Construction and evaluation of novel self-assembled nanoparticles of Herpetospermum caudigerum Wall / 药学学报

It has become an industry consensus that self-assembled nanoparticles (SAN) are formed by molecular recognition of chemical components in traditional Chinese medicine during the decoction process. The insoluble components in the decoction are mostly in the form of nanoparticles, which can improve the problem of poor water solubility. However, the transfer rate of these insoluble components in the decoction is still very low, which limits the efficacy of the drug. This study aimed to refine the traditional decoction self-assembly phenomenon. The self-assembled nanoparticles were constructed by micro-precipitation method (MP-SAN), and characterized by particle size, zeta potential, stability index and morphology. The formation of MP-SAN and alterations in related physicochemical properties were evaluated using modern spectroscopic and thermal analysis techniques. The quality value transmitting pattern of lignan components within the MP-SAN was assessed via high performance liquid chromatography (HPLC). The MP-SAN showed sphere-like structure with uniform morphology, particle size of (245.3 ± 3.2) nm, polydispersity index (PDI) of (0.13 ± 0.03), zeta potential of (-48.9 ± 5.9) mV and stability index (SI) of (86.05% ± 2.27%). Comprehensive analyses using ultraviolet visible spectroscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, and other techniques confirmed molecular recognition between the decoction and ethanol extraction, leading to electron rearrangement under the influence of non-covalent bonding. This resulted in the formation of nanoparticles possessing superior thermal stability. As determined by HPLC, the encapsulation rates of the index components in the MP-SAN were all greater than 75% (dehydrodiconiferyl alcohol: 77.00%; herpetolide A: 78.57%; herpetrione: 94.53%), and the transfer rates were all higher than 65% (dehydrodiconiferyl alcohol: 96.01%; herpetolide A: 67.86%; herpetrione: 65.55%), which were 1.34, 1.38 and 4.81 times compared with those of the traditional decoction. In summary, this study successfully constructed the MP-SAN based on micro-precipitation method to achieve high transfer rate and high encapsulation rate of insoluble components in docoction, which provides a pharmaceutics idea for the efficient utilization of pharmacodynamic substance basis of traditional Chinese medicine.

Optimization of ethanol precipitation process of Nauclea officinalis extract based on concept of quality by design(QbD) / 中国中药杂志

The ethanol precipitation process of Nauclea officinalis extract was optimized based on the concept of quality by design(QbD). Single factor tests were carried out to determine the levels of test factors. The ethanol volume fraction, pre-ethanol precipitation drug concentration, and ethanol precipitation time were taken as critical process parameters(CPPs). With the comprehensive scores of strictosamide transfer rate and solid removal rate as the critical quality attributes(CQAs), Box-Behnken design was employed to establish the mathematical models and space design between CPPs and CQAs, and the obtained optimal operating space was validated. The optimal operating space included ethanol volume fraction of 65%-70%, pre-ethanol precipitation drug concentration of 22-27 mg·mL~(-1), and ethanol precipitation time of 12 h. Based on the concept of QbD, this study adopted the design space to optimize the ethanol precipitation process of N. officinalis extract, which provided a reliable theoretical basis for the quality control in the production process of N. officinalis preparations. Moroever, this study provided a reference value for guiding the research and industrial production of traditional Chinese medicines.

Quantification of antigen of Mycoplasma capricolum subsp. capripneumoniae by optical assay / 生物工程学报

Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.

Synthesis of iron nanoparticles, its characterization and anti-microbial activity assessment

We report the efficacy of the Iron nanoparticles (IONPs) and assessed two different approaches for the synthesis of IONPs i.e. Polyol and co-precipitation method and further, evaluate their antimicrobial properties. Ferrous sulphate heptahydrate salts were reduced with ethylene glycol to obtain IONP and Fe+2 and Fe+3 co-precipitation reaction was performed with KOH at optimum heating. Further, synthesized (IONPs) were characterized by hydrodynamic radii measurement done by DLS clearly indicating the size of IONPs is 79.75nm in polyol based and 135.1 nm in co-precipitation method. The biological efficacy in terms of antimicrobial activity was assessed by the Kirby Bauer method, applied for both Gram-positive and Gram-negative bacteria such as Staphylococcus aureus and Escherichia coli, respectively. The ZOI values i.e. Zone of inhibition diameter was found to be clearly visible in both S. aureus and E. coli, indicating bactericidal activity. Further growth kinetics studies and bacterial genotoxicity was also assessed. Hence, IONPs synthesized are proposed to have great potential as an antibacterial agent and can be used in drug delivery.

