Immobilization of Horseradish Peroxidase onto Montmorillonite/Glucosamine-Chitosan Composite for Electrochemical Biosensing of Polyphenols.

Glucosamine-chitosan synthesized by the Maillard reaction was combined with montmorillonite to obtain a nanohybrid composite to immobilize horseradish peroxidase. The material combines the advantageous properties of clay with those of the chitosan derivative; has improved water solubility and reduced molecular weight and viscosity; involves an eco-friendly synthesis; and exhibits ion exchange capacity, good adhesiveness, and a large specific surface area for enzyme adsorption. The physicochemical characteristics of the composite were analyzed by infrared spectroscopy and X-ray diffraction to determine clay-polycation interactions. The electrochemical response of the different polyphenols to glassy carbon electrodes modified with the composite was evaluated by cyclic voltammetry. The sensitivity and detection limit values obtained with the biosensor toward hydroquinone, chlorogenic acid, catechol, and resorcinol are (1.6 ± 0.2) × 102 µA mM-1 and (74 ± 8) nM; (1.2 ± 0.1) × 102 µA mM-1 and (26 ± 3) nM; (16 ± 2) µA mM-1 and (0.74 ± 0.09) µM; and (3.7± 0.3) µA mM-1 and (3.3 ± 0.2) µM, respectively. The biosensor was applied to quantify polyphenols in pennyroyal and lemon verbena extracts.

Hematology and serum biochemistry of broilers fed diets supplemented with chondroitin and glucosamine sulfates

The objective of this study was to evaluate the hematology and serum biochemistry of broilers fed diets supplemented with chondroitin and glucosamine sulfates. An experiment was laid out in a completely randomized design with a 3 × 3 factorial arrangement (three levels of chondroitin sulfate: 0, 0.05 and 0.10%; and three levels of glucosamine sulfate: 0, 0.15, and 0.30%), with each treatment involving six replicates of 30 birds. Hematology (red blood cells, hemoglobin, hematocrit, total plasma protein [TPP], thrombocytes, white blood cells, eosinophils, monocytes, heterophils, and lymphocytes) and serum biochemistry (totalinteraction between sulfates influenced (p < 0.05) total calcium by 21 days, ionic calcium by 21 and 42 days, and phosphorus, chlorides, and sodium by 42 days. Supplementation with chondroitin and glucosamine sulfates in the broilers' diet favored their immune system and mineral metabolism, increasing serum concentrations of calcium, phosphorus, and sodium. serum protein [TSP], albumin, globulins, aspartate aminotransferase [AST], gamma-glutamyltransferase [GGT], alkaline phosphatase [AP], total calcium, ionic calcium, phosphorus, sodium, potassium, and chlorides) variables were evaluated at 21 and 42 days. Data were subjected to analysis of variance. When the means differed significantly by the F-test, orthogonal analysis was performed to test the linear and quadratic effects of chondroitin and glucosamine sulfate levels. Glucosamine sulfate reduced the number of monocytes linearly, by 42 days (p = 0.0399). There was an interaction effect between the sulfates on total white blood cells (p = 0.0099) and lymphocytes (p = 0.0004) by 21 days. Chickens supplemented with 0.10% chondroitin sulfate showed a linear increase in white blood cells (p = 0.0287) and lymphocytes (p = 0.0144) with the addition of glucosamine sulfate. Chondroitin sulfate supplementation increased serum albumin linearly (p = 0.0099) and TSP quadratically (p = 0.0140), by 21 days. Glucosamine sulfate induced a quadratic response (p < 0.05) in albumin by 42 days, with the lowest value found with the inclusion of 0.06%. Glucosamine sulfate reduced chlorides linearly (p = 0.0237) by 21 days and increased calcium linearly (p = 0.0012) by 42 days. The interaction between sulfates influenced (p < 0.05) total calcium by 21 days, ionic calcium by 21 and 42 days, and phosphorus, chlorides, and sodium by 42 days. Supplementation with chondroitin and glucosamine sulfates in the broilers' diet favored their immune system and mineral metabolism, increasing serum concentrations of calcium, phosphorus, and sodium.(AU)

