Neurite outgrowth stimulatory effects of culinary-medicinal mushrooms and their toxicity assessment using differentiating Neuro-2a and embryonic fibroblast BALB/3T3

BACKGROUND: Mushrooms are not only regarded as gourmet cuisine but also as therapeutic agent to promote cognition health. However, little toxicological information is available regarding their safety. Therefore, the aim of this study was to screen selected ethno-pharmacologically important mushrooms for stimulatory effects on neurite outgrowth and to test for any cytotoxicity. METHODS: The stimulatory effect of mushrooms on neurite outgrowth was assessed in differentiating mouse neuroblastoma (N2a) cells. Neurite length was measured using Image-Pro Insight processor system. Neuritogenesis activity was further validated by fluorescence immunocytochemical staining of neurofilaments. In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively. RESULTS: Aqueous extracts of Ganoderma lucidum, Lignosus rhinocerotis, Pleurotus giganteus and Grifola frondosa; as well as an ethanol extract of Cordyceps militaris significantly (p < 0.05) promoted the neurite outgrowth in N2a cells by 38.4 ± 4.2%, 38.1 ± 2.6%, 33.4 ± 4.6%, 33.7 ± 1.5%, and 35.8 ± 3.4%; respectively. The IC(50) values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts. CONCLUSION: Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health

Giant oyster mushroom, Pleurotus giganteus (Agaricomycetes): Current status of the cultivation methods, chemical composition, biological, and health-promoting properties

Pleurotus giganteus (Berk.) Karunarathna & Hyde is one of the largest edible mushrooms which can be found either in groups or solitary and in soil and buried woods, but seldom in grassland. It has been recorded in Southeast Asia, Australasia, and China by either its common names or local names. Previously known as Lentinus/Panus giganteus, its unique physical characteristics and the utilization of molecular tools have since anchored its taxonomical position in the Pleurotus genus. A sawdust-based substrate coupled with soil casing method is described as a standard cultivation procedure for P. giganteus. The basidiocarp of P. giganteus is found to be rich in carbohydrates, proteins, polyunsaturated fatty acids, and polysaccharides. Both ethanol and aqueous extracts of P. giganteus have been studied in vitro for its antioxidative, antifungal, anticancer, hepatoprotective, and neurite outgrowth capabilities. The review covers the optimum cultivation methods of P. giganteus, as well as its various biological activities and medicinal properties that have been studied by different approaches. Available data in regards to the chemical compounds present in P. giganteus are also compiled for future references. Overall, both in vitro and in vivo studies have shown significant bioactivity comparable to other recognized Pleurotoid mushrooms

Gastroprotective Effects of Lion’s Mane Mushroom Hericium erinaceus

Hericium erinaceus is a famous tonic in oriental medicine. The gastroprotective effects of aqueous extract of H. erinaceus against ethanol-induced ulcers in Sprague Dawley rats were investigated. The possible involvements of lipid peroxidation, superoxide dismutase, and catalase were also investigated. Acute toxicity study was performed. The effects of aqueous extract of H. erinaceus on the ulcer areas, ulcer inhibition, gastric wall mucus, gross and histological gastric lesions, antioxidant levels, and malondialdehyde (MDA) contents were evaluated in ethanol-induced ulcer in vivo. In acute toxicity study, a high dose of 5 g/kg did not manifest any toxicological signs in rats. The extract promoted ulcer protection as ascertained by a significant reduction of the ulcer area. Furthermore, it exhibited a significant protection activity against gastric mucosal injury by preventing the depletion of antioxidant enzymes. The level of MDA was also limited in rat stomach tissues when compared with the ulcer control group. Immunohistochemistry showed upregulation of HSP70 protein and downregulation of BAX protein in rats pretreated with the extract. The aqueous extract of H. erinaceus protected gastric mucosa in our in vivo model. It is speculated that the bioactive compounds present in the extract may play a major role in gastroprotective activity

Intrastrain Comparison of the Chemical Composition and Antioxidant Activity of an Edible Mushroom, Pleurotus giganteus

