Guarana (Paullinia cupana) as a potential tool for mesenchymal stromal cells priming in regenerative medicine

Mesenchymal stromal cells (MSCs) have therapeutic potential due to their abilities of differentiation, immunomodulation, and migration to injured tissues, potentiating such effects when cells are activated. Guarana (Paullinia cupana) is a tropical plant species found in South America that is known for its antioxidant, stimulant, and cicatricial effects. The guarana extract is composed of many substances and caffeine is the main component. The objective was to evaluate the effects of guarana and caffeine on MSCs. After the initial characterization, MSCs were treated with Paullinia cupana (10, 100, and 1000 μg/mL) or caffeine (0.4, 4, and 40 μg/mL) for 24 h. MSCs treatment with 1000 μg/mL guarana increased cell polarity, viability, cell migration to chemoattractant, antioxidant potential, and liberation of extracellular vesicles (EVs), while it reduced the levels of autophagy. MSCs treated with 100 and 1000 μg/mL guarana or 40 μg/mL caffeine showed a decrease of cell proliferation. No treatment affected the cellular area and cell cycle of MSCs. The study shows in vitro evidence that guarana could be a promising alternative for activating MSCs to promote better cellular products for future clinical therapies.

Quantificação do colágeno dérmico equino por duas técnicas morfométricas: contagem de pontos e segmentação de cor / Quantification of equine dermal collagen by two morphometric techniques: point counting and color segmentation

Os colágenos tipos I e III apresentam diferentes tonalidades de birrefringência em cortes histológicos corados com Picrosirius red e analisados em microscópio sob luz polarizada. Com base nessa propriedade, os colágenos podem ser quantificados por histomorfometria. Entretanto, são muitas as variáveis que podem afetar a distribuição das cores na imagem histológica, e a escolha adequada dos parâmetros de análise têm grande influência no resultado final. O objetivo deste trabalho foi comparar a quantificação histomorfométrica de colágeno em pele equina pela morfometria por contagem de pontos e pela segmentação de cor com diversas configurações, a fim de se determinar o melhor método de avaliação. Para a morfometria por contagem de pontos, foram utilizadas três gratículas diferentes (391, 588 e 792 pontos de interseções) e, para a segmentação de cor, seis combinações de hue e brightness no software ImageJ. Os valores foram submetidos ao teste de Friedman, seguido pelo teste de Tukey com 5% de significância. Os resultados demonstraram que a quantificação dos colágenos na gratícula de 792 pontos foi equivalente aos resultados da segmentação de cor com brightness de 1-255 e hue de 0-42 e 43-120 para os colágenos tipos I e III, respectivamente. Dessa forma, conclui-se que a análise automática da segmentação de cor, utilizando configuração adequada para brightness e hue, pode substituir a morfometria por contagem de pontos de forma confiável e segura.(AU)

The types I and III collagens present different tonalities of birefringence in histological sections stained with Picrosirius red, that can be analyzed under a polarized light microscope. Based on this property, collagens can be quantified by histomorphometry. However, many variables can affect the color distribution in the histological image, and the appropriate choice of the analysis parameters have a significant influence on the final result. The objective of this study was to compare the histomorphometric quantification of collagen in the equine skin by counting points planimetry and color segmentation with different configurations to determine the best method of evaluation. For planimetry, three different graticules (391, 588 and 792 intersections) were used and, for color segmentation, six combinations of hue and brightness in ImageJ software. The values were submitted to the Friedman test followed by Tukey with 5% significance. The results showed that the quantification of collagens in the graticule of 792 intersections was equivalent to the results of color segmentation with a brightness of 1-255 and hue of 0-42 and 43-120 for collagens type I and III, respectively. Automatic analysis of the color segmentation, using suitable configuration for brightness and hue can replace the counting points planimetry reliably and safely.(AU)

Rendimento do coentro (Coriandrum sativum L) adubado com esterco bovino em diferentes doses e tempos de incorporação no solo / Yield of the coriander [Coriandrum sativum (L)] fertilized with manure at different doses and incorporation times in the soil

