A rational approach for the improvement of biomass production and lipid profile in cacao cell suspensions.

Cocoa butter (CB) is produced in the seeds of Theobroma cacao representing 50% of its dry weight. The lipid composition plays an important role in the physicochemical, rheological, and sensory properties of the CB, making this fat a valuable resource for the production of chocolates, cosmetics, and pharmaceuticals. In this paper, are described experimental strategies used for a rational improvement of biomass production and fatty acids in cacao cell suspension cultures. First, the lipid profile in four cacao varieties is characterized, and then, one variety is selected to induce cell suspensions using a direct method without previous establishment of a callus phase. To improve growth and total fat production in cell suspension cultures, modified DKW media and newly designed media culture, based on the mineral concentrations of cacao seeds (cacao biomass production, "CBP"), are analyzed and compared. In addition, the effect of acetate in the lipid profile of cell suspensions is evaluated. Ultrastructural histological analysis of lipid vesicles in cacao seeds and cell suspensions is also performed. The results will show that it is feasible to establish cacao suspensions without the calli step and increase the biomass production by selecting a suitable cacao variety and tissue and also applying a new culture media formulation. In addition, it is possible to synthesize fatty acids in cell cultures and modify the lipid profile adding a precursor of the novo biosynthesis of fatty acids such as the acetate. Transmission electronic microscopy examinations and differential interference contrast microscopy analysis will demonstrate that lipid vesicles are the main reserve substance in both cacao seeds and cell suspensions.

[Hospital Mortality in Parkinson's Disease: Retrospective Analysis in a Portuguese Tertiary Centre]. / Mortalidade Hospitalar na Doença de Parkinson: Análise Retrospetiva num Hospital Terciário Português.

INTRODUCTION: Parkinson's disease is associated with high hospital mortality. Male gender, late age at onset, higher disability and the coexistence of cognitive impairment or depression have been suggested to be risk factors of death. Pneumonia and cardiovascular diseases are the leading causes of death. OBJECTIVE: To characterize the mortality (causes of hospital admission and death) of Parkinson's disease patients in a tertiary hospital, as well as demographic and clinical characteristics. MATERIAL AND METHODS: Identification of hospital admissions of Parkinson's disease patients that resulted in inpatient death between 2008 and 2014. Retrospective review of medical files and inclusion of patients with disease clinically confirmed by a neurologist. Assessment of causes of death and demographic and clinical characteristics of patients. RESULTS: 1525 hospital admissions of Parkinson's disease patients were identified, of which 150 resulted in death. Of these, 52 patients met the inclusion criteria. Mean age at onset of Parkinson's disease symptoms was 66.8 years (± 8.7) and mean duration of disease was 12.5 years (± 7.9). Sixty-five percent of patients were in stages 4-5 of the Hoehn and Yahr scale. Thirty-three patients (63%) had dementia and eleven (21%) had depression. Infections were the leading cause of death (respiratory in 63% of cases). DISCUSSION: Similarly to literature, pneumonia was the leading cause of hospital death and most patients presented advanced disease stage and dementia. Contrarily to other studies, life expectancy was not reduced and cardiovascular diseases and trauma were not causes of death in our population. CONCLUSIONS: This is the first Portuguese mortality study in Parkinson's disease. Pneumonia is the leading cause of death. Advanced disease stage and dementia were common features in these patients.

