Chemical characterization and effects of volatile oil of Alpinia zerumbet on the quality of collagen deposition and caveolin-1 expression in a muscular fibrosis murine model

Abstract This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Resumo Este estudo avaliou o efeito do óleo volátil de Alpinia zerumbet (OVAz) na expressão do gene da caveolina-1 e na fibrose muscular. Os ratos foram imobilizados para induzir a fibrose do músculo gastrocnêmio, e foram tratados com OVAz. A qualidade do colágeno foi avaliada com histologia e à expressão do gene caveolina-1 (CAV-1) foi avaliada usando qPCR. A análise histomorfológica indicou uma redução significativa no perímetro, largura e intensidade do colágeno nos grupos tratados. Os resultados dos níveis de expressão sugeriram diminuição nos grupos de lesão e em dois grupos de tratamento (0,0115 µg/g e 0,009 µg/g). No entanto, com a menor concentração (0,0065 µg/g), não foi observada diferença significativa, apresentando níveis semelhantes aos encontrados em tecido saudável. O uso do OVAz foi eficaz para reverter as alterações do colágeno causadas pela fibrose, e sua menor concentração apresentou uma possível tendência de aumento na expressão do CAV-1. Portanto, os resultados mostraram que o OVAz tem potencial para ser uma alternativa não invasiva e de baixo custo para auxiliar no tratamento da fibrose muscular.

Chemical characterization and effects of volatile oil of Alpinia zerumbet on the quality of collagen deposition and caveolin-1 expression in a muscular fibrosis murine model / Caracterização química e efeitos do óleo volátil de Alpinia zerumbet na qualidade da deposição do colágeno e expressão da caveolina-1 em modelo murino de fibrose muscular

Abstract This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Resumo Este estudo avaliou o efeito do óleo volátil de Alpinia zerumbet (OVAz) na expressão do gene da caveolina-1 e na fibrose muscular. Os ratos foram imobilizados para induzir a fibrose do músculo gastrocnêmio, e foram tratados com OVAz. A qualidade do colágeno foi avaliada com histologia e à expressão do gene caveolina-1 (CAV-1) foi avaliada usando qPCR. A análise histomorfológica indicou uma redução significativa no perímetro, largura e intensidade do colágeno nos grupos tratados. Os resultados dos níveis de expressão sugeriram diminuição nos grupos de lesão e em dois grupos de tratamento (0,0115 µg/g e 0,009 µg/g). No entanto, com a menor concentração (0,0065 µg/g), não foi observada diferença significativa, apresentando níveis semelhantes aos encontrados em tecido saudável. O uso do OVAz foi eficaz para reverter as alterações do colágeno causadas pela fibrose, e sua menor concentração apresentou uma possível tendência de aumento na expressão do CAV-1. Portanto, os resultados mostraram que o OVAz tem potencial para ser uma alternativa não invasiva e de baixo custo para auxiliar no tratamento da fibrose muscular.

The branched chain amino acids (BCAAs) modulate the development of the intra-cellular stages of Trypanosoma cruzi.

The life cycle of Trypanosoma cruzi, the etiological agent of Chagas disease, involves different forms of the parasite, which alternates between insect and vertebrate hosts. One critical process in the parasite's life cycle is metacyclogenesis, in which the replicative non-infective forms present in the insect midgut differentiate into non-dividing vertebrate-infective forms. It is known that proline (Pro) is important for this process and that leucine (Leu) and isoleucine (Ile) can act as inhibitors of metacyclogenesis. In this study, we investigated further the role of branched-chain amino acids (BCAAs) as negative modulators of parasite differentiation and infection capability in vitro. We found that BCAAs can down-regulate metacyclogenesis, inhibiting Pro-dependent differentiation. Furthermore, we evaluated the ability of all three BCAAs to influence the differentiation of intracellular stages and found that they could modulate the release of trypomastigotes from infected host cells. These findings suggest that BCAAs may have an important role in the complex life cycle of T. cruzi. Thus, enzymes of their metabolism and other interacting proteins could be potential targets for the development of new therapeutic strategies for Chagas disease.

