Antibacterial and hemolytic activity of extracts and compounds obtained from epicarps and seeds of Garcinia madruno(Kunth) Hammel / Actividad antimicrobiana y hemolítica de extractos y compuestos obtenidos del epicarpio y las semillas de Garcinia madruno (Kunth) Hammel

This paper describes the evaluation of the antimicrobial and hemolytic activity of the hexane, dichloromethane, ethyl acetate and methanol extracts from seeds and epicarps of Garcinia madruno; as well garcinol, morelloflavone and volkensiflavone isolated from the same species. In the preliminary test of bacterial susceptibility, hexane extracts from seeds and epicarps and the three compounds tested only displayed inhibitory growth effect against Gram-positive bacteria. The minimum inhibitory concentrations of extract and compounds ranging from 86.6 to 1253.4 µg/mL. The hemolytic activity was assessed; however, except for the methanol extract from seeds, none of the samples studied induced hemolysis. Thus, our results suggest that extracts and compounds from G. madruno have the potential to be used in the control of pathologies associated to Gram-positive bacteria. This is the first report of the antimicrobial and hemolytic activity of extracts of different polarity obtained from seeds and epicarps of this edible species.

El presente artículo describe la evaluación de la actividad antimicrobiana y hemolítica de los extractos de hexano, diclorometano, acetato de etilo y metanol, obtenidos de la semilla y el epicarpio de Garcinia madruno; así como de garcinol, morelloflavona y volkensiflavona; aislados de la misma especie. En el ensayo de susceptibilidad bacteriana, tanto el extracto de hexano obtenido a partir de la semilla y el epicarpio, y los tres compuestos aislados, únicamente mostraron actividad inhibitoria del crecimiento contra bacterias Gram-positivas. La concentración mínima inhibitoria presentó valores entre 86.6 y 1253.4 µg/mL. También se estableció la actividad hemolítica; sin embargo, con excepción del extracto metanólico obtenido a partir de las semillas, ninguna de las muestras evaluadas indujo hemólisis. Por lo tanto, los resultados sugieren que los extractos y compuestos de G. madruno tienen el potencial de ser usados en el control de bacterias Gram-positivas asociadas a diversas patologías. Este es el primer reporte de actividad antimicrobiana y hemolítica de extractos de diferente polaridad obtenidos de las semillas y epicarpios de esta especie comestible.

Molecular identification of microbial contaminations in the fitness center in Makkah region

As the condition increases and seeks to remain healthy, the number of people who plan to join a fitness center or "gym" has increased markedly. From where this individual does understand, the study of the variety of bacteria showing the stolen has led him to care for people, with a popular fitness center located in the province of Makka, Saudi Arabia. Different bacteria must be eliminated from other gyms in Makkah, Saudi Arabia, in a total of three areas 46 two sports equipment. Both types and characteristics of bacteria, while some have been tested in hemolytic surgery for antibiotic resistance. Corynebacterium antibiotics in different forms did not react the same; however, isolates tested for M17 and N12 showed the greatest resistance to antibiotics. Furthermore, sixteen bacterial strains of human blood ß-agar displayed high hemolytic activity. In the gym isolates 2 (9 strains) followed in gym 1 (7 modes), B row hemolytic activity was highest. It is important to note that gram-positive bacteria were positive in all kinds of ways, and catalase was positive. Six strains belonging to the genus Bacillus, Brachybacterium, Geobacillus, Microbacterium, Micrococcus, and Staphylococcus and other pathogenic bacteria were known as possible individuals to use the morphological, biochemical, and rRNA gene of the 16S series. In general, this research illustrates the health and fitness centers in the individual being studied and the risks that are considered necessary to periodically study possible microbial contamination in the mixture in the gym to ensure people's protection.

