Histological interpretation of spitzoid tumours: an extensive machine learning-based concordance analysis for improving decision making.

The histopathological classification of melanocytic tumours with spitzoid features remains a challenging task. We confront the complexities involved in the histological classification of these tumours by proposing machine learning (ML) algorithms that objectively categorise the most relevant features in order of importance. The data set comprises 122 tumours (39 benign, 44 atypical and 39 malignant) from four different countries. BRAF and NRAS mutation status was evaluated in 51. Analysis of variance score was performed to rank 22 clinicopathological variables. The Gaussian naive Bayes algorithm achieved in distinguishing Spitz naevus from malignant spitzoid tumours with an accuracy of 0.95 and kappa score of 0.87, utilising the 12 most important variables. For benign versus non-benign Spitz tumours, the test reached a kappa score of 0.88 using the 13 highest-scored features. Furthermore, for the atypical Spitz tumours (AST) versus Spitz melanoma comparison, the logistic regression algorithm achieved a kappa value of 0.66 and an accuracy rate of 0.85. When the three categories were compared most AST were classified as melanoma, because of the similarities on histological features between the two groups. Our results show promise in supporting the histological classification of these tumours in clinical practice, and provide valuable insight into the use of ML to improve the accuracy and objectivity of this process while minimising interobserver variability. These proposed algorithms represent a potential solution to the lack of a clear threshold for the Spitz/spitzoid tumour classification, and its high accuracy supports its usefulness as a helpful tool to improve diagnostic decision-making.

Making Human Hematopoietic Stem Cells Without Transgenes.

Creating hematopoietic stem cells (HSCs) capable of multilineage engraft while possessing the ability to self-renew stands as a pivotal achievement within the field of regenerative medicine. However, achieving the generation of these cells without transgene expression or teratoma formation has not been fully accomplished. In a recent publication featured in Cell Stem Cell, Piau et al. document the production of functional HSCs derived from human-induced pluripotent stem cells (hiPSCs). They achieved this through a one-step differentiation protocol that notably does not require any transgene expression. hiPSCs-derived HSCs can engraft and self-renew upon serial transplantation and they are able to reconstitute lymphoid, myeloid, and erythroid compartments. This study presents a promising system to further study human HSC ontogeny, and it might represent a crucial step to obtain HSCs.

Young adults with a 22q11.2 microdeletion and the cost of aging with complexity in a population-based context.

PURPOSE: Information about the impact on the adult health care system is limited for complex rare pediatric diseases, despite their increasing collective prevalence that has paralleled advances in clinical care of children. Within a population-based health care context, we examined costs and multimorbidity in adults with an exemplar of contemporary genetic diagnostics. METHODS: We estimated direct health care costs over an 18-year period for adults with molecularly confirmed 22q11.2 microdeletion (cases) and matched controls (total 60,459 person-years of data) by linking the case cohort to health administrative data for the Ontario population (∼15 million people). We used linear regression to compare the relative ratio (RR) of costs and to identify baseline predictors of higher costs. RESULTS: Total adult (age ≥ 18) health care costs were significantly higher for cases compared with population-based (RR 8.5, 95% CI 6.5-11.1) controls, and involved all health care sectors. At study end, when median age was <30 years, case costs were comparable to population-based individuals aged 72 years, likelihood of being within the top 1st percentile of health care costs for the entire (any age) population was significantly greater for cases than controls (odds ratio [OR], for adults 17.90, 95% CI 7.43-43.14), and just 8 (2.19%) cases had a multimorbidity score of zero (vs 1483 (40.63%) controls). The 22q11.2 microdeletion was a significant predictor of higher overall health care costs after adjustment for baseline variables (RR 6.9, 95% CI 4.6-10.5). CONCLUSION: The findings support the possible extension of integrative models of complex care used in pediatrics to adult medicine and the potential value of genetic diagnostics in adult clinical medicine.

Antibodies Reactive to Leptin in Adults with Type 2 Diabetes and Its Relationship with Clinical, Metabolic and Cardiovascular Risk Parameters.

