Morphological and ultrastructural changes in fungal agents after LASER application.

BACKGROUND: Onychomycosis is a fungal nail infection of difficult treatment due to the fungal survival capacity and reduced number of effective therapies. The present study aimed to isolate fungal agents that cause onychomycosis in immunocompetent patients and evaluate how LASER treatments affect the growth and ultrastructure of isolates. METHODS: In total, 21 patients with positive direct microscopic examination (DME) for onychomycosis had nail samples collected for cultivation and phenotypic identification of microorganisms. From these patients, 12 underwent LASER treatment, divided in Group 1 (n = 5) treated with Nd: YAG 1,064 nm, and Group 2 (n = 7) treated with Nd: YAG 1,064 nm + Er: YAG 2,940 nm + topical isoconazole. Transmission Electron Microscopy (TEM) was performed to evaluate ultrastructural changes after treatment. RESULTS: DME, cultivation, and phenotypic identification showed that the most identified fungus was Trichophyton rubrum spp. After LASER therapy, sample cultivation showed alterations in the fungal morphology with reduction of hyphae, conidia, and reproductive structures. Alterations in fungal cell wall structure, cytoplasm density, and organelles were observed by TEM. CONCLUSION: LASER irradiation causes changes in the fungal cells, especially in the number of hyphae and the presence of conidia. In addition, it affects fungal growth and reproduction capacity, which interferes with their infection ability and virulence.

Long-term Ecological Research: Chasing fashions or being prepared for fashion changes?

Long-term-ecological-research (LTER) faces many challenges, including the difficulty of obtaining long-term funding, changes in research questions and sampling designs, keeping researchers collecting standardized data for many years, impediments to interactions with local people, and the difficulty of integrating the needs of local decision makers with "big science". These issues result in a lack of universally accepted guidelines as to how research should be done and integrated among LTER sites. Here we discuss how the RAPELD (standardized field infrastructure system), can help deal with these issues as a complementary technique in LTER studies, allowing comparisons across landscapes and ecosystems and reducing sampling costs. RAPELD uses local surveys to understand broad spatial and temporal patterns while enhancing decision-making and training of researchers, local indigenous groups and traditional communities. Sampling of ecological data can be carried out by different researchers through standardized protocols, resulting in spatial data that can be used to answer temporal questions, and allow new questions to be investigated. Results can also be integrated into existing biodiversity networks. Integrated systems are the most efficient way to save resources, maximize results, and accumulate information that can be used in the face of the unknown unknowns upon which our future depends.

A New Species of Cosmocercoides (Ascaridida: Cosmocercidae) Parasitic in Tree Frogs from Southern Amazonia.

Cosmocercoides meridionalis n. sp. is described from the large and small intestines of 5 species of tree frogs belonging to the families Hylidae and Phyllomedusidae collected from 2 localities in Southern Amazonia. The new species is allocated to the genus Cosmocercoides Wilkie, 1930, due to the presence of papillae in rosettes, which are complex caudal papillae surrounded by punctuations. Cosmocercoides meridionalis n. sp. differs from its congeners by a combination of the following characters: the absence of the gubernaculum, the total length of the female (twice the size of the male), and the presence of 26 rosette papillae with a unique arrangement and distribution pattern: 11 pre-cloacal pairs, 1 ad-cloacal pair, and 1 post-cloacal pair. This is the first species of the genus described from the Amazonia region.

Use of synthetic oligonucleotides for determination of HTLV-1 proviral load by real-time PCR: a helpful alternative approach in the clinical management.

AIMS: To evaluate the potential use of synthetic oligonucleotides as a standard curve for proviral load (PVL) of human T-cell leukaemia virus type 1 (HTLV-1) quantification in peripheral blood mononuclear cells (PBMC) of HTLV-1-infected individuals by quantitative real-time polymerase chain reaction (qPCR) analysis. METHODS AND RESULTS: Synthetic oligonucleotides based on HTLV-1 genome were customized to use as a standard curve. Twelve anti-HTLV-1-positive samples with known HTLV-1 PVL, previously quantified by qPCR assay using TARL-2 cells as a conventional standard curve, were submitted to the new protocol. The proviral quantification levels had a high concordance with qPCR results using a conventional standard curve. The results demonstrate that the conventional standard curve can be replaced by a synthetic standard curve due to its ability to quantification based on the linearity and qPCR efficiency and similar results with a validated qPCR assay using a conventional standard curve. CONCLUSIONS: Synthetic oligonucleotides standard curves could be a very useful tool on HTLV-1 diagnosis and absolute HTLV-1 PVL quantification. SIGNIFICANCE AND IMPACT OF THE STUDY: HTLV-1 PVL determination using synthetic oligonucleotides standard curve by qPCR could be a helpful alternative for the laboratories that monitor infected patients as an important prognostic factor in HTLV-1-associated diseases progression. Also, it can decrease costs and overcome the biological limitations of the plasmid curve.

