Entrenamiento de la marcha con la cinta rodante antigravitatoria en pacientes con Parkinson / Gait training with the anti-gravity treadmill in Parkinson's patients / Treinamento de marcha com fita rolante antigravitatoria em pacientes com Parkinson

La enfermedad de Parkinson (EP) es una condición neurodegenerativa caracterizada por alteraciones motoras que afectan principalmente el desarrollo de la marcha, produciéndose generalmente el fenómeno del congelamiento de la marcha con la posibilidad del riesgo de caída. Objetivo: Analizar los beneficios del entrenamiento de la marcha con la cinta rodante antigravitatoria en pacientes con EP. Materiales y métodos: Es un estudio pre-experimental con un solo grupo de intervención. Se llevó a cabo el entrenamiento de la marcha a través de una cinta rodante antigravitatoria (AlterG) durante un mes dividido en 2 sesiones por semana. Los parámetros de la marcha arrojados por el equipo AlterG fueron la descarga de peso, cadencia, tiempo de apoyo y longitud del paso; además se valoró el congelamiento de la marcha con el cuestionario (FOGQ) y el riesgo de caída con el test Timed Up and Go (TUG). Resultados: En los parámetros de la marcha y en el FOGQ se encontró diferencias significativas (p<0,05) entre los valores de pre y post intervención, exceptuando a la variable cadencia. El riesgo de caída disminuyó 4,6 y 4,3 segundos promedio en el test TUG en hombres y mujeres respectivamente. Conclusión: El entrenamiento de la marcha en la cinta rodante antigravitatoria mejora las condiciones de la marcha y reduce el riesgo de caídas en pacientes con EP.

Parkinson's disease (PD) is a neurodegenerative condition characterized by motor alterations that mainly affect the development of gait, generally producing the phenomenon of freezing of gait with the possibility of risk of falling. Objective: To analyze the benefits of gait training with the antigravity treadmill in patients with PD. Materials and methods: It is a pre-experimental study with a single intervention group. Gait training was carried out using an antigravity treadmill (AlterG) for one month divided into 2 sessions per week. The gait parameters returned by the AlterG team were weight unloading, cadence, support time and step length; Furthermore, freezing of gait was assessed with the questionnaire (FOGQ) and the risk of falling with the Timed Up and Go test (TUG). Results: In the gait parameters and in the FOGQ, significant differences (p<0.05) were found between the pre- and post-intervention values, except for the cadence variable. The risk of falling decreased by 4.6 and 4.3 seconds on average in the TUG test in men and women respectively. Conclusion: Antigravity treadmill gait training improves walking conditions and reduces the risk of falls in PD patients.

A doença de Parkinson (DP) é uma condição neurodegenerativa caracterizada por alterações motoras que afetam principalmente o desenvolvimento da marcha, geralmente produzindo o fenômeno de congelamento da marcha com possibilidade de risco de queda. Objetivo: Analisar os benefícios do treino de marcha com esteira antigravitacional em pacientes com DP. Materiais e métodos: Trata-se de um estudo pré-experimental com um único grupo de intervenção. O treinamento de marcha foi realizado em esteira antigravitacional (AlterG) durante um mês dividido em 2 sessões semanais. Os parâmetros de marcha retornados pela equipe AlterG foram descarga de peso, cadência, tempo de apoio e comprimento do passo; Além disso, o congelamento da marcha foi avaliado com o questionário (FOGQ) e o risco de queda com o teste Timed Up and Go (TUG). Resultados: Nos parâmetros da marcha e no FOGQ foram encontradas diferenças significativas (p<0,05) entre os valores pré e pós-intervenção, exceto na variável cadência. O risco de queda diminuiu em média 4,6 e 4,3 segundos no teste TUG em homens e mulheres respectivamente. Conclusão: O treino de marcha em esteira antigravitacional melhora as condições de marcha e reduz o risco de quedas em pacientes com DP.

