Tolerance of dark septate endophytic fungi (DSE) to 1 agrochemicals in vitro / Tolerancia de hongos endofíticos septados oscuros a agroquímicos in vitro

Abstract Dark septate endophytes (DSE) are a heterogeneous group of fungi, mostly belonging to the Phylum Ascomycota, that are involved in a mutualistic symbiosis with plant roots. The aim of this study is to evaluate the behavior of two strains of DSE isolated from wheat roots of two cropping areas in the province of Buenos Aires, Argentina, against some agrochemicals. Of all the isolates obtained, two strains were identified as Alternaria alternata and Cochliobolus sp. These DSE were found to be tolerant to glyphosate, carbendazim and cypermethrin when evaluated at the recommended agronomic dose (AD), 2 AD and, in some cases, 10 AD. This work contributes to the study of the biology of this group of fungi and their tolerance in the presence of xenobiotics widely used in agriculture.© 2019 Asociaci´on Argentina de Microbiolog´ıa. Published by Elsevier Espa˜na, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/bync-nd/4.0/).

Resumen Los endófitos septados oscuros (DSE) son un grupo heterogéneo de hongos que participan de una simbiosis mutualista con raíces de plantas, perteneciendo principalmente al Phylum Ascomycota. El objetivo de este estudio fue aislar DSE de raíces de trigo proveniente de dos áreas de cultivo de la provincia de Buenos Aires y evaluar el comportamiento de dos cepas de DSE aisladas de raíces de trigo frente a algunos agroquímicos en dos áreas de cultivo de la provincia de Buenos Aires. De todos los aislamientos obtenidos se seleccionaron dos cepas que se identificaron como Alternaria alternata y Cochliobolus sp. Se encontró que estos DSE son tolerantes al glifosato, el carbendazim y la cipermetrina, evaluados a las dosis agronómicas recomendadas (AD), a 2x AD y, en algunos casos, a 10x AD. Este trabajo contribuye al conocimiento de la biología de este grupo de hongos y su tolerancia a xenobióticos ampliamente utilizados en la agricultura.

Efecto del glifosato sobre la microbiota, calidad del suelo y cultivo de frijol biofortificado en el departamento del Cesar, Colombia / Effect of glyphosate on microbiota, soil quality and biofortified bean crop in Codazzi, department of Cesar, Colombia

Resumen Las estrategias en seguridad alimentaria con cultivos de alto contenido nutricionaldeben enmarcarse en prácticas agrícolas sostenibles, orientadas a la conservación del suelo, elalto rendimiento y la inocuidad. Esta última característica implica la producción de alimentos sintrazas detectables de agroquímicos, los que podrían amenazar la salud del consumidor. Se des-arrolló un estudio con el objetivo de evaluar el efecto del herbicida glifosato sobre la fertilidadquímica y microbiológica del suelo, así como su residualidad en la semilla de frijol biofortificadocultivado en el departamento de Cesar, Colombia. La metodología comprendió un análisis corre-lacional de indicadores de calidad de suelo, rendimiento del cultivo y residualidad en los granos.Los tratamientos evaluados incluyeron la aplicación o no de glifosato, el uso de coberturas sin-téticas (mulch) o naturales, frente al control manual de las malezas. Se hallaron diferenciasde rendimiento y de la respuesta de los indicadores químicos y microbiológicos en función deltratamiento y las condiciones iniciales de la rizósfera de frijol. La aplicación del herbicida glifo-sato en suelo sin mulch generó una disminución del 29% en el rendimiento, asociada a la mayorprevalencia de plagas y enfermedades fúngicas. En ninguno de los tratamientos con aplicaciónde glifosato se observó residualidad de este herbicida en semillas, pero sí de otras moléculasderivadas de insecticidas usados en el sistema local de producción. De este estudio se concluyóque en lotes de frijol biofortificado con alta presión de la maleza Cyperus rotundus (coquito), se recomienda el uso del herbicida glifosato acompañado de mulch como alternativa para mantener la productividad en el tiempo. Esto constituye un sistema de protección frente al efectonegativo del herbicida sobre ambos, el sistema radical de la planta y la microbiota del suelo.© 2019 Asociacion Argentina de Microbiologıa. Publicado por Elsevier España, S.L.U. Este es un artıculo Open Access bajo la licencia CC BY-NC-ND (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Abstract Strategies aimed at achieving food safety in crops of high nutritional value shouldbe carried out through sustainable agricultural practices aimed at soil conservation, high yieldand food safety. This latter characteristic implies food production without detectable tracesof agrochemicals that threaten health. The objective of this study was to assess the effect ofthe herbicide glyphosate on the chemical and biological fertility of the soil and to determineits residual activity on biofortified bean seeds in Codazzi, Department of Cesar, Colombia. Themethod included a correlational analysis of soil quality, crop yield and residuality in bean grains.The treatments included glyphosate application and synthetic and natural mulches, comparedto manual control. The results showed differences in the response of chemical, microbiologicaland yield indicators between treatments and the initial conditions of the bean rhizosphere. Theuse of the herbicide glyphosate in mulch-free soil generates yield losses of 29% associated witha higher incidence of pests and fungal diseases; in all treatments, no glyphosate residualitywas detected in seeds; however, residuality was detected in other molecules derived frominsecticides associated with the local production system. In conclusion, in plots with high weedpressure by Cyperus rotundus, the use of mulch is recommended as an alternative to maintainbean productivity over time; by offering protection against the effect of the herbicide on boththe root system of the plant and the soil microbiota of the biofortified bean crop.© 2019 Asociaci´on Argentina de Microbiolog´ıa. Published by Elsevier España, S.L.U. This is anopen access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Morphological effects on helminth parasites caused by herbicide under experimental conditions / Efeitos morfológicos em helminto parasito causado por herbicida em condições experimentais

