Induced corneal ulcers in cats - effects of 2% dorzolamide on epithelization time and on the expression of matrix metalloproteinase-9

Background: Topically administered 2% dorzolamide is among the most commonly used agents to lower IOP. As a complication of glaucoma, blind patients may develop corneal ulcers secondary to trauma. Nonetheless, in patients with a hypertensive or glaucomatous eye, in which the cornea has also been ulcerated, medical hypotensive therapy should not be discontinued. Therefore, the present study aimed to determine whether the instillation of a benzalkonium chloride (BAK)-preserved 2% dorzolamide alters corneal wound healing time and the levels of matrix metalloproteinases (MMP-9) in the tears of cats with experimentally induced corneal ulcers. Materials, Methods & Results: Sixteen cats (8/group) were randomly assigned to receive 40 µL of 2% dorzolamide (TG) or saline (CG) 3 times daily until corneal re-epithelialization. Experimental keratectomies were performed under general and topical anesthesia using an operating microscope. For this purpose, a millimitred trephine was calibrated and used to create a temporal paraxial corneal ulcer with a diameter of 6 mm and a depth of 200 µm. After corneal wounding, the ulcerated area, the healing time, blepharospasm, conjunctival hyperemia, and aqueous flare were compared between groups. Tears were collected at baseline and 24 and 48 h after keratectomy, and the total MMP-9 was quantified by ELISA. Data were assessed statistically using unpaired Student's t test, one-way, and two-way ANOVA followed by a Bonferroni post hoc test. Statistical significance was set at P < 0.05 for all analyses. The average time to achieve corneal wound healing did not differ between groups (P = 0.36) and was 65.50 ± 3.62 h in the CG and 71.00 ± 4.58 h in the TG. Twenty-four h after keratectomy, the ulcerated area in the CG was 3.34 mm2 larger than that observed in the TG (P = 0.04); the rest of the comparisons did not reach statistical significance at any time point between groups (P > 0.05). Higher blepharospasm scores were observed in cats of TG (P = 0.04). When compared with baseline of both groups, the levels of MMP-9 increased significantly at 24 and 48 h post-keratectomy (P < 0.001), but differences between groups were not observed at 24 and 48 h post-keratectomy (P > 0.05). Discussion: In cats, 9 mm axial corneal ulcers created by superficial debridement re-epithelize approximately 48 h postwounding. In the present study, re-epithelialization post keratectomy occurred within an average time of 68.25 h in most cats and in a delayed manner in one cat of the TG after 96 h. In the current study, the lesions in both groups healed without corneal scarring, pigmentation, or vascularization. Although BAC was present in all topical medications used in the present study, the authors attribute the higher scores of blepharospasm in the TG to the rheological characteristics and the pH of the dorzolamide ophthalmic solution. Indeed, the pH value of dorzolamide (5.58) may cause signs of irritation, as the tear film has an approximate pH of 7.6. Previous studies showed that ulcerated corneas presented significantly higher levels of MMP-9 in tears at the early stages (8 to 36 h) post-wounding. In the current study, the levels of this enzyme after wounding did not change significantly in the tears of cats treated with 2% dorzolamide when compared to the eyes in the control group. This study showed that the instillation of a BAC-preserved 2% dorzolamide ophthalmic solution did not impair the corneal wound healing time or the early expression of MMP-9 in the tears of cats with experimentally induced corneal ulcers. However, our results warrant further investigation in patients with ocular hypertension or glaucoma presenting concomitant naturally occurring corneal ulcers to certify our findings.

Dynamics of ammonia volatilization from NBPT-treated urea in tropical acid soils

The urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) reduces NH3 losses from urea (UR) surface-applied to soils, but its efficacy may be lower in acidic soils. The period when urease inhibition occurs efficaciously may change with soil pH. This needs to be clarified in tropical soils which are commonly acidic. This study evaluated the effectiveness of NBPT-treated urea to delay and reduce ammonia volatilization in two soils at three pH levels. Two experiments were conducted under laboratory conditions in soils with different textures (sandy-clay and clay). The treatments consisted of three soil pH levels and two N sources (UR and UR + NBPT), with five replicates. The soil pH values were adjusted and reached values of 4.5, 5.6, and 6.4 in the sandy-clay, and 4.5, 5.4, and 6.1 in the clay soil. Ammonia volatilization was measured using glass chambers (1.5 L). In the sandy-clay soil, NH3 losses were 40-47 % of the UR-N. In the clay soil, losses were 26-32 %. The addition of NBPT to UR reduced the NH3 volatilization by 18-53 %; the inhibitor decreased the N losses under all soil pH conditions but was significantly less efficient in acidic soils (pH 4.5). The lower efficiency of the inhibitor under acidic conditions was more evident in the first few days: 50 % of the total NH3 losses occurred in less than four days in soils with pH 4.5, but in 8-11 days in soils with pH above 5.4. The rapid loss in efficiency in more acidic soils is a drawback. Using NBPT in severely acidic soils showed a relatively small advantage over untreated UR as the inhibitor did not provide extra time for fertilizer incorporation and further reduction of NH3 losses.(AU)

Total phenolic and flavonoid contents, antioxidant, and α-glucosidase inhibitory activities of several big chili (Capsicum annuum L.) genotypes / Conteúdo fenólico total e flavonóide, atividades antioxidantes e inibitórias da α-glucosidase de vários genótipos de pimentão (Capsicum annuum L.)

ABSTRACT: Big chili has the largest fruit size compared to other types of chili. In addition to having a spicy taste, big chilies contain other beneficial biochemicals such as phenolics, flavonoids, antioxidants, and α-glucosidase inhibitors that have the potential to be developed for functional tropical plants. Information on the content is still not widely found. Therefore, this study identified biochemical compounds (TPC, TFC, DPPH, FRAP, AGI) in several genotypes of big chili peppers. From October to November 2021, we conducted a study using the microplate reader method. We repeated the experiment three times. The research design used was a completely randomized design with a single factor genotype. A total of 22 genotypes of big chili were used in this study. The big chili genotype F6074136-2-3-2-3 had the highest content of α-glucosidase inhibitor and antioxidant FRAP compared to other genotypes 67.62% and 71.20 mol TE/g extract. The genotypes CK 12 and F6074077-1-1-3-1 indicated the highest phenolic and flavonoid contents. The correlation between biochemical parameters showed that TPC-AGI and TPC-DPPH were significantly negatively correlated. The conclusion is that the information in this study can be the basis for the development of functional tropical plants in future research.

