Risk of Cognitive Impairment in Patients With Parkinson’s Disease With Visual Hallucinations and Subjective Cognitive Complaints

Background@#and Purpose Visual hallucinations (VH) and subjective cognitive complaints (SCC) are associated with cognitive impairment (CI) in Parkinson’s disease. Our aims were to determine the association between VH and SCC and the risk of CI development in a cohort of patients with Parkinson’s disease and normal cognition (PD-NC). @*Methods@#Patients with PD-NC (total score of >80 on the Parkinson’s Disease Cognitive Rating Scale [PD-CRS]) recruited from the Spanish COPPADIS cohort from January 2016 to November 2017 were followed up after 2 years. Subjects with a score of ≥1 on domain 5 and item 13 of the Non-Motor Symptoms Scale at baseline (V0) were considered as “with SCC” and “with VH,” respectively. CI at the 2-year follow-up (plus or minus 1 month) (V2) was defined as a PD-CRS total score of <81. @*Results@#At V0 (n=376, 58.2% males, age 61.14±8.73 years [mean±SD]), the frequencies of VH and SCC were 13.6% and 62.2%, respectively. VH were more frequent in patients with SCC than in those without: 18.8% (44/234) vs 4.9% (7/142), p<0.0001. At V2, 15.2% (57/376) of the patients had developed CI. VH presenting at V0 was associated with a higher risk of CI at V2 (odds ratio [OR]=2.68, 95% confidence interval=1.05–6.83, p=0.039) after controlling for the effects of age, disease duration, education, medication, motor and nonmotor status, mood, and PD-CRS total score at V0. Although SCC were not associated with CI at V2, presenting both VH and SCC at V0 increased the probability of having CI at V2 (OR=3.71, 95% confidence interval=1.36–10.17, p=0.011). @*Conclusions@#VH were associated with the development of SCC and CI at the 2-year follow-up in patients with PD-NC.

Motor Fluctuations Development Is Associated with Non-Motor Symptoms Burden Progression in Parkinson's Disease Patients: A 2-Year Follow-Up Study.

Objective: The aim of the present study was to analyze the progression of non-motor symptoms (NMS) burden in Parkinson's disease (PD) patients regarding the development of motor fluctuations (MF). Methods: PD patients without MF at baseline, who were recruited from January 2016 to November 2017 (V0) and evaluated again at a 2-year follow-up (V2) from 35 centers of Spain from the COPPADIS cohort, were included in this analysis. MF development at V2 was defined as a score ≥ 1 in the item-39 of the UPDRS-Part IV, whereas NMS burden was defined according to the Non-motor Symptoms Scale (NMSS) total score. Results: Three hundred and thirty PD patients (62.67 ± 8.7 years old; 58.8% males) were included. From V0 to V2, 27.6% of the patients developed MF. The mean NMSS total score at baseline was higher in those patients who developed MF after the 2-year follow-up (46.34 ± 36.48 vs. 34.3 ± 29.07; p = 0.001). A greater increase in the NMSS total score from V0 to V2 was observed in patients who developed MF (+16.07 ± 37.37) compared to those who did not develop MF (+6.2 ± 25.8) (p = 0.021). Development of MF after a 2-year follow-up was associated with an increase in the NMSS total score (ß = 0.128; p = 0.046) after adjustment to age, gender, years from symptoms onset, levodopa equivalent daily dose (LEDD) and the NMSS total score at baseline, and the change in LEDD from V0 to V2. Conclusions: In PD patients, the development of MF is associated with a greater increase in the NMS burden after a 2-year follow-up.

Prevalence of psychological symptoms and associated risk factors in a Spanish sample of HIV-positive youth compared to uninfected peers.

INTRODUCTION: The aim of the study was twofold: a) to determine the prevalence of symptoms of depression and anxiety and sleep disturbances in young patients with vertically-transmitted HIV infection compared to uninfected peers, and b) to identify sociodemographic, psychosocial and medication-related variables and other clinical risk and protective factors related to psychological symptoms. METHODS: We conducted a cross-sectional study in two groups with independent measures (36 youth with vertically transmitted HIV infection and 39 HIV-negative peers). We used 3 standardised assessment tools and a sociodemographic/psychosocial questionnaire (STAI, BDI, PSQI and adapted sociodemographic test). We performed univariate and multivariable analyses. RESULTS: The univariate analysis did not find significant differences between groups either in psychosocial factors or in the clinical scores. The multivariable analysis found that the presence of psychological symptoms was strongly associated with sociodemographic factors and past events. CONCLUSIONS: Psychosocial factors and the social environment seemed to correlate more strongly to psychological symptoms than HIV status and to explain better the current psychological state of individuals.

