Vertically transmitted microbiome protects eggs from fungal infection and egg failure.

BACKGROUND: Beneficial microbes can be vertically transmitted from mother to offspring in many organisms. In oviparous animals, bacterial transfer to eggs may improve egg success by inhibiting fungal attachment and infection from pathogenic microbes in the nest environment. Vertical transfer of these egg-protective bacteria may be facilitated through behavioral mechanisms such as egg-tending, but many species do not provide parental care. Thus, an important mechanism of vertical transfer may be the passage of the egg through the maternal cloaca during oviposition itself. In this study, we examined how oviposition affects eggshell microbial communities, fungal attachment, hatch success, and offspring phenotype in the striped plateau lizard, Sceloporus virgatus, a species with no post-oviposition parental care. RESULTS: Relative to dissected eggs that did not pass through the cloaca, oviposited eggs had more bacteria and fewer fungal hyphae when examined with a scanning electron microscope. Using high throughput Illumina sequencing, we also found a difference in the bacterial communities of eggshells that did and did not pass through the cloaca, and the diversity of eggshell communities tended to correlate with maternal cloacal diversity only for oviposited eggs, and not for dissected eggs, indicating that vertical transmission of microbes is occurring. Further, we found that oviposited eggs had greater hatch success and led to larger offspring than those that were dissected. CONCLUSIONS: Overall, our results indicate that female S. virgatus lizards transfer beneficial microbes from their cloaca onto their eggs during oviposition, and that these microbes reduce fungal colonization and infection of eggs during incubation and increase female fitness. Cloacal transfer of egg-protective bacteria may be common among oviparous species, and may be especially advantageous to species that lack parental care.

Molecular lemmings: strategies to avoid when designing BODIPY ferrocene dyads for dye-sensitized solar cell applications.

BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacenes) dyes possess intense absorption profiles that can be exploited in various light harvesting applications. However, redox stability and optimization of frontier molecular orbital energies in these dyes are critical for their successful incorporation into new solar cell materials. This article describes the synthesis and characterization of a family of ß-substituted BODIPY-ferrocene dyads with push-pull architectures. Designed to stabilize the photo-oxidized BODIPY for dye-sensitized solar cell (DSSC) applications, some deleterious electron transfer behaviours emerged when the ferrocene unit was conjugated to electron deficient BODIPYs. These findings are discussed herein.

Chemical constituents and their antibacterial activity from the tropical endophytic fungus Diaporthe sp. F2934.

AIMS: To isolate, characterize and determine the antibacterial activities of compounds produced by the endophytic fungus Diaporthe sp. F2934, cultivated on malt extract agar. METHODS AND RESULTS: The fungus was cultivated aseptically in Petri dishes containing malt extract agar at 25°C for 15 days. Crude extract was obtained from mycelium using ethyl acetate and sonication, and was fractioned using classic chromatography and HPLC. The structures of phomosines and chromanones were established by NMR experiments including HMQC, HMBC and COSY. Their molecular formulas were determined by ESI-TOFMS. We obtained six compounds: (1) 4H-1-benzopyra-4-one-2,3-dihydro-5-hydroxy-2,8-dimetyl, (2) 4H-1-benzopyran-4-one-2,3-dihydro-5-hydroxy-8-(hydroxylmethyl)-2-methyl, (3) 4H-1-benzopyra-4-one-2,3-dihydro-5-methoxyl-2,8-dimetyl, (4) phomosine A, (5) phomosine D and (6) phomosine C. Isolated compounds 1, 2 and 5 were inactive against 15 micro-organisms, but phomosines A and C were active against diverse Gram-negative and Gram-positive bacteria. CONCLUSIONS: A group of new chromanones and known phomosines have been isolated from the genus Diaporthe (Diaporthe sp. F2934). The results obtained confirm the wide chemical diversity produced by endophytic fungi, specifically the genus Diaporthe. In addition, phomosines A and C may be considered as antimicrobial agents that can be used to guide the development of new antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: Our phylogenetic analysis places Diaporthe sp. F2934 as sister to the Diaporthe cynaroidis clade. Three chromanones were isolated and identified, for the first time, using crude extract obtained from Diaporthe F2934. From this extract phomosines A, C and D were also purified. Regarding Staphylococcus aureus, the inhibition zone diameter (IZD) for phomosine A was 20% higher than the standard drug, vancomycin. When cultivated as described here, Diaporthe sp. F2934 produced new and antimicrobial compounds.

