For decades, Hibiscus sabdariffa L. and its phytochemicals have been shown to possess a wide range of pharmacologic properties. In this study, aqueous extract of Hibiscus sabdariffa (AEHS) and its bioactive constituent protocatechuic acid (PCA), have been evaluated in vitro for their antiviral activity against HSV-2 clinical isolates and anti-enzymatic activity against urease. Antiherpetic activity was evaluated by the titer reduction assay in infected Vero cells, and cytotoxicity was evaluated by the neutral red dye-uptake method. Anti-urease activity was determined by a developed Electrospray Ionization-Mass Spectrometry (ESI-MS)-based assay. PCA showed potent anti-HSV-2 activity compared with that of acyclovir, with EC50 values of 0.92 and 1.43 µg∙mL(-1), respectively, and selectivity indices > 217 and > 140, respectively. For the first time, AEHS was shown to exert anti-urease inhibition activity, with an IC50 value of 82.4 µg∙mL(-1). This, combined with its safety, could facilitate its use in practical applications as a natural urease inhibitor. Our results present Hibiscus sabdariffa L. and its bioactive compound PCA as potential therapeutic agents in the treatment of HSV-2 infection and the treatment of diseases caused by urease-producing bacteria.

• MeSH
• acyklovir farmakologie   MeSH
• antivirové látky chemie   izolace a purifikace   farmakologie   MeSH
• Cercopithecus aethiops MeSH
• Hibiscus chemie   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
• inhibiční koncentrace 50 MeSH
• inhibitory enzymů chemie   izolace a purifikace   farmakologie   MeSH
• kinetika MeSH
• lidský herpesvirus 2 účinky léků   MeSH
• polyfenoly chemie   izolace a purifikace   farmakologie   MeSH
• preklinické hodnocení léčiv MeSH
• rostlinné extrakty chemie   izolace a purifikace   farmakologie   MeSH
• ureasa antagonisté a inhibitory   chemie   MeSH
• Vero buňky MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In the current study, pyroglutamic acid (pGlu), a natural amino acid derivative, has efficiently inhibited the catalytic activities of three important enzymes, namely: Human recombinant phosphodiesterase-5A1 (PDE5A1), human angiotensin-converting enzyme (ACE), and urease. These enzymes were reported to be associated with several important clinical conditions in humans. Radioactivity-based assay, spectrophotometric-based assay, and an Electrospray Ionization-Mass Spectrometry-based method were employed to ascertain the inhibitory actions of pGlu against PDE5A1, ACE, and urease, respectively. The results unveiled that pGlu potently suppressed the activity of PDE5A1 (half-maximal inhibitory concentration; IC50 = 5.23 µM) compared with that of standard drug sildenafil citrate (IC50 = 7.14 µM). Moreover, pGlu at a concentration of 20 µg/mL was found to efficiently inhibit human ACE with 98.2% inhibition compared with that of standard captopril (99.6%; 20 µg/mL). The urease-catalyzed reaction was also remarkably inactivated by pGlu and standard acetohydroxamic acid with IC50 values of 1.8 and 3.9 µM, respectively. Remarkably, the outcome of in vitro cytotoxicity assay did not reveal any significant cytotoxic properties of pGlu against human cervical carcinoma cells and normal human fetal lung fibroblast cells. In addition to in vitro assays, molecular docking analyses were performed to corroborate the outcomes of in vitro results with predicted structure-activity relationships. In conclusion, pGlu could be presented as a natural and multifunctional agent with promising applications in the treatment of some ailments connected with the above-mentioned anti-enzymatic properties.

• MeSH
• angiotensin konvertující enzym chemie   genetika   metabolismus   MeSH
• buněčné linie MeSH
• cyklické nukleotidfosfodiesterasy, typ 5 chemie   genetika   metabolismus   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
• inhibiční koncentrace 50 MeSH
• kaptopril chemie   metabolismus   MeSH
• kyselina pyrrolidonkarboxylová chemie   metabolismus   toxicita   MeSH
• kyseliny hydroxamové antagonisté a inhibitory   metabolismus   MeSH
• lidé MeSH
• rekombinantní proteiny biosyntéza   chemie   izolace a purifikace   MeSH
• sildenafil citrát chemie   metabolismus   MeSH
• simulace molekulového dockingu MeSH
• spektrofotometrie MeSH
• terciární struktura proteinů MeSH
• ureasa antagonisté a inhibitory   metabolismus   MeSH
• vazebná místa MeSH
• viabilita buněk účinky léků   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Citrulline ureidase (CTU, EC3.5.1.20) degrades citrulline into ornithine, carbon dioxide, and ammonia. Here, we present the report on expression of recombinant CTU in Escherichia coli. The soluble and active recombinant CTU was expressed in the periplasmic space with the vector pET-22b and the His-tagged CTU was purified with Ni-Affinity Chromatography. The yield of soluble recombinant protein was significantly increased when 1% sorbitol was supplemented in medium. By using phenylisothiocyanate (PITC) pre-column derivatization HPLC, the enzyme activity of recombinant CTU was determined via measuring of the substrate citrulline and the corresponding products. Our results could be useful in the study of CTU biochemical characteristics, enzymatic preparation of ornithine and development of an enzymatic detection method of citrulline.

