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The present study evaluates the antiarthritic effect of hydroethanolic extract of Pleurotus ostreatus cv. Florida, which was tested against adjuvant induced arthritis in rat models. Arthritis was induced by administration of complete Freund's adjuvant into the subplantar surface of left paw of rats. The extract was given orally at doses 200 mg/ kg and 400 mg/kg and piroxicam was administered intraperitonially (4 mg/kg). In vitro testing on parameters including antiproteinestrase, albumin denaturation and heat induce hemolysis was also carried out. There was significant decrease (p < 0.001) in proteinase activity and membrane stabilization in vivo studies on cv. Florida extract treated rats showed a significant (p < 0.001) decrease in paw volume, joint diameter, and spontaneous change in body weight recorded for 21 days. The treatment also resulted in an increase in rats' gripping activity compared with arthritic control rats. X-ray examinations showed a decrease in joint swelling. Histopathological examination of the extract treated group showed a significant decrease in joint space. There was also an increase in antibody levels. The antioxidant parameters showed a significant (p < 0.001) increase in superoxide dismutase and catalase enzymatic activities. Thus P. ostreatus cv. Florida extract demonstrates a potent antioxidant activity in a rat model. It is concluded that the P. ostreatus cv. Florida extract contains medicinally important constituents that show antiarthritic activity in rats.

Rotenone is a pesticide that has been shown to induce the pathological symptoms of Parkinson's disease (PD) in both cellular and animal models. In this study, we investigated the protective effect of Agaricus blazei extract on rotenone-induced dopaminergic degeneration and apoptosis in mice model. A. blazei extract blocked the rotenone-mediated diminution of   dopamine transporter (DAT)  and vesicular monoamine transporter 2 (VMAT 2) expression and  the downregulation of Bcl-2 and the upregulation of Bax, caspases-3, -6, -8 and caspase-9. Present data suggest that A. blazei extract plays a crucial role in regulation of proteins expression such as DAT and VMAT2 and pro-apoptotic and anti-apoptotic in Parkinsonism. In conclusion, the present study shows that A. blazei extract act as potential neuroprotective agent in the management of Parkinsonism.

As a natural pigment, cordycepic carotenoids have many bioactive functions, such as antiinflammation, anticancer, and antioxidation. In addition, the good coloring of this hydrophilic pigment enables it to have wide application in the food industry. This study investigated five species of fungal elicitors, namely, Rhodotorula glutinis, Saccharomyces cerevisiae, Monascus ruber, Blakeslea trispora, and Flammulina velutipes, to evaluate their effects on carotenoid accumulation in Cordyceps militaris. Results showed that all fungal elicitors, except Rh. glutinis, have no positive effect on the biosynthesis of cordycepic carotenoids. The Rh. glutinis elicitor remarkably stimulated the accumulation of carotenoids with a 13.72% increase compared with the control. Subsequently, the entire Rh. glutinis elicitor (part NHK) was divided into three parts, namely, exopolysaccharide (EPS) (part E), mixture of EPS and protein (part PE), and other components (part O), to analyze their effects on carotenoid accumulations. Results showed that part O may be the effective component that remarkably stimulates the biosynthesis of carotenoids with a 26% increase compared with the control. This research demonstrated that Rh. glutinis elicitor can effectively increase the content of natural carotenoids in C. militaris, and provided an important reference for the development and utilization of carotenoid industrialization.

This work investigated the antiradical, antibacterial, and antifungal properties of a methanol extract from Fuscoporia torulosa (PeruMyc1834) fruit body from Central Italy. The ITS partial sequence of the F. torulosa strain matched at a 100% rate those of various strains belonging to the species. Two complementary test systems, namely α,α-diphenyl-β-picrylhydrazyl (DPPH) radical scavenging and β-carotene/linoleic acid systems, were used. Free radical scavenging, measured by DPPH assay on methanol extract, showed an activity of 38.17 ± 2.74% of Trolox activity. The methanol extract of F. torulosa inhibited the growth of Gram-positive bacteria, particularly those belonging to the genus Bacillus (i.e., B. subtilis and B. cereus), better than Gram-negative bacteria. All the fungal isolates used in the present study for antifungal activity inhibited their growth when a concentration of 2.27 mg mL-1 of methanol extract was used (minimum inhibitory concentration from < 0.28 to 2.27 mg mL-1). This study established that the extracted antioxidant and antimicrobial compounds produced by F. torulosa can be exploited in treating diseases that affect humans, plants, and animals. There is a need to carry out further study to establish the chemical composition of the methanol extract.

We introduce the principle of a new technique to isolate glycosphingolipids (GSLs) from phospholipids. Neutral and acidic GSLs in organic solvent bind to titanium dioxide under neutral pH and can be eluted with 5 mg/ml of 2,5-dihydroxybenzoic acid in methanol. This special property is applicable for eliminating phospholipids, including sphingomyelin, which cannot be eliminated by a typical mild alkaline treatment. By using this technique, we demonstrated the rapid separation of minor components of GSLs, namely sulfatide and gangliosides from rabbit serum and liver, respectively. The minor GSL components were effectively purified despite both sources containing tremendous amount of phospholipids and simple lipids such as cholesterol, cholesteryl esters and triglycerides.

The study was designed to explore the beneficial effect of Musca domestica larvae extract (MDLE) on a metabolic disorder using a diabetic rat model. Streptozotocin-induced diabetic rats were treated with or without MDLE. Blood glucose, insulin levels, lipid profiles, and oxidative stress markers were measured. The morphological changes in the pancreas and liver were determined, as well as insulin expression. The expression of glucose transporter 4 (GLUT4), phospho-adenosine monophosphate-activated protein kinase (p-AMPK)/total AMPK, superoxide dismutase 1 (SOD1), catalase (CAT), and peroxisome proliferator-activated receptor gamma (PPARγ) were detected. Compared with untreated diabetic rats, MDLEtreated rats had decreased urine volume, food intake, and water intake, along with significantly lower levels of blood glucose, malondialdehyde (MDA), plasma triglycerides, low-density lipoprotein (LDL), and total cholesterol. MDLEtreated rats also had higher levels of SOD activity, high-density lipoprotein (HDL), and insulin. MDLE treatment partially restored the β-cell population, improved the liver necrosis and islet cell damage, reversed the decreased expression of GLUT4, phospho-AMPK, SOD1, and CAT in the liver, skeletal muscle and pancreatic tissue, and also increased the expression of PPARγ in the liver and adipose tissue in diabetic rats. In conclusion, the obtained results suggest that MDLE could possibly be used pharmacologically as an adjuvant for the treatment of diabetes.

We studied the effects of elastase, cigarette smoke extract, D-galactosamine hydrochloride, and tyrosine kinase inhibitor SU5416 on endothelial progenitor cells and angiogenesis precursors, as well as on Notch-1 expression by immature endothelial cells. Simultaneously with pulmonary emphysema, different damaging factors with diverse mechanisms of action caused pathological changes in the microvascular network of the lungs and destroyed the alveolar endothelium in female C57Bl/6 mice. D-galactosamine hydrochloride disturbed mobilization of endothelial progenitor cells expressing VEGFR (CD45-CD309 + ) and angiogenesis progenitors (CD45 - CD309 + CD117 + ) and their migration into emphysema expanded lungs. Elastase inhibited VEGFR-expressing endothelial progenitor cells, while cigarette smoke extract inhibited cells with CD45 - CD31 + CD34 + phenotype. In pulmonary emphysema provoked by elastase or D-galactosamine hydrochloride, angiogenesis was provided by endothelial cells with CD45 - CD31 + CD34 + phenotype, whereas in emphysema modeled with SU5416 or cigarette smoke extract, it was provided by the endothelial VEGFR-expressing cells and mature CD31 + endothelial cells, respectively. Replenishment of immature endothelial cells damaged by elastase and SU5416 involved Notch-1 + angiogenesis precursors and Notch-1 + endothelial progenitor cells with VEGFR.