Chemical and bioactive natural products from Microthyriaceae sp., an endophytic fungus from a tropical grass.

UNLABELLED: In screening for natural products with antiparasitic activity, an endophytic fungus, strain F2611, isolated from above-ground tissue of the tropical grass Paspalum conjugatum (Poaceae) in Panama, was chosen for bioactive principle elucidation. Cultivation on malt extract agar (MEA) followed by bioassay-guided chromatographic fractionation of the extract led to the isolation of the new polyketide integrasone B (1) and two known mycotoxins, sterigmatocystin (2) and secosterigmatocystin (3). Sterigmatocystin (2) was found to be the main antiparasitic compound in the fermentation extract of this fungus, possessing potent and selective antiparasitic activity against Trypanosoma cruzi, the cause of Chagas disease, with an IC50 value of 0.13 µmol l(-1) . Compounds 2 and 3 showed high cytotoxicity against Vero cells (IC50 of 0.06 and 0.97 µmol l(-1) , respectively). The new natural product integrasone B (1), which was co-purified from the active fractions, constitutes the second report of a natural product possessing an epoxyquinone with a lactone ring and exhibited no significant biological activity. Strain F2611 represents a previously undescribed taxon within the Microthyriaceae (Dothideomycetes, Ascomycota). SIGNIFICANCE AND IMPACT OF THE STUDY: The present study attributes new antiparasitic and psychoactive biological activities to sterigmatocystin (2), and describes the structure elucidation of the new natural product integrasone B (1), which possesses a rare epoxyquinone with a lactone ring moiety. This is also the first report of sterigmatocystin (2) isolation in a fungal strain from this family, broadening the taxonomic range of sterigmatocystin-producing fungi. The study also presents taxonomic analyses indicating that strain F2611 is strongly supported as a member of the Microthyriaceae (Ascomycota), but is not a member of any previously known or sequenced genus.

Culture-based study of endophytes associated with rubber trees in Peru reveals a new class of Pezizomycotina: Xylonomycetes.

Through a culture-based survey of living sapwood and leaves of rubber trees (Hevea spp.) in remote forests of Peru, we discovered a new major lineage of Ascomycota, equivalent to a class rank. Multilocus phylogenetic analyses reveal that this new lineage originated during the radiation of the 'Leotiomyceta', which resulted not only in the evolution of the Arthoniomycetes, Dothideomycetes, Eurotiomycetes, Geoglossomycetes, Lecanoromycetes, Leotiomycetes, Lichinomycetes, and Sordariomycetes, but also of the majority of hyperdiverse foliar endophytes. Because its origin is nested within this major burst of fungal diversification, we could not recover strong support for its phylogenetic relationship within the 'Leotiomyceta'. Congruent with their long phylogenetic history and distinctive preference for growing in sapwood, this new lineage displays unique morphological, physiological, and ecological traits relative to known endophytes and currently described members of the 'Leotiomyceta'. In marked contrast to many foliar endophytes, the strains we isolated fail to degrade cellulose and lignin in vitro. Discovery of the new class, herein named Xylonomycetes and originally mis-identified by ITSrDNA sequencing alone, highlights the importance of inventorying tropical endophytes from unexplored regions, using multilocus data sets to infer the phylogenetic placement of unknown strains, and the need to sample diverse plant tissues using traditional methods to enhance efforts to discover the evolutionary, taxonomic, and functional diversity of symbiotrophic fungi.

Fungal endophytes: diversity and functional roles.

All plants in natural ecosystems appear to be symbiotic with fungal endophytes. This highly diverse group of fungi can have profound impacts on plant communities through increasing fitness by conferring abiotic and biotic stress tolerance, increasing biomass and decreasing water consumption, or decreasing fitness by altering resource allocation. Despite more than 100 yr of research resulting in thousands of journal articles, the ecological significance of these fungi remains poorly characterized. Historically, two endophytic groups (clavicipitaceous (C) and nonclavicipitaceous (NC)) have been discriminated based on phylogeny and life history traits. Here, we show that NC-endophytes represent three distinct functional groups based on host colonization and transmission, in planta biodiversity and fitness benefits conferred to hosts. Using this framework, we contrast the life histories, interactions with hosts and potential roles in plant ecophysiology of C- and NC-endophytes, and highlight several key questions for future work in endophyte biology.

