Miracle-dz/UniFAR-FTUs huggingface dataset:
Expression of apoptosis‐related genes in an Ethiopian cohort study correlates with tuberculosis clinical status

Mycobacterium tuberculosis remains one of the world's deadliest pathogens in part because of its ability to persist in the face of an active immune response. It has been suggested that apoptosis of infected macrophages is one way in which the host deals with intracellular pathogens and that M. tuberculosis can inhibit this process. To assess the relevance of this process for human disease, we compared the expression of multiple genes involved in the activation of the extrinsic (“death receptor initiated”) pathway of apoptosis in 29 tuberculosis patients, 70 tuberculosis contacts and 27 community controls from Ethiopia. We found that there is a strong upregulation of genes for factors that promote apoptosis in PBMC from individuals with active disease, including TNF‐α and its receptors, Fas and FasL and pro‐Caspase 8. The anti‐apoptotic factor FLIP, however, was also upregulated. A possible explanation for this dichotomy was given by fractionation of PBMC using CD14, which suggests that macrophage/monocytes may regulate several key molecules differently from non‐monocytic cells (especially TNF‐α and its receptors, a finding confirmed by protein ELISA) potentially reducing the sensitivity to apoptotic death of monocyte/macrophages – the primary host cell for M. tuberculosis. This may represent an important survival strategy for the pathogen.
Endometrial resection mandates reliable contraception thereafter – a case report of placenta increta following endometrial ablation

Placenta increta complicated pregnancy in a woman with a history of endometrial resection. Placentation in women with prior endometrial ablation carries a high risk for placenta accreta, increta and percreta. Contraceptive measures must be implemented after endometrial ablation and pursued until proven menopause, even in women who develop amenorrhoea postoperatively.
IL‐1: Discoveries, controversies and future directions

Although there has been a great amount of progress in the 25 years since the first reporting of the cDNA for IL‐1α and IL‐1β, the history of IL‐1 goes back to the early 1940s. In fact, the entire field of inflammatory cytokines, TLR and the innate immune response can be found in the story of IL‐1. This Viewpoint follows the steps from the identification of the fever‐inducing activities of “soluble factors” produced by endotoxin‐stimulated leukocytes through to the discovery of cryopyrin and the caspase‐1 inflammasome and on to the clinical benefits of anti‐IL‐1β‐based therapeutics. It also discusses some of the current controversies regarding the activation of the inflammasome. The future of novel anti‐inflammatory agents to combat chronic inflammation is based, in part, on the diseases that are uniquely responsive to anti‐IL‐1β, which is surely a reason to celebrate the 25th anniversary of the cloning of IL‐1α and IL‐1β.
OCORRÊNCIA DE Campylobacter jejuni EM LEITE CRU E PASTEURIZADO COMERCIALIZADO EM CURITIBA, ESTADO DO PARANÁ, BRASIL

Campylobacter jejuni is one of the most bacteria that cause food-borne diseases worldwide. Campylobacteriosis is commonly associated with the consumption of poultry meat; however, several outbreaks of the disease have also been linked to the ingestion of milk, usually due to failures in the pasteurization or even by raw milk’s consumption. This study aimed to evaluate the occurrence of C. jejuni in samples of raw or pasteurized milk, as well as evaluate different methodologies for analysis of this microorganism. Thirty milk samples were analyzed, 15 samples of raw milk and 15 samples of pasteurized milk, acquired in the metropolitan region of Curitiba, state of Parana, Brazil, between September 2012 and March 2013. The samples were subjected to counting of C. jejuni by direct plating and to the research after selective enrichment. None of the pasteurized milk samples showed C. jejuni contamination through both evaluated methods. By the direct plating method, were identified the contamination in two samples of raw milk, which presented counts of 4,0 x 10 1 and 1,65 x 10 2 CFU/mL. By using the selective enrichment method, C. jejuni was detected in four samples of raw milk, indicating the higher efficiency of the qualitative method in recovering injured cells and thus the higher reliability for detection of this pathogen. The occurrence of milk contaminated by C. jejuni should serve as a warning to guide preventive measures for the control and elimination of this pathogen in the production and manufacturing chain of the product.
Critical role of the calpain/calpastatin balance in acute allograft rejection

