Cell Wall Proteome of Sugarcane Young and Mature Leaves and Stems.

By characterizing the cell wall proteomes of different sugarcane organs (leaves and stems) at two developmental stages (young vs mature/apical vs basal), it is possible to address unique characteristics in each of them. Four-month-old leaves show a higher proportion of oxido-reductases and proteins related to lipid metabolism (LM), besides a lower proportion of proteins acting on polysaccharides, in comparison to 4-month-old internodes. It is possible to note that sugarcane leaves and young stems have the highest LM rate than all species, which is assumed to be linked to cuticle formation. The data generated enrich the number of cell wall proteins (CWPs) identified in sugarcane, reaching 277. To our knowledge, sugarcane has now the second higher coverage of monocot CWP in plants.

Characterising the Metabolomic Diversity and Biological Potentials of Extracts from Different Parts of Two Cistus Species Using UHPLC-MS/MS and In Vitro Techniques.

This study investigates the biochemical composition and biological properties of different parts (leaves, roots, and twigs) of two Cistus species (Cistus monspeliasis and Cistus parviflorus). The extracts were analysed using UHPLC-MS/MS to determine their chemical profiling. A range of antioxidant assays were performed to evaluate the extract's antioxidant capabilities. The enzyme inhibition studies focused on acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, and α-glucosidase and tyrosinase. In addition, the study examined the antimicrobial effects on different bacteria and yeasts and evaluated the toxicity using the MTT assay. Quinic acid, citric acid, gallic acid, catechin, quercetin derivatives, kaempferol, myricetin, ellagic acid, prodelphinidins, procyanidins, scopoletin, and flavogallonic acid dilactone are the main bioactive compounds found in both species. In enzyme inhibition assays, C. monspeliasis roots exhibited significant activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with the values of 2.58 ± 0.02 mg GALAE/g and 11.37 ± 1.93 mg GALAE/g, respectively. Cytotoxicity studies showed mostly weak toxicity, with some samples moderately reducing viability in RAW and HepG2 cells. These findings underscore the diverse biochemical profiles and bioactive potential of Cistus species, suggesting their utility as natural sources of antioxidants and enzyme inhibitors for pharmaceutical and nutraceutical development.

RhD variant caused by an in-frame triplet duplication in the RHD gene.

BACKGROUND: The RHD gene is highly polymorphic and a large number of D variants have already been detected. Several mechanisms are involved in the origin of D variants. In-frame deletions, resulting in a single-amino-acid deletion, have been described associated with RhD and RhCE variants. No in-frame duplications and/or insertions have been reported in the RH genes to date. STUDY DESIGN AND METHODS: Blood samples from a Brazilian blood donor and his sister were serologically tested with routine anti-D reagents and anti-D panels (ALBAclone advanced partial D typing kit, Alba Bioscience Limited; and D-Screen, Diagast), followed by molecular biology techniques, RHD polymerase chain reaction with sequence-specific priming and sequencing. RESULTS: Samples tested negative with routine immunoglobulin M (IgM) anti-D reagents and positive with IgG anti-D, which detect weak D cells. The pattern of results with anti-D panels did not correspond to any described before. A 3-bp in-frame duplication within Exon 1 (c.75_77dupTCT), resulting in the duplication of leucine 26 (p.Leu26dup), was identified in the two samples. CONCLUSION: We report the first RhD variant associated with a 3-bp in-frame duplication in the RHD gene, predicted to be located within the RhD protein transmembrane domain that might be expected to result in a weak-D-like phenotype, concordant with serologic findings.

Phase shifts, herbivory, and the resilience of coral reefs to climate change.

Many coral reefs worldwide have undergone phase shifts to alternate, degraded assemblages because of the combined effects of over-fishing, declining water quality, and the direct and indirect impacts of climate change. Here, we experimentally manipulated the density of large herbivorous fishes to test their influence on the resilience of coral assemblages in the aftermath of regional-scale bleaching in 1998, the largest coral mortality event recorded to date. The experiment was undertaken on the Great Barrier Reef, within a no-fishing reserve where coral abundances and diversity had been sharply reduced by bleaching. In control areas, where fishes were abundant, algal abundance remained low, whereas coral cover almost doubled (to 20%) over a 3 year period, primarily because of recruitment of species that had been locally extirpated by bleaching. In contrast, exclusion of large herbivorous fishes caused a dramatic explosion of macroalgae, which suppressed the fecundity, recruitment, and survival of corals. Consequently, management of fish stocks is a key component in preventing phase shifts and managing reef resilience. Importantly, local stewardship of fishing effort is a tractable goal for conservation of reefs, and this local action can also provide some insurance against larger-scale disturbances such as mass bleaching, which are impractical to manage directly.

