A standardized approach to evaluation and reporting of synovial histopathology in two surgically induced murine models of osteoarthritis.

OBJECTIVE: Synovial pathology has been linked to osteoarthritis (OA) pain in patients. Microscopic grading systems for synovial changes in human OA have been described, but a standardized approach for murine models of OA is needed. We sought to develop a reproducible approach and set of minimum recommendations for reporting of synovial histopathology in mouse models of OA. METHODS: Coronal and sagittal sections from male mouse knee joints subjected to destabilization of medial meniscus (DMM) or partial meniscectomy (PMX) were collected as part of other studies. Stains included Hematoxylin and Eosin (H&E), Toluidine Blue (T-Blue), and Safranin O/Fast Green (Saf-O). Four blinded readers graded pathological features (hyperplasia, cellularity, and fibrosis) at specific anatomic locations. Inter-reader agreement of each feature score was determined. RESULTS: There was acceptable to very good agreement when using 3-4 individual readers. After DMM and PMX, expected medial predominant changes in hyperplasia and cellularity were observed, with fibrosis noted at 12 weeks post-PMX. Synovial changes were consistent from section to section in the mid-joint area. When comparing stains, H&E and T-blue resulted in better agreement compared to Saf-O stain. CONCLUSIONS: To account for the pathologic and anatomic variability in synovial pathology and allow for a more standardized evaluation that can be compared across studies, we recommend evaluating a minimum set of 3 pathological features at standardized anatomic areas. Further, we suggest reporting individual feature scores separately before relying on a single summed "synovitis" score. H&E or T-blue are preferred, inter-reader agreement for each feature should be considered.

T-Cell Immunoglobulin and Mucin Domain 3 (TIM-3) Gene Expression as a Negative Biomarker of B-Cell Acute Lymphoblastic Leukemia.

B-cell acute lymphoblastic leukemia (B-ALL) accounts for 85% of all childhood ALL. Malignancies exhaust T and B cells, resulting in an increased expression of immune checkpoint receptors (ICRs), such as T-cell immunoglobulin and mucin domain 3 (TIM-3). TIM-3 has been found to be dysregulated in different types of cancer. However, there is a lack of rigorous studies on the TIM-3 expression in B-ALL. The current study aimed to measure the expression of TIM-3 at the gene and protein levels and evaluate the potential of TIM-3 as a biomarker in B-ALL. A total of 28 subjects were recruited between 2021 and 2023, comprising 18 subjects diagnosed with B-ALL and 10 non-malignant healthy controls. The B-ALL patients were divided into three groups: newly diagnosed (four patients), in remission (nine patients), and relapse/refractory (five patients). The expression levels of TIM-3 were evaluated using the real-time qPCR and ELISA techniques. The results revealed that the TIM-3 expression was significantly downregulated in the malignant B-ALL patients compared to the non-malignant healthy controls in the mRNA (FC = -1.058 ± 0.3548, p = 0.0061) and protein blood serum (p = 0.0498) levels. A significant TIM-3 gene reduction was observed in the relapse/refractory cases (FC = -1.355 ± 0.4686, p = 0.0327). TIM-3 gene expression allowed for significant differentiation between patients with malignant B-ALL and non-malignant healthy controls, with an area under the curve (AUC) of 0.706. The current study addressed the potential of reduced levels of TIM-3 as a negative biomarker for B-ALL patients.

The sensitivities and adaptive capacity of public lands visitors.

Lands and waters administered by governing entities for public use (i.e., "public lands") are subject to changing social and ecological conditions (e.g., overcrowding, drought). Public lands managers are often tasked with addressing these changes while balancing conservation goals and public use mandates, and their decisions can significantly and inequitably impact visitor sensitivities to different types of exposures. To gain insights into visitor sensitivities and their adaptive capacity to mitigate the impacts of exposures, we draw upon a comprehensive monitoring effort conducted in collaboration with the U.S. Fish and Wildlife Service (Service) to understand visitor experiences on national wildlife refuges (refuges). We collected data from 10,556 visitors to 68 refuges during 2018-2019, then segmented respondents into unique visitor types based on their frequency of visiting "this refuge" where they were contacted, their participation across different activities at that refuge, and visits to other public lands for purposes of their primary activity, all during the 12 months prior to being contacted. We then explored differences among the resulting visitor types in their (a) purpose of visit, (b) satisfaction with opportunities during their visit, and (c) demographic characteristics. Finally, we used external data sources to explore the sensitivities and adaptive capacity of visitors' home communities. Our approach identified eight types of visitors with distinct sensitivities and adaptive capacities. For example, the type categorized as "most sensitive" due to activity specialization and site dependency was more likely to engage in activities (e.g., fishing, hunting) that may be subsistence uses of public lands and more often lived in communities with reduced adaptive capacity. Our assessment supports public lands decision-making by helping to understand and address social inequities that may arise or be exacerbated by rapidly changing conditions.

