[Morbid obesity and surgery: state of the art].
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Biliopancreatic Diversion↗
Biomedical subjects
P Moreno
Publications and source records attributed to P Moreno.
Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes.
Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recovery of transgenic plants showing gene silencing in woody species, Mexican lime [ Citrus aurantifolia (Christm.) Swing.] plants were transformed with the p25 coat protein gene of Citrus tristeza virus (CTV) with or without selection for nptII and uidA. Strikingly, more than 30% of the transgenic limes regenerated under non-selective conditions had silenced transgenes, and in all cases silencing affected all the three transgenes incorporated. These results indicate that the frequency of transgene silencing may be greatly underestimated when the rate of silencing is estimated from the number of regenerants obtained under selective conditions. To our knowledge, this is the first report in which the frequency of gene silencing after transformation has been quantified. When the integration pattern of T-DNA was analyzed in silenced and non-silenced lines, it was observed that inverted repeats as well as direct repeats and even single integrations were able to trigger gene silencing. Gene silencing has often been associated with the insertion of DNA sequences as inverted repeats. Interestingly, here, direct repeats and single-copy insertions were found in both silenced and non-silenced lines, suggesting that the presence of inverted-repeat T-DNAs and the subsequent formation of dsRNAs triggering gene silencing cannot account for all silencing events.
Citrus↗
Behaviour of pesticides in a water/sediment system under laboratory and field conditions.
Experiments were conducted in order to evaluate the behaviour of some pesticides commonly used in rice production. At a laboratory scale, but exposed to the environmental conditions, a known concentration of pesticides in water flowed through a cement channel over a sediment layer, at a constant flow rate. The collected water was homogenised and analysed by gas chromatography/HPLC to assess the concentration of each pesticide. Under these conditions, the reduction of each pestcide concentration in water depends on the nature of it and on the water flow rate. The lower the flow rate, the grater the reduction in the concentration of pesticides in the water that ran over the sediment layer. Experiments were also carried out under field conditions, using a 1200 m length, 2 m width and 0.3 m depth rice field channel. A pesticide mixture of known concentration was poured on the flowing water and mixed thoroughly. After the arrival of this contaminated water at preestablished sampling points down the stream, the concentration of each pesticide in water was determined. The longer the distance of the sampling points downstream from the starting point, the lower the pesticide concentration found. Only a few pesticides were detected at the 1200 m sampling point, but at very low concentration. The others were not detected. It was pointed out that the channel length was the most important factor in the reduction of the pesticide concentration in the flowing water. The influence of the flow rate on this reduction could not be clearly related as in the laboratory experiments.
Agriculture↗
Diversity and biomass of chironomidae (Diptera) larvae in an impacted coastal lagoon in Rio De Janeiro, Brazil.
Diversity and biomass of Chironomidae larvae were studied between January-November 1993 and March-November 1994 in an impacted lagoon in Rio de Janeiro, Southeastern Brazil, in an attempt to establish the ecological consequences of anthropogenic eutrophication processes. Nine hundred and seventy-five organisms belonging to the Tanypodinae, Orthocladiinae, and Chironominae were collected. Polypedilum (62%) and Chironomus (58%) were the most common genera found in the limnetic and littoral zones.
Animals↗
DYS19, DYS385, DYS389 (I and II), DYS390, DYS391, DYS392, DYS393 haplotypes in a population sample from Catalonia (north-east Spain).
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Forensic Medicine↗
Reductive fragmentation of carbohydrate anomeric alkoxy radicals. Synthesis of alditols with potential utility as chiral synthons.
A series of anomeric nitrate esters and N-phthalimido glycosides of carbohydrates in furanose and pyranose forms have been synthesized in order to generate the corresponding alkoxy radicals and study the C1-C2 fragmentation reaction under reductive conditions. This reaction constitutes a two-step method for the transformation of carbohydrates into the corresponding alditols with one less carbon. Using this methodology, interesting four- and five-carbon building blocks for natural products synthesis possessing D-erythritol, D-threitol, D-xylitol, and D-arabinitol stereochemistry have been prepared. The synthesis of 1,2-O-isopropylidene-beta-L-threose (40) and 1-acetamido-2,4,5-tri-O-acetyl-D-arabinitol (50) have also been achieved from 1,2:5,6-di-O-isopropylidene-beta-D-glucofuranose and 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-D-glucopyranose, respectively.
