Fabrication of 1,4-alpha-D-glucan glucanohydrolase holding Gel-Scaffolds using Agar-Agar, a natural polysaccharide and Polyacrylamide, a synthetic organic polymer for continuous liquefaction of starch

1,4-alpha-D-glucan glucanohydrolase is among the most widely used commercial hydrolytic enzymes acting randomly on the glycosidic linkages of starch resulting in its saccharification and liquefaction. Its applicability in different industries can be improved by enhancing its stability and reusability. Therefore, in the present study attempts have been made to enhance the industrial applicability of 1,4-alpha-D-glucan glucanohydrolase from Bacillus subtilis KIBGE-HAR by adapting immobilization technology. The study developed mechanically stable, enzyme containing gel-frameworks using two support matrices including agar-agar, a natural polysaccharide and polyacrylamide gel, a synthetic organic polymer. These catalytic gel-scaffolds were compared with each other in terms of kinetics and stability of entrapped 1,4-α-D-glucan glucanohydrolase. In case of polyacrylamide gel, Km value for immobilized enzyme increased to 7.95 mg/mL, while immobilization in agar-agar resulted in decreased Km value i.e 0.277 mg/mL as compared to free enzyme. It was found that immobilized enzyme showed maximum activity at 70 °C in both the supports as compared to free enzyme having maximum activity at 60 °C. Immobilized 1,4-α-D-glucan glucanohydrolase exhibited no change in optimal pH 7.0 before and after entrapment in polyacrylamide gel and agar-agar. The enzyme containing gel-scaffold was found suitable for repeated batches of starch liquefaction in industrial processes. Agar-agar entrapped 1,4-α-D-glucanglucanohydrolase was capable to degrade starch up to seven repeated operational cycles whereas polyacrylamide entrapped enzyme conserved its activity up to sixth operational cycle.

Eurytrema coelomaticum infection: correlation between parasite burden and impairment of pancreatic exocrine enzyme secretion / Eurytrema coelomaticum: correlação entre carga parasitária e insuficiência pancreática exócrina

Eurytrema coelomaticum is a trematode reported in the pancreatic ducts of ruminants. It is conjectured that may cause disorders in the pancreas, as well as digestive and metabolic processes dependent on them. This study, determined if there is an impairment of exocrine pancreatic function, and correlated it with parasite burden. Pancreas, blood, and fecal samples were collected from 119 bovines at a abattoir. Stool samples were subjected to the gelatin and x-ray film digestion tests (to detect the presence of trypsin in feces). Using blood samples, the following biochemical tests were performed: amylase, lipase, glucose, fructosamine, cholesterol, triglycerides, total protein, albumin, and globulins. Analyses were correlated with pancreatic parasite burden. Cattle with a high parasitic load presented higher incidence of negative tests in both gelatin digestion and x-ray film digestion tests (P < 0.001) when compared to non-parasitized animals and those with a low parasitic load. Changes in those tests only occurred if the parasitemia was moderate or severe. The activity of the amylase and lipase enzymes was significantly higher in animals with low parasitemia (P < 0.05), compared to non-parasitized animals and with a high parasitic burden. In this study, in cases of high parasitemia, negative results were observed in both gelatin and x-ray film in the feces digestion tests. However, the low infection of E. coelomaticum, higher levels of serum amylase and lipase that also indicated loss of pancreatic exocrine functions were reported.

Eurytrema coelomaticum, um trematódeo de ductos pancreáticos de ruminantes. Conjectura-se que possa ocasionar transtornos nas funções pancreáticas, mais especificamente nos processos digestivos e metabólicos dependentes destas. Neste estudo, o objetivo foi determinar se há comprometimento da função pancreática exócrina, correlacionado-a a carga parasitária. Foram utilizados pâncreas e respectivas amostras de sangue e fezes de 119 bovinos. As amostras de fezes foram submetidas aos testes de digestão da gelatina em tubo e digestão de filme radiográfico, ambos para detecção de tripsina nas fezes. Foram realizados os seguintes exames bioquímicos em amostras de sangue: amilase, lipase, glicemia, frutosamina, colesterol, triglicerídeos, proteínas totais, albumina e globulinas. Após isto, as análises bioquímicas foram correlacionadas com a quantidade numérica de parasitas encontrados no pâncreas (post-mortem). Houve maior quantidade de testes negativos (digestão do filme radiográfico e prova de digestão da gelatina) nos animais com alta carga parasitária (P < 0.001), quando comparados aos animais não parasitados e com baixa carga parasitária. Portanto, os exames supracitados se alteram somente se a quantidade de parasitas for moderada ou severa. As atividades das enzimas amilase e lipase foram significativamente maiores nos animais que apresentavam baixa parasitemia (P < 0.05), em comparação com os animais com alta carga parasitária e não parasitados. Conclui-se que em quadros de alta parasitemia há alteração significativa nos testes de digestão nas fezes, e que em quadros de baixa parasitemia há alterações significativas nos valores de amilase e lipase séricas, ambos comprovando alterações pancreáticas importantes, de acordo com o quadro de parasitemia.