Kyasanur forest disease and climatic attributes in India

Background & objectives: In India, Kyasanur Forest Disease has been reported from the states of Karnataka, Kerala, Goa, and Maharashtra. The relationship between climatic factors and transmission of KFD remains untouched, therefore, the present study was undertaken. Methods: Based on the occurrence of cases, Shivamogga district (Karnataka) and Wayanad district in Kerala and northern Goa (Goa state) were selected for the study. Data on the incidence of KFD and climate factors were collected from concerned authorities. To determine the relationship between dependent and independent variables, spearman’s correlation was calculated for monthly as well as with lag months. Results: KFD cases and temperature (°C) were found significantly correlated up to 1 months’ lag period (p<0.05) while with precipitation relationship was found negatively significant for 0-3 months’ lag. The range of suitable temperature for KFD in Shivamogga, Goa and Wayanad was found as 20-31°C, 25-29°C and 27-31°C respectively. The cumulative precipitation during transmission months (November–May) ranged from <150-500mm, while in non-transmission months (June-October) from >1100-2400mm. Interpretation & conclusion: The analysis of three sites revealed that with the increase in temperature, the intensity of KFD transmission decreases as corroborated by the seasonal fluctuations in Shivamogga, Goa and Wayanad. High precipitation from June to October rovides suitable ecology to tick vector and sets in transmission season from November to May when cumulative precipitation is <500 mm.

A case report of macro-AST with isolated elevated aspartate aminotransferase in a child / 中华检验医学杂志

We need to consider the macro-AST when the elevated AST activity cannot be explained. A 3-year-old child was found to have an increase in serum AST activity, but no obvious abnormality be found ofter examination. The PEG precipitation assay showed that the activity was 98.7%, which was diagnosed as macro-AST.

Advances of enzymes related to microbial cement / 生物工程学报

Microbial induced calcium carbonate precipitation (MICP) refers to the natural biological process of calcium carbonate precipitation induced by microbial metabolism in its surrounding environment. Based on the principles of MICP, microbial cement has been developed and has received widespread attention in the field of biology, civil engineering, and environment owing to the merits of environmental friendliness and economic competence. Urease and carbonic anhydrase are the key enzymes closely related to microbial cement. This review summarizes the genes, protein structures, regulatory mechanisms, engineering strains and mutual synergistic relationship of these two enzymes. The application of bioinformatics and synthetic biology is expected to develop biocement with a wide range of environmental adaptability and high performance, and will bring the MICP research to a new height.

Optimization of formulation of paclitaxel nanosuspension encapsulated by erythrocyte membrane based on Box-Behnken method / 中国中药杂志

In view of the longevity and innate immune escape of red blood cells, this study designed the red blood cell membrane-coated paclitaxel nanosuspension [RBC-(PTX)NS] and investigated its physicochemical properties and antitumor effect in vitro. Paclitaxel nanosuspension [(PTX)NS] was prepared by ultrasonic precipitation and then RBC-(PTX)NS by ultrasonic coating. The formulation of(PTX)NS was optimized with Box-Behnken method and indexes of particle diameter, zeta potential, and stability. The morphology, particle diameter, stability, in vitro dissolution, and antitumor effect of(PTX)NS and RBC-(PTX)NS were characterized. The results showed that the particle diameter and zeta potential were(129.38±0.92) nm and(-22.41±0.48) mV, respectively, for the optimized(PTX)NS, while(142.5±0.68) nm and(-29.85±0.53) mV, respectively, for RBC-(PTX)NS. Under the transmission electron microscope,(PTX)NS was spherical and RBC-(PTX)NS had obvious core-shell structure. RBC-(PTX)NS remained stable for 5 days at 4 ℃. The in vitro dissolution test demonstrated that the cumulative release rate of RBC-(PTX)NS reached 79% within 20 min, which was significantly higher than that(25%) of(PTX)NS(P<0.05). As evidenced by MTT assay, RBC-(PTX)NS highly inhibited the proliferation of HepG2 cells in a dose-dependent manner. The cell membrane-coated nano-preparation preparation method is simple and reproducible. It improves the solubility of PTX and endows RBC-(PTX)NS with higher stability and stronger cytotoxicity. Thus, it is a new method for the delivery of PTX via nanocrystallization.

