Phytochemical analysis and biological activities of ethanolic extract of Curcuma longa rhizome / Análise fitoquímica e atividades biológicas do extrato etanólico do rizoma de Curcuma longa

Abstract Curcuma longa is an important dietary plant which possess several pharmacological activities, including antioxidant, antimicrobial, anti-inflamatory, anticancer and anti clotting etc. The aim of the present study was to determine the phenolic profile of Curcuma longa and in vitro antioxidant and antidiabetic activities. In HPLC chromatogram of Curcuma longa rhizome extract 15 phenolic compounds were identified namely Digalloyl-hexoside, Caffeic acid hexoside, Curdione, Coumaric, Caffeic acid, Sinapic acid, Qurecetin-3-D-galactoside, Casuarinin, Bisdemethoxycurcumin, Curcuminol, Demethoxycurcumin, and Isorhamnetin, Valoneic acid bilactone, Curcumin, Curcumin-O-glucuronide respectively. The ethanolic extract displayed an IC50 value of 37.1±0.3 µg/ml against alpha glucosidase. The IC50 value of DPPH radical scavenging activity was 27.2 ± 1.1 μg/mL. It is concluded that ethanolic extract of Curcuma long is rich source of curcumin and contain several important phenolics. The in vitro antioxidant and alpha glucosidase inhibitory effect of the plant justifies its popular use in traditional medicine.

Resumo A Curcuma longa é uma importante planta presente na dieta da população, pois possui diversas atividades farmacológicas, incluindo antioxidante, antimicrobiana, anti-inflamatória, anticancerígena, anticoagulante etc. O objetivo do presente estudo foi elucidar o perfil fenólico da Curcuma longa e determinar as atividades antioxidante e antidiabética in vitro do extrato. No cromatograma por HPLC do extrato de rizoma de Curcuma longa, foram identificados 15 compostos fenólicos: digaloil-hexosídeo, hexosídeo de ácido cafeico, curdiona, cumárico, ácido cafeico, ácido sinápico, quercetina-3-D-galactosídeo, casuarinina, bisdemetoxicurcumina, curcuminol, demetoxicurcumina, isoramnetina, bilactona de ácido valônico, curcumina e curcumina-O-glicuronídeo. O extrato etanólico apresentou um valor de IC50 de 37,1 ± 0,3 µg / mL em relação à alfa-glucosidase. O valor de IC50 da atividade de eliminação de radicais DPPH foi de 27,2 ± 1,1 μg / mL. Conclui-se que o extrato etanólico de Curcuma longa é uma rica fonte de curcumina e contém várias substâncias fenólicas importantes. O efeito antioxidante in vitro e inibidor da alfa-glucosidase da planta justifica seu uso popular na medicina tradicional.

Effect observation and literature review of enzyme replacement therapy in late-onset Pompe disease / 中华神经科杂志

Objective:To analyze the efficacy and safety of enzyme therapy in late-onset Pompe disease (LOPD) patients, so as to provide reference for the treatment and prognosis of LOPD.Methods:The effect of α-glucosidase (GAA) on a patient diagnosed with LOPD in the Affiliated Hospital of Jining Medical University was observed and analyzed. Besides, literature related to enzyme therapy in LOPD patients were searched in PubMed, Web of Science, Medline databases. Twenty-one studies containing clinical data from 910 LOPD patients related to enzyme therapy were finally included for analysis.Results:The patient developed muscle weakness since he was 16 years old. The GAA activity in peripheral blood was 0. Electromyography suggested myogenic lesions in both lower extremities. Compound heterozygous mutations of GAA gene were found by next- generation sequencing. Muscle biopsy revealed characteristic vacuolar changes. After eight years of diagnosis, the patient was given enzyme therapy for 18.5 months, 20 times in total. The symptoms of muscle weakness were slightly improved in the early stages of treatment without obvious adverse reactions. Most of the 910 LOPD patients were stabilized or had improved muscular and (or) respiratory function following treatment with GAA.Conclusion:GAA treatment is effective and well tolerated. In patients with advanced severe LOPD, enzyme replacement therapy remains effective even years after onset.

Enfermedad de Pompe: detección de portadores en un estado del sureste de México: reporte de un caso / Pompe disease: carrier detection in a southeastern state of Mexico: a case report

Resumen ANTECEDENTES: La enfermedad de Pompe es un padecimiento autosómico recesivo del metabolismo del glucógeno causado por la deficiencia parcial o total de la enzima lisosomal alfa-glucosidasa ácida (GAA). Su incidencia es variable y depende de la región geográfica y el origen étnico. OBJETIVO: Reportar un caso de mutación previamente descrita en población mexicana relacionada con enfermedad de Pompe. CASO CLÍNICO: Paciente de 21 años, llegó al servicio de Genética enviada de un centro de salud por el antecedente de dos muertes perinatales. En la evaluación de la pareja se encontraron antecedentes de importancia de consanguinidad (primos hermanos) y ginecoobstétricos: dos embarazos, dos partos y dos muertes perinatales por miocardiopatía de causa desconocida. Las ecografías de los embarazos previos no reportaron alteraciones, tampoco en la exploración física. Debido a los antecedentes se decidió la búsqueda de portadores de la enfermedad de Pompe en la rama materna. CONCLUSIÓN: Se tomaron en consideración la detección y diagnóstico de portadores de errores innatos del metabolismo debido a los antecedentes recopilados en la historia clínica (muertes perinatales, consanguinidad). Se reporta una mutación previamente descrita en población mexicana relacionada con la enfermedad de Pompe.