Formación de almidón retrogradado relacionado a niveles de amilosa en el almidón de papa (Solanum tuberosum) y chuño / Retrograded starch formation dependent on amylose level of potatoe starch (Solanum tuberosum) and chuño

INTRODUCCIÓN: la papa es el tubérculo más importante producido a nivel mundial como producto alimenticio. Una de las formas ancestrales de conservación de la papa en los países andinos es la obtención de un producto llamado chuño, obtenido a partir de las denominadas papas amargas. Existen pocos datos respecto a la composición química y nutricional del chuño y sobre los cambios producidos en el almidón durante su elaboración en el proceso de congelado-secado en condiciones específicas de temperatura y exposición a rayos ultravioleta. OBJETIVO: el objetivo del trabajo fue realizar una revisión bibliográfica respecto a cambios producidos en la relación de amilosa y amilopectina, la composición de almidón aislado de papa y la modificación de las características de este almidón en el proceso de elaboración de chuño. METODOLOGÍA: La revisión bibliográfica se ha realizado con la recopilación de tres fuentes referenciales de estudios realizados sobre el chuño y su proceso de elaboración, artículos sobre la estructura del almidón de papa y otros tubérculos del mismo género, artículos de otros productos alimenticios del Altiplano boliviano y peruano, y finalmente la influencia de los cambios de la estructura del almidón en el incremento de la formación de almidón retrogradado. RESULTADOS: la revisión bibliográfica realizada, señala que el proceso de elaboración de chuño eleva el porcentaje de amilosa en el contenido total de almidón, lo cual está relacionado a procesos de exposición a radiación UV y a cambios de temperaturas muy drásticos, que van entre -13,5 ºC y 16 ºC. Este proceso llevaría a la activación de enzimas, como amilasas, para la catálisis de reacciones de ruptura de enlaces como principal ruta del proceso; sin embargo, se podrían evaluar otras causas. CONCLUSIONES: el mayor porcentaje de amilosa permitiría la obtención de altos porcentajes de almidón retrogradado.

INTRODUCTION: the potato is the most important tuber produced worldwide as a food product. One of the ancestral ways of preserving pootatoes in the Andean countries is a product called chuño (traditional Andean freeze and sun-dried potato), obtained from the so-called bitter potatoes. There are few data regarding the chemical and nutritional composition of chuño and the changes produced in the starch during its preparation in the freeze-drying process under specific conditions of temperature and exposure to ultraviolet rays. OBJECTIVE: the objective of this article was to do a bibliographic review regarding changes produced in the amylose and amylopectin ratio, the composition of isolated potato starch and the modification of the characteristics of this starch in the process of making chuño. METHODOLOGY: the literature review methodology has the compilation of three reference sources of studies carried out on chuño and its production process, articles about the structure of potato starch and other tubers, research on other food products from the Bolivian and Peruvian of Altiplano and finally the influence of the changes in starch structure in relation to the increasing of retrograde starch formation. RESULTS: the results show that the process of making chuño increases the percentage of amylose in the total starch content, which is related to the processes of exposure to UV radiation and very drastic temperature changes, ranging between -13, 5 ºC and 16 ºC. This process would lead to the activation of enzymes, such as amylases, for the catalysis of bond breaking reactions as the main route of the process; however, other causes could be evaluated. CONCLUSIONS: the higher percentage of amylose would allow the obtaining of high percentages of retrograded starch.

Hallazgos en tomografía computada de las complicaciones de la pancreatitis aguda / Findings of Acute Pancreatitis Complications on CT

Resumen La pancreatitis aguda es una patología inflamatoria del páncreas que se caracteriza por dolor abdominal y elevación de los niveles séricos de amilasa. Desde el punto de vista imagenológico, se la estudia con tomografía computada (TC), siendo esa la modalidad de elección para evaluar la extensión y la localización de las complicaciones. Según la gravedad, el tiempo de evolución y la extensión, hay distintos tipos de complicaciones y una variedad de hallazgos en TC.

Abstract Acute pancreatitis is an inflammatory pathology of the pancreas characterized by abdominal pain and elevated levels of serum amylase. In radiology, it is studied with computed tomography (CT), which is the preferred method to evaluate the extent and location of complications. Depending on the severity, time of evolution, and extent, there are different types of complications and a variety of findings on the CT scan.

Macroamilasemia en el Hospital Víctor Lazarte Echegaray de Trujillo, Perú: reporte de un caso / Macroamylasemia at the Hospital Víctor Lazarte Echegaray in Trujillo (Peru): Case report

Resumen La macroamilasemia debe sospecharse en todo paciente con aumento catalítico de amilasa α plasmática persistente y sin clínica de dolor abdominal, descartando otras etiologías de patología pancreáticas y extrapancreática. La macroamilasemia se caracteriza por la unión de complejos de amilasa α con inmunoglobulina, más frecuente inmunoglobulina A; pueden presentarse 3 tipos de macroamilasemia. Es importante realizar el diagnóstico diferencial de otras patologías que puedan causar el aumento de la amilasa y así evitar los procedimientos innecesarios. Se presenta el caso de un paciente de 53 años, de sexo femenino, que ingresó a emergencia por clínica de dolor abdominal y hiperamilasemia, que fue diagnosticada inicialmente de pancreatitis aguda.

Abstract Macroamylasemia should be suspected in any patient with a persistent catalytic increase of plasma α-amylase but no other clinical signs of abdominal pain after ruling out other causes of pancreatic and extra-pancreatic disease. The binding of α-amylase complexes with immunoglobulin, most commonly immunoglobulin A, characterizes this condition. Macroamylasemia is classified into three kinds. To prevent unnecessary procedures, it is critical to make a differential diagnosis of other conditions that can cause amylase increase. The present article reports the case of a 53-year-old female patient who was admitted to the emergency room with abdominal pain and hyperamylasemia, who was initially diagnosed with acute pancreatitis.

