Generation, establishment and characterization of a pluripotent stem cell line (CVTTHi001-A) from primary fibroblasts isolated from a patient with activated PI3 kinase delta syndrome (APDS2).

APDS2 is caused by mutations in PIK3R1 gene resulting in constitutive PI3Kδ activation. PI3Kδ is predominantly expressed in leukocytes and plays critical roles in regulating immune responses. Here we first derived fibroblast primary cells from a skin biopsy of a patient carrying a heterozygous single T deletion in intron 11 of the PIK3R1 gene. We next present the derivation of an induced pluripotent stem cell (iPS) line using a non-integrative reprogramming technology. Pluripotent-related hallmarks are further shown, including: iPSCs self-renewal and expression of pluripotent and differentiation markers after in vitro differentiation towards embryonic germ layers, assessed by RT-PCR and immunofluorescence.

El colecho: de los discursos de moda al psicoanálisis / Co-sleeping: from current trends to psychoanalysis

Con el objetivo de contrastar los postulados de la "Crianza con Apego" propuesta por William Sears, sustentada en la "Teoría del Apego" desarrollada por John Bowlby y reivindicada por diversos autores contemporáneos que defienden la práctica del colecho en la actualidad, analizaré los conceptos del psicoanálisis que se pronuncian en contra de la misma a través de una revisión bibliográfica exploratoria que busca profundizar los conocimientos sobre el colecho.

With the aim of contrasting the premises of "Attachment Parenting" proposed by William Sears, based on the "Theory of Attachment" developed by John Bowlby and claimed by various contemporary authors who defend the practice of co-sleeping, I will analyze the psychoanalytic concepts against it through an exploratory bibliographical revision that seeks to deepen the knowledge about co-sleeping.

Post-transplant monitoring of NK cell counts as a simple approach to predict the occurrence of opportunistic infection in liver transplant recipients.

BACKGROUND: Monitoring of peripheral blood lymphocyte subpopulation (PBLS) counts might be useful for estimating the risk of infection after liver transplantation (LT). METHODS: We prospectively measured total lymphocyte and PBLS counts at baseline and post-transplant months 1 and 6 in 92 LT recipients. PBLS were enumerated by single-platform 6-color flow cytometry technology. Areas under receiver operating characteristic (ROC) curves were used to evaluate the accuracy of different PBLS for predicting cytomegalovirus (CMV) disease and overall opportunistic infection (OI). Adjusted hazard ratios (aHRs) for both outcomes were estimated by Cox regression. RESULTS: After a median follow-up of 730.0 days, 29 patients (31.5%) developed 38 episodes of OI (including 22 episodes of CMV disease). The counts of CD3(+) , CD4(+) , and CD8(+) T cells, and CD56(+) CD16(+) natural killer (NK) cells at month 1 were significantly lower in patients subsequently developing OI. The NK cell count was the best predictive parameter (area under ROC curve for predicting CMV disease: 0.78; P-value = 0.001). Patients with an NK cell count <0.050 × 10(3) cells/µL had higher cumulative incidences of CMV disease (P-value = 0.001) and overall OI (P-value <0.001). In the multivariate models, an NK cell count <0.050 × 10(3) cells/µL at month 1 post transplantation remained as an independent risk factor for CMV disease (aHR: 5.54; P-value = 0.003) and overall OI (aHR: 7.56; P-value <0.001). CONCLUSION: Post-transplant kinetics of NK cell counts may be used as a simple and affordable proxy to the cell-mediated immunity status in LT recipients and to their associated risk of OI.

Activated Regulatory T Cells Expressing CD4(+)CD25(high)CD45RO(+)CD62L(+) Biomarkers Could Be a Risk Factor in Liver Allograft Rejection.

Activated regulatory T cells (aTregs) are nowadays a hot topic in organ transplantation to establish their role during acute rejection (AR) episodes. The aim of this multi-center study was to monitor the frequency of aTregs within the first year after transplantation in a cohort of first-time liver transplant recipients enrolled from 2010 to 2012. aTregs frequency was analyzed by means of flow cytometry. Patients who had AR showed higher levels of aTregs during first year after transplantation in comparison with patients who did not have higher levels. High levels of aTregs in liver recipients might be used as a biomarker of AR; however, further studies must be done to address the potential role of aTregs as biomarkers of AR in liver transplantation.

[Prenatal diagnosis and novel mutation in X-linked chronic granulomatous disease]. / Diagnóstico prenatal y nueva mutación en enfermedad granulomatosa crónica ligada al cromosoma X.

