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Horses revolutionized human history with fast mobility1. However, the timeline between their domestication and their widespread integration as a means of transport remains contentious2-4. Here we assemble a collection of 475 ancient horse genomes to assess the period when these animals were first reshaped by human agency in Eurasia. We find that reproductive control of the modern domestic lineage emerged around 2200 BCE, through close-kin mating and shortened generation times. Reproductive control emerged following a severe domestication bottleneck starting no earlier than approximately 2700 BCE, and coincided with a sudden expansion across Eurasia that ultimately resulted in the replacement of nearly every local horse lineage. This expansion marked the rise of widespread horse-based mobility in human history, which refutes the commonly held narrative of large horse herds accompanying the massive migration of steppe peoples across Europe around 3000 BCE and earlier3,5. Finally, we detect significantly shortened generation times at Botai around 3500 BCE, a settlement from central Asia associated with corrals and a subsistence economy centred on horses6,7. This supports local horse husbandry before the rise of modern domestic bloodlines.

This paper reports a high-resolution isotopic study of medieval horse mobility, revealing their origins and in-life mobility both regionally and internationally. The animals were found in an unusual horse cemetery site found within the City of Westminster, London, England. Enamel strontium, oxygen, and carbon isotope analysis of 15 individuals provides information about likely place of birth, diet, and mobility during the first approximately 5 years of life. Results show that at least seven horses originated outside of Britain in relatively cold climates, potentially in Scandinavia or the Western Alps. Ancient DNA sexing data indicate no consistent sex-specific mobility patterning, although three of the five females came from exceptionally highly radiogenic regions. Another female with low mobility is suggested to be a sedentary broodmare. Our results provide direct and unprecedented evidence for a variety of horse movement and trading practices in the Middle Ages and highlight the importance of international trade in securing high-quality horses for medieval London elites.

Introdução: A quantificação de linfócitos T, B e Natural Killer (NK) por citometria de fluxo é o confiável para avaliar a imunocompetência do organismo, crucial para a homeostase e defesa contra patógenos e células danificadas. Alterações nas populações de linfócitos no sangue periférico estão associadas a diversas doenças, incluindo imunodeficiências primárias e secundárias, doenças autoimunes, infecções e câncer. A citometria de fluxo, utilizando anticorpos monoclonais conjugados a fluorocromos, permite a detecção e quantificação dessas subpopulações celulares. Esta tecnologia está em constante avanço, com melhorias contínuas nos equipamentos e procedimentos laboratoriais que aumentam a eficiência e precisão das análises. No entanto, é crucial que os resultados das análises laboratoriais sejam comparáveis independentemente da metodologia e da instrumentação diagnóstica utilizadas. Objetivo/Justificativa: Este estudo teve como objetivo validar e comparar a consistência e a precisão dos resultados da análise de linfócitos T, B e NK utilizando os citômetros de fluxo BD FACSCanto II e BD FACSLyric. Essa validação é fundamental para assegurar que a introdução do novo citômetro BD FACSLyric não comprometa a precisão diagnóstica e o acompanhamento do tratamento clínico, mantendo a qualidade dos diagnósticos realizados. Métodos: Foi realizada uma pesquisa experimental utilizando amostras de sangue periférico de 20 pacientes. As amostras foram marcadas com anticorpos para detecção de linfócitos T, B e NK e suas subpopulações, adquiridas simultaneamente em ambos os citômetros de fluxo. A análise da porcentagem parcial das populações celulares foi realizada utilizando o software Infinicyt. Foram conduzidas análises de correlação e de Bland-Altman. Resultados/Discussão: Todos os coeficientes de correlação de Pearson (r) na Análise de Correlação mostraram-se muito próximos de 1, indicando uma correlação positiva perfeita entre as medições dos dois citômetros de fluxo, com valor de p < 0,001 em todos os casos, o que indica significância estatística. A análise de Bland-Altman revelou pequenas diferenças nas medições entre os equipamentos, demonstrando alta concordância entre as medições dos dois citômetros para a quantificação das células. Os resultados obtidos neste estudo são consistentes com a maioria dos estudos comparativos realizados em outros contextos, evidenciando forte correlação e excelente comparabilidade entre os resultados obtidos na avaliação das subpopulações de linfócitos pelos dois citômetros de fluxo analisados. Conclusão: Este estudo demonstrou que os equipamentos BD FACSCanto II e BD FACSLyric são capazes de fornecer resultados consistentes e comparáveis para a maioria das subpopulações analisadas. Esses achados são cruciais para a validação e padronização dos métodos de análise, especialmente em ambientes clínicos e de pesquisa.

A Leucemia Linfoide Aguda (LLA) é uma neoplasia hematológica caracterizada pela proliferação descontrolada de células progenitoras linfoides B ou T, comprometendo o processo de hematopoese. Representando mais de 80% das leucemias agudas, a LLA de linhagem B é responsável por aproximadamente 85% dos casos, sendo mais comum em crianças. A imunofenotipagem por citometria de fluxo multiparamétrica (CFMP) desempenha um papel crucial na identificação e caracterização das leucemias, permitindo a detecção precisa de marcadores de diferenciação celular. Compreender os perfis de imunofenotipagem em LLA-B pode orientar o desenvolvimento de novas terapias direcionadas. Portanto, o presente estudo teve como objetivo avaliar o perfil de pacientes com LLA-B utilizando a CFMP. A análise de coorte retrospectiva foi realizada por meio da revisão dos prontuários eletrônicos dos pacientes, onde foram coletados dados demográficos e resultados da CFMP no momento do diagnóstico. Este estudo retrospectivo analisou os perfis de imunofenotipagem em 116 pacientes com LLA-B, coletando dados demográficos e de CFMP ao diagnóstico ao longo de cinco anos (jan/2018 a jan/2023). A análise da expressão de CD45 revelou intensidades dim, dim/negativo e negativo em 57, 52 e 6 pacientes, respectivamente. Entre os subtipos de LLA-B, a positividade para CD34 foi associada ao subtipo Comum. Todos os pacientes apresentaram positividade para CD19, e outros marcadores comumente expressos, como CD79a (97,4%), CD22 (81,8%) e CD10 (83,6%), foram positivos na maioria dos casos, ressaltando a importância da identificação alternativa das células B leucêmicas para terapias alvo-específicas. A análise dos subtipos revelou uma prevalência do tipo Comum (75,7%), seguido pelo subtipo Pró-B (14,8%) e Pré-B (9,6%). Observou-se uma predominância do sexo masculino nos subtipos Pró-B e Comum. No subtipo Pré-B, 54,5% dos pacientes eram do sexo feminino, enquanto 45,5% eram do sexo masculino. No subtipo Pró-B, 64,7% dos pacientes eram adultos e 35,3% eram pediátricos. Para o subtipo Comum, 37,9% eram adultos e 62,1% eram crianças. No subtipo Pré-B, associado a uma maior maturação das células leucêmicas, houve uma incidência de 45,5% de adultos e 54,5% de crianças. A variabilidade na expressão de CD45 e a associação de CD34 com o subtipo Comum sublinham a necessidade de uma identificação precisa para o desenvolvimento de estratégias terapêuticas personalizadas. A capacidade da CFMP em detectar e caracterizar de maneira detalhada os marcadores de diferenciação celular oferece uma base sólida para a criação de terapias direcionadas. A interpretação dos resultados reforça a importância clínica da investigação dos marcadores por CFMP, fornecendo informações valiosas para o avanço das estratégias terapêuticas e sugerindo novas direções para pesquisas futuras. A continuidade da pesquisa na área é essencial para aprimorar o tratamento e melhorar os desfechos clínicos para os pacientes com LLA-B.