Sexual dimorphism of the synovial transcriptome underpins greater PTOA disease severity in male mice following joint injury.

OBJECTIVE: Osteoarthritis (OA) is a disease with sex-dependent prevalence and severity in both human and animal models. We sought to elucidate sex differences in synovitis, mechanical sensitization, structural damage, bone remodeling, and the synovial transcriptome in the anterior cruciate ligament rupture (ACLR) mouse model of post-traumatic OA (PTOA). DESIGN: Male and female 12-week-old C57/BL6J mice were randomized to Sham or noninvasive ACLR with harvests at 7d or 28d post-ACLR (n = 9 per sex in each group - Sham, 7d ACLR, 28d ACLR). Knee hyperalgesia, mechanical allodynia, and intra-articular matrix metalloproteinase (MMP) activity (via intravital imaging) were measured longitudinally. Trabecular and subchondral bone (SCB) remodeling and osteophyte formation were assessed by µCT. Histological scoring of PTOA, synovitis, and anti-MMP13 immunostaining were performed. NaV1.8-Cre;tdTomato mice were used to document localization and sprouting of nociceptors. Bulk RNA-seq of synovium in Sham, 7d, and 28d post-ACLR, and contralateral joints (n = 6 per group per sex) assessed injury-induced and sex-dependent gene expression. RESULTS: Male mice exhibited more severe joint damage at 7d and 28d and more severe synovitis at 28d, accompanied by 19% greater MMP activity, 8% lower knee hyperalgesia threshold, and 43% lower hindpaw withdrawal threshold in injured limbs compared to female injured limbs. Females had injury-induced catabolic responses in trabecular and SCB, whereas males exhibited 133% greater normalized osteophyte volume relative to females and sclerotic remodeling of trabecular and SCB. NaV1.8+ nociceptor sprouting in SCB and medial synovium was induced by injury and comparable between sexes. RNA-seq of synovium demonstrated similar injury-induced transcriptomic programs between the sexes at 7d, but only female mice exhibited a transcriptomic signature indicative of synovial inflammatory resolution by 28d, whereas males had persistent pro-inflammatory, pro-fibrotic, pro-neurogenic, and pro-angiogenic gene expression. CONCLUSION: Male mice exhibited more severe overall joint damage and pain behavior after ACLR, which was associated with persistent activation of synovial inflammatory, fibrotic, and neuroangiogenic processes, implicating persistent synovitis in driving sex differences in murine PTOA.

Epidermal growth factor deficiency predisposes to progressive renal disease.

Epidermal growth factor (EGF) is produced in the kidney by thick ascending limbs of the loop of Henle and by distal convoluted tubules (DCTs). Reduced urinary EGF levels have been associated with chronic kidney disease but it is not known whether physiological levels of EGF protect the kidney from progressive renal disease. Here, we show that EGF-deficient mice on a mixed genetic background had increased urinary microalbumin, and a subset of these mice developed severe progressive renal disease with azotemia that was not seen in WT or TGFα-deficient littermates with this mixed genetic background. These azotemic EGF-deficient mice developed crescentic glomerulonephritis linked to HB-EGF/EGFR hyperactivation in glomeruli, as well as attenuation of the proximal tubule brush border, distal convoluted tubule (DCT) dilatation, and kidney fibrosis associated with renal ß-catenin/mTOR hyperactivation. The observation of these severe renal pathologies only in a subset of EGF-deficient mice suggests that independent segregation of strain-specific modifier alleles contributes to the severity of the renal abnormalities that only manifest when EGF is lacking. These findings link the lack of EGF to renal pathologies in the adult mammalian kidney, in support of a role of physiological levels of EGF for maintaining the function of glomeruli, proximal tubules, and DCTs. These observations suggest that diminished EGF levels predispose kidneys to progressive renal disease.

