Structural insights and antimicrobial synergy of a proto-galectin from the marine sponge Aiolochroia crassa.

In this study, we isolated a novel lectin from the marine sponge Aiolochroia crassa, named AcrL. The lectin showed a preference for glycans containing sialic acid terminal residues, as indicated by the strongest inhibition with fetuin and bovine submaxillary mucin. Primary structure determination by mass spectrometry revealed that AcrL is a galectin with conserved amino acid residues typically involved in carbohydrate binding. Structural modeling indicated that AcrL adopts a typical galectin ß-sandwich motif, featuring two anti-parallel ß-sheets with five strands each. Docking calculations revealed a carbohydrate-binding site composed of a main site, capable of hosting galactopyranosides, and an extended site, facilitating the binding of complex carbohydrates. AcrL inhibited significant biofilm formation against Staphylococcus aureus, S. epidermidis, and Escherichia coli with concentrations ranging from 500 to 15.6 µg.mL-1 for S. aureus, 7.8 µg.mL-1 for S. epidermidis, and 500 µg.mL-1 for E. coli. Furthermore, when combined with different antibiotics, AcrL potentiated their effect against pathogenic bacteria. The antimicrobial mechanism of AcrL was investigated using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The analysis indicates that AcrL induces damage to the bacterial membrane. These findings underscore the discovery of a novel galectin in a basal organism and the comprehensive biochemical characterization conducted in this research, highlighting the potential of AcrL as a novel antibacterial agent and emphasizing its importance in combating bacterial infections.

MiR-9-3 hypermethylation is associated with stages of diabetic retinopathy.

Purpose: To investigate the potential relation between methylation of miR-9-3 and stages of diabetic retinopathy (DR). Additionally, we explored whether miR-9-3 methylation impacts the serum levels of Vascular Endothelial Growth Factor (VEGF). Methods: A cross-sectional study was conducted with 170 participants with type 2 diabetes, including a control group (n = 64) and a diabetes retinopathy group (n = 106), which was further divided into NPDR (n = 58) and PDR (n = 48) subgroups. Epidemiological, clinical, anthropometric, biochemical ELISA assay were analysed. DNA extracted from leukocytes was used to profile miR-9-3 methylation using PCR-MSP. Results: MiR-9-3 hypermethylated profile was higher in the DR group (p < 0.001) and PDR subgroup compared to DM2 control group (p < 0.001). The hypermethylated profile in the PDR subgroup was also higher compared to NPDR subgroup (p < 0.001). There was no difference between DM2 control and NPDR group (p = 0.234). Logistic regression showed that miR-9-3 hypermethylation increases the odds of presenting DR (OR: 2.826; p = 0.002) and PDR (OR: 5.472; p < 0.001). In addition, hypermethylation of miR-9-3 in the DR and NPDR subgroup was associated with higher serum VEGF-A levels (p = 0.012 and p = 0.025, respectively). Conclusion: The methylation profile of the miR-9-3 promoter increases the risk of developing PDR. Higher levels of VEGF-A are associated with miR-9-3 hypermethylated profile in patients in the DR and NPDR stages. Supplementary Information: The online version contains supplementary material available at 10.1007/s40200-024-01411-9.

Metabolomic analysis of retinopathy stages and amputation in type 2 diabetes.

BACKGROUND: Diabetic retinopathy (DR) and limb amputation are frequent complications of diabetes that cannot always be explained by blood glucose control. Metabolomics is a science that is currently being explored in the search for biomarkers or profiles that identify clinical conditions of interest. OBJECTIVE: This study aimed to analyze, using a metabolomic approach, peripheral blood samples from type 2 diabetes mellitus (DM2) individuals, compared with those with diabetic retinopathy and limb amputation. METHODS: The sample consisted of 128 participants, divided into groups: control, DM2 without DR (DM2), non-proliferative DR (DRNP), proliferative DR (DRP), and DM2 amputated (AMP). Metabolites from blood plasma were classified by spectra using nuclear magnetic resonance (NMR), and the metabolic routes of each group using metaboanalyst. RESULTS: We identified that the metabolism of phenylalanine, tyrosine, and tryptophan was discriminant for the DRP group. Histidine biosynthesis, on the other hand, was statistically associated with the AMP group. The results of this work consolidate metabolites such as glutamine and citrulline as discriminating for DRP, and the branched-chain amino acids as important for DR. CONCLUSIONS: The results demonstrate the relationship between the metabolism of ketone bodies, with acetoacetate metabolite being discriminating for the DRP group and histidine being a significant metabolite in the AMP group, when compared to the DM2 group.

