Pig-to-monkey islet xenotransplantation using multi-transgenic pigs.

The generation of pigs with genetic modifications has significantly advanced the field of xenotransplantation. New genetically engineered pigs were produced on an α1,3-galactosyltransferase gene-knockout background with ubiquitous expression of human CD46, with islet beta cell-specific expression of human tissue factor pathway inhibitor and/or human CD39 and/or porcine CTLA4-lg. Isolated islets from pigs with 3, 4 or 5 genetic modifications were transplanted intraportally into streptozotocin-diabetic, immunosuppressed cynomolgus monkeys (n = 5). Immunosuppression was based on anti-CD154 mAb costimulation blockade. Monitoring included features of early islet destruction, glycemia, exogenous insulin requirement and histopathology of the islets at necropsy. Using these modified pig islets, there was evidence of reduced islet destruction in the first hours after transplantation, compared with two series of historical controls that received identical therapy but were transplanted with islets from pigs with either no or only one genetic modification. Despite encouraging effects on early islet loss, these multi-transgenic islet grafts did not demonstrate consistency in regard to long-term success, with only two of five demonstrating function beyond 5 months.

Etapas del proceso de construcción de la inteligencia sensoriomotriz en bebés argentinos / Construction stages of the sensorimotor intelligence

El objetivo del artículo es presentar los resultados correspondientes a una investigación que se enmarca en un proyecto UBACyT 2008-2010, cuyo título es ôEvaluación Nacional de la Inteligencia Sensoriomotriz a bebés de 6 a 30 mesesö. El objetivo principal de dicha investigación es conocer las etapas del proceso de construcción de la inteligencia práctica en bebés argentinos en las distintas provincias de la Argentina y la elaboración de nuevos baremos a nivel nacional para la Escala Argentina de Inteligencia Sensoriomotriz (EAIS). La muestra se encuentra compuesta por 800 niños de 6 a 30 meses de edad de las provincias de Buenos Aires y CABA, Córdoba, Entre Ríos, Santa Fe, Salta, Chaco, Misiones, Mendoza, Santa Cruz y Río Negro. No se observaron diferencias significativas entre las provincias argentinas en los niveles de desarrollo cognitivo en los bebés. No ha sido necesaria la elaboración de tablas diferenciales de baremos por región. Se presentan las tablas de baremos para la EAIS para la evaluación del desarrollo cognitivo en bebés de 6 a 30 meses de edad a nivel nacional.(AU)

Etapas del proceso de construcción de la inteligencia sensoriomotriz en bebés argentinos / Construction stages of the sensorimotor intelligence

El objetivo del artículo es presentar los resultados correspondientes a una investigación que se enmarca en un proyecto UBACyT 2008-2010, cuyo título es “Evaluación Nacional de la Inteligencia Sensoriomotriz a bebés de 6 a 30 meses”. El objetivo principal de dicha investigación es conocer las etapas del proceso de construcción de la inteligencia práctica en bebés argentinos en las distintas provincias de la Argentina y la elaboración de nuevos baremos a nivel nacional para la Escala Argentina de Inteligencia Sensoriomotriz (EAIS). La muestra se encuentra compuesta por 800 niños de 6 a 30 meses de edad de las provincias de Buenos Aires y CABA, Córdoba, Entre Ríos, Santa Fe, Salta, Chaco, Misiones, Mendoza, Santa Cruz y Río Negro. No se observaron diferencias significativas entre las provincias argentinas en los niveles de desarrollo cognitivo en los bebés. No ha sido necesaria la elaboración de tablas diferenciales de baremos por región. Se presentan las tablas de baremos para la EAIS para la evaluación del desarrollo cognitivo en bebés de 6 a 30 meses de edad a nivel nacional.

