High Efficiency Binding Aptamers for a Wide Range of Bacterial Sepsis Agents

( Ana Claudia Graziani ),( Maria Isabel Stets ),( Ana Luisa Kalb Lopes ),( Pedro Henrique Caires Schluga ),( Soledad Marton ),( Ieda Mendes Ferreira ),( Antero Silva Ribeiro De Andrade ),( Marco Aurel 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.4

Sepsis is a major health problem worldwide, with an extremely high rate of morbidity and mortality, partly due to delayed diagnosis during early disease. Currently, sepsis diagnosis requires bacterial culturing of blood samples over several days, whereas PCR-based molecular diagnosis methods are faster but lack sensitivity. The use of biosensors containing nucleic acid aptamers that bind targets with high affinity and specificity could accelerate sepsis diagnosis. Previously, we used the systematic evolution of ligands by exponential enrichment technique to develop the aptamers Antibac1 and Antibac2, targeting the ubiquitous bacterial peptidoglycan. Here, we show that these aptamers bind to four gram-positive and seven gram-negative bacterial sepsis agents with high binding efficiency. Thus, these aptamers could be used in combination as biological recognition elements in the development of biosensors that are an alternative to rapid bacteria detection, since they could provide culture and amplification-free tests for rapid clinical sepsis diagnosis.

Original Article : Influence of P53 on the radiotherapy response of hepatocellular carcinoma

( Ana R. Gomes ),( Ana M. Abrantes ),( Ana F. Brito ),( Mafalda Laranjo ),( Joao E. Casalta Lopes ),( Ana C. Goncalves ),( Ana B. Sarmento Ribeiro ),( Maria F. Botelho ),( Jose G. Tralhao ) 대한간학회 2015 Clinical and Molecular Hepatology(대한간학회지) Vol.21 No.3

Background/Aims: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and it has a poor prognosis and few therapeutic options. Radiotherapy is one of the most effective forms of cancer treatment, and P53 protein is one of the key molecules determining how a cell responds to radiotherapy. The aim of this study was to determine the therapeutic effi cacy of iodine-131 in three human HCC cell lines. Methods: Western blotting was used to measure P53 expression. The effects of radiotherapy with iodine-131 were assessed by using the clonogenic assay to evaluate cell survival. Flow cytometry was carried out to examine the effectsof iodine-131 on cell death, oxidative stress, reduced intracellular glutathione expression, the mitochondrial membrane potential, and the cell cycle. Results: The P53 protein was not expressed in Hep3B2.1-7 cells, was expressed at normal levels in HepG2 cells, and was overexpressed in HuH7 cells. P53 expression in the HuH7 and HepG2 cell lines increased after internal and external irradiation with iodine-131. Irradiation induced a decrease in cell survival and led to a decrease in cell viability in all of the cell lines studied, accompanied by cell death via late apoptosis/necrosis and necrosis. Irradiation with 131-iodine induced mostly cell-cycle arrest in the G0/G1 phase. Conclusions: These results suggest that P53 plays a key role in the radiotherapy response of HCC. (Clin Mol Hepatol 2015;21:257-267)

Varietal Influences on Antihyperglycemia Properties of Freshly Harvested Apples Using In Vitro Assay Models

Ana Cristina Lopes Barbosa,Marcia da Silva Pinto,Dipayan Sarkar,Chandrakant Ankolekar,Duane Greene,Kalidas Shetty 한국식품영양과학회 2010 Journal of medicinal food Vol.13 No.6

The well-known health benefits of apples have been attributed in part to the presence of polyphenols and related antioxidant capacity. The consumption of apples could provide health benefits by reducing the risk for chronic diseases such as metabolic syndrome disease, including type 2 diabetes. Thus, the objective of this study was to investigate the phenolic-linked antihyperglycemia bioactive factors in aqueous and 12% ethanol extracts of peel and pulp from 10 different freshly harvested apple varieties commonly consumed in the United States. The extracts were analyzed for total soluble phenolics, 2,2-diphenyl-1-picrylhydrazyl-linked antioxidant activity, and their associated in vitro a-glucosidase and a-amylase inhibitory activities. In general, peel extracts had higher total soluble phenolic content and related antioxidant capacity than pulp extracts. Quercetin derivatives, protocatechuic acid, chlorogenic acid, and p-coumaric acid were detected, and the amount varied significantly between aqueous and ethanolic extracts. Honeycrisp and Red Delicious varieties had the highest total phenolic contents and a significant correlation with antioxidant capacity (r¼0.91). In addition, high a-amylase and a-glucosidase inhibitory activities in aqueous pulp extracts were found. However, the peel extracts had the highest a-glucosidase inhibitory activity along with low a-amylase inhibitory activity. No correlation between a-amylase inhibitory activity and total phenolic content was observed. However, positive correlations between a-glucosidase inhibitory activity and total phenolics in aqueous (r¼0.50) and ethanolic (r¼0.70) extracts were observed. This study provides the biochemical rationale for animal and clinical studies to determine the suitable varieties with optimum bioactive factors with antihyperglycemia potential.