Abstract BACKGROUND: Pompe's disease is an autosomal recessive disease of glycogen metabolism; caused by partial or total deficiency of the lysosomal enzyme alpha-glucosidase acid (GAA). Its incidence is variable and depends on the geographical region and ethnicity. OBJECTIVE: Report a case of previously described mutation in Mexican population related to Pompe disease. CLINICAL CASE: A 21-year-old female who attends first-time genetics services, with reference to a health center due to a history of two perinatal deaths. An evaluation of the couple is performed, with a history of the importance of consanguinity (first cousins), a gynecoobstetric history: pregnancy 2, births 2. Two perinatal deaths due to cardiomyopathy of unknown cause. Ultrasound findings of previous pregnancies, without report of alterations. A physical examination of both patients, without phenotypic alteration. Due to the aforementioned background, it is decided to search for carriers of Pompe disease in the maternal branch. CONCLUSION: In this case the detection and diagnosis of carriers of inborn errors of metabolism, the antecedents collected in the clinical history (perinatal deaths, consanguinity) were taken into consideration. Publicizing a mutation previously described in the Mexican population, which is related to Pompe disease.

Isolation and activity evaluation of endophytic fungi from Glycyrrhiza uralensis / 中草药

Objective: To isolate and purify endophytic fungus from Glycyrrhiza uralensis, and screen the specific strain with better antibacterial and antioxidant activity (DPPH• radical scavenging, total reducing power, determination of hydroxyl radical scavenging) and anti-alpha-glucosidase activity. Methods: The endophytic fungi were isolated and purified from G. uralensis by tissue cutting. N-butanol, ethyl acetate and ethanol were used to extract the fermentation liquid and mycelium. Antibacterial activity was detected by filter paper. Three methods were used to characterize antioxidant activity. A total of 36 endophytic fungi were isolated from G. uralensis by PNPG method. Results: A total of seven genera were isolated from 108 samples by concentrated fermentation liquid extraction. Trichoderma. sp was the dominant species. The experimental results showed that 43.52% of the samples had different degrees of antibacterial effect, of which 8.33% performed well; The extracts showed different levels of antioxidant activity, of which 4.63% to 10.12% showed better performance in the three methods. 99.07% of the samples had different levels of anti-α-glucosidase activity, of which 5.56% of the samples performed well. Conclusion: The strains isolated from G. uralensis have good antibacterial activity, antioxidant activity, and anti-alpha-glucosidase activity, which further study is needed.

Inhibitory Effect of the Stem Bark Extracts of Albizia chevalieri Harms on the Activity of Alpha-glucosidase Obtained from Aspergillus niger

The aim of the current study is to evaluate the inhibition of α-glucosidase activity by stem bark extract of Albizia chevalieri. The activity of alpha glucosidase was assayed in vitro using 50 mM acetate buffer pH 6.0 (prepared from acetic acid and sodium acetate) and various concentration of maltose (0.5 mM to 10 mM). Five test tubes, labeled TA – TE, each containing 1.5 ml of acetate buffer, 0.5 ml of alpha glucosidase and 0.5 ml of a known concentration of plant extract and control tubes (CA – CE) were assessed for Alpha glucosidase activity. The results showed that hexane, ethyl acetate and methanol extracts inhibited α-glucosidase activity. The results further indicated that the extracts act by competitive inhibition with inhibition constant of 232 mg/ml, 157 mg/ml and 67 mg/ml for hexane, ethyl acetate and methanol extracts, respectively. The value for the inhibition constants shows that there is a strong binding of the enzyme to the inhibitor as the polarity of solvent increases. The inhibitory activity of Albizia chevalieri may be due one or more of the phytochemicals present in the extracts.

Pseudodeficiency alleles affect the newborn screening of glycogen storage disease typeⅡ / 中华检验医学杂志

Objective To investigate the effect of pseudodeficiency alleles on the newborn screening of glycogen storage disease typeⅡ(GSDⅡ) by using afluorometric enzymatic assay to determine acidα-glucosidase (GAA) activity in dried blood spot (DBS). Methods A total of 30507 newborns' DBSs, obtained from Newborn Screening Center of Xinhua Hospital Shanghai Jiao Tong University School of Medicine from May to December 2017, were screened for GSDⅡby fluorometric enzymatic assay of GAA activity. The suspected positive DBSs after the first and second screening were directly analyzed by Sanger sequencing of GAA to confirm the diagnosis. Retrospective analysis of 3172 controls without GSDⅡand 36 GSDⅡpatients were conducted to investigate the carrier status of pseudodeficiency alleles. Statistical analysis of frequency of pseudodeficiency alleles were carried out by Chi-square test or Fisher exact probability test. Results GAA activity of 30507 newborns showed a positively skewed distribution. Twenty-nine cases of newborns, suspected to be GSDⅡwere confirmed to be normal with genetic analysis of the original DBSs. Among the 29 suspected positive cases, 24 cases were homozygous for pseudodeficiency alleles c. [1726A/A; 2065A/A], and the other 5 cases were c. [1726G/A; 2065G/A] heterozygote. The frequency of c. 1726G>Ahomozygote in 3172 non-GSDⅡcontrols was 2.08％(66/3172), and c. 1726G>A homozygote occurred in allelic conjunction with c. 2065G>Ahomozygote. Frequency of c. [1726A; 2065A] haplotype in 3172 controls was 3.2％(206/6344). Frequency of c. [1726A/A;2065A/A] homozygote in 36 GSDⅡpatients (16.67％, 6/36) was significantly higher than that in non-GSDⅡcontrols(2.08％, 66/3172) (χ2=34.517, P<0.001). Conclusions Pseudodeficiency alleles show a high frequency in Chinese, which leads to a high false positive rate in the newborns screening of GSDⅡ.The afterword genetic analysis of the original DBS after the GAA activity screening could reduce the effect of pseudodeficiency alleles on the newborns screening of GSDⅡ.

