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1.
Int J Oral Maxillofac Implants ; 36(2): 295-306, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33909719

RESUMEN

PURPOSE: The aim of this study was to explore the influence of different bone grafts, demineralized freeze-dried bone allograft (DFDBA, OraGraft), and deproteinized bovine bone mineral (DBBM, Bio-Oss) implanted in mouse calvaria defects on gene expression. MATERIALS AND METHODS: Male C57BL/6MLac mice were separated into three groups as follows: group 1-defect without graft as control, group 2-DFDBA, and group 3-DBBM. Affymetrix DNA microarrays were used to characterize gene expression in bone after 3 months of graft healing. Differential expression of designated genes discovered by microarray analysis was confirmed using real-time polymerase chain reaction (PCR) and immunohistochemistry. RESULTS: Compared with normal bone healing, 355 and 1,108 coding genes of bone grafted with DFDBA were upregulated and downregulated, respectively. The upregulated genes were mainly involved in chemokine signaling, macrophage activity, osteoclast activity, cytokine expression, T-cell receptor signaling, apoptosis, and MAPK signaling. The downregulated genes were predominantly involved in calcium regulation in cardiac cells, chemokine signaling, MAPK signaling, and adipogenesis. A total of 306 and 817 coding genes of bone grafted with DBBM were upregulated and downregulated, respectively. The upregulated genes were mainly involved in osteoclast activity, chemokine signaling, B cell receptor signaling, macrophage activity, and signaling of T-cell receptor, MAPK, IL-5, and IL-1. The downregulated genes were predominantly involved in calcium regulation in the cardiac cell and osteoclast activity. Real-time PCR revealed that the DFDBA and DBBM groups showed a higher mRNA level of MMP12, Bcl2A1, S100A4, and Postn compared with the control (P < .05). Histology showed that, compared with the control, the volume of new bone was higher in both types of bone grafts. Immunohistochemistry using an MMP12 antibody confirmed the microarray results because the MMP12 immunoreactivity intensified, and a positive expression of MMP12 increased significantly in the DFDBA and DBBM groups. CONCLUSION: Both DFDBA and DBBM had a gene expression network involved in new bone formation, which coincided with an increased expression of MMP-12 and osteoclast activity. Both types of graft materials appeared to connect with genes that stimulate bone remodeling at 3 months of bone grafting.


Asunto(s)
Sustitutos de Huesos , Implantes Dentales , Aloinjertos , Animales , Trasplante Óseo , Bovinos , Liofilización , Expresión Génica , Xenoinjertos , Masculino , Ratones , Ratones Endogámicos C57BL
2.
Artículo en Chino | MEDLINE | ID: mdl-33781042

RESUMEN

Objective: To develop the certified reference material of cadmium in human urine (freeze-dried) . Methods: Urine were collected from normal human, and then the urine samples were prepared by adding standard, mixing, packing and freeze-drying. After homogeneity test and stability test, the urine samples were determined by two methods in seven independent laboratories, the uncertainty of the certified reference material was evaluated. Results: Take at least 0.20 ml solution of the certified reference material per use, and the homogeneity is good. The freeze-dried materials can be kept stable for 12 months at ≤4 °C, and the re-dissolved reference materials can be stabilized for 7 days at 4 °C. The standard values and uncertainty of the two levels were (5.34±0.34) µg/L and (15.47±0.72) µg/L, respectively. Conclusion: The standards met the requirements of national standards, and can be used to verify the methods of detecting cadmium in human urine and to detect the samples.