BACKGROUND AND AIMS: Patients with obesity and type 2 diabetes (T2D) have shown alterations in the affinity of IgG anti-leptin antibodies which are possibly related to metabolic alterations. In the present exploratory study, we analyzed serum samples from adults with T2D classified by body mass index (BMI) and evaluated the relationship of IgG anti-leptin antibodies with body composition, metabolic and cardiovascular risk parameters. METHODS: Serum IgG anti-leptin antibodies (total, free and immune complexes fractions) were measured by in-house ELISA. Body composition, metabolic biomarkers (glucose, glycated hemoglobin, lipid profile, insulin, leptin) and cardiometabolic risk indexes (AIP, HOMA-IR, HOMA-ß) were evaluated in one hundred T2D patients. RESULTS: Patients with T2D and obesity presented a decrease in the percentage of IgG anti-leptin immune complexes compared to patients with T2D and overweight (p < 0.0053). Negative correlations of IgG anti-leptin immune complexes with triglycerides (TG) (r=-0.412, p = 0.023) and VLDL-C (r=-0.611, p = 0.017) were found in normal weight T2D patients. Free IgG anti-leptin antibodies correlated positively with TC (r = 0.390, p = 0.032) and LDL-C (r = 0.458, p = 0.011) in overweight individuals with T2D. Finally, total IgG anti-leptin antibodies correlated positively with leptin hormone levels (r = 0.409, p = 0.024) and negatively with HOMA-IR (r =-0.459, p = 0.012) in T2D patients with obesity. CONCLUSIONS: The decrease of IgG anti-leptin immune complexes observed in patients with T2D and obesity suggests a reduction in antibody affinity to the hormone that may impact its transport and signaling, lipid, lipoprotein and insulin metabolism.

Expression of Transcriptional Factors of T Helper Differentiation (T-bet, GATA-3, RORγt, and FOXP3), MIF Receptors (CD44, CD74, CXCR2, 4, 7), and Th1, Th2, and Th17 Cytokines in PBMC from Control Subjects and Rheumatoid Arthritis Patients.

INTRODUCTION: The macrophage migration inhibitory factor (MIF) plays a pivotal role in the development of rheumatoid arthritis (RA). Previous research indicates that MIF can trigger the expression of cytokine profiles associated with Th1, Th2, and Th17 responses in peripheral blood mononuclear cells (PBMC) from both RA patients and control subjects (CS). Despite these, few studies to date precisely elucidate the molecular mechanisms involved. The present study aimed to associate the expression of Th differentiation TF (T-bet, GATA-3, RORγt) with MIF receptors (CD44, CD74, CXCR2, 4, 7) and Th1, Th2, and Th17 cytokines in PBMC from CS and RA patients. METHOD: PBMC from both groups was cultured for 24 h. The expression of the canonical and non-canonical MIF receptors and the TF was determined by flow cytometry. Additionally, multiplex bead analysis was employed to assess the levels of cytokines in the culture supernatants. The findings revealed that T CD4+ lymphocytes in the CS group exhibited a heightened expression of CD74 (p<0.05), whereas RA patients displayed an elevated expression of CXCR7 (p<0.001). Furthermore, T CD4+ lymphocytes from RA patients exhibited greater expression of GATA3, RORγt, and FOXP3, along with elevated levels of pro-inflammatory cytokines compared to the CS group (p<0.001). RESULT: These results indicate that CD74 is more prominently expressed in PBMC from the CS group, whereas CXCR7 is more expressed in PBMC from RA patients. CONCLUSION: We also noted an increased secretion of Th17 profile cytokines in RA, potentially influenced by the activation of FOXP3 via CD74 and RORγt through CXCR7 using the endocytic pathway.

Long COVID-19 and Insulin Autoimmune Syndrome: A Case Report.

PURPOSE: To describe a case report of a patient with symptoms associated with metabolic alterations 1 month after having COVID-19. METHODS: Laboratory tests, clinical evaluations, and body composition assessments were performed by specialists. FINDINGS: The patient presented excessive sweating, hot flashes, dizziness, blurred vision, and seizure. Laboratory tests indicated low glucose levels after convulsions (50, 42.7, and 55 mg/dL), high insulin levels (basal, 638 µIU/mL; 2-hour, >1000 µU/mL), and positivity for anti-insulin antibodies. The patient was diagnosed with insulin autoimmune syndrome. Treatment with azathioprine and nutritional recommendations improved remission. IMPLICATIONS: SARS-CoV-2 infection or vaccination might induce insulin tolerance failure.

The impact of multimorbidity on severe COVID-19 outcomes in community and congregate settings.