Structure modification and antimicrobial activity of novel cationic melittin analogues / 药学学报

Melittin exhibits high antibacterial potency against drug-resistant bacteria. However, the clinical utility of melittin is limited by its serious hemolytic activity. Thus, the need for developing novel melittin analogues with high antimicrobial activity and low hemolytic activity has grown. We designed, synthesized, and evaluated 20 novel melittin analogues with varying hydrophobic, polar or positively charged amino acids. The results showed that 8 compounds had antimicrobial activity (MIC: 1-4 μg·mL-1) against gram-positive pathogens equal to or better than that of melittin, and 16 compounds had low hemolytic activity (HC50 ≥ 11.9 μg·mL-1). Compounds 13 (MIC: 2-4 μg·mL-1) and 15 (MIC: 1-2 μg·mL-1) showed equal or better antimicrobial activity against both susceptible and resistant strains of Staphylococcus aureus and Enterococcus faecium compared to melittin (MIC: 4 μg·mL-1). Compound 13 (HC50: 24.0 ± 4.3 μg·mL-1) displayed noticeably decreased hemolytic activity compared to melittin (HC50: 5.3 ± 0.4 μg·mL-1). This work established a base for further study on the structure-activity relationships and structure-toxicity relationships of melittin.

The essential oil of Curcuma longa rhizomes as an antimicrobial and its composition by Gas Chromatography/Mass Spectrometry / Óleo essencial de Curcuma longa L. como um antimicrobiano e composição por Cromatografi a Gasosa/Espectrometria de massas

Objective This study aimed to extract the essential oil of Curcuma longa rhizomes collected in Brazil, determine its composition by gas chromatography and mass spectrometry, and evaluate its hemolytic action and antimicrobial activity. Methods The oil extraction was performed by hydrodistillation; its composition was determined by GC-MS; the Minimum Inhibitory Concentration was evaluated through microdilution, and the hemolytic activity was analyzed in sheep red blood cells. Results The essential oil's major components are zingiberene (11%), sesquiphellandrene (10%), ß-turmerone (10%), and α-curcumene (5%). It proved efficient at inhibiting Staphylococcus aureus with Minimum Inhibitory Concentrations of 38.8µl/mL, Staphylococcus epidermidis (Minimum Inhibitory Concentrations of 50.0µl/mL), Escherichia coli (Minimum Inhibitory Concentrations of 44.4µl/mL), and Pseudomonas aeruginosa (Minimum Inhibitory Concentrations of 27.7µl/mL). Conclusion Despite provoking hemolysis in sheep red blood cells, the essential oil suggests promising results for a variety of purposes due to its antibacterial properties. Supplementary research is necessary to determine in vivo activity and the potential use of the C. longa essential oil as an antimicrobial agent in diverse situations.

Objetivo Este trabalho teve como objetivo determinar a composição e avaliar as atividades antimicrobiana e hemolítica do óleo essencial de rizomas da Curcuma longa L. Métodos A extração do óleo essencial foi realizada por hidrodestilação e sua composição determinada por cromatografia gasosa com espectrometria de massas. A Concentração Mínima Inibitória foi avaliada por microdiluição e a atividade hemolítica foi analisada em hemácias de carneiro. Resultados Os componentes majoritários encontrados foram zingibereno (11%), sesquipelenadieno (10%), ß-turmerona (10%) e α-curcumeno (5%). O óleo essencial provocou hemólise e inibiu Staphylococcus aureus (Concentração Mínima Inibitória=38,8µl/mL), Staphylococcus epidermidis (Concentração Mínima Inibitória=50,0µl/mL), Escherichia coli (Concentração Mínima Inibitória=44,4µl µl/mL) e Pseudomonas aeruginosa (Concentração Mínima Inibitória=27,7µl/mL). Conclusão Apesar de provocar hemólise, o óleo essencial analisado tem potencial para diversas finalidades, devido às suas propriedades antibacterianas. Estudos complementares são necessários para determinar a atividade in vivo e o potencial uso de óleo essencial de C. longa como agente antimicrobiano em diversas situações.

