Gene expression, protein profiling, and chemotactic activity of infrapatellar fat pad mesenchymal stem cells in pathologies of the knee joint.

The infrapatellar fat pad (IPFP) is a periarticular adipose knee tissue. This tissue contains a large number of mesenchymal stem cells (MSCs). In the present work, we wanted to study the IPFP MSCs and their relationship and differences in two groups, anterior cruciate ligament (ACL) ruptures knees and ostheoarthrosis (OA). The IPFP of 42 patients with OA or ACL rupture were analyzed. Isolation, primary culture, and a genetic and proteomic study of MSCs from IPFP were performed. Gene expression of IL-6, tumor necrosis factor (TNF), IL-8, HSPA1A (Hsp70), CXCL10, RANTES, MMP1, MMP3, TIMP1, and BMP7 was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). We analyzed MSCs from from 12 diferents patients in two cellular pools (6 from AO disease and 6 from ALC rupture to form two cell pool), for the iTRAQ Proteomic Assay. The conditional media were used in quantitative analysis of MSC soluble factors by Luminex and for de migration assay. A higher gene expression of IL-6, TNF, CXCL10, RANTES, and MMP1 and OPG in MSCs from OA versus ACL (p < 0.05) was observed. Conversely HSPA1A, TIMP1, and RANKL showed a significant lower expression in OA-MSCs (p < 0.05). In the secretome analysis, adipsin and visfantin levels in the supernatants from OA-MSCs were lower (p < 0.05) respect to ACL-MSCs. Also, the monocytic cells migrated two-folds in the presence of conditioned media from OA-MSCs patients versus patients with ACL-MSC. The infrapatellar pad should be considered as an adipose tissue capable of producing and excreting inflammatory mediators directly in the knee joint, influencing the development and progression of knee joint pathologies.

Human decidual stromal cells express HLA-G: Effects of cytokines and decidualization.

BACKGROUND: Decidual stromal cells (DSC) are the main cellular component of the decidua, the maternal tissue in close contact with fetal trophoblast. Although of mesenchymal origin, DSC exert numerous immune functions that seem to be relevant for the immunological relationship between the mother and fetus. HLA-G, an antigen preferentially expressed by trophoblast, appears to participate in the immune tolerance by the mother of the semiallogeneic fetus. METHODS AND RESULTS: We show by flow cytometry, fluorescence microscopy, western blotting and RT-PCR that DSC isolated and maintained in culture express HLA-G weakly but consistently. We also detected this antigen by flow cytometry in fresh DSC. Interleukin (IL)-10, a cytokine associated with normal pregnancy, increased the expression of HLA-G by DSC (P < 0.00001), whereas IL-2, a cytokine involved in spontaneous abortion, showed no effect. Decidualization by progesterone and cAMP also up-regulated the expression of HLA-G by DSC (P < 0.001). Interferon gamma, a cytokine implicated in the vascular remodelling of the decidua necessary for embryo implantation, also increased the expression of HLA-G by DSC (P < 0.05). CONCLUSIONS: Our results suggest the existence of a network in which hormones together with cytokines regulate the expression of HLA-G by DSC, and that may be of relevance in the maintenance of maternal-fetal tolerance.

Contractile activity of human decidual stromal cells. II. Effect of interleukin-10.

CONTEXT: Human decidual stromal cells (DSC) are myofibroblast-like cells that express alpha-smooth muscle (alpha-SM) actin, a protein associated with cell contractility. Several lines of experimental evidence in humans and mice show that antiinflammatory cytokines favor normal pregnancy, whereas Th1 and inflammatory cytokines play a role in abortion. We previously demonstrated that IL-2, a Th1 cytokine, increased the contractility of human DSC. OBJECTIVE: We studied the effect of the antiinflammatory cytokines IL-10 and IL-4 on the contractility of DSC from first-trimester pregnancy. SETTING AND PATIENTS: We studied 10 healthy women who underwent elective vaginal termination of first-trimester pregnancy at Clínica El Sur, Málaga, and Clínica Ginegranada, Granada. MAIN OUTCOME MEASURE(S): After isolation of DSC, cell contractility was measured with the collagen gel contraction assay. alpha-SM actin was detected with Western blotting and immunofluorescence. RESULTS: We found that IL-10, but not IL-4, increased the volume of the collagen gel matrixes in which the cytokine-treated DSC were cultured, showing that IL-10 decreased DSC contractility. By Western blotting we demonstrated that this effect was not related to an alteration in the synthesis of alpha-SM actin. Nevertheless, we observed by immunofluorescence microscopy that DSC treated with IL-10 exhibited stress fibers with a lower content of alpha-SM actin than untreated control DSC. CONCLUSIONS: IL-10 relaxes DSC by reducing the incorporation of alpha-SM actin into their stress fibers. This relaxing activity may be of relevance for the maintenance of pregnancy.