Osteoarticular Sporotrichosis of the Knee Caused by Sporothrix brasiliensis: Two Similar Cases with Different Outcomes.

Sporotrichosis is the most frequent subcutaneous or implantation mycosis in Latin America, and its transmission occurs as a result of traumatic inoculation into the skin by organic matter containing the thermodimorphic fungi of the genus Sporothrix. Although cutaneous forms are more common, another important site is the osteoarticular system, whose hematogenous involvement is commonly associated with disseminated forms, especially in people who have an immunosuppressive condition, such as HIV/AIDS, chronic steroid use, and alcohol abuse. We present two cases of osteoarticular sporotrichosis of the knee caused by Sporothrix brasiliensis and followed up at our institution, with different outcomes. In the cases presented here, aging, anatomical sites, comorbidities, subtherapeutic serum levels, low adherence to treatment, and late diagnosis for different reasons may explain the observed outcomes. Early diagnosis of Sporothrix infection is critical in preventing complications, including death. We also highlight the importance of multidisciplinary follow-up and adherence to treatment for a favorable outcome.

Structural study and antimicrobial and wound healing effects of lectin from Solieria filiformis (Kützing) P.W.Gabrielson.

The SfL-1 isoform from the marine red algae Solieria filiformis was produced in recombinant form (rSfL-1) and showed hemagglutinating activity and inhibition similar to native SfL. The analysis of circular dichroism revealed the predominance of ß-strands structures with spectra of ßI-proteins for both lectins, which had Melting Temperature (Tm) between 41 °C and 53 °C. The three-dimensional structure of the rSfL-1 was determined by X-ray crystallography, revealing that it is composed of two ß-barrel domains formed by five antiparallel ß chains linked by a short peptide between the ß-barrels. SfL and rSfL-1 were able to agglutinate strains of Escherichia coli and Staphylococcus aureus and did not show antibacterial activity. However, SfL induced a reduction in E. coli biomass at concentrations from 250 to 125 µg mL-1, whereas rSfL-1 induced reduction in all concentrations tested. Additionally, rSfL-1 at concentrations from 250 to 62.5 µg mL-1, showed a statistically significant reduction in the number of colony-forming units, which was not noticed for SfL. Wound healing assay showed that the treatments with SfL and rSfL-1 act in reducing the inflammatory response and in the activation and proliferation of fibroblasts by a larger and fast deposition of collagen.

Prevalence of staphylococcal toxin in food contaminated by Staphylococcus spp.: Protocol for a systematic review with meta-analysis.

BACKGROUND: Food contamination by Staphylococcus spp. enterotoxigenic strains is quite common and despite underreporting caused by the short duration of clinical symptoms and lack of medical care, staphylococcal food poisoning is one of the most common Foodborne Diseases (FBD) in the world. This study describes a systematic review protocol with meta-analysis on the prevalence and types of staphylococcal enterotoxins in food, and the profile of contaminated foods. METHODS: The research will be conducted through the selection of studies reporting the analysis of staphylococcal enterotoxins in food contaminated by Staphylococcus spp. Searches will happen on the following databases: Medline (OVID), GALE, Science Direct, CAB Direct (CABI), Google Scholar, in addition to manual search in the list of references of articles, directory of theses and dissertations, and countries' health agencies. Reports will be imported into the application Rayyan. Two researchers will independently select studies and extract data, and a third reviewer will solve conflicting decisions. The primary outcome will be the identification of staphylococcal enterotoxins in food, and the secondary outcomes will include staphylococcal enterotoxin types and foods involved. To assess the risk of bias in the studies, the tool developed by the Joanna Briggs Institute (JBI) will be used. For data synthesis, a meta-analysis will be performed. However, in case that is not possible, a narrative synthesis of the most relevant results will be carried out. DISCUSSION: This protocol will serve as the basis for a systematic review that aims to relate the results of existing studies on the staphylococcal enterotoxin prevalence and types in food, and the profile of the contaminated foods. The results will broaden the perception of food safety risks, highlight existing literature gaps, contribute to the study of the epidemiological profile and may guide the allocation of health resources for the development of preventive measures related. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration number: CRD42021258223.