Pseudodeficiency alleles affect the newborn screening of glycogen storage disease typeⅡ / 中华检验医学杂志

Objective@#To investigate the effect of pseudodeficiency alleles on the newborn screening of glycogen storage disease type Ⅱ(GSDⅡ) by using afluorometric enzymatic assay to determine acid α-glucosidase (GAA) activity in dried blood spot (DBS).@*Methods@#A total of 30 507 newborns′ DBSs, obtained from Newborn Screening Center of Xinhua Hospital Shanghai Jiao Tong University School of Medicine from May to December 2017, were screened for GSD Ⅱ by fluorometric enzymatic assay of GAA activity. The suspected positive DBSs after the first and second screening were directly analyzed by Sanger sequencing of GAA to confirm the diagnosis. Retrospective analysis of 3 172 controls without GSDⅡand 36 GSD Ⅱ patients were conducted to investigate the carrier status of pseudodeficiency alleles. Statistical analysis of frequency of pseudodeficiency alleles were carried out by Chi-square test or Fisher exact probability test.@*Results@#GAA activity of 30 507 newborns showed a positively skewed distribution.Twenty-nine cases of newborns, suspected to be GSDⅡwere confirmed to be normal with genetic analysis of the original DBSs. Among the 29 suspected positive cases, 24 cases were homozygous for pseudodeficiency alleles c.[1726A/A; 2065A/A], and the other 5 cases were c.[1726G/A; 2065G/A] heterozygote. The frequency of c.1726G>Ahomozygote in 3 172 non-GSD Ⅱcontrols was 2.08% (66/3 172), and c.1726G>A homozygote occurred in allelic conjunction with c.2065G>Ahomozygote. Frequency of c.[1726A; 2065A] haplotype in 3 172 controls was 3.2%(206/6 344). Frequency of c.[1726A/A; 2065A/A] homozygote in 36 GSDⅡpatients (16.67%, 6/36) was significantly higher than that in non-GSD Ⅱcontrols(2.08%, 66/3 172) (χ2=34.517, P<0.001).@*Conclusions@#Pseudodeficiency alleles show a high frequency in Chinese, which leads to a high false positive rate in the newborns screening of GSDⅡ.The afterword genetic analysis of the original DBS after the GAA activity screening could reduce the effect of pseudodeficiency alleles on the newborns screening of GSDⅡ.

Identification of Seven Novel Mutations in the Acid Alpha-glucosidase Gene in Five Chinese Patients with Late-onset Pompe Disease / 中华医学杂志(英文版)

<p><b>Background</b>Pompe disease is a rare lysosomal glycogen storage disorder linked to the acid alpha-glucosidase gene (GAA). A wide clinical and genetic variability exists between patients from different ethnic populations, and the genotype-phenotype correlations are still not well understood. The aim of this study was to report the clinicopathological and genetic characteristics of five Chinese patients with late-onset Pompe disease (LOPD) who carried novel GAA gene mutations.</p><p><b>Methods</b>Clinical and pathological data of patients diagnosed with glycogen storage disease at our institution from April 1986 to August 2017 were collected, and next-generation sequencing of frozen muscle specimens was conducted.</p><p><b>Results</b>Of the five patients included in the study, the median disease onset age was 13 years, with a median 5 years delay in diagnosis. The patients mainly manifested as progressive weakness in the proximal and axial muscles, while one patient developed respiratory insufficiency that required artificial ventilation. In muscle biopsies, vacuoles with variable sizes and shapes appeared inside muscle fibers, and they stained positive for both periodic acid-Schiff and acid phosphatase staining. Ten GAA gene mutations, including seven novel ones (c.796C>A, c.1057C>T, c.1201C>A, c.1780C>T, c.1799G>C, c.2051C>A, c.2235dupG), were identified by genetic tests.</p><p><b>Conclusions</b>The seven novel GAA gene mutations revealed in this study broaden the genetic spectrum of LOPD and highlight the genetic heterogeneity in Chinese LOPD patients.</p>

Evaluation of antiparasitic, anticancer, antimicrobial and hypoglycemic properties of organic extracts from Panamanian mangrove plants

