Homozygosity mapping of achromatopsia to chromosome 2 using DNA pooling

Achromatopsia is an autosomal recessive disease of the retina, characterized clinically by an inability to distinguish colors, impaired visual acuity, nystagmus and photophobia. A genome-wide search for linkage was performed using an inbred Jewish kindred from Iran. To facilitate the genome-wide search, we utilized a DNA pooling strategy which takes advantage of the likelihood that the disease in this inbred kindred is inherited by all affected individuals from a common founder. Equal molar amounts of DNA from all affected individuals were pooled and used as the PCR template for short tandem repeat polymorphic markers (STRPs). Pooled DNA from unaffected members of the kindred was used as a control. A reduction in the number of alleles in the affected versus control pool was observed at several loci. Upon genotyping of individual family members, significant linkage was established between the disease phenotype and markers localized on chromosome 2. The highest LOD score observed was 5.4 (theta = 0). When four additional small unrelated families were genotyped, the combined peak LOD score was 8.2. Analysis of recombinant chromosomes revealed that the disease gene lies within a 30 cM interval which spans the centromere. Additional fine-mapping studies identified a region of homozygosity in all affected individuals, narrowing the region to 14 cM. A candidate gene for achromatopsia was excluded from this disease interval by radiation hybrid mapping. Linkage of achromatopsia to chromosome 2 is an essential first step in the identification of the disease-causing gene.      

A potential role for beta- and gamma-crystallins in the vascular remodeling of the eye.

We demonstrate that expression of beta- and gamma-crystallins is associated with intraocular vessels during normal vascular development of the eye and also in the Nuc1 rat, a mutant in which the hyaloid vascular system fails to regress normally. Real-Time RT PCR, Western blot and metabolic labeling studies indicate an increased expression of beta- and gamma-crystallins in Nuc1 retina. The increased expression of crystallins was localized to the astrocytes surrounding the intraocular vessels. A similar pattern of crystallin expression was also observed in the retinal vessels during normal development. Cultured human astrocytes exposed to 3-nitropropionic acid, an established model of neuronal hypoxia, increased VEGF expression, as expected, but also increased expression of crystallins. Our data suggest that crystallins may function together with VEGF during vascular remodeling. Interestingly, in human PFV (persistent fetal vasculature) disease, where the hyaloid vasculature abnormally persists after birth, we show that astrocytes express both VEGF and crystallins.     

Two-dimensional dosimetry in the near field of the model 200 103Pd source for interstitial brachytherapy implants using a thermoluminescent sheet

A large area and highly sensitive thermoluminescent (TL) sheet film was used for two-dimensional dose distribution measurements at millimetre distances from a 103Pd interstitial brachytherapy source. The TL film is made of Teflon homogeneously mixed with small particles of thermoluminescent material (BaSO4: Eu doped). This TL sheet (5 cm x 5 cm) was used to determine the relative dosimetric characteristics (i.e., radial dose function, 2D and 1D anisotropy functions, as defined by the updated AAPM Task Group No 43 report) of the model 200 103Pd source that emits low energy photons (21 keV). The two-dimensional dosimetry data were obtained for distances from the source surface to 15 mm. The radial dose function measured with the TL sheet is in reasonable agreement within 11% with the values recommended in the updated AAPM TG-43 report. All the measured 2D dose distributions showed limited symmetry about the source axes. The differences between the 1D anisotropy function values measured with the TL sheet and the data recommended in the updated AAPM TG-43 report were 10% at 5 mm and 7.5% at 10 mm, respectively, for the model 200 103Pd seed. Our experiments have demonstrated that it is feasible to use the TL sheet as a dosimeter in the determination of the dosimetric characteristics in the immediate vicinity of interstitial brachytherapy sources emitting low energy photons.     

Macrophage-tumour cell interactions: identification of MUC1 on breast cancer cells as a potential counter-receptor for the macrophage-restricted receptor, sialoadhesin

In many carcinomas, infiltrating macrophages are commonly found closely associated with tumour cells but little is known concerning the nature or significance of adhesion molecules involved in these cellular interactions. Here we demonstrate in primary human breast cancers that sialoadhesin (Sn), a macrophage-restricted adhesion molecule, is frequently expressed on infiltrating cells that often make close contact with breast carcinoma cells. To determine whether Sn could act as a specific receptor for ligands on breast cancer cell lines, binding assays were performed with a recombinant form of the protein fused to the Fc portion of human immunoglobulin G1 (IgG1) (Sn-Fc). Sn-Fc was found to bind specifically and in a sialic acid-dependent manner to the breast cancer cell lines MCF-7, T47.D and BT-20 both in solid- and solution-phase binding assays. To investigate the nature of the sialoglycoproteins recognized by Sn on breast cancer cells, MCF-7 cells were labelled with [6-3H]glucosamine. Following precipitation with Sn-Fc, a major band of approximately 240000 MW was revealed, which was shown in reprecipitation and Western blotting experiments to be the epithelial mucin, MUC1.     

Significance of Cryptosporidium in acute diarrhoea in North-Eastern India.

In a hospital-based study, stool samples from 2095 patients of all ages were examined for different fungal, protozoal and bacterial enteropathogens over a period of 2 years (July 1994-June 1996). Cryptosporidium was detected in 151 specimens (7.2%) and was the third commonest pathogen found. The highest prevalence of this organism was in the group aged 16-45 years and during the rainy months (July-Oct.). Diarrhoea caused by the protozoon was of mild to moderate severity and features of dysentery were absent. Amongst other enteropathogens, Candida albicans was the most frequently isolated, followed by enteropathogenic and enterotoxigenic Escherichia coli, Salmonella spp., Campylobacter jejuni, Entamoeba histolytica, Giardia duodenalis (lamblia), Shigella spp., Vibrio cholerae and Aeromonas spp.     

Dose calculation formalisms and consensus dosimetry parameters for intravascular brachytherapy dosimetry: recommendations of the AAPM Therapy Physics Committee Task Group No. 149

Since the publication of AAPM Task Group 60 report in 1999, a considerable amount of dosimetry data for the three coronary brachytherapy systems in use in the United States has been reported. A subgroup, Task Group 149, of the AAPM working group on Special Brachytherapy Modalities (Bruce Thomadsen, Chair) was charged to develop recommendations for dose calculation formalisms and the related consensus dosimetry parameters. The recommendations of this group are presented here. For the Cordis 192Ir and Novoste 90Sr/90Y systems, the original TG-43 formalism in spherical coordinates should be used along with the consensus values of the dose rate constant, geometry function, radial dose function, and anisotropy function for the single seeds. Contributions from the single seeds should be added linearly for the calculation of dose distributions from a source train. For the Guidant 32P wire system, the modified TG-43 formalism in cylindrical coordinates along with the recommended data for the 20 and 27 mm wires should be used. Data tables for the 6, 10, 14, 18, and 22 seed trains of the Cordis system, 30, 40, and 60 mm seed trains of the Novoste system, and the 20 and 27 mm wires of the Guidant system are presented along with our rationale and methodology for selecting the consensus data. Briefly, all available datasets were compared with each other and the consensus dataset was either an average of available data or the one obtained from the most densely populated study; in most cases this was a Monte Carlo calculation.