CCR5 tyrosine sulfation heterogeneity generates cell surface receptor subpopulations with different ligand binding properties.

BACKGROUND: Chemokine receptor tyrosine sulfation plays a key role in the binding of chemokines. It has been suggested that receptor sulfation is heterogeneous, but no experimental evidence has been provided so far. The potent anti-HIV chemokine analog 5P12-RANTES has been proposed to owe its inhibitory activity to a capacity to bind a larger pool of cell surface CCR5 receptors than native chemokines such as CCL5, but the molecular details underlying this phenomenon have not been elucidated. METHODS: We investigated the CCR5 sulfation heterogeneity and the sensitivity of CCR5 ligands to receptor sulfation by performing ELISA assays on synthetic N-terminal sulfopeptides and by performing binding assays on CCR5-expressing cells under conditions that modulate CCR5 sulfation levels. RESULTS: Two commonly used anti-CCR5 monoclonal antibodies with epitopes in the sulfated N-terminal domain of CCR5 show contrasting binding profiles on CCR5 sulfopeptides, incomplete competition with each other for cell surface CCR5, and opposing sensitivities to cellular treatments that affect CCR5 sulfation levels. 5P12-RANTES is less sensitive than native CCL5 to conditions that affect cellular CCR5 sulfation. CONCLUSIONS: CCR5 sulfation is heterogeneous and this affects the binding properties of both native chemokines and antibodies. Enhanced capacity to bind to CCR5 is a component of the inhibitory mechanism of 5P12-RANTES. GENERAL SIGNIFICANCE: We provide the first experimental evidence for sulfation heterogeneity of chemokine receptors and its impact on ligand binding, a phenomenon that is important both for the understanding of chemokine cell biology and for the development of drugs that target chemokine receptors.

Silver nanoparticles coated with dodecanethiol used as fillers in non-cytotoxic and antifungal PBAT surface based on nanocomposites.

In the present study, we report the preparation of antifungal and non-cytotoxic polymer nanocomposites with potential application in biomedical materials. Dodecanethiol-protected silver nanoparticles (AgNPs-DDT) were synthesized by a reduction/precipitation method and dispersed in chloroform to obtain stable colloidal dispersions. PBAT-based nanocomposites containing 0.25, 0.5 and 2 wt% AgNPs-DDT were prepared by casting method. The incorporation of AgNPs-DDT in PBAT matrix resulted in nanocomposites which combine improved mechanical performance and antifungal properties with a non-cytotoxic characteristic.

Gravimetric composition of the rejects coming from the segregation process of the municipal recyclable wastes.

Rejects from selective collection are municipal solid waste (MSW) not used for recycling and are, therefore, destined for the landfill in Brazil. Knowledge of the composition and generation of this waste is important for strategically planning public policies that minimize its generation and its negative environmental impacts. However, this portion of MSW is not very well known. Therefore, the aim of this study was to analyze the rejects from the sorting process of the selective waste collection in the municipality of Blumenau, in the State of Santa Catarina, southern Brazil. The studied rejects came from the largest cooperative in the city, and its composition was sorted into 17 categories of 101 samples over the course of one year, with a total of 3893 kg of analyzed rejects. The waste collected by the selective collection of the municipality was evaluated monthly to determine which part of this quantity became rejects and to determine the composition and seasonality of these rejects. The study found that 30.5% of the waste sorted by the cooperative was rejected. Among these rejects, the presence of materials that could be marketed by the cooperative was verified. Hazardous and/or legally prohibited waste were also identified, as were organics, construction and demolition waste, health care waste, electronics, textiles, footwear, batteries, and bulbs. Seasonal analysis indicated a concerning constant generation of health care waste. Aside from that, there was an increase in the generation of waste from electrical and electronic equipment (EEE) during the Christmas period, when a large part of the population discards their EEE. This information is important for the enforcement of the MSW management structure as well as for educational campaigns aimed at the correct separation of waste that should be sent for selective collection.

Twelve-month consumption of a polyphenol extract from olive (Olea europaea) in a double blind, randomized trial increases serum total osteocalcin levels and improves serum lipid profiles in postmenopausal women with osteopenia.