Objective:To investigate 33 organic extracts of mangrove plants for: antiparasitic, anticancer, and antibacterial activities, as well as their ability to inhibit the activity of the α-glucosidase enzyme.Methods:Leaves from all different plant mangrove species located in five mangrove zones of the Pacific coast of Panama were collected according to standard procedures. Qualitative phytochemical analysis of the organic extracts was performed by thin layer chromatography. The antiparasitic activity against Plasmodium falciparum, Trypanosoma cruzi and Leishmania donovani, toxicity against Artemia salina, anticancer activity in MCF-7 cell line, and antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa of all organic extract were investigated according protocols stablished in our institution. Finally, the ability to inhibit the enzymatic activity of α-glucosidase was evaluated by monitoring the hydrolysis of p-nitrophenyl α-D- glucopyranoside.Results:Thirty-three different samples belonging to nine different species of vascular plants with seeds of true mangroves were collected. Triterpenoids, phenolics, and tannins were the main groups of compounds found in the sampled mangroves. Saponins, quinones, and coumarins were found in less than 50% of the samples. Laguncularia racemosa showed moderate activity against Plasmodium falciparum. None of the extracts presented anticancer activity. Rhizophora mangle exhibited potent activity against Staphylococcus aureus and Bacillus subtilis [(90.41±7.33)% and (96.02±6.14)% of inhibition]; Avicennia germinans and Conocarpus erectus had activity against Escherichia coli [(71.17±6.15)% and (60.60±5.13)% of inhibition, respectively]. About 60% of the mangroves showed α-glucosidase inhibitory activity. In particular, extracts from Laguncularia racemosa, Pelliciera rhizophorae, Conocarpus erectus, Mora oleifera, and Tabebuia palustris species showed α-glucosidase inhibitory potential, with ICConclusions:Panamanian mangroves are mainly a promising potential source of hypoglycemic compounds, specifically α-glucosidase inhibitors. These results highlight the therapeutic virtues of extracts from American mangrove plants.

Evaluation of antiparasitic, anticancer, antimicrobial and hypoglycemic properties of organic extracts from Panamanian mangrove plants

Objective: To investigate 33 organic extracts of mangrove plants for: antiparasitic, anticancer, and antibacterial activities, as well as their ability to inhibit the activity of the -glucosidase enzyme. Methods: Leaves from all different plant mangrove species located in five mangrove zones of the Pacific coast of Panama were collected according to standard procedures. Qualitative phytochemical analysis of the organic extracts was performed by thin layer chromatography. The antiparasitic activity against Plasmodium falciparum, Trypanosoma cruzi and Leishmania donovani, toxicity against Artemia salina, anticancer activity in MCF-7 cell line, and antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa of all organic extract were investigated according protocols stablished in our institution. Finally, the ability to inhibit the enzymatic activity of -glucosidase was evaluated by monitoring the hydrolysis of p-nitrophenyl -Dglucopyranoside. Results: Thirty-three different samples belonging to nine different species of vascular plants with seeds of true mangroves were collected. Triterpenoids, phenolics, and tannins were the main groups of compounds found in the sampled mangroves. Saponins, quinones, and coumarins were found in less than 50% of the samples. Laguncularia racemosa showed moderate activity against Plasmodium falciparum. None of the extracts presented anticancer activity. Rhizophora mangle exhibited potent activity against Staphylococcus aureus and Bacillus subtilis [(90.41±7.33)% and (96.02±6.14)% of inhibition]; Avicennia germinans and Conocarpus erectus had activity against Escherichia coli [(71.17±6.15)% and (60.60±5.13)% of inhibition, respectively]. About 60% of the mangroves showed -glucosidase inhibitory activity. In particular, extracts from Laguncularia racemosa, Pelliciera rhizophorae, Conocarpus erectus, Mora oleifera, and Tabebuia palustris species showed -glucosidase inhibitory potential, with IC50 values of (29.45±0.29), (20.60±0.70), (730.06±3.74), (25.59±0.37), and (853.39±5.30) µg/mL, respectively. Conclusions: Panamanian mangroves are mainly a promising potential source of hypoglycemic compounds, specifically -glucosidase inhibitors. These results highlight the therapeutic virtues of extracts from American mangrove plants.

Phytochemical analysis and biological activities of ethanolic extract of Curcuma longa rhizome

Abstract Curcuma longa is an important dietary plant which possess several pharmacological activities, including antioxidant, antimicrobial, anti-inflamatory, anticancer and anti clotting etc. The aim of the present study was to determine the phenolic profile of Curcuma longa and in vitro antioxidant and antidiabetic activities. In HPLC chromatogram of Curcuma longa rhizome extract 15 phenolic compounds were identified namely Digalloyl-hexoside, Caffeic acid hexoside, Curdione, Coumaric, Caffeic acid, Sinapic acid, Qurecetin-3-D-galactoside, Casuarinin, Bisdemethoxycurcumin, Curcuminol, Demethoxycurcumin, and Isorhamnetin, Valoneic acid bilactone, Curcumin, Curcumin-O-glucuronide respectively. The ethanolic extract displayed an IC50 value of 37.1±0.3 µg/ml against alpha glucosidase. The IC50 value of DPPH radical scavenging activity was 27.2 ± 1.1 g/mL. It is concluded that ethanolic extract of Curcuma long is rich source of curcumin and contain several important phenolics. The in vitro antioxidant and alpha glucosidase inhibitory effect of the plant justifies its popular use in traditional medicine.