RESUMO O uso de esterco bovino é de suma importância para os agricultores familiares que produzem coentro na região de Mossoró-RN, pois esse insumo é amplamente disponível e utilizado pelos agricultores. Objetivando-se avaliar o Rendimento do coentro fertilizado com esterco bovino em diferentes doses e tempos de incorporação ao solo foi conduzido um experimento no período de setembro a novembro de 2011, na Fazenda Experimental Rafael Fernandes, da Universidade Federal Rural do Semi-Árido - UFERSA, Brasil. Os tratamentos consistiram da combinação de quatro doses de esterco bovino incorporadas ao solo: 15,0; 30,0; 45,0 e 60,0 t ha-1 em base seca, com quatro tempos de incorporação: 28; 49; 64 e 80 dias antes da semeadura do coentro - DAS, mais um tratamento controle (ausência de adubação). O delineamento experimental utilizado foi em blocos completos casualizados com os tratamentos arranjados em esquema fatorial 4 x 4 + 1 com 3 repetições. A cultivar de coentro utilizado foi a Verdão e as variáveis determinadas foram altura e número de hastes por planta e rendimento de coentro. O coentro respondeu à aplicação de esterco bovino, produzindo rendimentos máximos de 6453 e 6349 kg ha-1 de massa verde, com a dose de 60 t ha-1 e aos quarenta e seis dias de incorporação antes da semeadura, respectivamente.

ABSTRACT The use of manure is very important to family farmers who produce coriander in the region of Mossoró-RN, because this input is widely available and used by farmers. Aiming to evaluate the yield of the coriander fertilized with manure at different doses and incorporation times in the soil, a trial was conducted from September to November 2011, at the Experimental Farm Rafael Fernandes in the Universidade Federal Rural do Semi-Árido (UFERSA), Mossoró-RN, Brazil. The treatments consisted on the combination of four levels of manure incorporated into the soil: 15.0; 30.0; 45.0 and 60.0 t ha-1 at dry basis, with four incorporation times: 28; 49; 64 and 80 days before the coriander"s sowing - DAS, plus a control treatment (without fertilization). The experimental design was a randomized complete block with treatments arranged in a 4 x 4 + 1, with three replications. The coriander planted was the "Verdão" and the specific variables were height and number of stalks per plant and yield of the coriander. The coriander responded to the application of the manure, producing maximum yields of 6453 and 6349 kg ha-1 of fresh mass, with the dose of 60 t ha-1 and at forty-six days of incorporation before the sowing, respectively.

Avaliação biométrica de caracteres do melão de São Caetano (Momordica charantia L) / Biometric evaluation of the characters of the melon from Sao Caetano (Momordica charantia L)

RESUMOO melão de São Caetano, planta trepadeira originaria do leste indiano e sul da Índia, é encontrada no bioma caatinga. Neste trabalho objetivou-se avaliar a biometria dos frutos e o número de sementes por fruto do melão de São Caetano. Os frutos foram coletados em área de vegetação nativa do campus da Universidade Federal Rural do Semi-Árido (UFERSA), situada no município de Mossoró, RN, seguindo para o laboratório de fitotecnia para a determinação das seguintes características: comprimento, diâmetro e peso dos frutos, bem como o número de sementes por fruto. As análises descritivas e gráficas foram realizadas utilizando o pacote estatístico software R-project. As variáveis, comprimento e diâmetro apresentaram baixa amplitude de variação e bons valores de coeficiente de variação. Já o peso do fruto e o número de sementes por fruto mostraram alta amplitude de variação. Verificou-se alto grau de simetria e distribuição mesocúrtica para todas as características avaliadas nesse estudo. Constatou-se também a correlação linear simples positiva significativa entre o comprimento e o diâmetro, e não significativa entre o peso e o número de sementes por fruto, e que os dados referentes a todas as características do fruto do melão de São Caetano avaliadas neste trabalho se ajustaram à distribuição normal de probabilidade, o que permite a realização de inferências com base estatística.