Objetivou-se avaliar a hematologia e a bioquímica sérica de frangos de corte suplementados com sulfatos de condroitina e de glucosamina na ração. Foi conduzido um experimento em delineamento inteiramente casualizado, em esquema fatorial 3 x 3 (três níveis de sulfato de condroitina: 0; 0,05 e 0,10%; e três níveis de sulfato de glucosamina: 0; 0,15 e 0,30%), cada tratamento com seis repetições de 30 aves. Foram avaliadas as variáveis de hematologia (hemácias, hemoglobina, hematócrito, proteínas plasmáticas totais [PPT], trombócitos, leucócitos, eosinófilos, monócitos, heterofilos e linfócitos) e bioquímica sérica (proteínas séricas totais [PST], albumina, globulinas, aspartato aminotransferase [AST], gama glutamiltransferase [GGT], fosfatase alcalina [FA], cálcio total, cálcio iônico, fósforo, sódio, potássio e cloretos) aos 21 e 42 dias. Os dados foram submetidos à análise de variância. Quando as médias diferiram significativamente pelo teste F, a análise ortogonal foi realizada para testar os efeitos lineares e quadráticos dos níveis dos sulfatos de condroitina e de glucosamina. Observou-se efeito linear decrescente (p = 0,0399) do sulfato de glucosamina na quantidade de monócitos aos 42 dias. Houve interação dos sulfatos para leucócitos totais (p = 0,0099) e linfócitos (p = 0,0004) aos 21 dias. Frangos suplementados com 0,10% de sulfato de condroitina mostraram um aumento linear dos leucócitos (p = 0,0287) e dos linfócitos (p = 0,0144) com a inclusão de sulfato de glucosamina. A suplementação com sulfato de condroitina aumentou linearmente (p = 0,0099) a albumina sérica e afetou de forma quadrática (p = 0,0140) as PST aos 21 dias. O sulfato de glucosamina demonstrou um efeito quadrático (p < 0,05) sobre a albumina aos 42 dias, o menor valor foi encontrado para a inclusão de 0,06%, respectivamente. O sulfato de glucosamina reduziu linearmente (p = 0,0237) os cloretos aos 21 dias e aumentou linearmente (p = 0,0012) o cálcio total aos 42 dias. Verificouse interação (p < 0,05) dos sulfatos para cálcio total aos 21 dias, cálcio iônico aos 21 e 42 dias e para fósforo, cloretos e sódio aos 42 dias. A suplementação com os sulfatos de condroitina e de glucosamina na ração de frangos de corte favoreceram o sistema imune e o metabolismo de minerais, com aumento nas concentrações séricas de cálcio, fósforo e sódio.(AU)

The effect of deproteinization methods on the properties of glucosamine hydrochloride from shells of white leg shrimp (Litopenaeus vannamei) and black tiger shrimp (Penaeus monodon) / Efeito de métodos de desproteinização nas propriedades do cloridrato de glucosamina de cascas de camarão de pernas brancas (Litopenaeus vannamei) e cascas de camarão de tigre preto (Penaeus monodon)

The effect of methods to remove protein content on the properties of glucosamine hydrochloride from the shells of white leg shrimp (Litopenaeus vannamei) and black tiger shrimp (Penaeus monodon) was investigated. Chitin from shrimp shells was obtained by demineralization in 6% HCl for 12h, deproteinization by two different methods (first group soaked in 8% NaOH for 36h and second group treated in Alcalase enzyme at the concentration of 0.2% for 36h). Two group samples were converted to glucosamine hydrochloride by soaking in 36.76% HCl solution for 5h at 85 °C. The results of fourier transform infrared spectroscopy (FTIR), solubility and recovery yield analysis showed that deproteinization methods did not significantly affect the properties of glucosamine hydrochloride. However, glucosamine hydrochloride from white leg shrimp shells contained higher recovery yield and solubility than black tiger shrimp shells.(AU)

Investigou-se o efeito de métodos para remover o conteúdo de proteínas nas propriedades do cloridrato de glucosamina das conchas de camarão de pernas brancas (Litopenaeus vannamei) e camarão de tigre preto (Penaeus monodon). A quitina das cascas de camarão foi obtida por desmineralização em HCl a 6% por 12 h, desproteinização por dois métodos diferentes (primeiro grupo embebido em NaOH a 8% por 36 h e segundo grupo tratado na enzima Alcalase na concentração de 0,2% por 36 h). Duas amostras de grupo foram convertidas em cloridrato de glucosamina por imersão em solução de 12M HCl por 5 h a 85 °C. Os resultados das análises de FTIR, solubilidade e rendimento de recuperação mostraram que os métodos de desproteinização não afetaram significativamente as propriedades do cloridrato de glucosamina. No entanto, o cloridrato de glucosamina de cascas de camarão de pernas brancas continha maior rendimento e solubilidade de recuperação do que as cascas de camarão tigre preto.(AU)

