Observance of polymorphic behaviour during dissolution of insulin and lysozyme

Although protein crystallization is a unit operation with potentially high separation factors, it has not been widely used in industry. Protein crystallization studies and practices have hitherto been largely limited to crystallography protocols. Knowledge of the behaviour of protein in solution would help to overcome empiric limitations in protein crystallisation. Thus, dissolution of porcine insulin and hen egg white lysozyme was studied and an unusual variation in solute concentration, with a concentration peak for short dissolution times, was verified. Polymorphic behaviour of protein in solution was observed, which altered physical properties such as solubility.22333133

Aqueous extraction of recombinant human proinsulin from transgenic maize endosperm

Different plant species have been used as systems to produce recombinant proteins. Maize is a crop considered to have a large potential to produce high levels of recombinant proteins and is the host for the recombinant proteins from plants currently available on the market. In the development of a plant system to produce a recombinant proteins it is important to consider the costs related to downstream processing. Also, the steps necessary to achieve the protein purity required will be highly influenced by the quality of the extract obtained. In this study, we analyzed aqueous extracts from the endosperm of transgenic maize expressing recombinant human proinsulin (rhProinsulin). A study of the effects of the variables pH and ionic strength on the extraction efficiency was carried out using experimental design and response surface methodology. Besides the concentration of the recombinant protein, the characteristics of the extracts were evaluated in terms of concentration of native components (proteins, carbohydrates, and phenolic compounds) and extract filterability. The highest rhProinsulin concentration (97.33 ng/mL) was found with a 200 mM NaCl pH 10.0 extraction solution. Under this experimental condition the concentrations of total soluble proteins, carbohydrates, and phenolics were 2.01 mg/mL, 2.21 mg/mL, and 0.11 mmol/L, respectively.2151466147

Application of Electrochemically Produced Aluminum Hydroxide Gel for Prepurification of Recombinant Synthetic Green Fluorescent Protein Produced in Tobacco Leaves

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)The use of recombinant proteins has increased greatly in recent years, as also have increased the number of techniques and materials used for their production and purification. Among the different types of bioreactors being studied, there is a general consensus among scientists that production in green plant tissues such as leaves is more feasible. However, the presence of chlorophyll and phenolic compounds in plant extracts, which can precipitate and denature the proteins besides damaging separation membranes and gels, makes this technology impracticable on a commercial scale. In the present work, the adsorption to electrochemically produced aluminum hydroxide gel was applied as a prepurification step for recombinant synthetic green fluorescent protein (sGFP), also referred to as enhanced green fluorescent protein, produced in Nicotiana benthamiana leaves. Removal efficiencies of 99.7% of chlorophyll, 88.5% of phenolic compounds, and 38.5% of native proteins from the N. benthamiana extracts were achieved without removing sGFP from the extracts. As electrochemical preparation of aluminum hydroxide gel is a cost-effective technique, its use can substantially contribute to the development of future production platforms for recombinant proteins produced in green plant tissues of pharmaceutical and industrial interest. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 27: 1029-1035, 201127410291035Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

TERNARY PHASE DIAGRAM OF KETAMINE ((R,S)-2-(2-CHLOROPHENYL)-2-METHYLAMINOCYCLOHEXANONE) IN ETHANOL AND PRELIMINARY STUDIES AIMING AT ENANTIOSELECTIVE CRYSTALLIZATION OF S-KETAMINE

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Crystallization is an important industrial-scale process for the purification of enantiomers that depends on a phase diagram. In this work, the ternary phase diagram of R- and S-ketamine in ethanol was determined. The eutectic point indicated that crystallization of pure enantiomers from solutions containing more than 75% of the desired enantiomer is feasible. Solubility studies showed the feasibility of using temperature control to conduct the process. Batch crystallization of ketamine (S/R:80/20) solutions at 25 degrees C provided the isolation of S-ketamine (purity of 100%) with a yield from 65 to 70% and a productivity of 6.5 g/(1h).262427434Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)UnicampFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