Increased homocysteine levels correlate with cortical structural damage in Parkinson's disease.

BACKGROUND: Blood homocysteine appears to be increased in Parkinson's disease (PD) and may play a role in the development and progression of this disorder. However, the specific contribution of abnormal homocysteine levels to cortical degeneration in PD remains elusive. OBJECTIVE: To characterize the cortical structural correlates of homocysteine levels in PD. METHODS: From the COPPADIS cohort, we identified a subset of PD patients and healthy controls (HC) with available homocysteine and imaging data. Surface-based vertex-wise multiple regression analyses were performed to investigate the cortical macrostructural (cortical thinning) and microstructural (increased intracortical diffusivity) correlates of homocysteine levels in this sample. RESULTS: A total of 137 PD patients and 43 HC were included. Homocysteine levels were increased in the PD group (t = -2.2, p = 0.03), correlating in turn with cognitive performance (r = -0.2, p = 0.03). Homocysteine in PD was also associated with frontal cortical thinning and, in a subset of patients with available DTI data, with microstructural damage in frontal and posterior-cortical regions (p < 0.05 Monte-Carlo corrected). CONCLUSIONS: Homocysteine in PD appears to be associated with cognitive performance and structural damage in the cerebral cortex. These findings not only reinforce the presence and importance of cortical degeneration in PD, but also suggest that homocysteine plays a role among the multiple pathological processes thought to be involved in its development.

A real-life overview of a hematopoietic cell transplant program throughout a four-year period, including prospective registry, exclusion causes and final donor selection.

Traceability of patients who are candidates for Hematopoietic cell transplant (HCT) is crucial to ensure HCT program quality. Continuous knowledge of both a detailed registry from a HCT program and final exclusion causes can contribute to promoting a real-life vision and optimizing patient and donor selection. We analyzed epidemiological data reported in a 4 year-monocentric prospective registry, which included all patients presented as candidates for autologous (Auto) and/or allogeneic (Allo) HCT. A total of 543 patients were considered for HCT: 252 (42.4%) for Allo and 291 (57.6%) for Auto. A total of 98 (38.9%) patients were excluded from AlloHCT due to basal disease progression more commonly (18.2%). Seventy-six (30.2%) patients had an HLA identical sibling, whereas 147 (58.3%) patients had only Haplo. UD research was performed in 106 (42%) cases, significantly more often in myeloid than lymphoid malignancies (57% vs 28.7%, p < 0.001) but 61.3% were finally canceled, due to donor or disease causes in 72.4%. With respect to Auto candidates, a total of 60 (20.6%) patients were finally excluded; progression was the most common cause (12%). Currently, Haplo is the most frequent donor type. The high cancellation rate of UD research should be revised to optimize further donor algorithms.

Dietary protein sources and their effects on faecal odour and the composition of volatile organic compounds in faeces of French Bulldogs.

The strong odour of faeces and excessive production of gases in some dog breeds have long been a concern of owners. The pet food industry uses nutritional alternatives, such as high-quality ingredients and additives, to improve the odour of faeces. However, there are still some dog breeds, such as the French Bulldog, that present this problem due to the presence into the large intestine of indigested protein. Therefore, a deeper understanding of the volatile compounds that influence the odour of dog faeces is important. This study aimed to identify changes of faecal odour compounds that are most prevalent in French Bulldogs based on food containing different high-quality protein sources and their effect in sensory analysis. Four maintenance foods with different protein sources were formulated: P, poultry meal food; W, wheat gluten food; PW, poultry meal and wheat gluten food; and PWH, poultry meal, wheat gluten, and hydrolysed protein food. Eight adult French Bulldogs were arranged in a 4x4 Latin square design and adapted to foods for 28 days. Fresh faeces were collected for analysis of volatile organic compounds (VOCs) and sensory analysis. The means were compared by SAS, and statistical significance was indicated by p ≤ 0.05. No adverse effects were observed in the animals regarding VOCs, and a significant difference was observed in two of the 68 compounds identified. The animals fed a P food had higher concentrations of phenol in the faeces, whereas the indole compound was present at higher concentrations in animals fed the W food. P food was associated with higher odour perception during sensory evaluation. In summary, the source of protein in the foods had little impact on the composition of VOCs, and a greater perception of the odour was determined by sensory analysis when foods containing animal protein were administered.