Easy Access to Biomedicine and Knowledge about Medicinal Plants: A Case Study in a Semiarid Region of Brazil.

We aimed to evaluate how proximity to urban areas interferes with the relationship between socioeconomic variables and various aspects of medicinal plant knowledge. The study was conducted in six communities of the Catimbau National Park (PARNA Catimbau) in the state of Pernambuco. Eighty participants were interviewed. The communities were divided into two groups according to their distance from the nearest urban center. Socioeconomic data and information on medicinal plants were collected through semistructured interviews. Subsequently, generalized linear models were generated to verify the influence of the interaction between the variables on medicinal knowledge. We observed that proximity to the urban center influenced the relationship between the level of education and the knowledge of body systems treated by medicinal plants. We concluded that environmental variables can generate a differentiated effect on the influence of socioeconomic factors on one's knowledge of medicinal plants.

Purple grape juice improves performance of recreational runners, but the effect is genotype dependent: a double blind, randomized, controlled trial.

BACKGROUND: We examined the influence of superoxide dismutase 3 (SOD3) Arg213Gly and Peroxisome Proliferator-Activated α-Receptor (PPARα) 7G/C polymorphisms to a single dose of purple grape juice supplementation on time-to-exhaustion running test, redox balance and muscle damage in recreational runners. METHODS: Forty-seven male recreational runners performed a running test until exhaustion after supplementation with grape juice or a control drink. Serum total antioxidant capacity (TAC), malondialdehyde (MDA), plasma nitrite (NO), creatine kinase (CK) and lactate dehydrogenase (LDH) were measured pre and post exercise. Also, polymorphisms were analyzed in DNA extracted from the oral mucosa. RESULTS: Grape juice improved the time-to-exhaustion. When analyzed by genotype, the recreational runners with GG+CG genotypes of the SOD3 gene had greater time-to-exhaustion than the CC genotype, but was no different for the PAPRα gene. A slight difference was noted in TAC, since the CC genotype of the SOD3 gene showed higher TAC values in the post-exercise compared to the baseline and with pre-exercise, but these values did not increase compared to the CG+GG group, respectively. The SOD3 and PPARα genes were similar at all times for the other biochemical variables. CONCLUSION: The ergogenic effect of grape juice was genotype-dependent for SOD3 Arg213Gly. However, biochemical redox balance markers did not explain this difference.

Neuromodulation of Hippocampal-Prefrontal Cortical Synaptic Plasticity and Functional Connectivity: Implications for Neuropsychiatric Disorders.

The hippocampus-prefrontal cortex (HPC-PFC) pathway plays a fundamental role in executive and emotional functions. Neurophysiological studies have begun to unveil the dynamics of HPC-PFC interaction in both immediate demands and long-term adaptations. Disruptions in HPC-PFC functional connectivity can contribute to neuropsychiatric symptoms observed in mental illnesses and neurological conditions, such as schizophrenia, depression, anxiety disorders, and Alzheimer's disease. Given the role in functional and dysfunctional physiology, it is crucial to understand the mechanisms that modulate the dynamics of HPC-PFC communication. Two of the main mechanisms that regulate HPC-PFC interactions are synaptic plasticity and modulatory neurotransmission. Synaptic plasticity can be investigated inducing long-term potentiation or long-term depression, while spontaneous functional connectivity can be inferred by statistical dependencies between the local field potentials of both regions. In turn, several neurotransmitters, such as acetylcholine, dopamine, serotonin, noradrenaline, and endocannabinoids, can regulate the fine-tuning of HPC-PFC connectivity. Despite experimental evidence, the effects of neuromodulation on HPC-PFC neuronal dynamics from cellular to behavioral levels are not fully understood. The current literature lacks a review that focuses on the main neurotransmitter interactions with HPC-PFC activity. Here we reviewed studies showing the effects of the main neurotransmitter systems in long- and short-term HPC-PFC synaptic plasticity. We also looked for the neuromodulatory effects on HPC-PFC oscillatory coordination. Finally, we review the implications of HPC-PFC disruption in synaptic plasticity and functional connectivity on cognition and neuropsychiatric disorders. The comprehensive overview of these impairments could help better understand the role of neuromodulation in HPC-PFC communication and generate insights into the etiology and physiopathology of clinical conditions.