Rejection of solid organ allograft involves alloreactive T‐cell expansion. The importance of NF‐κB and NFAT in this process is underscored by the therapeutic efficacy of immunosuppressive agents, which target the two transcription factors. Since calpains, calcium‐activated proteases, are involved in the activation of NF‐κB and NFAT, we investigated the role of calpains in allograft rejection. In human transplant kidneys undergoing acute or chronic rejection, we show an increased expression of CAPN 1 gene encoding μ‐calpain, associated with a marked expression of μ‐calpain, mainly in infiltrating T cells. To address the role of calpain in rejection, we used a skin transplant model in transgenic mice expressing high levels of calpastatin, a calpain‐specific inhibitor. We show that calpain inhibition extended skin allograft survival, from 11 to 20 days. This delay was associated with a limitation in allograft infiltration by T cells. In vitro, calpain inhibition by calpastatin transgene expression limited dramatically T‐cell migration but, unexpectedly, increased slightly T‐cell proliferation. Amplification of IL‐2 signaling via the stabilization of IL‐2R common γ‐chain provided an explanation for the proliferation response. This is the first study establishing that calpain inhibition delays allograft rejection by slowing down T‐cell migration rather than proliferation.
Microbial succession in the rhizosphere of live and decomposing barley roots as affected by the antagonistic strain Pseudomonas fluorescens DR54-BN14 or the fungicide imazalil.

Abstract The protocol used in the present study was a long-term mesocosm experiment where the microbial succession around live barley roots and subsequent decomposing roots was assessed after seed coating with either the antagonistic strain Pseudomonas fluorescens DR54-BN14 or the fungicide imazalil. Four diversity measures were used: community level physiological profiles (CLPP), Bacteria-specific polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), actinomycete-specific PCR-DGGE and phospholipid fatty acid (PLFA), as well as total cell counts, colony-forming units (CFU) and culturable spore formers, and spore counts of the Bacillus cereus group. Analysis of non-treated plants provided a baseline description of the natural microbial succession from which effects of the treatments could be evaluated. A microbial succession occurred both in the rhizosphere and around decomposing roots, shown with all three diversity measures. A clear response to root death was found, and a clear distinction between root tip and root base samples. Using the recommended concentration of imazalil and a realistic number of DR54-BN14 for seed coating, transient, initial effects of both treatments on the microbial communities were observed at the root base with the PLFA analysis only. The lack of lasting significant side effects of DR54-BN14 is in agreement with an initial fast reduction in culturable DR54-BN14.
Differential requirement of IKK2 for CYLD‐dependent representation of thymic and peripheral T‐cell populations

The cylindromatosis tumor suppressor gene (Cyld) encodes an enzyme (CYLD) with deubiquitinating activity that has been implicated in the regulation of thymocyte selection in an NF‐κB‐essential‐modulator (NEMO)‐dependent manner. The main known molecular defects in thymocytes with inactive CYLD (LckCre‐Cyldflx9/flx9) are the aberrant hyperactivation of NF‐κB and JNK pathways. In order to dissect further the molecular mechanism of CYLD‐dependent thymocyte selection and address the role of NF‐κB specifically, we generated double mutant mice (LckCre‐Cyldflx9/flx9‐Ikk2flx/flx) in which CYLD was inactivated concomitantly with IKK2 (IκB‐kinase 2) in thymocytes. Interestingly, thymic development and NF‐κB activity in double mutant mice were fully restored, indicating that an IKK2‐dependent function of CYLD that leads to the hyperactivation of the NF‐κB pathway is primarily responsible for the defective selection of thymocytes. Intriguingly, we observed a greater reduction of CD4+ and CD8+ T cells in the periphery of LckCre‐Cyldflx9/flx9‐Ikk2flx/flx mice compared with LckCre‐Ikk2flx/flx mice. Collectively, our data establish CYLD as a critical regulator of thymocyte selection in a manner that depends on IKK2 and NF‐κB activation. In addition, our data uncover an IKK2‐independent role for CYLD in the establishment of physiological T‐cell populations in the periphery.
CXCR4 and CCR5 ligands cooperate in monocyte and lymphocyte migration and in inhibition of dual‐tropic (R5/X4) HIV‐1 infection

