Influence of chicory inulin on the survival of microbiota of a probiotic fermented milk during refrigerated storage

Inulin-supplemented and control probiotic fermented milks were produced using a dairy starter culture containingsStreptococcus thermophilus CHCC 742/2130,Lactobacillus acidophilus La-5 andBifidobacterium lactis Bb-12, and were then stored at 4°C for 6 weeks. Microbiological analyses were performed at weekly intervals. The results showed that the presence of inulin at 1 t o 5% (wt/v) did not influence significantly (P>0.05) the survival rates of eitherS. thermophilus orL. acidophilus. However, the addition of inulin at 5% (wt/v) had a significant beneficial effect (P<0.05) on the viability of bifidobacteria after 28 days of refrigerated storage.     

Microtubule assembly-derived by dimerization of TPPP/p25. Evaluation of thermodynamic parameters for multiple equilibrium system from ITC data

BackgroundThe disordered Tubulin Polymerization Promoting Protein/p25 (TPPP/p25) modulates the dynamics and stability of the microtubule system. In this paper the role of dimerization in its microtubule-related functions is established, and an approach is proposed to evaluate thermodynamic constants for multiple equilibrium systems from ITC measurements.MethodsFor structural studies size exclusion chromatography, SDS-PAGE, chemical cross-linking, circular dichroism, fluorescence spectroscopy and isothermal titration calorimetry were used; the functional effect was analyzed by tubulin polymerization assay. Numerical simulation of the multiple equilibrium was performed with Mathematica software.ResultsThe dimerization of TPPP/p25 is promoted by elevation of the protein concentration and by GTP addition. The dimeric form displaying enhanced tubulin polymerization promoting activity is stabilized by disulfide bond or chemical cross-linking. The GTP binding to the dimeric form (K_d-GTP = 200 μM) is tighter with one order of magnitude than to the monomeric one leading to the enrichment of the dimers. A mathematical model elaborated for the multiple equilibrium of the TPPP/p25-GTP system was validated by fitting the GTP-dependent changes of ellipticity and fluorescence signal in the course of TPPP/p25 titrations. The evaluation of the equilibrium constants rendered it possible to determine the thermodynamic parameters of the association of different TPPP/p25 forms with GTP from ITC measurements.Conclusions/General SignificanceThe dimerization of TPPP/p25 with favorable physiological functional potency is proposed to play significant role in the fine tuning of TPPP/p25-mediated microtubule assembly; the unfolded monomers might be involved in the formation of pathological inclusions characteristic for Parkinson's disease and other synucleinopathies.     

Lifelong foraging and individual specialisation are influenced by temporal changes of resource availability

Resource availability largely determines the distribution and behaviour of organisms. In plant–pollinator communities, availability of floral resources may change so rapidly that pollinator individuals can benefit from switching between multiple resources, i.e. different flowering plant species. Insect pollinator individuals of a given generation often occur in different time windows during the reproductive season. This temporal variation in individual occurrences, together with the rapidly changing resource availability, may lead individuals of the same population to encounter and use different resources, resulting in an apparent individual specialisation. We hypothesized, that 1) individual pollinators change their resource use (flower visitation) during their lifetime according to the changing availability of floral resources, and that 2) temporal variation in individual occurrences of pollinators and in resource availability will partly explain individual specialisation. To test these hypotheses, we observed flower visitations of individually marked clouded Apollo butterflies Parnassius mnemosyne during one reproductive season. We found temporal changes in lifetime individual resource use that followed the changes in resource availability, indicating that butterflies can adjust foraging to varying resource availability. Individuals differed considerably in their resource use. This variation was partly explained by temporal variation in both floral resource availability and temporal occurrence of individual butterflies. We suggest the butterfly as a sequential specialist, i.e. short‐term specialist and long‐term generalist. This foraging plasticity can be essential for short‐living insect pollinators in rapidly changing environments. Although flowering dynamics do not fully explain the variability in foraging, our results highlight the importance of temporal dimension in resource use studies. Ultimately, the relative pace of environmental change compared to individual lifespan may be a key factor in resource use plasticity.     

