Appropriateness of newborn screening for classic galactosaemia: a systematic review.

Currently, there is no universal agreement on galactosaemia screening, fundamentally because of the risk-benefit uncertainties. We conducted two exhaustive systematic searches in the main electronic databases (PubMed, Embase, Cochrane, etc.) to recover relevant information about the disease and screening test/s in order to support decision making in Spain. All of the 45 studies identified that covered disease issues were retrospective case series or cross-sectional analysis (level-4 evidence). Studies consistently found that the majority of patients presented characteristic symptomatology before diagnosis. Long term disabilities were not significantly correlated with age of diagnosis, onset of dietary restriction or strict diet compliance. The five studies that provided accuracy data used different cut-off points and verification tests, and thus differed in their definitions of a positive case (level-3b evidence). The estimated sensitivity was 100 % and the specificity 99.9 %. The false-positive rate ranged from 0.0005 % to 0.25 %, and the PPV from 0 % to 64.3 %. The comparative clinical effectiveness in relation to not screening or implementation of other programs is unknown. In summary, existing evidence remains insufficient to establish the appropriateness of newborn screening for galactosaemia screening, although health benefits could be expected if early diagnosis and treatment is achieved. If screening is implemented in Spain, it would be important that a pilot programme be implemented to assess false positive rate and ensure that early diagnosis is not delayed.

Use of macroalgae stored in an Environmental Specimen Bank for application of some European Framework Directives.

Different European Framework Directives have established a series of objectives for conservation of the coast, and monitoring tools must be made available to test compliance with these aims. In the present study the use of macroalgae deposited in an Environmental Specimen Bank was evaluated as a possible environmental tool for monitoring the coastal ecosystem. The concentrations of Al, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, V and Zn in three species of the genus Fucus (Fucus spiralis, Fucus vesiculosus and Fucus ceranoides) were measured at sampling sites distributed along the coast of Galicia (NW Spain). In the period 1990-2001, the concentrations of the metals were higher in 1990 than in 2001, with the exception of: i) Mn in F. ceranoides and Fe in F. spiralis-F. vesiculosus, for which there were no differences between the sampling periods, and ii) Zn in F. vesiculosus and Fe in F. ceranoides, for which the concentrations were higher in 2001 than in 1990. In the period 2001-2007 concentrations of the metals were more stable, especially in F. ceranoides (e.g. Al, Fe, Hg, Ni and V). The concentrations were also more stable vin F. vesiculosus in 2005 (i.e. Al, Cr, Fe, Mn and Zn). The population density distributions are consistent with the results of the statistical tests. The results indicate that macroalgae of the genus Fucus may be useful for applying different European Framework Directives, given that the macroalgae are sufficiently sensitive to changes in concentrations of metals, and may be suitable for long-term monitoring and used for the detection of increased concentrations of metals (real-time monitoring).

CCR6-deficient mice have impaired leukocyte homeostasis and altered contact hypersensitivity and delayed-type hypersensitivity responses.

CCR6 expression in dendritic, T, and B cells suggests that this beta-chemokine receptor may regulate the migration and recruitment of antigen-presenting and immunocompetent cells during inflammatory and immunological responses. Here we demonstrate that CCR6-/- mice have underdeveloped Peyer's patches, in which the myeloid CD11b+ CD11c+ dendritic-cell subset is not present in the subepithelial dome. CCR6-/- mice also have increased numbers in T-cell subpopulations within the intestinal mucosa. In 2,4-dinitrofluorobenzene-induced contact hypersensitivity (CHS) studies, CCR6-/- mice developed more severe and more persistent inflammation than wild-type (WT) animals. Conversely, in a delayed-type hypersensitivity (DTH) model induced with allogeneic splenocytes, CCR6-/- mice developed no inflammatory response. The altered responses seen in the CHS and DTH assays suggest the existence of a defect in the activation and/or migration of the CD4(+) T-cell subsets that downregulate or elicit the inflammation response, respectively. These findings underscore the role of CCR6 in cutaneous and intestinal immunity and the utility of CCR6-/- mice as a model to study pathologies in these tissues. This article was published online in advance of the print edition. The date of publication is available from the JCI website, http://www.jci.org.