Golgi signalling proteins GOLPH3, MYO18A, PITPNC1 and RAB1B: implications in prognosis and survival outcomes of AML patients.

BACKGROUND: The role of different Golgi signalling proteins remains unexplored in the progression and spread of acute myeloid leukaemia (AML), whom all interact together in a way that facilitates proliferation and differentiation of myeloid lineage cells. OBJECTIVE: Since Golgi apparatus acts as master brain in membrane trafficking and signalling events that affect cell polarity necessary for migration, division, or differentiation; this study aims to explore the association between signalling proteins and the diagnosis, prognosis, and survival of AML patients. MATERIAL AND METHODS: This study comprised 70 newly diagnosed AML patients and 20 healthy controls to investigate the serum levels of signalling proteins; Golgi Phosphoprotein 3 (GOLPH3), Myosin 18A (MYO18A), Cytoplasmic Phosphatidylinositol Transfer Protein 1 (PITPNC1) and Ras-Associated Binding Protein 1B (RAB1B). RESULTS: AML patients showed higher serum levels of GOLPH3, MYO18A, PITPNC1 and RAB1B when compared to control (p < 0.001). A significant negative correlation was found between the patients' overall survival and GOLPH3 (p = 0.001), MYO18A (p = 0.011), PITPNC1 (p = 0.001) and RAB1B (p = 0.042). Results were confirmed by Kaplen-Meier survival analysis showing lower survival estimates in patients with higher GOLPH3 (p = 0.014), MYO18A (p = 0.047), PITPNC1 (p = 0.008) and RAB1B (p = 0.033) serum levels. CONCLUSION: GOLPH3, MYO18A, PITPNC1 and RAB1B maybe promising diagnostic and prognostic biomarkers in AML patients.

Effect of semantics in the study of tolerance for wolves.

As conservation scholars increasingly recognize the critical role of human thought and behavior in determining the persistence of biodiversity across the globe, a growing line of inquiry regarding the validity and comparability of previous applications of core psychological concepts has emerged. Specifically, inconsistent measurement and use of terms, such as attitudes and acceptance, reveal important questions about previous approaches. Given that these concepts differ by definition, yet have been used interchangeably, we explored what drives differences in people's responses when each concept is operationalized in the context of a contested wildlife species, the gray wolf (Canis lupus). To do so, we used data from a 2014 survey of U.S. residents (n = 1287) to test how measures of six concepts (i.e., acceptance, attitudes, benefits, risks, [prior] behavior, and behavioral intentions) often employed in the conservation social sciences were related with a broad set of possible explanatory variables. Despite moderate to strong correlations between all concepts measured (| Pearson's r | = 0.39-0.65, p < 0.001), results revealed considerable variation in their respective relationships with identical explanatory variables. Specifically, although wildlife value orientation (i.e., domination or mutualism) operated fairly consistently across cognitive and behavioral concepts, the relationship between the six concepts and other factors, such as social trust, identification with various interest groups (i.e., hunter, farmer, or rancher, environmentalist, and animal rights advocate), and political ideology (i.e., liberal vs. conservative), varied considerably. Our findings underscore that differences exist in these measures, which could have serious implications for conservationists integrating social science findings in their decision-making processes if they are unaware of the theoretical underpinnings of and distinctions between core psychological concepts.

Efectos de la semántica en los estudios de tolerancia a los lobos Resumen Los académicos dedicados a la conservación reconocen cada vez más lo importantes que son el pensamiento y el comportamiento humano para definir la persistencia de la biodiversidad a nivel mundial, por lo que ha emergido una creciente línea de indagación con respecto a la validez y la comparabilidad de las aplicaciones previas de conceptos psicológicos fundamentales. Más específicamente, las medidas incompatibles y el uso de términos como actitudes y aceptación revelan preguntas importantes sobre las estrategias anteriores. Ya que estos conceptos difieren por definición y aun así se han usado indistintamente, decidimos explorar qué impulsa las diferencias en las respuestas de las personas cuando cada concepto opera en el contexto de una especie de fauna controvertida: el lobo gris (Canis lupus). Para lograr esto, usamos datos de un censo de 2014 aplicado a residentes estadunidenses (n = 1,287) para analizar cómo la medida de seis conceptos usados frecuentemente en las ciencias sociales de la conservación (aceptación, actitudes, beneficios, riesgos comportamiento [previo] e intenciones conductuales) se relacionan con un amplio conjunto de variables explicativas posibles. A pesar de las correlaciones moderadas y fuertes entre todos los conceptos medidos (| Pearson's r | = 0.39 a 0.65, p < 0.001), los resultados revelaron una variación considerable en sus respectivas relaciones con las variables explicativas idénticas. De manera más precisa, aunque la orientación del valor de la fauna (es decir, dominancia y mutualismo) operó uniformemente en los conceptos cognitivos y conductuales, la relación entre los seis conceptos y otros factores, como la confianza social, identificación con varios grupos de interés (cazador, agricultor o ranchero, ambientalista, defensor de los derechos animales) e ideología política (liberal vs conservador) variaron considerablemente. Nuestros resultados destacan las diferencias que existen en estas medidas, las cuales podrían tener repercusiones serias para los conservacionistas que integran los resultados de las ciencias sociales dentro de sus procesos de toma de decisiones si no están concientes de las teorías fundamentales y las distinciones entre los conceptos psicológicos fundamentales.

