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Prevention and treatment protocols for taste changes observed during hematopoietic cell transplantation (HCT) are not well-established. The purpose of this study was to assess the efficacy of photobiomodulation (PBM) in relieving taste changes and preventing lingual papillae atrophy. HCT patients received PBM (n = 42) on the tongue dorsum using an InGaAIP laser (660 nm, 100 mW, 1.1 W/cm 2 , 8.8 J/cm 2 ). During the HCT conditioning (T0), severe neutropenia (T1), and after neutrophil engraftment (T2), taste acuity for sweet, bitter, sour, and salty solutions, and clinical appearance of lingual papillae were compared with those of a placebo group (n = 43). PBM significantly reduced hypogeusia, ageusia, and parageusia at T1 and T2, and also successfully prevented papillae atrophy during all the analyzed HCT periods. In conclusion, PBM enhanced taste acuity during HCT. The decrease in papillae atrophy indicated a potential regenerative effect of this therapy on tongue mucosa., (© 2024 Wiley‐VCH GmbH.)

During the state of immune vulnerability in hematopoietic stem cell transplantation (HSCT), the patient has an increased risk of developing a vast number of complications, including severe problems in the oral cavity. These situations require professional oral care to act in the diagnosis and treatment of these conditions, as well as to develop prevention protocols to minimize patient's complications. Oral mucositis, opportunistic infections, bleeding, specific microbiota, taste, and salivary alterations are complications that can occur during HSCT and interfere with various aspects, such as pain control, oral intake, nutrition, bacteremia and sepsis, days of hospitalization and morbidity. Several guidelines have been published to address the role of professional oral care during the HSCT, we describe a consensus regarding these recommendations., Competing Interests: Conflicts of interest None., (Copyright © 2023 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier España, S.L.U. All rights reserved.)

Our objective was to determine the effects of feeding 25-hydroxyvitamin D 3 [25(OH)D 3 ], or vitamin D 3 (cholecalciferol) on plasma, mineral, and metabolite concentrations, mineral balance, mineral excretion, rumination, energy balance, and milk production of dairy cows. We hypothesized that supplementing 3 mg/d of 25(OH)D 3 during the prepartum period would be more effective than supplementing vitamin D 3 at the National Research Council (2001) levels to minimize calcium imbalance during the transition period and improve milk production of dairy cows. Forty multiparous, pregnant nonlactating-Holstein cows were enrolled in this study. Body weight, body condition score, parity, and milk yield in the previous lactation (mean ± standard deviation) were 661 ± 59.2, 3.46 ± 0.35, 1.79 ± 0.87, and 33.2 ± 6.43 kg/d, respectively. Cows were enrolled into the blocks (n = 20 for each treatment) at 30 d of the expected day of calving to receive an acidogenic diet (373 g/kg of neutral detergent fiber and 136 g/kg of crude protein, dry matter basis; -110 mEq/kg) associated with the treatments: (1) control (CTRL), vitamin D 3 at 0.625 mg/d (equivalent to 25,000 IU of vitamin D 3 /d) or (2) 25(OH)D 3 at 3 mg/d (equivalent to 120,000 IU of vitamin D 3 /d). All cows were fed with the base ration for 49 d after calving. Blood samples were taken on d 7, 0, 1, 2, 21, and 42, relative to calving. No effect of treatment was observed for prepartum dry matter intake or body condition score. A trend for increase of ionized Ca was observed for the cows fed 25(OH)D 3 , compared with the CTRL, but no effect of treatment was detected for total Ca or total P. Feeding 25(OH)D 3 increased colostrum yield. The plasmatic concentration of 25-hydroxyvitamin D 3 was increased with 25(OH)D 3 supplementation. 25-Hydroxyvitamin D 3 supplementation increased plasma glucose concentration at parturition. The postpartum dry matter intake was not influenced by treatments. Feeding 25(OH)D 3 increases milk yield, 3.5% fat-corrected milk, and energy-corrected milk and improves milk yield components in early lactation. Overall, these findings suggest that 25(OH)D 3 at 3 mg/d can improve the energy metabolism and lactation performance, compared with the current-feeding practice of supplementing vitamin D 3 at 0.625 mg/d., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Research on olfactory stimulation indicates that it can influence human cognition and behavior, as in the perception of facial expressions. Odors can facilitate or impair the identification of facial expressions, and apparently its hedonic valence plays an important role. However, it was also demonstrated that the presentation of happiness and disgust faces can influence the emotional appraisal of odorants, indicating a bilateral influence in this phenomenon. Hence, it's possible that odor influences on emotional categorization vary depending on the intensity of expressions. To investigate this hypothesis, we performed an emotion recognition task using facial expressions of five emotional categories (happiness, fear, disgust, anger and sadness) with ten different intensities. Thirty-five participants completed four blocks of the task, each with a different olfactory condition, and we found that odorants' effects varied according to the facial expressions intensity. Odorants enhanced the Reaction Time (RT) differences between threshold and high-intensity expressions for disgust and fear faces. Also, analysis of the RT means for high-intensity facial expressions revealed that the well-known advantage in recognition of happiness facial expressions, compared to other emotions, was enhanced in the positive olfactory stimulation and decreased in the negative condition. We conclude that olfactory influences on emotional processing of facial expressions vary along intensities of the latter, and the discrepancies of past research in this field may be a result of a bilateral effect in which the odorants influence the identification of emotional faces just as the facial expressions influence the emotional reaction to the odor., Competing Interests: The authors have declared that no competing interests exist.

