Morphologic and morphometric analysis of testis of Pseudis limellum (Cope, 1862) (Anura, Hylidae) during the reproductive cycle in the Pantanal, Brazil

The spermatogenesis of Pseudis limellum, from the Southern Pantanal, Brazil, was studied from July 1995 to May 1996, through histological sections of the testis. The cells could be differentiated as: primary spermatogonia, large cells, generally with bilobed nucleus; secondary spermatogonia, smaller cells, with darker cytoplasm, chromatin of radial form; primary and secondary spermatocytes, differentiated according to the different stages of the nucleus during the successive cells divisions. Furthermore, we observed cells in process of morphologic differentiation: rounded spermatids much smaller, with nucleus containing chromatin in compacting process and cytoplasm reduction; elongated spermatids, generally parallel organized in well defined bundles, with the anterior region directed toward the periphery of the seminiferous tubule and the tail directed toward the lumen. Spermatozoa are free in the lumen of the seminiferous tubule. All the cells are organized as cysts, which are supported by a large amount of Sertoli cells. The spermatogenesis in P. limellum is very similar to that of other anurans, but peculiarities were observed regarding the organization of the germ cells, the great amount of free Sertoli cells in the lumen of testis collected in May, and the long cytoplasmatic extensions of the cells bearing pigments and involving the seminiferous tubule. The diameter of the seminiferous tubule (SD) exhibited an annual mean of 251.79 +/- 37.57 microm. Spermatozoa number by seminiferous tubule (SN) exhibited an annual mean of 306.66 +/- 39.83, also with higher and lower values at each month. Variations in SD and SN were not significantly correlated with climatic variables. In this species, reproduction occurs throughout the year in ponds and flooded areas, despite the seasonal climate of the Pantanal. Although males varied in their annual reproductive activity, they were considered potentially reproductive in all months throughout the year.     

Oral Microbiome in Relation to Periodontitis Severity and Systemic Inflammation

Systemic inflammation induced by periodontitis is suggested to be the link between periodontitis and cardiovascular disease. The aim of this work was to explore the oral microbiome in periodontitis in relation to disease severity and systemic inflammation. The saliva and subgingival microbiome from periodontal pocket samples of patients with severe (n = 12) and mild periodontitis (n = 13) were analyzed using metagenomic shotgun sequencing. The taxa and pathways abundances were quantified. The diversity was assessed and the abundances to phenotype associations were performed using ANCOM and linear regression. A panel of inflammatory markers was measured in blood and was associated with taxa abundance. The microbial diversity and species richness did not differ between severe and mild periodontitis in either saliva or periodontal pockets. However, there were significant differences in the microbial composition between severe and mild periodontitis in the subgingival microbiome (i.e., pocket samples) and, in a lower grade, in saliva, and this is positively associated with systemic inflammatory markers. The “red complex” and “cluster B” abundances in periodontal pockets were strongly associated with inflammatory markers interleukin-6 and the white blood cell count. Our data suggest that systemic inflammation in severe periodontitis may be driven by the oral microbiome and may support the indirect (inflammatory) mechanism for the association between periodontitis and cardiovascular disease.     

Dynamic shadow mask technique: a universal tool for nanoscience

A comprehensive instrument, designed for fabricating nanostructures by evaporation through a dynamic shadow mask in ultrahigh vacuum, is described. The versatility and performance of the instrument is demonstrated through a series of examples, allowing for applications that are impossible to achieve with traditional nanopatterning methods. Clean nanostructures or entire devices made of different materials and on various substrates can be fabricated. The technique is compatible with fundamental surface science and can be easily interfaced with other fabrication and characterization techniques.     

