COMPATIBILIDADE DE METARHIZIUM ANISOPLIAE COM CARRAPATICIDAS QUÍMICOS

ABSTRACT The combined use of chemical and biological control agents could prove to be a successful strategy against insects and ticks, which could improve the efficiency attained by conventional strategies and also lessen environmental impact that could be caused by these substances. The present study assessed compatibility of the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin to chemical substances currently used for controlling Riphicephalus (Boophilus. microplus (Canestrini), to verify compatibility of these for possible associated use. The fungus was cultivated in a medium to which was incorporated 40, 55, 70, 85 and 100% of the recommended dosage (RD) of commercial acaricides containing various active ingredients formulated in different compositions. To verify compatibility, fungus vegetative growth, spore production and viability were assessed, and all acaricides were classified regarding toxicity. The chemical acaricides containing chlorfenvinphos + dichlorvos (Amiphós®), cipermetrin + chlorpiriphos + citronelal (Barrage®) and amitraz + chlorpiriphos (Carbeson®) were classified as toxic after causing inhibition of fungal growth and spore production, the last one being the only agent that did not affect fungal viability. The acaricide containing cipermetrin (Colosso®) was classified as compatible and moderately compatible in 40 and 65% of the RD, respectively, and the acaricide containing spinosad (Elector®) was the agent that least affected the factors evaluated, being classified as moderately compatible at 55 to 100% of the RD, and as compatible at 40% of RD. Most of the chemical acaricides evaluated were not compatible with M. anisopliae. The acaricide formulated with cipermetrin and spinosad was the only one that showed some compatibility for use in an integrated pest management.

RESUMO A associação de agentes biológicos e químicos pode ser uma estratégia promissora para o controle de insetos e ácaros, visando a aumentar a eficiência, reduzir custos e impactos ambientais. Este trabalho objetivou avaliar a compatibilidade do fungo Metarhizium anisopliae (Metsch.) Sorokin com carrapaticidas químicos atualmente usados no controle de Riphicephalus (Boophilus. microplus (Canestrini). O fungo foi cultivado em meio contendo 40, 55, 70, 85 e 100% da dose recomendada (DR) de carrapaticidas comerciais contendo vários princípios ativos, formulados em diferentes composições. Avaliou-se o crescimento vegetativo, a esporulação e a viabilidade do fungo e fez-se a classificação toxicológica dos acaricidas. Os carrapaticidas contendo clorfenvinfós + diclorvós (Amiphós®), cipermetrina + clorpirifós + citronelal (Barrage®) e amitraz + clorpirifós (Carbeson®) foram classificados como tóxicos, pois inibiram o crescimento e a esporulação, e apenas o último não afetou a viabilidade do fungo. O carrapaticida contendo cipermetrina (Colosso®) foi classificado como compatível e moderadamente compatível nas doses de 40 e 65% da DR, respectivamente, e aquele contendo spinosad (Elector®) foi o que menos afetou os parâmetros avaliados, sendo moderadamente compatível nas doses de 55 a 100% da DR e compatível na dose de 40% da DR. A maioria dos carrapaticidas químicos avaliados não foi compatível com o fungo M. anisopliae; apenas aqueles formulados com cipermetrina e com spinosad mostraram alguma compatibilidade para serem empregados em uma possível estratégia de uso associado.

Compatibilidade de Metarhizium anisopliae com carrapaticidas químicos / Compatibility of Metarhizium anisopliae with chemical acaricides

