Evaluating total nitrogen and phosphorous concentrations in a watershed impacted by diverse anthropic activities in a developing country.

This research aims to identify critical contamination points by nutrients, their possible origin (point and nonpoint sources), their spatial distribution, and possible attenuation by natural and anthropogenic processes. The study area is the Velhas River Basin, located in the Southeast Region of Brazil (17.0°-20.5° S; 43.5°-45.0°W). A historical series of water quality monitoring, land cover map, demographic and agricultural censuses, sewage treatment diagnostics, and local hydrographic networks were used to achieve the objectives. In addition, the regions were divided into incremental areas, enabling individualized analyses of each sub-basin. Descriptive statistics, seasonality, categorized data tests, agglomerative hierarchical cluster analysis, and principal component analysis were used. There was a significant contribution of nutrients in the most important urban agglomeration of the basin, resulting in peak concentrations measured at that place. Although the values were reduced by the mouth (650 km), the percentage of legislation violations remained high. The effects of punctual contamination were intensified by the low percentage of treated sewage in the basin, the absence of adequate treatment technologies to remove nutrients, and the disorderly urbanization. Furthermore, it was estimated that the nutrient load from animal husbandry is approximately 75% of the load from domestic effluents due to the high number of cattle in the basin and the low percentage of forests.

Impact of Compensating Curves on Mastication of Complete Denture Wearers: A Randomised Triple-Blind Clinical Trial.

INTRODUCTION: The application of occlusal concepts in prosthodontics is still under debate.This study assessed the impact of compensating curves on the comminution of complete denture wearers. METHODS: Seven edentulous subjects, aged 64.6 ± 2.0 years, were rehabilitated with new muco-supported complete dentures and tested in two occlusal plane settings: with and without compensating curves. A randomised triple-blind clinical trial was conducted, considering one-week and one-month as adaptation periods for dentures. After each trial, the subjects were crossed over to their respective groups. One-week was also chosen as the washout period, and after that, the subjects were re-examined. The masticatory performance and swallowing threshold were determined while chewing Optocal test food. The multiple sieve method was used for fractionation and granulometry. RESULTS: During the masticatory performance and swallowing threshold estimates, no differences were found between the median particle sizes obtained with the both occlusal plane conditions (P ⟩ 0.05). However, the swallowing threshold improved after one month, resulting in smaller particle sizes. Moreover, the chewing rates for the both test foods were also increased (P ⟨ 0.05). CONCLUSIONS: These findings suggest that the compensating curves did not have an impact on the masticatory function of subjects wearing complete dentures.

A Study of Sponge Symbionts from Different Light Habitats.

The amount of available light plays a key role in the growth and development of microbial communities. In the present study, we tested to what extent sponge-associated prokaryotic communities differed between specimens of the sponge species Cinachyrella kuekenthali and Xestospongia muta collected in dimly lit (caves and at greater depths) versus illuminated (shallow water) habitats. In addition to this, we also collected samples of water, sediment, and another species of Cinachyrella, C. alloclada. Overall, the biotope (sponge host species, sediment, and seawater) proved the major driver of variation in prokaryotic community composition. The light habitat, however, also proved a predictor of compositional variation in prokaryotic communities of both C. kuekenthali and X. muta. We used an exploratory technique based on machine learning to identify features (classes, orders, and OTUs), which distinguished X. muta specimens sampled in dimly lit versus illuminated habitat. We found that the classes Alphaproteobacteria and Rhodothermia and orders Puniceispirillales, Rhodospirillales, Rhodobacterales, and Thalassobaculales were associated with specimens from illuminated, i.e., shallow water habitat, while the classes Dehalococcoidia, Spirochaetia, Entotheonellia, Nitrospiria, Schekmanbacteria, and Poribacteria, and orders Sneathiellales and Actinomarinales were associated with specimens sampled from dimly lit habitat. There was, however, considerable variation within the different light habitats highlighting the importance of other factors in structuring sponge-associated bacterial communities.