Novel Cathelicidin Antimicrobial Peptides from Paa robertingeri

This study aimed to describe two cathelicidins (cathelicidin-PR1 and cathelicidin-PR2) from the skin of Paa robertingeri (Anura: Ranidae). The deduced mature peptides cathelicidin-PR1 and cathelicidin-PR2 were composed of 29 and 25 residues, respectively. Cathelicidin - PR1 has higher antimicrobial activity it could kill Gram-positive and Gram-negative bacteria and even some fungal species. Cathelicidin-PR1 exhibited more effective than AMP in antimicrobial activity against Pseydomonas maltophilia clinical strain. On the contrary, cathelicidin-PR2 had very weak antimicrobial activity. Furthermore, cathelicidin-PR1 and cathelicidin-PR2 exhibited very low hemolytic activity against human erythrocytes and little hemagglutinating activity. The results suggested that the cathelicidin-PR1 might serve as a template for developing novel antibiotics.

Synthesis and in silico study of 2-furyl(4-{4-[(substituted)sulfonyl]benzyl}-1-piperazinyl)methanone derivatives as suitable therapeutic agents

Abstract In the study presented here, a new series of 2-furyl(4-{4-[(substituted)sulfonyl]benzyl}-1-piperazinyl)methanone derivatives was targeted. The synthesis was initiated by the treatment of different secondary amines (1a-h) with 4-bromomethylbenzenesulfonyl chloride (2) to obtain various 1-{[4-(bromomethyl)phenyl]sulfonyl}amines (3a-h). 2-Furyl(1-piperazinyl)methanone (2-furoyl-1-piperazine; 4) was then dissolved in acetonitrile, with the addition of K2CO3, and the mixture was refluxed for activation. This activated molecule was further treated with equi-molar amounts of 3a-h to form targeted 2-furyl(4-{4-[(substituted)sulfonyl]benzyl}-1-piperazinyl)methanone derivatives (5a-h) in the same reaction set up. The structure confirmation of all the synthesized compounds was carried out by EI-MS, IR and 1H-NMR spectral analysis. The compounds showed good enzyme inhibitory activity. Compound 5h showed excellent inhibitory effect against acetyl- and butyrylcholinesterase with respective IC50 values of 2.91±0.001 and 4.35±0.004 µM, compared to eserine, a reference standard with IC50 values of 0.04±0.0001 and 0.85±0.001 µM, respectively, against these enzymes. All synthesized molecules were active against almost all Gram-positive and Gram-negative bacterial strains tested. The cytotoxicity of the molecules was also checked to determine their utility as possible therapeutic agents.

Influence of concentrations of CaCl2 on hemolytic activity of candida strains / 口腔疾病防治

Objective@# To investigate the influence of different concentrations of CaCl2 on the hemolytic activity of candida strains. @*Methods @#A total of 25 candida strains were selected, including 21 candida strains recovered from clinical specimens, and 4 reference strains of C. albicans (ATCC 90028) (American Type Culture Collection, Manassas, VA, USA), C. glabrata (ATCC 90030), C. krusei (ATCC 6258), and C. tropicalis (ATCC 13803) respectively. CaCl2 was added at concentrations of 0%, 0.5%, 1% and 2.5 g % (wt/vol). Plates were cultured at 37 ℃ in a 5% CO2 atmosphere for 24-72 h, and the hemolytic index (Hi) was analyzed. @*Results @#comparsions of the hemolytic indices among the groups treated with 0.5% CaCl2 (Hi = 2.247 ± 0.079), 1% CaCl2 (Hi = 2.013 ± 0.092), 2.5% CaCl2 (Hi = 2.150 ± 0.066) and the control (Hi = 2.749 ± 0.103) reached statistical significance (P < 0.001). @*Conclusion@#CaCl2 may produce a decrease activity in the hemolysis of candida species.

