The nonrandom distribution of long clusters of all possible classes of trinucleotide repeats in barley chromosomes

This paper is the first to report the long-range organization of all possible classes of trinucleotide motifs in a higher plant genome. Fluorescent in situ hybridization (FISH), employing the synthetic oligonucleotides (AAC)5, (AAG)5, (AAT)5, (AGG)5, (CAC)5, (CAT)5, (CAG)5, (ACT)5, (ACG)5 and (GCC)5, was used to characterize the nonrandom and motif-dependent distribution of tandem arrays of trinucleotide repeats in the metaphase chromosomes and interphase nuclei of barley (Hordeum vulgare L.). This provided detailed information on the sequence content of barley chromatin and allowed the saturation of the physical map of all barley chromosomes. The following conclusions were also drawn: (1) Except for (AAT)5 and (GCC)5, the studied repetitive motifs have a characteristic pattern of distribution in terms of their in situ FISH signals. Some permit the accurate identification of individual chromosomes. (2) (CAG)5, (CAT)5 and (ACT)5 are not found in all barley chromosomes. (3) With the exception of (ACT)5, the remaining trinucleotide repeats occur predominantly in the heterochromatin and are largely absent from the euchromatic regions. Moreover, (CAC)5, (ACG)5 and (CAG)5 are exclusively concentrated in the centromeres. The employment of simple synthetic probes for the identification of chromosomes and genomic characterization, and their importance in studies on genome organization, function and evolution, are discussed.     

Anti-LFA-1 (CD11a) monoclonal antibody interferes with neonatal induction of tolerance to alloantigens

The injection of (C57BL/6 × BALB/c)F1 spleen cells into newborn BALB/c mice results in the induction of a specific cytotoxic T lymphocyte (CTL) tolerance to the alloantigens. On the contrary, alloreactive CD4⁺ T cells persist in the host and are still able to activate autoreactive F1 B cells to produce autoantibodies. This state of “split tolerance” is closely associated with the development of a lupus-like autoimmune syndrome. The LFA-1 integrin plays a relevant role in homing, intercellular adhesion and tranduction of co-stimulatory signals in leukocytes. Because of the beneficial effects of anti-LFA-1 monoclonal antibodies (mAb) treatment in various models of organ transplantation and autoimmune disease, we have investigated if such a treatment could interfere with the induction of neonatal tolerance or the development of the autoimmune syndrome in F1 cell-injected newborn mice. For this purpose, BALB/c mice neonatally injected with F1 cells were treated from day 1 up to day 15 with a non-cytotoxic anti-LFA-1 (CD11a) mAb. Anti-LFA-1 mAb treatment interfered with the persistence of a stable chimerism and with the establishment of CTL tolerance, as shown by rejection of allogeneic skin grafts and F1 B cells, and by a normal in vitro CTL activity against the corresponding alloantigens. As a consequence, these mice did not develop the characteristic autoimmune features seen in close association with an effective induction of CTL tolerance to alloantigens. These results stress the importance of the interactions between LFA-1 and its ligands during the neonatal induction of tolerance to alloantigens.     

Is mortalin a candidate gene for T1DM ?

Mortalin has been found to be up-regulated by 2D-protein gel analysis in isolated rodent islets exposed to cytokines. In islets from two rat strains with different sensitivity to the toxic effects of cytokines we observed a significant difference in IL-1beta mediated mortalin expression. Constitutive over-expression of rat mortalin in NIH3T3 cells reduced cellular survival in accordance with mortalin being associated to cellular senescence. Hence we consider the gene encoding for mortalin at chromosome 5q31.1 a putative candidate gene in cytokine induced beta-cell destruction. We scanned the human mortalin gene for polymorphisms and identified three novel polymorphisms. Neither the SNPs individually nor as constructed haplotypes showed disease association tested by (E)TDT in a Danish type 1 diabetes (T1DM) population. Furthermore, we tested the D5S500 microsatelite located close to 5q31.1 without finding linkage to (T1DM). In conclusion, the functional data identifying a difference in mortalin expression in IL-1beta stimulated islets between two rat strains and over-expression of mortalin in NIH3T3 cells associated with decreased viability suggests a functional role for mortalin in cytokine mediated beta cell destruction; however, the identified polymorphisms did not reveal any association in the presence of linkage disequilibrium of mortalin to T1DM in the Danish population.     

Ultrastructure of nonspecific cytotoxic cells in teleosts. I. Effector-target cell binding in a marine and a freshwater species (Seabream: Sparus aurata L., and Carp: Cyprinus carpio L.)

