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This study characterized the binding mechanisms of the lectin cMoL (from Moringa oleifera seeds) to carbohydrates using spectroscopy and molecular dynamics (MD). The interaction with carbohydrates was studied by evaluating lectin fluorescence emission after titration with glucose or galactose (2.0-11 mM). The Stern-Volmer constant (Ksv), binding constant (Ka), Gibbs free energy (∆G), and Hill coefficient were calculated. After the urea-induced denaturation of cMoL, evaluations were performed using fluorescence spectroscopy, circular dichroism (CD), and hemagglutinating activity (HA) evaluations. The MD simulations were performed using the Amber 20 package. The decrease in Ksv revealed that cMoL interacts with carbohydrates via a static mechanism. The cMoL bound carbohydrates spontaneously (ΔG < 0) and presented a Ka on the order of 10 2 , with high selectivity for glucose. Protein-ligand complexes were stabilized by hydrogen bonds and hydrophobic interactions. The Hill parameter (h~2) indicated that the binding occurs through the cMoL dimer. The loss of HA at urea concentrations at which the fluorescence and CD spectra indicated protein monomerization confirmed these results. The MD simulations revealed that glucose bound to the large cavity formed between the monomers. In conclusion, the biotechnological application of cMoL lectin requires specific methods or media to improve its dimeric protein structure.

Objective: To digitally evaluate the three-dimensional (3D) remodelling of FGG used to treat RT2 gingival recessions and lack of keratinized tissue on mandibular incisor teeth., Methods: Data from 45 patients included in a previous multicentric RCT were analyzed. Silicone impressions were taken before (baseline) and 3, 6 and 12 months after standardized FGG placement. Casts were scanned and images were superimposed, using digital software, to obtain measurements of estimated soft tissue thickness (eTT; 1, 3, and 5 mm apical to baseline gingival margin). In addition, soft tissue volume (STV) and creeping attachment (CA) were assessed., Results: All patients exhibited postoperative eTT and STV increases, at all time points. The greatest mean thickness gain was observed at eTT3 (1.0 ± 0.4 mm) at 12 months. At 12 months, STV was 52.3 ± 21.1 mm 3 , without relevant changes compared to the 3- and 6-month follow-up. CA, which was observed as early as six months postoperatively, was evident in ∼85 % of teeth at 12 months., Conclusions: Application of FGG was an effective phenotype modification therapy, as shown by the significantly increased tissue thickness postoperatively. Despite the use of FGG technique not aiming for root coverage, digital 3D assessment documented the early and frequent postoperative occurrence of CA, which helped improve recession treatment outcomes., Clinical Significance: The use of 3D assessment methodology allows precise identification of the tissue gain obtained with FGG, which, regardless of technique, results in predictable phenotype modification and frequent occurrence of creeping attachment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Germ cell tumors (GCTs) constitute diverse neoplasms arising in the gonads or extragonadal locations. Testicular GCTs (TGCTs) are the predominant solid tumors in adolescents and young men. Despite cisplatin serving as the primary therapeutic intervention for TGCTs, 10‑20% of patients with advanced disease demonstrate resistance to cisplatin‑based chemotherapy, and epithelial‑mesenchymal transition (EMT) is a potential contributor to this resistance. EMT is regulated by various factors, including the snail family transcriptional repressor 2 ( SLUG ) transcriptional factor, and, to the best of our knowledge, remains unexplored within TGCTs. Therefore, the present study investigated the EMT transcription factor SLUG in TGCTs. In silico analyses were performed to investigate the expression of EMT markers in TGCTs. In addition, a cisplatin‑resistant model for TGCTs was developed using the NTERA‑2 cell line, and a mouse model was also established. Subsequently, EMT was assessed both in vitro and in vivo within the cisplatin‑resistant models using quantitative PCR and western blot analyses. The results of the in silico analysis showed that the different histologies exhibited distinct expression profiles for EMT markers. Seminomas exhibited a lower expression of EMT markers, whereas embryonal carcinomas and mixed GCT demonstrated high expression. Notably, patients with lower SLUG expression had longer median progression‑free survival (46.4 months vs. 28.0 months, P=0.022). In the in vitro analysis, EMT‑associated genes [fibronectin; vimentin ( VIM ); actin, α2, smooth muscle; collagen type I α1; transforming growth factor‑β1; and SLUG ] were upregulated in the cisplatin‑resistant NTERA‑2 (NTERA‑2R) cell line after 72 h of cisplatin treatment. Consistent with this finding, the NTERA‑2R mouse model demonstrated a significant upregulation in the expression levels of VIM and SLUG. In conclusion, the present findings suggested that SLUG may serve a crucial role in connecting EMT with the development of cisplatin resistance, and targeting SLUG may be a putative therapeutic strategy to mitigate cisplatin resistance.

Introduction: Occupational therapists and physiotherapists aim to promote health, prevent various diseases and help people in their rehabilitation processes. So far, there is a paucity of understanding of the big picture of how the new paradigm of planetary health (PH) is connected to the education and practice of these professionals., Methods: This research aimed to address this gap by investigating and deploying a bibliometric analysis to elucidate the pivotal role of occupational therapists and physiotherapists in addressing PH challenges. The ultimate goal is to construct a comprehensive framework crosschecking the bibliometric analysis and the collection of 10 case studies selected by experts to outline how best practices in occupational therapy and physiotherapy, related to the three pillars of sustainability and the UN's Sustainable Development Goals (SDGs), can contribute to increasing PH., Results: The bibliometric analysis revealed four major research strands: 1) enhancing patient care and quality of life; 2) integrating sustainability in health care and rehabilitation; 3) professional development and clinical competence; and 4) evidence-based practice and quality improvement. Moreover, further temporal analysis revealed how the topic evolved, from advancing evidence-based practice and clinical effectiveness, exploring the strengthening of health care and person-centred practices, to connecting the topic to aspects also predicted by the SDGs, such as integrating environmental and climate concerns in therapy and addressing psychological and self-care impacts on health. The case studies confirmed this trend, and a framework of PH in occupational therapy and physiotherapy through the lens of the SDGs was developed to support future research and practitioners in advancing this research field., Conclusions: Occupational therapists and physiotherapists are essential players in public health and can integrate sustainability at every level of practice, from using resources during therapy sessions to advocating for more sustainable lifestyles., (© 2024 The Authors. Australian Occupational Therapy Journal published by John Wiley & Sons Australia, Ltd on behalf of Occupational Therapy Australia.)