Alcohols↗
The nucleotide sequence and genomic organization of Citrus leaf blotch virus: candidate type species for a new virus genus.
The complete nucleotide sequence of Citrus leaf blotch virus (CLBV) was determined. CLBV genomic RNA (gRNA) has 8747 nt, excluding the 3'-terminal poly(A) tail, and contains three open reading frames (ORFs) and untranslated regions (UTR) of 73 and 541 nucleotides at the 5' and 3' termini, respectively. ORF1 potentially encodes a 227.4-kDa polypeptide, which has methyltransferase, papain-like protease, helicase, and RNA-dependent RNA polymerase motifs. ORF2 encodes a 40.2-kDa polypeptide containing a motif characteristic of cell-to-cell movement proteins. The 40.7-kDa polypeptide encoded by ORF3 was identified as the coat protein. The genome organization of CLBV resembles that of viruses in the genus Trichovirus, but they differ in various aspects: (i) in trichoviruses ORF2 overlaps ORFs 1 and 3, whereas in CLBV, ORFs 2 and 3 are separated and ORFs 1 and 2 overlap in one nucleotide; (ii) CLBV gRNA and CP are larger than those of trichoviruses; and (iii) the CLBV 3' UTR is larger than that of trichoviruses. Phylogenetic comparisons based on CP amino acid signatures clearly separates CLBV from trichoviruses. Also contrasting with trichoviruses, CLBV could not be transmitted to Chenopodium quinoa Willd. Considering these singularities, we propose that CLBV should be included in a new virus genus.
Amino Acid Sequence↗
Amplification of Citrus tristeza virus from a cDNA clone and infection of citrus trees.
Isolates of the Closterovirus, Citrus tristeza virus (CTV), are populations of disparate genotypes and defective RNAs developed during long periods of vegetative propagation of citrus trees. Because it has not been possible to obtain pure cultures of the virus, it is not known what components of the population are primarily responsible for induction of diseases. We previously developed an infectious cDNA clone from which in vitro-produced RNA transcripts could infect protoplasts (Satyanarayana et al., 1999, Proc. Natl. Acad. Sci. USA 96, 7433-7438). However, neither the RNA transcripts nor virions from transcript-infected protoplasts were competent for infection of citrus trees. Using a green fluorescent protein-marked virus as inoculum, we found that the approximately 20-kb RNA from virions or transcripts of cDNA infected only a small percentage of protoplasts ( approximately 0.01%), but virions could infect more than 80% of the protoplasts. Based on this information, we amplified the virus from the cDNA clone (recombinant virus) by successive passages in protoplasts using virions in crude sap as inoculum. By the third to seventh passages in protoplasts, maximal amounts of recombinant progeny virus were produced, which were used for inoculation of small citrus trees by slashing stems in the presence of virion preparations. A relatively high percentage of plants became infected with the recombinant virus from protoplasts, resulting in the first defined pure culture of CTV in plants. The comparative biology of the pure culture of recombinant CTV with that of the parental population in planta demonstrated that the recombinant virus retained through all of the recombinant DNA manipulations the normal functions of replication, movement, and aphid transmissibility, and had a symptom phenotype indistinguishable from that of the parental population. Additionally, fulfilling Koch's postulates of the first pure culture of CTV in plants suggested that the major genotype of the CTV T36 population is the primary determinant of the symptom phenotype. We could distinguish no biological contributions resulting from the minor genotypes and defective RNAs of the parental population.
Animals↗
The Bubi population of Equatorial Guinea characterised by HUMTH01, HUMVWA31A, HUMCSF1PO, HUMTPOX, D3S1358, D8S1179, D18S51 and D19S253 STR polymorphisms.
