Nota sôbre a dosagem iodométrica da vitamina C nos frutos cítricos

Ascorbic acid was determined in pure aquous solutions and in citrus fruit juices by iodometric, dichlorophenolindophenol and iodate methods. More constant values were obtained with iodate and Tillmans methods. Iodate is preferable owing to the stability of solution and the simplicity of the method. In the analysis of citrus juices the iodate method proposed by Ballentine is very accurate and suitable for routine work (Table I and II). Recovery experiments recorded in Table III show that the results are reproducible. The averages obtained for some fruits are shown in Table IV. Lemon: 45,4 to 67,3; orange: 28,0 to 60,8; lima: 25,2 to 38,2 and mandarine: 32,0 to 59,3. Values expressed in mg per 100 cc. of juice.

Genomic, Epigenomic and Transcriptomic Molecular Characterization of Splenic Marginal Zone Lymphoma Reveals Recurrent Abnormalities in miR-182

Splenic marginal zone lymphoma (SMZL) is a low grade B-cell non-Hodgkin's lymphoma. The molecular pathology of this entity remains poorly understood. To characterise this lymphoma at the molecular level, we performed an integrated analysis of 1) genome wide genetic copy number alterations 2) gene expression profiles and 3) epigenetic DNA methylation profiles.We have previously shown that SMZL is characterised by recurrent alterations of chromosomes 7q, 6q, 3q, 9q and 18; however, gene resolution oligonucleotide array comparative genomic hybridisation did not reveal evidence of cryptic amplification or deletion in these regions. The most frequently lost 7q32 region contains a cluster of miRNAs. qRT-PCR revealed that three of these (miR-182/96/183) show underexpression in SMZL, and miR-182 is somatically mutated in >20% of cases of SMZL, as well as in >20% of cases of follicular lymphoma, and between 5-15% of cases of chronic lymphocytic leukaemia, MALT-lymphoma and hairy cell leukaemia. We conclude that miR-182 is a strong candidate novel tumour suppressor miRNA in lymphoma.The overall gene expression signature of SMZL was found to be strongly distinct fromthose of other lymphomas. Functional analysis of gene expression data revealed SMZL to be characterised by abnormalities in B-cell receptor signalling (especially through the CD19/21-PI3K/AKT pathway) and apoptotic pathways. In addition, genes involved in the response to viral infection appeared upregulated. SMZL shows a unique epigenetic profile, but analysis of differentially methylated genes showed few with methylation related transcriptional deregulation, suggesting that DNA methylation abnormalities are not a critical component of the SMZL malignant phenotype.

On Pomacea sordida (Swainson, 1823) (Prosobranchia, Ampullariidae)

A description of Pomacea sordida (Swainson, 1823) collected in Caxias and Nova Iguaçu, state of Rio de Janeiro, is presented. The shell is globose, heavy, whith greenish or horn-colored periostracum and dark spinal bands; apex subelevated, 4-5 moderately shoudered whorls, increasing rather rapidly and separated by deep suture. Aperture large, moderately round, yellowish or violaceous; lip thick and sometimes dark brown; umbilicus large and deep; operculum corneous and heavy, entirely closing the aperture. Ratios: shell width/shell length = 0.81-0.91 (mean 0.86); aperture length/shell length = 0.66-0.75 (mean 0.70). Testis, spermiduct and penis pouch as in Pomacea lineata (Spix, 1827). Seminal vesicle whitish and bean-shaped. Prostate cylindric and narrow, cream in coloar as the testis. Penis whiplike whith a closed circular spermiduct. Penial sheath elongated and tapered, with its distal tip turned to the right; outer basal gland situated on the left; inner median gland rounded; apical gland elongated and wrinkled. Ovary composed of branched whitish tubules lying superficially on the digestive gland; oviduct and seminal receptacle as in P. lineata; albumen gland yellowish - orange. Vestigial male copulatory apparatus (penis and its sheath) present in all females examined.

New methods for the diagnosis of Babesia bigemina infection

Accurate diagnosis of Babesia bigemina infection, an economically important tick-transmitted protozoan parasite of cattle, is essential in the management of disease control and in epidemiological studies. The currentlyused methods of diagnosis are blood smear examination and serological tests which include agglutination and immunofluorescence tests. These testes have been used the fild but because they lack sensitivity and specificity, never and improved methods of diagnosis are being developed. The quantitative buffy coat (OBC) method, using microhaematocrit tubes and acridine orange staining allows rapid and quicker diagnosis of B. bigemina and other blood parasites compared to light microscopic examination of stained smears. Parasite specific monoclonal antibodies have been used in antigen/antibody capture enzymelinked immunosorbent assays with grater sensitivity and specificity than previously described serological tests. Similary, DNA probes, derived from a repetitive sequence of the B. bigemina genome, offer a method of detecting very small numbers of parasites which are undetectable by conventional microscopy. An extrachromosomal DNA element, present in all the tick-borne protozoan parasites so far tested, provides an accurate means of diferentiating mixed parasite populations in infected animals. These improved methods will greatly facilitate epidemiological studies.

