Prevalence and risk factors for chronic kidney disease among older adult patients with schizophrenia in Taiwan.

OBJECTIVES: There is an increasing incidence and prevalence of patients with chronic kidney disease (CKD) worldwide. Little is known the prevalence of CKD among older patients with schizophrenia. The purpose of this study was to investigate the prevalence of CKD and its risk factors in older adults with schizophrenia. METHODS: In this cross-sectional study, a convenience sample of 240 patients with schizophrenia age 50 or older were recruited. In addition to demographic and clinical data, participants' estimated glomerular filtration rate was calculated using the Modification of Diet in Renal Disease equation based on age, sex, ethnicity, and serum creatinine level determined from a blood sample taken from participants. RESULTS: The overall prevalence of CKD was 11.3%. Those with CKD group were older, had a longer duration of psychiatric illness, a higher body mass index (BMI), and diagnoses of hypertension compared to those in the non-CKD group. Independent of other risk factors, older age and BMI were significantly associated with CKD. CONCLUSIONS: This study found that the overall prevalence of CKD in older patients with schizophrenia was 11.3%. Risk factors for CKD in this population were older age and higher BMI. In addition to early identification and early treatment of CKD in older patients with schizophrenia, clinicians should actively manage the risk factors identified in this study, such as higher BMI and older age.

Behavioral and Neural Assessments of Auditory Skill Development After Hearing Instrument Fitting in Children: Case Reports and Clinical Implications.

PURPOSE: The purpose of this study was to comprehensively monitor the auditory skill development of children with hearing loss after hearing instrument fitting, and a battery of four assessments was proposed. METHOD: This battery was designed to fill the gap in speech discrimination in clinically available evaluations. The battery includes both behavioral and neural assessments. On the other hand, both tests in structured settings (sound-treated booth) and daily life were included in the battery. The four assessments include visual reinforced infant speech discrimination (VRISD), cortical auditory evoked potentials (CAEP), Auditory Skills Checklist (ASC), and Parents' Evaluation of Aural/Oral Performance of Children (PEACH). RESULTS: Two cases were reported, and their clinical implications were discussed. CONCLUSIONS: The proposed comprehensive assessment battery is suitable for evaluating children who are developmentally appropriate for visual reinforcement audiometry. More importantly, the VRISD assessment fills in the current gap, which is the discrimination stage, for the available clinical tests for assessing auditory developmental stages.

Innate-like bystander-activated CD38+ HLA-DR+ CD8+ T cells play a pathogenic role in patients with chronic hepatitis C.

BACKGROUND AND AIMS: HCV-specific T cells are few and exhausted in patients with chronic hepatitis C (CHC). Whether these T cells are responsible for the liver damage and fibrosis is still debated. However, cluster of differentiation 38-positive (CD38+ ) human leukocyte antigen DR-positive (HLA-DR+ ) CD8+ T cells are regarded as bystander CD8+ T cells that cause liver injury in acute hepatitis. We propose that these innate CD8+ T cells play a pathogenic role in CHC. METHODS: Lymphocytes from peripheral blood were obtained from 108 patients with CHC and 43 healthy subjects. Immunophenotyping, functional assays, T-cell receptor (TCR) repertoire, and cytotoxic assay of CD38+ HLA-DR+ CD8+ T cells were studied. RESULTS: The percentage of CD38+ HLA-DR+ CD8+ T cells increased significantly in patients with CHC. These cells expressed higher levels of effector memory and proinflammatory chemokine molecules and showed higher interferon-γ production than CD38- HLA-DR- CD8 T cells. They were largely composed of non-HCV-specific CD8+ T cells as assessed by HLA-A2-restricted pentamers and next-generation sequencing analysis of the TCR repertoire. In addition, these CD38+ HLA-DR+ CD8+ T cells had strong cytotoxicity, which could be inhibited by anti-DNAX accessory molecule 1, anti-NKG2 family member D, and anti-natural killer NKp30 antibodies. Lastly, the percentage of CD38+ HLA-DR+ CD8+ T cells was significantly associated with liver injury and fibrosis and decreased significantly along with serum alanine aminotransferase normalization after successful direct-acting antiviral treatment. CONCLUSIONS: The TCR-independent, cytokine-responsive bystander CD38+ HLA-DR+ CD8+ T cells are strongly cytotoxic and play a pathogenic role in patients with CHC.

