ABSTRACT
OBJECTIVE@#To characterize the bacteriocin-like inhibitory-substances (BLIS) activity of Streptococcus macedonicus MBF10-2 [named BLIS(MBF10-2)], a bacteriocinogenic strain isolated from an Indonesian tofu byproduct.@*METHODS@#BLIS(MBF10-2) was obtained by culturing the bacterium, and standard deferred antagonism assays were used to demonstrate its activity. The antibacterial testing of fractions collected by filtration using 3-30 kDa cut-off membrane sizes were carried out by performing well diffusion method.@*RESULTS@#The growth of Micrococcus luteus, Streptococcus pyogenes, Lactococcus lactis, Leuconostoc mesenteroides and Weissella confusa were inhibited by BLIS(MBF10-2). Interestingly, BLIS-containing fractions obtained from sequential application on ultrafiltration membranes indicated that this bacterium Streptococcus macedonicus MBF10-2 could produces at least two antimicrobial peptides activities, one of which is likely to be a lantibiotic peptide. Potential synergistic activity against certain Gram-positive (but not Gram-negative) species when partnered with antibiotics (ampicillin, tetracycline or kanamycin) were observed.@*CONCLUSION@#Combination of some BLIS(MBF10-2) active fractions with antibiotics (ampicillin, tetracycline or kanamycin) could demonstrate synergistic activities against certain Gram-positive species.
ABSTRACT
Objective To characterize the bacteriocin-like inhibitory-substances (BLIS) activity of Streptococcus macedonicus MBF10-2 [named BLIS(MBF10-2)], a bacteriocinogenic strain isolated from an Indonesian tofu byproduct. Methods BLIS(MBF10-2) was obtained by culturing the bacterium, and standard deferred antagonism assays were used to demonstrate its activity. The antibacterial testing of fractions collected by filtration using 3–30 kDa cut-off membrane sizes were carried out by performing well diffusion method. Results The growth of Micrococcus luteus, Streptococcus pyogenes, Lactococcus lactis, Leuconostoc mesenteroides and Weissella confusa were inhibited by BLIS(MBF10-2). Interestingly, BLIS-containing fractions obtained from sequential application on ultrafiltration membranes indicated that this bacterium Streptococcus macedonicus MBF10-2 could produces at least two antimicrobial peptides activities, one of which is likely to be a lantibiotic peptide. Potential synergistic activity against certain Gram-positive (but not Gram-negative) species when partnered with antibiotics (ampicillin, tetracycline or kanamycin) were observed. Conclusion Combination of some BLIS(MBF10-2) active fractions with antibiotics (ampicillin, tetracycline or kanamycin) could demonstrate synergistic activities against certain Gram-positive species.
ABSTRACT
La harina de nuez de Barinas (Caryodendron orinocense K.) de acuerdo a estudios realizados presenta un contenido de pro teínas en el rango de 15-18%, que permite se pueda considerar como fuente de proteína. Sin embargo no se conoce la composición de estas proteínas. En este estudio se realizó un proceso de fraccionamiento de acuerdo a la solubilidad en diferentes solventes (agua, cloruro de sodio al 5%, hidróxido de sodio 0,02N, etanol al 70%), y el establecimiento de la composición proteica según el peso molecular de la harina de nuez de Barinas obtenida a partir de las nueces secas, molidas y des grasadas, determinando el rango de pesos moleculares por SDS-PAGE y su comparación con el rango de pesos moleculares de las proteínas de la harina de soya (Glycine max). Asi mismo, se determinó la digestibilidad in vitro por hidrólisis enzimática. Los resultados indican un rango de PM de 20.000- 97.000 daltons para la soya y de 6.500-45.000 daltons para la nuez de Barinas. El peso molecular más bajo sugiere una más fácil digestión de las proteínas, que corrobora el valor de la di - ges tibilidad obtenido de 75%. Las proteínas presentes están cons tituidas por albúminas 50,72%, globulinas 15,56%, prolaminas 23,10% y glutelinas 2,52% que representan el 92,15% del total de proteínas presentes. Estas fracciones presentaron una pureza de 81,57%, 94,01%, 70,86% y 92,53%, respectivamente. Estos resultados sugieren la posibilidad de uso de la harina de Caryodendron orinocense K en el desarrollo de productos alimenticios para niños y ancianos.