This study aimed to analyze the toxic effects of Roundup Transorb® on the endangered Neotropical annual killifish Austrolebias charrua through the assessment of molecular and biochemical biomarkers. The fish were collected in temporary ponds and exposed to environmentally realistic concentrations of the herbicide (5 mg.L -1 for 96 h). The production of ROS, lipid peroxidation, DNA damage, and membrane fluidity were evaluated in the blood cells by flow cytometry. The mRNA expression of the antioxidant-related genes sod2, cat, gstα, atp1a1, gclc, and ucp1 across the brain, liver, and gills was quantified. The acute exposure of annual killifish to Roundup significantly increased ROS production, lipid peroxidation, and DNA damage in their erythrocytes. Likewise, Roundup Transorb® decreased membrane fluidity in the blood cells of the exposed fish. Gene expression analysis revealed that Roundup exposure alters the relative expression of genes associated with oxidative stress and antioxidant defense. Our results give rise to new insights into adaptive mechanisms of A. charrua in response to Roundup. Since Brazilian annual killifishes strongly risk extinction, this study paves the way for developing novel biotechnologies applied to environmental monitoring and aquatic toxicology assessment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Despite the benefits of breastfeeding, early weaning is a reality, so less than 50% of children worldwide and in Brazil are on exclusive breastfeeding in the sixth month of life. A strategy to counteract this scenario is breastfeeding counseling. This study aims to verify the effectiveness of individualized counseling by nurses trained in breastfeeding counseling, on the duration of exclusive breastfeeding, compared to standard care., Methods: Multicenter, randomized, parallel, and open clinical trial, with primiparous women aged over 18 years, hospitalized in rooming-in wards at participating centers and hemodynamically stable, aware, and oriented, who had a single-fetus pregnancy and gave birth, regardless of the type of delivery, with live child, gestational age of 37 to 42 weeks and birth weight greater than 2500 g. The women will be initially approached in rooming-in wards and, upon consent to participate in the study, will be allocated through randomization by blocks composed of eight participants in two groups: intervention and control. The randomization lists will be organized by a central without involvement with the study, which will manage the allocation groups and be prepared in the Randon® program. Women allocated to the intervention group will receive breastfeeding counseling by trained nurses, and those in the control group will receive standard care at the center participating in the study., Discussion: The results can contribute to breastfeeding by evidencing possible exclusivity and duration of the counseling trained nurses provide., Trial Registration: REBEC RBR-4w9v5rq (UTN: U1111-1284-3559) ( https://ensaiosclinicos.gov.br/rg/RBR-4w9v5rq ). Posted on March 20, 2023., (© 2023. The Author(s).)

Ethnopharmacological Relevance: Moringa oleifera Lam. leaves infusion and powder are widely used by population due the nutritional and medicinal potentials, however data regarding safety of use are still inconclusive, leading to prohibition of this plant in some countries., Aim of the Study: The present work investigated the nutritional and phytochemical composition, acute and 28-day repeated dose toxicity, and genotoxicity of M. oleifera leaves infusion and powder., Materials and Methods: For nutritional characterization of leaf powder, it was determined: humidity; mineral residue (ash); total lipid, protein, carbohydrate, and crude fiber contents; and total caloric value. Phytochemical composition was determined by high performance liquid chromatography (HPLC). The acute toxicity assay used Swiss female albino mice and oral administration in a single dose at 2000 and 5000 mg/kg of infusion or powder. The 28-day repeated dose toxicity assay employed female and male mice, with oral administration of infusion or powder at the doses 250, 500 and 1000 mg/kg. The animals were evaluated for body weight, water and feed consumption, biochemical and hematological parameters, and histology of the liver, spleen, and kidneys. In vivo genotoxicity and mutagenicity (2000 mg/kg) were evaluated by the comet assay and the micronucleus test, respectively., Results: Nutritional characterization confirmed that M. oleifera leaves are rich in proteins, carbohydrates, lipids, minerals, and fiber. HPLC indicated the presence of flavonoids and cinnamic derivatives as major polyphenols. Acute toxicity did not reveal alterations in weight gain and water and feed consumptions and no change in biochemical, hematological, and histological parameters. Behavior alterations was observed in the first 2 h after administration at 5000 mg/kg in both treatments. Infusion did not present toxicity when administered for 28 days. Conversely, the powder at 500 and 1000 mg/kg promoted liver and kidney damages observed through biochemical parameters and histopathology. Genotoxicity and mutagenicity were not detected at 2000 mg/kg., Conclusions: The present study reveals that M. oleifera leaves are an important source of polyphenols and nutrients. Indiscriminate use of both infusion and crude leaf powder above 2000 mg/kg and powder at 500 and 1000 mg/kg are not recommended. Chronic toxicological studies and establishment of preparation protocols are suggested aiming to guarantee the safety in the use of M. oleifera leaves as nutraceutical by population., (Copyright © 2022. Published by Elsevier B.V.)

Ataxia is defined as a lack of coordination of voluntary movement, caused by a variety of factors. Ataxia can be classified by the age at onset and type (chronic or acute). The causative lesions involve the cerebellum and cerebellar connections. The correct, appropriate use of neuroimaging, particularly magnetic resonance imaging, can make the diagnosis relatively straightforward and facilitate implementation of the appropriate clinical management. The purpose of this pictorial essay is to describe the imaging findings of ataxia, based on cases obtained from the archives of a tertiary care hospital, with a review of the most important findings. We also discuss and review the imaging aspects of neoplastic diseases, malformations, degenerative diseases, and hereditary diseases related to ataxia.

Ataxia is defined as a lack of coordination of voluntary movement, caused by a variety of factors. Ataxia can be classified by the age at onset and type (chronic or acute). The causative lesions involve the cerebellum and cerebellar connections. The correct, appropriate use of neuroimaging, particularly magnetic resonance imaging, can make the diagnosis relatively accurate and facilitate implementation of the appropriate clinical management. The purpose of this pictorial essay is to describe the imaging findings of ataxia, based on cases obtained from the archives of a tertiary care hospital, with a review of the most important findings. We also review and discuss the imaging aspects of infectious, toxic, vascular, and inflammatory diseases.

Knowledge on chronic myelomonocytic leukemia (CMML) patients from Argentina and Brazil is limited. Our series of 280 patients depicted an older age at diagnosis (median 72 years old), 26% of aberrant karyotypes, and a prevalence of myelodysplastic (60%) and CMML-0 subtypes (56%). The median overall survival (OS) was 48.2 months for patients in CMML-0 (Ref.), 24.7 months for those in CMML-1 (HR = 2.0, p = 0.001), and 8.8 months for patients in CMML-2 (HR = 4.6, p < 0.001). In the CMML-0 category, median OS were different between myelodysplastic and myeloproliferative subtypes (63.7 vs 21.2 months, p < 0.001); however, no differences were observed within CMML-1 and CMML-2 subtypes (24.7 vs 23.7 months, p = 0.540, and 9.1 vs 8.2 months, p = 0.160). The prognostic impact of 24 variables and 7 prognostic systems was adjusted to the WHO 2016 after validating their usefulness. Multivariate analysis were performed, and the final model revealed Hb ≥ 8 -< 10g/dL (HR 1.7), Hb < 8g/dL (HR 2.8), poor karyotypes (HR 2.1), WHO 2016-CMML-1 (HR 2.1), and CMML-2 (HR 3.5) as independent adverse clinical parameters in our cohort with a borderline influence of platelets count < 50 × 10 9 /L (HR 1.4). We could validate several scoring systems, the WHO 2016 proposal and its prognostic capability, along with accessible covariates, on predicting the outcome in our series of CMML patients from Latin America.