The emergence of antimicrobial resistance to commonly used antibiotics has necessitated the development of new antimicrobial products. Crude extracts produced by actinomycetes contain antimicrobial metabolites that can inhibit bacterial growth. The objective of our study was to evaluate the antimicrobial activity of crude extracts (Caat1-54 and CaatP5-8) produced by actinomycetes against isolates of Staphylococcus aureus, Staphylococcus chromogenes, Streptococcus dysgalactiae, and Streptococcus uberis, which were obtained from the milk of cows affected by mastitis in 23 dairy herds. Twenty isolates of each bacterial species were used to define minimum inhibitory concentrations (MIC) of both crude extracts and ceftiofur (positive control). The MIC 50 and MIC 90 were defined at the concentration required to inhibit the growth of 50 and 90% of bacterial isolates tested, respectively. The MIC results were evaluated by survival analysis. Staphylococcus aureus isolates presented MIC 90 of Caat 1-54 ≥6.25 µg/mL, ceftiofur ≥12.5 µg/mL, and Caat P5-8 ≥100 µg/mL. Streptococcus uberis presented MIC 90 of ceftiofur ≥0.39 µg/mL, Caat 1-54 ≥50 µg/mL, and Caat P5-8 ≥100 µg/mL. Staphylococcus chromogenes isolated from subclinical mastitis presented MIC 90 of Caat 1-54 ≥0.78 µg/mL and ceftiofur and Caat P5-8 of ≥6.25 and ≥100 µg/mL, respectively. Streptococcus dysgalactiae isolated from clinical mastitis presented similar MIC 90 values between antimicrobials tested (ceftiofur, Caat 1-54, and Caat P-58), but these values (≥100 µg/mL) were higher than the values obtained from other pathogens evaluated in the present study. Our results indicate that Caat 1-54 and Caat P5-8 crude extracts present in vitro antimicrobial activity against isolates of Staph. aureus, Staph. chromogenes, Strep. dysgalactiae, and Strep. uberis isolated from clinical and subclinical mastitis., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In the present study, secondary metabolites from an endophytic fungus, Alternaria alternata, colonizing Carica papaya, demonstrated antiquorum sensing properties against Pseudomonas aeruginosa. This study reports the antagonistic effects of fungal crude extract of A. alternata against the various quorum sensing (QS) associated virulent factors such as percentage decrease in production of pyocyanin, alginate, chitinase and rhamnolipid; significant decrease in proteases activity such as LasA protease activity, staphylolytic activity, Las B elastase; and a marked decrease in biofilm formation and associated factors such as exopolysaccharide (EPS) production and cell surface hydrophobicity (CSH). Further, motility pattern i.e., swimming and swarming was also found to be inhibited. This down regulation of QS and associated factors are further supported by in-silico analysis of interaction between QS receptor LasR and bioactive molecules viz., sulfurous acid, 2-propyl tridecyl ester and 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester present in fungal crude extract, found based on GCMS analysis, sketches the modulating ability of QS expression. This is the first report on an endophytic fungus of C. papaya having a role in QS inhibition against P. aeruginosa and lays a platform to explore further the endophytes for potent therapeutic agents in QS., (Copyright © 2018 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

This study evaluated the use of commercial rhamnolipid biosurfactant supplementation in the phytoremediation of a soil via sunflower (Helianthus annuus L.) cultivation. The soil, obtained from an industrial area, was co-contaminated with heavy metals and petroleum hydrocarbons. The remediation tests were monitored for 90 days. The best results for removal of contaminants were obtained from the tests in which the sunflower plants were cultivated in soil with 4 mg kg -1 of the rhamnolipid. Under these conditions, reductions of 58% and 48% were obtained in the total petroleum hydrocarbon (TPH) and polycyclic aromatic hydrocarbon (PAH) concentrations, respectively; reductions in the concentrations of the following metals were also achieved: Ni (41%), Cr (30%), Pb (29%), and Zn (20%). The PCR-DGGE analysis of soil samples collected before and after the treatments verified that the plant cultivation and biosurfactants supplementation had little effect on the structure of the dominant bacterial community in the soil. The results indicated that sunflower cultivation with the addition of a biosurfactant is a viable and efficient technology to treat soils co-contaminated with heavy metals and petroleum hydrocarbons.