RESUMO: O pimentão grande tem o maior tamanho de fruta em comparação com outros tipos de pimentão. Além de ter um sabor picante, os pimentões grandes contêm outros bioquímicos benéficos, como fenólicos, flavonóides, antioxidantes e inibidores de α-glicosidase, que têm potencial para serem desenvolvidos para plantas tropicais funcionais. Informações sobre o conteúdo ainda não são amplamente encontradas. Portanto, este estudo teve como objetivo identificar compostos bioquímicos (TPC, TFC, DPPH, FRAP, AGI) em diversos genótipos de pimentão. O estudo foi conduzido de outubro a novembro de 2021 usando o método do leitor de microplaca. Os dados obtidos foram repetidos três vezes. O desenho de pesquisa usado foi um desenho inteiramente casualizado com um genótipo de fator único. Um total de 22 genótipos de pimentão grande foi usado neste estudo. O genótipo big chili F6074136-2-3-2-3 apresentou o maior teor de inibidor de α-glucosidase e antioxidante FRAP em comparação com outros genótipos 67,62% e 71,20 mol TE / g de extrato. O maior teor de fenólicos e flavonóides neste estudo foi indicado pelos genótipos CK 12 e F6074077-1-1-3-1. A correlação entre os parâmetros bioquímicos mostrou que TPC-AGI e TPC-DPPH foram significativamente correlacionados negativamente. Conclui-se que as informações deste estudo podem servir de base para o desenvolvimento de plantas tropicais funcionais em pesquisas futuras.

Abscisic acid and in vitro conservation of Manihot wild species

Abscisic acid (ABA) is associated with bud dormancy, leaf abscission, and germplasm growth inhibition in in vitro conservation. We evaluated the effects of ABA in four wild Manihot accessions and one cassava accession (M. esculenta Crantz) to refine in vitro conservation methods for these species. The experiment was performed at the Laboratory for Tissue Culture from Embrapa, Cruz das Almas, Bahia State, Brazil. The statistical design was completely random in a 5 × 5 factorial scheme [(5 ABA dosages (0, 0.25, 0.50, 0.75, and 1 mg L-1) and 5 Manihot species (M. pseudoglaziovii, M. tristis, M. flabellifolia, M. chlorosticta, and M. esculenta)], with 15 replicates. Mini-cuttings of 1 cm were used, each inoculated in 10 mL of modified Murashige and Skoog medium, solidified with Phytagel® (2.4 g L-1) containing the respective ABA dosages. Tubes containing these mini-cuttings were placed in a germplasm conservation room with an irradiance of 30 µmol m-2 s-1, temperature of 22 ± 1°C, and photoperiod of 12 hours. Plant height (cm), the number of living and senescent leaves, shoots, and mini-cuttings (1 cm), and fresh and dry weights of shoots and roots (mg) were evaluated after 150 days. Growth reduction was prominent in M. pseudoglaziovii, M. tristis, and M. flabellifolia during the in vitro conservation period. In the present study, the addition of ABA did not promote the expected reduction in plant growth.(AU)

Perinatal exposure to an aromatase inhibitor glyphosate-base herbicide reduced male and female social behavior in juvenile age and the sexual behavior at adult female rats / Exposição perinatal ao inibidor da aromatase a base de glifosato reduz o comportamento social de machos e fêmeas e o comportamento sexual de ratas

Objectives: The herbicide glyphosate, a pesticide used in agriculture to control weeds, both in food crops and in other agricultural areas, has been identified as an endocrine modulator through the inhibition of aromatase activity and the activation of estrogen receptors. The present study examined the effects of a glyphosate-based herbicide (Roundup® (GLY-BH) on sexual dimorphism of rats after perinatal exposure to low and high GLY-BH in males and females offspring. Methods: Two groups of pregnant rats were treated with two doses of GLY-BH (50 or 150 mg/kg) from day 15 of gestation (GD15) to postnatal day 7 (PND7). Play fighting behavior was observed at the juvenile stage and during social and sexual behaviors in adulthood. Results: Perinatal GLY-BH exposure reduced male and female body weight at 28, 75, and 90 days of age. The play fighting behavior was decreased in both sexes, but female rats were more affected. The sexual behaviors were reduced only in females. Conclusions: Perinatal exposure to both doses of GLY-BH promoted sexually dimorphic effects in both juvenile and adulthood stages. These effects were attributed to the inhibition of aromatase activity induced by exposure to GLY-BH in the perinatal period.(AU)

Objetivos: O glifosato é um herbicida não seletivo, usado em muitas culturas alimentares e não alimentares e em áreas não agrícolas, sendo que os produtos a base de glifosato atuam como moduladores das funções endócrinas por meio da inibição da atividade da aromatase e da ativação de receptores de estrógeno. O presente estudo avaliou os efeitos do herbicida Roundup® (GLY-BH) à base de glifosato, em comportamentos sexualmente dimórficos de ratos após exposição perinatal a doses baixas e altas de GLY-BH no período perinatal. Métodos: Ratas prenhas foram tratadas com 50 ou 150 mg/kg de GLY-BH do 15º dia de gestação (GD15) ao 7º dia de lactação (LD7). O comportamento de luta/brincar foi observado na fase juvenil e os comportamentos social e sexual na idade adulta. Resultados: a exposição perinatal a GLY-BH reduziu o peso corporal de machos e fêmeas aos 28, 75 e 90 dias de idade. O comportamento de luta/brincar diminuiu em ambos os sexos, sendo as ratas foram as mais afetadas. O comportamento sexual foi reduzido apenas nas fêmeas. Conclusões: A exposição perinatal a ambas as doses do GLY- BH promoveu tanto na idade juvenil como na idade adulta, efeitos sexualmente dimórficos. Esses efeitos foram atribuídos à inibição da atividade da aromatase induzida exposição perinatal ao GLY-BH.(AU)

