[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.

Pertussis toxin inhibits activation-induced cell death of human thymocytes, pre-B leukemia cells and monocytes.

Activation of human thymocytes and pre-B cells via the CD3/T cell receptor (TCR) complex or the IgM/B cell receptor complex, respectively, results in apoptotic cell death. Similarly, cross-linking of the activation marker CD69, which belongs to the natural killer complex, causes apoptosis of lipopolysaccharide-preactivated monocytes. Here we show that pertussis toxin (PTX) inhibits the activation-induced apoptosis of these three cell types, though it fails to prevent the programmed cell death that follows exposure of cells to the synthetic glucocorticoid dexamethasone (thymocytes, pre-B cells) or to interleukin 4 (monocytes). The capacity of pertussis toxin to suppress activation-induced death is not due to quenching of the activation signal, because thymocytes exposed to PTX are still capable of mobilizing Ca2+ after TCR-alpha/beta cross-linking and proliferate in response to costimulation with PTX and CD3/TCR ligation. The apoptosis-inhibitory effect of PTX depends on the presence of an intact adenosine diphosphate (ADP)-ribosylating moiety, since a mutant pertussis toxin molecule that lacks enzymatic activity, but still possesses the membrane translocating activity, fails to interfere with activation-induced cell death. A toxin that induces a different spectrum of ADP ribosylation than PTX, cholera toxin, fails to inhibit apoptosis. To suppress apoptosis, the intact PTX holotoxin must be added to cells before the lethal activation step; its addition 30 min after initial activation remains without effect on apoptosis. These data unravel a PTX sensitive signal transduction event that intervenes during an early step of activation-induced cell death of immune cells.

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.

[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.

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.

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.

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.

Inhibition of apoptotic cell death in B-CLL by interferon gamma correlates with clinical stage.

Apoptosis was evaluated in B cells from 41 patients with B-CLL and 20 healthy aged-matched controls. B cells were cultured with and without gamma-IFN and other cytokines; apoptosis was quantified at regular intervals throughout a 5-day culture period. According to Rai's criteria, 17 patients were classified as good risk, 16 as intermediate and eight as high risk. In vitro, purified B cells from B-CLL patients were evaluated for apoptosis. Maximal apoptosis (44.12%) was observed at day 5 in cells from patients with poor prognosis. The addition of gamma-IFN to the culture media prevented apoptosis in a dose-dependent manner. Maximal inhibition of apoptosis was achieved with 100 IU/ml of gamma-IFN. The degree of inhibition of apoptosis by gamma-IFN was greater in cells from the high-risk group patients than in those from the intermediate and good prognosis group (P < 0.0001). The expression of gamma-IFN receptors in B-CLL cells was evaluated using a MnAb against the extracellular domain of gamma-IFN receptor. After 4 days in culture with gamma-IFN, only cells from the intermediate- and high-risk groups showed an increase in the density of gamma-IFN receptors (P < 0.001). gamma-IFN was not detected in the sera of our study patients. However gamma-IFN was detectable in the media from both normal B cells and B-CLL cells in culture; there was no difference in the amount of gamma-IFN released by cells from the three groups of patients studied. Our results show that in vivo gamma-IFN inhibits apoptosis of B cells from B-CLL patients. The inhibitory effect of gamma-IFN on apoptosis correlates directly with the severity of the disease and this is likely explained by a marked upregulation of gamma-IFN receptors in cells from patients in the high-risk group.

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.

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.

HLA class-II-mediated homotypic aggregation: involvement of a protein tyrosine kinase and protein kinase C.

Homotypic aggregation of B-lymphocytes, B-cell lines and class-II-positive T cells via HLA class II molecules was examined. Signaling via DR antigens induced rapid aggregation in a dose- and time-dependent manner, maximum and stable aggregation was induced within 20 minutes. On the contrary, rapid signaling via DP or DQ required prestimulation with either PMA or anti-sIg. Aggregation was temperature and energy dependent. [Ca2+] and [Mg2+] concentrations and an intact cytoskeleton were required while neither mRNA or protein synthesis were required. Furthermore, FACS analysis revealed that aggregation was not directly correlated with cell surface expression of HLA class II molecules. Our results demonstrate that aggregation was mediated through a protein tyrosine kinase (PTK)-dependent pathway that preceded activation of protein kinase C (PKC) and failure to generate either the PTK signal or the PKC signal prevented aggregation. The contribution of a tyrosine kinase was further demonstrated by the total inhibition of aggregation following treatment with an anti-CD45 mAb.

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.

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.

HLA class II-mediated aggregation is associated with the proliferation of B lymphocytes.

We have studied the role of LFA-1 antigens in human B lymphocyte aggregation, proliferation, and Ig production induced by a short stimulation via class II antigens. Cell stimulation with either bacterial superantigens or anti-DR mAbs rapidly induced homotypic cell aggregation. In response to IL-4, an increase in cell proliferation and Ig production was observed only when aggregation preceded addition of IL-4. The involvement of LFA-1 molecules in class II-induced aggregation was supported as LFA-1-deficient cells or B cells incubated with anti-LFA-1/ICAM-1 mAbs failed to aggregate after stimulation. The association between aggregation and subsequent Ig production and proliferation was further supported as, after IL-4 stimulation, in both LFA-1-deficient cells and B cells incubated with anti-LFA-1 mAbs, class II-mediated signals failed to increase Ig production or cell proliferation. These data suggest that in class II-stimulated cells, LFA-1-dependent aggregation has a major role in IL-4-dependent Ig production and proliferation of B cells.

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.