Study of the immunologic response of marine-derived collagen and gelatin extracts for tissue engineering applications.

The host immunologic response to a specific material is a critical aspect when considering it for clinical implementation. Collagen and gelatin extracted from marine sources have been proposed as biomaterials for tissue engineering applications, but there is a lack of information in the literature about their immunogenicity. In this work, we evaluated the immune response to collagen and/or gelatin from blue shark and codfish, previously extracted and characterized. After endotoxin evaluation, bone marrow-derived macrophages were exposed to the materials and a panel of pro- and anti-inflammatory cytokines were evaluated both for protein quantification and gene expression. Then, the impact of those materials in the host was evaluated through peritoneal injection in C57BL/6 mice. The results suggested shark collagen as the less immunogenic material, inducing low expression of pro-inflammatory cytokines as well as inducible nitric oxide synthase (encoded by Nos2) and high expression of Arginase 1 (encoded by Arg1). Although shark gelatin appeared to be the material with higher pro-inflammatory expression, it also presents a high expression of IL-10 (anti-inflammatory cytokine) and Arginase (both markers for M2-like macrophages). When injected in the peritoneal cavity of mice, our materials demonstrated a transient recruitment of neutrophil, being almost non-existent after 24 hours of injection. Based on these findings, the studied collagenous materials can be considered interesting biomaterial candidates for regenerative medicine as they may induce an activation of the M2-like macrophage population, which is involved in suppressing the inflammatory processes promoting tissue remodeling. STATEMENT OF SIGNIFICANCE: Marine-origin biomaterials are emerging in the biomedical arena, namely the ones based in marine-derived collagen/gelatin proposed as cell templates for tissue regeneration. Nevertheless, although the major cause of implant rejection in clinical practice is the host's negative immune response, there is a lack of information in the literature about the immunological impact of these marine collagenous materials. This work aims to contribute with knowledge about the immunologic response to collagen/gelatin extracted from blue shark and codfish skins. The results demonstrated that despite some differences observed, all the materials can induce a macrophage phenotype related with anti-inflammation resolution and then act as immuno-modulators and anti-inflammatory inducible materials.

Allergic sensitization to Storage Dust Mites: a prospective study of patients with respiratory allergy

Summary: Objectives. To describe the prevalence of allergic sensitization to Storage Dust Mites (SDM), access whether the place of living and occupational exposure were determinants for SDM sensitization and study association between Lepidoglyphus destructor and other SDM sensitization. Methods. Prospective analysis of patients evaluated for suspected allergic rhinitis and/or asthma that performed Skin Prick Tests (SPT) to SDM between January and December 2018 in our Department. Results. Two hundred consecutive patients were evaluated for rhinitis and/or asthma in our outpatient consultation: 123 (61.5%) presented positivity for at least one SDM, 68.3% were female and the mean age was 33.1 ± 12.12. Lepidoglyphus destructor (69.9%) was the most prevalent, followed by Tyrophagus putrescentiae (50.4%), Blomia tropicalis and Glycyphagus domesticus (48.8%) and Acarus siro (24.4%). Living in a rural place was not associated with a higher prevalence of sensitization to SDM, except for Acarus siro (p = 0.032), and working in a place with storage areas was not associated with sensitization to any of SDM. Sensitization to Lepidoglyphus destructor was associated with sensitization to Blomia tropicalis, Glycyphagus domesticus and Tyrophagus putrescentiae (p less than 0.005), but not with Acarus siro.Conclusions. Our study suggests that our population, independently of their occupational exposure and place of residency, are sensitized to SDM and that evaluation of sensitization to SDM should be considered as standard practice.

Achados clínicos, histopatológicos e moleculares da mielopatia degenerativa canina: relato de caso / Clinical, histopathological and molecular findings of canine degenerative myelopathy: case report

