[Analysis of pediatric «short stays¼ in a second-level hospital throughout 25 years]. / Análisis de las «estancias cero¼ pediátricas en un hospital de nivel 2 a lo largo de 25 años.

INTRODUCTION: It is essential to admit patients to hospital in an efficient way in order to use resources rationally. Short hospitalary stays are hospitalizations which does not include 00:00h and are considered avoidable. This study describes trends and characteristics of short stays throughout 25 years in our hospital. PATIENTS AND METHODS: We analyzed hospital pediatric discharges in a second-level hospital through the registration system «conjunto mínimo básico de datos¼. We categorized pediatric patients and newborn patients in two groups according to length of hospital stay: «short stays¼ and «prolonged stays¼. We analyzed and compared the following variables: gender, age, type of admission, month, diagnosis-related groups (DRG) and admission service. Binary logistic regression analysis and assessment of trends through joinpoint regression analysis were performed. RESULTS: From 1993 to 2017, 45710 children were admitted to our hospital, of which 7.3% were short stays. The trend analysis showed a point of change upwards-downwards at the beginning of the millennium. Pediatric short stays: the most important variables were emergency admissions (89%), urgent transfers (9%), month December (11%) and main diagnosis category: nervous system (18%). Mean diagnosis-related groups cost was 2432±1115€ in short stays group and 2549±1065€ in prolonged stays. CONCLUSIONS: Short stays and prolonged stays show a falling trend in our hospital. Short stays percentage in our environment is similar to other neighbor countries. Some of our short stays are urgent transfers and admissions for clinical observation. We did not find clinical significance in weight or cost of pediatric patients' DRG comparing to prolonged stays.

Microvesicles: ROS scavengers and ROS producers.

This review analyzes the relationship between microvesicles and reactive oxygen species (ROS). This relationship is bidirectional; on the one hand, the number and content of microvesicles produced by the cells are affected by oxidative stress conditions; on the other hand, microvesicles can directly and/or indirectly modify the ROS content in the extra- as well as the intracellular compartments. In this regard, microvesicles contain a pro-oxidant or antioxidant machinery that may produce or scavenge ROS: direct effect. This mechanism is especially suitable for eliminating ROS in the extracellular compartment. Endothelial microvesicles, in particular, contain a specific and well-developed antioxidant machinery. On the other hand, the molecules included in microvesicles can modify (activate or inhibit) ROS metabolism in their target cells: indirect effect. This can be achieved by the incorporation into the cells of ROS metabolic enzymes included in the microvesicles, or by the regulation of signaling pathways involved in ROS metabolism. Proteins, as well as miRNAs, are involved in this last effect.

Donor-Specific Antibodies After Starting Hemodialysis in Nonrenal Solid Organ Transplant Recipients: Role of TH17.

BACKGROUND: Nonrenal transplantation could cause a progressive deterioration in renal function until need dialysis. It is important to know if these patients increased their risk to develop de novo donor-specific anti-HLA antibody (DSA) after starting hemodialysis (HD) and if so, try to find the mechanism. MATERIAL AND METHODS: In this double-phase study, we first analyzed the incidence of development DSA in nonrenal transplant recipients after starting HD by a retrospective study. Secondly, a prospective study was designed to analyze the pharmacokinetics of immunosuppressive drugs and the cytokine profile of these patients. RESULTS: Of 179 pancreas transplant recipients, 16 needed to start HD, and 62.5% of these patients developed de novo DSA after starting HD, with 80% of them class I DSA. In the second phase of the study, the plasma levels of the immunosuppressive drugs as measured by a limited sampling strategy of 3 sample time points (C0, C2, and C4) were stable. The cytokine profile showed that there was an increase in Th1 cytokine (interferon gamma of 0.045 ng/mL) and also in Th17 cytokines (transforming growth factor ß >10 ng/mL). CONCLUSION: Our data suggest that the development of DSA after starting HD in nonrenal transplant recipients could be mediated by Th17 immune response mechanisms.

