Study of defense-related gene expression in grapevine infested by <Emphasis Type="Italic">Colomerus vitis</Emphasis> (Acari: Eriophyidae)

As the main source of lipids and proteins in honey bees, pollen is a major nutrient provider involved in development and health and has been studied for tolerance stimulation against pathogens and parasites. In the case of Varroa destructor Anderson & Trueman (Acari, Mesostigmata: Varroidae) parasitization, the lack of a complete laboratory system to rear both the bee larva and the acarian parasite limited the studies concerning larval nutrition effects on the bee tolerance and resistance against varroatosis. Due to the development of this complete rearing protocol, we managed to feed young honey bee larvae with pollen supplemented solutions and to study the effect on their later development under parasitism conditions. In our experimental conditions, pollen influences neither the deformity rate, nor the survival of bees both parasitized and unparasitized. However, pollen extract supplementation seems to significantly impact the weight of the spinning bee larvae without having an effect on the physiological weight loss during pupation, so the differences found at the larval stage remain the same as at emergence. Varroa has a deleterious effect on bee pupae and led to a steady increase of the physiological weight loss experienced during metamorphosis. Interestingly, this ponderal loss associated with Varroa parasitization seems to be reduced in the polyfloral pollen supplementation condition. Altogether, this work is to our knowledge the first to study in laboratory conditions the impact of larval nutrition on the tolerance to parasitism. A diverse pollen diet may be beneficial to the bees’ tolerance against V. destructor parasitism.     

Total phenolics,resveratrol content and antioxidant activity of seeds and calluses of pinto peanut (<Emphasis Type="Italic">Arachis pintoi</Emphasis> Krapov. &amp; W.C. Greg.)

Arachis pintoi is a peanut species native to Brazil, which is cultivated in many countries for animal forage, soil cover, landscaping, and recovery of degraded areas. Tissue culture studies for this species have been focused in plant production, whereas works on in vitro secondary metabolites production are scarce. The goal of the present work was to establish callus cultures from different seed explants of A. pintoi, aiming at evaluating the potential for metabolites production and antioxidant activity. Embryonic axes, leaflets, and cotyledons were cultured on solidified MS medium supplemented with picloram (PIC), 2,4-dichlorophenoxyacetic acid (2,4-D), thidiazuron (TDZ) or different combinations of 6-benzyladenine (BA) and α-naphthaleneacetic acid (NAA), under light or dark conditions. Friable calluses with a high biomass (4.3?±?0.3 g FW per callus) were obtained from embryonic leaflets cultured on medium supplemented with 17.6 µM BA plus 5.4 µM NAA, in the dark. Cotyledons and embryonic axes cultured in the presence of 4.4 µM BA combined with 10.8 µM NAA formed heterogeneous calluses with a compact base and a large friable surface. Trans-resveratrol and other stilbenes that were not found in seeds were detected in callus extracts, especially those originated from cotyledons, although these materials showed lower total phenolic contents (TPC) when compared with seeds with and without testa, as well as cotyledons. Extracts from seeds with testa and from calluses derived from cotyledons and embryonic axes showed the highest EC₅₀ in DPPH assays. No correlation between TPC, trans-resveratrol and antioxidant activity was observed.     

Comparative proteomic analysis of Xanthomonas citri ssp. citri periplasmic proteins reveals changes in cellular envelope metabolism during in vitro pathogenicity induction