Developing new scales for assessing parents' aural and oral rehabilitation skills to interact with children with hearing loss.

OBJECTIVE: Scales for evaluating the teaching and behavioural skills of parents enrolled in aural and oral rehabilitation programs for children with hearing loss are lacking. This study developed and validated scales for assessing parental teaching and behavioural skills of those parents for use in guiding their child to develop language and communication skills. DESIGN: Scales were constructed and evaluated using exploratory and confirmatory factor analysis. The performance of parents in teaching and behavioural skills was also explored. STUDY SAMPLE: The teaching and behavioural skills of 344 parents (179 for scale development and 165 for validation) were rated by their intervention therapists using parental teaching skill (PTS) and parental behavioural skill (PBS) scales. RESULTS: Good reliability and model fit (validity) were observed for both scales, which ultimately included 13 and 10 items, respectively. Maternal educational level was a significant indicator of their performance ratings. CONCLUSIONS: The PTS and PBS scales were validated and can be used by early intervention professionals to evaluate the relative interaction and behavioural skills of parents of children with hearing loss, and enrolled in listening and spoken language intervention programs.

Exposure to air pollution and scarlet fever resurgence in China: a six-year surveillance study.

Scarlet fever has resurged in China starting in 2011, and the environment is one of the potential reasons. Nationwide data on 655,039 scarlet fever cases and six air pollutants were retrieved. Exposure risks were evaluated by multivariate distributed lag nonlinear models and a meta-regression model. We show that the average incidence in 2011-2018 was twice that in 2004-2010 [RR = 2.30 (4.40 vs. 1.91), 95% CI: 2.29-2.31; p < 0.001] and generally lower in the summer and winter holiday (p = 0.005). A low to moderate correlation was seen between scarlet fever and monthly NO2 (r = 0.21) and O3 (r = 0.11). A 10 µg/m3 increase of NO2 and O3 was significantly associated with scarlet fever, with a cumulative RR of 1.06 (95% CI: 1.02-1.10) and 1.04 (95% CI: 1.01-1.07), respectively, at a lag of 0 to 15 months. In conclusion, long-term exposure to ambient NO2 and O3 may be associated with an increased risk of scarlet fever incidence, but direct causality is not established.

Oral intake of Streptococcus thermophil us improves knee osteoarthritis degeneration: A randomized, double-blind, placebo-controlled clinical study.

This preliminary clinical study demonstrates the possibility of a new species of probiotic for improvement of the degeneration of knee osteoarthritis (KOA). TCI633 (Streptococcus thermophil us) is a newly founded bacterium from human breast milk, and it is able to produce hyaluronate (HA) in gastrointestinal (GI) tract. A recent study has proved that TCI633 can substantially alleviate synovial tissue inflammation and cartilage damage in the animal models, but so far it has never been applied in clinical intervention. In this study, we recruited 80 subjects and conducted 12 weeks clinical trial to validate the efficacy of TCI633 for improvement of the progression of KOA. TCI633 could improve serum collagen type II C-telopeptide (sCTX-II) and serum C-reactive protein (sCRP) by 41.58% and 39.58%, respectively, after the study. The improvement rates for sCTX-II and sCRP in TCI633 group were 54% and 57%, respectively, at 12 weeks. Compared to the results of placebo, the indistinct improvement progresses of sCTX-II and sCRP might be caused by the uneventful distribution of K/L populations between the TCI633 and placebo groups, a short term of study period, and few recruited subjects. Moreover, the results of Western Ontario and McMaster Universities (WOMAC) questionnaires show that TCI633 might retard the progression and development of KOA after the trial. In brief, this preliminary research may provide an alternative approach to the improvement of KOA by probiotics although more detailed investigations should be conducted for solid conclusions.

