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Extensive cell movements accompany formation of the otic placode.

During development, the vertebrate inner ear arises from the otic placode, a thickened portion of the ectoderm next to the hindbrain. Here, the first detailed fate maps of this region in the chick embryo are presented. At head process stages, placode precursors are scattered throughout a large region of the embryonic ectoderm, where they intermingle with future neural, neural crest, epidermal, and other placode cells. Within the next few hours, dramatic cell movements shift the future otic placode cells toward the midline and ultimately result in convergence to their final position next to rhombomeres 5-6. Individual cells and small cell groups undergo constant cell rearrangements and appear to sort out from nonotic cells. While the major portion of the otic placode is derived from the nonneural ectoderm, the neural folds also contribute cells to the placode at least until the four-somite stage. Comparison of these fate maps with gene expression patterns at equivalent stages reveals molecular heterogeneity of otic precursor cells in terms of their expression of dlx5, msx1, Six4, and ERNI. Although Pax2 expression coincides with the region where otic precursors are found from stage 8, not all Pax2-positive cells will ultimately contribute to the otic placode.

Animals↗

Using high spectral and spatial resolution bold MRI to choose the optimal oxygenating treatment for individual cancer patients.

We evaluate whether high spectral and spatial resolution (HiSS) BOLD MRI can correctly rank the effects of three tumor-oxygenating treatments on radiosensitivity in BA1112 rhabdomyosarcomas (n = 5). Significant decreases in spectral linewidth predict that treatment with carbogen gas combined with a perfluorocarbon emulsion will increase radiosensitivity more than either treatment alone, which agrees with the known effects of these treatments on hypoxic fraction. High-resolution maps show that tumor response to each treatment is spatially heterogeneous, and that there is a paradoxical response to the treatments in 7-12% of tumor pixels. Because HiSS MRI emphasizes changes in necrotic and/or hemorrhagic regions, it is more sensitive to oxygenation changes compared to conventional MRI. These results demonstrate that HiSS MRI is a practical, noninvasive method that could be used to choose the treatment that maximizes the size and extent of increases in tumor oxygenation for individual patients.

Animals↗

Autosomal dominant polycystic kidney disease: evidence for the existence of a third locus in a Portuguese family.

Autosomal dominant polycystic kidney disease is characterized by clinical and genetic heterogeneity. Two loci implicated in the disease have previously been mapped (PKD1 on chromosome 16 and PKD2 on chromosome 4). By two point and multipoint linkage analysis, negative lod scores have been found for both chromosome 16 and chromosome 4 markers in a large Portuguese family, indicating that a third PKD locus is involved in the development of the disease.

Chromosome Mapping↗

Functional organization in the cat's pulvinar complex.

The responses of 192 single units in the cat's pulvinar-complex, comprising the inferior, medial and lateral pulvinar nuclei, were studied in paralysed cats, lightly anaesthetized with N2O/O2 supplemented with pentobarbitone. About 60% of the cells were visually driven and their receptive fields classified as either diffuse, concentric, movement sensitive, direction sensitive or orientation sensitive. The response fields of such cells were commonly large. Response field maps for the movement and direction sensitive cells formed a heterogenous population with uniform on-off fields to more complex arrangements with on- or off-centres, often with only partial surrounds; other cells responded exclusively to moving stimuli. A dual representation of the visual field was found in the pulvinar-complex corresponding to the striate and tectal recipient zones described anatomically by others. The representation in the striate recipient zone comprised an oblique column running medio-laterally and rostro-caudally through the inferior pulvinar and lateral margin of the medial pulvinar. The peripheral visual field was represented laterally and the vertical meridian medially; the upper visual field was represented dorso-laterally in the medial pulvinar and the lower visual field caudo-ventrally within the inferior pulvinar. That this visuotopic organization corresponded to the striate recipient zone was established by tracing the retrograde transport of HRP. Medial to the striate zone evidence for a second visual field representation was found, apparently more randomly organized than the striate zone, corresponding to the presumed tectal recipient zone. These results support the assertion that cytoarchitectural boundaries do not necessarily delineate functional (visuotopically organized) regions. These observations suggest caution when comparing cytoarchitecturally defined regions between species; rather, 'functionally equivalent' regions should be compared.

Animals↗

Two-dimensional gel electrophoresis of four serum samples from patients with IgD myeloma.