Acute Myeloid Leukemia (AML) is a severe hematopoietic disorder marked by uncontrolled proliferation of immature myeloid progenitors due to differentiation arrest, with significant immunophenotypic diversity. FLT3-ITD mutations’ prognostic impact varies with NPM1 mutation presence and the FLT3 allelic ratio. While cytogenetic and molecular features are well-established for risk stratification by European LeukemiaNet (2017 ELN), the role of leukemia arrest stages in prognosis remains unclear. This study classified the maturation arrest stages of leukemic blasts in a Brazilian AML cohort, analyzing its association with genetic risk and clinical data. Bone marrow from 148 de novo AML patients (Sep-2015-Jul-2024) was evaluated for FLT3 and NPM1 mutations, FLT3 allelic ratio, and immunophenotyping. MPO cytochemistry stains and phenotypic markers by flow cytometry defined six stages of leukemia arrest (SLA): Hematopoietic Stem Cells-like (HSC-L:CD34+ CD117±CD13-CD33-HLA-DR+MPO-); Multipotent Progenitors-like (MPP-L:CD34+CD117+CD13±CD33±HLA-DR+MPO-); Common Myeloid Progenitors-like (CMP-L:CD34±CD117+CD13+CD33± HLA-DR+MPO+); Granulocyte-Monocyte Progenitors-like (GMP-L:CD34±CD117±CD13±CD33+HLA-DR+MPO+); Monocyte Progenitors-like (MP-L:CD34-CD117±CD13+CD33+HLA-DR+MPO+); and Granulocyte Progenitors-like (GP-L:CD34-CD117±CD13+ CD33+HLA-DR-MPO+). SLA showed highest frequency in MP-L (n = 57, 38.5%) and GP-L (n = 53, 35.8%), followed by GMP-L (n = 23, 15.5%), CMP-L (n = 7, 4.7%), MPP-L (n = 6, 4.1%), and HSC-L (n = 2, 1.4%). Predominance of FLT3wt/NPM1mut patients (n = 80, 54.1%) was observed, with similar distribution among FLT3mut patients (FLT3mut /NPM1mut:n = 32, 21.6%; FLT3mut/NPM1wt:n = 36, 24.3%). CMP-L and GMP-L frequencies were increased in FLT3low/NPM1wt and FLT3high/NPM1wt vs. FLT3wt/NPM1mut (CMP-L:13.33%, 14.28% vs 1.25%, p < 0.05; GMP-L:40%, 42.85% vs 7.5%, p < 0.05). GP-L frequency was decreased in FLT3low/NPM1wt and FLT3 high/NPM1wt vs. FLT3wt/NPM1mut (0%, 9.52% vs 43.75%, p < 0.05). Response to treatment (CR/CRi, CR/CRiMRD+ and PD–Persistence disease) was evaluated after 1st and 2nd cycles of Induction of Remission (1IR, 2IR). GMP-L was increased in PD after 1IR (OR:6.09, 95% CI:1.22–30.19, p < 0.05). Immature (HSC+MPP+CMP) SLA was higher in MRD+ patients compared to MRD- (OR:6.81, 95% CI:0.86–47.05, p < 0.05). Overall survival (p = 0.81) and relapse-free survival (p = 0.27) analyses showed that GMP-L patients relapsed sooner. In 2017 ELN risk assessment, FLT3-ITD stratification was based on allelic ratio and NPM1 mutation status. A high FLT3-ITD allelic ratio with NPM1 mutation indicated intermediate risk, and with wild-type NPM1, it indicated unfavorable risk. The 2022 ELN guidelines classify FLT3-ITD alone as intermediate risk, regardless of NPM1 status. Mutated NPM1 is favorable only when FLT3-ITD is absent. Supporting these findings, FLT3 mutation, regardless of allelic ratio, in the absence of NPM1, led to immature SLA and increased GMP-L frequency. This SLA was linked to PD after the 1IR. Immature SLA significantly impacts prognosis with lower survival rates, higher relapse risks, and increased post-induction remission failure. Our findings enhance the understanding of the cell of origin in AML, and further investigation of the SLA-dependent mechanisms that contribute to leukemia resistance are warranted.