Environmental structure affects taxonomic diversity but not functional structure of understory birds in the southwestern Brazilian Amazon

Changes in environmental characteristics can affect habitat use by birds, impacting the number of individuals, number of species, and changing species composition and functional structure of assemblages. Metrics that evaluate the functional structure of biological assemblages constitute a complementary tool to the traditional taxonomic approach, because they quantify the differences between species by means of functional traits. We assessed the effect of environmental characteristics on the taxonomic diversity (species richness, species composition and number of individuals) and functional structure (functional richness, functional evenness, Rao's functional diversity, and community-weighted mean traits) of bird assemblages in northern Mato Grosso state, in the southwestern Brazilian Amazon. We sampled birds in 32 plots. At each plot birds were captured using mist nets, and eight environmental variables were measured: canopy openness, leaf litter, elevation, number of trees in three classes of DBH, soil clay content, and distance to nearest stream. To evaluate functional structure, we measured seven morphological traits from individuals of each bird species. Habitat variables had a significant effect on taxonomic diversity. However, the general functional structure was not affected. Elevation and distance to nearest stream were the main variables driving changes in taxonomic diversity and had a minor effect on functional richness. The other metrics of functional structure were not significantly affected by the set of environmental variables. Our results suggest that the sampled bird assemblages exhibit some functional tolerance (redundancy) to small-scale environmental variation, implying certain resilience to ecosystem modification. (AU)

Biodiversity studies through public-private partnership (PPP): The case of Fazenda São Nicolau in the northwest of Mato Grosso.

Peugeot-ONF Forest Carbon Sink Project, located at Fazenda São Nicolau (FSN) in Cotriguaçu Municipality, is a public-private initiative and has been supported by several French and Brazilian institutions. Its Research Program includes studies aiming Biodiversity Conservation, Ecology, Carbon Dynamics, Silvicultural Techniques. FSN comprises a high richness of species with more than 20 new species of beetles (Histeridae, Scarabaeidae e Melolonthidae) described among them one endemic genus. A quick overview of the SuperFamily Scarabaeoidea within the FSN area registered 260 species which nearly 100 species might be new to Science. One new species of fish as well as research with amphibian toxins as tools to treat cancer and malaria are also done with material collected at the FSN. Ecology of the decomposer´s fauna at reforestation sites showed that after 10 years, such places were more similar to native forest sites than pastures or abandoned pastures. Finally large mammals studies indicated that many large mammals use FSN as a refuge. We stress the importance to keep the FSN natural habitats linked with the Juruena National Park. The FSN contour is under heavy deforestation which will increase the species loss and turn FSN in one of the few major natural vegetation fragments.

Respiratory and upper limb function as outcome measures in ambulant and non-ambulant subjects with Duchenne muscular dystrophy: A prospective multicentre study.

The field of translational research in Duchenne muscular dystrophy (DMD) has been transformed in the last decade by a number of therapeutic targets, mostly studied in ambulant patients. A paucity of studies focus on measures that capture the non-ambulant stage of the disease, and the transition between the ambulant and non-ambulant phase. In this prospective natural history study, we report the results of a comprehensive assessment of respiratory, upper limb function and upper limb muscle strength in a group of 89 DMD boys followed in 3 European countries, 81 receiving corticosteroids, spanning a wide age range (5-18 years) and functional abilities, from ambulant (n = 60) to non-ambulant (n = 29). Respiratory decline could be detected in the early ambulatory phase using Peak Expiratory Flow percentage predicted (PEF%), despite glucocorticoid use (mean annual decline: 4.08, 95% CI [-7.44,-0.72], p = 0.02 in ambulant; 4.81, 95% CI [-6.79,-2.82], p < 0.001 in non-ambulant). FVC% captured disease progression in non-ambulant DMD subjects, with an annual loss of 5.47% (95% CI [-6.48,-4.45], p < 0.001). Upper limb function measured with the Performance of Upper Limb (PUL 1.2) showed an annual loss of 4.13 points (95% CI [-4.79,3.47], p < 0.001) in the non-ambulant cohort. Measures of upper limb strength (MyoGrip and MyoPinch) showed a continuous decline independent of the ambulatory status, when reported as percentage predicted (grip force -5.51%, 95% CI [-6.54,-4.48], p < 0.001 in ambulant and a slower decline -2.86%; 95% CI -3.29,-2.43, p < 0.001, in non-ambulant; pinch force: -2.66%, 95% CI [-3.82,-1.51], p < 0.001 in ambulant and -2.23%, 95% CI [-2.92,-1.53], p < 0.001 in non-ambulant). Furthermore, we also explored the novel concept of a composite endpoint by combining respiratory, upper limb function and force domains: we were able to identify clear clinical progression in patients in whom an isolated measurement of only one of these domains failed to appreciate the yearly change. Our study contributes to the field of natural history of DMD, linking the ambulant and non-ambulant phases of the disease, and suggests that composite scores should be explored further.