Resultados preliminares de 7 provincias argentinas en el marco de una propuesta de Evaluación Nacional de la Inteligencia Sensoriomotriz a bebes de 6 a 30 meses / Preliminary results from 7 provinces of argentina within a proposition of a National Assessment of Sensory Motor Intelligence for argentinean babies of 6 to 30 months old

El objetivo es presentar los resultados preliminares de siete provincias argentinas correspondientes a una investigación que se enmarca en un proyecto UBACyT 2008-2010, cuyo título es "Evaluación Nacional de la Inteligencia Sensoriomotriz a bebés de 6 a 30 meses". El objetivo principal de dicha investigación es conocer las etapas del proceso de construcción de la inteligencia práctica en bebés argentinos en las distintas provincias de la Argentina y la elaboración de baremos a nivel nacional y por cada provincia, administrando la Escala Argentina de Inteligencia Sensoriomotriz (EAIS). La muestra se encuentra compuesta por 109 niños de las provincias de Santa Fe, Santa Cruz, Río Negro, Entre Ríos, Salta, Córdoba y Buenos Aires. En el análisis de los datos, se observó que el 67% de los niños evaluados obtuvieron resultados normales, 16.5% de riesgo y 3.7% de retraso. Es decir, el 20% de los niños evaluados obtuvieron resultados de riesgo y retraso

The aim of this article is to present the preliminary results from seven provinces of Argentina within an investigation belonging to UBACyT 2008-2010, which title is "National assessment of the sensorymotor intelligence in babies of 6 to 30 months old". The main objective of the research is to assess the periods involved in the construction process of the practical intelligence in Argentinean babies from different provinces, as well as the elaboration of national and provincial percentiles, administrating the Argentinean scale of Sensorymotor intelligence (EAIS). The sample is composed by 109 babies from the Argentinean provinces of Santa Fe, Salta Cruz, Río Negro, Entre Ríos, Salta, Córdoba y Buenos Aires. In the analysis of the data, it was observed that 67% of the babies obtained normal results, 16.5% obtained results indicating risk and 3.7% delayed. In other words, 20% of the assessed babies obtained results indicating risk and delayed in the sensorymotor intelligence

Investigation of lymphocyte depletion and repopulation using alemtuzumab (Campath-1H) in cynomolgus monkeys.

As the target CD52 molecule is expressed on erythrocytes of most nonhuman primate strains, using alemtuzumab in these species would cause massive hemolysis. Six cynomolgus monkeys of Indonesian origin, screened by agglutination assay for absence of CD52 on erythrocytes, were administered alemtuzumab in a cumulative dose to a maximum of 60 mg/kg. In two monkeys, mycophenolate mofetil (MMF) was added as maintenance therapy. Complete depletion of T and B lymphocytes (>99.5%) was achieved with 20 mg/kg alemtuzumab and was more profound than in monkeys treated with antithymocyte globulin (n = 5), as quantified by flow cytometry. Repopulation was suppressed by weekly injections of 10 mg/kg. Without MMF, repopulation of CD20(+)B cells and CD8(+)T cells was complete within 2 and 3 months, respectively, and repopulation of CD4(+)T cells was 67% after 1 year. MMF significantly delayed CD4(+)T-cell repopulation. Among repopulating CD4(+) and CD8(+) T cells, a phenotypic shift was observed from CD45RA(hi)CD62L(hi) naïve cells toward CD45RA(lo)CD62L(lo) effector memory cells. In lymph nodes, the depletion of naïve cells was more profound than of memory cells, which may have initiated a proliferation of memory cells. This model offers opportunities to investigate lymphocyte depletion/repopulation phenomena, as well as the efficacy of alemtuzumab in preclinical transplantation models.

Epilepsia por displasia cortical refractaria al tratamiento farmacológico: a propósito de un caso / Epilepsy caused by cortical dysplasia intractable with drug treatment: apropos of a case

La epilepsia refractaria al tratamiento farmacológico es un tema en revisión debido al surgimiento de nuevos fármacos antiepilépticos, de técnicas de neuroimagen y a la posibilidad de cirugía de la epilepsia Con el objetivo de enfatizar la importancia de la búsqueda etiológica para adecuar el tratamiento y modificar el pronóstico de estos niños, se presenta un caso clínico en una niña de 8 años, portadora de epilepsia refractaria desde los primeros años de vida. El uso de RM permitió establecer que la misma era originada en una zona de displasia cortical. Se realizó cirugía de la epilepsia estableciéndose un pronóstico favorable a mediano plazo(AU)

[Occupational stress in health care personnel]. / Estrés ocupacional en personal de salud.