Uso do gérmen de milho desengordurado, com e sem adição de um complexo enzimático, em dietas para cães / [Use of defatted corngerm with and without the addition of an enzyme complex in dog diets]

Objetivou-se avaliar o coeficiente de digestibilidade aparente (CDA) dos nutrientes, a palatabilidade das dietas e as características fecais de cães alimentados com uma dieta controle e uma dieta contendo 20% de gérmen desengordurado (GD), com e sem adição de complexo enzimático (amilase, xilanase, betaglucanase e mananase). Para o experimento de digestibidade e das características fecais, foram utilizados 12 cães adultos, distribuídos em delineamento em blocos ao acaso, em esquema fatorial 2 x 2 (dieta x enzima). O segundo experimento avaliou a palatabilidade, por meio da primeira escolha e da razão de ingestão (RI) da dieta DC vs. 20% de GD, utilizando-se 16 cães. O teste de palatabilidade contou com três dias consecutivos, totalizando 48 repetições. A dieta com inclusão de 20% de GD teve os menores valores de CDA da MS, da EB e da EM (P<0,05). A inclusão do complexo enzimático melhorou o CDA da MS, da EB e da EM (P<0,05). Não foram observadas diferenças nas características fecais (P>0,05). Em relação à palatabilidade, os cães preferiram a dieta 20% de GD, tanto na primeira escolha como na RI (P<0,05). A inclusão de enzimas às dietas melhora a digestibilidade dos nutrientes e da EM, sendo um aditivo com potencial uso na alimentação de cães.(AU)

The objective was to evaluate the apparent digestibility coefficient (ADC) of nutrients, diet palatability and fecal characteristics of dogs fed diets containing degreased germ (DG), and a control diet (DC) - both with and without the addition of enzyme complex (amylase, xylanase, betaglucanase and mananase). For the digestibility and fecal characteristics experiment 12 adult dogs were used, distributed in a randomized block design, in a 2 x 2 factorial scheme (diet x enzyme). The second experiment evaluated palatability using the first choice and ingestion ratio (IR) of DC diet vs. 20%gD, using 16 dogs. The palatability test had three consecutive days, totaling 48 repetitions. The diet with inclusion of 20% DG had the lowest ADC values of DM, GE and ME (P <0.05). Inclusion of the enzyme complex improved ADC of DM, GE and ME (P <0.05). No differences in fecal characteristics were observed (P >0.05). Regarding palatability, dogs preferred the 20% DG diet in both first choice and IR (P <0.05). Inclusion of enzymes in diets improves nutrient digestibility and ME, being an additive with potential use in dog food.(AU)

Abdominal drain amylase on the first day after pancreatectomy: a predictive factor for pancreatic fistula

SUMMARY OBJECTIVE: To analyze abdominal drain on the first postoperative day and evaluate its predictive nature for the diagnosis of Pancreatic Fistula exclusion, seeking to establish a cutoff point from which lower values demonstrate safety in excluding the possibility of this complication. METHODS: From August 2017 to June 2020, data from 48 patients undergoing pancreatic resection were collected and analyzed from a prospective cohort. The patients were divided into two groups, one group consisting of patients who did not develop PF (Group A), and the other composed of patients who developed PF (Group B). The receiver operation characteristic curve was constructed, and cutoff points were evaluated by calculating sensitivity and specificity. RESULTS: Group A brought 30 patients together (62.5%) and Group B brought 18 patients together (37.5%). The 444 U/L value was the most satisfactory cutoff point for the receiver operation characteristic curve (CI 0.690-0.941), with a sensitivity of 94.4% and a specificity of 60%, thus being able to select 18 of 30 patients who did not succumb to PF. CONCLUSIONS: Abdominal drain on the first postoperative day can be used as a predictive factor in the diagnosis of PF exclusion (CI 0.690-0.941), with the value of 444 U/L being the best performance cutoff point.

Screening and Characterization of Bacillus Strains Producing Highly Thermostable Amylase from Various Hot Springs of Algeria

HIGHLIGHTS Screening extremophile Bacillus strains from various Hot Springs Characterization Of Bacillus Strains Producing Highly Thermostable Amylase Genetic identification of the best strains

Abstract Currently thermostable Amylase represents a broad biotechnological interest and desired by a various industries. In this study, forty-six bacterial strains belonging to the genus Bacillus were isolated from various hot springs in the North West of Algeria based on their ability to degrade starch and produce amylase in Starch Agar medium. The majority of isolates showed a positive amylolytic activity. In order to select the most thermostables amylase the effect of temperature on enzymes was estimated, therefore the study of amylase thermostability was culminated by the selection of Four Strains having an interesting optimum of activity and range of stability, reaching 75 °C for the strains HBH1-2, HBH1-3, HBH3-1and 85 °C for the strain HC-2, This indicates that the Enzyme produced by retained strains have optimum activity at high temperature. The identity of the selected strains was established on the basis of the morphological, biochemical characteristics and phylogenetic position as determined by 16S Ribosomal DNA gene sequencing. The whole strains belonged to the genus Bacillus and their phylogeny were also reported in this study.