Cubic PdNP-based air-breathing cathodes integrated in glucose hybrid biofuel cells.

Cubic Pd nanoparticles (PdNPs) were synthesized using ascorbic acid as a reducing agent and were evaluated for the catalytic oxygen reduction reaction. PdNPs were confined with multiwalled carbon nanotube (MWCNT) dispersions to form black suspensions and these inks were dropcast onto glassy carbon electrodes. Different nanoparticle sizes were synthesized and investigated upon oxygen reduction capacities (onset potential and electrocatalytic current densities) under O2 saturated conditions at varying pH values. Strong evidence of O2 diffusion limitation was demonstrated. In order to overcome oxygen concentration and diffusion limitations in solution, we used a gas diffusion layer to create a PdNP-based air-breathing cathode, which delivered -1.5 mA cm(-2) at 0.0 V with an onset potential of 0.4 V. This air-breathing cathode was combined with a specially designed phenanthrolinequinone/glucose dehydrogenase-based anode to form a complete glucose/O2 hybrid bio-fuel cell providing an open circuit voltage of 0.554 V and delivering a maximal power output of 184 ± 21 µW cm(-2) at 0.19 V and pH 7.0.

Isolated new onset 'atypical' optic neuritis in the NMO clinic: serum antibodies, prognoses and diagnoses at follow-up.

Severe, recurrent or bilateral optic neuritis (ON) often falls within the neuromyelitis optica spectrum disorders (NMOSD), but the diagnosis can be particularly challenging and has important treatment implications. We report the features, course and outcomes of patients presenting with atypical ON when isolated at onset. We retrospectively analyzed 69 sequential patients referred to a single UK NMO center with isolated ON at onset. Aquaporin-4 antibody (AQP4-Ab) assessment was performed in all patients and IgG1 myelin-oligodenrocyte glycoprotein (MOG-Ab) in AQP4-Ab(neg) patients. 37 AQP4-Ab positive (AQP4-Ab(pos)) and 32 AQP4-Ab negative (AQP4-Ab(neg)) patients (8 with MOG-Ab) were identified. The AQP4-Ab(neg) group included heterogeneous diagnoses: multiple sclerosis (MS), NMO, relapsing isolated ON (RION), monophasic isolated ON and relapsing acute disseminated encephalomyelitis (ADEM)-like syndromes. Compared to AQP4-Ab(neg) patients, AQP4-Ab(pos) patients had a worse residual visual outcome from first attack (median VFSS 4 vs. 0, p = 0.010) and at last assessment (median VFSS 5 versus 2, p = 0.005). However, AQP4-Ab(neg) patients with RION also had poor visual outcome. Up to 35% of AQP4-Ab(neg) patients developed a LETM and two developed low positivity for AQP4-Ab over time. Eight AQP4-Ab(neg) patients (25%) were MOG-Ab positive, covering a range of phenotypes excluding MS; the first ON attack was often bilateral and most had relapsing disease with a poor final visual outcome [VFSS 4, range (0-6)]. In conlcusion, AQP4-Ab positivity is confirmed as a predictor of poor visual outcome but AQP4-Ab(neg) RION also had a poor visual outcome. Of those without AQP4-Ab, 25% had MOG-Ab and another 25% developed MS; thus, MOG-Ab is associated with AQP4-Ab(neg) non-MS ON.

Respiratory rate estimation from the oscillometric waveform obtained from a non-invasive cuff-based blood pressure device.