Klotho Prevents Translocation of NFκB.

Klotho protein is a ß-glucuronidase capable of hydrolyzing steroid ß-glucuronides. Two molecules are produced by the Klotho gene, a membrane bound form and a circulating form. This protein is recognized as an antiaging gene with pleiotropic functions. The activation of cellular systems is associated with the pathogenesis of several chronic and degenerative diseases associated with an inflammatory state. Inflammation is characterized by an activation of NFκB. Klotho suppresses nuclear factor NFκB activation and the subsequent transcription of proinflammatory genes. This review focuses on the current understanding of Klotho protein function and its relationship with NFκB regulation, emphasizing its potential involvement in the pathophysiologic process.

Home e-health system integration in the Smart Home through a common media server.

Home e-health systems and services are revealed as one of the most important challenges to promote Quality of Life related to Health in the Information Society. Leading companies have worked on e-health systems although the majority of them are addressed to hospital or primary care settings. The solution detailed in this paper offers a personal health system to be integrated with Smart Home services platform to support home based e-care. Thus, the home e-health system and architecture detailed in this research work is ready to supply a seamless personal care solution both from the biomedical data analysis, service provision, security guarantee and information management s point of view. The solution is ready to be integrated within the Accessible Digital Home, a living lab managed by Universidad Politécnica de Madrid for R&D activities.

Microinflammation induces endothelial damage in hemodialysis patients: the role of convective transport.

Cardiovascular complications are a major cause of mortality in hemodialysis patients. On-line hemofiltration combines convective clearance for removing large solutes with diffusion to remove small solutes and is associated with a significant reduction of inflammation and improved patient survival. We compared on-line hemofiltration to high-flux hemodialysis (HF-HD) in patients in a sequential manner. At baseline, 15 stable patients on HF-HD as compared with five control subjects showed significant increases in CD14+CD16+ cells, endothelial microparticles, and endothelial progenitor cells (EPCs). After 4 months of on-line hemofiltration, the number of CD14+CD16+ cells, microparticles, and EPCs decreased. After returning to HF-HD for 4 months, all measured parameters returned to their respective baseline values. The number of CD14+CD16+ cells correlated with both endothelial microparticles and EPCs. We conclude that on-line hemofiltration attenuates endothelial dysfunction possibly by decreasing microinflammation. This effect may be directly caused by a modulatory effect of on-line hemofiltration on proinflammatory cells or by a complex interaction that encompasses a wider removal of uremic toxins.

[Acute rejection of kidney transplant is associated with induction of replicative senescence in CD8+ T lymphocytes]. / Senescencia replicativa en células T CD8+ de pacientes con rechazo agudo en trasplante renal.

Acute renal rejection repeatedly activates immunocompromised CD8 + T cells. Maintained activation of CD8 + T cells can induce a process of replicative senescence. In the present study, we will evaluate in CD8 lymphocytes from patients undergoing acute renal rejection characteristics of replicative senescence such as: a) low expression of CD28 molecule; b) telomere shortening and c) increase production of proinflammatory cytokines. The study was carried out in CD8 + T cells from 14 patients transplanted without clinical evidences of acute renal rejection, 14 patients kidney transplanted with clinical and anatomopathological evidences of acute renal rejection, 8 healthy controls. The results shown that in peripheral blood and renal biopsy of patients with acute renal rejection there is a significant increment of the population of T cells CD28-CD8+, with short telomere length, as compared with healthy controls and patients without acute renal rejection. The presence of senescent cells was associated with high levels of IL-10 and IFN-Y in plasma and urine. In conclusion our study suggest that the CD8 + T cells of patients with acute renal rejection suffer a process of replicative senescence.