Background: In haematopoietic cell transplantation (HCT), oral mucositis and xerostomia are related to conditioning-related oxidative stress. The role of salivary antioxidant enzymes in oral toxicity is poorly described. The aim of this study was to verify the association between salivary antioxidant enzymes and oral mucositis and xerostomia in HCT., Design: Saliva from autologous and allogeneic HCT patients (n = 77) was selected before conditioning (T0), during the neutropenia period (T1) and after marrow engraftment (T2). Salivary flow, total salivary proteins, and superoxide dismutase, catalase and glutathione reductase activities were measured., Results: There were no significant differences in salivary flow, total salivary proteins and catalase at the three HCT time points. Glutathione reductase levels were reduced at T1 compared to T0 (P = .013) and T2 (P = .001). Superoxide dismutase levels were increased from T0 to T2 (P = .013). Neither of these enzymes was associated with oral mucositis. Increased superoxide dismutase levels were associated with xerostomia frequency. Levels of this enzyme also showed significant correlation with days of xerostomia in T2 (ρ = .40, P = .002)., Conclusions: Salivary antioxidant enzymes changed before and during early periods after HCT. The increase in salivary superoxide dismutase suggested partial activation of the salivary antioxidant system and was associated with xerostomia., (© 2020 Stichting European Society for Clinical Investigation Journal Foundation. Published by John Wiley & Sons Ltd.)

Ethical issues that arise during the care of a pregnant woman with cancer are challenging to physicians, policymakers, lawyers, and the bioethics community. The main purpose of this scoping review is to summarize existing literature regarding the bioethical dilemmas when a conflict arises in the maternal-fetus dyad, like the one related to cancer and pregnancy outcomes. Moreover, we illustrate the decision-making process of real-life case reports. Published data were searched through the PubMed and Google Scholar databases, as well as in grey literature, using appropriate controlled keywords in English and Portuguese. After identification, screening, eligibility and data extraction from the articles, a total of 50 was selected. There are several established ethical frameworks for conflict resolution and decision-making. Pragmatic theoretical approaches include case-based analysis, the ethics of care, feminist theory, and traditional ethical principlism that scrutinizes the framework of autonomy, justice, beneficence, and non-maleficence. In addition, society and practitioner values could mediate this complex ethical interplay. The physician must balance autonomy and beneficence-based obligations to the pregnant woman with cancer, along with beneficence-based obligations to the fetus. Ethical challenges have received less attention in the literature, particularly before the third trimester of pregnancy. Best, unbiased and balanced information must be granted both to the patient and to the family, regarding the benefits and harms for the woman herself as well as for the fetal outcome. Based on a previously validated method for analyzing and working up clinical ethical problems, we suggest an adaptation of an algorithm for biomedical decision-making in cancer during pregnancy, including recommendations that can facilitate counseling and help reduce the suffering of the patient and her family., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Alpuim Costa, Nobre, de Almeida, Ferreira, Gonçalves, Braga and Pais.)

Background: To determine whether the busulfan (Bu) present in saliva during hematopoietic cell transplantation (HCT) conditioning correlates with oral mucositis and the changes in salivary antioxidant enzymes., Methods: Bu levels in the plasma and saliva of 19 patients who received HCTs were quantified. Salivary flow and salivary superoxide dismutase and catalase activities were measured during HCT. For the toxicity analysis of salivary Bu, an in vitro assay was conducted by exposing human keratinocytes to artificial saliva containing Bu., Results: Plasma and salivary Bu concentrations were very similar (rho = 0.92, P < 0.001). Salivary Bu concentration correlated with the degree of oral mucositis severity (rho = 0.391, P = 0.029) and was inversely proportional to salivary superoxide dismutase and catalase activities (rho = -0.458, P = 0.036; rho = -0.424, P = 0.043, respectively). Cells exposed to Bu-containing saliva had fewer viable cells (P < 0.01) and more apoptotic cells (P = 0.001) than those exposed to non-Bu-containing saliva., Conclusions: Bu found in saliva during HCT conditioning was correlated with severe oral mucositis and the reduction in salivary antioxidative activity. Furthermore, Bu can be toxic to keratinocytes.

Purpose: The aim of this study was to characterize the taste changes and taste bud atrophy observed in the period of neutropenia of HCT and to determine the influence of transplantation toxicity on these changes., Methods: Autologous and allogeneic HCT patients (n = 51) were selected to perform taste acuity tests prior to conditioning (T0) and during neutropenia (T1). The frequency and time duration of oral mucositis, presence of tongue depapillation, and salivary flow rate were also evaluated. Quality of life was assessed using specific questionnaires., Results: We observed a significant increase in hypogeusia (66.6%, p = 0.001) and dysgeusia (21.4%, p = 0.013) at T1, compared with T0. Bitter taste was the most altered, mainly when the patient underwent conditioning with melphalan (OR = 4.47, p = 0.049). Prolonged oral mucositis (≥ 8 days) (OR = 5.62, p = 0.039) and autologous transplantation (OR = 4.08, p = 0.033) were predictive factors for tongue depapillation. Changes in sour taste (OR = 10.70, p = 0.045) and reduced salivary flow (OR = 21.00, p = 0.013) were associated to body weight loss at T1. Taste changes significantly reduced the quality of life at T1, compared with T0., Conclusions: Frequency of hypogeusia was high in the neutropenia period of the HCT. None of the taste changes was determined by oral mucositis, tongue depapillation, or reduced salivary flow, but melphalan conditioning reduced the bitter taste sensation. Loss of body weight and poor quality of life were associated with taste changes and reduced salivary flow. Further studies are necessary to elucidate this association and the risk factors for taste changes in HCT.

