Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit

Abstract Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Resumo As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit / Compostos fenólicos e atividade antifúngica dos extratos acetato de etila e metanólico dos frutos maduros de Capsicum chinense Jacq

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit.

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Pregnancy in pigs: the journey of an early life.

Embryo mortality is responsible for greater losses in litter size in pigs. It is well known that pregnancy establishment is a complex process, and important changes occur continuously in both the corpora lutea and the endometrium, which varies depending on the pre-natal development phase: embryonic, pre-implantation or fetal stages. The placenta is a key organ responsible for the exchange of nutrients, metabolites and respiratory gases between mother and fetuses. The porcine placenta is diffuse, epitheliochorial, and placentation begins with implantation, which involves specialized cell adhesion and cell migration, leading to the attachment of the trophectoderm to the uterine endometrial lumen epithelium. The efficiency with which the placenta provides adequate amounts of nutrients and oxygen to the fetus is crucial for proper fetal growth and development. In the last decades, emphasis on selection for sow prolificacy has resulted in a substantial increase in the number of piglets born per litter, which had a direct effect on piglet quality, compromising birth weight and litter uniformity. Placental insufficiency will lead to fetal intrauterine growth restriction. This review addresses the main events of early embryo development, including preimplantation and implantation periods. In addition, placentation and its role on fetal development are covered, as well as intrauterine growth restriction, as it is a natural condition in the pig, with long lasting detrimental effects to the production chain.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum / Composição química e efeito inibitório in vitro dos óleos essenciais das cascas dos frutos de três espécies de Citrus e do limoneno sobre o crescimento micelial de Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 μL. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 μL) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 μL doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum.

Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 µL. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 µL) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 µL doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Blocking and remanence properties of weakly and highly interactive cobalt ferrite based nanoparticles.

We compare both magnetic blocking properties and remanence curves for dilute ferrofluid and powder samples of ferrite magnetic nanoparticles. Low field DC magnetization, AC susceptibility, isothermal remanent magnetization and DC demagnetization techniques are employed to investigate the role of interparticle magnetic interactions on the superparamagnetic relaxation, the magnetic anisotropy and on the super-spin-glass state in closely packed particles. The samples used herein are 3 nm sized spinel-type nanocrystals made of a cobalt ferrite core covered by a layer of maghemite on its outermost surface and can be obtained as aqueous colloidal dispersions thanks to this core-shell strategy. They show large anisotropy attributed to an enhanced surface contribution and the blocking temperature is shifted towards higher values as interparticle distance decreases. For all investigated diluted liquids and powder samples the frequency dependency of the peak temperature is well accounted by a Vogel-Fulcher law, with the insertion of a phenomenological temperature associated to the magnitude of interparticle dipolar interactions. The fractional change of the peak temperature per decade of frequency enlights the presence of interactions between particles in dilute liquids and of a spin-glass-like state in powder samples. The remanence curves always show global demagnetizing behavior, attributed to the combination of both spin surface disorder and interparticle dipolar interactions, the former being predominant in isolated nanoparticles and the latter in powder samples. However, in the most compacted powder, exchange interaction between surface ions of different particles becomes more pronounced and promotes an additive magnetizing effect.

Promising synergistic activity of fluconazole with bioactive Guttiferone-A and derivatives against non-albicans Candida species.

Natural products with diverse bioactivities and structures are an important source of novel chemicals with pharmaceutical potentials. Combinations of bioactive compounds with classical antimicrobial agents against drug-resistant or low-susceptible Candida spp. have been studied. Guttiferone-A and its derivatives were combined with fluconazole through the checkerboard method and tested against Candida spp. The results obtained, especially the Fractional Inhibitory Concentration Index (FICI) determined to the combinations, suggests promising results on the treatment of Candida infections, principally for species that present resistance or low antifungal susceptibility. The best result was seen for C. krusei, in which a synergic action of the association between fluconazole and Guttiferone-A resulted in a reduction of more than 100-fold in the alone inhibitory concentration (MIC) of fluconazole. Synergism was also noted in the association of fluconazole with the synthetic derivatives LFQM-79, LFQM-80 and LFQM-81 against C. glabrata, with reduction of up to four times in the alone IC of fluconazole. These results suggest the possibility of combined administration with reduction of doses and side effects of drugs conventionally used against Candida spp. and the promising therapeutic action of Guttiferone-A.

Effect of supplemental yeast culture and dietary starch content on rumen fermentation and digestion in dairy cows.