Egletes viscosa is a plant with therapeutic value due to its antibacterial, antinociceptive and gastroprotective properties. This study aimed to purify, characterize, and evaluate the cytotoxicity of a lectin (EgviL) from the floral capitula of E. viscosa. The lectin was isolated from saline extract through precipitation with ammonium sulfate followed by Sephadex G-75 chromatography. The molecular mass and isoelectric point (pI) of EgviL were determined as well as its temperature and pH stability. Physical-chemical parameters of interaction between EgviL and carbohydrates were investigated by fluorescence quenching and 1 H nuclear magnetic resonance (NMR). Cytotoxicity was investigated against human peripheral blood mononuclear cells (PBMCs) and neoplastic cells. EgviL (28.8 kDa, pI 5.4) showed hemagglutinating activity stable towards heating until 60 °C and at the pH range 5.0-7.0. This lectin is able to interact through hydrophobic and electrostatic bonds with galactose and glucose, respectively. EgviL reduced the viability of PBMCs only at the highest concentration tested (100 μg/mL) while was toxic to Jurkat E6-1 cells with IC 50 of 24.1 μg/mL,inducing apoptosis. In summary, EgviL is a galactose/glucose-binding protein with acidic character, stable to heating and with cytotoxic effect on leukemic cells., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives: To radiographically evaluate the effect of the adjunctive use of tenting screws (TS) for primary horizontal guided bone regeneration (GBR)., Materials & Methods: Twenty-eight patients in need of staged bone augmentation were consecutively treated in a private practice. A xenogenic particulate bone substitute material (DBBM) and a resorbable collagen membrane were used in all patients. Subjects were divided into two groups: control (conventional GBR; n = 22) and test (tenting screws in conjunction with GBR - TS; n = 22). CBCT images were obtained before augmentation and after 6-8 months. CBCTs were superimposed, and linear horizontal measurements were performed. Alveolar ridge width (RW) and ridge width change (RWchange) were assessed at 1, 3, 5, and 7 mm below the bone crest., Results: Forty-four sites in 28 patients were evaluated. No differences between the groups were detected for RW at baseline (TS: 5.87 ± 2.41; control: 5.36 ± 1.65). Regarding RWchange, TS promoted an additional effect at 1 and 3 mm below the crest compared to control (p < .05; RWchange-1 TS: 3.72 ± 2.46; control: 1.25 ± 3.05; RWchange-3 TS: 3.98 ± 2.53; control: 2.50 ± 2.02). The final RW was greater in group TS compared to the control group at the 1, 3, and 5 mm level (p < .05)., Conclusions: The use of tenting screws exerted a positive effect on staged GBR with a greater final RW at the 3 mm level. In addition, GBR in conjunction with TS was able to provide consistent bone augmentation at lingual/palatal sites., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Purpose: To investigate the presence of the variants of lysyl oxygenase ( LOX ) and superoxide dismutase 1 ( SOD1 ) genes in Brazilian patients with advanced keratoconus., Methods: Donor genomic DNA extracted from blood samples was screened for 5'UTR, exonic LOX, and SOD1 variants in a subset of 26 patients presenting with advanced keratoconus (KISA > 1000% and I-S > 2.0) by Sanger sequencing. The impact of non-synonymous amino acid changes was evaluated by SIFT, PMUT, and PolyPhen algorithms. The Mutation Taster tool was used to evaluate the potential impact of formation of new donor and acceptor splice sites in the promoter region of affected volunteers carrying sequence variants. A 7-base SOD1 deletion (IVS2 + 50del7bp) previously associated with keratoconus was screened in 140 patients presenting classical keratoconus by gel fragment analysis, and positive samples were sequenced for confirmation., Results: We found an unreported missense variant in LOX exon 6 in one heterozygous patient, leading to substitution of proline with threonine at residue 392 (p. Thr392Pro) of LOX protein sequence. This mutation was predicted to be potentially damaging to LOX protein. Another LOX variant, Arg158Gln, was also detected in another patient but predicted to be non-pathogenic. Two additional new polymorphisms in LOX 5'UTR region (-116C > T and -58C > T) were found in two patients presenting with advanced keratoconus and were predicted to modulate or create donor/acceptor splice sites in LOX transcripts. Additionally, SOD1 deletion was detected in one patient presenting with severe keratoconus, not in control samples., Conclusion: We described three novel LOX polymorphisms identified for the first time in Brazilian patients with advanced keratoconus, as well as a previously described SOD1 deletion strongly associated with keratoconus. A possible role of these variants in modulating transcript levels in the cornea of affected individual requires further investigation., Competing Interests: There are no conflicts of interest., (Copyright © 2020 Gadelha et al.)

Background and Objectives: Antibodies of unknown specificity (AUS) are frequently identified in the pre-transfusion testing. These antibodies can be insignificant or potentially cause post-transfusion haemolysis. Information about the prevalence of clinically relevant AUS is still lacking. Our aim was to predict the potential clinical relevance of AUS using the monocyte monolayer assay (MMA) and to identify the clinical and laboratorial determinants of AUS' significance., Materials and Methods: Antibodies of unknown specificity identified at a single institution from 2015-2017 were evaluated through MMA. A monocyte index (MI) of more than 5% was predictive of potential post-transfusion haemolysis., Results: Thirty-two patients with AUS were included in the study. Of the studied AUS, 37·5% (12/32) presented with a monocyte index (MI) more than 5%. In the group of significant AUS, 41·7% of the patients presented with sickle cell disease (SCD) and the AUS were associated with Rh antibodies in 75% of the cases. In the group of insignificant AUS, only 10% of the patients had SCD and the association with Rh antibodies was detected in 20% of the cases. The presence of Rh antibodies was independently associated with the AUS clinical relevance (P = 0·012)., Conclusion: More than one-third of the AUS are potentially clinically relevant, and the association with Rh antibodies is predictive of AUS relevance. Services must honour AUS in the pre-transfusion process in order to ensure transfusion safety., (© 2019 International Society of Blood Transfusion.)

The aim of this case report is to describe the diagnosis and treatment of a patient with necrotizing ulcerative gingivitis. An 18-year-old man with no systemic problems reported with chief complaints of gingival bleeding during toothbrushing and spontaneous pain. Clinical examination revealed significant plaque accumulation on the surfaces of all teeth as well as papillary necrosis involving mainly the anterior teeth. Treatment included an initial phase of supragingival plaque and calculus removal along with at-home use of 0.12% chlorhexidine gluconate mouthrinse twice a day for 30 days. After the initial phase, subgingival scaling was performed, and regular oral hygiene methods were resumed by the patient. After active therapy was completed, a periodontal maintenance regimen was established, and the patient was recalled for periodontal maintenance therapy. Follow-up occurred weekly throughout treatment, monthly for the first 6 months posttreatment, and 2-3 times a year during the periodontal maintenance therapy. Clinical results after 10 years showed that this approach controlled the acute phase and maintained the patient's periodontal health over time.

Aim: The aim of this study is to report root coverage outcomes in single deep gingival recessions (GR) following a proposed decision-making algorithm., Materials and Methods: A retrospective, practice-based study included single deep (≥5 mm) Miller Class II and III defects. The step-by-step decision-making algorithm led to a choice among three different flap designs (coronally advanced flap (CAF), double papilla envelope flap (DPE) or modified lateral sliding flap (LSF)) used with a connective tissue graft. Recession depth (RD) at 6 months follow-up and the corresponding root coverage (RC) were the primary outcomes assessed., Results: Sixteen GR defects were included, with baseline RD of 6.7 ± 1.8 mm. Six months postoperatively, RD was significantly reduced to 1.2 ± 0.8 mm ( p < 0.05). Mean RC was 81.7 ± 13.0%, without significant differences between Miller Class II (87.1 ± 9.2%; n =9) and Class III (74.6 ± 14.5%; n =7) GRs ( p =0.07). Postoperatively, keratinized tissue width increase was greater for LSF (3.5 ± 1.1) and DPE (4.2 ± 1.4 mm) than for CAF (1.9 ± 0.9 mm)., Conclusions: Following the proposed decision-making algorithm, root coverage outcomes for GR defects ≥5 mm were comparable to outcomes reported for shallow defects. Prospective clinical trials are needed to validate the proposed approach and techniques., Practical Implications: The proposed algorithm allows the clinician to select the appropriate surgical technique for treatment of single deep gingival recessions with good predictability.