Asunto(s)
Cadmio , Laboratorios , Liofilización , Humanos , Estándares de Referencia
3.
Int J Nanomedicine ; 16: 2013-2044, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33727812

RESUMEN

Background: Sulpiride (SUL), is a selective antidopaminergic drug that had extensive biological activities. However, its sparingly aqueous solubility and limited gastrointestinal permeability lead to scanty oral bioavailability which hinders its clinical efficacy. Objective: SUL-loaded lipospheres (SUL-LPS) were designed to serve as an oral biocompatible nanovector for improving SUL permeability as well as conquering its low oral absorption and then in turn enhancing its antidepressant action. Methods: SUL-LPS were fabricated via two processing techniques namely, melt emulsification and solvent evaporation. The impact of different lipid cores, phospholipid shells together with various surfactant concentrations and types on the lipospheres properties were screened. Detailed physicochemical elucidations were performed followed by ex vivo permeation appraisal using the non-everted intestine model. The pharmacokinetic parameters of SUL-LPS, free SUL and marketed product were assessed following oral administration to healthy rats. Reserpine-induced depression rat model was used to assess the antidepressant action of SUL-LPS on which full behavioural and biochemical analysis was conducted. Safety attributes of nanoencapsulated SUL on the brain and other internal organs were evaluated. Results: The optimum LPS revealed an excellent nanosize with a narrow PdI, negative zeta potential and acceptable entrapment efficiency of 68.62 nm, 0.242, -30.4 mV and 84.12%, respectively. SUL-LPS showed a sustained release pattern and 2.1-fold enhancement in the intestinal permeation parameters with low mucin interaction. Oral pharmacokinetic appraisal exhibited that LPS provided 3.4-fold improvement in SUL oral bioavailability together with long-circulating properties, relative to the free drug. Pharmacodynamic study confirmed the superior antidepressant action of SUL-LPS as evident by 1.6 and 1.25-fold elevation in the serotonin and dopamine expressions, respectively. Meanwhile, nanotoxicological appraisal proved the biocompatibility of SUL-LPS upon repetitive oral administration. Conclusion: Rationally designed lipospheres hold promising in vitro and in vivo characteristics for efficient delivery of SUL with high oral bioavailability, antidepressant activity together with a good safety profile.


Asunto(s)
Antidepresivos/farmacología , Lípidos/química , Nanopartículas/química , Sulpirida/administración & dosificación , Sulpirida/farmacología , Administración Oral , Animales , Materiales Biocompatibles/química , Disponibilidad Biológica , Cromatografía Líquida de Alta Presión , Composición de Medicamentos , Liberación de Fármacos , Liofilización , Masculino , Mucinas/química , Nanopartículas/ultraestructura , Neurotransmisores/metabolismo , Especificidad de Órganos/efectos de los fármacos , Tamaño de la Partícula , Permeabilidad , Ratas Sprague-Dawley , Ratas Wistar , Sulpirida/química , Sulpirida/farmacocinética , Porcinos
4.
Methods Mol Biol ; 2278: 101-115, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33649951

RESUMEN

This chapter describes some of the available methods to assess EPS production in bifidobacteria, being largely based on those developed for the same purpose for members of the lactic acid bacteria group. The first step is detection of putative EPS-producing bifidobacteria based on a mucoid and/or ropy phenotype. Next, a basic procedure is described for the isolation of the glycan polymer based on the release from bifidobacterial cells grown and collected from the surface of agar-MRSc ("crude EPS"), followed by a purification procedure intended to remove other bacterial macromolecules (DNA and proteinaceous material) to generate "purified EPS." Finally, several methods used for quantification and physical-chemical characterization of isolated/purified polysaccharide are outlined.


Asunto(s)
Bifidobacterium/química , Polisacáridos Bacterianos/aislamiento & purificación , Técnicas de Cultivo de Célula/métodos , Centrifugación/métodos , Precipitación Química , Diálisis/métodos , Liofilización/métodos , Polisacáridos/aislamiento & purificación
5.
Molecules ; 26(4)2021 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-33669650