New species of Stylopoma (Bryozoa, Cheilostomata) from Bahia State, northeastern Brazil.

At date, seven species of Stylopoma Levinsen, 1909 are reported from the Brazilian coast, five of which are known from Bahia State in the northeastern region. Here we describe four new species of Stylopoma, all from Bahia State: Stylopoma corallinum n. sp., Stylopoma faceluciae n. sp., Stylopoma multiavicularia n. sp. and Stylopoma sinuata n. sp. Unlike the Caribbean species of Stylopoma, that mainly have rounded primary orifice and slit-like sinus, the morphology of the primary orifice of Brazilian taxa, including the new species presented here, is quite variable. In the newly described taxa, the sinus is drop-shaped in S. corallinum n. sp. and S. faceluciae n. sp., U-shaped in S. multiavicularia n. sp. and V-shaped in S. sinuata n. sp. Like congeners from the Caribe, S. corallinum n. sp., S. faceluciae n. sp. and S. multiavicularia n. sp. have avicularia on the surface of the ooecia. Remarks on the morphological characters currently used in Stylopoma taxonomy are provided, including a comparative table of all living species worldwide.

Faecal Indicator Bacteria and Pseudomonas aeruginosa in Marine Coastal Waters: Is there a Relationship?

To estimate the quality of coastal waters, European Union Directive 2006/7/EC provides guidelines to assess levels of faecal bacteria, including Escherichia coli and intestinal enterococci. These microbiological criteria are based on studies that determine the risk of bathers having diseases caused by enteric bacteria, not necessarily measuring the potential danger associated with the presence of nonenteric pathogens. The association between the presence of faecal contaminant indicators and nonenteric pathogenic microorganisms has not been well defined yet. The purpose of this study is to establish a relationship between Pseudomonas aeruginosa and microbiological indicators of faecal contamination. Presence of microbiological contamination in the coastal waters near the sewage treatment plant (STP) of Peniche (Portugal) was confirmed (P. aeruginosa 135.8 Colony Forming Unit/100 mL, Escherichia coli 1100.1 Most Probable Number/100 mL, intestinal enterococci 2685.9 MPN/100 mL) with much lower levels in the areas located south of the STP, along the main water coastal current (beach 1: 0.7 CFU/100 mL, 16.5 MPN/100 mL, 100.5 MPN/100 mL; beach 2: 0.3 CFU/100 mL, 74.0 MPN/100 mL, 145.9 MPN/100 mL, respectively). Analysis of Pearson's correlation revealed a strong positive correlation between E. coli and P. aeruginosa, suggesting E. coli as an indicator of P. aeruginosa presence.

Antitubercular and anti-inflammatory properties screening of natural products from Plectranthus species.

AIM: Confirm the use of Plectanthus spp. plants in traditional medicine, particularly as anti-inflammatory and anti-infective agents. MATERIALS & METHODS: Compounds previously isolated from Plectranthus spp. were studied for their anti-inflammatory activity using the SNAP assay and RAW 264.7 cells, by the quantification of nitric oxide. An halimane diterpene and its derivatives were tested in infected macrophages with M. tuberculosis H37Rv, using CFU counts assay, at their minimum inhibitory concentration values. Results: The isolated compounds tested at noncytotoxic concentrations, did not reveal nitric oxide scavenging in the S-nitroso-N-acetylpenicillamine and the cellular assays. On the other hand, promising results were obtained regarding one semisynthetic halimane derivative (11R*,13E)-halima-5,13-diene-11,15-diol), previously prepared (2.1 × 105 CFU/mL), with an effect similar to the antitubercular drugs ethambutol (2.0 × 105 CFU/mL) and isoniazid (1.2 × 105 CFU/mL). CONCLUSION: The present report demonstrates the relevance of Plectranthus spp. in medicinal chemistry drug development for TB and other infective respiratory complaints. Also, this work suggests that further studies involving other inflammatory mediators are needed to validate the anti-inflammatory use of these medicinal plants.

ABO exon and intron analysis in individuals with the AweakB phenotype reveals a novel O1v-A2 hybrid allele that causes four missense mutations in the A transferase.