The aim of this study was to evaluate the effect of a lower dose of parathyroid hormone- PTH (1-34) on osteogenic potential of bone healing around titanium implants inserted into the tibia of rats. A blind parallel study was conducted in 45 adult male Wistar rats. Each rat received one titanium implant (4.5 x 2.2 mm) and was randomly assigned to receive subcutaneous injections, three times/week for 30 days, of the following treatments: group 1 - 40 µg/kg of PTH (1-34) (n=15); group 2 - 2 µg/kg of PTH (1-34) (n=15) and; group 3 - only the vehicle required for hormone dissolution (n=15). Thirty days after surgery, the animals were sacrificed and specimens containing the implant and the surrounding bone were removed and processed for non-decalcified sections. The sections were evaluated according to the following histometric parameters: proportion of mineralized tissue (PMT) adjacent to the implant threads (500 µm band); bone filling within the limits of the threads (BF) and; bone-to-implant contact (BIC). For the cortical region, both hormone dosages (groups 1 and 2) promoted better results, for all parameters, when compared to control group (p<0.05). Similar results were observed for the BF parameter in the cancellous region (p=0.0394). Therefore, systemic administration of PTH (1-34) stimulates bone formation around titanium implants, even at low doses.

Sickle cell anemia, a genetic disease caused by a mutation in the beta-globin gene, can present oral manifestations such as delayed tooth eruption and hypomineralized enamel and dentin. The aim of the present study was to evaluate the prevalence and severity of developmental defects of enamel (DDE) and delayed tooth eruption in children with sickle cell anemia. The sample comprised 56 male and female children with sickle cell anemia aged 6 to 12 years and treated at the Hematology and Hemotherapy Center of Pernambuco, Brazil. The data were collected according to the WHO criteria for DDE and tooth eruption. The prevalence of DDE was 58.2% and increased with age, affecting 43.8% of children aged 6 to 8 years and 66.7% of those aged 10 to 12 years (p>0.05; Pearson's chi-square test). There was no significant association between DDE and sex; the most prevalent type of DDE was diffuse opacity (6.2%). Tooth eruption was delayed in 18 children (32.1%). The delay increased with age and was detected in 11.8% of children aged 6 to 8 years, in 20.0% of those aged 8 to 10 years and in 54.2% of those aged 10 to 12 years (p<0.05; Pearson's chi-square test). Delayed tooth eruption was higher in males (36.7%, p>0.05). The prevalence of DDE was high, increased with age and was similar between sexes, while delayed eruption was higher in males and showed a significant association with age.

This is a cross-sectional study that aimed to estimate maxillary sinus floor (MSF) pneumatization in single missing tooth of posterior maxilla, by using cone-beam computed tomography (CBCT). CBCT images were analyzed bilaterally and divided into 2 groups: edentulous site (EdS) - edentulous single region of upper second premolar, first or second molars; Tooth site (TS) - contralateral region homologous to the EdS region, with tooth present. Variables evaluated were: sinus height (SH), estimated sinus pneumatization (eSP: ∆ EdS - TS), healed ridge height (HR) and presence of localized sinus pneumatization (LSP) in molars teeth at TS. HR were categorized according to therapeutic option for posterior maxilla. 183 CBCT scans were included and it was observed that EdS presented a higher SH than the TS (p < 0.001) showing an eSP of 0.9 ± 2.93 mm. First molars presented the highest SH for both sides, although significant differences were detected when compared to second molars. First molars were mostly affected by LSP at TS (36 out of 43). Individuals with LSP at TS presented lower HR than the ones without LSP (p < 0.05). 54% of the cases presenting LSP obtained HR < 5 mm, which indicates sinus lift surgery. The present study showed that tooth loss in posterior maxilla favors sinus pneumatization and the identification of LSP at molar roots seems to indicate a greater necessity for sinus lift surgeries.

Background and Objectives: Semantic text annotation enables the association of semantic information (ontology concepts) to text expressions (terms), which are readable by software agents. In the scientific scenario, this is particularly useful because it reveals a lot of scientific discoveries that are hidden within academic articles. The Biomedical area has more than 300 ontologies, most of them composed of over 500 concepts. These ontologies can be used to annotate scientific papers and thus, facilitate data extraction. However, in the context of a scientific research, a simple keyword-based query using the interface of a digital scientific texts library can return more than a thousand hits. The analysis of such a large set of texts, annotated with such numerous and large ontologies, is not an easy task. Therefore, the main objective of this work is to provide a method that could facilitate this task., Methods: This work describes a method called Text and Ontology ETL (TOETL), to build an analytical view over such texts. First, a corpus of selected papers is semantically annotated using distinct ontologies. Then, the annotation data is extracted, organized and aggregated into the dimensional schema of a data mart., Results: Besides the TOETL method, this work illustrates its application through the development of the TaP DM (Target Prioritization data mart). This data mart has focus on the research of gene essentiality, a key concept to be considered when searching for genes showing potential as anti-infective drug targets., Conclusions: This work reveals that the proposed approach is a relevant tool to support decision making in the prioritization of new drug targets, being more efficient than the keyword-based traditional tools., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Alpinia purpurata is an ornamental crop known as a source of bioactive molecules. This is the first study to report isolation of a lectin (carbohydrate-binding protein) from A. purpurata inflorescences (ApuL). The immunomodulatory potential of ApuL was evaluated by investigating its effects on the production of cytokines and release of nitric oxide by human peripheral blood mononuclear cells (PBMCs). In addition, the differentiation and activation of lymphocytes treated with ApuL was evaluated by immunophenotyping assays. ApuL is an acidic and oligomeric protein with native molecular mass of 34kDa. The hemagglutinating activity (HA) of ApuL was inhibited by the glycoproteins fetuin and ovalbumin, was resistant to heating at 100°C and stimulated in the presence of calcium and magnesium ions. ApuL showed highest HA at pH 7.5 but failed to agglutinate erythrocytes at pH 8.0 and 9.0. ApuL induced the release of cytokines belonging to Th1 (IFN-γ, TNF-α, and IL-6) and Th17 (IL-17A) profiles as well as of nitric oxide, stimulating a pro-inflammatory environment. Moreover, ApuL also stimulated the production of IL-10, an anti-inflammatory cytokine with regulatory role. Incubation with lectin resulted in differentiation and activation of both T CD8 + and CD4 + subsets of lymphocytes, as evident from the expression of the CD28 costimulatory molecule. In conclusion, A. purpurata inflorescence is a source of an immunomodulatory lectin with potential immunoregulatory application, thereby adding biotechnological value to this ornamental crop., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Aims: To evaluate the use of the Portuguese version of the booklet "Your Pelvic Floor" as educational material, among Brazilian teenagers. The original version in English, directed to teenage audience, was developed by members of the Association for Continence Advice, in 2009., Methods: This quasi-experimental study was carried out at a non-governmental organization (NGO, São Paulo, Brazil) that develops continuing education programs for low-income girls aged eight to eighteen years old. A questionnaire with five questions was developed and validated by experts to measure the efficacy of the booklet "Your Pelvic Floor" on the learning of teenagers. After this, teenagers underwent three steps: pre-test, reading educational material and post-test, using the developed questionnaire to evaluate the knowledge about pelvic floor before and after reading the booklet., Results: Sixteen teenagers volunteered to participate. The number of correct answers to the questionnaire was significantly higher (P < 0.01) after reading the educational material, in all questions., Conclusions: The educational material "Your pelvic floor" was shown to be effective in teaching teenagers about the pelvic floor. Neurourol. Urodynam. 36:62-64, 2017. © 2015 Wiley Periodicals, Inc., (© 2015 Wiley Periodicals, Inc.)

