Metabolomics approach for phenolic compounds profiling of soursop (Annona muricata L.) fruit during postharvest storage.

INTRODUCTION: Soursop (Annona muricata L.) is a crop with medicinal properties and numerous bioactive compounds. Ripening is a complex process that regulates fruit quality and changes in metabolite content, such as flavonoids, polyphenols, and organic acids. OBJECTIVES: This study aimed to analyze the phenolic profiling of soursop fruit ripening. METHODS: The metabolic changes in different days of storage of soursop fruits were investigated using a semi-metabolomic approach based on ultra-performance liquid chromatography coupled to electrospray ionization quadrupole-time of flight mass spectrometry (UPLC-ESI-QTOF-MS). Further, multivariate analysis such as supervised partial least squares discriminant analysis (PLS-DA) was conducted to identify differential metabolites. RESULTS: A total of 68 metabolites were identified in soursop fruit during postharvest storage. A higher number of metabolites were identified in the Day zero (D0) compared to the Day one (D1), Day three (D3), and Day five (D5), belonging to flavonoids, other polyphenols, phenolic acids, and organic acids. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the pathways of flavone and flavonol biosynthesis, flavonoid biosynthesis, and biosynthesis of secondary metabolites were mostly enriched. Additionally, we included all the compounds and their postharvest storage in the public Phenolics profile database. CONCLUSIONS: Here, we show that the stage of ripening has a significant effect on the phenolic content, highlighting the point of cut (D0) and the onset of senescence (D5). The findings of this study provide new insights into the soursop fruit quality and may contribute to the identification of metabolic markers for its storage.

Bacillus mojavensis enhances the antioxidant defense mechanism of soursop (Annona muricata L.) fruits during postharvest storage.

Rapid softening of soursop (Annona muricata L.) fruit results in postharvest losses. Bacillus genus is one of the most studied antagonistic biological control agents against postharvest diseases. Nevertheless, information about how this bacterium acts on the fruits is still not understood. The objective of this study aims to gain an insight into the effect of Bacillus mojavensis on the activity and gene expression of antioxidant defense enzymes in soursop fruits during postharvest storage. Our findings indicate different responses in the fruits inoculated with B. mojavensis at biochemical and molecular levels. On day one, fruits inoculated with B. mojavensis presented a mean value of 79.09 GAE/100 gFW in total phenols, and higher superoxide dismutase (SOD) and catalase (CAT) activities (1.35 and 1.78-fold higher, respectively). On the other hand, on the third day of storage, the ferric reducing/antioxidant power (FRAP) reached its highest level, including an increase in the expression of SOD, and PPO genes by 18.7-fold and 4.5-fold in fruits inoculated with B. mojavensis. Finally, on the fifth day of storage, soursop fruits inoculated with B. mojavensis had the highest mean values for 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH·), 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonate (ABTS· +), with values of 194.68 EAA/100 gFW, and 172.33 EAA/100 gFW, respectively. Indeed, higher polyphenol oxidase (PPO), and peroxidase (POD) activities (2.17-fold and 1.27-fold higher, respectively) were recorded compared to the control fruits. We show that depending on the stage of ripening, the antagonist bacteria B. mojavensis enhanced the antioxidant capacity, enzymatic activity, and gene expression of soursop fruits.

Salpianthus macrodontus Extracts, a Novel Source of Phenolic Compounds with Antibacterial Activity against Potentially Pathogenic Bacteria Isolated from White Shrimp.

This study aimed to evaluate the antibacterial activity in vitro of Salpianthus macrodontus and Azadirachta indica extracts against potentially pathogenic bacteria for Pacific white shrimp. Furthermore, the extracts with higher inhibitory activity were analyzed to identify compounds responsible for bacterial inhibition and evaluate their effect on motility and biofilm formation. S. macrodontus and A. indica extracts were prepared using methanol, acetone, and hexane by ultrasound. The minimum inhibitory concentration (MIC) of the extracts was determined against Vibrio parahaemolyticus, V. harveyi, Photobacterium damselae and P. leiognathi. The polyphenol profile of those extracts showing the highest bacterial inhibition were determined. Besides, the bacterial swimming and swarming motility and biofilm formation were determined. The highest inhibitory activity against the four pathogens was found with the acetonic extract of S. macrodontus leaf (MIC of 50 mg/mL for Vibrio spp. and 25 mg/mL for Photobacterium spp.) and the methanol extract of S. macrodontus flower (MIC of 50 mg/mL for all pathogens tested). Both extracts affected the swarming and swimming motility and the biofilm formation of the tested bacteria. The main phenolic compounds related to Vibrio bacteria inhibition were naringin, vanillic acid, and rosmarinic acid, whilst hesperidin, kaempferol pentosyl-rutinoside, and rhamnetin were related to Photobacterium bacteria inhibition.