Sera containing IgD paraprotein present problems in identifying patients with monoclonal gammopathies because only a small or even no spike may be present on standard serum protein electrophoresis. We have detected heavy chains of IgD monoclonal protein by means of high resolution two-dimensional gel electrophoresis. Besides clearly identifying delta heavy chains in maps of serum proteins, we also found size and charge heterogeneity of monoclonal immunoglobulins. The results demonstrate the usefulness of two-dimensional gel electrophoresis in the analysis of selected cases of immunoglobulin malignancies.

Aged↗

Absence of linkage of ABO blood group locus to familial tuberous sclerosis.

A mutation leading to tuberous sclerosis was linked to the ABO blood group locus (9q34) on the long arm of chromosome 9. In an effort to confirm this assignment, nine multigenerational families with tuberous sclerosis, comprising 126 sampled individuals, were assessed for linkage of the ABO locus to tuberous sclerosis. Two-point linkage analysis and multilocus linkage analysis were used to evaluate linkage between tuberous sclerosis and the markers ABO, MCT136, and AblK2. Linkage of ABO to tuberous sclerosis was excluded for a distance of 20 centimorgans (cM) encompassing the region of the ABO locus. There was no evidence for genetic heterogeneity within this data set. Using 23 polymorphic markers, exclusion mapping demonstrated only a 1% probability that the tuberous sclerosis locus was on the distal short arm of chromosome 9 and provided no evidence in support of a tuberous sclerosis locus on the remainder of chromosome 9 including the area of the ABO locus.

ABO Blood-Group System↗

Monoclonal antibodies to angiotensin-converting enzyme: a powerful tool for lung and vessel studies.

We obtained a series (12 clones) of hybridomas, which produce monoclonal antibodies (Mab) to 5 different epitopes of the angiotensin-converting enzyme (ACE) molecule. These antibodies may be used to (1) map antigenic structure of ACE, including the study of immunologic heterogeneity of ACE from different organs and tissues; (2) study the immunohistochemical distribution of ACE in human tissues, including the diagnosis of sarcoidosis; (3) develop an ACE immunoassay, and (4) prepare an immunosorbent for large-scale ACE isolation and for ACE-apheresis. One of the antibodies, 9B9, when injected into the circulation of rat and monkey, accumulated with high specificity in the lungs as compared with either normal mouse IgG or other organs and blood. The highly specific and nontoxic accumulation of Mab 9B9 suggests that it also may be used for gamma scintigraphy visualization of the pulmonary vascular bed, detection of lung injury and as a vector for targeted drug delivery to the lung.

Animals↗

Variable and constant regions in African swine fever virus DNA.

An analysis of the SalI restriction pattern of African swine fever virus DNA showed that the SalI recognition sites did not change after more than 100 virus passages in porcine macrophages. The virus strain BA71V, obtained from the virus isolate BA71 by adaptation to grow in VERO cells, differed from the nonadapted virus in two deletions with a length of 2.5 and 7 kb located close to the DNA ends. A restriction analysis of several virus clones obtained from a naturally infected pig revealed length heterogeneity in both variable regions. A comparison of SalI restriction maps from 23 African swine fever virus field isolates (8 African, 11 European, and 4 American) has shown that the virus genome consists of a central region with a constant length of about 125 kb and two variable regions located close to the DNA ends with a length of 38-47 kb for the left DNA end, and 13-16 kb for the right DNA end. The total length of ASF virus DNA varied between 178 (BA71) and 189 (MOZ64) kb. The 23 African swine fever virus isolates were classified into five groups, according to the arrangement of the SalI sites in the central region. Four groups contained only African isolates, whereas all the European and American isolates belonged to the same group. This distribution of isolates suggests that all non-African virus field isolates have a common origin.

Africa↗

Fluid velocity imaging of reservoir core samples.

Experimental definition of fluid flow in heterogeneous reservoir rock requires three dimensional imaging techniques. Fluid velocity mapping at a steady-state flow condition plus oil/water saturation mapping using MRI provides a high resolution view of the process.

Kerosene↗

Intracoronary thermography for detection of high-risk vulnerable plaques.