RESUMEN

Apples are an important source of biologically active compounds. Consequently, we decided to model hard gelatin capsules with lyophilized apple powder by using different excipients and to evaluate the release kinetics of phenolic compounds. The apple slices of "Ligol" cultivar were immediately frozen in a freezer (at -35°C) with air circulation and were lyophilized with a sublimator at the pressure of 0.01 mbar (condenser temperature, -85°C). Lyophilized apple powder was used as an active substance filled into hard gelatin capsules. We conducted capsule disintegration and dissolution tests to evaluate the quality of apple lyophilizate-containing capsules of different encapsulating content. Individual phenolic compounds can be arranged in the following descending order according to the amount released from the capsules of different compositions: chlorogenic acid > rutin > avicularin > hyperoside > phloridzin > quercitrin > (-)-epicatechin > isoquercitrin. Chlorogenic acid was the compound that was released in the highest amounts from capsules of different encapsulating content: its released amounts ranged from 68.4 to 640.3 µg/mL. According to the obtained data, when hypromellose content ranged from 29% to 41% of the capsule mass, the capsules disintegrated within less than 30 min, and such amounts of hypromellose did not prolong the release of phenolic compounds. Based on the results of the dissolution test, the capsules can be classified as fast-dissolving preparations, as more than 85% of the active substances were released within 30 min.


Asunto(s)
Productos Biológicos/análisis , Cápsulas/química , Liofilización , Malus/química , Etanol/química , Gelatina/química , Glicósidos/química , Fenoles/análisis , Fenoles/química , Polvos
6.
Carbohydr Polym ; 260: 117765, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712123

RESUMEN

Chitosan (CS) combined with hydroxyapatite (HA) was injected into a composite braid, and a hierarchical pore structure scaffold was obtained by freeze drying and cold atmospheric plasma (CAP) technology. The CS/HA/braid scaffold with hierarchical pore structure was analyzed and characterized by scanning electronic microscopy, Fourier transform infrared spectroscopy, true color confocal microscopy, improved liquid replacement method, and phosphate buffer solution immersion. The mechanical properties and degradation ability of the scaffold were evaluated through compression test and degradation test. Results showed that HA addition endowed the core of the scaffold with macroscopic pore sizes of 80-180 µm, and CAP treatment endowed the shell of the scaffold with microscopic pore sizes ≤10 µm. All scaffolds exhibited high porosity and swelling rates of ≥80 % and ≥300 %, respectively. The scaffold with a hierarchical pore structure had good mechanical properties and twice the degradation rate. In addition, the treated scaffold precipitated intact spherical HA crystals. Under the synergistic effect of HA and CAP treatment, scaffolds achieved 277.6 % cell viability compared with pure CS scaffold. Overall, this method was feasible for preparing bone scaffolds with hierarchical pore structure for potential bone tissue engineering.


Asunto(s)
Quitosano/química , Durapatita/química , Ingeniería de Tejidos , Andamios del Tejido/química , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fuerza Compresiva , Liofilización , Ratones , Porosidad
7.
Carbohydr Polym ; 260: 117767, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712125

RESUMEN

Wound healing is a dynamic and intricate process, and newly dressings are urgently needed to promote wound healing over the multiple stages. Herein, two water-soluble adenine-modified chitosan (CS-A) derivatives were synthesized in aqueous solutions and freeze-dried to obtain porous sponge-like dressings. The novel derivatives displayed antibacterial activities against S. aureus and E. coli. Moreover, CS-A derivatives demonstrated excellent hemocompatibility and cytocompatibility, as well as promoted the proliferation of the wound cells by shortening the G1 phase and improving DNA duplication efficiency. The ability of CS-A sponges to promote wound healing was studied in a full-thickness skin defect model. The histological analysis and immunohistochemical staining showed that the wounds treated with CS-A sponges displayed fewer inflammatory cells, and faster regeneration of epithelial tissue, collagen deposition and neovascularization. Therefore, CS-A derivatives have potential application in wound dressings and provide new ideas for the design of multifunctional biomaterials.


Asunto(s)
Adenina/química , Materiales Biocompatibles/química , Quitosano/química , Animales , Vendajes , Materiales Biocompatibles/farmacología , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Liofilización , Masculino , Ratones , Porosidad , Ratas , Ratas Sprague-Dawley , Piel/efectos de los fármacos , Piel/patología , Cicatrización de Heridas/efectos de los fármacos
8.
Carbohydr Polym ; 260: 117843, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712117