Successful Management of Severe Hyperhaemolysis with Combined Tocilizumab and Rituximab in Non-Transfusion-Dependent Thalassaemia: A Case Report.

Introduction: This is the fourth case reporting the administration of tocilizumab to control hyperhaemolysis. It was administered with rituximab to stop hyperhaemolysis refractory to frontline therapy. Hyperhaemolysis is a rare life-threatening subtype of delayed haemolytic transfusion reaction. Refractory cases pose a clinical challenge with no standard of care to date. Case Presentation: A 29-year-old lady with non-transfusion-dependent thalassaemia presented with refractory hyperhaemolysis necessitating the administration of rituximab. This was complicated with anaemic heart failure and altered sensorium exacerbated with further transfusions. A nadir haemoglobin of 2.1 g/dL was reached after the initiation of rituximab, and her condition was too critical to wait for the slow expected improvement. Hence, tocilizumab was given as a bridging therapy to block haemolysis till the delayed onset of radical treatment. Conclusion: Tocilizumab can be effectively combined with rituximab to stop hyperhaemolytic episode refractory to first-line treatment when a prompt response is needed.

An expansion of phenotype: novel homozygous variant in the MED17 identified in patients with progressive microcephaly and global developmental delay.

Global developmental delay (GDD) is a lifelong disability that affects 1-3% of the population around the globe. It is phenotypically variable and highly heterogeneous in terms of the underlying genetics. Patients with GDD are intellectually disabled (ID) manifesting cognitive impairment and deficient adaptive behavior. Here, we investigated a two-looped consanguineous family segregating severe ID, seizure, and progressive microcephaly. Magnetic resonance imaging (MRI) of the brain showed mild brain atrophy and myelination defect. Whole exome sequencing (WES) was performed on the DNA samples of two patients and a novel homozygous missense variant (Chr11:g0.93528085; NM_004268.5_c.871T > C; p. Trp291Gly) was identified in the MED17 gene. Sanger sequencing revealed that the identified variant is heterozygous in both parents and healthy siblings. This variant is conserved among different species, causes a non-conserved amino acid change, and is predicted deleterious by various in silico tools. The variant is not reported in population variant databases. MED17 (OMIM: 613668) encodes for the mediator of RNA polymerase II transcription complex subunit 17. Structure modeling of MED17 protein revealed that Trp291 is involved in different inter-helical interactions, providing structural stability. Replacement of Trp291Gly, a less hydrophobic amino acid loses the inter-helical interaction leading to a perturb variant of MED17 protein.

Serum levels of FAK and some of its effectors in adult AML: correlation with prognostic factors and survival.

Focal adhesion kinase (FAK), human myofibrillogenesis regulator-1 (MR-1), ephrin receptor type A4 (EphA4), proto-oncogene tyrosine kinase Src (Src), and protein kinase C (PKC) are important markers in proliferation, survival, and migration in some cancers. However, the significance of each is still unclear in different malignancies, including acute myeloid leukemia (AML). Therefore, this study was conducted to investigate their serum levels in Egyptian adult de novo AML patients (n = 70) against healthy volunteers (n = 20). We managed to study the correlation between each pair and to investigate their association with diagnosis, prognosis, and survival. Serum levels were analyzed using enzyme-linked immunosorbent assay (ELISA). We found that FAK, MR-1, Src, and PKC serum levels were significantly higher in AML patients compared to control (p < 0.0001), and this was associated with significantly lower EphA4 level (p < 0.0001). Interestingly, we also observed a significant negative correlation of FAK (p = 0.027), MR-1 (p = 0.003), Src (p = 0.038), and PKC (p = 0.03) with patients' overall survival (OS) while there was a positive significant correlation between EphA4 and OS (p = 0.007). In conclusion, this study suggests that FAK, MR-1, EphA4, Src, and PKC may be used as early diagnostic and prognostic markers with high sensitivity and specificity in AML patients and thus may be incorporated into the patients' early diagnostic and prognostic panels.