Myelofibrosis (MF), driven by JAK/STAT signaling mutations in hematopoietic stem cells, has the worst survival rates among myeloproliferative neoplasms (MPNs), worsening from pre-fibrotic MF (PFMF) to overt MF (OMF) that often precedes acute myeloid leukemia. Natural Killer (NK) cells have phenotypical alterations that predominate in MF when compared to other MPNs, suggesting that the immune content may contribute to disease burden. We hypothesized that the cytotoxic immune cell profile, along with fibrosis and mutations, drives MF progression. Aiming test this, peripheral blood mononuclear cells (PBMC) from 9 PFMF and 9 OMF, and 9 healthy donors (controls: CT) were analyzed by flow cytometry to profile: NK cell frequency and subtypes (CD45hiCD3-CD19-CD56brightCD16-; CD45hiCD3-CD19-CD56dimCD16+; NK maturation [CD11b-CD27- tolerant (DN), CD27+CD11b- immature secretory (IS), CD27+CD11b+ mature secretory (MS), CD11b+CD27- cytotoxic]; T lymphocytes (CD45hiCD3+) frequency and subtypes (CD4, CD8) and maturation [CD45RO-CD45RA+CD27+ Naïve (N), CD45RO+CD45RA+CD27+ intermediate (INT), CD45RO+CD45RA+CD27- Central Effector (CE), CD45RO+CD45RA-CD27+ Central Memory (CM)]. Immune evasion was assessed by CD62L, and Treg cells (CD4+CD25hiCD127lowFOxp3+) were also quantified. NK cytotoxic capacity was measured by quantifying dead/K562 target cells and NK degranulation (CD107a) after PBMC co-culture. Top of FormBottom of Form Total NK frequency was similar in MF vs CT or OMF vs PFMF. CD56bright (CT 3.4 ± 2.6%, MF 17.7 ± 17.9%, p < 0.05) were increased and CD56dim (CT 82.7 ± 7.6%, MF 53.4 ± 25%, p < 0.05) decreased in MF vs CT. MF showed higher DN (CT 4.5 ± 3.5%, MF 13.9 ± 8.7%, p < 0.05) and IS (CT 0.2 ± 0.3%, MF 1.2 ± 1.2%, p < 0.05), and lower cytotoxic NK cells (CT 92.1 ± 3.8%, MF 77.3 ± 11.2%, p < 0.05). When compared to PFMF, OMF presented higher frequency of cytotoxic NK cells (PFMF 72.2 ± 12%, OMF 82.4 ± 7.9%, p = 0.09). In agreement, reduced NK cytotoxic capacity was observed despite no degranulation changing in MF (death: CT 81.8 ± 4.2%, MF 30.1 ± 7.76%, p < 0.05) and slightly increased cytotoxicity (PFMF 28 ± 6.4%, OMF 31.4 ± 9.9%, p = 0.7) and degranulation (PFMF 10.1 ± 8.8%, OMF 32.17 ± 9.9%, p = 0.1) accompanied fibrosis. No changes in total T lymphocytes or CD4/CD8 frequencies were seen between CT and MF or PFMF and OMF. However, INT and CE reduced in MF (CT 19.8 ± 9.9%, MF 7.3 ± 12.2%, p < 0.05; CT 19.1 ± 16.4%, MF 8.1 ± 13.8%, p < 0.05) and CM decreased in OMF (PFMF 18.9 ± 11.9%, OMF 10.6 ± 9.5%, p < 0.05). CD62L+ CM decreased in MF (CT 21.5 ± 12.8%, MF 6.8 ± 6.8%, p < 0.05) and in OMF (PFMF 9.4 ± 8%, OMF 3.9 ± 3.7%, p = 0.2). Treg frequency decreased in MF (CT 1.9 ± 1.7%, MF 0.25 ± 0.22%, p < 0.05) but increased in OMF (PFMF 0.2 ± 0.13%, OMF 0.3 ± 0.2%, p = 0.5). More advanced MF has a higher degree of fibrosis, which leads to an inflammatory profile. This, in turn, may activate NK cells and result in a more mature but cytotoxic-deficient profile probably due to exhaustion. On the other hand, increased Treg cells found in OMF negatively regulate cytotoxic T cells and its memory compartment, specifically leading to immune evasion via a decrease in CD62L+ CM (associated with better anti-tumor response). The results shed light on the mechanisms involved in MF progression, suggesting that changes in the maturation profile of cytotoxic cells is associated with disease progression, thus inspiring new therapies that target antitumoral cells from the leukemic environment.

Patients with Chronic Myeloid Leukemia (CML) who achieve sustained deep molecular response (DMR) to Tyrosine Kinase Inhibitors (TKIs) are eligible for treatment discontinuation, but only 50% of those achieve long-term treatment-free remission (TFR). Current biomarkers that predict resistance to TKIs as well as loss of the molecular response after TKI interruption have not been defined. We hypothesized that antitumoral immunity mediated by Natural Killer (NK) and T cells may contribute to TFR success and result in distinct responses during treatment and after discontinuation. NK and T cells phenotype were correlated with BCR::ABL1 kinetics and TFR success in a cohort of CML Brazilian patients included in the DES-CML study (Study of treatment discontinuation in CML treated at the Unified Health System), multicenter, prospective, open-label, single-arm, phase 2, non-randomized, ongoing trial, with current data from two Brazilian centers. Using flow cytometry, we evaluated frequency, subtypes, maturation, and receptors expression of NK and T cells from diagnosis to TKI discontinuation. Peripheral blood samples from 20 healthy controls and 68 CML patients were analyzed at different time points: 14 diagnosis (DX), 13 Imatinib failure (F), 7 major molecular response (MMR), 9 DMR, 20 TFR and 5 TFR loss (TFRL). Patients in discontinuation cohort had TKI dose reduced to 50% for 6 months before total suspension. NK cell subtypes were defined as secretory (CD56brightCD16-) or cytotoxic (CD56dimCD16+). We assessed NK maturation markers (CD57 and NKp80), activation (NKG2D,NKp46,DNAM-1), and inhibition receptors (KIR2DL1,TIGIT,NKG2A). PD-1 checkpoint inhibitor was assessed in CD8 T cells. The frequencies of cytotoxic (p < 0.05) and mature (CD57+ and NKp80 p < 0.05) NK cells as well as NKG2D and DNAM-1 (p < 0.05) were lower in the DX, F, and TFRL groups as compared to patients under TKI treatment with DMR, TFR, and healthy individuals. Patients in TFR exhibited a distinct immune profile from TFRL patients that were not influenced by gender, TKI drug, ELTS or SOKAL score. TFR patients presented higher frequency of cytotoxic (p < 0.05) and mature (CD57+ p < 0.05) NK cells, and also NKG2D, DNAM-1,NKp46 (p < 0.05) compared with TFRL. Frequency of PD-1 (p < 0.05) in T cells was decreased in TFR. At the TKI dose reduction timepoint,TFRL patients already had decreased frequency of cytotoxic (p < 0.05) and mature (CD57+ p < 0.05) NK cells as well as of the activating receptor NKG2D (p < 0.05) compared with TFR cases. Conversely, T-cell PD-1 (p < 0.05) expression was higher in TFRL than in TFR patients at the dose reduction point. In conclusion, our study indicates that impaired NK cell function and PD-1 T-cell inhibition are major contributors to molecular responses in CML. Patients with sustained TFR exhibit higher frequencies of cytotoxic, mature and activated NK cells, along with lower PD-1 expression in T cells. These profile contrasts with patients who have persistent disease or lose molecular response post-treatment suspension, who demonstrate impaired NK cell function and increased PD-1 expression. Monitoring parameters such as mature NK content and activating receptors, particularly NKG2D, and PD-1 expression during the de-escalation phase of TKI potentially predicts TFR relapse and can be useful for decision-making in TKI interruption.