BACKGROUND: Chronic Granulomatous Disease (CGD) is a rare primary immunodeficiency caused by the alteration of the enzyme complex NADPH oxidase, which affects the phagocytic function. CGD patients are susceptible to recurrent infections mainly caused by bacteria and/or fungi. METHODS: We studied a 6 year-old boy with suspicion of CGD. The diagnosis was confirmed based on the functional study of NADPH oxidase. Simultaneously, the second pregnancy of the mother was reported and genetic counselling was requested. RESULTS: We identified a new disease-causing mutation by direct sequencing of the CYBB gene (X-linked CGD). The prenatal study resulted in the identification of the same mutation in the foetus. COMMENTS: Molecular genetics characterisation of CGD is needed to obtain an accurate diagnosis of the disease and to offer prenatal diagnosis and genetic counselling in future pregnancies.

Esplenomegalia familiar como manifestación clínica del síndrome linfoprolifetativo autoinmune / Familial splenomegaly as a first clinical sign of autoimmune lymphoproliferative syndrome

Introducción: El síndrome linfoproliferativo autoinmune es la consecuencia de un defecto genético que compromete la apoptosis de los linfocitos. La linfoproliferación se manifiesta por adenomegalias y/o esplenomegalia crónica. El diagnóstico requiere demostrar el defecto de la apoptosis linfocitaria y el aumento de linfocitos T dobles negativos (TDN) que carecen de CD4 y CD8. Existe riesgo de desarrollar linfomas y enfermedades autoinmunes, sobre todo citopenias. Métodos: Estudiamos a un niño de 14 años con esplenomegalia de varios años de evolución con antecedentes familiares de esplenomegalia y adenomegalias. Se analiza el fenotipo de los linfocitos T y el defecto molecular del gen TNFRSF6 en el niño, su hermana y el padre. Se estudia el defecto de la apoptosis en el niño y su padre. Resultados: En el niño y su padre se confirman el defecto de la apoptosis de los linfocitos, el aumento de linfocitos dobles negativos (el 18 y 5%, respectivamente) y la misma mutación del gen TNFRSF6. La hermana presenta la misma mutación con un 16% de linfocitos doblemente negativos. Comentarios: La esplenomegalia crónica familiar puede ser la única manifestación del síndrome linfoproliferativo autoinmune (AU)

Background: The autoimmune lymphoproliferative syndrome (ALPS) is caused by genetic defect in lymphocyte apoptosis. Chronic lymphadenopathy and splenomegaly are the consequence of lymphoproliferation. The diagnosis is based on the assessment of the defective lymphocyte apoptosis and the identification of lymphocyte T subset that are double negative (CD4-CD8-). The susceptibility to lymphoma and autoimmune diseases, mainly blood cytopenias is increased. Methods: We studied a 14 year-old boy with chronic splenomegaly and familial history of splenomegaly and lymphadenopathy. T lymphocyte phenotypes, and molecular defect of TNFRSF6 gene were studied in the child, his sister and his father. Lymphocyte apoptosis was also analysed in the child and his father. Results: The boy and his father showed in vitro apoptosis defects, an increased number of double negative T lymphocytes (18% and 5%, respectively) and the same mutation in the TNFRSF6 gene. His sister had 16% of double negative T lymphocytes and the mutation in the TNFRSF6 gene. Comments: Chronic familial splenomegaly can be the only clinical sign of autoimmune lymphoproliferative syndrome (AU)

[Familial splenomegaly as a first clinical sign of autoimmune lymphoproliferative syndrome]. / Esplenomegalia familiar como manifestación clínica del síndrome linfoprolifetativo autoinmune.

BACKGROUND: The autoimmune lymphoproliferative syndrome (ALPS) is caused by genetic defect in lymphocyte apoptosis. Chronic lymphadenopathy and splenomegaly are the consequence of lymphoproliferation. The diagnosis is based on the assessment of the defective lymphocyte apoptosis and the identification of lymphocyte T subset that are double negative (CD4-CD8-). The susceptibility to lymphoma and autoimmune diseases, mainly blood cytopenias is increased. METHODS: We studied a 14 year-old boy with chronic splenomegaly and familial history of splenomegaly and lymphadenopathy. T lymphocyte phenotypes, and molecular defect of TNFRSF6 gene were studied in the child, his sister and his father. Lymphocyte apoptosis was also analysed in the child and his father. RESULTS: The boy and his father showed in vitro apoptosis defects, an increased number of double negative T lymphocytes (18% and 5%, respectively) and the same mutation in the TNFRSF6 gene. His sister had 16% of double negative T lymphocytes and the mutation in the TNFRSF6 gene. COMMENTS: Chronic familial splenomegaly can be the only clinical sign of autoimmune lymphoproliferative syndrome.