Conventional and commercial methods for isolation of nucleic acids are available for fungal samples including entomopathogenic fungi (EPF). However, there is not a unique optimal method for all organisms. The cell wall structure and the wide range of secondary metabolites of EPF can broadly interfere with the efficiency of the DNA extraction protocol. This study compares three commercial protocols: DNeasy® Plant Mini Kit (Qiagen), Wizard® Genomic DNA Purification Kit (Promega), and Axygen™ Multisource Genomic DNA Miniprep Kit (Axygen) and three conventional methods based on different buffers: SDS, CTAB/PVPP, and CTAB/β-mercaptoethanol versus three cell lysis procedures: liquid nitrogen homogenization and two bead-beating materials (i.e., tungsten-carbide and stainless-steel) for four representative species of EPF (i.e., Beauveria bassiana, Hirsutella citriformis, Isaria javanica, and Metarhizium anisopliae). Liquid nitrogen homogenization combined with DNeasy® Plant Mini Kit (i.e., QN) or SDS buffer (i.e., SN) significantly improved the yield with a good purity (~1.8) and high integrity (>20,000 bp) of genomic DNA in contrast with other methods, also, these results were better when compared with the two bead-beating materials. The purified DNA was evaluated by PCR-based techniques: amplification of translation elongation factor 1-α (TEF) and two highly sensitive molecular markers (i.e., ISSR and AFLP) with reliable and reproducible results. Despite a variation in yield, purity, and integrity of extracted DNA across the four species of EPF with the different DNA extraction methods, the SN and QN protocols maintained a high-quality of DNA which is required for downstream molecular applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method - TruTip - with our newly-developed mechanical lysis method. This is the first report of this method for homogenizing viscous and heterogeneous samples and lysing difficult-to-disrupt cells using "MagVor": a rotating magnet that rotates a miniature stir disk amidst glass beads confined inside of a disposable tube. Using this system, we demonstrate automated nucleic acid extraction from methicillin-resistant Staphylococcus aureus (MRSA) in nasopharyngeal aspirate (NPA), influenza A in nasopharyngeal swabs (NPS), human genomic DNA from whole blood, and Mycobacterium tuberculosis in NPA. The automated workstation yields nucleic acid with comparable extraction efficiency to manual protocols, which include commercially-available Qiagen spin column kits, across each of these sample types. This work expands the scope of applications beyond previous reports of TruTip to include difficult-to-disrupt cell types and automates the process, including a method for removal of organics, inside a compact bench-top workstation., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Characterization of complex natural product mixtures to the absolute structural level of their components often requires significant amounts of starting materials and lengthy purification process, followed by arduous structure elucidation efforts. The crystalline sponge (CS) method has demonstrated utility in the absolute structure elucidation of isolated organic compounds at miniscule quantities compared to conventional methods. In this work, we developed a new CS-based workflow that greatly expedites the in-depth structural analysis of crude natural product extracts. Using a crude extract of the red alga Laurencia pacifica, we showed that CS affinity screening prior to compound isolation enables prioritization of analytes present in the extract, and we subsequently resolved the molecular structures of six sesquiterpenes with stereochemical clarity from around 10 mg crude extract. This study demonstrates a new chemotyping workflow that can greatly accelerate natural product discovery from complex samples., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Biofilm forming pathogenic bacteria showing resistance towards antimicrobial agents has increased the urge to find an alternative treatment strategy. Among the biofilm forming pathogenic bacteria, Pseudomonas aeruginosa is a well-recognised Gram negative biofilm former causing nosocomial infection and other disease among immunocompromised patients. The aim of the current study is to evaluate the antipathogenic potentials of S. platensis against P. aeruginosa. S. platensis methanolic extract (SME) inhibited the biofilm (89%), extracellular polymeric substances (EPS), cell surface hydrophobicity (CSH) (44%), pyocyanin, pyoveridin and swarming motility of P. aeruginosa. Partial purification of SME using thin layer chromatography (TLC), column chromatography and gas chromatography mass spectroscopy (GCMS) revealed the major component as hexadecanoic acid (HxD). Further analysis through in silico approach showed the efficient binding of HxD with the biofilm regulatory proteins (Las R and salidase enzyme) of P. aeruginosa. The coating of HxD over different material surfaces efficiently prevented the adhesion of P. aeruginosa biofilm. The results of the toxicity assay revealed that the SME is non-toxic towards Artemia salina at the given concentration. Hence the overall work proves the potential non-toxic effects of SME against P. aeruginosa pathogenicity., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Maggot extract (ME) accelerates rat skin wound healing, however its effect on cell maintenance in wound tissues remains unclear. B‑cell lymphoma (Bcl) 2‑associated athanogene (BAG)3 inhibits apoptosis and promotes autophagy by associating with Bcl‑2 or Beclin 1. Bcl‑2, the downstream effector of signal transducer and activator of transcription 3 signaling, is enhanced in ME‑treated wound tissues, which may reinforce the Bcl‑2 anti‑apoptotic activity and/or cooperate with Beclin 1 to regulate autophagy during wound healing. The present study investigated expression levels of BAG3, Bcl‑2, Beclin 1 and light chain (LC)3 levels in rat skin wound tissues in the presence and absence of ME treatment. The results revealed frequent TUNEL‑negative cell death in the wound tissues in the early three days following injury, irrespective to ME treatment. TUNEL‑positive cells appeared in the wound tissues following 4 days of injury and 150 µg/ml ME efficiently reduced apoptotic rate and enhanced BAG3 and Bcl‑2 expression. Elevated Beclin 1 and LC3 levels and an increased LC3 II ratio were revealed in the ME‑treated tissues during the wound healing. The results of the present study demonstrate the anti‑apoptotic effects of BAG3 and Bcl‑2 in ME‑promoted wound healing. Beclin 1/LC3 mediated autophagy may be favorable in maintaining cell survival in the damaged tissues and ME‑upregulated BAG3 may enhance its activity.

Bacterial diseases are widespread in aquaculture farms and causative agents often adapt to biofilm mode of growth. These biofilms are detrimental to aquaculture species as they resist antibiotics and other agents that are used to control them. Two bacterial pathogens isolated from infected prawn samples were identified as Vibrio alginolyticus and Pseudomonas gessardii on the basis of morphological features, biochemical characteristics, 16S r RNA gene sequencing and phylogenetic analysis. Their pathogenic nature was confirmed by performing in vivo challenge experiments using Artemia salina as a model system. Seven days post infection, the mortality observed with V. alginolyticus and P. gessardii was 97 ± 4.08% and 77.5 ± 5.24%, respectively. The isolates formed extensive biofilms on polystyrene and glass surfaces. These infections could be controlled in an effective manner by using the cell free supernatant (CFS) of a tropical marine epizoic strain of Bacillus licheniformis D1 that is earlier reported to contain an antimicrobial protein (BLDZ1). The CFS inhibited biofilms in an efficient manner (82.35 ± 1.69 and 82.52 ± 1.11% for V. alginolyticus and P. gessardii, respectively) on co-incubation. In addition, pre-formed biofilms of V. alginolyticus and P. gessardii were also removed (84.53 ± 1.26 and 67.08 ± 1.43%, respectively). Fluorescence and scanning electron microscopic studies confirmed the antibiofilm potential of this protein on glass surfaces. The antibiofilm nature was due to the anti-adhesion and antimicrobial properties exhibited by the CFS. Treatment of A. salina with CFS (6 h prior to infections) was effective in protecting larvae against infections by field isolates. This study highlights the significance of marine natural products in providing alternative biofilm controlling agents to tackle infections and decreasing the usage of antibiotics in aquaculture settings., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Marine microalgae are considered a potentially new and valuable source of biologically active compounds for applications in several biotechnology sectors. They can be easily cultured, have short generation times and enable an environmentally-friendly approach to drug discovery by overcoming problems associated with the over-utilization of marine resources and the use of destructive collection practices. Considering the increasing rate of antibiotic-resistance bacteria and infections by fungi, 46 microalgae have been screened in this study for possible antibacterial and antifungal activities. Two different extraction methods have been used in order to increase the probability of finding positive hits. In particular, we screened microalgae in both control and nutrient stress conditions. We also tested different strains for 7 species in order to study potentially different bioactivities due to strain diversity. Results showed that extracts of two diatoms, Skeletonema tropicum and Chaetoceros pseudocurvisetus, had anti-tuberculosis activity and were active only when cultured in the control and phosphate-starvation conditions, while the nitrogen starvation condition showed no activity. In addition, we tested both the organic and water extracts and found that only the organic extracts for both diatoms were active. The organic extracts of these two diatom species were not toxic on normal human cell lines.

Mosquito control using chemical insecticides is facing a major challenge due to development of insecticide resistance. Improving the efficiency of existing insecticides using synergistic secondary metabolites of biological origin is increasingly being researched. Herein, we evaluated the toxicity of Fusarium oxysporum extract alone and in binary combinations with temephos, on larvae and pupae of Anopheles stephensi, Aedes aegypti and Culex quinquefaciatus. F. oxysporum extract was characterized using TLC, FT-IR and GC-MS. After 24 h of exposure, the binary combination of temephos + F. oxysporum extract (1:1 ratio) was highly toxic to larvae of An. stephensi (LC 50 : 35.927 μg/ml), Ae. aegypti (LC 50 : 20.763 μg/ml) and Cx. quinquefasciatus, (LC 50 : 51.199 μg/ml). For pupae LC 50 values were 38.668, 26.394, and 72.086 μg/ml, respectively. Histology studies of mosquitoes exposed to F. oxysporum extract showed vacuolation in epithelium, as well as in adipose, and muscle tissues of larval midgut. Overall, our results show that the synergistic combination of temephos and F. oxysporum extract is highly effective to control mosquito young instars.