Two-Step Purification of L-Asparaginase from Acrylaway® L

Abstract L-Asparaginase (L-ASNase) is a biopharmaceutical used for acute lymphoblastic leukaemia (ALL) treatment, dramatically increasing the patients' chance of cure. However, its production and distribution in developing countries were disrupted because of its low profitability, which caused great concern among patients. This study evaluates the feasibility of combining fractional precipitation and aqueous two-phase systems (ATPS) to purify L-ASNase from a low-grade product, commercially known as Acrylaway® L. The ATPS purification results were not particularly expressive compared to the two-step purification process composed of ethanol precipitation and gel filtration, which was able to recover the target molecule with a purification factor over 5 fold. Thus, we studied a purification process capable of manufacturing pharmaceutical grade L-ASNase from a commercially available low-grade raw material; however, improvements regarding its throughput must be achieved, and high purity is the first step to apply it as a new biopharmaceutical product. The proposed process could pose as a short-time solution to mitigate its shortage while a cost-effective production plant is being developed.

Efavirenz dissolution enhancement V - A combined top down/bottom up approach on nanocrystals formulation

Abstract Efavirenz is one of the most commonly used drugs in HIV therapy. However the low water solubility tends to result in low bioavailability. Drug nanocrystals, should enhance the dissolution and consequently bioavailability. The aim of the present study was to obtain EFV nanocrystals prepared by an antisolvent technique and to further observe possible effect, on the resulting material, due to altering crystallization parameters. A solution containing EFV and a suitable solvent was added to an aqueous solution of particle stabilizers, under high shear agitation. Experimental conditions such as solvent/antisolvent ratio; drug load; solvent supersaturation; change of stabilizer; addition of milling step and solvents of different polarities were evaluated. Suspensions were characterized by particle size and zeta potential. After freeze- dried and the resulting powder was characterized by PXRD, infrared spectroscopy and SEM. Also dissolution profiles were obtained. Many alterations were not effective for enhancing EFV dissolution; some changes did not even produced nanosuspensions while other generated a different solid phase from the polymorph of raw material. Nevertheless reducing EFV load produced enhancement on dissolution profile. The most important modification was adding a milling step after precipitation. The resulting suspension was more uniform and the powder presented grater enhancement of dissolution efficacy.

Análisis comparativo de casos de suicidio por ahorcadura, precipitación e intoxicación atendidos en la Morgue Judicial en la provincia de Ancón, Panamá de marzo del 2019 a febrero de 2020 y marzo del 2020 a febrero de 2021 / Comparative analysis of suicide by hanging, precipitation and intoxication cases attended at the Judicial Morgue in the province of Ancón, Panama from March 2019 to February 2020 and March 2020 to February 2021

Resumen Introducción: La crisis por la Covid-19, en muchas maneras, podría exacerbar ciertas dimensiones del proceso suicida a través de su impacto psicológico, social, económico o, incluso, biológico. El estudio comparará los casos de suicidio por ahorcadura, precipitación e intoxicación atendidos en la morgue judicial de la provincia de Panamá, entre marzo del 2019 a febrero del 2020 y marzo del 2020 a febrero del 2021. Material y métodos: Se trata de un estudio descriptivo, transversal y retrospectivo; cuya población de análisis comprende todas las autopsias médico-legales realizadas en la Morgue Judicial de Panamá por casos de ahorcadura, precipitación e intoxicación, de marzo del 2019 a febrero del 2021. Resultados: En el estudio se registró un total de 117 casos. De estos, 73 practicados de marzo del 2019 a febrero del 2020, de los cuales el 78 % (n=57) correspondió al sexo masculino, con una media de 41 años, y el 22 % (n=16) al sexo femenino, con una media de 36 años; mientras que, en el lapso de marzo del 2020 a febrero del 2021 la muestra fue de 44 casos, lo que representó el 93 % (n=41), de sexo masculino, con una media de 42 años y el 7 % (n=3) de sexo femenino, con una media de 47 años. Conclusión: Durante el año posterior al inicio de la pandemia y la aplicación de las medidas de cuarentena en la provincia de Panamá hubo una disminución de la frecuencia de los casos de suicidios por ahorcadura e intoxicación en comparación con el año previo y un aumento en cuanto a los casos por precipitación.