Senescencia replicativa en células T CD8+ de pacientes con rechazo agudo en trasplante renal / Acute renal rejection is associated with induction of replicative senescence in CD8+ T lymphocytes

Durante el rechazo renal agudo las células T CD8+ son activadas de forma mantenida pudiendo desencadenar un proceso de senescencia replicativa. En este trabajo nosotros hemos evaluado varias características de senescencia replicativa como es : a) porcentaje de células T CD28-; b) acortamiento telomérico, y c) producción de citoquinas proinflamatorias. Este estudio se realizó en 14 pacientes con rechazo renal agudo, 14 pacientes sin rechazo renal agudo y 8 controles sanos. Los resultados indicaron que los pacientes con rechazo renal agudo mostraban altos porcentajes de células T CD28-CD8+ en sangre periférica y biopsia renal. Estos pacientes mostraron telómeros cortos que se asociaban con el fenotipo CD28-. Además, los pacientes con rechazo renal agudo presentaban altas concentraciones de IL-10 e IFN-beta; en plasma y orina. Nuestro estudio demuestra que las células T CD8+ de pacientes con rechazo renal agudo sufren un proceso de senescencia replicativa

Acute renal rejection repeatedly activates immunocompromised CD8 + T cells. Maintained activation of CD8 + T cells can induce a process of replicative senescence. In the present study, we will evaluate in CD8 lymphocytes from patients undergoing acute renal rejection characteristics of replicative senescence such as: a) low expression of CD28 molecule; b) telomere shortening and c) increase production of proinflammatory cytokines. The study was carried out in CD8 + T cells from 14 patients transplanted without clinical evidences of acute renal rejection, 14 patients kidney transplanted with clinical and anatomopathological evidences of acute renal rejection, 8 healthy controls. The results shown that in peripheral blood and renal biopsy of patients with acute renal rejection there is a significant increment of the population of T cells CD28-CD8+, with short telomere length, as compared with healthy controls and patients without acute renal rejection. The presence of senescent cells was associated with high levels of IL-10 and IFN-beta; in plasma and urine. In conclusion our study suggest that the CD8 + T cells of patients with acute renal rejection suffer a process of replicative senescence

Antibody-mediated pure red-cell aplasia (PRCA): the Spanish experience.

BACKGROUND: The incidence of antibody (Ab)-mediated pure red-cell aplasia (PRCA) in patients with chronic kidney disease (CKD) has increased between 1998 and 2002. After initially responding to treatment with recombinant human erythropoietic agents for CKD-associated anemia, patients became treatment-refractory and severely anemic. Although most PRCA cases have occurred in Europe, the varying epidemiologies among individual countries have not been well characterized. METHODS: We investigated Ab-mediated PRCA in 12 Spanish patients treated with epoetin alfa alone or prior to treatment with epoetin beta (n=1) or darbepoetin alfa (n=1). Serum Abs against epoetin alfa were detected by radioimmunoprecipitation (RIP) assay or bioassay. Following diagnosis of PRCA, erythropoietic treatment was stopped and patients received immunosuppressive therapy alone (n=11) or in combination with renal transplant (n=1). RESULTS: Treatments were administered for 16 months (average) before diagnosis of PRCA in bone marrow aspirates (n=8) or biopsies (n=4). At diagnosis, patients had an average of 0.68% blood reticulocytes and blood hemoglobin (Hb) level of 7.13 g/dL. Eight patients had anti-epoetin Abs detected by RIP, and 5 had neutralizing Abs measured in the bioassay. As of December 2003, 4 patients had died, 3 had no recovery, and 5 had recovered from anemia (blood Hb level, 9.9 g/dL). All 5 recovering patients received corticosteroid therapy alone, and 1 received a renal transplant as well as corticosteroids. CONCLUSIONS: Sudden onset of treatment-refractory anemia in CKD patients suggests a course of treatment cessation followed by diagnostic procedures for Ab-mediated PRCA, and immunosuppressive therapy. This study may serve as a model for a centralized global PRCA registry.

Antithrombin III prevents early pulmonary dysfunction after lung transplantation in the dog.