Two strains of Pleurotus giganteus (commercial and wild) were tested for their ability to induce neurite outgrowth in rat pheochromocytoma (PC12) and mouse neuroblastoma-2a (N2a) cells. Treatment with the mushroom extracts resulted in neuronal differentiation and neuronal elongation, but not nerve growth factor (NGF) production. Linoleic acid (4.5–5.0%, w/w) which is a major fatty acid present in the ethanol extract promoted NGF biosynthesis when augmented with low concentration of NGF (5 ng/mL). The two strains of mushroom were found to be high in protein (154–192 g kg−1), total polysaccharides, phenolics, and flavonoids as well as vitamins B1, B2, and B3. The total phenolics present in the mushroom extracts were positively correlated to the antioxidant activity (free radical scavenging, ferric reducing power, and lipid peroxidation inhibition). To conclude, P. giganteus could potentially be used in well-balanced diet and as a source of dietary antioxidant to promote neuronal health

Bibliometric Analysis of Mushroom Poisoning: From Diversity to Clinical Management

The earliest publication related to mushroom poisoning dates back to 1837. To date, bibliometric analysis related to the field of mushroom poisoning has not been published. This study aimed to assess the most significant publications in this field as well as the associated trends and important drivers in the research related to mushroom poisoning. The Scopus database was screened to identify relevant publications on mushroom poisoning. A total of 985 publications with a minimum of five citations were identified and analyzed. Pearson’s correlation demonstrated an insignificant weak negative correlation (Pearson’s correlation of −0.020, P > 0.01) between the number of years since publication and the number of citation counts of a paper. Bradford’s law of scattering revealed that one-third of publications were published in 31 core journals, with Clinical Toxicology topping the list (41 papers). VOSviewer was used to generate a network visualization based on country. The United States was the largest contributor of publications on mushroom poisoning, contributing 19.6% of 985. China is an emerging leader in publications on mushroom poisoning research since 2011, with the most recent average publication year of 2011.18. A term map was also created to visualize the co-occurrence of key terms, whereby Amanita phalloides–related research appeared to be the most frequently published topic in this field. In conclusion, the results of this bibliometric study shed light on the status of mushroom poisoning research and can guide investigators on current research trends for high-impact knowledge contribution in the field

The Changing Landscape for Stroke\ua0Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non\u2013vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase&nbsp;1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients\u2019 baseline characteristics. Atrial fibrillation&nbsp;disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age&nbsp; 6575 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score&nbsp; 652; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc&nbsp;= 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N&nbsp;= 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in&nbsp;Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in&nbsp;Asia&nbsp;and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701

Neurite outgrowth stimulatory activity of an edible mushroom Pleurotus Giganteus in differentiating neuroblastoma-2a cells / Phan Chia Wei

synaptic connections is an important process for the establishment of synaptic connections during development, as well as neuronal regeneration in neuropathological conditions or injury. With growing concerns over neurodegenerative diseases attributed to impairment of neurite outgrowth e.g. dementia and Alzheimer’s disease, identification of alternative therapeutics has become paramount. One way to prevent and/or delay the onset of such diseases is by discovering alternative therapeutic molecules from functional foods. One such candidate is the edible mushroom (higher Basidiomycetes). In this study, eight culinary-medicinal mushrooms were evaluated for neurite outgrowth stimulatory effects by using neuroblastoma-2a (N2a) cells as an in vitro model. The mushroom extracts were also subjected to in vitro neuro- and embryotoxicity tests using N2a and 3T3 fibroblasts cell lines. The preliminary results showed that the aqueous extract of Pleurotus giganteus significantly (p < 0.05) promoted neurite outgrowth in N2a cells by 33.4 ± 4.6%. The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 hours exposure of N2a and 3T3 cells to the mushroom extract. The basidiocarps of P. giganteus were then analysed for various nutritional attributes. The mushroom composed of protein (15.4–19.2 g/100 g), polysaccharides, phenolics, and flavonoids as well as vitamins B1, B2, and B3. The antioxidant properties of the aqueous and ethanol extracts of P. giganteus were investigated. The results indicated that the aqueous extract of P. giganteus exhibited scavenging of 2,2-diphenyl-1-picrylhyd-razyl (DPPH) radical with an IC50 value of 21.46 ± 6.95 mg/mL. Based on the ferric reducing antioxidant power (FRAP) assay, the reducing power of the mushroom extracts was in the range of 1.17–3.88 μM FeSO·7H2O/g mushroom and the ethanol extract showed lipid peroxidation inhibitory activity of 49.58–49.80%. The efficacy of the chemical constituents of P. giganteus (linoleic acid, oleic acid, cinnamic acid, iv caffeic acid, p-coumaric acid, succinic acid, benzoic acid, and uridine) for neurite outgrowth activity was investigated. Uridine (100 μM) increased the number of neurite bearing cells by 43.1 ± 0.5%, which was about 1.8-fold higher than NGF (50 ng/mL)- treated cells. In this study, we demonstrated that uridine of P. giganteus (1.80 ± 0.03 g/100g mushroom extract) increased the phosphorylation of extracellular-signal regulated kinases (ERKs) and protein kinase B (Akt); simultaneously promoting neurite outgrowth in N2a cells. Neurite outgrowth stimulatory activity was inhibited by the inactivation of mitogen-activated protein kinase (MEK)/ERKs and Akt signaling with specific inhibitors. Further, phosphorylation of the mammalian target of rapamycin (mTOR) was also increased. MEK/ERK and PI3K-Akt-mTOR further induced phosphorylation of cAMP-response element binding protein (CREB) and expression of growth associated protein 43 (GAP43), tubulin alpha 4a (TUBA4A), and tubulin beta 1 (TUBb1); all of which promoted neurite outgrowth of N2a cells. In conclusion, these findings demonstrated that P. giganteus may enhance neurite outgrowth and one of the key bioactive molecule responsible for neurite outgrowth is uridine

