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Disruption of neurotransmission in Drosophila mushroom body blocks retrieval but not acquisition of memory.

Surgical, pharmacological and genetic lesion studies have revealed distinct anatomical sites involved with different forms of learning. Studies of patients with localized brain damage and work in rodent model systems, for example, have shown that the hippocampal formation participates in acquisition of declarative tasks but is not the site of their long-term storage. Such lesions are usually irreversible, however, which has limited their use for dissecting the temporal processes of acquisition, storage and retrieval of memories. Studies in bees and flies have similarly revealed a distinct anatomical region of the insect brain, the mushroom body, that is involved specifically in olfactory associative learning. We have used a temperature-sensitive dynamin transgene, which disrupts synaptic transmission reversibly and on the time-scale of minutes, to investigate the temporal requirements for ongoing neural activity during memory formation. Here we show that synaptic transmission from mushroom body neurons is required during memory retrieval but not during acquisition or storage. We propose that the hebbian processes underlying olfactory associative learning reside in mushroom body dendrites or upstream of the mushroom body and that the resulting alterations in synaptic strength modulate mushroom body output during memory retrieval.

Animals↗

Effect of packaging conditions on the growth of micro-organisms and the quality characteristics of fresh mushrooms (Agaricus bisporus) stored at inadequate temperatures.

Mushrooms were packed in two polymeric films (perforated and non-perforated PVC) and stored at 17 degrees C and 25 degrees C. The carbon dioxide and oxygen content inside the packages, aerobic mesophiles, Pseudomonas spp., faecal coliforms, Escherichia coli, anaerobic spores and major sensory factors (colour, texture, development stage and presence of moulds) were determined. The non-perforated packages had the highest contents of CO2 (6-7%), the lowest contents of O2 (0.013-0.17%) and the most desirable quality parameters (texture, development stage and absence of moulds). Pseudomonas spp. counts were around 1 logarithmic unit lower in mushrooms packaged in non-perforated film as the O2 concentrations were lower than in perforated film. The mushrooms themselves were inoculated with an enterotoxin A-producing strain of Staphylococcus aureus, packaged in overwrapped trays and stored at 17 and 25 degrees C. Staphylococcus aureus did not grow in the samples stored at 17 degrees C. Only slight growth was observed in mushrooms packaged with non-perforated film after 1 day at 25 degrees C. No enterotoxin was detected in any package. Faecal coliform counts were <2 log cfu g(-1). Escherichia coli was not isolated in any of the samples. At 25 degrees C, counts of anaerobic spores of around 2 log cfu g(-1) were detected in those mushrooms packaged in non-perforated film.

Agaricus↗

Behaviour of Listeria monocytogenes in packaged fresh mushrooms (Agaricus bisporus).

AIMS: The aim of this study was to evaluate the potential of Listeria monocytogenes to grow in mushrooms packaged in two different types of PVC films when stored at 4 degrees C and 10 degrees C. METHODS AND RESULTS: Mushrooms were packed in two polymeric films (perforated and nonperforated PVC) and stored at 4 degrees C and 10 degrees C. The carbon dioxide and oxygen content inside the packages, aerobic mesophiles, psychrotrophs, Pseudomonas spp., Listeria monocytogenes, faecal coliforms, Escherichia coli, anaerobic spores and major sensory factors were determined. The mushrooms packaged in nonperforated film and stored at 4 degrees C had the most desirable quality parameters (texture, development stage and absence of moulds). Listeria monocytogenes was able to grow at 4 degrees C and 10 degrees C in inoculated mushrooms packaged in perforated and nonperforated films between 1 and 2 log units during the first 48 h. After 10 d of storage, the populations of L. monocytogenes were higher in mushrooms packaged in nonperforated film and stored at 10 degrees C. CONCLUSIONS: MAP followed by storage at 4 degrees C or 10 degrees C extends the shelf life by maintaining an acceptable appearance, but allows the growth and survival of L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: According to this study additional hurdles must be studied in order to prevent the growth of L. monocytogenes.

Agaricus↗

Dietary mushroom (Pleurotus ostreatus) ameliorates atherogenic lipid in hypercholesterolaemic rats.