BACKGROUND: Ischemia-reperfusion injury with the resulting inflammatory response is a devastating complication of lung transplantation; much of the tissue damage could be diminished by control of the inflammatory response. Recent studies have show that antithrombin III (AT III) has an anti-inflammatory effect in addition to its established role in the regulation of blood coagulation. Thus, we hypothesized that the administration of AT III might help to prevent ischemia-reperfusion injury after lung transplantation. METHODS AND RESULTS: The study was performed in a dog model of orthotopic lung transplantation. Dogs were randomly assigned to receive either vehicle (controls) or AT III. We observed that in control dogs, during the 180-minute period after lung transplantation, the arterial O(2) partial pressure decreased and both the alveolar-arterial O(2) difference and the pulmonary vascular resistance increased. By contrast, these parameters remained unchanged in the group of dogs receiving AT III. Dogs with transplants receiving AT III did not show an increase in cell adhesion molecules, and histological examination revealed almost an absence of inflammatory response. The administration of AT III produced a marked increase in serum prostacyclin (PGI(2)) levels, whereas in control dogs, the PGI(2) levels did not change. The beneficial effect of AT III was not observed when dogs received indomethacin to prevent the stimulation of PGI(2) release by AT III. CONCLUSIONS: Our results demonstrate that AT III prevents ischemia-reperfusion injury in a dog model of lung transplantation and that this effect is conditioned by an increase in PGI(2) production.

Role of adhesion molecules in mononuclear cell apoptosis induced by cuprophan hemodialysis membranes.

BACKGROUND/AIM: Hemodialysis with Cuprophan (CU) membranes induces mononuclear cell activation, leading to increased expression of adhesion molecules, formation of cell aggregates, and apoptosis. It is likely that structure(s) of the CU membrane interact with mononuclear cell surface molecules which transduce biochemical signals to the cell. Interactions between adhesion molecules and extracellular matrix have been implicated in cell activation, proliferation, and/or apoptosis. In the present work, we study whether adhesion molecules may be involved in CU-induced mononuclear cell aggregation and/or apoptosis. METHODS: The present study was performed using THP-1 cells, a human monocytic cell line, cultured in the presence of the CU membrane. CD11b and CD54 expression was studied with fluorescent monoclonal antibodies. Cell aggregation was quantified using a phase-contrast microscope. Apoptosis was evaluated by either light microscopy or annexin V labeling. RESULTS: The results show that incubation of CU membranes with the proteins CD11b, CD18, and CD54 or the blockade of these cell surface molecules with specific monoclonal antibodies inhibited the CU-induced aggregation and apoptosis in a dose-dependent manner. CONCLUSION: These results suggest that CU membranes interact selectively with these specific proteins to induce cell activation which ultimately results in apoptosis.

TH2 lymphocytes from atopic patients treated with immunotherapy undergo rapid apoptosis after culture with specific allergens.

BACKGROUND: In atopic patients treatment with specific immunotherapy (SIT) induced a shift in the balance of T-cell immune response away from a T(H)2-type (producing mostly IL-4) in favor of a T(H)1-type T-lymphocyte response (with the preferential production of IFN-gamma). However, the mechanisms through which SIT acts are less clear. We have recently shown that allergens may induce an activation-induced cell death process in lymphocytes from SIT-treated atopic patients. OBJECTIVE: This study aimed to determine whether allergen-induced apoptosis can occur in a specific subset of cells. METHODS: The study was performed in lymphocytes from normal subjects and atopic patients, some of whom were treated with SIT. Cells were cultured in the presence of gramineous pollen (Lolium perenne) allergenic extracts. Cell phenotype and intracellular cytokine expression were measured by means of fluorescent mAbs. Apoptosis was measured by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling. Fluorescence was analyzed in a FACScan flow cytometer, and the data were evaluated with Consort 30 software. RESULTS: Our results showed that allergens induce apoptosis of lymphocytes in SIT-treated atopic patients. Apoptosis occurs mainly in T(H)2 lymphocytes with the IL-4+/CD4+ phenotype and subsequently increases the percentage of IFN-gamma(+) cells in the culture. CONCLUSION: These results suggest that the shift from T(H)2 to T(H)1 induced by SIT in atopic patients may be mediated, at least in part, by the induction of an activation-induced cell death process in allergen-responder T(H)2 cells.

