A mutant wfs1 zebrafish model of Wolfram syndrome manifesting visual dysfunction and developmental delay.

Wolfram syndrome (WS) is an ultra-rare progressive neurodegenerative disorder defined by early-onset diabetes mellitus and optic atrophy. The majority of patients harbour recessive mutations in the WFS1 gene, which encodes for Wolframin, a transmembrane endoplasmic reticulum protein. There is limited availability of human ocular and brain tissues, and there are few animal models for WS that replicate the neuropathology and clinical phenotype seen in this disorder. We, therefore, characterised two wfs1 zebrafish knockout models harbouring nonsense wfs1a and wfs1b mutations. Both homozygous mutant wfs1a-/- and wfs1b-/- embryos showed significant morphological abnormalities in early development. The wfs1b-/- zebrafish exhibited a more pronounced neurodegenerative phenotype with delayed neuronal development, progressive loss of retinal ganglion cells and clear evidence of visual dysfunction on functional testing. At 12 months of age, wfs1b-/- zebrafish had a significantly lower RGC density per 100 µm2 (mean ± standard deviation; 19 ± 1.7) compared with wild-type (WT) zebrafish (25 ± 2.3, p < 0.001). The optokinetic response for wfs1b-/- zebrafish was significantly reduced at 8 and 16 rpm testing speeds at both 4 and 12 months of age compared with WT zebrafish. An upregulation of the unfolded protein response was observed in mutant zebrafish indicative of increased endoplasmic reticulum stress. Mutant wfs1b-/- zebrafish exhibit some of the key features seen in patients with WS, providing a versatile and cost-effective in vivo model that can be used to further investigate the underlying pathophysiology of WS and potential therapeutic interventions.

Transcriptome profiling of zebrafish optic fissure fusion.

Incomplete fusion of the optic fissure leads to ocular coloboma, a congenital eye defect that affects up to 7.5 per 10,000 births and accounts for up to 10 percent of childhood blindness. The molecular and cellular mechanisms that facilitate optic fissure fusion remain elusive. We have profiled global gene expression during optic fissure morphogenesis by transcriptome analysis of tissue dissected from the margins of the zebrafish optic fissure and the opposing dorsal retina before (32 hours post fertilisation, hpf), during (48 hpf) and after (56 hpf) optic fissure fusion. Differential expression analysis between optic fissure and dorsal retinal tissue resulted in the detection of several known and novel developmental genes. The expression of selected genes was validated by qRT-PCR analysis and localisation investigated using in situ hybridisation. We discuss significantly overrepresented functional ontology categories in the context of optic fissure morphogenesis and highlight interesting transcripts from hierarchical clustering for subsequent analysis. We have identified netrin1a (ntn1a) as highly differentially expressed across optic fissure fusion, with a resultant ocular coloboma phenotype following morpholino antisense translation-blocking knockdown and downstream disruption of atoh7 expression. To support the identification of candidate genes in human studies, we have generated an online open-access resource for fast and simple quantitative querying of the gene expression data. Our study represents the first comprehensive analysis of the zebrafish optic fissure transcriptome and provides a valuable resource to facilitate our understanding of the complex aetiology of ocular coloboma.

[The diagnostic significance of biochemical markers.]