Chemical characterization and effects of volatile oil of Alpinia zerumbet on the quality of collagen deposition and caveolin-1 expression in a muscular fibrosis murine model.

This study evaluated the effect of the volatile oil of Alpinia zerumbet (VOAz) on caveolin-1 gene expression and muscular fibrosis. The rats were immobilized to induce fibrosis of the gastrocnemius muscle, and they were treated with VOAz. Collagen quality was assessed by histology and the expression of the caveolin-1 (CAV-1) gene was evaluated using qPCR. Histomorphological analysis indicated a significant reduction in the perimeter, width, and intensity of collagen in the treated groups, thus showing that the oil was effective in regulating the quality of collagen at the three concentrations. The results of expression levels suggested a decrease in the lesioned group and in two treatment groups (0.0115 µg/g and 0.009 µg/g). However, with the lowest concentration (0.0065 µg/g), no significant difference was observed, with levels similar to those found in healthy tissue. Therefore, the results showed that VOAz has the potential to be a non-invasive and low-cost alternative to aid in the treatment of muscular fibrosis.

Carnauba (Copernicia prunifera) straw as an alternative bedding material for dairy cows housed in a Compost barn system / [Bagana de carnaúba como material de cama alternativo para vacas leiteiras alojadas em sistema Compost barn]

The replacement of bedding in compost dairy barns (CB) comprises a recurrent management practice, but bedding materials are often not readily available in all regions and the choice of alternative materials is necessary. The objective was to evaluate the thermal attributes of carnauba straw (CS) bedding in compost dairy barn facilities. Environmental monitoring operations were performed at a commercial farm located in Northeast Brazil. Mini weather stations were used to evaluate environmental variables. The THI was evaluated as one of the comfort parameters. Analysis of the spatial distribution of bed surface temperature (BST) in the CB was performed using geostatistical techniques. The cows remained out of the comfort zone according to THI results. The BST indicated satisfactory performance and from the thermal point of view can be used as alternative bedding material in CB facilities. However, it was observed that the CS showed fast biomass degradation compared to conventional materials, widely known. In addition, inadequate temperature values (< 45°C) were found in the deeper of the CS bed, signaling higher risks of pathogenic microbial activity. Additional studies are needed for searching the proper management plans that increase the life span of the bed formed by carnauba straw.(AU)

A reposição de cama na instalação Compost Barn (CB) compreende uma prática recorrente de manejo, porém os materiais de cama muitas vezes não estão facilmente disponíveis em todas as regiões, sendo necessária a adoção de materiais alternativos. Objetivou-se avaliar os atributos térmicos da cama à base de bagana de carnaúba (BC) para instalações CB. Foram realizadas operações de monitoramento ambiental em uma fazenda comercial situada no nordeste do Brasil. Miniestações meteorológicas foram utilizadas para avaliação de variáveis ambientais. O ITU foi avaliado como um dos parâmetros de conforto. A distribuição espacial da temperatura superficial da cama (TSC) no galpão foi analisada usando técnicas da geoestatística. As vacas mantiveram-se fora da zona de conforto, conforme resultados do ITU. A TSC indicou desempenho satisfatório e, do ponto de vista térmico, pode ser utilizada como material alternativo para cama em instalações CB. Entretanto, observou-se que a BC apresentou rápida degradação da biomassa em comparação a materiais convencionais, amplamente conhecidos. Além disso, valores inadequados de temperatura (< 45°C) foram encontrados nas camadas profundas da cama de BC, sinalizando maiores riscos de atividade microbiana patogênica. Estudos adicionais tornam-se necessários para a busca de planos de manejo que aumentem o tempo de vida útil da cama formada por bagana de carnaúba.(AU)

Invited review: Compost-bedded pack barns for dairy cows.