OBJECTIVES: Osteoporosis is a skeletal disorder characterized by impaired bone turnover and compromised bone strength, thereby predisposing to increased risk of fracture. Preclinical research has shown that compounds produced by the olive tree (Olea europaea), may protect from bone loss, by increasing osteoblast activity at the expense of adipocyte formation. The aim of this exploratory study was to obtain a first insight on the effect of intake of an olive extract on bone turnover in postmenopausal women with decreased bone mass (osteopenia). DESIGN AND SETTING: For that, a double blind, placebo-controlled study was performed in which participants were randomly allocated to either treatment or placebo groups. PARTICIPANTS: 64 osteopenic patients, with a mean bone mineral density (BMD) T-score between -1.5 and -2.5 in the lumbar spine (L2-L4) were included in the study. INTERVENTION AND MEASUREMENTS: PARTICIPANTS received for 12 months daily either 250 mg/day of olive extract and 1000 mg Ca (treatment) or 1000 mg Ca alone (placebo). Primary endpoints consisted of evaluation of bone turnover markers. Secondary endpoints included BMD measurements and blood lipid profiles. RESULTS: After 12 months, the levels of the pro-osteoblastic marker osteocalcin were found to significantly increase in the treatment group as compared to placebo. Simultaneously, BMD decreased in the placebo group, while remaining stable in the treatment group. In addition, improved lipid profiles were observed, with significant decrease in total- and LDL-cholesterol in the treatment group. CONCLUSION: This exploratory study supports preclinical observations and warrants further research by showing that a specific olive polyphenol extract (Bonolive®) affects serum osteocalcin levels and may stabilize lumbar spine BMD. Moreover, the improved blood lipid profiles suggest additional health benefits associated to the intake of the olive polyphenol extract.

Invariant Natural Killer T Cells are Reduced in Hereditary Hemochromatosis Patients.

PURPOSE: Invariant natural killer T (iNKT) cells are CD1d restricted-T cells that react to lipid antigens. iNKT cells were shown to be important in infection, autoimmunity and tumor surveillance. Alterations in the number and function of these cells were described in several pathological conditions including autoimmune and/or liver diseases. CD1d is critical for antigen presentation to iNKT cells, and its expression is increased in liver diseases. The liver is the major organ affected in Hereditary Hemochromatosis (HH), an autosomal recessive disorder caused by excessive iron absorption. Herein, we describe the study of iNKT cells of HH patients. METHODS: Twenty-eight HH patients and 24 control subjects from Santo António Hospital, Porto, were included in this study. Patient's iron biochemical parameters (serum transferrin saturation and ferritin levels) and the liver function marker alanine transaminase (ALT) were determined at the time of study. Peripheral blood iNKT cells were analyzed by flow cytometry using an anti-CD3 antibody and the CD1d tetramer loaded with PBS57. RESULTS: We found a decrease in the percentage and number of circulating iNKT cells from HH patients when compared with control population independently of age. iNKT cell defects were more pronounced in untreated patients, relating with serum ferritin and transferrin saturation levels. No correlation was found with ALT, a marker of active liver dysfunction. CONCLUSIONS: Altogether, our results demonstrate that HH patients have reduced numbers of iNKT cells and that these are influenced by iron overload.

Multiplicity of steady states in a bioreactor during the production of 1,3-propanediol by Clostridium butyricum.

In recent decades, the production of compounds from microorganisms has increased significantly. Glycerol as a source of substrate appears to have great potential, due to its large supply because of the increase in biodiesel production. This paper will discuss the multiplicity of steady states for the production of 1,3-propanediol from glycerol by Clostridium butyricum, employing a model that takes into account inhibition by fermentation products. The theoretical study of bifurcation enabled us to make a qualitative adjustment to the various experimental steady states, using the theoretical steady states obtained from the AUTO2007 program. The theoretical model parameters were varied to fit qualitatively the values of the experimental steady states. In addition, this work is a qualitative study, using experimental steady states that can be used as an initial study for more advanced work on optimizing the production of 1,3-propanediol.

Effects of maternal separation on the dietary preference and behavioral satiety sequence in rats.

This study investigated the effects of maternal separation on the feeding behavior of rats. A maternal separation model was used on postnatal day 1 (PND1), forming the following groups: in the maternal separation (MS) group, pups were separated from their mothers each day from PND1 to PND14, whereas in the control (C) group pups were kept with their mothers. Subgroups were formed to study the effects of light and darkness: control with dark and light exposure, female and male (CF and CM), and maternal separation with dark and light exposure, female and male (SDF, SDM, SLF and SLM). Female rats had higher caloric intake relative to body weight compared with male controls in the dark period only (CF=23.3±0.5 v. CM=18.2±0.7, P<0.001). Macronutrient feeding preferences were observed, with male rats exhibiting higher caloric intake from a protein diet as compared with female rats (CF=4.1±0.7, n=8 v. CM=7.0±0.5, n=8, P<0.05) and satiety development was not interrupted. Female rats had a higher adrenal weight as compared with male rats independently of experimental groups and exhibited a higher concentration of serum triglycerides (n=8, P<0.001). The study indicates possible phenotypic adjustments in the structure of feeding behavior promoted by maternal separation, especially in the dark cycle. The dissociation between the mother's presence and milk intake probably induces adjustments in feeding behavior during adulthood.