Foliar absorption and field herbicidal studies of atrazine-loaded polymeric nanoparticles.

Nanoparticles loaded with atrazine show weed control efficacy even with lower application doses of the active ingredient. Changes in the mode of action of the herbicide through the nanoformulation are key to understanding the efficiency of post-emergence activity of nanoatrazine. Here, we report the leaf absorption and translocation of nanoatrazine and atrazine employing radiometric techniques and compare their herbicidal effects in greenhouse and field conditions. Compared to the commercial formulation, nanoatrazine showed greater and faster absorption rates in mustard leaves (40% increment in the absorbed herbicide 24 h after application), inducing higher inhibition of photosystem II activity. Assays with fusicoccin-treated leaves indicated that the stomatal uptake of nanoparticles might be involved in the improved activity of nanoatrazine. Nanoencapsulation potentiated the post-emergent herbicidal activity of atrazine and the gain provided by nanoencapsulation was higher in the field compared to greenhouse conditions. Regardless of the dose, nanoatrazine provided two-fold higher weed control in the field compared to commercial atrazine. Thus, the design of this carrier system enables improvements in the performance of the herbicide in the field with less risk of environmental losses of the active ingredients due to faster absorption.

Histoplasmosis tenosynovitis of the forearm and wrist: imaging, surgical and pathologic findings.

Histoplasmosis is a disease endemic to several parts of the world, including South America. It progresses in a mostly asymptomatic and self-limiting manner but has the potential to cause disseminated pulmonary infection, especially in immunocompromised patients. Rare reports from the literature describe musculoskeletal manifestations related to the two varieties of the pathogen that cause histoplasmosis, namely, the capsulatum or duboissi variety. In this report, we describe the case of a previously healthy, middle-aged man, with slowly progressing pain in the right forearm, wrist, and fingers, diagnosed with a case of pathologically confirmed histoplasmosis tenosynovitis. We also describe the imaging, surgical, and histological findings and discuss the differential diagnoses for tenosynovitis, in case of atypical infections.

Oxidative stress does not influence weight loss induced by aerobic training in adults: randomized clinical trials.

BACKGROUND: High levels of oxidative stress promote degradation of the cell membrane impairing cellular function in fat oxidation. However, the influence of oxidative stress on exercise-induced weight-loss has not yet been investigated. Therefore, the aim of this study was to verify the influence of a lipidic peroxidation marker (malondialdehyde, MDA) and antioxidant status (total antioxidant capacity marker, TAC) on the magnitude of weight-loss by aerobic-induced exercise in previously sedentary overweight or obese individuals. METHODS: Seventy-five physically inactive adults were randomized into experimental (N.=58) and control (N.=17) groups, who engaged in a 12-week program of aerobic training walking and/or running (3 to 5 days/week) or stretching (1 day/week), respectively. Body composition (DXA), aerobic capacity (ergospirometric) and blood collections for oxidative stress analysis (MDA and TAC) were determined before and after the experimental protocol. Two-way ANOVA for repeated measures or Friedman's test were used to evaluate differences in time/group interaction. Pearson correlation was used to verify the relationship between the variables of oxidative stress and of body composition. RESULTS: Significant reduction was found in fat body mass of experimental when compared to control group (-1.3±1.9 kg versus -0.3±1.3, P=0.04). Experimental group also altered significantly the total body mass (-1.2±4.7 kg; effect size 0.44), body mass index - BMI (-0.3±1.1 effect size 0.37), fat percentage (1.3±1.6%; effect size 0.50) and lean body mass (0.6±1.5 kg; effect size 0.32).There was increase in MDA of 2.3 µmol/L to 2.7 µmol/L (P=0.00), without changes to TAC (25.6±13.9% to 28.0±10.4%). No correlation was found between these variations in body composition with either the initial values of MDA and TAC or delta variation of these indicators of oxidative stress in response to the training program. CONCLUSIONS: Indicators of oxidative stress (MDA and TAC) does not influence the magnitude of weight-loss induced by aerobic training.