A double blind randomized clinical trial of sufentanil as an adjunct in spinal anesthesia for cesarean section and, thereby, be able to reduce the dose of bupivacaine, a local anesthetic, with the same result of an anesthetic block with higher doses but with fewer perioperative side effects, such as hypotension., (Copyright © 2016 Sociedade Brasileira de Anestesiologia. Publicado por Elsevier Editora Ltda. All rights reserved.)

Chronic wounds are considered a worldwide epidemic and pain is the most frequently symptom referred by the patients who have this type of wound. The aim was to assess the prevalence of pain in chronic wounds and relate socio-demographic and clinical variables to pain. This is an analytical cross-sectional study, carried out in Basic Health Units (BHU), Family Health Strategy (ESF) and in a university hospital in two cities of Minas Gerais. 200 individuals who had chronic ulcers of different etiologies participated. Data collection took place between October 2012 and January 2013. The majority of participants (69%) reported they were in pain during the interview. Subjects with higher education had more pain. Patients with venous ulcers had less severe pain, and arterial ulcers were responsible for scoring the highest value of pain. It was found that most patients with chronic wounds feel pain; its intensity depends on the type of wound and this symptom should be recognized while caring for these patients., (Copyright © 2016 Tissue Viability Society. Published by Elsevier Ltd. All rights reserved.)

Scientific text annotation has become an important task for biomedical scientists. Nowadays, there is an increasing need for the development of intelligent systems to support new scientific findings. Public databases available on the Web provide useful data, but much more useful information is only accessible in scientific texts. Text annotation may help as it relies on the use of ontologies to maintain annotations based on a uniform vocabulary. However, it is difficult to use an ontology, especially those that cover a large domain. In addition, since scientific texts explore multiple domains, which are covered by distinct ontologies, it becomes even more difficult to deal with such task. Moreover, there are dozens of ontologies in the biomedical area, and they are usually big in terms of the number of concepts. It is in this context that ontology modularization can be useful. This work presents an approach to annotate scientific documents using modules of different ontologies, which are built according to a module extraction technique. The main idea is to analyze a set of single-ontology annotations on a text to find out the user interests. Based on these annotations a set of modules are extracted from a set of distinct ontologies, and are made available for the user, for complementary annotation. The reduced size and focus of the extracted modules tend to facilitate the annotation task. An experiment was conducted to evaluate this approach, with the participation of a bioinformatician specialist of the Laboratory of Peptides and Proteins of the IOC/Fiocruz, who was interested in discovering new drug targets aiming at the combat of tropical diseases., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Little is known about what possible effects on wood resources might be caused by non-timber forest products (NTFPs). Here, we assessed the patterns of fuelwood consumption related to an NTFP (Caryocar coriaceum) oil extraction and how this non-domestic activity can indirectly increase the use pressure on fuelwood species in a protected area, semiarid of Brazil. We conducted semi-structured interviews, in situ inventories, phytosociological surveys, and analyses of wood quality to identify the set of woody plants used in oil production. Householders use large volumes of dry wood and a set of woody species, which are highly exploited. Additionally, many preferred species have low fuel potential and suffer much use pressure. The best fuelwood species are underused, what requires management strategies to improve their potential as a source of energy. As a result, we suggest some conservation and management actions of fuelwood resources related to the use of NTFPs.

Feline sporotrichosis, which is caused by species of the Sporothrix schenckii complex, is endemic to Rio de Janeiro, Brazil. More than 4000 cases of the disease were diagnosed at Fundação Oswaldo Cruz, Brazil, between 1998 and 2012. Sporotrichosis in cats has been reported in several countries, but nowhere has an outbreak of animal sporotrichosis been as large as that seen in Brazil. The clinical manifestations of the disease range from an isolated skin lesion that can progress to multiple skin lesions and even fatal systemic involvement. Nodules and ulcers are the most common types of lesions, and respiratory signs and mucosa involvement are frequent. The definitive diagnosis depends on isolation of the etiologic agent in culture. Cytology, histopathology, and serology are useful tools for preliminary diagnosis. Severe pyogranulomatous inflammatory infiltrate, high fungal load, and extension of lesions to mucosa, cartilage, and bone in the nose of cats are indicative of an agent of high virulence in this endemic region. Itraconazole is the drug of choice, while, in refractory cases, amphotericin B or potassium iodide might be alternative treatments; however, recurrence after discharge may occur. Sporotrichosis persists as a neglected disease in Rio de Janeiro, and the treatment of cats remains a challenging and long-term endeavor., (© The Author 2014. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Purpose: To investigate the morphological aspects of brain and eyes in newborn rats whose mother underwent autogenously fecal peritonitis., Methods: Four pregnant rats that underwent fecal peritonitis, with a 10% fecal suspension in dose of 4 ml per kilogram received two antimicrobial treatments: 1. intraperitoneal moxifloxacin and dexamethazone; and 2. Intravenous meropenem. After head inspection, the brain consistencies and the eyes belonging to all offspring were analyzed., Results: The brains of newborn from rats that received 4 ml/kg of 10% suspension of feces showed, significantly smaller and less than the firm consistency of those in the control group. Congenital cataract was observed in 9 (34.6%). No cataract was observed in the 20 newborn rats from the mothers that received the combination of moxifloxacin and dexamethasone. Cataract could be observed in three (13.6%) offspring from mothers that received meropenem., Conclusions: Peritonitis can produce brain damage and congenital cataract in rats. The translation to humans is that intra abdominal infection in pregnant women may be associated with damage in brain and eye structures of their concepts. This can be averting using the adequate early therapeutically approach.

Purpose: To evaluate the treatment outcome of severe peritonitis in rats with increasing age., Methods: Thirty Wistar rats stratified in three groups: group I - six month-old; group II - 12 month-old; and group III - 18 month-old, underwent autogenously fecal peritonitis (6 ml/kg rat), and were treated with intravenous meropenem. The survival animals were followed-up for 45 days. The variables were expressed by their mean and standard error of the mean (SEM). p<0.05 was used for rejecting the null hypothesis. The study was approved by the Ethics Committee., Results: There was a significant increase in the mortality and morbidity in elderly rats. Of interest, even among young survival rats presenting with severe residual abscesses both in the abdomen and thorax cavities, they present an almost normal life., Conclusions: The treatment of severe autogenously fecal peritonitis with intravenous meropenem reached reasonable results in rats with six and twelve months of age, even considering residual abscesses on abdomen and thorax cavities. However, the great majority (80%) of elderly rats could not overcome the initial severe infectious challenge, proving that ageing is a very important risk factor for impairing immune response. Thus, sepsis remains a challenging situation, especially in elderly.

