What Are the Sensory Attributes Associated with Consumer Acceptance of Yellow Oyster Mushrooms (Pleurotus citrinopileatus)?

The oyster mushroom is cultivated globally, renowned for its unique texture and umami flavor, as well as its rich content of nutrients and functional ingredients. This study aims to identify the descriptive sensory characteristics, assess the consumer acceptability of new superior lines and cultivars of yellow oyster mushrooms, in addition to exploring the relationship between these descriptive characteristics and consumer acceptability. Statistical analyses were performed using one-way analysis of variance (ANOVA), principal component analysis (PCA), and partial least squares regression (PLSR). Twenty attributes were delineated, including three related to appearance/color (gray, yellow, and white), four associated with the smell/odor of fresh mushroom (oyster mushroom, woody, fishy, and seafood smells), three pertaining to the smell/odor of cooked mushrooms (mushroom, umami, and savory smells), four describing flavor/taste (sweet, salty, umami, and savory tastes), and five for texture/mouthfeel (chewy, smooth, hard, squishy, and slippery textures). Consumer acceptability tests involved 100 consumers who evaluated overall liking, appearance, overall taste, sweetness, texture, savory taste, MSG taste, smell, color, purchase intention, and recommendation. The general oyster mushroom (548 samples) scored highest in acceptability. Seven attributes, namely fresh mushroom smell, seafood smell (fresh), fishy smell (fresh), umami smell (cooked), nutty smell (cooked), salty taste, and MSG taste with the exception of appearance showed significant differences among samples (p < 0.001). The three yellow oyster mushroom samples were strongly associated with attributes like hardness, softness (texture), sweet taste (745 samples), MSG taste, salty taste, squishy texture, and fishy smell (483 and 629 samples). The development of sensory lexicons and increasing consumer acceptance of new superior lines and cultivars of yellow oyster mushroom will likely enhance sensory quality and expand the consumer market, aligning with consumer needs and preferences.

Effects of Light on the Fruiting Body Color and Differentially Expressed Genes in Flammulina velutipes.

Light plays vital roles in fungal growth, development, reproduction, and pigmentation. In Flammulina velutipes, the color of the fruiting body exhibits distinct changes in response to light; however, the underlying molecular mechanisms remain unknown. Therefore, in this study, we aimed to analyze the F. velutipes transcriptome under red, green, and blue light-emitting diode (LED) lights to identify the key genes affecting the light response and fruiting body color in this fungus. Additionally, we conducted protein-protein interaction (PPI) network analysis of the previously reported fruiting body color-related gene, Fvpal1, to identify the hub genes. Phenotypic analysis revealed that fruiting bodies exposed to green and blue lights were darker than those untreated or exposed to red light, with the color intensifying more after 48 h of exposure to blue light compared to that after 24 h of exposure. Differentially expressed gene (DEG) analyses of all light treatments for 24 h revealed that the numbers of DEGs were 17, 74, and 257 under red, green, and blue lights, respectively. Subsequently, functional enrichment analysis was conducted of the DEGs identified under green and blue lights, which influenced the color of F. velutipes. In total, 103 of 168 downregulated DEGs under blue and green lights were included in the enrichment analysis. Among the DEGs enriched under both green and blue light treatments, four genes were related to monooxygenases, with three genes annotated as cytochrome P450s that are crucial for various metabolic processes in fungi. PPI network analysis of Fvpal1 revealed associations with 11 genes, among which the expression of one gene, pyridoxal-dependent decarboxylase, was upregulated in F. velutipes exposed to blue light. These findings contribute to our understanding of the molecular mechanisms involved in the fruiting body color changes in response to light and offer potential molecular markers for further exploration of light-mediated regulatory pathways.

Identification of two genes essential for basidiospore formation during the postmeiotic stages in Pleurotus ostreatus.