Analysis of microbial communities of ocular prostheses and anophthalmic sockets using 16S rRNA gene sequencing.

Ocular prostheses are liable to host microbial adhesion, which can favor conjunctival inflammation. Knowing the microbiota of the ocular prosthesis and anophthalmic socket is important for predicting infection-related pathogens. This study evaluated the microbiota of the prostheses and anophthalmic sockets of six individuals by 16S rRNA sequencing. The microbial abundance and diversity were analyzed using the Operational Taxonomic Units at the genus level. Forty-seven phyla, capturing 1,258 named genera, were recorded in the sample set. In both sites, the most frequent genera were Fusobacterium, Staphylococcus, Prevotella, and Streptococcus. The microbial abundance was higher for the anophthalmic socket than for the prosthesis. Alpha diversity showed no significant differences in bacterial richness or diversity between sites. Although the microbial abundance in the anophthalmic socket was higher, both sites had similar microbiota with high diversity and low dominance among the genera.

Optical fiber thermo-refractometer.

This work presents the implementation of a thermo-refractometer, which integrates the measurement of both refractive index and temperature in a single optical fiber structure. To this purpose, a lossy mode resonance (LMR)-based refractometer is obtained by means of the deposition of a titanium dioxide (TiO2) thin film onto a side-polished (D-shaped) single mode fiber. Measurement and subsequent temperature compensation are achieved by means of a fiber Bragg grating (FBG) inscribed in the core of the D-shaped region. The LMR wavelength shift is monitored in transmission while the FBG (FBG peak at 1533 nm) displacement is observed in reflection. The LMR is sensitive to both the surrounding refractive index (SRI), with a sensitivity of 3725.2 nm/RIU in the 1.3324-1.3479 range, and the temperature (- 0.186 nm/°C); while the FBG is only affected by the temperature (32.6 pm/°C in the 25°C - 45°C range). With these values, it is possible to recover the SRI and temperature variations from the wavelength shifts of the LMR and the FBG, constituting a thermo-refractometer, where it is suppressed the effect of the temperature over the refractometer operation, which could cause errors in the fourth or even third decimal of the measured SRI value.

Nandrolone Supplementation Promotes AMPK Activation and Divergent 18[FDG] PET Brain Connectivity in Adult and Aged Mice.

Decreased anabolic androgen levels are followed by impaired brain energy support and sensing with loss of neural connectivity during physiological aging, providing a neurobiological basis for hormone supplementation. Here, we investigated whether nandrolone decanoate (ND) administration mediates hypothalamic AMPK activation and glucose metabolism, thus affecting metabolic connectivity in brain areas of adult and aged mice. Metabolic interconnected brain areas of rodents can be detected by positron emission tomography using 18FDG-mPET. Albino CF1 mice at 3 and 18 months of age were separated into 4 groups that received daily subcutaneous injections of either ND (15 mg/kg) or vehicle for 15 days. At the in vivo baseline and on the 14th day, brain 18FDG-microPET scans were performed. Hypothalamic pAMPKT172/AMPK protein levels were assessed, and basal mitochondrial respiratory states were evaluated in synaptosomes. A metabolic connectivity network between brain areas was estimated based on 18FDG uptake. We found that ND increased the pAMPKT172/AMPK ratio in both adult and aged mice but increased 18FDG uptake and mitochondrial basal respiration only in adult mice. Furthermore, ND triggered rearrangement in the metabolic connectivity of adult mice and aged mice compared to age-matched controls. Altogether, our findings suggest that ND promotes hypothalamic AMPK activation, and distinct glucose metabolism and metabolic connectivity rearrangements in the brains of adult and aged mice.

Microbiome taxonomic and functional profiles of two domestic sewage treatment systems.