Automated algorithm for calculation of setup corrections and planning target volume margins for offline image-guided radiotherapy protocols.

PURPOSE: Each radiotherapy center should have a site-specific planning target volume (PTV) margins and image-guided (IG) radiotherapy (IGRT) correction protocols to compensate for the geometric errors that can occur during treatment. This study developed an automated algorithm for the calculation and evaluation of these parameters from cone beam computed tomography (CBCT)-based IG-intensity modulated radiotherapy (IG-IMRT) treatment. METHODS AND MATERIALS: A MATLAB algorithm was developed to extract the setup errors in three translational directions (x, y, and z) from the data logged by the CBCT system during treatment delivery. The algorithm also calculates the resulted population setup error and PTV margin based on the van Herk margin recipe and subsequently estimates their respective values for no action level (NAL) and extended no action level (eNAL) offline correction protocols. The algorithm was tested on 25 head and neck cancer (HNC) patients treated using IG-IMRT. RESULTS: The algorithms calculated that the HNC patients require a PTV margin of 3.1, 2.7, and 3.2 mm in the x-, y-, and z-direction, respectively, without IGRT. The margin can be reduced to 2.0, 2.2, and 3.0 mm in the x-, y-, and z-direction, respectively, with NAL and 1.6, 1.7, and 2.2 mm in the x-, y-, and z-direction, respectively, with eNAL protocol. The results obtained were verified to be the same with the margins calculated using an Excel spreadsheet. The algorithm calculates the weekly offline setup error correction values automatically and reduces the risk of input data error observed in the spreadsheet. CONCLUSIONS: In conclusion, the algorithm provides an automated method for optimization and reduction of PTV margin using logged setup errors from CBCT-based IGRT.

Spatiotemporal nexus between vegetation change and extreme climatic indices and their possible causes of change.

Climate extremes have a significant impact on vegetation. However, little is known about vegetation response to climatic extremes in Bangladesh. The association of Normalized Difference Vegetation Index (NDVI) with nine extreme precipitation and temperature indices was evaluated to identify the nexus between vegetation and climatic extremes and their associations in Bangladesh for the period 1986-2017. Moreover, detrended fluctuation analysis (DFA) and Morlet wavelet analysis (MWA) were employed to evaluate the possible future trends and decipher the existing periodic cycles, respectively in the time series of NDVI and climate extremes. Besides, atmospheric variables of ECMWF ERA5 were used to examine the casual circulation mechanism responsible for climatic extremes of Bangladesh. The results revealed that the monthly NDVI is positively associated with extreme rainfall with spatiotemporal heterogeneity. Warm temperature indices showed a significant negative association with NDVI on the seasonal scale, while precipitation and cold temperature extremes showed a positive association with yearly NDVI. The DEA revealed a continuous increase in temperature extreme in the future, while no change in precipitation extremes. NDVI also revealed a significant association with extreme temperature indices with a time lag of one month and with precipitation extreme without time lag. Spatial analysis indicated insensitivity of marshy vegetation type to climate extremes in winter. The study revealed that elevated summer geopotential height, no visible anticyclonic center, reduced high cloud cover, and low solar radiation with higher humidity contributed to climatic extremes in Bangladesh. The nexus between NDVI and climatic extremes established in this study indicated that increasing warm temperature extremes due to global warming might have severe implications on Bangladesh's ecology and the environment in the future.

A homozygous missense variant in the homeobox domain of the NKX6-2 results in progressive spastic ataxia type 8 associated with lower limb weakness and neurological manifestations.

