Sucrose, Lactose, and Xylitol Exposures Affect Biofilm Formation of Streptococcus mutans

Abstract Objective: To determine the level of biofilm formation of S. mutans after being exposed to 5% sucrose, 8% lactose, or 1% xylitol. Material and Methods: This research was a laboratory-based experimental study with post-test only control group design. S. mutans was grown in test tubes containing tryptose soy broth (TSB) medium supplemented with 1% glucose. They were incubated at 37° C for 24 hours to grow the biofilms. The culture was then exposed to 5% sucrose, 8% lactose or 1% xylitol, incubated for 24 hours at 37° C, and examined using ELISA at a wavelength of 625 nm. The statistical analysis was performed using a one-way analysis of variance followed by the least significant difference test (a=0.05). Results: There were some differences in the biofilm formation of S. mutans after exposure to 5% sucrose, 8% lactose, or 1% xylitol (p<0.05). An LSD test indicated significant differences among the biofilm formations after exposure to 5% sucrose and 8% lactose and between 5% sucrose and 1% xylitol. In comparison, there were no significant differences (p>0.05) between 8% lactose and 1% xylitol. Conclusion: Sucrose, lactose and xylitol can form biofilms and the formation of lactose biofilms is the same as xylitol.

Development of a new protocol for freeze-drying preservation of Pseudoalteromonas nigrifaciens and its protective effect on other marine bacteria

Background: Freeze-drying is known as one of the best methods to preserve bacterial strains. Protectant is the key factor affecting the survival rate of freeze-dried strains. In addition, salinity, bacterial suspension concentration, drying time, and other factors can also affect the survival rate of strains to varying degrees. At present, there are relatively few studies on freeze-drying preservation of marine bacteria. In the present study, we performed the freeze-drying protectant screening and optimized the preservation conditions for Pseudoalteromonas nigrifaciens, which is widely distributed in marine environment. The protective effects of the screened protectants were verified by 18 other marine bacterial strains. Results: The results indicated that the combination of 5.0% (w/v) lactose, 5.0% (w/v) mannitol, 5.0% (w/v) trehalose, 10.0% (w/v) skim milk powder, 0.5% (w/v) ascorbic acid and 0.5% (w/v) gelatin was the best choice for the preservation of P. nigrifaciens. The suggested salinity and concentration of initial cell suspension were 10 g/L NaCl and 1.0 × 109 CFU/mL, respectively. Furthermore, stationary-phase cells were the best choice for the freeze-drying process. The highest survival rate of P. nigrifaciens reached 52.8% when using 5­10% (w/v) skim milk as rehydration medium. Moreover, the other 18 marine strains belonging to Pseudoalteromonas, Vibrio, Photobacterium, Planomicrobium, Edwardsiella, Enterococcus, Bacillus, and Saccharomyces were freezedried under the abovementioned conditions. Their survival rates were 2.3­95.1%. Conclusion: Collectively, our results supported that the protectant mixture and parameters were beneficial for lyophilization of marine bacteria

Low FODMAPs diet for functional abdominal pain disorders in children: critical review of current knowledge / Dieta com baixo teor de FODMAPs para distúrbios de dor abdominal funcional em crianças: revisão crítica do conhecimento atual

ABSTRACT Objective: This narrative review aimed to provide practitioners a synthesis of the current knowledge on the role of a low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet in reducing symptoms associated with functional abdominal pain disorders in children. This review is focused on the pathophysiology, efficacy and criticism of low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet in children. Sources: Cochrane Database, Pubmed and Embase were searched using specific terms for Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet interventions and functional abdominal pain disorders. Summary of the findings: In children, only one Randomized Control Trial and one open-label study reported positive results of low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols diet; one Randomized Control Trial showed exacerbation of symptoms with fructans in children with Irritable Bowel Syndrome; no effect was found for the lactose-free diet whilst fructose-restricted diets were effective in 5/6 studies. Conclusions: In children there are few trials evaluating low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols in functional abdominal pain disorders, with encouraging data on the therapeutic efficacy particularly of fructose-restricted diet. Additional efforts are still needed to fill this research gap and clarify the most efficient way for tailoring dietary restrictions based on the patient's tolerance and/or identification of potential biomarkers of low Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols efficacy, to maintain nutritional adequacy and to simplify the adherence to diet by labeling Fermentable Oligosaccharides Disaccharides Monosaccharides and Polyols content in commercial products.