The effect of deproteinization methods on the properties of glucosamine hydrochloride from shells of white leg shrimp (Litopenaeus vannamei) and black tiger shrimp (Penaeus monodon) / Efeito de métodos de desproteinização nas propriedades do cloridrato de glucosamina de cascas de camarão de pernas brancas (Litopenaeus vannamei) e cascas de camarão de tigre preto (Penaeus monodon)

The effect of methods to remove protein content on the properties of glucosamine hydrochloride from the shells of white leg shrimp (Litopenaeus vannamei) and black tiger shrimp (Penaeus monodon) was investigated. Chitin from shrimp shells was obtained by demineralization in 6% HCl for 12h, deproteinization by two different methods (first group soaked in 8% NaOH for 36h and second group treated in Alcalase enzyme at the concentration of 0.2% for 36h). Two group samples were converted to glucosamine hydrochloride by soaking in 36.76% HCl solution for 5h at 85 °C. The results of fourier transform infrared spectroscopy (FTIR), solubility and recovery yield analysis showed that deproteinization methods did not significantly affect the properties of glucosamine hydrochloride. However, glucosamine hydrochloride from white leg shrimp shells contained higher recovery yield and solubility than black tiger shrimp shells.

Investigou-se o efeito de métodos para remover o conteúdo de proteínas nas propriedades do cloridrato de glucosamina das conchas de camarão de pernas brancas (Litopenaeus vannamei) e camarão de tigre preto (Penaeus monodon). A quitina das cascas de camarão foi obtida por desmineralização em HCl a 6% por 12 h, desproteinização por dois métodos diferentes (primeiro grupo embebido em NaOH a 8% por 36 h e segundo grupo tratado na enzima Alcalase na concentração de 0,2% por 36 h). Duas amostras de grupo foram convertidas em cloridrato de glucosamina por imersão em solução de 12M HCl por 5 h a 85 °C. Os resultados das análises de FTIR, solubilidade e rendimento de recuperação mostraram que os métodos de desproteinização não afetaram significativamente as propriedades do cloridrato de glucosamina. No entanto, o cloridrato de glucosamina de cascas de camarão de pernas brancas continha maior rendimento e solubilidade de recuperação do que as cascas de camarão tigre preto.

Systematic Review and Study of S Curves for Biomass Quantification in Solid-state Fermentation (SSF) and Digital Image Processing (DIP) Applied to Biomass Measurement in Food Processes.

BACKGROUND: There are several methods for the quantification of biomass in SSF, such as glucosamine measurement, ergosterol content, protein concentration, change in dry weight or evolution of CO2 production. However, all have drawbacks when obtaining accurate data on the progress of the SSF due to the dispersion in cell growth on the solid substrate, and the difficulty encountered in separating the biomass. Studying the disadvantages associated with the process of biomass quantification in SSF, the monitoring of the growth of biomass by a technique known as digital image processing (DIP), consists of obtaining information on the production of different compounds during fermentation, using colorimetric methods based on the pixels that are obtained from photographs. OBJECTIVE: The purpose of this study was to know about the state of the technology and the advantages of DIP. METHODS: The methodology employed four phases; the first describes the search equations for the SSF and the DIP. A search for patents related to SSF and DIP carried out in the Free Patents Online and Patent inspiration databases. Then there is the selection of the most relevant articles in each of the technologies. As a third step, modifications for obtaining the best adjustments were also carried out. Finally, the analysis of the results was done and the inflection years were determined by means of six mathematical models widely studied. RESULTS: For these models, the inflection years were 2018 and 2019 for both the SSF and the DIP. Additionally, the main methods for the measurement of biomass in SSF were found, and are also indicated in the review, as DIP measurement processes have already been carried out using the same technology. CONCLUSION: In addition, the DIP has shown satisfactory results and could be an interesting alternative for biomass measurement in SSF, due to its ease and versatility.

Sustainable production with obtaining glucosamine from crab exoskeletons / Produção sustentável para obtenção de glucosamina a partir de exoesqueletos de caranguejo

Two chemical treatments, five enzymatic (pectinase, lipase, hemicellulase, hemicellulose-cellulase or lipase-pectinase) and one microbiological (Bacillus subtilis) treatment were evaluated to obtain glucosamine hydrochloride (Gluc-HCl) from the chitin obtained from crab (Callinectes bellicosus) exoskeletons. Chemical treatments were referred as Method A (HCl hydrolysis during 75 min at 90°C) and Method B (HCl hydrolysis during 20 min and 14 h of rest). Glucosamine and, in some cases, N-acetyl-D-glucosamine were identified and quantified by HPLC. Treatments with the greater concentrations of Gluc-HCl in descending order were: lipase (94.4 mg/g), microbiological (45.7 mg/g), lipase-pectinase (22.9 mg/g), hemicellulase-cellulase (20.9 mg/g), hemicellulase (15.3 mg/g), pectinase (10.7 mg/g), Chemical A (7.3mg/g) and Chemical B (7.3mg/g). In terms of yield, the best treatments in descending order were: pectinase (94%), microbiological (94%), hemicellulase (92%), lipase (91%), Chemical B (88%), lipase-pectinase (88%), hemicellulase-cellulase (86%) and Chemical A (28.5%). The two most profound treatments were lipase and microbiological, so they are proposed as part of a viable method to produce Gluc-HCl from crab exoskeletons; they are ecofriendly procedures and could add value to the crab's productive chain.(AU)