The presence of respiratory activity in the electrocardiogram (ECG), the pulse oximeter's photoplethysmo-graphic and continuous arterial blood pressure signals is a well-documented phenomenon. In this paper, we demonstrate that such information is also present in the oscillometric signal acquired from automatic non-invasive blood pressure monitors, and may be used to estimate the vital sign respiratory rate (RR). We propose a novel method that combines the information from the two respiratory-induced variations (frequency and amplitude) via frequency analysis to both estimate RR and eliminate estimations considered to be unreliable because of poor signal quality. The method was evaluated using data acquired from 40 subjects containing ECG, respiration and blood pressure waveforms, the latter acquired using an in-house built blood pressure device that is able to connect to a mobile phone. Results demonstrated a good RR estimation accuracy of our method when compared to the reference values extracted from the reference respiration waveforms (mean absolute error of 2.69 breaths/min), which is comparable to existing methods in the literature that extract RR from other physiological signals. The proposed method has been implemented in Java on the Android device for use in an mHealth platform.

Polyhydroxyalkanoates production by a mixed photosynthetic consortium of bacteria and algae.

For the first time, a mixed photosynthetic culture (MPC) consisting of a consortium of bacteria and algae was investigated for its capacity to accumulate polyhydroxyalkanoates (PHA). The culture was subjected to a feast and famine regime in an illuminated environment without supplying oxygen or any other electron acceptor. The MPC accumulated PHA during the feast phase and consumed it in the famine phase, where the PHA consumption was made possible due to oxygen production by algae. The internal cycling of carbohydrates was also observed, which was likely linked to bacterial glycogen being used as an additional source of energy for acetate uptake during the feast phase, and restored in the famine phase via PHA degradation. The MPC reached a PHA content of 20%, with a PHA storage yield per acetate similar to aerobic systems, opening up the possibility of a new sunlight-driven PHA production process without the need for aeration.

A pilot study assessing the effect of prolonged administration of high daily doses of vitamin D on the clinical course of vitiligo and psoriasis.

Autoimmunity has been associated with vitamin D deficiency and resistance, with gene polymorphisms related to vitamin D metabolism frequently described in affected patients. High doses of vitamin D3 may conceivably compensate for inherited resistance to its biological effects. This study aimed to assess the efficacy and safety of prolonged high-dose vitamin D3 treatment of patients with psoriasis and vitiligo. Nine patients with psoriasis and 16 patients with vitiligo received vitamin D3 35,000 IU once daily for six months in association with a low-calcium diet (avoiding dairy products and calcium-enriched foods like oat, rice or soya "milk") and hydration (minimum 2.5 L daily). All psoriasis patients were scored according to "Psoriasis Area and Severity Index" (PASI) at baseline and after treatment. Evaluation of clinical response of vitiligo patients required a quartile grading scale. All patients presented low vitamin D status (serum 25(OH)D3 ≤ 30 ng/mL) at baseline. After treatment 25(OH)D3 levels significantly increased (from 14.9 ± 7.4 to 106.3 ± 31.9 ng/mL and from 18.4 ± 8.9 to 132.5 ± 37.0 ng/mL) and PTH levels significantly decreased (from 57.8 ± 16.7 to 28.9 ± 8.2 pg/mL and from 55.3 ± 25.0 to 25.4 ± 10.7 pg/mL) in patients with psoriasis and vitiligo respectively. PTH and 25(OH)D3 serum concentrations correlated inversely. The PASI score significantly improved in all nine patients with psoriasis. Fourteen of 16 patients with vitiligo had 25-75% repigmentation. Serum urea, creatinine and calcium (total and ionized) did not change and urinary calcium excretion increased within the normal range. High-dose vitamin D3 therapy may be effective and safe for vitiligo and psoriasis patients.

A robust flow-cytometric protocol for assessing growth rate of hatchery-reared barramundi Lates calcarifer larvae.