BACKGROUND: Since the cloning in 1990 of cDNA corresponding to mRNA transcribed at the blood-group ABO locus, polymorphisms due to ethnic and/or phenotypic variations have been reported. Some subgroups have been explained at the molecular level, but unresolved samples are frequently encountered in the reference laboratory. RESULTS: ABO blood grouping discrepancies were investigated serologically and by ABO genotyping [duplex polymerase-chain-reaction (PCR)--restriction-fragment-length-polymorphism (RFLP) and PCR--allele-specific-primer (ASP) across intron 6] and DNA sequencing of the ABO gene and its proposed regulatory elements. Blood samples from five individuals living in Portugal, Switzerland, Sweden and the USA were analysed. These individuals were confirmed to be of Black ethnic origin and had the unusual AweakB phenotype but appeared to have the A2B genotype without previously reported mutations associated with weak A or B expression. Sequencing of this A allele (having 467C>T and 1061delC associated with the common A2 [A201] allele) revealed three mutations regularly encountered in the O1v [O02] allele: 106C>T (Val36Phe), 188G>A (Arg63His), 220C>T (Pro74Ser) in exons 3, 4 and 5, respectively. The additional presence of 46G>A (Ala16Thr) was noted, whilst 189C>T that normally accompanies 188G>A in O1v was missing, as were all O1v-related mutations in exons 6 and 7 (261delG, 297A>G, 646T>A, 681G>A, 771C>T and 829G>A). On screening other samples, 46G>A was absent, but two new O alleles were found, a Jordanian O1 and an African O1v allele having 188G>A but lacking 189C>T. Sequencing of introns 2, 3, 4 and 5 in common alleles (A1 [A101], A2, B [B101], O1, O1vand O2 [O03]) revealed 7, 12, 17 and 8 polymorphic positions, respectively, suggesting that alleles could be defined by intronic sequences. These polymorphic sites allowed definition of a breakpoint in intron 5 where the O1v-related sequence was fused with A2 to form the new hybrid. Intron 6 has previously been sequenced. Four new mutations were detected in the hybrid allele and these were subsequently also found in intron 6 of A2 alleles in other Black African samples. CONCLUSIONS: A novel O1v-A2 hybrid was defined by ABO exon/intron analysis in five unrelated individuals of African descent with the AweakB blood group phenotype.

Standardizing clinical trials workflow representation in UML for international site comparison.

BACKGROUND: With the globalization of clinical trials, a growing emphasis has been placed on the standardization of the workflow in order to ensure the reproducibility and reliability of the overall trial. Despite the importance of workflow evaluation, to our knowledge no previous studies have attempted to adapt existing modeling languages to standardize the representation of clinical trials. Unified Modeling Language (UML) is a computational language that can be used to model operational workflow, and a UML profile can be developed to standardize UML models within a given domain. This paper's objective is to develop a UML profile to extend the UML Activity Diagram schema into the clinical trials domain, defining a standard representation for clinical trial workflow diagrams in UML. METHODS: Two Brazilian clinical trial sites in rheumatology and oncology were examined to model their workflow and collect time-motion data. UML modeling was conducted in Eclipse, and a UML profile was developed to incorporate information used in discrete event simulation software. RESULTS: Ethnographic observation revealed bottlenecks in workflow: these included tasks requiring full commitment of CRCs, transferring notes from paper to computers, deviations from standard operating procedures, and conflicts between different IT systems. Time-motion analysis revealed that nurses' activities took up the most time in the workflow and contained a high frequency of shorter duration activities. Administrative assistants performed more activities near the beginning and end of the workflow. Overall, clinical trial tasks had a greater frequency than clinic routines or other general activities. CONCLUSIONS: This paper describes a method for modeling clinical trial workflow in UML and standardizing these workflow diagrams through a UML profile. In the increasingly global environment of clinical trials, the standardization of workflow modeling is a necessary precursor to conducting a comparative analysis of international clinical trials workflows.

Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA.

UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis.

Qualidade de vida e sobrecarga de cuidadores de crianças com câncer / Quality of life and work overload of caretakers of children with cancer / Calidad de vida y sobrecarga de cuidadores de niños con câncer