Introdução: A doença de Parkinson está associada a elevada mortalidade hospitalar. O sexo masculino, o início tardio da doença, o grau de incapacidade e a coexistência de deterioração cognitiva ou de depressão têm sido apontados como fatores de risco. A pneumonia e as doenças cardiovasculares são as principais causas de morte. Objetivo: Explorar a mortalidade hospitalar (motivo de internamento e de óbito) dos doentes com doença de Parkinson num hospital terciário, assim como as características demográficas e clínicas. Material e Métodos: Identificação das admissões hospitalares dos doentes com o diagnóstico de doença de Parkinson entre 2008 e 2014 e seleção dos doentes falecidos. Revisão retrospetiva dos processos clínicos e inclusão dos doentes com doença confirmada clinicamente por um neurologista. Avaliação das causas de óbito, características demográficas e clínicas. Resultados: Identificámos 1 525 admissões hospitalares de doentes com diagnóstico de doença de Parkinson, das quais 150 resultaram em óbito. Destes, 52 cumpriam critérios de inclusão. A idade média do início dos sintomas de doença foi 66,8 anos (± 8,7) e a duração média da doença foi 12,5 anos (± 7,9). Sessenta e cinco por cento dos doentes encontravam-se no estádio 4-5 na escala de Hoehn e Yahr. Trinta e três doentes (63%) apresentavam demência e onze depressão. As infeções foram a principal causa de morte (respiratória em 63% dos casos). Discussão: Ã semelhança da literatura, a pneumonia foi a principal causa de morte hospitalar e a maioria dos doentes apresentava estádio avançado de doença e demência. Contrastando com outros estudos, não se verificou diminuição da esperança média de vida e as doenças cardiovasculares e o traumatismo não foram causas de morte na nossa população. Conclusões: Apresentamos o primeiro estudo português de mortalidade na doença de Parkinson. A pneumonia é a principal causa de morte hospitalar. O estádio avançado de doença e a demência foram características comuns nestes doentes.

Dyschromatopsia in Multiple Sclerosis Patients: A Marker of Subclinical Involvement?

BACKGROUND: In multiple sclerosis (MS), even in the absence of a clinical episode of optic neuritis (ON), the optic nerve and retinal nerve fiber layer (RNFL) may be damaged leading to dyschromatopsia. Subclinical dyschromatopsia has been described in MS associated with lower motor and cognitive performances. OBJECTIVES: To set the prevalence of dyschromatopsia in eyes of MS patients without a history of ON, to compare its prevalence in patients with and without ON history, and to explore the association between dyschromatopsia and disease duration, average peripapillary RNFL thickness, macular volume, and cognitive and motor performances. METHODS: An observational cross-sectional study was conducted at multiple medical centers. Data were collected after single neurological and ophthalmological evaluations. Dyschromatopsia was defined by the presence of at least 1 error using Hardy-Rand-Rittler plates. RESULTS: In our population of 125 patients, 79 patients (63.2%) never had ON and 35 (28.8%) had unilateral ON. The prevalence of dyschromatopsia in eyes of patients without ON was 25.7%. Patients with dyschromatopsia had a statistically significant lower RNFL thickness (P = 0.004 and P = 0.040, right and left eyes, respectively) and worse performance in symbol digit modalities test (P = 0.012). No differences were found in macular volume or motor function tasks. CONCLUSIONS: Dyschromatopsia occurs frequently in MS patients. It may be associated with a worse disease status and possibly serve as a marker for the detection of subclinical disease progression since it was detected even in the absence of ON. It correlated with thinner peripapillary RNFL thickness and inferior cognitive performance.

POLYPHENOLS DISTRIBUTION AND RESERVE SUBSTANCES ANALYSIS IN CACAO SOMATIC EMBRYOGENESIS / Análisis de distribución de polifenoles y sustancias de reserva en embriogénesis somática de cacao

In order to understand the causes of lack of regeneration in cacao somatic embryos, two cacao varieties with different responses to regeneration potential were described based on their capacity to store different compounds. It is well known that seed reserves play a central role in the regenerative capability of somatic embryos; thus, we followed histochemical changes and reserve fluctuations of proteins, polysaccharides and polyphenols during somatic embryogenesis (SE) in the two cacao varieties. The study showed that, in somatic embryos of the regenerating variety, polyphenols were localized mainly in the periphery of the embryo (epidermal cells) and proteins were the main storage substance in the embryo expression medium, while the non-regenerating variety had a high presence of polysaccharides with random distribution of polyphenols at the end of the embryo induction step.