Societal preferences for the return of incidental findings from clinical genomic sequencing: a discrete-choice experiment

Background:An important challenge with the application of next-generation sequencing technology is the possibility of uncovering incidental genomic findings. A paucity of evidence on personal utility for incidental findings has hindered clinical guidelines. Our objective was to estimate personal utility for complex information derived from incidental genomic findings.Methods:We used a discrete-choice experiment to evaluate participants’ personal utility for the following attributes: disease penetrance, disease treatability, disease severity, carrier status and cost. Study participants were drawn from the Canadian public. We analyzed the data with a mixed logit model.Results:In total, 1200 participants completed our questionnaire (available in English and French). Participants valued receiving information about high-penetrance disorders but expressed disutility for receiving information on low-penetrance disorders. The average willingness to pay was $445 (95% confidence interval [CI] $322–$567) to receive incidental findings in a scenario where clinicians returned information about high-penetrance, medically treatable disorders, but only 66% of participants (95% CI 63%–71%) indicated that they would choose to receive information in that scenario. On average, participants placed an important value ($725, 95% CI $600–$850) on having a choice about what type of findings they would receive, including receipt of information about high-penetrance, treatable disorders or receipt of information about high-penetrance disorders with or without available treatment. The predicted uptake of that scenario was 76% (95% CI 72%–79%).Interpretation:Most participants valued receiving incidental findings, but personal utility depended on the type of finding, and not all participants wanted to receive incidental results, regardless of the potential health implications. These results indicate that to maximize benefit, participant-level preferences should inform the decision about whether to return incidental findings.Clinical genomic sequencing technologies are on the verge of allowing individualized care at reasonable cost.¹ Patients and their families will soon receive information from clinical sequencing that has implications for clinical care, including information on consequences related to disease prognosis, treatment response or hereditary risk for disease.² Clinical sequencing can also generate incidental findings, which are clinically relevant genetic variants for disorders unrelated to the reason for ordering the genetic testing. The decision of whether to provide information about incidental findings is complex because such results will have varying clinical validity (whether the genetic variant causes the disorder) and utility (whether effective medical treatment is available for the disorder).^3,4 For example, although effective medical treatment may be available for some validated incidental findings, other incidental findings may not be validated as causing the disorder, and still others will be validated but not associated with effective treatment options.To address in part the challenges surrounding the return of incidental findings, the American College of Medical Genetics and Genomics published recommendations for reporting incidental findings from clinical sequencing.⁵ The statement lists a minimum of 56 genes that laboratories should examine, with results reported to patients through the managing physician. This list includes genes with high-penetrance mutations (i.e., a high proportion of individuals with the mutation will exhibit clinical symptoms) validated to be associated with disorders for which medical interventions are available.The original version of this statement did not “favour offering the patient a preference” for which results would be returned. The reasoning was that clinicians have a duty to prevent potential harm by telling patients about incidental findings. The working group that developed the recommendations further stated that it is impractical to provide the level of genetic counselling required for informed preference on all potential disorders.⁵ As such, the working group recommended that clinicians discuss with patients the possibility of receiving incidental findings from the list. It was argued that patient autonomy is preserved because patients can decline clinical sequencing if they prefer to not receive information about incidental findings.⁵ However, this rationale has been subject to debate because of its “all-or-none” nature, whereby patients must agree to receive information about incidental findings or clinical sequencing is not provided.^6–9 In April 2014, in response to the ongoing debate, the statement was amended to include an “opt-out” option for patients who do not want to receive information about incidental findings.¹⁰Notwithstanding the ethical debate, there is a lack of quantitative, preference-based economic evidence for the return of incidental genomic findings.⁸ It has been argued⁸ that this gap in evidence hindered development of the working group’s recommendation statement. More generally, evidence on preferences for the return of incidental findings is crucial for health policy, for health systems planning and for informing future lists that may include “many more genes.”⁸ We aimed to generate evidence on the personal utility that study participants from the Canadian public ascribe to the return of incidental genomic findings in the clinical setting. We chose participants from the general public because the public is the largest stakeholder in Canada’s publicly funded health care system.     