Evaluation of potential hepatotoxicity induced by Bortezomib

Bortezomib, an inhibitor of 26S proteasome, is an anti-cancer therapeutic agent used in different cancer types. It leads to the arrest of the cancerous cell cycle by inhibiting angiogenesis and inducing apoptosis. Liver is the vital organ for detoxification and excretion of toxic products. The treatment with chemotherapy is a challenge, drugs are used to destroy cancer cells, but healthy cells can be affected during cancer treatment as well. The main objective of this study was to analyze the histopathological and biochemical effects of bortezomib on liver. Twenty-four female C57BL/6 mice were distributed into 4 groups, bortezomib injected treatment groups (Btz1, Btz2) and saline injected control groups (C1, C2). Bortezomib and saline were treated twice per week for 6 weeks and sacrificed at the end of one day (Btz1, C1) and 4 weeks (Btz2, C2) after the last injection. Liver samples were examined for histopathological analysis and the serum samples processed for biochemical analysis. Tissue samples were fixed, routinely processed, sectioned, and stained with Hematoxylin and Eosin (H&E). Periodic Acid-Schiff (PAS), Sudan Black staining and Masson's trichrome histochemical staining methods were performed to characterize the lesions. Histopathological analysis of the Btz1 and Btz2 groups revealed acute hepatic morphological changes such as hepatocellular swelling (cloudy swelling), necro-inflammatory reaction, and increased mononuclear polyploidy. Based on the negative staining with PAS and Sudan Black staining, hepatocellular swelling was diagnosed as hydropic degeneration. Necro-inflammatory reaction observed in the form of acute hepatitis was composed of mainly mononuclear cell infiltration accompanied by multifocal necrotic foci. Kupffer cell proliferation was observed in parallel with degenerative and necrotic changes. An Increase in hepatocellular mononuclear polyploidy visualized as hepatocytes with a single enlarged nucleus was detected in all liver sections of Btz1 and Btz2 groups. Individual cases of cholestasis (n = 1) and mild hepatic fibrosis (n = 1) were also reported. Significant elevated levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) were detected in bortezomib treated groups. Few clinical cases reported liver injury related to bortezomib used for cancer treatment. However, the liver was not considered as a target for bortezomib treatment. Our data suggesting that bortezomib caused liver damage and induces elevations in serum levels. The reported hepatic lesions including hepatocellular swelling, acute hepatitis and mononuclear polyploidy were mainly mild and moderate in severity. The increase of polyploidy in liver tissue of mice treated with bortezomib in this study was explained as a reaction of the liver facing the drug-induced hepatic damage. The mechanism leading to the hepatotoxicity of bortezomib treatment is not known but the production of a toxic metabolite through its metabolism in the liver can be suggested. Moreover, no recovery was also observed in histopathological and biochemical analyses suggesting that the bortezomib effect is non-reversible four weeks after the drug was withdrawn. Patients should be informed about the possibility of acute drug-induced hepatitis and hepatotoxicity of this chemotherapeutic agent after the treatment.(AU)

Abscisic acid and in vitro conservation of Manihot wild species / Abscisic acid and in vitro conservation of Manihot wild species

Abscisic acid (ABA) is associated with bud dormancy, leaf abscission, and germplasm growth inhibition in in vitro conservation. We evaluated the effects of ABA in four wild Manihot accessions and one cassava accession (M. esculenta Crantz) to refine in vitro conservation methods for these species. The experiment was performed at the Laboratory for Tissue Culture from Embrapa, Cruz das Almas, Bahia State, Brazil. The statistical design was completely random in a 5 × 5 factorial scheme [(5 ABA dosages (0, 0.25, 0.50, 0.75, and 1 mg L-1) and 5 Manihot species (M. pseudoglaziovii, M. tristis, M. flabellifolia, M. chlorosticta, and M. esculenta)], with 15 replicates. Mini-cuttings of 1 cm were used, each inoculated in 10 mL of modified Murashige and Skoog medium, solidified with Phytagel® (2.4 g L-1) containing the respective ABA dosages. Tubes containing these mini-cuttings were placed in a germplasm conservation room with an irradiance of 30 µmol m-2 s-1, temperature of 22 ± 1°C, and photoperiod of 12 hours. Plant height (cm), the number of living and senescent leaves, shoots, and mini-cuttings (1 cm), and fresh and dry weights of shoots and roots (mg) were evaluated after 150 days. Growth reduction was prominent in M. pseudoglaziovii, M. tristis, and M. flabellifolia during the in vitro conservation period. In the present study, the addition of ABA did not promote the expected re

Abscisic acid (ABA) is associated with bud dormancy, leaf abscission, and germplasm growth inhibition in in vitro conservation. We evaluated the effects of ABA in four wild Manihot accessions and one cassava accession (M. esculenta Crantz) to refine in vitro conservation methods for these species. The experiment was performed at the Laboratory for Tissue Culture from Embrapa, Cruz das Almas, Bahia State, Brazil. The statistical design was completely random in a 5 × 5 factorial scheme [(5 ABA dosages (0, 0.25, 0.50, 0.75, and 1 mg L-1) and 5 Manihot species (M. pseudoglaziovii, M. tristis, M. flabellifolia, M. chlorosticta, and M. esculenta)], with 15 replicates. Mini-cuttings of 1 cm were used, each inoculated in 10 mL of modified Murashige and Skoog medium, solidified with Phytagel® (2.4 g L-1) containing the respective ABA dosages. Tubes containing these mini-cuttings were placed in a germplasm conservation room with an irradiance of 30 µmol m-2 s-1, temperature of 22 ± 1°C, and photoperiod of 12 hours. Plant height (cm), the number of living and senescent leaves, shoots, and mini-cuttings (1 cm), and fresh and dry weights of shoots and roots (mg) were evaluated after 150 days. Growth reduction was prominent in M. pseudoglaziovii, M. tristis, and M. flabellifolia during the in vitro conservation period. In the present study, the addition of ABA did not promote the expected re

Apoptosis (programmed cell death) and its signals: a review / Apoptose, morte celular programada e seus sinais: uma revisão