Nuez de Barinas (Caryodendron orinocense K.) flour has been reported to have a content of proteins in the range of 15-18% that allows considering it as a source of proteins. However, there is no evidence on the composition of these proteins. The objective of this study was the extraction and separation of the different fractions of proteins based on their solubility in different solvents (water, 5% sodium chloride, sodium hydroxide 0.02N, and 70% ethanol), and the protein composition assessment of the nuez de Barinas flour from dried, milled, and defatted nuts; by analysis of the range of mo lecular weight by SDS-PAGE. Molecular weights of proteins from soybean (Glycine max) flour, were used for comparison. In vitro protein digestibility was determined by enzymatic hydrolysis. Results presented soy proteins in the range of 20.000-97.000 daltons while the nuez de Barinas flour proteins were in the range of 6.500-45.000 daltons. Low molecular weight proteins should suggest a much easier digestibility of these proteins, which is related to the digestibility index of 75% found. Protein composition was found to be 50,72% albumins, 15,56% globulins, 23,10% prolamins, and 2,52% glutelins which represent 92,15% of total proteins present. These fractions showed a percent purity of 81.57, 94.01, 70.86 and 92.53, respectively. These results suggest the possibility to use this flour Caryodendron orinocense K. in developing food products for children and elderly persons.
Subject(s)
Humans , Male , Female , Child, Preschool , Child , Aged , Aged, 80 and over , Plants/classification , Proteins/therapeutic use , Albumins , Prolamins , Public Health , Globulins , Nuts/classificationABSTRACT
This report describes a preliminary characterization of proteolytic activity of proteins isolated from lysate of Giardia trophozoites of an axenic Brazilian strain. Fractions obtained by high-performance liquid chromatography (FPLC) were tested in SDS-polyacrylamide gel for the protein profiles, and the proteases activity was analyzed using gelatin impregnated SDS-PAGE. The proteases characterization was based on inhibition assays employing synthetic inhibitors for cysteine (E-64, IAA), serine (PMSF, TPCK, TLCK, and elastatinal), metalo (EDTA) and aspartic (pepstatin) proteases. Among thirty eluted fractions, polypeptide bands were observed in eight of them, however, proteolytic activity was detected in four ones (F23, F24, F25 and F26). Protein profiles of these fractions showed a banding pattern composed by few bands distributed in the migration region of 45 to < 18 kDa. The zymograms revealed proteolytic activity in all the four fractions assayed, mainly distributed in the migration region of 62 to 35 kDa. Among the profiles, the main pronounced zones of proteolysis were distinguished at 62, 55, 53, 50, 46 and 40 kDa. In inhibition assays, the protease activities were significantly inhibited by cysteine (E-64) and serine proteases (TPCK, TLCK and elastatinal) inhibitors. Gels incubated with other cysteine and serine protease inhibitors, IAA and PMSF, respectively, showed a decrease in the intensity of hydrolysis zones. Indeed, in the assays with the inhibitors EDTA for metalloproteases and pepstatin for aspartic proteases, none inhibition was detected against the substrate. These observations are relevants, especially if we consider that to define the real role of the proteases in host-parasite interaction, the purification of these enzymes for detailed studies may be warranted.
O presente estudo consiste em uma caracterização preliminar da atividade proteolítica de frações de proteínas purificadas a partir de lisados de trofozoítos de cepa isolada e axenizada no Brasil. Frações obtidas por cromatografia líquida (FPLC) foram analisadas quanto ao perfil eletroforético em géis de poliacrilamida (SDS-PAGE) e a atividade proteolítica foi avaliada em géis contendo gelatina como substrato. A caracterização das enzimas foi realizada a partir da análise do efeito de inibidores sintéticos de cisteína-proteases (E-64, IAA), serina-proteases (PMSF), serina e cisteína-proteases (TPCK, TLCK, elastatinal), metalo-proteases (EDTA) e aspartil proteases (pepstatina) sobre a degradação do substrato. Entre 30 frações eluídas, bandas de proteínas foram observadas em oito delas, entretanto, atividade proteolítica foi detectada apenas nas frações 23, 24, 25 e 26. O perfil eletroforético das proteínas revelou poucas bandas distribuídas na faixa de 45 a 18 kDa. Os zimogramas revelaram zonas de proteólise na faixa de aproximadamente 62 a 35 kDa, entretanto destacaram-se as bandas de hidrólise de 62, 55, 53, 50, 46 e 40 kDa. Nos ensaios de inibição, a proteólise foi marcantemente inibida por E-64, TPCK, TLCK e elastatinal. Redução discreta da proteólise foi observada com IAA e PMSF, enquanto que EDTA e pepstatina não promoveram alteração dos perfis de hidrólise. Estas observações são relevantes, especialmente se considerarmos que para elucidar o envolvimento das proteases na relação parasita-hospedeiro, a purificação dessas moléculas é um requisito importante.