1. The effects of edible oyster mushroom Pleurotus ostreatus on plasma and liver lipid profiles and on the plasma total anti-oxidant status were estimated in hyper- and normocholesterolaemic Long Evans rats. 2. The feeding of 5% powder of the fruiting bodies of P. ostreatus mushrooms to hypercholesterolaemic rats reduced their plasma total cholesterol by approximately 28%, low-density lipoprotein-cholesterol by approximately 55%, triglyceride by approximately 34%, non-esterified fatty acid by approximately 30% and total liver cholesterol levels by > 34%, with a concurrent increase in plasma high-density lipoprotein-cholesterol concentration of > 21%. However, these effects were not observed in mushroom-fed normocholesterolaemic rats. 3. Mushroom feeding significantly increased plasma fatty acid unsaturation in both normo- and hypercholesterolaemic rats. 4. Plasma total anti-oxidant status, as estimated by the oxidation of 2,2'-azino-bis-[3-ethylbenz-thiazoline-6-sulphonic-acid], was significantly decreased in mushroom-fed hypercholesterolaemic rats, concomitant with a decrease in plasma total cholesterol. 5. The present study suggests that 5% P. ostreatus supplementation provides health benefits, at least partially, by acting on the atherogenic lipid profile in the hypercholesterolaemic condition.

Animals↗

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.

Animals↗

Staphylococcal food poisoning caused by imported canned mushrooms.

From February through April 1989, four outbreaks of staphylococcal food poisoning in the United States were associated with eating mushrooms canned in the People's Republic of China (PRC). In the four outbreaks, 99 persons who ate at a suspect facility developed gastrointestinal symptoms within 24 h, including 18 who were hospitalized. Illness was associated with eating mushrooms at a university cafeteria (relative risk [RR] = 53.0), a hospital cafeteria (RR = 13.8), a pizzeria (odds ratio [OR] = infinity), and a restaurant (OR = infinity) (all P < .0001). Staphylococcal enterotoxin A was found by ELISA in mushrooms at the sites of two outbreaks and in unopened cans from the three plants thought to have produced mushrooms implicated in outbreaks. These investigations led to multistate recalls and a US Food and Drug Administration order to restrict entry into the United States of all mushrooms produced in the PRC; until this action, the United States imported approximately 50 million pounds yearly.

Adult↗

Dose due to mushroom ingestion in Spain.

The dose due to ingestion of mushrooms depends on their radioactive content. However, not all the mass of the mushroom ingested is digested and assimilated completely by man. To analyse the degree of assimilation of the radioactive content of mushrooms, we have carried out culture under controlled laboratory conditions for Pleurotus eryngii mushrooms to which we added known activities of 134Cs, 85Sr and 239Pu. We then applied the Van Soest method to the fruiting bodies, which were obtained in order to determine the fraction of these radionuclides associated with the major components of the cell that are assimilable by man. Finally, on the basis of the results, we determined the dose due to 137Cs, 90Sr and (239+240)Pu for several mushroom species collected in different natural and semi-natural ecosystems in Spain.

Agaricales↗

Respiratory disease in a mushroom worker.

A patient with allergic alveolitis demonstrated by lung biopsy is described in a mushroom worker in Washington State presenting with severe airways obstruction. This is the first case of mushroom worker's lung reported in the United States outside the mushroom industry in Pennsylvania. The processes of commercial mushroom growing are discussed, and the literature on mushroom worker's lung is reviewed.

Adult↗

Mushroom plant workers experience a shift towards a T helper type 2 dominant state: contribution of innate immunity to spore antigen.

Contemporary mushroom factories are places where there is a substantial risk of the occurrence of respiratory allergy. The aims of this investigation were to estimate its causative agents and to evaluate the contribution of innate immune response in mushroom workers who cultivate Hypsizigus marmoreus (Bunashimeji). Cross-sectional and follow-up studies were performed in the factory. We investigated CD1b, CD3, CD4, CD8, CD14, CD45RO, CD62L and CD161 expression in peripheral blood mononuclear cells (PBMC) by flow cytometry, and serum levels of interleukin (IL-2), IL-4, granulocyte-macrophage colony stimulating factor (GM-CSF), IL-13 and interferon (IFN)-gamma by enzyme-linked immunosorbent assay (ELISA). Co-culture experiments of PBMC with spore extracts were also performed. Percentages of CD1b+ monocytes, natural killer (NK), NK T and CD4+ T cells were increased in the workers compared with controls. Increases in Th2 type cells, Th2/Th1 ratio and serum IL-13 and decreased IFN-gamma were detected, indicating a Th2-biased status of the workers. The follow-up study showed that monocytes and NK cells increased soon after employment while CD4+ T, Th2 and NK T cells increased gradually as employment time lengthened. Serum precipitating antibody to the mushroom antigen could be detected at a later stage. Co-cultivation of PBMC with the spore extracts induced much higher CD1b expression, and suppressed secretion of Th1 cytokine in culture supernatants. These results indicate that the mushroom antigen contains highly immunogenic substances which stimulate PBMC into a Th2-biased in vivo status, and innate immune cells might also play a critical role in developing respiratory allergy in mushroom workers.