Abstract Helminth parasites have been studied as potential accumulators for different pollutants. Echinostoma paraensei is a foodborne trematode whose vertebrate host, the rodent Nectomys squamipes, is naturally exposed to environmental pesticides. However, little information exists regarding the pesticide's effects on helminths. This study investigated the morphological effects on the trematode, E. paraensei, after experimental Roundup® herbicide exposure, in concentrations below those recommended for agricultural use. After two hours of exposure, scanning electron microscopy (SEM) showed changes to the tegument, such as furrowing, shrinkage, peeling, spines loss on the peristomic collar, and histopathological evidence of altered cells in the cecum and acinus vitelline glands with vacuoles and structural changes to the muscular layers. Glycidic content was decreased, primarily in the connective tissue. As E. paraensei is an intestinal parasite of the semi-aquatic wild rodent, N. squamipes, it is predisposed to pesticide exposure resulting from agricultural practices. Therefore, we emphasize the need to evaluate its impact on helminth parasites, due to their pivotal role in regulating host populations.

Resumo Helmintos parasitos tem sido estudados como acumuladores potenciais para diferentes poluentes. O trematódeo E. paraensei tem como hospedeiro vertebrado o roedor Nectomys squamipes naturalmente exposto a pesticidas no meio ambiente. No entanto, pouca informação está disponível sobre os efeitos dos pesticidas em helmintos parasitos. O presente estudo investigou, em condições experimentais, os efeitos morfológicos no trematódeo E. paraensei após a exposição ao herbicida Roundup®, em concentrações abaixo das recomendadas para a utilização agrícola. A microscopia eletrônica de varredura (MEV) mostrou após duas horas de exposição, alterações no tegumento, como enrugamento, contração e descamação com perda de espinhos no colar peristômico e análise histopatológica evidenciou células do ceco alteradas, as glândulas vitelínicas com vacúolos e mudanças estruturais nas camadas musculares. Diminuição do conteúdo glicídico, principalmente no tecido conjuntivo, também foi observado. Considerando a predisposição à exposição a pesticidas agrícolas de N. squamipes infectado por E. paraensei, são necessários estudos para avaliar o impacto de tais resíduos frente aos helmintos e seus hospedeiros.