Purpose: This study examined the impact of multimorbidity on severe COVID-19 outcomes in community and long-term care (LTC) settings, alone and in interaction with age and sex. Methods: We conducted a retrospective cohort study of all Ontarians who tested positive for COVID-19 between January-2020 and May-2021 with follow-up until June 2021. We used cox regression to evaluate the adjusted impact of multimorbidity, individual characteristics, and interactions on time to hospitalization and death (any cause). Results: 24.5% of the cohort had 2 or more pre-existing conditions. Multimorbidity was associated with 28% to 170% shorter time to hospitalization and death, respectively. However, predictors of hospitalization and death differed for people living in community and LTC. In community, increasing multimorbidity and age predicted shortened time to hospitalization and death. In LTC, we found none of the predictors examined were associated with time to hospitalization, except for increasing age that predicted reduced time to death up to 40.6 times. Sex was a predictor across all settings and outcomes: among male the risk of hospitalization or death was higher shortly after infection (e.g. HR for males at 14 days = 30.3) while among female risk was higher for both outcome in the longer term (e.g. HR for males at 150 days = 0.16). Age and sex modified the impact of multimorbidity in the community. Conclusion: Community-focused public health measures should be targeted and consider sociodemographic and clinical characteristics such as multimorbidity. In LTC settings, further research is needed to identify factors that may contribute to improved outcomes.

Targeting inflammasome-dependent mechanisms as an emerging pharmacological approach for osteoarthritis therapy.

Arthritic diseases have attracted enormous scientific interest because of increased worldwide prevalence and represent a significant socioeconomic burden. Osteoarthritis (OA) is the most prevalent form of arthritis. It is a disorder of the diarthrodial joints, characterized by degeneration and loss of articular cartilage associated with adjacent subchondral bone changes. Chronic and unresolving inflammation has been identified as a critical factor driving joint degeneration and pain in OA. Despite numerous attempts at therapeutic intervention, no effective disease-modifying agents targeting OA inflammation are available to the patients. Inflammasomes are protein complexes known to play a critical role in the inflammatory pathology of several diseases, and their roles in OA pathogenesis have become evident over the last decade. In this sense, it is relevant to evaluate the vital role of inflammasomes as potential modulators of pathogenic features in OA. This review will provide an overview and perspectives on why understanding inflammasome activation is critical for identifying effective OA therapies. We elaborate on the contribution of extracellular mediators from the circulatory system and synovial fluid as well as intracellular activators within the synovial fibroblasts and articular chondrocytes toward invoking the inflammasome in OA. We further discuss the merits of emerging inflammasome targeting therapies and speculate on the potential strategies for inflammasome blockade for OA therapy.

Using Pluripotent Stem Cells to Understand Normal and Leukemic Hematopoietic Development.

Several decades have passed since the generation of the first embryonic stem cell (ESC) lines both in mice and in humans. Since then, stem cell biologists have tried to understand their potential biological and clinical uses for their implementation in regenerative medicine. The hematopoietic field was a pioneer in establishing the potential use for the development of blood cell products and clinical applications; however, early expectations have been truncated by the difficulty in generating bonafide hematopoietic stem cells (HSCs). Despite some progress in understanding the origin of HSCs during embryonic development, the reproduction of this process in vitro is still not possible, but the knowledge acquired in the embryo is slowly being implemented for mouse and human pluripotent stem cells (PSCs). In contrast, ESC-derived hematopoietic cells may recapitulate some leukemic transformation processes when exposed to oncogenic drivers. This would be especially useful to model prenatal leukemia development or other leukemia-predisposing syndromes, which are difficult to study. In this review, we will review the state of the art of the use of PSCs as a model for hematopoietic and leukemia development.

An open-source UFBGA µ-board for wearable devices.

This paper presents the development of an open-source, low-sized, BGA microcontroller breakout board, that can be used for the development of wearable and cyber-physical prototypes. The board is based on the low power, 8-bit, ATtiny20-CCU Microchip AVR microcontroller. The ATtiny20-CCU can be programmed without bootloader, using the Atmel Tiny Programming Interface (TPI), instead of In-System Programming (ISP). The C code used to program the microcontroller can be written and compiled using the Microchip Studio freeware platform. The ATtiny20-CCU Ultra Fine-pitch Ball Grid Array (UFBGA) packaging technology allows the shrinkage of the conceived Electroless Nickel-Immersion Gold (ENIG) Printed Circuit Board (PCB) to a size of only 15.5 × 13 mm. Its low cost also makes it a viable option for developing many educational electronic projects, especially for Instrumentation and Assistive Technology. The contribution of this paper is mainly the hardware prototype design, the PCB manufacturing, building and test of a very low-sized open source µ-breakout PCB board, for wearable Instrumentation applications, towards the emergent Society/Industry 5.0.