Apoptosis is a sequential order of cell death occurring regularly to ensure a homeostatic balance between the rate of cell formation and cell death. However, a misplaced of this balancing function can contribute to an abnormal cell growth / proliferation or autoimmune disorders etc. Apoptosis is therefore said to be crucial from the point of development of an embryo throughout the growth of an organism contributing to the renewal of tissues and also the getting rid of inflammatory cells. This review seeks to elaborate on the recent overview of the mechanism involved in apoptosis, some element and signal contributing to its function and inhibition together with how their malfunction contribute to a number of cancer related cases.(AU)

A apoptose é uma ordem sequencial de morte celular que ocorre regularmente para garantir um equilíbrio homeostático entre a taxa de formação e a morte celular. No entanto, um desequilíbrio dessa função pode contribuir para um crescimento/proliferação celular anormal ou distúrbios autoimunes. A apoptose é, portanto, considerada crucial do ponto de desenvolvimento de um embrião ao longo do crescimento de um organismo que contribui para a renovação dos tecidos e também a eliminação de células inflamatórias. Esta revisão procura elaborar a recente visão geral do mecanismo envolvido na apoptose, alguns elementos e sinais que contribuem para sua função e inibição, além de como o mau funcionamento deles contribui para vários casos relacionados ao câncer.(AU)

Performance of RR soybean submitted to postemergence application of glyphosate with a foliar elicitor product / Desempenho da soja RR submetida a aplicação de glifosato associado a produto elicitor foliar em pós-emergência

With the advent of glyphosate-resistant soybean, postemergence applications of the herbicide have become routine. In this way, problems related to the selectivity of glyphosate have been shown to be increasingly intense, due to the use of increased dosages and applications of the herbicide at unrecommended moments. In order to evaluate the agronomic performance of soybean plants subjected to the postemergence application of glyphosate ­ both in isolation and together with Crop+ ­ at different stages and dosage levels, a randomized block design experiment was carried out with two adjacent checks and four repetitions. The treatments consisted of: glyphosate (1296 g·ha-1); glyphosate + Crop+ (1296 + 0.25 g or L·ha-1); glyphosate + Crop+ (1296 + 0.5 g or L·ha-1); glyphosate (2592 g·ha-1); glyphosate + Crop+ (2592 + 0.25 g or L·ha-1); glyphosate + Crop+ (2592 + 0.5 g or L·ha-1); 2 × glyphosate (2 × 1296 g·ha-1); 2 × glyphosate + Crop+ (2 × 1296 + 0.25 g or L·ha-1); 2 × glyphosate + Crop+ (2 × 1296 + 0.5 g or L·ha-1) and Crop+ (0.5 g·ha-1). The phytotoxicity, soil plant analysis development (SPAD) index, height, stand, mass of 100 grains, and yield of the soybean plants were evaluated. The use of Crop+ on soybean plants led to an increase in the SPAD index; the use of Crop+ in association with glyphosate made possible the attenuation of visible symptoms of damage, preventing reductions in crop yield.(AU)

Com o advento da soja resistente ao glifosato, aplicações em pós-emergência da cultura tornaram-se corriqueiras. Dessa forma, problemas relacionados à seletividade desse herbicida mostraram-se cada vez mais intensos, devido ao aumento da dose utilizada e à aplicação em momentos não recomendados. Para avaliar o desempenho agronômico da soja submetida à aplicação de glifosato isolado em pós-emergência e em associação ao Crop+ em diferentes estádios e doses, um experimento foi realizado em delineamento de blocos ao acaso, em esquema de testemunhas duplas adjacentes, adotando-se quatro repetições. Os tratamentos foram constituídos por: glifosato (1296 g·ha-1); glifosato + Crop+ (1296 + 0,25 g ou L·ha-1); glifosato + Crop+ (1296 + 0,5 g ou L·ha-1);glifosato (2592 g·ha-1); glifosato + Crop+ (2592 + 0,25 g ou L·ha-1);glifosato + Crop+ (2592 + 0,5 g ou L·ha-1); 2 × glifosato (2 × 1296 g·ha-1); 2 × glifosato + Crop+ (2 × 1296 + 0,25 g ou L·ha-1); 2 × glifosato + Crop+ (2 × 1296 + 0,5 g ou L·ha-1) e Crop+ (0,5 g·ha-1). Foram avaliados fitointoxicação das plantas de soja, índice SPAD (soil plant analysis development), altura, estande, massa de cem grãos e produtividade. Nos resultados, a utilização de Crop+ em plantas de soja proporcionou o aumento no índice SPAD e, quando o Crop+ foi associado ao glifosato, possibilitou a atenuação dos sintomas visuais de injúrias, prevenindo reduções na produtividade da cultura.(AU)

A Kunitz-type peptide from Dendroaspis polylepis venom as a simultaneous inhibitor of serine and cysteine proteases

Proteases play an important role for the proper physiological functions of the most diverse organisms. When unregulated, they are associated with several pathologies. Therefore, proteases have become potential therapeutic targets regarding the search for inhibitors. Snake venoms are complex mixtures of molecules that can feature a variety of functions, including peptidase inhibition. Considering this, the present study reports the purification and characterization of a Kunitz-type peptide present in the Dendroaspis polylepis venom as a simultaneous inhibitor of elastase-1 and cathepsin L. Methods: The low molecular weight pool from D. polylepis venom was fractionated in reverse phase HPLC and all peaks were tested in fluorimetric assays. The selected fraction that presented inhibitory activity over both proteases was submitted to mass spectrometry analysis, and the obtained sequence was determined as a Kunitz-type serine protease inhibitor homolog dendrotoxin I. The molecular docking of the Kunitz peptide on the elastase was carried out in the program Z-DOCK, and the program RosettaDock was used to add hydrogens to the models, which were re-ranked using ZRANK program. Results: The fraction containing the Kunitz molecule presented similar inhibition of both elastase-1 and cathepsin L. This Kunitz-type peptide was characterized as an uncompetitive inhibitor for elastase-1, presenting an inhibition constant (Ki) of 8 μM. The docking analysis led us to synthesize two peptides: PEP1, which was substrate for both elastase-1 and cathepsin L, and PEP2, a 30-mer cyclic peptide, which showed to be a cathepsin L competitive inhibitor, with a Ki of 1.96 µM, and an elastase-1 substrate. Conclusion: This work describes a Kunitz-type peptide toxin presenting inhibitory potential over serine and cysteine proteases, and this could contribute to further understand the envenomation process by D. polylepis. In addition, the PEP2 inhibits the cathepsin L activity with a low inhibition constant.(AU)