Allergens induce apoptosis in lymphocytes from atopic patients.

Repeated stimulation of immune cells may induce an "activation-induced cell death" (AICD) program. Allergy is characterized by the cyclic activation of allergen-reactive immune cells. To study the effects of allergen stimulation in cell proliferation and apoptosis in atopic subjects, peripheral blood mononuclear cells (PBL) from 40 atopic patients with positive reactivity to the allergens Olea Europaea (OE) and Lollium Perenne (LP) (20 without immunotherapy and 20 with specific immunotherapy) and 10 normal subjects were cultured with the allergens OE and LP. PBL from atopic patients proliferate more vigorously than cells from normal subjects after culture in vitro with both allergens, although PBL from atopic subjects without immunotherapy proliferate more than PBL from atopic subjects with immunotherapy. The study of cell proliferation shows that in atopic patients PBL mainly exhibit the CD4/CD45RO phenotype. This preferential proliferation is more evident in PBL from atopic patients treated without immunotherapy. Cell culture with specific allergens induces apoptosis in PBL from atopic patients. The percentage of apoptosis increased when atopic patients had been previously treated with immunotherapy. In addition to the observed increase in cell proliferation, apoptosis mainly occurs in the CD45RO cells that support the involvement of these cells in allergy. Furthermore, results obtained in cells from immunized patients suggest that an AICD process may partly at least explain the mechanism of action of allergen immunotherapy.

NK phenotypic markers and IL2 response in NK cells from elderly people.

Immunosenescence is a process that primarily affects the T cell compartment of the immune system, although age-associated immunological alterations have also been demonstrated in the NK cell phenotype and function. A significant expansion in the number of NK cells is found in aging. The NK cytotoxic capacity of total peripheral blood lymphocytes is also well preserved, not only in healthy elderly people but also in centenarians. However, NK cell killing of K562 is impaired when considered in a per-cell basis, and this defect is associated with defective signal transduction after activation more than a diminished conjugate formation or killing capacity. We have studied the phenotype of NK cells in elderly donors fulfilling the Senieur criteria. We have also studied the capacity of these cells to be activated by IL2 when different NK cell functions, other than cytotoxicity, are considered. Our results confirm the increased percentage of NK cells in the elderly due to the expansion of the CD56dim subset that also show an altered pattern of activation markers, whereas no differences were found in the CD56bright subset. The response of NK cells to IL2 was found to be impaired when proliferation, expression of CD69, and Ca2+ mobilization were considered, whereas TNF-alpha production was not significantly affected. These results suggest that human NK cells do not escape the aging process, although senescence have a differential effect on distinct NK cell biological functions, ranging from severe to negligible impairment, depending on the parameters considered.

Serum levels of intercellular adhesion molecule 1 (ICAM-1) in patients with colorectal cancer: inhibitory effect on cytotoxicity.

A positive correlation between the level of ICAM-1 in serum and the stage of neoplastic processes has been demonstrated. We studied ICAM-1 serum concentration in 27 colorectal cancer patients and investigated the effect of this molecule on cellular aggregation and toxicity. ICAM-1 serum concentration in the group of patients was significantly higher (P < 0.01) than in normal controls and was related to tumour stage. Patient sera inhibited both the formation of cellular aggregates and the percentage of specific lysis, the effect being lost when the serum was depleted of ICAM-1. These results suggest that the release of soluble ICAM-1 may represent a mechanism of tumour escape.

Nonbiocompatible hemodialysis membranes induce apoptosis in mononuclear cells: the role of G-proteins.

