Protein Thermal Stabilization and Delivery via Adsorption onto Porous Silicas

The cold chain is a problem that affects the whole pharmaceutical industry: in 2019, 45% of new FDA approved drugs required refrigeration or freezing. It is estimated that biopharma loses around 35 billion dollars every year due to failures in temperature control logistics. Proteins, the main components of vaccines, enzymes and many pharmaceuticals, are sensitive to denaturing and losing their conformational structure when exposed to thermal stress. This is why most pharmaceutical products require a temperature-controlled supply chain to ensure that heat exposure does not disrupt the intermolecular interaction needed to maintain protein structure and activity. The aim of this study is to develop a more sustainable technology via protein adsorption onto mesoporous silica, in order to improve the protein’s thermal stability. Two proteins were investigated: Invasion Plasmid Antigen D (IpaD) and Lactoferrin (Lf). Circular dichroism (CD) confirmed that the adsorbed IpaD and Lf after the heat treatment maintained a native secondary structure rich in α-helical content. In contrast, the unprotected proteins after heat treatment lost their secondary structure. Isotherms modeled using Langmuir, Freundlich and Temkin models demonstrated that the adsorption of IpaD and Lf onto silicas is best fit by the Langmuir model. Silica pore size was determined to be a key factor in protein adsorption. If pores are less than 15 nm, adsorption is negligible. If the pores are between 15-25 nm, then monolayer coverage is achieved and IpaD is protected from thermal denaturing. If pores are larger than 25 nm the adsorption is multilayer coverage and it is easier to remove the protein from the silica due to a less developed hydrogen bond network. In addition, this work investigates the protein release from silica using non-ionic detergents (Triton X-100, Tween 20, 40 and 80). Lactoferrin desorption tests with non-ionic detergents showed a trend for increased Lf recovery with increased detergent hydrophobicity. Triton X-100 and Tween 80 were the best detergents for Lf desorption, recovering over 80% of the initial protein. This research provides strong evidence that proteins can be thermally stabilized on mesoporous silica and efficiently released from the silica using non-ionic detergents

Attitudes, health lifestyle behaviors and cardiometabolic risk factors among relatives of individuals with type 2 diabetes mellitus (actitudes, comportamientos de estilo de vida y factores de riesgo cardio-metabólicos en familiares de individuos con diabetes melitus tipo 2)

Objetivo: Describir y comparar actitudes, conductas en el estilo de vida y factores de riesgo cardio-metabólicos en individuos que convivan o no con familiares diagnosticados con diabetes mellitus tipo 2 (DM2). Metodología: Análisis secundario de base de datos procedente de un estudio de implementación llevado a cabo en Perú. Las variables de interés fueron actitudes de cambio de estilo de vida (intención de pérdida de peso, aumento de actividad física, reducción en el consumo de sal, etc), conductas en el estilo de vida (tabaquismo, consumo de bebidas alcohólicas y actividad física) y factores de riesgo cardio-metabólicos (sobrepeso [índice de masa corporal >25 kg/m] e hipertensión); mientras que la variable de exposición fue la presencia de al menos un familiar diagnosticado de DM2 dentro del hogar. Se utilizó un modelo de Regresión logística multinivel de efectos mixtos para estimar la razón de probabilidades (OR) y un intervalo de confianza de 95% (IC 95%). Resultados: Se obtuvo un total de 2298 registros, 1134 (49.4%) varones, con una media de edad de 43.3 años (DE 0.36), todos sometidos a análisis. No se encontró evidencia de diferencia en actitudes de cambio de estilo de vida, tabaquismo, consumo de bebidas alcohólicas, niveles de actividad física e hipertensión entre los individuos con o sin familiar con DM2. El modelo multivariado señaló que el sobrepeso era 63% más frecuente entre individuos con familiares diabéticos (OR = 1.63; 95% IC: 1.3-2.61). Conclusiones: Individuos con familiares con DM2 tiene una probabilidad más alta de tener sobrepeso en comparación a personas que no cuentan con familiares diabéticos dentro del hogar. La ausencia de diferencias en actitudes de cambio y conductas en el estilo de vida resaltan la necesidad de incluir a los familiares de pacientes diabéticos dentro de las estrategias de prevención y manejo de DM2.Objective: To describe and compare attitudes, lifestyle behaviors, and cardiometabolic risk factors between individuals with and without a relative with type 2 diabetes mellitus (T2DM) living in the same household. Methods: A secondary analysis of baseline data from an implementation study in Peru was conducted. The outcomes were attitudes towards changing lifestyle behaviors (e.g. intentions towards losing weight, increasing physical activity, reducing salt consumption, etc), profiles of health lifestyle behaviors (e.g. daily smoking, heavy drinking, and physical activity), and cardiometabolic risk factors (e.g., overweight [body mass index ≥25 kg/m2] and hypertension); whereas the exposure was the presence of at least one relative with known diagnosis of T2DM living in the same household. Multilevel logistic mixed effect regression models were used to estimate odds ratios (OR) and 95% confidence intervals (95% CI). Results: A total of 2298 records, 1134 (49.4%) males, mean age 43.3 (SD: 17.2) years, were analyzed. There was no evidence of a difference in lifestyle-changing attitudes, smoking, alcohol drinking, physical activity levels, and hypertension between individuals with and without relatives with T2DM. Overweight was 63% more common among individuals having a relative with a T2DM in multivariable model (OR = 1.63; 95% CI: 1.03–2.61). Conclusions: Individuals with relatives with T2DM have higher probabilities of being overweight compared to those who did not have relatives with T2DM in the same household. The absence of differences on lifestyle-related attitudes and behaviors highlight the need of involving relatives of patients with T2DM on intervention strategies to further enhance diabetes prevention and management efforts.Tesi

