Advances in analytical techniques coupled to in vitro bioassays in the search for new peptides with functional activity in effect-directed analysis.

Bioactive peptides derived from food proteins have important biological effects. The search for new bioactive peptides from natural sources needs to base their decisions on evidence from methods providing structural information, where classic biological assays do not provide it. In this context, high-performance thin-layer chromatography (HPTLC) coupled with bioassays in effect-directed analysis (EDA) has been used recently to detect the bioactivities of peptides. In EDA, peptides are separated and the biological effects of individual fractions can be determined. HPTLC-EDA coupled with MS represents an emerging technology, complementary to the more common HPLC and electrophoresis analytical methods used for peptide analysis. In addition, thin-layer chromatography (TLC) can be used to identify the origins of substances causing biological effects. This review provides an overview of the status of analytical methods in these systems as well as new strategies for the detection and characterization of bioactive peptides.

Phycobiliproteins: Structural aspects, functional characteristics, and biotechnological perspectives.

Phycobiliproteins (PBPs) are fluorescent proteins of various colors, including fuchsia, purple-blue and cyan, that allow the capture of light energy in auxiliary photosynthetic complexes called phycobilisomes (PBS). PBPs have several highly preserved structural and physicochemical characteristics. In the PBS context, PBPs function is capture luminous energy in the 450-650 nm range and delivers it to photosystems allowing photosynthesis take place. Besides the energy harvesting function, PBPs also have shown to have multiple biological activities, including antioxidant, antibacterial and antitumours, making them an interesting focus for different biotechnological applications in areas like biomedicine, bioenergy and scientific research. Nowadays, the main sources of PBPs are cyanobacteria and micro and macro algae from the phylum Rhodophyta. Due to the diverse biological activities of PBPs, they have attracted the attention of different industries, such as food, biomedical and cosmetics. This is why a large number of patents related to the production, extraction, purification of PBPs and their application as cosmetics, biopharmaceuticals or diagnostic applications have been generated, looking less ecological impact in the natural prairies of macroalgae and less culture time or higher productivity in cyanobacteria to satisfy the markets and applications that require high amounts of these molecules. In this review, we summarize the main structural characteristics of PBPs, their biosynthesys and biotechnological applications. We also address current trends and future perspectives of the PBPs market.

Chemometric optimization of trypsin digestion method applying infrared, microwave and ultrasound energies for determination of caseins and ovalbumin in wines.

Caseins and ovalbumin are frequently used as wine fining agents to remove undesirable compounds like polymeric phenols. Their presence in wines is a subject of concern because may cause adverse effects on susceptible consumers, especially when their presence is not labeled. A key step for its determination is trypsin digestion, which is considered the bottleneck of bottom-up approach workflow because usually requires several hours. To reduce this time, the objective of this work was to carry out a chemometric optimization of trypsin digestion method applying infrared, microwave and ultrasound energies to determine caseins and ovalbumin in wines. The conditions of each accelerated digestion method were optimized using a Response Surface Methodology based on central composite design. The parameters optimized were digestion time and trypsin: protein ratio. The response variable evaluated was digestion yield, which was determined through the peak area of each protein transition determined by liquid chromatography-mass spectrometry. The most effective technique was microwave followed by ultrasound and infrared. Since optimal values of microwave and ultrasound-assisted digestion were the same, the later was chosen considering sample preparation and cost. Applying the proposed approach, a reduction of ca. 140 and 240-fold on digestion time was achieved compared with optimized and non-optimized conventional methods, respectively. With this workflow, both proteins were digested in a single 3 min process allowing its detection by liquid chromatography-mass spectrometry at µg L-1 level, which is ca. 60 times lower than the current limit of 0.25 mg L-1.

Identification of type B trichothecenes and zearalenone in Chilean cereals by planar chromatography coupled to mass spectroscopy.