The objectives of this experiment were to evaluate the effect of feeding a culture of Saccharomyces cerevisiae on rumen metabolism and digestibility when cows are fed diets varying in starch content. Four lactating Holstein cows were assigned to a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Treatments were low starch (LS; 23% of diet DM) and no yeast culture (YC; LS-control), LS and 15 g of YC/d (LS-YC), high starch (HS; 29% of diet DM) and no YC (HS-control), and HS and 15 g of YC/d (HS-YC). Periods lasted 28 d, with the last 9 d for data collection. Days 20 to 24 were used to determine production, nutrient flow, and digestibility. On d 25, 3 kg of corn grain DM was placed in the rumen 1 h before the morning feeding, and yields of milk and milk components were measured after the challenge. Blood was sampled -1, 3, 7, and 11 h relative to the morning feeding on d 24 and 25. Rumen pH was measured continuously on d 24 and 25. Rumen papillae were collected on d 24 and 28 to quantify mRNA expression of select genes. Supplementing YC increased yields of milk (26.3 vs. 29.6 kg/d), energy-corrected milk (ECM; 26.5 vs. 30.3 kg/d), fat (0.94 vs. 1.08 kg/d), true protein (0.84 vs. 0.96 kg/d), and ECM/dry matter intake (1.15 vs. 1.30) compared with the control but did not affect dry matter intake (22.6 vs. 22.9 kg/d). Cows fed HS had increased milk true protein percentage (3.18 vs. 3.31%) and yield (0.87 vs. 0.94 kg/d) compared with cows fed LS. Feeding HS-YC increased the proportion of dietary N incorporated into milk true protein from 24.9% in the other 3 treatments to 29.6%. Feeding HS increased the concentration of propionate (21.7 vs. 32.3 mM) and reduced that of NH3-N (8.3 vs. 6.7 mg/dL) in rumen fluid compared with the control, and combining HS with YC in HS-YC tended to increase microbial N synthesis compared with LS-YC (275 vs. 322 g/d). Supplementing YC to cows fed HS reduced plasma haptoglobin and rumen lactate concentrations, increased mean rumen pH, reduced the time with pH <6.0, and prevented the decrease in rumen neutral detergent fiber digestion caused by HS. Cows fed HS had less total-tract digestion of organic matter (73.9 vs. 72.4%) because of reduced acid detergent fiber (57.6 vs. 51.7%) and neutral detergent fiber (60.9 vs. 56.7%) digestibility. Production performance after the challenge was similar to that before the challenge, and YC improved yield of ECM. After the challenge, supplementing YC tended to reduce rumen lactate concentration compared with the control and reduced haptoglobin in cows fed HS. Feeding HS but not YC increased expression in rumen papillae of genes for receptors (FFAR2 and FFAR3) and transporter (SLC16A3) of short-chain fatty acids but did not affect genes involved in transport of Na+/H+ or water or in inflammatory response. Supplementing YC to dairy cows improved lactation performance in diets containing low or high starch, and mechanisms might be partially attributed to improvements in rumen pH, digestion of fiber, microbial N synthesis, and reduction in acute phase response.

Effects of supplementing yeast culture to diets differing in starch content on performance and feeding behavior of dairy cows.