Thidiazuron (TDZ) increases fruit set and yield of 'Hosui' and 'Packham's Triumph' pear trees

ABSTRACT The low fruit set is one of the main factors leading to poor yield of pear orchards in Brazil. The exogenous application of thidiazuron (TDZ) and aminoethoxyvinilglycine (AVG) has shown promising results in some pear cultivars and other temperate fruit trees. The objective of this study was to evaluate the effect of TDZ and AVG on fruit set, yield, and fruit quality of 'Hosui' and 'Packham's Triumph' pears. The study was performed in a commercial orchard located in São Joaquim, SC. Plant material consisted of 'Hosui' and 'Packham's Triumph' pear trees grafted on Pyrus calleryana. Treatments consisted on different rates of TDZ (0 mg L-1, 20 mg L-1, 40 mg L-1 and 60 mg L-1) sprayed at full bloom for both cultivars. An additional treatment of AVG 60 mg L-1 was sprayed one week after full bloom in 'Hosui'. The fruit set, number of fruit per tree, yield, fruit weight, seed number, and fruit quality attributes were assessed. Fruit set and yield of both cultivars are consistently increased by TDZ, within the rates of 20 to 60 mg L-1. Besides, its application increased fruit size of 'Hosui' and did not negatively affect fruit quality attributes of both cultivars.

The Role of Hydrogen Sulfide in Acute Liver Injury Induced by Traumatic Stress in Rats / 法医学杂志

OBJECTIVE@#To explore the role of hydrogen sulfide (H2S) in acute liver injury induced by crushing hind limbs of rats.@*METHODS@#The rats were randomly divided into the following groups: control, crushing, H2S donor sodium hydrosulfide (NaHS) + crushing, H2S inhibitor propargylglycine (PAG) + crushing group. The acute liver injury model was established by 'crushing the hind limbs of rats with standard weight. Rats were sacrificed at 30 min and 120 min after the crush. The activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured by colorimetric method, and the content of H2S in plasma and the contents of malondialdehyde (MDA), protein carbonyl, glutathione (GSH) in the liver and the activity of H2S generating enzyme (cystathionine y-lyase, CSE) were determined by chemical method. The expression of CSE mRNA in liver was detected by RT-PCR.@*RESULTS@#For crush injury group, the levels of AST and ALT in serum, MDA and protein carbonyl in liver increased. The levels of GSH, CSE, CSE mRNA in liver and H2S in serum decreased. The administration of NaHS before limbs crush could attenuate the changes of liver injury, but the pre-treatment with PAG could exacerbate the changes.@*CONCLUSION@#The decrease of H2S production could involve in mediating the acute liver injury induced by traumatic stress in rats.

Activation of group II metabotropic glutamate receptors blocks zinc release from hippocampal mossy fibers

BACKGROUND: The hippocampal CA3 area contains large amounts of vesicular zinc in the mossy fiber terminals which is released during synaptic activity, depending on presynaptic calcium. Another characteristic of these synapses is the presynaptic localization of high concentrations of group II metabotropic glutamate receptors, specifically activated by DCG-IV. Previous work has shown that DCG-IV affects only mossy fiber-evoked responses but not the signals from associational-commissural afferents, blocking mossy fiber synaptic transmission. Since zinc is released from mossy fibers even for single stimuli and it is generally assumed to be co-released with glutamate, the aim of the work was to investigate the effect of DCG-IV on mossy fiber zinc signals. RESULTS: Studies were performed using the membrane-permeant fluorescent zinc probe TSQ, and indicate that DCG-IV almost completely abolishes mossy fiber zinc changes as it does with synaptic transmission. CONCLUSIONS: Zinc signaling is regulated by the activation of type II metabotropic receptors, as it has been previously shown for glutamate, further supporting the corelease of glutamate and zinc from mossy fibers.

Post-treatment with plant extracts used in Brazilian folk medicine caused a partial reversal of the antiproliferative effect of glyphosate in the Allium cepa test

Species of the genus Psychotria are used for multiple purposes in Brazilian folk medicine, either as water infusions, baths or poultices. This study was aimed to evaluate the genotoxic and antiproliferative effects of infusions of Psychotria brachypoda and P. birotula on the Allium cepa test. Exposure to distilled water was used as a negative control, while exposure to glyphosate was used as a positive control. The interaction of extracts (as a post-treatment) with the effects of glyphosate was also studied. Results showed that glyphosate and the extracts of both P. brachypoda and P. birotula reduced the mitotic index as compared with the negative control (distilled water). Surprisingly, however, both extracts from P. brachypoda and P. birotula caused a partial reversal of the antiproliferative effect of glyphosat e when used as a post-treatment. Glyphosate also induced the highest number of cells with chromosomal alterations, which was followed by that of P. birotula extracts. However, the extracts from P. brachypoda did not show any significant genotoxic effect. Post-treatment of glyphosate-treated samples with distilled water allowed a partial recovery of the genotoxic effect of glyphosate, and some of the Psychotria extracts also did so. Notably, post-treatment of glyphosate-treated samples with P. brachypoda extracts induced a statistically significant apoptotic effect. It is concluded that P. brachypoda extracts show antiproliferative effects and are not genotoxic, while extracts of P. birotula show a less potent antiproliferative effect and may induce chromosomal abnormalities. The finding of a partial reversion of the effects of glyphosate by a post-treatment with extracts from both plants should be followed up.