One of the most important functions of chemokines and their receptors is the regulation of directional migration of leukocytes within tissues. In specific tissue compartments, cells are exposed to multiple chemokines presented in complex dimensional and temporal patterns. Therefore, a leukocyte requires the mechanisms to integrate the various directional signals it receives from different chemoattractants. In this study, we report that CCL3, CCL5, and CCL8, three potent mononuclear cell chemoattractants, are able to synergize with the homeostatic chemokine CXCL12 in the migration of CD14+ monocytes, CD3+ T‐lymphocytes, or PHA‐activated lymphoblasts. In addition, CCL5 augmented the CXCR4 ligand‐driven ERK phosphorylation in mononuclear cells. Furthermore, the synergistic effect between CCL5 and CXCL12 in monocyte chemotaxis is inhibited in the presence of specific CCR1 antibody and AMD3100, but not by maraviroc. In HIV‐1 infection assays, a combination of CXCL12 and CCL5 cooperated to inhibit the replication of the dual‐tropic (R5/X4) HIV‐1 HE strain. Finally, although the dual‐tropic HIV‐1 strain was barely suppressed by AMD3100 or maraviroc alone, HIV‐1 infection was completely blocked by the combination of these two receptor antagonists. Our data demonstrate the cooperation between CCL5 and CXCL12, which has implications in migration of monocytes/lymphocytes during inflammation and in HIV‐1 infection.
Mesenchymal stem cell inhibition of T‐helper 17 cell‐ differentiation is triggered by cell–cell contact and mediated by prostaglandin E2 via the EP4 receptor

Mesenchymal stem cells (MSCs) inhibit T‐cell activation and proliferation but their effects on individual T‐cell‐effector pathways and on memory versus naïve T cells remain unclear. MSC influence on the differentiation of naïve and memory CD4+ T cells toward the Th17 phenotype was examined. CD4+ T cells exposed to Th17‐skewing conditions exhibited reduced CD25 and IL‐17A expression following MSC co‐culture. Inhibition of IL‐17A production persisted upon re‐stimulation in the absence of MSCs. These effects were attenuated when cell–cell contact was prevented. Th17 cultures from highly purified naïve‐ and memory‐phenotype responders were similarly inhibited. Th17 inhibition by MSCs was reversed by indomethacin and a selective COX‐2 inhibitor. Media from MSC/Th17 co‐cultures contained increased prostaglandin E2 (PGE2) levels and potently suppressed Th17 differentiation in fresh cultures. MSC‐mediated Th17 inhibition was reversed by a selective EP4 antagonist and was mimicked by synthetic PGE2 and a selective EP4 agonist. Activation‐induced IL‐17A secretion by naturally occurring, effector‐memory Th17 cells from a urinary obstruction model was also inhibited by MSC co‐culture in a COX‐dependent manner. Overall, MSCs potently inhibit Th17 differentiation from naïve and memory T‐cell precursors and inhibit naturally‐occurring Th17 cells derived from a site of inflammation. Suppression entails cell‐contact‐dependent COX‐2 induction resulting in direct Th17 inhibition by PGE2 via EP4.
The earliest intrathymic precursors of CD8α+ thymic dendritic cells correspond to myeloid‐type double‐negative 1c cells

The dendritic cells (DCs) present in lymphoid and non‐lymphoid organs are generated from progenitors with myeloid‐restricted potential. However, in the thymus a major subset of DCs expressing CD8α and langerin (CD207) appears to stand apart from all other DCs in that it is thought to derive from progenitors with lymphoid potential. Using mice expressing a fluorescent reporter and a diphtheria toxin receptor under the control of the cd207 gene, we demonstrated that CD207+CD8α+ thymic DCs do not share a common origin with T cells but originate from intrathymic precursors that express markers that are normally present on all (CD11c+ and MHCII molecules) or on some (CD207, CD135, CD8α, CX3CR1) DC subsets. Those intrathymic myeloid‐type precursors correspond to CD44+CD25– double‐negative 1c (DN1c) cells and are continuously renewed from bone marrow‐derived canonical DC precursors. In conclusion, our results demonstrate that the earliest intrathymic precursors of CD8α+ thymic DCs correspond to myeloid‐type DN1c cells and support the view that under physiological conditions myeloid‐restricted progenitors generate the whole constellation of DCs present in the body including the thymus.
Early B blasts acquire a capacity for Ig class switch recombination that is lost as they become plasmablasts