Growth Promoting Activity of Annona muricata L. Leaf Extracts on Lactobacillus casei.

Soursop leaves are a source of phytochemical compounds, such as phenolic acids, flavonoids, hydrolyzable tannins, and acetogenins. These compounds can have several types of biological activities. Lactic acid bacteria can uptake phenolic compounds present in plants or fruits. The aim of the present work was to investigate the in vitro effect of hexane, acetone, methanolic, and aqueous extracts of soursop leaves (Annona muricata L.) on the growth, motility, and biofilm formation of Lactobacillus casei, and to determine compounds related to growth. The minimum concentration promoting growth, motility (swimming, swarming, and twitching), and biofilm-forming capacity (crystal violet) were evaluated. The results showed the growth-promoting capacity of acetone and aqueous extracts at low doses 25-50 mg/L, and an inhibition in the four extracts at higher doses of 100 mg/L. The L. casei growth is related to ellagic acid, quercetin rhamnoside, kaempferol dihexoside, quercetin hexoside, secoisolariciresinol, and kaempferol hexoside-rhamnoside. Hexane extract increased the three types of motility, while aqueous maintained swimming and twitching motility similar to control. The four extracts inhibited the biofilm formation capacity.

Differential Responses of Antioxidative System during the Interaction of Soursop Fruits (Annona muricata L.) and Nectria haematococca at Postharvest Storage.

Soursop fruit (Annona muricata L.) production is diminished by the attack of pathogens such as Nectria haematococca. However, the fruit-pathogen interaction at the biochemical and molecular levels is still unknown. The objective of this study was to analyze the response of the soursop fruit to the presence of N. haematococca during postharvest storage. Soursop fruits were inoculated with the pathogen and total phenolic compounds, antioxidant capacity by Ferric reducing/antioxidant power (FRAP), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS•+), and 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH•), as well as enzymatic activity and transcript levels of polyphenol oxidase (PPO) and superoxide dismutase (SOD), were evaluated at 1, 3, and 5 days of storage. The noninoculated fruits were the controls of the experiment. The highest total phenol content was recorded on day one in the inoculated fruits. FRAP, ABTS, and DPPH activity presented the highest values on day three in the control fruits. Inoculated fruits recorded the highest PPO activity on day five and a five-fold induction in the PPO transcript on day three. SOD activity showed a decrease during the days of storage and 10-fold induction of SOD transcript on day three in the inoculated fruits. Principal component analysis showed that total phenols were the variable that contributed the most to the observed variations. Furthermore, a positive correlation between total phenols and SOD activity, PPO expression, and SOD expression, as well as between DPPH and FRAP, was recorded. The results showed a differential response in antioxidant capacity, enzymatic activity, and gene expression during the interaction of soursop fruits-N. haematococca at postharvest storage.

Characterization and proliferation capacity of potentially pathogenic fungi in marine and freshwater fish commercial feeds.

In the aquaculture industry, the selection and quality of feed are highly relevant because their integrity and management have an impact on the health and development of organisms. In general, feeds contamination depends on storage conditions and formulation. Furthermore, it has been recognized that filamentous fungi are among the most important contaminating agent in formulated feeds. Therefore, the purpose of this research was to identify saprophytic fungi capable of proliferating in commercial feeds, as well as determining their prevalence, extracellular enzymes profile, ability to assimilate carbon sources, and finally their ability to produce aflatoxins. In order to do that, twenty-two fungi were isolated from commercial fish feeds. After, the species Aspergillus chevalieri, A. cristatus, A. sydowii, A. versicolor, A. flavus, A. creber, and Lichtheimia ramosa were identified. These fungi were able to produce extracellular enzymes, such as phosphatases, esterases, proteases, ß-glucosidase, and N-acetyl-ß-glucosaminidase. The isolated fungi showed no selective behavior in the assimilation of the different carbon sources, showing a strong metabolic diversity. Prevalence percentages above 85% were recorded. Among all fungi studied, A. flavus M3-C1 had the highest production of aflatoxins when this strain was inoculated directly in the feeds (295 ppb). The aflatoxin production by this strain under the experimental setting is above the permitted levels, and it has been established that high levels of aflatoxins in feeds can cause alterations in fish growth as well as the development of cancerous tumors in the liver, in addition to enhancing mortality.