Maternal Neural Responses to Infant Cries and Faces: Relationships with Substance Use

Substance abuse in pregnant and recently post-partum women is a major public health concern because of effects on the infant and on the ability of the adult to care for the infant. In addition to the negative health effects of teratogenic substances on fetal development, substance use can contribute to difficulties associated with the social and behavioral aspects of parenting. Neural circuits associated with parenting behavior overlap with circuits involved in addiction (e.g., frontal, striatal, and limbic systems) and thus may be co-opted for the craving/reward cycle associated with substance use and abuse and be less available for parenting. The current study investigates the degree to which neural circuits associated with parenting are disrupted in mothers who are substance-using. Specifically, we used functional magnetic resonance imaging to examine the neural response to emotional infant cues (faces and cries) in substance-using compared to non-using mothers. In response to both faces (of varying emotional valence) and cries (of varying distress levels), substance-using mothers evidenced reduced neural activation in regions that have been previously implicated in reward and motivation as well as regions involved in cognitive control. Specifically, in response to faces, substance users showed reduced activation in prefrontal regions, including the dorsolateral and ventromedial prefrontal cortices, as well as visual processing (occipital lobes) and limbic regions (parahippocampus and amygdala). Similarly, in response to infant cries, substance-using mothers showed reduced activation relative to non-using mothers in prefrontal regions, auditory sensory processing regions, insula and limbic regions (parahippocampus and amygdala). These findings suggest that infant stimuli may be less salient for substance-using mothers, and such reduced saliency may impair developing infant-caregiver attachment and the ability of mothers to respond appropriately to their infants

From “trust” to “trustworthiness”: Retheorizing dynamics of trust, distrust, and water security in North America

Assumptions of trust in water systems are widespread in higher-income countries, often linked to expectations of “modern water.” The current literature on water and trust also tends to reinforce a technoscientific approach, emphasizing the importance of aligning water user perceptions with expert assessments. Although such approaches can be useful to document instances of distrust, they often fail to explain why patterns differ over time, and across contexts and populations. Addressing these shortcomings, we offer a relational approach focused on the trustworthiness of hydro-social systems to contextualize water-trust dynamics in relation to broader practices and contexts. In doing so, we investigate three high-profile water crises in North America where examples of distrust are prevalent: Flint, Michigan; Kashechewan First Nation; and the Navajo Nation. Through our theoretical and empirical examination, we offer insights on these dynamics and find that distrust may at times be a warranted and understandable response to experiences of water insecurity and injustice. We examine the interconnected experiences of marginality and inequity, ontological and epistemological injustice, unequal governance and politics, and histories of water insecurity and harm as potential contributors to untrustworthiness in hydro-social systems. We close with recommendations for future directions to better understand water-trust dynamics and address water insecurity

Development of Silica-Immobilized Vaccines for Improving Thermo-Tolerance and Shelf-Life