Application of Cut-and-Sew Technique in Thoracoscopic Minimally Invasive Mitral Valve Surgery and Concomitant Maze Procedure

ABSTRACT Introduction: Atrial fibrillation is one of the common complications of mitral valve disease. Currently, in the absence of freezing equipment, it's still impossible to fully conduct a minimally invasive Cox-maze IV procedure to treat atrial fibrillation. Methods: We analyzed the clinical data of 28 patients who underwent thoracoscopic minimally invasive mitral valve full maze surgery in our hospital from October 2021 to September 2022; 13 patients were male and 15 were female, three suffered from paroxysmal atrial fibrillation, and 25 suffered from permanent atrial fibrillation; average age was 61.88±8.30 years, and mean preoperative left atrial diameter was 47.12±8.34 mm. Isolation of left atrial posterior wall (box lesion) was completed in all patients by cut-and-sew technique and bipolar clamp ablation. Results: For these subjects, the median cardiopulmonary bypass time was 169 (109.75-202.75) minutes, aortic cross-clamping time was 106 (77.75-125.50) minutes, and ventilator assistance time was 6.5 (0-10) hours. Among them, eight subjects had the endotracheal tubes removed immediately after surgical operation. Three subjects were in the blanking period; two subjects still had atrial fibrillation at three months after operation, one of whom resumed sinus rhythm after electrical cardioversion therapy; and all the remaining 23 subjects had sinus rhythm. Conclusion: The minimally invasive cut-and-sew technique for electrical isolation of left pulmonary veins can improve sinus conversion rate of patients suffering from both mitral valve disease and atrial fibrillation. In selected subjects, it is safe and has good results in the short-term postoperative period.

Clinical assessment of upper limb impairments and functional capacity in Parkinson's disease: a systematic review / Avaliação clínica de comprometimentos de membros superiores e capacidade funcional na doença de Parkinson: uma revisão sistemática

Abstract Background Parkinson's disease (PD) may progressively reduce the upper limb's functionality. Currently, there is no standardized upper limb functional capacity assessment in PD in the rehabilitation field. Objective To identify specific outcome measurements to assess upper limbs in PD and access functional capacity. Methods We systematically reviewed and analyzed the literature in English published from August/2012 to August/2022 according to PRISMA. The following keywords were used in our search: "upper limbs" OR "upper extremity" and "Parkinson's disease." Two researchers searched independently, including studies accordingly to our inclusion and exclusion criteria. Registered at PROSPERO CRD42021254486. Results We found 797 studies, and 50 were included in this review (n = 2.239 participants in H&Y stage 1-4). The most common upper limbs outcome measures found in the studies were: (i) UPDRS-III and MDS-UPDRS to assess the severity and progression of PD motor symptoms (tremor, bradykinesia, and rigidity) (ii) Nine Hole Peg Test and Purdue Pegboard Test to assess manual dexterity; (iii) Spiral test and Funnel test to provoke and assess freezing of upper limbs; (iv) Technology assessment such as wearables sensors, apps, and other device were also found. Conclusion We found evidence to support upper limb impairments assessments in PD. However, there is still a large shortage of specific tests to assess the functional capacity of the upper limbs. The upper limbs' functional capacity is insufficiently investigated during the clinical and rehabilitation examination due to a lack of specific outcome measures to assess functionality.