Up to two-thirds of acute myocardial infarctions develop at sites of culprit lesions without a significant stenosis. New imaging techniques are needed to identify those lesions with an increased risk of developing an acute complication in the near future. Inflammation is a hallmark feature of these vulnerable/high-risk plaques. We have shown that inflamed atherosclerotic plaques are hot and their surface temperature correlates with an increased number of macrophages and decreased fibrous-cap thickness. Multiple animal and human experiments have shown that temperature heterogeneity correlates with arterial inflammation in vivo. Several coronary temperature mapping catheters are currently being developed and studied. These thermography methods can be used in the future to detect vulnerable plaques, potentially to determine patients' prognosis, and to study the plaque-stabilizing effects of different medications.

Animals↗

A posttranscriptional regulator of Kaposi's sarcoma-associated herpesvirus interacts with RNA-binding protein PCBP1 and controls gene expression through the IRES.

Kaposi's sarcoma-associated herpesvirus (KSHV; also known as human herpesvirus 8, HHV-8) belongs to the gamma-herpesvirus subfamily. The KSHV ORF57 gene is thought to be a homolog of posttranscriptional regulators that are conserved in the herpesvirus family and are essential for replication. We generated specific monoclonal antibodies (mAbs) against the ORF57 protein that detected the 51-kDa protein expressed in the nucleus of KSHV-infected cells. We also found that the ORF57 protein interacted with poly(rC)-binding protein 1 (PCBP1), a cellular RNA-binding, posttranscriptional regulator. ORF57's interaction with PCBP1 enhanced the activity of not only poliovirus internal ribosome-entry site (IRES)-dependent translation but also X-linked inhibitor of apoptosis (XIAP) and KSHV vFLIP IRES. Actually, when ORF57 expression was induced by the expression of replication and transcription activator (RTA) in KSHV-infected cells, the expression of XIAP was enhanced. These results suggest that ORF57 binds to PCBP1 as a functional partner for posttranscriptional regulation and is involved in the regulation of the expression of both cellular and viral genes through IRESs.

Base Sequence↗

The genetics of gene expression and gene mapping.

A recent publication has shown that a significant portion of gene expression levels are under genetic control in different organisms, that there are hotspot regions in the genome that control the expression of many other genes, and how gene expression data can be used to localize genes that affect clinical traits.

Adipose Tissue↗

A two-locus model for non-syndromic congenital dysplasia of the hip (CDH).

Complex segregation analysis was conducted in a sample of 171 pedigrees collected through probands affected by non-syndromic dysplasia of the hip (CDH) treated in Ferrara's CDH Centre in the period 1991-6. The analysis favoured a two-locus model, in which the accepted segregation model at the major locus was compatible with recessive transmission, with a gene frequency of the deleterious allele of around 0.20. For the other locus, among the Mendelian hypotheses tested, the recessive model turned out to be the most parsimonious. When ultrasonographic level was examined as an indication of severity, a significant improvement in the fit of the model was observed, giving clear evidence that information on the severity of CDH is important for a better definition of the hereditary transmission model.

Child↗

Genetic susceptibility to prostate cancer: a review.

A genetic component in prostate cancer has been recognized since decades. Through numerous epidemiological and molecular biological studies, much evidence has accumulated in favor of a significant but heterogeneous hereditary component in prostate cancer (PCa) susceptibility. Since the mapping of a high-penetrant PCa susceptibility locus at 1q24-25, much attention has been paid to the identification of PCa susceptibility genes. So far, seven loci have been mapped, and at three of these loci, genes have been cloned and mutations identified. Yet their role in hereditary and sporadic disease is still under debate and probably very modest. Although research on hereditary prostate cancer has improved our knowledge of the genetic etiology of the disease, still a lot of questions remain unanswered. Here, we aim to review the genetic epidemiological and molecular biological research in the field of hereditary prostate cancer and the problems that are encountered with this research.

Chromosomes, Human, Pair 1↗

Loss-of-function mutations in a calcium-channel alpha1-subunit gene in Xp11.23 cause incomplete X-linked congenital stationary night blindness.