Dos variedades de cacao con diferentes respuestas a la regeneración fueron descritas en función de su capacidad para almacenar diferentes compuestos, con el fin de aproximarse al entendimiento de las causas de la falta de regeneración en embriones somáticos de cacao. Es bien sabido que las reservas de semillas desempeñan un papel central en la capacidad de regeneración de embriones somáticos; por tanto, se realizó un seguimiento de cambios histoquímicos y fluctuaciones de reserva de proteínas, polisacáridos y polifenoles durante la embriogénesis somática (SE) en dos variedades de cacao. El estudio mostró que, en los embriones somáticos de la variedad regenerante, los polifenoles se localizaron principalmente en la periferia del embrión (células de la epidermis) y las proteínas fueron el componente principal de almacenamiento en el medio de expresión de embriones, mientras que la variedad no regenerante tenía una alta presencia de polisacáridos y una distribución aleatoria de los polifenoles en el final de la etapa de inducción de embriones.

Mielopatía espondilótica que simula una mielitis: claves diagnósticas mediante resonancia magnética / Spondylotic myelopathy mimicking myelitis: diagnostic clues by magnetic resonance imaging

Introducción. La mielopatía espondilótica es la causa más frecuente de mielopatía no traumática. Sus características radiológicas en ocasiones son indistinguibles de las de una mielopatía inflamatoria, y pueden ocasionar retrasos diagnósticos o llevar a un diagnóstico y tratamiento incorrectos. La descripción reciente de un patrón característico de realce de gadolinio puede ayudar a diferenciar la mielopatía espondilótica de la causa inflamatoria. Casos clínicos. Caso 1: varón de 38 años que consultó por historia de dos años de evolución de parestesias en las extremidades superiores, a las que se añadieron un año más tarde calambres en las extremidades derechas y acorchamiento en los dermatomas C5 y C6 en relación con los movimientos del cuello. Caso 2: varón de 44 años que consultó por historia de un año de evolución de alteración progresiva de la marcha con trastorno sensitivo en las manos y disfunción vesical reciente. En ambos casos, la resonancia magnética medular mostró en las secciones sagitales una hiperseñal cervical fusiforme en T2, asociada a una banda transversa de realce de gadolinio en forma de barra (pancakelike) justo por debajo de la zona de máxima estenosis, y un realce circunferencial o hemimedular en los cortes axiales. Conclusiones. Las características radiológicas de la mielopatía espondilótica pueden parecerse a las de origen inflamatorio. El reconocimiento de un realce transverso en barra inmediatamente por debajo del lugar de máxima compresión como signo radiológico típico de la mielopatía espondilótica es importante para reducir el riesgo de errores diagnósticos, y de utilidad para el manejo de estos pacientes (AU)

Introduction. Spondylotic myelopathy is the commonest cause of nontraumatic myelopathy. Radiological features of spondylotic myelopathy can often overlap with inflammatory myelopathies which may lead to a delayed or incorrect diagnosis and therapy. A distinctive gadolinium enhancement pattern recently described may help to differentiate spondylotic from inflammatory myelopathy. Case reports. Case 1: a 38-years-old man presented with a 2-year history of paresthesias in the upper extremities, and one year later cramps on the right limbs and numbness over right C5 and C6 dermatomes, related to movement of the neck. Case 2: a 44-year-old man presented with a 1-year history of progressive gait difficulties and sensory disturbance in the hands, and a recent onset of bladder dysfunction. In both cases, spinal cord MRI identified a longitudinal cervical T2-signal hyperintensity associated with a pancakelike transverse band of gadolinium enhancement just below the site of maximum spinal stenosis, and circumferential or hemicord enhancement on axial images. Conclusions. The radiological features of spondylotic myelopathy may resemble those of inflammatory origin. The recognition of a transverse pancakelike gadolinium enhancement immediately below the site of maximal compression as a typical radiological pattern of spondylotic myelopathy is important to reduce the risk of misdiagnosis and to help in the management of these patients (AU)

Spondylotic myelopathy mimicking myelitis: diagnostic clues by magnetic resonance imaging.