Introduction. It is estimated that 50% of vaccines produced annu- ally are wasted because effectivity is dependent on protein structure and heat exposure disrupts the intermolecular interactions that maintain this structure. Since 90% of vaccines require a temperature- controlled supply chain, it is necessary to create a cold chain system to minimize vaccine waste. We have developed a more sustainable technology via the adsorption of Invasion Plasmid Antigen D (IpaD) onto mesoporous silica gels, improving the thermal stability of pro- tein-based therapeutics. Methods.xThe solution depletion method using UV-Vis was uti- lized to study the adsorption of IpaD onto silica gels. The silica-IpaD complex is heated above the denaturing temperature of the protein and then the IpaD is removed using N,N-Dimethyldodecylamine N-oxide (LDAO) and their secondary structure is tested using cir- cular dichroism (CD). Results. Pore diameter, pore volume and surface area were charac- terized for seven different silica gels. Silica gels designated as 6389, 6378, and 6375 had an adsorption percentage above 95% at pore volumes of 2.2, 2.8 and 3.8 cm3 mg-1, respectively. CD analyses con- firmed that the adsorbed IpaD after the heat treatment displayed a similar “W” shape CD signal as the native IpaD, indicating the con- servation of α-helices. In contrast, the unprotected IpaD after being exposed to high temperature shows a flat CD signal, demonstrating the loss of secondary structure. Conclusion. We have successfully increased the thermo-tolerance for IpaD using mesoporous silica and continue to further optimize mesoporous silica’s physiochemical properties to improve adsorption and desorption yields

WAYLLA KAY – Galletas saludables a base de cushuro, y frutos nativos del Perú

Actualmente, muchas personas toman en cuenta la importancia del cuidado de la salud, lo cual ha generado una mayor demanda de productos que brindan beneficios para su cuidado y la prevención de las enfermedades. De tal manera, cada vez son más las personas que buscan consumir productos que contengan vitaminas, minerales y proteínas, para que así puedan reforzar su sistema inmunológico, con el fin de prevenir algunas enfermedades vinculadas con las bajas defensas, la falta de hemoglobina en sangre o deficiencia de vitaminas. Por tal motivo, se asume que una buena alimentación es fundamental para mantener un cuerpo sano y una vida equilibrada tanto física como mentalmente. No obstante, algunas veces resulta complicado seguir un régimen saludable y provoca salir de la rutina al darse un gusto con algunos productos altos en azúcares o altos en sodio. Por ello, el problema de esta situación es cuando no se sabe cómo balancear el consumo de todos los alimentos, y se ingieren altas cantidades de estos insumos dañinos, estos a largo plazo pueden generar enfermedades como la diabetes, el colesterol, la hipertensión. A raíz de ello, el presente proyecto tiene como finalidad estudiar la viabilidad de nuestro modelo de negocio llamado Waylla galletas. Para la ejecución de este proyecto, cada integrante propuso diversas ideas de negocio innovadoras, predominando entre ellas la venta las galletas de cushuro, un alga andina que aporta vitaminas B5 y B8, que se considera uno de los alimentos más nutritivos de los andes peruanos y que refuerza el sistema inmunológico. Complemento de ello, se consideró para la preparación de las galletas, agregarle frutos nativos de las 3 regiones del Perú, como son el camu camu, el arándano, la fresa y la lúcuma. Agregando a lo anterior, el camu camu es un fruto que proviene de la Selva Peruana y contiene un porcentaje mayor de vitamina C que cualquier otro alimento, además proporciona aminoácidos esenciales y actúa como una fuente de minerales (hierro y calcio). Seguidamente, los arándanos son sembrados en las regiones de Loreto y Ucayali pertenecientes a la Selva del Perú. Estos frutos son conocidos por poseer antocianina, la cual ayuda a revitalizar la memoria, es un antioxidante natural y reduce los niveles de azúcar en la sangre. De igual manera, la fresa es cultivada en distintas zonas del “norte chico” de lima e incluso en Arequipa y Cusco que pertenecen a la Sierra del Perú. Esta fruta presenta un alto contenido de antioxidantes, minerales como magnesio, manganeso y potasio, cuenta con vitamina C, B2 y B3 y es baja en azúcar. Igualmente, esta cantidad de beneficios previenen las enfermedades cardiovasculares, favorecen la capacidad antiviral y retrasan el envejecimiento cerebral. De modo similar, la lúcuma es cultivada en Huaral y Huarochirí, proveniente de la Sierra costera del Perú, esta fruta tiene propiedades antioxidantes y antiinflamatorias, presentando un gran potencial para prevenir enfermedades como cáncer y Alzheimer. Finalmente, este producto está dirigido a personas entre 18 a 55 años que vivan en Lima Moderna, que se preocupen por cuidar su salud y prevenir enfermedades. Por consiguiente, nuestra meta es ingresar al mercado concientizando a la población sobre la importancia de consumir productos saludables y, sobre todo, posicionarse como la mejor empresa dentro del rubro.These days, many people value the importance of their health, this has brought about a greater demand for products that cultivate good hearth and prevent disease. There are more and more people looking to consume products that have vitamins, minerals and proteins, so that they can then re-enforce their immune system, with the ends of preventing illnesses linked to low defenses, low blood hemoglobin, or vitamins deficiency. For this reason, it is assumed that a good diet is essential to maintain a healthy body and a balanced life both physically and mentally. Nevertheless, it is sometimes difficult to follow a healthy regimen. This can result in deviating from the routine and end up eating some products high in sugar or high in sodium. When one doesn’t know how to balance all the ingredients, and you can eat high amounts of these harmful ingredients. Long term, they can result in diseases such as diabetes, high cholesterol, and hypertension. As a result, the current project has finalized a viability of our model the business called Waylla cookies. For the execution of this project, each member proposed various innovative business ideas. Most predominate among the ideas is the sale of cushuro cookies. Cushuro is an Andean alga that contains B5 and B8 and is considered one of the most nutritious ingredients of the Peruvian andes that reinforces the immune system. Complementing this, it was considered for the preparation of the cookies, to add native fruits of the 3 regions of Peru, such as camu camu, blueberry, strawberry and lucuma. Adding to the above, camu camu is a fruit that comes from the Peruvian jungle and contains a higher percentage of vitamin C than any other food, it also provides essential amino acids and is also a good source of minerals (iron and calcium). Next, the blueberries are planted in the regions of Loreto and Ucayali belonging to the Peruvian jungle. These fruits are known for their antioxidants, which helps revitalize memory, it is a natural antioxidant and reduces blood sugar levels. In the same way the strawberry, is cultivated in different areas including "norte chico" of Lima, Arequipa, and Cuzco, which are in the Sierra of Peru. This fruit contains a high amounts of antioxidants, minerals like magnesium, manganese, and potassium, it counts with vitamins C, B2, B3 and low in sugar. Likewise, this amount of benefits prevents cardiovascular diseases, cultivates antiviral capacity, and delays brain aging. Similarly, the lucuma is cultivated in Huaral and Huarochiri, coming from the costal Siera of Peru, this fruit has antioxidant and anti-inflammatory properties, presenting a great potential for preventing illnesses like cancer and Alzheimer’s. Finally, this product is aimed at people between 18 to 55 years old who live in modern Lima, who care about taking care of their health and preventing diseases. Consequently, our goal is to enter the market by making the population aware of the importance of consuming healthy products and, above all, to position ourselves as the best company in the field.Trabajo de investigació