RESUMEN

In this study, trehalose (TRE) was added to prepare whey protein concentrate (WPC)/pullulan (PUL)/TRE hydrogel and the hydrogel was used as the wall material to improve the viability of encapsulated L. plantarum during freeze drying and storage. The optimum conditions were 5.0 % TRE concentration and 1:4 (v:v) of the ratio of L. plantarum suspension to the hydrogel. Under these conditions, the survival rates of L. plantarum were 94.36 ± 1.06 % after freeze drying and 97.02 ± 0.30 % after storage for 240 d at 4 °C. Interactions and rheological properties of WPC/PUL/TRE hydrogel were also studied. The results showed TRE reduced storage modulus (G') of the hydrogel and weakened hydrophobic interactions, disulfide and hydrogen bonds between proteins and polysaccharides, which was not conducive to hydrogel formation. In addition, the excellent water-holding capacity of WPC/PUL/TRE hydrogel was found by the drying kinetic experiment.


Asunto(s)
Glucanos/química , Hidrogeles/química , Lactobacillus plantarum/fisiología , Trehalosa/química , Proteína de Suero de Leche/química , Liofilización , Hidrogeles/farmacología , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Viabilidad Microbiana/efectos de los fármacos , Reología , Compuestos de Sulfhidrilo/química
9.
Artículo en Inglés | MEDLINE | ID: mdl-33671763

RESUMEN

BACKGROUND: Human milk banks have a pivotal role in provide optimal food for those infants who are not fully breastfeed, by allowing human milk from donors to be collected, processed and appropriately distributed. Donor human milk (DHM) is usually preserved by Holder pasteurization, considered to be the gold standard to ensure the microbiology safety and nutritional value of milk. However, as stated by the European Milk Banking Association (EMBA) there is a need to implement the improvement of the operating procedure of human milk banks including preserving and storing techniques. AIM: The purpose of this study was to assess the effectiveness and safety of the selected new combination of methods for preserving donor human milk in comparison with thermal treatment (Holder pasteurization). METHODS: We assessed (1) the concentration of bioactive components (insulin, adiponectin, leptin, activity of pancreatic lipase, and hepatocyte growth factor) and (2) microbiological safety in raw and pasteurized, high-pressure processed and lyophilization human breast milk. RESULTS: The combination of two techniques, high-pressure processing and freeze-drying, showed the best potential for preserving the nutritional value of human milk and were evaluated for microbiological safety. Microbiological safety assessment excluded the possibility of using freeze-drying alone for human milk sample preservation. However, it can be used as a method for long-term storage of milk samples, which have previously been preserved via other processes. CONCLUSION: The results show that high-pressure treatment is the best method for preservation that ensures microbiological safety and biological activity but subsequent freeze-drying allowed long-term storage without loss of properties.


Asunto(s)
Bancos de Leche , Animales , Lactancia Materna , Femenino , Liofilización , Humanos , Lactante , Leche Humana , Pasteurización
10.
J Mol Biol ; 433(10): 166956, 2021 05 14.
Artículo en Inglés | MEDLINE | ID: mdl-33775667

RESUMEN

The Covid-19 pandemic is a centenarial global catastrophe. Similar events are likely to be recurring with more frequency in the future. The inability to control the virus' impact is caused by many factors, but the lack of a technology infrastructure to detect and impede the virus at an early stage are principal shortcomings. Using phage display mutagenesis, we have generated a cohort of high performance antibody fragments (Fabs) that can be used in a sensitive point of care (POC) assay and are potent inhibitors (IC50-0.5 nM) to viral entry into cells. The POC assay is based on a split-enzyme (ß-lactamase) complementation strategy that detects virus particles at low nM levels. We have shown that this assay is equally effective for detecting other viruses like Ebola and Zika. Importantly, its components can be freeze dried and stored, but becomes fully active when rehydrated.