Pro12Ala Polymorphism on the PPARγ2 Gene and Weight Loss After Aerobic Training: A Randomized Controlled Trial.

The objective of this study was to verify the influence of the Pro12Ala polymorphism of the PPARγ2 gene in response of a training program on the body composition. Sixty-nine previously inactive men and women (32.8 ± 8.2 years) were genotyped and underwent a 12-week aerobic (running/walking) training program (3-5 sessions, 40 - 60 min per session, and intensity between the aerobic and anaerobic threshold) (experimental group n = 53) or were part of the control group (n = 16). They were tested for aerobic capacity (ergospirometry), body composition (DXA), abdomen, waist and hip circumferences and nutritional assessment before and 48 h after the experimental protocol. Two-way repeated measures ANOVA test was used to verify possible differences in variables between the experimental vs. control groups or Pro/Pro vs. Pro/Ala groups, and the Chi-squared test was used to verify the distribution of responders and non-responders according to genotype (p < 0.05). Frequencies of 75.5% Pro/Pro (n = 40) and 24.5% Pro/Ala (n = 13) were found, without any occurrence of the recessive homozygote. Body fat reduction was initially confirmed compared to a control group which did not exercise (n = 16; 29.1 ± 8.8 years), so that the exercise group obtained a reduction of -1.3 kg vs. -0.3 kg in the control group (p = 0.03). When they were divided by genotype, there were significant changes in fat mass (-1.3 ± 2.1 kg; p = 0.00), lean mass (0.6 ± 1.5 kg; p = 0.02), fat percentage (-1.3 ± 1.6; p = 0.00), waist circumference (-2.2 ± 2.9 cm; p = 0.00), abdomen circumference (-3.3 ± 3.6 cm; p = 0.00) and hip circumference (-2.7 ± 2.7 cm; p = 0.00) for Pro/Pro genotypes; and fat mass (-1.1 ± 1.7 kg; p = 0.04), fat percentage (-0.9 ± 1.5; p = 0.04), abdomen circumference (-3.9 ± 3.5 cm; p = 0.00) and hip circumference (-1.8 ± 1.8 cm; p = 0.00) for Pro/Ala genotypes, without any group interaction differences. The Chi squared test revealed no differences in the distribution of responders or non-responders according to genotype. It is concluded that an aerobic training program promotes weight loss, but the Pro12Ala polymorphism in the PPARγ2 gene does not influence the variability of aerobic-induced exercise weight loss.

A single dose of purple grape juice improves physical performance and antioxidant activity in runners: a randomized, crossover, double-blind, placebo study.