Background: Fish cytotoxic effectors form a cell population whose ultrastructure and properties of conjugation with target cells have not been completely established. We report the ultrastructure of the non-specific cytotoxic cells in a seawater teleost (Sparus aurata L.) and compare it to a freshwater species (Cyprinus carpio L.). Methods: Blood leucocytes were incubated with HeLa or B16 melanoma cells. Samples were processed for transmission electron microscopic study. Results: Conjugates consisting of leucocytes binding targets were regularly observed after 30 min, 1 hr, or 2 hr of incubation. In both species leucocytes binding to targets showed ultrastructural features of either monocyte-like or lymphocyte-like cells. Monocyte-like cells usually appeared flattened against the targets and seemed to enclose fragments of the target to form cytoplasmic vesicles and the content of their scarce cytoplasmic granules seemed to be delivered into these vesicles. In the seabream lymphocyte-like cells, dense cytoplasmic granules occurred only occasionally, and neither microvilli nor cell processes were present at the contact areas with the targets. In the carp, the contacts were more numerous and formed regularly interdigitating contact areas and the lymphocytes showed granules with characteristic dense and fibrillar contents. Conclusions: We conclude that seabream and carp have a leucocyte cell population with ultrastructural features of either monocytes or lymphocytes showing nonspecific cytotoxic ability. © 1994 Wiley-Liss, Inc.     

Increased Interleukin-7 Plasma Levels Are Associated with Recovery of CD4+ T Cells in HIV-Infected Children

To asses the role of interleukin 7 (IL-7) in the thymic reconstitution of CD4 T cells observed in children after successful antiretroviral therapy, a longitudinal study in five vertically HIV-1-infected children was carried out. Thymic function, IL-7 plasma levels, viral load, and T-lymphocytes subsets were determined every 2 or 3 months for about 90 months. In all the children, the drop in CD4+ T cells below 5–10% was associated with a marked increase in IL-7 plasma levels. The drastic decrease in viral load after treatment, led in all the cases to a recover of CD4 to levels higher than 30%, which was associated to an increase in thymic production of T cells and followed by a decrease in IL-7 to the normal levels. We conclude that the drop in CD4 in HIV children would induce an increase of IL-7 as part of a homeostatic mechanism. IL-7 would induce the thymus to produce new T cells to recover the normal levels of CD4 when the viral load was low and so the thymic function was not inhibited. The increase in the thymic production of new T cells recovers the CD4 population, and leads to a normalization of IL-7 levels.     

Loss of CLTRN function produces a neuropsychiatric disorder and a biochemical phenotype that mimics Hartnup disease

Hartnup disease is an autosomal recessive condition characterized by neutral aminoaciduria and behavioral problems. It is caused by a loss of B⁰AT1, a neutral amino acid transporter in the kidney and intestine. CLTRN encodes the protein collectrin that functions in the transportation and activation of B⁰AT1 in the renal apical brush bordered epithelium. Collectrin deficient mice have severe aminoaciduria. However, the phenotype associated with collectrin deficiency in humans has not been reported. Here we report two patients, an 11‐year‐old male who is hemizygous for a small, interstitial deletion on Xp22.2 that encompasses CLTRN and a 22‐year‐old male with a deletion spanning exons 1 to 3 of CLTRN. Both of them present with neuropsychiatric phenotypes including autistic features, anxiety, depression, compulsions, and motor tics, as well as neutral aminoaciduria leading to a clinical diagnosis of Hartnup disease and treatment with niacin supplementation. Plasma amino acids were normal in both patients. One patient had low 5‐hydroxyindoleacetic acid levels, a serotoninergic metabolite. We explored the expression of collectrin in the murine brain and found it to be particularly abundant in the hippocampus, brainstem, and cerebellum. We propose that collectrin deficiency in humans can be associated with aminoaciduria and a clinical picture similar to that seen in Hartnup disease. Further studies are needed to explore the role of collectrin deficiency in the neurological phenotypes.     

Characterizing the immune system after long-term undetectable viral load in HIV-1-infected children