Expression of CCR9 beta-chemokine receptor is modulated in thymocyte differentiation and is selectively maintained in CD8(+) T cells from secondary lymphoid organs.

Chemokines appear to have an important role in the seeding of lymphoid progenitors in the thymus, the regulation of the coordinated movements of the maturing T cells within this organ, and the egress of the resulting naive T cells to secondary lymphoid organs. CCR9, the specific receptor for the beta-chemokine TECK/CCL25, is selectively expressed in thymus, lymph node, and spleen. Using a specific anti-CCR9 polyclonal antibody, K629, and a semiquantitative reverse transcriptase-polymerase chain reaction procedure, a detailed study of CCR9 expression in the thymus and secondary lymphoid organs was performed. The results show that CD4(+)CD8(+) double-positive thymocytes have the highest CCR9 expression in thymus. Single-positive CD8(+) thymocytes continue to express this receptor after abandoning the thymus as mature naive T cells, as suggested by the existence of a CD8(+)CD69(low)CD62L(high) CCR9(+) cell subset. Consistent with this, CD8(+) lymphocytes from lymph nodes, spleen, and Peyer patches express a functional CCR9, as its expression correlates with migration in response to CCL25. Conversely, CD4(+) thymocytes lose CCR9 before abandoning the thymus, and CD4(+) T cells from secondary lymphoid organs also lack CCR9 expression. Analysis of CCR9 expression in thymocytes from mice of different ages showed that CCR9 levels are affected by age, as this receptor is more abundant, and its response to CCL25 is more potent in newborn animals. Collectively, these results suggest that CCR9 has a role in thymocyte development throughout murine life, with clear differences between the CD4(+) and CD8(+) lineages.

Molecular cloning and expression of stromalin protein from Drosophila melanogaster: homologous to mammalian stromalin family of nuclear proteins.

We report the cloning of a new cDNA from Drosophila melanogaster that encodes an open reading frame of 1116 amino acid residues. It is the insect homolog of the previously reported stromalin (SA) family of nuclear proteins in mammals (Carramolino et al. [1997]. Gene 195, 151-159). Taking into account the identical domain present in all the SA family members characterized to date, we have carried out polymerase chain reaction (PCR) using degenerate oligonucleotides from the 5' and 3' ends of one of those regions of the molecule and cDNA from D. melanogaster embryos. We isolated the homologous domain of the putative Drosophila SA molecule (DSA). This cDNA fragment was used as a radiolabeled probe for screening a cDNA library from Drosophila embryos, and we have cloned a full-length cDNA for the SA homolog from an insect. The protein shows a good degree of identity with the mammalian stromalins SA-1 and SA-2, with the N and C ends being the most divergent regions of the molecule. The mRNA coding for this protein shows a molecular size of about 3.7 kb by Northern blot analysis and is essentially expressed in embryonic stage. The in situ hybridization experiments indicate that the DSA messenger is expressed mainly in neurogenic territories in the embryonic development of Drosophila. The DSA protein has been cloned and expressed in a baculovirus system, and polyclonal antibodies were generated against the recombinant molecule. Western blot analysis using these antibodies detected a main band corresponding to about 120 kDa, principally in embryos.

Rapid and high sensitivity test for direct detection of bovine herpesvirus-1 genome in clinical samples.

A procedure for direct detection of the BHV-1 genome in clinical samples, including semen, was developed. The method is based on the PCR amplification of a highly conserved DNA fragment within the glycoprotein gI sequence of the virus (323 bp between nt 1491 to nt 1814). The method is rapid and highly specific for all 27 subtypes assayed, which are included in the clinical and genetically different groups of BHV-1. The viral origin of the PCR product was assessed by Southern hybridization, with an internal probe. The method for DNA isolation from clinical samples included a fast extraction procedure with Chelex 100 resin allowing the loading of larger amounts of DNA in the PCR and in turn increasing the sensitivity of the method of detection. The level of sensitivity achieved by PCR was in the range of 1 TCID(50). This PCR assay may be an useful tool for BHV-1 monitoring in semen banks at low cost.