Background: The STINGRAY system has been conceived to ease the tasks of integrating, analyzing, annotating and presenting genomic and expression data from Sanger and Next Generation Sequencing (NGS) platforms., Findings: STINGRAY includes: (a) a complete and integrated workflow (more than 20 bioinformatics tools) ranging from functional annotation to phylogeny; (b) a MySQL database schema, suitable for data integration and user access control; and (c) a user-friendly graphical web-based interface that makes the system intuitive, facilitating the tasks of data analysis and annotation., Conclusion: STINGRAY showed to be an easy to use and complete system for analyzing sequencing data. While both Sanger and NGS platforms are supported, the system could be faster using Sanger data, since the large NGS datasets could potentially slow down the MySQL database usage. STINGRAY is available at http://stingray.biowebdb.org and the open source code at http://sourceforge.net/projects/stingray-biowebdb/.

Parkia pendula seed lectin was used to treat cutaneous wounds of normal and immunocompromised mice, inducing cicatrization. Methotrexate (0.8 mg/kg/week) was used as immunosuppressive drug. Wounds were produced in the dorsal region (1 cm(2)) of female albino Swiss mice (Mus musculus), health and immunocompromised. Wounds were daily topically treated with 100 μL of the following solutions: (1) control (NaCl 0.15 M), (2) control Im (0.15 M NaCl), (3) P. pendula seed lectin (100 μg/mL), and (4) P. pendula seed lectin Im (100 μg/mL). Clinical evaluation was performed during 12 days. Biopsies for histopathology analysis and microbiological examinations were carried out in the second, seventh, and 12th days. The presence of edema and hyperemia was observed in all groups during inflammatory period. The first crust was detected from the second day, only in the groups treated with P. pendula seed lectin. Microbiological analysis of wounds from day 0 to day 2 did not show bacterium at P. pendula seed lectin group; however, Staphylococcus sp. was detected every day in the other groups. The lectin markedly induced a total wound closing at P. pendula seed lectin and P. pendula seed lectin Im groups on 11th day of evolution. The present study suggests that P. pendula seed lectin is a biomaterial potential to show pharmacological effect in the repair process of cutaneous wounds.

Purpose: To evaluate the treatment outcome of severe peritonitis in rats submitted to permanent bilateral carotid occlusion (PBCO)., Methods: Sixteen Wistar rats (mean age of 8.5 months) with PBCO underwent autogenously fecal peritonitis, and were treated with moxifloxacin combined with dexamethasone, and followed-up for 45 days. Ten rats (mean age five months) without PBCO were used as a control group. The variables were expressed by their mean and standard error of the mean (SEM). p<0.05 was used for rejecting the null hypothesis. The study was approved by the Ethics Committee., Results: There was a significant increase (p=0.0002) in the mortality and morbidity in older rats that underwent PBCO (study group). However, even among the survival rats presenting with severe residual abscesses both in the abdomen and thorax cavities, they present an almost normal life., Conclusions: The treatment of severe autogenously fecal peritonitis with intraperitoneal moxifloxacin combined with dexamethasone was very effective in young rats without permanent bilateral carotid occlusion. The treatment reached reasonable results in older rats with PBCO, even considering residual abscesses on abdomen and thorax. Older age was the greater risk factor for the outcome of the treatment of severe peritonitis. Sepsis remains a challenging situation, especially in elderly.

Purpose: To develop an alcoholic extract of the inner bark of the Schinus terebinthifolius raddi and to test its impact on autogenously fecal peritonitis in Wistar rats., Methods: The inner bark of the Schinus terebinthifolius raddi was kept for seven days in 70% ethanol alcohol. The total elimination of the solvent was performed in a rotary evaporator under reduced pressure at 55-60°C. Four milliliter of this extract was injected, after 24 h, into the abdominal cavity of six out of eight survival rats that underwent autogenously fecal peritonitis with five milliliter of 10% filtered fecal suspension. They were clinically followed up for 45 days when they were euthanized. The necropsy findings (inventory) of the abdominal and thorax cavities were inspected and the main findings were recorded and photographed. The investigation was approved by the Ethics Committee., Results: Two out of six survival rats that were critically ill after 24 h died within the 12 h after the extract injection into the abdominal cavity. Four rats that were also critically ill recovered and gradually became healthy, eating well, regaining weight and moving normally in the cage. At 45 days post severe peritonitis the necropsy findings revealed few signs of residual infection on the abdominal and thorax cavities. There were no bowel adhesions., Conclusion: The impact of alcoholic extract of the inner bark of the Schinus terebinthifolius raddi was considered very positive and promising as natural local antiseptic against very severe peritonitis in Wistar rats.

Purpose: To evaluate the effects of chronic cerebral hypoxia on memory of rats submitted to bilateral common carotid artery ligation (BCCL)., Methods: Every each week, for 16 weeks, 31 rats were tested for memory using a water and land mazes and compared with 30 normal rats (control group A). The variables were expressed by their mean and standard error of the mean (SEM). p<0.05 was used for rejecting the null hypothesis. The study was approved by the Ethics Committee for animal investigation., Results: There was a significant increase in the latency time, in the survival water and land mazes, after four weeks (study group B) follow-up. However, without any medication or therapeutically induced measures, after 16 weeks (study group C) follow-up the latency mean times tend to be similar to control group (A) in the neurocognitive tests., Conclusions: Neurocognitive deficits after 16 weeks post-operative follow-up of rats that underwent bilateral common carotid artery ligation is a natural adaptive phenomenon. Thus, is not realistic to allow translational information from this animal model for therapeutically approaches aiming at to prevent, or to improve brain damage in human beings suffering from chronic deprivation of adequate blood supply.

Background: The extract from Moringa oleifera seeds is used worldwide, especially in rural areas of developing countries, to treat drinking water. M. oleifera seeds contain the lectins cmol and WSMoL, which are carbohydrate-binding proteins that are able to reduce water turbidity because of their coagulant activity. Studies investigating the ability of natural products to damage normal cells are essential for the safe use of these substances. This study evaluated the cytotoxic and anti-inflammatory properties of the aqueous seed extract, the extract used by population to treat water (named diluted seed extract in this work), and the isolated lectins cmol and WSMoL., Methodology/principal Findings: The data showed that the aqueous seed extract and cmol were potentially cytotoxic to human peripheral blood mononuclear cells, while WSMoL and diluted seed extract were not cytotoxic. The M. oleifera aqueous seed extract and the lectins cmol and WSMoL were weakly/moderately cytotoxic to the NCI-H292, HT-29 and HEp-2 cancer cell lines and were not hemolytic to murine erythrocytes. Evaluation of acute toxicity in mice revealed that the aqueous seed extract (2.000 mg/kg) did not cause systemic toxicity. The aqueous seed extract, cmol and WSMoL (6.25 µg/mL) and diluted seed extract at 50 µg/mL exhibited anti-inflammatory activity on lipopolyssaccharide-stimulated murine macrophages by regulating the production of nitric oxide, TNF-α and IL-1β. The aqueous seed extract reduced leukocyte migration in a mouse model of carrageenan-induced pleurisy; the myeloperoxidase activity and nitric oxide, TNF-α and IL-1β levels were similarly reduced. Histological analysis of the lungs showed that the extract reduced the number of leukocytes., Conclusion/significance: This study shows that the extract prepared according to folk use and WSMoL may be non-toxic to mammalian cells; however, the aqueous seed extract and cmol may be cytotoxic to immune cells which may explain the immunosuppressive potential of the extract.