ABSTRACTThe melon from Sao Caetano is a climbing plant, originally from India and South East India and is found in the savanna biome. This study aimed to evaluate the biometric fruit and the number of seeds per fruit of the melon Sao Caetano. The fruits were collected inside a native vegetation area of the campus at the Universidade Federal Rural do Semi-Árido (UFERSA), located in the city of Mossoró, RN. They were then taken to the laboratory of crop science, for the determination of the following characteristics: length, diameter and fruit weight as well as number of seeds per fruit. The descriptive and graphical analyzes were performed by using the statistical package R -project software. The length and diameter variables exhibited a low amplitude variation and a good value for the coefficient of variation. However, the fruit weight and number of seeds per fruit showed a high amplitude variation. There was a high degree of symmetry and a flattened distribution for all traits evaluated in this study. It was also found significant positive simple linear correlation between the length and diameter, and no significant difference between the weight and number of seeds per fruit. The data related to all features of the São Caetano melon fruit evaluated in this study were adjusted to normal distribution of probability which allows the performance of interferences with statistical basis.

Rendimento de coentro (Coriandrum sativum L. ) em sistema de adubação verde com a planta jitirana (Merremia aegyptia L. ) / Yield of coriander (Coriandrum sativum L. ) in a system of green manure with the plant scarlet starglory (Merremia aegyptia L.)

O coentro é a hortaliça mais utilizada como condimento na região de Mossoró-RN. Um experimento foi conduzido na Fazenda Experimental Rafael Fernandes da Universidade Federal Rural do Semi-Árido - UFERSA, no período de maio a junho de 2010, com o objetivo de avaliar o rendimento de coentro sob diferentes quantidades da planta trepadeira jitirana, incorporada ao solo como adubo verde. O delineamento experimental usado foi o de blocos completos casualizados com sete tratamentos e três repetições. Os tratamentos consistiram da incorporação de sete quantidades de jitirana (3,0; 6,0; 9,0; 12,0; 15,0; 18,0 e 21,0 t ha-1 de matéria seca de jitirana). A cultivar de coentro plantado foi a Verdão. As características avaliadas foram: altura e número de hastes por planta, rendimento e massa seca da parte aérea. O melhor desempenho agronômico do coentro foi observado na quantidade de 21,0 t ha-1 de jitirana. Para cada tonelada de jitirana incorporada ao solo observa-se um rendimento de coentro de 395 kg ha-1.

The ciliandro is the vegetable more used as seasoning in the area at Mossoró-RN. An experiment was conducted at the Experimental Farm Rafael Fernandes of the Universidade Federal Rural do Semi-Árido - UFERSA in the period from may to june 2010, with the objective of evaluating the coriander yield in under different amounts of plant clambering scarlet starglory, incorporated into the soil. The experimental design was randomized complete blocks with eight treatments and three replicates. The treatments were combinations of eight amounts of scarlet starglory (3.0; 6.0; 9.0; 12.0; 15.0; 18.0 and 21.0 t ha-1 dry matter). The coriander cultivar planted was Verdão. The characteristics evaluated in the coriander were: plant height and number of stalks per plant, yield and dry matter mass of shoots. The best agroconomic performance lettuce was observed in the amount of 21,0 t ha-1 scarlet starglory. For each fresh or dry scarlet starglory ton incorporated into the soil, it was observed a mean yield of coriander mass of 395 kg ha-1.

Utilização do óleo de alho e da amoxilina, metronidazol e omeprazol no controle de Helicobacter spp. em cães / Use of garlic oil and amoxicillin, metronidazole, and omeprazol in the control of Helicobacter spp. in dogs

Avaliaram-se a eficácia do óleo de alho e da terapia tripla (amoxicilina, metronidazol e omeprazol) no tratamento de 21 cães infectados por Helicobacter spp., que apresentavam alterações histológicas nas biopsias endoscópicas da mucosa gástrica e reação positiva ao teste de urease. Os animais foram distribuídos, aleatoriamente, em três grupos de sete cães, os quais receberam os seguintes tratamentos: grupo 1 - cápsulas vazias; grupo 2 - 500mg de óleo de alho em cápsulas, diariamente, por um período de 30 dias; grupo 3 - amoxicilina, metronidazol e omeprazol, respectivamente, nas doses de 20mg/kg a cada 12 horas, 25mg/kg e 20mg/kg a cada 24 horas, durante 15 dias. Ao final dos tratamentos, os cães foram submetidos à endoscopia com realização de biopsias da mucosa gástrica. O tratamento com amoxicilina, metronidazol e omeprazol resultou em erradicação de Helicobacter spp. tanto na região fúndica quanto na pilórica. No grupo 2, houve redução da degeneração glandular na região fúndica em dois animais e em outros dois na pilórica. O tratamento com óleo de alho não foi eficaz em erradicar Helicobacter spp., apenas reduziu a sua colonização em quatro dos animais tratados.

