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Identification of mushroom body miniature, a zinc-finger protein implicated in brain development of Drosophila.

The mushroom bodies are bilaterally arranged structures in the protocerebrum of Drosophila and most other insect species. Mutants with altered mushroom body structure have been instrumental not only in establishing their role in distinct behavioral functions but also in identifying the molecular pathways that control mushroom body development. The mushroom body miniature(1) (mbm(1)) mutation results in grossly reduced mushroom bodies and odor learning deficits in females. With a survey of genomic rescue constructs, we have pinpointed mbm(1) to a single transcription unit and identified a single nucleotide exchange in the 5' untranslated region of the corresponding transcript resulting in a reduced expression of the protein. The most obvious feature of the Mbm protein is a pair of C(2)HC zinc fingers, implicating a function of the protein in binding nucleic acids. Immunohistochemical analysis shows that expression of the Mbm protein is not restricted to the mushroom bodies. BrdUrd labeling experiments indicate a function of Mbm in neuronal precursor cell proliferation.

5' Untranslated Regions↗

Diagnostic accuracy of urinary amanitin in suspected mushroom poisoning: a pilot study.

BACKGROUND: Amatoxin-containing species are responsible for the most severe cases of mushroom poisoning, with high mortality rate. Therefore, this poisoning should be ruled out in all patients presenting gastrointestinal symptoms after wild mushroom ingestion. OBJECTIVE: To determine sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic efficacy (DE) of urinary amanitin analysis in cases of suspected mushroom poisoning. METHODS: All cases of mushroom ingestion referred to a Poison Center during a one-month period were analyzed. Amanitin measurements were performed by ELISA method (functional least detectable dose 1.5 ng/ml; cut-off value not clearly established). Gastrointestinal symptoms latency and initial clinical assessment were considered alternative diagnostic tools. Definitive diagnosis was used as the reference standard. RESULTS: Among 61 patients included in the study, amatoxin poisoning was diagnosed in 10 cases. Urine samples were collected 5.5 to 92 hours after mushroom ingestion. Urinary amanitin DE was 91.8%, 93.4%, and 80.3%, based on the cut-off value considered (1.5, 5.0, and 10.0 ng/ml, respectively). Symptoms latency longer than 6 hours and initial clinical assessment DE were 70.5% and 67.2%, respectively. To identify amatoxin poisoning, initial clinical assessment resulted more sensitive and urinary amanitin analysis more specific. CONCLUSIONS: Urinary amanitin analysis is a valuable diagnostic tool and may significantly contribute to the management of suspected mushroom poisoning. At present, the best diagnostic accuracy can be obtained taking advantage of both the high sensitivity and negative predictive value of the clinical assessment performed by an experienced toxicologist, and the high specificity and positive predictive value that characterize urinary amanitin analysis.

Adult↗

[Descriptive epidemiology of mushroom poisoning in Japan].

The Incidence of mushroom poisoning was surveyed statistically from 1959 to 1988 in Japan. The results are summarized as follows: 1. During the past three decades, the total number of incidents of mushroom poisoning was 2,096, which involved 10,924 patients and 72 deaths. The average number of incidents was 70 cases per year, involving 364 patients and 2.4 deaths, and the number of incidents decreased gradually every year. Mushroom poisoning usually happened most frequently in September and October. 2. Considering regional differences, the incidence of mushroom poisoning was more frequent in the northeastern part of Japan than in the southwestern part. The incidences of mushroom poisoning in the prefectures of Nagano, Hokkaido, Niigata, Iwate and Fukushima were relatively high. 3. Three species of mushrooms, L. japonicus, R. rhodopolius (R. sinuatus) and T. ustale caused the majority of all poisonings. 4. The rates of total patients and fatalities for each type of poisoning, which were classified according to the symptoms caused, were 90.3% and 10.7% in the type with cholera-like symptoms, 90.2% and 0.2% in that with gastro-intestinal irritation, and 74.1% and 0% in that with neurological symptoms, respectively.

Humans↗

Chronic hypersensitivity pneumonitis induced by Shiitake mushroom spores associated with lung cancer.

