Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems

This work aimed to optimize the extraction of an extracellular protease produced by the cold-adapted yeast Rhodotorula mucilaginosa L7 using aqueous two-phase systems (ATPS) comprising polyethylene glycol (PEG) and sodium citrate or sodium tartrate. First, the biocompatibility of the phase forming agents was assessed. The results obtained with PEG-2000, PEG-4000, and PEG-6000 demonstrated that even at large PEG concentrations (32 wt%) the protease maintains its activity after 3 h of reaction, whereas an increase in salt concentration provokes a gradual decrease in protease stability. Subsequently, the partitioning of the protease in both types of ATPS was assessed, evaluating the effect of temperature, molecular weight, and concentration of PEG on protease purification, using two 2³-full factorial designs. The best partitioning conditions were obtained in PEG-6000/sodium tartrate-based ATPS, at 30ºC (with a yield of 81.09 ± 0.66% and a purification factor of 2.51 ± 0.03). Thus, considering the biodegradable characteristics of the system, the PEG/sodium tartrate ATPS is a viable and economic low-resolution step in protease purification, with a strong potential for future industrial application.     

Oligomeric poly(imide-amides)s containing side 1,8-naphthalimidyl groups: synthesis and characterization

Poly(imide-amide)s (PIAs) were synthesized from isophthalic acids, which have in position 5 a bulky group like 1,8-naphthalimidyl bonded to amino acids as flexible spacers, and the diamine bis(4-aminophenyl)diphenylsilane, which provides a polar group in the main chain. Glycine, l-alanine, l-phenylalanine, l-valine and p-aminobenzoic acid were used as amino acids. Polymers were obtained according to the Yamazaki method and characterized by elemental analysis, optical activity, IR and ¹H, ¹³C and ²⁹Si NMR spectroscopy. PIAs were soluble in aprotic polar solvents but not in common organic solvents, and obtained with low η _inh values, which was an indicative of low molecular weights species, probably of oligomeric nature. According to the polymeric structure, the only difference between PIAs is the structure of the amino acid residue, and in this sense it was possible to see an increase of the T _g values when the volume of the amino acid residue also increased, due to the lower possibility of internal mobility of the side chains. The higher Tg value was obtained with the PIA-e, which includes an aromatic ring as a side chain, derived from p-aminobenzoic acid. PIAs, in spite of the good TDT values obtained, were not thermally stable in the sense that the 10 % of weight lost was obtained at lower temperature than 400 °C, with the exception of PIA-e derived from p-aminobenzoic acid. However, there was an increase of the TDT values when the volume of the amino acid residue increases. The PIAs do not show good UV–vis transparence probably due to the low free volume of those including an aliphatic amino acid residue.     

Surface and interfacial fourier transform infrared spectroscopic studies of latexes. XIV. Surface phase separation in polystyrene/poly-n-butyl acrylate latex films

This study focuses on the behavior of sodium dioctylsulfosuccinate (SDOSS) in 50/50 w/w % polystyrene/poly(butyl acrylate) (p-Sty/p-BA) latex films. Specifically, mobility and orientation are examined in the context of the film formation by the use of dynamic mechanical thermal analysis and attenuated total reflectance (ATR) Fourier transform infrared (FT-IR) spectroscopy. While for the homopolymer blends of p-Sty and p-BA, two T_g values resulting from a phase separation of p-Sty and p-BA phases are observed, only a single T_g is detected for a copolymer of the same mixture, indicating a single phase within the film. ATR FTIR spectroscopic data indicate that the phase separation of p-Sty and p-BA blends does not occur uniformly across the film. After coalescence, p-Sty particles produce a significant degree of stratification at approximately 1.6 μm from the film surface. At this depth, the polystyrene rings assume preferentially parallel orientation to the film surface. At the same time, the hydrophilic groups of SDOSS surfactant (SO⁻₃Na⁺) are oriented preferentiallyparallel to the surface. Under high relative humidity conditions, water is able to diffuse into the film and swells the surface layers, thus causing them to expand. As a result, the top, predominately poly-n-BA surface becomes “thicker», and p-Sty phase appears to be near 2.3 μm from the surface. The polystyrene rings maintain their preferential parallel orientation to the surface, but the hydrophilic groups of SDOSS are able to diffuse into the film with the water uptake and are thus not present at the filmair interface. © 1996 John Wiley & Sons, Inc.     

OCTN1: A Widely Studied but Still Enigmatic Organic Cation Transporter Linked to Human Pathology and Drug Interactions

The Novel Organic Cation Transporter, OCTN1, is the first member of the OCTN subfamily; it belongs to the wider Solute Carrier family SLC22, which counts many members including cation and anion organic transporters. The tertiary structure has not been resolved for any cation organic transporter. The functional role of OCNT1 is still not well assessed despite the many functional studies so far conducted. The lack of a definitive identification of OCTN1 function can be attributed to the different experimental systems and methodologies adopted for studying each of the proposed ligands. Apart from the contradictory data, the international scientific community agrees on a role of OCTN1 in protecting cells and tissues from oxidative and/or inflammatory damage. Moreover, the involvement of this transporter in drug interactions and delivery has been well clarified, even though the exact profile of the transported/interacting molecules is still somehow confusing. Therefore, OCTN1 continues to be a hot topic in terms of its functional role and structure. This review focuses on the most recent advances on OCTN1 in terms of functional aspects, physiological roles, substrate specificity, drug interactions, tissue expression, and relationships with pathology.     