PURPOSE: To investigate the effects of a single dose of juice on physical performance, oxidative stress, inflammation and muscle damage in runners. METHODS: Fourteen recreational male runners (39 ± 9 years, VO2peak = 55.9 ± 6.5 ml/kg/min) performed two running tests to exhaustion at 80% of VO2max after ingesting grape juice or a placebo drink (10 ml/kg/day) randomly. Blood samples were taken before and 2 h after supplementation and immediately after running to analyze total antioxidant capacity (TAC), malondialdehyde (MDA), alpha-1 acid glycoprotein (A1GPA), high-sensitivity C-reactive protein (hs-CRP), creatine kinase (CK) and lactate dehydrogenase (LDH). RESULTS: The participants ran for an average of 59.2 ± 27.8 min until exhaustion in the placebo group and for 68.4 ± 29.7 min until exhaustion in the grape juice intake group, which was a significantly longer time (p = 0.008). This improvement in physical performance was accompanied by a 43.6% increase in TAC (p = 0.000) at the post-exercise timepoint compared to the level at baseline. MDA, A1GPA, hs-CRP, CK, and LDH did not exhibit changes. In contrast, no significant change in any variable was observed after consuming the placebo drink. CONCLUSION: The single-dose intake of purple grape juice demonstrated an ergogenic effect in recreational runners by increasing run time to exhaustion and increasing antioxidant activity.

Breastfeeding and neurodevelopmental outcomes.

PURPOSE OF REVIEW: Breastfeeding has clear short-term benefits for child survival. Concerning its long-term consequences, it has been reported that subjects who had been breastfed would have a better performance in intelligence tests. In this review, we perused the recently published studies on the association of breastfeeding with developmental outcomes. RECENT FINDINGS: A meta-analysis published in 2015 reported that intelligence quotient (IQ) was 3.44 points (95% confidence interval: 2.30; 4.58) higher among subjects who had been breastfed, and this association was observed even among those studies that controlled for maternal IQ. In the present review, we identified two studies that reported that duration of breastfeeding was positively associated with IQ in childhood, whereas another study reported that cognitive score at 67.9 years of age was higher among those subjects who had been breastfed for at least 6 months. Furthermore, two studies reported that the small gain in IQ was associated with higher school achievement and income in adulthood. Concerning the possible mechanisms for the effect of breastfeeding on development, children who had been breastfed showed greater gray matter volume in the left and right parietal and left temporal lobes and more activation in the right frontal and temporal lobes for perception tasks, whereas for the language task, the activation was higher in the left temporal lobe. Moreover, it has been reported that subcortical gray matter volume mediated the association between breastfeeding and IQ. SUMMARY: The new studies identified in the review reinforce the evidence that breastfeeding has long-term consequences on performance in intelligence tests. In addition, this association between breastfeeding and development has long-term consequences on human capital, increasing earning ability.

Purification, Biochemical Characterization, and Amino Acid Sequence of a Novel Type of Lectin from Aplysia dactylomela Eggs with Antibacterial/Antibiofilm Potential.

A new lectin from Aplysia dactylomela eggs (ADEL) was isolated by affinity chromatography on HCl-activated Sepharose™ media. Hemagglutination caused by ADEL was inhibited by several galactosides, mainly galacturonic acid (Ka = 6.05 × 106 M-1). The primary structure of ADEL consists of 217 residues, including 11 half-cystines involved in five intrachain and one interchain disulfide bond, resulting in a molecular mass of 57,228 ± 2 Da, as determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry. ADEL showed high similarity with lectins isolated from Aplysia eggs, but not with other known lectins, indicating that these lectins could be grouped into a new family of animal lectins. Three glycosylation sites were found in its polypeptide backbone. Data from peptide-N-glycosidase F digestion and MS suggest that all oligosaccharides attached to ADEL are high in mannose. The secondary structure of ADEL is predominantly ß-sheet, and its tertiary structure is sensitive to the presence of ligands, as observed by CD. A 3D structure model of ADEL was created and shows two domains connected by a short loop. Domain A is composed of a flat three-stranded and a curved five-stranded ß-sheet, while domain B presents a flat three-stranded and a curved four-stranded ß-sheet. Molecular docking revealed favorable binding energies for interactions between lectin and galacturonic acid, lactose, galactosamine, and galactose. Moreover, ADEL was able to agglutinate and inhibit biofilm formation of Staphylococcus aureus, suggesting that this lectin may be a potential alternative to conventional use of antimicrobial agents in the treatment of infections caused by Staphylococcal biofilms.

Antiproliferative and ultrastructural effects of phenethylamine derivatives on promastigotes and amastigotes of Leishmania (Leishmania) infantum chagasi.