Individual variability in space use near power lines by a long-lived territorial raptor.

Evaluating species responses to anthropogenic infrastructures and other habitat changes is often used to assess environmental impacts and to guide conservation actions. However, such studies are generally carried out at the population level, disregarding inter-individual variability. Here, we investigate population- and individual-level responses toward power lines of a territorial raptor, the Bonelli's eagle Aquila fasciata. We used GPS-PTT tracking data of 17 adult eagles to model space use as a function of distance to transmission and distribution lines, while accounting for other habitat features known to affect this species. At population level, eagles increased the intensity of space use in the proximity of power lines (up to 1,000 m), suggesting an attraction effect. At individual level, some eagles shared the general population attraction pattern, while others showed reduced intensity of space use in the proximity of power lines. These differential responses were unrelated to the sex of individuals, but were affected by the characteristics of the power grid, with a tendency for apparent attraction to be associated with individuals occupying home ranges with a denser network of transmission lines and transmission pylons. However, the study could not rule out the operation of other potentially influential factors, such as individual idiosyncrasies, the spatial distribution of prey availability, and the availability of natural perches and nesting sites. Overall, these results suggest that power lines may drive different behaviors and have differential impacts across individuals, with those attracted to the proximity of power lines potentially facing increased risk of mortality through electrocution and collision, and those avoiding power lines being potentially subject to exclusion effects. More generally, our results reinforce the need to understand individual variability when assessing and mitigating impacts of anthropogenic infrastructures.

Función renal en adultos mayores diabéticos e hipertensos

RESUMEN Introducción: La disminución del filtrado glomerular en adultos mayores diabéticos e hipertensos es consecuencia de factores fisiológicos a los que se le añaden cambios vasculares dependientes de alteraciones que producen las enfermedades crónicas subyacentes. Objetivo: Caracterizar la función renal en adultos mayores diabéticos e hipertensos del Policlínico Docente "Ángel Alfonso Ortiz Vázquez", del municipio Manzanillo en la provincia Granma, durante el año 2020. Método: Se realizó un estudio descriptivo, retrospectivo para determinar la función renal en 249 adultos mayores diabéticos e hipertensos pertenecientes al Consultorio Médico de Familia No. 4 del Policlínico Docente "Ángel Alfonso Ortiz Vázquez", del municipio Manzanillo en la provincia Granma, durante el año 2020. Se empleó la fórmula de Cockcroft-Gault y se extrajeron de las historias clínicas las variables: diagnóstico de diabetes mellitus, diagnóstico de hipertensión arterial, edad, sexo, peso y creatinina. Resultados: Predominó el sexo masculino (52,2 %). El rango de edades, comprendido entre 60 y 87 años, tuvo una media de 68,9 ± 7,3. El valor mínimo de filtrado glomerular fue de 27,5 ml/min/1,73 m2, y el máximo fue de 143,7 ml/min/1,73 m2, la media en estos pacientes fue de 69,2 ± 18,7 ml/min/1,73 m2; para los varones fue 71,4 ± 19,0 ml/min/1,73 m2 y de 66,8 ± 18,2 ml/min/1,73 m2 para las mujeres. Predominó la hipertensión arterial (94,4 %) sobre la diabetes mellitus (41,8 %). El filtrado glomerular en los hipertensos (29,7 %) fue inferior a 60 ml/min/1,73 m2. Conclusiones: Un tercio de los pacientes diabéticos e hipertensos del estudio presenta disminución del filtrado glomerular, y esta disminución está asociada al incremento de la edad.