Alchornea glandulosa Ethyl Acetate Fraction Exhibits Antiangiogenic Activity: Preliminary Findings from In Vitro Assays Using Human Umbilical Vein Endothelial Cells

Flávia Cristine Mascia Lopes,Ana Rocha,Ana Pirraco,Luis O. Regasini,Janaina R. Siqueira,Dulce H.S. Silva,Vanderlan S. Bolzani,Iracilda Z. Carlos,Raquel Soares 한국식품영양과학회 2011 Journal of medicinal food Vol.14 No.10

Alchornea glandulosa has traditionally been used in Brazilian folk medicine for the treatment of immune-inflammatory diseases and as an antiulcer agent to heal gastric ulcer and gastritis. Angiogenesis is a complex multistep process that consists of proliferation, migration, and anastomosis of endothelial cells and has a major role in the development of pathologic conditions, such as inflammatory diseases. To investigate a possible link between the anti-inflammatory activities and antiangiogenic effects of A. glandulosa ethyl acetate fraction (AGF), this study examined which features of the angiogenic process could be disturbed by this fraction. The antiangiogenic activity of AGF was determined in vitro by using human umbilical vein endothelial cells (HUVEC), and cell viability, proliferation, apoptosis, invasion, and capillary-like structures formation were addressed. To elucidate the mechanism of action, nuclear factor κB (NFκB), a transcription factor implicated in these processes, was also evaluated in HUVEC incubated with AGF. A significant decrease in proliferation, a relevant increase in apoptosis, and a strong reduction in invasion capacity (as assessed by bromodeoxyuridine, terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling, and double-chamber assays, respectively) were observed. AGF also led to a drastic reduction in the number of capillary-like structures formed when HUVEC were cultured on growth factor–reduced Matrigel–coated plates. In addition, incubation of HUVEC with AGF resulted in reduced NFκB activity. These findings emphasize the antiangiogenic potential of AGF and support its therapeutic use for disorders that involve excessive angiogenesis, such as chronic inflammation and tumor growth.

The optimal time to perform an ophthalmic examination of patients with inflammatory bowel disease

Ana Luiza Biancardi,Leandro Lopes Troncoso,Haroldo Vieira de Moraes Jr,Cyrla Zaltman 대한장연구학회 2019 Intestinal Research Vol.17 No.1

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Cenostigma macrophyllum Tul. var. acuminata Teles Freire Fraction Leaves Stimulate Gastric Healing in Rats and Human Cell Cultures

Ana Flávia Seraine Custódio Viana,Hélio B Fernandes,Mariana H Chave,Daniel A Viana,Verlane G Santos,Ariadne C A Silva,Miriam T.P. Lopes,Rita de Cássia M Oliveira 한국식품영양과학회 2021 Journal of medicinal food Vol.24 No.3

Cenostigma macrophyllum Tul. var. acuminata Teles Freire (Leguminosae–Caesalpinioideae) is a medicinal plant traditionally used for treatment of gastric ulcer. This study evaluated the ulcer-healing activity of the hydroalcoholic fraction of C. macrophyllum Tul. var. acuminata Teles Freire leaves (Cm-FHA) and the tea of the leaves of C. macrophyllum (Cm-tea), as well as the possible action of Cm-FHA, through in vitro models. Leaves of C. macrophyllum were dried and powdered to obtain the Cm-FHA. Subsequently, the Cm-FHA was characterized phytochemically and biologically. Besides, Cm-tea was prepared. The gastric healing effects of Cm-tea and Cm-FHA were analyzed using the model of acetic acid-induced gastric ulcer in rats. The human gastric adenocarcinoma (AGS) cell line was employed as an in vitro model. Cm-tea promoted a protective effect against gastric ulcers induced by absolute ethanol. Cm-FHA or Cm-tea (100 mg/kg/7 days) exhibited a significant healing effect on ulcers induced by acetic acid. In the histological analysis, gastric mucosa treated with Cm-FHA or Cm-tea advanced restoration of the mucosal epithelium. In vitro, lower concentrations of Cm-FHA stimulated cell proliferation in the BrdU assay and cell migration. Cm-tea and Cm-FHA present a significant gastric healing effect in in vivo and in vitro models.