Adult↗

Growth-promoting effect of thermophilic fungi on the mycelium of the edible mushroom Agaricus bisporus.

The growth-promoting effect of the thermophilic fungus Scytalidium thermophilum in mushroom compost on the mycelium of the edible mushroom Agaricus bisporus was investigated. Results obtained by others were confirmed by showing that S. thermophilum leads to an increased hyphal extension rate of the mushroom mycelium. However, it was demonstrated that hyphal extension rates were not clearly related to mushroom biomass increase rates. A number of experiments pointed strongly towards CO2 as the determinant of hyphal extension rates. In compost, CO2 is produced mainly by thermophilic fungi. Several experiments did not reveal any other specific compound produced by S. thermophilum that increases the hyphal extension rate of the mushroom mycelium.

Basidiomycota↗

Characteristics of a hydrated, alginate-based delivery system for cultivation of the button mushroom.

The production of the button mushroom Agaricus bisporus with mycelium-colonized alginate pellets as an inoculant of the growing medium was investigated. Pellets having an irregular surface and porous internal structure were prepared by complexing a mixture of 1% sodium alginate, 2 to 6% vermiculite, 2% hygramer, and various concentrations of Nutrisoy (soy protein) with calcium chloride. The porous structure allowed the pellets to be formed septically and then inoculated and colonized with the fungus following sterilization. By using an enzyme-linked immunosorbent assay (ELISA) to estimate fungal biomass, the matrix components of the pellet were found to be of no nutritive value to A. bisporus. Pellets amended with Nutrisoy at a concentration of 0.5 to 8% supported extensive mycelial growth, as determined by significantly increased ELISA values, with a concentration of 4% being optimal and higher concentrations proving inhibitory. The addition of hydrated, mycelium-invaded pellets to the compost or casing layer supported the thorough colonization of the growing substrate and culminated in the formation of mushrooms that showed normal development and typical morphology. Yields and sizes of mushrooms were comparable from composts seeded with either colonized pellets or cereal grain spawn. Similarly, amending the casing layer with pelletized-mycelium-colonized compost resulted in a 2- to 3-day-earlier and more-synchronous emergence of mushrooms than with untreated casing. This technology shows the greatest potential as a pathogen-free inoculant of the casing layer in the commercial cultivation of mushrooms.

Journal Article↗

Identification of exopolysaccharides produced by fluorescent pseudomonads associated with commercial mushroom (Agaricus bisporus) production.

The acidic exopolysaccharides (EPSs) from 63 strains of mushroom production-associated fluorescent pseudomonads which were mucoid on Pseudomonas agar F medium (PAF) were isolated, partially purified, and characterized. The strains were originally isolated from discolored lesion which developed postharvest on mushroom (Agaricus bisporus) caps or from commercial lots of mushroom casing medium. An acidic galactoglucan, previously named marginalan, was produced by mucoid strains of the saprophyte Pseudomonas putida and the majority of mucoid strains of saprophytic P. fluorescens (biovars III and V) isolated from casing medium. One biovar II strain (J1) of P. fluorescens produced alginate, a copolymer of mannuronic and guluronic acids, and one strain (H13) produced an apparently unique EPS containing neutral and amino sugars. Of 10 strains of the pathogen "P. gingeri," the causal agent of mushroom ginger blotch, 8 gave mucoid growth on PAF. The "P. gingeri" EPS also was unique in containing both neutral sugar and glucuronic acid. Mucoid, weakly virulent strains of "P. reactans" produced either alginate or marginalan. All 10 strains of the pathogen P. tolaasii, the causal agent of brown blotch of mushrooms were nonnmucoid on PAF. Production of EPS by these 10 strains plus the 2 nonmucoid strains of "P. gingeri" also was negative on several additional solid media as well as in two broth media tested. The results support our previous studies indicating that fluorescent pseudomonads are a rich source of novel EPSs.

Alginates↗

Sparsening and temporal sharpening of olfactory representations in the honeybee mushroom bodies.