Anaerobic systems for domestic sewage treatment, like septic tanks and anaerobic filters, are used in developing countries due to favorable economic and functional features. The anaerobic filter is used for the treatment of the septic tank effluent, to improve the COD removal efficiency of the system. The microbial composition and diversity of the microbiome from two wastewater treatment systems (factory and rural school) were compared through 16S rRNA gene sequencing using MiSeq 2 × 250 bp Illumina sequencing platform. Additionally, 16S rRNA data were used to predict the functional profile of the microbial communities using PICRUSt2. Results indicated that hydrogenotrophic methanogens, like Methanobacterium, were found in higher abundance in both systems compared to acetotrophic methanogens belonging to Methanosaeta genus. Also, important syntrophic microorganisms (Smithella, Syntrophus, Syntrophobacter) were found in the factory and rural school wastewater treatment systems. Microbial communities were also compared between stages (septic tank and anaerobic filter) of each wastewater treatment stage, revealing that, in the case of the rural school, both microbial communities were quite similar most likely due to hydraulic short-circuit issues. Meanwhile, in the factory, microbial communities from the septic tank and anaerobic filter were different. The school system showed lower COD removal rates (2-30%), which were probably related to a higher abundance of Firmicutes members in addition to the hydraulic short-circuit and low abundance of Chloroflexi members. On the other hand, the fiberglass factory presented higher COD removal rates (60-83%), harboring phyla reported as the core microbiome of anaerobic digesters (Bacteroidetes, Chloroflexi, and Proteobacteria phyla). The knowledge of the structure and composition of wastewater treatment systems may provide support for the improvement of the pollutant removal in anaerobic process.

Epigenetic Markers Associated with Schistosomiasis.

It is important to consider the use of the epigenome as source of complementary data for genome knowledge, which is suitable for the diagnosis of schistosomiasis. Usually, a laboratory diagnosis of schistosomiasis is performed by means of 1. Egg detection in the stool or urine by microscopy remains with limited sensitivity; 2. Immunological screening, in which positivity persists after treatment, and 3. Molecular appraisals prevail over the disadvantages of the currently used methods. In this sense, molecular methodologies are being developed based on epigenetic biomarkers, aiming to improve the diagnosis of the disease and clinical treatment as early as possible to prevent the occurrence of serious liver damage.

Does SARS-Cov-2 invade the brain? Translational lessons from animal models.

The current coronavirus disease (COVID-19) outbreak, caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has raised the possibility of potential neurotropic properties of this virus. Indeed, neurological sequelae of SARS-CoV-2 infection have already been reported and highlight the relevance of considering the neurological impact of coronavirus (CoV) from a translational perspective. Animal models of SARS and Middle East respiratory syndrome, caused by structurally similar CoVs during the 2002 and 2012 epidemics, have provided valuable data on nervous system involvement by CoVs and the potential for central nervous system spread of SARS-CoV-2. One key finding that may unify these pathogens is that all require angiotensin-converting enzyme 2 as a cell entry receptor. The CoV spike glycoprotein, by which SARS-CoV-2 binds to cell membranes, binds angiotensin-converting enzyme 2 with a higher affinity compared with SARS-CoV. The expression of this receptor in neurons and endothelial cells hints that SARS-CoV-2 may have higher neuroinvasive potential compared with previous CoVs. However, it remains to be determined how such invasiveness might contribute to respiratory failure or cause direct neurological damage. Both direct and indirect mechanisms may be of relevance. Clinical heterogeneity potentially driven by differential host immune-mediated responses will require extensive investigation. Development of disease models to anticipate emerging neurological complications and to explore mechanisms of direct or immune-mediated pathogenicity in the short and medium term is therefore of great importance. In this brief review, we describe the current knowledge from models of previous CoV infections and discuss their potential relevance to COVID-19.

Only few benefits from propylene glycol drench in early lactation for cows identified as physiologically imbalanced based on milk spectra analyses.

