Miracle-dz/UniFAR-FTUs huggingface dataset:
The role of MHC class I recycling and Arf6 in cross-presentation by murine dendritic cells

MHC-I internalization and recycling in DCs neither requires Arf6 nor provides MHC-I to the Rab11+ endocytic recycling compartment; however, cross-presentation of immune complexes depends on Arf6. Cross-presentation by MHC class I molecules (MHC-I) is critical for priming of cytotoxic T cells. Peptides derived from cross-presented antigens can be loaded on MHC-I in the endoplasmic reticulum and in endocytic or phagocytic compartments of murine DCs. However, the origin of MHC-I in the latter compartments is poorly understood. Recently, Rab22-dependent MHC-I recycling through a Rab11+ compartment has been suggested to be implicated in cross-presentation. We have examined the existence of MHC-I recycling and the role of Arf6, described to regulate recycling in nonprofessional antigen presenting cells, in murine DCs. We confirm folded MHC-I accumulation in a juxtanuclear Rab11+ compartment and partially localize Arf6 to this compartment. MHC-I undergo fast recycling, however, both folded and unfolded internalized MHC-I fail to recycle to the Rab11+Arf6+ compartment. Therefore, the source of MHC-I molecules in DC endocytic compartments remains to be identified. Functionally, depletion of Arf6 compromises cross-presentation of immune complexes but not of soluble, phagocytosed or mannose receptor–targeted antigen, suggesting a role of Fc receptor–regulated Arf6 trafficking in cross-presentation of immune complexes.
Automated Analysis of Platelet Aggregation on Cultured Endothelium in a Microfluidic Chip Perfused with Human Whole Blood

Organ-on-a-chip models with incorporated vasculature are becoming more popular to study platelet biology. A large variety of image analysis techniques are currently used to determine platelet coverage, ranging from manually setting thresholds to scoring platelet aggregates. In this communication, an automated methodology is introduced, which corrects misalignment of a microfluidic channel, automatically defines regions of interest and utilizes a triangle threshold to determine platelet coverages and platelet aggregate size distributions. A comparison between the automated methodology and manual identification of platelet aggregates shows a high accuracy of the triangle methodology. Furthermore, the image analysis methodology can determine platelet coverages and platelet size distributions in microfluidic channels lined with either untreated or activated endothelium used for whole blood perfusion, proving the robustness of the method.
Standing at work and preterm delivery

Objective Guidelines in some European countries and the United States suggest that pregnant women should avoid prolonged standing and heavy lifting in the workplace during the second and third trimester of pregnancy. However, results from epidemiological studies on this topic are ambiguous. The aim of this study was to evaluate the influence of standing and walking at work in the second trimester on preterm delivery in a population with a low frequency of other workplace hazards.
Prognostic profiling of the immune cell microenvironment in Ewing´s Sarcoma Family of Tumors

ABSTRACT Ewing´s Sarcoma Family of Tumors (ESFT) are clinically aggressive bone and soft tissue tumors in children and young adults. Analysis of the immune tumor microenvironment (TME) provides insight into tumor evolution and novel treatment options. So far, the scarcity of immune cells in ESFT has hindered a comprehensive analysis of rare subtypes. We determined the relative fraction of 22 immune cell types using 197 microarray gene expression datasets of primary ESFT tumor samples by using CIBERSORT, a deconvolution algorithm enumerating infiltrating leucocytes in bulk tumor tissue. The most abundant cells were macrophages (mean 43% of total tumor-infiltrating leukocytes, TILs), predominantly immunosuppressive M2 type macrophages, followed by T cells (mean 23% of TILs). Increased neutrophils, albeit at low number, were associated with a poor overall survival (OS) (p = .038) and increased M2 macrophages predicted a shorter event-free survival (EFS) (p = .033). High frequency of T cells and activated NK cells correlated with prolonged OS (p = .044 and p = .007, respectively). A small patient population (9/32) with combined low infiltrating M2 macrophages, low neutrophils, and high total T cells was identified with favorable outcome. This finding was confirmed in a validation cohort of patients with follow up (11/38). When comparing the immune TME with expression of known stemness genes, hypoxia-inducible factor 1 α (HIF1α) correlated with high abundance of macrophages and neutrophils and decreased T cell levels. The immune TME in ESFTs shows a distinct composition including rare immune cell subsets that in part may be due to expression of HIF1α.
Modulation of hepatocyte sialylation drives spontaneous fatty liver disease and inflammation.