Effect of endogenous hydrogen sulfide inhibition on structural and functional renal disturbances induced by gentamicin

Animal models of gentamicin nephrotoxicity present acute tubular necrosis associated with inflammation, which can contribute to intensify the renal damage. Hydrogen sulfide (H2S) is a signaling molecule involved in inflammation. We evaluated the effect of DL-propargylglycine (PAG), an inhibitor of endogenous H2S formation, on the renal damage induced by gentamicin. Male Wistar rats (N = 8) were injected with 40 mg/kg gentamicin (im) twice a day for 9 days, some of them also received PAG (N = 8, 10 mg·kg-1·day-1, ip). Control rats (N = 6) were treated with saline or PAG only (N = 4). Twenty-four-hour urine samples were collected one day after the end of these treatments, blood samples were collected, the animals were sacrificed, and the kidneys were removed for quantification of H2S formation and histological and immunohistochemical studies. Gentamicin-treated rats presented higher sodium and potassium fractional excretion, increased plasma creatinine [4.06 (3.00; 5.87) mg percent] and urea levels, a greater number of macrophages/monocytes, and a higher score for tubular interstitial lesions [3.50 (3.00; 4.00)] in the renal cortex. These changes were associated with increased H2S formation in the kidneys from gentamicin-treated rats (230.60 ± 38.62 µg·mg protein-1·h-1) compared to control (21.12 ± 1.63) and PAG (11.44 ± 3.08). Treatment with PAG reduced this increase (171.60 ± 18.34), the disturbances in plasma creatinine levels [2.20 (1.92; 4.60) mg percent], macrophage infiltration, and score for tubular interstitial lesions [2.00 (2.00; 3.00)]. However, PAG did not interfere with the increase in fractional sodium excretion provoked by gentamicin. The protective effect of PAG on gentamicin nephrotoxicity was related, at least in part, to decreased H2S formation.

Intellectual property rights related to the genetically modified glyphosate tolerant soybeans in Brazil

The present work analyzes the different modalities of protection of the intellectual creations in the biotechnology agricultural field. Regarding the Brazilian legislations related to the theme (the Industrial Property Law - no. 9. 279/96 and the Plant Variety Protection Law - no. 9. 456/97), and based in the international treaties signed by Brazil, the present work points to the inclusions of each of them, as well as to their interfaces using as reference the case study of glyphosate tolerant genetically modified soybean. For this case study, Monsanto's pipelines patents were searched and used to analyze the limits of patent protection in respect to others related to the Intellectual Property (IP) laws. Thus, it was possible to elucidate the complex scenario of the Intellectual Property of the glyphosate tolerant soybeans, since for the farmer it is hard to correlate the royalties payment with the IP enterprise's rights.

O presente trabalho analisa as diferentes modalidades de proteção das criações intelectuais no campo da biotecnologia agrícola. A partir das leis Brasileiras relacionadas ao tema (Lei da Propriedade Industrial - nº 9.279/96 e Lei da Proteção de Cultivares - nº 9.456/97), e com base nos tratados internacionais assinados pelo Brasil, o presente trabalho aponta as inclusões de cada uma, assim como, suas interfaces usando como referência o estudo de caso da soja geneticamente modificada para tolerância ao glifosato. Para este caso, patentes pipelines da Monsanto foram buscadas e usadas para analisar os limites de proteção das patentes frente às outras leis de Propriedade Intelectual (PI) relacionadas. Assim, foi possível elucidar o cenário complexo da Propriedade Intelectual das sojas tolerantes ao glifosato, já que para o agricultor não é fácil correlacionar o pagamento dos royalties com os direitos de PI da empresa.

