Postural stability measures in healthy miniature Dachshunds obtained using a pressure mat and a force platform: a validity and reliability study.

BACKGROUND: Miniature Dachshunds have a high prevalence of neurological and musculoskeletal diseases potentially affecting their balance. The postural stability of dogs in quiet standing is an indicator of postural control and can aid in diagnosing and monitoring lameness and other pathologies affecting balance. Measures of centre of pressure (CoP) can be obtained from force and pressure platform systems to evaluate postural stability, however the two systems have not been compared and the latter has not been validated in dogs. The aims of this study were to assess the validity and reliability of using a pressure mat compared to a force platform and report normative values of CoP measures in healthy miniature Dachshunds. Forty two healthy miniature Dachshunds of smooth, long and wire-haired breed types stood still on a pressure mat (Tekscan MatScan®) placed on a force platform and the two systems were synchronised. Maximum anterior-posterior (AP) and medial-lateral (ML) ranges, sway path and 95% area of a best-fit ellipse were computed. Bland-Altman plots and coefficients of correlation assessed validity; intra-class correlation coefficients (ICC) assessed inter-test reliability for both systems. Non-linear regression analyses were used to describe the relationship between CoP and demographic measures. RESULTS: Strong correlations for AP range, ML range and 95% ellipse area and moderate correlation for sway path were found between the two devices. ICC showed good reliability (0.75-0.90) for AP range and moderate (0.5-0.75) for ML range and the 95% ellipse area for both devices. Sway path reliability was excellent (> 0.90) with the force platform but moderate with the pressure mat. Age was positively correlated with balance (inversely correlated with all measures except sway path), while weight explained 94% (force platform) and 27% (pressure mat) of the variance in sway path. CONCLUSIONS: Pressure mats can be used to obtain valid and reliable measures of CoP and replace use of force platforms. Older (non-senior) and heavier (non-obese) dogs show better postural stability. Clinical examinations should include the use of a range of CoP measures when assessing postural balance, while accounting for the effects of age and body weight.

Immunophenotypic changes in juvenile myelomonocytic leukaemia after treatment with hypomethylating agent: Do they help to evaluate dept of response?

5-Azacitidine has been used before stem cell transplantation in juvenile myelomonocytic leukaemia (JMML) patients. Recently, we have described immunophenotypic features in JMML at diagnosis. Here, our aim was to examine the changes in the immunophenotypic features during azacitidine treatment, correlating it with clinical response. Patients treated with 5-azacitidine were evaluated at diagnosis and after three and six cycles of medication. Among 32 patients entering the study, 28 patients were examined after three cycles and 25 patients after six. Patients showed a reduction in CD34/CD117+ cells: median 3.35% at diagnosis, 2.8% after three cycles and 1.63% after six. B-cell progenitors were decreased at diagnosis and decreased after treatment. Monocytes decreased: 11.91% to 6.4% and 4.18% respectively. Complete response was associated with increase in classical monocytes. T lymphocytes, reduced at diagnosis, increased in patients responding to 5-azacitidine. Immunophenotypic aberrancies including expression of CD7 in myeloid progenitors remained after treatment. This feature was associated with a worse response to treatment, as well as presence of NF1. Immunophenotyping was feasible in all patients. Clinical response was associated with a decrease of myeloid progenitors and monocytes and a rise in T lymphocytes although phenotypic aberrancies persisted. The largest effect was observed after three cycles.

Immunophenotypic characteristics of juvenile myelomonocytic leukaemia and their relation with the molecular subgroups of the disease.