Increased resistance to the first-line treatment against P. falciparum malaria, artemisinin-based combination therapies, has been reported. Here, we tested the effect of crude ethanolic extract of the fungus Trichoderma stromaticum (Ext-Ts) on the growth of P. falciparum NF54 in infected human red blood cells (ihRBCs) and its anti-malarial and anti-inflammatory properties in a mouse model of experimental cerebral malaria. For this purpose, ihRBCs were treated with Ext-Ts and analysed for parasitaemia; C57BL/6 mice were infected with P. berghei ANKA (PbA), treated daily with Ext-Ts, and clinical, biochemical, histological and immunological features of the disease were monitored. It was observed that Ext-Ts presented a dose-dependent ability to control P. falciparum in ihRBCs. In addition, it was demonstrated that Ext-Ts treatment of PbA-infected mice was able to increase survival, prevent neurological signs and decrease parasitaemia at the beginning of infection. These effects were associated with systemically decreased levels of lipids and IFN-γ, ICAM-1, VCAM-1 and CCR5 cerebral expression, preserving blood brain barrier integrity and attenuating the inflammatory lesions in the brain, liver and lungs. These results suggest that Ext-Ts could be a source of immunomodulatory and antimalarial compounds that could improve the treatment of cerebral malaria.

Gelidium elegans extract (GEE) is derived from a red alga from the Asia-Pacific region, which has antioxidant, anti-adipogenic, and anti-hyperglycemic effects. However, detailed studies of the toxicology of GEE have not been performed. We evaluated the single oral dose toxicity of GEE in male and female Sprague-Dawley (CD) rats. GEE did not cause deaths or have toxic effects at dosages of 5000 mg/kg/day, although compound-colored stools and diarrhea were observed in both sexes, which lasted <2 days. Therefore, the LD 50 of GEE is likely to be >5000 mg/kg. We next evaluated the repeated oral dose toxicity of GEE in CD rats over 14 days and 13 weeks. GEE did not induce any significant toxicological changes in either sex at 2000 mg/kg/day. Repeated oral dose toxicity studies showed no adverse effects, in terms of clinical signs, mortality, body mass, food consumption, ophthalmic examination, urinalysis, hematology, serum biochemistry, necropsy, organ masses, or histopathology, at dosages of 500, 1000, or 2000 mg/kg/day. The no observed adverse effect level (NOAEL) for GEE is thus likely to be >2000 mg/kg/day, and no pathology was identified in potential target organs. Therefore, this study indicates that repeated oral dosing with GEE is safe in CD rats., Competing Interests: The authors declare no conflict of interest.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are the most valuable source of autologous cells for transplantation and tissue regeneration to treat osteoporosis. Although BM-MSCs are the primary cells responsible for maintaining bone metabolism and homeostasis, their regenerative ability may be attenuated in postmenopausal osteoporosis patients. Therefore, we first examined potential abnormalities of BM-MSCs in an oestrogen-deficient rat model constructed by ovariectomy (OVX-MSCs). Cell proliferation, mobilisation, and regulation of osteoclasts were downregulated in OVX-MSCs. Moreover, therapeutic effects of OVX-MSCs were decreased in OVX rats. Accordingly, we developed a new activator for BM-MSCs using human umbilical cord extracts, Wharton's jelly extract supernatant (WJS), which improved cell proliferation, mobilisation and suppressive effects on activated osteoclasts in OVX-MSCs. Bone volume, RANK and TRACP expression of osteoclasts, as well as proinflammatory cytokine expression in bone tissues, were ameliorated by OVX-MSCs activated with WJS (OVX-MSCs-WJ) in OVX rats. Fusion and bone resorption activity of osteoclasts were suppressed in macrophage-induced and primary mouse bone marrow cell-induced osteoclasts via suppression of osteoclast-specific genes, such as Nfatc1, Clcn7, Atp6i and Dc-stamp, by co-culture with OVX-MSCs-WJ in vitro. In this study, we developed a new activator, WJS, which improved the functional abnormalities and therapeutic effects of BM-MSCs on postmenopausal osteoporosis.

This study evaluates the in vitro inhibition of angiotensin-converting enzyme (ACE) and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA) by Pleurotus pulmonarius extracts. The protective effect on the endothelial membrane against oxidative stress through the protection of nitric oxide bioavailability, as well as inhibition of endocan expression, was evaluated using human aortic endothelial cells (HAECs). Crude cold aqueous extract exhibited the most potent inhibitory activities against ACE and HMG-CoA reductase, with 61.79% and 44.30% inhibition, respectively. It also protected the bioavailability of NO released by HAECs, with 84.88% cell viability. The crude hot water extract was the most potent in inhibiting endocan expression, with 18.61% inhibition.

Kombucha is a nonalcoholic beverage traditionally made by fermenting black tea using a combination of yeast and acetic acid bacteria (AAB) cultures. Ganoderma lucidum hot water extract (HWE) was used-to our knowledge for the first time-to prepare a novel, health-promoting kombucha product. During the 11-day fermentation, pH, total acidity, and the numbers of yeasts and AAB were monitored. It was found that sweetened G. lucidum HWE was a good medium for yeast and AAB growth. The desired acidity for the beverage was reached on the second day (3 g/L) of the fermentation process; the maximum established acidity was 22.8 ± 0.42 g/L. Fourier transform infrared analysis revealed that the vacuum-dried beverage is a mixture of various compounds such as polysaccharides, phenols, proteins, and lipids. Total phenolic content of the liquid sample was 4.91 ± 0.2338 mg gallic acid equivalents/g, whereas the vacuum-dried sample had a smaller amount of phenolics (2.107 ± 0.228 mg gallic acid equivalents/g). Established half-maximal effective concentrations for DPPH scavenging activity and reducing power were 22.8 ± 0.17 and 10.61 ± 0.34 mg/mL, respectively. The antibacterial testing revealed that activity does not originate solely from synthesized acetic acid. The liquid G. lucidum beverage was the most effective against the tested bacteria, with the lowest minimum inhibitory concentration (0.04 mg/mL) against Staphylococcus epidermidis and Rhodococcus equi, and a minimum bactericidal concentration (0.16 mg/mL) against Bacillus spizizenii, B. cereus, and R. equi. The vacuum-dried sample was less effective, with the lowest minimum bactericidal concentration against the Gram-positive bacteria R. equi (1.875 mg/mL) and against the Gram-negative bacteria Proteus hauseri (30 mg/mL).

Despite considerable recent work to reveal different features of mushrooms species, the few studies of antiviral activities are inadequate and therefore further studies are required. Morchella conica, M. esculenta, Terfezia boudieri, Pleurotus ostreatus, Tricholoma anatolicum, Fomes fomentarius, Laetiporus sulphureus, Phellinus igniarius, Porodaedalea pini, and Pyrofomes demidoffii from Turkey were investigated to reveal their in vitro cytotoxic and anti-herpes simplex virus 1 (HSV-1) activities. The crude methanol extracts (MEs) and aqueous extracts (AEs) of fungal species and acyclovir (ACV) were used. Various dilutions were used to assess the cytotoxic effects of fungal species (50-0.10 mg/mL) and ACV (500-0.98 μg/mL) on uninfected Vero cells. Maximum nontoxic concentrations were determined for all extracts and ACV by comparing the optical densities of their cell controls. The concentration providing 50% protection against the cytopathic effect caused by the virus, extracts, and ACV (EC50) was calculated, and the half-maximal cytotoxic concentration (CC50) and the selectivity index (SI) were determined, the latter as the ratio of CC50 to EC50. While the AEs of F. fomentarius (EC50, 11.22 mg/mL; SI > 4.46), Ph. igniarius (EC50, 9.71 mg/mL; SI > 5.15), and P. pini (EC50, 7.16 mg/mL; SI > 6.98) showed considerable antiherpetic activity, MEs and AEs of the other fungal species did not showed any effects. The EC50 and SI of ACV were determined as 0.20 μg/mL and 3085, respectively. The results demonstrate that F. fomentarius, Ph. igniarius, and P. pini have important anti-HSV-1 activity.