Citrus canker is a plant disease caused by Gram‐negative bacteria from the genus Xanthomonas. The most virulent species is Xanthomonas citri ssp. citri (XAC), which attacks a wide range of citrus hosts. Differential proteomic analysis of the periplasm‐enriched fraction was performed for XAC cells grown in pathogenicity‐inducing (XAM‐M) and pathogenicity‐non‐inducing (nutrient broth) media using two‐dimensional electrophoresis combined with liquid chromatography‐tandem mass spectrometry. Amongst the 40 proteins identified, transglycosylase was detected in a highly abundant spot in XAC cells grown under inducing condition. Additional up‐regulated proteins related to cellular envelope metabolism included glucose‐1‐phosphate thymidylyltransferase, dTDP‐4‐dehydrorhamnose‐3,5‐epimerase and peptidyl‐prolyl cis–trans‐isomerase. Phosphoglucomutase and superoxide dismutase proteins, known to be involved in pathogenicity in other Xanthomonas species or organisms, were also detected. Western blot and quantitative real‐time polymerase chain reaction analyses for transglycosylase and superoxide dismutase confirmed that these proteins were up‐regulated under inducing condition, consistent with the proteomic results. Multiple spots for the 60‐kDa chaperonin and glyceraldehyde‐3‐phosphate dehydrogenase were identified, suggesting the presence of post‐translational modifications. We propose that substantial alterations in cellular envelope metabolism occur during the XAC infectious process, which are related to several aspects, from defence against reactive oxygen species to exopolysaccharide synthesis. Our results provide new candidates for virulence‐related proteins, whose abundance correlates with the induction of pathogenicity and virulence genes, such as hrpD6, hrpG, hrpB7, hpa1 and hrpX. The results present new potential targets against XAC to be investigated in further functional studies.     

A novel <Emphasis Type="Italic">PAX5</Emphasis> rearrangement in <Emphasis Type="Italic">TCF3-PBX1</Emphasis> acute lymphoblastic leukemia: a case report

Background

Chromosome translocations are a hallmark of B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Additional genomic aberrations are also crucial in both BCP-ALL leukemogenesis and treatment management. Herein, we report the phenotypic and molecular cytogenetic characterization of an extremely rare case of BCP-ALL harboring two concomitant leukemia-associated chromosome translocations: t(1;19)(q23;q13.3) and t(9;17)(p13;q11.2). Of note, we described a new rearrangement between exon 6 of PAX5 and a 17q11.2 region, where intron 3 of SPECC1 is located. This rearrangement seems to disrupt PAX5 similarly to a PAX5 deletion. Furthermore, a distinct karyotype between diagnosis and relapse samples was observed, disclosing a complex clonal evolution during leukemia progression.

Case presentation

A 16-year-old boy was admitted febrile with abdominal and joint pain. At clinical investigation, he presented with anemia, splenomegaly, low white blood cell count and 92% lymphoblast. He was diagnosed with pre-B ALL and treated according to high risk GBTLI-ALL2009. Twelve months after complete remission, he developed a relapse in consequence of a high central nervous system and bone marrow infiltration, and unfortunately died.

Conclusions

To our knowledge, this is the first report of a rearrangement between PAX5 and SPECC1. The presence of TCF3-PBX1 and PAX5-rearrangement at diagnosis and relapse indicates that both might have participated in the malignant transformation disease maintenance and dismal outcome.

     

ABUNDANCE OF IRRAWADDY DOLPHINS (ORCAELLA BREVIROSTRIS) AND GANGES RIVER DOLPHINS (PLATANISTA GANGETICA GANGETICA) ESTIMATED USING CONCURRENT COUNTS MADE BY INDEPENDENT TEAMS IN WATERWAYS OF THE SUNDARBANS MANGROVE FOREST IN BANGLADESH

Independent observer teams made concurrent counts of Irrawaddy dolphins Orcaella brevirostris and Ganges River dolphins Platanista gangetica gangetica in mangrove channels of the Sundarbans Delta in Bangladesh. These counts were corrected for missed groups using mark-recapture models. For Irrawaddy dolphins, a stratified Lincoln-Petersen model, which incorporated group size and sighting conditions as covariates, and a Huggins conditional likelihood model, which averaged models that individually incorporated group size, sighting conditions, and channel width as covariates, generated abundance estimates of 397 individuals (CV = 10.2%) and 451 individuals (CV = 9.6%), respectively. For Ganges River dolphins, a stratified Lincoln-Petersen model, which incorporated group size as a covariate, and a Huggins conditional likelihood model, which averaged the same models described above, generated abundance estimates of 196 individuals (CV = 12.7%) and 225 individuals (CV = 12.6%), respectively. Although the estimates for both models were relatively close, the analytical advantages of the Huggins models probably outweigh those of the Lincoln-Petersen models. However, the latter should be considered appropriate when simplicity is a priority. This study found that waterways of the Sundarbans support significant numbers of Irrawaddy and Ganges River dolphins, especially compared to other areas where the species have been surveyed.     