Allele frequencies for eight STR loci (HUMTH01, HUMVWA31A, HUMCSF1PO, HUMTPOX, D3S1358, D8S1179, D18S51, D19S253) have been analysed in the Bubi population of Bioko Island, Equatorial Guinea. For all loci, no deviation from Hardy-Weinberg equilibrium was found. Data obtained were compared with that of Caucasian and African populations. Significant differences were found for all systems between all the black populations compared and the Caucasoid population. Similarities were observed between the Bubi and Zimbabweans, and also with African American populations. Also, more affinities were observed between Zimbabweans and Ugandans and Ovambos than between these groups and the Bubi population. From these comparisons it is suggested that in Africa, as in other continents, there is a certain genetic heterogeneity.
Black People↗
Partial characterisation of citrus leaf blotch virus, a new virus from Nagami kumquat.
Citrus leaf blotch virus (CLBV) was purified from leaves of Nagami kumquat SRA-153 that showed bud union crease when propagated on Troyer citrange. Virions were filamentous particles (960 x 14 nm) containing a 42 kDa protein and a single-stranded RNA (ssRNA) of about 9,000 nt (Mr 3 x 10(6)). Infected tissue contained three species of double-stranded RNA (dsRNA) of Mr 6, 4.5 and 3.4 x 10(6). The nucleotide sequence of several complementary DNA (cDNA) clones showed significant similarities with replication-related proteins from plant filamentous viruses in several genera. A digoxigenin-labelled probe from one of these cDNA clones hybridised in Northern blots with ssRNA from virions and with the three dsRNA species, suggesting that the ssRNA is the genomic RNA of the virus, the largest dsRNA is its replicative form, and the two smaller dsRNAs probably replicative forms of 5' co-terminal subgenomic RNAs. CLBV was also detected in several citrus cultivars from Spain and Japan including Navelina sweet orange field trees propagated on Troyer citrange showing bud union crease; however, no virus could be detected in other citrus trees with similar symptoms. This indicates that CLBV is not restricted to kumquat SRA-153, but its involvement in causing the bud union disorder remains unclear.
Citrus↗
Polymorphism of the 5' terminal region of Citrus tristeza virus (CTV) RNA: incidence of three sequence types in isolates of different origin and pathogenicity.
Sequences of the 5' terminal region of the genomic RNA from eight isolates of Citrus tristeza virus (CTV) were previously classified into three types (I, II and III), with intragroup sequence identity higher than 88% and intergroup sequence identity as low as 44%. Sequencing of an additional 58 cDNA clones from 15 virus isolates showed that all sequences could be unequivocally assigned to one of the three types previously established. The relative frequency of each sequence type was assessed in 57 CTV isolates of different geographic origin and pathogenic characteristics by RT-PCR with sets of type-specific primers using CTV dsRNA as template. None of the isolates yielded amplification of the type I or II sequences alone, but in 19 of them type III sequences were the only amplification product detected. Within isolates containing more than one sequence type, eight had type II and III sequences, 11 had type I and III sequences, and 19 had sequences of the three types. Isolates containing only type III sequences caused only mild to moderate symptoms in Mexican lime, an indicator species for most CTV isolates, whereas isolates causing stem pitting in sweet orange an/or grapefruit, generally contained sequences type II. None of the sequence types could be traced to a precise geographic area, as all types were detected in isolates from at least nine of the 12 countries from which samples were taken.
5' Untranslated Regions↗
Genetic variation of Citrus tristeza virus isolates from California and Spain: evidence for mixed infections and recombination.
We examined the population structure and genetic variation of four genomic regions within and between 30 Citrus tristeza virus (CTV) isolates from Spain and California. Our analyses showed that most isolates contained a population of sequence variants, with one being predominant. Four isolates showed two major sequence variants in some genomic regions. The two major variants of three of these isolates showed very low nucleotide identity to each other but were very similar to those of other isolates, suggesting the possibility of mixed infections with two divergent isolates. Incongruencies of phylogenetic relationships in the different genomic regions and statistical analyses suggested that the genomes of some CTV sequence variants originated by recombination events between diverged sequence variants. No correlation was observed between geographic origin and nucleotide distance, and thus from a genetic view, the Spanish and Californian isolates analyzed here could be considered members of the same population.