The efficacy of garlic oil and triple therapy (amoxicillin, metronidazole, and omeprazol) were evaluated in the treatment of 21 dogs infected by Helicobacter spp., which presented histological alterations of the gastric mucosa according to endoscopic biopsies and positive reaction to urease test. The animals were randomly distributed into three groups of seven dogs each, and received the following treatment, group 1 - empty capsules; group 2 - 500mg of garlic oil capsules daily for a period of 30 days; and group 3 - amoxicillin, metronidazole, and omeprazol, in doses of 20mg/kg every 12 hours, 25mg/kg and 20mg/kg every 24 hours, respectively, for 15 days. By the end of the treatment, the dogs were subjected to new endoscopic procedure with gastric mucosa biopsies. Treatment with amoxicillin, metronidazole, and omeprazol resulted in eradication of Helicobacter spp. both in fundic and pyloric regions. The garlic oil treatment reduced glandular degeneration in the fundic region in two animals and in the pylorus region in two other animals, and it was inefficient in eradicating Helicobacter spp. in dogs as it only reduced the colonization in some of the treated animals.

Effect of alfa-difluormethylornithine on Anastrepha fraterculus (Diptera, Tephritidae) ovary size / Efeito da alfa-difluorometilornitina nas dimensões dos ovários em Anastrepha fraterculus (Diptera, Tephritidae)

Ovarian sizes (length and width) were measured in young females of Anastrepha fraterculus (Wiedemann) (Diptera, Tephritidae) subjected or not to the inhibitor α -difluormethylornithine (α -DFMO). The most effective concentration of α -DMFO used was 50 mM and the ovarian measurements (length and width) of the treated females were smaller than those of females not treated with α -DMFO. These data may suggest some relationship between ornithine decarboxylase (ODC) and sexual maturation in A. fraterculus.

As dimensões dos ovários (comprimento e largura) foram mensuradas em fêmeas jovens da Anastrepha fraterculus (Wiedemann) (Diptera, Tephritidae) submetidas ou não ao inibidor α -difluormetilornitina (α -DFMO). A concentração mais efetiva de α -DMFO utilizada foi 50 mM e as medidas (comprimento e largura) das fêmeas tratadas com o inibidor foram menores que as fêmeas não tratadas com inibidor α -DMFO. Estes dados podem sugerir uma relação entre ornitina descarboxilase (ODC) e maturação sexual em A. fraterculus.

On the absence of mutations in nucleotide excision repair genes in sporadic solid tumors

In general, stochastic tumors show genomic instability associated with the proliferation of DNA point mutations, that is, a mutator phenotype. This feature cannot be explained by a dysfunctional mismatch repair alone, and indicates that nucleotide excision repair (NER) and/or base excision repair should be suppressed. However, mutations in NER genes are not causally implicated in the oncogenesis of sporadic solid tumors, according to the Cancer Gene Census at http://www.sanger.ac.uk/genetics/CGP/Census/. This brings up an apparent paradox: how to explain the recurrent non-existence in NER genes of somatic mutations causally related to cancer? In a recent study, we have shown that the origin of point mutations in cancer cell genomes can be explained by a structurally conserved NER with a functional disorder generated from its entanglement with a disabled apoptosis gene network. In the present study, we further characterize NER gene network properties and show that it has a highly connected architecture. This feature suggests that the absence of mutations in NER genes in sporadic solid tumors is a result of their participation in many essential cellular functions.