BACKGROUND: Progressive spastic ataxia is a heterogeneous disorder characterized by cerebellar ataxia and limb spasticity associated with other severe neurological complications. Spastic ataxia is classified into pure and complex types, inherited in both an autosomal recessive and autosomal dominant manner. It is caused by pathogenic variants in at least eight different genes, including NKX6-2 (MIM 607063) located on chromosome 10q26.3. The present study aimed to identify the genetic variant(s) underlying progressive spastic ataxia and to establish the genotype-phenotype correlation. METHODS: We collected a large consanguineous family having four affected individuals segregating progressive spastic ataxia in an autosomal recessive manner. To investigate the molecular cause of the disease, genomic DNA of three affected individuals underwent whole exome sequencing. RESULTS: All of the affected individuals showed progressive clinical features such as spastic ataxia, lower limb weakness and other mild neurological abnormalities. Whole exome sequencing data were analyzed using different filters. Filtering of rare and shared homozygous variants revealed a novel homozygous missense variant (c.545C>T; p.Ala182Val) in a highly conserved homeobox domain of the NKX6-2 protein. CONCLUSIONS: The findings of the present study add a novel variant to the NKX6-2 mutation spectrum and provide evidence that homozygous variants in the NKX6-2 cause progressive spastic ataxia associated with other abnormalities.

The changing sociocultural context of wildlife conservation.

We introduced a multilevel model of value shift to describe the changing social context of wildlife conservation. Our model depicts how cultural-level processes driven by modernization (e.g., increased wealth, education, and urbanization) affect changes in individual-level cognition that prompt a shift from domination to mutualism wildlife values. Domination values promote beliefs that wildlife should be used primarily to benefit humans, whereas mutualism values adopt a view that wildlife are part of one's social network and worthy of care and compassion. Such shifts create emergent effects (e.g., new interest groups) and challenges to wildlife management organizations (e.g., increased conflict) and dramatically alter the sociopolitical context of conservation decisions. Although this model is likely applicable to many modernized countries, we tested it with data from a 2017-2018 nationwide survey (mail and email panel) of 43,949 residents in the United States. We conducted hierarchical linear modeling and correlational analysis to examine relationships. Modernization variables had strong state-level effects on domination and mutualism. Higher levels of education, income, and urbanization were associated with higher percentages of mutualists and lower percentages of traditionalists, who have strong domination values. Values affected attitudes toward wildlife management challenges; for example, states with higher proportions of mutualists were less supportive of lethal control of wolves (Canis lupus) and had lower percentages of active hunters, who represent the traditional clientele of state wildlife agencies in the United States. We contend that agencies will need to embrace new strategies to engage and represent a growing segment of the public with mutualism values. Our model merits testing for application in other countries.

El Cambiante Contexto Sociocultural de la Conservación de Fauna Resumen Introdujimos un modelo multinivel del cambio de valores para describir el cambiante contexto social de la conservación de fauna. Nuestro modelo representa cómo los procesos a nivel cultural llevados por la modernización (p. ej.: aumento de riqueza, educación y urbanización) afectan a los cambios en la cognición a nivel individual que incitan a un cambio de los valores de dominación a los valores de mutualismo de la fauna. Los valores de dominación promueven la creencia de que la fauna debería usarse principalmente para beneficio de los humanos, mientras que los valores de mutualismo adoptan una visión de que la fauna es parte de la red social de uno y digna de cuidados y compasión. Dichos cambios generan efectos emergentes (p. ej.: nuevos grupos de interés) y retos para las organizaciones de manejo de fauna (p. ej.: conflictos mayores) y alteran dramáticamente el contexto sociopolítico de las decisiones de conservación. Aunque este modelo probablemente pueda aplicarse a muchos países modernizados, lo pusimos a prueba con datos de un censo nacional de 2017 - 2018 realizado (por correo y correo electrónico) a 43,949 residentes de los Estados Unidos. Realizamos un modelado jerárquico lineal y un análisis de correlación para examinar las relaciones. Las variables de modernización tuvieron efectos sólidos a nivel estatal sobre la dominación y el mutualismo. Los niveles altos de educación, ingresos y urbanización estuvieron asociados con los porcentajes más altos de mutualistas y con los porcentajes más bajos de tradicionalistas, quienes tienen valores de dominación fuertes. Los valores afectaron a las actitudes hacia los retos para el manejo de fauna; por ejemplo, los estados con proporciones mayores de mutualistas mostraron un menor apoyo para el control letal de los lobos (Canis lupus) y tuvieron porcentajes más bajos de cazadores activos, quienes representan a la clientela tradicional de las agencias estatales de vida silvestre en los Estados Unidos. Sostenemos que las agencias necesitarán adoptar nuevas estrategias para envolver y representar a un segmento creciente del público con valores mutualistas. Nuestro modelo amerita ser evaluado para su aplicación en otros países.