Resumo A Curcuma longa é uma importante planta presente na dieta da população, pois possui diversas atividades farmacológicas, incluindo antioxidante, antimicrobiana, anti-inflamatória, anticancerígena, anticoagulante etc. O objetivo do presente estudo foi elucidar o perfil fenólico da Curcuma longa e determinar as atividades antioxidante e antidiabética in vitro do extrato. No cromatograma por HPLC do extrato de rizoma de Curcuma longa, foram identificados 15 compostos fenólicos: digaloil-hexosídeo, hexosídeo de ácido cafeico, curdiona, cumárico, ácido cafeico, ácido sinápico, quercetina-3-D-galactosídeo, casuarinina, bisdemetoxicurcumina, curcuminol, demetoxicurcumina, isoramnetina, bilactona de ácido valônico, curcumina e curcumina-O-glicuronídeo. O extrato etanólico apresentou um valor de IC50 de 37,1 ± 0,3 µg / mL em relação à alfa-glucosidase. O valor de IC50 da atividade de eliminação de radicais DPPH foi de 27,2 ± 1,1 g / mL. Conclui-se que o extrato etanólico de Curcuma longa é uma rica fonte de curcumina e contém várias substâncias fenólicas importantes. O efeito antioxidante in vitro e inibidor da alfa-glucosidase da planta justifica seu uso popular na medicina tradicional.

Antioxidant, anti-alpha-glucosidase and pancreatic beta-cell protective effects of methanolic extract of Ensete superbum Cheesm seeds

Objective: To investigate the antioxidant, anti-a-glucosidase and pancreatic b-cell protective potential of Ensete superbum (E. superbum) seeds. Methods: A variety of in vitro assays including radical scavenging, reducing power potential, phenolic content determination, a-glucosidase assay and pancreatic b-cell (1.4E7 cells) viability were employed for assessing the effect of methanolic extract of E. superbum seeds. Results: The radical scavenging and reducing power effects comparable with the stan-dard rutin were obtained while the enzyme inhibitory activity of the extract was 68-fold better than the standard antidiabetic drug, acarbose. The seed extract of E. superbum was packed-full of polyphenols with mean percentage gallic acid equivalent value of (38.2 ± 1.8) (n = 3). The protection of pancreatic cells from massive onslaught of hydrogen peroxide was far superior to that obtained for rutin. Conclusions: The reputed antidiabetic therapeutic uses of the seeds extract of E. superbum may be justified on the basis of inhibition of carbohydrate enzymes, anti-oxidant effects and pancreatic b-cell protection.

Antioxidant, anti-alpha-glucosidase and pancreatic beta-cell protective effects of methanolic extract of Ensete superbum Cheesm seeds

Objective To investigate the antioxidant, anti-α-glucosidase and pancreatic β-cell protective potential of Ensete superbum (E. superbum) seeds. Methods A variety of in vitro assays including radical scavenging, reducing power potential, phenolic content determination, α-glucosidase assay and pancreatic β-cell (1.4E7 cells) viability were employed for assessing the effect of methanolic extract of E. superbum seeds. Results The radical scavenging and reducing power effects comparable with the standard rutin were obtained while the enzyme inhibitory activity of the extract was 68-fold better than the standard antidiabetic drug, acarbose. The seed extract of E. superbum was packed-full of polyphenols with mean percentage gallic acid equivalent value of (38.2 ± 1.8) (n = 3). The protection of pancreatic cells from massive onslaught of hydrogen peroxide was far superior to that obtained for rutin. Conclusions The reputed antidiabetic therapeutic uses of the seeds extract of E. superbum may be justified on the basis of inhibition of carbohydrate enzymes, antioxidant effects and pancreatic β-cell protection.

Antioxidant, cytotoxic and alpha-glucosidase inhibition activities from the Mexican berry "Anacahuita" (Cordia boissieri)

This study describes the total phenolic and flavonoid content as well as cytotoxic, alpha-glucosidase inhibition and antiradical/antioxidant potential of extracts obtained from the edible fruits of Cordia boissieri, which is widely distributed throughout northeastern Mexico. Phenolic and flavonoid content were evaluated by means of the Folin-Ciocalteu method and aluminum chloride colorimetric assay respectively. The antiradical/antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and Trolox Equivalent Antioxidant Capacity (TEAC) assays. Cytotoxic activity was assessed by means of human cancer cell lines (MCF-7 and HeLa), alpha-glucosidase inhibition was determined by colorimetric assay using p-Nitrophenyl α-D-glucopyranoside (PNPG) as a substrate. Results indicate that extract of C. boissieri fruit has a good antioxidant potential to show a EC50: 137.76 ± 35 μg/mL and 65 ± 2 μM/g in the DPPH and TEAC assays respectively, inhibitor of the enzyme alpha-glucosidase involved in sugar uptake (IC50: 215.20 ± 35 μg/mL), cytotoxic activities against MCF-7 (IC50: 310 ± 42 μg/mL) and HeLa (IC50: 450.4 ± 21 μg/mL) cancer cell lines as well as an important phenolic content with 230 ± 23 mg/100g and 54± 11 mg/100g of phenols and flavonoids totals respectively. These results point towards an interesting potential for the fruits of C. boissieri as chemopreventive properties and expand the possibilities for agro-industrial uses(AU)