Resumo Antecedentes A doença de Parkinson (DP) reduz progressivamente a funcionalidade do membro superior. Não existe uma avaliação padronizada da capacidade funcional do membro superior na DP na área da reabilitação. Objetivo Identificar medidas de resultados específicos para avaliar membros superiores na DP e avaliar capacidade funcional. Métodos Revisamos e analisamos sistematicamente a literatura publicada de agosto/2012 a agosto/2022 de acordo com PRISMA. Usamos as seguintes palavras-chave "membros superiores" OU "extremidade superior" e "doença de Parkinson." Dois pesquisadores fizeram a busca de forma independente, incluindo estudos de acordo com os critérios de inclusão e exclusão. Registro PROSPERO CRD42021254486. Resultados Encontramos 797 estudos, 50 foram incluídos no estudo(n = 2.239 participantes no estágio 1-4 de H&Y). As medidas de resultados de membros superiores mais comuns encontradas foram: (i) UPDRS-III e MDS-UPDRS, para avaliar a gravidade e a progressão dos sintomas motores da DP (tremor, bradicinesia, e rigidez); (ii) Nine Hole Peg Test e Purdue Pegboard Test para avaliar a destreza manual; (iii) Teste da Espiral e Teste do Funil para provocar e avaliar o congelamento de membros superiores; (iv) Avaliação de tecnologia, como sensores vestíveis, aplicativos e outros dispositivos também foram encontrados. Conclusão Encontramos evidências para dar suporte para as avaliações de deficiências de membros superiores na DP. No entanto, ainda há grande escassez de testes específicos para avaliar a capacidade funcional dos membros superiores. A capacidade funcional dos membros superior é insuficientemente investigada durante o exame clínico e de reabilitação devido à falta de medidas de resultados específicos para avaliar a funcionalidade.

Dimethyl acetamide and dimethyl sulfoxide associated at glucose and egg yolk for cryopreservation of Pseudoplatystoma corruscans semen

This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)

Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)

Effect of frozen storage on preservation of a silicone-based test food material

Aim: This study aimed to evaluate the effect of frozen storage on the physical properties of a silicone-based test food material, highly used to evaluate the masticatory performance in research settings. Methods: A total of 1,666 silicone cubes of Optosil Comfort® with 5.6-mm edges were shaped and stored at -18°C. The cubes were subsequently tested for flexural strength (maximum force, displacement, stress, and strain) before breaking (n = 136), changes in weight and size (n = 170), and masticatory performance (n = 1360) at eight timepoints: immediately after cube preparation (baseline, no freezing), and 1, 2, 3 and 4 weeks, and 2, 4 and 6 months after frozen storage. The cubes were thawed 8 h before each assessment. Results: The maximum force, stress, maximum displacement, and deformation values for the cubes were not affected by freezing (P > 0.05). At all of the time points, the cubes exhibited similar weight (P = 0.366) and size (identical values). The masticatory performance for the cubes also showed no differences from baseline through 6 months (P = 0.061). Conclusion: Freezing Optosil Comfort® silicone cubes did not alter the physical and mechanical properties of the material, being suitable to optimize the assessment of masticatory parameters for research purposes

Insights on sperm assays and cryopreservation in six Neotropical pit vipers

Breeding snakes in captivity has become more and more relevant due not only to the growing interest on their venoms but also to the increasing number of endangered species worldwide. Unfortunately, studies on the formation of germplasm banks for these reptiles do not follow the same pace, and literature on sperm cryopreservation remains in its infancy when compared to other taxa. Herein, we first validated a sperm-egg binding assay (using chicken egg perivitelline membrane – EPM) and some nonfluorescent staining techniques for semen analysis of two pit viper genera (Bothrops and Crotalus), and then we investigated the protective effects of dimethylacetamide (DMA), dimethylformamide (DMF), and dimethylsulfoxide (DMSO) at different concentrations (3, 6 and 12%) throughout the freezing process in five species of lancehead and one of rattlesnake. Our validation process showed high correlations among sperm functional tests (including sperm-binding to EPM) and motion parameters. A total of 166 fresh ejaculates were acquired from 233 collection attempts, and 63.9% of these samples exhibited minimal motility for freezing (≥20%). During cryopreservation we observed that post-thaw motility and quality was improved by higher levels of cryoprotectants (CPA), regardless the CPA type. Lower concentrations of CPA were less harmful to sperm motility and progressive motility following the equilibrium phase, but were ineffective in protecting these cells from the freeze-thaw cycle. Likewise, higher CPA concentrations increased post-thaw integrity of the acrosome and plasma membrane for most species, except for rattlesnakes in which only 12% DMSO produced better outcomes.