The SARS-CoV-2 receptor-binding domain expressed in Pichia pastoris as a candidate vaccine antigen

1.The effort to develop vaccines based on economically accessible technological platforms available by developing countries vaccine manufacturers is essential to extend the immunization to the whole world population and to achieve the desired herd immunity, necessary to end the COVID-19 pandemic. Here we report on the development of a SARS-CoV-2 receptor-binding domain (RBD) protein, expressed in yeast Pichia pastoris. The RBD was modified with addition of flexible N- and C-terminal amino acid extensions aimed to modulate the protein/protein interactions and facilitate protein purification. Fermentation with yeast extract culture medium yielded 30-40 mg/L. After purification by immobilized metal ion affinity chromatography and hydrophobic interaction chromatography, the RBD protein was characterized by mass-spectrometry, circular dichroism, and binding affinity to angiotensin-converting enzyme 2 (ACE2) receptor. The recombinant protein shows high antigenicity with convalescent human sera and also with sera from individuals vaccinated with the Pfizer-BioNTech mRNA or Sputnik V adenoviral-based vaccines. The RBD protein stimulates IFN{gamma}, IL-2, IL-6, IL-4, and TNF in mice secreting splenocytes from PBMC and lung CD3+ enriched cells. Immunogenicity studies with 50 {micro}g of the recombinant RBD formulated with alum, induce high levels of binding antibodies in mice and non-human primates, assessed by ELISA plates covered with RBD protein expressed in HEK293T cells. The mouse sera inhibited the RBD binding to ACE2 receptor in an in-vitro test and show neutralization of SARS-CoV-2 infection of Vero E6 cells. These data suggest that the RBD recombinant protein expressed in yeast P. pastoris is suitable as a vaccine candidate against COVID-19. HighlightsO_LIThe RBD protein (C-RBD-H6 PP) is expressed with high purity in P. pastoris. C_LIO_LIPhysico-chemical characterization confirms the right folding of the protein. C_LIO_LIThe recombinant protein shows high antigenicity with sera from convalescents. C_LIO_LIThe sera from animals inhibit the RBD-ACE2 binding and neutralize the virus. C_LIO_LIThe C-RBD-H6 protein stimulates IFN{gamma}, IL-2, IL-6, IL-4, and TNF in mice. C_LI

In vivo cholinergic basal forebrain degeneration and cognition in Parkinson's disease: Imaging results from the COPPADIS study.

INTRODUCTION: We aimed to assess associations between multimodal neuroimaging measures of cholinergic basal forebrain (CBF) integrity and cognition in Parkinson's disease (PD) without dementia. METHODS: The study included a total of 180 non-demented PD patients and 45 healthy controls, who underwent structural MRI acquisitions and standardized neurocognitive assessment through the PD-Cognitive Rating Scale (PD-CRS) within the multicentric COPPADIS-2015 study. A subset of 73 patients also had Diffusion Tensor Imaging (DTI) acquisitions. Volumetric and microstructural (mean diffusivity, MD) indices of CBF degeneration were automatically extracted using a stereotactic CBF atlas. For comparison, we also assessed multimodal indices of hippocampal degeneration. Associations between imaging measures and cognitive performance were assessed using linear models. RESULTS: Compared to controls, CBF volume was not significantly reduced in PD patients as a group. However, across PD patients lower CBF volume was significantly associated with lower global cognition (PD-CRStotal: r = 0.37, p < 0.001), and this association remained significant after controlling for several potential confounding variables (p = 0.004). Analysis of individual item scores showed that this association spanned executive and memory domains. No analogue cognition associations were observed for CBF MD. In covariate-controlled models, hippocampal volume was not associated with cognition in PD, but there was a significant association for hippocampal MD (p = 0.02). CONCLUSIONS: Early cognitive deficits in PD without dementia are more closely related to structural MRI measures of CBF degeneration than hippocampal degeneration. In our multicentric imaging acquisitions, DTI-based diffusion measures in the CBF were inferior to standard volumetric assessments for capturing cognition-relevant changes in non-demented PD.