Antioxidant and anti-inflammatory activities of Arbutus unedo aqueous extract

Objective To evaluate the antioxidant and anti-inflammatory activities of aqueous extract of Arbutus unedo (A. unedo) leaves. Methods In this context, the in vitro antioxidant activity was demonstrated by 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical and H

Antioxidant and anti-inflammatory activities of Arbutus unedo aqueous extract

Objective: To evaluate the antioxidant and anti-inflammatory activities of aqueous extract of Arbutus unedo (A. unedo) leaves. Methods: In this context, the in vitro antioxidant activity was demonstrated by 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical and H2O2 radical scavenging, ferrous ion chelating, ferric reducing power, total antioxidant capacity and by the protection against peroxidation of b-carotene-linoleic acid in emulsion. The anti-inflammatory activity was evaluated first by studying the membrane of human red blood cells against different hypotonic concentrations of NaCl and against heat, inhibiting the denaturation of albumin. Results: Total phenolic and flavonoid content were found respectively [(207.84 ± 15.03) mg gallic acid equivalent/g, and (13.070 ± 0.096) mg quercetin equivalent/g]. The extract displayed significant scavenging activity of some radicals such as 2,2-diphenyl-1-picrylhydrazyl [IC50 at (7.956 ± 0.278) mg/mL], ?OH [IC50 = (1 015.74 ± 46.35 mg/mL)], H2O2 [IC50 = (114.77 ± 16.86) mg/mL] and showed a good antioxidant activity through ferrous ion chelating activity [IC50=(1 014.30 ± 36.21) mg/mL], ferric reducing power [IC50 = (156.55 ± 17.40) mg/mL], total antioxidant capacity [IC50 = (461.67 ± 4.16) mg/mL] and b-carotene-linoleic acid protection against peroxidation [I%=(87.04 ± 1.21)%at 1 000 mg/mL]. Conclusions: A. unedo showed in vitro anti-inflammatory activity by inhibiting the heat induced albumin denaturation and red blood cells membrane stabilization. Our results show that aqueous leaf extract of A. unedo has good antioxidant activity and interesting anti-inflammatory properties. A. unedo aqueous extract can be used to prevent oxidative and inflammatory processes.

Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola / Detection of the genes encoding the toxin CDT, and research factors which influence the production of hemolysin in Campylobacter jejuni from poultry products

Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT) por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções quelantes e de íons nesta atividade. Foram utilizadas 45 amostras de Campylobacter jejuni de origem avícola para pesquisa de atividade hemolítica, cultivadas em Caldo Triptona de Soja (TSB). Após o crescimento bacteriano, as amostras foram semeadas em Ágar tríptico de soja (TSA) contendo 5% de sangue de ovino. Para verificar a influência de agentes quelantes e solução de íons na atividade hemolítica, as amostras de C. jejuni foram cultivadas em TSB contendo separadamente os quelantes EDTA, ácido acético, soluções de íons CaCl2, MgCl2 e FeCl3, em atmosfera de microaerofilia. Quanto à atividade de hemolisina de C. jejuni em placas de TSA - sangue ovino foi possível observar que houve hemólise em 40% das amostras analisadas apenas com caldo TSB. Somente o ácido acético apresentou ação quelante sobre a atividade de hemolisinas em amostras de C. jejuni semeadas em placas de TSA - sangue ovino. Para detecção dos genes cdtA, cdtB e cdtC através da técnica da Reação em Cadeia da Polimerase (PCR) foram utilizadas 119 amostras de C. jejuni de origem avícola. Foi possível observar que 37,8% possuíam o perfil de genes cdtABC. Os resultados demonstraram em amostras avícolas a presença de cepas de C. jejuni com potencial virulento, devido à presença dos genes da toxina CDT e potencial hemolítico, que apresentou ação reduzida in vitro com ácido acético...