Purpose: To conduct a systematic literature review focusing on the use of virtual reality (VR) for the improvement of gait in post-stroke patients., Methods: We performed a search of Randomized-controlled trials published from 1966 to 2011 in the databases: Medline, Lilacs, CINAHL, Cochrane and SciELO. Keywords used in the selection were: Virtual reality in combination with (AND) "Nervous System Diseases", (OR) "Motor Skill Disorders" (OR) "neurologic impairments" (OR) "motor function" (OR) function* (OR) locomotion (OR) ambulation (OR) gait (OR) "motor activity" (OR) Stroke. Selected articles were evaluated using the individual's components of methodological quality assessment and analysis of outcomes of each study was based on the domains of the International Classification of Functioning, Disability and Health (ICF)., Results: A total of 6520 references were found, however, based on the inclusion and exclusion criteria only four studies were considered and analyzed. These articles demonstrated that the use of VR promotes changes in gait parameters, despite the diversity of protocols, participants' characteristics, as well as the number of participants included in each study., Conclusions: The research studies analyses suggest that VR is a promising method to improve the gait of patients with stroke. Nevertheless, some questions still need to be answered. Some aspects should be investigated to confirm the true benefits and application of VR in this population., Implications for Rehabilitation: Stroke is the second cause of death and the leading cause of disability worldwide. The loss or impairment of ambulation is one of the most devasting sequelae of stroke. Restoration of gait can be considered the main goal of rehabilitation after stroke. Conventional interventions tend to be tedious, providing few opportunities to increase the difficulty level of the proposed tasks and do not encourage adaptive postural reactions. There is evidence to support the use of virtual reality for the promotion of walking in people with sequelae of stroke. Virtual reality is a feature that has been used in clinical practice, however, the details on how to use this instrument must be set according to the therapeutic goals.

Purpose: To investigate morbidity, mortality and microbiological response to fecal peritonitis induced in Wistar rats with permanent bilateral carotid ligation (PBCL)., Methods: Fecal peritonitis was induced in 30 rats, with 10 animals in each group: Group1 - normal young animals; Group2 - normal mature animals; and Group3 - rats with PBCL after four months postoperative follow-up. Peritonitis was induced with 10% stool suspension. Morbidity and mortality were evaluated. The survival animals after seven days were euthanized for tests. For microbiological studies blood were collected from the carotids and right ventricle; and fragments of lung and peritoneum., Results: The morbidity and mortality of young animals were significantly lower than in mature animals with and without PBCL. There was no difference in morbidity and mortality among mature rats with and without PBCL. The diversity of microorganisms producing septicemia was similar to native micro biota of the large bowel., Conclusions: The immune response was more efficient in young animals, represented by significant less morbidity and no natural mortality. PBLC did not affect morbidity and mortality in mature rats. The immune response to fecal peritonitis has age as an independent predictor.

Purpose: To evaluate the effect of cerebral hypoxia-ischemia on memory and learning survival of rats submitted to permanent bilateral carotid ligation (PBCL)., Methods: Twenty-four survivors of PBCL were evaluated after 30 days with regard to memory and learning using a water survival maze. Twenty-three healthy rats were used as control group. The results were expressed by their means and standard error of the mean (SEM). p<0.05 was used for rejecting the null hypothesis. The study was approved by the Ethics Committee for animal investigation., Results: The mortality rate for the surgery was 44.4%. The latency time to find the survival platform was higher in rats that underwent PBCL (Normal: 10.24 ± 1.85s -, Study: 25.30 ± 4.69s - Mann - Whitney p=0.0388). Additionally, the type of swimming and the spatial stability of the studied rats on the survival platform were compromised in these animals., Conclusion: The permanent bilateral carotid ligation induces change in the learning and survival memory.

This work reports the isolation of a serum lectin from cobia fish (Rachycentron canadum) named RcaL. Immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production were also performed. RcaL was obtained through precipitation with ammonium sulphate and affinity chromatography on a Concanavalin A-Sepharose 4B column. The ammonium sulphate fraction F3 showed the highest specific hemagglutinating activity and was applied to affinity chromatography. The lectin was eluted with methyl-α-D-mannopyranoside. RcaL showed highest affinity for methyl-α-D-mannopyranoside and D-mannose; eluted fractions of RcaL agglutinated rabbit erythrocytes (titre, 128(-1)) retained 66 % of chromatographed lectin activity, and the obtained purification factor was 1.14. Under reducing conditions, a polypeptide band of 19.2 kDa was revealed in sodium dodecyl sulphate polyacrylamide gel electrophoresis (PAGE). PAGE confirmed RcaL as an acidic protein revealed in a single band. Cytotoxic and immunomodulatory assays with RcaL in mice splenocyte cultures showed that the lectin was not cytotoxic and induced higher interferon gamma and nitric oxide production in splenocyte cultures. Purified RcaL induced preferential Th1 response, suggesting that it acts as an immunomodulatory compound.

Objective: To analyze the impact of moderate physical exercise on the total and differential leukocyte counts and red blood cell count of 36 sixty-day-old adult male Wistar rats subjected to early malnourishment., Methods: The rats were divided in nourished (N - casein 17%) and malnourished groups (M - casein 8%) and thesegroups were then subdivided in trained (T) untrained (U) creating four groups NT, NU, MT and MU. The NT and MTgroups were submitted to moderate physical exercise using a treadmill (60 min/day, 5 days/week for 8 weeks). Onthe 1st day, before the training started T0 and 24 hours after the last training day of the week (T1 until T8), a 1 mLaliquot of blood was collected from the animals' tails for analysis. The total leukocyte count was evaluated in a cellcounter with an electronic microscope. The cyanmethemoglobin technique was used to measure the hemoglobin level. The hematocrit values were determined as a percentage using the micro-hematocrit technique with a microcapillaryreader and a cell counter was used to determine the red blood cell count. The t-test was used for statistical analysis and a p-value < 0.05 was considered significant. Data are expressed as means ± standard deviation., Results: There was a significant difference in the total leukocyte count between the NT (9.1 ± 0.1) and MT groups (8.0 ± 0.1) from T1 and in neutrophils between the NT (22.1 ± 0.6) and MT groups (24.6 ± 1.8) from T7 (p < 0.05). There was no statistical significance in the hemoglobin, hematocrit and red blood cell count from T1., Conclusions: According to the results of this study, moderate physical exercise seems to have induced physiologic adaptation in adult rats from T1.

The aim of this work was to purify and partially characterize a mannose recognition lectin from Nile tilapia (Oreochromis niloticus) serum, named OniL. OniL was isolated through precipitation with ammonium sulfate and affinity chromatography (Concanavalin A-Sepharose 4B). In addition, we evaluated carbohydrate specificity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profiles, and in vitro immunomodulatory activity on mice splenocyte experimental cultures through cytotoxic assays and cytokine production. The ammonium sulfate fraction F2 showed the highest specific hemagglutinating activity (331) and was applied to affinity matrix. Adsorbed proteins (OniL) were eluted with methyl-α-D: -mannopyranoside. OniL, a 17-kDa protein by SDS-PAGE constituted by subunits of 11 and 6.6 kDa, showed highest affinity for methyl-α-D: -mannopyranoside and D: -mannose. Immunological assays, in vitro, showed that OniL did not show cytotoxicity against splenocytes, induced higher IFN-γ production and lower IL-10 as well as nitrite release. In conclusion, OniL lectin was successfully purified and showed a preferential Th1 response in mice splenocytes.