ABSTRACT Introduction: The decrease in glomerular filtration rate in older adults' patients with diabetic and hypertensive problems is a consequence of physiological factors to which are added some vascular changes associated with alterations produced by underlying chronic diseases. Objective: To characterize renal function in older adults' patients with diabetic and hypertensive problems, attended at the Family Doctor's Office No. 4, Policlínico Docente "Ángel Alfonso Ortiz Vázquez", Manzanillo, Granma, in 2020. Methods: A descriptive, retrospective study was carried out in 2020 to determine the renal function in 249 older adults with diabetic and hypertension problems, attended at the Family Doctor's Office No. 4, Policlínico Docente "Ángel Alfonso Ortiz Vázquez", Manzanillo, Granma. The Cockcroft-Gaulty formula was used. The following variables extracted from the medical records were used: diagnosis of diabetes mellitus, diagnosis of hypertension, age, sex, weight and creatinine text. Results: Male sex predominated (52.2 %). The mean age range, between 60 and 87 years, was 68.9 ± 7.3. The minimum glomerular filtration rate was 27.5 ml/min/1.73 m2, and the maximum was 143.7 ml/min/1.73 m2, the mean rate in these patients was 69.2 ± 18.7 ml/min/1.73 m2. For male was 71.4 ± 19.0 ml/min/1.73 m2 and 66.8 ± 18.2 ml/min/1.73 m2 for female. Hypertension predominated (94.4%) over diabetes mellitus (41.8%). The glomerular filtration rate in hypertensive patients (29.7 %) was less than 60 ml/min/1.73 m2. Conclusions: A third of patients with diabetic and hypertension had decreased in glomerular filtration rate, and this decrease was associated with increasing age.

RESUMO Introdução: A diminuição da taxa de filtração glomerular em idosos diabéticos e hipertensos é consequência de fatores fisiológicos aos quais se somam alterações vasculares dependentes de alterações produzidas por doenças crônicas de base. Objetivo: Caracterizar a função renal em idosos diabéticos e hipertensos pertencentes ao Gabinete Médico de Família No. 4 da Policlínico Docente "Ángel Alfonso Ortiz Vázquez", no município de Manzanillo, província de Granma, durante o ano de 2020. Método: Estudo descritivo, retrospectivo, para determinar a função renal em 249 idosos diabéticos e hipertensos pertencentes ao Gabinete Médico de Família No. 4 da Policlínico Docente "Ángel Alfonso Ortiz Vázquez", no município de Manzanillo, província de Granma, durante o ano de 2020. Foi utilizada a fórmula de Cockcroft-Gault e extraídas dos prontuários as variáveis: diagnóstico de diabetes mellitus, diagnóstico de hipertensão arterial, idade, sexo, peso e creatinina. Resultados: Prevaleceu o sexo masculino (52,2%). A faixa etária, entre 60 e 87 anos, teve média de 68,9 ± 7,3. O valor mínimo de filtração glomerular foi 27,5 ml/min/1,73 m2 e o máximo foi 143,7 ml/min/1,73 m2, a média nesses pacientes foi 69,2 ± 18, 7ml/min/1,73 m2; para os homens foi de 71,4 ± 19,0 ml/min/1,73 m2 e 66,8 ± 18,2 ml/min/1,73 m2 para as mulheres. A hipertensão arterial (94,4%) prevaleceu sobre o diabetes mellitus (41,8%). A taxa de filtração glomerular em hipertensos (29,7%) foi inferior a 60 ml/min/1,73 m2. Conclusões: Um terço dos pacientes diabéticos e hipertensos do estudo apresentam diminuição da taxa de filtração glomerular, e essa diminuição está associada ao aumento da idade.

COVID-19 positivity rates, hospitalizations and mortality of adults with and without intellectual and developmental disabilities in Ontario, Canada.

BACKGROUND: Across and within countries there is a need to understand how the COVID-19 pandemic has impacted populations of individuals with intellectual and developmental disabilities (IDD). OBJECTIVE: Rates of COVID-19 positivity for adults with IDD, including Down syndrome, relative to adults without IDD in Ontario, Canada were compared. Health profiles and case-based rates of hospitalizations, intensive care unit admissions, and mortality within 30 days of testing positively were compared for those with IDD, including Down syndrome, versus those without IDD. METHODS: This retrospective cohort study linked health administrative databases using unique encoded identifiers to describe population-level COVID-19 positivity, related hospital use and mortality from January 15, 2020 to January 10, 2021. Incidence rate ratios (RR) and 95% confidence intervals were calculated. RESULTS: Relative to adults without IDD, COVID-19 positivity rates were 1.28 times higher for adults with IDD and 1.42 times higher for adults with Down syndrome. Compared to adults without IDD, adults with IDD were more than twice as likely to be hospitalized following COVID-19 (RR:2.21 (95%CI: 1.93,2.54)) and to die (RR:2.23 (95%CI: 1.86,2.67). These RRs were greater for adults under 65. For adults with Down syndrome, mortality rates were 6.59 (95%CI: 4.51,9.62) times higher than those without IDD. DISCUSSION: In Ontario, Canada, hospitalization and mortality rates associated with COVID-19 are higher for adults with IDD than other adults. These findings should inform vaccination strategies that often prioritize older adults in the general population resulting in people with IDD, who are often in younger age groups, being overlooked.