Differences in small intestinal apparent amino acid digestibility of raw bovine, caprine, and ovine milk are explained by gastric amino acid retention in piglets as an infant model

BackgroundThe rate of stomach emptying of milk from different ruminant species differs, suggesting that the small intestinal digestibility of nutrients could also differ across these milk types.ObjectiveTo determine the small intestinal amino acid (AA) digestibility of raw bovine, caprine, and ovine milk in the piglet as an animal model for the infant.MethodsSeven-day-old piglets (n = 12) consumed either bovine, caprine, or ovine milk diets for 15 days (n = 4 piglets/milk). On day 15, fasted piglets received a single meal of fresh raw milk normalized for protein content and containing the indigestible marker titanium dioxide. Entire gastrointestinal tract contents were collected at 210 min postprandially. Apparent AA digestibility (disappearance) in different regions of the small intestine was determined.ResultsOn average, 35% of the dietary AAs were apparently taken up in the small intestine during the first 210 min post-feeding, with 67% of the AA digestibility occurring in the first quarter (p ≤ 0.05) and 33% in the subsequent two quarters. Overall, except for isoleucine, valine, phenylalanine, and tyrosine, the small intestinal apparent digestibility of all AAs at 210 min postprandially in piglets fed ovine milk was, on average, 29% higher (p ≤ 0.05) than for those fed bovine milk. Except for lysine, there was no difference in the apparent digestibility (p > 0.05) of any AAs between piglets fed caprine milk or ovine milk. The apparent digestibility of alanine was higher (p ≤ 0.05) in piglets fed caprine milk than those fed bovine milk. When apparent digestibility was corrected for gastric AA retention, only small differences in the small intestinal apparent digestibility of AAs were observed across milk types.ConclusionBovine, caprine and ovine milk had different apparent small intestinal AA digestibility at 210 min postprandially. When corrected for gastric AA retention, the differences in apparent digestibility across species largely disappeared. The apparent AA digestibility differed across small intestinal locations

Metabolite profiling of peripheral blood plasma in pigs in early postnatal life fed whole bovine, caprine or ovine milk