Compost-bedded pack barns (CBP) are receiving increasing attention as a housing system for dairy cows that has potential to improve animal welfare. This article reviews current scientific knowledge about CBP with the aim of providing a comprehensive tool for producers and researchers using this housing system. In CBP, cows are provided with an open bedded pack area rather than the individual stalls and concrete alleys found in freestall systems. The bedded pack, a mixture of organic bedding and cattle excreta, is cultivated frequently (1-3 times per day) to incorporate fresh manure and air into the pack, thus promoting an aerobic composting process. To function well, CBP generally require a large area per cow. Optimal animal densities over the bedded area range from 7.4 to more than 15 m2/cow depending on several factors, including climate, bedding, pack management, and cow characteristics. Studies have indicated that CBP, compared with conventional systems such as freestall barns, have the potential to improve the welfare of dairy cows. In particular, the main reported benefits include improved comfort during resting, better foot and leg health, and more natural animal behavior. Research has also indicated that adequate udder health can be achieved in CBP. However, because the bedded pack has been shown to contain high bacterial concentrations, proper management is essential to maintain adequate cow cleanliness and reduce the risk of mastitis. Controlling pack moisture is consistently indicated as the most important issue with CBP. Especially under cold and humid weather conditions, large amounts of bedding may be necessary to keep the pack adequately dry and comfortable for the cows. Nevertheless, the improvements in cow health may offset the higher costs of bedding.

The active transport of histidine and its role in ATP production in Trypanosoma cruzi.

Trypanosoma cruzi, the aetiological agent of Chagas's disease, metabolizes glucose, and after its exhaustion, degrades amino acids as energy source. Here, we investigate histidine uptake and its participation in energy metabolism. No putative genes for the histidine biosynthetic pathway have been identified in genome databases of T. cruzi, suggesting that its uptake from extracellular medium is a requirement for the viability of the parasite. From this assumption, we characterized the uptake of histidine in T. cruzi, showing that this amino acid is incorporated through a single and saturable active system. We also show that histidine can be completely oxidised to CO2. This finding, together with the fact that genes encoding the putative enzymes for the histidine - glutamate degradation pathway were annotated, led us to infer its participation in the energy metabolism of the parasite. Here, we show that His is capable of restoring cell viability after long-term starvation. We confirm that as an energy source, His provides electrons to the electron transport chain, maintaining mitochondrial inner membrane potential and O2 consumption in a very efficient manner. Additionally, ATP biosynthesis from oxidative phosphorylation was found when His was the only oxidisable metabolite present, showing that this amino acid is involved in bioenergetics and parasite persistence within its invertebrate host.

Assessing a mixture of biosurfactant and enzyme pools in the anaerobic biological treatment of wastewater with a high-fat content.

The use of rhamnolipid-type biosurfactant produced by Pseudomonas aeruginosa was evaluated for solubilization of fat present in effluent from a poultry processing plant, followed by anaerobic biological treatment. The rhamnolipid was used in combination with enzyme pools produced by solid medium fermentation of the fungi Penicillium simplicissimum and Penicillium brevicompactum. In experiments with raw effluent, the accumulation of fat caused the specific methane production (SMP) to be much less than with pretreated effluent (0.074 vs. 0.167 L CH4/g chemical oxygen demand (COD) removed). In experiments with pretreated effluent, the SMP ranged from 0.105 to 0.207 L CH4/g CODremoved. A statistical analysis of the results of four sequential batches found that all variables had a significant effect on the SMP in the fourth batch. A fifth batch was initiated for three conditions, and it led to the highest SMP when compared with the control, which showed similar behaviour for the SMP over time, ending up with values three times greater than the SMP in the control conditions. The residual oil and grease analysis revealed removals from 51% to 90% with pretreated effluent and only 1% in the control conditions with raw effluent. Thus, the best synergistic effect of fat release/hydrolysis of effluent components from a poultry processing plant was found with a 0.5% P. brevicompactum pool and rhamnolipid at half the critical micelle concentration (24 mg/L).