Objetivou-se descrever os achados clínicos, histopatológicos e moleculares associados à MDC em um cão da raça Pastor-Suiço. O cão possuía uma paraparesia progressiva em membros pélvicos e foi submetido a avaliações clínicas, pelas quais se obteve, entre outros diferenciais, o diagnóstico presuntivo de MDC. Com a evolução dos sinais, o tutor optou pela eutanásia. Os achados histopatológicos da medula espinhal foram compatíveis com uma degeneração segmentar axonal e mielínica. O diagnóstico molecular foi realizado por meio da extração do DNA obtido por swab oral. Uma PCR foi otimizada utilizando-se primers descritos em literatura para amplificar a região do gene SOD1. A amostra foi, então, submetida a sequenciamento unidirecional, que revelou que o animal em questão era homozigoto para o alelo A para a mutação c.118G>A no éxon 2 do gene SOD1. O diagnóstico clínico presuntivo da MDC no presente caso foi esclarecido por meio dos achados histopatológicos, associados aos achados clínicos, e da sua caracterização molecular. Ressalta-se a contribuição deste relato, que traz aspectos clínicos, histopatológicos e moleculares associados à MDC na raça Pastor-Suíço, para a qual, até o presente momento, na literatura consultada, não há relato dessa enfermidade.(AU)

The objective of this study was to describe the clinical, histopathological and molecular findings associated with MDC in a Swiss Shepherd dog. The dog had a progressive paraparesis in pelvic limbs and was submitted to clinical evaluations where, among other differentials, the presumptive diagnosis of MDC was obtained. With the progression of the nervous deficits tutor opted for euthanasia. The histopathological findings of the spinal cord were compatible with axonal and myelinic segmental degeneration. Molecular diagnosis was performed by extracting the DNA obtained by oral swab. PCR was optimized using primers described in the literature to amplify the SOD1 gene region. The sample was then subjected to one-way sequencing which revealed that the animal in question was homozygous for the A allele for the c.118G>A mutation in exon 2 of the SOD1 gene. The presumptive diagnosis of MDC in the present case was clarified by histopathological findings, as well as by its molecular characterization. The contribution of this report brings clinical, histopathological and molecular aspects associated with canine degenerative myelopathy in the Swiss Shepherd breed, that until this moment, in the literature consulted, there is no report of this disease in the breed mentioned.(AU)

Prevalence of apical periodontitis and its association with previous root canal treatment, root canal filling length and type of coronal restoration - a cross-sectional study.

AIM: To analyse the prevalence of periapical lesions and their association with previous root canal treatment, root canal filling length and type of coronal restoration using in vivo cone-beam computed tomographic (CBCT) assessment. METHODOLOGY: A global sample of 20 836 teeth, with a combined total of 27 046 roots, from 1160 patients, was analysed via CBCT assessment in eight health centres. Each tooth was evaluated by one out of five examiners after having performed a defined calibration procedure on the basis of 319 teeth. Intra- and inter-rater reliability tests were performed. Each tooth was classified according the tooth number, presence/absence of periapical lesions, presence/absence of previous root canal treatment, length of root canal filling (short, good or overfilling) and type of coronal restoration. The z-test for proportions was used to analyse differences between tooth subgroups, and an odds ratio was determined in order to analyse the association between treatment status and periapical lesions. A P < 0.05 was considered significant. RESULTS: At a tooth level, the overall prevalence of periapical lesions in the sample was 10.4%. Maxillary teeth were associated with a significantly larger percentage of lesions (13.1%), whilst maxillary first molars had the greater proportion of lesions (21.2%). The prevalence of periapical lesions was significantly larger in root filled teeth (55.5%), short root canal fillings (72.7%) and in teeth restored with crowns (46.1%). At a root level, the mesiobuccal roots of both maxillary first molars had a tendency for a larger percentage of periapical lesions. CONCLUSION: History of root canal treatment, root canal filling length and type of coronal restoration influenced the presence of periapical lesions. Molars were more commonly associated with periapical lesions on root filled teeth, particularly those with short root fillings and those with crowns.

Heparin Resistance During Surgical Resection of Inferior Vena Cava and Right Atrial Tumor Thrombus: A Case Report.

INTRODUCTION: Heparin resistance is the failure of unusually high doses of heparin to achieve a target activated clotting time (ACT). We present the case of a 47-year-old female patient, ASA 2, who was diagnosed with a neuroendocrine retroperitoneal tumor with thrombus in the left renal vein, inferior vena cava and right atrium. General and cardiothoracic surgeons collaborated to remove the tumor under cardiopulmonary bypass. Heparin resistance was considered and treated with 1000 UI of antithrombin III concentrate. The authors review the mechanisms and management of this entity.

The infertility trap: how defeat and entrapment affect depressive symptoms.