Thermophilic members of the Campylobacter genus are recognized as important enteropathogenics for humans and animals. The great variety of ecological habitats, such as water, food and milk, may promote new virulence factors. To detect the encoding genes distending cytolethal toxin (CDT) by PCR and study the hemolytic activity with influence of chelation solutions and ions, 45 Campylobacter jejuni samples from poultry production origin were used to perform the hemolytic research. To check the influence of chelation agents and solution of ions in the hemolytic activity, samples of C. jejuni strains were grown in tryptone soy broth TSB containing chelation agents separately EDTA, acetic acid, CaCl2, MgCl2 and FeCl3 ions solutions in microaerophilic atmosphere and then streaked on 5% sheep blood tryptic soy agar (TSA). To perform the detection of cdtA, cdtB and cdtC genes the technique of Polymerase Chain Reaction (PCR) was used in 119 samples of C. jejuni from poultry production origin. We found 40% of samples showing hemolysis after growing with TSB. Only the acetic acid showed reduction in hemolysis. The prevalent gene profile was cdtABC in 37.8 % of the samples. It was observed that the results showed the presence of C. jejuni strains with virulent potential, due to presence of the CDT toxin genes and the hemolytic activity, which showed in vitro reduced when acetic acid was added...

Biological characterization of compounds from Rhinella schneideri poison that act on the complement system

Background The skin secretions of toads of the family Bufonidae contain biogenic amines, alkaloids, steroids (bufotoxins), bufodienolides (bufogenin), peptides and proteins. The poison of Rhinella schneideri, formerly classified as Bufo paracnemis, presents components that act on different biological systems, including the complement system. The aim of this study was to isolate and examine the activity ofRhinella schneideri poison (RsP) components on the complement system.Methods The components active on the complement system were purified in three chromatographic steps, using a combination of cation-exchange, anion-exchange and gel filtration chromatography. The resulting fractions were analyzed by SDS-PAGE and screened for their activity in the hemolytic assay of the classical/lectin complement pathways. Fractions active on the complement system were also assessed for their ability to generate C3 fragments evaluated by two dimensional immunoelectrophoresis assay, C3a and C5a by neutrophil chemotaxis assay and SC5b-9 complex by ELISA assay.Results The fractionation protocol was able to isolate the component S5 from theRsP, as demonstrated by SDS-PAGE and the RP-FPLC profile. S5 is a protein of about 6000 Da, while S2 presents components of higher molecular mass (40,000 to 50,000 Da). Fractions S2 and S5 attenuated the hemolytic activity of the classical/lectin pathways after preincubation with normal human serum. Both components stimulated complement-dependent neutrophil chemotaxis and the production of C3 fragments, as shown by two-dimensional immunoelectrophoresis. S2 showed a higher capacity to generate the SC5b- 9 complex than the other fractions. This action was observed after the exposure of normal human serum to the fractions.Conclusions This is the first study to examine the activity of RsP components on the complement system. Fractions S2 and S5 reduced the complement hemolytic activity, stimulated complement-dependent neutrophil chemotaxis and stimulated the production of C3 fragments, indicating that they were able to activate the complement cascade. Furthermore, fraction S2 was also able to generate the SC5b-9 complex. These components may be useful tools for studying dysfunction of the complement cascade.(AU)

Biological characterization of compounds from Rhinella schneideri poison that act on the complement system