Ansa pancreática: desafio no diagnóstico etiológico em paciente jovem com pancreatite grave / Ansa pancreatica: etiological diagnosis challenge in a young patient with severe pancreatitis

A ansa pancreática é uma variação anatômica rara dos ductos pancreáticos. Consiste numa comunicação entre o ducto pancreático principal (Wirsung) e o ducto pancreático acessório (Santorini). Recentemente, estudos têm demonstrado estar essa variação anatômica implicada como fator predisponente e significativamente associada a episódios recorrentes de pancreatite aguda. A pancreatite é uma entidade clínica pouco frequente na infância. Diferente dos adultos, as causas mais comuns incluem infecções virais, por ascaris, medicamentosas, traumas e anomalias estruturais. O objetivo deste estudo foi relatar um caso de pancreatite aguda grave não alcoólica e não biliar, em um paciente jovem de 15 anos, em cuja propedêutica imagenológica evidenciou-se alça, comunicando com os ductos pancreáticos ventral e dorsal, compatível com ansa pancreática.

Ansa pancreatica is a rare anatomical variation of the pancreatic ducts. It consists of communication between the main pancreatic duct (Wirsung) and the accessory pancreatic duct (Santorini). Recently, studies have shown that this anatomical variation is implicated as a predisposing factor and significantly associated with recurrent episodes of acute pancreatitis. Pancreatitis is a rare clinical entity in childhood. Different from that in the adults, the most common causes include viral and ascaris infections, drugs, traumas, and structural abnormalities. The objective of this study was to report a case of a severe non-alcoholic and non-biliary acute pancreatitis in a 15-year-old patient, whose propedeutic imaging showed a loop communicating with the ventral and dorsal pancreatic ducts, consistent with ansa pancreatica.

Comparative molecular studies of halophilic bacteria from saline water and soil in the Saudi environment / Estudos moleculares comparativos de bactérias halofílicas de água salina e solo no meio ambiente saudita

Halophilic bacteria are microorganisms that grow optimally in the presence of the very high concentration of sodium chloride. Halophiles are vital sources of various enzymes including hydrolases, which are very stable and catalytically highly efficient at high salt concentration and other extreme conditions such as high temperature, pH and presence of organic solvents. Several hydrolases such as amylases, proteases, and lipases have been obtained from halophilic bacteria and are commonly used for various industrial applications. We initiated a screening to isolate and characterize the halophilic bacteria from the Red Sea, which is one of the saltiest bodies of water in the world. Water and soil samples, collected from the Red Sea coast, Jeddah, Saudi Arabia, were screened for isolation of halophilic bacteria. Ten bacterial isolates were obtained, which were characterized by biochemical tests and 16S rRNA gene sequencing. Hydrolase producing bacteria among the isolates were screened by plate assay on starch and gelatin agar plates for amylase and protease, respectively. Two bacterial isolates i.e. Bacillus glycinifermentans S3 and Enterobacter cloacae W1were found to possess significant amylase and protease activity.

Bactérias halofílicas são microrganismos que crescem de maneira ideal na presença de uma concentração muito alta de cloreto de sódio. Halófilos são fontes vitais de várias enzimas, incluindo hidrolases, que são muito estáveis e cataliticamente altamente eficientes em alta concentração de sal e outras condições extremas, como alta temperatura, pH e presença de solventes orgânicos. Várias hidrolases como amilases, proteases e lipases foram obtidas a partir de bactérias halofílicas e são comumente usadas para várias aplicações industriais. Iniciamos uma triagem para isolar e caracterizar as bactérias halofílicas do Mar Vermelho, que é um dos corpos de água mais salgados do mundo. Amostras de água e solo, coletadas na costa do Mar Vermelho, Jeddah, na Arábia Saudita, foram examinadas quanto ao isolamento de bactérias halofílicas. Foram obtidos dez isolados bacterianos, caracterizados por testes bioquímicos e seqüenciamento do gene 16S rRNA. As bactérias produtoras de hidrolase entre os isolados foram triadas por ensaio em placa em placas de amido e ágar de gelatina para amilase e protease, respectivamente. Verificou-se que dois isolados bacterianos, isto é, Bacillus glycinifermentans S3 e Enterobacter cloacae W1, possuíam significativa atividade de amilase e protease.

Chemical characterization of the antioxidant, antihyperglycemic and antihypertensive capacities of red rice (Oryza sativa L. ) whole flour / Caracteristicas físico-químicas, capacidad antioxidante, anti-hiperlucémica y antihipertensiva de la harina de arroz rojo (Oryza sativa L. )

Oryza sativa L. rice has large amounts of proteins and minerals, besides presenting several pigmented varieties. Red rice is distinguishable due to its great nutritional value compared to the regular white variety. Its red pericarp pigmentation is due to the bioactive compounds that are responsible for its health benefits. The objective of this study was to evaluate the physical-chemical characterization, antioxidant, antihyperglycemic and antihypertensive capacity of flours of three different red rice cultures (Rubi, Virgínia and Pequeno). All samples presented specific levels of carbohydrates for cereals with low fat content and excellent levels of protein and resistant starch. In addition, the samples had a high antioxidant, antihyperglycemic and antihypertensive capacity. Antihyperglycemic capacities were measured as percent inhibition for amylase (56.7-76.5%) and glycosidase (81.0-76.6%), respectively, and antihypertensive capacity as the percentage inhibition of the angiotensin converting enzyme (38.4-34.7%). In addition, Pequeno flour presented the best results for antioxidant and antihyperglycemic capacity in comparison to the two flours tested. Thus, all red rice flours can be a source of functional compounds when added to food.