A sporeless strain is an important breeding target in the mushroom industry. However, basidiospore production in the oyster mushroom Pleurotus ostreatus has been shown to be impaired by single-gene mutations in only two meiosis-related genes, mer3 and msh4. This study proposed a strategy for identifying the genes essential for basidiospore formation after meiotic division to determine new targets for molecular breeding. RNA-seq analysis was performed to identify P. ostreatus genes that are specifically expressed in the gill tissue of fruiting bodies, where basidiospore formation occurs. Transcriptome data during fruiting development of Coprinopsis cinerea, in which the meiotic steps progress synchronously, were then used to identify genes that are active in the postmeiotic stages. Based on these comparative analyses, five P. ostreatus genes were identified. Plasmids containing expression cassettes for hygromycin B-resistance screening, Cas9, and single-guide RNA targeting each gene were introduced into the protoplasts of dikaryotic strain, PC9×#64, to generate dikaryotic gene disruptants. Among the obtained transformants, three dikaryotic pcl1 disruptants and two cro6c disruptants did not produce basidiospores. Microscopic analyses indicated that spore formation was arrested at particular stages in these gene disruptants. These results indicate that these two genes are essential for mature spore formation in this fungus.

Cochlear Implantation in the Elderly: Speech Performance, Associated Factor, Complication, and Surgical Safety.

BACKGROUND AND OBJECTIVES: The guidelines for cochlear implantation (CIs) are expanding, and the number of CI procedures performed on the elderly is increasing. The purpose of this study was to analyze the results and safety of cochlear implantation in the elderly, as well as to evaluate the predictive factors on CI outcomes. SUBJECTS AND METHODS: The study included 56 patients aged ≥40 years, who received CIs between 2009 and 2020. They were divided into two groups: 27 younger adults (40-64 years) and 29 elderly (>64 years). The study compared their pre- and postoperative speech perception and category of auditory performance (CAP) scores, surgical complications, and hospitalization periods. It also evaluated associated factors in the elderly group by examining categorical and continuous variables and postoperative CAP score. RESULTS: There was a significant improvement in speech recognition tests (both word and sentence) and CAP scores in both groups compared to the pre-implantation scores (p<0.001). Postoperative results were slightly lower in the elderly group than in younger adults for sentence recognition and CAP scores, except for word recognition. No significant associated factors were found on postoperative CAP scores, except for etiology. Postoperative CAP significantly improved in the sudden hearing loss group compared to the groups with other etiologies (p=0.045). The elderly group had more comorbidities than that in the younger adult group (p=0.026), but there were no significant differences in postoperative complications and hospitalization periods. CONCLUSIONS: While speech recognition and CAP scores were relatively lower in the elderly group compared to the younger adults, the elderly group showed significant improvements in audiological results after CI. Moreover, CI was safe and well tolerated in elderly patients.

Double-gene targeting with preassembled Cas9 ribonucleoprotein for safe genome editing in the edible mushroom Pleurotus ostreatus.

CRISPR/Cas9 has potential for efficient molecular breeding. Recently, a foreign-DNA-free gene-targeting technology was established by introducing a preassembled Cas9 ribonucleoprotein (RNP) complex into the oyster mushroom Pleurotus ostreatus. However, the target gene was restricted to such a gene like pyrG, since screening of a genome-edited strain was indispensable and could be performed via examination of 5-fluoroorotic acid (5-FOA) resistance caused by the disruption of the target gene. In this study, we simultaneously introduced the Cas9 RNP complex targeting fcy1, a mutation that conferred P. ostreatus resistance to 5-fluorocytosine (5-FC), together with that targeting pyrG. A total of 76 5-FOA resistant strains were isolated during the first screening. Subsequently, a 5-FC resistance examination was conducted, and three strains exhibited resistance. Genomic PCR experiments followed by DNA sequencing revealed that mutations were successfully introduced into fcy1 and pyrG in the three strains. The results indicated that double gene-edited mutants could be obtained in one experiment employing 5-FOA resistance screening for strains with Cas9 RNP incorporation. This work may pave the way for safe CRISPR/Cas9 technology to isolate mutant strains in any gene of interest without an ectopic marker gene.

Effect of prehydration solution on hearing threshold after chemotherapy in patients with head and neck cancers: a retrospective study.