Quedas na doença de Huntington / Falls in Huntington's Disease

Introdução: Quedas estão associadas com injúrias, medo de cair, diminuição da mobilidade funcional e piora na qualidade de vida. Entretanto, pouco se sabe sobre os fatores potencialmente contribuintes para ocorrência de quedas na Doença de Huntington (DH) assim como sobre as circunstâncias nas quais elas ocorrem. Objetivos: Investigar as características clínicas potencialmente associadas com o aumento do risco de quedas na DH, assim como as circunstâncias nas quais as quedas ocorrem e suas consequências. Métodos: Dados demográficos e clínicos foram obtidos através de entrevista, revisão do prontuário e exame físico. Instrumentos para avaliação de risco de quedas (TUG), sintomas motores (UHDRS, UPDRS), distúrbios da marcha (10MWT, FOG), distúrbios cognitivos (MEEM, FAB, FDT, Hayling,Ekman), alterações comportamentais (Irritability Scale, BIS-11, BDI-II, NPI-Q) e distúrbios do equilíbrio (BBS) foram aplicado em todos pacientes. Resultados: Foram obtidos dados de 40 pacientes e 24 (60%) apresentaram ≥ 2 quedas nos últimos 6 meses e foram considerados "caidores recorrentes". Idade, idade de início da doença e duração da doença (DCL) não diferiram entre os grupos. Em contrapartida, a dose de neurolépticos medida em equivalentes de Olanzapina (OE) foi maior no grupo dos "caidores recorrentes." Os pacientes "caidores recorrentes" também apresentaram pior performance no desempenho da UHDRS-TMS, UPDRS, BBS e mais coreia do que os "não-caidores". Houve também diferença estatística na comparação dos tempos de escolha e alternância do FDT indicando pior desempenho cognitivo do grupo dos "caidores recorrentes" no teste. "Caidores- recorrentes" também apresentaram mais comportamento motor aberrante do que os "não- caidores". Não houve diferença na comparação dos parâmetros espaço-temporais da marcha estudados, assim como no desempenho do TUG. Ambos os grupos apresentaram altos índices de medo de queda. Somente o modelo contento o BBS alcançou significância estatística na regressão logística. Cerca de 80% das quedas ocorreram dentro de casa, caminhar foi o ato mais comum durante as quedas. Ainda, 40% das quedas foram classificadas como intrínsecas. Conclusão: As quedas são frequentes na DH assim como o medo de cair. Nosso estudo sugere que o risco de quedas na doença de Huntington seja multifatorial. Altas doses de neurolépticos, coreia, sintomas cognitivos e comportamentais e particularmente distúrbios do equilíbrio contribuem para ocorrência de quedas na HD.