Synthesis of BaTiO3 by soft chemistry routes

In this paper we report a comparison concerning the properties of BaTiO₃ (BTO) ceramics obtained by two soft chemical routes, modified Pechini method and thermal decomposition of oxalate-based precursor. XRD data show the formation of single phase BaTiO₃ with tetragonal symmetry when the polymeric citrate-based precursor was annealed at 850 °C, 2 h. In the case of oxalate based-precursor, longer thermal treatment is required to obtain BaTiO₃ free of any secondary phases. For BaTiO₃ powders prepared by modified Pechini method, TEM and SEM investigations revealed the obtaining of uniformly sized particles forming spherical agglomerates inside large, non-uniform and partially sintered aggregates. The powders synthesized via oxalate route show particles of various sizes, with the same tendency of spherical agglomerates formation, but unlike the modified Pechini synthesis, more uniform and smaller aggregates with well-defined hexagonal-like shape were noticed. The relative permittivity values of 6,478 and 5,088 at Curie temperatures of 127 and 130 °C and low dielectric losses (tan δ?=?0.012) at room temperature were obtained for ceramic samples synthesized via Pechini method and oxalate route, respectively.     

Statistical Evaluation of Real-Time PCR Protocols Applied to Quantify Genetically Modified Maize

Real-time PCR (RTi-PCR) is the technique of choice for event-specific quantification of genetically modified organisms (GMOs) by determining the amount of event with respect to a species-specific reference gene. Reference genes can be amplified from the genome extracted from Certified Reference Materials (CRMs) or from ad hoc designed plasmids. In the present study, we statistically evaluate the performance of RTi-PCR protocols for GM maize MON810 event by using both genomic DNA from conventional CRMs and a plasmid containing sequences representative of four maize species-specific reference genes. The significance of simple and interaction effects of several variables included in the experimental design on DNA quantification methods and RTi-PCR were evaluated and discussed. Statistically significant differences on Ct values may have an impact on the GMOs quantification and consequently on the compliance of GM quantification-established legal thresholds. Our results confirm the reliability of the plasmid as alternative calibrant for the calculation of GMOs copy number.     

Combination of freeze drying and molecular inclusion techniques improves the bioaccessibility of microencapsulated anthocyanins from black rice (Oryza sativa L.) and lavender (Lavandula angustifolia L.) essential oils in a model food system

Black rice and lavender are promising sources of bioactives, in terms of anthocyanins and essential oils. Their bioaccessibility were improved by microencapsulation, followed by mixing in order to benefit both from colour and flavour, along with radical scavenging and biological properties. The mixed powder showed a satisfactory anthocyanins of 2.55 mg g⁻¹ DW, leading to a radical scavenging activity of 65.14 mmol g⁻¹ DW. The powder displayed an inhibitory effect against α-glucosidase (~49%) and α-amylase (39%), respectively, with a controlled release in intestinal environment. To further examine the functional properties, the powder was added to a food model system. During storage, a release in anthocyanins and flavonoids were found, leading to an increase in radical scavenging activity. The sensorial analysis showed that supplemented biscuits were appreciated for colour and lavender aroma. The obtained results were promising in tailoring the health benefits of secondary metabolites, underutilised in human’s nutrition due to their low stability and bioavailability.     

Adsorption studies on copper,cadmium, and zinc ion removal from aqueous solution using magnetite/carbon nanocomposites

Magnetite/carbon nanocomposites were tested as adsorbents for removal of metal ions from aqueous solutions. The effect of adsorption parameters such as solutions pH (ranging between 2 and 9), the nature and the quantity of the sorbent (10, 20, 40, and 60 mg), initial concentration of metal ions (10, 30, 50, 100, and 150 mg/L), and temperature (25, 45, and 65°C) was evaluated. The removal efficiency of metal ions depends on solution pH and increases with increasing carbon content, the dose of magnetite/carbon nanocomposites, and the temperature and decrease with initial concentration of the metal ions. The adsorption kinetics was described by pseudo-second-order model, and the equilibrium experimental data were well fitted to the Sips isotherm, yielding a maximum adsorption capacity of 41.11, 76.67, and 48.45 mg/g for copper, cadmium, and zinc, respectively. The thermodynamic parameter Gibbs free energy was determined to be negative, which indicated that the adsorption process is spontaneous. The optimum conditions (1 g/L adsorbent, 25°C, and pH 6) were selected for removal of metal ions from real wastewaters, with good results indicating that investigated nanocomposites could be used for the application in real systems.     