Abstract ANTXR1 is one of two cell surface receptors mediating the uptake of the anthrax toxin into cells. Despite substantial research on its role in anthrax poisoning and a proposed function as a collagen receptor, ANTXR1’s physiological functions remain largely undefined. Pathogenic variants in ANTXR1 lead to the rare GAPO syndrome, named for its four primary features: Growth retardation, Alopecia, Pseudoanodontia, and Optic atrophy. The disease is also associated with a complex range of other phenotypes impacting the cardiovascular, skeletal, pulmonary and nervous systems. Aberrant accumulation of extracellular matrix components and fibrosis are considered to be crucial components in the pathogenesis of GAPO syndrome, contributing to the shortened life expectancy of affected individuals. Nonetheless, the specific mechanisms connecting ANTXR1 deficiency to the clinical manifestations of GAPO syndrome are largely unexplored. In this study, we present evidence that ANTXR1 deficiency initiates a senescent phenotype in human fibroblasts, correlating with defects in nuclear architecture and actin dynamics. We provide novel insights into ANTXR1's physiological functions and propose GAPO syndrome to be reconsidered as a progeroid disorder highlighting an unexpected role for an integrin-like extracellular matrix receptor in human aging.

EMILIN1 (elastin-microfibril-interface-located-protein-1) is a structural component of the elastic fiber network and localizes to the interface between the fibrillin microfibril scaffold and the elastin core. How EMILIN1 contributes to connective tissue integrity is not fully understood. Here, we report bi-allelic EMILIN1 loss-of-function variants causative for an entity combining cutis laxa, arterial tortuosity, aneurysm formation, and bone fragility, resembling autosomal-recessive cutis laxa type 1B, due to EFEMP2 (FBLN4) deficiency. In both humans and mice, absence of EMILIN1 impairs EFEMP2 extracellular matrix deposition and LOX activity resulting in impaired elastogenesis, reduced collagen crosslinking, and aberrant growth factor signaling. Collagen fiber ultrastructure and histopathology in EMILIN1- or EFEMP2-deficient skin and aorta corroborate these findings and murine Emilin1-/- femora show abnormal trabecular bone formation and strength. Altogether, EMILIN1 connects elastic fiber network with collagen fibril formation, relevant for both bone and vascular tissue homeostasis.

Abstract Background Escherichia coli is a cost-effective expression system for production of antibody fragments like Fabs. Various yield improvement strategies have been applied, however, Fabs remain challenging to produce. This study aimed to characterize the gene expression response of commonly used E. coli strains BL21(DE3) and HMS174(DE3) to periplasmic Fab expression using RNA sequencing (RNA-seq). Two Fabs, Fabx and FTN2, fused to a post-translational translocation signal sequence, were produced in carbon-limited fed-batch cultivations. Results Production of Fabx impeded cell growth substantially stronger than FTN2 and yields of both Fabs differed considerably. The most noticeable, common changes in Fab-producing cells suggested by our RNA-seq data concern the cell envelope. The Cpx and Psp stress responses, both connected to inner membrane integrity, were activated, presumably by recombinant protein aggregation and impairment of the Sec translocon. The data additionally suggest changes in lipopolysaccharide synthesis, adjustment of membrane permeability, and peptidoglycan maturation and remodeling. Moreover, all Fab-producing strains showed depletion of Mg2+, indicated by activation of the PhoQP two-component signal transduction system during the early stage and sulfur and phosphate starvation during the later stage of the process. Furthermore, our data revealed ribosome stalling, caused by the Fabx amino acid sequence, as a contributor to low Fabx yields. Increased Fabx yields were obtained by a site-specific amino acid exchange replacing the stalling sequence. Contrary to expectations, cell growth was not impacted by presence or removal of the stalling sequence. Considering ribosome rescue is a conserved mechanism, the substantial differences observed in gene expression between BL21(DE3) and HMS174(DE3) in response to ribosome stalling on the recombinant mRNA were surprising. Conclusions Through characterization of the gene expression response to Fab production under industrially relevant cultivation conditions, we identified potential cell engineering targets. Thereby, we hope to enable rational approaches to improve cell fitness and Fab yields. Furthermore, we highlight ribosome stalling caused by the amino acid sequence of the recombinant protein as a possible challenge during recombinant protein production.

Muslims in Italy are an increasingly large and relevant part of the social fabric, although their social condition is still characterized by a “precarious” status. This is explained by a relationship with State institutions that is not yet fully defined in formal terms and by resistance to legitimizing their presence in the public space. In addition, international events have led to the spread of a securitarian political rhetoric and to the intensification of “control” devices on the organized forms and public manifestations of Muslim religiosity. One issue that has concerned political interlocutors has been the training of “imams”. This paper presents the Italian case of training “on” and “of” Islam, analyzing it as a contested field, albeit in a not open and hostile form, between the different social and institutional actors, that is Italian universities, Islamic organizations and transnational Islam and Islam “of the States”. It then analyzes the approach that has been developed and experimented by an Italian State university for the training of imams and murshidat, in collaboration with Italian Islamic organizations and some universities in the Organization of Islamic Cooperation countries, and it also discusses how it fits in as a possible innovative model among the various “assemblages” that have emerged in Europe in recent years.

Domestication of horses fundamentally transformed long-range mobility and warfare1. However, modern domesticated breeds do not descend from the earliest domestic horse lineage associated with archaeological evidence of bridling, milking and corralling2-4 at Botai, Central Asia around 3500 BC3. Other longstanding candidate regions for horse domestication, such as Iberia5 and Anatolia6, have also recently been challenged. Thus, the genetic, geographic and temporal origins of modern domestic horses have remained unknown. Here we pinpoint the Western Eurasian steppes, especially the lower Volga-Don region, as the homeland of modern domestic horses. Furthermore, we map the population changes accompanying domestication from 273 ancient horse genomes. This reveals that modern domestic horses ultimately replaced almost all other local populations as they expanded rapidly across Eurasia from about 2000 BC, synchronously with equestrian material culture, including Sintashta spoke-wheeled chariots. We find that equestrianism involved strong selection for critical locomotor and behavioural adaptations at the GSDMC and ZFPM1 genes. Our results reject the commonly held association7 between horseback riding and the massive expansion of Yamnaya steppe pastoralists into Europe around 3000 BC8,9 driving the spread of Indo-European languages10. This contrasts with the scenario in Asia where Indo-Iranian languages, chariots and horses spread together, following the early second millennium BC Sintashta culture11,12.