Long term positivity of SARS-CoV-2 total immunoglobulins in convalescent plasma and blood donors

BackgroundOne of the most questioned issues about SARS-CoV2 immunity is how long does it last. Whether lasting differences exist between infection and vaccination boosted immunity is yet to be known. The answer to this question will determine key issues such as the reliability of individual and herd immunity or the need of sanitary restrictions or periodical revaccination. The aim of this study was to determine how long total anti SARS-CoV2 antibodies due to past infection persist in peripheral blood and whether sex, age or haematological features can influence their lasting. Material and MethodsA total of 2432 donations SARS-CoV-2 from 662 repeat donors from April 2020 to February 2021 were analysed. Donors were 69.7% males and their average age was 46. An automated chemilumiscence immunoassay for total antibodies recognizing N protein of SARS-CoV-2 in human serum and plasma was performed. Results and discussionIn 97.6% donors with follow-up, anti SARS-CoV-2 protein N total antibodies remained positive up to 46 weeks after first positive determination. Blood group was not related to antibody waning. Lower lymphocyte counts and higher neutrophils and as well higher seric IgA would help predict future negativization of antibodies. The vast majority of donors keep their total immunoglobulins anti SARS-CoV-2 positive for longer than 10 months. Ageing might have a protective effect against antibody waning but, given the small number of cases that become negative, more studies, or larger cohorts would be needed to confirm these facts.

Sustained seroprevalence of anti SARS-CoV-2 total immunoglobulins in asymptomatic blood donors

Background and objectivesCOVID-19 can either cause death or go unnoticed but antibodies will remain protecting us of SARS-CoV-2 reinfection for an uncertain time and to an uncertain extent. Our aim was to describe seroprevalence evolution from summer 2019 to autumn 2020 in Spain and to describe its relationship with age, blood group and haematological parameters. Materials and methodsSera and plasma from historical donation archives excluding convalescent were randomized and a total of 12,313 donations tested by a Chemiluminiscent analysis for anti SARS-CoV-2 N protein total immunoglobulins. Blood donors were 60.9% males, average age 46+/-13. Sex, age, blood group, blood cell counts and percentages and immunoglobulin concentrations were extracted from electronic recordings. ResultsA seroprevalence of 6.7% in blood donors was found by the end of the first wave. No differences by sex, age or blood group were found regarding antibodies. Leukocyte count (p=0.026), haematocrit (p<0.001) and haemoglobin (p<0.001) were lower in positive donations than in negative ones. Sex differences were present in neutrophils, leukocytes, haemoglobin and haematocrit as related to SARS-CoV-2 antibodies. ConclusionsSeroprevalence due to asymptomatic cases would resemble that of global population. Sex and age would not affect COVID-19 susceptibility but its severity. Gender differences related to COVID-19 in leukocytes, haemoglobin and haematocrit would be present in asymptomatic individuals. Further studies are needed to confirm these gender differences as they can help better understand the immune response to COVID-19, its pathogenesis and prognosis.

In-solution buffer-free digestion for the analysis of SARS-CoV-2 RBD proteins allows a full sequence coverage and detection of post-translational modifications in a single ESI-MS spectrum

Subunit vaccines based on the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2, are among the most promising strategies to fight the COVID-19 pandemic. The detailed characterization of the protein primary structure by mass spectrometry (MS) is mandatory, as described in ICHQ6B guidelines. In this work, several recombinant RBD proteins produced in five expression systems were characterized using a non-conventional protocol known as in-solution buffer-free digestion (BFD). In a single ESI-MS spectrum, BFD allowed very high sequence coverage ([≥] 99 %) and the detection of highly hydrophilic regions, including very short and hydrophilic peptides (2-8 amino acids), the His6-tagged C-terminal peptide carrying several post-translational modifications at Cys538 such as cysteinylation, glutathionylation, cyanilation, among others. The analysis using the conventional digestion protocol allowed lower sequence coverage (80-90 %) and did not detect peptides carrying some of the above-mentioned post-translational modifications. The two C-terminal peptides of a dimer [RBD(319-541)-(His)6]2 linked by an intermolecular disulfide bond (Cys538-Cys538) with twelve histidine residues were only detected by BFD. This protocol allows the detection of the four disulfide bonds present in the native RBD and the low-abundance scrambling variants, free cysteine residues, O-glycoforms and incomplete processing of the N-terminal end, if present. Artifacts that might be generated by the in-solution BFD protocol were also characterized. BFD can be easily implemented and we foresee that it can be also helpful to the characterization of mutated RBD.