High-performance thin-layer chromatography (HPTLC) and HPTLC coupled with mass spectrometry (MS) methods were described for the simultaneous determination of zearalenone (ZEA); type B trichothecenes (TCT-B); nivalenol (NIV) and deoxynivalenol (DON) along with its acetylated derivatives: 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON). The extract samples were cleaned-up with Bond Elut Mycotoxin® solid-phase extraction cartridges. Then, separation was performed on HPTLC silica gel 60 F254 plates using toluene, ethyl acetate and formic acid (1:8:1 v/v/v) as mobile phase. Derivatisation was then performed with 10% aluminium trichloride in 50% methanol. Mycotoxin standards and spiked cereals grains were identified by UV spots at 366 nm, with retention factors (RF) of 0.20 (NIV), 0.39 (DON), 0.45 (15-ADON), 0.50 (3-ADON) and 0.60 (ZEA). Some parameters of validation were determined. Calibration data (n = 5) fitted a linear regression model with determination coefficients, R2 > 0.99. The recovery was determined in triplicate at two levels, ranging from 84.3 ± 2.2% to 114.2 ± 11.7%. Detection limits ranged from 80 to 120 µg kg-1 and quantification limits ranged from 120.0 to 200 µg kg-1. The analysis by HPTLC/electrospray (ESI)-MS in negative mode confirmed the presence of TCT-B and ZEA standards in Chilean cereals with mass signals at m/z 355, 371, 337, and 317 for DON, NIV, 3-ADON and 15-ADON, and ZEA, respectively.

Occurrence of allergen proteins in wines from Chilean market.

Casein and ovalbumin are proteins commonly used as wine fining agents that may trigger allergic reactions in susceptible individuals. Therefore, their occurrence in wines could become a health risk, moreover when their presence is not reported. The objective of this work was to determine the occurrence of casein and ovalbumin in Chilean wines. Proteins were extracted by ultrafiltration and digested applying an optimised ultrasound-assisted method. Peptides were quantified by validated tandem mass spectrometry method using stable isotope dilution analysis. Optimal digestion was achieved in 3 minutes at a 1:10 enzyme protein ratio. The method was validated following ICH guidelines showing determination coefficients R2 ≥ 0.99, repeatability, and intermediate precision with RSD values <1.95% and recoveries from 89.8% to 115.1% (RSD < 5.84%). The method was applied to analyse 60 wine samples. Fifty-six samples showed quantifiable levels, from which 28% presented a total casein and ovalbumin concentration equal or higher to the European limit (0.25 mg L-1).

An improved method for a fast screening of α-glucosidase inhibitors in cherimoya fruit (Annona cherimola Mill.) applying effect-directed analysis via high-performance thin-layer chromatography-bioassay-mass spectrometry.

α-glucosidase inhibitors (AGIs) are very attractive bioactive compounds due to their therapeutic profile that includes beneficial effects over glycemic control in type 2 diabetes mellitus and viral infections. Its detection and identification in plants and fruits has gained growing attention, and certainly requires efficient screening methodologies. The objective of the present work was to develop a fast methodology to detect and identify AGIs in cherimoya fruit (Annona cherimola Mill.) applying effect-directed analysis via high-performance thin layer-chromatography (HPTLC) linked with bioassay and mass spectrometry (MS). Both, HPTLC and bioassay conditions, were optimized accomplishing 50% and 83% reduction on enzyme concentration and incubation time respectively, compared to the original method. Additionally, the contrast between inhibitory bands and purple background was also enhanced by enzyme substrate impregnation on HPTLC plate. The optimized detection conditions established were the following: 5.0 U mL-1 of enzyme solution, 1.0 mg mL-1 of 2-naphthyl-α-D-glucopyranoside substrate, 1.0 mg mL-1 of Fast Blue B salt solution and 10 min as incubation time. Applying this methodology, coupled to HPTLC-MS and ultra-high-performance liquid chromatography (UHPLC)-diode array detector (DAD)-MS/MS, it was possible for the first time to detect and identify three AGIs in cherimoya peel and seeds. Compounds were tentatively assigned as phenolamides (phenylethyl cinnamides): N-trans-feruloyl tyramine (m/z 314 [M+H]+; UV λmax 293 and 316 nm), N-trans-p-coumaroyl tyramine (m/z 284 [M+H]+; UV λmax 296 nm) and N-trans-feruloyl phenethylamine (m/z 298 [M+H]+; UV λmax 288 nm). To the best of our knowledge, the presence of latter compound is reported for the first time in cherimoya.