BACKGROUND: Occupational stress results from the interaction of multiple risk factors, such as the physical environment, biological function disturbances, work content and organization and diverse psychosocial components. AIM: To study the frequency of symptoms and the main sources of job stress, perceived by professional and non professional health care workers and to compare gender differences. SUBJECTS AND METHODS: A group of specially designed, self-administered, questionnaires adapted by the main author and independently validated, were applied. The results of two symptoms, one work satisfaction, and one job stress scales are reported. RESULTS: One hundred sixteen women and 89 men were studied, 143 are professionals and 62 non professionals (clerical and nurse auxiliaries). Forty percent had symptoms of job stress and 82 subjects were defined as "probable cases", according to the ratings on the symptom scales. There were significant differences in the frequency of symptoms between professionals and non professionals (34.3 and 69.2% respectively). The main differences in symptoms, sources of job stress and dissatisfaction were more closely associated to the occupational level than to gender. CONCLUSIONS: A high frequency of job stress symptoms was observed in this sample of health care workers and the risk of occupational stress varies more with the occupational level than with gender.

[Recent alcohol and drug consumption in victims of work related accidents]. / Consumo reciente de alcohol y drogas en accidentes del trabajo.

BACKGROUND: Twenty to twenty two percent of work related accidents have a direct or indirect relationship with recent alcohol use. AIM: To detect recent alcohol or drug use in patients who required hospitalization for severe work related accidents. PATIENTS AND METHODS: Blood and urine samples were obtained from patients admitted to a hospital within 6 hours of an accident severe enough to require hospitalization. Blood alcohol and urinary metabolites of cocaine, marijuana, amphetamines and benzodiazepines were measured. Patient identity was kept confidential throughout the study. Once patients were stabilized, they were interviewed by a psychologist that was blind to laboratory results. RESULTS: Two hundred thirty men and 18 women were studied and 29% (30% of men and 17% of women) had recent use of alcohol or drugs. The most common substances were alcohol in 15% and benzodiazepines in 13%. Use of alcohol or drugs was found in 21% of professionals, in 33% of other occupational levels, half of agricultural workers and 25% of commerce workers. Only two of 60 positive cases, admitted substance use during the interview. Most of negative cases admitted the use of alcohol or other drugs during the month prior to the accident. CONCLUSIONS: Recent use of alcohol and drugs is frequent among severe work related accident victims. Probably, a high proportion of these subjects are problem drinkers. The combination of alcohol and benzodiazepines is an additional risk factor.

Molecular epidemiology of autoimmune thyroid disease.

This paper presents preliminary data regarding the prevalence and risk factors for autoimmune thyroid disease in IDDM probands ascertained from the Children's Hospital of Pittsburgh IDDM Registry for 1950-1965 (n = 669). Living IDDM probands who participated in the 1990 follow-up survey (n = 380) were recruited for the Familial Autoimmune and Diabetes Study. Siblings and parents were also invited to participate. To date, 255 IDDM probands and 597 parents and siblings have been evaluated. The diagnosis of autoimmune thyroid disease was based on a clinical evaluation, medical history, and laboratory determinations. Graves disease was rare in this cohort (n = 5). However, Hashimoto's thyroiditis was common among women. Prevalence rates ranged from 54% for IDDM women age < 40 years to 75% for those > 50 years. Corresponding age-specific estimates for female relatives were 22% and 44%, respectively. Approximately one-half of the Hashimoto's individuals were euthyroid; they were more likely to have other autoantibodies and a positive family history than those who were hypothyroid or had no thyroid disease. Genetic analyses revealed a 2-fold increase in DQA1*0501-DQB1*0201 among the Hashimoto's compared to the non-Hashimoto's haplotypes. These findings suggested that Hashimoto's thyroiditis was common in IDDM families, which may be due, in part, to common disease susceptibility genes.