Dois tratamentos químicos, cinco enzimáticos (pectinase, lipase, hemicelulase, hemicelulose-celulase ou lipase-pectinase) e um microbiológico (Bacillus subtilis) foram avaliados para obter o cloridrato de glucosamina (Gluc-HCl) da quitina obtida a partir de exoesqueletos de caranguejo (Callinectes. Bellicosus). Os dois tratamentos químicos foram nomeados como método A (hidrólise de HCl para 75 min a 90 °C) e método B (hidrólise de HCl para 20 min e 14 h de repouso). A Glucosamina e, em alguns casos, N-acetil-D-glucosamina foram identificados e quantificados por HPLC. Os tratamentos em que as melhores concentrações de Glucosamina-HCl foram obtidas, em ordem decrescente: lipase (94,4 mg/g), microbiológica (45,7 mg/g), lipase-pectinase (22,9 mg/g), hemicelulase-celulase (20,9 mg/g), hemicelulase (15,3 mg/g), pectinase (10,7mg/g), Quïmica A (7,3 mg/g) e Quïmica B (7,3 mg/g). Em termos de produtividade, os melhores tratamentos em ordem decrescente foram: pectinase (94%), microbiológica (94%), hemicelulase (92%), lipase (91%), química B (88%), lipase-pectinase (88%), hemicelulase- celulase (86%) e produto químico A (28,5%). Os dois melhores tratamentos foram lipase e microbiológicos, propostos como método viável para obtenção de Gluc-HCl a partir de exoesqueletos de caranguejo; cumprem procedimentos ecologicamente corretos e podem agregar valor à cadeia produtiva do caranguejo.(AU)

Caffeine and glucosamine mobility shifts by adduction with 2-butanol depended on interaction energy, charge delocalization, and steric hindrance in ion mobility spectrometry.

Ion mobility spectrometry (IMS) is an analytical technique that separates gas-phase ions drifting under an electric field according to their size to charge ratio. We used electrospray ionization-drift tube IMS coupled to quadrupole mass spectrometry to measure the mobilities of glucosamine (GH+ ) and caffeine (CH+ ) ions in pure nitrogen or when the shift reagent (SR) 2-butanol was introduced in the drift gas at 6.9 mmol m-3 . Binding energies of 2-butanol-ion adducts were calculated using Gaussian 09 at the CAMB3LYP/6-311++G(d,p) level of theory. The mobility shifts with the introduction of 2-butanol in the drift gas were -2.4% (GH+ ) and -1.7% (CH+ ) and were due to clustering of GH+ and CH+ with 2-butanol. The formation of GBH+ was favored over that of CBH+ because GBH+ formed more stable hydrogen bonds (83.3 kJ/mol) than CBH+ (81.7 kJ/mol) for the reason that the positive charge on CH+ is less sterically available than on GH+ and the charge is stabilized by resonance in CH+ . These results are a confirmation of the arguments used to explain the drift behavior of these ions when ethyl lactate SR was used (Bull Kor Chem Soc 2014, 1023-1028). This study is a step forward to predict IMS separations of overlapping peaks in IMS spectra, simplifying a procedure that is trial and error by now.

Análise das propriedades físico-químicas da glucosamina sulfato comercial, uma matéria- prima utilizada em formulações farmacêuticas / Physicochemical properties analysis of commercial glucosamine sulfate, a raw material used in pharmaceutical formulations

Introdução: A qualidade dos produtos em farmácias de manipulação é determinada pela Agência Nacional de Vigilância Sanitária (ANVISA), mas os métodos descritos podem não ser adequados para analisar seus aspectos físico-químicos. Objetivo: Comparar aspectos físico-químicos da glucosamina sulfato de dois diferentes fornecedores com análises realizadas na farmácia de manipulação. Métodos: As características organolépticas, pH, solubilidade e densidade da glucosamina (n=50) dos fornecedores foram analisadas conforme descrito na Farmacopeia Brasileira e Compêndio Oficial e comparados aos laudos técnicos dos produtos adquiridos. Usaram-se os testes de Kolmogorov-Smirnov, coeficiente de correlação intraclasse (CCI) e Bland-Altman. Resultados: A análise de CCIevidenciou baixa reprodutibilidade para o teste de pH e densidade, e a análise de Bland-Altman demonstrou que os fornecedores subestimavam ou superestimavam os valores de pH e densidade em relação à farmácia. Conclusão: Os aspectos físico-químicos estão adequados conforme orientações da Anvisa, mas novas técnicas mais sensíveis devem ser utilizadas para garantir a qualidade da glucosamina nas formulações.