A associação de agentes biológicos e químicos pode ser uma estratégia promissora para o controle de insetos e ácaros, visando a aumentar a eficiência, reduzir custos e impactos ambientais. Este trabalho objetivou avaliar a compatibilidade do fungo Metarhizium anisopliae (Metsch.) Sorokin com carrapaticidas químicos atualmente usados no controle de Riphicephalus (Boophilus. microplus (Canestrini). O fungo foi cultivado em meio contendo 40, 55, 70, 85 e 100% da dose recomendada (DR) de carrapaticidas comerciais contendo vários princípios ativos, formulados em diferentes composições. Avaliou-se o crescimento vegetativo, a esporulação e a viabilidade do fungo e fez-se a classificação toxicológica dos acaricidas. Os carrapaticidas contendo clorfenvinfós + diclorvós (Amiphós®), cipermetrina + clorpirifós + citronelal (Barrage®) e amitraz + clorpirifós (Carbeson®) foram classificados como tóxicos, pois inibiram o crescimento e a esporulação, e apenas o último não afetou a viabilidade do fungo. O carrapaticida contendo cipermetrina (Colosso®) foi classificado como compatível e moderadamente compatível nas doses de 40 e 65% da DR, respectivamente, e aquele contendo spinosad (Elector®) foi o que menos afetou os parâmetros avaliados, sendo moderadamente compatível nas doses de 55 a 100% da DR e compatível na dose de 40% da DR. A maioria dos carrapaticidas químicos avaliados não foi compatível com o fungo M. anisopliae; apenas aqueles formulados com cipermetrina e com spinosad mostraram alguma compatibilidade para serem empregados em uma possível estratégia de uso associado.

The combined use of chemical and biological control agents could prove to be a successful strategy against insects and ticks, which could improve the efficiency attained by conventional strategies and also lessen environmental impact that could be caused by these substances. The present study assessed compatibility of the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin to chemical substances currently used for controlling Riphicephalus (Boophilus. microplus (Canestrini), to verify compatibility of these for possible associated use. The fungus was cultivated in a medium to which was incorporated 40, 55, 70, 85 and 100% of the recommended dosage (RD) of commercial acaricides containing various active ingredients formulated in different compositions. To verify compatibility, fungus vegetative growth, spore production and viability were assessed, and all acaricides were classified regarding toxicity. The chemical acaricides containing chlorfenvinphos + dichlorvos (Amiphós®), cipermetrin + chlorpiriphos + citronelal (Barrage®) and amitraz + chlorpiriphos (Carbeson®) were classified as toxic after causing inhibition of fungal growth and spore production, the last one being the only agent that did not affect fungal viability. The acaricide containing cipermetrin (Colosso®) was classified as compatible and moderately compatible in 40 and 65% of the RD, respectively, and the acaricide containing spinosad (Elector®) was the agent that least affected the factors evaluated, being classified as moderately compatible at 55 to 100% of the RD, and as compatible at 40% of RD. Most of the chemical acaricides evaluated were not compatible with M. anisopliae. The acaricide formulated with cipermetrin and spinosad was the only one that showed some compatibility for use in an integrated pest management.

Eventos externos e internos da infecção de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae / External and internal events of Rhipicephalus sanguineus larvae and nymphs infection by Metarhizium anisopliae

Examinaram-se a adesão, a germinação, a penetração e a colonização de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae, assim como as lesões infringidas pelo fungo nas respectivas fases do ciclo de vida do ácaro. Realizaram-se infecções experimentais em 11 grupos contendo 250 larvas e 11 grupos contendo 75 ninfas de R. sanguineus, por meio de banho, durante três minutos sob agitação manual, em suspensão contendo 10(8) conídios/ml do fungo. Nos grupos-controles, o banho foi realizado usando o veículo da suspensão. Larvas e ninfas foram processadas para um estudo histopatológico e de microscopia eletrônica de varredura nos seguintes tempos após a infecção: uma e 18 horas, e um, dois, três, quatro, cinco, seis, sete, nove e 11 dias. A germinação dos conídios ocorreu em até 18 horas pós-inoculação, e o fungo penetrou nas larvas e ninfas através do tegumento, dois e três dias após a infecção, respectivamente. Após penetração, o fungo invadiu o corpo das larvas e ninfas, promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Lesões significativas não foram observadas. A morte das larvas e ninfas ocorreu no terceiro e quarto dias pós-infecção, e a esporulação do patógeno sobre o cadáver foi iniciada no sexto dia pós-infecção.

The adhesion, germination and colonization of Rhipicephalus sanguineus larvae and nymphs by Metarhizium anisopliae as well as the lesions caused by the fungus were studied. For this purpose, 11 groups of 250 larvae each and 11 groups of 75 nymphs each were bathed during 3 minutes under manual shaking in a 10(8) conidia/ml suspension. Corresponding control groups were bathed only in the suspension vehicle. Ticks were also submitted to both conventional microscopy and scanning eletronmicrocopy analyses at several post-infection periods (1 and 18 hours and 1, 2, 3, 4, 5, 6, 7, 9, and 11 days). Conidial germination occurred in less than 18 hours post-inoculation and the fungus penetration through the tegument into the larvae and nymphs in, respectively, two and three days post-infection. Following penetration, the fungus invaded the body of the ticks and colonized it diffusely without a noticeable predilection for tissue, but no apparent lesions were observed. Death of larvae and nymphs occurred on the 3rd and 4th post-infection days and pathogen sporulation over the dead tick began on the 6th post-infection day.

Eventos externos e internos da infecção de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae / External and internal events of Rhipicephalus sanguineus larvae and nymphs infection by Metarhizium anisopliae

Examinaram-se a adesão, a germinação, a penetração e a colonização de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae, assim como as lesões infringidas pelo fungo nas respectivas fases do ciclo de vida do ácaro. Realizaram-se infecções experimentais em 11 grupos contendo 250 larvas e 11 grupos contendo 75 ninfas de R. sanguineus, por meio de banho, durante três minutos sob agitação manual, em suspensão contendo 10(8) conídios/ml do fungo. Nos grupos-controles, o banho foi realizado usando o veículo da suspensão. Larvas e ninfas foram processadas para um estudo histopatológico e de microscopia eletrônica de varredura nos seguintes tempos após a infecção: uma e 18 horas, e um, dois, três, quatro, cinco, seis, sete, nove e 11 dias. A germinação dos conídios ocorreu em até 18 horas pós-inoculação, e o fungo penetrou nas larvas e ninfas através do tegumento, dois e três dias após a infecção, respectivamente. Após penetração, o fungo invadiu o corpo das larvas e ninfas, promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Lesões significativas não foram observadas. A morte das larvas e ninfas ocorreu no terceiro e quarto dias pós-infecção, e a esporulação do patógeno sobre o cadáver foi iniciada no sexto dia pós-infecção.(AU)

The adhesion, germination and colonization of Rhipicephalus sanguineus larvae and nymphs by Metarhizium anisopliae as well as the lesions caused by the fungus were studied. For this purpose, 11 groups of 250 larvae each and 11 groups of 75 nymphs each were bathed during 3 minutes under manual shaking in a 10(8) conidia/ml suspension. Corresponding control groups were bathed only in the suspension vehicle. Ticks were also submitted to both conventional microscopy and scanning eletronmicrocopy analyses at several post-infection periods (1 and 18 hours and 1, 2, 3, 4, 5, 6, 7, 9, and 11 days). Conidial germination occurred in less than 18 hours post-inoculation and the fungus penetration through the tegument into the larvae and nymphs in, respectively, two and three days post-infection. Following penetration, the fungus invaded the body of the ticks and colonized it diffusely without a noticeable predilection for tissue, but no apparent lesions were observed. Death of larvae and nymphs occurred on the 3rd and 4th post-infection days and pathogen sporulation over the dead tick began on the 6th post-infection day.(AU)

Kininogens coordinate adaptive immunity through the proteolytic release of bradykinin, an endogenous danger signal driving dendritic cell maturation.

Strategically positioned in peripheral tissues, immune sentinel cells sense microbes and/or their shed products through different types of pattern-recognition receptors. Upon secretion, pre-formed pro-inflammatory mediators activate the microvasculature, inducing endothelium/neutrophil adherence and impairing endothelium barrier function. As plasma proteins enter into peripheral tissues, short-lived proinflammatory peptides are rapidly generated by limited proteolysis of complement components and the kininogens (i.e. kinin-precursor proteins). While much emphasis has been placed on the studies of the vascular functions of kinins, their innate effector roles remain virtually unknown. A few years ago, we reported that exogenous bradykinin (BK) potently induces dendritic cell (DC) maturation, driving IL-12-dependent Th1 responses through the activation of G-protein-coupled BK B(2) receptors (B(2)R). The premise that immature DC might sense kinin-releasing pathogens through B(2)R was demonstrated in the subcutaneous mouse model of Trypanosoma cruzi infection. Analysis of the dynamics of parasite-evoked inflammation revealed that activation of TLR2/neutrophils drives the influx of plasma proteins, including kininogens, into peripheral tissues. Once associated to cell surfaces and/or extracellular matrices, the surface-bound kininogens are cleaved by T. cruzi cysteine proteases. Acting as short-lived 'danger' signals, kinins activate DC via B(2)R, converting them into Th1 inducers. Fine tuned control of the extravascular levels of these natural peptide adjuvants is exerted by kinin-degrading metallopeptidases, e.g. Angiotensin converting enzyme (ACE/CD143). In summary, the studies in the subcutaneous model of T. cruzi infection revealed that the peripheral levels of BK, a DC maturation signal, are controlled by TLR2/neutrophils and ACE, respectively characterized as positive and negative modulators of innate/adaptive immunity.

Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes.

Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs.

Pro domain peptide of HGCP-Iv cysteine proteinase inhibits nematode cysteine proteinases

Cysteine proteinases (CPs) are synthesized as zymogens and converted to mature proteinase forms by proteolytic cleavage and release of their pro domain peptides. A cDNA encoding a papain-like CP, called hgcp-Iv, was isolated from a Heterodera glycines J2 cDNA library, expressed and utilized to assess the ability of its propeptide to inhibit proteinase in its active form. The hgcp-Iv cDNA sequence encodes a polypeptide of 374 amino acids with the same domain organization as other cathepsin L-like CPs, including a hydrophobic signal sequence and a pro domain region. HGCP-Iv, produced in Escherichia coli as a fusion protein with thioredoxin, degrades the synthetic peptide benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin and is inhibited by E-64, a substrate and inhibitor commonly used for functional characterization of CPs. Recombinant propeptides of HGCP-Iv, expressed in E. coli, presented high inhibitory activity in vitro towards its cognate enzyme and proteinase activity of Meloidogyne incognita females, suggesting its usefulness in inhibiting nematode CPs in biological systems. Cysteine proteinases from other species produced no noticeable activity.

Characterization of principal nutritional components of Brazilian oil palm (Eliaes guineensis) fruits.

In spite of the fact that most of the members of Palmaceae contain high concentrations of oil, its potential as a source of oil and protein for human consumption has not been exploited. The pulp and kernels of the Eliaes guineensis palm fruits grown in the Northeast region of Brazil were analyzed only for their proximate composition. The lipid content of the dried pulp and kernels was 73.2% and 32.6%, respectively. Hexane extracted oils from the pulp and kernels yielded similar refractive indices, specific gravity but different peroxide, acid, iodine and saponification values. Gas chromatographic analysis revealed the presence of 24 and 18 fatty acids in pulp and kernel oils, respectively. The principal saturated acid of the pulp oil was palmitic acid (36.9% of the total), and lauric acid (53.3%) for kernel oil. Oleic acid was the predominant monounsaturated fatty acid in both the oils though its concentration in the pulp and kernel oils was 45.29% and 5.5%, respectively. In relation to the essential amino acids, pulp proteins presented a better profile than the kernel proteins. In comparison to the FAO reference protein, the pulp proteins were deficient in methionine, lysine and threonine (16.8%, 51.6% and 93.5% of FAO reference protein) but contained leucine, valine, isoleucine and phenylalanine in optimal concentrations. With exception to phenylalanine and valine (102.2% and 111.4% of reference protein, respectively), the kernel proteins were deficient in all other essential amino acids. The oils from this palm can be used as culinary oil and in margarine manufacture, while pulp could be a supplement for essential amino acids leucine, isoleucine, phenylalanine and valine with other protein sources that are deficient in these amino acids.

Environmental and genetic factors affecting mutability to aminoglycoside antibiotics among Escherichia coli K12 strains

Environmental and genetic factors affecting the in vitro spontaneous mutation frequencies to aminoglycoside resistance in Escherichia coli K12 were investigated. Spontaneous mutation frequencies to kanamycin resistance were at least 100 fold higher on modified Luria agar (L2) plates, when compared to results obtained in experiments carried out with Nutrient agar (NA) plates. In contrast to rifampincin, the increased mutability to kanamycin resistance could not be attributed to a mutator phenotype expressed by DNA repair defective strains. Kanamycin mutant selection windows and mutant preventive concentrations on L2 plates were at least fourfold higher than on NA plates, further demonstrating the role of growth medium composition on the mutability to aminoglycosides. Mutability to kanamycin resistance was increased following addition of sorbitol, suggesting that osmolarity is involved on the spontaneous mutability of E. coli K12 strains to aminoglycosides. The spontaneous mutation rates to kanamycin resistance on both L2 and NA plates were strictly associated with the selective antibiotic concentrations. Moreover, mutants selected at different antibiotic concentrations expressed heterogeneous resistance levels to kanamycin and most of them expressing multiple resistance to all tested aminoglycoside antibiotics (gentamicin, neomycin, amykacin and tobramycin). These results will contribute to a better understanding of the complex nature of aminoglycoside resistance and the emergence of spontaneous resistant mutants among E. coli K12 strains

Identification, characterization and localization of chagasin, a tight-binding cysteine protease inhibitor in Trypanosoma cruzi.

Lysosomal cysteine proteases from mammalian cells and plants are regulated by endogenous tight-binding inhibitors from the cystatin superfamily. The presence of cystatin-like inhibitors in lower eukaryotes such as protozoan parasites has not yet been demonstrated, although these cells express large quantities of cysteine proteases and may also count on endogenous inhibitors to regulate cellular proteolysis. Trypanosoma cruzi, the causative agent of Chagas' heart disease, is a relevant model to explore this possibility because these intracellular parasites rely on their major lysosomal cysteine protease (cruzipain) to invade and multiply in mammalian host cells. Here we report the isolation, biochemical characterization, developmental stage distribution and subcellular localization of chagasin, an endogenous cysteine protease inhibitor in T. cruzi. We used high temperature induced denaturation to isolate a heat-stable cruzipain-binding protein (apparent molecular mass, 12 kDa) from epimastigote lysates. This protein was subsequently characterized as a tight-binding and reversible inhibitor of papain-like cysteine proteases. Immunoblotting indicated that the expression of chagasin is developmentally regulated and inversely correlated with that of cruzipain. Gold-labeled antibodies localized chagasin to the flagellar pocket and cytoplasmic vesicles of trypomastigotes and to the cell surface of amastigotes. Binding assays performed by probing living parasites with fluorescein (FITC)-cruzipain or FITC-chagasin revealed the presence of both inhibitor and protease at the cell surface of amastigotes. The intersection of chagasin and cruzipain trafficking pathways may represent a checkpoint for downstream regulation of proteolysis in trypanosomatid protozoa.

On-line mercury and methylmercury pre-concentration by adsorption of their dithiophosphoric acid diacyl ester chelates on a C18 column and cold-vapor atomic-absorption detection.

The pre-concentration of mercury(II) and methylmercury by adsorption of their dithiophosphoric acid diacyl ester (DDTP) chelates on a C18 column, then detection with cold-vapor atomic-absorption spectrometry was investigated. Conditions such as sample pH, reductant and chelating agent flow and concentration, and eluent and carrier gas flow were optimized. Optimization was performed by use of evolutionary operation with a proper factorial design. At a sample flow of 5.3 mL min(-1) and a loading time of 4.5 min, column adsorption efficiency ranged from 88 to 93% for both species. Detection limits down to 10 ng L(-1) were obtained at a sample throughput of 12 h(-1). There was good agreement between found and certified values in the analysis of certified reference materials after their microwave-assisted mineralization with HNO3 and H2O2.