A 61-year-old man was admitted to our hospital with a 6-month history of productive cough. He, along with his wife, had been involved with Shiitake mushroom cultures for a period of 12 years. On admission, chest radiography showed bilateral fine-nodular shadow and CT scans showed reticulonodular opacities and a ground-glass appearance predominantly in the subpleural area in both lungs, and a mass in the left S6. Resected pathological specimens obtained by left lower lobectomy revealed lung adenosquamous carcinoma (stage IB), interstitial changes accompanied with lymphocyte proliferation and fibrosis, and granuloma with giant cells. Serum precipitins for Shiitake mushroom antigens were positive. The productive cough improved after the hospital admission and occurred again when he returned to work with the Shiitake mushroom production. Therefore, chronic hypersensitivity pneumonitis (HP) caused by Shiitake mushroom spores was diagnosed. Moreover, his wife was found to have HP caused by mushrooms at this time. There are only two previous reports of chronic HP caused by Shiitake mushroom in Japan, and this is the first case of chronic HP associated with lung cancer.

Alveolitis, Extrinsic Allergic↗

[Differential diagnosis of mushroom poisoning].

Mushroom poisoning is a very urgent problem of internal medicine. Mushroom picking is one of the most popular hobbies in the Czech and Slovak Republic. This is one of the main reasons of the rising incidence of mushroom poisoning, incl. fatal cases. Mushroom poisoning is characterized by a very varied clinical picture which is reflected also in recent classification systems. The main objective of the present work is to draw attention to cardinal symptoms and signs of different types of mushroom poisoning which respect new findings in the diagnosis and differential diagnosis of mushroom poisoning.

Diagnosis, Differential↗

[The forensic chemical diagnosis of mushroom poisonings].

Forensic chemical diagnosis of mushroom poisonings is a problem still to be solved. We propose qualitative tests (color and microcrystal) and thin-layer chromatography for identification of mushroom toxins belonging to alkaloids, aromatic amines, hydrazines, and phenols in fresh mushroom tissue and in biological material. Comparative analysis of the composition of edible and toxic mushrooms revealed the presence of toxins in both groups. Methods for identification of the toxic components and measurements of their concentrations expressed as relevant indexes (micrograms of a toxic component per gram of fresh mushroom tissue) are proposed. A method of forensic chemical differential diagnosis of poisoning with the most prevalent toxic mushrooms is offered.

Basidiomycota↗

Mushroom poisoning in children: report of an accident.

There were very few cases of mushroom poisoning every year in Taiwan and yet no fatal incident have been reported. A species named Amanita phalloides had high lethality rate (22-33%). Physicians should be aware of it's toxic property and be able to treat the patient in case of mushroom poisoning. However, mycologist sometimes might not distinguish edible from toxic mushroom so that educating people not to ingest wild mushroom is very important. We present 4 children with gastrointestinal syndrome after ingesting wild mushroom. We also introduce some basic approaches in treating mushroom poisoning in this paper.

Accidents, Home↗

Larval and pupal development of the mushroom bodies in the honey bee, Apis mellifera.

The mushroom bodies are paired neuropils in the insect brain that act as multimodal sensory integration centers and are involved in learning and memory. Our studies, by using 5-bromo-2-deoxyuridine incorporation and the Feulgen technique, show that immediately before pupation, the brain of the developing honey bee (Apis mellifera) contains approximately 2,000 neuroblasts devoted to the production of the mushroom body intrinsic neurons (Kenyon cells). These neuroblasts are descended from four clusters of 45 or fewer neuroblasts each already present in the newly hatched larva. Subpopulations of Kenyon cells, distinct in cytoarchitecture, position, and immunohistochemical traits, are born at different, but overlapping, periods during the development of the mushroom bodies, with the final complement of these neurons in place by the mid-pupal stage. The mushroom bodies of the adult honey bee have a concentric arrangement of Kenyon cell types, with the outer layers born first and pushed to the periphery by later born neurons that remain nearer the center of proliferation. This concentricity is further reflected in morphologic and immunohistochemical traits of the adult neurons, and is demonstrated clearly by the pattern of expression of Drosophila myocyte enhancer factor 2 (DMEF2)-like immunoreactivity. This is the first comprehensive study of larval and pupal development of the honey bee mushroom bodies. Similarities to patterns of neurogenesis observed in the mushroom bodies of other insects and in the vertebrate cerebral cortex are discussed.