Características del melanoma cutáneo en dos instituciones de Bogotá, Colombia: Análisis 2012-2016 / Characteristic features of cutaneousmelanoma in two institutions of Bogotá, Colombia: Analysis, 2012-2016

Resumen Introducción: El Melanoma es una neoplasia que se origina de los melanocitos. Este tumor, a pesar de representar solo el 5% de las neoplasias cutáneas, es el responsable del 70% de las muertes producidas por cáncer de piel. En Colombia se ha registrado una supervivencia del 79% a 5 años y en el caso particular del melanoma lentiginoso acral, de tan solo 54%. No obstante, los datos nacionales no son claros, por lo cual es necesario caracterizar a los pacientes con dicho diagnóstico para contribuir con futuros estudios. Metodología: Estudio de corte transversal en el cual se incluyeron y analizaron registros clínicos, demográficos e histopatológicos de pacientes con diagnóstico de melanoma cutáneo atendidos en dos instituciones dermatológicas de Bogotá, Colombia, durante los años 2012-2016. Resultados: Se incluyeron un total de 255 pacientes, la mayoría mujeres (61.0%). Se observó un aumento de 22 a 64 casos diagnosticados en los años 2012 y 2016 respectivamente. El subtipo histológico más frecuente fue el lentigo maligno (33.7%) seguido del melanoma lentiginoso acral (16.1%). La principal localización fue la cara (43.1%). El tiempo entre la detección y la confirmación del diagnóstico por biopsia fue de 17 meses. El 20% de los casos correspondió con un índice de Breslow <1 y el 42.4% de los casos un Clark de I. Conclusiones: Se observó un aumento en el número de casos nuevos de melanoma cutáneo entre los años 2012 y 2016, siendo los subtipos más frecuentemente diagnosticados el lentigo maligno y el melanoma lentiginoso acral. Los tiempos promedio de diagnóstico fueron prolongados.

Abstract Introduction: Melanoma is a cutaneous neoplasm originating from melanocytes. This tumor, despite representing only 5% of skin neoplasms, is responsible for 70% of deaths caused by skin cancer. In Colombia, a mortality of 0.5/100,000 inhabitants has been reported with a survival rate of 79% at 5 years, but in the case of acral lentiginous melanoma, survival is only 54%. However, national data are not clear, which highlights the need to characterize patients with this diagnosis to contribute to further epidemiological studies. Methodology: Cross-sectional study of the clinical, demographic, and histopathological records of patients with diagnosis of cutaneous melanoma, who were attended in two dermatological centers of Bogotá, Colombia, during 2012-2016. Results: A total of 255 patients were included, most of them women (61.0%). An increase from 22 to 64 diagnosed cases was observed from 2012 to 2016, respectively. The most frequent histological subtype was lentigo maligna (33.7%), followed by lentiginous acral melanoma (16.1%). The main location was face (43.1%). The time between detection and confirmation of diagnosis by biopsy was 17 months. 20% of the cases had a Breslow index <1 and 42.4% of the cases were Clark's level I. Conclusions: There was an increase in the number of new cases of cutaneous melanoma between 2012 and 2016, the most frequently diagnosed subtypes being lentigo maligna and acral lentiginous melanoma. The average time of diagnosis was prolonged.

The Novel Role of MIF in the Secretion of IL-25, IL-31, and IL-33 from PBMC of Patients with Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is an autoimmune inflammatory joint disease with complex pathogenesis associated with cytokine dysregulation. Macrophage migration inhibitory factor (MIF) plays a role in systemic inflammation and joint destruction in RA and could be associated with the secretion of other immune-modulatory cytokines such as IL-25, IL-31, and IL-33. For the above, our main aim was to evaluate the IL-25, IL-31, and IL-33 secretion from recombinant human MIF (rhMIF)-stimulated peripheral blood mononuclear cells (PBMC) of RA patients. The rhMIF and lipopolysaccharide (LPS) plus rhMIF stimuli promote the secretion of IL-25, IL-31, and IL-33 (p < 0.05) from PBMC of RA patients. The study groups, the different stimuli, and the interaction between both showed a statistically significant effect on the secretion of IL-25 (p < 0.05) and IL-31 (p < 0.01). The study of the effect of the RA patient treatments and their interaction with the effect of stimuli did not show an interaction between them. In conclusion, our study generates new evidence for the role of MIF in the secretion of IL-25, IL-31, and IL-33 and its immunomodulatory effect on RA.