Auricularia delicata, an edible mushroom, has been used as a traditional medicine in Manipur, India, for various gastrointestinal and liver ailments. This study evaluates the antioxidant, antimicrobial, and hepatoprotective potential of A. delicata. A. delicata fruiting bodies were extracted with hexane, chloroform, ethyl acetate, and methanol. All these extracts were examined for in vitro antioxidant activity. To study antimicrobial activity, minimum inhibitory concentrations (milligrams per milliliter) were determined through the use of the broth dilution method. In vivo hepatoprotective activity against acetaminophen-induced hepatic injury in rats was investigated by evaluating serum biochemistry, antioxidant enzymes, and histopathology. With regard to antioxidant activity, 21 and 48 μg/mL were the lowest half-maximal effective concentrations, obtained for the methanol and ethyl acetate fractions, respectively. In the antimicrobial study, the ethyl acetate fraction showed the lowest inhibition of Bacillus subtilis, Enterococcus feacium, Streptococcus aureus, B. cereus, and Escherichia coli, with minimum inhibitory concentrations of 0.03, 0.015, 0.03, 0.11, and 0.5 mg/mL, respectively. Further, in in vivo studies, elevated levels of biochemical markers were significantly returned to near normal values; this was supported by histopathological changes. Thus A. delicata showed antimicrobial, antioxidant, and protective roles in induced hepatic injury. Phytochemical analysis using high-performance liquid chromatography revealed the presence of chlorogenic acid in the extracts. Its protective property might be due to the presence of a mitochondrial-targeted antioxidant effect of chlorogenic acid. The antimicrobial activity accounts for its use against diarrhea. Hence, A. delicata could be one of the best sources for natural gastrointestinal and hepatoprotective medicines in the future.

This protocol details protein extraction from mouse tissues for immunoprecipitation purposes and has been applied for the performance of large-scale immunoprecipitations of target proteins from various tissues for the identification of associated proteins by mass spectroscopy. The key factors in performing a successful immunoprecipitation directly relate to the abundance of target protein in a particular tissue type and whether or not the embryonic, newborn, or adult mouse-derived tissues contain fibrous and other insoluble material. Several tissue types, including lung and liver as well as carcinomas, contain significant amounts of fibrous tissue that can interfere with an immunoprecipitation., (© 2017 Cold Spring Harbor Laboratory Press.)

This protocol describes the lysis of tissue culture cells for the solubilization of proteins of interest for immunoprecipitation. Upon collection of cells by centrifugation and depending on the use of either Tris- or phosphate-based cell lysis buffers, cells are rinsed, respectively, in either TBS or PBS before lysis. If possible, the pH of the Wash buffer should match that of the Lysis buffer. Adherent cells can be directly lysed on the plate. This is particularly useful upon lysis encompassing mild nonionic detergents, leaving the cytoskeleton intact. Alternatively, adherent cells can be scraped off the plate and directly resuspended in Lysis buffer or Wash buffer for transfer to a tube followed by the addition of Lysis buffer., (© 2017 Cold Spring Harbor Laboratory Press.)

We demonstrate on-chip, differential DNA and RNA extraction from a single cell using a microfluidic chip and a two-stage lysis protocol. This method enables direct use of the whole extract, without additional washing steps, reducing sample loss. Using this method, the tumor driving pathway in individual cells from a colorectal cancer cell line was determined by applying a Bayesian computational pathway model to sequences obtained from the RNA fraction of a single cell and, the mutations driving the pathway were determined by analyzing sequences obtained from the DNA fraction of the same single cell. This combined functional and mutational pathway assessment of a single cell could be of significant value for dissecting cellular heterogeneity in tumors and analyzing single circulating tumor cells.

Objective: Over 10-15% of couples in different countries are infertile. Male infertility is a contributing factor and the only cause of infertility in respectively 50% and 20-30% of all cases of infertility. According to previous research, micro-elements isolated from date palm pollen (DPP), e.g. estrogen and sterols, may enhance male and female fertility. DPP has also been reported to improve sperm parameters including sperm motility and viability, acrosome reaction, and lipid peroxidation. This article may justify the need for a future systematic review and meta-analysis about the effects of DPP on the reproductive system and DPP's ability to enhance fertility. It will then describe the methodology of such a study., Main Text: A comprehensive search of relevant randomized and quasi-randomized controlled trials will be performed in MEDLINE, EMBASE, Web of Science, Cochrane Central, ProQuest, and Google Scholar databases. Two authors will independently assess the eligibility of the studies and consult the third author in cases of disagreement. The risk of bias of the randomized controlled trials and animal studies will be evaluated using the Cochrane risk of bias tool and the Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) risk of bias tool, respectively. This study will raise no ethical issues as it will review the findings of previous research. The results are intended to be published in a peer-reviewed medical journal.

Multidrug-resistance bacteria commonly use cell-to-cell communication that leads to biofilm formation as one of the mechanisms for developing resistance. Quorum sensing inhibition (QSI) is an effective approach for the prevention of biofilm formation. A Gram-negative bacterium, Delftia tsuruhatensis SJ01, was isolated from the rhizosphere of a species of sedge ( Cyperus laevigatus ) grown along the coastal-saline area. The isolate SJ01 culture and bacterial crude extract showed QSI activity in the biosensor plate containing the reference strain Chromobacterium violaceum CV026. A decrease in the violacein production of approximately 98% was detected with the reference strain C. violaceum CV026. The bacterial extract (strain SJ01) exhibited anti-quorum sensing activity and inhibited the biofilm formation of clinical isolates wild-type Pseudomonas aeruginosa PAO1 and P. aeruginosa PAH. A non-toxic effect of the bacterial extract (SJ01) was detected on the cell growth of the reference strains as P. aeruginosa viable cells were present within the biofilm. It is hypothesized that the extract (SJ01) may change the topography of the biofilm and thus prevent bacterial adherence on the biofilm surface. The extract also inhibits the motility, virulence factors (pyocyanin and rhamnolipid) and activity (elastase and protease) in P. aeruginosa treated with SJ01 extract. The potential active compound present was identified as 1,2-benzenedicarboxylic acid, diisooctyl ester. Microarray and transcript expression analysis unveiled differential expression of quorum sensing regulatory genes. The key regulatory genes, LasI, LasR, RhlI , and RhlR were down-regulated in the P. aeruginosa analyzed by quantitative RT-PCR. A hypothetical model was generated of the transcriptional regulatory mechanism inferred in P. aeruginosa for quorum sensing, which will provide useful insight to develop preventive strategies against the biofilm formation. The potential active compound identified, 1,2-benzenedicarboxylic acid, diisooctyl ester, has the potential to be used as an anti-pathogenic drug for the treatment of biofilm-forming pathogenic bacteria. For that, a detailed study is needed to investigate the possible applications.