We explored the transformations accompanying the transmission of odor information from the first-order processing area, the antennal lobe, to the mushroom body, a higher-order integration center in the insect brain. Using Ca2+ imaging, we recorded activity in the dendrites of the projection neurons that connect the antennal lobe with the mushroom body. Next, we recorded the presynaptic terminals of these projection neurons. Finally, we characterized their postsynaptic partners, the intrinsic neurons of the mushroom body, the clawed Kenyon cells. We found fundamental differences in odor coding between the antennal lobe and the mushroom body. Odors evoked combinatorial activity patterns at all three processing stages, but the spatial patterns became progressively sparser along this path. Projection neuron dendrites and boutons showed similar response profiles, but the boutons were more narrowly tuned to odors. The transmission from projection neuron boutons to Kenyon cells was accompanied by a further sparsening of the population code. Activated Kenyon cells were highly odor specific. Furthermore, the onset of Kenyon cell responses to projection neurons occurred within the first 200 ms and complex temporal patterns were transformed into brief phasic responses. Thus two types of transformations occurred within the MB: sparsening of a combinatorial code, mediated by pre- and postsynaptic processing within the mushroom body microcircuits, and temporal sharpening of postsynaptic Kenyon cell responses, probably involving a broader loop of inhibitory recurrent neurons.

Action Potentials↗

Modality-specific segregation of input to ant mushroom bodies.

The mushroom bodies are central brain neuropils involved in the control of complex behavior. In ants, the mushroom bodies are relatively large compared to those of honey bees, whereas the optic lobes of ants are considerably smaller. The general morphology of ant mushroom bodies is similar to that of honey bees. As in other Hymenoptera, the main input region of the mushroom bodies, the calyx, is subdivided into three compartments: the lip, the collar, and the basal ring. In many ant species this compartmentalization is not obvious and can only be visualized using neuronal tracers. The lip region receives antennal input and is large in all ant species. It appears to be composed of at least two different regions that have not yet been characterized in detail. The collar is large in other Hymenoptera, yet in ant workers it varies in size and is always much smaller than the lip region. The collar receives visual input and is relatively larger in males, which generally are more dependant on vision than are workers. The basal ring receives input from both the optic and antennal lobes. In one ant tribe, the Ponerini, the collar region appears to have changed its position, but based on afferent input it appears to be homologous to the hymenopteran collar. Generally, the composition of the mushroom body calyx correlates with the living conditions of ants, reflecting the great importance of olfaction and the lesser and more variable significance of vision for workers of the observed ant species.

Animals↗

Development of the Drosophila mushroom bodies: sequential generation of three distinct types of neurons from a neuroblast.

The mushroom bodies (MBs) are prominent structures in the Drosophila brain that are essential for olfactory learning and memory. Characterization of the development and projection patterns of individual MB neurons will be important for elucidating their functions. Using mosaic analysis with a repressible cell marker (Lee, T. and Luo, L. (1999) Neuron 22, 451-461), we have positively marked the axons and dendrites of multicellular and single-cell mushroom body clones at specific developmental stages. Systematic clonal analysis demonstrates that a single mushroom body neuroblast sequentially generates at least three types of morphologically distinct neurons. Neurons projecting into the (gamma) lobe of the adult MB are born first, prior to the mid-3rd instar larval stage. Neurons projecting into the alpha' and beta' lobes are born between the mid-3rd instar larval stage and puparium formation. Finally, neurons projecting into the alpha and beta lobes are born after puparium formation. Visualization of individual MB neurons has also revealed how different neurons acquire their characteristic axon projections. During the larval stage, axons of all MB neurons bifurcate into both the dorsal and medial lobes. Shortly after puparium formation, larval MB neurons are selectively pruned according to birthdays. Degeneration of axon branches makes early-born gamma neurons retain only their main processes in the peduncle, which then project into the adult gamma lobe without bifurcation. In contrast, the basic axon projections of the later-born (alpha'/beta') larval neurons are preserved during metamorphosis. This study illustrates the cellular organization of mushroom bodies and the development of different MB neurons at the single cell level. It allows for future studies on the molecular mechanisms of mushroom body development.

Animals↗

Hypersensitivity pneumonitis due to mushroom (Pholiota nameko) spores.