The main objective of this study was to test the efficiency of a management system combining metabolic clustering of cows based on Fourier-transform mid-infrared (FT-MIR) spectra of milk and targeted treatment of metabolically imbalanced cows with propylene glycol drench. We hypothesized that cows identified in a metabolically imbalanced status during early lactation were associated with subsequent impaired health, reproduction, and production, and that treatment with propylene glycol treatment would improve health, reproduction, and production relatively more in these cows than in control cows. We completed a prospective, randomized controlled trial with 356 early-lactation cows in 2 private dairy herds in Denmark from December 2017 to April 2018. Milk samples of cows were collected before treatment, from 4 to 9 d in milk, and after treatment, from 22 to 27 d in milk. Milk samples were analyzed using FT-MIR spectroscopy. We also measured 4 milk metabolites (ß-hydroxybutyrate, isocitrate, malate, and glutamate) and fat and protein contents. Based on FT-MIR spectra and cluster analyses, cows were clustered into groups of metabolically imbalanced and healthy cows. Within each group, cows were allocated randomly to treatment with propylene glycol (500 mL for 5 d) or no treatment. We analyzed the effect of the treatment on cow-level variables: metabolic cluster, milk metabolites, fat and protein contents, and fat-to-protein ratio at a milk sampling after the treatment. Furthermore, we analyzed daily milk yield, calving to first service interval, and disease occurrence. Results showed only a few effects of propylene glycol treatment and few interactions between treatment and metabolic clusters. We found no significant main effects of propylene glycol treatment in any of these analyses. A negative effect of the imbalanced metabolic cluster was found for the outcome of calving to first service interval for multiparous cows. In conclusion, we found a longer calving to first service interval in metabolically imbalanced cows, but we were not able to demonstrate overall benefits from the applied detection of cows in imbalanced metabolic status in early lactation and follow-up by treatment with propylene glycol.

Effect of access cavity design on gaps and void formation in resin composite restorations following root canal treatment on extracted teeth.

AIM: To evaluate the influence of ultraconservative endodontic access cavities (UEC) on gaps and void formation in resin composite restorations in extracted two-rooted maxillary premolars after root canal treatment. Traditional endodontic access cavities (TEC) were used as a reference for comparison. METHODOLOGY: Two-rooted maxillary premolars were scanned in a micro-computed tomographic (micro-CT) device, matched based on similar anatomical features and allocated into two groups (n = 10) according to the design of the access cavity: TEC or UEC. Teeth were mounted on a mannequin head and a single operator performed the treatment, including endodontic access cavity preparation, root canal detection, preparation, filling and restoration procedures, under magnification. For restorative procedure, a bulk fill flowable layer was applied initially followed by an overlaying of regular composite. After restoration procedures, a new micro-CT scan was performed to check the quality of the coronal restoration by analysing the percentage volume of empty spaces present in the tooth-restoration interface (gaps) and inside the restoration (voids). Data were analysed statistically using Shapiro-Wilk and Student's t tests with a significance level of 5%. RESULTS: All specimens had gaps and voids. There were significant differences between the access cavity designs regarding the formation of voids with significantly more voids associated with the UEC (P < 0.05). Gap formation did not differ between groups (P > 0.05). CONCLUSION: The access cavity design used during root canal treatment interfered with the adaptation of the restorative material. The minimally invasive access cavity design was associated with a significantly greater number of voids within restorations.

Microcosm evaluation of the impact of oil contamination and chemical dispersant addition on bacterial communities and sediment remediation of an estuarine port environment.

AIM: To evaluate the interactive effects of oil contamination and chemical dispersant application on bacterial composition and sediment remediation of an estuarine port environment. METHODS AND RESULTS: A multifactorial controlled microcosm experiment was set up using sediment cores retrieved from an estuarine port area located at Ria de Aveiro lagoon (Aveiro, Portugal). An oil spill with and without chemical dispersant addition was simulated. Sediment oil hydrocarbon concentrations and benthic bacterial community structure were evaluated by GC-MS and 16S rRNA high-throughput sequencing respectively. Although initially (first 10 days) chemical dispersion of oil enhanced the concentrations of the heavier polycyclic aromatic hydrocarbons and of the C22 -C30 alkane group, with time (21 days), no significant differences in hydrocarbon concentrations were detected among treatments. Moreover, no significant changes were detected in the structure of sediment bacterial communities, which mainly consisted of operational taxonomic units related to hydrocarbon-contaminated marine environments. We hypothesize that the environmental background of the sampling site preconditioned the communities' response to additional contamination. CONCLUSION: This experimental microcosm study showed that the chemical dispersion of oil did not influence sediment remediation or bacterial community composition. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study showed that chemical dispersion of oil may not improve the remediation of port sediments. Further studies are needed to investigate the impact of chemical dispersants in combination with bioremediation strategies on the process of sediment remediation in port areas.