Antrodia cinnamomea (AC) exhibits many bioactivities, including anti-inflammatory, anti-cancer, and hepatoprotection activities. Many researchers have studied the functions of the components or fractions of AC, but the functions of the original extractions of AC have not been studied. In addition, the detailed relationship between AC and immune-related signaling pathways is unclear. In this study, we screened the effects of CCM111, which is the extract of AC, on seven immune-related signaling pathways and further investigated whether CCM111 can influence inflammation. Interestingly, our results showed that CCM111 significantly inhibited the IL-6-stimulated STAT3 pathway and the LPS-stimulated NF-κB pathway in macrophages. CCM111 also decreased the phosphorylation of STAT3, Tyk2 and the nuclear translocation of p65. Moreover, CCM111 and F4, a fraction of CCM111, down-regulated nitric oxide (NO) production, the protein levels of iNOS and COX-2, and inflammatory cytokines in macrophage cells. Therefore, our study suggested that CCM111 has the potential to be developed as an effective anti-inflammatory agent.

Lichen-forming fungi are known to have various biological activities, such as antioxidant, antimicrobial, antitumor, antiviral, anti-inflammation, and anti proliferative effects. However, the immunosuppressive effects of Bryoria sp. extract (BSE) have not previously been investigated. In this study, the inhibitory activity of BSE on the proliferation of CD8 + T cells and the mixed lymphocytes reaction (MLR) was evaluated in vitro. BSE was non-toxic in spleen cells and suppressed the growth of splenocytes induced by anti-CD3. The suppressed cell population in spleen cells consisted of CD8 + T cells and their proliferation was inhibited by the treatment with BSE. This extract significantly suppressed the IL-2 associated with T cell growth and IFN-γ as the CD8 + T cell marker. Furthermore, BSE reduced the expression of the IL-2 receptor alpha chain (IL-2Rα) on CD8 + T cells and CD86 on dendritic cells by acting as antigen-presenting cells. Finally, the MLR produced by the co-culture of C57BL/6 and MMC-treated BALB/c was suppressed by BSE. IL-2, IFN-γ, and CD69 on CD8 + T cells in MLR condition were inhibited by BSE. These results indicate that BSE inhibits the MLR via the suppression of IL-2Rα expression in CD8 + T cells. BSE has the potential to be developed as an anti-immunosuppression agent for organ transplants.

Seven new cucurbitane glucosides, 11-oxomogrosides III E and IV (1 and 2), 11-oxoisomogroside V (3), 7-oxomogrosides III E and IV (4 and 5), and mogrosides VI A and VI B (6 and 7), were separated from the crude extract of Siraitia grosvenorii. The new structures were defined by analysis of their 1 H and 13 C NMR, 2D NMR, and HRESIMS data. Especially, the band-selective constant time HSQC and band-selective constant time HMBC techniques were recuited to elucidate the structures of the complex glucoside moieties. Using the PGC-1α promoter driven luciferase reporter assay, the isolated compounds were examined for PGC-1α promoter activity.

We investigated the beneficial effects of the crude Ecklonia cava flake (CEF), which is a residual product after polyphenol extraction from Ecklonia cava , on inflammation in LPS-stimulated RAW264.7 cells. A group of five different CEF extracts was obtained by a preparation process using water, hydrochloric acid or temperature. We observed that large-size (>19 kDa) CEF extract, which was extracted with water at 95 °C (CEF-W, 95 °C), suppressed the production of inflammatory cytokines by inhibiting its mRNA expression in LPS-induced RAW264.7 cells. TLR4 signaling involvements were negatively regulated by CEF-W, 95 °C. CEF-W, 95 °C repressed the translocation of NF-κB from cytoplasm into nucleus in LPS-induced RAW264.7 cells. CEF-W, 95 °C attenuated the phosphorylation of TBK1 and IRF3 by inhibiting the phosphorylation of ERK. Taken together, we demonstrated that large-size CEF-W, 95 °C may act as a negative regulator of inflammation through the suppression of TLR4 signaling constituents in LPS-induced RAW264.7 cells.

Strain KMM 9513 T was isolated from a sediment sample collected from the Sea of Japan seashore and selected due to its ability to inhibit indicator bacterial growth. The strain KMM 9513 T has been recently described as a novel species Rheinheimera japonica. This study was undertaken to determine which substances produced by strain KMM 9513 T could be responsible for its antimicrobial activity. Eight compounds were obtained from an ethyl acetate extract of R. japonica KMM 9513 T . The structures of five diketopiperazines (4-8) and diisobutyl-, dibutyl- and bis(2-ethylhexyl) phthalates (1-3) were established on the basis of detailed interpretation of NMR data, by Marfey method and optical rotation data. The structures of diketopiperazines were determined as cyclo-(L-valyl-L-proline), cyclo-(L-valyl-D-proline), cyclo-(L-phenylalanyl-L-proline), cyclo-(L-leucyl-L-proline), and cyclo-(L-phenylalanyl-D-proline). Compounds 1-3, 5 and 8 revealed antimicrobial activities against Bacillus subtilis and/or Enterococcus faecium and Staphylococcus aureus. In this paper, we describe the isolation and structural elucidation of the isolated compounds 1-8. This is the first report of the characterisation of low molecular weight antibacterial metabolites produced by a member of the genus Rheinheimera.

G-protein coupled receptors are the largest family of integral membrane proteins found within the human genome. They function as receptors and modulators to a wide range of ligands and responses which are crucial for human health. GPCR study, specifically the investigation of structure and interaction to cognate ligands, is of high priority. Limitations for structural study can be traced in part, to obtaining suitable quantities of recombinant protein. We sought to address the limitations of traditional recombinant technologies by utilising an Escherichia coli based cell-free protein synthesis (CFPS) approach for production of a thermostable neurotensin receptor 1 (en2NTS 1 ). Initial results were promising, with a high amount (up to 2 mg/mL) of en2NTS 1 produced, that had attained correct secondary structure. Meanwhile, concurrent experiments indicated that CFPS produced en2NTS 1 showed non-competitive binding to the peptide ligand neurotensin8-13 when compared to E. coli produced en2NTS 1 . 1 H- 13 C HMQC SOFAST NMR spectra were indicative of disrupted tertiary structure for CFPS produced 13 CH 3 -methionine labelled en2NTS 1 . The results obtained, indicate CFPS produced en2NTS 1 is not forming a discrete tertiary structure and that further development of the CFPS technique needs to be carried out.

Shuang-Huang-Lian injectable powder (SHL)-a classical purified herbal preparation extracted from Scutellaria baicalensis , Lonicera japonica , and Forsythia suspense -has been used against human adenovirus III (HAdV₃) for many years. The combination herb and its major bioactive compounds, including chlorogenic acid, baicalin, and forsythia glycosides A, are effective inhibitors of the virus. However, no comprehensive studies are available on the antiviral effects of SHL against HAdV₃. Moreover, it remains unclear whether the mixture of chlorogenic acid, baicalin, and forsythia glycosides A (CBF) has enhanced antiviral activity compared with SHL. Therefore, a comparative study was performed to investigate the combination which is promising for further antiviral drug development. To evaluate their antivirus activity in parallel, the combination ratio and dose of CBF were controlled and consistent with SHL. First, the fingerprint and the ratio of CBF in SHL were determined by high performance liquid chromatography. Then, a plaque reduction assay, reverse transcription polymerase chain reaction (PCR), real-time polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay (ELISA) were used to explore its therapeutic effects on viral infection and replication, respectively. The results showed that SHL and CBF inhibited dose- and time-dependently HAdV₃-induced plaque formation in A549 and HEp-2 cells. SHL was more effective than CBF when supplemented prior to and after viral inoculation. SHL prevented viral attachment, internalization, and replication at high concentration and decreased viral levels within and out of cells at non-toxic concentrations in both cell types. Moreover, the expression of tumor necrosis factor alpha (TNF)-α, interleukin (IL)-1ß, and IL-6 was lower and the expression of interferon (IFN)-γ was higher in both cell types treated with SHL than with CBF. In conclusion, SHL is much more effective and slightly less toxic than CBF.