The objectives were to evaluate the effects of a culture of Saccharomyces cerevisiae (YC) on lactation performance of cows fed diets differing in starch content. Fifty-six Holstein cows at 42 d postpartum were blocked by parity and milk production and randomly assigned to 1 of 4 treatments, low starch (23% diet DM) and no YC (LS-control), low starch and 15 g/d of YC (LS-YC), high starch (29% diet DM) and no YC (HS-control), and high starch and 15 g/d of YC (HS-YC). The experiment lasted 14 wk. Blood was sampled twice weekly during the first 5 wk in the experiment. Feeding behavior was evaluated in 2 consecutive days when cows were 33 d in the experiment. On d 92 in the experiment, cows were challenged with 3 kg of corn grain DM immediately before the morning feeding. Blood was sampled in the first 12 h after the challenge. Rumen fluid was collected 5 h after the challenge, and pH, ammonia N, short-chain fatty acids, and lactate concentrations were quantified. Lactation performance was measured daily before and after the challenge. Supplementation with YC increased yields of 3.5% fat-corrected milk and energy-corrected milk by 2.2 and 2.0 kg/d, and the increments were observed in both low- and high-starch diets. Feeding HS tended to decrease milk fat content (LS = 3.88 vs. HS = 3.73%), but increased concentration (LS = 2.87 vs. HS = 3.00%) and yield (LS = 1.11 vs. HS = 1.20 kg/d) of milk true protein. Feeding YC increased yields of fat and true protein in milk by 100 and 60 g/d. Energy balance, body weight, and feed efficiency did not differ with treatments. Feeding HS reduced eating time (LS = 177 vs. HS = 159 min/12 h) and intermeal interval (LS = 103 vs. HS = 82 min), but tended to increase eating rate (LS = 139 vs. HS = 150 g/min). Interactions were detected between level of starch and YC for ruminating time, meal duration, and meal size because within LS, feeding YC increased ruminating time 23 min/12 h, but reduced meal duration 6 min/meal and meal size 0.7 kg/meal. Concentrations of glucose in plasma increased (LS = 62.1 vs. HS = 63.8 mg/dL), whereas those of urea N decreased (LS = 10.1 vs. HS = 9.4 mg/dL) with feeding HS compared with LS in the first 5 wk in the experiment, and the same responses were observed after the challenge with corn grain. After the challenge, rumen pH was less and short-chain fatty acid concentrations were greater in cows fed HS compared with those fed LS; however, supplementing YC to high-starch diets increased rumen pH (HS-control = 5.72 vs. HS-YC = 6.12) and reduced concentrations of lactate in rumen fluid (HS-control = 7.72 vs. HS-YC = 1.33 mM) and haptoglobin in plasma 28%. Feeding YC improved lactation performance irrespective of the level of dietary starch and reduced the risk of subacute rumen acidosis induced by a grain challenge when cows were fed a high-starch ration.

Effects of supplementation with ruminally protected choline on performance of multiparous Holstein cows did not depend upon prepartum caloric intake.

Objectives were to evaluate the effect of prepartum energy intake on performance of dairy cows supplemented with or without ruminally protected choline (RPC; 0 or 17.3 g/d of choline chloride; 0 or 60 g/d of ReaShure, Balchem Corp., New Hampton, NY). At 47 ± 6 d before the expected calving date, 93 multiparous Holstein cows were assigned randomly to 1 of 4 dietary treatments in a 2 × 2 factorial arrangement. Cows were fed energy to excess [EXE; 1.63 Mcal of net energy for lactation/kg of dry matter (DM)] or to maintenance (MNE; 1.40 Mcal of net energy for lactation/kg of DM) in ad libitum amounts throughout the nonlactating period. The RPC was top-dressed for 17 ± 4.6 d prepartum through 21 d postpartum (PP). After calving, cows were fed the same methionine-balanced diet, apart from RPC supplementation, through 15 wk PP. Liver was biopsied at -14, 7, 14, and 21 d relative to parturition. Cows fed EXE or MNE diets, respectively, consumed 40 or 10% more Mcal/d than required at 15 d before parturition. Cows fed the MNE compared with the EXE diet prepartum consumed 1.2 kg/d more DM postpartum but did not produce more milk (41.6 vs. 43.1 kg/d). Thus, PP cows fed the EXE diet prepartum were in greater mean negative energy balance, tended to have greater mean concentrations of circulating insulin, fatty acids, and ß-hydroxybutyrate, and had greater triacylglycerol in liver tissue (8.3 vs. 10.7% of DM) compared with cows fed the MNE diet prepartum. Cows fed RPC in transition tended to produce more milk (43.5 vs. 41.3 kg/d) and energy-corrected milk (44.2 vs. 42.0 kg/d) without increasing DM intake (23.8 vs. 23.2 kg/d) during the first 15 wk PP, and tended to produce more milk over the first 40 wk PP (37.1 vs. 35.0 kg/d). Energy balance of cows fed RPC was more negative at wk 2, 3, and 6 PP, but mean circulating concentrations of fatty acids and ß-hydroxybutyrate did not differ from those of cows not fed RPC. Despite differences in energy balance at 2 and 3 wk PP, mean concentration of hepatic triacylglycerol did not differ between RPC treatments. Feeding RPC reduced the daily prevalence of subclinical hypocalcemia from 25.5 to 10.5%, as defined by concentrations of total Ca of <8.0 mg/dL in serum in the first 7 d PP. Pregnancy at first artificial insemination tended to be greater for cows fed RPC (41.3 vs. 23.6%), but the proportion of pregnant cows did not differ by 40 wk PP. Heifers born from singleton calvings from cows fed RPC tended to experience greater daily gain between birth and 50 wk of age than heifers from cows not supplemented with RPC. Feeding RPC for approximately 38 d during the transition period tended to increase yield of milk for 40 wk regardless of amount of energy consumed during the pregnant, nonlactating period.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Role of MTHFR C677T and MTR A2756G polymorphisms in thyroid and breast cancer development.

