Formaldehyde as a Chemical Defence Agent of Fruiting Bodies of Mycena rosea and its Role in the Generation of the Alkaloid Mycenarubin†C.

Mycenarubin C, a previously unknown red pyrroloquinoline alkaloid, was isolated from fruiting bodies of the mushroom Mycena rosea and its structure was elucidated mainly by NMR spectroscopy and mass spectrometry. Unlike mycenarubin A, the major pyrroloquinoline alkaloid in fruiting bodies of M. rosea, mycenarubin C, contains an eight-membered ring with an additional C1 unit that is hitherto unprecedented for pyrroloquinoline alkaloids known in nature. Incubation of mycenarubin A with an excess of formaldehyde revealed that mycenarubin C was generated nearly quantitatively from mycenarubin A. An investigation into the formaldehyde content of fresh fruiting bodies of M. rosea showed the presence of considerable amounts of formaldehyde, with values of 5 µg per gram of fresh weight in fresh fruiting bodies. Although mycenarubin C did not show bioactivity against selected bacteria and fungi, formaldehyde inhibits the growth of the mycoparasite Spinellus fusiger at concentrations present in fruiting bodies of M. rosea. Therefore, formaldehyde might play an ecological role in the chemical defence of M. rosea against S. fusiger. In turn, S. fusiger produces gallic acid-presumably to detoxify formaldehyde by reaction of this aldehyde with amino acids and gallic acid to Mannich adducts.

HR-MALDI-MS imaging assisted screening of ß-carboline alkaloids discovered from Mycena metata.

Fruiting bodies of Mycena metata were screened for the presence of new secondary metabolites by means of HPLC-UV, LC-HR-ESIMS, and high-resolution matrix-assisted laser desorption/ionization mass spectrometry imaging (HR-MALDI-MS imaging). Thus, a new ß-carboline alkaloid, 6-hydroxymetatacarboline D (1d), was isolated from fruiting bodies of M. metata. 6-Hydroxymetatacarboline D consists of a highly substituted ß-carboline skeleton, which is likely to be derived biosynthetically from l-tryptophan, 2-oxoglutaric acid, l-threonine, and l-proline. The structure of the alkaloid was established by 2D NMR spectroscopic methods and HR-ESIMS. Moreover, by extensive application of LC-HR-ESIMS, LC-HR-ESIMS/MS, and LC-HR-ESIMS(3) techniques we were able to elucidate the structures of a number of accompanying ß-carboline alkaloids, 1a-1c, 1e-1i, and 2a-2g, structurally closely related to 6-hydroxymetatacarboline D, which are present in M. metata in minor amounts. The absolute configuration of the stereogenic centers of the ß-carboline alkaloids was determined by GC-MS comparison with authentic synthetic samples after hydrolytic cleavage and derivatization of the resulting amino acids.

Mycenaaurin A, an antibacterial polyene pigment from the fruiting bodies of Mycena aurantiomarginata.

A new polyene pigment, mycenaaurin A (1), was isolated from fruiting bodies of Mycena aurantiomarginata. Mycenaaurin A consists of a tridecaketide that is flanked by two amino acid moieties. These are likely to be derived biosynthetically from S-adenosylmethionine. The tridecaketide itself contains an alpha-pyrone, a conjugated hexaene, and an isolated alkenyl moiety. The structure of the new pigment was established by 2D NMR spectroscopic methods and APCIMS. The absolute configuration of the four stereogenic centers was determined by degradation of 1 by ozonolysis and GC-MS comparison of the resulting fragments, after appropriate derivatization, with authentic synthetic samples. Mycenaaurin A (1) might act as a constitutive defense compound, since it exhibits antibacterial activity against the gram-positive bacterium Bacillus pumilus.

Evaluating interventions to improve gait in cerebral palsy: a meta-analysis of spatiotemporal measures.

A number of interventions to improve gait in individuals with cerebral palsy (CP) have been reported in the literature. The aim of this study was to perform a meta-analysis of these studies to determine the overall efficacy of these interventions. Effect sizes (Hedge's g) for spatiotemporal measures of gait (velocity, cadence, stride length) pre- and postintervention were analyzed. Sixty-three studies were included, and the overall effect size was statistically significant for both fixed effects and random models. Types of interventions were grouped into spasticity treatments, orthopedic (bony and soft tissue) surgery, lower extremity orthoses, or 'other'. When the data were analyzed in subgroups by type of intervention, each intervention had a statistically significant effect size with the exception of the 'other'. More importantly, the present study indicates the need to address participant inclusion criteria and power analysis more adequately in future research studies of interventions to improve gait in CP.