BACKGROUND: The study aimed to evaluate the effect of prehydration solution on hearing thresholds after cisplatin chemotherapy. METHODS: In this retrospective cohort study, we reviewed the data of patients who underwent ≥3 courses of cisplatin-based chemotherapy for locally advanced head and neck cancers at a tertiary referral center (n=64). The dextrose solution (DW) group (n=26) received 2 L of normal saline and 1 L of 5% dextrose. The Hartmann solution (HS) group (n=38) received 2 L of normal saline and 1 L of HS. Hearing data were measured 1 day before starting the first course of chemotherapy, and again 20 days after the first, second, and third courses of chemotherapy. The severity of hearing loss was evaluated using the Common Terminology Criteria for Adverse Events (CTCAE). RESULTS: Thresholds at all frequencies after chemotherapy were greater in the DW group than in the HS group. The increase in thresholds in 1 to 4 kHz after the third course of chemotherapy was greater in the DW group than in the HS group. CTCAE grades after the second and third courses of chemotherapy were greater in the DW group than in the HS group. Logistic regression showed that the odds ratio for CTCAE grade 3 or 4 after the third course of chemotherapy in the DW group was 4.84 on univariate analysis. CONCLUSION: Prehydration using a solution with salt was associated with a decrease in change in hearing thresholds after cisplatin chemotherapy in patients with head and neck cancers.

Introducing multiple-gene mutations in Pleurotus ostreatus using a polycistronic tRNA and CRISPR guide RNA strategy.

The white-rot fungus Pleurotus ostreatus is an agaricomycete that is frequently used in molecular genetics studies as many useful tools are applicable to the fungus. In particular, efficient gene targeting using homologous recombination and CRISPR/Cas9 enables the introduction of a mutation in the gene of interest for functional analysis. Multiple genes encoding various lignocellulose-degrading enzymes are predicted to be present in the genome; therefore, analyses of multiple-gene mutants are required to elucidate the mechanisms underlying lignocellulose degradation by P. ostreatus. Conventional tools for generating multiple-gene mutations in P. ostreatus are laborious and time-consuming. Therefore, more efficient and practical methods are needed. In this study, we introduced CRISPR/Cas9-assisted multiple-gene mutations using a polycistronic tRNA and CRISPR guide RNA approach. The frequency (triple-gene mutation in fcy1, vp2, and 62347) was only 3.3% when a tetracistronic tRNA-sgRNA containing four different sgRNAs targeting fcy1, vp2, vp3, or 62347 was expressed. It increased to 20% (triple-gene mutation in vp1, vp2, and vp3) after a tricistronic tRNA-sgRNA was expressed with replaced/modulated promoter and tRNA sequences. This study demonstrated, for the first time, the applicability of a strategy to induce multiple-gene mutations in P. ostreatus in a transformation experiment.

Gene targeting of dikaryotic Pleurotus ostreatus nuclei using the CRISPR/Cas9 system.

Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)-assisted gene targeting is a promising method used in molecular breeding. We recently reported the successful introduction of this method in the monokaryotic Pleurotus ostreatus (oyster mushroom), PC9. However, considering their application in mushroom breeding, dikaryotic strains (with targeted gene mutations in both nuclei) need to be generated. This is laborious and time-consuming because a classical crossing technique is used. Herein, we report a technique that targets both nuclei of dikaryotic P. ostreatus, PC9×#64 in a transformation experiment using plasmid-based CRISPR/Cas9, with the aim of developing a method for efficient and rapid molecular breeding. As an example, we targeted strains with low basidiospore production ability through the meiosis-related genes mer3 or msh4. Four different plasmids containing expression cassettes for Cas9 and two different gRNAs targeting mer3 or msh4 were constructed and separately introduced into PC9×#64. Eight of the 38 dikaryotic transformants analyzed produced no basidiospores. Genomic PCR suggested that msh4 or mer3 mutations were introduced into both nuclei of seven out of eight strains. Thus, in this study, we demonstrated simultaneous gene targeting using our CRISPR/Cas9 system, which may be useful for the molecular breeding of cultivated agaricomycetes.

Association between Fat Distribution and Chronic Low Back Pain among 10,606 Adults: Data from the Korean National Health and Nutrition Examination Survey.

Obesity is associated with chronic low back pain (CLBP), but the association between fat distribution and CLBP is unclear. This cross-sectional study evaluated the relationship using the Korean National Health and Nutrition Examination Survey data. A total of 10,606 adults (average age: 45.4, female: 57.1%) were included. We estimated the regional fat distribution, waist circumference, and body fat proportion, compared the values in people with and without CLBP, and stratified the estimates by sex and obesity status using a multivariable linear model. There were no statistically significant differences in the average waist circumference between the people with and without CLBP (p = 0.731) and the average fat proportion between those with and without CLBP (p = 0.731). The average regional fat distribution was significantly higher in the people with CLBP than in those without CLBP, in the upper limbs (11.4%, 95% confidence interval [CI]: [11.3, 11.5] vs. 11.2%, 95% CI: [11.1, 11.3], p < 0.05) and in the lower limbs (31.9%, 95% CI: [31.6, 32.2] vs. 31.4%, 95% CI: [31.2, 31.6], p < 0.01). More obvious among men, fat distribution in the lower limbs is higher than in people without obesity (p < 0.001). People with CLBP tend to have a higher fat distribution in the limbs than those without it and obese people with CLBP would need to reduce the fat in the lower limbs.