A pivotal role for galectin-1 in fetomaternal tolerance.

A successful pregnancy requires synchronized adaptation of maternal immune-endocrine mechanisms to the fetus. Here we show that galectin-1 (Gal-1), an immunoregulatory glycan-binding protein, has a pivotal role in conferring fetomaternal tolerance. Consistently with a marked decrease in Gal-1 expression during failing pregnancies, Gal-1-deficient (Lgals1-/-) mice showed higher rates of fetal loss compared to wild-type mice in allogeneic matings, whereas fetal survival was unaffected in syngeneic matings. Treatment with recombinant Gal-1 prevented fetal loss and restored tolerance through multiple mechanisms, including the induction of tolerogenic dendritic cells, which in turn promoted the expansion of interleukin-10 (IL-10)-secreting regulatory T cells in vivo. Accordingly, Gal-1's protective effects were abrogated in mice depleted of regulatory T cells or deficient in IL-10. In addition, we provide evidence for synergy between Gal-1 and progesterone in the maintenance of pregnancy. Thus, Gal-1 is a pivotal regulator of fetomaternal tolerance that has potential therapeutic implications in threatened pregnancies.

Valor pronóstico de las alteraciones plaquetarias en meningoencefalitis bacteriana / Pronostics valor of the changes platelet in bacteria meningoencephalitis

Este estudio prospectivo fue realizado para conocer la relación pronóstica de las modificaciones plaquetarias en pacientes con meningoencefalitis. Se estudiaron sesenta y tres (63) pacientes menores de 13 años con diagnóstico de meningoencefalitis bacteriana, con seguimiento de las cifras de plaquetas durante su hospitalización (al ingreso, al quinto y al octavo día). En los resultados obtenidos hubo correlación de trombocitosis con el buen pronóstico y la trombocitopenia con pronósticos contrarios. Por tanto, es recomendable tomar en cuenta la determinación de los valores de cifras plaquetarias en dichos pacientes por su utilidad pronóstica

Tratamiento empiríco del absceso hepático pedíatrico en Venezuela: experiencia clínica con 54 casos en menores de 13 años / Empirical therapy of the amibic liver abscess in pediatric in Venezuela

Se revisa en forma retrospectiva la experiencia obtenida durante 9 años consecutivos con 54 niños en quienes se confirmó el diagnóstico de absceso hepático. El proceso predominó en el grupo etario de 2-6 años, la duración promedio previa a la hospitalización fue de 7 días o más, en 26 por ciento de los pacientes existían antecedentes disentéricos y en 19 por ciento de amibiasis intestinal. Fiebre (94 por ciento), hepatomegalia (93 por ciento), dolor abdominal (83 por ciento) y anormalidades a la auscultación en la base pulmonar derecha (54 por ciento), fueron los hallazgos clínicos predominantes. Eritrosedimentación elevada (94 por ciento), leucocitosis con neutrofilia (89 por ciento), anemia (87 por ciento), prolongación del tiempo de protrombina (83 por ciento), hipoalbuminemia (80 por ciento) y elevación de las transaminasas séricas (47 por ciento) fueron las anormalidades de laboratotio más frecuentes. las lesiones ecográficas fueron únicas en 69 por ciento de los casos y predominaron en el lóbulo derecho (67 por ciento), especialmente en su segmento posterior (49 por ciento). el diámetro de las lesiones fue >5 cm en 76 por ciento de los casos. se requirió drenaje quirúrgico coadyuvante en 28 por ciento de los pacientes y la estancia hospitalaria fue mayor de 21 días en 78 por ciento de los casos. La tasa de letalidad fue 7,4 por ciento correspondiente uno de los 4 casos mortales a un absceso de etiología escaridiana