Multiple paternity in a salamander with socially monogamous behaviour.

In the majority of birds and mammals, social monogamy is not congruent with genetic monogamy. No research to date has compared social and genetic monogamy in amphibians. We analysed paternity in clutches of red-backed salamanders (Plethodon cinereus), a species in which social monogamy has been demonstrated in the laboratory, and 28% of individuals in the forest are found in male-female pairs in the noncourtship season. We collected 16 clutches of eggs of P. cinereus in the southern Appalachian Mountains of Virginia and collected tail clippings from attending mothers. We genotyped embryos and adults at five microsatellite loci in order to analyse paternity of clutches. Most clutches (84.6%) had multiple sires, with two to three sires per clutch. In this study, 25% of clutches had males in addition to females attending eggs. None of the mothers of these clutches were genetically monogamous. All attending males sired some of the offspring in the clutch that they attended (between 9% and 50%) but never sired a majority in that clutch. We conclude that, at least in this population, social monogamy in P. cinereus is not concomitant with genetic monogamy.

Estimating research productivity and quality in assistive technology: a bibliometric analysis spanning four decades.

OBJECTIVE: Conduct a quantitative assessment of the number of papers contained in MEDLINE related to selected types of assistive technology (AT), and to identify journals publishing significant numbers of papers related to AT, and evaluate them with quantitative productivity and quality measures. DESIGN: Consecutive sample of all papers in MEDLINE identified by standard medical subject headings for selected types of AT from 1963-2003. MAIN OUTCOME MEASURES: Number of journals carrying AT papers, papers per journal (both total number and those specific to AT), journal impact factor, circulation, and number of AT citations per year over time for each area of AT. RESULTS: We present search terms, estimates of the numbers of AT citations in MEDLINE, the journals most likely to contain articles related to AT, journal impact factors, and journal circulations (when available). We also present the number of citations in various areas of AT over time from 1963-2003. Suggestions are presented for possible future modifications of the MEDLINE controlled vocabulary, based on terminology used in existing AT classifications schemes, such as ISO 9999. CONCLUSION: Research papers in the areas of AT examined showed publication across a wide variety of journals. There are a number of journals publishing articles in AT that have impact factors above the median. Some areas of AT have shown an increase in publications per year over time, while others have shown a more constant level of productivity.

Prosthetic device provision to landmine survivors in Bosnia and Herzegovina: outcomes in 3 ethnic groups.

OBJECTIVE: To determine differences in prosthetic provision, use, and effectiveness among unilateral lower-extremity amputees from 3 ethnic groups in Bosnia and Herzegovina. DESIGN: Case series with a consecutive sample of patients seen in field clinics. SETTING: Multiple field clinics in Bosnia and Herzegovina from October 1998 to May 2002. PARTICIPANTS: A total of 671 patients were examined, and information about their prosthetic history was recorded from observation or verbal responses. The majority of the amputations resulted from injuries inflicted by landmines. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Self-reported walking distance per day, prosthesis functionality (functional or nonfunctional as assessed by a physician and a prothetist), prosthesis status (broken or nonbroken as assessed by a physician and a prothetist), and employment status. RESULTS: Ethnic groups differed significantly in types of prostheses provided, functional status of the prostheses, and use of the prostheses for community ambulation. CONCLUSION: Prosthetic devices and delivery of rehabilitation services for unilateral lower-extremity amputees differed between ethnic groups. Despite these differences, functional prosthetic devices increased mobility. Persons in all 3 ethnic groups with functional prostheses were more mobile than persons with nonfunctional prostheses. The employment rate was higher for people with functional prostheses.

Salamanders ( Plethodon cinereus) go for more: rudiments of number in an amphibian.

Techniques traditionally used in developmental research with infants have been widely used with nonhuman primates in the investigation of comparative cognitive abilities. Recently, researchers have shown that human infants and monkeys select the larger of two numerosities in a spontaneous forced-choice discrimination task. Here we adopt the same method to assess in a series of experiments spontaneous choice of the larger of two numerosities in a species of amphibian, red-backed salamanders ( Plethodon cinereus). The findings indicate that salamanders "go for more," just like human babies and monkeys. This rudimentary capacity is a type of numerical discrimination that is spontaneously present in this amphibian.

Breathing induced by abdominal muscle stimulation in individuals without spontaneous ventilation.