El arroz integral (Oryza sativa L.) posee importantes cantidades de proteínas, vitaminas, minerales y fitoquímicos. El arroz rojo se destaca por su gran valor nutricional. La pigmentación roja del pericarpio está asociado al contenido de compuestos bioactivos, que están directamente relacionados a los beneficios de salud humana. Teniendo en cuenta lo antes expuesto se propuso evaluar las caracteristicas físico-químicas, capacidad antioxidante, anti-hiperglucémica y antihipertensiva de las harinas de tres diferentes cultivos de arroz rojo (Rubí, Virginia y Pequeño). Todas las muestras presentaron niveles específicos de carbohidratos para cereales con bajo contenido de grasa y altos contenidos de proteína y almidón resistente. Además, las muestras presentaron una alta capacidad antioxidante, anti-hiperglucémica y antihipertensiva. La capacidad anti-hiperglicémica se midió en porcentaje de inhibidores de α-amilasa (56.7-76.5%) y α-glucosidasa (81.0-76.6%), respectivamente; y capacidad antihipertensiva como el porcentaje de inhibición de la enzima convertidora de la angiotensina (38.4-34.7%). El cultivar Pequeño presentó mayor capacidad antioxidante y anti-hiperglucémica en comparación a los demás cultivares. Así, todas las harinas de arroz rojo pueden ser vehículos de compuestos funcionales en los alimentos.

Diversity and antimicrobial activity of culturable fungi associated with sea anemone Anthopleura xanthogrammica

Background: The main objective of this study was to isolate fungi associated with Anthopleura xanthogrammica and measure their antimicrobial and enzymatic activities. A total of 93 fungal strains associated with A. xanthogrammica were isolated in this study, of which 32 isolates were identified using both morphological characteristics and internal transcribed spacer (ITS) sequence analysis. The antibacterial activities of 32 fungal isolates were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Edwardsiella tarda, Vibrio harveyi, Fusarium oxysporum, and Pyricularia oryzae by agar diffusion assay. Extracellular hydrolytic enzyme activities of the fungal isolates were determined by agar diffusion assays. Enzyme activities were detected from clear halo size. Results: The isolated fungi belonged to 18 genera within 7 taxonomic orders of 1 phylum. The genera Aspergillaceae were the most diverse and common. The antimicrobial activities of 32 isolates were evaluated, and 19 (59.4%) of fungi isolate displayed unique antimicrobial activities. All fungal strains displayed at least one enzyme activity. The most common enzyme activities in the fungi isolates were amylase and protease, while the least common were pectinase and xylanase. Conclusions: This is first report on the sea anemone-derived fungi with antimicrobial and enzyme activities. Results indicated that sea anemone is a hot spot of fungal diversity and a rich resource of bioactive natural products.

Independent and synergistic activity of the flavonoids of Gracilaria corticata as promising antidiabetic agents

The therapeutic approaches for Type 2 Diabetes Mellitus rely most on the usage of oral hypoglycaemic drugs. These drugs have adverse side effects and hence alternative medicines are continuously explored. The present study intends to investigate the antidiabetic potential of the flavonoids present in Gracilaria corticata. The flavonoids were isolated (FEGC) and their inhibitory activity on the carbohydrate hydrolysing enzymes such as α-amylase and α-glucosidase was analysed. The flavonoids were found to inhibit α-amylase and α-glucosidase with an IC50 value of 302 µg and 75 µg respectively. The synergistic effect of FEGC and luteolin was also investigated and the results show that both FEGC and luteolin inhibited synergistically at half their IC50 values. The observations of this study reveal that the flavonoids of G. corticata have potential antidiabetic activity and can act independently or synergistically in the management of Type 2 Diabetes Mellitus

Amylase from Bacillus sp. Produced by Solid State Fermentation Using Cassava Bagasse as Starch Source

Abstract Amylases are enzymes involved in starch hydrolysis, generating the most diverse products, such as maltose, glucose and dextrins. This work aimed the study of the production of amylolytic enzymes via solid-state fermentation (SSF) using "crueira", an essentially starchy cassava residue, as substrate-support and Bacillus sp. as microorganism. For the implementation of the experimental part, a Central Composite Design (CCD) with three variables (initial moisture, pH and temperature) was made. Each test was examined at 24, 48 and 72 hours by the method of starch dextrinizing activity. The optimum production conditions were 60% initial moisture, pH 6 and 37 °C. The maximum yield was 437.76 U/g in 72 hours of fermentation. The optimum temperature of enzyme performance was 65 °C. The pH optimum range was 4 to 6. The Co2 +, Ca2 + and K+ ions positively influenced the activity of enzymes and the Fe2+ ion had no effect on enzymatic activity. On the other hand, the ions Hg2+, Zn2+, Cu2+, Mn2+ and Mg2+ adversely influenced enzymatic activity. Therefore, producing amylases from Bacillus sp. and using crueira as a substrate is possible.