California↗
Habitat diversity and benthic functional trophic groups at Serra do Cipó, Southeast Brazil.
The assessment of the diversity of habitats and the characterisation of the functional trophic groups of benthic macroinvertebrate communities of some rivers of Serra do Cipó (MG) were the main objectives of this study. The available trophic resources and the types of substrata were characterised along with the structure and composition of their using functional trophic groups. Serra do Cipó is a watershed divisor of the São Francisco and Doce River basins, including a series of streams and rivers, of good water quality and well preserved ecological characteristics. Samples were collected in Cipó, Peixe and Preto do Itambé rivers, besides the Indaiá and Capão da Mata streams at 26 sampling stations, during the rainy (February) and dry (October) seasons of 1998, using "Kicking nets" of 0.125 mm mesh size. The group of collectors (Baetidae, Leptophlebiidae and Leptohyphidae) was the most abundant, followed by collector-predators (Hydrophilidae, Ceratopogonidae, Chironomidae-Tanypodinae), and detritivorous-herbivores (Oligochaeta). The riparian vegetation, together with the aquatic macrophytes, are the substrata containing the highest richness of functional trophic groups and the higher habitat diversity. The results suggest that the use of functional trophic groups, together with habitat evaluation, are efficient tools in the evaluation of the diversity of benthic macroinvertebrates, particularly in altitudinal lotic ecosystems.
Animals↗
Allele frequency for three STR loci in a population sample from Catalonia (Spain) using a simple manual triplex PCR method.
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DNA Fingerprinting↗
Results of the management of upper gastrointestinal bleeding from gastroesophageal varices.
INTRODUCTION: The management of upper gastrointestinal bleeding caused by rupture of gastric and/or esophageal varices in patients with liver cirrhosis must focus on the initial control of the haemorrhage avoiding further worsening of an already poor liver function and the prevention of early relapsing bleeding. Therapeutic options include endoscopic, pharmacological and surgical methods. MATERIAL AND METHODS: Prospective study of the results obtained after the follow-up of 90 bleeding episodes in a total of 54 patients, 35 men and 19 women, with a mean age of 58 years (range 32-77), to which a therapeutic protocol for acute bleeding secondary to portal hypertension was applied over a 22-months period. Patient classification according to Child-Pugh upon admission was 57% Child A, 34% Child B and 9% Child C. RESULTS: Mean hospital length of stay was 9 days (2-50). Of the 90 bleeding episodes, 15 were early relapsing bleeding episodes (16.7%). Twelve patients died (mortality rate of 22.2% by patients and 13.4% by bleeding episodes). Twelve emergency surgical procedures were performed because of the persistence of haemorrhage. Forty one per cent of patients were readmitted because of relapsing bleeding at least once during the follow-up period. CONCLUSIONS: Management of upper gastrointestinal bleeding due to gastroesophageal varices in patients with liver cirrhosis requires a combined therapy in order to attain maximum effectiveness in acute haemorrhagic episodes and to address all potential later consequences. Such therapy should be provided in a hospital fully equipped and with specialists in this pathology. Based on our experience, emergency surgery as rescue treatment for persistent or short-term relapsing bleeding should be restricted to patients with good hepatic function because of its high morbidity and mortality.
Adult↗
The p20 gene product of Citrus tristeza virus accumulates in the amorphous inclusion bodies.
Citrus tristeza virus (CTV) has 10 3' open reading frames (ORFs) of unknown function except for the two coat proteins. The highest produced subgenomic RNAs are those of the major coat protein gene (p25) and the 3' most genes, p20 and p23. The proteins from three ORFs, p25, p27, and p20, were examined in the yeast two-hybrid assay for the interactions between themselves and to one another. The p20 protein exhibited a high affinity for itself, suggesting that it might aggregate in infected cells. The cytopathology of CTV infections includes characteristic paracrystalline and amorphous inclusions in the phloem elements of infected citrus. Polyclonal antiserum raised against the bacterial expressed p20 gene product detected a protein of approximately 22-23 kDa, which accumulated to relatively high levels in CTV-infected citrus, but not in healthy citrus. Immunogold localization using antibodies to p20 protein showed strong and specific labeling of the amorphous inclusion bodies present in CTV-infected cells. Mesophyll protoplasts of Nicotiana benthamiana transfected with a CTV mutant containing the green fluorescent protein (GFP) ORF fused in-frame to the 3' end of p20 protein ORF expressed high levels of GFP. The fusion protein was concentrated in one specific area in the cytoplasm and lacked an organized shape. Accumulation of high levels of p20 protein in infected tissue, specific localization of the p20-GFP fusion protein, immunolocalization of p20 protein into amorphous inclusions, and strong homologous p20 protein-p20 protein interactions in the yeast-two-hybrid assay suggest that the p20 protein of CTV is a major component of the amorphous inclusion bodies present in CTV-infected cells.