5-HT1A autoreceptor modulation of locomotor activity induced by nitric oxide in the rat dorsal raphe nucleus

The dorsal raphe nucleus (DRN) is the origin of ascending serotonergic projections and is considered to be an important component of the brain circuit that mediates anxiety- and depression-related behaviors. A large fraction of DRN serotonin-positive neurons contain nitric oxide (NO). Disruption of NO-mediated neurotransmission in the DRN by NO synthase inhibitors produces anxiolytic- and antidepressant-like effects in rats and also induces nonspecific interference with locomotor activity. We investigated the involvement of the 5-HT1A autoreceptor in the locomotor effects induced by NO in the DRN of male Wistar rats (280-310 g, N = 9-10 per group). The NO donor 3-morpholinosylnomine hydrochloride (SIN-1, 150, and 300 nmol) and the NO scavenger S-3-carboxy-4-hydroxyphenylglycine (carboxy-PTIO, 0.1-3.0 nmol) were injected into the DRN of rats immediately before they were exposed to the open field for 10 min. To evaluate the involvement of the 5-HT1A receptor and the N-methyl-D-aspartate (NMDA) glutamate receptor in the locomotor effects of NO, animals were pretreated with the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT, 8 nmol), the 5-HT1A receptor antagonist N-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-N-2-pyridinyl-cyclohexanecarboxamide maleate (WAY-100635, 0.37 nmol), and the NMDA receptor antagonist DL-2-amino-7-phosphonoheptanoic acid (AP7, 1 nmol), followed by microinjection of SIN-1 into the DRN. SIN-1 increased the distance traveled (mean ± SEM) in the open-field test (4431 ± 306.1 cm; F7,63 = 2.44, P = 0.028) and this effect was blocked by previous 8-OH-DPAT (2885 ± 490.4 cm) or AP7 (3335 ± 283.5 cm) administration (P < 0.05, Duncan test). These results indicate that 5-HT1A receptor activation and/or facilitation of glutamate neurotransmission can modulate the locomotor effects induced by NO in the DRN.

The effect of the glyphosate, 2,4-D, atrazine e nicosulfuron herbicides upon the Edaphic collembola (Arthropoda: Ellipura) in a no tillage system

The use of herbicides is a common and intensive practice in no tillage systems. The herbicides can influence, directly or indirectly, the population of edaphic arthropods. Collembola is a group that functions as a bio-indicator of soil conditions. The degree of abundance and diversity of Collembola provides the level of soil disturbance provoked by agricultural practices. This experiment was designed to compare the influence of herbicides on the population fluctuation of Collembola in a no-till soil preparation system. The work was conducted in a non irrigated no-till area at the Núcleo Experimental de Ciências Agrárias of the Universidade Federal de Mato Grosso do Sul (UFMS), Campus de Dourados, in soil planted with corn as a surface covering, during the period of December, 2002 to December, 2003. The data were analyzed according to a completely randomized model, in a split plot design. The plots received four types of herbicides: glyphosate, atrazine, 2,4-D and nicosulfuron. A fifth plot did not receive any herbicide (control), for a total of five treatment types. The sub plots were represented by their collection times (10, 20, 30 and 40 days after the herbicide applications). Both the type of herbicide and the time of data sampling influenced the Collembola population fluctuaction. The treatments with atrazine and 2,4-D caused the most reduction of the population of Collembola, depending on the time of application.

No plantio direto o uso de herbicidas é uma prática comum e intensiva, que influencia direta ou indiretamente a população de artrópodes da mesofauna edáfica. O grau de abundância e diversidade de Collembola comumente é indicado para comprovar a extensão de distúrbios de várias práticas agrícolas, pois esse grupo serve como bioindicador das condições do solo. Esta pesquisa teve como objetivo comparar a influência de alguns herbicidas na flutuação populacional de Collembola, em solo sob o sistema de plantio direto. O trabalho foi realizado em uma área de plantio direto de sequeiro, do Núcleo Experimental de Ciências Agrárias da Universidade Federal de Mato Grosso do Sul (UFMS), Campus de Dourados, em latossolo roxo distroférrico com cobertura de milho, durante os meses de outubro de 2002 a janeiro de 2003. Os dados obtidos foram analisados segundo o modelo inteiramente casualizado constituído de tratamentos dispostos no esquema de parcelas subdivididas, onde as parcelas são representadas por uma testemunha mais quatro herbicidas: glifosate, atrazina, 2,4-D e nicosulfuron (totalizando cinco tratamentos) e as subparcelas pelas épocas de cada coleta (10, 20, 30 e 40 dias após aplicação dos herbicidas). Tanto os herbicidas testados quanto as épocas de coleta influenciaram a população de Collembola. Dependente do período de degradação dos herbicidas no solo, os tratamentos com 2,4-D, Atrazina, foram os que mais influenciaram a abundância de Collembola reduzindo sua população.