The CODATwins Project: The Current Status and Recent Findings of COllaborative Project of Development of Anthropometrical Measures in Twins.

The COllaborative project of Development of Anthropometrical measures in Twins (CODATwins) project is a large international collaborative effort to analyze individual-level phenotype data from twins in multiple cohorts from different environments. The main objective is to study factors that modify genetic and environmental variation of height, body mass index (BMI, kg/m2) and size at birth, and additionally to address other research questions such as long-term consequences of birth size. The project started in 2013 and is open to all twin projects in the world having height and weight measures on twins with information on zygosity. Thus far, 54 twin projects from 24 countries have provided individual-level data. The CODATwins database includes 489,981 twin individuals (228,635 complete twin pairs). Since many twin cohorts have collected longitudinal data, there is a total of 1,049,785 height and weight observations. For many cohorts, we also have information on birth weight and length, own smoking behavior and own or parental education. We found that the heritability estimates of height and BMI systematically changed from infancy to old age. Remarkably, only minor differences in the heritability estimates were found across cultural-geographic regions, measurement time and birth cohort for height and BMI. In addition to genetic epidemiological studies, we looked at associations of height and BMI with education, birth weight and smoking status. Within-family analyses examined differences within same-sex and opposite-sex dizygotic twins in birth size and later development. The CODATwins project demonstrates the feasibility and value of international collaboration to address gene-by-exposure interactions that require large sample sizes and address the effects of different exposures across time, geographical regions and socioeconomic status.

Laundry wastewater and domestic sewage pilot-scale anaerobic treatment: Microbial community resilience regarding sulfide production.

In this study, the removal of anionic surfactant Linear Alkylbenzene Sulfonate (LAS) from laundry wastewater was evaluated in co-digestion with domestic sewage, using a pilot-scale Expanded Granular Sludge Bed reactor. Surfactant influent concentration was enhanced from 5 ±â€¯3 mg LAS L-1 (stage I) to 19 ±â€¯10 mg LAS L-1 (stage II) and 36 ±â€¯19 mg LAS L-1 (stage III) throughout reactor operation. Sulfide levels higher than 20 mg L-1 influenced LAS removal efficiency, which decreased from 71% to 55% and 32% in stage I, II and III, respectively. Acclimation of microbial population was verified and higher relative abundance of the genera similar to Cytophaga, Bacteroides, Syntrophus and Syntrophobacter in the early stages (adaptation and stage I) was replaced by higher relative abundance of the genera Anaerophaga, Nitrosovibrio, Sulfurovum and Desulfovibrio in the last stages (stage II and III).

Morphological changes of the internal carotid artery: prevalence and characteristics. A clinical and ultrasonographic study in a series of 19 804 patients over 25 years old.