Blotting, Western↗
The 23-kDa protein coded by the 3'-terminal gene of citrus tristeza virus is an RNA-binding protein.
The 23-kDa protein (p23), encoded by the 3'-proximal gene of the RNA of Citrus tristeza virus (CTV), was overexpressed in Escherichia coli fused to the maltose-binding protein and purified by affinity chromatography. Gel retardation and UV crosslinking assays demonstrated that p23 has the ability to cooperatively bind single-stranded RNA in a non-sequence-specific manner. Formation of the p23-RNA complex was dependent on the conformational state of p23 and on the presence of a basic region, but the complex was stable at high salt concentrations, suggesting that interactions other than those between the negatively charged RNA and the basic region of p23 are involved. Competition assays showed that the affinity of p23 for single-stranded and double-stranded RNA was similar but considerably higher than for single-stranded and double-stranded DNA. By use of a series of artificially generated mutants, the RNA-binding domain of p23 was mapped between positions 50-86, a region containing several basic amino acids and a putative zinc-finger domain. Additional p23-derivatives lacking the conserved residues presumably involved in coordinating the zinc ion showed RNA-binding activity, but with an apparent dissociation constant higher than the wild-type protein. These conserved residues might confer binding specificity or increase binding stability in vivo. Within the Closteroviridae family, p23 is the only protein characterized so far showing RNA-binding activity.
Chromosome Mapping↗
[Estimate of HIV-1 infection prevalence in pregnant women and effectiveness of zidovudine administered during pregnancy in the prevention of vertical transmission].
BACKGROUND: Despite the proven efficacy of zidovudine (ZDV) for reducing perinatal transmission of HIV-1, questions remain about its implementation and effectiveness in routine practice. The aims of this study were to assess the impact of ZDV administered during pregnancy in preventing perinatal HIV-1 transmission, and to determine the proportion of early identification of maternal HIV-1 infection over time. PATIENTS AND METHODS: We prospectively followed from birth a cohort of children born between 1/1/1987 and 31/10/1997 to HIV-1-infected mothers. Infant infection status was assessed by follow-up beyond 18 months or HIV-PCR up to 3 months of age. RESULTS: 229 mothers and 248 infants were identified in the cohort. ZDV was administered during pregnancy as monotherapy to 34 mothers for a mean of 4.7 (3.1) months prior to delivery. There were no differences in baseline characteristics between the treated and untreated groups. Mean (SD) CD4 cell count was 465 (261) cells x 10(6)/l. Factors associated with transmission were a more prolonged time of rupture of obstetric membranes (median 6 vs 1.04 hours; p = 0.023) and ZDV treatment. Among the ZDV-treated mothers only one child was infected (2.9%), whereas 37 children born to 212 untreated women became infected. (OR: 0.14; 95% CI: 0.07-0.92). The estimated prevalence of HIV-1 in pregnant women in our area is about 0.39% (95% CI: 0.34-0.45). From 1987 to 1991, 9.7% of infected women were nor identified at the perinatal period, as compared to 2.5% in the last 5 years of the study period (p = 0.034). CONCLUSIONS: In this study, the estimated prevalence of HIV-1 infection is high. ZDV during pregnancy is significantly associated with a decrease in perinatal transmission in our setting. The awareness of an effective treatment might have contributed to the increased identification of HIV-infected mothers prior to delivery observed over time in our cohort of children born to HIV-infected mothers.
Adolescent↗