Pembrolizumab is a humanized antibody that targets the programmed death-1 receptor expressed in T cells with high selectivity. This therapeutic is of great importance in cancer immunotherapy yet managing the potential immune-related adverse events remains a concern. Here, we report a rare case of mucous membrane pemphigoid in the oral mucosa, upper respiratory tract, and conjunctiva of a patient with ovarian adenocarcinoma without cutaneous manifestation, which persisted even after pembrolizumab discontinuation. A brief review of pembrolizumab-related bullous pemphigoid cases is presented and possible mechanisms underlying these lesions are discussed.

Self-reported anthropometric data in web-based weight loss interventions may be inaccurate. We studied the agreement between online self-reported and measured weight in the course of the POEmaS randomized controlled trial. Measured weight was not different from reported one (-0.4 kg; 95%CI -0.93 to 0.12). 95.6% of the cases were within the limits of agreement (Bland-Altman method). Self-reported weight collected online was accurate, which suggests that interventions and outcomes assessment can rely on these data.

The brain cholinergic system may undergo structural and functional alterations both in human epilepsy and in respective animal models, but the causal relationships between these alterations and epilepsy remain to be established. In this study, we attempted to examine how the inhibition of epilepsy-related cholinergic plasticity may be reflected in seizure susceptibility and/or in the development of chronic epilepsy and its neurological consequences. For this purpose, adult Wistar rats received intrahippocampal injections of low doses of 192-IgG-saporin (SAP) to produce a moderate, but significant loss of septohippocampal cholinergic cells and to suppress their plasticity. Then, animals were treated with kainic acid to induce status epilepticus, which leads to the development of chronic epilepsy later in life. It was found that SAP-pretreatment was associated with longer latency to the onset of status epilepticus and with reduced mortality rate, suggesting that increased activity of septal cholinergic cells may potentiate seizures. Interestingly, months later, a greater percentage of rats with intact septohippocampal cholinergic connections showed spontaneous seizures, when compared to SAP-pretreated rats. Treatment with kainic acid produced death of 40-50% of hippocampal neurons and this effect was not ameliorated by prior cholinergic depletion. Moreover, the kainate induced cognitive deficits were detected in both SAP-pretreated and sham-pretreated groups. These data suggest that seizure-induced plasticity of cholinergic cells may indeed enhance seizure susceptibility and contribute to epileptogenic processes. They do not support the hypothesis that epilepsy-related hypertrophy of cholinergic neurons may potentiate hippocampal cell loss and respective behavioral impairments., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: This article shows an integrative review on the impact that abnormal color vision may have on the daily routine of individuals., Purpose: We followed the PRISMA guidelines for reviews and carried out researches in four databases (Pubmed, Lilacs, Scopus, and Web of Science) using keywords related to the impact of abnormal color vision., Method: Initially, 805 articles were retrieved and after a first filtering stage, we selected 74 articles for a detailed analysis of the abstracts in which it was found that a total of 20 studies were in fact related to the topic of this review. We then read the selected studies in full and those included in the final selection were analyzed and categorized into specific topic groups of findings. Seven categories were created in total: "impact on daily routine activities", "occupational impact", "impact on product choice motivation", "emotional impact", "impact on school or professional qualification", "impact on self-care and health", and "advantages"., Results: From the definition of these categories we could understand that people with some degree of color vision loss face challenges in different aspects of their daily life, especially in their work activities. Still, the amount of research and hence technical support which could be offered to this population is restricted. Additionally, the scarce availability of publications on the topic and the fact that they include very specific groups of people, such as drivers and medical students, allow us to draw only partial conclusions about the all possible impacts yield by such perceptual difference since they observe the impact of the color-vision deficiency in their daily routine from a specific and precise point of view., Conclusions: A broader view of the impact of this problem on the daily life of its carriers is fundamental for implementing strategies that allow such people to be included in all sorts of activities or for the impact of this sensory change to be decreased or treated in a way that would reduce the detrimental impacts.

Busulfan is a major component of chemotherapy conditioning in hematopoietic cell transplantation (HCT). This alkylating agent is highly toxic at myeloablative doses, exposing HCT patients to risks of mortality. Non-myeloablative (NMA) and reduced-intensity conditioning (RIC) using busulfan have shown impaired toxicity. However, the toxicity of NMA/RIC in the digestive tract is poorly described. This study aimed to characterize the mucositis in the oral cavity (OM), oropharynx/esophagus, and gastrointestinal tract derived from conditionings with myeloablative and non-myeloablative doses of busulfan. We retrospectively retrieved clinical data of HCT patients (n = 100) who underwent myeloablative conditioning (MAC) or NMA/RIC with busulfan. Frequency and time duration of mucositis in the oral cavity and oropharynx/esophagus, diarrhea, and prescription of total parenteral nutrition (TPN) and opioids were also collected. OM severity (p = 0.009) and time duration of mucositis in oropharynx/esophagus (p = 0.022) were frequently higher in MAC than NMA/RIC. A myeloablative dose of busulfan was a risk factor for OM grade ≥ 2 (OR = 4.8, p = 0.002) and for mucositis in oropharynx/esophagus ≥ 5 days (OR = 2.64, p = 0.035). A longer duration of mucositis in the oropharynx/esophagus was also associated with an increase in the prescription of opioids (OR = 7.10, p < 0.001).Overall survival (OS) in MAC was significantly higher than that in NMA/RIC (p = 0.017). No variables related to mucositis interfere significantly in OS. In conclusion, myelosuppression in busulfan-based regimens are predisposed to a high risk for severe OM and to prolonged mucositis in the oropharynx/esophagus.

