Miracle-dz/UniFAR-FTUs huggingface dataset:
Failure to Generate Comparable International Normalized Ratio Values Using Five Different Thromboplastin Reagents in Parallel Studies of Patients Receiving Warfarin

Warfarin therapy has traditionally been monitored using the prothrombin time (PT). A significant problem with this assay is the variable sensitivity of commercially available thromboplastin reagents to reduction of vitamin K-dependent coagulation factors. The International Normalized Ratio (INR) was developed as a means of standardizing PT values between laboratories that use different thromboplastins and types of instrumentation. Parallel testing of samples from 63 patients stabilized on oral anticoagulation with five different thromboplastins was undertaken. Forty-eight percent of all samples had INR values that were not identical but showed good correlation. Fifty-two percent of the samples had clinically significant discrepancies of their INR values. In this group, patients in the therapeutic range with one thromboplastin appeared over- or underanticoagulated based on the INR using a different thromboplastin. In order to determine whether use of a low-ISI thromboplastin reagent could provide more reproducible INR results, concurrent testing of specimens from 36 patients on stable oral anticoagulation was undertaken between our hospital laboratory and another, nonaffiliated institution. Both hospitals used similar instrumentation and thromboplastin reagents. Under these conditions, the INR values generated between laboratories correlated highly. The inability to generate consistent INR values in parallel testing using different thromboplastins with identical samples and instrumentation raises concern about the reliability of the INR, and suggests that further analysis is necessary to identify the source of these discrepancies. In the interim, our data show use of a recombinant thromboplastin with a low-ISI value substantially improves interlaboratory variation in INR value. Key Words: International Normalized Ratio—Prothrombin time—Thromboplastin—Anticoagulation monitoring.
A Single Amino Acid Change in Inhibitory Killer Cell Ig-like Receptor Results in Constitutive Receptor Self-Association and Phosphorylation

Signaling by immunoreceptors is often initiated by phosphorylation of cytosolic tyrosines, which then recruit effector molecules. In the case of MHC class I–specific inhibitory receptors, phosphorylation of cytosolic tyrosine residues within ITIMs results in recruitment of a protein tyrosine phosphatase that blocks activation signals. Recent work showed that signaling by an HLA-C–specific killer cell Ig-like receptor (KIR) is independent of signaling by activation receptors. It is not known how ITIM phosphorylation is initiated and regulated. In this article, we show that substitution of His-36 in the first Ig domain of KIR2DL1 with alanine (KIR2DL1-H36A) resulted in constitutive KIR2DL1 self-association and phosphorylation, as well as recruitment of tyrosine phosphatase SHP-1. Furthermore, substitution of His-36 with a similar bulky amino acid, phenylalanine, maintained the receptor in its unphosphorylated state, suggesting that steric hindrance by the His-36 side chain prevents constitutive KIR2DL1 self-association and ITIM phosphorylation. The equally strong phosphorylation of KIR2DL1 and KIR2DL1-H36A after inhibition of tyrosine phosphatase by pervanadate suggested that KIR2DL1-H36A is selectively protected from dephosphorylation. We propose that KIR phosphorylation is controlled by the accessibility of ITIM to tyrosine phosphatases and that KIR binding to HLA-C must override the hindrance that His-36 puts on KIR2DL1 self-association. Expression of KIR2DL1-H36A on NK cells led to stronger inhibition of lysis of HLA-C+ target cells than did expression of wild-type KIR2DL1. These results revealed that ITIM phosphorylation is controlled by self-association of KIR and that His-36 serves as a gatekeeper to prevent unregulated signaling through KIR2DL1.
Ex Vivo and In Vitro Effect of Serum Amyloid A in the Induction of Macrophage M2 Markers and Efferocytosis of Apoptotic Neutrophils