Life cycle of Misumenops pallida (Araneae, Thomisidae)

Postembryonic development of Misumenops pallida (Keyserling) (Araneae, Thomisidae), one of the most abundant predator species in soybean fields in Buenos Aires Province (Argentina) was studied. The life cycle was observed in the laboratory from egg sacs collected in the field, and from egg sacs spined in the laboratory by gravid females collected in the field. Results indicated that instar length and feeding rate increased throughout the life cycle being higher in females than in males. Greater mortality was observed to third and fourth instars decreasing thereafter. These results may contribute to deternmine the efficiency of this species as a natural enemy of insect pests of soybean.

A Redescription of Riggia paranensis Szidat, 1948 (Isopoda, Cymothoidae) Based on Thirty-two Specimens from Curimatid Fish of Rio de Janeiro, Brazil, with an Emendation of the Genus

Riggia paranensis Szidat, 1948 is redescribed on the basis of 30 female and 2 male specimens collected from the pericardial cavities of the curimatid fish Cyphocarax (= Curimata) gilberti (Quoy & Gaimard). The fishes were caught in the Itabapoana River, State of Rio de Janeiro, southeastern Brazil. The presence of" dwarf" males, as reported by Szidat, was verified. The fusion of the pleonites and pleotelson in adult females was also confirmed. The generic diagnosis was emended to include details of the mouthparts and pleopods

Triatoma jurbergi sp. n. do Norte do Estado do Mato Grosso, Brasil (Hemiptera, Reduviidae, Triatominae) com uma Atualização das Sinonímias e outros Táxons

Triatoma jurbergi n. sp. is described based on nine specimens of both sexes deposited in the Rodolfo Carcavallo Collection in the Oswaldo Cruz Institute Entomological Collection. The new species can be separated from the closely related Triatoma guazu Lent & Wygodzinsky, 1979 by several characters. The most important are longer anteocular region; thin and pointed juga; the shape of the eyes without concavity in the posterior edge; much longer second rostral segment, passing the posterior edge of eye; the absence of a ventral longitudinal depression on the abdomen; the general color redish, brown and orange and the male genitalia, mainly in the vesica lightly chitinized and smaller, the phallosome with apical projection and the pointed apex of the endosome process.

Frequency-dependent sexual selection with respect to offspring fitness returns is consistent with predictions from rock-paper-scissors dynamics in the European common lizard

Genetic polymorphism can be maintained over time by negative frequency-dependent (FD) selection induced by Rock-paper-scissors (RPS) social systems. RPS games produce cyclic dynamics, and have been suggested to exist in lizards, insects, isopods, plants, and bacteria. Sexual selection is predicted to accentuate the survival of the future progeny during negative FD survival selection. More specifically, females are predicted to select mates that produce progeny genotypes that exhibit highest survival during survival selection imposed by adult males. However, no empirical evidence demonstrates the existence of FD sexual selection with respect to fitness payoffs of genetic polymorphisms. Here we tested this prediction using the common lizard Zootoca vivipara, a species with three male color morphs (orange, white, yellow) that exhibit morph frequency cycles. In a first step we tested the congruence of the morph frequency change with the predicted change in three independent populations, differing in male color morph frequency and state of the FD morph cycle. Thereafter we ran standardized sexual selection assays in which we excluded alternative mechanisms that potentially induce negative FD selection, and we quantified inter-sexual behavior. The patterns of sexual selection and the observed behavior were in line with context-dependent female mate choice and male behavior played a minor role. Moreover, the strength of the sexual selection was within the magnitude of selection required to produce the observed 3-4-year and 6-8 year morph frequency cycles at low and high altitudes, respectively. In summary, the study provides the first experimental evidence that underpins the crucial assumption of the RPS games suggested to exist in lizards, insects, isopods, and plants; namely, that sexual selection produces negative-FD selection. This indicates that sexual selection, in our study exert by females, might be a crucial driver of the maintenance of genetic polymorphisms.

Description of Pythonella scleruri n. sp. (Apicomplexa: Eimeriidae) from a Brazilian Bird Rufous-Breasted-Leaftosser Sclerurus scansor, 1835 (Passeriformes: Furnariidae)

Coccidian oocysts containing 16 sporocysts with 4 sporozoites in each were observed in a faecal sample from Sclerurus scansor collected in the Itatiaia National Park, southeastern region of Brazil. The oocysts are characterized by ellipsoidal shape measuring 42.5 x 32.8 mm, with smooth, thick double-layered wall of a greenish-orange colour. An oocyst residuum of numerous scattered granules among the sporocysts in sporulated ones; 16 round sporocysts, averaging 10.5 x 10 mm each containing four elongated sporozoites; presence of residuum; absence of Stieda body. The presently described coccidian, recorded for the first time in birds, is a new species named P. scleruri.