This study aims to explore how vulnerable communities, scientists, and artists can collaborate to construct digital community memories to excavate opinion spaces related to socio-natural events, creating a shared and inclusive ground for mutual understanding and collective action. The proposed approach combines web archeology and digital artivism to construct community memories that can be used for social activation in relation to climate change. The research draws on participatory and community-based research techniques, citizen-science approaches, and digital methods to collect dispersed community-memory fragments and create an expressive space for the diverse perspectives and needs of the affected populations. It emphasises the importance of inclusivity and non-polarizing approaches for reconstructing and interpreting narrative streams that propagate in the physical and virtual world, and for representing, through digital art, extreme events and their societal impact. To illustrate the proposed approach, the AquaGranda project is here presented as a case study of community memory related to an extreme event: the high tides that occurred in Venice in November 2019.

Abstract Tagetes dombeyi (Asteraceae, Tageteae), a new species from the Central Andes first collected in the 18th century. A new annual species of Tagetes from the Central Andes of Peru and Bolivia, T. dombeyi, is described and illustrated based on herbarium collections and photographs of the plants in their habitat. Tagetes dombeyi is closely related to Tagetes imbricata, Tagetes multiflora - both of which can occasionally be found mixed with the former on the same herbarium sheet - and Tagetes iltisiana. However, they can be distinguished by diagnostic characters of leaf, corolla, and pappus. A morphological description is provided along with an illustration, a range map, and a key to differentiating the Tagetes species that inhabit Peru and Bolivia.

A Mielofibrose Primária (MFP) é uma neoplasia mieloproliferativa iniciada por mutações na célula-tronco hematopoética (e. g. JAK2V617F) e pode ser classificada de acordo com o grau de fibrose na medula óssea em pré-fibrótica (MFPPF; grau de fibrose

Na Leucemia Mieloide Crônica (LMC), pacientes que alcançam resposta molecular profunda sustentada (RMP) com inibidor de tirosinoquinase (ITQ) podem ter o tratamento descontinuado. A remissão livre de tratamento (RLT) ocorre na minoria dos pacientes e não há biomarcadores preditores de manutenção ou perda de resposta. A imunidade antitumoral mediada por Linfócitos T (LT) e Natural Killer (NK) pode contribuir para manutenção e resultar em respostas distintas durante e após a descontinuação. Correlacionamos o fenótipo T e NK com a cinética de doença residual mensurável (uso de ITQ) e com a perda da RLT (descontinuação-DES). Foram incluídos 10 controles saudáveis (CT) e 23 pacientes com LMC: 8 ao diagnóstico (DX), 4 tratados por mais de 3 anos com RM menor que log 3 (RM4,5) em DES, divididos e acompanhados em meses (m), no qual, 3m (n = 13), 6m (n = 8) e 9m (n = 7), e 8 que recaíram (R) no DES. Até o ponto 6m, os pacientes estavam com 50% da dose do ITQ e, após este ponto, tiveram o ITQ suspenso. Foram avaliados frequência, subtipos e maturação de NK e LT por citometria de fluxo do sangue periférico. Os subtipos NK (CD45hiCD19−CD3−) foram definidos como secretório (CD56brightCD16−) ou citotóxico (CD56dimCD16+), e a maturação funcional das células CD19−CD3−CD56+ foi classificada: CD11b−CD27− (tolerante); CD27+CD11b− (secretória imatura); CD27+CD11b+(secretória madura) e CD11b+CD27− (citotóxica), além dos marcadores de ativação CD57 e NKp80. A frequência destas subpopulações foi avaliada em subgrupos de pacientes de acordo com o nível de RM por RT-qPCR para o gene BCR::ABL1. Em comparação aos CT, amostras de LMC em atividade (DX, RM0,05). Em relação aos subtipos de NK, DX, RM 0,05). Na maturação funcional de NK, DX, RM 0,05). Na ativação de NK, DX, RM4,5 e DES, possuem perfil mais maduro de NK, no qual a atividade citotóxica é semelhante aos CT. Entretanto, pacientes maus respondedores (RM