Animals↗

Mushroom bodies of vespid wasps.

Mushroom bodies are higher centers in the brains of insects. Studies on honey bees and species of ants suggest that these centers are particularly prominent in social insects. The present study confirms the presence of large mushroom bodies in five subfamilies of vespid wasps, while at the same time showing significant departures from the mushroom body organization that typifies bees and ants. Although the basic organizational plan of the insect mushroom body into calyces, peduncle, and lobes is maintained, as is the arrangement of axons of intrinsic neurons, the size and arrangements of the vespid mushroom body lobes differ markedly from those known from other Hymenoptera. Furthermore, considerable variation is found both between and within vespid subfamilies. The present results are discussed with respect to current hypotheses about functional attributes of mushroom bodies and the phylogeny of the Vespidae.

Animals↗

Concentrated expression of Ca2+/ calmodulin-dependent protein kinase II and protein kinase C in the mushroom bodies of the brain of the honeybee Apis mellifera L.

We have previously used the differential display method to identify a gene that is expressed preferentially in the mushroom bodies of worker honeybees and to show that it encodes a putative inositol 1,4,5-trisphosphate receptor (IP3R) homologue (Kamikouchi et al. [1998] Biochem. Biophys. Res. Commun. 242:181-186). In the present study, we examined whether the expression of some of the genes for proteins involved in the intracellular Ca2+ signal transduction is also concentrated in the mushroom bodies of the honeybee by isolating cDNA fragments that encode the Ca2+/calmodulin-dependent protein kinase II (CaMKII) and protein kinase C (PKC) homologues of the honeybee. In situ hybridization analysis revealed that the expression of these genes was also concentrated in the mushroom bodies of the honeybee brain: The CaMKII gene was expressed preferentially in the large-type Kenyon cells of the mushroom bodies, whereas that for PKC was expressed in both the large and small types of Kenyon cells. The expression of the genes for IP3R and CaMKII was concentrated in the mushroom bodies of the queen and drone as well as in those of the worker bee. Furthermore, the enzymatic activities of CaMKII and PKC were found to be higher in the mushroom bodies/central bodies than in the optic and antennal lobes of the worker bee brain. These results suggest that the function of the intracellular Ca2+ signal transduction is enhanced in Kenyon cells in comparison to other neuronal cell types in the honeybee brain.

Amino Acid Sequence↗

Subdivision of the Drosophila mushroom bodies by enhancer-trap expression patterns.

Phylogenetically conserved brain centers known as mushroom bodies are implicated in insect associative learning and in several other aspects of insect behavior. Kenyon cells, the intrinsic neurons of mushroom bodies, have been generally considered to be disposed as homogenous arrays. Such a simple picture imposes constraints on interpreting the diverse behavioral and computational properties that mushroom bodies are supposed to perform. Using a P[GAL4] enhancer-trap approach, we have revealed axonal processes corresponding to intrinsic cells of the Drosophila mushroom bodies. Rather than being homogenous, we find the Drosophila mushroom bodies to be compound neuropils in which parallel subcomponents exhibit discrete patterns of gene expression. Different patterns correspond to hitherto unobserved differences in Kenyon cell trajectory and placement. On the basis of this unexpected complexity, we propose a model for mushroom body function in which parallel channels of information flow, perhaps with different computational properties, subserve different behavioral roles.

Animals↗

A distinctive ribonuclease from fresh fruiting bodies of the medicinal mushroom Ganoderma lucidum.

A ribonuclease with an N-terminal sequence distinct from other mushroom ribonucleases was isolated from fresh fruiting bodies of the medicinal mushroom Ganoderma lucidum. The ribonuclease was adsorbed on DEAE-cellulose and Q-Sepharose, and unadsorbed on CM-Sepharose. It possessed a molecular mass of 42 kDa as judged by gel filtration by fast protein liquid chromatography on Superdex 75 and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its molecular mass was similar to that of straw mushroom ribonuclease but much higher compared with those of other mushroom ribonucleases. The ribonuclease was unique among mushroom ribonucleases in that it exhibited the highest potency toward poly(U), followed by poly(A). Its activity toward poly(G) and poly(C) was about one-half of that toward poly(A) and one-quarter of that toward poly(U). A pH of 4.0 and a temperature of 60 degrees C were required for optimal activity of the enzyme. The optimum pH was low compared with those reported for other mushroom ribonucleases.