Glycine reduces tissue lipid peroxidation in hypoxia-reoxygenation-induced necrotizing enterocolitis in rats

OBJETIVO: Avaliar o efeito protetor da glicina, num modelo experimental de ECN. MÉTODOS: Foram utilizados 50 ratos Wistar recém-nascidos, com peso variando de 4 a 6 gramas, provenientes da ninhada de seis ratas. Cinco animais foram canibalizados e, os 45 restantes, foram distribuídos em três grupos: controle G1(n=12); G2(n=16), animais que foram submetidos à hipóxia-reoxigenação; G3(n=17), animais submetidos à hipóxia-reoxigenação após uma infusão intraperitoneal de glicina 5%. Os animais foram submetidos à hipóxia em uma câmara de CO2 recebendo um fluxo de ar contendo 100% de CO2, durante 5 minutos e à reoxigenação recebendo um fluxo de O2 a 100% por 5 minutos. Segmentos de intestino delgado e cólon de 1 cm de extensão foram preparados para análise histológica. O restante do intestino foi removido em bloco e congelado a menos 80°C para homogeneização e dosagem de malondialdeído tecidual (MDA). Classificou-se as lesões teciduais de Grau 0 a Grau 5, de acordo com a extensão da lesão mucosa. RESULTADOS: Os animais do Grupo G1 apresentaram graus de lesão de intestino delgado e cólon significantemente menores do que os animais dos Grupos G2 e G3. O grupo G2 apresentou valores médios de MDA significantemente maiores do que os animais do grupo G1 (p = .015) e G3 (p=0.021). Os animais dos grupos G1 e G3 apresentaram valores de MDA que não diferiram de forma significante (p = 0.992). CONCLUSÃO: A glicina diminuiu os níveis de MDA intestinais (um marcador da peroxidação lipídica) em ratos neonatais submetidos à hipóxia-reoxigenação.

Effect of ischaemia & aglycaemia on the synaptic transmission in neonatal rat spinal cord in vitro.

BACKGROUND & OBJECTIVES: In vitro models of anoxia have revealed severe changes in neuronal functions after ischaemia but not after aglycaemia, although hypoglycaemia produced severe neuronal dysfunctions sometimes leading to coma. The present study was therefore undertaken to examine and compare the effects of aglycaemia with that of ischaemia on synaptic transmission in vitro. METHODS: Spinal cord from the neonatal rat was isolated, hemisected and placed in a chamber perfused with standard physiological solution. The stimulation of a dorsal root elicited monosynaptic (MSR) and polysynaptic (PSR) reflex potentials in the segmental ventral root. The effects of suprefusing glucose free medium (aglycaemia) and superfusing glucose free and O2 free medium (ischaemia) were examined on these reflexes. RESULTS: Superfusion of aglycaemic solution did not alter the magnitude of MSR or PSR in the first 15 min and subsequently there was a time-dependent depression of the reflexes (P < 0.05). The ischaemic solution depressed the reflexes in a time-dependent manner from the very beginning. The 50 per cent depression of the reflexes occurred around 25 and 15 min, for aglycaemia and ischaemia, respectively. In the presence of Mg2+, the aglycaemia-induced depression of MSR was completely blocked but the ischaemic response was attenuated partially as the reflex was abolished by 80 min. INTERPRETATION & CONCLUSION: The results of the present study indicate that the aglycaemia and ischaemia depressed the synaptic transmission to the same extent though there were differences in their onset and progress. Aglycaemia involves N-methyl-D-aspartate (NMDA) receptor-dependent (Mg2+ sensitive) mechanism, while ischaemia-induced depression involves other mechanisms in addition to NMDA.