RESUMO Objetivo: Nos últimos anos, foram feitos esforços consideráveis para esclarecer o papel da dieta com baixo teor de oligossacarídeos fermentáveis, dissacarídeos, monossacarídeos e polióis (FODMAPs) para o tratamento de distúrbios gastrintestinais funcionais (DGIFs). Esta revisão narrativa teve como objetivo fornecer aos profissionais uma síntese do conhecimento atual sobre o papel de uma dieta com baixo teor de FODMAPs (BFM) na redução dos sintomas associados a distúrbios funcionais de dor abdominal (DFDA) em crianças. Esta revisão está focada na fisiopatologia, eficácia e crítica da dieta BFM em crianças. Fontes: O banco de dados Cochrane, Pubmed e Embase foram pesquisados com o uso dos termos específicos para intervenções na dieta FODMAP e DFDA. Resumo dos achados: Em crianças, apenas um estudo controlado randomizado e um estudo aberto relataram resultados positivos da dieta BFM; um estudo controlado randomizado mostrou exacerbação dos sintomas com frutanos em crianças com síndrome do intestino irritável; nenhum efeito foi encontrado para a dieta livre de lactose, enquanto dietas com restrição de frutose foram eficazes em 5/6 estudos. Conclusões: Existem poucos estudos que avaliam BFM em DFDA em crianças, com dados encorajadores sobre a eficácia terapêutica, particularmente de dietas com restrição de frutose. Esforços adicionais ainda são necessários para preencher essa lacuna de pesquisa e esclarecer a maneira mais eficiente de adaptar as restrições dietéticas com base na tolerância do paciente e/ou identificação de biomarcadores potenciais de eficácia da BFM, para manter a adequação nutricional e simplificar a adesão à dieta, ao incluir informações sobre conteúdo de FODMAPs em rótulos de produtos comerciais.

Dieta baja en oligosacáridos, disacáridos, monosacáridos y polioles fermentables y calidad de vida en pacientes con síndrome de intestino irritable en Colombia / Diet low in fermentable oligosaccharides, disaccharides, monosaccharides and polyols, and quality of life in patients with irritable bowel syndrome in Colombia

Resumen Introducción. Cada vez hay más información sobre la efectividad de una dieta baja en oligosacáridos, disacáridos, monosacáridos y polioles fermentables (Fermentable Oligosaccharides, Disaccharides, Monosaccharides and Polyols, FODMAP) en pacientes con síndrome de intestino irritable. Sin embargo, no hay estudios que estimen la eficacia de esta estrategia en nuestro medio. Objetivo. Determinar el efecto de una dieta baja en FODMAP en la calidad de vida y en la intensidad de los síntomas en pacientes con síndrome de intestino irritable en Colombia. Materiales y métodos. Se recopiló información clínica y demográfica de todos los pacientes en el momento de su inclusión; después, un encuestador entrenado utilizó la encuesta Irritable Bowel Syndrome Quality of Life (IBS-QoL) para estimar la calidad de vida de los pacientes. La intensidad de los síntomas se evaluó mediante una escala visual, antes y después del inicio de la dieta baja en FODMAP. Resultados. La muestra final incluyó a 50 personas. Se observó una reducción significativa de todos los síntomas (reducción promedio de todas las escalas: 19,8 mm; IC95%: 16,2-23,4 mm; p<0,001), y un incremento en todas las escalas de la IBS-QoL, incluida la de resumen global de la encuesta (14,7 puntos; IC95%: 9,4-20,1; p<0,001). El sexo, la edad, el índice de masa corporal, el estado socioeconómico y el régimen de salud, no se asociaron con la mejoría de la calidad de vida relacionada con la salud. Conclusión. La dieta supervisada y baja en FODMAP disminuyó los síntomas y mejoró la calidad de vida en pacientes con síndrome de intestino irritable. Es necesario hacer estudios controlados sobre otros factores ligados a la evolución del síndrome para confirmar estos resultados.