Management of male infertility largely depends on our understanding of cellular and molecular aspects of spermatogenesis as well as sperm function. Apart from standardized and comprehensive semen analysis, prognostic markers estimating the fertilizing capacity of either ejaculated spermatozoa or testicular spermatids are required. While there is general agreement that correct replacement of DNA-binding histones by protamines represents a prerequisite for achieving competent spermatozoa, especially in intracytoplasmic sperm injection (ICSI) where natural selection mechanisms are bypassed, the function of a variety of transcripts within the spermatozoa's cytoplasm and of remaining highly acetylated histones is still a matter of debate. Hence, this review brings the up-to-date research on mammalian spermatozoal chromatin composition into focus, which is discussed in conjunction with the paternal role on epigenetic reprogramming of the zygote following fertilization. As paternal transcripts have been demonstrated to be transmitted to the oocyte, it is now accepted that they represent more than solely remnants of previous transcriptional activity. Acetylation of histones, normaly a characteristic of transcriptional activity, was for a long time a miracle, as spermatozoa are transcriptionally inactive cells, but is now suggested to represent epigenetic marks that are transmitted to the oocyte and play an important role in the regulation of early gene expression in the developing embryo., (© 2010 The Authors. International Journal of Andrology © 2011 European Academy of Andrology.)

Beta-actin (ACTB), glyceraldehyde-3-phosphate-dehydrogenase (GAPD), Heat Shock Protein 1, beta (HSPCB) and Adenosine Triphosphate subunit 5 beta (ATP5B) with distinct functional characteristics and expression patterns were investigated as suitable references for gene expression studies. We determined the expression stability of the four reference genes in ejaculates, cryopreserved as well as fixed and paraffin-embedded testicular tissue (from fertile and subfertile men) applying real-time qRT-PCR and statistical analysis. The mean gene expressions (mean Ct value) were compared for each gene between the fertile and subfertile donors by using the Wilcoxon-Mann-Whitney test. We did not observe significant statistical differences between variability of genes. To detect random effects, we used the two-way analysis of variance with a hierarchical model. The results show no significant statistical differences between proband and repetition within the probands. Taken together, we concluded that ACTB, GAPD, HSPCB and ATP5B have a variable expression within these samples, but this variability is not statistically significant. This finding demonstrated that all these genes could be appropriated for further studies on gene expression in ejaculate and testis tissue. Therefore, the selection of the suitable reference genes is highly specific for a particular experimental model and validation for each situation, on an individual basis, is a crucial requirement., (© 2011 Blackwell Verlag GmbH.)

Human sperm contain similar amounts of protamine-1 (P1) and protamine-2 (P2). Although an aberrant protamine ratio have been observed in subfertile men, functional evidence is provided by protamine knockout mice exhibiting male infertility. As sperm DNA integrity is known to be linked with DNA fragmentation and apoptosis, we investigated whether the DNA fragmentation factor 40 (DFF40) ratio or caspase (Casp4, Casp6) and tumor necrosis factor superfamily member 10 (TNFSF10) ratio together with the P1/P2 ratio represents a reliable biomarker to discriminate between fertile and subfertile men. Real-time quantitative RT-PCR was used for amplification of P1, P2 and DFF40 in 49 testicular biopsies. Casp4, Casp6 and TNFSF10 have been selected from a PCR apoptosis array and were further investigated in another group of testicular biopsies (22 subfertile men versus 11 potentially fertile men). Using Spearman's rank correlation coefficient analysis, we did not find a correlation between DFF40 and P1, P2, P1/P2, score, fertilization rate and age. In addition, logistic regression analysis demonstrated no statistically significant effect of the analyzed variables on pregnancy. A two-way analysis of variance with repeated measures of relative expression of Casp4, Casp6 and TNFSF10 versus P1 or P2 in potentially fertile men and subfertile patients demonstrated statistically significant differences between both groups, all tested gene combinations and the interaction between two genes and both groups in all cases analyzed. Furthermore, significant differences in the expression of Casp4 and TNFSF10 between the groups of potentially fertile and subfertile men could be demonstrated. In addition, the means of differences of selected gene combinations revealed that the protamine to apoptotic gene ratio is statistically different between both groups. Our data suggest that Casp4, Casp 6 and TNFSF10 are differentially expressed in potentially fertile and subfertile men and represent useful biomarkers for predicting male fertility in combination with P1 and P2.

Spermatozoa are transcriptionally inactive cells, but contain acetylated histones, normally a characteristic of transcriptionally active cells. Acetylgroups are thought to represent epigenetic marks that are transmitted to the oocyte and are involved in starting gene expression in the zygote and in regulating gene expression during early embryogenesis. We performed reverse transcription polymerase chain reaction (RT-PCR) in the common marmoset monkey (Callithrix jacchus) and in bovine spermatozoa, oocytes, zygotes, two- and four-cell embryos to evaluate the presence of specific transcripts known to play a role during fertilisation and early embryo development, namely protamine-1 (PRM1), protamine-2 (PRM2), histone H1 (H1), histone H3 (H3), histone H4 (H4), cAMP-responsive element modulator (CREM), DNA methyltransferase-1 (DNMT1), TATA box-binding protein (TBP). All transcripts tested were present in spermatozoa of the common marmoset, while bull spermatozoa lack PRM2. Marmoset oocytes exhibited transcripts for H1, H3, H4 and TBP, whereas bovine oocytes revealed H1, H3, H4, CREM, DNMT and TBP mRNAs. In zygotes, we amplified H1, H4, TBP (marmoset) and PRM1, H1, H3, H4, CREM, DNMT1 and TBP (bovine). Two-cell embryos showed PCR products for H1, H3 and TBP in the marmoset. In the bovine, all transcripts could be observed except PRM2. In four-cell embryos, PCR signals were obtained for PRM1, H1, H3, H4 and TBP in the marmoset. In the bovine, all transcripts were detected except PRM2. Our data suggest that, in both C. jacchus and Bos taurus, PRM1 transcripts are delivered by the spermatozoon to the oocyte., (© 2011 Blackwell Verlag GmbH.)

There is no adequate animal model to mimic the difficult clinical situation of infected non-union of the tibia after intramedullary stabilization. The purpose was to establish an animal model of implant-related infected non-unions of the tibia in rats. Furthermore, it was evaluated if detection of bacteria by fluorescent in situ hybridisation (FISH) technique is possible in bone infection. 17 rats were used in which osteotomy of the midshaft tibia was performed and stabilized with an intramedullary device. Two groups were tested: group 1: contamination of the osteotomy site with 10(4) colony forming units (CFUs) of Staphylococcus aureus (11 animals), group 2: no bacterial contamination (6 animals). The animals were sacrificed after 42 days and bone healing and infection were assessed clinically, by X-ray, micro-CT, and microbiological methods including FISH technique using EUB and STAPHY probes. Histology and scanning electron microscopy (SEM) for biofilm formation were performed. All animals of the control group showed uneventful bone healing after 6 weeks without any signs of local infections. 10 of 11 (90.9%) animals of group 1 with bacterial contamination exhibited infected non-union formation with positive clinical, radiological and microbiological infection signs of the tibia but without any systemic infection signs. FISH technique was able to identify bacteria in the infected bone. All intramedullary implants from the infected animals showed positive biofilm formation in SEM. This work presents the first animal model for the induction of intramedullary device-related infected non-union in the tibia and detection of bacteria by FISH technique in infected bone., (Copyright © 2011 Elsevier Inc. All rights reserved.)