Dynamic chromatin association of IκBα is regulated by acetylation and cleavage of histone H4.

IκBs exert principal functions as cytoplasmic inhibitors of NF-kB transcription factors. Additional roles for IκB homologues have been described, including chromatin association and transcriptional regulation. Phosphorylated and SUMOylated IκBα (pS-IκBα) binds to histones H2A and H4 in the stem cell and progenitor cell compartment of skin and intestine, but the mechanisms controlling its recruitment to chromatin are largely unknown. Here, we show that serine 32-36 phosphorylation of IκBα favors its binding to nucleosomes and demonstrate that p-IκBα association with H4 depends on the acetylation of specific H4 lysine residues. The N-terminal tail of H4 is removed during intestinal cell differentiation by proteolytic cleavage by trypsin or chymotrypsin at residues 17-19, which reduces p-IκBα binding. Inhibition of trypsin and chymotrypsin activity in HT29 cells increases p-IκBα chromatin binding but, paradoxically, impaired goblet cell differentiation, comparable to IκBα deletion. Taken together, our results indicate that dynamic binding of IκBα to chromatin is a requirement for intestinal cell differentiation and provide a molecular basis for the understanding of the restricted nuclear distribution of p-IκBα in specific stem cell compartments.

Downregulation of Inflammatory Cytokine Release from IL-1ß and LPS-Stimulated PBMC Orchestrated by ST2825, a MyD88 Dimerisation Inhibitor.

The inflammatory process implicates homeostasis disruption and increased production of inflammatory mediators. Myeloid differentiation primary response 88 (MyD88) is an essential protein recruited after lipopolysaccharide (LPS) and interleukin (IL)-1ß stimulation, a process that converges in nuclear factor kappa B (NF-κB) activation, as well as a transcription of several genes of both pro- and anti-inflammatory cytokines. The inhibition of MyD88 has shown efficacy by decrease inflammatory response, and has demonstrated potential application as a therapeutic target in chronic diseases. In this study, we investigate the effect of MyD88 dimerisation inhibitor ST2825 on cytokine production from rhIL-1ß and LPS-stimulated peripheral blood mononuclear cells (PBMC) from healthy blood donors (HBD). ST2825 significantly downregulates the production of IFN-γ, IL-6, IL-12, IL-2, IL-15, IL-7, VEGF, IL-1Ra, IL-4, IL-5, IL-13 and IL-9 (p < 0.05) in LPS-stimulated PBMC. Moreover, ST2825 had a relatively low impact on IL-1ß signalling pathway inhibition, showing that only a few specific cytokines, such as IFN-γ and IL-1Ra, are inhibited in rhIL-1ß-stimulated PBMC (p < 0.01). In conclusion, MyD88 dimerisation inhibitor ST2825 showed high efficacy by inhibiting pro- and anti-inflammatory cytokine production in LPS-stimulated PBMC. Moreover, although rhIL-1ß induced a sustained cytokine production (p < 0.05), ST2825 did not show a significant effect in the secretion of neither pro- nor anti-inflammatory cytokines in rhIL-1ß-stimulated PBMC.

Using an apex predator for large-scale monitoring of trace element contamination: Associations with environmental, anthropogenic and dietary proxies.