The Effect of Thymosin Alpha 1 on Severe Acute Pancreatitis in Rats / Estudios sobre el efecto de la timosina alfa 1 sobre la pancreatitis aguda grave en ratas

ABSTRACT Objective: To observe the effect of thymosin alpha l (Tα1) on severe acute pancreatitis (SAP) in rats. Methods: Twenty-four adult male Sprague-Dawley rats were randomly divided into three groups (eight in each group): control group (Group A), SAP group (Group B) and Tα1 treatment group (Group C). Animal models of SAP were made by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. Rats in Group C were treated with Tα1 (6 mg/kg) via intraperitoneal administration prior to SAP modelling. Eight rats in each group were sacrificed at 12 hours, respectively, after modelling. The serum levels of amylase, tumour necrosis factor-α (TNF-α), interleukin-lβ (IL-lβ and interleukin-6 (IL-6) were detected in each group. The pathological scores of the tissue in the pancreas head were observed by light microscopy. Results: The levels of serum amylase of Group B were 6378 ± 538 U/L, which were significantly higher than those (4587 ± 478 U/L) of Group C (p < 0.05). The levels of serum TNF-α of Group B were 360.32 ± 28.67 pg/mL, which were higher than those (269.99 ± 26.11 pg/mL) of Group C (p < 0.05). The levels of serum IL-lβ of Group B were 435.93 ± 36.00 pg/mL, which were higher than those (312.42 ± 17.89 pg/mL) of Group C (p < 0.05). The levels of serum IL-6 of Group B were 433.90 ± 28.36 pg/mL, which were higher than those (289.98 ± 23.00 pg/mL) of Group C (p < 0.05). The pancreatic pathological scores of Group B were 13.34 ± 2.19, which were higher than those (6.39 ± 1.86) of Group C (p < 0.05). Conclusion: Thymosin alpha 1 could decrease proinflammatory cytokines and reduce pancreas injury and had a protective effect in rats with SAP. This provides a new strategy for the clinical treatment of SAP.

RESUMEN Objetivo: Observar el efecto de la timosina alfa l (Tα1) sobre la pancreatitis aguda grave (PAG) en ratas. Métodos: Veinticuatro ratas Sprague-Dawley adultas machos fueron divididas aleatoriamente en tres grupos (ocho en cada grupo): grupo de control (grupo A), grupo de PAG (grupo B) y grupo de tratamiento con Tα1 (grupo C). Los modelos animales de PAG fueron creados mediante inyección retrógrada de taurocolato de sodio al 5% en el conducto biliopancreático. Las ratas del grupo C se trataron con Tα1 (6 mg/kg) via administración intraperitoneal antes del modelado de PAG. Las ocho ratas en cada grupo fueron sacrificadas a las 12 horas, respectivamente, después del modelado. Los niveles séricos de amilasa, factor-α de necrosis tumoral (TNF-α), interleucina-β (Il-β) e interleucina-6 (IL-6) fueron detectados en cada grupo. Las puntuaciones patológicas del tejido en la cabeza del páncreas fueron observadas mediante microscopía de luz. Resultados: Los niveles de amilasa sérica del grupo B fueron 6378 ± 538 U/L, y resultaron significativamente más altos (p < 0.05) que los niveles 4587 ± 478 U/L del grupo C. Los niveles séricos de TNF-α del grupo B fueron 360.32 ± 28.67 pg/mL, y resultaron ser más altos (p < 0.05) que los 269.99 ± 26.11 pg/mL del grupo C. Los niveles séricos de Il-β del grupo B fueron 435.93 ± 36.00 pg/mL, y fueron más altos (p < 0.05) que los 312.42 ± 17.89 pg/mL) del grupo C. Los niveles de suero IL-6 del grupo B fueron 433.90 ± 28.36 pg/mL, y resultaron ser más altos (p < 0.05) que los 289.98 ± 23.00 pg/mL del grupo C. Las puntuaciones patológicas pancreáticas del grupo B fueron 13.34 ± 2.19, y resultaron ser más altas (p < 0.05) que las puntuaciones 6.39 ± 1.86 del grupo C. Conclusión: La timosina alfa pudo disminuir las citoquinas proinflamatorias y reducir la lesión del páncreas, y tuvo un efecto protector en las ratas con PAG. Esto ofrece una nueva estrategia para el tratamiento clínico de PAG.

Effects of baicalin on inflammatory reaction, oxidative stress and PKDl and NF-kB protein expressions in rats with severe acute pancreatitis

Abstract Purpose: To investigate the effects of baicalin on inflammatory reaction, oxidative stress and protein kinase D1 (PKD1) and nuclear factor-kappa B (NF-κB) protein expressions in severe acute pancreatitis (SAP) rats. Methods: Sixty rats were divided into sham operation, model, and low-, medium- and high-dose baicalin group. SAP model was established in later 4 groups. The later 3 groups were injected with 0.1, 0.2 and 0.4 ml/100 g 5% baicalin injection, respectively. At 12 h, the serum SAP related indexes and inflammatory factors, peripheral blood CD3 and γδT cell percentages, wet/dry ratio and pancreas ascites volume, oxidative stress indexes and PKD1 and NF-κB protein expressions in pancreatic tissue were determined. Results: Compared with model group, in high-dose baicalin group the wet/dry ratio and ascites volume, serum amylase level, phospholipase A2 activity, TNF-α, IL-1 and IL-6 levels, and pancreatic malondialdehyde level and PKD1 and NF-κB protein expression were significantly decreased (P < 0.05), and peripheral blood CD3 and γδT cell percentages and pancreatic superoxide dismutase and glutathione peroxidase levels were significantly increased (P < 0.05). Conclusion: Baicalin can resist the inflammatory reaction and oxidative stress, and down-regulate protein kinase D1 and nuclear factor-kappa B protein expressions, thus exerting the protective effects on severe acute pancreatitis in rats.