Filamentous fungi and media for cellulase production in solid state cultures.

Cellulase production was evaluated in two reference strains (T. reesei Rut-C30 and T. reesei QM9414), two strains isolated from a sugarcane cultivation area (Trichoderma sp. IPT778 and T. harzianum rifai IPT821) and one strain isolated in a program for biodiversity preservation in São Paulo state (Myceliophthora thermophila M77). Solid state cultures were performed using sugarcane bagasse (C), wheat bran (W) and/or soybean bran (S). The highest FPA was 10.6 U/gdm for M77 in SC (10:90) at 80% moisture, which was 4.4 times higher than production in pure W. C was a strong inducer of cellulase production, given that the production level of 6.1 U/gdm in WC (40:60) was 2.5 times higher than in pure W for strain M77; T. reesei Rut-C30 did not respond as strongly with about 1.6-fold surplus production. S advantageously replaced W, as the surplus production on SC (20:80) was 2.3 times relative to WC (20:80) for M77.

Filamentous fungi and media for cellulase production in solid state cultures

Cellulase production was evaluated in two reference strains (T. reesei Rut-C30 and T. reesei QM9414), two strains isolated from a sugarcane cultivation area (Trichoderma sp. IPT778 and T. harzianum rifai IPT821) and one strain isolated in a program for biodiversity preservation in São Paulo state (Myceliophthora thermophila M77). Solid state cultures were performed using sugarcane bagasse (C), wheat bran (W) and/or soybean bran (S). The highest FPA was 10.6 U/gdm for M77 in SC (10:90) at 80% moisture, which was 4.4 times higher than production in pure W. C was a strong inducer of cellulase production, given that the production level of 6.1 U/gdm in WC (40:60) was 2.5 times higher than in pure W for strain M77; T. reesei Rut-C30 did not respond as strongly with about 1.6-fold surplus production. S advantageously replaced W, as the surplus production on SC (20:80) was 2.3 times relative to WC (20:80) for M77.

Efficacy of a novel at-home bleaching technique with carbamide peroxides modified by CPP-ACP and its effect on the microhardness of bleached enamel.

This study was designed to evaluate in vitro the efficacy of a novel at-home bleaching technique using 10% or 16% carbamide peroxide modified by casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and its influence on the microhardness of bleached enamel. A total of 40 bovine incisors were divided into four groups (n=10) according to the bleaching agent used: 10% carbamide peroxide only; a blend of 10% carbamide peroxide and a CPP-ACP paste; 16% carbamide peroxide only; and a blend of 16% carbamide peroxide and a CPP-ACP paste. During the 14-day bleaching regimen, the samples were stored in artificial saliva. The Vickers microhardness and color of the teeth were assessed at baseline (T0) and immediately after the bleaching regimen (T14) using a microhardness tester and a spectrophotometer, respectively. The degree of color change was determined by the Commission Internationale de l'Eclariage (CIE) L*a*b* system (ΔE, ΔL*, Δa*, and Δb*) and Vita shade guide parameters. The data were analyzed by analysis of variance and the Tukey test (p<0.05). The teeth that were bleached with a blend of peroxide (10% or 16%) and the CPP-ACP paste presented increased microhardness values at T14 compared with T0, whereas the samples that were bleached with peroxide only did not show any differences in their microhardness values. All of the bleaching agents were effective at whitening the teeth and did not show a statistically significant difference using the CIEL*a*b* system (ΔE, ΔL*, Δa*, and Δb*) or the Vita shade guide parameters. The use of a CPP-ACP paste with carbamide peroxide bleaching agents increased the bleached enamel's microhardness and did not have an influence on whitening efficacy.

Microbiota of dentinal caries as assessed by reverse-capture checkerboard analysis.