Macrophages affect the magnitude and duration of inflammatory response in a functionally heterogeneous manner. The phenotype of macrophages is maintained through a reversible homeostatic mechanism. A number of determinants that modulate macrophage plasticity have been identified, although the precise mechanisms are not fully understood. In this study, we report that stimulation of isolated human blood monocytes and mouse bone marrow–derived macrophages with human serum amyloid A (SAA), a major acute-phase protein, leads to induced expression of macrophage M2 markers, including IL-10, Ym1, Fizz-1, MRC1, IL-1Rn, and CCL17. The same effect was observed with macrophages exposed to SAA in peritoneal cavity. SAA also increases arginase 1 activity and enhances macrophage efferocytosis of apoptotic neutrophils in mouse macrophages. The induction of M2 markers requires MyD88 and the activation of multiple signaling pathways, but it is independent of Stat6. SAA induces IFN regulatory factor (IRF)4 expression and increases its DNA-binding activity. Silencing IRF4 by small interfering RNA abrogates SAA-induced expression of the M2 markers. These results suggest a potential role for SAA to alter macrophage phenotype and modulate macrophage functions through an MyD88-dependent mechanism that involves IRF4-mediated transcription.
Cryptococcus neoformans–Induced Macrophage Lysosome Damage Crucially Contributes to Fungal Virulence

Upon ingestion by macrophages, Cryptococcus neoformans can survive and replicate intracellularly unless the macrophages become classically activated. The mechanism enabling intracellular replication is not fully understood; neither are the mechanisms that allow classical activation to counteract replication. C. neoformans–induced lysosome damage was observed in infected murine bone marrow–derived macrophages, increased with time, and required yeast viability. To demonstrate lysosome damage in the infected host, we developed a novel flow cytometric method for measuring lysosome damage. Increased lysosome damage was found in C. neoformans–containing lung cells compared with C. neoformans–free cells. Among C. neoformans–containing myeloid cells, recently recruited cells displayed lower damage than resident cells, consistent with the protective role of recruited macrophages. The magnitude of lysosome damage correlated with increased C. neoformans replication. Experimental induction of lysosome damage increased C. neoformans replication. Activation of macrophages with IFN-γ abolished macrophage lysosome damage and enabled increased killing of C. neoformans. We conclude that induction of lysosome damage is an important C. neoformans survival strategy and that classical activation of host macrophages counters replication by preventing damage. Thus, therapeutic strategies that decrease lysosomal damage, or increase resistance to such damage, could be valuable in treating cryptococcal infections.
Medical students themselves as surrogate patients increased satisfaction, confidence, and performance in practicing injection skill

Background: Injection skill is one of procedural skills usually taught to medical students. Lack of experience and confidence in their ability creates nervousness, anxiety, and leads to poor performance in giving an injection to patient. Aims: To evaluate the satisfaction on teaching method, perceptions of confidence and feeling of empathy to the patients, and performance in injection skill of medical students toward practicing injection skill using manikin only compared to additional training using themselves as surrogate patients. Methods: Control group consisted of year 5 medical students, who completed studying injection skill from computer assistant instruction, demonstration, and practicing with manikin. The intervention group in addition to those conventional learning methods, directly experienced injection skill using themselves as surrogate patients and received direct feedback from their peers. Both group had a chance to perform injection to actual children and were assessed while performing the injection for their performance of injection procedures. Upon completion of the entire learning process, the students were asked to answer questionnaire presented with Likert-type scales. Results: There were 57 students in the control group and 32 in the intervention group who completed the study. The intervention group reported significantly higher satisfaction on the teaching method, higher level of confidence and empathy to the children who were given injection, and had significantly better performance in preparing the children and giving injection. Both groups are not different in checking accuracy of order, preparing vaccine, selecting injection site, sterile techniques, handling of instruments and injection site, documentation, and explaining to children/parents. Conclusions: Direct experience by medical students themselves as surrogate patients is an appropriate option for learning injection skill and can enhance the student performance, and therefore should be encouraged.
The Minimum Age of Criminal Responsibility: A Medico-Legal Perspective

The minimum age of criminal responsibility is set in England and Wales at 10 years of age (Children and Young Persons Act 1963), whilst the effective age of most civil responsibilities is 16. Research allows us to consider the speed of development of key brain structures in terms of decision-making and this in turn aids in discussion of where the minimum age should be set. This article reviews the evolution of the minimum age of criminal responsibility and examines the setting of this age in light of both civil attainment ages and the current scientific understanding of brain development.
Ability To Develop Broadly Neutralizing HIV-1 Antibodies Is Not Restricted by the Germline Ig Gene Repertoire