Longitudinal analysis of immune function in the first 3 years of life in thymectomized neonates during cardiac surgery.

The purpose of this study is to evaluate the effects of neonatal thymectomy in the functional capacity of the immune system. We selected a group of 23 subjects, who had undergone thymectomy in their first 30 days of life, during an intervention for congenital heart disease. Several parameters of the immune system were evaluated during their first 3 years of life. Lymphocyte populations and subpopulations (including naive, memory and effector subpopulations), T cell receptor (TCR) Vbeta repertoire, response of T cells following in vitro stimulation by mitogen, quantification of immunoglobulins, TCR excision circles (TRECS) and interleukin (IL)-7 were measured. We found that neonatal thymectomy produces long-term diminution in total lymphocyte counts, especially in naive CD4+ and CD8+ T cells. Additionally, TRECS were decreased, and plasma IL-7 levels increased. A statistically significant negative correlation was found between absolute CD4+ T cells and IL-7 (r = -0.470, P = 0.02). The patients did not suffer more infectious events than healthy control children, but thymectomy in neonates resulted in a significant decrease in T lymphocyte levels and TRECS, consistent with cessation of thymopoiesis. This could produce a compromise in immune function later in life, especially if the patients suffer T cell depletion and need a reconstitution of immune function.

Pharmacoepidemiological approach to the predisposing factors for highly active antiretroviral therapy failure in an HIV-positive cohort from Cordoba City (Argentina) 1995-2005.

Highly active antiretroviral therapy (HAART) restores immunity, avoids resistance and delays disease progression. Nonetheless, adverse medicament reactions (AMRs) and therapeutic failure (TF) are still deleterious events. Consequently, their predisposing factors should be evaluated. Data from 181 men and 28 women of an Argentinean cohort (1995-2005) were collected and analysed by logistic regression, studying 63 schemes (15 active principles). The AMRs were the main cause of scheme change, followed by TF and medicament simplification, without influence of age and sex. Twenty-nine schemes exhibited TF at least once. Compared with zidovudine-lamivudine-nevirapine (success: >75%), the following schemes fail more frequently (P < 0.01): pre-HAART (8-fold), indinavir-containing ones (30-fold) and retrotranscriptase inhibitors with > or =3 protease inhibitors (11-fold). Inadequate patient adherence preceded failure (>95%), but not successful treatments, with a strong AMR-TF association (P < 0.005). Although some schemes had inherently increased TF, low adherence, drug toxicity and TF were critically interrelated, interfering with HAART goals.

Effects of the herbicide Roundup on freshwater microbial communities: a mesocosm study.

The impact of the widely used herbicide glyphosate has been mainly studied in terrestrial weed control, laboratory bioassays, and field studies focusing on invertebrates, amphibians, and fishes. Despite the importance of phytoplankton and periphyton communities at the base of the aquatic food webs, fewer studies have investigated the effects of glyphosate on freshwater microbial assemblages. We assessed the effect of the commercial formulation Roundup using artificial earthen mesocosms. The herbicide was added at three doses: a control (without Roundup) and two treatments of 6 and 12 mg/L of the active ingredient (glyphosate). Estimates of the dissipation rate (k) were similar in the two treatments (half-lives of 5.77 and 7.37 d, respectively). The only two physicochemical parameters showing statistically significant differences between treatments and controls were the downward vertical spectral attenuation coefficient kd(lambda), where lambda is wavelength, and total phosphorus concentration (TP). At the end of the experiment, the treated mesocosms showed a significant increase in the ratio kd(490 nm)/k(d)(550 nm) and an eightfold increase in TP. Roundup affected the structure of phytoplankton and periphyton assemblages. Total micro- and nano-phytoplankton decreased in abundance in treated mesocosms. In contrast, the abundance of picocyanobacteria increased by a factor of about 40. Primary production also increased in treated mesocosms (roughly by a factor of two). Similar patterns were observed in the periphytic assemblages, which showed an increased proportion of dead: live individuals and increased abundances of cyanobacteria (about 4.5-fold). Interestingly, the observed changes in the microbial assemblages were captured by the analysis of the pigment composition of the phytoplankton, the phytoplankton absorption spectra, and the analysis of the optical properties of the water. The observed changes in the structure of the microbial assemblages are more consistent with a direct toxicological effect of glyphosate rather than an indirect effect mediated by phosphorus enrichment.