Background The skin secretions of toads of the family Bufonidae contain biogenic amines, alkaloids, steroids (bufotoxins), bufodienolides (bufogenin), peptides and proteins. The poison of Rhinella schneideri, formerly classified as Bufo paracnemis, presents components that act on different biological systems, including the complement system. The aim of this study was to isolate and examine the activity ofRhinella schneideri poison (RsP) components on the complement system.Methods The components active on the complement system were purified in three chromatographic steps, using a combination of cation-exchange, anion-exchange and gel filtration chromatography. The resulting fractions were analyzed by SDS-PAGE and screened for their activity in the hemolytic assay of the classical/lectin complement pathways. Fractions active on the complement system were also assessed for their ability to generate C3 fragments evaluated by two dimensional immunoelectrophoresis assay, C3a and C5a by neutrophil chemotaxis assay and SC5b-9 complex by ELISA assay.Results The fractionation protocol was able to isolate the component S5 from theRsP, as demonstrated by SDS-PAGE and the RP-FPLC profile. S5 is a protein of about 6000 Da, while S2 presents components of higher molecular mass (40,000 to 50,000 Da). Fractions S2 and S5 attenuated the hemolytic activity of the classical/lectin pathways after preincubation with normal human serum. Both components stimulated complement-dependent neutrophil chemotaxis and the production of C3 fragments, as shown by two-dimensional immunoelectrophoresis. S2 showed a higher capacity to generate the SC5b- 9 complex than the other fractions. This action was observed after the exposure of normal human serum to the fractions.Conclusions This is the first study to examine the activity of RsP components on the complement system. Fractions S2 and S5 reduced the complement hemolytic activity, stimulated complement-dependent neutrophil chemotaxis and stimulated the production of C3 fragments, indicating that they were able to activate the complement cascade. Furthermore, fraction S2 was also able to generate the SC5b-9 complex. These components may be useful tools for studying dysfunction of the complement cascade.

Study on virulence factors of Candida tropicalis isolated from clinical samples / 中华流行病学杂志

Objective To determine the in vitro production of virulence factors for Candida (C.) tropicalis,including aspartyl proteinases,phospholipases and hemolytic activities,describe the regulation of virulence factors varying with time in C.tropicalis,and analyze the differences in aspartyl proteinases and hemolytic activities of C.tropicalis isolated from anatomically distinct sites.Methods A total of 64 C.tropicalis strains were spot-inoculated onto bovine albumin agar,egg yolk agar and sheep blood agar plates,respectively.Then the plates were incubated for 24,48 and 72 hour at 37 ℃,respectively.The aspartyl proteinases,phospholipase and hemolytic activities were determined at each time point,respectively.Results All the C.tropiclais isolates showed positive aspartyl proteinases and hemolytic activities at each time point,but no phospholipases activity was detected in C.tropicalis.On comparison of aspartyl proteinases and hemolytic activities at different time points,aspartyl proteinases activity at 48 and 72 hour was higher than that at 24 hour.During 72 hour,hemolytic activity of C.tropicalis increased.No statistical significant differences in aspartyl proteinases and hemolytic activities of C.tropicalis were observed among different infection sites (P=0.368 and 0.985).Conclusion The C.tropicalis clinical isolates in China have aspartyl proteinases activity,hemolytic activity,but have no phospholipase activity.

Alleviation of PEGylated Puerarin on Erythrocyte Hemolysis Induced by Puerarin in Glucose-6-phosphate Dehydrogenase-deficient Rats / 中草药·英文版

Objective: To explore and analyze the reducing hemolytic effects of PEGylated puerarin (PEG-PUE) on erythrocytes induced by PUE in glucose-6-phosphate dehydrogenase (G6PD)-deficient rats. Methods: The rat model with G6PD-deficiency was established via sc injecting 1% acetylphenyl-hydrazine. Then the G6PD-deficient erythrocyte suspension obtained from this rat model was used to evaluate the hemolytic effects of PUE and the reducing hemolytic effects of PEG-PUE via hemolytic activity and erythrocyte osmotic fragility assay. Results: It was found that PUE could cause a serious hemolysis to the erythrocyte suspension with the increase of drug concentration and the prolongation of drug incubation time, the hemolytic rate of PUE was up to 40%, while the addition of PEG-PUE to the erythrocyte suspension revealed no significant hemolysis. Additionally, the result of erythrocyte osmotic fragility indicated that PEG-PUE exerted a slight effect on the erythrocyte membranes, and the NaCl concentration that induced 50% hemolysis (32 mmol/L) was about one-third PUE. Conclusion: These results demonstrate that PEG-PUE could play a significant role in reducing the side effect of hemolysis induced by PUE. The low hemolytic activity of PEG-PUE makes it a favorable candidate for in vivo tests and PEG-PUE could also provide the useful insight for the further formulation development as an innovative drug. © 2013 Tianjin Press of Chinese Herbal Medicines.