Introduction: Falls are associated with injuries, fear of falling, decreased functional mobility and worsening in quality of life. However, little is known about the potentially contributing factors to the occurrence of falls in Huntington's Disease (HD) as well as the circumstances in which they occur. Objectives: To investigate clinical features potentially associated with an increased risk of falls in HD, as well as the circumstances in which falls occur and their consequences.Methods: Demographic and clinical data was obtained through interviews, chart review and physical examination. Instruments for fall risk assessment (TUG), motor symptoms (UHDRS, UPDRS), gait disorders (10MWT, FOG), cognitive disorders (MMSE, FAB, FDT, Hayling, Ekman), behavioral changes (Irritability Scale, BIS- 11, BDI-II, NPI-Q) and balance disorders (BBS) were applied to all patients.Results: Data from 40 patients was obtained and 24 (60%) had ≥ 2 falls in the last 6 months and were considered "recurrent fallers". Age, age of disease onset and disease duration (DCL) did not differ between groups. In contrast, the dose of neuroleptics measured in olanzapine equivalents (EO) was higher in the "recurrent fallers" group. The "recurrent fallers" also showed worse performance in the execution of the UHDRS-TMS, UPDRS, BBS and more chorea than the "non-fallers". There was also a statistical difference in the comparison of times of choice and alternation of the FDT, indicating worse cognitive performance of the "recurrent fallers" group in the test. "Recurrent fallers" also showed more aberrant motor behavior than "non-fallers". There was no difference in the comparison of the spatio-temporal gait parameters studied, nor in the TUG performance. Both groups had high rates of fear of falling. Only the model containing the BBS reached statistical significance in the logistic regression. About 80% of falls occurred indoors, walking was the most common act during falls. In addition, 40% of falls were classified as intrinsic. Conclusion: Falls are frequent in HD as well as the fear of falling. Our study suggests that the risk of falls in Huntington's disease is multifactorial. High doses of neuroleptics, chorea, cognitive and behavioral symptoms and particularly balance disorders contribute to the occurrence of falls in HD.

Efectos de la necesidad cierre cognitivo en los estilos decisorios: un abordaje exploratorio

Resumen Los estilos decisorios se refieren a patrones cognitivos habituales que utilizan los sujetos en tareas decisorias cuando procesan la información. Recientes investigaciones han referido el rol que cumple la motivación epistémica de la Necesidad de Cierre Cognitivo en el procesamiento de la información. Esta influencia puede generar un procesamiento sesgado afectando la toma de decisiones. Esto se debe a que la NCC trae dos consecuencias en el procesamiento de la información. La primera es la Urgencia donde los individuos tienden a una búsqueda generalizada de la información generando mayores hipótesis hasta superar el umbral cognitivo. La segunda refiere a la Permanencia donde una vez superado el umbral, la información se cristaliza y se extrapola a otras áreas. El presente estudio exploratorio, tuvo como objetivo explorar la relación entre la Necesidad Cierre Cognitivo y los Estilos decisorios. Para ello, se tomó muestra de 378 individuos residentes en Ciudad de Buenos Aires y Gran Buenos Aires. Se utilizaron para evaluar las variables el Test Revisado de Necesidad de cierre cognitivo (TR NCC) y la escala de Estilos Decisorios (ED).Los resultados indican que si bien no existe una relación contundente entre ambas variables, este abordaje exploratorio debe seguir siendo explorado en futuras investigaciones.

Abstract Decisional styles refer to habitual cognitive patterns used by subjects in decision-making tasks when they process the information. Recent research has referred to the role played by the epistemic motivation of the Need for Cognitive Closure in information processing. This influence can generate a biased processing affecting decision making. This is because NCC brings two consequences in information processing. The first one is the Urgency where individuals tend to a generalized search for information generating more hypotheses until the cognitive threshold is exceeded. The second refers to Permanence, where once the threshold is surpassed, the information crystallizes and is extrapolated to other areas. The present exploratory study aimed to explore the relationship between the Need for Cognitive Closure and Decision Styles. For this purpose, a sample of 378 individuals residing in the City of Buenos Aires and Greater Buenos Aires was taken. The Revised Test of Need for Cognitive Closure (TR-NCC) and the Decision-Making Styles (DE) scale were used to assess the variables. The results indicate that although there is no strong relationship between the two variables, this exploratory approach should be further explored in future research.