The human Ig repertoire is vast, producing billions of unique Abs from a limited number of germline Ig genes. The IgH V region (IGHV) is central to Ag binding and consists of 48 functional genes. In this study, we analyzed whether HIV-1–infected individuals who develop broadly neutralizing Abs show a distinctive germline IGHV profile. Using both 454 and Illumina technologies, we sequenced the IGHV repertoire of 28 HIV-infected South African women from the Centre for the AIDS Programme of Research in South Africa (CAPRISA) 002 and 004 cohorts, 13 of whom developed broadly neutralizing Abs. Of the 259 IGHV alleles identified in this study, approximately half were not found in the International Immunogenetics Database (IMGT). This included 85 entirely novel alleles and 38 alleles that matched rearranged sequences in non-IMGT databases. Analysis of the rearranged H chain V region genes of mAbs isolated from seven of these women, as well as previously isolated broadly neutralizing Abs from other donors, provided evidence that at least eight novel or non-IMGT alleles contributed to functional Abs. Importantly, we found that, despite a wide range in the number of IGHV alleles in each individual, including alleles used by known broadly neutralizing Abs, there were no significant differences in germline IGHV repertoires between individuals who do and do not develop broadly neutralizing Abs. This study reports novel IGHV repertoires and highlights the importance of a fully comprehensive Ig database for germline gene usage prediction. Furthermore, these data suggest a lack of genetic bias in broadly neutralizing Ab development in HIV-1 infection, with positive implications for HIV vaccine design.
Microchip-Based Single-Cell Imaging Reveals That CD56dimCD57−KIR−NKG2A+ NK Cells Have More Dynamic Migration Associated with Increased Target Cell Conjugation and Probability of Killing Compared to CD56dimCD57−KIR−NKG2A− NK Cells

NK cells are functionally educated by self-MHC specific receptors, including the inhibitory killer cell Ig-like receptors (KIRs) and the lectin-like CD94/NKG2A heterodimer. Little is known about how NK cell education influences qualitative aspects of cytotoxicity such as migration behavior and efficacy of activation and killing at the single-cell level. In this study, we have compared the behavior of FACS-sorted CD56dimCD57−KIR−NKG2A+ (NKG2A+) and CD56dimCD57−KIR−NKG2A− (lacking inhibitory receptors; IR−) human NK cells by quantifying migration, cytotoxicity, and contact dynamics using microchip-based live cell imaging. NKG2A+ NK cells displayed a more dynamic migration behavior and made more contacts with target cells than IR− NK cells. NKG2A+ NK cells also more frequently killed the target cells once a conjugate had been formed. NK cells with serial killing capacity were primarily found among NKG2A+ NK cells. Conjugates involving IR− NK cells were generally more short-lived and IR− NK cells did not become activated to the same extent as NKG2A+ NK cells when in contact with target cells, as evident by their reduced spreading response. In contrast, NKG2A+ and IR− NK cells showed similar dynamics in terms of duration of conjugation periods and NK cell spreading response in conjugates that led to killing. Taken together, these observations suggest that the high killing capacity of NKG2A+ NK cells is linked to processes regulating events in the recognition phase of NK–target cell contact rather than events after cytotoxicity has been triggered.
Compensatory Regulatory Networks between CD8 T, B, and Myeloid Cells in Organ Transplantation Tolerance