STUDY QUESTION: Does the perception of failure without a solution or way forward of infertile couples have a mediator role between the importance couples attribute to parenthood and depressive symptoms? SUMMARY ANSWER: The perception of failure without a solution or way forward, assessed by feelings of entrapment and defeat, mediates the effect of the importance of parenthood on depressive symptoms of infertile men and women. WHAT IS KNOWN ALREADY: Research has documented that the heightened importance of parenthood affects infertile couples' adjustment to infertility and medical treatments. However, it remains unclear which psychological mechanisms and perceptions may underlie the association between having parenthood as a nuclear aspect of life and presenting depressive symptoms related to difficulties in accomplishing that important life goal. Although these links have been scantly addressed in infertility, previous studies have pointed to the role that perceptions of defeat and entrapment have in several psychopathological conditions. STUDY DESIGN, SIZE, DURATION: The study was cross-sectional. Couples pursuing medical treatment for their fertility problems were invited to participate by their doctors in several public and private clinics. Data collection took place between July 2009 and 2011. PARTICIPANTS/MATERIALS, SETTING, METHODS: One hundred forty-seven infertile couples consented to participate in the study. Both couple members (147 women and 147 men) completed a set of self-report instruments for the assessment of depressive symptoms, perceptions of defeat and entrapment, importance of parenthood and rejection of a childfree lifestyle. Analyses were conducted through Structural Equation Modeling and followed a dyadic analysis strategy, allowing for controlling the interdependence of the data. MAIN RESULTS AND THE ROLE OF CHANCE: The hypothesized tested model showed a very good fit to the data [(χ(2) = 68.45, P = 0.014, comparative fit index = 0.98, standardized root-mean-square residual = 0.06 and root mean square error of approximation = 0.06] and explained 67 and 58% of the variability in depressive symptoms in women and men, respectively. Results revealed that the importance of parenthood does not have a direct effect on depressive symptoms of infertile men and women, but an indirect effect, by affecting the perception of having failed and not being able to solve it or move forward [women: estimate for indirect effect: 0.38 (bias corrected (BC) 95% confidence interval (CI) = 0.25; 0.56; P < 0.001); men: estimate for indirect effect: 0.23 (BC 95% CI = 0.06; 0.40; P = 0.013)]. LIMITATIONS, REASONS FOR CAUTION: The study was cross-sectional, which does not allow for the establishment of causality. Another limitation is the heterogeneity of the sample, as participants were recruited at various stages of their fertility care. In addition, due to the specific nature of the variables, further studies are needed to establish exactly how the relationship between defeat and entrapment and depression operates, as the mechanism may be bidirectional. WIDER IMPLICATIONS OF THE FINDINGS: This study emphasizes the role of perceptions of defeat and entrapment on the psychological adjustment to infertility and assisted reproduction. These emotional processes should be taken into consideration and targeted in psychological interventions of couples undergoing medical treatments for infertility. In fact, although parenthood may be perceived as a core purpose for many couples dealing with difficulties in conceiving, it is only when these difficulties are experienced as failures without a resolution and as inescapable, that couples are prone to develop depressive symptoms. STUDY FUNDING/COMPETING INTERESTS: This research has been supported by the first author Ph.D. Grant (SFRH/BD/68392/2010), sponsored by the Portuguese Foundation for Science and Technology (FCT). The authors have no conflict of interests. TRIAL REGISTRATION NUMBER: N/A.

Origami rules for the construction of localized eigenstates of the Hubbard model in decorated lattices.

We present a method of construction of exact localized many-body eigenstates of the Hubbard model in decorated lattices, both for U = 0 and U → ∞. These states are localized in what concerns both hole and particle movement. The starting point of the method is the construction of a plaquette or a set of plaquettes with a higher symmetry than that of the whole lattice. Using a simple set of rules, the tight-binding localized state in such a plaquette can be divided, folded and unfolded to new plaquette geometries. This set of rules is also valid for the construction of a localized state for one hole in the U → ∞ limit of the same plaquette, assuming a spin configuration which is a uniform linear combination of all possible permutations of the set of spins in the plaquette.

A novel configuration for an anaerobic submerged membrane bioreactor (AnSMBR). Long-term treatment of municipal wastewater under psychrophilic conditions.

