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Functional replacement of Wheat streak mosaic virus HC-Pro with the corresponding cistron from a diverse array of viruses in the family Potyviridae.

Helper component-proteinase (HC-Pro) of Wheat streak mosaic virus strain Sidney 81 (WSMV-Sidney 81) was systematically replaced with the corresponding cistron derived from four strains of WSMV (Type, TK1, CZ, and El Batán 3), the tritimovirus Oat necrotic mottle virus (ONMV), the rymoviruses Agropyron mosaic virus (AgMV) and Hordeum mosaic virus (HoMV), or the potyviruses Tobacco etch virus (TEV) and Turnip mosaic virus (TuMV). These HC-Pro proteins varied in amino acid sequence identity shared with HC-Pro of WSMV-Sidney 81 from high (strains of WSMV at approximately 86-99%) to moderate (ONMV at 70%) to low (rymoviruses and potyviruses at approximately 15-17%). Surprisingly, all chimeric viral genomes examined were capable of systemic infection of wheat upon inoculation with RNA transcripts produced in vitro. HC-Pro replacements derived from tritimoviruses did not alter host range relative to WSMV-Sidney 81, as each of these chimeric viruses was able to systemically infect wheat, oat, and corn line SDp2. These results indicate that differences in host range among tritimoviruses, including the inability of ONMV to infect wheat or the inability of WSMV strains Type and El Batán 3 to infect SDp2 corn, are not determined by HC-Pro. In contrast, all chimeric viruses bearing HC-Pro replacements derived from rymoviruses or potyviruses were unable to infect SDp2 corn and oat. Collectively, these results indicate that HC-Pro from distantly related virus species of the family Potyviridae are competent to provide WSMV-Sidney 81 with all functions necessary for infection of a permissive host (wheat) and that virus-host interactions required for systemic infection of oat and SDp2 corn are more stringent. Changes in symptom severity or mechanical transmission efficiency observed for some chimeric viruses further suggest that HC-Pro affects virulence in WSMV.

Amino Acid Sequence↗

Altering AAV tropism with mosaic viral capsids.

Over the past decade, AAV-based vectors have emerged as promising candidates for gene therapeutic applications. Despite the broad tropism of the first eight serotypes identified, certain cell types are refractory to transduction with AAV-based vectors. Furthermore, for certain applications the targeting of specific cell types is desirable. To improve on present methods to alter AAV2 tropism, we take advantage of AAV2 mosaics. Here, we show that AAV2 mosaics have improved infectivity compared with all-mutant virions. Using an AAV2 mutant that contains the immunoglobulin-binding Z34C fragment of protein A, we demonstrate the utility of AAV2 mosaics to alter AAV2 tropism. This system allows us to transduce selectively and efficiently MO7e and Jurkat cells. The use of AAV2 mosaics with a protein A fragment inserted into their capsid, together with targeting antibodies, is a versatile method that allows the specific transduction of a wide array of cell types.

Capsid↗

Regeneration of the dopamine-cell mosaic in the retina of the goldfish.

A fundamental anatomical feature of retinal neurons is that they form planar mosaics. Each mosaic can be described by its density, pattern, and regularity (non-randomness). As part of ongoing studies to quantitatively describe the anatomy of regenerated retina in the goldfish, we determined the planimetric density and regularity of the mosaic of dopaminergic interplexiform cells in patches of regenerated retina and compared this to the mosaic generated de novo. In addition, we selectively ablated dopaminergic neurons with the neurotoxin 6-hydroxydopamine (6-OHDA) before inducing local regeneration and determined whether or not the absence of the extant dopaminergic neurons modulated the planimetric density or number of regenerated ones. The results showed that dopaminergic neurons are regenerated at higher planimetric densities and in less orderly arrays than normal. Furthermore, there was no statistical difference in the density or number of regenerated cells in normal retinas and retinas treated with 6-OHDA.

Animals↗

The cone photoreceptor mosaic of the green sunfish, Lepomis cyanellus.