Plant regeneration in <Emphasis Type="Italic">Arachis stenosperma</Emphasis> Krapov. and W. C. Gregory from roots and calluses derived from leaflets of <Emphasis Type="Italic">in vitro</Emphasis> plants

Seed explants of A. stenosperma were cultured on MS medium supplemented with 6-benzylaminopurine with the aim of rescuing nonviable accessions stored in seed bank conditions. The regeneration potential of leaf explants from in vitro plants derived from embryonic axes was studied by using whole leaflets and leaflet segments. Explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzylaminopurine and naphthalene acetic acid. Indirect organogenesis was observed in response to 6-benzylaminopurine, either alone or in association with naphthalene acetic acid, in both explant types. Media supplemented with naphthalene acetic acid as the sole growth regulator induced rhizogenesis in whole leaflets and leaflet segments, with subsequent shoot production directly from the roots.     

Phototherapy increases DNA damage in lymphocytes of hyperbilirubinemic neonates

Phototherapy is commonly used in the treatment of hyperbilirubinemia in newborns. No serious side effects related to phototherapy have been observed, but concerns regarding its potential to damage DNA have been expressed, based on animal or cell-culture studies. The aim of this study was to investigate, in neonates with hyperbilirubinemia, the possible relation between phototherapy and DNA damage. The study included 33 full-term newborns with non-physiological jaundice and 14 healthy newborns with physiological jaundice as controls. Phototherapy was performed with an array of six fluorescent lamps producing radiation with wavelengths of 480–520 nm at 12 μW/cm²/nm. DNA damage in lymphocytes was determined by use of the alkaline comet assay. The DNA damage increased significantly with the duration of phototherapy, as shown by measurements at 24, 48, and 72 h (P < 0.001). These findings indicate that phototherapy, widely used in neonatology units, increases DNA damage in newborns. It remains to be seen whether the genotoxic effect observed in the present study can cause any long-term health effect in phototherapy-treated infants in later life.     

SNUFER: A software for localization and presentation of single nucleotide polymorphisms using a Clustal multiple sequence alignment output file

SNUFER is a software for the automatic localization and generation of tables used for the presentation of single nucleotide polymorphisms (SNPs). After input of a fasta file containing the sequences to be analyzed, a multiple sequence alignment is generated using ClustalW ran inside SNUFER. The ClustalW output file is then used to generate a table which displays the SNPs detected in the aligned sequences and their degree of similarity. This table can be exported to Microsoft Word, Microsoft Excel or as a single text file, permitting further editing for publication. The software was written using Delphi 7 for programming and FireBird 2.0 for sequence database management. It is freely available for noncommercial use and can be downloaded from http://www.heranza.com.br/bioinformatica2.htm.     

Thrombin-Induced Regulation of CD95(Fas) Expression in the N9 Microglial Cell Line: Evidence for Involvement of Proteinase-Activated Receptor<Subscript>1</Subscript> and Extracellular Signal-Regulated Kinase 1/2

Microglia are the immune cells of the CNS. Brain injury triggers phenotypic changes in microglia including regulation of surface antigens. The serine proteinase α-thrombin can induce profound changes in neural cell physiology via cleavage of proteinase-activated receptors (PARs). We recently demonstrated that pharmaceutical-grade recombinant human α-thrombin (rh-thr) induces a restricted set of proteolysis-dependent changes in microglia. CD95(Fas) is a cell-death receptor that is up-regulated in microglia by inflammatory stimuli. Here we characterized the effect of rh-thr on CD95(Fas) expression in the N9 microglial cell line. Dose–response and time course studies demonstrated maximal effects at 100 U/ml and 24 h, respectively. Regulation of expression was seen at both the surface protein and steady-state mRNA levels. The rh-thr-induced effects were mimicked by PAR₁ agonist peptides and blocked by pharmacologic inhibitors selective for extracellular signal-regulated kinase 1/2 (ERK 1/2). Rh-thr also induced a rapid and sustained phosphorylation of ERK 1/2. Thrombin-induced regulation of CD95(Fas) could modulate the neuroinflammatory response in a variety of neurological disorders.