Psidium guajava L. is a plant widely used for food and in folk medicine all over the world. Studies have shown that guava leaves have antifungal properties. In this study, Flavonoid and Tannic fractions were tested to investigate their chemical composition and antifungal potential in vitro.21 compounds in the two fractions, presenting a higher content of phenolic compounds. The antifungal assays were performed against Candida albicans, Candida tropicalis and Candida krusei by microdilution to determine the IC 50 and the cell viability curve. Minimal Fungicidal Concentration(MFC) and the inhibitory effects of the association of the fractions with Fluconazole, as well as the assays used to verify any morphological changes were performed in microculture chambers based on the concentrations from the microdilution. The IC 50 of the isolated fractions and the fractions associated with each other were calculated, varying from 69.29 to 3444.62 μg/mL and the fractions associated with fluconazole varied from 925.56 to 1.57 μg/mL, it was clear that the association of the natural product with the antifungal presented a synergism. The fractions affected pleomorphism capacity and have a potential antifungal activity as they caused fungal inhibition in isolated use, potentiated the action of Fluconazole, reducing its concentration and impeding morphological transition, one of the virulence factors of the genus., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Introduction: Behaviour change is a key point in weight management. Digital health interventions are attractive tools to deliver behaviour interventions for weight loss, due to the potential to reach a large number of people. We aimed to report how the Behaviour Change Wheel (BCW) was used to develop and implement a web platform to promote weight loss in Brazilian adults with overweight and obesity. Moreover, we aimed to describe the first 12 weeks of usage of the platform in a randomized controlled trial., Methods: The BCW framework was used to define intake of fruit/vegetables, ultra-processed products and sweetened beverages, leisure physical activity and sitting time as target behaviours. The BCW components of behaviour-capability, opportunity and motivation were used to make a behaviour diagnosis of the population and BCW second layer oriented the selection of information, goal setting, self-monitoring, feedback, social support and incentives as behaviour techniques. Using these behaviour techniques, a 24-week behaviour intervention delivered by seven different platform functionalities was developed. The platform was tested in a three-arm parallel (basic platform versus enhanced platform versus minimal intervention control group) randomized controlled trial from September 2017 to April 2018. In the present analysis, we classified the platform functionalities according to the BCW behaviour component (capability, opportunity and behaviour) and used descriptive statistics and Spearman correlations to report functionalities usage according to the BCW behaviour component over the first 12 weeks of the trial. The study was approved by the Ethics Committee of the Federal University of Minas Gerais, Brazil and was registered under NCT 03435445., Results: Over the first 12 weeks of the RCT, the 809 participants (619, 76.5% women; mean age 33.7 years, SD 10.3; mean BMI 29.9 kg/m 2 , SD 4.3) were enrolled for use of the basic and enhance versions of the platform. Capability-driven functionalities were accessed by 455 (56.2%) users with median access of 1 (IQR 1-6) times, whereas opportunity-driven platform functionalities were accessed by 592 participants with 8 (IQR 1-27) median access times and motivation-driven functionalities were accessed by 560 (69.2%) participants with 13 (IQR 1-30) median times of access. Spearman correlations between the use of capability and opportunity functionalities, capability and motivation functionalities and opportunity and motivation functionalities were 0.74 (95% CI 0.70-0.77), 0.74 (95% CI 0.70-0.78), 0.89 (95% CI 0.87-0.91), respectively., Discussion: BCW provided a systematic approach to planning, designing and implementing a complex weight loss intervention based on behaviour change. Moreover, it promoted a clear understanding of the relation between platform functionalities and behaviour determinants. The low use of the capability-driven functionalities might have been related to lack of accuracy in the behaviour diagnosis, as well as to implementation issues. The high correlation between the functionalities use suggests that the BCW approach did not determine the platform usage profile., Conclusion: The BCW provided a framework for an evidence-based intervention on weight loss delivered by a web platform. Using the framework led to a clear understanding of the behaviour determinants and their relation to the platform features.

Leprosy is a chronic dermato-neurological disease caused by infection with Mycobacterium leprae. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes Aedes aegypti and Culex quinquefasciatus and the hemiptera Rhodnius prolixus act as leprosy vectors. By means of real-time PCR quantification of M. leprae 16SrRNA, we found that M. leprae remained viable inside the digestive tract of Rhodnius prolixus for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect M. leprae RNA after a similar period of time. Inside the kissing bug Rhodnius prolixus digestive tract, M. leprae was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of Trypanosoma cruzi, a well-known pathogen transmitted by this insect. The maintenance of M. leprae infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that Rhodnius prolixus defecate infective M. leprae, justifying the evaluation of the presence of M. leprae among sylvatic and domestic kissing bugs in countries endemic for leprosy.