O valor da amilase, obtido precocemente nos drenos abdominais, é teste útil em prever a ocorrência de fístula pancreática após duodenopancreatectomia: lições aprendidas de um centro do Sul do Brasil / Early drain fluid amylase is useful to predict pancreatic fistula after pancreatoduodenectomy: lessons learned from a southern brazilian center

ABSTRACT BACKGROUND: Pancreatic fistula represents the most feared complication after pancreatoduodenectomies, being the major responsible for the high morbidity and mortality after this operation. Its incidence remains around 10% to 30%. In recent years, several authors have studied the value of amylase in abdominal drains fluid, measured at an early stage after the surgical procedure, as a useful tool to identify patients at risk of developing pancreatic fistula. OBJECTIVE: To analyze the value of early drain fluid amylase as a method to predict the occurrence and severity of postoperative pancreatic fistula in patients undergoing pancreatoduodenectomies. METHODS: We evaluated 102 prospective patients submitted to pancreatoduodenectomies from January 2013 to June 2017. The mensuration of amylase in abdominal drains was performed on days 1, 3, 5 and 7 in all patients. Patients were divided into three groups according to postoperative day 1 (POD1) results: values <270 U/L (group 1); between 271 and 5.000 U/L (group 2); and values >5.000 U/L (group 3). RESULTS: The incidence of pancreatic fistula was 25.5%, being 3.33%, 27.3% and 41.02% in the three groups, respectively. Compared with group 1, the risk of developing pancreatic fistula increased with increasing amylase values on POD1. Amylase values on POD1 and POD3 of patients with pancreatic fistula were higher than in the other ones without this complication (P<0.001). In addition, in group 3, 37.5% of patients with pancreatic fistula evolved to death (P<0.001). Finally, in this group, patients who died had drain fluid amylase values on POD1 significantly higher than the others in the same group (P<0.001). CONCLUSION: Early drain fluid amylase value is a useful test to stratify patients in relation to the risk of developing pancreatic fistula after pancreatoduodenectomies, in addition to correlate with the severity of this complication.

RESUMO CONTEXTO: A fístula pancreática representa a complicação mais temida após as duodenopancreatectomias, sendo a grande responsável pela elevada morbi-mortalidade após esta operação. Sua incidência permanece em torno de 10% a 30%. Nos últimos anos, diversos trabalhos têm estudado o valor da amilase nos drenos abdominais, medido de forma precoce após o procedimento cirúrgico, como ferramenta útil para a identificação dos pacientes sob risco de desenvolver fístula pancreática. OBJETIVO: Avaliar o valor da amilase no fluido dos drenos abdominais, obtido precocemente no pós-operatório, como método para prever a ocorrência e severidade da fístula pancreática nos pacientes submetidos a duodenopancreatectomias. MÉTODOS: Foram avaliados 102 pacientes prospectivos submetidos a duodenopancreatectomias no período de janeiro de 2013 a junho de 2017. A dosagem da amilase nos drenos abdominais foi realizada nos dias 1, 3, 5 e 7 em todos os pacientes. Os pacientes foram divididos em três grupos conforme os resultados do 1o PO: valores <270 U/L (grupo 1); entre 271 e 5.000 U/L (grupo 2); e valores >5.000 U/L (grupo 3). RESULTADOS: A incidência de fístula pancreática foi de 25,5%, sendo 3,33%, 27,3% e 41,02% nos três grupos, respectivamente. Comparados ao grupo 1, o risco de desenvolver fístula pancreática foi crescente com o aumento da amilase no 1o PO. Os valores das amilases no 1o PO e 3o PO dos pacientes com fístula pancreática foram maiores do que nos pacientes sem essa complicação (P<0,001). Além disso, no grupo 3, 37,5% dos pacientes com fístula pancreática evoluíram para óbito (P<0,001). Por fim, neste grupo, os pacientes que evoluíram para óbito tiveram valores de amilase no 1o PO significativamente maiores do que os demais pacientes (P<0,001). CONCLUSÃO: O valor da amilase, medido de forma precoce nos drenos abdominais no pós-operatório de duodenopancreatectomias, é teste útil para estratificar pacientes em relação ao risco de apresentar fístula pancreática, além de se correlacionar com a severidade dessa complicação.

Utilização dos níveis de amilase no dreno abdominal para diagnóstico de fístulas anastomóticas após o bypass gástrico em Y de Roux / Drain amylase levels may indicate gastrojejunostomy leaks after roux-en-y gastric bypass

ABSTRACT BACKGROUND: Although the incidence of leaks after Roux-en-Y gastric bypass (RYGB) significantly decreased over time, their detection still remains challenging. OBJECTIVE: This study aimed to determine the usefulness of drain amylase levels to detect leaks after RYGB. METHODS: This is a population-based study which enrolled 170 individuals who underwent RYGB. Drain amylase levels were determined on the first and fourth postoperative days. Two thresholds were evaluated: three times higher than the serum levels (parameter I) and higher than 250 IU/L (parameter II). The main outcomes evaluated were perioperative morbidity, the occurrence of leaks, 30-day readmissions and reoperations, hospital stay, and mortality. RESULTS: Considering the parameter I, high drain amylase levels were significantly associated with leaks (12.5% vs 0; P<0.00001). Considering the parameter II, high drain amylase levels were significantly associated with longer hospital stay (8±5.7 vs 4.5±1.3 days; P=0.00032), 30-day reoperations (50% vs 3%; P=0.000285), and leaks (50% vs 0; P<0.00001). The parameter I presented a sensitivity of 100% and specificity of 95.9%, whereas the parameter II presented a sensitivity of 100% and a specificity of 99.4%. CONCLUSION: The determination of drain amylase levels after RYGB was a significant indicator of leaks, hospital stay, and 30-day reoperations. This finding reinforces the importance of abdominal drainage in the RYGB within this context.