The diagnosis of juvenile myelomonocytic leukaemia (JMML) is based on clinical, laboratory and molecular features but immunophenotyping [multiparametric flow cytometry (MFC)] has not been used routinely. In the present study, we describe the flow cytometric features at diagnosis with special attention to the distribution of monocytic subsets and the relation between MFC and molecular subgroups. MFC was performed with an eight-colour platform based on Euroflow. We studied 33 JMML cases. CD34+ /CD117+ /CD13+ cells >2% was found in 25 cases, and 51·5% presented an aberrant expression of CD7. A decrease of CD34+ /CD19+ /CD10+ cells was seen in eight cases and in four they were absent. The granulocytic population had a decreased side scatter in 29 cases. Bone marrow monocytic precursors were increased in 28 patients, with a decrease in classical monocytes (median 80·7%) and increase in CD16+ (intermediate and non-classical). A more pronounced increase in myeloid CD34+ cells was seen in patients with Neurofibromatosis type 1 (NF1) and tyrosine-protein phosphatase non-receptor type 11 (PTPN11), with aberrant CD7 expression in four of six and 10/12 patients respectively. Thus, JMML shows an immunophenotypic profile similar to myelodysplastic syndromes, and a different monocyte subset distribution when compared with chronic MML. MFC proved to be an important diagnostic tool that can help in differential diagnosis with other clonal diseases with monocytosis.

Potent anti-inflammatory activity of pyrenocine A isolated from the marine-derived fungus Penicillium paxilli Ma(G)K.

Very little is known about the immunomodulatory potential of secondary metabolites isolated from marine microorganisms. In the present study, we characterized pyrenocine A, which is produced by the marine-derived fungus Penicillium paxilli Ma(G)K and possesses anti-inflammatory activity. Pyrenocine A was able to suppress, both pretreatment and posttreatment, the LPS-induced activation of macrophages via the inhibition of nitrite production and the synthesis of inflammatory cytokines and PGE2. Pyrenocine A also exhibited anti-inflammatory effects on the expression of receptors directly related to cell migration (Mac-1) as well as costimulatory molecules involved in lymphocyte activation (B7.1). Nitrite production was inhibited by pyrenocine A in macrophages stimulated with CpG but not Poly I:C, suggesting that pyrenocine A acts through the MyD88-dependent intracellular signaling pathway. Moreover, pyrenocine A is also able to inhibit the expression of genes related to NF κ B-mediated signal transduction on macrophages stimulated by LPS. Our results indicate that pyrenocine A has promissory anti-inflammatory properties and additional experiments are necessary to confirm this finding in vivo model.

Impact of HIV-Related Immune Impairment of Yellow Fever Vaccine Immunogenicity in People Living with HIV-ANRS 12403.

The yellow fever (YF) vaccine is one of the safest and most effective vaccines currently available. Still, its administration in people living with HIV (PLWH) is limited due to safety concerns and a lack of consensus regarding decreased immunogenicity and long-lasting protection for this population. The mechanisms associated with impaired YF vaccine immunogenicity in PLWH are not fully understood, but the general immune deregulation during HIV infection may play an important role. To assess if HIV infection impacts YF vaccine immunogenicity and if markers of immune deregulation could predict lower immunogenicity, we evaluated the association of YF neutralization antibody (NAb) titers with the pre-vaccination frequency of activated and exhausted T cells, levels of pro-inflammatory cytokines, and frequency of T cells, B cells, and monocyte subsets in PLWH and HIV-negative controls. We observed impaired YF vaccine immunogenicity in PLWH with lower titers of YF-NAbs 30 days after vaccination, mainly in individuals with CD4 count <350 cells/mm3. At the baseline, those individuals were characterized by having a higher frequency of activated and exhausted T cells and tissue-like memory B cells. Elevated levels of those markers were also observed in individuals with CD4 count between 500 and 350 cells/mm3. We observed a negative correlation between the pre-vaccination level of CD8+ T cell exhaustion and CD4+ T cell activation with YF-NAb titers at D365 and the pre-vaccination level of IP-10 with YF-NAb titers at D30 and D365. Our results emphasize the impact of immune activation, exhaustion, and inflammation in YF vaccine immunogenicity in PLWH.

Effect of Foot Reflexology on Muscle Electrical Activity, Pressure, Plantar Distribution, and Body Sway in Patients with Type 2 Diabetes Mellitus: A Pilot Randomized Controlled Trial.