BACKGROUND AND PURPOSE: Morphological changes of the internal carotid arteries (McICA) are frequently found during cervical ultrasound studies. However, the etiology of McICA remains controversial. During this study, the prevalence and demographic characteristics of McICA, such as kinking, coiling or looping identified by Doppler ultrasound, were analysed and its relationship with vascular risk factors and stroke was assessed. METHODS: A retrospective study was performed by analysing 19 804 patients who were subjected to cervical ultrasonographic study between January 2000 and June 2012. The data were statistically analysed with SPSS® 20 and a multivariate logistic regression was performed. Statistical significance was accepted for P < 0.05 and 95% confidence intervals (CIs) were used. RESULTS: Morphological changes of the internal carotid arteries were present in 2678 patients (13.5%) and were unilateral in 61.6% of these cases. Carotid kinking was found in 80% of the patients, coiling in 16% and looping in 1%. In multivariate analysis, the presence of McICA was related to older groups (1.04; 95% CI, 1.04-1.05; P < 0.01), female gender (1.78; 95% CI, 1.64-1.94; P < 0.01), patients with hyperlipidemia (1.28; 95% CI, 1.17-1.40; P < 0.01), carotid thickness (1.22; 95% CI, 1.13-1.33; P < 0.01) and cardiac or cardioembolic disease (1.11; 95% CI, 1.01-1.21; P = 0.02). The results of this study indicate that kinking in the carotid artery was associated with ipsilateral cerebral ischemic events (1.43; 95% CI, 1.040-1.958; P < 0.05). CONCLUSION: Morphological changes of the internal carotid arteries were associated with aging, female gender and patients with hyperlipidemia, hypertension, diabetes and heart disease. Kinking was associated with ipsilateral cerebral ischemia.

High irisin levels in overweight/obese children and its positive correlation with metabolic profile, blood pressure, and endothelial progenitor cells.

BACKGROUND AND AIMS: Irisin is involved in the compensatory mechanisms for metabolic regulation and appears to be associated with glucose homeostasis and lipid profile. However, it's possible implications on obesity-associated cardiometabolic complications have not been completely elucidated. This study aimed to investigate the association between irisin level and anthropometric data, metabolic parameters, blood pressure, and endothelial progenitor cells (EPCs) level among children with overweight/obesity. METHODS AND RESULTS: This study included 24 children with overweight/obesity (9 girls and 15 boys) and 63 children with normal weight (25 girls and 38 boys). The anthropometric data, blood pressure, blood biochemistry, EPCs and irisin levels were evaluated. Children with overweight/obesity had significantly higher circulating irisin and EPCs levels than those with normal weight (P < 0.001). Additionally, we found that irisin level was positively correlated with BMI (rho = 0.407), waist circumference (rho = 0.449), triglycerides (rho = .334), glucose (rho = 0.226), insulin (rho = 0.533), HOMA (rho = 0.545), and negatively correlated with HDL cholesterol level (rho = -0.218). Importantly, we also found that irisin levels were significantly correlated with systolic (rho = 0.420), diastolic (rho = 0.331) blood pressure and circulating EPCs level (rho = 0.391). CONCLUSION: Our study provides evidence that overweight/obese children had elevated circulating levels of both irisin and EPCs and address the gap in the literature with regard to the understanding of the implications of irisin on obesity-related cardiometabolic complications among these children and also highlight the possible involvement of irisin regulation on insulin resistance and endothelial function in childhood overweight and obesity.

Model for the separate collection of packaging waste in Portuguese low-performing recycling regions.

Separate collection of packaging waste (glass; plastic/metals; paper/cardboard), is currently a widespread practice throughout Europe. It enables the recovery of good quality recyclable materials. However, separate collection performance are quite heterogeneous, with some countries reaching higher levels than others. In the present work, separate collection of packaging waste has been evaluated in a low-performance recycling region in Portugal in order to investigate which factors are most affecting the performance in bring-bank collection system. The variability of separate collection yields (kg per inhabitant per year) among 42 municipalities was scrutinized for the year 2015 against possible explanatory factors. A total of 14 possible explanatory factors were analysed, falling into two groups: socio-economic/demographic and waste collection service related. Regression models were built in an attempt to evaluate the individual effect of each factor on separate collection yields and predict changes on the collection yields by acting on those factors. The best model obtained is capable to explain 73% of the variation found in the separate collection yields. The model includes the following statistically significant indicators affecting the success of separate collection yields: i) inhabitants per bring-bank; ii) relative accessibility to bring-banks; iii) degree of urbanization; iv) number of school years attended; and v) area. The model presented in this work was developed specifically for the bring-bank system, has an explanatory power and quantifies the impact of each factor on separate collection yields. It can therefore be used as a support tool by local and regional waste management authorities in the definition of future strategies to increase collection of recyclables of good quality and to achieve national and regional targets.