Protein stability and hemolytic activity of tentacle extract from the jellyfish Cya-nea capillata / 药学实践杂志

Objective To investigate the influencing factors of the protein stability and hemolytic activity of tentacle extract ( TE) from the jellyfish Cyanea capillata .Methods Effects of various factors and treatments on the protein stability and hemolytic ac-tivity of TE were explored by protein detection , hemolytic assay and SDS-PAGE analysis.Results TE caused a significant and dose-dependent hemolytic effect , and the HU50 of TE against 0.5%erythrocyte suspensions from SD rats was 226μg/ml.A 40℃water bath for 1 hour could effectively remove the contaminating proteins in TE .TE retained hemolytic activity at 4℃for 28 days but it was unsta-ble when kept at 25℃over 3 days.TE was active in the range from pH 6.0 to 11.0 and the optimum pH was 8.0.Various buffer solu-tions had significantly different effects on the stability and hemolytic activity of TE , and a good salting-out effect was observed on the hemolytic protein of TE while the concentration of ammonium sulfate solutions was greater than 26%.Conclusion A 40℃water bath for 1 hour could effectively remove the contaminating proteins in TE and reduce its viscosity .The optimum conditions for maintaining stability and hemolytic activity of TE were 4℃ and pH 8.0.The salting-out effect from 26% and more ammonium sulfate solutions would be conducive to the enrichment of hemolytic protein .

Rapid synthesis of biocompatible silver nanoparticles using aqueous extract of Rosa damascena petals and evaluation of their anticancer activity

OBJECTIVE@#To optimize the process parameters involved in the green synthesis of silver nanoparticles (G-SNPs) by aqueous extract of Rosa damascena petals and to evaluate the biocompatibility and anti cancer activity of the synthesized silver nanoparticles against human lung adenocarcinoma (A549).@*METHODS@#The process variables that include concentration of extract, mixing ratio of reactants, silver salt concentration and interaction time were analyzed. The compatibility of the G-SNPs was verified by incubating with erythrocytes and the anticancer property of the G-SNPs against A549 cells was performed by MTT assay.@*RESULTS@#Formation of G-SNPs was confirmed by the visual change in the colour of the reaction mixture from pale yellow to brown yellow. Surface plasmon resonance of synthesized G-SNPs was observed at 420 nm; the size of G-SNPs were analyzed by DLS and found to be in the range of (84.00±10.08) nm. Field emission scanning electron microscope and high resolution transmission electron microscopy analysis confirmed that the G-SNPs were fairly spherical. Fourier transform infrared spectroscopy spectroscopy and X-ray diffraction revealed the characteristic peaks of G-SNPs. Energy dispersive X-ray analysis showed a signal of silver around 3 keV. The synthesized G-SNPs exhibited anticancer activity as evidenced by the MTT assay. IC50 value of G-SNPs was found to be 80 μg/mL.@*CONCLUSION@#The results of the present study suggest that G-SNPs can be synthesized rapidly within first minute of the reaction; they are biocompatible and possess anticancer activity against human lung adenocarcinoma.

Expression, purification and antibody preparation of alpha-hemolysin from Staphylo-coccus aureus / 军事医学

Objective To clone and express alpha hemolysin ( Hla ) , one important virulence factor secreted by Staphylococcus aureus in Escherichia coli to tdetect the hemolytic activity of Hla on erythrocytes from diffrerent species ,and to prepare specific antibodies against Hla that can inhibit the hemolytic activity of Hla .Methods Hla gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as a template.The expression vector pET-28a-Hla was constructed and transformed into E.coli BL21(DE3).The recombinant Hla protein was expressed by IPTG induction ,and then purified by Ni2+affinity chromatography .The hemolytic activity of Hla was measured by erythrocyte lysis assays .Results Soluble recombinant Hla protein was expressed and purified .Further investigations showed that Hla could significantly induce the lysis of rabbit erythrocytes .However , erythrocytes from humans or sheep were more resistant to the lysis mediated by Hla . The specific polyclonal antibodies against Hla ( anti-Hla) were prepared and detected by ELISA .Moreover, it was found that anti-Hla could inhibit the hemolytic activity of Hla .Conclusion We found that Hla from S.aureus has different hemolytic effects on erythrocytes from various species .The prepared Hla-antibodies can specifically block the hemolytic activity of Hla.

