to clustering with increasing grain sizes. Leaf traits have evolved in a convergent manner, and these traits are clustered in communities at all spatial scales, except in the far west, where species with contrasting leaf types co-occur. CONCLUSIONS: Our results support the hypotheses that (1) interspecific interactions were important in parallel adaptive radiation of the genus into a range of habitats across the continent and (2) that the diversification process is a critical driver of community assembly. Functional convergence of complementary species from distinct clades adapted to the same local habitats is a likely mechanism that allows distantly related species to coexist. Our findings contribute to an explanation of the long-term maintenance of high oak diversity and the dominance of the oak genus in North America.

Dirofilaria immitis and Angiostrongylus vasorum: The current situation of two major canine heartworms in Portugal.

Cardiopulmonary nematodes are life-threatening pet parasites increasingly reported throughout Europe, with overlapping endemic areas. Dirofilaria immitis is a mosquito-borne whilst Angiostrongylus vasorum is a snail-borne pathogen. Both adult nematodes reside in the pulmonary arteries and right cardiac ventricle of domestic and wild canids, causing a wide spectrum of clinical presentations ranging from cough, dyspnoea and exercise intolerance to severe vascular and pulmonary disease with hearth failure that may lead to death. Information about the prevalence and distribution of cardiopulmonary parasites is essential for the control of animal diseases and, in the case of D. immitis, for the control of potentially associated illnesses in humans. However, in Portugal, heartworm studies are limited to few surveys and case reports, possibly underestimating the relevance of these nematodes. The present work reviews the data on cardiopulmonary dirofilariosis and angiostrongylosis in dogs in Portugal, providing a comprehensive update of the epidemiological situation during the past 20 years.

Pervasive migration across rainforest and sandy coastal plain Aechmea nudicaulis (Bromeliaceae) populations despite contrasting environmental conditions.

Understanding the colonization of extreme marginal habitats and the relative roles of space and environment in maintaining peripheral populations remains challenging. Here, we leverage a system of pairs of rainforest and sandy coastal plain communities that allow us to decouple spatial and environmental effects in the population structure and migration rates of the bromeliad Aechmea nudicaulis. Structure and gene flow between populations were estimated from Bayesian clustering and coalescent-based migration models applied to chloroplast sequence and nuclear microsatellite data. Contrary to our initial expectation, the sharp environmental gradient between rainforest and sandy plains does not seem to have affected the colonization and migration dynamics in A. nudicaulis. Our analyses uncover pervasive gene flow between neighbouring habitats in both chloroplast and nuclear data despite the striking differences in environmental conditions. This result is consistent with a scenario of repeated colonization of the sandy coastal plains from forest populations through seed dispersal, as well as the maintenance of gene flow between habitats through pollination. We also recovered a broad north/south population structure that has been found in other Atlantic rainforest groups and possibly reflects older phylogeographic dynamics.

Balancing selection maintains diversity in a cold tolerance gene in broadly distributed live oaks.

Cold poses major physiological challenges to plants, especially long-lived trees. In trees occurring along variable temperature clines, the expected direction and consequences of selection on cold acclimation ability and freezing tolerance are not straightforward. Here we estimated selection in cold acclimation genes at two evolutionary timescales in all seven species of the American live oaks (Quercus subsection Virentes). Two cold response candidate genes were chosen: ICE1, a key gene in the cold acclimation pathway, and HOS1, which modulates cold response by negatively regulating ICE1. Two housekeeping genes, GAPDB and CHR11, were also analyzed. At the shallow evolutionary timescale, we demonstrate that HOS1 experienced recent balancing selection in the two most broadly distributed species, Q. virginiana and Q. oleoides. At a deeper evolutionary scale, a codon-based model of evolution revealed the signature of negative selection in ICE1. In contrast, three positively selected codons have been identified in HOS1, possibly a signature of the diversification of Virentes into warmer climates from a freezing adapted lineage of oaks. Our findings indicate that evolution has favored diversity in cold tolerance modulation through balancing selection in HOS1 while maintaining core cold acclimation ability, as evidenced by purifying selection in ICE1.