The relationship between compost bedded pack performance, management, and bacterial counts.

The objective of this study was to assess the relationships among temperature, moisture, carbon-to-nitrogen (C:N) ratio, space per cow, and bacterial counts from bedding material collected from compost bedded pack (CBP) barns. A field survey of 42 routinely aerated CBP barns was conducted in Kentucky between October 2010 and March 2011. Two bedding material samples of 1,064.7 cm(3) each were collected during a single site visit from 9 evenly distributed locations throughout each barn and thoroughly mixed to create a composite sample representative of the entire CBP. Bacterial counts were determined for coliforms, Escherichia coli, streptococci, staphylococci, and Bacillus spp. University of Kentucky Regulatory Services (Lexington) laboratory personnel performed nutrient analyses to determine moisture, carbon, and nitrogen contents. Surface and 10.2-cm pack depth temperatures were collected for each of the 9 evenly distributed locations and the mean calculated to produce a composite temperature. Space per cow was calculated as the total CBP area divided by number of cows housed on the CBP. The GLM procedure of SAS (SAS Institute Inc., Cary, NC) generated models to describe factors affecting bacterial counts. Bacterial counts were 6.3 ± 0.6, 6.0 ± 0.6, 7.2 ± 0.7, 7.9 ± 0.5, and 7.6 ± 0.5 log 10 cfu/g of dry matter for coliform, Escherichia coli, streptococci, staphylococci, and Bacillus spp., respectively. Composite temperature, CBP moisture, C:N ratio, and space per cow had no effect on coliform counts. Escherichia coli reached a peak concentration when the C:N ratio was between 30:1 and 35:1. Staphylococci counts increased as ambient temperature increased. Streptococci counts decreased with increased space per cow and composite temperature and increased with increasing ambient temperature and moisture. Streptococci counts peaked at a C:N ratio ranging from 16:1 to 18:1. Bacillus spp. counts were reduced with increasing moisture, C:N ratio, and ambient temperature. Mastitis-causing bacteria thrive in similar conditions to that of composting bacteria and microbes, making elimination of these at higher temperatures (55 to 65°C) difficult in an active composting environment. Producers must use recommended milking procedures and other preventative practices to maintain low somatic cell count in herds with a CBP barn.

Cytotoxic mechanism of Piper gaudichaudianum Kunth essential oil and its major compound nerolidol.

Piper gaudichaudianum Kunth is used in popular medicine as anti-inflamatory and against liver disorders. One of the most studied components of the plant is the essential oil for which chemical analysis revealed (E)-nerolidol as major compound. Recently, we have shown that P. gaudichaudianum essential oil possesses strong cytotoxic effects in mammalian V79 cells. The aim of this study was to analyze the cytotoxicity and mutagenicity of P. gaudichaudianum essential oil and nerolidol using Saccharomyces cerevisiae as model study. Treatment of the XV185-14c and N123 strains with essential oil and nerolidol led to cytotoxicity but did not induce mutagenicity. Our results revealed an important role of base excision repair (BER) as the ntg1, ntg2, apn1 and apn2 mutants showed pronounced sensitivity to essential oil and nerolidol. In the absence of superoxide dismutase (in sod1Δ mutant strain) sensitivity to the essential oil and nerolidol increased indicating that this oil and nerolidol are generating reactive oxygen species (ROS). The ROS production was confirmed by DCF-DA probing assay in Sod-deficient strains. From this, we conclude that the observed cytotoxicity to P. gaudichaudianum essential oil and nerolidol is mainly related to ROS and DNA single strand breaks generated by the presence of oxidative lesions.

Compost bedded pack dairy barn management, performance, and producer satisfaction.