A novel design for a pilot scale anaerobic submerged membrane bioreactor (AnSMBR) equipped with an ultrafiltration unit, treating municipal wastewater at 18 ± 2°C, and inoculated with a mesophilic inoculum without acclimation, was implemented and evaluated over 3 years of stable operation. The AnSMBR operated with a volumetric loading rate between 1.6 to 2.0 kg COD/m(3)UASBd, 12.8 to 14.2h hydraulic retention time, and reached a tCOD removal efficiency of around 90%. Biosolid production was between 0.05 and 0.083 g VS/g CODremoved. Dissolved methane oversaturation in the effluent was observed, reaching average values of 19.1 ± 0.84 mg CH4/L. The permeate flow rate ranged from 10 to 14L/m(2)h with trans-membrane pressure (TMP) values of 400-550 mbar, using cycles of 30s backwash, 7.5 min filtration, and continuous biogas sparging (9-16 m/h). During the three years of continuous operation, the membrane was not physically or chemically cleaned.

Long-term operation of a pilot scale anaerobic membrane bioreactor (AnMBR) for the treatment of municipal wastewater under psychrophilic conditions.

The performance of a pilot scale anaerobic membrane bioreactor (AnMBR), comprising an upflow anaerobic sludge blanket (UASB) reactor coupled to an external ultrafiltration membrane treating municipal wastewater at 18±2°C, was evaluated over three years of stable operation. The reactor was inoculated with a mesophilic inoculum without acclimation. The AnMBR supported a tCOD removal efficiency of 87±1% at hydraulic retention time (HRT) of 7h, operating at a volumetric loading rate (VLR) of between 2 and 2.5kgtCOD/m(3)d, reaching effluent tCOD concentrations of 100-120mg/L and BOD5 concentrations of 35-50mgO2/L. Specific methane yield varied from 0.18 to 0.23Nm(3)CH4/kgCODremoved depending on the recirculation between the membrane module and the UASB reactor. The permeate flow rate, using cycles of 15s backwash, 7.5min filtration, and continuous biogas sparging (40-60m/h), ranged from 10 to 14Lm(2)/h with trans-membrane pressure (TMP) values of 400-550mbar.

N-cinnamoylated aminoquinolines as promising antileishmanial agents.

A series of cinnamic acid conjugates of primaquine and chloroquine were evaluated for their in vitro antileishmanial activities. Although primaquine derivatives had modest activity, chloroquine conjugates exhibited potent activity against both promastigotes (50% inhibitory concentration [IC50] = 2.6 to 21.8 µM) and intramacrophagic amastigotes (IC50 = 1.2 to 9.3 µM) of Leishmania infantum. Both the high activity of these chloroquine analogues and their mild-to-low toxicity toward host cells make them promising leads for the discovery of new antileishmanial agents.

The mediator role of emotion regulation processes on infertility-related stress.

The objective of this study is to investigate gender differences regarding the mediator role of self-compassion and self-judgment on the effects of external shame, internal shame, dyadic adjustment, on infertility-related stress. One hundred and sixty-two women and 147 men with a primary infertility diagnosis completed the following set of self-report measures: Others as Shamer, Experience of Shame Scale, Dyadic Adjustment Scale, Self-Compassion Scale, and Fertility Problem Inventory. Path analyses results revealed that in women self-compassion fully mediated the effect of internal shame on infertility-related stress and partially mediated the effect of dyadic adjustment on this variable, while external shame had only a direct effect. In men self-judgment fully mediated the effect of external and internal shame on infertility-related stress. Dyadic adjustment had only a direct effect on infertility-related stress. In conclusion, there is a distinct role of self-compassion and self-judgment on the relationship between shame and infertility-related stress in men and women. Such differences should be taken into account in psychological interventions with these patients. Future research is warranted to further support our results.

Fatal case of imported human rabies in Amadora, Portugal, August 2011.

We report on a case of imported human rabies in Portugal, in July 2011 in a woman who presented initially complaining of back pain, without relating exposure to animal bites. She had travelled from Portugal to Bissau, Guinea-Bissau, in April where she had been bitten by a dog on 1 May. She was diagnosed with rabies on 26 July and died two weeks later in spite of being treated following the Milwaukee protocol.

The impact of shame and self-judgment on psychopathology in infertile patients.