Mechanical Properties of Alumina Nanofilled Polymeric Composites Cured with DDSA and MNA

Ana M. Amaro,Deesy G. Pinto,Luís Bernardo,Sérgio Lopes,João Rodrigues,Cristina S. Louro 한국섬유공학회 2018 Fibers and polymers Vol.19 No.2

Reinforced concrete is widely used in structures. New materials to replace both the steel and the concrete have been studied in many research centres. One of the possibilities for the reinforcement is the partial or total replacement of the steel bars by new composite materials. Nano composites are very promising, and an investigation line was developed to this end by an interdisciplinary team. On this work, the mechanical properties of epoxy resin nanocomposites (EPNCs) filled with α-Al2O3 nanoparticles (NPs) with irregular shape and approximately 100 nm maximum diameter size was investigated. The variable study was the alumina NPs contents: 1, 3 and 5 wt.%. The NPs were previously pretreated with a silane agent (APTES). Two hardeners, 3-dodec-2-enyloxolane-2,5-dione (DDSA) and 8-methyl-3a,4,7,7a-tetrahydro-4,7-methano-2- benzofuran-1,3-dione (MNA), frequently used in epoxy resin embedding tissues, were used simultaneously for this study. Unlike other hardeners, DDSA does not need curing treatment, constituting a novel application and a saving time-energy during the manufacturing process. Considering the mechanical behaviour, it was observed that the EPNCs filled with 5 wt.% of alumina NPs showed the maximum improvement in flexural modulus, around 14 % when compared to the pristine EP sample. No relevant effect was observed on the flexural strength by adding alumina NPs. Additionally, the maximum increase observed for hardness, and Young’s modulus were about 13 % and 28 %, respectively (the maximum increase was observed at 3 wt.%).

Pediatric Liver Transplantation: Caregivers' Quality of Life

Tavares, Ana Paula Bastos,Seixas, Lucas Belem Pessoa de Melo Guerra,Jayme, Caren Lopes Wanderlei,Porta, Gilda,Seixas, Renata Belem Pessoa de Melo,Carvalho, Elisa de The Korean Society of Pediatric Gastroenterology 2022 Pediatric gastroenterology, hepatology & nutrition Vol.25 No.6

Purpose: The survival rate of pediatric patients undergoing liver transplantation has increased considerably. Despite this, the period after transplantation is still complex and poses several challenges to the recipient's family, which is responsible for care management. Recently, more attention has been paid to the impact of this complex procedure on the quality of life of caregivers. Hence, this study is aimed at assessing the quality of life of caregivers of patients who have undergone liver transplantation and the aspects that influence it. Methods: This was an observational and cross-sectional study. From November 2020 to January 2021, short-form-36 questionnaires and additional questions were given to the main caregivers of children and adolescents who underwent pediatric liver transplantation. Results: Thirty-eight questionnaires were completed and the results revealed a lower quality of life in comparison to Brazilian standards, primarily in the mental domains (41.8±14.1 vs. 51.1±2.8; p<0.001). It did not show a significant association with socioeconomic or transplant-related factors, but it did show a negative impact on parents' perception of the child's health. Parents who reported worse health status for their children had a lower mental quality of life (44.1±13.8 vs. 33.3±12.6; p<0.05). Conclusion: The caregivers of transplanted children have a lower quality of life than those of the local population. Psychological assistance should be routinely provided to parents for long-term follow-up to mitigate potential negative effects on the transplanted child's care.