BACKGROUND/AIMS: This study aimed to identify the microbiota of different layers of dentinal caries by using a culture-independent molecular biology approach. METHODS: DNA was extracted from samples taken from 3 distinct layers (superficial, middle and deep) of advanced occlusal caries and analyzed for the presence and relative levels of 28 oral bacterial species/phylotypes using a reverse-capture checkerboard hybridization assay. RESULTS: The mean number of target taxa per layer was 7.7 (± 3.96) in the superficial, 7 (± 3.4) in the middle, and 6.3 (± 3.04) in the deep layer. No statistical significance was observed for these differences (p = 0.36). Overall, the most prevalent taxa in the 3 layers were Atopobium genomospecies C1 (72.5%), Fusobacterium nucleatum (69%), Lactobacillus casei (68%), Veillonella species (55%) and Lactobacillus fermentum (52%). No differences were found in the prevalence rates of the most frequent target species in the 3 layers. The most prevalent taxa found at levels above 10(5) in the advanced front line of deep-dentin caries were Atopobium genomospecies C1, F. nucleatum, Streptococcus mutans, Streptococcus species and Veillonella species. CONCLUSION: The present results revealed that the prevalences of several established or candidate caries pathogens do not differ significantly in the different zones of dentinal caries lesions. The finding that some as-yet-uncharacterized species and novel species were found in high frequencies join other molecular studies to include them in the set of candidate caries pathogens.

Plates made with solid waste from the recycled paper industry.

The results of assays carried out on plates used in the construction industry, manufactured entirely with solid wastes of a recycled paper plant, are presented and compared with the results obtained using agglomerated wood and plywood plates. Previous results had shown that wastes are composed essentially of polymers when these wastes are generated by recycled paper produced with the "waved type II" shavings. These solid wastes were placed in a mold that was heated and pressed with a hydraulic press in order to obtain the plates. The waste-produced plates were submitted to tests for humidity, swelling, water absorption, density, modulus of rupture-static bending, modulus of elasticity and direct screw withdrawal. These same assays had been carried out on two types of commercial wood plates, agglomerated wood and plywood, in order to compare the results with those obtained with the waste plate. Waste plates had similar behavior to the agglomerated wood plate, but it was possible to distinguish greater flexibility in the waste-produced plate and a significant difference in the tests for swelling and water absorption which showed the waste plate had a better performance than the agglomerated wood and plywood plates.

Goat PRND expression pattern suggests its involvement in early sex differentiation.

Expression of the goat prion protein gene locus was assessed by reverse transcriptase-polymerase chain reaction on testes and ovaries at various developmental stages. A weak and stochastic expression of the PRNP and PRNT genes was observed. For PRNT, it is consistent with the detected deletions of two single nucleotides within its open reading frame in ruminant genes. PRND was expressed in both tissues at all stages. Whereas its expression is constant in the ovaries, it increases in testes between 36 and 46 days postcoitum (dpc) and remains high thereafter. In testes, Doppel was found in the nucleus of germinal cells and in the cytoplasm of Leydig cells at 44 dpc. It was detected in the cytoplasm of Leydig cells and of some Sertoli and germinal cells at 62 dpc. In the ovaries, it was observed in the nucleus of germinal cells at 44 dpc and mainly in their cytoplasm at 62 dpc. This expression pattern was shown to parallel that of C-kit and suggests Doppel involvement in early testis differentiation.

Minimally invasive pituitary surgery in a hemorrhagic necrosis of adenoma during pregnancy.

A 29-year-old woman with a prolactin microadenoma was under good control with bromocriptine (BCP) therapy until she became pregnant, when the treatment was stopped. During the third trimester of pregnancy the patient presented a unilateral visual loss and intermittent headaches. BCP was restarted and one week later she developed a complete ophthalmoplegia on the other side. The patient was submitted to an endoscopic transnasal transeptal resection of a hematoma and tumoral tissue in the pituitary region. One month after surgery, at 39 weeks of gestation, the patient spontaneously delivered a healthy girl weighing 3 kg, with an Apgar score of 9 and 10 at five minutes. This is the first case in the literature of a pregnant woman with second and third cranial nerve lesions, submitted to minimally invasive neuroendoscopic transnasal transsphenoidal approach in the third trimester of pregnancy.

[Neuroendoscopic endonasal transseptal approach to the sellar region: study of 30 cases]. / Abordagem neuroendoscopica transnasal transeptal para a região selar: estudo de 30 casos.

An endoscopic endonasal transsphenoidal approach to the sella was performed in 30 patients. The use of endoscope allowed close inspection and differentiation between tumor tissue and glandular remains, resulting in micro dissection of the tumor with maximum preservation of pituitary function. The angled view of the endoscope helps gross total removal of the tumor tissue, from the supra and para sellar extension. The endonasal transseptal endoscopic approach helps from dental problems, and lowers incidence of septal perforation, synechia, and crust formation. However the endoscopic approach has drawbacks. Its decreases the depth of field, needs constant manual control of the endoscope, and requires experience with the endoscope technique.