Recent empirical and theoretical evidence has implicated the geometrical birefringence of the double cones of the green sunfish (Lepomis cyanellus) as the biophysical basis of this vertebrate's sensitivity to polarized light. Because of the intimate link between the organization of the cone-photoreceptor mosaic and the psychophysical details of polarization sensitivity, we have examined the structural features of the green sunfish cone-photoreceptor mosaic, in particular the orientation of the elliptical cross sections of the double cones. Our primary observations are that (1) the arrangement of the cone-photoreceptor mosaic is constant across the retina (with two regional exceptions), with double cones arranged in a rhombic mosaic and aligned roughly +/- 45 deg to the nearest retinal margin; (2) the double-cone/single-cone ratio is everywhere the same; (3) cone density is inhomogeneous across the retina, with the highest densities in the temporal hemiretina. These results are discussed as they relate to the animal's retinal growth and visual mechanisms, particularly the sensitivity to polarized light.

Animals↗

Mosaic properties of midget and parasol ganglion cells in the marmoset retina.

We measured mosaic properties of midget and parasol ganglion cells in the retina of a New World monkey, the common marmoset Callithrix jacchus . We addressed the functional specialization of these populations for color and spatial vision, by comparing the mosaic of ganglion cells in dichromatic ("red-green color blind") and trichromatic marmosets. Ganglion cells were labelled by photolytic amplification of retrograde marker ("photofilling") following injections into the lateral geniculate nucleus, or by intracellular injection in an in vitro retinal preparation. The dendritic-field size, shape, and overlap of neighboring cells were measured. We show that in marmosets, both midget and parasol cells exhibit a radial bias, so that the long axis of the dendritic field points towards the fovea. The radial bias is similar for parasol cells and midget cells, despite the fact that midget cell dendritic fields are more elongated than are those of parasol cells. The dendritic fields of midget ganglion cells from the same (ON or OFF) response-type array show very little overlap, consistent with the low coverage of the midget mosaic in humans. No large differences in radial bias, or overlap, were seen on comparing retinae from dichromatic and trichromatic animals. These data suggest that radial bias in ganglion cell populations is a consistent feature of the primate retina. Furthermore, they suggest that the mosaic properties of the midget cell population are associated with high spatial resolution rather than being specifically associated with trichromatic color vision.

Animals↗

Developmental improvement in the regularity and packing of mouse horizontal cells: implications for mechanisms underlying mosaic pattern formation.

The present investigation has sought to determine whether the population of retinal horizontal cells undergoes an increase in the precision of its mosaic patterning during postnatal development, and if so, whether this increase is compatible with three different mechanistic accounts of retinal mosaic formation. Horizontal cells were labeled with antibodies to neurofilaments or calbindin at different developmental stages, and then visualized in retinal wholemounts. Multiple fields were sampled from each retina to determine horizontal cell density, while the X-Y coordinates of each cell in a field were determined. An estimate of total horizontal cell number was calculated for each retina, while the Voronoi domain regularity index and the packing factor were computed for each field. Two strains of mice showing a two-fold difference in the size of their horizontal cell population in maturity were sampled, C57BL/6J and A/J. Horizontal cell number in C57BL/6J was approximately twice that observed in A/J at all postnatal stages, with neither strain showing an effect of age on horizontal cell number. In both strains, however, the Voronoi domain regularity index and the packing factor were significantly lower at P-1 relative to later developmental stages. These results show that accounts of mosaic formation proposing the selective death of irregularly positioned cells, or the periodic occurrence of fate-determining events, are insufficient to establish the final patterning achieved by horizontal cells. Rather, they support the hypothesis that tangential dispersion enhances mosaic patterning during postnatal development.

Aging↗

3'-Terminal sequence analysis of the RNA genome of the Indian isolate of cardamom mosaic virus: a new member of genus Macluravirus of potyviridae.

Cardamom mosaic virus, a possible member of the family Potyviridae has been associated with the mosaic disease (Katte disease) of small cardamom in India. A virus isolated from the symptomatic cardamom leaves was positive in ELISA only with antiserum to the Guatemalan isolate of cardamom mosaic virus and not with a number of other potyviruses. The size of the viral RNA (8.5 kb) and the molecular weight of the coat protein (CP) (38 kDa) were determined. A 1.8-kb product containing the partial nuclear inclusion body (NIb) gene, the entire coat protein gene and the 3' untranslated region (UTR) was amplified by reverse transcription (RT) and polymerase chain reaction (PCR), cloned and sequenced. The viral origin of the clone was confirmed by Northern hybridization with viral RNA. The experimentally determined N-terminal sequence of the CP matched with the deduced amino acid sequence of the CP gene. Sequence analysis of the clone suggests that the cardamom mosaic virus is a member of the Maclurvirus genus of the family Potyviridae.