Introduction: The raw materials quality in the manipulation pharmacies are determined by Brazilian Health Surveillance Agency (ANVISA), but the protocols described may not be appropriate to analyze their physicochemical properties. Objective: To compare the physicochemical properties of glucosamine sulfate from two different suppliers with results obtained in the manipulation pharmacy. Methods: The organoleptic characteristics, pH, solubility and density of glucosamine samples (n=50) were analyzed according to the Brazilian Pharmacopoeia and Official Compendium and compared the technical reports of the suppliers. The results were analyzed by the Kolmogorov-Smirnov test, intraclass correlation coefficient (CCI) and Bland-Altman. Results: CCI analyses showed low reproducibility for pH and density test in the samples tested. In addition, Bland-Altman analysis indicated pH values and density of suppliers were underestimated or overestimated compared to the pharmacy. Conclusion: Physicochemical properties of glucosamine are appropriate according to Anvisa specifications, but new more sensitive techniques should be employed to ensure the glucosamine quality in the formulations.

Chondroitin sulfate and glucosamine in the cartilage and subchondral bone repair of dogs - Histological findings / Sulfato de condroitina e glucosamina na reparação da cartilagem e do osso subcondral de cães - Achados histológicos

Chondroitin and glucosamine sulfate nutraceuticals are commonly used in the management of degenerative articular disease in veterinary routine. However, there are controversies on the contribution of these substances to articular cartilage. The purpose of this study was to evaluate the efficiency of a chondroitin and glucosamine sulfate-based veterinary nutraceutical on the repair of an induced osteochondral defect in a dog femoral condyle, by macroscopic, histological and histomorphometric analyses. The nutraceutical was orally administered the day following injury induction, every 24 hours (treated group, TG, n=24), compared with animals that did not receive the product (control group, CG, n=24). Six animals per group were anaesthetized for sample collection at 15, 30, 60 and 90 days after surgery. At 15 days, defects were macroscopically filled with red-pinkish tissue. After 30 days, whitish color tissue was observed, both in TG and CG animals, with firmer consistency to touch at 60 and 90 postoperative days. Histological analysis demonstrated that, in both groups, there was initial blood clot formation, which was subsequently substituted by a fibrin net, with capillary proliferation from the adjacent bone marrow and infiltration of mesenchymal cells in clot periphery. As cellular differentiation developed, repair tissue presented a fibrocartilage aspect most of the time, and new subchondral bone formation occurred in the deepest area corresponding to the defect. Histomorphometry suggested that the nutraceutical did not favor the articular cartilage repair process. It was concluded that nutraceutical did not significantly influence chondrocytes proliferation or hyaline architecture restoration.(AU)

Os nutracêuticos compostos de sulfato de condroitina e glucosamina são comumente utilizados no manejo da doença articular degenerativa na rotina veterinária. Entretanto, existem controvérsias sobre a contribuição dessas substâncias à cartilagem articular. O objetivo deste trabalho foi avaliar a eficácia de um nutracêutico veterinário à base de sulfato de condroitina e glucosamina na reparação de defeitos osteocondrais induzidos no côndilo femoral de cães, através de análises macroscópica, histológica e histomorfométrica. O nutracêutico foi administrado no dia seguinte à indução da lesão, pela via oral, a cada 24 horas (grupo tratado - GT, 24 animais), sendo comparado a animais que não receberam o produto (grupo controle - GC, de igual número de animais). Aos 15, 30, 60 e 90 dias após a cirurgia, seis animais por grupo foram anestesiados para ser realizada a coleta das amostras. Aos 15 dias, os defeitos eram macroscopicamente preenchidos por tecido de coloração rósea a avermelhada. Já a partir dos 30 dias, observou-se preenchimento por tecido de coloração esbranquiçada, tanto nos animais do GT quanto nos do GC, com consistência mais firme ao toque digital aos 60 e 90 dias de pós-operatório. A análise histológica revelou que, em ambos os grupos, houve inicialmente formação de coágulo sanguíneo que, posteriormente, foi substituído por uma rede de fibrina, com proliferação de capilares a partir da medula óssea adjacente e infiltração de células mesenquimais na periferia do coágulo. À medida que se processou a diferenciação celular, o tecido de reparação se apresentou na maioria das vezes com aspecto de fibrocartilagem e, na região mais profunda da área correspondente ao defeito, ocorreu formação de osso novo subcondral. A histomorfometria sugeriu que o nutracêutico não favoreceu o processo de reparação da cartilagem articular. Concluiu-se que o nutracêutico não influenciou consideravelmente na proliferação de condrócitos nem na restauração da arquitetura hialina.(AU)