Integration of individual nanoscale structures into devices using dynamic nanostenciling

We succeeded in integrating individual, pre-existing nanostructures into functional devices using ultrahigh vacuum dynamic nanostenciling and show working devices based on single-walled carbon nanotubes, a benchmark nanomaterial, and porphyrin J-aggregates, a "soft" supramolecular nanomaterial. Nanostructures are first located via atomic force microscopy, while device elements are added step by step, with an achieved positional accuracy of 20 nm, using a shadow mask assembly that moves while being exposed to evaporated material. Electronic transport, potentiometry, and scanning Kelvin probe were used for control at any fabrication stage and were available in situ. Such complex fabrication/characterization capabilities, applicable repeatedly, reliably, and nondestructively, pave the way for dynamic nanostenciling instrumentation to establish itself as a viable tool for easy integration and prototyping of fragile nanostructures synthesized through a wide range of processes.     

Establishing In-House Cutoffs of CSF Alzheimer’s Disease Biomarkers for the AT(N) Stratification of the Alzheimer Center Barcelona Cohort

Background: Clinical diagnosis of Alzheimer’s disease (AD) increasingly incorporates CSF biomarkers. However, due to the intrinsic variability of the immunodetection techniques used to measure these biomarkers, establishing in-house cutoffs defining the positivity/negativity of CSF biomarkers is recommended. However, the cutoffs currently published are usually reported by using cross-sectional datasets, not providing evidence about its intrinsic prognostic value when applied to real-world memory clinic cases. Methods: We quantified CSF Aβ1-42, Aβ1-40, t-Tau, and p181Tau with standard INNOTEST^® ELISA and Lumipulse G^® chemiluminescence enzyme immunoassay (CLEIA) performed on the automated Lumipulse G600II. Determination of cutoffs included patients clinically diagnosed with probable Alzheimer’s disease (AD, n = 37) and subjective cognitive decline subjects (SCD, n = 45), cognitively stable for 3 years and with no evidence of brain amyloidosis in 18F-Florbetaben-labeled positron emission tomography (FBB-PET). To compare both methods, a subset of samples for Aβ1-42 (n = 519), t-Tau (n = 399), p181Tau (n = 77), and Aβ1-40 (n = 44) was analyzed. Kappa agreement of single biomarkers and Aβ1-42/Aβ1-40 was evaluated in an independent group of mild cognitive impairment (MCI) and dementia patients (n = 68). Next, established cutoffs were applied to a large real-world cohort of MCI subjects with follow-up data available (n = 647). Results: Cutoff values of Aβ1-42 and t-Tau were higher for CLEIA than for ELISA and similar for p181Tau. Spearman coefficients ranged between 0.81 for Aβ1-40 and 0.96 for p181TAU. Passing–Bablok analysis showed a systematic and proportional difference for all biomarkers but only systematic for Aβ1-40. Bland–Altman analysis showed an average difference between methods in favor of CLEIA. Kappa agreement for single biomarkers was good but lower for the Aβ1-42/Aβ1-40 ratio. Using the calculated cutoffs, we were able to stratify MCI subjects into four AT(N) categories. Kaplan–Meier analyses of AT(N) categories demonstrated gradual and differential dementia conversion rates (p = 9.815⁻²⁷). Multivariate Cox proportional hazard models corroborated these findings, demonstrating that the proposed AT(N) classifier has prognostic value. AT(N) categories are only modestly influenced by other known factors associated with disease progression. Conclusions: We established CLEIA and ELISA internal cutoffs to discriminate AD patients from amyloid-negative SCD individuals. The results obtained by both methods are not interchangeable but show good agreement. CLEIA is a good and faster alternative to manual ELISA for providing AT(N) classification of our patients. AT(N) categories have an impact on disease progression. AT(N) classifiers increase the certainty of the MCI prognosis, which can be instrumental in managing real-world MCI subjects.