We studied activation of macrophages with humic acids extracted from peat of large deposits in the Tomsk region by two extraction methods: by hydroxide or sodium pyrophosphate. Humic acid of lowland peat types containing large amounts of aromatic carbon, phenolic and alcohol groups, carbohydrate residues and ethers, irrespectively of the extraction methods contained LPS admixture that probably determines their activating properties. Humic acid of upland peat types characterized by high content of carbonyl, carboxyl, and ester groups enhance NO production and reduce arginase expression, but these effects were minimized when sodium hydroxide was used as an extraction solvent. Pyrophosphate samples of the upland peat types were characterized by aromaticity and diversity of functional groups and have a significant advantage because of they induce specific endotoxin-independent stimulating action on antigen presenting cells.

Objective: To review the topic of fungal raw materials used for the production of allergen extracts and the associated challenges and highlight candidate areas for development before standardized fungal allergen extracts can be commercially produced., Data Sources: A PubMed search was performed using focused keywords and combined with a review of regulatory documents and industry guidelines. Several books on mycology also were consulted., Study Selections: The information obtained through the literature, books, and industry was scrutinized and combined with personal experience and expertise to write this article., Results: Fungi are complex ubiquitous organisms on Earth. They are beneficial and detrimental for humans. Fungi can cause hypersensitivity reactions, including types I, III, and IV. The procurement of fungal raw materials to prepare allergen extracts for diagnosis and possible allergen immunotherapy is complex owing to the intrinsic nature of fungi and their complex genome. Allergen manufacturers produce allergen extracts with variable qualitative and quantitative compositions, which can lead to unpredictable clinical outcomes., Conclusion: The clinician should be aware of the factors responsible for the qualitative and quantitative compositions of fungal allergen extracts and the reasons that currently preclude their standardization. Scientific advances and collaboration and cooperation between allergen manufacturing companies and regulatory agencies are necessary to improve the quality and consistency of fungal extracts. Moreover, clinicians should understand the limitations of currently available fungal extracts., (Copyright © 2016 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

Aim: To investigate antifungal potential of Xylaria sp. BIOTEC culture collection (BCC) 1067 extract against the model yeast Saccharomyces cerevisiae., Materials & Methods: Antifungal property of extract, reactive oxygen species levels and cell survival were determined, using selected deletion strains., Results: Extract showed promising antifungal effect with minimal inhibitory concentration 100 and minimal fungicidal concentration of 500 and 1000 mg/l, respectively. Strong synergy was observed with fractional inhibitory concentration index value of 0.185 for the combination of 60.0 and 0.5 mg/l of extract and ketoconazole, respectively. Extract-induced intracellular reactive oxygen species levels in some oxidant-prone strains and mediated plasma membrane rupture. Antioxidant regulator Yap1, efflux transporter Pdr5 and ascorbate were pivotal to protect S. cerevisiae from extract cytotoxicity., Conclusion: Xylaria sp. BCC 1067 extract is a potentially valuable source of novel antifungals.

Using the model of dark stress caused by animal maintenance at dimmed light we showed that Trametin (product obtained during liquid-phase culturing of Trametes pubescens xylotroph fungi) effectively prevented oxidative stress under conditions of light deprivation. The preparation increased the level of unsaturation of lipids, reduced the concentration of primary and end-products of LPO, and increases both the integral parameter of the antioxidant defense system (total antioxidant activity) and its components (activity of superoxide dismutase and reduced glutathione).

Aim: Study spectrum and expressiveness of antibacterial and antifungal properties oflichen extracts., Materials and Methods: Antimicrobial activity of acetone extracts from Hypogymnia physodes, Xanthoria parietina, Evernia prunastri, Ramalina pollinaria, Cladonia arbuscula li- chens was determinedby micro-dilution methods in broth for 4 -500 mcg/ml concentrations against 13 strains from ATCC collection and 6 clinical isolates., Results: High antibacterial activity of H. physodes and C. arbuscular extracts against staphylococci and enterococci was detected (MIC 31 - 62 mcg/ml). Antimicrobial activity against enterobacteria and Pseudo- monas aeruginosa was absent for all the extracts. E.prunastri, H.physodes and C. arbuscula extracts were active against Stenotrophomonas maltophilia strains (MIC 250 - 500 mcg/ml). Antifungal activity (MIC 500 mcg/ml for 4 Candida strains) was only detected for the E. prunastri extract., Conclusion: H.physodes and C. arbuscula lichens can be examined as a per- spective source of antibacterial substances, effective against antibiotics resistant staphyloco- cci, streptococci and S. maltophilia strains.

This work aimed to establish an effective approach to evaluate the quality of frozen fish, focusing on changes in fish tissue structure and chemical composition during storage. Fresh tilapia samples were treated by coating with tangerine peel (TP) extract and then stored at -4, -8 and -18 °C, respectively, for 40 days. The frozen fish tissues were analyzed for structural and chemical changes. Fractal dimension, which quantifies the porous structure formed in the tissue samples, texture properties including hardness and springiness, and moisture content and water activity all decreased during the storage, while the extents of lipid oxidation, measured as peroxide value and thiobarbituric acid concentration, and protein degradation, monitored with total volatile basic nitrogen and trichloroacetic acid soluble peptides, increased. The change rates of these parameters decreased with decreasing the storage temperature and by applying TP extract. A model was developed for predicting fractal dimension, which indicated the quality of preserved tilapia and thus can be used to predict the shelf life under different storage temperatures. The results demonstrated that TP extract could extend the shelf life of frozen tilapia by 35-45% by inhibiting changes in tissue structure, moisture loss, lipid oxidation and protein degradation during frozen storage., Competing Interests: The authors declare no competing financial interests.