Circulatory protein glycosylation is a biomarker of multiple disease and inflammatory states and has been applied in the clinic for liver dysfunction, heart disease, and diabetes. With the notable exception of antibodies, the liver produces most of the circulatory glycoproteins, including the acute phase proteins released as a function of the inflammatory response. Among these proteins is β-galactoside α2,6-sialyltransferase (ST6Gal1), an enzyme required for α2,6-linked sialylation of glycoproteins. Here, we describe a hepatocyte-specific conditional knockout of ST6Gal1 (H-cKO) using albumin promoter-driven Cre-lox recombination. We confirm the loss of circulatory glycoprotein α2,6 sialylation and note no obvious dysfunction or pathology in young H-cKO mice, yet these mice show robust changes in plasma glycoprotein fucosylation, branching, and the abundance of bisecting GlcNAc and marked changes in a number of metabolic pathways. As H-cKO mice aged, they spontaneously developed fatty liver disease characterized by the buildup of fat droplets in the liver, inflammatory cytokine production, and a shift in liver leukocyte phenotype away from anti-inflammatory Kupffer cells and towards pro-inflammatory M1 macrophages. These findings connect hepatocyte and circulatory glycoprotein sialylation to the regulation of metabolism and inflammation, potentially identifying the glycome as a new target for liver-driven disease.
Genotyping of viable Toxoplasma gondii from the first national survey of feral swine revealed evidence for sylvatic transmission cycle, and presence of highly virulent parasite genotypes

Abstract Feral swine are known reservoirs of various pathogens, including Toxoplasma gondii. Here, we report the first national survey of viable T. gondii in feral swine in the USA. We paired serological surveys with parasite isolation and bioassay to evaluate the prevalence and genetic diversity of these parasites. From 2012–2017, sera and tissues from 1517 feral swine across the USA were collected for the isolation of viable T. gondii. Serum samples were initially screened for antibodies to T. gondii, and then the tissues of seropositive feral swine were bioassayed in mice. Antibodies were detected in 27.7% of feral swine tested by the modified agglutination test (1:25 or higher). Antibody positive rates increased significantly with age, with 10.1% of juveniles, 16.0% of sub-adults and 38.4% of adults testing seropositive. Myocardium (50 g) from 232 seropositive feral swine was digested in pepsin and bioassayed in mice. Viable T. gondii was isolated from 78 feral swine from 21 states. Twelve of the 78 isolates were pathogenic to outbred Swiss Webster mice and 76 of the 78 isolates could be propagated further in cell culture and were genotyped. For genotyping, deoxyribonucleic acid extracted from cell culture-derived tachyzoites was characterized by polymerase chain reaction restriction fragment length polymorphism using the genetic markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genotyping revealed 15 ToxoDB genotypes, including 43 isolates for genotype #5 (haplogroup 12), 11 isolates for #24, four isolates for #2 (haplogroup 3), two isolates for each of genotypes #3 (haplogroup 2), #4 (haplogroup 12), #216, #221, #289 and #297 and one isolate for each of genotypes #1 (haplogroup 2), #39, #66, #260, #261 and #299. Genotype #5 was the most frequently isolated, accounted for 57% (43/76) of the isolates, followed by #24, accounted for 14% (11/76). Genotypes #260, #289, #297 and #299 are new types. Genotype #289 was highly virulent to mice and originated from feral swine collected in Louisiana on the same day at the same location. Genotype #216 was previously demonstrated to be highly virulent to mice. Our results indicate moderate genetic diversity of T. gondii in feral swine in the USA, with the genotype #5 (haplogroup 12) dominant in the continental USA, whereas genotype #24 (10/14) was dominant in Hawaii, suggesting different population structures of the parasites among the two distinct geographical locations.
Male Homosexual Preference: Femininity and the Older Brother Effect in Indonesia