Le corps d'adolescent au Rorschach / The body of the adolescent at the Rorschach / O corpo do adolescente no Rorschach / El cuerpo del adolescente en el Rorschach

Sur la base d'un cadre théorique qui est inscrit dans les théories de la relation d'objet, dans les processus de transformation et de co-construction, l'autour a l'intention d'observer et de décrire les processus de transformation. Les nouveaux organisateurs psychiques «techne¼ et «champ¼ compris comme une relation contenant-contenu permettent la compréhension des transformations psychiques de l'adolescence. Le Rorschach permet l'accès à la vie mentale et relationnelle du sujet, compris comme une relation de communication, de liaison et de transformation entre le sujet et l'objet. L'analyse du protocole du Rorschach, sera réalisée avec les deux organisateurs, par lesquelles nous cherchons à clarifier les relations établies entre le corps et le psychisme. L'extension à d'autres modèles théoriques de la psychanalyse et l'augmentation des possibilités de la méthodologie Rorschach nous permettent une meilleure compréhension des transformations du corps, dotant la pratique clinique de plus de possibilités pour l'intervention psychothérapeutique moins inscrite dans la logique classique de la psychopathologie.

Com base em quadro teórico inscrito nas teorias da relação de objeto, nos processos de transformação e de coconstrução, a autora tem por intenção observar e descrever os processos de transformação. Os novos organizadores psíquicos "techne" e "campo" inscritos em uma relação do tipo continente-conteúdo permitem a compreensão das transformações psíquicas dos adolescentes. O Rorschach permite o acesso à via mental e relacional do sujeito, compreendida como uma relação de comunicação, de ligação e de transformação entre o sujeito e o objeto. A análise do protocolo de Rorschach será realizada com base nos dois organizadores, através dos quais será possível esclarecer as relações que se estabelecem entre o corpo e o psiquismo. A extensão a outros modelos teóricos da psicanálise e o aumento das possibilidades da metodologia Rorschach permitem uma melhor compreensão das transformações do corpo, dotando a prática clínica de mais possibilidade ao nível da intervenção psicoterapêutica, a qual será menos inscrita na lógica clássica da psicopatologia.

Using a theoretical framework based on the theories of object relation, on the processes of transformation and co-construction, the author aims at observing and describing transformation processes. The new psychic organizers "techne" and "field", which form a container-contained relationship, help understand the psychic transformations of adolescents. The Rorschach test allows to access the mental and relational life of the subject, understood as a relationship of communication, connection and transformation between the subject and the object. The Rorschach protocol will be analyzed based on the two organizers, which helped clarify the relations between the body and the psyche. The extension to other theoretical psychoanalytical models and the increase in possibilities of the Rorschach methodology allow a better understanding of body transformations, providing clinical practice with increased possibilities for psychotherapeutic interventions that are less situated in the classical logic of psychopathology.

Teniendo como base un cuadro teórico con mención en las teorías de la relación de objeto, en los procesos de transformación y de coconstrucción, la autora tiene la intención de observar y describir los procesos de transformación. Los nuevos organizadores psíquicos "téchne" y "campo" mencionados en una relación tipo continente-contenido permiten la comprensión de las transformaciones psíquicas de los adolescentes. El Rorschach permite el acceso a la vía mental y relacional del sujeto, comprendida como una relación de comunicación, conexión y transformación entre el sujeto y el objeto. El análisis del protocolo de Rorschach se hace con base en los dos organizadores, a través de los cuales será posible aclarar las relaciones que se establecen entre el cuerpo y el psiquismo. La ampliación a otros modelos teóricos del psicoanálisis aumenta las posibilidades de la metodología Rorschach, facilitando una mejor comprensión de las transformaciones del cuerpo, dotando a la práctica clínica de más posibilidades a nivel de intervención psicoterapéutica, menos limitada a la lógica clásica de la psicopatología.