In transplantation tolerance, numerous regulatory populations have the capacity to inhibit allograft rejection; however, their compensatory capacities have never been clearly evidenced. We have previously demonstrated that the tolerogenic effect mediated by CD8+CD45RClow regulatory T cells (Tregs) in a model of organ transplantation with CD40Ig could be abrogated by permanent depletion of CD8+ cells that resulted in allograft rejection in half of the recipients. This result demonstrated that CD8+ Tregs were essential, but also that half of the recipients still survived indefinitely. We also demonstrated that no other regulatory populations, besides CD8+ Tregs, could induce and maintain allograft tolerance in CD40Ig-treated tolerant animals. In the current study, we analyzed the mechanisms that arose following CD8+ Treg depletion and allowed establishment of networks of new regulatory cells to maintain allograft survival. We identified regulatory B cells (Bregs) and regulatory myeloid cells (RegMCs) as being responsible of the maintenance of the long-term allograft survival. We demonstrated that both regulatory cell subsets efficiently inhibited antidonor immune responses in adoptively transferred recipients. Although Bregs were induced, they were not essential for the maintenance of the graft as demonstrated in IgM-deficient recipients. In addition, we showed that RegMCs were the most suppressive and acted alone, whereas Bregs activity was associated with increased suppressive activity of other subsets in adoptively transferred recipients. Altogether, to our knowledge, we demonstrated in this study for the first time the emergence of both Bregs and RegMCs following Tregs depletion and highlighted the importance of regulatory cell networks and their synergistic potential in transplantation.
Inmunogenicidad y tolerancia de una vacuna contra influenza, en una población mexicana mayor de 55 años de edad

Objective. To confirm the immunogenicity and tolerance of the inactivated, fractionated, and purified influenza vaccine‚ in a Mexican adult population aged 55 and older, medically served at a Petroleos Mexicanos Hospital (Pemex, Mexican Oil Company). Material and Methods. study was conducted between November and December, 2000, among ninety adult subjects aged 55 years and older who were seen at the Hospital Central Sur Pemex. The primary endpoints regarding immunogenicity were the percentage of individuals with protective antibodies targeting hemagglutinins higher than or equal to 1:40, and the percentage of subjects who seroconverted as measured by a four-fold increase in protective antibody production. Secondary endpoints included the frequency of local and systemic reactions to the vaccine. An additional criterion that was evaluated included antigen-antibody affinity assays to measure the polyclonal antibody response to the vaccine and the specific generation of high-affinity antibodies to viral proteins, before and after vaccination. Results. The antibody protection rate was 95.6% against the H1N1 strain, 98.9% against the H3N2 strain, and a 100% against the B/Yamanashi strain. Seroconversion to the H1N1 strain was elicited in 74.4% of subjects, to the H3N2 strain in 88.9%, and to the B / Yama
Species-Specific Minimal Sequence Motif for Oligodeoxyribonucleotides Activating Mouse TLR9

Synthetic oligodeoxyribonucleotides (ODNs) containing unmethylated CpG recapitulate the activation of TLR9 by microbial DNA. ODNs are potent stimulators of the immune response in cells expressing TLR9. Despite extensive use of mice as experimental animals in basic and applied immunological research, the key sequence determinants that govern the activation of mouse TLR9 by ODNs have not been well defined. We performed a systematic investigation of the sequence motif of B class phosphodiester ODNs to identify the sequence properties that govern mouse TLR9 activation. In contrast to ODNs activating human TLR9, where the minimal sequence motif for the receptor activation comprises a pair of closely positioned CpGs we found that the mouse TLR9 requires a single CpG positioned 4–6 nt from the 5′-end. Activation is augmented by a 5′TCC sequence one to three nucleotides from the CG. The distance of the CG dinucleotide of four to six nucleotides from the 5′-end and the ODN’s length fine-tunes activation of mouse macrophages. Length of the ODN <23 and >29 nt decreases activation of dendritic cells. The ODNs with minimal sequence induce Th1-type cytokine synthesis in dendritic cells and confirm the expression of cell surface markers in B cells. Identification of the minimal sequence provides an insight into the sequence selectivity of mouse TLR9 and points to the differences in the receptor selectivity between species probably as a result of differences in the receptor binding sites.
HIV-positive parents’ accounts on disclosure preparation activities for a parent’s and/or a child’s illness in Kenya