Folate metabolism is essential for DNA synthesis and repair. Alterations in genes that participate in folate metabolism can be associated with several types of malignant neoplasms, including thyroid and breast cancer. In the present case-control study, we examined the association between methylenetetrahydrofolate reductase (MTHFR C677T, rs1801133) and methionine synthase (MTR A2756G, rs1805087) polymorphisms and risk for thyroid and breast cancer. Polymerase chain reaction-restriction fragment length technique was used to determine the specific genotypes in the genes of interest. Statistical analysis was performed by multiple logistic regression test. We found an association between MTHFR C677T polymorphism and risks to both thyroid (OR = 2.50; 95%CI = 1.15-5.46; P = 0.02) and breast cancer (OR = 2.53; 95%CI = 1.08-5.93; P = 0.03). Tobacco consumption and high body mass index were also associated with thyroid cancer. In addition, increased age (≥50 years) and alcohol consumption were found to be associated with breast cancer. Our results indicated that MTHFR C677T is significantly associated with thyroid and breast cancer risks. Thus, these factors may be used as potential prognostic markers for thyroid and breast cancers.

Patient perceptions concerning clinical trials in oncology patients.

BACKGROUND: Clinical trials are critical to scientifically evaluate promising new therapies in oncology, but patient accrual to these studies is persistently low. Patient preference plays an important role in enrollment in these trials. We performed this survey to evaluate the perceptions of newly diagnosed oncology patients about clinical trials and the reasons why they wish to or not to participate in these trials. METHODS: Patients were given a ten question survey reflective of their attitudes regarding clinical trials as a treatment option at their initial visit. The self-directed questionnaire was scored on an ordinate scale from strongly agree [1] to strongly disagree [5]. RESULTS: Ninety three patients were surveyed in the cancer specific multispecialty clinics in an academic center. Our patients expected their providers to discuss all information relating to clinical trials and eligibility at the first visit (65.4% agree and 15.4% neutral, p < 0.0001). Patients felt their privacy and safety would be safeguarded in the University sponsored trials (56.8% agree, and 25.7% neutral, p < 0.0001). Over 80% patients showed their unwillingness to participate in randomized clinical trials (disagree 61%, neutral 19.5%, p < 0.001). Patients also showed less likelihood to participate in clinical trials as a first treatment option (48.7% disagree and 28.9% neutral, p0.0161), but were willing to consider participating in a clinical trial if the conventional treatment failed. Industry sponsored trials, phase 1 trials, investigator initiated trials with the involved tests and time commitment and altruistic reasons did not significantly deviate from the mean preference analyzed using Fisher's exact test analysis. CONCLUSIONS: Patients consider the option of clinical trials as important in their treatment, and expect to be informed by their oncologist about such trials. Newly diagnosed cancer patients perceive randomization and first line trials negatively. Since randomization data provides new standards of care and hope for improved treatment, patients and their families must be educated of their importance.

Cancer stem cells in head and neck cancer: A Mini Review.

Head and neck cancer (HNC) is a multifaceted and genomically complex disease and rapidly emerging preclinical and clinical studies have provided a broader landscape of signaling. It is being realized that intra-tumor heterogeneity, genetic and epigenetic mutations considerably challenge wide ranging therapeutics and patients frequently develop locoregional recurrences, second primary tumours and distant metastases. Using high-throughput technologies, it has been revealed that existence of different subpopulations of cells within tumor mass with different phenotypic and functional properties with distinct tumour-initiating potential is responsible to HNC resistance. In light of accumulating evidence reported in recent years, it is now known that different intracellular proteins and cell surface markers have been used to study CSCs. This review provides an overview of CSC biomarkers in HNC treatment and their potential as therapeutic targets in improving the diagnosis, prognosis and treatment of HNC patients for new therapeutic strategies with information about estimation of prognosis and treatment decision. Further studies regarding biomarkers are necessary to determine the specific role of CSCs in HNC which could be useful in development of new therapeutic strategies to eliminate CSCs and maximize clinical outcome. Furthermore, CD44 still need more research in HNC once the studies show contradictions. Studies using lineage tracing and deep sequencing will provide a comprehensive understanding of CSC model and extent to which it is accountable for resistance against therapeutics and carcinogenesis.