Asunto(s)
Anticuerpos Neutralizantes/química , Anticuerpos Neutralizantes/inmunología , Afinidad de Anticuerpos , Fragmentos Fab de Inmunoglobulinas/química , Fragmentos Fab de Inmunoglobulinas/inmunología , /aislamiento & purificación , Secuencia de Aminoácidos , Animales , Chlorocebus aethiops , Epítopos de Linfocito B/química , Epítopos de Linfocito B/inmunología , Liofilización , Prueba de Complementación Genética , Pruebas de Neutralización , Biblioteca de Péptidos , Sistemas de Atención de Punto , Glicoproteína de la Espiga del Coronavirus/química , Glicoproteína de la Espiga del Coronavirus/inmunología , Células Vero , Internalización del Virus , beta-Lactamasas/metabolismo
11.
J Vis Exp ; (168)2021 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-33645568

RESUMEN

Roots extensively interact with their soil environment but visualizing such interactions between roots and the surrounding rhizosphere is challenging. The rhizosphere chemistry of wetland plants is particularly challenging to capture because of steep oxygen gradients from the roots to the bulk soil. Here a protocol is described that effectively preserves root structure and rhizosphere chemistry of wetland plants through slam-freezing and freeze drying. Slam-freezing, where the sample is frozen between copper blocks pre-cooled with liquid nitrogen, minimizes root damage and sample distortion that can occur with flash-freezing while still minimizing chemical speciation changes. While sample distortion is still possible, the ability to obtain multiple samples quickly and with minimal cost increases the potential to obtain satisfactory samples and optimizes imaging time. The data show that this method is successful in preserving reduced arsenic species in rice roots and rhizospheres associated with iron plaques. This method can be adopted for studies of plant-soil relationships in a wide variety of wetland environments that span concentration ranges from trace-element cycling to phytoremediation applications.


Asunto(s)
Elementos , Imagenología Tridimensional/métodos , Raíces de Plantas/química , Rizosfera , Humedales , Liofilización , Oryza/anatomía & histología , Suelo/química
12.
AAPS PharmSciTech ; 22(3): 92, 2021 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-33683477

RESUMEN

Venetoclax (VX) used in the treatment of chronic lymphocytic leukemia possesses low oral bioavailability (5.4%) and undergoes first-pass metabolism. Development of a formulation to overcome its bioavailability problem can be done by using nanocrystals which has many scientific applications. Nanocrystals of VX were formulated using amalgamation of precipitation and high-pressure homogenization method, in which polyvinyl alcohol (PVA) was selected as stabilizer. Process parameters like concentration of stabilizer, homogenization pressure, number of homogenization cycle, and concentration of lyoprotectant were optimized to obtain the desired particle size for the preparation of nanocrystal formulation. HPLC methods were developed and validated in-house for determination of in vitro dissolution data and in vivo bioavailability data. Physicochemical characterization was done to determine the particle size (zeta sizer), crystalline nature (DSC and XRPD), solubility (shaker bath), and dissolution (USP type 2 apparatus). Lyophilized VX nanocrystals of size less than 350 nm showed substantial increase in saturation solubility (~20 folds) and dissolution in comparison with free VX. In vitro release study revealed that 100% dissolution was achieved in 120 min as compared to VX free base which is having less than 43.5% dissolution in 120 min. Formulations of VX remain stable for 6 months under accelerated stability conditions. In vivo pharmacokinetic data in male Sprague-Dawley rats showed (~2.02 folds) significant increase in oral bioavailability of VX formulation as compared to free drug because of rapid dissolution and absorption which makes the nanocrystal formulation a better approach for oral administration of poorly soluble drugs.


Asunto(s)
Antineoplásicos/administración & dosificación , Compuestos Bicíclicos Heterocíclicos con Puentes/administración & dosificación , Sulfonamidas/administración & dosificación , Administración Oral , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Disponibilidad Biológica , Compuestos Bicíclicos Heterocíclicos con Puentes/química , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacocinética , Cromatografía Líquida de Alta Presión , Liofilización , Masculino , Nanopartículas , Tamaño de la Partícula , Alcohol Polivinílico , Ratas , Ratas Sprague-Dawley , Solubilidad , Sulfonamidas/química , Sulfonamidas/farmacocinética
13.
AAPS PharmSciTech ; 22(3): 82, 2021 Feb 23.
Artículo en Inglés | MEDLINE | ID: mdl-33624199