BACKGROUND: Little is known about psychological processes of infertile couples pursuing medical treatment in comparison with fertile couples and adoption candidates who also suffer from infertility but are not seeking medical help. This study aims to contribute to a better understanding of these individuals in terms of psychological processes (such as external shame, internal shame and self-judgment) and their association with psychopathology, also attending to gender differences. METHODS: One hundred control couples without known fertility problems [fertile group (FG)], 100 couples with an infertility diagnosis and pursuing medical treatment [infertile group (IG)], and 40 couples with an infertility diagnosis who are applying for adoption [adoption group (AG)] completed the instruments: Beck Depression Inventory, Spielberger State-Trait Anxiety Inventory, Others as Shamer, Experience of Shame Scale and the Self-compassion Scale. One-way analysis of variances were used to compare the three groups demographic and study variables. Pearson correlations and linear multiple regression analysis were performed to investigate the associations between shame, self-judgment, depression and anxiety. To explore gender differences, T-tests were used. RESULTS: The IG group scored higher than FG and AG in measures of depression, anxiety, external shame, internal shame and self-judgment. In infertile couples, self-judgment, external shame and internal shame emerged as significant predictors of depressive symptomatology. Women with an infertility diagnosis tend to present higher levels of depressive and anxiety symptoms in comparison with fertile controls and adoption candidates. CONCLUSIONS: This study highlights the importance of emotional regulation processes such as internal and external shame, and self-judgment, to the understanding of psychopathological symptomatology associated with infertility. Our results suggest that these issues should be addressed in a therapeutic context with these couples. Nevertheless, the heterogeneity of the infertile group, in what concerns different stages of medical diagnosis and treatment, might represent a limitation in the interpretation of our findings.

Candidate gene region for control of rib eye area in Canchim beef cattle.

Investigation of molecular marker effects on production traits is essential to define marker assisted selection strategies in beef cattle. We looked for a possible association of molecular markers and backfat thickness (BFT) and rib eye area (REA) in Canchim (5/8 Charolais + 3/8 Zebu) and MA (offspring of Charolais bulls and 1/2 Canchim + 1/2 Zebu cows) animals raised exclusively on pasture. Traits were measured on 987 individuals from seven herds from two Brazilian States (São Paulo and Goiás), in March and April from 2005 to 2007, when animals were, on average, 19 months of age. Five microsatellite markers lying in QTL regions for BFT and REA (BMS490 and ETH10 on chromosome 5, INRA133 and ILSTS090 on chromosome 6, and BMS2142 on chromosome 19) were genotyped and association analyses were performed under an animal model using the restricted maximum likelihood method. After correction for multiple tests, a significant effect of microsatellite BMS490 on REA was observed, suggesting that at least one QTL affecting carcass traits in this region of the BTA5. No significant effect on BFT was observed for these markers.

Addressing the role of the extrusion pump-bearing pGRT1 plasmid in toluene biodegradation by Pseudomonas putida DOT-T1E under real case scenarios.

The role of both the plasmid pGRT1 and the solvent extrusion pump ttgGHI during toluene biodegradation in Pseudomonas putida DOT-T1E cultures was investigated in a sterile suspended growth bioreactor operated as chemostat and inoculated independently with a wild type strain, a mutant lacking the pGRT1 plasmid (P. putida DOT-TIE-100), and a mutant with a modified pGRT1 plasmid lacking the genes encoding the ttgGHI solvent efflux pump (P. putida DOT-TIE-28). A similar process performance was recorded in all tested strains at 4 g tol m(-3) and dilution rates (D) of 0.1 h(-1). However, operation at 10 g tol m(-3) and D of 0.2 h(-1) revealed a much lower toluene EC (285 g m(-3) h(-1)) in P. putida DOT-T1E-100 cultures when compared to wild type and P. putida DOT-T1E-28 cultures (483-498 g m(-3) h(-1)), which suggests that other mechanisms rather than solvent extrusion by the ttgGHI efflux pump supported this superior process performance. Finally, the plasmid pGRT1 analysed exhibited a remarkable stability towards toluene harmful mediated effects, regardless the strain or toluene loading tested.

Applying modern error theory to the problem of missed injuries in trauma.