Effect of oxygen derived free radicals and glycine on sodium-potassium adenosine triphosphatase in the basolateral membrane of the kidney in ischemia-reperfusion

The aim of the present study was to examine the effect of exposing rats to ischemia-reperfusion while breathing 100% oxygen or room air, to find the effect of glycine on renal sodium-potassium adenosine triphosphatase [Na +/- K+ATPase] and endogenous antioxidant enzymes, superoxide dismutase and catalase, also to ascertain the effect of ischemia-reperfusion on renal nitric oxide and lipid peroxides. This study was carried out at King Saud University, Riyadh, Kingdom of Saudi Arabia, over a period of 11 months, February to December 2001. All previous measurements were carried out on the renal homogenate after 60 minutes ischemia, then after reperfusion while animals breathed room air or 100% oxygen and also after glycine treatment. The activity of Na +/- K+ATPase, catalase and superoxide dismutase concentration was decreased significantly in the ischemic rats compared to the control, a further decrease was found after 20 minutes of reflow while breathing room air. Breathing 100% oxygen resulted in a significant decrease in catalase and Na +/- K+ATPase activity and concentration of superoxide dismutase, glycine caused insignificant change of these enzymes after ischemia-index of lipid peroxidation and nitric oxide they were significantly elevated following reperfusion while rats breathed room air and further elevation was noticed after breathing 100% oxygen. However, potassium and creatinine did not change in all study groups, showed significant decrease after ischemia and ischemia-reperfusion may be due to marked Na+ loss in urine and lack of Na+ reabsorption. The inhibition of superoxide dismutase and catalase can be explained by increased reactive oxygen species during reperfusion and hyperoxia, also due to nitric oxide production and lipid peroxidation as shown by high malondialdehyde. Lack of Na+K+ATPase can be contributed to loss of antioxidant enzymes, nitric oxide production, and high reactive oxygen species. Hyperoxia in ischemia-reperfusion induces severe damage to cellular defence mechanisms and enhances reactive oxygen species injury. Glycine, as antioxidant, is involved in kidney protection from massive injury induced by ischemia-reperfusion, protects renal antioxidant enzymes and Na +/- K+ATPase, normalizes malondialdehyde, and nitric oxide levels. This data further supports the possible role of glycine therapy as an adjunct in the treatment of renal failure

Limited proteolysis by trypsin influences activity of maize phosphoenolpyruvate carboxylase.

Maize phosphoenolpyruvate carboxylase (PEPC) was rapidly and completely inactivated by very low concentrations of trypsin at 37 degrees C. PEP+Mg2+ and several other effectors of PEP carboxylase offered substantial protection against trypsin inactivation. Inactivation resulted from a fairly specific cleavage of 20 kDa peptide from the enzyme subunit. Limited proteolysis under catalytic condition (in presence of PEP, Mg2+ and HCO3) although yielded a truncated subunit of 90 kDa, did not affect the catalytic function appreciably but desensitized the enzyme to the effectors like glucose-6-phosphate glycine and malate. However, under non-catalytic condition, only malate sensitivity was appreciably affected. Significant protection of the enzyme activity against trypsin during catalytic phase could be either due to a conformational change induced on substrate binding. Several lines of evidence indicate that the inactivation caused by a cleavage at a highly conserved C-terminal end of the subunit.

Factors affecting transformation efficiency of BCG with a Mycobacterium-Escherichia coli shuttle vector pYUB18 by electroporation

BCG has been one of the vehicles for multi-recombinant vaccine. However, low transformation efficiency of BCG with plasmid DNA hampered studies involving expression of foreign antigens in BCG. In an effort to determine the optimal conditions, this study was initiated to investigate factors involved in the transformation of BCG with a Mycobacterium-Escherichia coli shuttle vector, pYUB18, by electroporation. Mycobacterium bovis BCG (strain 1173P2) was grown in Middlebrook (M) 7H9 broth containing albumin-dextrose-catalase and 0.05% tween 80, and transformed BCG was grown in M7H10 agar containing kanamycin for counting viable cells. Pretreatment of BCG with 10 mM CaCl2 improved the transformation efficiency, but overnight incubation of BCG with 1% glycine did not. The transformation efficiency in BCG also varied depending on voltage, resistance, and DNA concentration. The maximum transformation efficiency was obtained when the infinity resistance, 12.5 Kv/cm, and 100 ng of DNA were used, and reached 1.4 x 10(5) CFU/microgram of plasmid DNA, which is about 3-100 times greater than those from previous reports. The transformation conditions described in this study, therefore, will give us a better position for employing BCG as a vehicle for developing multi-recombinant vaccines.