Este estudio describe el contenido de fenoles y flavonoides totales, el efecto citotóxico, la inhibición de la enzima alfaglucosidasa y el potencial antirradical/ antioxidante del extracto obtenidos a partir de los frutos de Cordia boissieri, especie distribuida por todo el noreste de México. El contenido de fenoles y flavonoides totales se determinó por medio de los métodos de Folin-Ciocalteu y cloruro férrico respectivamente. La actividad antirradical / antioxidante se determinó mediante el secuestro del radical libre 2,2-difenil-1-picrilhidrazil (DPPH) y el ensayo de Capacidad Antioxidante Equivalente al Trolox (CAET). La actividad citotóxica se evaluó sobre las líneas celulares de cáncer humano (MCF-7 y HeLa), para determinar la inhibición de la enzima alfa-glucosidasa se utilizó el ensayo colorimétrico utilizando como sustrato p-Nitrofenil-α-D-Glucopiranósido (PNPG). Los resultados indican que el extracto del fruto de C. boissieri tiene un buen contenido de antioxidantes al mostrar una CE50 de 137.76 ± 35 μg/mL y de 65 ± 2 μM/g en los ensayos de DPPH y CAET respectivamente, un efecto inhibitorio interesante sobre la enzima alfa-glucosidasa, implicadas en la absorción de azúcar (CI50: 215.20 ± 35 μg/mL), efecto citotóxico contra las células cancerosas MCF-7 (CI50: 310 ± 42 μg/mL) y HeLa (450.4 ± 21 μg/mL), así como un importante contenido compuestos fenólicos con 230 ± 23 mg / 100g y 54 ± 11 mg / 100 g de fenoles y flavonoides totales, respectivamente. Estos resultados sugieren el potencial del fruto de C. boissieri como una fuente importante de compuestos quimiopreventivos y amplían las posibilidades para su aprovechamiento agroindustrial(AU)

Diagnóstico y tratamiento de la enfermedad de Pompe / Diagnosis and treatment of Pompe disease

INTRODUCCIÓN: la enfermedad de Pompe es una glucogenosis tipo II con patrón de herencia autosómico recesivo, causada por la deficiencia de la enzima alfa-glucosidasa ácida, que provoca acúmulo de glucógeno en los lisosomas de los macrófagos. El gen está mapeado en el cromosoma 17q25, y han sido detectadas más de 460 mutaciones. Están descritas 3 formas clínicas de la enfermedad, que incluyen, la variedad infantil, la juvenil y la tardía. La variedad infantil se caracteriza por miocardiopatía hipertrófica o infiltrativa, hipotonía, hepatomegalia y macroglosia. En la variedad juvenil hay fiebre recurrente, infecciones respiratorias a repetición y discapacidad motora progresiva. La terapia de reemplazo enzimático modifica el curso clínico de la enfermedad. PRESENTACIÓN DE CASOS: se presentan los elementos al diagnóstico de 4 casos con esta enfermedad, 3 de ellos con la variedad infantil y uno con la variedad juvenil, y se discuten los resultados de estudio molecular. Se informa la evolución clínica de 2 de ellos, luego de 6 meses y un año de tratamiento con la terapia de reemplazo enzimático con alfa-glucosidasa ácida (myozyme), lo que ha llevado a la mejoría de parámetros bioquímicos y clínicos en ambos casos. CONCLUSIONES: en pacientes con enfermedad de Pompe la cuidadosa evaluación clínica para diagnosticar precozmente la enfermedad y el comienzo de la terapia de reemplazo enzimático contribuyen a mejorar las funciones musculares, motoras y cardiovasculares, lo cual impacta en la supervivencia y calidad de vida de estos niños(AU)

INTRODUCTION: Pompe disease is a type II glycogenosis with recessive autosomal inheritance pattern, which is caused by the acid alpha-glucosidase enzyme that brings about buildup of glycogen in macrophage lysosomes. The gene is already mapped in Chromosome 17q25 and has been so far detected over 460 mutations. Three clinical forms of disease were described, including infantile, juvenile and late-onset varieties. The infantile variety is characterized by hpertrophic or infiltratin myocardiopathy, hypotonia, hepatomegaly and macroglossia. The juvenile type comprises recurrent fever, repeated respiratory infections and progressive motor disability. Enzyme replacement therapy changes the clinical course of disease. CASE PRESENTATION: the elements for diagnosis of 4 cases were submitted, 3 of them with infantile form and one with juvenile one. The results of the molecular study were discussed. The clinical progress of 2 of them after 6 monts and one year of treatment with the enzymatic replacement therapy using acid alpha-glucosidase (myozyme), which led to improvement of biochemical and clinical parameters in both cases. CONCLUSIONS: careful clinical assessment of patients with Pompe disease to early diagnose it and the onset of the enzyme replacement therapy help to improve muscle, motor and cardiovascular functions, which has an impact on survival and quality of life of these children(AU)

Guidelines for the diagnosis, treatment and clinical monitoring of patients with juvenile and adult Pompe disease / Diretriz para o diagnóstico, tratamento e acompanhamento clínico de pacientes com doença de Pompe juvenil e do adulto

ABSTRACT Pompe disease (PD) is a potentially lethal illness involving irreversible muscle damage resulting from glycogen storage in muscle fiber and activation of autophagic pathways. A promising therapeutic perspective for PD is enzyme replacement therapy (ERT) with the human recombinant enzyme acid alpha-glucosidase (Myozyme®). The need to organize a diagnostic flowchart, systematize clinical follow-up, and establish new therapeutic recommendations has become vital, as ERT ensures greater patient longevity. A task force of experienced clinicians outlined a protocol for diagnosis, monitoring, treatment, genetic counseling, and rehabilitation for PD patients. The study was conducted under the coordination of REBREPOM, the Brazilian Network for Studies of PD. The meeting of these experts took place in October 2013, at L’Hotel Port Bay in São Paulo, Brazil. In August 2014, the text was reassessed and updated. Given the rarity of PD and limited high-impact publications, experts submitted their views.