A novel missense variant in the RASGRP2 gene in patients with moderate to severe bleeding disorder.

Inherited platelet function disorder-18 (IPD-18) is a relatively new non-syndromic autosomal recessive bleeding disorder. It is characterized by deficient or dysfunctional CalDAG-GEFI protein. The distinctive feature of the disease is impaired platelet aggregation in response to multiple physiologic agonists. We here report a family with a platelet-type bleeding disorder and a novel mutation in the RASGRP2 gene. The overall bleeding score for the affected individuals was 15 and 12. Based on the initial diagnosis of Glanzmann thrombasthenia, targeted sequencing of integrin subunit alpha 2b and integrin subunit beta 3 encoding genes ITGA2B and ITGB3 were carried out in both affected members of a family. Sequence alignment failed to identify the disease-causing variant(s) in both genes. Therefore, whole exome sequencing in one affected individual was performed. Data analysis detected a novel homozygous missense variant (c.956C>T; p.Pro319Leu) in the exon 9 of the RASGRP2 gene. Five additional individuals of a family including both parents, an affected individual and two asymptomatic individuals were Sanger sequenced for the variant (c.956C>T). The variant segregates in the family in an autosomal recessive manner. Several in silico tools predicted the variant as pathogenic. Protein modeling studies suggest that the mutation (p.Pro319Leu) cause a conformational change in the loop structure of the RasGEF domain of the CalDAG-GEFI protein. Reported variants in the RasGEF domain impair expression and/or nucleotide exchange activity of CalDAG-GEFI protein and thus inhibit the activation of Rap1 protein required for platelet adhesion and hemostatic plug formation.

Genetic variations in autoimmune genes and VKH disease.

PURPOSE: Vogt-Koyanagi-Harada (VKH) disease is a rare autoimmune disease. The autoimmune response in VKH disease is against the melanin-producing cells; therefore, in affected individuals melanocyte-containing organs manifest disease symptoms including eyes, ears, skin and nervous system. VKH is a multifactorial disease, and the precise cause of the VKH disease is unknown. Studies have suggested that both environmental and genetic factors are responsible for the VKH disease. In this review, the authors have collected all the available literature on the genetics of VKH to their knowledge and discussed the role of genetic variants in causing VKH disease. METHODS: An extensive literature search was performed in order to review all the published studies regarding VKH clinical phenotyping and genetic variants in VKH disease. Medline, PubMed, Cochrane library, and Scopus was searched using combination of keywords. RESULTS: It was found that variants in HLA genes, IL-12b, TNFSF4, and miR-20-5p genes are significantly associated with VKH; however, variants in genes ATG10, TNIP1 and CLEC16A did not achieve significant genome-wide association threshold. Moreover, polymorphisms in TNIP1 and CLEC16A play a protective role against VKH. CONCLUSION: The authors conclude that increased sample size and a more homogeneous VKH patient population may reveal a significant association of variants in ATG10, TNIP1 and CLEC16A genes with VKH disease.

Exome sequencing identified rare variants in genes HSPG2 and ATP2B4 in a family segregating developmental dysplasia of the hip.

BACKGROUND: Developmental dysplasia of the hip (DDH) is a common pathological condition of the musculoskeletal system in infants which results in a congenital and developmental malformation of the hip joint. DDH is a spectrum of pathologies affecting the infant hip ranging from asymptomatic subtle radiographic signs through mild instability to frank dislocations with acetabular dysplasia. A Saudi family with three affected individuals with DDH was identified and genetic analysis was performed to detect the possible genetic defect(s) underlying DDH in the affected members of the family. METHODS: We performed whole genome genotyping using Illumina HumanOmni 2.5 M array and whole exome sequencing (WES) using Nextera Rapid capture kit and Illumina NextSeq500 instrument in four individuals of a family with DDH. RESULTS: SNP data analysis did not identify any runs of homozygosity and copy number variations. Identity-by-descent (IBD) analysis on whole genome genotyping data identified a shared haplotypes on chromosome 1 in affected individuals. An analysis of the WES data identified rare heterozygous variants in HSPG2 and ATP2B4 genes in the affected individuals. Multiple prediction software predicted that the variants identified are damaging. Moreover, in silico analysis showed that HSPG2 regulates ATP2B4 expression using a variety of transcription factors. CONCLUSION: Our results indicate that there might be a functional epistatic interaction between HSPG2 and ATP2B4, and DDH in the family studied is due to a combined effect of both variants. These variants are also present in the asymptomatic mother suggesting that the variants in HSPG2 and ATP2B4 are incompletely penetrant. This study provides the first evidence of digenic inheritance of DDH in a family and extends the spectrum of genetic heterogeneity in this human disorder.