Auf der Grundlage eines theoretischen Rahmens, in Bezug auf die Theorien der Objektbeziehung und auf den Veränderungs- und Co-Konstruktionsprozessen, beabsichtigt die Verfasserin, die Veränderungsprozesse zu beobachten und zu beschreiben. Die neuen psychischen Konzepte „Techne" und „Feld", die auf einer Container-Contained (Behälter-Inhalt) Beziehung basieren, ermöglichen das Verständnis der psychischen Veränderung von Heranwachsenden. Der Rorschach Test ermöglicht den Zugang zu einer Person über einen geistigen und beziehungsmäßigen Weg, welcher als eine Beziehung von Kommunikation, Verbindung und Veränderung zwischen der Person und dem Objekt verstanden wird. Die Analyse eines Rorschachprotokolls wird auf der Grundlage beider Konzepte durchgeführt, über die es möglich sein wird, die Beziehungen zu verdeutlichen, die zwischen Körper und Psyche entstehen. Die Erweiterung auf andere theoretische Modelle der Psychoanalyse und die Erhöhung der Möglichkeiten der Rorschach-Methode verbessert das Verständnis der Veränderungen des Körpers und verleiht der klinischen Praxis mehr Möglichkeiten auf Ebene der psychotherapeutischen Intervention, welche dann weniger auf die klassische Logik der Psychopathologie basiert.

Effects of different protein sources on fermentation metabolites and nutrient digestibility of brachycephalic dogs.

Benefits to microbial fermentation in the colon and as a consequence less flatulence can be promoted for the health of adult dogs according to the amount and protein source. The present study evaluated different protein sources in dry food for brachycephalic dogs regarding microbial fermentation and nutrient digestibility. Four dry dog foods with similar protein content were formulated for adult maintenance: poultry meal (PM) diet; wheat gluten (WG) diet; PM + WG diet; and PM + WG + hydrolysed protein (HP) diet. Eight French bulldog adult dogs were arranged in a 4 × 4 Latin square design during the 28 d trial. Fresh faeces were collected for assessment of nutrient digestibility and analyses of faecal pH, SCFA, biogenic amines, ammonia and lactate. Means were compared by the PROC MIXED procedure of SAS and by Tukey's test, considering P ≤ 0·05. The animals fed the WG and PM + WG diets showed higher digestibility for DM (P < 0·05), organic matter (P < 0·05), crude protein (P < 0·001) and lower faeces production (P < 0·02) than the PM and PM + WG + HP diets. Feeding diet PM + WG + HP resulted in lower faecal score and pH (P < 0·05) compared with other diets. Concentrations of fermentation metabolites were not statistically significantly different among diets. In conclusion, WG alone or in combination with PM improved protein and DM digestibility. Fermentation products were not affected by protein source.

Reverse Iontophoretic Extraction of Metabolites from Living Plants and their Identification by Ion-chromatography Coupled to High Resolution Mass Spectrometry.

INTRODUCTION: The identification and characterisation of cellular metabolites has now become an important strategy to obtain insight into functional plant biology. However, the extraction of metabolites for identification and analysis is challenging and, at the present time, usually requires destruction of the plant. OBJECTIVE: To detect different plant metabolites in living plants with no pre-treatment using the combination of iontophoresis and ion-chromatography with mass spectrometry detection. METHODOLOGY: In this work, the simple and non-destructive method of reverse iontophoresis has been used to extract in situ multiple plant metabolites from intact Ocimum basilicum leaves. Subsequently, the analysis of these metabolites has been performed with ion chromatography coupled directly to high resolution mass spectrometric detection (IC-MS). RESULTS: The application of reverse iontophoresis to living plant samples has avoided the need for complex pre-treatments. With this approach, no less than 24 compounds, including organic acids and sugars as well as adenosine triphosphate (ATP) were successfully detected. CONCLUSION: The research demonstrates that it is feasible to monitor, therefore, a number of important plant metabolites using a simple, relatively fast and non-destructive approach. Copyright © 2016 John Wiley & Sons, Ltd.

Effect of the addition of papain enzyme on digestive parameters and palatability of extruded diets for dogs / Efeito da adição da enzima papaína em rações extrusadas para cães sobre parâmetros digestivos e palatabilidade