3' Untranslated Regions↗

Effects of scale and logging on landscape structure in a forest mosaic.

Landscape structure in a forest mosaic changes with spatial scale (i.e. spatial extent) and thresholds may occur where structure changes markedly. Forest management alters landscape structure and may affect the intensity and location of thresholds. Our purpose was to examine landscape structure at different scales to determine thresholds where landscape structure changes markedly in managed forest mosaics of the Appalachian Mountains in the eastern United States. We also investigated how logging influences landscape structure and whether these management activities change threshold values. Using threshold and autocorrelation analyses, we found that thresholds in landscape indices exist at 400, 500, and 800 m intervals from the outer edge of management units in our study region. For landscape indices that consider all landcover categories, such as dominance and contagion, landscape structure and thresholds did not change after logging occurred. Measurements for these overall landscape indices were strongly influenced by midsuccessional deciduous forest, the most common landcover category in the landscape. When restricting analyses for mean patch size and percent cover to individual forest types, thresholds for early-successional forests changed after logging. However, logging changed the landscape structure at small spatial scale, but did not alter the structure of the entire forest mosaic. Previous forest management may already have increased the heterogeneity of the landscape beyond the point where additional small cuts alter the overall structure of the forest. Because measurements for landscape indices yield very different results at different spatial scales, it is important first to identify thresholds in order to determine the appropriate scales for landscape ecological studies. We found that threshold and autocorrelation analyses were simple but powerful tools for the detection of appropriate scales in the managed forest mosaic under study.

Environmental Monitoring↗

Corneal mosaic patterns--morphology and epidemiology.

The corneas of 1485 consecutive patients were examined for the presence of a mosaic pattern. One hundred and ninety-nine peripheral mosaic patterns, ten posterior crocodile shagreens and two cloudy dystrophies were found. Peripheral mosaic pattern is age related and distinct from corneal arcus. Posterior crocodile shagreen is also age related and in one case was found to be unilateral. The morphology of the mosaic patterns is discussed.

Adolescent↗

Binding and translation of turnip-yellow-mosaic-virus ribonucleic acid by skeletal-muscle ribosomes from normal and diabetic rats.

Ribosomes from skeletal muscle of diabetic rats were less active than normal ribosomes in protein synthesis directed by turnip-yellow-mosaic-virus RNA. The proportion of ribosomes from muscle of diabetic rats capable of binding turnip-yellow-mosaic-virus RNA was greater than normal, but there was no difference in the equilibrium constants for the binding reaction. The turnip-yellow-mosaic-virus RNA was bound preferentially to the small (40S) ribosomal subunit, whereas the decrease due to diabetes in its translation was associated with the large (60S) subunit. Thus the diminished capacity of ribosomes from muscle of diabetic rats to translate turnip-yellow-mosaic-virus RNA was not the result of decreased binding of the template.

Animals↗

Modelling the mosaic organization of rod and cone photoreceptors with a minimal-spacing rule.

The mosaic of photoreceptors is regarded as a prime example of the precise control of cellular positioning in the vertebrate nervous system. This study was undertaken with the idea that understanding the intrinsic geometrical features of photoreceptor mosaics is a necessary step to unveil the biological mechanisms governing their formation. We show in the retina of the ground squirrel that the arrays of both the rods and S cones are non-random, but that nothing more than a simple minimal-spacing rule constraining receptor positioning is sufficient to account for the spatial organization of both mosaics. The size of this 'exclusion zone' is an intrinsic characteristic of each cell type, and it is simply the difference in the size of this domain that accounts for the regularity of the S cone array and the irregularity of the rod array at identical density. Consequently, regularity in receptor mosaics is produced by two independent biological events, one embodying the exclusion zone, and another specifying the local density of a given receptor type.