Effect of particle size and ammonium sulfate concentration on rice bran fermentation with the fungus Rhizopus oryzae.

The effects of rice bran particle size (0.18-0.39mm) and ammonium sulfate concentration in the nutrient solution (2-8g/L) on biomass production, protein and phenolic content generated by solid state fermentation with the fungus Rhizopus oryzae (CCT 1217) were studied. Particle size had a positive effect on biomass production and a negative effect (p⩽0.05) on protein and phenolic contents. Ammonium sulfate concentration had a positive effect (p⩽0.05) on biomass and phenolic content gain. Cultivation of fungus in rice bran with particle size of 0.18mm and in the presence of 8g/L ammonium sulfate, resulted in protein levels of 20g/100g dry wt and phenolics content of 4mg/g dry wt. These values were 53 and 65% higher than those achieved with unfermented rice bran. The results demonstrate that the fermentation process increased the value of compounds recovered for potential use in food formulations.

A matrix-assisted laser desorption/ionization mass spectrometry approach to the lipid A from Mesorhizobium loti.

The isolation, purification and analysis of the lipid A obtained from Mesorhizobium loti Ayac 1 BII strain is presented. Analysis of the carbohydrate moiety after acid hydrolysis by high-pH anion-exchange chromatography with pulse amperometric detection (HPAEC-PAD) showed the presence of glucosamine and galacturonic acid as the only sugar components. Gas chromatographic (GC) and GC/mass spectrometric (MS) analysis of the fatty acids revealed the presence of 3-OH-C12:0; 3-OH-C13:0; 3-OH-C20:0 and 27-OH-C28:0 among the major hydroxylated species. In addition, C16:0, C17:0, C18:0 and C 20:0 were shown as main saturated fatty acids. Different polyacylated species were evidenced by thin layer chromatography of lipid A, allowing the purification of two fractions. Ultraviolet matrix-assisted laser desorption/ionization time-of-flight (UV-MALDI-TOF) MS analysis with different matrices, in the positive- and negative-ion mode, was performed. The fast moving component revealed the presence of hexa-acylated species, varying in the fatty acid composition. Species containing three 3-OH fatty acids and a 27-OH-C28:0 fatty acid were observed. Individual ions within this family differ by +/-14 mass units. The slow moving component was enriched mainly in penta-acylated species. Among them, three subgroups were detected: the major one compatible with the sugar core bearing two 3-OH 20:0 fatty acids, a 3-OH 13:0 or a 3-OH 12:0 fatty acid, a 27-OH 28:0 fatty acid and one saturated fatty acid. Each signal differs in a C18:0 acyl unit from the corresponding hexa-acylated family. On the other hand, a subgroup bearing one 3-OH 20:0 fatty acid, one 27-OH 28:0 fatty acid and two non-polar fatty acids was shown. A minor subgroup compatible with structures containing two hydroxylated and three non-polar fatty acids was also detected. The results obtained showed that nor-harmane was an excellent matrix for charged lipid A structural studies in both, positive and negative ion modes.

Avaliação físico - química e da textura (instrumental e sensorial) de salsichas elaboradas com diferentes níveis de quitosana - poli - (a 1 4) n acentil d glucosamina / Physico chemical evaluation and instrumental and sensory texture analysis of low fat sausages with different levels of chitosan poly (a 1 4) n acetyl d glucosamine