Subject(s)
Animals , Humans , Giardia/enzymology , Peptide Hydrolases/metabolism , Protozoan Proteins/metabolism , Chromatography, Liquid , Electrophoresis, Polyacrylamide Gel , Peptide Hydrolases/isolation & purification , Protozoan Proteins/isolation & purificationABSTRACT
<p><b>OBJECTIVE</b>This study examined the levels of serum protein fractionation, immunoglobulin (Ig) and antistreptolysin O (ASO) of people (91 males and 84 females aged 10-68 years) living in the Terai region of southern Nepal, as there has previously been no information available about them.</p><p><b>METHODS</b>Blood samples were collected early in the morning after overnight fasting. Serum protein fractionation was carried out by cellulose-acetate electrophoresis. IgG, IgA, IgM and ASO were measured by immuno-turbidimetry with clinical kits.</p><p><b>RESULTS</b>The mean proportion of albumin (Alb) was rather low due to increased globulin (Glb). The Alb level of males was significantly higher than that of females, while the male γ-Glb level was significantly lower than that of females. The mean values of IgG, IgM and ASO for males were lower than those values for females, although the difference was significant only for the IgM value. The mean values of IgM for female age groups of 30-39 years or less were significantly higher than those for the corresponding male age groups. Age correlated positively with IgA, and negatively with ASO in both sexes. γ-Glb and IgG correlated significantly with TP, AlB, α(1)-Glb and IgA in both sexes. ASO correlated with β-Glb and IgA in males, and with γ-Glb and IgG in females.</p><p><b>CONCLUSIONS</b>The fact that the level of γ-Glb, a major component of serum globulin, was high suggested exposure to a highly bacterial and viral environment. These results point to the need to prevent infectious diseases as well as improve their nutritional status, especially for children and young adults.</p>
ABSTRACT
Objective: This study examined the levels of serum protein fractionation, immunoglobulin (Ig) and antistreptolysin O (ASO) of people (91 males and 84 females aged 10−68 years) living in the Terai region of southern Nepal, as there had previously been no information available about them. Methods: Blood samples were collected early in the morning after overnight fasting. Serum protein fractionation was carried out by cellulose-acetate electrophoresis. IgG, IgA, IgM and ASO were measured by immuno-turbidimetry with clinical kits. Results: The mean proportion of albumin (Alb) was rather low due to increased globulin (Glb). The Alb level of males was significantly higher than that of females, while the male γ-Glb level was significantly lower than that of females. The mean values of IgG, IgM and ASO for males were lower than those values for females, although the difference was significant only for the IgM value. The mean values of IgM for female age groups of 30-39 years or less were significantly higher than those for the corresponding male age groups. Age correlated positively with IgA, and negatively with ASO in both sexes. γ-Glb and IgG correlated significantly with TP, Alb, α1-Glb and IgA in both sexes. ASO correlated with β-Glb and IgA in males, and with γ-Glb and IgG in females. Conclusions: The fact that the level of γ-Glb, a major component of serum globulin, was high suggested exposure to a highly bacterial and viral environment. These results point to the need to prevent infectious diseases as well as improve their nutritional status, especially for children and young adults.
Subject(s)
NepalABSTRACT
A hydrocele is a common condition which affects all age-group males including babies. It may be defined as a collection of serous fluid within the tunica vaginalis either surrounding the testis or along the spermatic cord or both. Congenital hydrocele is due to complete failure of the processus funicularis closure and most of the fluid collections subside spontaneously during the first year of life. The great majority of hydrocele are primary variety and its etiology is unknown. Secondary hydrocele may be developed to local injury, epididymitis and testicular neoplasms. Chronic hydrocele is common in tropical and subtropical countries where there is a high .incidence of filariasis. Some investigators have reported that hydrocele fluid collections are due to lymphatic defect or peritoneal fluid, It is generally known that hydrocele fluid resembles blood plasma. We performed electrophoresis and electrolytic analysis of hydrocele fluid and serum in 20 cases of hydrocele patients from January, 1981 to September, 1982. And then we obtained the statistical difference between hydrocele fluid and serum. The results are summarized as below: 1. Total protein of hydrocele fluid (4.125 gm/dl) is lower than that of serum (7,535 gm/dl). Statistical difference is greatly significant (p=below 0.005). 2. The proportion of albumin (71.780%) is higher than that of serum (65.685%). The proportion of alpha 2 and gamma globulin (6.880%, 5.535%) are lower than that of serum (10.125%, 8.730%). These statistical difference are greatly significant (p=below 0.005). 3. The proportion of alpha-1 and beta globulin (3.495%, 12.215%) are higher than that of serum (3.105%, 11.935%). There is no statistical significance (p=above 0.1). 4. Sugar of hydrocele fluid (87.65 mg/dl) is lower than that of serum (95.40 mg/dl), but there is no statistical significance (p=above 0.1). 5. Electrolytes of hydrocele fluid are lower than those of serum. Sodium (130 mEq/1); great statistical significance (p=below 0.005). Potassium (3.93 mEq/l); no statistical significance (p=above 0.1). Chloride (97.95 mEq/l); no statistical significance (p=above 0.1).