O objetivo do estudo foi verificar a relação entre a qualidade de vida e a sobrecarga em cuidadores de crianças portadoras de neoplasia e assistidas em uma casa de apoio. A metodologia foi quantiqualitativa com a utilização de multimétodos de pesquisa. Os participantes (N = 23) foram os cuidadores dos pacientes acompanhados pela Casa de Apoio à Criança com Câncer. Os instrumentos de coleta de dados foram: um questionário sociodemográfico, a Escala Whoqol - Bref, a Escala Zarit e uma entrevista por pautas. A análise dos dados se deu pela estatística descritiva e pela análise da enunciação. Os resultados apontam uma amostra essencialmente feminina (95,7%); sendo 82,6% formada pelas mães. A maioria apresentou sobrecarga intensa (54,5%). O domínio de maior prejuízo foi o psicológico (53,98). As entrevistas confirmaram os dados obtidos nas escalas e mostraram mudanças significativas na vida dos cuidadores e da família, com prejuízo marcadamente para a vida social e a relação de trabalho. Os cuidadores sentem-se sobrecarregados, mas a sobrecarga permanece velada, já que a sua manifestação pode revelar emoções e sentimentos de culpa, difíceis de serem admitidos e elaborados.

This paper aims at verifying the relationship between the quality of life and work overload of caretakers of children with neoplasia that are cared for at a supporting home. The methodology was quantity-qualitative with use of multi methods of research. The participants (N = 23) were the caretakers of patients cared for at the Support Home for Children with Cancer. The data collection instruments were: a socio-demographic questionnaire, the Whoqol - Bref Scale, the Zarit Scale, and one interview by topics. Data analysis was done by the descriptive statistics and by the enunciation analysis. The outcomes point to an essentially female sample (95.7%); while mothers comprised 82.6%. The majority presented intense overload (54.5%). The predominant loss was psychological (53.98). The inerviews confirmed that obtained in the scaales and they showed significant changes in the life of caretakers and families, with marked losses for social life and work relations. The caretakers feel overloaded, but this overload remains disguised, as its manifestation may reveal emotions and guilty feelings, which are hard to be admitted and elaborated.

El objetivo del estudio fue a verificar la relación entre la calidad de vida y la sobrecarga en cuidadores de niños portadores de neoplasia y asistidas en una casa de apoyo. La metodología fue cuanti-cualitativa con la utilización de multimétodos de pesquisa. Los participantes (N = 23) fueron los cuidadores de los pacientes acompañados por la Casa de Apoyo a Niños con Cáncer. Los instrumentos de colecta de datos fueron: un cuestionario socio demográfico, la Escala Whoqol - Bref, la Escala Zarit y una entrevista por pautas. El análisis de los datos se dio por la estadística descriptiva y por el análisis de la enunciación. Los resultados apuntan una muestra esencialmente femenina (95,7%); siendo un 82,6% formada por las madres. La mayoría presentó sobrecarga intensa (54,5%). El dominio de mayor perjuicio fue el psicológico (53,98). Las entrevistas confirmaron los datos obtenidos en las escalas y mostraron mudanzas significativas en la vida de los cuidadores y de la familia, con perjuicio marcadamente para la vida social y la relación de trabajo. Los cuidadores se sienten sobrecargados, pero la sobrecarga permanece velada, ya que su manifestación puede revelar emociones y sentimientos de culpa, difíciles de ser admitidos y elaborados.

Identification of six new alleles at the FUT1 and FUT2 loci in ethnically diverse individuals with Bombay and Para-Bombay phenotypes.

BACKGROUND: The Bombay and para-Bombay phenotypes arise from mutations of the FUT1 gene that silence the gene or affect the efficiency of the encoded 2-alpha-fucosyltransferase. Samples from seven individuals of different geographic backgrounds whose red blood cells had an apparent Bombay or para-Bombay phenotype were investigated. Among these, novel FUT1 and FUT2 alleles were identified. STUDY DESIGN AND METHODS: Standard serologic techniques were used. Genomic DNA was sequenced with primers that amplified the coding sequence of FUT1 and the related secretor gene, FUT2. Routine ABO genotyping analysis was performed. RESULTS: Five new FUT1 alleles were identified that silenced FUT1 or weakened alpha2FucT1 activity. These were 35C>T, 269G>T (Ala11Val, Gly89Val); 421A>G (Trp140Stop); 538C>T, 1089T>G (Gln180Stop, Ala363Ala); 689A>C (Gln230Pro); and 917C>T (Thr305Ile). In addition, both homozygosity and heterozygosity for the previously reported mutation, 826C>T (Gln276Stop), were observed. Four of seven samples were homozygous for the silencing mutation 428A in FUT2. One new FUT2 allele was identified: 278C>T, 357C>T (Ala93Val, Asn119Asn). CONCLUSIONS: These results add to the growing database of apparently sporadic and random mutations in the FUT1 gene and confirm previous reports regarding the lack of ethnic bias. In contrast, our data reinforce the apparent maintenance of the common nonsecretor FUT2 alleles in the population.

Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA

UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis.