Native Desmodesmus sp. and Chlorella sp. isolated from the Reconquista River display a different binding preference for Cu(II) and Zn(II).

The present work was developed to study the metal removal performance of unicellular algae isolated from the Reconquista River and to evaluate the effect of the presence of more than one metal in the removal process. Thus, native species of unicellular algae were isolated from the highly contaminated Reconquista River. All of the isolates were classified, at genus level, based on their morphological appearance. Nine isolates were screened for their Zn(II) removal capacities. Chlorella sp. RR5 and Desmodesmus sp. RR7 were selected based on their removal performance, and their potential in the remediation of multiple metals was analyzed. Therefore, zinc (Zn(II)), copper (Cu(II)), and chromium (Cr(VI)) removal was evaluated in mono- and multi-metallic solutions. Biosorption capacities were high (0.8-1.8 mmol g-1) for Zn(II) and Cu(II) in mono-metallic solutions. Removal capacities decreased up to 48% in multi-metallic solutions. Interestingly, when multi-metallic systems were considered, each strain showed a metal preference. Chlorella sp. removed better Cu(II) meanwhile Desmodesmus sp. showed a preference for Zn(II). Thus, a metal-binding selectivity in each strain was determined. Chromium (VI) remediation was almost null in the conditions analyzed in this work. Fourier transformation infrared spectroscopy (FT-IR) analysis showed that polysaccharides were the main functional group involved in metal adsorption and, in some cases, also the carboxylates played an important role. Overall, we were able to analyze a new source of algal diversity and perform a metal removal characterization of them, leading to the identification of a metal selectivity based on the characteristics of the tested algal strains.

Reduced levels of NADH-dependent glutamate dehydrogenase decrease the glutamate content of ripe tomato fruit but have no effect on green fruit or leaves.

Glutamate (Glu) is a taste enhancer that contributes to the characteristic flavour of foods. In fruit of tomato (Solanum lycopersicum L.), the Glu content increases dramatically during the ripening process, becoming the most abundant free amino acid when the fruit become red. There is also a concomitant increase in NADH-dependent glutamate dehydrogenase (GDH) activity during the ripening transition. This enzyme is located in the mitochondria and catalyses the reversible amination of 2-oxoglutarate to Glu. To investigate the potential effect of GDH on Glu metabolism, the abundance of GDH was altered by artificial microRNA technology. Efficient silencing of all the endogenous SlGDH genes was achieved, leading to a dramatic decrease in total GDH activity. This decrease in GDH activity did not lead to any clear morphological or metabolic phenotype in leaves or green fruit. However, red fruit on the transgenic plants showed markedly reduced levels of Glu and a large increase in aspartate, glucose and fructose content in comparison to wild-type fruit. These results suggest that GDH is involved in the synthesis of Glu in tomato fruit during the ripening processes. This contrasts with the biological role ascribed to GDH in many other tissues and species. Overall, these findings suggest that GDH has a major effect on the control of metabolic composition during tomato fruit ripening, but not at other stages of development.

Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil / Espécies de Eimeria em ovinos jovens e adultos criados em sistema intensivo e / ou semi - intensivo de um rebanho da cidade de Umuarama, Estado do Paraná, Brasil

The present study aimed to identify Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil using the traditional diagnostic methods and to correlate the infection level/types of infection in the different age/system in this herd. Fecal samples were collected from the rectum of 210 sheep and were subjected to laboratory analysis to differentiate the species. Furthermore, animals were observed to determine the occurrences of the clinical or subclinical forms of eimeriosis. Out of the 210 collected fecal samples, 147 (70%) were positive for Eimeria oocysts, and 101 (47.86%) belonged to young animals that were raised under intensive and / or semi-intensive farming systems. Oocysts from 9 species of Eimeria parasites were identified in the sheep at the following prevalence rates: E. crandallis, 50.0%; E. parva, 21.6%; E. faurei, 8.1%; E. ahsata, 8.1%; E. intricata, 5.4%; E. granulosa , 2.7%; E. ovinoidalis , 2.0%; E. ovina , 1.3%; and E. bakuensis , 0.6%. There were no differences regarding the more frequent Eimeria species among the different ages of animals or between the different farming management systems. Based on these data, E. crandallis was the most prevalent, followed by E. parva and E. faurei species, regardless of the age. Higher parasitism was diagnosed in the young animals that were raised in a confinement regime, and the disease found in the herd was classified as subclinical. Further studies should be conducted in this herd, to verify if the eimeriosis subclinical can cause damage especially in young animals with a high level of infection.

O presente trabalho teve como objetivo realizar a identificação das espécies de Eimeria em ovinos jovens e adultos, criados em sistema intensivo e/ou semi-intensivo, de um rebanho da cidade de Umuarama, estado do Paraná, Brasil, utilizando-se métodos tradicionais de diagnóstico, bem como correlacionando o grau de infecção/tipo de infecção nas diferentes categorias animal presentes no rebanho. Amostras de fezes de 210 ovinos foram colhidas diretamente da ampola retal desses animais. Em seguida, procedeu-se à diferenciação das espécies em laboratório. Além disso, os animais foram observados, diariamente, durante o período de 20 dias, após a colheita de fezes, a fim de se evidenciar se a eimeriose ocorreu de forma clínica ou subclínica nos rebanhos. Das 210 amostras de fezes colhidas, 147 (70%) foram positivas para oocistos de Eimeria. Dentre as 210 amostras, 101 (47,8%) pertenciam aos animais jovens criados no sistema intensivo e/ou semi-intensivo. Foram identificados oocistos de nove espécies de Eimeria parasitas de ovinos, com a seguinte prevalência: E. crandallis (50,0%), E. parva (21,6%), E. faurei (8,1%), E. ahsata (8,1%), E. intrincata (5,4%), E. granulosa (2,7%), E. ovinoidalis (2,0%), E. ovina (1,3%) e E. bakuensis (0,7%). Não houve diferença quanto às espécies de Eimeria mais frequentes, entre as diferentes idades dos animais e também entre os diferentes regimes de criação (manejo). Diante do exposto, E. crandallis foi a espécies mais prevalente, seguida da E. parva e E. faurei, independente da idade dos animais. Maior nível de parasitismo foi encontrado nos animais jovens criados em regime de confinamento, e a enfermidade foi classificada como eimeriose subclínica, independente da idade dos animais. Futuros estudos deverão ser conduzidos neste rebanho, a fim de evidenciar se a eimeriose subclínica desencadeia danos aos animais, especialmente nos jovens criados em confinamento.

Novel glutamate dehydrogenase genes show increased transcript and protein abundances in mature tomato fruits.

NAD(P)H-glutamate dehydrogenase (GDH, EC 1.4.1.3) contributes to the control of glutamate homeostasis in all living organisms. In bacteria and animals, GDH is a homohexamer allosterically regulated, whereas in plants NADH-GDH (EC 1.4.1.2) is also found as heterohexamer of α- and ß-subunits, but its regulation remains undefined. In tomato (Solanum lycopersicum), GDH activity increases during the fruit ripening along with the content of free glutamate, the most abundant amino acid of ripe fruit involved in conferring the genuine tomato flavour. In this work, novel Slgdh-NAD genes were identified in the recently deciphered tomato genome: three encoding the α-subunit (Slgdh-NAD;A1-3) and one additional gene encoding the ß-subunit of GDH (Slgdh-NAD;B1) isolated from a genomic library. These genes are located in different chromosomes. Slgdh-NAD;A1-3 show conserved structures, whereas Slgdh-NAD;B1 includes a novel 5'-untranslated exon. Slgdh-NAD;A1-3 transcripts were detected in all tomato tissues examined, showing the highest levels in mature green fruits, contrasting with Slgdh-NAD;B1 transcripts which were detected mainly in roots or in mature fruits when treated with glutamate, NaCl or salicylic acid. Analyses of GDH activity and protein distribution in different tissues of the Micro-Tom cultivar showed that only the active homohexamer of GDH ß-subunits was detected in roots while heterohexamers of GDH α- and ß-subunits were found in fruits. These results indicate that GDH ß-subunit could modulate the heteromeric isoforms of GDH in response to the environment and physiology of the tomato fruit. This information is relevant to manipulate glutamate contents in tomato fruits genetically.

Alterações eletromiográficas e histopatológicas da musculatura de cães naturalmente infectados por Leishmania infantum / Muscular electromyographic and histopathologic changes in dogs naturally infected by Leishmania infantum

O objetivo do presente estudo foi avaliar as alterações eletromiográficas e histopatológicas de músculos estriados esqueléticos de cães naturalmente infectados por Leishmania infantum. Foram selecionados 25 cães adultos, sem raça definida, com diagnósticos parasitológico, molecular e sorológico estabelecidos para a infecção. Os músculos avaliados foram: tríceps braquial, extensor carpo radial, bíceps femoral e gastrocnêmio. Um cão possuía problemas locomotores, com paresia de membros posteriores associada à intensa atrofia muscular. Vinte e três (92%) apresentavam algum tipo de alteração muscular, sendo que em 22 (88%) tais alterações foram identificadas diretamente pela eletromiografia. Mesmo sem sinais clínicos, em dez cães (40%) foram evidenciadas alterações eletromiográficas e histopatológicas. Antígenos de Leishmania foram identificados na musculatura de quatro (16%) cães. Os resultados eletromiográficos indicaram a ocorrência de polimiosite crônica em 13 (52%) cães, presença de músculos com inflamação tanto aguda quanto crônica em quatro (16%), miopatia aguda em dois (8%), e ausência de alterações eletromiográficas em três (12%). As alterações histopatológicas mais frequentemente observadas foram degeneração e necrose de miofibras e presença de infiltrado inflamatório verificadas em 12 (48%) cães. Outras alterações, quando comparado com as amostras de cães normais, foram do tamanho de grupos de fibras musculares em 15 (60%) e fibrose peri ou endomisial em 14 (56%) animais. As alterações observadas no presente estudo permitiram concluir que mesmo na ausência de sinais clínicos de comprometimento muscular, a maior parte dos cães infectados por L. infantum apresenta polimiosite crônica.

The aim of this study was to evaluate the electromyographic and histopathological changes in skeletal muscles of dogs naturally infected by L. infantum. Twenty five mixed breed adult dogs with parasitological, molecular and serological diagnosis were selected. The evaluated muscles were: triceps brachial, extensor carpi radialis, biceps femoris and gastrocnemius. One dog had locomotor clinical signs with hind limbs paresis associated with severe muscle atrophy. Twenty-three (92%) had some type of muscular change, and in 22 (88%) such changes were directly identified by electromyography. Even without any clinical signs of the disease, 10 (40%) dogs had electromyographic and histopathological changes. Leishmania antigens were detected in muscles of four (16%) dogs. The electromyographic evaluation indicated the occurrence of chronic polymyositis in 13 (52%) dogs, the presence of both acute and chronic muscle inflammation four (16%), acute myopathy in two (8%) and absence of electromyographic abnormalities in three (12%) dogs. The most frequently observed histopathological changes were degeneration and necrosis of myofibers and inflammatory infiltration observed in 12 (48%) dogs. Other changes were decreased diameter of muscle fibers in 15 (60%) and peri or endomysial fibrosis in 14 (56%) animals. The changes observed in the present study showed that even in the absence of clinical signs, most dogs infected by Leishmania infantum have chronic polymyositis.

Free amino acid production during tomato fruit ripening: a focus on L-glutamate.

In tomato, free amino acids increase dramatically during fruit ripening and their abundance changed differentially. More evident is L-glutamate which gives the characteristic "umami" flavor. Glutamate is the principal free amino acid of ripe fruits of cultivated varieties. In this paper, we examined the capacity of tomato fruits to process endogenous as well as exogenous polypeptides during the ripening transition, in order to analyze their contribution to the free amino acid pool. In addition, the activity of some enzymes involved in glutamate metabolism such as gamma-glutamyl transpeptidase (gamma-GTase), glutamate dehydrogenase (GDH), alpha-ketoglutarate-dependent gamma-aminobutyrate transaminase (GABA-T), alanine and aspartate aminotransferases was evaluated. Results showed that peptidases were very active in ripening fruits, and they were able to release free amino acids from endogenous proteins and glutamate from exogenously added glutamate-containing peptides. In addition, red fruit contained enough gamma-GTase activity to sustain glutamate liberation from endogenous substrates such as glutathione. From all the glutamate metabolizing enzymes, GDH and GABA-T showed the higher increase in activities when the ripening process starts. In summary, tomato fruits increase free amino acid content during ripening, most probably due to the raise of different peptidase activities. However, glutamate level of ripe fruit seems to be mostly related to GDH and GABA-T activities that could contribute to increase L-glutamate level during the ripening transition.

Efficient expression of a Toxoplasma gondii dense granule Gra4 antigen in tobacco leaves.

A His-tagged truncated version of Toxoplasma gondii dense granule 4 protein (Gra4(163-345)) was transiently expressed in tobacco leaves. Two genetic constructions were used to accomplish this goal. In one of them, based in a Potato virus X (PVX) amplicon, the sequence encoding His-Gra4(163-345) was placed under control of an additional PVX coat protein subgenomic promoter. In the other, the same sequence was fused to an apoplastic transport signal and placed under the direction of the cauliflower mosaic virus 35S promoter. His-Gra4(163-345) accumulation in agroinfiltrated tobacco leaves was estimated by Western blot analysis using mouse anti-Gra4 antibody and a seropositive human serum. Here, we demonstrated the feasibility of producing a Gra4 antigen using transient expression methods in plants.