OBJECTIVE: To verify the effect of foot reflexology on the electrical muscle activity of the lateral and medial gastrocnemius muscle, and to examine the distribution, plantar pressure, and body sway in patients with type 2 diabetes mellitus. METHODS: This pilot randomized controlled trial enrolled 17 volunteers who were clinically diagnosed with diabetes mellitus. The sample was assigned to one of two groups: the control group (CG, n = 7), who received information on foot care and health, and the intervention group (IG, n = 10), who received the application of foot reflexology on specific areas of the feet, for 10 consecutive days. There was blinding of the evaluator and the therapist. Surface electromyography (EMG) was used to assess the electrical activity of the medial and lateral gastrocnemius muscles in maximum voluntary isometric contraction (MVIC) and isotonic contraction (IC); baropodometry and stabilometry were used to analyze unloading, plantar weight distribution, and body sway. RESULTS: There was a statistically significant difference for the variables of maximum peak electrical activity of the left medial gastrocnemius (p = 0.03; effect size = 0.87 and power = 0.81) and left lateral gastrocnemius muscles (p = 0.04, effect size = 0.70 and power = 0.66) respectively, in the intragroup IC, and median frequency of the left medial gastrocnemius muscle in the intragroup MVIC (p = 0.03; effect size = 0.64 and power = 0.59), and in the variables intergroups of the total area on the right side (p = 0.04; effect size = 1.03 and power = 0.50) and forefoot area on the left side (p = 0.02; effect size = 0.51 and power = 0.16). CONCLUSIONS: We conclude that foot reflexology influenced some variables of the intergroup plantar distribution and intragroup EMG in the sample studied. There is a need for a placebo group, a larger sample and a follow-up to strengthen the findings of these experiments.

The activation of D2-like receptors by intranasal dopamine facilitates the extinction of contextual fear and prevents conditioned fear-induced antinociception.

Fear extinction (FExt) is used to treat patients with posttraumatic stress disorder (PTSD). However, fear related to traumatic events can be persistent and return even after successful extinction. The neurochemical control of extinction seems to be performed by several neurotransmitters, including dopamine (DA), through D1 and D2 receptors. Recently, we showed that intranasally applied DA (IN-DA) facilitated the FExt, but the mechanisms by which it promoted this effect are still unknown. This study focused on investigating whether these effects are mediated by the action of DA on D2-like receptors since these receptors seem to be related to neurochemical and molecular changes underlying extinction. Also, we investigated whether IN-DA treatment would affect conditioned fear-induced antinociception (Fear-IA). Rats treated with IN-DA (1 mg/kg) twenty-five minutes after sulpiride (SUL; 40 mg/kg, i.p., D2-antagonist) were subjected to the extinction of contextual fear. IN-DA applied before the extinction session induced the FExt and prevented Fear-IA. These effects were impaired by pre-treatment with SUL, suggesting that the IN-DA effects are mediated by DA on D2-like receptors. SUL per se also facilitated the FExt but did not affect Fear-IA. These data suggest IN-DA as a promising pharmacological tool to supplement the psychotherapy of patients suffering from PTSD.

Pharmacokinetic study of AmB-NP-GR: A new granule form with amphotericin B to treat leishmaniasis and fungal infections.

Amphotericin B (AmB) has been the gold standard to treat systemic fungal infections. The use of AmB is restricted to hospitals because it poses several risks, mainly risks related to its high nephrotoxicity. Given the importance of this drug in medicine, new therapeutics and AmB formulations with nanotechnological improvements are required and could bring many benefits to patients. A new drug formulation with gastro-resistant coated granules has been proposed. The lipid-based system containing AmB was produced and used as raw material in the granulation/coated process. The new developed formulation (AmB-NP-GR) was characterized by optical microscopy, granulometry, and atomic force microscopy (AFM) after disintegration test. AmB-NP-GR showed granular shape, with most granules measured between 250 and 500 µm (37 ± 7% w/w). The AFM images indicated that the granule formulation should disintegrate in the intestine, to release the lipid-based carriers, making them available for absorption and allowing them to reach the blood circulation. The developed formulation was administered to rats in a single dose of 4.0 or 8.0 mg kg-1 and the pharmacokinetics was studied. The samples were analyzed by liquid chromatography coupled to mass spectrometry. Before the pharmacokinetic studies were conducted, the bioanalytical method was validated according to the EMA guideline and all evaluated parameters agreed with this guideline. The pharmacokinetic results showed that Cmax was similar for both doses and that tmax was reached at 4-12 h for dose of 4.0 mg kg-1 and 4 h for dose of 8.0 mg kg-1. The half-life related to the dose of 8.0 mg kg-1 increased significantly compared to the dose of 4.0 mg kg-1 (an increase of more than 3 times). In addition, the mean residence time related to the dose of 8.0 mg kg-1 was 4 times higher than for the lower dose. The clearance value showed to be higher for the lower dose. Together, these results provide important conclusions for experimental design of other in vivo safety and efficacy studies of AmB-NP-GR.