Acute kidney injury and the risk of mortality in patients with methanol intoxication.

BACKGROUND: Methanol poisoning is a serious public health issue in developing countries, but few data are available in the literature on acute kidney injury (AKI) after methanol intoxication. METHODS: This study examined the clinical features, spectrum and outcomes of AKI in patients with methanol intoxication and evaluated the predictors of mortality after methanol intoxication. A total of 50 patients with methanol intoxication were seen at Chang Gung Memorial Hospital between 2000 and 2013. Patients were grouped according to the status of renal damage as AKI (n = 33) or non-AKI (n = 19). Demographic, clinical, laboratory, and mortality data were obtained for analysis. RESULTS: Most patients were middle-aged (47.8 ± 14.9 years), predominantly male (74.0%), and habitual alcohol consumers (70.0%). Most incidents were oral exposures (96.0%) and unintentional (66.0%). Two (4.0%) patients attempted suicide by intravenous injection of methanol. Five (10.0%) patients suffered methanol intoxication after ingestion of methomyl pesticide that contained methanol as a solvent. Compared to non-AKI patients, the AKI patients were older (50.9 ± 13.7 versus 41.6 ± 15.6 years, P = 0.034), predominantly male (90.9% versus 42.8%, P = 0.000), more habitual alcohol users (84.8% versus 41.2%, P = 0.001) and had more unintentional exposures (82.8% versus 35.3%, P = 0.001). Furthermore, there was a higher incidence of respiratory failure (63.6% versus 29.4%, P = 0.022) in the AKI group than in the non-AKI group, respectively. The laboratory studies revealed that the AKI patients suffered from more severe metabolic acidosis than the non-AKI patients. By the end of this study, 13 (39.5%) AKI patients and 1 (5.9%) non-AKI patient had died. The overall in-hospital hospital mortality rate was 28%. In a multivariate binary logistic regression model, it was demonstrated that AKI (odds ratio 19.670, confidence interval 1.026-377.008, P = 0.048) and Glasgow coma scale score (odds ratio 1.370, confidence interval 1.079-1.739, P = 0.010) were significant factors associated with mortality. The Kaplan-Meier analysis disclosed that AKI patients suffered lower cumulative survival than non-AKI patients (log-rank test, chi-square = 5.115, P = 0.024). CONCLUSIONS: AKI was common (66.0%) after methanol intoxication and was predictive of in-hospital hospital mortality. The development of AKI was associated with a 19.670-fold higher risk of in-hospital mortality.

Influences of Conjugation Length on Organic Field-Effect Transistor Performances and Thin Film Structures of Diketopyrrolopyrrole-Oligomers.

Here, two diketopyrrolopyrrole (DPP)-based oligomers, DPP-4T and DPP-6T, are studied to reveal the influences of conjugation length on thin-film morphology and organic field-effect transistor (OFET) performances. PDMS-assisted crystallization in a solvent-annealing chamber is applied to prepare crystal arrays of DPP-4T and DPP-6T to optimize the quality of charge channels for OFET characterizations. To deliver insights into microstructure and morphology of thin films, a characterization procedure for determining molecular packing in thin film and crystallinity of the crystal arrays is presented via grazing incidence wide-angle X-ray scattering, electron diffraction, and lattice simulation software package (Cerius2). With the lattice parameters derived from analyses of grazing incidence wide-angle X-ray scattering (GIWAXS) and electron diffraction (ED), the lattice modeling results indicate that the inferior organic field-effect transistor (OFET) performances of DPP-6T are attributed to longer π-stacking distance. Also, less-ordered molecular arrangement and lower continuity of crystalline domains, both of which are revealed from crystallinity results, lead to lower mobility of DPP-6T. In this case, longer conjugated backbones with more conformational degrees of freedom thus cause inherent crystal defects during the crystal growth process, despite the potential to enhance intermolecular π-orbital overlap. Therefore, to achieve better OFET performance, suitable backbone length makes conjugated oligomers give high intermolecular π-orbital overlap and low density of structural disorder, which are the priorities for constructing good charge channel.