[Strongyloides stercoralis and HIV: a case report of an indigenous disseminated infection from non-endemic area]. / Strongyloides stercoralis y VIH: un caso de infección diseminada en una zona no endémica.

Strongyloides stercoralis affects people who live in the tropics, but it also extends to temperate regions where its low incidence rate may lead to misdiagnose it. Inside the host this nematode may remain silent for many years, nevertheless, in patients infected with HIV its reactivation leads to a disseminated infection in ectopic sites. This report presents the case of a 34 years-old man infected with HIV who lived in a highland area in the province of Córdoba, Argentina. He was admitted to the hospital because of a complicated Pneumocystis jiroveci pneumonia, receiving trimethoprim-sulfamethoxazole and prednisone treatment. A few days after admission his conditions deteriorated badly, a sputum examination revealed filariform larvae of S. stercoralis, he was then given oral ivermectin medication (200 microg/kg/d). Malabsorption and ileus were installed and he died from multiorganic failure. The evolution of HIV infection to AIDS, a steroids treatment and therapy failure despite oral ivermectin triggered larvae proliferation and lead to disseminated hyperinfection until he died. This report presents not only the first case of disseminated strongyloidiasis detected in our Hospital but also an indigenous infection acquired in a non-endemic area. An awareness of an increased predisposition to this infection, especially in immunocompromised patients with malabsorption and ileus is of paramount importance, since failure to initiate appropriate therapy can lead to catastrophic outcomes, as illustrated in this case report.

Toward gene therapy for human CD3 deficiencies.

The CD3 subunits of the T cell receptor-CD3 complex (TCR-CD3) help to regulate surface TCR-CD3 expression, and participate in signal transduction leading to intrathymic selection and peripheral antigen recognition by T lymphocytes. Humans who lack individual CD3 chains show impairments in the expression and activation-induced downregulation of TCR-CD3, and the defective immune responses that result may be lethal. We have investigated delivery of a normal CD3 chain to treat disorders of this type. Retroviral transduction of CD3gamma into CD3gamma-deficient peripheral blood T lymphocytes from two unrelated patients selectively corrected the observed TCR-CD3 expression and downregulation defects, but unexpectedly seemed to cause adverse effects that can be explained by an autoreactive recognition mechanism. These data support the feasibility of gene therapy for human CD3 deficiencies, but also suggest that gene transfer into postthymic lymphocytes carrying mutations on T cell recognition or activation pathways may disrupt their intrathymic calibration and become harmful to the host.

Phylogeography of crossbills, bullfinches, grosbeaks, and rosefinches.

Mitochondrial cytochrome b (cyt b) from 24 Carduelini species including crossbills, bullfinches, grosbeaks, rosefinches, and other related, but not conclusively classified species, was sequenced. These sequences were also compared with all the available sequences from the genera Carduelis, Serinus, and Passer. Phylogenetic analyses consistently gave the same groups of finches and the calculated divergence times suggest that speciation of the studied species occurred between 14 and 3 million years ago (Miocene-Pliocene), appearing before the Passer, Carduelis, and Serinus genera. Pleistocene glaciations may have been important in sub-speciation. Crossbills are integrated within the genus Carduelis, and within redpolls; the common crossbill shows subspeciation with Loxia japonica in the Pleistocene epoch. Pinicola enucleator groups together with bullfinches and is probably the ancestor of the group. Hawfinch is only distantly related to the studied groups, and might either represent an isolated genus or be related to the New World genus Hesperiphona. The grosbeak genera Eophona and Mycerobas are clearly sister groups, and species belonging to the former might have given rise to Mycerobas species. The isolated (in classification) Uragus sibiricus and Haematospiza sipahi are included within the genus Carpodacus (rosefinches); Carpodacus nipalensis is outside the genus Carpodacus in the molecular analyses and might be an isolated species or related to the genus Montifringilla.

The origin of Palestinians and their genetic relatedness with other Mediterranean populations.

The genetic profile of Palestinians has, for the first time, been studied by using human leukocyte antigen (HLA) gene variability and haplotypes. The comparison with other Mediterranean populations by using neighbor-joining dendrograms and correspondence analyses reveal that Palestinians are genetically very close to Jews and other Middle East populations, including Turks (Anatolians), Lebanese, Egyptians, Armenians, and Iranians. Archaeologic and genetic data support that both Jews and Palestinians came from the ancient Canaanites, who extensively mixed with Egyptians, Mesopotamian, and Anatolian peoples in ancient times. Thus, Palestinian-Jewish rivalry is based in cultural and religious, but not in genetic, differences. The relatively close relatedness of both Jews and Palestinians to western Mediterranean populations reflects the continuous circum-Mediterranean cultural and gene flow that have occurred in prehistoric and historic times. This flow overtly contradicts the demic diffusion model of western Mediterranean populations substitution by agriculturalists coming from the Middle East in the Mesolithic-Neolithic transition.