Agaricales↗

Analysis of agaritine in mushrooms and in agaritine-administered mice using liquid chromatography-tandem mass spectrometry.

A sensitive and specific method for quantifying a genotoxic hydrazine, agaritine, has been developed using liquid chromatography-electrospray ionization tandem mass spectrometry (MS). Synthetic agaritine was structurally assigned by (1)H, (13)C and two-dimensional nuclear magnetic resonance (NMR) analysis (heteronuclear multiple-bond correlation [HMBC] and heteronuclear multiple-quantum coherence [HMQC]), high-resolution fast-atom-bombardment (HR-FAB) MS. Agaritine was separated on an ODS column using 0.01% AcOH-MeOH (99:1) as an eluent with a simple solid-phase-extraction cleanup for mushroom samples and with acetonitrile and methanol deprotenization for plasma samples. There were no interference peaks in any of the mushrooms or mouse plasma samples. The recoveries of agaritine from the spiked mushroom samples and spiked mouse plasma were 60.3-114 and 74.4%, respectively. The intra-day precision values for the spiked mushrooms were 5.5 and 4.2%, and the inter-day precision values were 15.0 and 23.0%, respectively. The limit of quantification was 0.01 microg/g (in mushrooms) and 0.01 microg/ml (in plasma). A precursor ion scan confirmed that agaritine derivatives, which can exert a similar toxicity, were absent. These results indicate that this analytical method for quantifying agaritine could help to evaluate the risk of mushroom hydrazines to humans.

Agaricales↗

Metabolomic discrimination of different grades of pine-mushroom (Tricholoma matsutake Sing.) using 1H NMR spectrometry and multivariate data analysis.

Metabolomic analysis of raw and cooked pine-mushrooms (Tricholoma matsutake Sing.) of different grades was performed using (1)H nuclear magnetic resonance (NMR) spectrometry and principal component analysis (PCA). PCA of the (1)H NMR spectra of aqueous fractions allowed different grades of raw pine-mushroom to be discriminated by a combination of principal component (PC) 1 and PC 2, which accounted cumulatively for 94.1% of the variation in all variables. The major peaks in the (1)H NMR spectra that contributed to discrimination of raw mushrooms were assigned to choline, trehalose, threonine, leucine/isoleucine, succinic acid, alanine, and fumaric acid. The combination of PC 1 (70.8%) and PC 3 (7.5%) allowed different grades of cooked pine-mushroom to be discriminated, and the major peaks in the (1)H NMR spectra that contributed to discrimination of cooked mushrooms were assigned to succinic acid, trehalose, and fumaric acid. This metabolomic analysis-based method allows different grades of pine-mushroom to be distinguished without any prepurification.

Agaricales↗

Dose- and time-dependent hypocholesterolemic effect of oyster mushroom (Pleurotus ostreatus) in rats.

The effect of the dose of oyster mushroom in the diet (1.0, 2.5, and 5.0%) and of the period of application (8, 16, 28, and 52 wk) on cholesterol accumulation in blood and body organs was studied in weanling male Wistar rats fed a diet containing 0.3% cholesterol. Reduction of cholesterol in serum and body organs was found to be dependent on the amount of dietary oyster mushroom administered. A negative correlation between the mushroom dose and cholesterol level was found after 8 and 28 wk of feeding (r=-0.9821 and -0.9803, respectively; P < 0.02 for both cases). The dose of 1% oyster mushroom did not affect cholesterol levels in serum or body organs. A significant reduction of cholesterol levels was observed in serum (31-46%) and liver (25-30%) at a dose of 5% of oyster mushroom for all periods. Reduced cholesterol content in very low-density lipoproteins (VLDL) was also observed at this level. The highest dose of oyster mushroom induced a decrease in conjugated diene levels in erythrocytes and an increase in the levels of reduced glutathione in the liver and stimulated the activities of catalase and glutathione peroxidase in the liver in the final period of the experiment.