Objective: To compare the transtibial and two-incision techniques for anterior cruciate ligament (ACL) reconstruction using a single band., Methods: A prospective and randomized study was conducted in blocks. Patients underwent ACL reconstruction by means of two techniques: transtibial (group 1: 20 patients) or two incisions (group 2: 20 patients). The radiographic positioning of the tunnel, inclination of the graft, graft isometricity and functional results (IKDC and Lysholm) were evaluated., Results: The positioning of the femoral tunnel on the anteroposterior radiograph, expressed as a mean percentage relative to the medial border of the tibial plateau, was 54.6% in group 1 and 60.8% in group 2 (p < 0.05). The positioning of the femoral tunnel on the lateral radiograph, expressed as a mean percentage relative to the anterior border of Blumensaat's line, was 68.4% in group 1 and 58% in group 2 (p < 0.05). The mean inclination of the graft was 19° in group 1 and 27.2° in group 2 (p < 0.05). The mean graft isometricity was 0.96 mm in group 1 and 1.33 mm in group 2 (p > 0.05). Group 2 had better results from the pivot-shift maneuver (p < 0.05)., Conclusion: The technique of two incisions allowed positioning of the femoral tunnel that was more lateralized and anteriorized, such that the graft was more inclined and there was a clinically better result from the pivot-shift maneuver. There was no difference in isometricity and no final functional result over the short follow-up time evaluated.

In mammals, lipid droplets (LDs) are ubiquitous organelles that modulate immune and inflammatory responses through the production of lipid mediators. In insects, it is unknown whether LDs play any role during the development of immune responses. We show that Aedes aegypti Aag2 cells - an immune responsive cell lineage - accumulates LDs when challenged with Enterobacter cloacae, Sindbis, and Dengue viruses. Microarray analysis of Aag2 challenged with E.cloacae or infected with Dengue virus revealed high transcripts levels of genes associated with lipid storage and LDs biogenesis, correlating with the increased LDs numbers in those conditions. Similarly, in mosquitoes, LDs accumulate in midgut cells in response to Serratia marcescens and Sindbis virus or when the native microbiota proliferates, following a blood meal. Also, constitutive activation of Toll and IMD pathways by knocking-down their respective negative modulators (Cactus and Caspar) increases LDs numbers in the midgut. Our results show for the first time an infection-induced LDs accumulation in response to both bacterial and viral infections in Ae. Aegypti, and we propose a role for LDs in mosquito immunity. These findings open new venues for further studies in insect immune responses associated with lipid metabolism.

Background: Here we report the monitoring of the digestive tract colonization of Rhodnius prolixus by Trypanosoma cruzi using an accurate determination of the parasite load by qPCR coupled with fluorescence and bioluminescence imaging (BLI). These complementary methods revealed critical steps necessary for the parasite population to colonize the insect gut and establish vector infection., Methodology/principal Findings: qPCR analysis of the parasite load in the insect gut showed several limitations due mainly to the presence of digestive-derived products that are thought to degrade DNA and inhibit further the PCR reaction. We developed a real-time PCR strategy targeting the T. cruzi repetitive satellite DNA sequence using as internal standard for normalization, an exogenous heterologous DNA spiked into insect samples extract, to precisely quantify the parasite load in each segment of the insect gut (anterior midgut, AM, posterior midgut, PM, and hindgut, H). Using combined fluorescence microscopy and BLI imaging as well as qPCR analysis, we showed that during their journey through the insect digestive tract, most of the parasites are lysed in the AM during the first 24 hours independently of the gut microbiota. During this short period, live parasites move through the PM to establish the onset of infection. At days 3-4 post-infection (p.i.), the parasite population begins to colonize the H to reach a climax at day 7 p.i., which is maintained during the next two weeks. Remarkably, the fluctuation of the parasite number in H remains relatively stable over the two weeks after refeeding, while the populations residing in the AM and PM increases slightly and probably constitutes the reservoirs of dividing epimastigotes., Conclusions/significance: These data show that a tuned dynamic control of the population operates in the insect gut to maintain an equilibrium between non-dividing infective trypomastigote forms and dividing epimastigote forms of the parasite, which is crucial for vector competence.

Background: Protein Tyrosine Phosphatases (PTPs) are enzymes that catalyze phosphotyrosine dephosphorylation and modulate cell differentiation, growth and metabolism. In mammals, PTPs play a key role in the modulation of canonical pathways involved in metabolism and immunity. PTP1B is the prototype member of classical PTPs and a major target for treating human diseases, such as cancer, obesity and diabetes. These signaling enzymes are, hence, targets of a wide array of inhibitors. Anautogenous mosquitoes rely on blood meals to lay eggs and are vectors of the most prevalent human diseases. Identifying the mosquito ortholog of PTP1B and determining its involvement in egg production is, therefore, important in the search for a novel and crucial target for vector control., Methodology/principal Findings: We conducted an analysis to identify the ortholog of mammalian PTP1B in the Aedes aegypti genome. We identified eight genes coding for classical PTPs. In silico structural and functional analyses of proteins coded by such genes revealed that four of these code for catalytically active enzymes. Among the four genes coding for active PTPs, AAEL001919 exhibits the greatest degree of homology with the mammalian PTP1B. Next, we evaluated the role of this enzyme in egg formation. Blood feeding largely affects AAEL001919 expression, especially in the fat body and ovaries. These tissues are critically involved in the synthesis and storage of vitellogenin, the major yolk protein. Including the classical PTP inhibitor sodium orthovanadate or the PTP substrate DiFMUP in the blood meal decreased vitellogenin synthesis and egg production. Similarly, silencing AAEL001919 using RNA interference (RNAi) assays resulted in 30% suppression of egg production., Conclusions/significance: The data reported herein implicate, for the first time, a gene that codes for a classical PTP in mosquito egg formation. These findings raise the possibility that this class of enzymes may be used as novel targets to block egg formation in mosquitoes.

