Concentrations of Organochlorine Pesticides in Milk of Nicaraguan Mothers

Breast-milk samples from 101 mothers from the basin of Rio Atoya, Nicaragua, were collected on two occasions within the first trimester of lactation. Milk samples were analyzed for 13 organochlorine pesticides: (1) p,p'-dichlorophenyldichloroethylene; (2) p,p'-dichlorophenyltrichloroethane; (3) p,p'-dichlorophenyldichlorodiene; (4) α-hexachlorocyclohexane; (5) β-hexachlorocyclohexane; (6) γ-hexachlorocyclohexane; (7) δ-hexachlorocyclohexane; (8) toxaphene; (9) dieldrin; (10) endrin; (11) aldrin; (12) heptachlor; and (13) heptachlor-epoxide. Organochlorines of the dichlorodiphenylethane class (i.e., p,p'-dichlorodiphenylethane and p,p'-dichlorodiphenylethane) were found in all samples and at the highest mean concentrations observed in the study. Chemicals in the hexachlorocyclohexane family (i.e., α- and δ-hexachlorocyclohexane) were not found at all (0%), and the other hexachlorocyclohexane compounds (i.e., β > γ) were found in less than 6% of the samples. Twenty percent or less of the sample contained chlorinated cyclodienes (i.e., dieldrin > endrin > heptachlor-epoxide > heptachlor). No measurable concentrations of α-hexachlorocyclohexane, aldrin, p,p'-dichlorophenyldichlorodiene, and toxaphene were found in the breast milk samples. Analysis of variance demonstrated that only the concentration of p,p'-dichlorophenyldichloroethylene p,p'-dichlorophenyltrichloroethane, and endrin were affected significantly by maternal age. Overall, with the exception of p,p'-chlorophenyldichloroethylene, and p,p'-dichlorophenyltrichloroethane, the mean concentrations of the analyzed pesticides were low. Total p,p'-dichlorophenyltrichloroethane concentrations that exceeded the allowed daily intake set by the World Health Organisation were found in 5.9% of the samples.     

Molecularly designed alginate hydrogels susceptible to local proteolysis as three-dimensional cellular microenvironments

The development of sophisticated three-dimensional (3-D) cell culture microenvironments that recreate some of the complexity of the natural extracellular matrix (ECM) remains a challenging task. Here, the modification of alginate through partial crosslinking with a matrix metalloproteinase (MMP) cleavable peptide (proline-valine-glycine-leucine-isoleucine-glycine, PVGLIG) is described, and its use in the preparation of injectable, in situ crosslinkable hydrogel-like matrices is proposed. PVGLIG-grafted alginates were synthesized by carbodiimide chemistry and characterized. Their biological performance was evaluated by comparing the response of 3-D cultured mesenchymal stem cells (MSCs) to alginate hydrogels containing only cell-adhesion peptides (RGD-alginate) or both peptides (PVGLIG/RGD-alginate). After 1 week, cells remained essentially round within RGD-alginate, while they exhibited an elongated morphology within PVGLIG/RGD-alginate hydrogels, forming cellular networks. This suggests that cells were able to structurally reorganize the matrix, through enzymatic hydrolysis of PVGLIG residues, overcoming biophysical hydrogel resistance. As shown by gelatine-zymography, MSC presented higher activity of MMP-2 when cultured within alginate functionalized with MMP-sensitive peptide, suggesting that the cell's proteolytic phenotype was modulated by the matrix composition. Additionally, PVGLIG/RGD-alginate hydrogels were clearly degraded in cell culture. Taken together, the results demonstrate that the co-incorporation of MMP-labile peptides in cell-adhesive RGD-alginate hydrogels improved their performance as ECM analogues, providing a more dynamic and physiological 3-D cellular microenvironment.     

Pelvic exenteration and sphincter preservation in the treatment of soft tissue sarcomas.

BACKGROUND: Pelvic sarcomas are rare and there are very few effective therapeutic alternatives. A complete resection is considered the main factor associated to a good prognosis, which justifies the employment of a pelvic exenteration (PE) in selected cases. METHODS: Between 1980 and 2000, 96 PE were performed, nine of which were for sarcomas. The clinical characteristics, surgical and anatomopathological aspects and the patients' evolution were described. RESULTS: The median follow-up time was 24 months (ranging from 1 to 57 months). In relation to the sphincters preservation, at least one sphincter was preserved in five patients. There were two post-operative deaths. In the last follow-up, six patients were alive without any evidence of the disease. CONCLUSION: Pelvic exenterations should be performed for the treatment of selected cases of locally advanced pelvic sarcomas. Sphincter preservation may be performed, provided that oncological resection principles are obeyed.     