Abstract Introduction: The Covid-19 crisis in many ways could exacerbate certain dimensions of the suicidal process through its psychological, social, economic or even biological impact. In this study we will compare the cases of suicide by hanging, precipitation and intoxication carried out in the Judicial Morgue of Ancon from March 2019 to February 2020 and from March 2020 to February 2021. Materials and methods: This is a descriptive, cross-sectional and retrospective study; the population studied will be all the medicolegal autopsies performed in the Judicial Morgue of Panama, due to hanging, precipitation and intoxication in the period from March 2019 to February 2021. Results: A total of 117 cases were recorded in this period studied; 73 cases from March 2019 to February 2020 of which 78% (n=57) corresponded to the male sex, with an average of 41 years and 22% (n=16) to the female sex, with an average of 36; while, in the period from March 2020 to February 2021, of the sample of 44 cases 93% (n=41) corresponded to the male sex, with an average of 42 and 7% (n=3) to the female sex, with an average of 47 years. Conclusión: During the year after the beginning of the pandemic and the quarantine measures in the Province of Panama, there was a decrease in the frequency of suicide cases by hanging, precipitation and intoxication compared to the previous year.

Síntesis del hidróxido de níquel (II) con disolución multicomponente de la lixiviación de sulfuro de la tecnología Caron / Nickel hydroxide (II) synthesis with sulfide leaching multi-element solution from Caron technology / Síntese de hidróxido de níquel (II) com solução de lixiviação de sulfetos multicomponentes da Caron technology

Resumen Se determinaron las condiciones más favorables para la síntesis de hidróxido de níquel (II) por precipitación química a partir de la disolución lixiviada del sulfuro de níquel de la tecnología Caron. La disolución lixiviada de fuerza iónica igual a 4,6 mol-L-1 fue tratada con hidróxidos de amonio y sodio. Se calcularon las propiedades termodinámicas como entalpía, entropía y energía libre y se construyeron diagramas de zona de predominancia. Se demostró la conveniencia de realizar el proceso a 303 K, pH entre 11 y13 y concentración de amoníaco de 0,5 a 1,0 mol•L-1. En estas condiciones fueron obtenidas altas eficiencias de precipitación de los cationes metálicos de la disolución inicial. El sólido precipitado se caracterizó mediante espectrometría de absorción atómica y difracción de rayos X. Se obtuvo un beta-hidróxido de níquel (II) con la siguiente composición química: 58,5 ≤ Ni ≤ 59,1%, 1,7 ≤ Co ≤ 2,1%, 0,5 ≤ Fe ≤ 0,6%, Cu, Zn, Mn y Mg ≤ 0,2%.

Abstract The most favorable conditions for the synthesis of nickel (II) hydroxide were determined by chemical precipitation, from the leached solution of nickel sulfide from the Caron technology. The leached solution with an ionic strength equal to 4.6 mol-L-1 was treated with ammonium and sodium hydroxides. Thermodynamic properties such as enthalpy, entropy, and free energy were calculated and predominance zone diagrams were constructed. The convenience of performing the process at 303 K, pH between 11 and 13, and ammonia concentration of 0.5 to 1.0 mol•L-1 was demonstrated. Under these conditions, high precipitation efficiencies of the metal cations from the initial solution were obtained. The precipitated solid was characterized by atomic absorption spectrometry and X-ray diffraction. Nickel (II) beta-hydroxide with the following chemical composition was obtained: 58.5 ≤ Ni ≤ 59.1%, 1.7 ≤ Co ≤ 2.1%, 0.5 ≤ Fe ≤ 0.6%, Cu, Zn, Mn and Mg ≤ 0.2%.