Emotional and psychosocial factors in burn patients during hospitalization.

This is a preliminary study of psychological symptoms, prior psychosocial adjustment and severity of injury in 25 consecutive burn patients. All had suffered burns in work-related accidents and were evaluated by the psychiatric consulation-liaison team during their initial hospitalization in the Burn Unit of the Hospital del Trabajador, Santiago. All patients were clinically evaluated and the following instruments were applied: Hamilton Scales of Anxiety (HAM-A) and Depression (HAM-D), Goldberg's General Health Questionnaire (GHQ-28), Cloninger's Tridimensional Personality Questionnaire (TPQ) and a modified version of a Schooler's Adjustment Scale. No significant correlation was found between extent and severity of burns and anxious or depressive symptoms. However, all patients had burns that varied from mild to moderate; there were no severe and extensive injuries. A significant correlation was found between anxiety and the harm avoidance dimension in the TPQ, as would be expected. Patients with poor psychosocial adjustment presented with greater anxiety symptoms. A significant negative correlation was also found between degree of anxiety symptoms and income level. The main findings are in agreement with current literature. They emphasize the relevance of prior psychosocial adjustment and personality as probable factors of psychological symptoms. The need to control pain as a variable is also pointed out.

Altered CYP21 genes in HLA-haplotypes associated with congenital adrenal hyperplasia (CAH): a family study.

Disorders of the CYP21 gene, which is located within the major histocompatibility complex on the short arm of chromosome 6, are the leading causes of congenital adrenal hyperplasia (CAH). The coding gene and a highly homologous pseudogene are tandemly arranged with the two genes for the fourth component of complement (C4A and C4B). To analyse the prevalence rates of mutations of the CYP21 genes and the segregation of the CYP21 genes with their corresponding human leucocyte antigen (HLA)-haplotypes, 21 families with one or two children with the severe form of 21-hydroxylase deficiency were studied. Mutations of the CYP21 gene on their corresponding HLA-haplotype were detected by hybridisation of polymerase chain reaction (PCR)-amplified genomic DNA with sequence-specific oligonucleotides and solid phase direct sequencing. Our study has shown the following. (1) A single basepair mutation (A-->G or C-->G) within the second intron is the most frequent mutation leading to impaired 21-hydroxylase activity. This mutation is only detected in HLA-haplotypes associated with the salt-wasting form of CAH. (2) A large deletion of part or all of the CYP21 gene is associated with the HLA-haplotype A3, BW47, C6, DR7, DR53, DQ2 but is also observed in other HLA-haplotypes and can be detected by a simple rapid PCR restriction fragment length polymorphism method. (3) Two alleles of the coding CYP21 gene differing in a leucine codon within the first exon, (formerly described as a mutation associated with 21-hydroxylase deficiency) have been found with an equal distribution in patients with 21-hydroxylase deficiency, non-disease HLA-haplotypes and the local healthy controls.

Usage of T-cell receptor V beta chain genes in fresh and cultured tumor-infiltrating lymphocytes from human melanoma.