Fueron evaluados los efectos de la adición de diferentes niveles de quitosana en salchichas tipo frankfurter con bajo contenido de gordura sobre la composición química, rendimiento y perfil de textura instrumental (TPA) y sensorial en relación a la salchicha control (C), sin adición de quitosana y con el mismo porcentaje de gordura. Después del procesamiento las salchichas fueron empacadas al vacío y almacenadas a 4ºC para análisis posterior. Los resultados obtenidos mostraron que la adición de quitosana, con los porcentajes utilizados, no presentaron diferencias significativas (p<0.05) sobre la composición química y rendimiento, cuando comparadas con la salchicha control. El perfil de textura instrumental de la salchicha control con relación a las demás con adición de quitosana, no presentó diferencias significativas (p<0.05) para las características de adesividad, elasticidad y coesividad, mientras que hubo diferencias con relación a la dureza, gomosidad y masticabilidad. El perfil sensorial no mostró diferencias (p<0.05) entre el control y las salchichas con diferentes niveles de quitosana, en todas las propiedades sensoriales evaluadas. La matriz de correlación entre las variables instrumental y sensorial demostró que no hubo correlación entre la dureza sensorial y la dureza instrumental. El coeficiente de correlación sensorial mostró alta correlación entre la elasticidad con la gomosidad, y buena correlación entre la dureza con fracturabilidad, adhesividad y liberación de humedad.

Effects of addition of several concentrations of chitosan to low-fat Frankfurter-type sausage on the chemical composition, yield, Textural (instrumental) and sensory perfil analysis were evaluated. The sausages were processed, vacuum packed and stored at 4ºC. There were no significant differences in chemical composition and yield for the sausages with and without chitosan. According to the textural instrumental analysis there were no significant differences in adhessiveness, elasticity, and cohesiveness. However, there were significant differences for hardness, gumminess and chewiness. The sensory analysis profile was similar for the control and the chitosan containing samples according to all the characteristic evaluated. The correlation matrix between instrumental and sensory analysis showed that there was no correlation between the instrumental and the sensory hardness. The sensory correlation coefficient showed a strong correlation between elasticity with gumminess, and a good correlation between hardness and with fracturability, adhesiveness e moisture loss.

[A comparison of the lipopolysaccharides and O-specific polysaccharides of Azospirillum brasilense Sp245 and its omegon-Km mutants KM018 and Km252]. / Sravnitel'naia kharakteristika lipopolisakharidov i O-spetsificheskikh polisakharidov Azospirillum brasilense Sp245 i ego omegon-Km mutantov KM018 i KM252.

The lipopolysaccharides (LPSs) extracted from the outer membrane of Azospirillum brasilense Sp245 and its Omegon-Km mutants KM018 and KM252 with a hot aqueous solution of phenol were found to differ in the content of carbohydrates, glucosamine, and total phosphorus and in the proportion of octadecenoic and hexadecanoic acids in the lipid moieties of the LPSs. The carbohydrate moieties of the LPSs were heterogeneous in charge. The analysis of the O-specific polysaccharides (O-PSs) of the mutants KM018 and KM252 by gas-liquid chromatography, IR spectroscopy, and NMR spectroscopy showed that they are composed of the same linear pentasugar repeating units-->2)-beta-D-Rhap-(1-->3)-alpha-D-Rhap-(1-->3)-alpha-D-Rhap-(1-->2)- alpha-D-Rhap-(1-->2)-alpha-D-Rhap-(1-->as the O-PSs of the parent strain Sp245. The reported differences in the biological activity of the LPSs of the parent and mutant strains can be due to their different chemical structure.

The envelopes of amphibian oocytes: physiological modifications in Bufo arenarum.

A characterization of the Amphibian Bufo arenarum oocyte envelope is presented. It was made in different functional conditions of the oocyte: 1) when it has been released into the coelomic cavity during ovulation (surrounded by the coelomic envelope, (CE), 2) after it has passed through the oviduct and is deposed (surrounded by the viteline envelope, (VE), and 3) after oocyte activation (surrounded by the fertilization envelope, (FE). The characterization was made by SDS-PAGE followed by staining for protein and glycoproteins. Labeled lectins were used to identify glycosidic residues both in separated components on nitrocellulose membranes or in intact oocytes and embryos. Proteolytic properties of the content of the cortical granules were also analyzed. After SDS-PAGE of CE and VE, a different protein pattern was observed. This is probably due to the activity of a protease present in the pars recta of the oviduct. Comparison of the SDS-PAGE pattern of VE and FE showed a different mobility for one of the glycoproteins, gp75. VE and FE proved to have different sugar residues in their oligosaccharide chains. Mannose residues are only present in gp120 of the three envelopes. N-acetyl-galactosamine residues are present in all of the components, except for gp69 in the FE. Galactose residues are present mainly in gp120 of FE. Lectin-binding assays indicate the presence of glucosamine, galactose and N-acetyl galactosamine residues and the absence (or non-availability) of N-acetyl-glucosamine or fucose residues on the envelopes surface. The cortical granule product (CGP) shows proteolytic activity on gp75 of the VE.

Solid substrate fermentation of Monascus purpureus: growth, carbon balance, and consistency analysis.