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Abstract:  Postpartum women experience cognitive modifications evidenced in the resulting production of verbal fluency tests (VFT) (Miranda, 2017). Verbal fluency contains associative and cognitive flexibility components (Horcajuelo, 2014). There are traditional statistical techniques for its study, but Exploratory Factor Analysis (EFA) is proposed because it allows the interpretation of extensive measures, provides a more parsimonious representation and is of simple applicability (Fabrigar, 2011; Martinez, 2012). The objective was to optimize the study of the verbal fluency of puerperal women in the province of Córdoba (Argentina) during the period 2016-2020 through EFA. It was an observational, cross-sectional, descriptive and correlational study. An accidental non-probability sampling was carried out with 100 lactating women as a sample. Demographic information was collected and four valid versions of VFT were used (semantic -Animals- and phonological -letter p, f and excluded A) (Fumagalli, 2018). An analysis of tetrachoric correlations was carried out and classical criteria (loads >0.30, eigenvalue, variance and communality) were considered (Mavrou, 2015; Miranda, 2019). Rotation methods and factor structures were compared according to demographic characteristics. Using the programs InfoStat and Stata 14. The EFA showed similar results to the traditional methods, but the mathematical clarity in the EFA allows us to estimate similarity, uniqueness and commonality (Barbero García, 2011). The promax rotation yielded more robust results. After educational level, parity was the variable that had the most effects on VFT. Parity was associated with intrusions (β = 0.37), isolated words (β = 0.28) and jumps (β = 0.33) in the VFT-p; number of clusters (β = 0.30) in VFT-excluded A; and with jumps in VFT-f (β = 0.27) and number of groups for animals (β = 0.22). EFA is a technique with potential for the study of word clustering. The estimation of the semantic structure and the grouping strategies allow understanding cognitive processes, in addition to providing the theoretical bases to analyze these processes under pathological conditions (Akiyama, 2018; Shao, 2014). The study of cognitive variations in the puerperium has received more interest lately. This shows the holistic approach of the puerperal woman, understanding other health processes in this vital stage.Resumen:  Las mujeres puérperas experimentan modificaciones cognitivas evidenciadas en la producción resultante de pruebas de fluidez verbal (PFV) (Miranda, 2017). La fluidez verbal contiene componentes asociativos y de flexibilidad cognitiva (Horcajuelo, 2014). Existen técnicas estadísticas tradicionales para su estudio,  pero se propone el Análisis Factorial Exploratorio (AFE) porque permite interpretar medidas extensas, provee una representación más parsimoniosa y es de simple aplicabilidad (Fabrigar, 2011; Martinez, 2012). El objetivo fue optimizar el estudio de la fluidez verbal de mujeres puérperas de la provincia de Córdoba (Argentina) durante el período 2016-2020 mediante AFE. Fue un estudio observacional, transversal, descriptivo y correlacional. Se realizó un muestreo no probabilístico accidental con 100 mujeres lactantes como muestra. Se recolectó información demográfica y se utilizaron cuatro versiones válidas de PFV (semántica -Animales- y fonológicas -letra p, f y excluida A) (Fumagalli, 2018). Se ejecutó un análisis de correlaciones tetracóricas y se consideraron criterios clásicos (cargas >0,30, valor propio, varianza y comunalidad) (Mavrou, 2015; Miranda, 2019). Se compararon métodos de rotación y estructuras factoriales según características demográficas. Utilizando los programas InfoStat y Stata 14. El AFE mostró resultados similares a los métodos tradicionales, pero la claridad matemática en el AFE permite estimar similaridad, unicidad y comunalidad (Barbero García, 2011). La rotación promax arrojó resultados más robustos. Después del nivel educativo, la paridad fue la variable que tuvo más efectos sobre las PFV. La paridad se asoció con intrusiones (β=0,37), palabras aisladas (β=0,28) y saltos (β=0,33) en la PFV-p; número de agrupaciones (β=0,30) en PFV-excluida A; y con saltos en PFV-f (β=0,27) y número de agrupaciones para animales (β=0,22). El AFE es una técnica con potencial para el estudio del agrupamiento de palabras. La estimación de la estructura semántica y las estrategias de agrupamiento permiten comprender procesos cognitivos, además de aportar las bases teóricas para analizar dichos procesos en condiciones patológicas (Akiyama, 2018; Shao, 2014). El estudio de las variaciones cognitivas en el puerperio ha cobrado mayor interés últimamente. Esto evidencia el abordaje holístico de la mujer puérpera, comprendiendo otros procesos de salud en esta etapa vital.