Metal loads and biomarker suite responses in a tropical carnivorous fish indicative of anthropogenic impacts in a Southeastern Brazilian lagoon.

Tropical coastal lagoons are highly productive environments exhibiting high biodiversity. However, the use of these ecosystems by local communities is of concern, since this generally leads to environmental degradation. The Imboassica coastal lagoon, located in Macaé city, in Northern Rio de Janeiro, is an important ecosystem in the state, however, already displaying signs of anthropogenic impacts. Carnivorous fish Hoplias malabaricus specimens were sampled from this impacted site, as well as from a reference area. Fish from Imboassica Lagoon presented lower condition factor, lower cholinesterase activity, and higher percentage of erythrocyte micronuclei when compared to fish from the reference site. Metals in fish from Imboassica Lagoon were always higher than Encantada Lagoon, with some seasonal differences, where some metals were higher in the rainy season compared to the dry season in muscle tissue, with the exception of Cu, Fe, Sr, and Zn; and in the liver, except for Ba, Cd, Cr, Ni, and Sr. Cr and Mn in the edible muscle portion of the fish were higher than the limits established by Brazilian and International legislations as permissible for human consumption, thus leading to concerns regarding public health risks for the local population that use fish as their main protein source.

Biophysical and biological properties of small linear peptides derived from crotamine, a cationic antimicrobial/antitumoral toxin with cell penetrating and cargo delivery abilities.

Crotamine is a natural polypeptide from snake venom which delivers nucleic acid molecules into cells, besides having pronounced affinity for negatively charged membranes and antifungal activity. We previously demonstrated that crotamine derived short linear peptides were not very effective as antifungal, although the non-structured recombinant crotamine was overridingly more potent compared to the native structured crotamine. Aiming to identify the features necessary for the antifungal activity of crotamine, two linear short peptides, each comprising half of the total positively charged amino acid residues of the full-length crotamine were evaluated here to show that these linear peptides keep the ability to interact with lipid membrane model systems with different phospholipid compositions, even after forming complexes with DNA. Interestingly, the presence of cysteine residues in the structure of these linear peptides highly influenced the antifungal activity, which was not associated to the lipid membrane lytic activity. In addition to the importance of the positive charges, the crucial role of cysteine residues was noticed for these linear analogs of crotamine, although the tridimensional structure and lipid membrane lytic activity observed only for native crotamine was not essential for the antifungal activity. As these peptides still keep the ability to form complexes with DNA molecules with no prejudice to their ability to bind to lipid membranes, they may be potentially advantageous as membrane translocation vector, as they do not show lipid membrane lytic activity and may harbor or not antifungal activity, by keeping or not the semi-essential amino acid cysteine in their sequence.

-318C/T polymorphism of the CTLA-4 gene is an independent risk factor for RBC alloimmunization among sickle cell disease patients.

Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) molecule is expressed on T-lymphocyte membrane and negatively influences the antigen-presenting process. Reduced expression of CTLA-4 due to gene polymorphisms is associated with increased risk of autoimmune disorders, whose physiopathology is similar to that of post-transfusion red blood cell (RBC) alloimmunization. Our goal was to evaluate if polymorphisms of CTLA-4 gene that affect protein expression are associated with RBC alloimmunization. This was a case-control study in which 134 sickle cell disease (SCD) patients and 253 non-SCD patients were included. All patients were genotyped for the polymorphisms 49A/G and -318C/T of CTLA-4 gene. The genotype frequency of -318C/T differed significantly between alloimmunized and nonalloimmunized SCD patients, irrespective of clinical confounders (p = .016). SCD patients heterozygous for -318T allele presented higher risk of alloantibody development (OR: 5.4, CI: 1.15-25.6). In conclusion, the polymorphism -318C/T of CTLA-4 gene is associated with RBC alloimmunization among SCD patients. This highlights the role played by CTLA-4 on post-transfusion alloantibody development.