The aim of this research brief is to describe a study that examined how HIV-positive parents prepared themselves and their children for HIV disclosure in Kenya. This is the first study from Sub-Saharan Africa (SSA) that provides comprehensive data on how HIVpositive parents prepare themselves and their HIV-positive and negative children for disclosure of a parent’s and/or a child’s illness. Prior studies in SSA have provided limited details about the activities performed by parents to prepare for disclosure of a parent’s or a child’s illness. Key aspects of preparing for disclosure to children: 1. Most parents take years to prepare for disclosure, proceeding when they judge themselves ready to impart the news and their children receptive to receive the news. 2. Parents’ preparation activities for disclosure proceed through four major phases which include secrecy, exploration, readiness, and finally full disclosure of illness. 3. In the secrecy phase parents do not disclose; in the exploration phase they plan how they will disclose; in the readiness phase they seek activities that will help them to fully disclose; finally when ready they fully disclose to their children based on birth order. 4. Parents who have many children remain simultaneously within the different preparation phases as they move their children from a state where none are disclosed to, to a state when all of them have been fully disclosed to. The original research article is located at: http://scholarworks.waldenu. edu/jsbhs/vol8/iss1/1/
Regulation of bone resorption and mineral homeostasis by osteocytes

Osteocytes are the most abundant cells in bone, but their relative inaccessibility and (until recently) a lack of good in vitro and in vivo models have impeded progress in understanding their functional roles. Compelling evidence points to a mechanosensory function whereby osteocytes regulate bone modeling and remodeling in response to shear or strain forces, but their possible role in calcium and phosphate homeostasis is less clear. Moreover, recent work indicates that the ablation of osteocytes leads to the development of osteoporosis, suggesting that these cells are involved in normal bone development and/or remodeling. Several paracrine and endocrine factors, notably parathyroid hormone (PTH), prostaglandins, glucocorticoids and estrogen, have emerged as potential regulators of osteocyte function(s) and/or survival; furthermore, osteocytes synthesize and/or secrete proteins, such as FGF23, PHEX, MEPE and DMP1, that are of critical importance for the regulation of phosphorous homeostasis. As our understanding of these cells expands, it has become clear that they play a critical role in regulating and maintaining normal skeletal and mineral homeostasis. This Perspective reviews osteocyte function(s) and the role of osteocytes in bone remodeling and mineral homeostasis. IBMS BoneKEy. 2009 February;6(2):63-70. ©2009 International Bone & Mineral Society
Surfactant Protein A Prevents IFN-γ/IFN-γ Receptor Interaction and Attenuates Classical Activation of Human Alveolar Macrophages

Lung surfactant protein A (SP-A) plays an important function in modulating inflammation in the lung. However, the exact role of SP-A and the mechanism by which SP-A affects IFN-γ–induced activation of alveolar macrophages (aMϕs) remains unknown. To address these questions, we studied the effect of human SP-A on rat and human aMϕs stimulated with IFN-γ, LPS, and combinations thereof and measured the induction of proinflammatory mediators as well as SP-A’s ability to bind to IFN-γ or IFN-γR1. We found that SP-A inhibited (IFN-γ + LPS)–induced TNF-α, iNOS, and CXCL10 production by rat aMϕs. When rat macrophages were stimulated with LPS and IFN-γ separately, SP-A inhibited both LPS-induced signaling and IFN-γ–elicited STAT1 phosphorylation. SP-A also decreased TNF-α and CXCL10 secretion by ex vivo–cultured human aMϕs and M-CSF–derived macrophages stimulated by either LPS or IFN-γ or both. Hence, SP-A inhibited upregulation of IFN-γ–inducible genes (CXCL10, RARRES3, and ETV7) as well as STAT1 phosphorylation in human M-CSF–derived macrophages. In addition, we found that SP-A bound to human IFN-γ (KD = 11 ± 0.5 nM) in a Ca2+-dependent manner and prevented IFN-γ interaction with IFN-γR1 on human aMϕs. We conclude that SP-A inhibition of (IFN-γ + LPS) stimulation is due to SP-A attenuation of both inflammatory agents and that the binding of SP-A to IFN-γ abrogates IFN-γ effects on human macrophages, suppressing their classical activation and subsequent inflammatory response.