Economic Conditions May Contribute to Increased Violence toward Children: A Nationwide Population-Based Analysis of Pediatric Injuries in Taiwanese Emergency Departments.

Childhood injuries are unfortunately common. Analysis procedures may assist professionals who work with children with developing preventive measures for protecting children's wellness. This study explores the causes of pediatric injuries presenting to an emergency department in Taiwan. This nationwide, population-based study was conducted using data from the National Health Insurance Research Database of Taiwan (NHIRD). Patients aged <18 years were identified from approximately one million individuals listed in the NHIRD. We followed up with these patients for nine years and analyzed the causes of injuries requiring presentation to an emergency department. Variables of interest were age, sex, injury mechanisms, and temporal trends. A total of 274,028 children were identified in our study. Between 2001 and 2009, the leading causes of pediatric injuries treated in emergency departments were motor vehicle injuries, falls, and homicide. The overall incidence of injuries declined over the course of the study because of reductions in motor vehicle accidents and falls. The incidence of homicide increased during the study period, particularly between 2007 and 2009. A moderately inverse correlation between homicide rate and economic growth was observed (correlation coefficient: -0.613, p = 0.041). There was a general decline in pediatric injuries between 2001 and 2009. Public policy changes, including motorcycle helmet laws and increases in alcohol taxes, may have contributed to this decline. Unfortunately, the incidence of homicide increased over the course of the study. Ongoing financial crises may have contributed to this increase. Multidisciplinary efforts are required to reduce homicide and reinforce the importance of measures that protect children against violence.

Current and Future Research Trends in Agricultural and Biomedical Applications of Medicinal Mushrooms and Mushroom Products (Review).

Here, we analyze the status and future trends in the study of medicinal mushrooms (MMs) in agriculture and biomedicine. Fruiting bodies of mushrooms are heterotrophic organisms that obtain all their nutritive requirements from the substrate. Mushroom substrates are agro-industrial plant residues and forest byproducts, which are usually called lignocellulosic materials. A good substrate for mushroom growth must be suitable both chemically and physically, as well as have the proper conditions for microbial activities. Under suitable conditions, mushrooms can excrete key enzymes for unlocking indigestible lignocellulosic biomasses to help provide sources of nourishment. The production of enzymes by mushroom mycelia plays a crucial part in the colonization process and is an important determinant of mushroom yields. The sense of purpose and vision for mushroom industries is also briefly discussed. Special attention is given to the use of mushroom extracts with antiphytopathologenic and insecticidal properties in modern agriculture. In the second part of this article, we summarize biomedical applications of medicinal mushrooms, which are currently used as 1) dietary food, 2) dietary supplement products, 3) a new class of drugs called "mushroom pharmaceuticals or mushroom drugs", 4) natural biocontrol agents in plant protection demonstrating insecticidal, fungicidal, bactericidal, herbicidal, nematocidal, and antiphytoviral activities, and 5) cosmeceuticals. We also aimed to draw attention to many critically important unsolved problems in the future development of medicinal mushroom science in the 21st century, including problems of production, standardization, and safety of mushroom dietary supplement products, as well as to discuss the problems of developing new medicinal mushroom drugs based not only on beta-glucans polysaccharides but also on low-molecular-weight secondary metabolites.

Increased risk of pernicious anemia following scabies: a nationwide population-based matched-cohort study.