Rapid production of neutralizing antibody can be critical for limiting the spread of infection. Such early antibody results when B‐cell blasts mature directly to plasmablasts without forming germinal centers. These extrafollicular responses can involve Ig class switch recombination (CSR), producing antibody that can readily disseminate through infected tissues. The present study identifies the differentiation stage where CSR occurs in an extrafollicular response induced by 4‐hydroxy‐3‐nitrophenyl acetyl (NP) conjugated to Ficoll (NP‐Ficoll). To do this, we took advantage of the antigen dose dependency of CSR in this response. Thus, while both 30 and 1 μg NP‐Ficoll induce plasmablasts, only the higher antigen dose induces CSR. Activation‐induce cytidine deaminase (AID) is critical for CSR and in keeping with this a proportion of NP‐specific B‐cell blasts induced by 30 μg NP‐Ficoll express AID. None of the B blasts responding to the non‐CSR‐inducing 1 μg dose of NP‐Ficoll express AID. We confirmed that CSR occurs in B blasts by demonstrating the presence of rearranged heavy‐chain transcripts in B blasts in the 30 μg response. CSR in this extrafollicular response is confined to B blasts, because NP‐specific plasmablasts, identified by expressing CD138 and Blimp‐1, no longer express AID and cannot undergo CSR.
Prostaglandin E2 modulates IL‐8 expression through formation of a multiprotein enhanceosome in human colonic epithelial cells

Gastrointestinal inflammation is mediated by the pro‐inflammatory mediators interleukin‐8 (IL‐8) and prostaglandin E2 (PGE2). PGE2 binding and coupling through EP2/4 receptor subtypes on colonic epithelial cells stimulates cyclic adenosine monophosphate (cAMP) and IL‐8 production. Here we determined the mechanisms whereby PGE2 regu‐lates IL‐8 in Caco2 colonic epithelial cells and in cells over‐expressing the EP2/4 receptors (EP2S/EP4S). PGE2 coupling through EP2 activated the transcription factor inducible cAMP early repressor (ICER), whereas coupling through EP4 receptors activated the cyclic AMP‐responsive element‐binding protein (CREB). Activation of CREB in Caco2/EP2S was protein kinase A (PKA) dependent, whereas in EP4S cells, activation of CREB occurred through the PKA and phosphatidylinositol 3‐kinase pathways. Since ICER lacks the transactivation domain, it functions as a transcription repressor as opposed to CREB. PGE2 coupling through EP2/4 receptors can therefore acts in an opposing manner to either decrease (EP2) or promote IL‐8 expression by recruiting CREB‐binding protein (CBP) (EP4), which formed a multiprotein IL‐8 enhanceosome. A novel half CRE (167CRE) and a composite NFAT1‐AP1‐like site in the IL‐8 promoter participated in binding and complex formation as confirmed by mutagenesis and expression studies. These data unravel the mechanisms by which expression of IL‐8 is controlled by different signalling pathways that are activated by PGE2 but acting through different EP receptors.
The CD8+HLA‐DR+ T cells expanded in HIV‐1 infection are qualitatively identical to those from healthy controls

HIV‐induced immune activation leads to expansion of a subset of human CD8+ T cells expressing HLA‐DR antigens. Expansion of CD8+HLA‐DR+ T cells can be also observed in non‐HIV settings including several autoimmune diseases and aging. Although these cells are felt to represent “immune exhaustion” and/or to be anergic, their precise role in host defense has remained unclear. Here, we report that this subset of cells exhibits a restricted repertoire, shows evidence of multiple rounds of division, but lacks markers of recent TCR engagement. Detailed cell cycle analysis revealed that compared with their CD8+HLA‐DR− counterpart, the CD8+HLA‐DR+ T‐cell pool contained an increased fraction of cells in S‐phase with elevated levels of the G2/M regulators: cyclin A2, CDC25C, Cdc2 (CDK1), indicating that these cells are not truly anergic but rather experiencing proliferation in vivo. Together, these data support a hypothesis that antigen stimulation leads to the initial expansion of a CD8+ pool of cells in vivo that undergo further expansion independent of ongoing TCR engagement. No qualitative differences were noted between CD8+HLA‐DR+ cells from HIV+ and HIV− donors, indicating that the generation of CD8+HLA‐DR+ T cells is a part of normal immune regulation that is exaggerated in the setting of HIV‐1 infection.
Comparison of Healthy Life Style and Chronic Disease Management between Urban and Rural Older Adults