Identification and characterization of the first endogenous phospholipase A2 inhibitor from a non-venomous tropical snake, Boa constrictor (Serpentes: Boidae)

Abstract Background: Endogenous phospholipase A2 inhibitors from snake blood (sbPLIs) have been isolated from several species around the world, with the primary function of self-protection against the action of toxic phospholipases A2. In American snakes, sbPLIs were solely described in pit vipers, in which the natural protection role is justified. In this study, we described a sbPLI in Boa constrictor (popularly known as jiboia), a non-venomous snake species from America. Methods: PLA2 inhibitory activity was tested in the blood plasma of B. constrictor using C. d. terrificus venom as the enzyme source. Antibodies developed against CNF, a sbγPLI from Crotalus durissus terrificus, were used to investigate the presence of homologues in the blood plasma of B. constrictor. A CNF-like molecule with a PLA2 inhibitory activity was purified by column chromatography. The encoding gene for the inhibitor was cloned from B. constrictor liver tissue. The DNA fragment was cloned, purified and sequenced. The deduced primary sequence of interest was aligned with known sbγPLIs from the literature. Results: The blood plasma of B. constrictor displayed PLA2 inhibitory activity. A CNF-like molecule (named BcNF) was identified and purified from the blood plasma of B. constrictor. Basic properties such as molecular mass, composing amino acids, and pI were comparable, but BcNF displayed reduced specific activity in PLA2 inhibition. BcNF showed highest identity scores (ISs) with sbγPLIs from pit vipers from Latin America (90-100%), followed by gamma inhibitors from Asian viperid (80-90%). ISs below 70% were obtained for BcNF and non-venomous species from Asia. Conclusion: A functional sbγPLI (BcNF) was described in the blood plasma of B. constrictor. BcNF displayed higher primary identity with sbγPLIs from Viperidae than to sbγPLIs from non-venomous species from Asia. The physiological role played by sbγPLIs in non-venomous snake species remains to be understood. Further investigation is needed.(AU)

Identification and characterization of the first endogenous phospholipase A2 inhibitor from a non-venomous tropical snake, Boa constrictor (Serpentes: Boidae)

Background:Endogenous phospholipase A2 inhibitors from snake blood (sbPLIs) have been isolated from several species around the world, with the primary function of self-protection against the action of toxic phospholipases A2. In American snakes, sbPLIs were solely described in pit vipers, in which the natural protection role is justified. In this study, we described a sbPLI in Boa constrictor (popularly known as jiboia), a non-venomous snake species from America.Methods:PLA2 inhibitory activity was tested in the blood plasma of B. constrictor using C. d. terrificus venom as the enzyme source. Antibodies developed against CNF, a sbγPLI from Crotalus durissus terrificus, were used to investigate the presence of homologues in the blood plasma of B. constrictor. A CNF-like molecule with a PLA2 inhibitory activity was purified by column chromatography. The encoding gene for the inhibitor was cloned from B. constrictor liver tissue. The DNA fragment was cloned, purified and sequenced. The deduced primary sequence of interest was aligned with known sbγPLIs from the literature.Results:The blood plasma of B. constrictor displayed PLA2 inhibitory activity. A CNF-like molecule (named BcNF) was identified and purified from the blood plasma of B. constrictor. Basic properties such as molecular mass, composing amino acids, and pI were comparable, but BcNF displayed reduced specific activity in PLA2 inhibition. BcNF showed highest identity scores (ISs) with sbγPLIs from pit vipers from Latin America (90-100%), followed by gamma inhibitors from Asian viperid (80-90%). ISs below 70% were obtained for BcNF and non-venomous species from Asia.Conclusion:A functional sbγPLI (BcNF) was described in the blood plasma of B. constrictor. BcNF displayed higher primary identity with sbγPLIs from Viperidae than...(AU)

A Kunitz-type peptide from Dendroaspis polylepis venom as a simultaneous inhibitor of serine and cysteine proteases

Proteases play an important role for the proper physiological functions of the most diverse organisms. When unregulated, they are associated with several pathologies. Therefore, proteases have become potential therapeutic targets regarding the search for inhibitors. Snake venoms are complex mixtures of molecules that can feature a variety of functions, including peptidase inhibition. Considering this, the present study reports the purification and characterization of a Kunitz-type peptide present in the Dendroaspis polylepis venom as a simultaneous inhibitor of elastase-1 and cathepsin L. Methods: The low molecular weight pool from D. polylepis venom was fractionated in reverse phase HPLC and all peaks were tested in fluorimetric assays. The selected fraction that presented inhibitory activity over both proteases was submitted to mass spectrometry analysis, and the obtained sequence was determined as a Kunitz-type serine protease inhibitor homolog dendrotoxin I. The molecular docking of the Kunitz peptide on the elastase was carried out in the program Z-DOCK, and the program RosettaDock was used to add hydrogens to the models, which were re-ranked using ZRANK program. Results: The fraction containing the Kunitz molecule presented similar inhibition of both elastase-1 and cathepsin L. This Kunitz-type peptide was characterized as an uncompetitive inhibitor for elastase-1, presenting an inhibition constant (Ki) of 8 μM. The docking analysis led us to synthesize two peptides: PEP1, which was substrate for both elastase-1 and cathepsin L, and PEP2, a 30-mer cyclic peptide, which showed to be a cathepsin L competitive inhibitor, with a Ki of 1.96 µM, and an elastase-1 substrate. Conclusion: This work describes a Kunitz-type peptide toxin presenting inhibitory potential over serine and cysteine proteases, and this could contribute to further understand the envenomation process by D. polylepis. In addition, the PEP2 inhibits the cathepsin L activity with a low inhibition constant.(AU)