Resumo As condições mais favoráveis para a síntese do hidróxido de níquel (II) foram determinadas por precipitação química, a partir da solução lixiviada de sulfeto de níquel da tecnologia Caron. A solução lixiviada de força iônica igual a 4,6 mol-L-1, foi tratada com hidróxidos de amônio e sódio. Foram calculadas propriedades termodinâmicas, como entalpia, entropia e energia livre, e diagramas de zonas de predominância foram construídos. Foi demonstrada a conveniência de realizar o processo a 303 K, pH entre 11 e 13 e concentração de amônia de 0,5 a 1,0 mol•L-1. Nessas condições, foram obtidas altas eficiências de precipitação dos cátions metálicos da solução inicial. O sólido precipitado foi caracterizado por espectrometria de absorção atômica e difração de raios-x. Foi obtido beta-hidróxido de níquel (II) com a seguinte composição química: 58,5 ≤ Ni ≤ 59,1%, 1,7 ≤ Co ≤ 2,1%, 0,5 ≤ Fe ≤ 0,6%, Cu , Zn, Mn e Mg ≤ 0,2%.

Caracterização da seca histórica da década de 2010 na Bacia do Rio Paraíba do Sul, Estado de São Paulo, Brasil / Characterization of the 2010-decade historical drought in Paraiba do Sul River Basin, São Paulo State, Brazil

RESUMO A Bacia Hidrográfica do Rio Paraíba do Sul foi afetada por uma das secas mais severas de sua história, durante os anos de 2013 a 2015, que resultou em várias consequências para o gerenciamento de seus recursos hídricos. O objetivo deste artigo foi contribuir com o entendimento desse evento histórico, no trecho Paulista da Bacia do Rio Paraíba do Sul, por meio do cálculo e da análise do índice padronizado de precipitação. Esse índice foi calculado nas escalas de 1, 3, 6 e 12 meses, para 22 estações pluviométricas localizadas na área de estudo. Os resultados demonstraram que a seca ocorrida entre 2013 e 2014 foi uma das mais severas já registradas, e que o verão de 2014 foi o período crítico no que diz respeito à redução da chuva regional. Essa seca impactou o armazenamento e a capacidade de regularização do principal reservatório da região, o Reservatório de Paraibuna. Embora o pico da crise hídrica tenha ocorrido, principalmente, em razão da seca de 2013/2014, verificou-se que ele foi influenciado pelo efeito cumulativo de uma seca anterior, ocorrida desde o início do ano de 2011. Esse resultado reforça a importância do planejamento plurianual da operação do Sistema Hidráulico da Bacia do Rio Paraíba do Sul.

ABSTRACT The Paraiba do Sul River Basin was affected by one of the most severe drought periods in its recent history, during the years 2013 to 2015, which resulted in several consequences for the management of its water resources. This article aims to contribute to the understanding of this historical event in the São Paulo Stretch of the Paraiba do Sul River Basin, through the calculation and analysis of the standardized precipitation index (SPI). SPI was calculated for the time scales of 1, 3, 6, and 12 months, for 22 rainfall stations located in the study area. The results showed that the drought occurred between 2013 and 2014 was one of the most severe ever recorded, and that the summer of 2014 was the critical period in terms of reduction of regional rainfall. This drought impacted the storage and regularization capacity of the main reservoir of the region, the Paraibuna Reservoir. Although the peak of the water crisis occurred mainly due to the 2013/2014 drought, it was found that it was influenced by the cumulative effect of a previous dry period, that occurred since the beginning of the year of 2011. This result reinforces the importance of multiannual planning for the operation of the Paraíba do Sul River Basin Hydraulic System.

Cambios climáticos proyectados de modelos CMIP5 en La Cruz, Guanacaste, Costa Rica