A Status Review of the Bioactive Activities of Tiger Milk Mushroom Lignosus rhinocerotis (Cooke) Ryvarden

Edible and medicinal mushrooms are regularly used in natural medicines and home remedies since antiquity for ailments like fever, inflammation, and respiratory disorders. Lignosus rhinocerotis (Cooke) Ryvarden is a polypore found in Malaysia and other regions in South East Asia. It can be located on a spot where a tigress drips milk while feeding, hence the name “tiger's milk mushroom.” The sclerotium of L. rhinocerotis is highly sought after by the native communities in Malaysia to stave off hunger, relieve cough and asthma, and provide stamina. The genomic features of L. rhinocerotis have been described. The pharmacological and toxicity effects, if any, of L. rhinocerotis sclerotium have been scientifically verified in recent years. In this review, the validated investigations including the cognitive function, neuroprotection, immune modulation, anti-asthmatic, anti-coagulation, anti-inflammatory, anti-microbial/ anti-viral, anti-obesity, anti-cancer/ anti-tumor, and antioxidant properties are highlighted. These findings suggest that L. rhinocerotis can be considered as an alternative and natural medicine in the management of non-communicable diseases. However, there is a paucity of validation studies including human clinical trials of the mycochemicals of L. rhinocerotis

Synthesized 2-Trifluoromethylquinazolines and Quinazolinones Protect BV2 and N2a Cells against LPS- and H2O2-induced Cytotoxicity

BACKGROUND: Microglia are associated with neuroinflammation, which plays a key role in the pathogenesis of neurodegenerative diseases. It has been reported that some quinazolines and quinazolinones possess anti-inflammatory properties. However, the pharmacological properties of certain quinazoline derivatives are still unknown. OBJECTIVE: The antioxidant, cytotoxic, and protective effects of a series of synthesized 2-trifluoromethylquinazolines (2, 4, and 5) and quinazolinones (6-8) in lipopolysaccharide (LPS)-murine microglia (BV2) and hydrogen peroxide (H2O2)-mouse neuroblastoma-2a (N2a) cells were investigated. METHOD: The antioxidant activity of synthesized compounds was evaluated with ABTS and DPPH assays. The cytotoxic activities were determined by MTS assay in BV2 and N2a cells. The production of nitric oxide (NO) in LPS-induced BV2 microglia cells was quantified. RESULTS: The highest ABTS and DPPH scavenging activities were observed for compound 8 with 87.7% of ABTS scavenge percentage and 54.2% DPPH inhibition. All compounds were non-cytotoxic in BV2 and N2a cells at 5 and 50 μg/mL. The compounds which showed the highest protective effects in LPS-induced BV2 and H2O2-induced N2a cells were 5 and 7. All tested compounds, except 4, also reduced NO production at concentrations of 50 μg/mL. The quinazolinone series 6-8 exhibited the highest percentage of NO reduction, ranging from 38 to 60%. Compounds 5 and 8 possess balanced antioxidant and protective properties against LPS- and H2O2-induced cell death, thus showing great potential to be developed into anti-inflammatory and neuroprotective agents. CONCLUSION: Compounds 5 and 7 were able to protect the BV2 and N2a cells against LPS and H2O2 toxicity, respectively, at a low concentration (5 μg/mL). Compounds 6-8 showed potent reduction of NO production in BV2 cells