Understanding the levels and drivers of contamination in top predators is important for their conservation and eventual use as sentinels in environmental monitoring. Therefore, metals and trace elements were analyzed in feathers of Bonelli's eagles (Aquila fasciata) from southern Portugal in 2007-2013, where they are believed to be exposed to a wide range of contamination sources such as agricultural land uses, urban areas, active and abandoned mines and a coal-fired power plant. We focused on concentrations of aluminum (Al), arsenic (As), copper (Cu), chromium (Cr), mercury (Hg), lead (Pb), selenium (Se) and zinc (Zn), as these contaminants are potentially associated with those sources and are known to pose a risk for terrestrial vertebrates. Stable isotope values of nitrogen (δ15N: 15N/14N), carbon (δ13C: 13C/12C) and sulphur (δ34S: 34S/32S) were used as dietary proxies to control for potential effects of prey composition on the contamination pattern. The spatial distribution of potential contamination sources was quantified using geographic information systems. Concentrations of Hg in the southern part of the study area were above a reported toxicity threshold for raptors, particularly in territories closer to a coal-fired power plant at Sines, showing that contamination persisted after a previous assessment conducted in the 1990s. Hg and Se levels were positively correlated with δ15N, which indicates biomagnification. Concentrations of As, Cr, Cu, Pb and Zn were generally low and unrelated to mining- or industrial activities, indicating low environmental background concentrations. Al was found at higher concentrations in the southernmost areas of Portugal, but this pattern might be related to external soil contamination on feathers. Overall, this study indicates that, among all elements studied, Hg seems to be the most important contaminant for Bonelli's eagles in southern Portugal, likely due to the power plant emissions and biomagnification of Hg in terrestrial food webs.

Unravelling population processes over the Late Pleistocene driving contemporary genetic divergence in Palearctic buzzards.

Population range expansions and contractions as a response to climate and habitat change throughout the Quaternary are known to have contributed to complex phylogenetic and population genetic events. Speciation patterns and processes in Palearctic buzzards (genus Buteo) are a long-standing example of morphological and genetic data incongruence, attributed to panmixia, habitat range shifts, contact zones, and climate change. Here we assess the systematics, phylogeography and population genetic structure of three nominal species of Palearctic buzzards, Buteo buteo (including B. b. vulpinus), B. rufinus (including B. r. cirtensis) and B. hemilasius. Phylogenetic analyses inferred from mitochondrial data recover B. hemilasius as sister to the sister clades B. r. rufinus and B. buteo complex (B. b. buteo, B. b. vulpinus, but also including B. r. cirtensis). In contrast, we find an unresolved genetic delimitation inferred from four nuclear loci, suggesting an ancestral genetic pool for all species. Time-trees suggest population contractions and expansions throughout the Pleistocene, which likely reflect habitat change and contrasting ecological niche requirements between species. Microsatellite-based extended Bayesian skyline plots reveal relatively constant population sizes for B. hemilasius, B. r. rufinus, and B. b. vulpinus, in contrast to a dramatic population expansion in B. r. cirtensis within the last 3 kya. Overall, our study illustrates how complex population processes over the Late Pleistocene have shaped the patterns of genetic divergence in Palearctic buzzards, due to the joint effects of shared ancestral polymorphisms, population expansions and contractions, with hybridization at contact zones leading to admixture and introgression.

Macrophage migration inhibitory factor polymorphisms are a potential susceptibility marker in systemic sclerosis from southern Mexican population: association with MIF mRNA expression and cytokine profile.

INTRODUCTION: Systemic sclerosis (SSc) is a complex autoimmune disease, characterized by microvascular lesions, autoimmunity, and fibrosis. It is suggested that MIF participates in the amplification of the proinflammatory process in SSc; moreover, the promoter polymorphisms - 794 CATT5-8 (rs5844572) and - 173G>C (rs755622) in the MIF gene have been associated with an increase in MIF serum levels in several autoimmune diseases. The aim of this study was to analyze the relationship of the - 794 CATT5-8 and - 173G>C MIF polymorphisms with mRNA expression, MIF serum levels, and the Th1/Th2/Th17 cytokine profile in SSc. MATERIALS AND METHODS: A case-control study was carried out that included 50 patients with SSc and 100 control subjects (CS). Both polymorphisms were genotyped by PCR and PCR-RFLP. MIF levels were measured by ELISA kit. The cytokine profile and the MIF mRNA expression were quantified by BioPlex MagPix system and real-time PCR, respectively. RESULTS: An association between the - 794 CATT7 and - 173*C MIF alleles and the 7C haplotype with SSc susceptibility was found (p < 0.05). Also, the 7C haplotype was associated with increased MIF mRNA expression (p = 0.03) in SSc. In addition, an increase of IL-1ß and IL-6 serum levels in SSc patients was found as well as a positive correlation between MIF serum levels and Th1 and Th17 cytokine profiles. CONCLUSION: The MIF 7C haplotype is a susceptibility marker for SSc in the southern Mexican population and is associated with MIF mRNA expression. Moreover, there is a positive correlation between MIF serum levels and Th1 and Th17 inflammatory response in SSc.