[Breast cancer prevention program in Valenciana community. Assessment 1992-1993]. / Programa de prevencion del cancer de mama en la Comunidad Valenciana. Resultados 1992-1993.

BACKGROUND: In 1992, a breast cancer screening program was implemented by the General Directorate of Health of the Autonomous Government of Valencia. This program was aimed to decrease the mortality caused by breast cancer in a 30% on those women submitted to the program. The program was implemented, in 1992 and 1993, and with this purpose five units of breast cancer screening were set up in five Health Areas. This paper presents our preliminary results of this program, from april 1992 to december 1993. METHODS: The program target population consisted on 125,000 healthy women aged from 45 to 65 years. Each woman recruited, a two-view (cranio-caudal and medio-lateral oblique) screen-film mammograms were performed as the primary and only film-screening examination with two years interval. Additionally and according to the criteria of the physician charged to inform the mammography a physical examination could be practised. RESULTS: 52,843 women were invited to participate. The participation rate was of 70.78%. The number of breast cancer detected was of 141, corresponding to a rate of 3.90/1000 women under screening program. CONCLUSIONS: The objectives stated, in relation to participation rate, methods applied and early time of detection, were achieved in this period of assessment.

The achaete-scute gene complex of D. melanogaster: conserved domains in a subset of genes required for neurogenesis and their homology to myc.

We have determined the nucleotide sequence of two transcription units of the AS-C and shown that their potential protein coding regions share two principal domains of homology. Both domains are conserved within the myc protein family, and one of them is highly homologous with the consensus for protein tyrosine kinase substrates. We show in addition that at least one of these domains is shared with other transcription units within the AS-C, some of which were not previously known. We suggest that the AS-C encodes several homologous polypeptides, which represent a subset of a larger gene family. We propose that each member of the family functions at an equivalent stage of a unique morphogenetic operation involving the segregation of individual neural lineages from the epidermal anlage.

Excess function hairy-wing mutations caused by gypsy and copia insertions within structural genes of the achaete-scute locus of Drosophila.

Hairy-wing (Hw) mutations cause the differentiation of extra chaetes on the cuticle of Drosophila. They are associated with modifications of the achaete-scute complex that consist, in the mutants studied, of insertions of the transposable elements gypsy (Hw1, HwBS) or copia (HwUa). gypsy and copia are inserted in achaete and scute transcribed regions, respectively. Transcription of the insertion-split genes starts at the normal site but terminates within the transposable element sequences. The RNA truncated within gypsy is 5-20 times more abundant than its homolog in wild-type flies. The abundance is reduced in Hw1 revertants and Hw1 stocks carrying su(Hw) mutations. These and other data suggest that the excess function phenotypes of Hw mutations are generated by an increase in achaete or scute transcripts.

Molecular genetics of the achaete-scute gene complex of D. melanogaster.

The achaete-scute gene complex (AS-C), involved in differentiation of the sensory chaetes of D. melanogaster, and the yellow locus have been cloned. The yellow locus is the most distal and is followed, proximally, by the achaete and the scute loci. In the scute locus (75 kb), three transcription units separated by long stretches of DNA give rise to poly(A)+ RNAs of 1.6, 1.2, and 1.6 kb. Most DNA lesions associated with scute mutations map within the presumably untranscribed DNA. Their mutant phenotypes are stronger the closer the lesions are to the structural gene of one transcript (T4 RNA). Genetic and developmental data suggest that only this RNA is fundamental for the scute function. Its transcription might be perturbed by far removed DNA lesions. A second transcript is probably implicated in the lethal of scute embryonic function, while the third transcript is unnecessary for the differentiation of most macrochaetes. Two additional polyadenylated RNAs are transcribed from the achaete (1.1 kb) and yellow (1.9 kb) loci.