Leishmania (Leishmania) infantum chagasi is one of the agents that cause visceral leishmaniasis. This disease occurs more frequently in third world countries, such as Brazil. The treatment is arduous, and is dependent on just a few drugs like the antimonial derivatives and amphotericin B. Moreover, these drugs are not only expensive, but they can also cause severe side effects and require long-term treatment. Therefore, it is very important to find new compounds that are effective against leishmaniasis. In the present work we evaluated a new group of synthetic amides against the promastigote and amastigote forms of L. infantum chagasi. The results showed that one of these amides in particular, presented very effective activity against the promastigotes and amastigotes of L. infantum chagasi at low concentrations and it also presented low toxicity for mammal cells, which makes this synthetic amide a promising drug for combating leishmaniasis.

Filling lateral canals: influence of calcium hydroxide paste as a root canal dressing.

CONTEXT: The use of intracanal medication aims to provide antimicrobial activity in the root canal system, leading to a more satisfactory condition for root canal obturation. However, it is important to evaluate whether such medication would affect the sealing-off from the main root canal of ramifications such as lateral canals. AIM: Evaluate the influence of calcium hydroxide [Ca(OH)2] paste on the filling of lateral canals. SETTINGS AND DESIGN: Ex vivo quantitative laboratory study. MATERIALS AND METHODS: Sixty single-rooted teeth with lateral canals created were used, 10 teeth per group. Group A was described respectively as lateral condensation technique (subgroup 1), thermomechanical compaction (subgroup 2), and continuous wave of condensation (subgroup 3). In Group B, subgroups 4, 5, and 6 were filled with Ca(OH)2 for 7 days before filling using the techniques applied in subgroups 1, 2, and 3. Radiographic evaluation was performed to count the number of filled lateral canals in each group. The teeth were cross-sectioned over the lateral canals; thus, 180 specimens were obtained. Each specimen was immersed in a polyester resin. Digital images were obtained and specific software (Image Tool; ) was used to evaluate the number of obturated lateral canals and the filling length in each lateral canal of groups. STATISTICAL ANALYSIS USED: Data were statistically analyzed using SPSS 12.0 for Windows (Descriptive statistical analysis and Newman-Keuls test). RESULTS: A greater number of simulated lateral canals were filled in Group A (91.1%). Group B had less filled lateral canals (33.3%). The sealing of lateral canals was statistically different between Group B (26.15%) and Group A (74.43%). CONCLUSIONS: According to the radiographic analysis and the linear filling measurement, the use of Ca(OH)2 decreased the number and the length of sealed lateral canals.

Molecular characterization and tandem mass spectrometry of the lectin extracted from the seeds of Dioclea sclerocarpa Ducke.

Lectin from the seeds of Dioclea sclerocarpa (DSL) was purified in a single step by affinity chromatography on a Sephadex G-50 column. The primary sequence, as determined by tandem mass spectrometry, revealed a protein with 237 amino acids and 81% of identity with ConA. DSL has a molecular mass of 25,606 Da. The ß and γ chains weigh 12,873 Da and 12,752 Da, respectively. DSL hemagglutinated rabbit erythrocytes (both native and treated with proteolytic enzymes), showing stability even after one hour of exposure to a specific pH range. The hemagglutinating activity of DSL was optimal between pH 6.0 and 8.0, but was inhibited after incubation with D-galactose and D-glucose. The pure protein possesses a molecular mass of 25 kDa by SDS-PAGE and 25,606 Da by mass spectrometry. The secondary structure content was estimated using the software SELCON3. The results indicate that b-sheet secondary structures are predominant in DSL (approximately 42.3% antiparallel b-sheet and 6.7% parallel b-sheet). In addition to the b-sheet, the predicted secondary structure of DSL features 4.1% a-helices, 15.8% turns and 31.3% other contributions. Upon thermal denaturation, evaluated by measuring changes in ellipticity at 218 nm induced by a temperature increase from 20 °C to 98 °C, DSL displayed cooperative sigmoidal behavior with transition midpoint at 84 °C and permitted the observation of two-state model (native and denatured).