Percutaneous coronary intervention without on site surgical back-up; two-years registry of a large Dutch community hospital.

AIMS: To assess safety and efficacy of off-site percutaneous coronary intervention (PCI) in The Dutch invasive cardiovascular system. METHODS AND RESULTS: Descriptive single centre registry of elective and emergency PCI. Setting is a Dutch community hospital, 40 km north of Amsterdam, with an adherent population of 400,000 people. A Clinical follow up of Major Adverse Cardiac and Cerebral Events (MACCE) at 30 days post PCI is performed. The total number of participants eligible for PCI was 781 of whom 545 were men and 236 women. During a two-year period 781 PCI's were performed of which 298 were emergency and 483 elective. Acute complications occurred in 2.1% of participants. MACCE-free was 86.9% in the group with AMI and 95.8% in the elective group. CONCLUSIONS: Off-site PCI is feasible and safe in The Netherlands on the condition that specific key factors for success are taken into consideration.

[Nurse practitioner equally as good as a resident in the treatment of stable patients after recent myocardial infarction, but with more patient satisfaction]. / 'Nurse practitioner' even bekwaam als assistent-geneeskundige voor de behandeling van stabiele patiënten na een recent myocardinfarct, maar met meer tevredenheid bij patiënten.

OBJECTIVE: To determine the feasibility and efficacy ofa nurse-led clinic for stable patients recovering from a recent myocardial infarction, as opposed to a resident-led clinic. DESIGN: Randomized study. METHOD: Over a period of 1 year, data on the treatment and complications of 200 consecutive infarction patients were collected. The patients were randomized on transfer from the coronary-care unit to the cardiology ward. Subsequently, these patients were treated by a registered nurse practitioner (n = 97) or by a resident (n = 103), both of whom were under the direct supervision of the attending cardiologist. Degree of satisfaction was scored by the patients on a 0-10 point scale. RESULTS: The patients in both groups were predominantly men (75%) with a mean age of 63 years. Risk factors and cardiac histories were comparable in both groups as were the location of the infarction and the nature of the acute treatment. No significant differences between the groups were found in the main endpoints: mortality (0%), re-infarctions (2%) or length ofstay. However, patients treated by the nurse practitioner expressed a significantly higher score in the satisfaction study. CONCLUSION: The treatment of stable postmyocardial infarction patients in a nurse-practitioner-led clinic post was found to be feasible and effective with a significantly higher level of patient satisfaction.

Validity of conjoint analysis to study clinical decision making in elderly patients with aortic stenosis.

OBJECTIVE: Written case simulations are increasingly being used to investigate clinical decision making. Our study was designed to determine the validity of written case simulations within a conjoint analysis approach. STUDY DESIGN AND SETTING: We developed a series of 32 written case simulations that differed with respect to nine clinical characteristics. These case simulations represented elderly patients with aortic stenosis. The clinical characteristics varied according to a fractional factorial design. We analyzed retrospectively all consecutive patients of 70 years of age or older with an aortic stenosis in three university hospitals. RESULTS: 34 cardiologists from three Dutch hospitals gave their treatment advice to each of these case simulations on a six-point scale (ranging from 'certainly no' to 'certainly yes' to surgical treatment). We compared the influence that the clinical characteristics had on the responses to these case simulations with their influence on the actual treatment decision for 147 actual patients in the same three hospitals. We found a strong agreement. This agreement was only slightly affected by the cut-off value used to dichotomize the treatment advice into a recommendation in favor of or against surgical treatment. CONCLUSION: Written case simulations reflect well how clinicians are influenced by specific clinical characteristics of their patients.

Glossopharyngeal neuralgia with syncope: a case report.

We present the case of a 60-year-old woman with known glossopharyngeal neuralgia who was admitted to hospital because of recurrent syncopes associated with episodes of painful sensations in the caudal region of her tongue. Rhythm observation showed prolonged asystole, which was accompanied by a loss of consciousness. The asystole was preceded by an episode of pain. We concluded that the bradyarrhythmia and syncopes where associated with the glossopharyngeal neuralgia. Because of the life-threatening condition, we inserted a permanent dual-chamber pacing device. After implantation of the pacemaker, the patient had no further syncopes, although she still complained of episodic tongue pain.