Eficácia da criolipólise para redução de gordura subcutânea: uma revisão sistemática e metanálise / Effectiveness of cryolipolysis for subcutaneous fat reduction: a systematic review and meta-analysis

Introdução: a criolipólise produz um resfriamento seletivo e controlado e baseia-se no conceito de que tecidos ricos em lipídios são mais suscetíveis a lesões por frio, reduzindo gordura subcutânea. Objetivo: revisar a literatura para avaliar a eficácia da criolipólise na redução de gordura subcutânea. Métodos: revisão sistemática com metanálise de estudos publicados nas bases de dados EBSCOhost, LILACS e PUBMED. Resultados: apenas um estudo não apresentou redução significativa na gordura subcutânea quando comparado o grupo de intervenção ao grupo controle. Houve diferença entre os estudos em relação aos parâmetros. Conclusão: a criolipólise é uma ferramenta eficaz para redução de gordura localizada.

Introduction: Cryolipolysis produces selective, controlled cooling, and it's based on the concept that lipid-rich tissue is more susceptible to cold injury, reducing subcutaneous fat. Objective: To review the literature to assess the effectiveness of cryolipolysis in reducing subcutaneous fat. Methods: Systematic review with meta-analysis of studies published in the EBSCOhost, LILACS, and PUBMED databases. Results: Only one study did not present significant reduction in subcutaneous fat compared to the control group. There was a difference among the parameters in the studies. Conclusion: Cryolipolysis is an effective tool for localized fat reduction.

Study of the effects of freezing process on nutritional value of 17 fish species marketed in São Paulo State, Brazil / Estudo dos efeitos do processo de congelamento sobre o valor nutricional de 17 espécies de peixes comercializadas no Estado de São Paulo, Brasil

Freezing is an important strategy to keep fish quality and make the species available the whole year. Its effects on the nutritional value of 17 fish species were studied in samples of entire fish, fillets or pieces. One portion of homogenized flesh was analyzed just after purchase (fresh sample). The other portion was packed in polyethylene bag, sealed, quick frozen (-80°C), stored properly at -18°C and analyzed after 12 months (frozen sample). Moisture, ash and protein content were tested using Brazilian Supply, Livestock and Agriculture Ministry methodologies. Lipid content was analyzed through Bligh and Dyer method. Carbohydrate content and caloric value were calculated, using NIFEXT fraction and Atwater coefficient, respectively. When fresh and frozen samples were compared, moisture and ash content showed significant difference (p<0.05) for 17.65% and 11.77% species, respectively. Lipid and protein contents were the most affected parameters, as they were altered in 29.40% of the studied species (p<0.05), and therefore, highlighted the importance of the conservation technology used on nutritional quality of fishery products. Mullet (M. brasiliensis) and Atlantic salmon (S. salar) had their nutritional composition more affected by freezing process with five and four altered parameters, respectively, from the six studied. (AU)

O congelamento é estratégia importante para manter a qualidade do peixe e tornar inúmeras espécies disponíveis o ano todo. Seus efeitos sobre o valor nutricional de 17 espécies foram estudados em amostras de peixes inteiros, filés ou postas. A porção cárnea homogeneizada foi analisada logo após a aquisição (amostra fresca). Outra parte foi embalada em polietileno, selada, rapidamente congelada (-80°C) e analisada após 12 meses de armazenamento a -18°C (amostra congelada). O teor de umidade, cinzas e proteína foram testados com metodologias do Ministério de Agricultura, Pecuária e Abastecimento e teor de lipídios com método de Bligh e Dyer. Conteúdo de carboidrato e valor calórico foram calculados, utilizando fração NIFEXT e coeficiente de Atwater, respectivamente. Quando se comparou amostras frescas e congeladas, teor de umidade e cinzas evidenciaram diferenças significativas (p<0,05) para 17,65% e 11,77% das espécies, respectivamente. O teor de lipídios e de proteínas foram alterados em 29,40% das espécies estudadas (p<0,05), sendo os parâmetros mais afetados pelo congelamento e destacaram a importância da tecnologia de conservação utilizada sobre a qualidade nutricional do pescado. Tainha (M. brasiliensis) e salmão (S. salar) foram as mais afetadas pelo congelamento, com 5 e 4 parâmetros alterados, respectivamente, após estocagem sob congelamento. (AU)