Fenóis totais, atividade antioxidante e inibição da enzima tirosinase de extratos de Myracrodruon urundeuva Fr. All. (Anacardiaceae) / Total phenolics, antioxidant activity and tyrosinase enzyme inhibition by Myracrodruon urundeuva Fr. All. (Anacardiaceae) extracts

RESUMOOs radicais livres e outros oxidantes demonstram ser parcialmente responsáveis pelo envelhecimento e pelas doenças degenerativas associadas à produção de espécies reativas de oxigênio. Além disto, os compostos fenólicos são reconhecidamente detentores de elevada atividade antioxidante, que geralmente estão envolvidos em tratamentos de problemas de pigmentação da pele, que resultam em hiperpigmentações. Sendo assim este trabalho foi realizado para avaliar o conteúdo de fenóis totais, atividade antioxidante e a capacidade de inibição da tirosinase dos extratos das folhas e cascas do caule da espécie Myracrodruonurundeuva Fr. All. Para determinação do conteúdo de fenóis totais dos extratos orgânicos de M. urundeuva utilizou-se o reativo Folin-Ciocalteau, na avaliação da atividade antioxidante empregando o radical livre DPPH, enquanto que os testes de inibição da enzima tirosinase foram realizados utilizando L-tirosina. A concentração de fenóis totais foi de 77 mg EAG g-1 e 194 mg EAG g-1 nos extratos hexânico e metanólico das folhas e de 45 mg EAG g-1 e 193 mg EAG g-1 nos extratos hexânico e metanólico das cascas do caule. O potencial antioxidante dos extratos indicaram que o extrato metanólico das cascas do caule (10,9 ± 0,5 µg mL-1), em comparação ao hexânico (12,9 ± 0,2 µg mL-1) e ao BHT (220 ± 7,0 µg mL-1), possui atividade antioxidante levemente mais acentuada. No ensaio de inibição da tirosinase, o extrato metanólico das cascas do caule demonstrou a inibição da enzima em 42% após uma hora.

ABSTRACTFree radicals and other oxidants had demonstrated to be partially responsible for aging and for degenerative diseases associated with the production of reactive oxygen species. In addition, the phenolic compounds are recognized as holders of high antioxidant activity, which usually are involved in the treatment of pigmentation problems, resulting in hyper pigmentation. So this study was carried out in order to evaluate the content of total phenols, antioxidant activity and capacity inhibition of tyrosinase in extracts from leaves and stem barks of the species Myracrodruon urundeuva Fr. All. In order to determine the content of total phenols from organic extracts of M. urundeuva, a Folin-Ciocalteau reagent was employed in the evaluation of antioxidant activity,using the DPPH free radical, while the enzyme tyrosinase inhibition tests were performed using L-tyrosine. The amount of total phenols was 77 mg EAG g-1 and 194 in the hexane and methanolic extracts from the leaves and 45 mg EAG g-1 and 193 mg EAG g-1in the hexane and methanolic extracts of the stem barks. The antioxidant potential of extracts indicated that the methanolic extract of stem bark (10.9 ± 0.5 µg mL-1), when compared to the hexane (12.9 ± 0.2 µg mL-1) and BHT (220 ± 7.0 µg mL-1), presented a slightly higher antioxidant activity. For the tyrosinase inhibitiontrial , the methanolic extract ofstem barks indicated anenzyme inhibition of 42% after an hour.

Effects of supplemental progesterone after artificial insemination on expression of interferon-stimulated genes and fertility in dairy cows.