The protease inhibitor chagasin of Trypanosoma cruzi adopts an immunoglobulin-type fold and may have arisen by horizontal gene transfer.

Chagasin, a protein from Trypanosoma cruzi, is the first member of a new family of tight binding cysteine protease inhibitors [Monteiro, A.C.S., Abrahamson, M., Lima, A.P.C., Vannier-Santos, M.A. and Scharfstein, J. (2001) J. Cell Sci., in press] [corrected]. Despite its lack of significant sequence identity with known proteins, convincing structural models, using variable light chain templates, could be constructed on the basis of threading results. Experimental support for the final structure came from inhibition data for overlapping oligopeptides spanning the chagasin sequence. Chagasin therefore exemplifies a new protease inhibitor structural class and a new natural use for an immunoglobulin-like domain. Limited sequence resemblance suggests that chagasin may represent the result of a rare horizontal gene transfer from host to parasite.

Histoplasmosis of the thyroid.

Fungal infection of the thyroid is rare. Most reported cases have involved Aspergillus, Coccidioides, and Candida species in the setting of disseminated disease. Infection of the thyroid with Histoplasma capsulatum is rarely reported as part of disseminated disease, even in geographic areas where histoplasmosis is endemic. We report a 52-year-old woman with a previous Hashimoto's disease and non-Hodgkin's lymphoma in which a diffuse enlarged thyroid gland with a large nodule was the only apparent locus of histoplasmosis. Fine-needle aspiration of the thyroid was an important diagnostic tool in establishing the diagnosis of histoplasmosis of the thyroid. The patient was initially treated with itraconazole (400 mg/day) for the fungal infection and six cycles of chemotherapy for the lymphoma. At a 6-month follow-up examination, the patient was doing well on suppressive therapy of itraconazole (200 mg/day), with no symptoms and with regression of the thyroid nodule and cervical adenopathy.

Validity of behavioral and physiologic parameters for acute pain assessment of term newborn infants.

CONTEXT: The subjectivity of pain causes enormous difficulties in evaluating neonatal pain with a single, practical and easy-to-apply tool. Pain evaluation in the neonatal period should be performed by valid, safe, useful and feasible methods. OBJECTIVE: To evaluate the validity of the Neonatal Facial Coding System (NFCS), Neonatal Infant Pain Scale (NIPS), heart rate (HR) and O2 saturation (O2 sat) for neonatal pain assessment. DESIGN: Prospective, double-blind randomized trial. SETTING: A secondary level maternity hospital. PARTICIPANTS: 70 healthy neonates requiring bilirubin dosage were randomly assigned to receive a venous puncture (P: n = 33, BW 3.2 kg, SD 0.6; GA 39 wk, SD 1; 59 h of life, SD 25) or an alcohol swab friction (F: n = 37; BW 3.1 kg, SD 0.5; GA 39 wk, SD 1; 52 h of life, SD 17). INTERVENTION: All measurements were taken prior to (PRE), during (TO), and 1 (T1), 3(T3), 5(T5) and 10(T10) minutes after the procedure. MEASUREMENTS: A neonatologist evaluated NFCS, NIPS, HR and O2 sat by pulse oxymetry. RESULTS: Median NFCS and NIPS results at T0, T1 and T3 were higher in P group, compared to F. More P neonates presented NFCS > 2 and/or NIPS > 3 at T0, T1 and T3. HR was lower in P group at T1. Average O2 sat was above 90% during the whole study period in both groups. CONCLUSION: NFCS and NIPS are suitable instruments for neonatal pain evaluation. Heart rate and O2 saturation can be used only as auxiliary methods.

Vulvar myiasis.

Myiasis is a parasitic infestation caused by larvae of several fly species. Diagnosis and treatment are simple. This infestation is, however, rarely seen in the vulvar area. We present a short review of the disease and the case of a 19-year-old pregnant girl with vulvar myiasis and concomitant syphilis, vaginal trichomoniasis and genital candidiasis. The patient was also positive for human immunodeficiency virus.