Objective: This study assessed the anterior-posterior positioning of the upper and lower first molars, and the degree of rotation of the upper first molars in individuals with Class II, division 1, malocclusion., Methods: Asymmetry I, an accurate device, was used to assess sixty sets of dental casts from 27 females and 33 males, aged between 12 and 21 years old, with bilateral Class II, division 1. The sagittal position of the molars was determined by positioning the casts onto the device, considering the midpalatal suture as a symmetry reference, and then measuring the distance between the mesial marginal ridge of the most distal molar and the mesial marginal ridge of its counterpart. With regard to the degree of rotation of the upper molar, the distance between landmarks on the mesial marginal ridge was measured. Chi-square test with a 5% significance level was used to verify the variation in molars position. Student's t test at 5% significance was used for statistical analysis., Results: A great number of lower molars mesially positioned was registered, and the comparison between the right and left sides also demonstrated a higher number of mesially positioned molars on the right side of both arches. The average rotation of the molars was found to be 0.76 mm and 0.93 mm for the right and left sides, respectively., Conclusions: No statistically significant difference was detected between the mean values of molars mesialization regardless of the side and arch. Molars rotation, measured in millimeters, represented ¼ of Class II.

Background: The understanding of mosquito immune responses can provide valuable tools for development of novel mosquito control strategies. Aiming the study at insect innate immunity, continuous insect cell lines have been established and used as research tools due to the fact that they constitute more homogeneous, sensitive, and reproducible systems than the insects from which they originated. More recently, Aag-2, an Aedes aegypti cell lineage, began to be frequently used as a model for studies of mosquito immunity. Nevertheless, to our knowledge, no study has systematically characterized the responses of Aag-2 cell line against different kinds of pathogens and compared its response to those exhibited by whole mosquitoes. For this reason, in this study we characterized gene expression profiles of the Aag-2 cell line in response to different kinds of immune challenges, such as Gram negative and positive bacteria, fungi and viruses, comparing the obtained results with the ones already described in the literature for whole mosquitoes., Methods: Aedes aegypti Aag-2 cells were exposed to different immune stimuli (gram-positive and gram negative heat inactivated bacteria, zymosan or Sindbis virus) for 24 hours and the expression of selected marker genes from toll, IMD and Jak/STAT pathways was analyzed by qPCR. Also, cells were incubated with fluorescent latex beads for evaluation of its phagocytosis capacity., Results: Aag-2 cells were stimulated with two concentrations of heat-killed Gram negative (Enterobacter cloacae) or Gram positive (Micrococcus luteus) bacteria, Zymosan or infected with Sindbis virus and the expression of key genes from the main immune related pathways, Toll, IMD and Jak/STAT, were investigated. Our results suggest that Toll and IMD pathways are activated in response to both Gram positive and negative bacteria and Zymosan in Aag-2 cells, displaying an immune profile similar to those described in the literature for whole mosquitoes. The same stimuli were also capable of activating Jak/STAT pathway in Aag-2 cells. Infection with Sindbis virus led to an up-regulation of the transcription factor STAT but was not able to induce the expression of any other gene from any of the pathways assayed. We also showed that this cell line is able to phagocytose latex beads in culture., Conclusions: Our results characterize the expression profile of Aag-2 cells in response to different immune stimuli and demonstrate that this cell lineage is immune-competent and closely resembles the response described for whole Ae. aegypti mosquitoes. Hence, our findings support the use of Aag-2 as a tool to comprehend Ae. aegypti immune response both at cellular and humoral levels.

This paper proposes a new method for determination of amoxicillin in pharmaceutical suspension formulations, based on transflectance near infrared (NIR) measurements and partial least squares (PLS) multivariate calibration. A complete methodology was implemented for developing the proposed method, including an experimental design, data preprocessing by using multiple scatter correction (MSC) and outlier detection based on high values of leverage, and X and Y residuals. The best PLS model was obtained with seven latent variables in the range from 40.0 to 65.0 mg mL(-1) of amoxicillin, providing a root mean square error of prediction (RMSEP) of 1.6 mg mL(-1). The method was validated in accordance with Brazilian and international guidelines, through the estimate of figures of merit, such as linearity, precision, accuracy, robustness, selectivity, analytical sensitivity, limits of detection and quantitation, and bias. The results for determinations in four commercial pharmaceutical formulations were in agreement with the official high performance liquid chromatographic (HPLC) method at the 99% confidence level. A pseudo-univariate calibration curve was also obtained based on the net analyte signal (NAS). The proposed chemometric method presented the advantages of rapidity, simplicity, low cost, and no use of solvents, compared to the principal alternative methods based on HPLC., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The mosquito Aedes aegypti can spread the dengue, chikungunya and yellow fever viruses. Thus, the search for key molecules involved in the mosquito survival represents today a promising vector control strategy. High Mobility Group Box (HMGB) proteins are essential nuclear factors that maintain the high-order structure of chromatin, keeping eukaryotic cells viable. Outside the nucleus, secreted HMGB proteins could alert the innate immune system to foreign antigens and trigger the initiation of host defenses. In this work, we cloned and functionally characterized the HMGB1 protein from Aedes aegypti (AaHMGB1). The AaHMGB1 protein typically consists of two HMG-box DNA binding domains and an acidic C-terminus. Interestingly, AaHMGB1 contains a unique alanine/glutamine-rich (AQ-rich) C-terminal region that seems to be exclusive of dipteran HMGB proteins. AaHMGB1 is localized to the cell nucleus, mainly associated with heterochromatin. Circular dichroism analyses of AaHMGB1 or the C-terminal truncated proteins revealed α-helical structures. We showed that AaHMGB1 can effectively bind and change the topology of DNA, and that the AQ-rich and the C-terminal acidic regions can modulate its ability to promote DNA supercoiling, as well as its preference to bind supercoiled DNA. AaHMGB1 is phosphorylated by PKA and PKC, but not by CK2. Importantly, phosphorylation of AaHMGB1 by PKA or PKC completely abolishes its DNA bending activity. Thus, our study shows that a functional HMGB1 protein occurs in Aedes aegypt and we provide the first description of a HMGB1 protein containing an AQ-rich regulatory C-terminus.