Male homosexual preference (MHP) is an evolutionary enigma because it is partially heritable and imposes a fertility cost. In occidental societies, homosexual men are feminized at various levels and they have more older brothers than heterosexual men. To evaluate whether femininity and the fraternal birth order (FBO) effect are universal features of MHP or not, we collected original data from homosexual men, heterosexual men, and heterosexual women from Java (Indonesia). Facial photographs were used to test whether homosexual faces are feminized when compared with heterosexual ones. We found that faces manipulated to resemble the average face of homosexual men are perceived as facially feminized, suggesting that homosexual men are facially feminized compared to heterosexual men, although a higher facial femininity was not captured by morphological analyses. Then, family data were used to detect differences in siblings’ composition between homosexuals and heterosexuals. Homosexual men displayed a higher number of older brothers than heterosexual men, even when sibship size was controlled for, suggesting that the FBO effect exists in Indonesian populations. Independent of sexual orientation, men with older brothers seem more feminized than those without older brothers, consistent with the immune origin of the FBO effect. In conclusion, MHP in Indonesia is partially feminized and they have more older brothers. Such features are also associated with MHP in other cultural contexts, suggesting a cross-cultural effect of men homosexual preference. An evolutionary explanation is available for the feminizing effect, although the FBO effect remains unexplained even if proximal mechanisms start to be identified.
Proof of concept for identifying cystic fibrosis from perspiration samples

Significance Early diagnosis and characterization of the severity of CFTR mutations carried in cystic fibrosis (CF) impacts life expectancy and quality of life for patients. We demonstrate a testing platform that combines analysis of perspiration samples by desorption electrospray ionization mass spectrometry and a machine-learning method of gradient boosted decision trees, with an accuracy for the correct identification of CF cases of 98 ± 2%. Our sampling method is minimally invasive; it only requires swiping a standard microscope slide across the patient’s forehead, with no sample processing. The whole collection and testing process takes less than 2 min, which suggests a faster alternative with comparable accuracy to the conventional sweat chloride test, which takes up to 3 h. The gold standard for cystic fibrosis (CF) diagnosis is the determination of chloride concentration in sweat. Current testing methodology takes up to 3 h to complete and has recognized shortcomings on its diagnostic accuracy. We present an alternative method for the identification of CF by combining desorption electrospray ionization mass spectrometry and a machine-learning algorithm based on gradient boosted decision trees to analyze perspiration samples. This process takes as little as 2 min, and we determined its accuracy to be 98 ± 2% by cross-validation on analyzing 277 perspiration samples. With the introduction of statistical bootstrap, our method can provide a confidence estimate of our prediction, which helps diagnosis decision-making. We also identified important peaks by the feature selection algorithm and assigned the chemical structure of the metabolites by high-resolution and/or tandem mass spectrometry. We inspected the correlation between mild and severe CFTR gene mutation types and lipid profiles, suggesting a possible way to realize personalized medicine with this noninvasive, fast, and accurate method.
ONE HUNDRED MILLION HEALTHIER LIVES: MEASURING WHAT MATTERS

Abstract The rise of chronic disease, an aging population, inequity in health care access and outcomes, and new technology demand a fundamentally different approach to fostering health—one that brings sectors together to address the social and behavioral determinants of health. To address these concerns, NCOA is leading the Aging Hub of the 100 Million Healthier Lives (100MLives), an initiative convened by the Institute for Healthcare Improvement. 100MLives is a collaboration of change agents across sectors working to transform how the world thinks and acts to improve health, well-being, and equity across the lifespan. NCOA is collaborating with national and local organizations, as well as federal agencies to support a nationwide ecosystem for measuring, fostering, and bringing to scale innovations that address social and behavioral determinants of health. The Aging Hub is committed to consistently measuring what matters (adult well-being assessment-AWA), learn what programs and services within the aging network improve a person’s life, and work with partners to scale those programs. The initiative encourages use of the Adult Well-Being Assessment, a validated 7-question survey that asks individuals to rate their overall well-being; financial, physical, mental, and spiritual health; and social support. Shared use of these outcome measures can make it easier for the aging network to demonstrate the impact of their programs and services. Join NCOA to learn about progress to date, including a report on AWA and demographic data collected by the Baltimore County Department with over 10,000 older adults and how data is being used to improve service delivery.
Panorama epidemiológico de la mortalidad por cáncer en el Instituto Mexicano del Seguro Social: 1991-1995