Abstract Introduction: A growing body of evidence has pointed out the effectiveness of a diet low in fermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAP) in patients with irritable bowel syndrome. However, there are no local studies to estimate the effects of this strategy on the symptoms and the health-related quality of life in these patients in Colombia or Latin America. Objective: To determine the effect of a diet low in FODMAP on the quality of life and the severity of symptoms in patients with irritable bowel syndrome in Colombia. Materials and methods: We collected clinical and demographic information of all patients at the time of inclusion. Immediately afterwards, a trained interviewer applied the IBS-QoL survey to estimate the quality of life of patients. Then, we evaluated the intensity of the symptoms using an analogue visual scale, before and after the diet low in FODMAP. Results: We included 50 subjects in the final analysis. We observed an increase in all the IBS-QoL scales (average increase in overall summary: 14.7 points, 95% CI: 9.4 to 20.1; p<0.001) and a significant reduction in all symptoms (-19.8 mm; 95% CI: 23.4 mm 16.2 mm; p<0.001). Sex, age, body mass index, socioeconomic status and the health care provider were not associated with the improvement in the health-related quality of life. Conclusion: A low diet in FODMAP reduced symptoms and improved quality of life in Colombian patients with irritable bowel syndrome. Controlled studies taking into account other factors linked to the severity of irritable bowel syndrome are required.

Fermentation optimization and enzyme characterization of a new ι-Carrageenase from Pseudoalteromonas carrageenovora ASY5

Background: A new ι-carrageenase-producing strain was screened from mangroves and authenticated as Pseudoalteromonas carrageenovora ASY5 in our laboratory. The potential application of this new strain was evaluated. Results: Medium compositions and culturing conditions in shaking flask fermentation were firstly optimized by single-factor experiment. ι-Carrageenase activity increased from 0.34 U/mL to 1.08 U/mL after test optimization. Optimal fermentation conditions were 20°C, pH 7.0, incubation time of 40 h, 15 g/L NaCl, 1.5% (w/v) yeast extract as nitrogen source, and 0.9% (w/v) ι-carrageenan as carbon source. Then, the crude ι-carrageenase was characterized. The optimum temperature and pH of the ι-carrageenase were 40°C and 8.0, respectively. The enzymatic activity at 35­40°C for 45 min retained more than 40% of the maximum activity. Meanwhile, The ι-carrageenase was inhibited by the addition of 1 mmol/L Cd2+ and Fe3+ but increased by the addition of 1 mmol/L Ag+, Ba2+, Ca2+, Co2+, Mn2+, Zn2+, Fe2+, and Al3+. The structure of oligosaccharides derived from ι-carrageenan was detected using electrospray ionization mass spectrometry (ESI-MS). The ι-carrageenase degraded ι-carrageenan, yielding disaccharides and tetrasaccharides as main products. Conclusions: The discovery and study of new ι-carrageenases are beneficial not only for the production of ι-carrageenan oligosaccharides but also for the further utilization in industrial production.

Identificación diferenciación de monosacárido y disacáridos diasteroméricos no derivatizados por esi-it-ms/ms / Identification nnd differentiation of underivatzed diasteromeric mono-and disaaccharides by esi-it-ms/ms

Los azúcares son importantes moléculas que desempeñan funciones trascendentales de señalización celular en los organismos superiores. Su complejidad estructural, representada por sus isómeros, anómeros y diasterómeros, amerita la implementación de metodologías modernas, rápidas y sensibles para su identificación y diferenciación. La espectrometría de masas y su analizador de trampa de iones brinda nuevas alternativas de análisis que favorecen el control de las energías de fragmentación de los analitos. A pesar de que la diferenciación de estereoisómeros no ha sido el campo de aplicación principal de la espectrometría de masas, se ha implementado una metodología para diferenciar los monosácaridos β-D-galactosa y β-D-glucosa y los disacαridos β-D-galactopiranosil-(1→4)-β-D-glucopiranσsido (lactosa), α-D-glucopiranosil-(1→4)-β-D-glucopiranσsido (maltosa) y β-D-fructofuranosil-(2↔1)-α-D-glucopiranσsido (sacarosa) a travιs de sus aductos con amonio y litio por ESI-IT-MS/MS en infusión directa. Se emplean diferentes energías de fragmentación para asegurar la existencia de iones marcadores de la estereoquímica de los analitos. Se evidencia que controlar las energías de colisiones en el análisis estructural de moléculas provee una poderosa y moderna herramienta analítica para los laboratorios de análisis.