Cold Plasma Treatment as an Alternative for Ochratoxin a Detoxification and Inhibition of Mycotoxigenic Fungi in Roasted Coffee.

Ochratoxin A (OTA) produced by mycotoxigenic fungi (Aspergillus and Penicillium spp.) is an extremely toxic and carcinogenic metabolite. The use of cold plasma to inhibit toxin-producing microorganisms in coffee could be an important alternative to avoid proliferation of mycotoxigenic fungi. Roasted coffee samples were artificially inoculated with A. westerdijikiae, A. steynii, A. versicolor, and A. niger, and incubated at 27 °C over 21 days for OTA production. Samples were cold plasma treated at 30 W input power and 850 V output voltage with helium at 1.5 L/min flow. OTA production in coffee was analyzed by high performance liquid chromatography coupled to a mass spectrometer (HPLC-MS). After 6 min of treatment with cold plasma, fungi were completely inhibited (4 log reduction). Cold plasma reduces 50% of OTA content after 30 min of treatment. Toxicity was estimated for extracts of artificially contaminated roasted coffee samples using the brine shrimp (Artemia salina) lethality assay. Toxicity for untreated roasted coffee was shown to be "toxic", while toxicity for cold plasma treated coffee was reduced to "slightly toxic". These results suggested that cold plasma may be considered as an alternative method for the degradation and reduction of toxin production by mycotoxigenic fungi in the processing of foods and feedstuffs.

Safety and Therapeutic Profile of a GnRH-Based Vaccine Candidate Directed to Prostate Cancer. A 10-Year Follow-Up of Patients Vaccinated With Heberprovac.

Heberprovac is a GnRH based vaccine candidate containing 2.4 mg of the GnRHm1-TT peptide as the main active principle; 245 µg of the very small size proteoliposomes adjuvant (VSSP); and 350 µL of Montanide ISA 51 VG oil adjuvant. The aim of this study was to assess the safety and tolerance of the Heberprovac in advanced prostate cancer patients as well as its capacity to induce anti-GnRH antibodies, the subsequent effects on serum levels of testosterone and PSA and the patient overall survival. The study included eight patients with histologically-proven advanced prostate cancer with indication for hormonal therapy, who received seven intramuscular immunizations with Heberprovac within 18 weeks. Anti-GnRH antibody titers, testosterone and PSA levels, as well as clinical parameters were recorded and evaluated. The vaccine was well tolerated. Significant reductions in serum levels of testosterone and PSA were seen after four immunizations. Castrate levels of testosterone were observed in all patients at the end of the immunization schedule, which remained at the lowest level for at least 20 months. In a 10-year follow-up three out of six patients who completed the entire trial survived. In contrast only one out eight patients survived in the same period in a matched randomly selected group receiving standard anti-hormonal treatment. Heberprovac vaccination showed a good security profile, as well as immunological, biochemical and, most importantly, clinical benefit. The vaccinated group displayed survival advantage compared with the reference group that received standard treatment. These results warrant further clinical trials with Heberprovac involving a larger cohort.

Emprego de células tronco na odontologia / The use of stem cells in dentistry

Objetivo: com este estudo propôs-se realizar uma revisão integrativa da literatura a respeito do uso de célulastroncona odontologia. Buscou-se identificar o atual uso da terapia com células-tronco e suas potencialidades que apesquisa aponta para a terapia com células-tronco adultas e embrionárias, analisando seus limites relacionados aouso em seres humanos. Método: os dados foram coletados mediante revisão da literatura utilizando os descritorescélulas-tronco, odontologia e terapia, tomando como base artigos levantados dos últimos dez anos publicados atravésda Biblioteca Virtual em Saúde (BVS), além de textos clássicos sobre o tema. Como critérios de inclusão foramestabelecidos artigos originais e de revisão publicados na íntegra na forma on-line nos idiomas inglês e portuguêspublicados no período de 2004 a 2014, utilizando os descritores supramencionados nos respectivos idiomas. Foramutilizadas na busca a base de dados Biblioteca Virtual em Saúde. De maneira complementar foram incluídos doiscapítulos de título em formato de livro pela relevância da abordagem na obra referenciada. Como critério de exclusãofoi estabelecido artigos repetitivos nas bases de dados, além de dissertações, teses e editoriais pertinentes à buscanos descritores definidos. Resultados: os resultados mostram que apesar dos grandes estudos realizados, a utilizaçãodas células-tronco na odontologia não se tornou rotina de protocolo terapêutico na magnitude de suas indicações. Ascélulas-tronco embrionárias ainda apresentam grande limitação quando comparadas às células-tronco adultas, estas,por sua vez, tem se mostrado bastante promissoras.