Utilización de antimicrobianos en el Hogar materno Doña Leonor Pérez durante el período 2007-2009 / Use of antimicrobials in Doña Leonor Pérez home for pregnant women in the 2007-2009 period

Introducción: el uso inadecuado de los medicamentos supone un importante problema sanitario y económico, más sensible en el caso de los niños, ancianos y las embarazadas. Existen una gran diversidad de medicamentos y sobre muchos de ellos no se tiene un criterio definitivo de su inocuidad, por lo que la información disponible para la prescripción de fármacos en mujeres embarazadas, es muy escasa. Objetivo: evaluar la calidad en el uso racional de antimicrobianos en las gestantes ingresadas en el Hogar materno Doña Leonor Pérez Cabrera durante el período 2007- 2009. Métodos: este trabajo constituye un estudio de utilización de medicamento del tipo indicación prescripción, retrospectivo y descriptivo. Se revisaron las historias clínicas y se obtuvieron las variables biosociales edad materna, edad gestacional al ingreso, ocupación, diagnóstico que precisó el uso de antimicrobianos, forma de diagnóstico más utilizado así como los antimicrobianos más prescritos. Resultados: a través del análisis de las variables y de los esquemas terapéuticos se pudo evaluar la calidad de la prescripción de los antimicrobianos. Los resultados obtenidos permitieron constatar que predominaron las gestantes con edades entre 21 y 30 años, con edades gestacionales al ingreso entre 21 y 30 semanas y trabajadoras. La patología que mayormente motivó la utilización de antimicrobianos fue la sepsis vaginal, diagnosticada de forma clínica en casi la totalidad de los casos y los antimicrobianos más prescritos fueron la azitromicina, metronidazol, clotrimazol y nistatina. Conclusiones: con excepción de algunos antimicrobianos prescritos, la calidad de la prescripción en su mayoría resultó adecuada, la efectividad del tratamiento en su mayor parte fue buena y en general el uso racional de los antimicrobianos clasificó como alta y aceptable(AU)

Introduction: the misuse of drugs is a major health and economic problem, which is more sensitive in the case of children, the elderly and pregnant women. There is a great variety of medications and many of them do not have a definitive view of their safety, so the available information for prescription of drugs in pregnant women is poor. Objective: to assess quality in the rational use of antimicrobials in pregnant women admitted to Doña Leonor Perez Cabrera home for pregnant women during the 2007 to 2009 period. Methods: retrospective and descriptive, indication-prescription type study of drug use. The review of medical records allowed the study of social variables such as maternal age, gestational age on admission, occupation, diagnosis requiring the use of antimicrobials, most commonly used form of diagnosis and the most prescribed antimicrobials. Results: through the analysis of the variables and of treatment regimens, it was possible to evaluate the quality of prescribing antimicrobials. The results led to evidence that pregnant women aged 21 to 30, with gestational ages on admission between 21 and 30 weeks and working women predominated. The pathology that primarily motivated the use of antimicrobials was clinically diagnosed vaginal sepsis in almost all the cases and the most prescribed antimicrobials were azithromycin, metronidazole, clotrimazole and nystatin. Conclusions: the prescribing quality was generally adequate, the effectiveness of treatment was good and the rational use of antimicrobials was rated as high and acceptable(AU)

Utilización de antimicrobianos en el Hogar materno Doña Leonor Pérez durante el período 2007-2009 / Use of antimicrobials in Doña Leonor Pérez home for pregnant women in the 2007-2009 period

Introducción: el uso inadecuado de los medicamentos supone un importante problema sanitario y económico, más sensible en el caso de los niños, ancianos y las embarazadas. Existen una gran diversidad de medicamentos y sobre muchos de ellos no se tiene un criterio definitivo de su inocuidad, por lo que la información disponible para la prescripción de fármacos en mujeres embarazadas, es muy escasa. Objetivo: evaluar la calidad en el uso racional de antimicrobianos en las gestantes ingresadas en el Hogar materno Doña Leonor Pérez Cabrera durante el período 2007- 2009. Métodos: este trabajo constituye un estudio de utilización de medicamento del tipo indicación prescripción, retrospectivo y descriptivo. Se revisaron las historias clínicas y se obtuvieron las variables biosociales edad materna, edad gestacional al ingreso, ocupación, diagnóstico que precisó el uso de antimicrobianos, forma de diagnóstico más utilizado así como los antimicrobianos más prescritos. Resultados: a través del análisis de las variables y de los esquemas terapéuticos se pudo evaluar la calidad de la prescripción de los antimicrobianos. Los resultados obtenidos permitieron constatar que predominaron las gestantes con edades entre 21 y 30 años, con edades gestacionales al ingreso entre 21 y 30 semanas y trabajadoras. La patología que mayormente motivó la utilización de antimicrobianos fue la sepsis vaginal, diagnosticada de forma clínica en casi la totalidad de los casos y los antimicrobianos más prescritos fueron la azitromicina, metronidazol, clotrimazol y nistatina. Conclusiones: con excepción de algunos antimicrobianos prescritos, la calidad de la prescripción en su mayoría resultó adecuada, la efectividad del tratamiento en su mayor parte fue buena y en general el uso racional de los antimicrobianos clasificó como alta y aceptable