Immunohistochemical staining of leptin is associated with grade, stage, lymph node involvement, recurrence, and hormone receptor phenotypes in breast cancer.

BACKGROUND: Obesity is part of the established risk factors for breast cancer (BC) in postmenopausal females. Circulating leptin increases in parallel with the increase of body weight and fat reservoir. METHODS: This research investigated the link between leptin phenotype and the clinicopathological factors in BC. A large set of breast cancer cases (449), and 27 non-cancerous tissue samples of breast were employed for leptin expression recognition using immunohistochemistry staining. RESULTS: Cytoplasmic immunohistochemical staining of leptin was recognized in 376 (83.7%) and 25 (92.6%) of BC and control cases respectively. Leptin immunostaining were significantly associated with age, histotypes, grade, stage, lymph node involvement, tumor recurrence, hormone receptor phenotypes, ER and HER2 expressions, and p-values were (P = 0.0233), (P = 0.0001), (P = 0.050), (P = 0.0291), (P = 0.0300), (P = 0.0023), (P = 0.0021), (P = 0.0279) respectively. Reasonable proportion of cases with low staining score was more prevalent in all subgroups of clinicopathological parameters except ER- PR+ HER2- hormone receptor phenotype and mucinous carcinoma which showed high level of leptin immunoreactivity. Tumor recurrence is less prevailing in high score leptin immunostaining cases. Furthermore, Log Rank (Mantel-Cox) test findings revealed considerably different survival distributions were observed for the different categories of leptin immunostaining scores (P = 0.032). Negative leptin immunostaining is related to poor survival. CONCLUSIONS: Our preliminary findings support leptin clinical value in confirming BC diagnosis as well as prognosis. These results suggest that leptin molecule is an important biomarker that could identify type, grade, stage, lymph node involvement, relapse and prognosis in breast cancer.

CIT, a gene involved in neurogenic cytokinesis, is mutated in human primary microcephaly.

Autosomal recessive primary microcephaly (MCPH) is a static neurodevelopmental disorder characterized by congenital small head circumference and non-progressive intellectual disability without additional severe brain malformations. MCPH is a genetically heterogeneous disorder. Sixteen genes (MCPH1-MCPH16) have been discovered so far, mutations thereof lead to autosomal recessive primary microcephaly. In a family, segregating MCPH in an autosomal recessive manner, genome-wide homozygosity mapping mapped a disease locus to 16.9-Mb region on chromosome 12q24.11-q24.32. Following this, exome sequencing in three affected individuals of the family discovered a splice site variant (c.753+3A>T) in citron kinase (CIT) gene, segregating with the disorder in the family. CIT co-localizes to the midbody ring during cytokinesis, and its loss of expression results in defects in neurogenic cytokinesis in both humans and mice. Splice site variant in CIT, identified in this study, is predicted to abolish splice donor site. cDNA sequence of an affected individual showed retention of an intron next to the splice donor site. The study, presented here, revealed the first variant in the CIT causing MCPH in the family.

Xq21.31-q21.32 duplication underlies intellectual disability in a large family with five affected males.

X-linked intellectual disability is the most common form of neurological disorder in male and accounts for 5-10% of incidence in the population. Copy number variants (CNVs) have been studied extensively to identify genomic regions responsible for neurological disorders. Array CGH and SNP genotyping have identified several CNVs on X-chromosome in patients with X-linked intellectual disability. We genotyped 2.5 million SNPs in 10 individuals of a 4 generation family segregating X-linked intellectual disability using Illumina Infinium BeadChip assay. Whole genome genotyping data analysis identified a single duplication of 3.95 Mb on X-chromosome in all five affected male individuals. This CNV is inherited from a healthy mother. All five affected individuals manifest moderate to severe intellectual disability, seizures and behavioral abnormalities. X-chromosome inactivation analysis showed that X-chromosome of the mother with duplication is completely inactivated which has also been found in daughters.