The role of predicted intracranial bleeding risk in the choice of reperfusion strategy in patients with acute myocardial infarction.

OBJECTIVES: Intracranial haemorrhage after thrombolytic therapy for acute myocardial infarction occurs in 0.5-3% of patients. Prediction models have been developed to predetermine the intracranial bleeding risk, but have rarely been used for assigning the optimal reperfusion strategy. This might result in the use of thrombolytic therapy when primary PTCA would be preferable. METHODS: Prospective data were gathered in 1365 candidates for reperfusion therapy. Risk of intracranial haemorrhage was determined with a risk score derived from large-scale clinical trials. Patients were divided into three groups based on their risk of intracranial haemorrhage: <1%, 1-3% and >3% and stratified by age. RESULTS: An intracranial bleeding risk exceeding 3% was found in 120 patients (9%). These high-risk patients were often treated with thrombolysis (87%). Intracranial bleeding actually occurred in four out of 120 patients (3.3%) in this highest risk group, while no bleeding occurred in the other risk groups. CONCLUSION: The actual incidence of intracranial bleeding is similar to the predicted bleeding risk in high-risk patients. These high-risk patients are predominantly older than 70 years. Nearly all patients exceeding a 3% risk of intracranial haemorrhage were treated with thrombolytic therapy. Primary angioplasty should be preferred in patients aged over 70 years since success rates of direct PTCA are no worse in elderly compared with younger patients.

Impaired glucose metabolism predicts mortality after a myocardial infarction.

Diabetes is a risk factor for increased mortality after a myocardial infarction. Whether this applies for patients with hyperglycemia during the acute phase of a acute myocardial infarction is unclear. Therefore we determined the relation between admission plasma glucose level and mortality in a prospectively collected series of 336 consecutive AMI patients. Patients were divided in four groups based on WHO criteria for glucose levels: I: <5.6 mmol/l, II: 5.6--8.3 mmol/l, III: 8.4--11.0 mmol/l, IV: 11.1 mmol/l. The average age was 68+/-11 years with a peak CK of 1378+/-160 U/l, 34% were anterior wall AMIs and 52% were treated with thrombolysis. All patients had a long-term follow-up control at an average of 14.2 months. One year mortality rate was 19.3% and rose to 44% in patients with glucose levels >11.1 mmol/l. The mortality was higher in diabetic patients than in non-diabetic patients (40 vs. 16%; P<0.05). Multivariate analysis revealed an independent effect of glucose level on mortality. In conclusions, our study in an unselected patient population demonstrates that admission plasma glucose level independently predicts 1 year mortality even in absence of diagnosed diabetes mellitus. Further studies evaluating the effect of acute insulin intervention in reducing mortality are warranted.

Variability in treatment advice for elderly patients with aortic stenosis: a nationwide survey in The Netherlands.

OBJECTIVE: To determine how the decisions of Dutch cardiologists on surgical treatment for aortic stenosis were influenced by the patient's age, cardiac signs and symptoms, and comorbidity; and to identify groups of cardiologists whose responses to these clinical characteristics were similar. DESIGN: A questionnaire was produced asking cardiologists to indicate on a six point scale whether they would advise cardiac surgery for each of 32 case vignettes describing 10 clinical characteristics. SETTING: Nationwide postal survey among all 530 cardiologists in the Netherlands. RESULTS: 52% of the cardiologists responded. There was wide variability in the cardiologists' advice for the individual case vignettes. Six groups of cardiologists explained 60% of the variance. The age of the patient was most important for 41% of the cardiologists; among these, 50% had a high and 50% a low inclination to advise surgery. A further 24% were influenced equally by the patient's age and by the severity of the aortic stenosis and its effect on left ventricular function; among these, 62% had a high and 38% a low inclination to advise surgery. Finally, 23% of the cardiologists were mainly influenced by the left ventricular function and 12% by the aortic valve area. The presence of comorbidity always played a minor role. CONCLUSIONS: There were systematic differences among groups of cardiologists in their inclination to advise aortic valve replacement for elderly patients, as well as in the way their advice was influenced by the patients' characteristics. These results indicate the need for prospective studies to identify the best treatment for elderly patients according to their clinical profile.

'Expected infarct size without thrombolysis', a concept that predicts immediate and long-term benefit from thrombolysis for evolving myocardial infarction.