Solid substrate fermentation (SSF) of Monascus purpureus on rice is a promising new technology for obtaining natural pigments. However, before attempts can be made at maximizing pigment yield, all significant macroscopic compounds should be assayed. Here, Monascus purpureus has been grown on rice in batch mode, and the evolution of the main components, biomass, residual rice, O(2), CO(2), ethanol, acetic acid, and pigments, have been followed. This set of data, never previously studied for Monascus SSF, allowed both the performance of a macroscopic elemental balance, which accounted for 83-94% of the initial substrate carbon, and a check of data consistency. Standard consistency analysis showed a significant underestimation of the nitrogen fraction of biomass, but it was unable to discriminate the errors in the carbon balance as a result of the simultaneous presence of two gross errors in the system. A simple stoichiometric model in tandem with consistency analysis explained unaccounted carbon as an underestimation of CO(2) and ethanol. Using the simplified method to estimate ethanol, the macroscopic balance accounted for 87-99% of the initial carbon.

Ultra estrutura da matriz extracelular da polpa de dentes humanos hígidos / Ultrastructural study of the extracellular matrix of pulps of healthy human teeth

Considerando o papel fisiológico e biológico da matriz extracelular, propomo-nos a estudar a estrutura da rede de glicosaminoglicanas (GAGs) e proteoglicanas (PGs) no tecido conjuntivo de polpas de dentes molares humanos hígidos de pacientes ao redor de 20 anos. As polpas foram fixadas para microscopia eletrônica de transmissäo em soluçöes de glutaraldeído com vermelho de rutênio e com tetróxido de ósmio reduzido. A rede de GAGs e PGs foi constituída por grânulos de filamentos dispostos numa extensa malha uniforme, contínua e compacta, com nítido inter-relacionamento com a estrutura e posicionamento das fibrilas colágenas e com a superfície de células do tecido conjuntivo

Primary structure of the oligosaccharide chain of lipopeptidophosphoglycan of epimastigote forms of Trypanosoma cruzi.

The lipopeptidophosphoglycan of epimastigote forms of Trypanosoma cruzi is composed of a glycan linked through a non-N-acetylated glucosamine residue to an inositol phosphorylceramide. Using conventional analysis techniques, including 1H, 13C, and 31P NMR spectroscopy and negative ion fast atom bombardment mass spectroscopy, the structure of the carbohydrate-containing part of the molecule is determined as: (Sequence: see text). There is uncertainty as to which 2-O-substituted alpha-D-Manp unit is attached the side chain or whether it is distributed between the two units. Some of the structures lack the Galf side chain. The inositol unit is linked to ceramide via a phosphodiester bridge. The major aliphatic components of the ceramide portion were lignoceric acid and sphinganine.

Host-parasite relationships in the yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9.

The yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9 reduced the amount of alpha-1,3-glucan in its cell wall from 45 to 3% when subcultured in vitro for several years. This strain regained its alpha-1,3-glucan up to 25% of the total cell wall when grown in vivo. A mutant strain of P. brasiliensis Pb9, named IVIC Pb140, reported to have 1,3-mannan instead of alpha-glucan in the cell wall, could not be recovered from experimentally infected animals. The existence of some relationship between the presence of alpha-1,3-glucan in the cell wall of the yeastlike form and the pathogenicity of this fungus is suggested in this report.

Cell wall composition of the yeast and mycelial forms of Paracoccidioides brasiliensis.

Isolation and chemical analyses of the cell walls of the yeast (Y form) and mycelial forms (M form) of Paracoccidioides brasiliensis and Blastomyces dermatitidis revealed that their chemical composition is similar and depends on the form. Lipids, chitin, glucans, and proteins are the main constituents of the cell walls of both forms of these fungi. There is no significant difference in the amount of lipids (5 to 10%) and glucans (36 to 47%) contained by the two forms. In both fungi, the Y form has a larger amount of chitin (37 to 48%) than the M form (7 to 18%), whereas the M form has a larger amount of proteins (24 to 41%) than the Y form (7 to 14%). Several properties of the glucan of P. brasiliensis were studied. Almost all of the glucan in the Y form was soluble in 1 n NaOH, was weakly positive in the periodic acid-Schiff reaction, was not hydrolyzed by snail digestive juice, and had alpha-glycosidic linkage. Glucans of the M form were divided into alkali-soluble (60 to 65%) and alkali-insoluble (35 to 40%) types. The alkali-soluble glucan was similar to that of the Y form; the alkali-insoluble glucan was positive in the periodic acid-Schiff reaction and was hydrolyzed by snail digestive juice.