Hypersensitivity pneumonitis due to the inhalation of spores from the edible mushroom Pholiota nameko was demonstrated. A 52-year-old mushroom worker noticed cough, exertional dyspnea, and high fever several hours after work in the mushroom-cultivating room. His examination revealed bilateral basilar crepitations, fine granular shadows in both lung fields of chest x-ray film, and slight decrease in diffusion capacity. Precipitin and macrophage migration inhibition factor were found against the mushroom spore extract and not other contaminants and molds. Histologic finding by transbronchial lung biopsy showed granulomatous alveolitis. Manifest increase of lymphocytes was found in his bronchoalveolar lavage fluid. Provocative inhalation test with the extract revealed similar symptoms and signs in his workplace eight hours after inhalation. It seems to be the first case of hypersensitivity pneumonitis to the Pholiota nameko mushroom spores.

Agriculture↗

Insulin-releasing and insulin-like activity of Agaricus campestris (mushroom).

Agaricus campestris (mushroom) has been documented as a traditional treatment for diabetes. Here the administration of mushroom in the diet (62.5 g/kg) and drinking water (2.5 g/l) countered the hyperglycaemia of streptozotocin-diabetic mice. An aqueous extract of mushroom (1 mg/ml) stimulated 2-deoxyglucose transport (2.0-fold), glucose oxidation (1.5-fold) and incorporation of glucose into glycogen (1.8-fold) in mouse abdominal muscle. In acute 20 min tests, 0.25-1 mg/ml aqueous extract of mushroom evoked a stepwise 3.5- to 4.6-fold stimulation of insulin secretion from the BRIN-BD11 pancreatic B-cell line. This effect was abolished by 0.5 mM diazoxide and prior exposure to extract did not affect subsequent stimulation of insulin secretion by 10 mM L-alanine, thereby negating a detrimental effect on cell viability. The effect of extract was potentiated by 16.7 mM glucose, L-alanine (10 mM) and IBMX (1 mM), and a depolarising concentration of KCl (25 mM) did not augment the insulin-releasing activity of mushroom. Activity of the extract was found to be heat stable, acetone soluble and unaltered by exposure to alkali, but decreased with exposure to acid. Dialysis to remove components with molecular mass < 2000 Da caused a 40% reduction in activity. Sequential extraction with solvents revealed insulin-releasing activity to be greatest in polar fractions. Lack of haemagglutinin activity with extract activity indicated that activity was unlikely to be due to a lectin-mediated event. These results demonstrate the presence of antihyperglycaemic, insulin-releasing and insulin-like activity in A. campestris.

1-Methyl-3-isobutylxanthine↗

Comparison of balloon- and mushroom-retained large-bore gastrostomy catheters.

OBJECTIVE: We conducted a retrospective study to evaluate two different types of percutaneous fluoroscopic gastrostomy procedures and catheters. MATERIALS AND METHODS: Between July 8, 1999, and August 4, 2000, 80 percutaneous gastrostomy catheters were placed in 80 patients in 80 attempts. Twenty-five 16-, 18-, or 20-French balloon-retained catheters and 55 20-French mushroom-retained catheters were inserted. Typically, the type of catheters placed was based on operator preference. However, balloon-retained tubes were preferred for use in patients with obstructive head and neck or esophageal malignancies, and mushroom-retained catheters were preferred for use in demented or combative patients. Follow-up was conducted through chart reviews and telephone interviews. The technical success, procedural complications, and catheter complications were recorded. Chi-square statistical analysis was performed. RESULTS: Technical success was 100% (80/80 patients), and no procedural complications occurred. In patients who received balloon-retained catheters, the major complication rate was 0%, the minor complication rate was 8% (2/25 patients), and the tube complication rate was 68% (17/25 patients). The following complications occurred: catheter dislodgment (n = 17), superficial cellulitis (n = 1), and bleeding gastric ulcer (n = 1). In patients who received mushroom-retained catheters, the major complication rate was 0%, the minor complication rate was 3.6% (2/55 patients), and the tube complication rate was 3.6% (2/55 patients). The following complications occurred: superficial cellulitis (n = 2), tube occlusion (n = 1), and peristomal tube leakage (n = 1). No significant differences in major or minor complications were found between the gastrostomy procedures. Balloon-retained catheters had a significantly higher rate of tube complications (p < 0.001). CONCLUSION: Compared with balloon-retained catheters, mushroom-retained gastrostomy catheters are significantly more durable, more secure, and less prone to tube dysfunction. Mushroom-retained catheters should be the preferred type of gastrostomy catheter to place in patients whenever possible.

Catheterization↗