INTRODUCTION: The modern diagnostic of neuroendocrine tumors is based on analysis of compounds produced by tumor cells (peptides, amines, hormones). PURPOSE OF STUDY: To evaluate diagnostic effectiveness of Chromogranin A, serotonin and its metabolite 5-hydroxyindolilacetic acid as biochemical markers of neuroendocrine tumors as biochemical markers of neuroendocrine tumors. MATERIAL AND METHODS: The detection of Chromogranin A and serotonin in blood serum and 5-hydroxyindolilacetic acid in day urine was applied to 330 patients with neuroendocrine tumors of lungs, pancreas, stomach, small and large intestines and also to 115 healthy males and females. The detection was implemented by enzyme-linked immunosorbent assay based on testsystems "Chromogranin A ELISA Kit" (Dako A/S, Denmark), "Serotonin ELISA" and 5-HIAA ELISA (IBL International GMBH, Germany). RESULTS AND DISCUSSION: The levels of Chromogranin A at all localizations of neuroendocrine tumors reliably (p<0.000001) exceeded corresponding control index. In case of serotonin and 5-hydroxyindolilacetic acid reliable differences were established in all groups except neuroendocrine tumors of stomach. The dependence is established between secretion of markers from prevalence and activity of neuroendocrine tumors. The corresponding levels were higher in patients with metastases in liver and under carcinoid syndrome as compared with patients without corresponding clinical manifestations. The evaluation of diagnostic significance of Chromogranin A, serotonin and 5-hydroxyindolilacetic acid was applied considering threshold levels calculated according results of their detection in control group (33 ng/ml, 320 ng/ml and 60 mkmol/day correspondingly). The high diagnostic sensitivity of Chromogranin A was demonstrated that amounted to 80.6% at specificity 98.5% on the whole in group of patients with neuroendocrine tumors. The serotonin and 5-hydroxyindolilacetic acid manifested comparable diagnostic sensitivity only in patients with carcinoid syndrome (72.5 and 60.3%). CONCLUSION: The obtained data substantiate high effectiveness of Chromogranin A as a marker of neuroendocrine tumors. The detection of this marker contributes into enhancement of accuracy of diagnostic and evaluation of prevalence of tumors of neuroendocrine nature. The serotonin and 5-hydroxyindolilacetic acid are markers of carcinoid syndrome.

Biochemical markers of brain tumours.

The paper presents the results of neurospecific proteins S-100 and glial fibrillary acidic protein (GFAP) determination in blood serum samples of 145 neuro-oncology patients and 69 healthy people. The significant elevation of S-100 and GFAP was revealed in glioblastoma (G IV) patients compare to the patients with anaplastic astrocytoma (G III), benign meningioma (GI), celebral metastasis and healthy controls. The concentration of S-100 in blood serum of patients with anaplastic astrocytoma, benign meningioma, and celebral metastasis did not significantly differ among themselves, and in relation to the control group there was a significantly increase only in patients with cerebral metastasis. GFAP was characterized by high frequency of its detection in patients with glioblastoma (83%) compare to other groups of patients and healthy donors, in which it was practically undetectable. These data suggest the possibility of using GFAP as a marker of glioblastoma and S-100- as an additional biochemical criteria of cerebral lesions in oncology patients.

Neurospecific proteins in the serum of patients with brain tumors.

Neurospecific proteins S-100 and GFAP were measured in the serum of 145 patients with neural tumors and 69 healthy individuals. In patients with glyoblastomas, the concentrations of S-100 and GFAP were significantly higher than in patients with anaplastic astrocytomas, benign meningiomas, and brain metastases and in healthy individuals. Serum S-100 concentrations in patients with anaplastic astrocytomas, benign meningiomas, and brain metastases were similar; significant difference from the control was found only for patients with cerebral metastases. A specific feature of GFAP was high incidence of its detection in patients with glioblastomas (83%) compared to other groups of patients with neural tumors and healthy volunteers who demonstrated practically zero level of this protein. These findings attest to the possibility of using S-100 as an additional biochemical criterion of brain involvement in tumor patients and GFAP as a glioblastoma marker.

Biochemical parameters in the diagnosis and monitoring of neurotoxicity of antitumor cytostatics.

The significance of neurospecific proteins in the diagnosis of neurotoxicity in patients with breast, lung, testicular, and ovarian cancer treated by taxane and cisplatin drugs was evaluated. The most pronounced increase in the content of these proteins and titers of autoantibodies to these proteins was observed in patients with clinical manifestations of neurotoxicity induced by cytostatics. A strong correlation was found between the concentration of myelin basic protein and cumulative dose of the drug (R=0.922; p<0.0001). These data suggest that myelin basic protein and gliofibrillar acid protein can be used as markers in the diagnosis and monitoring of antitumor drug neurotoxicity.

Mood and cognition in pregnant workers.

Thirty-eight working women, in their late second trimester or early third trimester of pregnancy, volunteered to fill out the Cognitive Failures Questionnaire and the UWIST Mood Adjective Checklist. In addition they provided information on the number of children they already had, the nature of their employment and their education level. The latter information was used to select a match for each pregnant worker from a larger sample of non-pregnant female workers. The results revealed no differences in the level of cognitive failures between the two groups. However, CFQ score was a better predictor of mood in the pregnant subjects, suggesting mood changes may be more closely related to perceived cognitive competency during pregnancy. The implications of this are discussed. With respect to mood sub-scales, non-pregnant workers scored higher on Energetic Arousal than pregnant workers. There were no differences on other mood sub-scales. It is argued that the cognitive efficiency of workers is not compromised by pregnancy but steps should be taken to ensure that work load is adjusted to take account of the self-reported reduced arousal that may arise.