Presence of Cholesterol in Non-Animal Organisms: Identification and Quantification of Cholesterol in Crude Seed Oil from Perilla frutescens and Dehydrated Pyropia tenera.

Studies have reported that cholesterol, a molecule found mainly in animals, is also present in some plants and algae. This study aimed to determine whether cholesterol exists in three dehydrated algae species, namely, Pyropia tenera, Saccharina japonica, and Undaria pinnatifida, and in one plant species, namely, Perilla frutescens (four perilla seed oil samples were analyzed). These species were chosen for investigation because they are common ingredients in East Asian cuisine. Gas chromatography-flame ionization detection (GC-FID) analysis found that cholesterol was present in P. tenera (14.6 mg/100 g) and in all four perilla seed oil samples (0.3-0.5 mg/100 g). High-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD) also demonstrated that cholesterol was present in P. tenera (14.2 mg/100 g) and allowed the separation of cholesterol from its isomer lathosterol. However, cholesterol could not be detected by HPLC-ELSD in the perilla seed oil samples, most likely because it is only present in trace amounts. Moreover, liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed the presence of cholesterol in both P. tenera and perilla seed oil. MRM results further suggested that lathosterol (a precursor of cholesterol) was present in P. tenera.

Interspecific hybridization between Ganoderma lingzhi and G. applanatum through protoplast fusion.

Interspecific hybridization between Ganoderma lingzhi and G. applanatum was attempted through polyethylene glycol (PEG) induced fusion technique. The protoplast isolation procedure was simplified, and we obtained a significant number of protoplasts from both Ganoderma species. The number of protoplasts obtained was 5.27 ± 0.31 × 107/mL in G. lingzhi and 5.57 ± 0.49 × 106/mL in G. applanatum. Osmotic stabilizer NaCl (0.4 M) at pH 5.8 and enzymolysis time 3.5 h have supported high frequency of protoplast regeneration. G. lingzhi and G. applanatum regeneration frequency was 1.73 ± 0.04% and 0.23 ± 0.02%, respectively. 40% of PEG induced high number of protoplast fusion the regeneration frequency was 0.09% on a minimal medium. Two hundred fifty-two fusant colonies were isolated from the following four individual experiments. Among them, ten fusants showed the mycelial morphological difference compared to their parents and other fusant isolates. The fruiting body could be generated on oak sawdust and wheat bran substrate, and a few of them showed recombined morphology of the parental strains. The highest yield and biological efficacy (BE) were recorded in GF248, while least in GF244. The hybridity of the fusant was established based on mycelia, fruiting morphology, and PCR fingerprinting. ISSR and RAPD profile analysis of ten fusants and parents depicted that fusants contained polymorphic bands, which specified the rearrangement and deletion of DNA in the fusants. A Dendrogram was constructed based on the RAPD profile, and the clustering data exhibited two major clusters: cluster I included the G. lingzhi and Cluster II, including the G. applanatum and fusant lines. Total polysaccharide (α, ß and total glucan) content was compared with fusants and parental strains. The present study highlighted the efficient methods for protoplast isolation from Ganoderma species. PEG-induced fusants showed high polymorphic frequency index, while the phenotypic characters showed high similarity to G. applanatum. A significant difference was observed in the mushroom yield and its total polysaccharide between the fusants and parental strains.

Bioactive Sterol Derivatives Isolated from the Pleurotus djamor var. Roseus Induced Apoptosis in Cancer Cell Lines.