The purpose of the study was to determine if functional electrical stimulation of abdominal muscles (FESAM) could maintain pulmonary ventilation at acceptable levels in individuals with spinal cord injury (SCI) who are unable to breathe spontaneously. This is the first published investigation of this technique in this subject population. This case series study included three individuals with SCI; two were on mechanical ventilation (MV), and one used mechanical ventilation and also had an implanted phrenic nerve stimulator (PNS). Using surface electrodes, stimulation was applied to the rectus abdominis and lateral group of abdominal muscles. Repetitive trains of pulses produced a breathing frequency of 20 breaths/min. The longest periods of breathing using only FESAM-supported ventilation for the three subjects were 30, 40, and 210 s, respectively. Airflow at the mouth and volumes were measured with a pneumotachograph and/or optoelectronic plethysmography. Oxygenation was monitored with a pulse oximeter. The tidal volumes generated exclusively by FESAM were sufficient to maintain adequate oxygenation during the periods of stimulation. When oxygenation measured with pulse oximetry dropped to 92% saturation, FESAM was discontinued, and MV or PNS was resumed. This is the first report of achieving successful ventilation in individuals with SCI who have zero tidal volume using FESAM. These preliminary results indicate the clinical potential of FESAM as an additional tool in the armamentarium of supported ventilation.

Diet diversity and clutch size of aquatic and terrestrial salamanders.

Terrestrial species of salamanders generally have a higher diversity of prey in their stomachs and produce smaller and less frequent clutches than do species living in wetter habitats. This may be a consequence of differences in prey availability in the two types of habitats. Prey frequently fluctuate in availability in terrestrial areas as a result of fluctuations in rainfall and the inability of salamanders to forage efficiently during dry periods; the salamanders respond with a generalist diet and relatively K-selected reproductive tactics. In wetter habitats, prey fluctuations are probably dampened due to more constant conditions of moisture; salamanders respond with a more specialized diet and relatively r-selected reproductive tactics.

Fluctuations in prey availability and food limitation for a terrestrial salamander.

A Virginia population of the forest-dwelling salamander Plethodon cinereus was repeatedly sampled over four years to determine volume and number of prey ingested under varying conditions of moisture and temperature. The proportion of the population on a daily negative energy budget was estimated by comparing actual foraging success with laboratory determined values of energetic requirements at various temperatures.Prey became "limited" in availability during rainless periods, apparently because salamanders were not able to forage in dry leaf litter. Foraging success increased with increasing rainfall. Food was a "limiting" resource for a majority of the population on most sampling days, as determined by energy budget analysis. While ambient moisture regulated food availability, ambient temperature set the metabolic requirements and assimilation efficiencies for the population. Consequently, food was more limiting on dry, warm days and less so on wet, cool days. These data support the hypothesis that intraspecific competition frequently occurs for a food resource that is periodically limited in availability.

Competitive Exclusion as a Factor Influencing the Distributions of Two Species of Terrestrial Salamanders.

The salamander Plethodon richmondi shenandoah is restricted to areas of talus on Hawksbill Mountain, Shenandoah National Park, Virginia, while P. cinereus inhabits the soil outside the talus. To test the hypothesis that the former species is excluded from the soil by the latter species, three enclosures were constructed in each of four habitats: (i) in talus consisting only of bare rocks, (ii) in talus where soil has infiltrated between but not under rocks, (iii) in isolated pockets of shallow soil within the rocky confines of the talus, and (iv) in deep soil outside the talus. In each habitat one enclosure contained an isolated sample of shenandoah, another an isolated sample of cinereus, and the third a mixture of the two species. The above testes indicated that: (i) neither species survived the extremely dry conditions of the bard rocks for more than a week, (ii) shenandoah expressed a higher survivorship than did cinereus in this relatively dry habitat, (iii) in the shallow soil shenandoah had lower survivorship in the presence of cinereus than it did in isolation, and (iv) in the deep soil shenandoah had poorer survivorship than cinereus in the mixed species enclosure and poorer survivorship than its isolated control. P. cinereus apparently inhibits the presence of shenandoah in areas of soil, while shenandoah survives better under rocky conditions. However, the talus is a suboptimal habitat for shenandoah, since its survivorship in isolated enclosures is significantly better in the presence of soil. It seems unlikely that salamanders other than cinereus could exclude shenandoah from the soil or that other animals exert a strong influence. Predation, diseases, and parasites also do not appear to exclude shenandoah from areas of deep soil. Competitive exclusion by cinereus has probably restricted the distribution of shenandoah to the suboptimal talus refugium.