OBJECTIVES: Scabies is a common and annoying disorder. Pernicious anemia (PA) is a serious disease which, when untreated, leads to death. Mounting evidence suggests that immune-mediated inflammatory processes play a role in the pathophysiology of both diseases. The relationship between these two diseases has not been investigated. We conducted this study to explore the potential relationship between scabies and PA. MATERIALS AND METHODS: This nationwide, population-based study was conducted using the National Health Insurance Research Database of Taiwan. In total, 5,407 patients with scabies were identified as a study group and 20,089 matched patients were randomly selected as a control group. We tracked patients in both groups for a 7-year period to identify the incidence of PA. The demographic characteristics and comorbidities of the patients were analyzed, and Cox proportional hazards regression was used to calculate the hazard ratios for PA. RESULTS: Of the 25,496 patients in this study, 183 (0.7%) patients with newly diagnosed PA were identified during the 7-year follow-up period; 71 of 5,407 (1.3%) from the scabies group and 112 of 20,089 (0.6%) from the control group. Patients with scabies had a higher risk of subsequent PA, with a crude hazard ratio of 2.368. After adjusting for covariates, the adjusted hazard ratio was 1.51 (95% confidence interval: 1.09-2.08). CONCLUSION: This study demonstrated an increased risk of PA (adjusted hazard ratio 1.51) among patients with scabies. Immune-mediated inflammatory processes may contribute to this association. Further studies are warranted to investigate the entire pathological mechanisms between these two diseases. Physicians should pay attention to patients with history of scabies presented with anemia. Further confirmative tests of PA may contribute to correct diagnosis and initiation of vitamin B12 supplement.

Increase the risk of intellectual disability in children with scabies: A nationwide population-based cohort study.

Scabies is a common and distressing disease caused by the mite Sarcoptes scabiei var. hominis. Psychiatric disorder in childhood is an important disease and easily neglected. There are several similarities in scabies and psychiatric disorders in childhood (PDC). Both of them may present with pruritus. They are relatively common in patients with lower socioeconomic status and crowded environment. Furthermore, immune-mediated inflammatory processes play a role in the pathophysiology in both diseases. An association between scabies and psychiatric disorders may exist. This nationwide population-based cohort study utilized data from the National Health Insurance Research Database to investigate the relationship between scabies and PDC. A total of 2137 children with scabies were identified as the study group and 8548 age- and sex-matched children were selected as the control group. A total of 607 (5.68%) children developed PDC during the 7-year follow-up period. The overall incidences of PDC are similar but patients with scabies had a higher risk of developing intellectual disability (ID) (scabies group vs control group: 1.3% vs 0.6%, adjusted hazard ratio: 2.04 and 95% confidence interval: 1.25-3.32). The immune-mediated inflammatory processes of both diseases were reviewed and may contribute to the 104% increased risk of interleukin in patients with scabies. We suggest a more comprehensive management in treating patients with scabies or ID. Early and comprehensive treatment of scabies and other risk factors may decrease the risk of subsequent ID. When we approach patients with ID, concurrent evaluation of scabies and other risk factors may contribute to successful management.

Cofacial Versus Coplanar Arrangement in Centrosymmetric Packing Dimers of Dipolar Small Molecules: Structural Effects on the Crystallization Behaviors and Optoelectronic Characteristics.