Performance of RR soybean submitted to postemergence application of glyphosate with a foliar elicitor product / Desempenho da soja RR submetida a aplicação de glifosato associado a produto elicitor foliar em pós-emergência

With the advent of glyphosate-resistant soybean, postemergence applications of the herbicide have become routine. In this way, problems related to the selectivity of glyphosate have been shown to be increasingly intense, due to the use of increased dosages and applications of the herbicide at unrecommended moments. In order to evaluate the agronomic performance of soybean plants subjected to the postemergence application of glyphosate ­ both in isolation and together with Crop+ ­ at different stages and dosage levels, a randomized block design experiment was carried out with two adjacent checks and four repetitions. The treatments consisted of: glyphosate (1296 g·ha-1); glyphosate + Crop+ (1296 + 0.25 g or L·ha-1); glyphosate + Crop+ (1296 + 0.5 g or L·ha-1); glyphosate (2592 g·ha-1); glyphosate + Crop+ (2592 + 0.25 g or L·ha-1); glyphosate + Crop+ (2592 + 0.5 g or L·ha-1); 2 × glyphosate (2 × 1296 g·ha-1); 2 × glyphosate + Crop+ (2 × 1296 + 0.25 g or L·ha-1); 2 × glyphosate + Crop+ (2 × 1296 + 0.5 g or L·ha-1) and Crop+ (0.5 g·ha-1). The phytotoxicity, soil plant analysis development (SPAD) index, height, stand, mass of 100 grains, and yield of the soybean plants were evaluated. The use of Crop+ on soybean plants led to an increase in the SPAD index; the use of Crop+ in association with glyphosate made possible the attenuation of visible symptoms of damage, preventing reductions in crop yield.(AU)

Com o advento da soja resistente ao glifosato, aplicações em pós-emergência da cultura tornaram-se corriqueiras. Dessa forma, problemas relacionados à seletividade desse herbicida mostraram-se cada vez mais intensos, devido ao aumento da dose utilizada e à aplicação em momentos não recomendados. Para avaliar o desempenho agronômico da soja submetida à aplicação de glifosato isolado em pós-emergência e em associação ao Crop+ em diferentes estádios e doses, um experimento foi realizado em delineamento de blocos ao acaso, em esquema de testemunhas duplas adjacentes, adotando-se quatro repetições. Os tratamentos foram constituídos por: glifosato (1296 g·ha-1); glifosato + Crop+ (1296 + 0,25 g ou L·ha-1); glifosato + Crop+ (1296 + 0,5 g ou L·ha-1);glifosato (2592 g·ha-1); glifosato + Crop+ (2592 + 0,25 g ou L·ha-1);glifosato + Crop+ (2592 + 0,5 g ou L·ha-1); 2 × glifosato (2 × 1296 g·ha-1); 2 × glifosato + Crop+ (2 × 1296 + 0,25 g ou L·ha-1); 2 × glifosato + Crop+ (2 × 1296 + 0,5 g ou L·ha-1) e Crop+ (0,5 g·ha-1). Foram avaliados fitointoxicação das plantas de soja, índice SPAD (soil plant analysis development), altura, estande, massa de cem grãos e produtividade. Nos resultados, a utilização de Crop+ em plantas de soja proporcionou o aumento no índice SPAD e, quando o Crop+ foi associado ao glifosato, possibilitou a atenuação dos sintomas visuais de injúrias, prevenindo reduções na produtividade da cultura.(AU)

Metástase hepática de tumor gastrointestinal estromal (gist) em cão: relato de caso / Hepatic metastasis of gastrointestinal stromal tumors (gist) in dog: case report

Os tumores mesenquimais originados a partir de células intersticiais de Cajal, denominados tumores gastrointestinais estromais (GIST) são raros em humanos e com incidência ainda desconhecida em cães. A diferenciação de GIST, leiomiossarcoma e leiomioma com base apenas em exames de imagem, morfológicos e colorações simples por meio da histologia é muitas vezes difícil, sendo indicada realização de imuno-histoquímica. Foi atendida uma cadela, sem padrão racial definido, oito anos, 17kg de peso corporal, com discreto aumento de volume abdominal, sendo evidenciada por ultrassom abdominal neoformação intra-abdominal em região mesogástrica, sem demais alterações clínicas. Durante celiotomia exploratória, identificou-se que a neoformação intestinal localizava-se em jejuno, optando pela realização de enterectomia, sendo posteriormente diagnosticada como leiomiossarcoma pela análise histopatológica. Preconizou-se a instituição de quimioterapia adjuvante com o uso de doxorrubicina (30 mg/m²), a cada 21 dias, totalizando seis sessões. Após dez meses do tratamento cirúrgico, a paciente foi submetida a novo estadiamento clínico, sendo visibilizado, mediante ultrassom abdominal neoformações sólidas em fígado. A partir de biópsia hepática guiada por ultrassom com agulha “tru-cut” e análise histológica e imuno-histoquímica, definiu-se que as neoformações hepáticas eram metástases de GIST, instituindo o tratamento com fosfato de toceranib (Palladia®). Desde o momento da enterectomia ao óbito da paciente, contabilizou-se 20,4 meses de sobrevida global.

Mesenchymal tumors originating from interstitial cells of Cajal, called stromal gastrointestinal tumors (GIST) are rare in humans and their incidence is still unknown in dogs. The differentiation of GIST, leiomyosarcoma and leiomyoma based only on imaging, morphological and simple staining through histology is often difficult, and immunohistochemistry is indicated. A female dog, mixed breed, eight years old, 17kg of body weight, with a slight increase in abdominal volume, and ultrasound evidence of an intra-abdominal neoformation in the mesogastric region, without other clinical changes was seen. During exploratory celiotomy, it was identified that the intestinal neoformation in jejunum, opting for enterectomy, being identified as leiomyosarcoma according to histopathological analysis. Adjuvant chemotherapy was instituted using doxorubicin (30mg / m²) every 21 days, totaling six sessions. The patient underwent a new clinical staging, 10 months after enterectomy, where were visualized, by abdominal ultrasound, neoformations in the liver. Based on ultrasound guided liver biopsy with “tru-cut” needle and histological and immunohistochemical analysis, the neoformation was defined as GIST liver metastasis, being instituted treatment with toceranib phosphate (Palladia®). From the time of enterectomy to the death of the patient, 20,4 months of overall survival were counted.