BACKGROUND: Thrombolytic therapy should only be used when expected benefits outweigh the risks. In order to obtain a precise estimation of prognosis, with and without thrombolytic therapy, we postulated that mortality reduction by thrombolytic therapy is a function of the area of myocardium at risk for necrosis. We developed a model to estimate the myocardial area at risk for necrosis from clinical parameters readily available upon hospital admission. This model was validated in relation to long-term prognosis and benefits of thrombolytic therapy. METHODS: Enzymatic infarct size with and without thrombolysis was predicted from the haemodynamic state and the electrocardiogram on hospital admission by multivariate regression analysis in 885 patients in the rt-PA placebo and rt-PA/PTCA trial of the European Cooperative Study Group. This multivariate function was used to validate the 'expected infarct size without thrombolytic treatment' in a test population of 533 patients from the Intracoronary Streptokinase trial of the Interuniversity Cardiology Institute of The Netherlands (ICIN) and 1741 patients from the Intravenous Streptokinase in Acute Myocardial Infarction (ISAM) study, both trials with a non-thrombolysed control group. RESULTS: Expected infarct size correlated well with the actual enzymatic infarct size in the non-thrombolysed patients of the latter two series. Limitation of infarct size by thrombolytic therapy was greatest in patients with a large 'expected infarct size' and absent in patients with a small area at risk. Similarly, one year mortality reduction was greatest in patients with a large 'expected infarct size without thrombolysis'; four deaths were prevented per hundred (95% confidence interval 0 to 9) if the area at risk was large, vs one death (95% confidence interval -2 to 3) in patients with a small area at risk. Benefit was most pronounced in patients with a large area at risk who were treated early within 3 h of symptom onset. A score for the determination of 1 year mortality with and without thrombolytic therapy is presented to help the clinician determine who to treat with thrombolytic therapy. CONCLUSION: 'Expected infarct size without thrombolysis' is a useful tool for clinicians to estimate the amount of myocardium at risk of necrosis in individual patients and to decide whether thrombolytic therapy is warranted. It is the only validated parameter of myocardium at risk for necrosis that is readily available for all patients with myocardial infarction and does not need high-tech equipment.

Modulation of plasma fibrinogen levels by ticlopidine in healthy volunteers and patients with stable angina pectoris.

Elevated plasma fibrinogen levels are associated with an increased risk for cardiac events. Ticlopidine is a drug that inhibits the ADP-induced aggregation of blood platelets and it also has been described that ticlopidine can decrease the plasma fibrinogen level in patients with vascular diseases. The mechanism of this decrease has not yet been elucidated and therefore mechanisms that are known to affect fibrinogen levels were studied, viz, the acute phase reaction, total fibrin plus fibrinogen degradation (TDP) levels and the polymorphisms of the fibrinogen beta-gene. The fibrinogen lowering effect of ticlopidine was studied in 26 healthy volunteers, selected on genotype of the Bcl] polymorphism of the fibrinogen beta-gene, and in 26 patients with stable angina pectoris in a double blind, randomized cross-over study. Functional plasma fibrinogen levels were measured with the Clauss assay. Fibrinogen antigen, C-reactive protein (CRP) and TDP levels were measured using an enzyme immuno assay (EIA). In the healthy volunteers the functional fibrinogen levels had decreased by 0.20 g/l (9%, p = 0.005 using the paired Student l-test) after 4 weeks of 250 mg bid ticlopidine administration, whereas fibrinogen antigen, CRP and TDP levels were not significantly changed. In the stable angina pectoris patients the pre-treatment fibrinogen, CRP and TDP levels were significantly higher than in the volunteer group. After four weeks 250 mg bid ticlopidine administration the functional fibrinogen levels had decreased by 0.38 g/l (11%, p < 0.005), whereas the fibrinogen antigen, CRP and TDP levels were not significantly changed. The levels of functional and antigen fibrinogen, CRP and TDP did not change significantly during the placebo period in the volunteers or the patients. Neither in the volunteers nor in the patients was the effect of ticlopidine on the fibrinogen levels associated with the fibrinogen beta-gene polymorphisms. Therefore, the fibrinogen lowering effect of ticlopidine is likely to be a modulation of the functionality of the molecule and unlikely to be modulated by the acute phase reaction, TDP-levels or the fibrinogen beta-gene polymorphisms.