Mononuclear cells are activated during hemodialysis. In this study, we provide evidence that in vitro culture of mononuclear cells with Cuprophan, a nonbiocompatible hemodialysis membrane, increases the levels of protein phosphorylation in these cells as well as the expression of surface activation molecules. By contrast, culturing of mononuclear cells with AN69, a more biocompatible membrane, did not increase protein phosphorylation levels or expression of surface activation molecules in these cells. In addition, Cuprophan, but not the AN69 membrane, increased the percentage of mononuclear cell death by apoptosis. Inhibition of G-protein-mediated signal transduction decreased the apoptosis of cells cultured with the Cuprophan membrane. The GTP-binding protein involved in Cuprophan-induced apoptosis was sensitive to the ADP-ribosylating pertussis toxin (PTX). The inhibition of GTP-binding protein decreased apoptosis in the early stage of the activation-induced apoptosis, suggesting that G-proteins are implicated in the transmission of apoptosis-inducing signal(s) but do not interfere with the effector signals that mediate the late stages of apoptotic catabolism. Finally, PTX was capable of inhibiting apoptosis without affecting expression of activation molecules; thus, the inhibition of apoptosis by Cuprophan was not due to quenching of the stimulation signals, because monocytes were still able to be activated by Cuprophan despite the action of PTX. The results obtained in this study suggest that cell activation and apoptosis may be mediated by separate intracellular signals.

C1-esterase inhibitor prevents early pulmonary dysfunction after lung transplantation in the dog.

The success of lung transplantation to a large extent depends on effective protection of the graft from ischemic injury after reperfusion. Although mechanisms have not been clarified, the pathologic findings of ischemic injury after reperfusion are similar to adult respiratory distress syndrome, a condition in which the blood coagulation contact system is activated. This study evaluates the effect of C1-esterase inhibitor (C1-INH), the main inhibitor of the blood coagulation contact system, on short-term lung function in a dog model of orthotopic lung transplantation. Twelve lung transplantations were performed after 24 h of ischemic time. Dogs were randomly assigned to receive either vehicle (Control) or C1-INH. After the lung transplantation in the control group, Pao2 decreased by 84% and both the AaPO2 and the Qs/Qt% increased (340 and 530%, respectively, p < 0.01); these parameters remained unchanged in the C1-INH group. The hypoxemia observed in control animals was associated with decreased blood coagulation contact factors, complement consumption, increased expression of adhesion glycoproteins in leukocytes, and extensive intraalveolar and interstitial neutrophil infiltration. In contrast, C1-INH administration prevented hypoxemia, the decrease in blood coagulation contact factors, the activation of the complement system, the increase in expression of leukocyte adhesion molecules, and inflammatory cell infiltrate. This study has demonstrated that in a dog model of lung transplantation, the administration of C1-INH prevents early pulmonary dysfunction, and it suggests that activation of blood coagulation contact system and complement are important mechanisms causing ischemic injury after reperfusion.

Mycobacterium tuberculosis induces apoptosis in gamma/delta T lymphocytes from patients with advanced clinical forms of active tuberculosis.

Antigens from inactivated Mycobacterium tuberculosis H37Ra induce activation in a subpopulation of gamma/delta (gamma/delta) T lymphocytes in a manner that resembles that of superantigens from alpha/beta T cells. After culture in vitro with H37Ra proteins, gamma/delta T lymphocytes from patients with advanced clinical forms of active tuberculosis (ACF-TBC) display cytotoxic activity against homotypic target cells exposed to H37Ra. Cytotoxicity by gamma/delta T lymphocytes from ACF-TBC patients occurs in a range similar to that observed in healthy subjects. Following activation, H37Ra-stimulated gamma/delta T lymphocytes from healthy subjects did proliferate in the presence of exogenous recombinant human interleukin 2. However, under the same conditions, gamma/delta T lymphocytes from ACF-TBC patients not only did not proliferate but died by apoptosis. These results suggest that in gamma/delta T lymphocytes from patients with ACF-TBC, antigens from M. tuberculosis may induce cell activation that leads to apoptotic cell death.