Introduction: the misuse of drugs is a major health and economic problem, which is more sensitive in the case of children, the elderly and pregnant women. There is a great variety of medications and many of them do not have a definitive view of their safety, so the available information for prescription of drugs in pregnant women is poor. Objective: to assess quality in the rational use of antimicrobials in pregnant women admitted to Doña Leonor Perez Cabrera home for pregnant women during the 2007 to 2009 period. Methods: retrospective and descriptive, indication-prescription type study of drug use. The review of medical records allowed the study of social variables such as maternal age, gestational age on admission, occupation, diagnosis requiring the use of antimicrobials, most commonly used form of diagnosis and the most prescribed antimicrobials. Results: through the analysis of the variables and of treatment regimens, it was possible to evaluate the quality of prescribing antimicrobials. The results led to evidence that pregnant women aged 21 to 30, with gestational ages on admission between 21 and 30 weeks and working women predominated. The pathology that primarily motivated the use of antimicrobials was clinically diagnosed vaginal sepsis in almost all the cases and the most prescribed antimicrobials were azithromycin, metronidazole, clotrimazole and nystatin. Conclusions: the prescribing quality was generally adequate, the effectiveness of treatment was good and the rational use of antimicrobials was rated as high and acceptable

Diseño y desarrollo de dos mascarillas faciales para el acné con quitina como sustancia bioactiva / Design and development of two face packs for the acne where chitin is the bioactive substance

La industria cosmética brinda importancia al logro de un cosmético más completo, un cosmecéutico, que represente seguridad en el mantenimiento de la salud y la belleza. En este trabajo se diseñaron y desarrollaron 2 mascarillas faciales para el acné con quitina al 2 por ciento como sustancia activa. Este polímero de origen marino posee propiedades hidratantes, filmógenas, ha sido demostrada su acción cicatrizante y antimicrobiana. Se realizaron 2 diseños de experimentos con mezcla D-optimal, y se estudió la incidencia de 3 componentes en ambas mascarillas: polyquaternium-7 (salcare SC91®) en una de las variantes a ensayar y carboximetil celulosa sódica (CMC) en la otra, propilenglicol y agua en ambos casos. Se analizaron en el tiempo el pH y la extensibilidad; resultó óptima la variante 4 del diseño (salcare SC91 2 por ciento , propilenglicol 2 por ciento y agua al 91,6 por ciento ) en el caso de la formulación con salcare y la variante 5 (CMC 2 por ciento , propilenglicol 4 por ciento y agua 89,6 por ciento ) en la formulación con CMC

The cosmetics industry attaches importance to the achievement of a more complete cosmetic, a cosmoceutic representing safety in health and beauty maintenance. In present paper were designed and developed two face packs for acne using 2 chitin as active substance. This polymer of marine origin has phylmogen and hydration properties and a demonstrated healing and antimicrobial action. Two experimental designs were carried out with a D-optimal mixture and also the study of incidence of three components of both face packs, polyquaaterium-7 (salcare SC91®) in one of the variants be assayed and sodium cellulose carboxymethyl (SCC) in the other one, 2 percent propylene glycol and 91,6 percent water) in the case of salcare formula and the 5 variant (2 percent SCC, 4 percent propylene glycol and 89,6 percent water) in SCC formula

Diseño y desarrollo de dos mascarillas faciales para el acné con quitina como sustancia bioactiva / Design and development of two face packs for the acne where chitin is the bioactive substance