Introducción: La Cruz está localizada en el noroeste de Costa Rica, y está caracterizada por una estación seca bien definida y un clima tropical seco. Este cantón en los últimos 45 años ha reportado impactos de eventos hidrometeorológicos, en particular los húmedos como las inundaciones que han afectado viviendas, calles y cultivos, pero también algunas sequías han perjudicado a la población y sus actividades. Objetivo: El objetivo de este artículo es proveer información acerca de posibles cambios climáticos durante el siglo XXI asociados con un escenario pesimista de emisiones en el cantón de La Cruz en Guanacaste, Costa Rica. Métodos: Se utilizó la base de datos DesInventar para identificar los impactos y agruparlos según su mes de ocurrencia para relacionarlo con el régimen de precipitación. Simulaciones cambiadas de escala del Proyecto de Intercomparación de Modelos Climáticos 5 (CMIP5) fueron estados para simular cambios pasados y futuros (1979-2099) en temperatura y precipitación. Adicionalmente, un modelo hidrológico fue usado para calcular las proyecciones de escorrentía usando los datos meteorológicos. Resultados: Los resultados mostraron que un futuro más seco y caliente será característico del clima del siglo XXI, especialmente después del 2040. La escorrentía se espera que será reducida significativamente siguiendo las tendencias meteorológicas. Conclusiones: Algunos de los impactos más recurrentes históricamente son las inundaciones, daños a viviendas, calles y cultivos, sin embargo, también se han registrado algunas muertes en la zona. Se espera que una aridez mayor estará presente en el futuro, afectando sistemas humanos y ambientales.

Introduction: La Cruz is located in northwestern Costa Rica, and it is characterized by a dry tropical climate with a well-defined dry season. This municipality has reported many impacts of hydrometeorological events during the last 45 years, mostly associated with wet events such as floods that affected roads, housing, and crops; as well as some droughts that have impacted the population and their livelihoods. Objective: The objective of this article is to provide information regarding possible future climate changes during the 21st century associated with a pessimistic emission scenario in the municipality of La Cruz in Guanacaste, Costa Rica. Methods: We used DesInventar database to identify the hydrometeorological impacts in La Cruz and classified them by month. Downscaled simulations from the Climate Model Intercomparison Project 5 (CMIP5) were used to simulate past and future changes (1979-2099) in temperature and precipitation. In addition, a hydrological model was used to calculate runoff projections from meteorological data. Results: Floods, and housing, roads and agricultural damages are the most recurrent impacts. In addition, some human deaths are reported during this time. Results show that a drier and warmer future will be characteristic of the climate in La Cruz in the 21st century, especially after 2040. Runoff is also expected to be reduced significantly following the meteorological trends. Conclusions: It is expected that higher aridity will be present in the future, affecting human and environmental systems.

Spatial heterogeneity and physical structure basis of antibacterial activity of Ma-Xing-Shi-Gan Decoction / 药学学报

Ma-Xing-Shi-Gan Decoction is a classic prescription. However, the interaction among multiple components of the decoction and the change of phase state are not clear. Moreover, the relationship between the physical phase state aggregated by multiple components and the efficacy still needs to be studied. In this study, we monitored the particle size changes of Ma-Xing-Shi-Gan Decoction in real time. Then we isolated different phase states by centrifugation, analyzed their composition distribution and tested their antibacterial activity. We added chemical interference agents to investigate the interaction of multi-component physical phase states accompanied by the observation of particle size change and morphology. We also studied the correlation between antibacterial activity and physical structure of phase states. The results showed during boiling process the degree of hybridization of particles was decreased and the particle size distribution was narrowed and stabilized at 170 nm. The distribution of organic and inorganic components was heterogeneous among different phase states. S-13500, supernatant isolated by 13 500 ×g centrifugation, constituted by ephedrine, amygdalin, glycyrrhizic acid and inorganic components Ca, K, Mg, etc., had the strongest antibacterial activity. The molecular interaction force in the active physical phase state was mainly hydrophobic and hydrogen bond. The destruction of the interaction force will lead to the change of phase structure and the decrease of antibacterial activity in vitro and in vivo. This study confirms that, in the boiling process of the Ma-Xing-Shi-Gan Decoction, the chemical components interweave and interact to form new physical phase states, leading to heterogeneous distribution of components. The antimicrobial activity of the active phase depends on both chemical composition and physical structure, which provides a direct evidence for the physical basis of the efficacy of traditional Chinese medicine.