Behavior and illusions: a model to study superstition in a participant replacement experiment.

The notion of superstitious behavior can provide a basic background for understanding such notions as illusions and beliefs. The present study investigated the social mechanism of the transmission of superstitious behavior in an experiment that utilized participant replacement. The sample was composed of a total of 38 participants. Participants performed a task on a computer: they could click a colored rectangle using the mouse. When the rectangle was in a particular color, the participants received points independently of their behavior (variable time schedule). When the color of the rectangle was changed, no points were presented (extinction). Under an Individual Exposure condition, ten participants worked alone on the task. Other participants were exposed to the same experimental task under a Social Exposure condition, in which each participant first learned by observation and then worked on the task in a participant replacement (chain) procedure. The first participant in each chain in the Social Exposure condition was a confederate who worked on the task "superstitiously," clicking the rectangle when points were presented. Superstitious responding was transmitted because of the behavior of the confederate. This also influenced estimates of personal control. These findings suggest that social learning can facilitate the acquisition and maintenance of superstitious behavior and the illusion of control. Our data also suggest that superstitious behavior and the illusion of control may involve similar learning principles.

Establishment of a primary culture of polymorphous low grade adenocarcinoma cells.

OBJECTIVE: The aim of the present study was to establish a primary cell culture derived from polymorphous low grade adenocarcinoma (PLGA). DESIGN: The neoplastic cells were derived from a 57-year-old female patient diagnosed with PLGA. A fragment of the tumor was collected and submitted to enzymatic digestion followed by centrifugation on a Percoll gradient. The cell population was characterized by means of immunofluorescence and detection of PRKD1 gene mutations. RESULTS: Epifluorescence analysis of the primary culture revealed that the malignant epithelial cells were predominantly polygonal in shape and positive for cytokeratin 7, vimentin, and S100. The doubling time of the cell culture was 86.73h. The restriction digestion assay showed that the neoplastic cells possess PRKD1 gene mutations. CONCLUSION: The establishment of primary cell culture derived from PLGA should be considered a useful tool for molecular analysis of this salivary gland tumor.

PrevTev: construção e validação de aplicativo móvel para orientações sobre tromboembolismo venoso / PrevTev: construction and validation of a mobile application for guidelines on venous thromboembolism

Resumo: Introdução: Como o tromboembolismo venoso (TEV) é uma das principais causas de mortes evitáveis em pacientes hospitalizados, é importante a criação de medidas para sua prevenção e seu tratamento. Porém, para a profilaxia de qualidade, a educação e o envolvimento dos pacientes são necessários, e uma das ferramentas para facilitar as atividades educativas em saúde são as tecnologias educacionais. Objetivo: Este estudo teve como objetivos construir e validar um aplicativo móvel sobre orientações para prevenção de TEV em pacientes hospitalizados durante a internação e após a alta hospitalar. Método: Trata-se de um estudo metodológico composto por duas fases (construção e validação), divididas em nove etapas. A fase de validação foi realizada com 11 juízes especialistas e 30 representantes do público-alvo. Utilizaram-se quatro instrumentos de coletas de dados: Suitability Assessment of Materials (SAM), Instrumento de Validação de Conteúdo Educativo em Saúde (IVCES), Instrumento de Avaliação para o Público-alvo e System Usability Scale (SUS), todos interpretados mediante avaliação específica. Resultado: Entre os juízes especialistas, a média da pontuação do SAM foi 89,4%, e a do IVCES, 93,8%. Em relação à opinião do público-alvo, a média de concordância no primeiro instrumento de coleta de dados foi 95%, e a do SUS, 79,5 pontos. Conclusão: O aplicativo móvel foi considerado válido mediante a metodologia aplicada por juízes especialistas e público-alvo.