In this study, a flow-cytometric cell cycle analysis method to assess instantaneous growth rate of whole larvae of the Australian barramundi Lates calcarifer was developed and validated. High-resolution DNA measurements of either fresh, frozen or RNAlater-preserved larvae (gap0-gap1, G(0) -G(1), coefficient of variation (c.v.) < 3, 4 and 5%, respectively) enabled the deconvolution of the DNA histogram and assignment of the proportion of nuclei into cell cycle compartments G(0) -G(1), S (DNA synthesis) and G(2) -M (Gap2-Mitosis). This technique can be also used for individual fish tissues such as brain, liver, fin and muscle. For the first time, the combined proportion of replicating nuclei (into S and G(2) -M phases) of whole fish larvae and absolute growth rate in length (mm day(-1)) has been correlated in commercial aquaculture conditions. Fast growing L. calcarifer larvae had an overall hyperplasia advantage as indicated by a greater proportion of cells in the S+G(2) -M phase compared with slow growing larvae, which might explain the increasing differences in size during culture. In a fasting trial, larvae ceased growth while maintaining the constant initial rates of cell division throughout a 6 day period. For a highly fed fast growing control group, cell division rates significantly increased after day 4. Flow-cytometric cell cycle analysis of whole fish larvae may provide fish biologists and aquaculturists with a better understanding of how cell division rates influence early growth in natural and artificial environments.

Discovery of GSK1070916, a potent and selective inhibitor of Aurora B/C kinase.

The Aurora kinases play critical roles in the regulation of mitosis and are frequently overexpressed or amplified in human tumors. Selective inhibitors may provide a new therapy for the treatment of tumors with Aurora kinase amplification. Herein we describe our lead optimization efforts within a 7-azaindole-based series culminating in the identification of GSK1070916 (17k). Key to the advancement of the series was the introduction of a 2-aryl group containing a basic amine onto the azaindole leading to significantly improved cellular activity. Compound 17k is a potent and selective ATP-competitive inhibitor of Aurora B and C with K(i)* values of 0.38 +/- 0.29 and 1.5 +/- 0.4 nM, respectively, and is >250-fold selective over Aurora A. Biochemical characterization revealed that compound 17k has an extremely slow dissociation half-life from Aurora B (>480 min), distinguishing it from clinical compounds 1 and 2. In vitro treatment of A549 human lung cancer cells with compound 17k results in a potent antiproliferative effect (EC(50) = 7 nM). Intraperitoneal administration of 17k in mice bearing human tumor xenografts leads to inhibition of histone H3 phosphorylation at serine 10 in human colon cancer (Colo205) and tumor regression in human leukemia (HL-60). Compound 17k is being progressed to human clinical trials.

Effect of specific exercise training on bone mineral density in women with postmenopausal osteopenia or osteoporosis.

AIM: To analyse the effect of a specific program of weight training exercise with closed kinetic chain in bone mineral density in postmenopausal women with osteopenia or osteoporosis. METHODS: A total of 59 postmenopausal women with osteoporosis or osteopenia were included in this prospective study. Subjects were divided into two groups: the study group (SG, n = 30; 57.5 + or - 5.1 years) and the control group (CG, n = 29; 56.6 + or - 4.6 years). In the study group was applied a weight exercise protocol (longitudinal forces in closed kinetic chain) during 12 months, whereas in the control group no weight exercise protocol was applied. Bone mineral density at the lumbar spine and hip was assessed at baseline and at the end of follow-up by dual energy X-ray absorptiometry. RESULTS. Although no significant intragroup differences were found, patients in SG showed a 1.17% increase in the lumbar spine whereas in CG a 2.26% decrease in bone density was detected. CONCLUSION: This protocol of weight training exercise did not significantly improve bone mineral density in postmenopausal women with osteopenia or osteoporosis, but in comparison to the control group, the results showed the importance of practising the specific exercise program for maintenance of bone health in postmenopausal women.

Application of ozonation to reduce biological sludge production in an industrial wastewater treatment plant.

Biosolids production in the activated sludge process generates an additional cost to wastewater treatment plants due to the growing requirements for sludge treatment and disposal. This work focuses on the application of ozonation to reduce sludge production in an industrial wastewater treatment plant. The results show that ozonation was able to promote cell wall rupture, releasing intracellular matter into the liquid medium. This effect was observed by the increase in concentrations of DNA (1.14 to 7.83 mg/L) and proteins (0.5 to 45.602 mg/L) in the liquid phase, when ozonation was applied during 10 min, using 30 mg/L of ozone. Reduction of sludge production was assessed by calculating the observed sludge yield coefficient (Y) in bench-scale continuous experiments conducted with varying proportions of ozonated sludge in the recycle stream and recycle ratios. Reduction of sludge production ranged from 14 to 39%, depending on the experimental conditions. The best result in terms of sludge excess reduction was achieved when 20% of the recycle sludge was ozonated and the recycle ratio was 0.67.