Molecular Identification and Antifungal Susceptibilities of Black Aspergillus Isolates from Otomycosis Cases in Hungary

Otomycosis, also known as fungal otitis externa, has been used to describe a fungal infection of the external auditory canal, but sometimes involving the middle ear. Many fungal species have been identified as infectious agents in otomycosis, with Aspergillus and Candida species being the most common. Among aspergilli, Aspergillus niger is the most commonly described species in the literature. In this study, 14 black Aspergillus strains were analyzed, which were isolated from otomycosis cases in Hungary between 2010 and 2011. These strains were identified as A. niger according to conventional morphological methods. Species identification was based on sequencing of part of the calmodulin gene. Our results indicate that instead of A. niger, A. awamori and A. tubingensis are the predominant species that cause ear infections in Southern Hungary. Antifungal susceptibility tests were carried out against four antifungal drugs: amphotericin B, itraconazole, ketoconazole and terbinafine. All isolates were found to exhibit low in vitro MIC values to amphotericin B, terbinafine and itraconazole. However, the examined isolates exhibited high in vitro MIC values to ketoconazole.     

TLR4-dependent GM-CSF protects against lung injury in Gram-negative bacterial pneumonia

Toll-like receptors (TLRs) are required for protective host defense against bacterial pathogens. However, the role of TLRs in regulating lung injury during Gram-negative bacterial pneumonia has not been thoroughly investigated. In this study, experiments were performed to evaluate the role of TLR4 in pulmonary responses against Klebsiella pneumoniae (Kp). Compared with wild-type (WT) (Balb/c) mice, mice with defective TLR4 signaling (TLR4(lps-d) mice) had substantially higher lung bacterial colony-forming units after intratracheal challenge with Kp, which was associated with considerably greater lung permeability and lung cell death. Reduced expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA and protein was noted in lungs and bronchoalveolar lavage fluid of TLR4 mutant mice postintratracheal Kp compared with WT mice, and primary alveolar epithelial cells (AEC) harvested from TLR4(lps-d) mice produced significantly less GM-CSF in vitro in response to heat-killed Kp compared with WT AEC. TLR4(lps-d) AEC underwent significantly more apoptosis in response to heat-killed Kp in vitro, and treatment with GM-CSF protected these cells from apoptosis in response to Kp. Finally, intratracheal administration of GM-CSF in TLR4(lps-d) mice significantly decreased albumin leak, lung cell apoptosis, and bacteremia in Kp-infected mice. Based on these observations, we conclude that TLR4 plays a protective role on lung epithelium during Gram-negative bacterial pneumonia, an effect that is partially mediated by GM-CSF.     

Bi-allelic Truncating Mutations in TANGO2 Cause Infancy-Onset Recurrent Metabolic Crises with Encephalocardiomyopathy

Molecular diagnosis of mitochondrial disorders is challenging because of extreme clinical and genetic heterogeneity. By exome sequencing, we identified three different bi-allelic truncating mutations in TANGO2 in three unrelated individuals with infancy-onset episodic metabolic crises characterized by encephalopathy, hypoglycemia, rhabdomyolysis, arrhythmias, and laboratory findings suggestive of a defect in mitochondrial fatty acid oxidation. Over the course of the disease, all individuals developed global brain atrophy with cognitive impairment and pyramidal signs. TANGO2 (transport and Golgi organization 2) encodes a protein with a putative function in redistribution of Golgi membranes into the endoplasmic reticulum in Drosophila and a mitochondrial localization has been confirmed in mice. Investigation of palmitate-dependent respiration in mutant fibroblasts showed evidence of a functional defect in mitochondrial β-oxidation. Our results establish TANGO2 deficiency as a clinically recognizable cause of pediatric disease with multi-organ involvement.