Purpose: The purpose of this study was to compare healthy life style and chronic disease management between urban and rural older adults. Methods: The study employed a comparative and descriptive survey design. Data were collected from 154 older adults living independently in communities (79 from urban and 75 from rural areas) using structured questionaries from 24 July, 2010 to 14 August, 2011. Results: Perceived health status was significantly lower in urban older adults than those in rural areas (
M‐CSF induces the expression of a membrane‐bound form of IL‐18 in a subset of human monocytes differentiating in vitro toward macrophages

IL‐18 is a proinflammatory cytokine belonging to the “IL‐1 family” that has been shown to play a prominent role in the induction of type 1 immune responses. Here, we show that M‐CSFinduces the expression of a membrane‐bound form of IL‐18 (mIL‐18) in a subset of human blood monocytes differentiating toward macrophages. While monocytes, DC, and GM‐CSF‐treated monocytes did not express mIL‐18, its expression was detected in approximately 30–40% of M‐CSF‐primed macrophages differentiating from both CD16− and CD16+ monocytes. Treatment with the caspase‐1 inhibitor significantly reduced mIL‐18 expression suggesting the requirement of an assembled inflammasome for IL‐18 surface expression. Polarization toward M2 did not modify mIL‐18 expression. On the contrary, LPS stimulation of both M0 and M2 (mIL‐18+) macrophages induced shedding of mIL‐18, which was likely mediated by the activation of cellular protease(s). Importantly, the soluble form IL‐18 (sIL‐18) induced in autologous resting NK cells both the expression of CCR7 and the production of high amounts of IFN‐γ, which was virtually abrogated by Ab‐mediated neutralization of sIL‐18. Overall our data shed new light on the cells and mechanisms leading to the release of sIL‐18, the major IFN‐γ‐inducing factor in both physiological and pathological immune responses.
Pediatric Hodgkin Lymphoma

Abstract We investigated the prognostic value of interim 18F-FDG PET/CT (PET-2) in pediatric Hodgkin lymphoma (pHL), evaluating both visual and semiquantitative analysis. Thirty pHL patients (age ⩽16) underwent serial 18F-FDG PET/CT: at baseline (PET-0), after 2 cycles of chemotherapy (PET-2) and at the end of first-line chemotherapy (PET-T). PET response assessment was carried out visually according to the Deauville Score (DS), as well as semiquantitatively by using the semiquantitative parameters reduction from PET-0 to PET-2 (&Dgr;&Sgr;SUVmax0–2, &Dgr;&Sgr;SUVmean0–2). Final clinical response assessment (outcome) at the end of first-line chemotherapy was the criterion standard, considering patients as responders (R) or nonresponders (NR). Disease status was followed identifying patients with absence or relapsed/progression disease (mean follow-up: 24 months, range 3–78). Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of visual and semiquantitative assessment were calculated; furthermore, Fisher exact test was performed to evaluate the association between both visual and semiquantitative assessment and outcome at the end of the first-line chemotherapy. The prognostic capability of PET-2 semiquantitative parameters was calculated by ROC analysis and expressed as area under curve (AUC). Finally, progression-free survival (PFS) was analyzed according to PET-2 results based on the 5-point scale and semiquantitative criteria, using the Kaplan–Meier method. Based on the outcome at the end of first-line chemotherapy, 5 of 30 patients were NR, the remnant 25 of 30 were R. Sensitivity, specificity, PPV, NPV, and accuracy of visual analysis were 60%,72%,30%,90%,70%; conversely, sensitivity, specificity, PPV, NPV, and accuracy of semiquantitative assessment were 80%, 92%, 66.7%, 95.8%, 90%. The highest AUC resulted for &Dgr;&Sgr;SUVmax0–2 (0.836; cut-off <12.5; sensitivity 80%; specificity 91%). The association between &Dgr;&Sgr;SUVmax0–2 and outcome at the end of first-line chemotherapy resulted to have a strong statistical significance (P = 0.0026). Both methods demonstrated to influence PFS, even if the semiquantitative assessment allowed a more accurate identification of patients with a high risk of treatment failure (P = 0.005). Our preliminary results showed that PET-2 visual assessment, by using Deauville criteria, can be improved by using the semiquantitative analysis. The SUV max reduction (&Dgr;&Sgr;SUVmax0–2) evaluation might provide a support for the interpretation of intermediate scores, predicting with good confidence those patients who will have a poor outcome and require alternative therapies.