X-linked congenital stationary night blindness (CSNB) is a recessive non-progressive retinal disorder characterized by night blindness, decreased visual acuity, myopia, nystagmus and strabismus. Two distinct clinical entities of X-linked CSNB have been proposed. Patients with complete CSNB show moderate to severe myopia, undetectable rod function and a normal cone response, whereas patients with incomplete CSNB show moderate myopia to hyperopia and subnormal but measurable rod and cone function. The electrophysiological and psychophysical features of these clinical entities suggest a defect in retinal neurotransmission. The apparent clinical heterogeneity in X-linked CSNB reflects the recently described genetic heterogeneity in which the locus for complete CSNB (CSNB1) was mapped to Xp11.4, and the locus for incomplete CSNB (CSNB2) was refined within Xp11.23 (ref. 5). A novel retina-specific gene mapping to the CSNB2 minimal region was characterized and found to have similarity to voltage-gated L-type calcium channel alpha1-subunit genes. Mutation analysis of this new alpha1-subunit gene, CACNA1F, in 20 families with incomplete CSNB revealed six different mutations that are all predicted to cause premature protein truncation. These findings establish that loss-of-function mutations in CACNA1F cause incomplete CSNB, making this disorder an example of a human channelopathy of the retina.

Amino Acid Sequence↗

Isolation from human chronic-lymphocytic-leukaemia cells of membrane glycoproteins associated with Fc-receptor functions. Physical parameters and production of polyclonal antibodies.

Two membrane glycoproteins that bound immune complexes and inhibited Fc-receptor- (FcR-)mediated functions in vitro were purified from human FcR+ chronic-lymphocytic-leukaemia cells. A multi-step purification was developed, consisting essentially in: (i) Tween 40 extraction of crude cell membranes; (ii) solubilization of membrane fragments by Renex-30; (iii) isolation of glycoproteins by affinity chromatography on Lens culinaris haemagglutinin-Sepharose; (iv) papain treatment of the eluted glycoproteins followed by gel-filtration chromatography; (v) purification by polyacrylamide-gel electrophoresis of two molecular species from the protein-size fraction enriched for immune-complex-binding activity. The two electrophoretically isolated components displayed apparent molecular masses of 70 and 45 kDa by SDS/polyacrylamide-gel electrophoresis and restricted charge heterogeneity by two-dimensional analysis. Two-dimensional peptide mapping revealed the presence of many peptides in common between the two proteins and the absence of a number of peptides in the 45 kDa component. These two polypeptides were used as immunogens to produce polyclonal antibodies that cross-reacted with both proteins and specifically inhibited FcR-mediated reactions in vitro. Furthermore, FcR-related components from detergent-extracted lysates of the human K562 and U937 cell lines or human placental membranes were revealed by the putative anti-FcR antibodies adsorbed on Protein A-Sepharose.

Antigen-Antibody Complex↗

Renaturation of complementary DNA strands mediated by purified mammalian heterogeneous nuclear ribonucleoprotein A1 protein: implications for a mechanism for rapid molecular assembly.

Purified heterogeneous nuclear ribonucleoprotein (hnRNP) A1 protein, which is found in vivo associated with heterogeneous nuclear RNA (hnRNA), promotes the rapid renaturation of nucleic acid strands. Maximal renaturation activity requires the glycine-rich carboxyl-terminal one-third of the protein, although the amino-terminal two-thirds also has activity. The A1-mediated reaction is second-order with respect to complementary DNA concentration, and the renaturation rate constant at 37 degrees C with A1 is about 3000-fold greater than in the absence of the protein. At 60 degrees C, the A1-mediated renaturation rate is even faster, and is about 300-fold greater than protein-free reactions carried out at 68 degrees C in 1 M NaCl. Provided that sufficient A1 protein is present to coat all strands in solution, the presence of nonhomologous, single-stranded DNA does not significantly inhibit the reaction. Moreover, renaturation of short strands to their complement contained in very long strands is nearly as efficient as between two short strands. These results indicate that A1 may be useful for procedures that rely on nucleic acid renaturation. We propose that A1 promotes rapid renaturation primarily by reducing the entropic barrier of bimolecular strand association through relatively transient interactions between A1-coated strands. Such interactions, mediated by flexible repeating domains, may act generally to increase the association kinetics of highly specific molecular assemblies in processes such as RNA maturation, transcription, translation, and transport.

Coliphages↗

Mathematics-assisted mapping in analysis of medical disease.

Genetic mapping in analysis of medical disease is performed under several assumptions and (experimental) conditions, which are made about the data in general and the disease in particular. Here we discuss these conditions, what they mean, and what kind of deleterious effects they might have on the analysis. We also illustrate how to proceed and what kind of possibilities the statistical analysis may provide to medical scientists.

Chromosome Mapping↗