Two D-π-A-A molecules (MIDTP and TIDTP) composed of an electron-rich ditolylamino group (D) and an electron-deficient 5-dicyanovinylenylpyrimidine (A-A) fragment bridged together with indeno[1,2-b]thiophene (IDT) were synthesized. These molecules provide an opportunity to examine in-depth the impact of side-chain variations (methyl vs p-tolyl) on the crystallization behaviors, solid-state morphology, physical properties, and optoelectronic characteristics relevant for practical applications. X-ray analyses on single-crystal structures indicate that methyl-substituted MIDTP forms "coplanar antiparallel dimers" via C-H···S interactions and organizes into an ordered slip-staircase arrays. In contrast, p-tolyl-bearing TIDTP shows "cofacial centrosymmetric dimers" via π-π interactions and packs into a less-ordered layered structures. The X-ray diffraction analyses upon thermal treatment are consistent with a superior crystallinity of MIDTP, as compared to that of TIDTP. This difference indicates a greater propensity to organization by introduction of the smaller methyl group versus the bulkier p-tolyl group. The increased propensity for order by MIDTP facilitates the crystallization of MIDTP in both solution-processed and vacuum-deposited thin films. MIDTP forms solution-processed single-crystal arrays that deliver OFET hole mobility of 6.56 × 10(-4) cm(2) V(-1) s(-1), whereas TIDTP only forms amorhpous films that gave lower hole mobility of 1.34 × 10(-5) cm(2) V(-1) s(-1). MIDTP and TIDTP were utilized to serve as donors together with C70 as acceptor in the fabrication of small-molecule organic solar cells (SMOSCs) with planar heterojunction (PHJ) or planar-mixed heterojunction (PMHJ) device architectures. OPV devices based on higher crystalline MIDTP delivered power conversion efficiencies (PCEs) of 2.5% and 4.3% for PHJ and PMHJ device, respectively, which are higher than those of TIDTP-based cells. The improved PCEs of MIDTP-based devices are attributed to better hole-transport character.

Flat-on ambipolar triphenylamine/C60 nano-stacks formed from the self-organization of a pyramid-sphere-shaped amphiphile.

A giant amphiphile, which is constructed with an amorphous nano-pyramid (triphenylamine, TPA) and a crystalline nano-sphere (C60), was synthesized. Structural characterization indicates that this pyramid-sphere-shaped amphiphile (TPA-C60 ) forms a solvent-induced ordered phase, in which the two constituent units self-assemble into alternating stacks of two-dimensional (2D) TPA and C60 nano-sheets. Due to the complexity of the molecular structure and the amorphous nature of the nano-pyramid, phase formation was driven by intermolecular C60-C60 interactions and the ordered phase could not be reformed from the TPA-C60 melt. Oriented crystal arrays of TPA-C60 , which contain flat-on TPA/C60 nano-stacks, can be obtained via a PDMS-assisted crystallization (PAC) technique. The flat-on dual-channel supramolecular structure of TPA-C60 delivered ambipolar and balanced charge-transport characteristics with an average µe of 2.11 × 10-4 cm2 V-1 s-1 and µh of 3.37 × 10-4 cm2 V-1 s-1. The anisotropic charge-transport ability of the pyramid-sphere-shaped amphiphile was further understood based on the lattice structure and the lattice orientation of TPA-C60 revealed from electron diffraction analyses.

A Facile PDMS-Assisted Crystallization for the Crystal-Engineering of C60 Single-Crystal Organic Field-Effect Transistors.

Poly(dimethylsiloxane) (PDMS)-assisted crystallization (PAC) is a facile method to produce oriented C60 crystal arrays. Changing the drying mechanism from evaporation to solvent absorption (by PDMS) widens the solvent selection and facilitates the engineering of both the macroscopic shape and the microscopic lattice structure of the crystal arrays. The method also shows the potential to be applied to other organic semiconductors and large-area production.

Mushrooms and Health Summit proceedings.