O monitoramento da Leucemia Mieloide Aguda (LMA) é uma estratégia de avaliação prognóstica para identificar Doença Residual Mensurável (DRM). Indica-se o estudo de DRM por métodos moleculares em casos com mutações NPM1 /translocações tipo core-binding fator; mas a citometria de fluxo multiparamétrica (CFMP) é aplicável a mais de 90% das LMAs e preditora do sucesso terapêutico. Diante da escassez no acesso a métodos moleculares em países em desenvolvimento, um protocolo de determinação de DRM na rotina laboratorial é uma necessidade não atendida. Objetivamos implantar um protocolo de DRM por CFMP para LMA e avaliá-lo quanto à sua eficiência e como fator prognóstico adicional à estratificação de risco clássica. Foram incluídos 191 pacientes com LMA de novo ou secundária à síndrome mielodisplásica, entre 18 e 65 anos (09/2015 a 02/2023), com remissão morfológica completa após tratamento com 1 ou 2 ciclos de indução (ciclo 1:3 dias de daunorrubicina 60 mg/m2 e 7 dias de citarabina 200 mg/m2; ciclo 2:3 dias de daunorrubicina 60 mg/m2 e 6 dias de citarabina 1 g/m2 2x/dia). Na Consolidação foi administrada no ciclo1:citarabina 1 g/m2 2x/dia, 6 dias e no ciclo2: idem 1 ou transplante de Medula Óssea (MO) autólogo com condicionamento BuCy. Os pacientes foram estratificados (ELN2017*-exceto mutações TP53 ou ASXL1) de acordo com cariótipo, testes moleculares e coleta de dados clínicos. As DRMs 1, 2 (Pós-indução) e 4 (Pós consolidação), foram avaliadas com: CD45, CD34, CD117, CD33, CD38, CD19, CD123, CD7, CD56, CD13, CD11b, CD14, CD64, CD300e, CD15, CD133 e HLA-DR. Utilizamos a abordagem de LAIPs e DfN, comparando dados de MO saudável e diagnóstico e sendo DRM+ a frequência de células em CD45+ ≥ 0,1% (mínimo 1 fenótipo anormal). Analisamos Sobrevida Global e Livre de Recaída (SG e SLR) e Incidência Cumulativa de Recaída (ICR) - software R (α = 5%). Não foram encontradas associações entre DRM+ e idade, leucócitos, % de blastos, subtipo morfológico e alterações citogenéticas. Observou-se 21% de DRM+ na DRM1,13% na DRM2 e 12,5 % na DRM4. Na DRM1, a frequência de DRM+ para pacientes de risco baixo, intermediário e alto foi: 46,5%, 33,4%, 20,1% e nas DRMs 2 e 4, 33,4%/ 38,9%/ 27,7% e 48,8%/ 48,8%/ 14,4%, respectivamente. Globalmente as DRMs foram definidas como LAIPs ou DfN em 72,3% e 27,7% das amostras. A expressão de CD56 foi a mais frequente (22%), seguida por alterações em CD34/CD13 (21%) e CD33 (17%). DRMsnegativas foram mais frequentes em pacientes com NPM1mut (DRM1, 69,7%) do que em FLT3-ITD (15,2%). A DRM+ após o 2ºciclo de Indução implicou em maior risco de óbito (HR = 1329 (IC;75,16-235022), p = 0,002), maior ICR (44,5% em 30 meses, p = 0,033) e menor SG (4,4 meses, p = 0,018) no risco intermediário. A detecção de DRM+ não se associou aos riscos favorável e alto. Nossos achados demonstram a padronização da DRM por CFMP em uma população brasileira com tratamento único reprodutível usando painel racional com número limitado de marcadores. A detecção de fenótipos ao diagnóstico (LAIP) é mais útil do que os fenótipos semelhantes a DfN, embora sua combinação tenha validado o tubo proposto como “consenso”, aplicável a todos os casos a despeito do fenótipo inicial. Em pacientes de risco intermediário, o resultado da DRM por CFMP se mostrou fator de risco independente para SG. Sendo este grupo correspondente à cerca 1/3 dos casos, comumente com cariótipo normal e desfechos variáveis, a identificação de fatores prognósticos adicionais e dinâmicos tem grande utilidade e pode guiar as decisões terapêuticas. Por fim, a identificação dos fenótipos de DRM definidos neste protocolo pode contribuir na busca por alvos terapêuticos para erradicação das células leucêmicas resistentes.

A Leucemia Mieloide Aguda (LMA) se origina de anormalidades genéticas ao nível das células-tronco hematopoéticas (CTH), que adquirem propriedades de células-tronco leucêmicas (CTL) com capacidade aumentada de sobrevivência e bloqueio de maturação. Hipotetizamos que a parada de maturação leucêmica por meio de definição fenotípica das células de origem possa acrescentar informações importantes sobre a biologia da LMA e contribuir para a predição de desfechos. Assim, analisamos retrospectivamente em coorte brasileira de pacientes com LMA o perfil fenotípico de maturação de blastos leucêmicos e sua associação com achados clássicos de morfologia, risco genético e informações clínicas. Foram incluídos dados de imunofenotipagem de 359 indivíduos com LMA encaminhados para o estudo ICAML2015, entre 18 e 70 anos, acompanhados durante oito anos, exceto os casos de Leucemia Promielocítica Aguda (LPA) que só foram incluídos após 2020. A expressão de CD34, CD117, CD13, CD33, CD38, (c)MPO e HLA-DR foi avaliada para a categorização nos subtipos HSC: Hematopoietic stem cell; MPP: Multipotent progenitors; CMP: Common myeloid progenitors; GMP: Granulocyte-monocyte progenitors; MP: Monocyte progenitors; GP: Granulocyte progenitors. LMAs tipo HSC foram definidas por CD34+, CD117−/+, HLA-DR+, CD13/33−, MPO−. A expressão de CD13 e 33 definiu o subtipo MPP. A expressão de cMPO entre 10-70% definiu o subtipo CMP e > 70% GMP. Os subtipos MP/GP foram diferenciados pela expressão de HLA-DR no subtipo GP. A reclassificação dos casos demonstrou a seguinte distribuição: HSC (n = 31, 8.64%); MPP (n = 77, 21.45%); CMP (n = 37, 10.31%); GMP (n = 75, 20.89%); MP (n = 50, 13.93%); GP (n = 89, 24.79%). De acordo com a estratificação de risco da European-Leukemia Net 2017, dentre os subtipos HSC, MPP, CMP e GMP predominaram o risco favorável, seguidos de intermediário e adverso. Nos subtipos MP e GP, o risco intermediário foi mais prevalente em relação ao favorável e adverso. A correlação com a classificação morfológica demonstrou que LMA sem maturação, com maturação, mielomonocítica e monocítica se distribuíram de forma equivalente nos subtipos MPP, CMP, GMP, MP e GP; no subtipo HSC não se encontrou leucemia monocítica ou promielocítica, sendo esta caracteristicamente do tipo GP e raramente GMP. Para 325 pacientes, os dados dos genes NPM1, FLT3 -ITD, BAALC, EVI1 e CEBPA estavam disponíveis, sendo encontrada com maior frequência a mutação FLT3 -ITD no subtipo MPP (26/325) e NPM1 em GP (44/325). Em algum ponto de seu seguimento 57/359 pacientes recaíram, representando nos subtipos: HSC (5/31, 16%), MPP (15/77, 19%), CMP (6/37, 16%), GMP (16/75, 21%), MP (6/50, 12%) e GP (9/89, 10%). A sobrevida global foi avaliada em 309/359 pacientes, tendo-se observado maior taxa para o subtipo GP (66%) e menor para CMP (11%) aos 24 meses de seguimento (p = 0.03). Os achados apontam que as correlações entre o estado de parada de maturação da LMA e dados clínicos de estratificação de risco e sobrevida merecem atenção e melhor exploração. É possível que a combinação dos dados genéticos com a classificação imunofenotípica de origem leucêmica seja útil para aferir prognóstico na LMA, pois é plausível que as leucemias mais maduras tenham um microambiente inflamatorio exuberante, capaz de modificar a imunidade antitumoral e contribuir para os desfechos diversos.