Animals↗

Difference in the volatile composition of pine-mushrooms (Tricholoma matsutake Sing.) according to their grades.

The differences in volatile components of pine-mushrooms (Tricholoma matsutake Sing.) according to their grades were observed by applying multivariate statistical methods to GC-MS data sets. A total of 35 and 37 volatile components were identified in raw and cooked pine-mushrooms, respectively. The volatile components in pine-mushrooms were primarily composed of C8 species, such as 3-octanol, 1-octen-3-ol, 1-octanol, (E)-2-octen-1-ol, 3-octanone, 1-octen-3-one, (E)-2-octenal, and octanoic acid. The levels of ethyl octanoate, junipene, and 3-methyl-3-buten-2-one were much higher in raw pine-mushroom of higher grades, whereas the reverse was true for C8 components. On the other hand, furfuryl alcohol, benzyl alcohol, phenylethyl alcohol, dihydro-5-methyl-2(3H)-furanone, 2(5H)-furanone, (E)-2-methyl-2-butenal, furfural, phenylacetaldehyde, benzoic acid methyl ester, camphene, and beta-pinene were the major components of cooked mushrooms. These volatile components formed by various thermal reactions could be mainly responsible for the difference in volatile components of cooked pine-mushrooms according to their grades.

Agaricales↗

Dose rate effect of gamma irradiation on phenolic compounds, polyphenol oxidase, and browning of mushrooms (Agaricus bisporus).

To enhance the shelf life of edible mature mushrooms, Agaricus bisporus, 2 kGy ionizing treatments were applied at two different dose rates: 4.5 kGy/h (I(-)) and 32 kGy/h (I(+)). Both I(+) and I(-) showed a 2 and 4 day shelf-life enhancement compared to the control (C). Before day 9, no significant difference (p>0.05) in L value was detected in irradiated mushrooms. However, after day 9, the highest observed L value (whiteness) was obtained for the mushrooms irradiated in I(-). Analyses of phenolic compounds revealed that mushrooms in I(-) contained more phenols than I(+) and C, the latter containing the lower level of phenols. The fluctuation of the precursors of glutaminyl-4-hydroxyaniline (GHB) was less in I(-) than in I(+). The polyphenol oxidase (PPO) activities of irradiated mushrooms, analyzed via catechol oxidase, dopa oxidase, and tyrosine hydroxylase substrates, were found to be significantly lowered (p = 0.05) compared to C, with a further decrease in I(+). Analyses of the enzymes indicated that PPO activity was lower in I(+), contrasting with its lower phenols concentration. The observation of mushrooms' cellular membranes, by electronic microscopy, revealed a better preserved integrity in I(-) than in I(+). It is thus assumed that the browning effect observed in I(+) was caused by both the decompartmentation of vacuolar phenol and the entry of molecular oxygen into the cell cytoplasm. The synergetic effect of the residual active PPO and the molecular oxygen, in contact with the phenols, allowed an increased oxidation rate and, therefore, a more pronounced browning I(+) than in I(-).

Agaricales↗

Enzyme inactivation analysis for industrial blanching applications: comparison of microwave, conventional, and combination heat treatments on mushroom polyphenoloxidase activity.

Browning reactions in fruits and vegetables are a serious problem for the food industry. In mushrooms, the principal enzyme responsible for the browning reaction is polyphenoloxidase (PPO). Microwaves have recently been introduced as an alternative for the industrial blanching of mushrooms. However, the direct application of microwave energy to entire mushrooms is limited by the important temperature gradients generated within the samples during heating, which can produce internal water vaporization and associated damage to the mushrooms texture. A microwave applicator has been developed, whereby irradiation conditions can be regulated and the heating process monitored. Whole edible mushrooms (Agaricus bisporus) were blanched by conventional, microwave, and combined heating methods to optimize the rate of PPO inactivation. A combined microwave and hot-water bath treatment has achieved complete PPO inactivation in a short time. Both the loss of antioxidant content and the increase of browning were minor in the samples treated with this combined method when compared to the control. This reduction in processing time also decreased mushroom weight loss and shrinkage.

Agaricales↗