Propriedades mecânicas de três fios de sutura no reparo do tendão do músculo flexor profundo do dedo de cães / Mechanical properties of three threads of suture in the repair of the tendon of the muscle deep digital flexor of dogs

Considerando a complexidade da cicatrização tendínea e os fatores que interferem no fenômeno, o escopo desse trabalho foi o de avaliar comparativamente as diferenças nas propriedades biomecânicas dos materiais usualmente utilizados em tenorrafia. Para tanto tendões dos músculos flexores profundo dos dedos de cães (TMFPD) foram colhidos de nove cães mestiços adultos que vieram à óbito sem diagnóstico de lesões no aparelho locomotor. Os 18 TMFPD dos membros torácicos foram aleatoriamente selecionados em três grupos e submetidos à tenotomia e tenorrafia utilizando fios Prolene®, Nylon® e Vicryl®. Para a sutura do epitendão utilizou-se fios de diâmetro 6.0 e para o endotendão 4.0. Os tendões foram suturados com um desses materiais de sutura utilizando a técnica de Kessler modificada. O estudo foi dividido em dois ensaios: Ensaio1: Os tendões ensaiados foram testados na máquina universal de ensaios biomecânicos e as propriedades avaliadas foram: Limite Máximo (LM), Limite de Elasticidade (LE), Rigidez (RG) e Resiliência (RS). Ensaio2: os fios de mononailon, ácido poliglicólico e polipropileno nos diâmetros 6.0 e 4.0 foram testados isoladamente quanto às propriedades Limite Máximo (LM), Rigidez (RG) e Resiliência (RS). Nas condições em que foi realizado o presente estudo os resultados permitem inferir que embora o Vicryl® tenha apresentado a maior rigidez, mostrou-se o mais apropriado para tenorrafias experimentais em cães, uma vez que em todas as outras propriedades biomecânicas apontou o melhor desempenho nos testes empregados.

Estudo morfológico de tendões flexores de eqüinos / Morphological study of equine flexor tendons

A proposta deste estudo é a de descrever as características morfológicas dos tendões dos músculos flexores superficial (TMFSD) e profundo (TMFPD) dos dedos de eqüinos e focalizar suas diferenças. O material utilizado foi obtido de quatro segmentos tendíneos colhidos dos membros torácicos e pélvicos, de sete cadáveres de eqüinos adultos sem lesões macroscópicas diagnosticadas. Os segmentos examinados do TMFSD foram: junção músculo-tendínea, terço médio do terceiro metacarpo ou metatarso, terço distal do terceiro metacarpo ou metatarso e extremidade distal da falange proximal. Por sua vez, para o TMFPD: junção músculo-tendínea, região do terço médio do terceiro metacarpo ou metatarso, região do ligamento frenador inferior e região da falange média. Os resultados evidenciaram algumas diferenças histológicas na distribuição e localização dos componentes entre segmentos do mesmo tendão e entre tendões torácicos e pélvicos, especialmente para a quantidade de condrócitos, que mostrou ser maior na região do terço médio do terceiro metacarpo ou metatarso, tanto para os membros torácicos como para os pélvicos do TMFSD. No caso do TMFPD, o maior número de tais células foi observado na região do terço médio do terceiro metacarpo dos membros torácicos e na região da falange média dos membros pélvicos

Propriedades mecânicas de três padröes de sutura no reparo de tendäo do músculo flexor profundo do dedo em eqüinos / Mechanical properties of three patterns of suture in the repair of tendon of the muscle deep digital flexor equine

Quatro propriedades mecânicas: Limite Máximo (LM), Limite de Elasticidade (LE), Rigidez (RG) e Resiliência (RS) de três padröes de sutura foram avaliadas por ensaios mecânicos de traçäo. Trinta tendöes dos músculos flexores profundos dos dedos de cadáveres de eqüinos foram separados em três grupos de igual número, aproximados com as suturas de Kessler modificada com sutura no epitendäo (KM), Becker modificada (BM) e dupla laçada (DL). A resistência à formaçäo de espaço e a perda de tensäo também foram avaliadas. Para a carga no LM, somente o padräo de sutura KM mostrou-se significativamente diferente (P<0,05 - KM>DL>BM), enquanto no LE, os três padröes de sutura comportaram-se igualmente (P>0,05 - KM>DL>BM). Quanto à deformaçäo, tanto no LM quanto no LE, os três padröes de sutura apresentaram diferenças significativas entre si (P<0,05 - DL>BM>KM). Na RG somente KM comportou-se diferente (P<0,05 - KM>BM>DL). Na RS, os três padröes de sutura apresentaram diferenças significativas entre si (P<0,05 - DL>BM>KM). Quanto à perda de resistência, o padräo KM falhou por distensäo e perda da funçäo da sutura, enquanto os padröes de sutura BM e DL falharam por ruptura do fio. O padräo de sutura KM mostrou-se superior aos outros dois, cujas propriedades mecânicas foram equivalentes