Strain-specific polyketide synthase genes of Aspergillus niger

In silico comparison of 34 putative pks genes in Aspergillus niger strain CBS 513.88 versus A. niger strain ATCC 1015 genome revealed significant nucleotide identity (>95% covering a minimum of 99% of the gene sequence) for 31 of these genes (approximately 91%). A. niger CBS 513.88 harbors three putative pks genes (An01g01130, An11g05940, and An15g07920), for which nucleotide identity was not found in A. niger ATCC 1015. To compare the results of the in silico analysis with the in vivo situation, experimental data were obtained for a large number of A. niger strains obtained from different substrates and geographical regions. Three putative pks genes that were found to be variable between the two A. niger strains using bioinformatics tools were in fact strain-specific genes based on experimental data. The PCR amplification signals for the An01g01130, An11g05940, and An15g07920 pks genes were detected in only 97%, 71%, and 26% of the strains, respectively. Southern blot analyses confirmed the PCR data. Because one of the strain-specific pks genes (An15g07920) is located in a putative ochratoxin cluster, we focused our investigation on that region. We assessed the ochratoxin production capability of the 119 A. niger strains and found a positive association between the presence of this pks gene and the capability of the respective strain to produce ochratoxin.     

Effect of ultraviolet processing on selected properties of egg white

The effect of ultraviolet processing (10.6 and 63.7 kJ m(-2)) on selected properties of egg white (absorbance, particle size, protein fractions, free sulfhydryl content, immunoreactivity, viscosity, gelling and foaming properties) was investigated. Ultraviolet exposure induced the development of browning, the formation of large protein aggregates by disulfide exchange, and protein backbone cleavage. However, egg white proteins were differently sensitive to UV radiation. No changes in immunoreactivity, gelling temperature and gel firmness were observed. Independently on the UV dose, light treated egg white produced foams with higher stability. This effect was attributed to protein aggregates jamming in the fluid interstices between bubbles and/or to the higher viscosity of the aqueous phase. The latter was also associated to higher foam volume.     

Nitrogen cycling in tropical grass-legume pastures managed under canopy light interception

Nutrient Cycling in Agroecosystems - In grass-legume pastures, grazing management strategies are an essential factor affecting nitrogen (N) cycling. This study assessed the impact of grazing...     

La CLAve to increase psychosis literacy of Spanish-speaking community residents and family caregivers.

The authors developed and tested a 35-min psychoeducational program with the goal of increasing Spanish-speaking persons’ literacy of psychosis. The program uses popular cultural icons derived from music, art, and videos, as well as a mnemonic device—La CLAve (The Clue)—to increase (a) knowledge of psychosis, (b) efficacy beliefs that one can identify psychosis in others, (c) attributions to mental illness, and (d) professional help-seeking. Assessments were conducted before and after administering the program to both community residents (n = 57) and family caregivers of persons with schizophrenia (n = 38). For community residents, the authors observed increases across the 4 domains of symptom knowledge, efficacy beliefs, illness attributions, and recommended help-seeking. For caregivers, increases were observed in symptom knowledge and efficacy beliefs. La CLAve is a conceptually informed psychoeducational tool with a developing empirical base aimed at helping Spanish-speaking Latinos with serious mental illness obtain care in a timely manner. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Von Willebrand disease: characteristics and response to desmopressin. Study of 103 cases

BACKGROUND: To describe the main characteristics and response to desmopressin infusion in 103 patients suffering from von Willebrand disease (vWD). PATIENTS AND METHODS: The criteria for diagnosis were (except for type 2N) the coexistence of von Willebrand factor ristocetin cofactor (vWF:RCo) activity < 50 U/dl with bleeding disease or one of the following data: von Willebrand factor antigen (vWF:Ag) activity < 50 U/dl, factor VIII (FVIII) activity < 50 U/dl or the existence of a increased bleeding time (BT). Multimeric studies of vWF were performed in 51 cases and ristocetin induced platelet aggregation (RIPA) was also performed. RESULTS: Spontaneous bleeding was found in 36 patients, while in 18 cases the diagnosis was done after surgical bleeding. Thirteen patients (6 presenting with mild bleeding) were studied for abnormalities in the routine preanestesic tests. Other 22 patients were diagnosed with vWD by familial studies. There were 3 patients with type 2B, 1 case with type 2N and other patient with type 3. BT was found increased in 26 out of 58 patients. The activities of vWF:CoR and vWF:Ag were 38.4 (9.4) U/dl and 45.8 (23.2) U/dl, respectively, while the activity of FVIII was 49.9 (20.8) U/dl. Prophylactic DDAVP (desmopressin) was infused in 32 patients. After 1 h, basal activities of vWF:CoR and vWF:Ag were increased by 3.1 (3.2) and 3.4 (3.1) times, respectively, and maintained for 3 h. FVIII activity increased 3.6 (2.3) times the basal levels decreasing after 3 h (2.9 [2.1]; p < 0.01). The BT was corrected in 8 out of ten patients. CONCLUSIONS: vWD is a major cause of surgical bleeding. Preanestesic anamnesis and coagulation tests can be useful to identify vWD. Many patients with vWD have normal BT. A failure in the response to desmopressin infusion is unusual.