Behavioral deficits in sepsis-surviving rats induced by cecal ligation and perforation

Sepsis and its complications are the leading causes of mortality in intensive care units, accounting for 10-50 percent of deaths. Intensive care unit survivors present long-term cognitive impairment, including alterations in memory, attention, concentration, and/or global loss of cognitive function. In the present study, we investigated behavioral alterations in sepsis-surviving rats. One hundred and ten male Wistar rats (3-4 months, 250-300 g) were submitted to cecal ligation and puncture (CLP), and 44 were submitted to sham operation. Forty-four rats (40 percent) survived after CLP, and all sham-operated animals survived and were used as control. Twenty animals of each group were used in the object recognition task (10 in short-term memory and 10 in long-term memory), 12 in the plus-maze test and 12 in the forced swimming test. Ten days after surgery, the animals were submitted individually to an object recognition task, plus-maze and forced swimming tests. A significant impairment of short- and long-term recognition memory was observed in the sepsis group (recognition index 0.75 vs 0.55 and 0.74 vs 0.51 for short- and long-term memory, respectively (P < 0.05). In the elevated plus-maze test no difference was observed between groups in any of the parameters assessed. In addition, sepsis survivors presented an increase in immobility time in the forced swimming test (180 vs 233 s, P < 0.05), suggesting the presence of depressive-like symptoms in these animals after recovery from sepsis. The present results demonstrated that rats surviving exposure to CLP, a classical sepsis model, presented recognition memory impairment and depressive-like symptoms but not anxiety-like behavior.

The effects of constant and alternating temperatures on the reproductive potential, life span, and life expectancy of Anastrepha fraterculus (Wiedemann) (Dipteria: Tephritidae)

Ovarian development, oviposition, larval eclosion, ornithine decarboxylase (ODC) activity, ovarian, testis and ejaculatory apodeme measurements (length, width, and area), and the number of spermatozoa of Anastrepha fraterculus (Wiedemann) were analyzed at alternating (20º/6ºC and 20º/13°C) and constant (6°C; 25°C) temperatures. Life span and life expectancy were also analyzed for both genders. All the results suggest that temperature, especially alternating temperatures, increase not only male and female reproductive potential but also their life span and life expectancy. These changes can be a powerful strategy triggered by A. fraterculus as a means to survive the stressful temperature conditions found in winter in the apple production region in Brazil, enabling this species to increase its population density and cause apple damage when spring begins

Intercellular contact-dependent survival of human A549, NCI-H596 and NCI-H520 non-small cell lung carcinoma cell lines

In the present study, we examined the relationship between cell phenotype and cell survival of three human non-small cell lung carcinoma cell lines (A549, NCI-H596 and NCI-H520). Cells in exponential growth at various densities were incubated for 24 h at 37ºC in a 5 percent CO2 humidified atmosphere and then exposed to UV radiation for 1 min (256 nm, 40 W, source-to-target distance 100 cm). After two days the surviving cells were quantified by sulforhodamine á staining and DNA fragmentation assay. The differences in UV sensitivity at 60 x 10(3) cells/cm(2) among the cell lines were not related to the proliferative state of the cells but to the extent of intercellular contact. In contrast to A549 and NCI-H596, irradiated NCI-H520 cells presented lower DNA fragmentation and an aggregated cell culture phenotype even prior to confluence, suggesting that a contact-effect mechanism provides further protection against UV radiation

Retinol-induced changes in the phosphorylation levels of histones and high mobility group proteins from Sertoli cells

Chromatin proteins play a role in the organization and functions of DNA. Covalent modifications of nuclear proteins modulate their interactions with DNA sequences and are probably one of the multiple factors involved in the process of switch on/off transcriptionally active regions of DNA. Histones and high mobility group proteins (HMG) are subject to many covalent modifications that may modulate their capacity to bind to DNA. We investigated the changes induced in the phosphorylation pattern of cultured Wistar rat Sertoli cell histones and high mobility group protein subfamilies exposed to 7 µM retinol for up to 48 h. In each experiment, 6 h before the end of the retinol treatment each culture flask received 370 KBq/ml [32P]-phosphate. The histone and HMGs were isolated as previously described [Moreira et al. Medical Science Research (1994) 22: 783-784]. The total protein obtained by either method was quantified and electrophoresed as described by Spiker [Analytical Biochemistry (1980) 108: 263-265]. The gels were stained with Coomassie brilliant blue R-250 and the stained bands were cut and dissolved in 0.5 ml 30 per cent H2O2 at 60oC for 12 h. The vials were chilled and 5.0 ml scintillation liquid was added. The radioactivity in each vial was determined with a liquid scintillation counter. Retinol treatment significantly changed the pattern of each subfamily of histone and high mobility group proteins.