Metástase hepática de tumor gastrointestinal estromal (gist) em cão: relato de caso / Hepatic metastasis of gastrointestinal stromal tumors (gist) in dog: case report

Os tumores mesenquimais originados a partir de células intersticiais de Cajal, denominados tumores gastrointestinais estromais (GIST) são raros em humanos e com incidência ainda desconhecida em cães. A diferenciação de GIST, leiomiossarcoma e leiomioma com base apenas em exames de imagem, morfológicos e colorações simples por meio da histologia é muitas vezes difícil, sendo indicada realização de imuno-histoquímica. Foi atendida uma cadela, sem padrão racial definido, oito anos, 17kg de peso corporal, com discreto aumento de volume abdominal, sendo evidenciada por ultrassom abdominal neoformação intra-abdominal em região mesogástrica, sem demais alterações clínicas. Durante celiotomia exploratória, identificou-se que a neoformação intestinal localizava-se em jejuno, optando pela realização de enterectomia, sendo posteriormente diagnosticada como leiomiossarcoma pela análise histopatológica. Preconizou-se a instituição de quimioterapia adjuvante com o uso de doxorrubicina (30 mg/m²), a cada 21 dias, totalizando seis sessões. Após dez meses do tratamento cirúrgico, a paciente foi submetida a novo estadiamento clínico, sendo visibilizado, mediante ultrassom abdominal neoformações sólidas em fígado. A partir de biópsia hepática guiada por ultrassom com agulha “tru-cut” e análise histológica e imuno-histoquímica, definiu-se que as neoformações hepáticas eram metástases de GIST, instituindo o tratamento com fosfato de toceranib (Palladia®). Desde o momento da enterectomia ao óbito da paciente, contabilizou-se 20,4 meses de sobrevida global.(AU)

Mesenchymal tumors originating from interstitial cells of Cajal, called stromal gastrointestinal tumors (GIST) are rare in humans and their incidence is still unknown in dogs. The differentiation of GIST, leiomyosarcoma and leiomyoma based only on imaging, morphological and simple staining through histology is often difficult, and immunohistochemistry is indicated. A female dog, mixed breed, eight years old, 17kg of body weight, with a slight increase in abdominal volume, and ultrasound evidence of an intra-abdominal neoformation in the mesogastric region, without other clinical changes was seen. During exploratory celiotomy, it was identified that the intestinal neoformation in jejunum, opting for enterectomy, being identified as leiomyosarcoma according to histopathological analysis. Adjuvant chemotherapy was instituted using doxorubicin (30mg / m²) every 21 days, totaling six sessions. The patient underwent a new clinical staging, 10 months after enterectomy, where were visualized, by abdominal ultrasound, neoformations in the liver. Based on ultrasound guided liver biopsy with “tru-cut” needle and histological and immunohistochemical analysis, the neoformation was defined as GIST liver metastasis, being instituted treatment with toceranib phosphate (Palladia®). From the time of enterectomy to the death of the patient, 20,4 months of overall survival were counted.(AU)

Apoptosis (programmed cell death) and its signals - A review

Abstract Apoptosis is a sequential order of cell death occurring regularly to ensure a homeostatic balance between the rate of cell formation and cell death. However, a misplaced of this balancing function can contribute to an abnormal cell growth / proliferation or autoimmune disorders etc. Apoptosis is therefore said to be crucial from the point of development of an embryo throughout the growth of an organism contributing to the renewal of tissues and also the getting rid of inflammatory cells. This review seeks to elaborate on the recent overview of the mechanism involved in apoptosis, some element and signal contributing to its function and inhibition together with how their malfunction contribute to a number of cancer related cases.

Resumo A apoptose é uma ordem sequencial de morte celular que ocorre regularmente para garantir um equilíbrio homeostático entre a taxa de formação e a morte celular. No entanto, um desequilíbrio dessa função pode contribuir para um crescimento/proliferação celular anormal ou distúrbios autoimunes. A apoptose é, portanto, considerada crucial do ponto de desenvolvimento de um embrião ao longo do crescimento de um organismo que contribui para a renovação dos tecidos e também a eliminação de células inflamatórias. Esta revisão procura elaborar a recente visão geral do mecanismo envolvido na apoptose, alguns elementos e sinais que contribuem para sua função e inibição, além de como o mau funcionamento deles contribui para vários casos relacionados ao câncer.

Efficacy of phosphocreatine pre-administration on XIAP and Smac in ischemic penumbra of rats with focal cerebral ischemia reperfusion injury

Purpose: To observe the efficacy of phosphocreatine pre-administration (PCr-PA) on X-linked inhibitor of apoptosis protein (XIAP), the second mitochondia-derived activator of caspase (Smac) and apoptosis in the ischemic penumbra of rats with focal cerebral ischemia-reperfusion injury (CIRI).Methods: A total of 60 healthy male Sprague Dawley (SD) rats were randomly divided into three groups (n=20): group A (the sham operation group), group B [intraperitoneally injected with 20 mg/kg (10 mg/ml) of saline before preparing the ischemia-reperfusion (IR) model], and group C [intraperitoneally injected with 20 mg/kg (10 mg/ml) of PCr immediately before preparing the IR model]. After 24 h for reperfusion, the neurological function was evaluated and the tissue was sampled to detect expression of XIAP, Smac and caspase-3 positive cells in the ischemic penumbra so as to observe the apoptosis.Results: Compared with group B, neurological deficit scores, numbers of apoptotic cells, expression of Smac,caspase-9 and the numbers of Caspase-3 positive cells were decreased while expression of XIAP were increased in the ischemic penumbra of group C.Conclusions: Phosphocreatine pre-administration may elicit neuroprotective effects in the brain by increasing expression of X-linked inhibitor of apoptosis protein, reducing expression of second mitochondia-derived activator of caspase, and inhibiting the apoptosis in the ischemic penumbra.(AU)

Concentration and periods of application of prohexadione calcium in the growth of strawberry seedlings / Concentrações e épocas de aplicação do proexadione cálcio no crescimento de mudas de morangueiro