The objectives of the current study were to evaluate the effects of supplemental progesterone after artificial insemination (AI) on expression of IFN-stimulated genes (ISG) in blood leukocytes and fertility in lactating dairy cows. Weekly cohorts of Holstein cows were blocked by parity (575 primiparous and 923 multiparous) and method of insemination (timed AI or AI on estrus) and allocated randomly within each block to untreated controls, a controlled internal drug release (CIDR) containing 1.38g of progesterone from d 4 to 18 after AI (CIDR4), or a CIDR on d 4 and another on d 7 after AI and both removed on d 18 (CIDR4+7). Blood was sampled to quantify progesterone concentrations in plasma and mRNA expression in leukocytes for the ubiquitin-like IFN-stimulated gene 15-kDa protein (ISG15) and receptor transporter protein-4 (RTP4) genes. Pregnancy was diagnosed on d 34±3 and 62±3 after AI. Treatment increased progesterone concentrations between d 5 and 18 after AI in a dose-dependent manner (control=3.42, CIDR4=4.97, and CIDR4+7=5.46ng/mL). Cows supplemented with progesterone tended to have increased luteolysis by d 19 after AI (control=17.2; CIDR4=29.1; CIDR4+7=30.2%), which resulted in a shorter AI interval for those reinseminated after study d 18. Pregnancy upregulated expression of ISG in leukocytes on d 19 of gestation, but supplementing progesterone did not increase mRNA abundance for ISG15 and RTP4 on d 16 after insemination and tended to reduce mRNA expression on d 19 after AI. For RTP4 on d 19, the negative effect of supplemental progesterone was observed only in the nonpregnant cows. No overall effect of treatment was observed on pregnancy per AI on d 62 after insemination and averaged 28.6, 32.7, and 29.5% for control, CIDR4, and CIDR4+7, respectively. Interestingly, an interaction between level of supplemental progesterone and method of AI was observed for pregnancy per AI. For cows receiving exogenous progesterone, the lower supplementation with CIDR4 increased pregnancy per AI on d 62 in cows inseminated following timed AI (CIDR4=39.2; CIDR4+7=27.5%); in those inseminated following detection of estrus, however, the use of a second insert on d 7 resulted in greater pregnancy per AI (CIDR4=26.9; CIDR4+7=31.5%). Pregnancy loss did not differ among treatments. Supplemental progesterone post-AI using a single intravaginal insert on d 4 was beneficial to pregnancy in cows inseminated following timed AI, but incremental progesterone with a second insert on d 7 did not improve fertility of dairy cows.

Karyotypic diversity in a population of Bryconamericus aff. iheringii (Characidae).

Bryconamericus comprises 56 species distributed into three groups, on the basis of the position and shape of the maxillary teeth: B. exodon, B. microcephalus and B. iheringii groups. Few cytogenetic data are available for this genus, but the diploid number of 52 chromosomes is quite common, although the karyotypic variability is extensive. This study aimed to characterize a population of B. aff. iheringii and thus contribute more cytogenetic information and better understanding of the structure and karyotypic evolution of this genus. We found 6 cytotypes with different NOR patterns: cytotype I showed a karyotype formula of 12m+10sm+16st+14a (FN = 90) and single NORs; cytotype II with 18m+14sm+10st+10a (FN = 94) and cytotype III with 20m+18sm+4st+10a (FN = 94), showing both single and multiple NORs; cytotype IV with 20m+14sm+12st+6a (NF = 98), cytotype V with 22m+18sm+8st+4a (FN = 100) and cytotype VI with 18m+24sm+6st+4a (FN = 100), all with multiple NORs. Cytotype I is the most different in relation to FN and NOR pattern, and can be regarded as belonging to another species of the genus Bryconamericus, living in sympatry in Três Bocas Stream. The remaining cytotypes may have been generated by crosses between them and by pericentric inversions. Meiotic cells were also analyzed and showed that despite the high karyotypic variability, chromosome pairing occurred normally. The great variability found in B. aff. iheringii may be related to a high degree of polymorphism; nevertheless, the possibility of occurrence of more than one species in this location is not ruled out, demonstrating the need for conservation of the Três Bocas Stream.

Physical mapping of 18S rDNA and heterochromatin in species of family Lygaeidae (Hemiptera: Heteroptera).

Analyses conducted using repetitive DNAs have contributed to better understanding the chromosome structure and evolution of several species of insects. There are few data on the organization, localization, and evolutionary behavior of repetitive DNA in the family Lygaeidae, especially in Brazilian species. To elucidate the physical mapping and evolutionary events that involve these sequences, we cytogenetically analyzed three species of Lygaeidae and found 2n (♂) = 18 (16 + XY) for Oncopeltus femoralis; 2n (♂) = 14 (12 + XY) for Ochrimnus sagax; and 2n (♂) = 12 (10 + XY) for Lygaeus peruvianus. Each species showed different quantities of heterochromatin, which also showed variation in their molecular composition by fluorochrome staining. Amplification of the 18S rDNA generated a fragment of approximately 787 bp. The alignment of the consensus sequence with sequences from other species of Heteroptera deposited in the GenBank revealed a similarity of 98% with small differences. Fluorescent in situ hybridization with the 18S rDNA fragment revealed that this ribosomal gene was located in 1 autosomal pair at different positions in the three species. No cytogenetic data are available for these Brazilian species. The basal number and the possible chromosomal changes that occurred among the different species, as well as the evolution of these DNA sequences, are discussed.