Autonomic processing of the cardiovascular reflexes in the nucleus tractus solitarii

The nucleus tractus solitarii (NTS) receives afferent projections from the arterial baroreceptors, carotid chemoreceptors and cardiopulmonary receptors and as a function of this information produces autonomic adjustments in order to maintain arterial blood pressure within a narrow range of variation.The activation of each of these cardiovascular afferents produces a specific autonomic response by the excitation of neuronal projections from the NTS to the ventrolateral areas of the medulla (nucleus ambiguus, caudal and rostral ventrolateral medulla). The neurotransmitters at the NTS level as well as the excitatory amino acid (EAA) receptors involved in the processing of the autonomic responses in the NTS, although extensively studied, remain to be completely elucidated. In the present review we discuss the role of the EAA L-glutamate and its different receptor subtypes in the processing of the cardiovascular reflexes in the NTS. The data presented in this review related to the neurotransmission in the NTS are based on experimental evidence obtained in our laboratory in unanesthetized rats. The two major conclusions of the present review are that a) the excitation of the cardiovagal component by cardiovascular relfex activation (chemo- and Bezold-Jarisch reflexes) or by L-glutamatae microinjection into the NTS is mediated by N-methyl-D-aspartate (NMDA) receptors, and b) the sympatho-excitatory componente of the chemoreflex and the pressor response to L-glutamate microinjected into the NTS are not affected by an NMDA receptor antagonist, suggesting that the sympatho-excitatory component of these responses is mediated by non-NMDA receptors.

Clycine blocks the pressor response to L-glutamate microinjected into the nucleus tractus solitarii of conscious rats

Microinjection of L-glutamate into the nucleous tractus solitarii (NTS) of conscious freely moving Wistar rats (240-260 g) produces pressor (+48 ñ 4mmHg) and bradicardic (-153 ñ 20 bpm) responses. In the present study L-glutamate (2.5 nmol/100 nl) was microinjected before and after microinjection of increasing doses of glycine (10, 25 and 50 nmol/100 nl, N = 6) or saline (vehicle/100nl, N = 6) into the NTS. Microinjections of increasing doses of glycine into the NTS produced a dose-dependent reduction in the pressor but not in the bradycardic responses to L-glutamate. [10 nmol (+29 ñ 5mmHg and -110 ñ 18 bpm), 15 nmol (+12 ñ 7 mmHg and -88 ñ 21 bpm) and 50 nmol (+4 ñ 2 mmHg and -100 ñ 31 bpm)] The dose-dependent blockade of the pressor response to L-glutamate by glycine suggests an inhibitory neuromodulatory role for this amino acid in the sympatho-excitatory activity produced by L-glutamate microinjection into the NTS

Cardiorespiratory alterations produced by pharmacological agents applied to the ventrolateral medulla (intermediate area) of the cat: effect of exercise condition on glycine-induced inhibition

1. To study the action of the intermediate area (IA), coextensive with the rostral ventrolateral medulla, on the neurophysiological mechanisms involved in the regulation of respiration, in terms of inspiratory drive and respiratory timing, cats were submitted to topical application of sodium pentobarbital (30 mg/ml), leptazol (200 mg/ml), glutamate (50 mg/ml) and glycine (100 and 50 mg/ml) to the IA. The effects of electrically induced exercise on the ventilatory response and oxygen uptake (VO2) obtained by topical application of glycine (50 mg/ml) to the IA were also studied. 2. Leptazol reduced minute ventilation (VE) and inspiratory drive (VT/TI) and changed the timing mechanism. Glutamate only increased tidal volume (VT), VE and VT/TI. Arterial blood pressure (AP) increased and heart rate (HR) did not change with either drug. 3. Sodium pentobarbital reduced VT and changed the timing mechanism. Glycine only reduced VE, VT and VT/TI. AP decreased and HR did not change with either drug. 4. The depressor effects of glycine on respiratory pattern, VO2 and CO2 production (VCO2) tended to be attenuated by exercise. 5. The fall in AP due to glycine application did not differ between resting and exercise conditions. 6. Our results indicate that at least two different nervous structures are involved in the IA: one responsible for the respiratory drive and sensitive to glycine and glutamate, and the other responsible for the regulation of the timing mechanism and sensitive to sodium pentobarbital and leptazol