A Leucemia Linfoblástica Aguda (LLA) é uma neoplasia de células linfoides imaturas T ou B, sendo a segunda leucemia aguda mais comum no adulto. Acomete cerca de 1 a 5 /100.000 indivíduos por ano nos Estados Unidos e cerca de 66% dos casos são LLA-B. O tratamento da LLA-B consiste em quimio/imunoterapia de alta intensidade, com objetivo primário de restaurar a hematopoese normal. A monitorização de remissão de doença (Doença Residual Mensurável - DRM) é realizada em diferentes etapas do tratamento por citometria de fluxo multiparamétrica (CFMP), sendo a técnica eficaz na detecção de clones leucêmicos pós-tratamento com quimioterapia de alta intensidade. Para pacientes recaídos/refratários a terapia de resgate mais indicada consiste em imunoterapia com agentes específicos, chamados de terapias-alvo (ex: anti-CD3 + anti-CD19 -Blinatumomabe, anti-CD22 -Inotuzumabe + Ozogamicin), e a terapia com células T geneticamente modificadas - CAR-T (do inglês, Chimeric antigen receptor T Cell). Com o surgimento de tais tratamentos surge também a demanda por padronização da monitorização de DRM nos pacientes, visto que terapia pode resultar em perda de expressão de marcadores, o que inviabiliza a análise pelos métodos anteriormente padronizados, enfatizando a possibilidade de recaídas com clones emergentes negativos para a expressão de CD19, por exemplo. Assim, foi objetivo do presente estudo determinar a viabilidade de implantação de novo protocolo para detecção de DRM com base em seleção alternativa de antígenos de células B para pacientes submetidos à terapia convencional ou anti-CD19 por CFMP. Foram analisadas as medulas ósseas de 2 doadores saudáveis, e 2 pacientes LLA-B tratados no Hospital das Clínicas de Ribeirão Preto - FMRP-USP, sendo um submetido à terapia HyperCVAD e 1 paciente refratário, tratado com CAR-T anti-CD19. As amostras foram preparadas de acordo com protocolo Euroflow e avaliadas quanto à expressão dos marcadores (CD45, CD34, CD38, CD81, CD10, CD20, CD19, CD22, CD24, CD66b, CD66c + CD123, CD73) por CFMP em citômetro BD Canto II e analisadas em software Infinicyt por dois observadores diferentes. Os resultados da análise DRM foram expressos como a porcentagem de células nucleadas viáveis que abrigam o fenótipo anormal, sendo a positividade definida quando igual ou maior do que 0,01%. No presente estudo de viabilidade a detecção de células B foi concordante entre os diferentes tubos, independente de marcação de CD19, não havendo diferença nas frequências observadas tanto para amostras de referência como no paciente analisado (p = 0,4478, Kruskal-Wallis). Não foi observada diferença entre as frequências de células anormais detectadas pelos observadores (p = 0,4, Kruskal Wallis) e nossos resultados demonstram reprodutibilidade entre as análises realizadas, com Correlação de Pearson calculada de 0,9995 (IC: 0,9949-0,999, r2 = 0,9989). A análise de Bland-Altman demonstrou não haver divergência entre as metodologias utilizadas. Não houve diferença nas frequências obtidas dos fenótipos caracterizados como DRM+, independente da combinação de marcadores para a gate de células B (CD19xCD22, CD22xCD24, e CD24xCD66b) p = 0,667, Friedman, α=0,05). Em conclusão, nossos dados indicam que o monitoramento da DRM para pacientes com LLA-B tratados com imunoterapia anti-CD19 é possível por CFMP resultando em dados altamente concordantes. O desempenho do resultado preliminar exposto deve garantir resultados comparáveis ao serem avaliados em múltiplos centros, etapa em fase de planejamento, para garantir o avanço da padronização da resposta de DRM por CFMP.

Heterozygosity is a genetic condition in which two or more alleles are found at a genomic locus. Individuals that are the offspring of genetically divergent yet still interfertile parents (e.g. hybrids) are highly heterozygous. One of the most studied aspects in the genomes of these individuals is the loss of their original heterozygosity (LOH) when multi-allelic sites lose one of their two alleles by converting it to the other, or by remaining hemizygous at that site. The region undergoing LOH may involve a single nucleotide polymorphism (SNP) or a longer stretch of DNA. LOH is deeply interconnected with adaptation but the in silico techniques to infer evolutionary relevant LOH blocks are hardly standardised, and a general tool to infer and analyse them across genomic contexts and species is missing. Here, we present JLOH, a computational toolkit for the inference and exploration of LOH blocks in genomes with at least 1% heterozygosity. JLOH only requires commonly available genomic sequencing data as input. Starting from mapped reads, called variants and a reference genome sequence, JLOH infers candidate LOH blocks based on SNP density (SNPs/kbp) and read coverage per position. Considering that most organisms that undergo extensive LOH are hybrids, JLOH has been designed to capture any subgenomic LOH pattern, assigning each LOH block to its subgenome of origin.

Summary: Ancient DNA preserved in the dental pulp offers the opportunity to characterize the genome of some of the deadliest pathogens in human history. However, while DNA capture technologies help, focus sequencing efforts, and therefore, reduce experimental costs, the recovery of ancient pathogen DNA remains challenging. Here, we tracked the kinetics of ancient Yersinia pestis DNA release in solution during a pre-digestion of the dental pulp. We found that most of the ancient Y. pestis DNA is released within 60 min at 37°C in our experimental conditions. We recommend a simple pre-digestion as an economical procedure to obtain extracts enriched in ancient pathogen DNA, as longer digestion times release other types of templates, including host DNA. Combining this procedure with DNA capture, we characterized the genome sequences of 12 ancient Y. pestis bacteria from France dating to the second pandemic outbreaks of the 17th and 18th centuries Common Era.

Summary: Age profiling of archaeological bone assemblages can inform on past animal management practices, but is limited by the fragmentary nature of the fossil record and the lack of universal skeletal markers for age. DNA methylation clocks offer new, albeit challenging, alternatives for estimating the age-at-death of ancient individuals. Here, we take advantage of the availability of a DNA methylation clock based on 31,836 CpG sites and dental age markers in horses to assess age predictions in 84 ancient remains. We evaluate our approach using whole-genome sequencing data and develop a capture assay providing reliable estimates for only a fraction of the cost. We also leverage DNA methylation patterns to assess castration practice in the past. Our work opens for a deeper characterization of past husbandry and ritual practices and holds the potential to reveal age mortality profiles in ancient societies, once extended to human remains.