Tumor-infiltrating lymphocytes (TIL) freshly obtained from human malignant melanomas as well as the same TIL grown in the presence of interleukin 2 (IL2) were studied for gene expression of the T-cell receptor (TCR) variable beta regions (V beta). To perform the TCR-V beta analysis, total RNA was isolated from TIL and reverse-transcribed into cDNA, which was then amplified by PCR using 22 different 5' primers specifically recognizing the sequences of 20 V beta gene families and a 3' primer annealing to the constant region of the beta chain. The TCR-alpha constant region (C alpha) gene was co-amplified as a standard for the calculation of the percentage of each TCR-V beta gene expressed. The frequency of individual V beta regions expressed on TIL was computed from the ratio of cpm V beta to cpm C alpha for each V beta region in relation to the total of all 22 ratios. With fresh TIL obtained from 8 different melanomas, oligoclonal distribution of V beta genes expressed on TIL was observed, in comparison with a broader and unrestricted distribution seen with peripheral-blood T cells of 8 normal individuals. The oligoclonal patterns of V beta-gene expression in fresh melanoma TIL were distinct in every tumor. Several of the V beta-genes usually expressed in normal PBL were not expressed in fresh TIL in melanoma TIL cultured in the presence of IL2 and IL4 and in the absence of autologous tumor (AuTu) or antigen-presenting cells for 23 to 65 days, selection of T-cell lines expressing a restricted number of V beta genes occurred. Although in 4/5 TIL cultures this selection involved the V beta 7 gene, no relationship could be established between V beta gene expression in fresh TIL and that in T-cell lines outgrowing in long-term cultures. Selection in culture of CD3+CD8+ T-cell lines with V beta-gene expression restricted to 1 or 2 V beta families did not correlate with the presence or level of AuTu cytotoxicity mediated by these T cells. The results indicate that in TIL cultures random selection of T-cell lines with reactivity not relevant to AuTu may account for poor expression or loss of AuTu cytotoxicity by most TIL cultured long-term in the presence of cytokines and in the absence of specific antigenic stimulation.

Diabetic ketoacidosis in cystic fibrosis.

OBJECTIVE: To differentiate the insulin-dependent glucose intolerance associated with cystic fibrosis from type I diabetes mellitus in patients with cystic fibrosis. DESIGN: Patient report. SETTING: Tertiary care referral center. PARTICIPANT: An 11-year-old boy with cystic fibrosis who developed diabetic ketoacidosis. MEASUREMENT/MAIN RESULT: Biochemical, immunologic, and molecular techniques were used to support the sporadic association of type I diabetes mellitus in a patient with cystic fibrosis. Cystic fibrosis was confirmed by sweat test and further supported by the demonstration of a heterozygous deletion of the F508 locus. Evidence for the diagnosis of type I diabetes mellitus was developed from the clinical presentation of diabetic ketoacidosis with hyperglycemia, ketonemia, and ketonuria. Immunologic evidence included the demonstration of anti-insulin antibodies. The demonstration of homozygous absence of aspartic acid at position 57 of the HLA DQ-beta chain placed this child at high risk of type I diabetes mellitus. CONCLUSION: The clinical presentation and the presence of immunologic and genetic markers characteristic of type I diabetes mellitus supports the concordance of cystic fibrosis and type I diabetes mellitus in this patient.

Systemic chimerism in human female recipients of male livers.

We have previously reported data from clinical and laboratory animal observations which suggest that organ tolerance after transplantation depends on a state of balanced lymphodendritic cell chimerism between the host and donor graft. We have sought further evidence to support this hypothesis by investigating HLA-mismatched liver allograft recipients. 9 of 9 female recipients of livers from male donors had chimerism in their allografts and extrahepatic tissues, according to in-situ hybridisation and molecular techniques 10 to 19 years posttransplantation. In 8 women with good graft function, evidence of the Y chromosome was found in the blood (6/8), skin (8/8), and lymph nodes (7/8). A ninth patient whose transplant failed after 12 years from recurrent chronic viral hepatitis had chimerism in her lymph nodes, skin, jejunum, and aorta at the time of retransplantation. Although cell migration is thought to take place after all types of transplantation, the large population of migratory cells in, and the extent of their seeding from, hepatic grafts may explain the privileged tolerogenicity of the liver compared with other organs.

Autoimmunity and genetics contribute to the risk of insulin-dependent diabetes mellitus in families: islet cell antibodies and HLA DQ heterodimers.