Abstract: Introduction: Venous thromboembolism (TEV) is one of the main causes of preventable deaths in hospitalized patients. Therefore, it is important to create methods for its prevention and treatment. However, for quality prophylaxis, it is necessary to educate and involve patients, and one of the tools to facilitate educational activities in health is educational technologies. Objective: To build and validate a mobile application on guidelines for preventing TEV in hospitalized patients during hospitalization and after discharge from hospital. Methods: Methodological study consisting of two phases (construction and validation), divided into nine steps. The validation phase was carried out with 11 expert judges and 30 representatives of the target audience. Four data collection instruments were used: Suitability Assessment of Materials (SAM), Health Educational Content Validation Instrument (IVCES), Target Audience Assessment Instrument and System Usability Scale (SUS), all interpreted by specific evaluation. Results: Among the expert judges, the average score for the SAM was 89.4% and for the IVCES, 93.8%. In relation to the opinion of the target audience, the average agreement score in the first data collection instrument was 95% and for the SUS, 79.5 points. Conclusion: The mobile application was considered valid according to the methodology applied by the expert judges and target audience.

Hypusine modification of the ribosome-binding protein eIF5A, a target for new anti-inflammatory drugs: understanding the action of the inhibitor GC7 on a murine macrophage cell line.

Inflammation is part of an important mechanism triggered by the innate immune response that rapidly responds to invading microorganisms and tissue injury. One important elicitor of the inflammatory response is the Gram-negative bacteria component lipopolysaccharide (LPS), which induces the activation of innate immune response cells, the release of proinflammatory cytokines, such as interleukin 1 and tumor necrosis factor α(TNF-α), and the cellular generation of nitric oxide (NO) by the inducible nitric oxide synthase (iNOS). Although essential to the immune response, uncontrolled inflammatory responses can lead to pathological conditions, such as sepsis and rheumatoid arthritis. Therefore, identifying cellular targets for new anti-inflammatory treatments is crucial to improving therapeutic control of inflammation-related diseases. More recently, the translation factor eIF5A has been demonstrated to have a proinflammatory role in the release of cytokines and the production of NO. As eIF5A requires and essential and unique modification of a specific residue of lysine, changing it to hypusine, eIF5A is an interesting cellular target for anti-inflammatory treatment. The present study reviews the literature concerning the anti-inflammatory effects of inhibiting eIF5A function. We also present new data showing that the inhibition of eIF5A function by the small molecule GC7 significantly decreases TNF-α release without affecting TNF-α mRNA levels. We discuss the mechanisms by which eIF5A may interfere with TNF-α mRNA translation by binding to and regulating the function of ribosomes during protein synthesis.

Behavior and illusions: a model to study superstition in a participant replacement experiment

Abstract The notion of superstitious behavior can provide a basic background for understanding such notions as illusions and beliefs. The present study investigated the social mechanism of the transmission of superstitious behavior in an experiment that utilized participant replacement. The sample was composed of a total of 38 participants. Participants performed a task on a computer: they could click a colored rectangle using the mouse. When the rectangle was in a particular color, the participants received points independently of their behavior (variable time schedule). When the color of the rectangle was changed, no points were presented (extinction). Under an Individual Exposure condition, ten participants worked alone on the task. Other participants were exposed to the same experimental task under a Social Exposure condition, in which each participant first learned by observation and then worked on the task in a participant replacement (chain) procedure. The first participant in each chain in the Social Exposure condition was a confederate who worked on the task "superstitiously," clicking the rectangle when points were presented. Superstitious responding was transmitted because of the behavior of the confederate. This also influenced estimates of personal control. These findings suggest that social learning can facilitate the acquisition and maintenance of superstitious behavior and the illusion of control. Our data also suggest that superstitious behavior and the illusion of control may involve similar learning principles.