Common protein sources used in the manufacturing of diets for dogs are derived from by-products, which may have reduced digestibility depending on the source. This study evaluated the effect of the addition of a protease, the papain enzyme, as a supplement to extruded diets on palatability, nutrient digestibility, and fecal production and quality of dogs. A diet was formulated with poultry by-product meal, meat and bone meal, and feather meal as protein sources. This formula was divided into three isonutrient diets: one negative control (NC), without enzymes; treatment one (EZ1) with addition of 855.000UI of papain per kilogram of diet, and treatment two (EZ2) with addition of 2.280.000UI of papain per kilogram of diet, both added before extrusion. The experiment followed a randomized block design, with two blocks of nine animals (three animals per treatment in each block), 18 dogs in total, and six replicates per treatment. Data were submitted to analysis of variance and the means of three treatments were compared by polynomial contrasts (P <0.05). No differences in the coefficients of total tract apparent digestibility of nutrients nor changes in palatability, pH, and fecal production among treatments were found with the addition of different doses of enzyme to the diets (P > 0.05). The fecal score was reduced with increased addition of enzyme (P < 0.05).(AU)

As fontes comuns de proteína utilizadas na fabricação de rações para cães são oriundas de coprodutos, os quais podem apresentar digestibilidade reduzida de acordo com a fonte. Este estudo avaliou os efeitos da adição da enzima papaína em dietas secas e extrusadas na palatabilidade, digestibilidade dos nutrientes, qualidade e produção fecal de cães adultos. Uma dieta foi formulada contendo farinha de vísceras de frango, farinha de carne e ossos e farinha de penas hidrolisadas como fontes proteicas. Esta foi posteriormente dividida em três dietas isonutrientes: controle negativo (CN) sem adição da enzima; adição de 855.000 UI de papaína por quilograma de ração (EZ1); e adição de 2.280.000 UI de papaína por quiilograma de ração (EZ2), ambas adições feitas antes da extrusão. O experimento seguiu delineamento em blocos casualizados, com dois blocos de nove animais (três animais por tratamento em cada bloco), totalizando 18 cães, e seis repetições por tratamento. Os dados obtidos foram submetidos a análise de variância, com as médias dos três tratamentos comparadas por contrastes polinomiais (P < 0,05). Não foram verificadas diferenças nos coeficientes de digestibilidade aparente dos nutrientes ou mesmo alterações na palatabilidade, pH e produção de fezes entre os tratamentos com diferentes inclusões de enzima (P > 0,05). Apenas o escore fecal reduziu com o aumento da adição da enzima (P < 0,05).(AU)

Nutritional composition and evaluation of different methodologies for fat determination in wet feed for dogs and cats / Composição nutricional e avaliação de diferentes métodos de determinação de gordura em alimentos úmidos para cães e gatos

As a consequence of the increasing number of dog and cat owners, the pet food industry is expanding the range of pet food products in the market. In order to obtain more necessary information about the wet food segment for dogs and cats, the aim of this study was to determine the nutritional composition, to evaluate the information declared on the labels, and to compare the composition with the FEDIAF recommendations for protein and fat. Furthermore, three different methodologies of fat analysis were compared: crude fat (CFa), crude fat after acid hydrolysis (CFAH), and fat content obtained with Ankom XT15 (ANKOM) to determine the most adequate method for fat determination in wet foods. Twenty-five wet food products were evaluated, 13 wet foods for dogs and 12 for cats. Centesimal composition analyses obtained in this study were compared with guaranteed analysis declared on the label and with FEDIAF minimum recommended requirements for each species. The results of the nutritional composition and the values described on the label and the evaluation of the three fat determination methods were compared using the mixed model test with repeated measurements in the same samples, respectively (p < 0.05) in the SAS program, evaluation of protein adequacy and fat content were analyzed by mathematical calculations of difference and proportion. No difference was observed between nutritional composition of wet foods and the values declared on the labels for the majority of the diets analyzed, and there was a predominance of products that exceeded FEDIAF minimum recommendations of protein and fat for both species. No difference was observed between the three methods of fat content evaluation (p = 0.68). It was concluded that wet foods evaluated in this study match the label information and FEDIAF nutrient requirement recommendations, considering recommended calorie intake. All three fat determination methodologies evaluated were similar, justifying the choice of the easiest or cheapest method.(AU)