Comparison of methods for enriching urine proteins / 生物工程学报

The abundance of proteins in human urine is low and easily to be masked by high-abundance proteins during mass spectrometry analysis. Development of efficient and highly selective enrichment methods is therefore a prerequisite for achieving deep coverage of urine protein markers. Notably, different experimental methods would affect the urine protein enrichment efficacy and the coverage of urine proteome. In this study, ultrafiltration, nitrocellulose membrane enrichment and saturated ammonium sulfate precipitation were used to process 10 mL urine samples from five healthy volunteers and five bladder cancer patients. The urine proteins were enriched and separate by SDS-PAGE to compare the purification efficiency of different methods. Moreover, the peptide identification effects of different purification methods were analyzed by mass spectrometry to determine the best method for enriching urine protein histones. Saturated ammonium sulfate precipitation method outperformed the ultrafiltration and the nitrocellulose membrane enrichment methods in terms of the protein enrichment efficacy and quality. The interference of highly abundant albumin was reduced, whereas the amount of low-abundance protein was increased, and the sensitivity of mass spectrometry identification was increased. The saturated ammonium sulfate precipitation method may be applied for large-scale urine processing for screening clinical diagnostic markers through proteomics.

Delivery strategies of amphotericin B for invasive fungal infections

Invasive fungal infections (IFIs) represent a growing public concern for clinicians to manage in many medical settings, with substantial associated morbidities and mortalities. Among many current therapeutic options for the treatment of IFIs, amphotericin B (AmB) is the most frequently used drug. AmB is considered as a first-line drug in the clinic that has strong antifungal activity and less resistance. In this review, we summarized the most promising research efforts on nanocarriers for AmB delivery and highlighted their efficacy and safety for treating IFIs. We have also discussed the mechanism of actions of AmB, rationale for treating IFIs, and recent advances in formulating AmB for clinical use. Finally, this review discusses some practical considerations and provides recommendations for future studies in applying AmB for combating IFIs.

Reactores Bioquímicos Pasivos: Una alternativa biotecnológica para la remediación de drenajes ácidos de mina / Biochemical passive reactors: a biotechnological alternative to remediation of acid mine drainage

RESUMEN El Drenaje ácido de mina (DAM) es actualmente el principal contaminante de las regiones mineras. Los reactores bioquímicos pasivos son una tecnología sostenible fácil de instalar que utiliza desechos agroindustriales de la región y puede operar en áreas remotas con poco mantenimiento. Además, son una tecnología limpia que involucra bioprocesos, reacciones químicas y precipitación de metales, minimizando el impacto de los vertimientos ácidos sobre suelos y cuerpos de aguas. Los reactores bioquímicos pasivos son columnas empacadas con una "mezcla reactiva" conformada por materiales orgánicos, inorgánicos y un inóculo microbiano. En esta mezcla se remedia el DAM por medio de procesos fisicoquímicos como la adsorción, precipitación, coprecipitación de los metales y de la reducción del sulfato a sulfuro, mientras se incrementa el pH y la alcalinidad. Con el fin de brindar información reciente, así como las necesidades de investigación en el tema, este documento presenta una revisión de literatura sobre la generación química y biológica de los DAM, así como su remedición utilizando reactores bioquímicos pasivos. El conocimiento de los conceptos básicos de estos procesos es extremadamente útil para evaluar las posibles aplicaciones, beneficios y limitaciones de estos sistemas de tratamiento utilizados por la biotecnología durante la biorremediación de efluentes mineros.

ABSTRACT Acid Mine Drainage (AMD) is currently the main pollutant in mining areas. Passive biochemical reactors are a sustainable technology easy to install using agro-industry waste from the mining region and operating in remote locations. Besides, bioreactors are clean technology that involves bioprocesses, chemical reactions, and metal precipitation, minimizing the impact of AMD on soils and fresh water sources. The passive biochemical reactors are columns packed with a "reactive mixture" consisting of organic, inorganic materials and a microbial inoculum. In this reactive mixture, AMD is remediated through physicochemical processes such as metals adsorption, precipitation, and co-precipitation, as well as, the reduction of sulfate to sulfur, while pH and alkalinity are in-creased. To provide recent information and research needs in the subject, this document presents a review of the literature about the chemical and biological generation of AMD and its remediation using passive biochemical reactors. The knowledge of the basic concepts of these processes is extremely useful to evaluate the possible applications, benefits and limitations of these treatment systems used by biotechnology during the bioremediation of mining effluents.