BACKGROUND: Modern theory of human error has helped reduce the incidence of adverse events in commercial aviation. It remains unclear whether these lessons are applicable to adverse events in trauma surgery. Missed injuries in a large metropolitan surgical service were prospectively audited and analyzed using a modern error taxonomy to define its applicability to trauma. METHODS: A prospective database of all patients who experienced a missed injury during a 6-month period in a busy surgical service was maintained from July 2006. A missed injury was defined as one that escaped detection from primary assessment to operative exploration. Each missed injury was recorded and categorized. The clinical significance of the error and the level of physician responsible was documented. Errors were divided into planning or execution errors, acts of omission or commission, or violations, slips, and lapses. RESULTS: A total of 1,024 trauma patients were treated by the surgical services over the 6-month period from July to December 2006 in Pietermaritzburg. Thirty-four patients (2.5%) with missed injuries were identified during this period. There were 29 men and 5 women with an average age of 29 years (range: 21-67 years). In 14 patients, errors were related to inadequate clinical assessment. In 11 patients errors involved the misinterpretation of, or failure to respond to radiological imaging. There were 9 cases in which an injury was missed during surgical exploration. Overall mortality was 27% (9 patients). In 5 cases death was directly attributable to the missed injury. The level of the physicians making the error was consultant surgeon (4 cases), resident in training (15 cases), career medical officer (2 cases), referring doctor (6 cases). CONCLUSIONS: Missed injuries are uncommon and are made by all grades of staff. They are associated with increased morbidity and mortality. Understanding the pattern of these errors may help develop error-reduction strategies. Current taxonomies help in understanding the error process, but efforts must be made to develop innovative mechanisms that reduce the potential for error.

Intraflagellar transport complex in Leishmania spp. In silico genome-wide screening and annotation of gene function.

Flagella are constructed and maintained through the highly conserved process of intraflagellar transport (IFT), which is a rapid movement of particles along the axonemal microtubules of cilia/flagella. Particles that are transported by IFT are composed of several protein subunits comprising two complexes (A and B), which are conserved among green algae, nematodes, and vertebrates. To determine whether or not homologues to members of the IFT complex proteins are conserved in Leishmania spp, we scanned genomes, transcriptomes and proteomes of Leishmania species in a search for putative IFT factors, which were then identified in silico, compared, cataloged, and characterized. Since a large proportion of newly identified genes in L. major remain unclassified, with many of these being potentially Leishmania- (or kinetoplastid-) specific, there is a need for detailed analyses of homologs/orthologs that could help us understand the functional assignment of these gene products. We used a combination of integrated bioinformatics tools in a pathogenomics approach to contribute to the annotation of Leishmania genomes, particularly regarding flagellar genes and their roles in pathogenesis. This resulted in the formal in silico identification of eight of these homologs in Leishmania (IFT subunits, 20, 27, 46, 52, 57, 88, 140, and 172), along with others (IFTs 71, 74/72, and 81), as well as sequence comparisons and structural predictions. IFT, an important flagellar pathway in Leishmania, begins to be revealed through screening of trypanosomatid genomes; this information could also be used to better understand fundamental processes in Leishmania, such as motility and pathogenesis.

Apoptosis in alpha interferon (IFN-alpha) intratumoral chemotherapy for cystic craniopharyngiomas.

OBJECTIVES: The aim of this study was to verify whether intracystic injections of alpha-Interferon (IFN-alpha) in cystic craniopharyngiomas were able to reduce the tumor by activating the Fas apoptotic pathway. MATERIALS AND METHODS: Twenty-one patients with cystic craniopharyngiomas treated at the Pediatric Oncology Institute (IOP/GRAACC) of Federal University of São Paulo were submitted to intracystic chemotherapy with IFN-alpha. The tumor sizes of all patients were monitored and the apoptotic factor soluble FasL (sFasL) concentration was determined by ELISA (enzyme-linked immunosorbent assay) in tumor fluid samples from eight patients. RESULTS: There was a complete reduction in 11 patients, a partial response in seven, and a minor response in three patients. The concentration of sFasL was increased in all the eight patients examined concomitantly with the tumor size reduction. CONCLUSIONS: Our data demonstrated that the IFN-alpha was able to induce Fas-mediated apoptosis together with a reduction in the tumor size; such an observation may suggest the importance to investigate still unexplored mechanisms to be exploited in craniopharyngioma therapy.