La industria cosmética brinda importancia al logro de un cosmético más completo, un cosmecéutico, que represente seguridad en el mantenimiento de la salud y la belleza. En este trabajo se diseñaron y desarrollaron 2 mascarillas faciales para el acné con quitina al 2 por ciento como sustancia activa. Este polímero de origen marino posee propiedades hidratantes, filmógenas, ha sido demostrada su acción cicatrizante y antimicrobiana. Se realizaron 2 diseños de experimentos con mezcla D-optimal, y se estudió la incidencia de 3 componentes en ambas mascarillas: polyquaternium-7 (salcare SC91®) en una de las variantes a ensayar y carboximetil celulosa sódica (CMC) en la otra, propilenglicol y agua en ambos casos. Se analizaron en el tiempo el pH y la extensibilidad; resultó óptima la variante 4 del diseño (salcare SC91 2 por ciento , propilenglicol 2 por ciento y agua al 91,6 por ciento ) en el caso de la formulación con salcare y la variante 5 (CMC 2 por ciento , propilenglicol 4 por ciento y agua 89,6 por ciento ) en la formulación con CMC(AU)

The cosmetics industry attaches importance to the achievement of a more complete cosmetic, a cosmoceutic representing safety in health and beauty maintenance. In present paper were designed and developed two face packs for acne using 2 chitin as active substance. This polymer of marine origin has phylmogen and hydration properties and a demonstrated healing and antimicrobial action. Two experimental designs were carried out with a D-optimal mixture and also the study of incidence of three components of both face packs, polyquaaterium-7 (salcare SC91®) in one of the variants be assayed and sodium cellulose carboxymethyl (SCC) in the other one, 2 percent propylene glycol and 91,6 percent water) in the case of salcare formula and the 5 variant (2 percent SCC, 4 percent propylene glycol and 89,6 percent water) in SCC formula(AU)

Museo de la Farmacia Habanera y el adulto mayor: cita con el patrimonio farmacéutico y con temas de educación para la salud / Museum of Havanan Pharmacy and of third age: a an appointment with pharmaceutical heritage and with susbjects on health education

El Museo Farmacia Habanera es una institución que reúne y expone valiosos acervos históricos de la profesión farmacéutica de los siglos XIX y XX. Pertenece a la Dirección de Patrimonio Cultural de la Oficina del Historiador de la Ciudad de La Habana y está incorporado al Programa de Atención a la Tercera edad que dirige esta. Entre sus espacios habituales está la cita semanal con los adultos mayores procedentes de diversos círculos de abuelos de todos los municipios de la capital. El programa sociocultural organizado en estos encuentros habituales, comienza con un desayuno social, lo cual permite la identificación e intercambio de experiencias intergrupal. A continuación se les brinda una visita integral a la casa-museo dirigida por un especialista del museo y culmina el programa con charlas interactivas culturales, históricas y técnicas vinculadas a la farmacia. Durante el año 2009 con una frecuencia mensual, se incluyó en el programa de las charlas interactivas el tema de educación para la salud y la importancia de la adhesión al tratamiento prescrito para lograr una buena calidad de vida en la tercera edad. En los 10 encuentros realizados participaron 187 adultos mayores, procedentes de 7 municipios de la capital habanera. Estos se encontraban en buenas condiciones psicosociales, quienes demostraron independencia, capacidad de comunicación y expresión en la actividad coloquial interactiva que se establecía terminada la charla. Entre los participantes predominó el sexo femenino y el 46 por ciento se encontraba en el grupo de edades de 60 a 69 años. Las enfermedades crónicas que prevalecieron fueron la hipertensión arterial y la diabetis mellitus. Se detectó un bajo nivel de automedicación. Los encuentros finalizaban con la intervención de los participantes, quienes narraban sus experiencias sociales, recreativas y participativas que realizan dentro de los programas que se ejecutan en sus respectivos círculos de abuelos municipales