Comparison of HLA-A antigen typing by serology with two polymerase chain reaction based DNA typing methods: implications for proficiency testing

Serology has been routinely used for class I HLA typing for the selection of donors for allotransplantation. However, serology is not adequate for the assignment of all class I specificities especially when testing non-Caucasians subjects and it is necessary to adopt new strategies for routine testing. At the present time the extent of incorrect serologic HLA-A assignments in clinical testing is not known. The polymerase chain reaction (PCR) based techniques have become useful standard clinical typing methods of HLA class II alleles but most laboratories still use serology for class I typing. In this report we have compared two PCR based techniques, PCR amplification with sequencespecific primers (PCR-SSP) and PCR amplification and subsequent hybridization with sequence-specific oligonucleotide probes (PCR-SSOP), for the assignment of HLA-A specificities in 56 blood samples from patients and families serologically typed for HLA-A. This side-by-side comparison of PCR methods showed 100% correlation between them. However, serology showed 7.1% misassignments and, in an additional panel of 19 cells where serology produced equivocal results, the PCR-SSP and SSOP methods identified the correct HLA-A specificity. Our results emphasize the need to complement routine serologic testing of HLA specificities with a small number of primers designed to test HLA-A34, A36, A43, A66, A74 and A80, that are not detected with high precision by serology. We concluded that the PCR-SSP and -SSOP methods can be used in routine HLA-A typing of patients and donors for transplantation with a greater precision than serology.     

New HLA–DR haplotypes containing the DRB6 pseudogene

An unexpected probe reaction pattern was observed in two samples during HLA-DR typing by PCR-Sequence Specific Oligonucleotide Probes. In order to confirm the unusual typings, samples were analyzed by PCR-Sequence Specific Primers, cloning, and nucleotide sequencing of the second exon of the HLA-DRB-genes. The confirmed DR, DQ phenotype for one sample was DRB1*0701, DRB4*01, DRB5*0101, DRB6*0201, DQB*0602, DQB1*0202. The phenotype of other sample was DRB1*1602, DRB1*1302, DRB3*0301, DRB6*0101, DQB1*0501, DQB1*0502. The first sample has the novel combination of DRB1*0701 with DRB5*0101 and DRB6*0201. The second sample has either DRB6*0101 together with DRB 1*1602 in absence of any DRB5 allele or DRB6*0101 together with DRB1*1302, DRB 3*0301. We postulate that the most likely haplotype in sample #1 is DRB1*0701, DRB5*0101, DRB1*0602 which could have arisen from gene conversion. The most likely haplotype in sample #2, DRB 1*1602, DRB6*0101, DQB1*0502 would have arisen from an homologous recombination event.     

Cardiac arrest: long-term cognitive and imaging analysis

BACKGROUND: Neurological and cognitive sequelae resulting from cardiac arrest (CA), despite their potential personal and social impact, are usually not considered as major outcome measures in long-term analysis of survivors. The aim of this study is to analyze the contribution of neuropsychological testing and cerebral imaging to the development of a long-term classification of neurological impairment. PATIENTS AND METHODS: A total of 19 patients admitted over a 3 years period in an eight-bed intensive care unit of a tertiary care hospital with a diagnosis of CA were alive and attended a 6-month follow-up consultation. Eleven of these patients agreed to participate in this study carried out between 1 and 3 years after CA. Patients were classified using the Cerebral Performance Categories (CPC), neurological examination, detailed cognitive testing and computerized tomography (CT) scan with qualitative and quantitative imaging analysis. RESULTS: Six of the 11 patients had good cerebral performance. Verbal and visuo-spatial short-term memory scores were associated with CPC. All patients with at least moderate cerebral disability had abnormal verbal memory test results compared with only one survivor with CPC 1; visuo-spatial short-term memory was abnormal in four moderately affected survivors and normal in those with CPC 1. The bicaudate ratio evaluated in the CT scan was correlated with the verbal memory score while the III ventricle diameter correlated with the executive functions score, suggesting involvement of different brain areas in these functions. CONCLUSIONS: Neuropsychological and CT scan measurements are proxy measures of long-term impairment of CA survivors, providing a dichotomized global evaluation of CA survivors in close agreement with CPC.