For strawberry cultivation in Brazil, producers are dependent on imported seedlings. An alternative strategy to reduce this dependence is the use of seedlings obtained from nursery plants grown in a protected environment. However, as these seedlings are produced in the summer and planted at the end of this season or the spring of the following year, it is necessary to control growth to reduce the energy costs of the plants. The objective of this study was to evaluate the effect of different concentrations and periods of application of proexadione calcium (ProCa) on growth control of strawberry seedlings. The experiment was carried out in a greenhouse with seedlings of the cultivars "Aromas" and "Camarosa", produced by rooting stolons and kept in polystyrene trays of 72 cells in a substrate of carbonized rice husk. The experimental design was completely randomized, with a 4 × 2 factorial scheme (4 concentrations of ProCa: 0, 100, 200, and 400 mg L-¹ × 2 periods of application: at 20 and 30 days after the planting period of rooting stolon). Plant survival, crown diameter, petiole length, total leaf area, specific leaf area, chlorophyll concentration, and dry mass of the crown and shoot were evaluated. The application of ProCa at 20 days after the planting period of the rooting stolon at the concentrations of 200 and 400 mg L-¹ favored the reduction of petiole length in plants of "Aromas" strawberry and total leaf aerial in "Camarosa" strawberry. The application of ProCa from the concentration of 100 mg L-¹ reduced the vegetative growth of "Aromas" and "Camarosa" strawberry seedlings cultivated in substrate.(AU)

Para o cultivo do morangueiro no Brasil existe a dependência dos produtores em utilizar mudas importadas. Com intuito de reduzir essa dependência, uma das alternativas é o uso de mudas obtidas de plantas matrizes cultivadas em ambiente protegido. Porém, como essas mudas são produzidas no verão e plantadas no final desta estação ou no outono do ano seguinte, é necessário controlar o crescimento para diminuir os gastos energéticos das plantas. Objetivou-se com o trabalho avaliar o efeito de diferentes concentrações e épocas de aplicação de proexadione cálcio (ProCa) no controle de crescimento de mudas de morangueiro com torrão. O experimento foi realizado em casa de vegetação com mudas das cultivares Aromas e Camarosa, produzidas por enraizamento de estolões e mantidas em bandejas de poliestireno de 72 células em substrato casca de arroz carbonizada. O delineamento experimental utilizado foi inteiramente casualizado, com esquema fatorial 4x2 (4 concentrações de ProCa: 0, 100, 200 e 400 mg L-¹ x 2 épocas de aplicação: aos 20 e aos 30 dias após o período de plantio do estolão para enraizamento). Avaliaram-se nas mudas: sobrevivência de plantas, diâmetro de coroa, comprimento de pecíolo, área foliar total, área foliar específica, concentração de clorofila e massa seca da coroa e da parte aérea. A aplicação de ProCa aos 20 dias após o período de plantio do estolão para enraizamento nas concentrações de 200 e 400 mg L-¹ favorece a redução do comprimento do pecíolo em morangueiro "Aromas" e a aérea foliar total em morangueiro "Camarosa". A aplicação de ProCa a partir da concentração de 100 mg L-¹ reduz o crescimento vegetativo de mudas de morangueiro "Aromas" e"Camarosa" cultivadas em substrato.(AU)

Volatilização da amônia da ureia estabilizada com NBPT na adubação em cobertura da Urochloa ruziziensis / Ammonia volatilization from stabilized urea with NBPT of the topdressing fertilization in Urochloa ruziziensis

A ureia é o principal adubo nitrogenado utilizado em pastagens. Nesse fertilizante, uma das principais perdas de nitrogênio (N) está relacionada com a volatilização da amônia. O uso de produtos que proporcionam redução das perdas de N da ureia pode contribuir para aumentar a eficiência de uso desse nutriente em pastagens. Assim, objetivou-se determinar o efeito de fontes e doses de N aplicado parcelado em cobertura na Urochloa ruziziensis, nas perdas de N pela volatilização de amônia (N-NH3). O experimento foi conduzido em Santo Antônio de Goiás, GO, em Latossolo Vermelho Distrófico. Utilizou o delineamento experimental de blocos ao acaso, com quatro repetições em arranjo fatorial 2 x 5 x 5. Os tratamentos foram compostos pelas fontes de N: ureia comum e ureia com inibidor de urease, doses de N: 0, 50, 100, 200 e 300 kg ha-1 (parceladas em 5 aplicações) e cinco períodos de crescimento da forrageira: 14/11 a 13/12 (1º período), 14/12 a 12/01 (2º período), 13/01 a 11/02 (3º período), 24/03 a 22/04 (4º período), e 10/07 a 08/08 (5º período). O uso de ureia com inibidor de urease proporcionou reduções nas perdas de amônia por volatilização em relação à ureia comum. O aumento das doses de N proporcionou aumento da volatilização de N-NH3. Ocorreram maiores perdas de N pela volatilização da amônia nos períodos mais secos do ano.

Urea is the major nitrogen fertilizer used in pastures. In this fertilizer, one of the main nitrogen (N) losses is related to ammonia volatilization. The use of products that provide reduced losses of N from urea can help to increase the use efficiency of this nutrient in pastures. This study aimed to determine the effect of nitrogen sources and doses split applied at topdressing in Urochloa ruziziensis in the N losses by ammonia N-NH3 volatilization. The experiment was conducted in Santo Antônio de Goiás, GO, in a in a Brazilian Oxisol. It was used the randomized blocks experimental design, with four replications, in a factorial arrangement 2 x 5 x 5. Treatments were composed by N sources: common urea and urea with urease inhibitor, N rates: 0, 50, 100, 200 and 300 kg ha-1 (split in 5 applications) and five forage growth period: 11/14 to 12/13 (1º period), 12/14 to 01/12 (2º period), 01/13 to 02/11 (3º period), 03/24 to 04/22 (4º period) and 07/10 to 08/08 (5º period). The use of urea with urease inhibitor provided reductions in ammonia loss by volatilization in comparison to the common urea. Increasing N rates led to increased NNH3 volatilization. In drier periods of the year it was found greater losses of N by ammonia volatilization.