The in vitro cytopathology of a porcine and the simian (SA-11) strains of rotavirus

Rotaviruses have been implicated as the major causal agents of acute diarrhoea in mammals and fowls. Experimental rotavirus infection have been associated to a series of sub-cellular pathologic alterations leading to cell lysis which may represent key functions in the pathogenesis of the diarrhoeic disease. The current work describes the cytopathic changes in cultured MA-104 cells infected by a simian (SA-11) and a porcine (1154) rotavirus strains. Trypan blue exclusion staining showed increased cell permeability after infection by both strains, as demonstrated by cell viability. This effect was confirmed by the leakage of infected cells evaluated by chromium release. Nuclear fragmentation was observed by acridine orange and Wright staining but specific DNA cleavage was not detected. Ultrastructural changes, such as chromatin condensation, cytoplasm vacuolisation, and loss of intercellular contact were shown in infected cells for both strains. In situ terminal deoxynucleotidyl transferase (Tunel) assay did not show positive result. In conclusion, we demonstrated that both strains of rotavirus induced necrosis as the major degenerative effect.

Ingestion of saliva during carbohydrate feeding by Lutzomyia longipalpis (Diptera; Psychodidae)

The aim of this study was to obtain experimental evidence that phlebotomine saliva is actually ingested during the carbohydrate ingestion phase (before and after blood digestion). The ingestion of carbohydrate was simulated as it occurs in the field by offering the insects balls of cotton soaked in sucrose, sucrose crystals or orange juice cells. The results obtained here showed that ingestion occurred under each condition investigated, as indicated by the presence of apyrase, an enzyme used as a marker to detect saliva in the insect gut and/or carbohydrate sources. Saliva ingestion by phlebotomine during the carbohydrate ingestion phase is important to explain how it could promote starch digestion and to trigger Leishmania promastigotes to follow a differentiation pathway as proposed previously by some authors.

Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%). When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs); mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

Antimalarial drugs disrupt ion homeostasis in malarial parasites

Plasmodium chabaudi malaria parasite organelles are major elements for ion homeostasis and cellular signaling and also target for antimalarial drugs. By using confocal imaging of intraerythrocytic parasites we demonstrated that the dye acridine orange (AO) is accumulated into P. chabaudi subcellular compartments. The AO could be released from the parasite organelles by collapsing the pH gradient with the K+/H+ ionophore nigericin (20 µM), or by inhibiting the H+-pump with bafilomycin (4 µM). Similarly, in isolated parasites loaded with calcium indicator Fluo 3-AM, bafilomycin caused calcium mobilization of the acidic calcium pool that could also be release with nigericin. Interestingly after complete release of the acidic compartments, addition of thapsigargin at 10 µM was still effective in releasing parasite intracellular calcium stores in parasites at trophozoite stage. The addition of antimalarial drugs chloroquine and artemisinin resulted in AO release from acidic compartments and also affected maintenance of calcium in ER store by using different drug concentrations.

Polymerase chain reaction genotyping of Epstein-Barr virus in scraping samples of the tongue lateral border in HIV-1 seropositive patients

The Epstein-Barr virus (EBV) is the etiological agent of oral hairy leukoplakia (OHL), an oral lesion with important diagnostic and prognostic value in acquired immunodeficiency disease syndrome. The two EBV genotypes, EBV-1 and EBV-2, can be distinguished by divergent gene sequences encoding the EBNA-2, 3A, 3B, and 3C proteins. The purpose of this study was to identify the EBV genotype prevalent in 53 samples of scrapings from the lateral border of the tongue of HIV-1 seropositive patients, with and without OHL, and to correlate the genotypes with presence of clinical or subclinical OHL with the clinic data collected. EBV-1 and EBV-2 were identified through PCR and Nested-PCR based on sequence differences of the EBNA-2 gene. EBV-1 was identified in the 31 samples (15 without OHL, 7 with clinical OHL and 9 with subclinical OHL), EBV-2 in 12 samples (10 without OHL, 1 with clinical and 1 subclinical OHL), and a mixed infection in 10 samples (2 without OHL, 3 with clinical and 5 with subclinical OHL). The presence of EBV-1 was higher in women, but a significant statistical result relating one the EBV genotypes to the development of OHL was not found. We conclude that the oral epithelium in HIV-1 seropositive patients can be infected by EBV-1, EBV-2 or by a mixed viral population.