Recently, we showed that infection with dengue virus increases the locomotor activity of Aedes aegypti females. We speculate that the observed increased locomotor activity could potentially increase the chances of finding a suitable host and, as a consequence, the relative biting rate of infected mosquitoes. We used a mathematical model to investigate the impact of the increased locomotor activity by assuming that this activity translated into an increased biting rate for infected mosquitoes. The results show that the increased biting rate resulted in dengue outbreaks with greater numbers of primary and secondary infections and with more severe biennial epidemics.

The intrinsic yield of spermatogenesis and supporting capacity of Sertoli cells are the desirable indicators of sperm production in a species. The objective of the present study was to quantify intrinsic yield and the Sertoli cell index in the spermatogenic process and estimate testicular sperm reserves by histological assessment of fragments obtained by testicular biopsy of five adult jaguars in captivity. The testicular fragments were fixed in 4% glutaric aldehyde, dehydrated at increasing alcohol concentrations, included into hydroxyethyl methacrylate, and were cut into 4 microm thickness. In the seminiferous epithelium of the jaguar, 9.2 primary spermatocytes in pre-leptotene were produced by "A" spermatogonia. During the meiotic divisions only 3.2 spermatids were produced by a primary spermatocyte. The general spermatogenic yield of the jaguar was about 23.4 cells and each Sertoli cell was able to maintain about 19.2 germ cells, 11 of them were round spermatids. In each seminiferous epithelium cycle about 166 million spermatozoa were produced by each gram of testicular tissue. In adult jaguars, the general spermatogenic yield was similar to the yield observed in pumas, greater than that observed for the domestic cat, but less compared to most domestic animals.

The endocrine portion of mammal testicle is represented by Leydig cells which, together with connective cells, leukocytes, blood and lymphatic vessels, form the intertubular space. The arrangement and proportion of these components vary in the different species of mammals and form mechanisms that keep the testosterone level--the main product of the Leydig cell--two to three times higher in the interstitial fluid than in the testicular blood vessels and 40-250 times higher in these than in the peripheral blood. Marked differences are observed among animal species regarding the abundance of Leydig cells, loose connective tissue, development degree and location of the lymphatic vessels and their topographical relationship with seminiferous tubules. In the jaguar about 13% of the testicular parenchyma is occupied by Leydig cells, 8.3% by connective tissue and 0.3% by lymphatic vessels. Although included in standard II, as described in the literature, concerning the arrangement of the intertubular space, the jaguar has grouped lymphatic vessels in the intertubular space instead of isolated ones. In the jaguar the average volume of the Leydig cell was 2386 microm3 and its average nuclear diameter was 7.7 microm. A great quantity of 2.3 microm diameter lipidic drops was observed in the Leydig cell cytoplasm of the jaguar. The Leydig cells in the jaguar occupy an average 0.0036% of the body weight and the average number per gram of testicle was within the range for most mammals: between 20 and 40 million.

In order to evaluate the prevalence of psychiatric disorders in a Brazilian general hospital and their association with religious denomination and religiosity, 253 inpatients were interviewed. A socio-demographic questionnaire and an instrument for diagnosis of mental disorders (MINI-Plus) were applied. Distribution of religious denominations was: Catholic 63.2% (n=177), Evangelical Protestant 20.4% (n=57), Spiritist 4.3% (n=12), traditional Protestant 2.3% (n=8), and "no religion" 7.5% (n=21). Degree of religiosity was: very religious 43.2% (n=116), religious 46.9% (n=129), hardly religious 9.8% (n=27), and not at all religious 1.1% (n=3). Evangelical (Pentecostal) religious affiliation and frequent attendance at worship services were associated with fewer alcohol problems. Membership in an Evangelical (Pentecostal) church may thus have an inhibitory effect on alcohol dependence or abuse. Intensity of religiosity was moderately associated with overall prevalence of disorders, especially bipolar disorder. It is reasonable to conclude that extreme situations (very intense versus very limited religious participation) are related to this finding, associating both an exacerbated pursuit of religion and alienation from it with altered mental states.

Two proteins from the eggshell of Rhodnius prolixus were isolated, characterized and named Rp30 and Rp45 according to their molecular masses. Purified proteins were used to obtain specific antiserum which was later used for immunolocalization. The antiserum against Rp30 and Rp45 detected their presence inside the follicle cells, their secretion and their association with oocyte microvilli. Both proteins are expressed during the final stage of vitellogenesis, preserved during embryogenesis and discarded together with the eggshell. The amino terminals were sequenced and both proteins were further cloned using degenerated primers. The amino acid sequences appear to have a tripartite arrangement with a highly conserved central domain which presents a repetitive motif of valine-proline-valine (VPV) at intervals of 15 amino acid residues. Their amino acid sequence showed no similarity to any known eggshell protein. The expression of these proteins was also investigated; the results demonstrated that this occurred strictly in choriogenic follicles. Antifungal activity against Aspergillus niger was found to be associated with Rp45 but not with Rp30. A. niger exposed to Rp45 protein induced growth inhibition and several morphological changes such as large vacuoles, swollen mitochondria, multi-lamellar structures and a disorganized cell wall as demonstrated by electron microscopy analysis.