Objective: With this study we propose to accomplish an integrative review of the literature regarding the use of stemcells in dentistry. The study attempted to identify the current use of the stem cell therapy and its potential which is ledby the research to the therapy with adult and embryonic stem cells through the analysis of its limits related to the usein humans and animals. Methods: The data was collected through a literature review using the descriptors stem cells,dentistry and therapy, based on the survey of articles published in the last five years through the Virtual Health Library,and also classic texts about the topic. The inclusion criterion were established original and review articles publishedin full in the online form in English and Portuguese published from 2004 to 2014, using the above descriptors in theirrespective languages were used to search the database Virtual Health Library. In a complementary way were includedtwo title chapters in book form by the relevance of the approach in the referenced work. The exclusion criterion wasestablished repetitive articles in the databases, as well as dissertations, theses and editorials relevant to search thedefined descriptors. Results: the results show that despite the major studies, the use of stem cells in dentistry didnot become routine therapeutic Protocol in the importance/magnitude of their indications. Embryonic stem cells stillshow great rejection when compared with adult stem cells.

Factores de riesgo y manejo de la infección en la artroplastía total de rodilla / Risk factors and treatment for infection in total knee replacement

Objetivo. Con el presente estudio se trata de establecer los factores de riesgo para la artroplastía total de la rodilla, su frecuencia de infección y la eficacia del tratamiento para las complicaciones. Material y métodos. Estudio retrospectivo extraído de un total de 447 casos de prótesis total de rodilla (PTR) operados en un periodo de 10 años. Se diseñaron dos grupos comparativos, uno con 19 casos infectados y otro con 237 no infectados, tomados al azar, para un total de 256, con edad promedio de 68 años. Resultados. La edad promedio en mujeres con infección fue de 71 años (no infectadas 60 años, p = 0.002). Para mujeres infectadas con AR fue de 78 años (no infectadas con AR 54 años, p = 0.0001). No hubo diferencia entre mujeres que tenían enfermedades coexistentes, independientemente de artrosis o AR. En hombres con infección, la edad promedio fue 56.9 años para AR y 72.6 para artrosis (p = 0.0001). En todos los casos se resolvió mediante diversos procedimientos, como recambio del componente rotuliano en uno, recambio total en dos, desbridamiento y colgajos en cinco y artrodesis en 10. Conclusión. Los principales riesgos de infección parecen ser, la existencia en la 8a década de la vida para mujeres, independientemente de sufrir o no AR así como la presencia de AR en hombres menos viejos. La sola edad en varones, y la artrosis o las enfermedades coexistentes indistintamente del sexo no parecen ser por sí solas, factores de riesgo significativo.

Eficacia comparativa entre los abordajes lateral y medial para corregir la deformidad en valgo mayor de 15 grados en la artroplastía total de la rodilla / Comparative trial betwenn medial and lateral approach for corrrecting valgus of the knee, greater than 15 degrees in total knee replacement

Objetivo. Valorar la conveniencia de utilizar el abordaje pararrotuliano lateral en la artoplastía total de la rodilla (ATR) en pacientes con valgo exagerado de la rodilla. Material y métodos. Se trata de un estudio retrospectivo de 13 mujeres (de un total de 14), operadas de ATR de 1995 a 2000, con un valgo preoperatorio mayor de 15 grados. Sin selección al azar, el abordaje lateral se usó en 6 (prom. 60.8 años y valgo 17.5º) y el medial en 7 (70.4 años, valgo 17.2º). La causa fue artrosis en los seis de abordaje medial y en cinco de los siete con abordaje lateral, y AR en los dos restantes. Resultados. Con base en los parámetros de dolor postoperatorio, estabilidad articular y arco de movimiento, los resultados fueron buenos en cuatro y malos en tres para el abordaje medial, mientras que fueron buenos en cinco y malos en uno para el abordaje lateral. El único resultado más notorio es la diferencia entre los resultados para el abordaje lateral, que parece orientarse a su favor, ya que en el caso del abordaje medial no hay diferencia alguna. Conclusión. El abordaje pararrotuliano lateral debe considerarse como una alternativa para la ATR en virtud de la tendencia aparente a cursar con menor proporción de complicaciones en pacientes con genu valgo exagerado.