In cases of fungal infection of the bloodstream, rapid species identification is crucial to provide adapted therapy and thereby ameliorate patient outcome. Currently, the commercial Sepsityper kit and the sodium-dodecyl sulfate (SDS) method coupled with MALDI-TOF mass spectrometry are the most commonly reported lysis protocols for direct identification of fungi from positive blood culture vials. However, the performance of these two protocols has never been compared on clinical samples. Accordingly, we performed a two-step survey on two distinct panels of clinical positive blood culture vials to identify the most efficient protocol, establish an appropriate log score (LS) cut-off, and validate the best method. We first compared the performance of the Sepsityper and the SDS protocols on 71 clinical samples. For 69 monomicrobial samples, mass spectrometry LS values were significantly higher with the SDS protocol than with the Sepsityper method (P < .0001), especially when the best score of four deposited spots was considered. Next, we established the LS cut-off for accurate identification at 1.7, based on specimen DNA sequence data. Using this LS cut-off, 66 (95.6%) and 46 (66.6%) isolates were correctly identified at the species level with the SDS and the Sepsityper protocols, respectively. In the second arm of the survey, we validated the SDS protocol on an additional panel of 94 clinical samples. Ninety-two (98.9%) of 93 monomicrobial samples were correctly identified at the species level (median LS = 2.061). Overall, our data suggest that the SDS method yields more accurate species identification of yeasts, than the Sepsityper protocol., (© The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Malaria is one of the most life-threatening health problems worldwide and treatment has been compromised by drug resistance. Identifying lead molecules from natural products might help to find better anti-malarial drugs, since those obtained from natural sources are still effective against malarial parasites. This study aimed at investigating the in vivo antiplasmodial activity of crude extract of the leaves of Ajuga remota together with its safety in mice models., Methods: In vivo parasite growth inhibitory effect of crude extract was assessed in mice inoculated with Plasmodium berghei (ANKA strain). The in vivo antiplasmodial activity of the test extract was performed against early infection (4-day suppressive test), curative effect against established infection and prophylactic effect against residual infection. Acute and sub-acute toxicity were carried out according to OECD guidelines., Results: In vivo parasite growth inhibition effect of hydroethanolic crude extract of A. remota was evaluated at 30, 50 and 100 mg/kg dose levels. It suppressed parasitaemia by 77.34% at 100 mg/kg dose level in the 4-day test. In curative and prophylactic potential tests, it suppressed parasitaemia by 66.67 and 59.66% at 100 mg/kg dose level, respectively. In vivo toxicity tests revealed no toxicity. All parasitaemia suppressions were statistically significant at P < 0.05 as compared to the vehicle-treated group. The crude extract also prolonged survival time in a dose dependent manner., Conclusions: The investigation results suggest that the leave extract of Ajuga remota possesses antimalarial activity.

Four extract fractions from the fruiting bodies of Phellinus baumii were acquired. Preliminary investigation of radical scavenging ability showed that the ethyl acetate extract exhibited the strongest antioxidant activity. To gain insight into the antioxidant mechanism, 5 antioxidant constituents were prepared from the ethyl acetate extract using high-speed countercurrent chromatography with a solvent system consisting of n-hexane, ethyl acetate, methanol, and water (0.8:3:1.5:2.5, v/v/v/v), and preparative high-performance liquid chromatography. Electrospray ionization mass spectrometry and nuclear magnetic resonance spectra were used to analyze the structures of these compounds and revealed that they were caffeic acid (compound 1), inoscavin A (compound 2), 3,4-dihydroxy benzaldehyde (compound 3), coumarin (compound 4), and naringenin (compound 5). Compound 2 showed strong antioxidant activity, and compounds 1, 3, and 5 showed moderate antioxidant activity. Thus, a more clearly and usefully scientific background is provided herein for the development of relative functional foods or biomedical products using Ph. baumii as the raw material.

Pleurotus citrinopileatus is an edible medicinal mushroom rich in biomolecules and thus has a high potential for use in formulating pharmaceutical and nutraceutical products. To test its effect on body weight and glucose control, we generated diet-induced obese (DIO) C57BL/6J male mice by feeding the mice a high-fat diet (60% fat) for 8 weeks and treating them with an ethanolic P. citrinopileatus extract (PCE) at either 200 or 500 mg/kg body weight for 12 additional weeks. The results showed that PCE significantly inhibited high-fat diet-induced weight gain, fat accumulation, and glucose intolerance in the DIO mice. Moreover, the PCE had a beneficial effect on liver and kidney function. On the basis of these results, we conclude that PCE is effective in the treatment of metabolic syndrome and thus could be a good candidate for use in future pharmaceutical or nutraceutical applications.

Despite the great advances in chemotherapeutics, infectious diseases are still one of the leading causes of death worldwide. Among some of the clinically relevant pathogens, methicillin-resistant Staphylococcus aureus (MRSA) ranks as one of the most difficult bacteria to treat. It is a common cause of skin, soft-tissue, and endovascular infections, as well as pneumonia, septic arthritis, endocarditis, osteomyelitis, and sepsis. The research on Basidiomycota is extensive; many species show a broad spectrum of pharmacological activities, including antimicrobial activity. The vast majority of the literature to date generally focuses on screening the antibacterial properties of mushroom extracts. A gap still exists in the identification of the individual compounds responsible for these properties, and few low molecular weight compounds have been described. Gymnopilus junonius, the big laughter mushroom, grows wild in Uruguay, especially on Eucalyptus spp. plantations; it is known as the "eucalyptus fungus." In this work, we report the bioguided isolation, structural elucidation, and antistaphylococcal activity of the main antimicrobial components of fresh basidiocarps of G. junonius.

The aim of this work was to study the bioactivity of crude aqueous and ethanolic extracts of Boletus edulis prepared from caps and stipes of wild-growing basidiocarps collected from the Prijepolje region (western Serbia). The bioactivity screening included antioxidant (2,2-diphenyl-l-picrylhydrazyl [DPPH], nitric oxide, super-oxide anion*, and hydroxyl radicals and ferric-reducing antioxidant power) and antiproliferative MTT assays (human breast MCF-7 cancer cell line). In addition, all extracts were primarily characterized by ultraviolet/visible spectrophotometry to determine total phenolic and flavonoid contents. The highest anti-DPPH and anti-hydroxyl radical activity were observed in aqueous B. edulis extract from the caps (half maximal inhibitory concentration [IC50] = 50.97 μg/ mL and 2.05 μg/mL, respectively), whereas the highest anti-nitric oxide radical activity was observed in aqueous B. edulis extract from the stipes (IC50 = 10.74 μg/mL). The ethanolic extract obtained from the mushroom stipe showed higher anti-superoxide anion radical activity (IC50 = 9.84 μg/mL) and ferric-reducing antioxidant power (22.14 mg ascorbic acid equivalents/g dry weight) compared with aqueous extracts. Total phenolic content for all extracts was similar but total flavonoid content was significantly higher in the aqueous B. edulis extract from the caps (4.5 mg quercetin equivalents/g dry weight). All crude extracts showed activity against the MCF-7 cell line, with the ethanolic extract of B. edulis prepared from stipes (IC50 = 56 μg/mL) being the most potent. This is, to our knowledge, the first report of the antiproliferative effects of crude aqueous and ethanolic extracts prepared from caps and stipes of wild-growing basidiocarps of B. edulis on the human breast MCF-7 cancer cell line.

The combination of polymer-based passive sampling to collect complex environmental mixtures of pollutants, the transfer of these mixtures into bioassays, and their related toxicological characterization is still in its infancy. However, this approach has considerable potential to improve environmental hazard and risk assessment for two reasons. First, the passive sampler collects a broad range of chemicals representing the fraction of compounds available for diffusion and (bio)uptake, excluding a large part of the matrix; thus, extensive sample cleanup which could discriminate certain compounds can be avoided. Second, the toxicological characterization of samples using bioassays is complementary to chemical (target) analysis within environmental monitoring because it captures all chemicals exerting the same mode of toxic action and acting jointly in mixtures, thus providing a comprehensive picture of their overall combined effects. The scientific literature describes a range of examples from the water phase where passive sampling is usually carried out in the kinetic uptake regime for most chemicals although some may already have reached equilibrium. The composition of the chemical mixture changes from the water phase to the passive sampling material because of kinetic effects and polymer/water partition coefficients which depend on the chemicals' hydrophobicity. In contrast, only a few applications in sediment and biota have been described, but amongst these some pioneering studies have demonstrated the feasibility and potential of this combined approach. This chapter gives an overview of what has been carried out in this research area, focusing on opportunities and challenges, and points out desirable future developments with a focus on the importance of choosing a suitable combination of sampling and dosing to transfer (or re-establish) the environmental mixture into the bioassay.