OBJECTIVE: The aim of the present study is to isolate and characterize the bioactive compounds from Pleurotus djamor against human breast cancer (MDA-MD-231) and mouse T cell lymphoma (EL4) cell lines. MATERIALS AND METHODS: Sequential fractionization and column chromatography methods were involved in compound isolation. The structures of the isolated compound were determined by NMR, GC/MS, and X-ray crystallography studies. RESULTS: The isolated compounds 1- 4 [D-mannitol (C1), ergosta-5,7,22-trien-3ß-ol (C2), 5,8- epidioxy-ergosta-6-22-dien-3ß-ol (C3), and palmitic acid (C4)] are white crystal and amorphous powder in nature. All these compounds were isolated from this mushroom for the first time. In vitro lipid peroxidation activities of isolated compounds were determined by ferric thiocyanate (FTC) and thiobarbituric acid (TBA) method. The sterol derivatives C2 and C3 compounds displayed strong antioxidant activity and were not significantly different (p<0.05) to α-tocopherol. This finding elaborates on the isolation of a cytotoxic compound C2 and C3 from P. djamor via a rapid elution method. CONCLUSION: The compound C3 has exhibited better cytotoxic activity against MDA-MD-231 and EL4 cells. The present finding and data might provide new insights into the possible therapeutic and pharmaceutical use for the design of anti-cancer drugs from this edible mushroom.

Cultivation and Nutritional Value of Prominent Pleurotus spp.: An Overview.

Pleurotus species are commercially essential mushrooms and widely cultivated throughout the world. The production of Pleurotus mushrooms alone accounts for around 25% of that total cultivated mushrooms globally. In America and Europe, Pleurotus species are considered specialty mushrooms, whereas, in Korea, their cultivation is economically profitable, and it is one of the highly consumed species. Pleurotus species are predominantly found in tropical forests and often grow on fallen branches, dead and decaying tree stumps, and wet logs. Biographical studies have shown that the Pleurotus genus is among the more conspicuous fungi that induce wood decay in terrestrial ecosystems worldwide due to its formidable lignin-modifying enzymes, including laccase and versatile peroxidases. Pleurotus species can be grown easily due to their fast colonization nature on diversified agro-substrates and their biological efficiency 100%. Pleurotus mushrooms are rich in proteins, dietary fiber, essential amino acids, carbohydrates, water-soluble vitamins, and minerals. These mushrooms are abundant in functional bioactive molecules, though to influence health. Pleurotus mushrooms are finding unique applications as flavoring, aroma, and excellent preservation quality. Apart from its unique applications, Pleurotus mushrooms have a unique status delicacy with high nutritional and medicinal values. The present review provides an insight into the cultivation of Pleurotus spp. using different agro-waste as growth substances paying attention to their effects on the growth and chemical composition.

Anti-inflammatory effect of lovastatin is mediated via the modulation of NF-κB and inhibition of HDAC1 and the PI3K/Akt/mTOR pathway in RAW264.7 macrophages.

Lovastatin is a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor that is clinically used for the prevention of cardiovascular diseases. Although it has been reported that lovastatin has anti-inflammatory properties in several studies, how lovastatin regulates the inflammation is still unclear. To evaluate the effect of lovastatin on nitric oxide production (NO) in RAW264.7 macrophages, NO production assay was performed. Also, cell viability was measured to confirm cytotoxicity. Level of tumor necrosis factor-α (TNF-α) transcription was measured by reverse transcription polymerase chain reaction (RT-PCR) from total RNA in RAW264.7 cells. Western blot analysis and immunofluorescence staining were used to investigate the regulation of lovastatin on the expression, phosphorylation, and nuclear translocation of cellular proteins. The results of the present study revealed that lovastatin reduced nitric oxide production via the reduction of inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. The mRNA level of TNF-α was reduced in presence of lovastatin. In addition, lovastatin downregulated histone deacetylase 1 (HDAC1), resulting in the accumulation of acetylated histone H3 and heat shock protein 70. Furthermore, the expression of phosphoinositide 3-kinase catalytic subunits α and ß was reduced under lovastatin treatment, and the phosphorylation of Akt and mammalian target of rapamycin was consequently inhibited. Lovastatin also inhibited the phosphorylation of inhibitor of nuclear factor (NF)-κBα and the translocation of NF-κB into the nucleus. Therefore, the present study demonstrates that lovastatin inhibits the expression of pro-inflammatory mediators, including iNOS and TNF-α, through the suppression of HDAC1 expression, PI3K/Akt phosphorylation and NF-κB translocation in LPS-stimulated RAW264.7 macrophage cells.