• MeSH
• Bacteria MeSH
• bakteriologické techniky metody   využití   MeSH
• chromatografie afinitní metody   využití   MeSH
• citrulin * izolace a purifikace   metabolismus   MeSH
• enzymatické testy metody   využití   MeSH
• Escherichia coli enzymologie   imunologie   izolace a purifikace   MeSH
• Francisella tularensis * enzymologie   izolace a purifikace   metabolismus   MeSH
• hydrolasy izolace a purifikace   MeSH
• ornithin * izolace a purifikace   metabolismus   MeSH
• rekombinantní proteiny genetika   izolace a purifikace   metabolismus   MeSH
• sorbitol diagnostické užití   MeSH
• statistika jako téma MeSH
• tularemie diagnóza   etiologie   mikrobiologie   MeSH
• ureasa izolace a purifikace   metabolismus   MeSH
• vysokoúčinná kapalinová chromatografie metody   využití   MeSH
• Publikační typ
• práce podpořená grantem MeSH

The effects of the short-term application of Ascophyllum nodosum-fermented seaweed fertilizer on the bacterial community, soil nitrogen contents, and plant growth in maize rhizosphere soil were evaluated. The changes in the bacterial community composition and nitrogen contents including those of total nitrogen (TN), nitrate nitrogen (NO3--N) and ammonium nitrogen (NH4+-N) in rhizosphere soils in response to treatment with seaweed fertilizer were determined. Furthermore, soil enzymatic activity and crop biomass were analyzed. The relative abundance of the dominant phyla varied regularly with fertilization, and bacterial α-diversity was apparently influenced by seaweed fertilizer amendment. The TN contents of all soil samples decreased gradually, and the NO3--N and NH4+-N contents of the soils treated with seaweed fertilizer were much higher than those of the control soils. Similarly, the enzymatic activities of dehydrogenase, nitrite reductase, urease, and cellulase in the soil were significantly increased on day 3, day 8, and day 13 after the application of seaweed fertilizer to the maize rhizosphere soil. However, there was no difference in the activity of soil sucrase between the treatment group and the control group. In this study, the growth of maize seedlings was confirmed to be greatly promoted by the utilization of seaweed fertilizer. These results deepen our understanding of plant-microbe interactions in agroecosystems and should benefit the wide use of seaweed fertilizer in sustainable agricultural production.

• MeSH
• Ascophyllum chemie   MeSH
• Bacteria klasifikace   MeSH
• biomasa MeSH
• dusík analýza   MeSH
• fylogeneze MeSH
• kukuřice setá růst a vývoj   mikrobiologie   MeSH
• mikrobiota * MeSH
• mořské řasy chemie   MeSH
• průmyslová hnojiva analýza   MeSH
• půda chemie   MeSH
• půdní mikrobiologie MeSH
• rhizosféra * MeSH
• vývoj rostlin MeSH
• zemědělství metody   MeSH
• Publikační typ
• časopisecké články MeSH

The variations in the soil culturable bacterial communities and biochemical parameters of early successional soils from a receding glacier in the Tanggula Mountain were investigated. We examined low organic carbon (C) and nitrogen (N) contents and enzymatic activity, correlated with fewer bacterial groups and numbers in the glacier forefield soils. The soil pH values decreased, but the soil water content, organic C and total N significantly increased, along the chronosequence. The soil C/N ratio decreased in the early development soils and increased in the late development soils and it did not correlate with the soil age since deglaciation. The activities of soil urease, sucrase, protease, polyphenol oxidase, catalase, and dehydrogenase increased along the chronosequence. The numbers of culturable bacteria in the soils increased as cultured at 25°C while decreased at 4°C from younger soils to older soils. Total numbers of culturable bacteria in the soils cultured at 25°C were significantly positively correlated to the soil total N, organic C, and soil water content, as well as the activities of soil urease, sucrase, dehydrogenase, catalase, and polyphenol oxidase. We have obtained 224 isolates from the glacier forefield soils. The isolates were clustered into 28 groups by amplified ribosomal DNA restriction analysis (ARDRA). Among them, 27 groups and 25 groups were obtained from the soils at 25°C and at 4°C incubation temperatures, respectively. These groups are affiliated with 18 genera that belong to six taxa, viz, Actinobacteria, Gammaproteobacteria, Bacteroidetes, Firmicutes, Alphaproteobacteria, and Betaproteobacteria. The dominant taxa were Actinobacteria, Gammaproteobacteria, and Bacteroidetes in all the samples. The abundance and the diversity of the genera isolated at 25°C incubation temperature were greater than that at 4°C.

• MeSH
• Bacteria klasifikace   růst a vývoj   izolace a purifikace   metabolismus   MeSH
• bakteriální nálož MeSH
• časové faktory MeSH
• DNA bakterií chemie   genetika   MeSH
• DNA fingerprinting MeSH
• dusík analýza   MeSH
• enzymy analýza   MeSH
• fylogeneze MeSH
• koncentrace vodíkových iontů MeSH
• ledový příkrov * MeSH
• molekulární sekvence - údaje MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• půda chemie   MeSH
• půdní mikrobiologie * MeSH
• sekvenční analýza DNA MeSH
• shluková analýza MeSH
• společenstvo * MeSH
• teplota MeSH
• uhlík analýza   MeSH
• voda analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Čína MeSH