The Mushroom Council convened the Mushrooms and Health Summit in Washington, DC, on 9-10 September 2013. The proceedings are synthesized in this article. Although mushrooms have long been regarded as health-promoting foods, research specific to their role in a healthful diet and in health promotion has advanced in the past decade. The earliest mushroom cultivation was documented in China, which remains among the top global mushroom producers, along with the United States, Italy, The Netherlands, and Poland. Although considered a vegetable in dietary advice, mushrooms are fungi, set apart by vitamin B-12 in very low quantity but in the same form found in meat, ergosterol converted with UV light to vitamin D2, and conjugated linoleic acid. Mushrooms are a rare source of ergothioneine as well as selenium, fiber, and several other vitamins and minerals. Some preclinical and clinical studies suggest impacts of mushrooms on cognition, weight management, oral health, and cancer risk. Preliminary evidence suggests that mushrooms may support healthy immune and inflammatory responses through interaction with the gut microbiota, enhancing development of adaptive immunity, and improved immune cell functionality. In addition to imparting direct nutritional and health benefits, analysis of U.S. food intake survey data reveals that mushrooms are associated with higher dietary quality. Also, early sensory research suggests that mushrooms blended with meats and lower sodium dishes are well liked and may help to reduce intakes of red meat and salt without compromising taste. As research progresses on the specific health effects of mushrooms, there is a need for effective communication efforts to leverage mushrooms to improve overall dietary quality.

The role of culinary-medicinal mushrooms on human welfare with a pyramid model for human health.

Mushrooms are part of fungal biota characterized by wonder. They rise up from lignocellulosic wastes: yet they become so bountiful and nourishing. Mushrooms are environmentally friendly. They biosynthesize their own food from agricultural crop residues, which would otherwise cause health hazards. The extant records show the continued use of some mushrooms, e.g., Lentinus edodes, Ganoderma lucidum, and Cordyceps sinensis are now centuries old. This review presents a pyramid model for mushroom uses (industries), as food, dietary supplements (tonic), and medicine. A regular intake of mushrooms can make us healthier, fitter, and happier, and help us live longer. The sense of purpose and vision for the mushroom industries is also briefly discussed. A variety of mushrooms have been used traditionally in many different cultures for the maintenance of health and in the prevention and treatment of various diseases. A total of 126 medicinal functions are thought to be produced by medicinal mushrooms (MM) and fungi, including antitumor, immunomodulating, antioxidant, radical scavenging, cardiovascular, anti-hypercholesterolemia, antiviral, antibacterial, anti-parasitic, antifungal, detoxification, hepatoprotective, and anti-diabetic effects. Special attention is paid to mushroom polysaccharides. Many, if not all, higher Basidiomycetes mushrooms contain biologically active polysaccharides in fruit bodies, cultured mycelium, and cultured broth. The data on mushroom polysaccharides are summarized for approximately 700 species of higher Hetero- and Homobasidiomycetes. In particular, the most important for modern medicine are polysaccharides with antitumor and immunostimulating properties. Several of the mushroom polysaccharide compounds have proceeded through phase I, II, and III clinical trials and are used extensively and successfully as drugs in Asia to treat various cancers and other diseases. Mushrooms are superior sources of different types of dietary supplements (DSs) (tonics). The advantages of using mushroom-based DSs as a matter of safety (as opposed to herbal preparations) are: (1) The overwhelming majority of mushrooms used for production of DSs are cultivated commercially (and not gathered in the wild). (2) Mushrooms are easily propagated vegetatively and thus keep to one clone. The mycelium can be stored for a long time, and the genetic and biochemical consistency can be checked after a considerable time. (3) The main advantage, in our opinion, is that many mushrooms are capable of growing in the form of mycelial biomass in submerged cultures. In this review, we discuss legal and regulatory issues introducing and controlling DSs from MMs in different countries, including the United States, the European Community, Australia, New Zealand, Japan, and P.R. China, and guidelines of the World Health Organization. One of the targets of the present review is also to draw attention to many critically important unsolved problems in the future development of medicinal mushroom science in the 21st century.

Development of an antigen-capture enzyme-linked immunosorbent assay using monoclonal antibodies for detecting H6 avian influenza viruses.