Associations between reliable changes in depression and changes in BMI, total body fatness and visceral adiposity during a 12-month weight loss trial.

We investigated associations between changes in depression and body composition over a 12-month weight loss trial. Of the 298 adults (BMI > 27 m/kg2), 219 with complete depression and body composition data were included. A 10-item Center for Epidemiologic Studies Depression Scale measured depression; dual-energy X-ray absorptiometry measured body composition. Multinomial logistic regression predicted reliable changes in depression by BMI, body fat (BF) and visceral adiposity (VAT). Multiplicative interaction terms tested modification by sex and ethnicity. Participants with increases in body composition were less likely to experience improvements in depression (BMI: RRR = 0.79 (0.68-0.91), p < 0.01; BF: RRR = 0.97 (0.94 - 0.99), p = 0.01; VAT: RRR = 0.99 (0.98-1.00), p = 0.02), but not worsening of depression (BMI: RRR = 1.29 (0.96-1.73), p = 0.10; BF: RRR = 1.04 (0.99-1.09), p = 0.15; VAT: RRR = 1.01 (1.00-1.03), p = 0.18). Sex and ethnicity interaction terms were not significant. However, the relationship was only significant among females, among non-Latinos for BMI and BF, and among Latinos for VAT. Our study supports the association between depression and obesity and highlights the need for longitudinal studies investigating VAT and depression in diverse ethnic groups

ON THE USE OF THE PHOTOACOUSTIC EFFECT FOR INVESTIGATING PHASE-TRANSITIONS IN SOLIDS

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Homogeneous Bispecifics by Disulfide Bridging

We report on a chemical platform to generate site-specific, homogeneous, antibody-antibody conjugates by targeting and bridging disulfide bonds. A bispecific antibody construct was produced in good yield through simple reduction and bridging of antibody fragment disulfide bonds, using a readily synthesized bis-dibromomaleimide cross-linker. Binding activity of antibodies was maintained, and in vitro binding of target antigens was observed. This technology is demonstrated through linking scFv and Fab antibody fragments, showing its potential for the construction of a diverse range of bispecifics

Combining ability of summer-squash lines with different degrees of parthenocarpy and PRSV-W resistance

The aim was to assess heterosis in a set of 16 summer-squash hybrids, and evaluate the combining capacity of the respective parental lines, which differed as to the degree of parthenocarpy and resistance to PRSV-W (Papaya Ringspot Virus-Watermelon strain). The hybrids were obtained using a partial diallel cross design (4 × 4). The lines of parental group I were 1 = ABX-037G-77-03-05-01-01-bulk, 2 = ABX-037G-77-03-05-03-10-bulk, 3 = ABX-037G-77-03-05-01-04-bulk and 4 = ABX-037G-77-03-05-05-01-bulk, and of group II, 1′ = ABX-037G-77-03-05-04-08-bulk, 2′ = ABX-037G-77-03-05-02-11-bulk, 3′ = Clarice and 4′ = Caserta. The 16 hybrids and eight parental lines were evaluated for PRSV-W resistance, parthenocarpic expression and yield in randomized complete-block designs, with three replications. Parthenocarpy and the resistance to PRSV-W were rated by means of a scale from 1 to 5, where 1 = non-parthenocarpic or high resistance to PRSV-W, and 5 = parthenocarpic or high susceptibility to PRSV-W. Both additive and non-additive gene effects were important in the expression of parthenocarpy and resistance to PRSV-W. Whereas estimates of heterosis in parthenocarpy usually tended towards a higher degree, resistance to PRSV-W was towards higher susceptibility. At least one F1 hybrid was identified with a satisfactory degree of parthenocarpy, resistance to PRSV-W and high fruit-yield