The extremely rich palaeontological record of the horse family, also known as equids, has provided many examples of macroevolutionary change over the last ~55 Mya. This family is also one of the most documented at the palaeogenomic level, with hundreds of ancient genomes sequenced. While these data have advanced understanding of the domestication history of horses and donkeys, the palaeogenomic record of other equids remains limited. In this study, we have generated genome‐wide data for 25 ancient equid specimens spanning over 44 Ky and spread across Anatolia, the Caucasus, Central Asia and Mongolia. Our dataset includes the genomes from two extinct species, the European wild ass, Equus hydruntinus, and the sussemione Equus ovodovi. We document, for the first time, the presence of sussemiones in Mongolia and their survival around ~3.9 Kya, a finding that should be considered when discussing the timing of the first arrival of the domestic horse in the region. We also identify strong spatial differentiation within the historical ecological range of Asian wild asses, Equus hemionus, and incomplete reproductive isolation in several groups yet considered as different species. Finally, we find common selection signatures at ANTXR2 gene in European, Asian and African wild asses. This locus, which encodes a receptor for bacterial toxins, shows no selection signal in E. ovodovi, but a 5.4‐kb deletion within intron 7. Whether such genetic modifications played any role in the sussemione extinction remains unknown. [ABSTRACT FROM AUTHOR]

Este estudo resulta de construções teóricas e reflexões do conhecimento curricular relacionado à educação, desenvolvidos em uma disciplina de um curso de Pós-graduação. O texto busca refletir sobre as questões curriculares e suas relações com a crise na educação da contemporaneidade, tendo como referência estudiosos do campo de conhecimento curricular e as contribuições da pensadora alemã Hannah Arendt. Ainda, elucida os tipos de currículo e suas respectivas intenções inseridas em seus modelos pedagógicos; métodos didáticos, métodos de pesquisa; concepções de mundo, de conhecimento, de educação, de escola e de avaliação. O percurso metodológico se deu por meio de uma pesquisa bibliográfica em uma abordagem qualitativa. Os resultados apontam que, com a crise da autoridade que se estabeleceu na modernidade atingindo todas as instâncias da sociedade, o campo da educação em seus processos formativos e educacionais viram-se imersos em uma crise educacional que, desde o século XX vem se alastrando mundialmente. Desse modo, justifica-se a necessidade de aprofundamentos teóricos nas questões curriculares do nosso tempo, na intenção de conceber um currículo cada vez mais multicultural considerando de fato a nossa realidade.

Background: Despite recent advances and development of new therapeutic targets and personalized medicine, acute myeloid leukemia (AML) treatment remains defiant. Identification of residual leukemic cells (measurable residual disease -MRD) by Multiparametric Flow Cytometry has been a fast and effective tool, but disease heterogeneity makes MRD assessment a clinical challenge, especially in cases with uncommon features, needing clinical and technical expertise to be accurately performed and interpreted. Aims: To emphasize how absent CD33 expression impacts on interpreting MDR in AML. Methods: We report three clinical cases of AML with absent CD33 assessed for FLT3 and NPM1 mutation, RUNX1/RUNX1T1 and CBFB-MYH11 rearrangements, karyotype and an 8-color panel immunophenotyping (Table 1). Empty gating strategy was used to identify LAIPs (Leukemia Associated-Immunophenotype), different from normal and leukemic stem cells, compared to healthy and regenerating non-AML bone marrow (BM) from diagnosis and follow up (Figures 1 and 2). Assays were performed at the Hematology Laboratory of Ribeirão Preto the Medical School, University of São Paulo, in accordance to Local Ethical Boards. Results: Patient's demographic and clinical data are summarized on Table 2. MFC revealed completely negative CD33 from diagnosis to follow-ups. Treatment consisted of 2 induction cycles (cycle 1: 3 days of Daunorubicin 60 mg/m2 and 7 days of cytarabine 200 mg/m2; cycle 2: 6 days of cytarabine 1 g/m2 twice a day). Consolidation consisted of 1 or 2 cycles of 6 days of cytarabine 1 g/m2 twice a day. BMs were obtained at diagnosis, after 1st and 2nd induction, after 1st and 2nd Consolidation, and 3, 9, 12 and 15 months after consolidation of chemotherapy. Even with different number of follow-ups for each patient, time analysis was preserved. Complete remission (complete or incomplete) was achieved by day 30 after 1st induction and no change in the outcome was reported, even though an altered maturational phenotype persisted with complete absent CD33 expression, during and after treatment, suggesting a very rare polymorphism of CD33 receptor that mimicked MRD positivity. Since CD33 is a common myeloid antigen expressed on malignant blasts in AML, it is prominent evaluate and carefully analyze this marker, once is known that polymorphisms, for example, rs12459419 can affect CD33-antibody conjugated based therapy. CD33low blasts are associated to a more mature AML and its high expression is related to adverse landscapes in pediatric AML, highlighting the importance of CD33 evaluation. Regarding vulnerable steps of MRD assessment, Post –analytical phase must embrace a set of standards in order to prevent report errors. Here we present three cases of AML with absent CD33 from diagnosis till follow-ups visits. CD33 absence mimics positivity for MRD studies. This data emphasizes the importance of caution in MFC analysis and correlation of diagnostic and follow-ups results and interpretation, as constant training of the team.

Este estudo se propõe a apresentar e discutir as tensões e aproximações educacionais encontradas nas obras dos teóricos Hannah Arendt e John Dewey, tendo como ênfase o estudo da categoria experiência. A pesquisa estrutura-se por meio de um levantamento bibliográfico na base de dados Portal de Periódicos CAPES a fim de investigar as concepções de ambos os autores acerca da educação e, ainda, o conceito de experiência concebendo suas relações com o campo da educação e da educação escolar. Assim, o texto busca contribuir para os estudos epistemológicos e com as teorias da educação, de modo a desvelar as contribuições de grandes intelectuais que, como clássicos, se tornaram atemporais para se (re)pensar os desafios educacionais. Identificamos que apesar de configurarem diferentes perspectivas, os escritos de ambos os autores convergem no compromisso de se pensar a educação como um espaço de possibilidades de construção de um pensar autônomo.