Isolation and partial characterization of a lectin from Artocarpus incisa L. seeds.

A lectin was isolated from the saline extract of Artocarpus incisa seed by affinity chromatography on cross-linked Adenanthera pavonina galactomannan in 0.15 M NaCl. The lectin was also retained in a D-gal-agarose resin and had no requirements for divalent metal cations (Ca2+ and Mn2+) for activity. The lectin contains 2.1% of carbohydrate and is characterized by high contents of acidic and hydroxylated amino acids. The lectin presented two protein bands in SDS-PAGE, with M(r) 15.5 and 12 kDa, respectively, and contains no alpha-helix, 64% antiparallel beta-sheet and 21% parallel beta-sheet/beta-turn. When submitted to gel filtration in Superose 12 R (FPLC) and Superdex 75 HR 5/5 (HPLC) columns, the lectin showed an M(r) of 48-49 kDa, suggesting a tetrameric structure.

Miíase vulvar / Vulvar myiasis

A miíase é uma infecção parasitária causada por larvas de diversos tipos de moscas. Embora o reconhecimento e tratamento sejam fáceis, constituem uma infecção pouco freqúente na região vulvar. Este estudo apresenta uma revisão a respeito da patologia por miíase e a descrição do caso de uma adolescente de 19 anos, gestante, portadora de miíase vulvar associada à tricomoníase, candidíase e sífilis, além se ser soropositiva para HIV.

[Posttraumatic chylothorax. Experience in the postoperative period following cardiothoracic surgery]. / Quilotórax pós-traumático. Experiência no pós-operatório de cirurgia cardiotorácica.

PURPOSE: To report the accumulated experience in the treatment of patients with postoperative chylothorax (CHT) recovery utilizing pleural drainage associated to alipoidic diet and/or intravenous nutrition. METHODS: The aim of this work is to analyse the management of 11 patients (8 males; 11 months to 70 years old) with post-operative CHT. The previous pathologies were: congenital heart disease in 7; coronary insufficiency in 2; pulmonary tumor in 1 and mediastinal tumor in 1. The diagnosis was made up to 2nd postoperative week in 6, up to 4th week in 3 and later in 2 patients. The volume through the drain ranged from 200 to 3200ml/24h (median 636ml/24h). The laboratory diagnosis was made by lipidic presence in pleural effusion. In all patients the clinical management was made by hipo or alipoidic diet. RESULTS: In 7 the response was good with a decrease of drainage progressively. In 4, it was necessary the introduction of intravenous nutrition by the insufficient response and maintenance of drainage. The reoperation was not used and lymph fistula closed in a period until 10 days in 1 patient; until 20 days in 6 and after this in 4. CONCLUSION: In conclusion, the post-operative CHT may be treated by thoracic drainage and alipoidic diet and/or intravenous nutrition with fistula closure in all patients and without need of reoperation.

[Myocardial revascularization after reperfusion in the acute phase of myocardial infarction]. / Revascularização miocárdica após reperfusão na fase aguda do infarto do miocárdio.

PURPOSE: To evaluate the coronary artery bypass grafting (CABG) after successful coronary reperfusion (CR) by streptokinase and/or percutaneous transluminal coronary angioplasty in acute myocardial infarction (AMI). PATIENTS AND METHODS: During 65-months period, 245 patients underwent CR during AMI. In 47 (19.2%) CABG were performed in the acute period due to multi-vessel disease (31%), residual lesion (20%) and post-reperfusion angina (17%). There were two distinct periods: in the first, between Jun/81 and Jun/83, 34 patients underwent CABG, 47 hours average after reperfusion; in the second, between Jul/83 and Nov/86, 13 patients underwent CABG, 7 days average after reperfusion. RESULTS: There were 7 deaths (21%) in the first period and 1 (8%) in the second one. CONCLUSION: After a multifactorial analysis the authors concluded that better results with inferior mortality (p less than 0.05) were obtained when CABG was performed later (mean 7 days) than in the immediate post-reperfusion period (mean 47 hours).