RESUMO A doença de Pompe (DP) é uma doença grave, potencialmente letal, devida ao depósito de glicogênio na fibra muscular e ativação de vias autofágicas. Tratamento promissor para a DP é a reposição enzimática com a enzima recombinante humana alfa-glicosidase ácida (rhAGA -Myozyme®). A necessidade de organizar uma propedêutica diagnóstica, sistematizar o seguimento clínico e sedimentar as novas recomendações terapêuticas tornaram-se vitais à medida que o tratamento permite uma maior longevidade aos pacientes. Uma força-tarefa de clínicos experientes no manejo da DP foi constituída para elaborar um protocolo para o diagnóstico, acompanhamento clínico, tratamento, aconselhamento genético, entre outras considerações voltadas ao paciente adulto. O estudo foi realizado sob a coordenação da Rede Brasileira de Estudos da Doença de Pompe (REBREPOM). Diante da raridade da DP e escassez de trabalhos de alto impacto de evidência científica, os especialistas emitiram suas opiniões.

Analysis on novel mutations in GAA gene of a Chinese family with two siblings affected with juvenile onset form glycogen storage disease Ⅱ / 重庆医学

Objective To identify a novel pathogenicity mutation of acid alpha‐glucosidase(GAA) gene in a Chinese family with two siblings affected with juvenile onset form glycogen storage disease Ⅱ(GSD Ⅱ) .Methods The clinical and family data of two siblings presenting recurrent respiratory tract infections ,respiratory failure associated with systemic muscle weakness ,were an‐alyzed and diagnosed with GSD Ⅱ by detecting alpha‐1 ,4‐glucosidase activity .DNA was extracted from peripheral blood of the proband ,younger brother and his parents .All 20 exons and the intron‐exon splice sites of GAA gene were amplified by polymerase chain reaction (PCR) .Mutations were detected by direct sequencing the PCR products .Results The younger brother was found to be compound heterozygous for two mutations in the GAA gene :c .1216G>A (p .Asp406Asn) missense mutation in the exon 8 from his father and c .1935C>A (p .Asp645Glu) missense mutation in the exon 14 from his mother .Conclusion The compound hetero‐zygous c .1216G>A and c .1935C>A mutations caused the juvenile onset form GSD Ⅱ characterized by dyspnea and cardiac hyper‐trophy .The novel c .1216G>A mutation may be related to the juvenile onset form GSD Ⅱ .

Postprandial hypoglycemic effects of mulberry twig and root bark in vivo and in vitro / 한국영양학회지

PURPOSE: Our previous study demonstrated the hypoglycemic effects of mulberry (Morus alba L.) leaf and the underlying mechanisms. Here we explored the potency of mulberry twigs (TW) and root barks (RB) in postprandial hypoglycemic effects in vitro and in vivo. METHODS: The major components of TW and RB were determined by high performance liquid chromatography (HPLC). Alpha-glucosidase inhibition and glucose/fructose uptake inhibition in Caco-2 cells were determined for TW, RB, and their major components, followed by an oral sugar tolerance test (OSTT) in streptozotocin-induced diabetic rats. Male Wistar rats were fed a high-fat diet for 2 weeks and then a single dose of streptozotocin (35 mg/kg B.W) was administered by intraperitoneal injection. Rats with fasting blood glucose levels above 126 mg/dL were randomly divided into 5 groups (n = 8/group) for the following treatments by gavage for 4 weeks: vehicle (normal control and diabetic control), 200 mg/kg B.W of TW or RB or 100 mg/kg B.W of oxyresveratrol (OXY). RESULTS: OXY and mulberroside A were identified as the major components of TW and OXY, mongolicin, and kuwanon H for RB. A significant inhibitory activity on alpha-glucosidase was found for TW, RB, and OXY (p = 0.0099). There was a dose-dependent inhibition of TW and RB on the intestinal sugar uptakes in Caco-2 cells, showing a greater impact on fructose compared to glucose. The OSTT showed that TW and RB significantly delayed time to maximal concentration (p = 0.0088) and decreased maximal concentration (p = 0.0043) compared to the control group. CONCLUSION: These results suggest that TW and RB may have a postprandial hypoglycemic effect, particularly in the case of high fructose or sucrose intake. OXY was suggested as a contributor to the hypoglycemic effect of TW and RB. Further studies are needed for the systemic effect of TW and RB in circulation.

Caso familiar com diagnóstico para doença de Fabry / Familiar case with diagnostics for Fabry disease