The Havana Pharmacy Musseum is an institution bringing togheter and exposes valuable historical heritages of pharmaceutical profession of XIX and XX centuries. It is sponsored by the Cultural Heritages Management from the Office of the Havana City Historian and it is annexed as part of the Third Age Care Program. Among the habitual spaces is the weekle meeting with elderlies from diverse grandparents circles from all capital municipalities of the. The sociocultural program organized in these habitual meetings starts qith a social breakfast allowing the intergroup indentification and exchange. Then, there is an integral visit to house-museum directed by a specialist of the institution and the program conclude with cultural, historical, interactive talks related to Pharmacy. During 2009 and with a montly frequency in the interctive program was included the subject on health education and the significance of the treatment adherence prescribed to achieve a good quality of life in third age. In the 10 meetings participate 187 elderlies from 7 Havana capital municipalities. Participants were in good psychosocial conditions demonstratiang independence, communitation and expression abilities in the debate at the end of the talk. Among participants there was predominance of female sex and the 46 percent was in 60-69 age groups. Chronic diseases prevailing were the high blood pressure and the diabetes mellitus. There was a low level of self-medication. Meetings finished with the participaton of all participants who recounted their social ,recreational and participatory experiences in the programs carried out in their respective municipal grandparents circles

Museo de la Farmacia Habanera y el adulto mayor: cita con el patrimonio farmacéutico y con temas de educación para la salud / Museum of Havanan Pharmacy and of third age: a an appointment with pharmaceutical heritage and with susbjects on health education

El Museo Farmacia Habanera es una institución que reúne y expone valiosos acervos históricos de la profesión farmacéutica de los siglos XIX y XX. Pertenece a la Dirección de Patrimonio Cultural de la Oficina del Historiador de la Ciudad de La Habana y está incorporado al Programa de Atención a la Tercera edad que dirige esta. Entre sus espacios habituales está la cita semanal con los adultos mayores procedentes de diversos círculos de abuelos de todos los municipios de la capital. El programa sociocultural organizado en estos encuentros habituales, comienza con un desayuno social, lo cual permite la identificación e intercambio de experiencias intergrupal. A continuación se les brinda una visita integral a la casa-museo dirigida por un especialista del museo y culmina el programa con charlas interactivas culturales, históricas y técnicas vinculadas a la farmacia. Durante el año 2009 con una frecuencia mensual, se incluyó en el programa de las charlas interactivas el tema de educación para la salud y la importancia de la adhesión al tratamiento prescrito para lograr una buena calidad de vida en la tercera edad. En los 10 encuentros realizados participaron 187 adultos mayores, procedentes de 7 municipios de la capital habanera. Estos se encontraban en buenas condiciones psicosociales, quienes demostraron independencia, capacidad de comunicación y expresión en la actividad coloquial interactiva que se establecía terminada la charla. Entre los participantes predominó el sexo femenino y el 46 por ciento se encontraba en el grupo de edades de 60 a 69 años. Las enfermedades crónicas que prevalecieron fueron la hipertensión arterial y la diabetis mellitus. Se detectó un bajo nivel de automedicación. Los encuentros finalizaban con la intervención de los participantes, quienes narraban sus experiencias sociales, recreativas y participativas que realizan dentro de los programas que se ejecutan en sus respectivos círculos de abuelos municipales(AU)

The Havana Pharmacy Musseum is an institution bringing togheter and exposes valuable historical heritages of pharmaceutical profession of XIX and XX centuries. It is sponsored by the Cultural Heritages Management from the Office of the Havana City Historian and it is annexed as part of the Third Age Care Program. Among the habitual spaces is the weekle meeting with elderlies from diverse grandparents circles from all capital municipalities of the. The sociocultural program organized in these habitual meetings starts qith a social breakfast allowing the intergroup indentification and exchange. Then, there is an integral visit to house-museum directed by a specialist of the institution and the program conclude with cultural, historical, interactive talks related to Pharmacy. During 2009 and with a montly frequency in the interctive program was included the subject on health education and the significance of the treatment adherence prescribed to achieve a good quality of life in third age. In the 10 meetings participate 187 elderlies from 7 Havana capital municipalities. Participants were in good psychosocial conditions demonstratiang independence, communitation and expression abilities in the debate at the end of the talk. Among participants there was predominance of female sex and the 46 percent was in 60-69 age groups. Chronic diseases prevailing were the high blood pressure and the diabetes mellitus. There was a low level of self-medication. Meetings finished with the participaton of all participants who recounted their social ,recreational and participatory experiences in the programs carried out in their respective municipal grandparents circles(AU)