Gastroprotective Activity of Neoglaziovia variegata (Arruda) Mez. (Bromeliaceae) in Rats and Mice

Kamila Lopes de Lira,Flávia Danniele Frota Machado,Ana Flávia Seraine Custódio Viana,Irisdalva Sousa Oliveira,Francilene Vieira da Silva,Hélio de Barros Fernandes,Jackson Roberto Guedes da Silva Almei 한국식품영양과학회 2021 Journal of medicinal food Vol.24 No.10

Neoglaziovia variegata (Arruda) Mez (Bromeliaceae) is a medicinal plant popularly known as “caroá.” The leaves are made up of highly resistant fibers, which is of great commercial value to the handicraft and textile industry. Some studies have demonstrated that ethanolic extract of N. variegata have gastroprotective properties. This study aimed to investigate the gastroprotective activity and cytoprotective mechanisms of ethyl acetate (Nv-AcOEt), hexane (Nv-Hex), and chloroform (Nv-CHCl3) fractions of N. variegata leaves. The gastroprotective activity of Nv-AcOEt, Nv-Hex, and Nv-CHCl3 was evaluated using the ethanol and ethanol/HCl-induced gastric injury model. To elucidate the gastroprotective mechanisms, the functions of prostaglandins (PGs), nitric oxide (NO), and KATP channels were evaluated. In addition, the nonprotein sulfhydryl groups and the mucus content in the gastric tissues were analyzed. All fractions of N. variegata leaves at oral doses of 100, 200, and 400 mg/kg significantly decreased ethanol and ethanol/HCl-induced gastric lesions, leading to gastroprotection, accompanied by an increase in reduced glutathione (GSH) and gastric mucus. Gastroprotective activity of Nv-AcOEt was inhibited after pretreatment with ibuprofen and N(G)-nitro-L-arginine (L-NOARG). Gastroprotective effect of Nv-Hex and Nv-CHCl3 was also inhibited after pretreatment with L-NOARG and with glibenclamide. The results indicate that N. variegata (Arruda) Mez exhibits promising gastroprotective activity with the possible participation of NO, PGs, mucus, sulfhydryl groups, and KATP.

In Vitro Blood Flow and Cell-Free Layer in Hyperbolic Microchannels: Visualizations and Measurements

Raquel O. Rodrigues,Raquel Lopes,Diana Pinho,Ana I. Pereira,Valdemar Garcia,Stefan Gassmann,Patrícia C. Sousa,Rui Lima 한국바이오칩학회 2016 BioChip Journal Vol.10 No.1

Red blood cells (RBCs) in microchannels has tendency to undergo axial migration due to the parabolic velocity profile, which results in a high shear stress around wall that forces the RBC to move towards the centre induced by the tank treading motion of the RBC membrane. As a result there is a formation of a cell free layer (CFL) with extremely low concentration of cells. Based on this phenomenon, several works have proposed microfluidic designs to separate the suspending physiological fluid from whole in vitro blood. This study aims to characterize the CFL in hyperbolic-shaped microchannels to separate RBCs from plasma. For this purpose, we have investigated the effect of hyperbolic contractions on the CFL by using not only different Hencky strains but also varying the series of contractions. The results show that the hyperbolic contractions with a Hencky strain of 3 and higher, substantially increase the CFL downstream of the contraction region in contrast with the microchannels with a Hencky strain of 2, where the effect is insignificant. Although, the highest CFL thickness occur at microchannels with a Hencky strain of 3.6 and 4.2 the experiments have also shown that cells blockage are more likely to occur at this kind of microchannels. Hence, the most appropriate hyperbolic-shaped microchannels to separate RBCs from plasma is the one with a Hencky strain of 3. Red blood cells (RBCs) in microchannels has tendency to undergo axial migration due to the parabolic velocity profile, which results in a high shear stress around wall that forces the RBC to move towards the centre induced by the tank treading motion of the RBC membrane. As a result there is a formation of a cell free layer (CFL) with extremely low concentration of cells. Based on this phenomenon, several works have proposed microfluidic designs to separate the suspending physiological fluid from whole in vitro blood. This study aims to characterize the CFL in hyperbolic-shaped microchannels to separate RBCs from plasma. For this purpose, we have investigated the effect of hyperbolic contractions on the CFL by using not only different Hencky strains but also varying the series of contractions. The results show that the hyperbolic contractions with a Hencky strain of 3 and higher, substantially increase the CFL downstream of the contraction region in contrast with the microchannels with a Hencky strain of 2, where the effect is insignificant. Although, the highest CFL thickness occur at microchannels with a Hencky strain of 3.6 and 4.2 the experiments have also shown that cells blockage are more likely to occur at this kind of microchannels.Hence, the most appropriate hyperbolic-shaped microchannels to separate RBCs from plasma is the one with a Hencky strain of 3.