The risk for insulin-dependent diabetes mellitus (IDDM) associated with genetic susceptibility markers at the human leukocyte antigen (HLA) DQA1 and DQB1 loci was evaluated among individuals with and those without islet cell antibodies. A total of 108 antibody-positive parents and siblings of IDDM patients from the Pittsburgh registry were identified among 1,592 who were screened. HLA-DQ molecular typing was performed on 79 of these individuals and on 78 antibody-negative relatives. There were similar proportions of homozygotes for both of the diabetogenic alleles DQA1 arginine-52 (R/R) and DQB1 non-aspartate-57 (nD/nD) among the antibody-positive and antibody-negative relatives (19.0 and 15.4%, respectively). However, subsequent development of IDDM was restricted to individuals who were both antibody positive and carried the potential to make at least one diabetogenic DQ heterodimer. A dose-response effect was observed among the antibody-positive relatives, in which two of 18 capable of generating one diabetogenic heterodimer and six of 29 generating two heterodimers became insulin requiring. Nine of 15 who were homozygous for both R/R and nD/nD, coding exclusively for diabetogenic variants, became diabetic over the course of the follow-up. With a multivariate model, the relative risk for IDDM among those with islet cell antibodies who were also R/R and nD/nD was estimated to be 229.3 compared with those lacking both, after age and sex were controlled for. The data suggest that while autoimmunity, indicated by the presence of cytoplasmic islet cell antibodies may be relatively common, it progresses only in those with variant HLA-DQ molecules.

[Workers exposed to styrene: application of a neuropsychological evaluation computer system]. / Trabajadores expuestos a estireno: aplicación de un sistema de evaluación neuropsicológica por computador.

The purpose of this study was to adapt and apply a battery of neuropsychological tests administered by computer. The study comprised three phases: 1) An existing system (NES-2) was revised and adapted (mainly in its Spanish version, since the structure and time intervals were the same). 2) A pilot group of 18 subjects was administered the five selected tests. 3) A comparative study of 36 exposed workers with 40 unexposed controls was undertaken. It was concluded that computer-administered tests are acceptable and economic. No significant differences were found between exposed. The possible reasons for the negative findings are discussed, as well as the types of future studies that need to be carried out.

Mouse NKR-P1. A family of genes selectively coexpressed in adherent lymphokine-activated killer cells.

NK cells are a subpopulation of large granular lymphocytes. They are able to recognize and lyse a wide variety of virally infected or neoplastic target cells without previous sensitization or MHC restriction. The molecules involved in target recognition and subsequent triggering of the killing process are still undefined. Recently, a 30-kDa protein highly expressed on rat NK cells and capable of mediating transmembrane signaling was identified and the gene coding for it cloned and sequenced. To better understand the role of this protein in NK cell-mediated cytotoxicity, we cloned its mouse homologue by cross-hybridization of the rat gene to a cDNA library generated from highly purified mouse lymphokine-activated NK cells. Three messages, differing in size and sequence and encoded by different genes, are specifically cotranscribed in mouse NK cells. The protein products of this gene family express the lectin-like motif characteristic of type II transmembrane molecules. Both the rat and mouse proteins have conserved tyrosine and serine residues in their cytoplasmatic portion that are potential phosphorylation sites. They also share a sequence that could be the binding site of the P56lck tyrosine kinase. These observations are consistent with the signaling function hypothesized for these proteins.

[Benzodiazepines in the treatment of anxiety. The risks and alternatives]. / Benzodiazepinas en el tratamiento de la ansiedad. Riesgos y alternativas.

The purpose of this article is to review the risks of pharmacological treatments of anxiety, mainly those due to benzodiazepines. Many mildly anxious patients do not need pharmacotherapy and psychological support is always an important component of treatment. The risks of benzodiazepine treatment increase with the doses used and the length of treatment, especially the risk of dependence. However, people who abuse benzodiazepines are a minority. Usually they also abuse alcohol and other drugs, probably reflecting underlying personality disorders. There are several alternative pharmacological treatments for anxiety. Tricyclics and MAO inhibitors are effective in panic disorders. Buspirone is useful in generalized anxiety disorder and in patients who are prone to substance abuse. All these alternative treatments may have disadvantages that need to be considered.