Human microvascular endothelial cell-extracellular matrix interaction in cellular growth state determination.

We introduce two methods, both of which are based on cellular-extracellular matrix interaction, which will facilitate the study of human microvascular endothelial cells. One method describes the means to obtain a G1 population baseline in human microvascular endothelial cells. Because of the contribution of the extracellular matrix in endothelial cell growth, synchronization in G1 was possible only after the incorporation of angiostatic levels of heparin and hydrocortisone into the extracellular matrix. In the second method, we demonstrate that selective perturbation of human microvascular endothelial cell-extracellular matrix interactions results in the induction of a transitional growth state, between proliferative and differentiated growth states, in human microvascular endothelial cells. In the functional, microtubule formation assays, transitional growth state endothelial cells display rates that are indermediate between those obtained from differentiated and proliferative endothelial cells. Our results demonstrate the importance of the human microvascular endothelial cell-extracellular matrix interaction in the determination of cellular growth state. Our findings also imply that responsiveness of microvascular endothelial cells to their cellular-extracellular matrix environs is highest during the differentiated growth state.

Characteristics of visual interference with visuospatial working memory.

Recent discussions of visuospatial working memory have suggested that this subsystem may incorporate a visual buffer which holds visuospatial information relatively passively. Empirical investigations of visual interference with information held within a visuospatial subsystem have yielded somewhat equivocal results. Nonetheless, evidence from Logie (1986) has indicated that visuospatial processing can be disrupted by passive exposure to irrelevant visual material in a manner analogous to the disruption of serial verbal recall by exposure to irrelevant speech. This paper reports two experiments which explore whether such irrelevant visual input is disruptive to storage of imaginal information in a primarily spatial task--the Brooks spatial matrix task. Experiment 1 shows that exposure to irrelevant visual input during encoding selectively disrupts performance on a spatial, but not a verbal, version of the task. The extent of such disruption is shown to be independent of the visual complexity of the material, its similarity to the to-be-remembered information, or a change in state, with a static white square pattern yielding equivalent disruption to that produced by changing matrix patterns. The second experiment indicates that this pattern of effects is robust, and that such disruption is evident at an equivalent level when the visual material is present only during a 20-second retention interval. These results are interpreted as evidence of obligatory access of external visual material to a passive visual buffer. Implications for the nature of a visuospatial subsystem in working memory are discussed.

In utero exposure to diagnostic ultrasound: a 6-year follow-up.

A 6-year follow-up study of height and weight was conducted between 1975 and 1983 in a total of 149 sibling pairs of the same sex, one of whom had been exposed to diagnostic ultrasound in utero. The data were analyzed for the entire sample group, as well as for subgroups defined by sex, gestational age at delivery, birth order, and gestational age at time of first exposure to ultrasound. No statistically significant differences of head circumference at birth or of height and weight between birth and 6 years of age were found between ultrasound-exposed and unexposed siblings. In our sample population, exposure of fetuses to ultrasound did not significantly affect growth in childhood up to 6 years of age.

Clinical dosimetry for radiotherapy to the breast based on imaging with the prototype Royal Marsden Hospital CT simulator.

The result of conservative management of breast cancer is strongly dependent on the homogeneity of the dose delivered a schedule of post-surgical radiotherapy. In addition to improvements in local control, late normal-tissue effects should be minimised by achieving a good dose homogeneity. The Royal Marsden Hospital prototype CT simulator has been used to image patients in the treatment position. With the CT data incorporated into the planning process a quantitative measure of the dose homogeneity was made. There are strong indications that unless tissue compensators are used and/or conformation therapy is performed, the dose inhomogeneity in a widely used treatment geometry is too large to be clinically acceptable.

Benign muscular dystrophy with contractures: a new syndrome?

Three patients are described with muscular dystrophy and contractures. Although this disorder bears similarities to Emery-Dreifuss disease and variants previously described, absence of cardiomyopathy is a distinguishing feature. Electrodiagnostic testing and muscle biopsy are consistent with a myopathy. An autosomal dominant pattern of inheritance is suspected, but the possibility of a Y-to-Y transmission cannot be completely excluded.