Objective: To estimate the proportion of inpatients at a university general hospital who are at risk of committing suicide., Method: A random sample of 253 patients (57% males) aged 18 years old or older, admitted to surgical and clinical wards, was assessed using the the Mini International Neuropsychiatric Interview, which has a section that evaluates the risk for suicide. Uni- and multivariate analyses were performed., Results: There were 58 (23%) patients with a risk for suicide, 13 (5% of total) of whom presented a high risk. The prevalence of suicide risk was greater in young adult patients, those with no matrimonial relationship and those diagnosed with major depression (univariate analysis, Chi-squared test; p = 0.01, 0.03 and 0.0001, respectively). The multivariate analysis revealed that the risk for suicide in individuals younger than 30 years old was two fold higher than in those individuals between the ages of 30 and 59 years (OR = 0.45, 95% CI = 0.22-0.93; p = 0.03) and four fold greater than in those who were 60 years old or older (OR = 0.25, 95% CI = 0.1-0.64; p = 0.004)., Conclusion: When young adults are admitted to general hospitals they should receive special attention due to their suicidal potential.

Malnutrition is the fourth commonest reason for hospital admission to the paediatric department of the Central Hospital, Maputo and has the second highest death rate (20 per cent). A study from 1995 into mortality at this paediatric department, suggested an increase in severe malnutrition. Recent studies have shown that the global burden of undernutrition in the world is declining; however, data for Eastern Africa shows a deterioration. The current study was aimed at describing and comparing the patients on the malnutrition ward, in 2001 and 1983. The study gathered indices of nutritional status and secondary diagnoses from the notes of all children (aged between 6 months and 5 years) discharged from the malnutrition ward for a period of l year (January-December 2001), and from data (collected in January-December 1983) for the malnutrition ward. Data was entered and analysed using Epi-Info 6 and SPSS statistics package. The ethics committee of the hospital approved the study. Data was collected for 558 children in 2001 and 833 in 1983. There was no gender difference, average age was 21.7 months in 2001 and 23.8 months in 1983 and the average hospital stay was 13.1 and 14.3 days, respectively. In 2001, 33 per cent had kwashiorkor, 26 per cent marasmus, and 28 per cent marasmic kwashiorkor. Three hundred and twenty children (82 per cent) were <2 Z-scores below the median weight-for-age and 252 children (65 per cent) were <3 Z-scores. Forty per cent had malaria, 65 per cent anaemia, 53 per cent bronchopneumonia, 14 per cent TB, 36 per cent diarrhoea, and 12 per cent HIV/AIDS. In 1983, 49 per cent had kwashiorkor, 17 per cent marasmus, and 11 per cent had marasmic kwashiorkor. A total of 494 children (81 per cent) were <2 Z-scores below the median weight-for-age and 335 children (55 per cent) were <3 Z-scores. Eighteen per cent had malaria, 37 per cent anaemia, 28 per cent bronchopneumonia, 6 per cent TB, 8 per cent diarrhoea, and 4.4 per cent measles/post-measles. A comparison between the clinical status of 1983 with that of 2001 shows little difference in age, gender or length of stay. There were fewer admissions in 2001, although a higher percentage of severely underweight children and the 2001 group had more secondary infections, especially malaria, bronchopneumonia and anaemia. Clinical malnutrition at a referral hospital level, in spite of the remarkable Mozambican economic growth, shows signs of following the depressing pattern for much of Eastern Africa. A prospective study including HIV tests and anthropometric data for this and the city's other hospitals is warranted. Discussion needs to be prompted on a local level about malnutrition and the use of guidelines.

Cystic fibrosis is a genetic disease usually diagnosed by abnormal sweat testing. We report a case of an 18-year-old female with bronchiectasis, chronic P. aeruginosa infection, and normal sweat chloride concentrations who experienced rapid decrease of lung function and clinical deterioration despite treatment. Given the high suspicion of cystic fibrosis, broad genotyping testing was performed, showing a compound heterozygous with deltaF508 and 3849+10 kb C-->T mutations, therefore confirming cystic fibrosis diagnosis. Although the sweat chloride test remains the gold standard for the diagnosis of cystic fibrosis, alternative diagnostic tests such as genotyping and electrophysiologic measurements must be performed if there is suspicion of cystic fibrosis, despite normal or borderline sweat chloride levels.

Three cases of patients with transient epileptic amnesia (also known as pure amnestic seizure) are described. In two patients it was the unique seizure type and represented de novo epilepsy occurring in the elderly. In the third patient it coexisted with long standing complex partial seizure of mesial temporal lobe origin. The problems associated with the diagnosis and the main clinical features are discussed. In addition the underlining pathophysiological mechanisms are considered. It is argued that this seizure type is likely to be under-diagnosed and that further research is needed as the presence of TEA has significant implications for clinical and surgical management., (Copyright 2002 Published by Elsevier Science Ltd on behalf of BEA Trading Ltd.)

Objective: This is a study to evaluate friendships in latency street boys of Porto Alegre, RGS, Brazil., Methods: A sample of 30 latency street boys was compared with a sample of 51 latency boys living with their low income families, using the Cornell Interview of Peers and Friends (CIPF)., Results: The two groups had a significantly different CIPF global scores, and the boys of the street group had the highest mean score. Also, boys of the street had significantly lower developmental appropriateness, self-esteem and social skills scores than boys living with a family., Conclusions: The urgent need for intervention street children, especially on boys of the street, is emphasized.