Animals↗

Mosaic mice with teratocarcinoma-derived mutant cells deficient in hypoxanthine phosphoribosyltransferase.

Mutagenized stem cells of a cultured mouse teratocarcinoma cell line were selected for resistance to the purine base analog 6-thioguanine. Cells of a resistant clone were completely deficient in activity of the enzyme hypoxanthine phosphoribosyltransferase (HPRT, IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8), the same X-linked lesion as occurs in human Lesch-Nyhan disease. After microinjection into blastocysts of another genetic strain, the previously malignant cells successfully participated in normal embryogenesis and tumor-free, viable mosaic mice were obtained. Cells of tumor lineage were identified by strain markers in virtually all tissues of some individuals. Mature function of those cells was evident from their tissue-specific products (e.g., melanins, liver proteins). These mutagenized teratocarcinoma cells are therefore developmentally totipotent. Retention of the severe HPRT deficiency in the differentiated state was documented in extracts of mosaic tissues by depressed specific activity of the enzyme, and also by presence of unlabeled clones in autoradiographs of explanted cells incubated in [(3)H]hypoxanthine. Some mosaic individuals had mutant-strain cells in only one or a few tissues. Such animals may provide unique opportunities to identify the tissue sources of particular aspects of the complex disease syndrome. The tissue distribution of HPRT-deficient cells suggests that selection against them is particularly strong in blood of the mosaic mice, as is already known to be the case in human heterozygotes. This phenotypic parallelism supports the expectation that afflicted F(1) male mice that might be obtained from mutant germ cells can serve as a model of the human disease.

Animals↗

Replication of pea enation mosaic virus RNA in isolated pea nuclei.

Isolated nuclei from healthy pea plants were primed with pea enation mosaic virus (PEMV), southern bean mosaic virus (SBMV), radish mosaic virus (RdMV), tobacco mosaic virus (TMV), PEMV RNA, SBMV RNA, RdMV RNA, or TMV RNA. RNA replication occurred only with PEMV RNA and not with intact PEMV or any of the other viruses or RNAs, as judged by ensuing actinomycin D-insensitive polymerase activity. Molecular hybridization experiments showed that some of the product of the polymerase was PEMV-specific (-)RNA. The substrate and ionic requirements of this polymerase were the same as those for the RNA-dependent RNA polymerase present in nuclei isolated from PEMV-infected pea plants. No virus particles could be recovered from nuclei primed with PEMV RNA. These results are discussed in relation to the possible mechanism for in vivo infection of pea cells.

Journal Article↗

HIV type 1 BF recombinant strains exhibit different pol gene mosaic patterns: descriptive analysis from 284 patients under treatment failure.

Different complex structures of the pol gene have been identified in 284 HIV-1 B/F recombinant sequences obtained from a group of 587 patients under treatment failure from Argentina. To analyze the mosaic structures of these viral sequences and to determine their phylogenetic relationship, the 284 partial pol gene sequences of BF recombinant viruses were amplified by RT-PCR and sequenced. Intersubtype breakpoints were analyzed by bootscanning. Phylogenetic relationships were determined by means of neighbor-joining trees. The analysis of the sequences showed multiple phylogenetic topologies clustering within intersubtype BF reference sequences. At least three different mosaic patterns were found compared to previously described BF-type viruses with unequal distribution in the studied population. The analysis also showed that HIV-1 BF recombinant viruses with diverse mosaic structures are phylogenetically related in their F segments and in selected B fragments with the F1 subtype and with BF recombinant viruses from Brazil, respectively, suggesting a common recombinant ancestor. No association was observed between the prevalence of each mosaic pattern and the frequency of major drug-resistance mutations in PR and RT.

Anti-HIV Agents↗

Mosaic analysis of the embryonic origin of taste buds.