Sugars are important molecules with remarkable cell signals pathway functions in higher organisms. The structural complexity of sugar represented by its isomeric, anomeric and diasteromeric configurations deserve the implementation of modern, rapid and sensitive methodologies for its identification and differentiation. Mass spectrometry and its analyzer of ion trap provide new alternative techniques that encourage the control of the fragmentation energies supplied to molecules. Since stereoisomer differentiation is consider outside the mass spectrometry domain, a methodology has been applied in order to differentiate β-D-galactose, β-D-glucose and the disaccharides β-D-galactopyranosyl-(1→4)-β-D-glucopyranoside (lactose), α-D-glucopyranosyl-(1→4)-β-D-glucopyranoside (maltose) y β-D-fructofuranosyl-(2↔1)-α-D-glucopyranoside (sacarose) trough its ammonium and lithium adducts by infusion on ESI-IT-MS/MS mass spectrometer. Different fragmentation energies have been used to ensure the ion marker occurrence in the analyte stereochemistry. It is evident that the collision energies control in structural analysis of molecules provides a powerful and modern analytical tool to be applied in control laboratories.

Determination of unsaturated disaccharides produced from chondroitin sulfate in urine using high performance liquid chromatography with fluorometric post column derivatization / 中国地方病学杂志

Objective To explore the effectiveness of the unsaturated disaccharides derived from chondroitin sulfate(CS)in urine using post column derivation fluorometric method of high performance liquid chromatography (HPLC).Methods Urine samples were prepared with the centrifugal flirtation and the routine precipitation methods separately,then digest the sample with CS lyses,use the post column derivation method of HPLC to fluorometric determine the unsaturated disaecharides(△Di-0S,△Di-4S and △Di-6S),compare the recovery differences of the CS standard in the urine pretreated by the two methods.Results The HPLC system ran well,the lowest detection limit of the method was 10 ng.When the urine was pretreated by the centrifugal flirtation method,the recovery of △Di-0S,△Di-4S and △Di-6S produced from the CS standard were 86.93%,87.28%and 85.03%,respectively,while the recovery of the three disaccharides in routine precipitation method were 75.09%,72.96%and 77.70%merely.Conclusions The HPLC post column derivatization fluorometric method is comparatively sensitive,it can be used to determine unsaturated disaccharides decomposed by CS.In the pretreatment procedure of the urine,the centrifugal flirtation method was superior to the routine precipitation method,it can be used as microdetermination for the unsaturated disaccharides.

Computer modelling studies on the modes of binding of some of the α- glycosidically linked disaccharides to concanavalin A.

The possible modes of binding of kojibiose, nigerose, maltose and ManPα(1 →2)Man to concanavalin A have been investigated using computer modelling studies. While α12 linked disaccharides bind to concanavalin A in two modes, i.e. by placing the reducing as well as non-reducing sugar units in the sugar binding site, nigerose or maltose can bind only in one mode, i.e. by placing the non-reducing sugar unit in the binding site. Though, both the sugar residues in α 12 linked disaccharides can reach the binding site, the preference is high for the non-reducing unit. When the non-reducing residue, in any of these disaccharides, enters the binding site, the allowed orientations and the possible hydrogen bonds with the protein seem to be independent of the glycosidic linkage. However, the number of hydrogen bonds the outward sugar residue forms with the protein are dependent on the type of linkage. Atleast one of the hydroxyl groups adjacent to the glycosidic linkage on the outward sugar residue is involved in the formation of a hydrogen bond with the protein suggesting the presence of an extended binding site. The orientation of the reducing sugar residue in the extended binding site is dependent on the linkage. Its orientation in nigerose is flipped when compared to that found in kojibiose or maltose leading to different non-covalent interactions with the protein which affect their binding affinities.