RESUMO CONTEÚDO: Embora a incidência de fistulas após o bypass gástrico em Y de Roux (BGRY) tenha diminuído significativamente com a evolução da técnica, sua detecção continua desafiadora. OBJETIVO: Determinar a acurácia dos níveis de amilase no dreno abdominal para detector fístulas após o BGYR. MÉTODOS: Este é um estudo populacional que avaliou 170 indivíduos submetidos ao BGYR. Os níveis de amilase no dreno foram determinados no primeiro e quarto dias de pós-operatório. Dois pontos de corte foram avaliados: três vezes maior que os níveis séricos (parâmetro I) e acima de 250 UI/L (parâmetro II). Os principais desfechos estudados foram: morbidade perioperatória, ocorrência de fístulas, reinternações e reoperações nos primeiros 30 dias, permanência hospitalar e mortalidade. RESULTADOS: Considerando o parâmetro I, altos níveis de amilase do dreno foram indicadores significativos de fístulas (12,5% vs 0; P<0,00001). Considerando o parâmetro II, altos níveis de amilse no dreno estiveram significativamente associados a maior permanência hospitalar (8±5,7 vs 4,5±1,3 dias; P=0,00032), frequência de reoperações (50% vs 3%; P=0,000285) e ocorrência de fístulas (50% vs 0; P<0,00001). O parâmetro I apresentou sensibilidade de 100% e especificidade de 95,9%, enquanto o parâmetro II apresentou sensibilidade de 100% e especificidade de 99,4%. CONCLUSÃO: A determinação dos níveis de amilase no dreno após o BGYR foi um indicador significativo de fístulas, permanência hospitalar e reoperações. Este achado reforça a importância da drenagem abdominal no BGYR dentro deste contexto.

Filamentous fungi producing enzymes under fermentation in cassava liquid waste / Fungos filamentosos produtores de enzimas sob fermentação em manipueira

The conversion of agroindustrial residues by microorganisms has been explored from fermentative processes to obtain several bioactive molecules. The objective of this work was to isolate and select filamentous fungi present in cassava liquid waste for the production of amylase, carboxymethylcellulose (CMCase), pectinase and xylanase using the same residue as induction substrate in fermentative processes. A total of 65 filamentous fungi were isolated and qualitative tests indicated that approximately 86% of these strains were able to produce at least one of the enzymes and 32% capable of producing the four enzymes. Fermentation assays in cassava liquid residue-containing medium showed 6 fungal lines as potential enzyme producers. The maximum activities of pectinase, xylanase, amylase and CMCase were respectively observed at 96 hours of fermentation by the strain by the strain Aspergillus sp. B5C; at 120 hours (163.6 ± 0.13 nKat mL-1), by Aspergillus sp. B4I; at 144 hours (99.8 ± 0.24 nKat mL-1), by Penicillium sp. B3A; and at 48 hours (55.5 ± 0.21 nKat mL-1), by Aspergillus sp. B4O. These results suggest that cassava liquid waste was source of filamentous fungi producing amylase, CMCase, pectinase and xylanase, as well as a promising alternative substrate for bioprocesses aiming the production of enzymes.

A conversão de resíduos agroindustriais por micro-organismos tem sido explorada a partir de processos fermentativos para obtenção de diversas moléculas bioativas. O objetivo deste trabalho foi isolar e selecionar fungos filamentosos presentes em manipueira para produção de amilase, carboximetilcelulase (CMCase), pectinase e xilanase utilizando o próprio resíduo como substrato indutor. Um total de 65 fungos filamentosos foi isolado e testes qualitativos indicaram que, aproximadamente, 86% dessas linhagens foram hábeis em produzir pelo menos uma das enzimas e 32% capazes de produzir as quatro enzimas. Ensaios fermentativos em meio contendo manipueira apontaram 6 linhagens fúngicas como potenciais produtoras de enzimas. As atividades máximas de pectinase, xilanase, amilase e CMCase foram observadas, respectivamente, às 96 horas de fermentação (67.4 ± 0,6 nKat mL-1), pela linhagem Aspergillus sp. B5C; às 120 horas (163.6 ± 0,13 nKat mL-1), por Aspergillus sp. B4I; às 144 horas (99.8 ± 0,24 nKat mL-1), por Penicillium sp. B3A; e às 48 horas (55.5 ± 0,21 nKat mL-1), por Aspergillus sp. B4O. Estes resultados sugerem a manipueira como fonte de fungos filamentosos produtores de amilase, CMCase, pectinase e xilanase, além de um promissor substrato alternativo para bioprocessos visando a produção dessas enzimas.

Effect of ultraviolet radiation on physiological and biochemical properties of yeast Saccharomyces cerevisiae during fermentation of ultradispersed starch raw material

Background: Study of correlation between pretreatment of yeast with ultraviolet radiation and efficiency of further fermentation of wort made of ultrafine grain particles to ethanol. Results: We investigated three races of industrial yeast Saccharomyces cerevisiae (native and irradiated by ultraviolet). Physiological properties during fermentation of starchy wort were tested in all variants. It was shown that activation of the yeast by ultraviolet radiation allows to further increase the ethanol yield by 25% on average compared with the native yeast races when using thin (up to micro- and nano-sized particles) or standard grain grinding. Conclusions: Using mechanical two-stage grinding of starchy raw materials and ultraviolet pretreatment of yeast, the efficiency of saccharification of starch and fermentation of wort to ethanol was increased.