The H6 subtype of avian influenza virus (AIV) infection occurs frequently in wild and domestic birds. AIV antigen detection is preferred for controlling AIV as birds are infected before they produce antibodies. The purpose of this study was to develop an early diagnostic method for AIV detection. Six monoclonal antibodies (mAbs) developed from a field H6N1 AIV strain were tested for their ability to bind to viruses. The two that showed the greatest binding ability to AIVs were used for antigen detection. An antigen-capture enzyme-linked immunosorbent assay (ELISA) to detect H6 AIVs was developed using these mAbs. One mAb was coated onto an ELISA plate as the capture antibody. The other mAb was used as the detector antibody after labeling with horseradish peroxidase. The antigen-capture ELISA detected H6N1 AIVs but not H5 AIVs, human H1N1, H3N2 influenza or other viruses. This antigen-capture ELISA could be used to specifically detect H6N1 AIV.

Removal of N-terminal methionine from recombinant proteins by engineered E. coli methionine aminopeptidase.

The removal of N-terminal translation initiator Met by methionine aminopeptidase (MetAP) is often crucial for the function and stability of proteins. On the basis of crystal structure and sequence alignment of MetAPs, we have engineered Escherichia coli MetAP by the mutation of three residues, Y168G, M206T, Q233G, in the substrate-binding pocket. Our engineered MetAPs are able to remove the Met from bulky or acidic penultimate residues, such as Met, His, Asp, Asn, Glu, Gln, Leu, Ile, Tyr, and Trp, as well as from small residues. The penultimate residue, the second residue after Met, was further removed if the antepenultimate residue, the third residue after Met, was small. By the coexpression of engineered MetAP in E. coli through the same or a separate vector, we have successfully produced recombinant proteins possessing an innate N terminus, such as onconase, an antitumor ribonuclease from the frog Rana pipiens. The N-terminal pyroglutamate of recombinant onconase is critical for its structural integrity, catalytic activity, and cyto-toxicity. On the basis of N-terminal sequence information in the protein database, 85%-90% of recombinant proteins should be produced in authentic form by our engineered MetAPs.

Medicinal mushroom extracts inhibit ras-induced cell transformation and the inhibitory effect requires the presence of normal cells.

Previously, we developed a simple Rat 6 (R6) cell system by which the inhibitory effects of non-cytotoxic chemicals can be assessed by focus formation assay upon transfection of ras oncogene to the host cells. Using this system, two well studied medicinal mushrooms Ganoderma lucidum and Tricholoma lobayense with anticancer potential were examined for their possible advert effects on cell transformation induced by ras oncogene. Results indicated that both species of mushrooms yielded strong inhibitory effects on ras-induced cell transformation. Further study on T.lobayense indicated that the DEAE-column-bound, polysaccharides (PS)-peptide enriched, but not the unbound fraction, showed strong inhibition in a dosage-dependent manner. Subsequent time course study revealed that the continued presence of the extract in the transfected cultures was required for a maximum inhibitory effect. At the same time, we also observed that significant levels of inhibition occurred even when the application of the extract was delayed until day 12 after transfection. Using a stable transformed cell line, R6/GFP-Ras expressing green fluorescent protein-ras fusion protein in a co-culture assay with normal R6 cells, we demonstrated that R6/GFP-Ras cells grew into green fluorescent foci with striking transforming morphology in the absence of extracts. However, in the presence of extracts, R6/GFP-Ras cells, in most cases, remained as small colonies compiled with only a few green fluorescent cells. Moreover, the inhibitory effect requires the presence of R6 cells. In our study, mushroom extracts have no effect on the growth of individually cultured normal and transformed R6 cells. It is noteworthy that the extracts do not affect the level, or the subcellular localization of the Ras protein. Collectively, the data strongly suggest that the inhibitory effect of the mushroom extracts is not due to a direct killing of the transformed cells, rather, it may be mediated through the surrounding normal R6. While the general understanding of the antitumor effect of PS and PSPC is mediated through the cytokines released by activated macrophages and T-lymphocytes, our data may provide a novel alternative mechanism that the mushroom PS peptides may exert anticancer effect by targeting the ras-mediated signaling pathway.