Production fields and physiological quality of Panicum maximum jacq. cv. mombasa seeds / Campos de produção e qualidade fisiológica de sementes de Panicum maximum jacq. cv. mombasa

The physiological quality of seeds is influenced by the climatic factors of production fields. The identification of the best conditions of P. maximum cv. Mombasa production fields allows the improvement of the seed sector. The aim of this work was to identify which climatic conditions of production fields can affect the physiological quality of P. maximum cv. Mombasa seeds. Nineteen plots from states of São Paulo (six from municipalities of Auriflama and three from Guzolândia) and Goiás (eight from Quirinópolis and two from Serranópolis), were collected by soil sweeping. The following parameters were evaluated: water content, germination rate, first germination count and germination rate index, seedling emergence in sand and field. Completely randomized design was used for all variables, with the exception of seedling emergence in field, since this variable required block design. Means were compared by the Scott-Knott test, at 5% probability. For the identification of the influence of climatic conditions on the physiological quality of seeds, multivariate statistical analysis was applied through Group and Principal Component Analysis. Production fields of Panicum maximum cv. Mombasa seeds presenting maximum temperatures above 32 ºC at flowering and natural fall stages produce seeds of low physiological quality. Production fields in which precipitation and high temperatures occur during natural fall and harvest of P. maximum cv. Mombasa seeds are not favorable to the production of seeds with high physiological quality.

A qualidade fisiológica das sementes é influenciada pelos fatores climáticas doscampos de produção. A identificação das condições dos melhores campos de produção de P. maximum cv. Mombaça permite aprimoramento do setor de sementes. O objetivo desse trabalho foi identificar quais condições climáticas dos campos de produção podem afetar a qualidade fisiológica das sementes de P. maximum cv. Mombaça. Foram avaliados dezenove lotes procedentes do estado de São Paulo (seis de Auriflama e três de Guzolândia) e Goiás (oito de Quirinópolis e dois de Serranópolis), colhidos por varredura do solo. Foram avaliados por meio dos seguintes parâmetros: teor de água, germinação, primeira contagem de germinação e índice de velocidade de germinação, emergência de plântulas em areia e em campo. Adotou-se delineamento inteiramente casualizado para todas as variáveis, com exceção da emergência de plântulas em campo, pois neste adotou-se delineamento em blocos. As médias foram comparadas pelo teste Scott-Knott, a 5% de probabilidade. Para a identificação da influência das condições climáticas na qualidade fisiológica das sementes aplicou-se análise estatística multivariada por meio de Análise de Agrupamento e Componentes Principais. Campos de produção de sementes de Panicum maximum cv. Mombaça que apresentam temperaturas máximas superiores a 32 ºC nas épocasde florescimento e degrana produzem sementes de baixa qualidade fisiológica. Campos de produção em que ocorram precipitações e altas temperaturas durante a degrana e à colheita de sementes de P.maximum cv. Mombaça não são favoráveis a produção de sementes com alta qualidade fisiológica.

Comparison of protein precipitation methods for sample preparation prior to proteomic analysis of Chinese hamster ovary cell homogenates

BACKGROUND: Chinese hamster ovary (CHO) cells are the workhorse for obtaining recombinant proteins. Proteomic studies of these cells intend to understand cell biology and obtain more productive and robust cell lines for therapeutic protein production in the pharmaceutical industry. Because of the great importance of precipitation methods for the processing of samples in proteomics, the acetone, methanol-chloroform (M/C), and trichloroacetic acid (TCA)-acetone protocols were compared for CHO cells in terms of protein recovery, band pattern resolution, and presence on SDS-PAGE. RESULTS: Higher recovery and similar band profile with cellular homogenates were obtained using acetone precipitation with ultrasonic bath cycles (104.18 ± 2.67%) or NaOH addition (103.12 ± 5.74%), compared to the other two protocols tested. TCA-acetone precipitates were difficult to solubilize, which negatively influenced recovery percentage (77.91 ± 8.79%) and band presence. M/C with ultrasonic homogenization showed an intermediate recovery between the other two protocols (94.22 ± 4.86%) without affecting protein pattern on SDS-PAGE. These precipitation methods affected the recovery of low MW proteins (< 15 kDa). CONCLUSIONS: These results help in the processing of samples of CHO cells for their proteomic study by means of an easily accessible, fast protocol, with an almost complete recovery of cellular proteins and the capture of the original complexity of the cellular composition. Acetone protocol could be incorporated to sample-preparation workflows in a straightforward manner and can probably be applied to other mammalian cell lines as well.