RESUMEN

Current trends in the pharmaceutical industry led to a demand for more flexible manufacturing processes with smaller batch sizes. Prepackaged nested vials that can be processed as a unit were introduced into the market to fulfill this need. However, vial nests provide a different thermal environment for the vials compared to a hexagonal packaging array and could therefore influence product temperature profiles, primary drying times, and product quality attributes. Polymer caps with the possibility of vial closure inside the freeze-drying chamber were developed to remove the risks and need of a crimping process. A general concern with the use of such caps is the possibility of an increase in resistance to water vapor flow out of the vial. This case study investigated the effect of the LyoSeal® and PLASCAP® polymer caps and EZ-fill® nests on the freeze-drying process. Amorphous and partially crystalline model formulations were freeze-dried. Process data and product quality attributes were compared for regularly stoppered vials and vials with polymer caps as well as vials in a hexagonal packaging array and nested vials. The results indicated no increased resistance or impeded water vapor flow by the polymer caps. Differences in the macro- and microscopic appearances of products and a trend towards lower product temperatures were observed for the investigated nest type compared to a regular hexagonal packaging array. Consequently, the polymer caps could be used as an alternative to regular stoppers without affecting freeze-drying process data or product quality attributes, while the different thermal environment of nested vials should be considered.


Asunto(s)
Industria Farmacéutica/normas , Embalaje de Medicamentos/normas , Polímeros/normas , Desecación/métodos , Industria Farmacéutica/métodos , Embalaje de Medicamentos/métodos , Liofilización/métodos , Liofilización/normas , Temperatura
14.
Food Chem ; 350: 129188, 2021 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-33588280

RESUMEN

Hempseed meal after protein isolation (HM-PI) is a co-product obtained from hempseed. The objectives were to characterize and determine the effect of drying on HM-PI. HM-PI was produced using three drying methods: freeze (FD), vacuum oven (VOD), and oven drying (OD). HM-PI contained over 70% protein and had similar or higher level of essential amino acids than recommended values for human adults. Osborne fractionation indicated that glutelin was the most dominant fraction in HM-PI. FD HMPI has a significant lower surface hydrophobicity and higher in vitro protein digestibility than OD and VOD HM-PI. FD HM-PI demonstrated better functional properties than OD and VOD HM-PI. Pepsin-pancreatin digestion of VOD, FD and OD resulted in comparable and considerable antioxidant and anti-inflammatory properties. This is the first report on the characterization of HM-PI, a co-product of hempseed processing. HM-PI could serve as a novel food protein ingredient resulting in increase utilization of hempseed.


Asunto(s)
Cannabis/química , Fenómenos Químicos , Desecación/métodos , Resinas de Plantas/química , Antioxidantes/análisis , Liofilización , Humanos , Pancreatina/metabolismo , Pepsina A/metabolismo , Resinas de Plantas/aislamiento & purificación , Resinas de Plantas/metabolismo
15.
Int J Biol Macromol ; 172: 350-359, 2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33453258

RESUMEN

The improper management of wound exudates can expose the wound to bacterial invasion, skin maceration etc. thereby resulting in prolonged wound healing. Biopolymers are characterized by hydrophilic functional groups which when employed for the development of wound dressings promote the wound dressings capability to absorb a high amount of wound exudates. Alginate-gum acacia sponges were prepared from a combination of biopolymers such as sodium alginate and gum acacia in varying amounts with carbopol via crosslinking with 1 and 2% CaCl2. The prepared sponges were loaded with a combination of ampicillin and norfloxacin. In vitro antibacterial analysis revealed that the antibacterial activity of the loaded antibiotics was retained and the sponges were effective against gram-positive and gram-negative bacteria. The sponges displayed rapid and high absorption capability in the range of 1022-2419% at pH 5.5 simulating wound exudates, and 2268-5042% at pH 7.4 simulating blood within a period of 1-3 h. Furthermore, the whole blood clotting studies further revealed low absorbance values when compared to the control revealing the good clotting capability of the sponges. The unique features of the sponges revealed their potential application for the management of infected, high exuding and bleeding wounds.