Objective. This paper describes the global cancer mortality and the specific mortality patterns for the main neoplasms among adult members of the Mexican Institute of Social Security (IMSS). Material and methods. Using official death certificates and information about the population of the IMSS members during 1991-1995, national and regional annual global cancer mortality as well as specific mortality rates for the 10 most important malignant neoplasms by sex were estimated among people older than 20 years of age. The trends for these neoplasms during the study period were estimated by means of Poisson regression. The rate differences in specific cancer mortality by region and sex, for the two major neoplasms, were calculated subtracting specific regional rates from the respective national rate in 1995. Results. The global mortality rate for cancer among men increased from 76.2 in 1991 to 94.8 x 100 000 IMSS´members in 1995; and among women from 85.6 to 105.8 x 100 000 IMSS´members, representing an increment of 24.4 and 24% men and women, respectively, during the study period. Among men, neoplasm of kidney, leukemia, pancreas, prostate and lung showed the major increment; among women, neoplasm of colon, breast, pancreas, leukemia and liver showed the most significant increment. Conclusions. In the IMSS it is necessary the integration of a population based cancer registry. The registry will play a main role in disease surveillance and control; will give basic information over incidence and temporal variation, and could be the main source of information for epidemiologic research, as well as planning and evaluation of the quality of medical attention services such as prevention and early diagnosis and treatment.
C. elegans Runx/CBFβ suppresses POP-1 TCF to convert asymmetric to proliferative division of stem cell-like seam cells

ABSTRACT A correct balance between proliferative and asymmetric cell divisions underlies normal development, stem cell maintenance and tissue homeostasis. What determines whether cells undergo symmetric or asymmetric cell division is poorly understood. To gain insight into the mechanisms involved, we studied the stem cell-like seam cells in the Caenorhabditis elegans epidermis. Seam cells go through a reproducible pattern of asymmetric divisions, instructed by divergent canonical Wnt/β-catenin signaling, and symmetric divisions that increase the seam cell number. Using time-lapse fluorescence microscopy we observed that symmetric cell divisions maintain asymmetric localization of Wnt/β-catenin pathway components. Our observations, based on lineage-specific knockout and GFP-tagging of endogenous pop-1, support the model that POP-1TCF induces differentiation at a high nuclear level, whereas low nuclear POP-1 promotes seam cell self-renewal. Before symmetric division, the transcriptional regulator RNT-1Runx and cofactor BRO-1CBFβ temporarily bypass Wnt/β-catenin asymmetry by downregulating pop-1 expression. Thereby, RNT-1/BRO-1 appears to render POP-1 below the level required for its repressor function, which converts differentiation into self-renewal. Thus, we found that conserved Runx/CBFβ-type stem cell regulators switch asymmetric to proliferative cell division by opposing TCF-related transcriptional repression. Summary: To switch asymmetric to proliferative cell division, the C. elegans RNT-1/BRO-1 transcriptional repressor opposes POP-1 TCF expression in seam stem cells, which turns POP-1-induced differentiation into self-renewal.
West Nile Virus in Wildlife and Nonequine Domestic Animals, South Africa, 2010–2018

West Nile virus (WNV) lineage 2 is associated with neurologic disease in horses and humans in South Africa. Surveillance in wildlife and nonequine domestic species during 2010–2018 identified WNV in 11 (1.8%) of 608 animals with severe neurologic and fatal infections, highlighting susceptible hosts and risk for WNV epizootics in Africa.
ACP-TX-I and ACP-TX-II, Two Novel Phospholipases A2 Isolated from Trans-Pecos Copperhead Agkistrodon contortrix pictigaster Venom: Biochemical and Functional Characterization