Rapid synthesis of biocompatible silver nanoparticles using aqueous extract of Rosa damascena petals and evaluation of their anticancer activity

Objective: To optimize the process parameters involved in the green synthesis of silver nanoparticles (G-SNPs) by aqueous extract of Rosa damascena petals and to evaluate the biocompatibility and anti cancer activity of the synthesized silver nanoparticles against human lung adenocarcinoma (A549). Methods: The process variables that include concentration of extract, mixing ratio of reactants, silver salt concentration and interaction time were analyzed. The compatibility of the G-SNPs was verified by incubating with erythrocytes and the anticancer property of the G-SNPs against A549 cells was performed by MTT assay. Results: Formation of G-SNPs was confirmed by the visual change in the colour of the reaction mixture from pale yellow to brown yellow. Surface plasmon resonance of synthesized G-SNPs was observed at 420 nm; the size of G-SNPs were analyzed by DLS and found to be in the range of (84.00±10.08) nm. Field emission scanning electron microscope and high resolution transmission electron microscopy analysis confirmed that the G-SNPs were fairly spherical. Fourier transform infrared spectroscopy spectroscopy and X-ray diffraction revealed the characteristic peaks of G-SNPs. Energy dispersive X-ray analysis showed a signal of silver around 3 keV. The synthesized G-SNPs exhibited anticancer activity as evidenced by the MTT assay. IC

Exploración estructural de perfiles de actividad antibacteriana y hemolítica de análogos no naturales del péptido antimicrobiano Mastoparán MP-8 / Structural exploration of antibacterial activity and hemolytic profiles of non-natural analogs of the antimicrobial peptide mastoparan MP-8

En este trabajo se reporta el efecto, la modificación puntual y sistemática a nivel del enlace peptídico realizada al péptido Mastoparán MP-8, obteniéndose 27 pseudopéptidos en total. Los resultados permitieron determinar que los pseudopéptidos ψ-38617(INLKALAALAKd-[CH2NH]-RLL), ψ-38629(INLKAd-[CH2NH]-LAALAKRLL) y ψ-38630 (INLK-[CH2NH]-ALAALAKRLL) presentan mejor actividad que el péptido Mastoparán MP-8 frente a las bacterias Gram negativas y conservaron la actividad antibacteriana frente a las bacterias Gram positivas; además, la modificación realizada produjo tres moléculas que abolieron totalmente la actividad hemolítica propia del péptido nativo. Todas las modificaciones realizadas a esta secuencia provocaron un aumento en la estabilidad de las moléculas frente al ataque enzimático.

In this work the relevance of a systematic replacement of peptide-bonds on the Mastoparan MP-8 antimicrobial peptide to afford 27 new pseudopeptide analogues is reported. Results allowed to determine that pseudopeptides ψ-38617 (INLKALAALAKd-[CH2NH]-RLL), ψ-38629(INLKAd-[CH2NH]-LAALAKRLL) and ψ-38630 (INLK-[CH2NH]-ALAALAKRLL), showed an enhanced anti gam-negative bacterial properties, regarding the native Mastoparan MP-8 peptide, but maintaining a comparable activity against gram-positive bacteria. In addition, specific performed modifications on MP-8 sequence led three new molecules that completely abolished the hemolytic activity of the parent MP-8 peptide. All obtained molecules possess a high stability profile when tested against a severe proteolytic attack.