Artroplastía total de rodilla sin componente rotuliano / Total knee replacement with preservation of the patient's patella

Se estudiaron 83 pacientes a los que se les realizó un total de 89 artroplastías de rodilla, evaluándose las ventajas y desventajas de no realizar la colocación del implante rotuliano. Solamente se incluyeron en el estudio pacientes con diagnóstico de gonartrosis degenerativa. Los pacientes se evaluaron con el sistema propuesto por the Knee Society antes y después de la cirugía, agregándose las complicaciones rotulianas como son luxación, subluxación y fractura patelar. Los resultados demostraron fractura de rótula en cinco pacientes (5.61 por ciento), dolor rotuliano en 15 (16.85 por ciento), y rigidez de rodilla en cinco (5.61 por ciento). No se presentaron casos de luxación o subluxación rotuliana. Al no colocar el botón rotuliano se disminuyen aún más las posibles complicaciones propias del implante en forma significativa

Valoración del incremento acetabular después de osteotomía innominada del iliaco: modelo experimental / An assessment of the acetabular increase after the innominate osteotomy of the iliac bone: experimental model

En 1915 Albee inició el tratamiento de la luxación congénita de la cadera comparando sus resultados con los reportados por Hoffa en 1890. A partir de este momento se inició una serie de publicaciones en la literatura anglosajona sobre osteotomías del iliaco. La forma en que describen el desplazamiento acetabular es con base en una evaluación radiográfica que sólo es bidimensional, y clínicamente en cadáver o paciente, en forma tridimencional, pero de manera subjetiva. En el año de 1976, George T. Rab describió los aspectos biomecánicos de la osteotomía de Salter; posteriormente, en 1978 Hansson reportó un caso clínico con valoración tridimensional con el uso de la radioesterofotogrametría (REFG) del desplazamiento acetabular después de la osteotomía del iliaco. El porcentaje de error en ambas técnicas es alto en forma cuantitativa, pero no en forma cualitativa. En esta investigación empleamos el método de la REFG, para determinar el desplazamiento acetabular en forma tridimensional, que consta de una jaula de calibración, marcadores radiopacos, dos placas radiográficas y, por último una pelvis, modelo de acetábulos displásicos tomado del catálogo de Sawbones. Los resultados fueron un desplazamiento del acetábulo después de la osteotomía innominada del iliaco en forma anterolateral y con desplazamiento medial de la pared posterior, lo cual condiciona una falta de cobertura posterior. El centro del acetábulo se desplaza en forma dista, posterior y medial. Concluimos que el método de REFG es eficaz para la valoración del desplazamiento acetabular en forma tridimencional, en el modelo experimental, tanto en forma cuantitativa como cualitativa.

Instrumentación segmentaria de la columna en la distrofia muscular progresiva / Segmental instrumntation of spine in progressive muscular distrophy. Report of 15 patients

Se presenta una serie de 15 pacientes con distrofia muscular progresiva (DMP) que fueron instrumentados en el Hospital Shiriners para Niños Lisiados de la Ciudad de México, de 1976 a 1990. El criterio para la cirugía se basó en la disminución de la capacidad vital a un mínimo de 30%, a la progresión de la deformidad de la columna en pacientes no ambulatorios, o a ambas cosas. El seguimiento fue de seis años como promedio y la medición de las curvas xifoescolióticas fue de 26.5 grados en promedio. La edad al momento de la cirugía fue entre cinco y 14 años. En seis pacientes se instrumentó de C2 a S1 para obtener control de la cabeza, ocho pacientes de C5 a S1 y un paciente de T1 a S1. El resultado fue la estabilidad y el equilibrio cifolordótico de la columna, la mejoría en la capacidad vital, la disminución de los cuadros infecciosos respiratorios y principalmente una mejor calida de vida de los pacientes.