This work reports the purification and biochemical and functional characterization of ACP-TX-I and ACP-TX-II, two phospholipases A2 (PLA2) from Agkistrodon contortrix pictigaster venom. Both PLA2s were highly purified by a single chromatographic step on a C18 reverse phase HPLC column. Various peptide sequences from these two toxins showed similarity to those of other PLA2 toxins from viperid snake venoms. ACP-TX-I belongs to the catalytically inactive K49 PLA2 class, while ACP-TX-II is a D49 PLA2, and is enzymatically active. ACP-TX-I PLA2 is monomeric, which results in markedly diminished myotoxic and inflammatory activities when compared with dimeric K49 PLA2s, confirming the hypothesis that dimeric structure contributes heavily to the profound myotoxicity of the most active viperid K49 PLA2s. ACP-TX-II exhibits the main pharmacological actions reported for this protein family, including in vivo local myotoxicity, edema-forming activity, and in vitro cytotoxicity. ACP-TX-I PLA2 is cytotoxic to A549 lung carcinoma cells, indicating that cytotoxicity to these tumor cells does not require enzymatic activity.
Impact of posterior septum resection on nasal airflow pattern and warming function

Abstract Background: The functional significance of the posterior segment of the septum is not fully understood. Aims/Objectives: The purpose of this study was to evaluate the impact of posterior septum resection with varying extents on nasal airflow pattern and warming function, and to determine the appropriate resection extent on posterior septum without disturbing airflow pattern. Material and Methods: Three dimensional models were established from the nasal CT scan of a healthy female volunteer without nasal disease. The models included the normal, as well as 1 cm, 2 cm, and 3 cm extent of posterior septum resection by virtual surgery. Using computational fluid dynamics, the airflow characteristics such as average and peak airflow velocity, average wall shear stress, average temperature and turbulence kinetic energy were recorded and compared among the four models. Results: By comparison with the normal model, the virtual surgeries with varying extents of posterior septum resection (1 cm, 2 cm, and 3 cm), demonstrated little alteration on indices of the average and peak airflow velocity, average wall shear stress, airflow temperature and turbulence kinetic energy. However, a statistical difference of the abovementioned indices existed when compared between varying planes of the nasal cavity (p < .001), although this did not appear to be significantly influenced by the degree of posterior septectomy. Conclusions and significance: From an aerodynamics perspective, little disturbance on airflow pattern and warming function of nasal cavity could be detected with increasing resection of posterior septum (up to 3 cm). These results may imply that resection of the posterior septum (up to 3 cm) has little impact on nasal function.
The construction of learning curves for basic skills in anesthetic procedures: an application for the cumulative sum method.

This study aimed at constructing learning curves for basic procedural skills in anesthesiology using the cumulative sum method. We recorded 1234 peripheral venous cannulations, 895 orotracheal intubations, 688 spinals, and 344 epidurals performed by residents during the first 10 mo of training. Lear
CRISPR/Cas9-Mediated Knock-Out of KrasG12D Mutated Pancreatic Cancer Cell Lines

In 90% of pancreatic ductal adenocarcinoma cases, genetic alteration of the proto-oncogene Kras has occurred, leading to uncontrolled proliferation of cancerous cells. Targeting Kras has proven to be difficult and the battle against pancreatic cancer is ongoing. A promising approach to combat cancer was the discovery of the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system, which can be used to genetically modify cells. To assess the potential of a CRISPR/CRISPR-associated protein 9 (Cas9) method to eliminate Kras mutations in cells, we aimed to knock-out the c.35G>A (p.G12D) Kras mutation. Therefore, three cell lines with a heterozygous Kras mutation (the human cell lines SUIT-2 and Panc-1 and the cell line TB32047 from a KPC mouse model) were used. After transfection, puromycin selection and single-cell cloning, proteins from two negative controls and five to seven clones were isolated to verify the knock-out and to analyze changes in key signal transduction proteins. Western blots showed a specific knock-out in the KrasG12D protein, but wildtype Kras was expressed by all of the cells. Signal transduction analysis (for Erk, Akt, Stat3, AMPKα, and c-myc) revealed expression levels similar to the wildtype. The results described herein indicate that knocking-out the KrasG12D mutation by CRISPR/Cas9 is possible. Additionally, under regular growth conditions, the knock-out clones resembled wildtype cells.