The objective of the research was to characterize herd performance, producer satisfaction and recommendations, and management practices used by compost bedded pack (CBP) managers in Kentucky (42 farms and 47 CBP facilities). Farms were visited between October 2010 and March 2011. A random selection of cows housed solely in the CBP were scored for locomotion and hygiene. Changes in monthly Dairy Herd Improvement Association performance records, including milk production, SCC, reproductive performance, and daily bulk-tank somatic cell count after moving into the CBP were analyzed using the MIXED procedure of SAS (SAS 9.3; SAS Institute Inc., Cary, NC). The GLM procedure of SAS (SAS 9.3) was used to develop models to describe CBP moisture, CBP temperature at 20.3 cm, and mean herd hygiene. Producers provided 9.0 ± 2.2 m2 of pack space per cow (n = 44). Barns constructed with an attached feed alley cost $1,051 ± 407 per cow (n = 40). Barns constructed without an attached feed alley cost $493 ± 196 per cow (n = 13). Kiln-dried shavings required 0.05 ± 0.04 m3 of bedding per cow per day (n = 15). Green shavings required 0.07 ± 0.06 m3 of bedding per cow per day (n = 12). The most-frequently cited benefits of the CBP included cow comfort (n = 28), cow cleanliness (n = 14), and the low-maintenance nature of the system (n = 10). Increased stirring frequency, stirring depth, and ambient temperature increased pack temperature, measured at 20.3 cm below the CBP surface. Increased stirring depth, pasture-adjusted space per cow, and drying rate decreased CBP moisture. Mean herd locomotion and hygiene scores were 1.5 ± 0.3 (n = 34) and 2.2 ± 0.4 (n = 34), respectively. Increased 20.3-cm depth CBP temperature and ambient temperatures improved mean herd hygiene. Bulk-tank somatic cell count decreased from the year before to the year after moving into the CBP barn (323,692 ± 7,301 vs. 252,859 ± 7,112 cells/mL, respectively) for farms using the CBP barn as the primary housing facility (n = 9). Daily milk production, collected from monthly Dairy Herd Improvement Association tests, increased from before moving into the CBP barn to the second year after (29.3 ± 0.3 vs. 30.7 ± 0.3 kg, respectively) for farms using the CBP barn as the primary housing facility (n = 8). Calving interval decreased from the year before to the second year after (14.3 ± 0.1 vs. 13.7 ± 0.1 mo) moving into the CBP barn for farms using the CBP as primary housing (n = 8).

Chemical composition and cytotoxic, mutagenic and genotoxic activities of the essential oil from Pipergaudichaudianum Kunth leaves.

We have investigated the chemical composition of Piper gaudichaudianum essential oil, as well as its cytotoxic, mutagenic and genotoxic effects in V79 cells. The chemical analyses showed that the major compounds are (E)-nerolidol (22.4%), alpha-humulene (16.5%), (E)-caryophyllene (8.9%) and bicyclogermacrene (7.4%). Dose-dependent cytotoxic effects were observed in V79 cells treated with essential oil by using clonal survival, 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide reduction assay (MTT) and trypan blue exclusion assay (TB), and a significant decrease in survival was observed at concentrations of 0.5 microg/mL and higher. The P. gaudichaudianum essential oil treatment caused DNA strand breaks in V79 cells at concentrations up to 2 microg/mL, as detected by the alkaline comet assay, but did not induce double-strand breaks, as verified by neutral comet assay. It induced a significant increase in the frequency of micronucleated cells at 4, 6 and 10 microg/mL. Moreover, P. gaudichaudianum essential oil significantly increased lipid peroxidation at doses of 0.5 microg/mL and higher, suggesting that the observed oxidant potential can be responsible, at least in part, for its cytotoxic and genotoxic effects.

[Sodoku--a case report]. / Sodoku--descrição de um caso.

Rat-bite fever results from an infection with the organism Spirillum minus. The authors report a symptomatic patient with fever, malaise, and ulcer in forehead after a rodent exposure. The organism was identified in darkfield examination of the ulcer exudate. Penicillin was the drug used with clinical improvement of symptoms.