Devido ao aumento do número de cães e gatos domiciliados, a indústria de alimentos para animais de estimação tem expandido a gama de produtos existentes no mercado de pet food. Para obter informações mais relevantes sobre o segmento de alimentos úmidos para cães e gatos, este trabalho determinou a composição nutricional, avaliou as informações declaradas nos rótulos e comparou a composição com as recomendações da Fediaf de proteína e gordura. Também foram comparadas três metodologias diferentes de análise de gordura: extrato etéreo (CFa), extrato etéreo após hidrólise ácida (CFAH) e teor de gordura obtido no analisador Ankom XT15 (ANKOM) para determinar o método mais adequado de avaliação de gordura em alimentos úmidos. Foram avaliadas 25 marcas de alimentos úmidos, 13 para cães e 12 para gatos. As análises de composição centesimal obtidas neste estudo foram comparadas com a informação nutricional declarada nos rótulos e com as necessidades mínimas recomendadas pela Fediaf para cada espécie. Os resultados da composição nutricional, os valores descritos no rótulo e a avaliação dos três métodos para determinação da gordura foram comparados com o emprego do teste t e modelo misto com medidas repetidas nas mesmas amostras, respectivamente (p < 0,05) no programa SAS. Já a avaliação da adequação nutricional de proteína e do teor de gordura foram analisados por cálculos matemáticos de diferença e proporção. Para a maioria dos alimentos avaliados não foi observada diferença entre a composição nutricional dos alimentos úmidos e os valores declarados em rótulo, e houve predominância de produtos que excederam as recomendações mínimas de proteína e gordura da Fediaf para ambas as espécies. Quanto às metodologias de extração de gordura, não foi observada diferença entre os três métodos avaliados (p = 0,68). Concluiu-se que os alimentos úmidos avaliados atendem às informações declaradas pelos fabricantes e também às recomendações nutricionais da Fediaf com base na ingestão energética recomendada. Em relação às metodologias avaliadas para determinação de gordura nestes alimentos, a similaridade entre tais resultados justifica o uso da técnica de maior facilidade ou de menor custo.(AU)

Description and consequences of prescribing off-label antiretrovirals in the Madrid Cohort of HIV-infected children over a quarter of a century (1988-2012).

BACKGROUND: Licensing data for paediatric dosing is often sparse and subsequent studies may result in changes to recommended doses. We measured the extent and consequences of off-label antiretroviral (ARV) use in an HIV-infected paediatric cohort. METHODS: In this multicentre cohort study involving 318 HIV-infected children and adolescents from the Madrid Cohort, all off-label prescriptions from March 1988 to March 2012 were recorded from the clinical records. The reasons for prescribing ARV off-label, the side effects and the consequences of incorrect dosing of ARVs are discussed. RESULTS: Among the 318 patients of the cohort, 221 (69%) received off-label ARVs according to EMA licensing at the time of prescription, representing 23% (540) of the 2,353 prescribed ARVs. The main reason for starting an off-label drug was treatment failure. Adverse events led to treatment discontinuation in 12% of the prescriptions. Problems taking the drug led to withdrawal in 5%, more likely when formulation was not suitable for age (P<0.05). Up to 10% were overdosed and 10% underdosed, defined as 25% above or below the current recommended dose, respectively. Treatment failure occurred significantly more frequently among underdosed compared to overdosed patients (50% versus 26%; P<0.05). CONCLUSIONS: Off-label use of ARVs was common in our HIV-1 paediatric patients. Adverse events were common but rarely led to withdrawal. Suitable formulation is important in younger children. Pharmacokinetic studies are needed as frequent incorrect dosing may occur when prescribing off-label and underdosing may lead to treatment failure.

Plasma and urine metabolic fingerprinting of type 1 diabetic children.

Type 1 diabetes mellitus is one of the most common chronic disorders of childhood. The metabolic control is lost due to the lack of insulin, which is the main treatment for the disease. Nevertheless, long-term complications appear even under good glycemic control. Metabolomics, an emerging strategy, can help in diagnosis, prognosis, and monitoring of metabolic disorders. The objective of the present study was to investigate the alterations in plasma (by LC-MS) and urine (CE-MS) of type 1 diabetic children that were under insulin treatment and good glycemic control. Even without remarkable biochemical differences between the two groups (diabetic and control) except for glucose level and glycosilated hemoglobin, metabolomic tools were able to capture subtle metabolic differences. The main changes in plasma were associated to lipidic metabolism (nonesterified fatty acids, lysophospholipids, and other derivatives of fatty acids), and some markers of the differential activity of the gut microflora were also found (bile acids, p-cresol sulfate). In urine, changes associated to protein and amino acid metabolism were found (amino acids, their metabolites and derivatives), and among them one advanced glycation end product (carboxyethylarginine) and one early glycation end product (fructosamine) were excreted in higher proportion in the diabetic group.