The Old World sparrows (genus Passer) phylogeography and their relative abundance of nuclear mtDNA pseudogenes.

The phylogenetic relationships of genus Passer (Old World sparrows) have been studied with species covering their complete world living range. Mitochondrial (mt) cyt b genes and pseudogenes have been analyzed, the latter being strikingly abundant in genus Passer compared with other studied songbirds. The significance of these Passer pseudogenes is presently unclear. The mechanisms by which mt cyt b genes become pseudogenes after nuclear translocation are discussed together with their mode of evolution, i.e., transition/transversion mitochondrial ratio is decreased in the nucleus, as is the constraint for variability at the three codon positions. However, the skewed base composition according to codon position (in 1st position the percentage is very similar for the four bases, in 2nd position there are fewer percentage of A and G and more percentage of T, and in 3rd codon position fewer percentage of G and T and is very rich in A and C) is maintained in the translocated nuclear pseudogenes. Different nuclear internal mechanisms and/or selective pressures must exist for explaining this nuclear/mitochondrial differential DNA base evolutive variability. Also, the phylogenetic usefulness of pseudogenes for defining relationships between closely related lineages is stressed. The analyses suggest that the primitive genus Passer species comes from Africa, the Cape sparrow being the oldest: P. hispaniolensis italiae is more likely conspecific to P. domesticus than to P. hispaniolensis. Also, Passer species are not included within weavers or Estrildinae or Emberizinae, as previously suggested. European and American Emberizinae sparrows are closely related to each other and seem to be the earliest species that radiated among the studied songbirds (all in the Miocene Epoch).

HLA molecular markers in Tuvinians: a population with both Oriental and Caucasoid characteristics.

HLA class I and class II alleles have been studied for the first time in the Turkish-speaking Tuvinian population, which lives in Russia, North of Mongolia and close to the Altai mountains. Comparisons have been done with about 11000 chromosomes from other worldwide populations, and extended haplotypes, genetic distances, neighbor joining dendrograms and correspondence analyses have been calculated. Tuvinians show an admixture of Mongoloid and Caucasoid characters, the latter probably coming from the ancient Kyrgyz background or, less feasibly, more recent Russian Caucasoid admixture. However, Siberian population traits are not found and thus Tuvinians are closer to Central Asian populations. Siberians are more related to Na-Dene and Eskimo American Indians; Amerindians (from nowadays Iberian--America) are not related to any other group, including Pacific Islanders, Siberians or other American Indians. The 'more than one wave' model for the peopling of the Americas is supported.

Role of Nijmegen breakage syndrome protein in specific T-lymphocyte activation pathways.

Nijmegen breakage syndrome (NBS) is a genetic disorder characterized by immunodeficiency, microcephaly, and "bird-like" facies. NBS shares some clinical features with ataxia telangiectasia (AT), including increased sensitivity to ionizing radiation, increased spontaneous and induced chromosome fragility, and strong predisposition to lymphoid cancers. The mutated gene that results in NBS codes for a novel double-stranded DNA break repair protein, named nibrin. In the present work, a Spanish NBS patient was extensively characterized at the immunological and the molecular DNA levels. He showed low CD3(+)-cell numbers and an abnormal low CD4(+) naive cell/CD4(+) memory cell ratio, previously described in AT patients and also described in the present report in the NBS patient. The proliferative response of peripheral blood lymphocytes in vitro to mitogens is deficient in NBS patients, but the possible link among NBS mutations and the abnormal immune response is still unknown.

HLA alleles and haplotypes in the Turkish population: relatedness to Kurds, Armenians and other Mediterraneans.

Turkish and Kurdish HLA profiles are studied for the first time. The comparative study of their allele frequencies, characteristic haplotypes, genetic distances with other Mediterraneans is complemented by neighbor-joining dendrograms and correspondence analyses. Turks, Kurds, Armenians, Iranians, Jews, Lebanese and other (Eastern and Western) Mediterranean groups seem to share a common ancestry: the older "Mediterranean" substratum. No sign of the postulated Indo-European (Aryan) invasion (1200 B.C.) is detected by our genetic analysis. It is concluded that this invasion, if occurred, had a relatively few invaders in comparison to the already settled populations, i.e. Anatolian Hittite and Hurrian groups (older than 2000 B.C.). These may have given rise to present-day Kurdish, Armenian and Turkish populations.