Thirty two HIV-infected children, on highly active antiretroviral therapy (HAART) and >500 CD4⁺ T cells/mm³, were rated according to the time-course of viral load (VL) during the whole follow-up period (>18 months) in a longitudinal retrospective study. (a) uVL group: 15 children with VL below 400 copies/mL; (b) dVL group: 17 children with higher VL. The uVL group showed higher memory (CD4⁺CD45RO⁺) T cells than did dVL group, and higher number of memory activated CD4⁺CD45RO⁺HLA-DR⁺ than did control group (healthy age-matched uninfected children), whereas CD4⁺CD45RA^hi+CD62L⁺ was similar. However, TCR rearrangement excision circles (TRECs) were higher in uVL group than in dVL group. uVL Group showed CD8⁺CD45RO⁺ and CD8⁺CD45RO⁺CD38⁺ higher number than the control group, but lower than the dVL group. The percentage of CD8⁺CD45RA^hi+CD62L⁺, CD8⁺CD45RA⁺, CD8⁺CD62L⁺, and CD8⁺CD28⁺ was higher in uVL group than in dVL group, and lower than in control group. The uVL group showed higher number of activated (HLA-DR⁺CD38⁺, HLA-DR⁺, HLA-DR⁺CD38^–) CD4⁺ T cells and lower percentages of CD4⁺HLA-DR^–CD38⁺ than dVL group. In activated CD8⁺ T cell, the uVL group had lower CD8⁺HLA-DR⁺CD38⁺, CD8⁺HLA-DR⁺, and CD8⁺CD38⁺ than the dVL group. Preeffector (CD8⁺CD57^–CD28^– and CD8⁺CD45RA^–CD62L^–) T cells were lower in the uVL group than in dVL group. In the effector (CD8⁺CD57⁺, CD8⁺CD57⁺CD28^–, and CD8⁺CD45RA⁺CD62L^–) T cells, HIV-infected-children had higher values than control group. HIV-infected-children who respond to HAART had TRECs reconstitution, decreased immune activation, and lower effector CD8⁺ T cells. Moreover, successful HAART allow the increment of activated CD4⁺ T cells.     

Characterisation of the first CTX-M-10-producing isolate of Salmonella enterica serotype Virchow

Microbiological analysis of a urine sample from an outpatient with symptoms of urinary infection detected >10(5) CFU/mL urine of Salmonella enterica serotype Virchow with resistance to cefotaxime. Molecular analysis demonstrated the presence of the gene encoding CTX-M-10 beta-lactamase in this clinical isolate. This is the first report of this enzyme in Salmonella spp.     

Multiple hereditary infundibulocystic basal cell carcinomas: a genodermatosis different from nevoid basal cell carcinoma syndrome.

BACKGROUND: Infundibulocystic basal cell carcinoma is a recently described distinctive clinicopathologic variant of basal cell carcinoma. Histopathologic differential diagnosis among infundibulocystic basal cell carcinoma, trichoepithelioma, and basaloid follicular hamartoma has generated controversy in the literature. OBSERVATIONS: Members of 2 families with multiple infundibulocystic basal cell carcinomas are described. Each patient showed multiple papular lesions, mostly located on the face. No patient showed palmar pits or jaw cysts. Forty-two cutaneous lesions from 5 patients were studied histopathologically. Thirty-nine lesions were infundibulocystic basal cell carcinomas. This clinicopathologic variant of basal cell carcinoma consists of a relatively well-circumscribed basaloid neoplasm composed of buds and cords of neoplastic cells arranged in anastomosing fashion and with scant stroma. Some of the neoplastic cords contain tiny infundibular cysts filled by cornified cells with abundant melanin. Linkage analysis in family 2 was performed using polymorphic markers (D9S196, D9S280, D9S287, and D9S180), and the affected members shared the same haplotype. Loss of heterozygosity analysis was performed in 2 affected members of this family from whom tumoral DNA was available, and although these individuals were constitutively heterozygous for D9S196, they did not show loss of heterozygosity for this marker in their neoplasms. CONCLUSIONS: Multiple hereditary infundibulocystic basal cell carcinomas represent a distinctive genodermatosis different from multiple hereditary trichoepitheliomas and nevoid basal cell carcinoma syndrome. We propose clinical and histopathologic criteria to distinguish infundibulocystic basal cell carcinoma from trichoepithelioma, basaloid follicular hamartoma, and folliculocentric basaloid proliferation.     

N400 and lexical decisions: automatic or controlled processing?

OBJECTIVE: To investigate whether the N400 effect is sensitive to automatic or controlled processes. METHODS: Two experiments were performed. In one experiment, directly related word pairs were used. In the other experiment, mediated-related word pairs were used. In order to reduce controlled processes, each experiment consisted of 3 tasks: Low- and high-proportion of related pairs, and single presentation lexical decision task. RESULTS: In the first experiment, the amount of priming was equivalent for the 3 tasks. The N400 effect appeared in the high and low proportion of directly related words, but not in the single presentation task. In the second experiment, behavioral priming was also found in the 3 tasks. However, the N400 effect was observed only in the task with low proportion of related pairs. CONCLUSION: These results suggest that the N400 effect may be related to controlled processes.