A doença de Fabry é uma enfermidade genética ligada ao cromossomoX e de caráter progressivo, causada pela deficiênciaparcial ou total da enzima alfa galactosidase A (?-Gal A). Habitualmenteo diagnóstico é tardio em função das complicaçõespatológicas provocadas pela deficiência da enzima. OBJETIVO:Neste estudo, descrevemos os aspectos clínicos de um caso familiaratravés do acompanhamento ao longo de 3 anos, duranteo tratamentopela reposição enzimática. MÉTODOS: O métodoadotado foi indutivo, relacionado ao estudo de caso familiarde pacientes com doença de Fabry. Quanto à natureza das informações,a pesquisafoi qualitativa, utilizando-se, quanto aoseu objetivo, à pesquisa exploratória. Com relação as fonte deinformação e procedimento de coleta, a pesquisa caracteriza-secomo sendo bibliográfica e documental. A amostra foi compostapor três pacientesque realizamacompanhamento quinzenalpara aplicação de terapia de reposição enzimática. O critério deinclusão para a pesquisa partiu do pressuposto de se considerarque a doença de Fabry é uma afecção rara e que a família estudadacontempla com riqueza manifestações clínicas, capazesde caracterizar a doença de Fabry. RESULTADOS: Os principaissintomas clínicos relatados pelos pacientes foram: crisede dor generalizada, fadiga, acroparestesia, febre, mialgia, dorabdominal, hipohidrose, intolerância ao frio, calor e ao exercíciofísico. Esses sintomas segundo os pacientes surgiram nainfância e foram amenizados após o uso da terapia de reposiçãoenzimática, propiciando uma melhor qualidade de vida para osmesmos. Também, se observou sinais específicos desta patologianos pacientes, como córnea verticillata e angioqueratoma. Atravésda genotipagem se verificou a semelhança da mutação entre os pacientes do estudo, demonstrando padrão típico de herançarecessiva ligada ao cromossomo X. CONCLUSÃO: Os pacientesdeste estudo apresentaram quadro clínico semelhante,sendoque a sintomatologia iniciou na infância. Córnea verticillata eangioqueratoma umbilical foram sinais encontrados nos pacientes do sexo masculino e são considerados manifestações clínicas frequentes desta patologia. A herança encontrada nesta amostra tem um padrão típico de herança recessiva ligada ao cromossomo X. Desta forma, apesar de ser uma afecção rara na população em geral, o diagnóstico precoce e a terapia de reposição enzimática permitem a evolução clínica favorável e a melhoria da qualidade de vida do paciente.(AU)

Fabry disease is a genetic disorder linked to the X chromosomeand progressive, caused by partial or total deficiency of alphagalactosidase A (?-Gal A). Usually the diagnosis is delayed dueto the pathological complications caused by deficiency of theenzyme. OBJECTIVE: In this study, we describe the clinicalaspects of a family case by monitoring for over three years,during the treatment by enzyme replacement. METHODS:The method adopted was inductive, related to the study of afamily case with patients with Fabry disease. About the nature ofthe information, the research was qualitative, using, as its goal,the exploratory research. Regarding the source of informationand collection procedure, the research is characterized asbibliographical and documentary. The sample was composed ofthree patients submitted to biweekly monitoring for applicationof enzyme replacement therapy. The inclusion criterion forthe research assumed to consider that Fabry disease is a raredisease and that the studied family contemplates with wealththe clinical manifestations, able to characterize the Fabrydisease. RESULTS: The main clinical symptoms related bypatients were: generalized pain crisis, fatigue, acroparesthesia,fever, myalgia, abdominal pain, hypohidrosis, intolerance tocold, heat and exercise. These symptoms according to patientsemerged in childhood and were alleviated after the use ofenzyme replacement therapy, providing a better quality of lifefor them. Also, we found specific signs of this disease in patients,as verticillata cornea and angiokeratoma. By genotyping, it wasfound the similarity of the mutation among patients in thestudy, showing typical pattern of recessive inheritance linkedto chromosome X. CONCLUSION: The patients in this study showed similar clinical condition, and the symptoms began inchildhood. Verticillata cornea and umbilical angiokeratomasigns were found in male patients and are considered commonclinical manifestations of this pathology. The heritage found inthis sample has a typical pattern of recessive inheritance linkedto chromosome X. Thus, despite being a rare disease in generalpopulation, early diagnosis and enzyme replacement therapyallow favorable clinical evolution and improved patient qualityof life.(AU)

In vitro alpha glucosidase inhibition and free-radical scavenging activity of propolis from Thai stingless bees in mangosteen orchard

ABSTRACTThe chemical component and biological activity of propolis depend on flora area of bee collection and bee species. In the study, the propolis from three stingless bee species, Lepidotrigona ventralis Smith, Lepidotrigona terminata Smith, and Tetragonula pagdeni Schwarz, was collected in the same region of mangosteen garden from Thailand. Total phenolic content, alpha glucosidase inhibitory effect, and free-radical scavenging activity using FRAP, ABTS, DPPH assays were determined. The most potent activity of propolis extract was investigated for bioactive compounds and their quantity. The ethanol extract of T. pagdeni propolis had the highest total phenolic content 12.83 ± 0.72 g of gallic acid equivalents in 100 g of the extract, and the strongest alpha glucosidase inhibitory effect with the IC50 of 70.79 ± 6.44 µg/ml. The free-radical scavenging activity evaluated by FRAP, ABTS, DPPH assays showed the FRAP value of 279.70 ± 20.55 µmol FeSO4 equivalent/g extract and the IC50 of 59.52 ± 10.76 and 122.71 ± 11.76 µg/ml, respectively. Gamma- and alpha-mangostin from T. pagdeni propolis extract were isolated and determined for the biological activity. Gamma-mangostin exhibited the strongest activity for both alpha glucosidase inhibitory effect and free-radical scavenging activity. Using HPLC quantitative analysis method, the content of gamma- and alpha-mangostin in the extract was found to be 0.94 ± 0.01 and 2.77 ± 0.08% (w/w), respectively. These findings suggested that T. pagdeni propolis may be used as a more suitable raw material for nutraceutical and pharmaceutical products and these mangostin derivatives as markers.