Ruminants’ milk is commonly used for supplying nutrients to infants when breast milk is unavailable or limited. Previous studies have highlighted the differences between ruminants’ milk composition, digestion, absorption, and fermentation. However, whether consuming different ruminants’ milk impact the appearance of the circulatory blood metabolites in the early postnatal life is not well understood. The analysis conducted here aimed to determine the effect of feeding exclusively whole milk from bovine, caprine or ovine species to pigs, approximately 7 days-old for 15 days, on circulatory blood plasma metabolites. Relative intensities of plasma metabolites were detected using a liquid chromatography-mass spectrometry based metabolomic approach. Seven polar and 83 non-polar (lipids) metabolites in plasma were significantly different (false discovery rate < 0.05) between milk treatments. These included polar metabolites involved in amino acid metabolism and lipids belonging to phosphatidylcholine, lysophosphatidylcholine, sphingomyelin, and triglycerides. Compared to the caprine or bovine milk group, the relative intensities of polar metabolites and unsaturated triglycerides were higher in the peripheral circulation of the ovine milk group. In contrast, relative intensities of saturated triglycerides and phosphatidylcholine were higher in the bovine milk group compared to the ovine or caprine milk group. In addition, correlations were identified between amino acid and lipid intake and their appearance in peripheral blood circulation. The results highlighted that consuming different ruminants’ milk influences the plasma appearance of metabolites, especially lipids, that may contribute to early postnatal life development in pigs

Warming, drought, and disturbances lead to shifts in functional composition: A millennial-scale analysis for Amazonian and Andean sites

Tropical forests are changing in composition and productivity, probably in response to changes in climate and disturbances. The responses to these multiple environmental drivers, and the mechanisms underlying the changes, remain largely unknown. Here, we use a functional trait approach on timescales of 10,000¿years to assess how climate and disturbances influence the community-mean adult height, leaf area, seed mass, and wood density for eight lowland and highland forest landscapes. To do so, we combine data of eight fossil pollen records with functional traits and proxies for climate (temperature, precipitation, and El Niño frequency) and disturbances (fire and general disturbances). We found that temperature and disturbances were the most important drivers of changes in functional composition. Increased water availability (high precipitation and low El Niño frequency) generally led to more acquisitive trait composition (large leaves and soft wood). In lowland forests, warmer climates decreased community-mean height probably because of increased water stress, whereas in highland forests warmer climates increased height probably because of upslope migration of taller species. Disturbance increased the abundance of acquisitive, disturbance-adapted taxa with small seeds for quick colonization of disturbed sites, large leaves for light capture, and soft wood to attain fast height growth. Fire had weak effects on lowland forests but led to more stress-adapted taxa that are tall with fast life cycles and small seeds that can quickly colonize burned sites. Site-specific analyses were largely in line with cross-site analyses, except for varying site-level effects of El Niño frequency and fire activity, possibly because regional patterns in El Niño are not a good predictor of local changes, and charcoal abundances do not reflect fire intensity or severity. With future global changes, tropical Amazonian and Andean forests may transition toward shorter, drought- and disturbance-adapted forests in the lowlands but taller forests in the highlands.We thank various funding sources. M.T.v.d.S. was supported by the Rubicon research program with project number 019.171LW.023 and the Veni research program with project number NWO-VI.Veni.192.027, both funded by the Netherlands Organisation for Scientific Research (NWO); M.T.v.d.S. and L.P. by the European Research Council Advanced Grant PANTROP 834775; M.B.B. by the National Science Foundation 621 (grant # EAR1338694, BCS0926973 and 1624207), the Belmont Forum, the National Aeronautics 622 and Space Administration (grant no. NNX14AD31G), and National Geographic Society (grant no. 8763-10); S.G.A.F. by Trond Mohn Stiftelse (TMS) and University of Bergen (Grant No. TMS2022STG03/Past, Present and Future of Alpine Biomes Worldwide); H.H. by the Netherlands Organization for Tropical Research (grant WB 84-636) to study Lake La Cocha; S.Y.M. and W.D.G. by the European Commission (Marie Curie Fellowship 792197); C.N.H.M. by the European Research Council Starting Grant StG 853394 (2019) and the NWO-ALWOP.322; E.M. by the Natural Environment Research Council (NERC) of UK (grant NE/J018562/1), and for radiocarbon dating by the NERC Radiocarbon Facility NRCF010001 (allocation number 1682.1112); M.d.N.N. by the National Science Foundation (NSF)—DEB 1260983, EAR 1338694 and 1624207. All work for Lake Kumpaka was conducted under Ecuadorian Collection Permit 08-620 2017-IC and for Lake Pindo under Permit 14-2012-IC-FLO-DPAP-MA