Asunto(s)
Resinas Acrílicas/química , Alginatos/química , Antibacterianos/farmacología , Vendajes , Cloruro de Calcio/química , Goma Arábiga/química , Ampicilina/química , Ampicilina/farmacología , Antibacterianos/química , Coagulación Sanguínea/efectos de los fármacos , Liofilización/métodos , Humanos , Pruebas de Sensibilidad Microbiana , Norfloxacino/química , Norfloxacino/farmacología , Porosidad , Proteus vulgaris/efectos de los fármacos , Proteus vulgaris/crecimiento & desarrollo , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus epidermidis/efectos de los fármacos , Staphylococcus epidermidis/crecimiento & desarrollo
16.
Molecules ; 26(1)2021 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-33466395

RESUMEN

Chickpea cooking water (CCW), known as aquafaba, has potential as a replacement for egg whites due to its emulsion and foaming properties which come from the proteins and starch that leach out from chickpeas into the cooking water. High pressure (HP) processing has the ability to modify the functional characteristics of proteins. It is hypothesized that HP processing could favorably affect the functional properties of CCW proteins by influencing their structure. The objective of this study to evaluate the effect of HP treatment on the associated secondary structure, emulsion properties and thermal characteristics of CCW proteins. A central composite rotatable design is used with pressure level (227-573 MPa) and treatment time (6-24 min) as HP variables, and concentration of freeze dried CCW aquafaba powder (11-29%) as product variable, and compared to untreated CCW powder. HP improves aquafaba emulsion properties compared to control sample. HP reduces protein aggregates by 33.3%, while ß-sheets decreases by 4.2-87.6% in which both correlated to increasing protein digestibility. α-helices drops by 50%. It affects the intensity of some HP treated samples, but not the trend of bands in most of them. HP treatment decreases Td and enthalpy because of increasing the degree of denaturation.


Asunto(s)
Cicer/química , Culinaria/métodos , Manipulación de Alimentos/métodos , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Presión , Liofilización , Proteínas de Plantas/análisis , Agua
17.
Molecules ; 26(3)2021 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-33498977

RESUMEN

There is a significant indication of the beneficial health effects of fruit rich diets. Fruits of native plant species have noticeably different phytochemicals and bioactive effects. The aim of this work was to characterize and compare the constituents of jabuticaba (Myrciaria jaboticaba, MJ), jamun-berry (Syzygium cumini, SC), and malay-apple (Syzygium malaccense, SM) extracts and their influence on antioxidant activity in vitro and antiproliferative effects on human colon adenocarcinoma cells. According to the results, dried peel powders (DP) have a high anthocyanin content, phenolic compounds, and antioxidant activity when compared to freeze dried extracts (FD). M. jaboticaba dried peel powder extract had a higher total anthocyanin and phenolic compounds content (802.90 ± 1.93 and 2152.92 ± 43.95 mg/100 g, respectively). A reduction in cell viability of HT-29 cells after treatment with M. jaboticaba extracts (DP-MJ and FD-MJ) was observed via MTT assay. Flow cytometry showed that the treatment with the anthocyanin-rich extracts from MJ, SC, and SM had an inhibitory impact on cell development due to G2/M arrest and caused a rise in apoptotic cells in relation to the control group. The findings of this study highlight the potential of peel powders from Myrtaceae fruits as an important source of natural antioxidants and a protective effect against colon adenocarcinoma.


Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Antocianinas/uso terapéutico , Neoplasias del Colon/tratamiento farmacológico , Myrtaceae/química , Fitoterapia , Adenocarcinoma/patología , Antocianinas/análisis , Antioxidantes/análisis , Antioxidantes/uso terapéutico , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Neoplasias del Colon/patología , Liofilización , Frutas/química , Células HT29 , Humanos , Fenoles/análisis , Fenoles/uso terapéutico , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Syzygium/química
18.
Biochemistry ; 60(2): 152-159, 2021 01 19.
Artículo en Inglés | MEDLINE | ID: mdl-33400518