The embryonic origins of taste receptor cells have not been established experimentally. Although related receptor cells (e.g. hair cells of the inner ear, lateral line receptors) are known to arise from neurogenic ectoderm (e.g. neural crest or placodes), taste buds are described as arising from local epithelial cells. Also unknown is whether or not each taste bud is a clone of cells, i.e. arising from a single progenitor. To address these problems, mosaic and chimeric analyses of lingual epithelium and taste buds have been undertaken. This paper describes the theory of chimeric and mosaic cell lineage analyses, the advantages and disadvantages, and the preliminary results obtained from the examination of the taste buds and lingual epithelium of: 1) mosaic Xenopus, 2) chimeric mice and 3) X-inactivation mosaic mice.

Animals↗

Stunting induced by cucumber mosaic cucumovirus-infected Nicotiana glutinosa is determined by a single amino acid residue in the coat protein.

R-CMV, a subgroup II strain of cucumber mosaic cucumovirus (CMV) induces a very strong stunting response in Nicotiana glutinosa plants, while Trk7-CMV causes green mosaic in this host. The genetic determinant of this phenotype was mapped to a 534-nucleotide region at the 3' end of RNA3 with biologically active, full-length cDNA clones of R-CMV and Trk7-CMV and RNA3 chimeras of the two strains. Within this region, R-CMV differs from Trk7-CMV by a single amino acid at position 193 in the coat protein. Changing the codon for Lys at this position to Asn or Ser, by site-directed mutagenesis, also changed the phenotype of the viruses from green mosaic to induction of stunting. Profound differences in both the spread and the accumulation of the viruses causing stunting and green mosaic were observed, although these did not correlate with the host specificity of stunting. Since expression of R-CMV coat protein with the PVX vector did not cause stunting, the data suggest that the presence of other CMV components is necessary for the induction of this symptom.

Amino Acid Sequence↗

Extended anterior thigh flaps for repair of massive cervical defects involving pharyngoesophagus and skin: an introduction to the "mosaic" flap principle.

"Mosaic" flaps, i.e., connected anteromedial thigh-groin flaps, and connected anterolateral thigh-medial thigh flaps, the pedicles of which are the perforators of the lateral circumflex femoral system, the superficial circumflex iliac vessels, and the perforators from the femoral vessels are used to repair cervical skin defects involving the pharyngoesophagus. Although they appear to be similar to "siamese" flaps, these mosaic flaps are a new concept because they consist of two adjacent flaps that are simultaneously elevated with double pedicles including the lateral circumflex femoral system. The pedicles of the groin or the medial thigh flaps are anastomosed to the muscle branch or the descending branch of the lateral circumflex femoral system, which is the pedicle of the anterior thigh flaps, and the system is joined to a single recipient vessel as a "bridge" flap. The main advantages of these mosaic flaps are (1) they offer single recipient vessels, (2) they offer a larger extension of the vascular territory of a skin flap, and (3) they offer the possibility of creating custom-made composite flaps such as musculocutaneous and osteocutaneous flaps, and (4) the elevation of the flaps simultaneous with head and neck tumor resection is possible. Based on cited cases, it is suggested that mosaic flaps are suitable to use for reconstruction of massive cervical defects involving the cervical skin and the pharyngoesophagus due to recurrent laryngeal carcinomas.

Aged↗

Mosaic expression of brush-border enzymes in infants with chronic diarrhea and malnutrition.

The chronic diarrhea observed in young malnourished infants that is sensitive to dietary glucose and other carbohydrates is associated with variable degrees of patchy mucosal villous atrophy. To explore intrinsic mucosal function in the pathogenesis of this alimentary intolerance, we have conducted an immunohistologic investigation of brush-border enzyme proteins of clinically obtained, mucosal biopsy samples. We used a group of monoclonal antibodies against human brush-border aminopeptidase, sucrase/isomaltase (SI), maltase, and lactase enzyme proteins. SI was strongly and uniformly expressed in crypts and villi of 11 of the 14 subjects; in 3 subjects, however, SI was expressed in a mosaic pattern. Maltase and lactase were occasionally absent, but more commonly were expressed in a mosaic distribution. The mosaic expression of brush-border enzyme proteins has been reported in congenital enzyme deficiencies associated with normal intestinal histology. We report the mosaic expression of brush-border enzyme proteins as a functional alteration associated with a pathological lesion of the mucosa in infants with chronic diarrhea. Our observation challenges the existing concept of ontogenic regulation of brush-border enzyme activity.

Aminopeptidases↗