RESUMEN

Water is key to protein structure and stability, yet the relationship between protein-water interactions and structure is poorly understood, in part because there are few techniques that permit the study of dehydrated protein structure at high resolution. Here, we describe liquid-observed vapor exchange (LOVE) NMR, a solution NMR-based method that provides residue-level information about the structure of dehydrated proteins. Using the model protein GB1, we show that LOVE NMR measurements reflect the fraction of the dried protein population trapped in a conformation where a given residue is protected from exchange with D2O vapor. Comparisons to solution hydrogen-deuterium exchange data affirm that the dried protein structure is strongly influenced by local solution stability and that the mechanism of dehydration protection exerted by the widely used protectant trehalose differs from its mechanism of stabilization in solution. Our results highlight the need for refined models of cosolute-mediated dehydration protection and demonstrate the ability of LOVE NMR to inform such models.


Asunto(s)
Aminoácidos/química , Proteínas Bacterianas/química , Deuterio/química , Hidrógeno/química , Espectroscopía de Resonancia Magnética/métodos , Streptococcaceae/metabolismo , Medición de Intercambio de Deuterio , Liofilización , Conformación Proteica
19.
Nat Commun ; 12(1): 304, 2021 01 12.
Artículo en Inglés | MEDLINE | ID: mdl-33436631

RESUMEN

Skeletal muscle conveys several of the health-promoting effects of exercise; yet the underlying mechanisms are not fully elucidated. Studying skeletal muscle is challenging due to its different fiber types and the presence of non-muscle cells. This can be circumvented by isolation of single muscle fibers. Here, we develop a workflow enabling proteomics analysis of pools of isolated muscle fibers from freeze-dried human muscle biopsies. We identify more than 4000 proteins in slow- and fast-twitch muscle fibers. Exercise training alters expression of 237 and 172 proteins in slow- and fast-twitch muscle fibers, respectively. Interestingly, expression levels of secreted proteins and proteins involved in transcription, mitochondrial metabolism, Ca2+ signaling, and fat and glucose metabolism adapts to training in a fiber type-specific manner. Our data provide a resource to elucidate molecular mechanisms underlying muscle function and health, and our workflow allows fiber type-specific proteomic analyses of snap-frozen non-embedded human muscle biopsies.


Asunto(s)
Adaptación Fisiológica , Ejercicio Físico , Liofilización , Músculo Esquelético/metabolismo , Músculo Esquelético/fisiología , Proteómica , Biomarcadores/metabolismo , Biopsia , Glucosa/metabolismo , Humanos , Mitocondrias/metabolismo , Fibras Musculares de Contracción Rápida/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares de Contracción Lenta/metabolismo , Análisis de Componente Principal , Proteoma/metabolismo
20.
AAPS PharmSciTech ; 22(2): 57, 2021 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-33502633

RESUMEN

Recent advances in molded vial manufacturing enabled manufacturers to use a new manufacturing technique to achieve superior homogeneity of the vial wall thickness. This study evaluated the influence of the different manufacturing techniques of molded vials and glass compositions on vial heat transfer in freeze-drying. Additionally, the influence of using empty vials as thermal shielding on thermal characteristics of edge and center vials was investigated. The vial heat transfer coefficient Kv was determined gravimetrically for multiple vial systems. The results showed superior heat transfer characteristics of the novel manufacturing technique as well as differences in heat transfer for the different glass compositions. Empty vials on the outside of the array did not influence center vial Kv values compared to a full array. The direct contact area and vial bottom curvature and their correlation to heat transfer parameters were analyzed across multiple vial systems. A new approach based on light microscopy to describe the vial bottom curvature more accurately was described. The presented results for the contact area allowed for an approximation of the pressure-independent heat transfer parameter KC. The results for the vial bottom curvature showed a great correlation to the pressure-dependent heat transfer parameter KD. Overall, the results highlighted how a thorough geometrical characterization of vials with known heat transfer characteristics could be used to predict thermal characteristics of new vial systems as an alternative to a time-consuming gravimetric Kv determination. Primary drying times were simulated to show the influence of Kv on drying performance.


Asunto(s)
Embalaje de Medicamentos/métodos , Liofilización/métodos , Tecnología Farmacéutica/métodos , Vidrio/química , Calor
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