INTRODUCTION: Spondylotic myelopathy is the commonest cause of nontraumatic myelopathy. Radiological features of spondylotic myelopathy can often overlap with inflammatory myelopathies which may lead to a delayed or incorrect diagnosis and therapy. A distinctive gadolinium enhancement pattern recently described may help to differentiate spondylotic from inflammatory myelopathy. CASE REPORTS: Case 1: a 38-years-old man presented with a 2-year history of paresthesias in the upper extremities, and one year later cramps on the right limbs and numbness over right C5 and C6 dermatomes, related to movement of the neck. Case 2: a 44-year-old man presented with a 1-year history of progressive gait difficulties and sensory disturbance in the hands, and a recent onset of bladder dysfunction. In both cases, spinal cord MRI identified a longitudinal cervical T2-signal hyperintensity associated with a pancakelike transverse band of gadolinium enhancement just below the site of maximum spinal stenosis, and circumferential or hemicord enhancement on axial images. CONCLUSIONS: The radiological features of spondylotic myelopathy may resemble those of inflammatory origin. The recognition of a transverse pancakelike gadolinium enhancement immediately below the site of maximal compression as a typical radiological pattern of spondylotic myelopathy is important to reduce the risk of misdiagnosis and to help in the management of these patients.

TITLE: Mielopatia espondilotica que simula una mielitis: claves diagnosticas mediante resonancia magnetica.Introduccion. La mielopatia espondilotica es la causa mas frecuente de mielopatia no traumatica. Sus caracteristicas radiologicas en ocasiones son indistinguibles de las de una mielopatia inflamatoria, y pueden ocasionar retrasos diagnosticos o llevar a un diagnostico y tratamiento incorrectos. La descripcion reciente de un patron caracteristico de realce de gadolinio puede ayudar a diferenciar la mielopatia espondilotica de la causa inflamatoria. Casos clinicos. Caso 1: varon de 38 años que consulto por historia de dos años de evolucion de parestesias en las extremidades superiores, a las que se añadieron un año mas tarde calambres en las extremidades derechas y acorchamiento en los dermatomas C5 y C6 en relacion con los movimientos del cuello. Caso 2: varon de 44 años que consulto por historia de un año de evolucion de alteracion progresiva de la marcha con trastorno sensitivo en las manos y disfuncion vesical reciente. En ambos casos, la resonancia magnetica medular mostro en las secciones sagitales una hiperseñal cervical fusiforme en T2, asociada a una banda transversa de realce de gadolinio en forma de barra (pancakelike) justo por debajo de la zona de maxima estenosis, y un realce circunferencial o hemimedular en los cortes axiales. Conclusiones. Las caracteristicas radiologicas de la mielopatia espondilotica pueden parecerse a las de origen inflamatorio. El reconocimiento de un realce transverso en barra inmediatamente por debajo del lugar de maxima compresion como signo radiologico tipico de la mielopatia espondilotica es importante para reducir el riesgo de errores diagnosticos, y de utilidad para el manejo de estos pacientes.

Regeneración vía embriogénesis somática de una variedad colombiana élite de Theobroma cacao L / Regeneration through somatic embryogenesis of an elite colombian Theobroma cacao L variety

El objetivo de esta investigación fue evaluar dos protocolos de propagación vía embriogénesis somática a partir de explantes florales en dos clones élite BIOB e ICS95 de Theobroma cacao L. Se obtuvo un 50 y 32% de callo embriogénico en ICS95 y BIOB respectivamente con el protocolo de Fontanel et al. (2002), modificado después de un periodo de cultivo de tres meses. Los embriones pasaron por fases que se correspondieron con medios de cultivo diferenciales: Inducción, Formación, Maduración y Mantenimiento. Para la embriogénesis somática secundaria se obtuvo un 23% de embriones a partir de embriones somáticos primarios en un medio, conteniendo 1mg/L de 2,4,5 T (2,4,5 Triclorofenoxiacético). Se logró, además, desarrollar enraizamiento adventicio aplicando pulsos de IBA (Ácido Indol Butírico) a 0.5mg/L y 0.5g/L durante un minuto. Las plantas enraizadas se llevaron a una mezcla de tierra: arena (1:1) para su adaptación ex vitro, obteniéndose un 66% de plantas aclimatadas. Los estudios histológicos mostraron diferentes características típicas del desarrollo embriogénico. Este es el primer reporte en el que se logra de manera exitosa la conversión hasta plántula (68%) y la adaptación ex vitro de una variedad colombiana de cacao vía embriogénesis somática primaria y secundaria.

In this research we evaluate two protocols of propagation via somatic embryogenesis from floral explants using two elite clones BIOB and ICS95 of Theobroma cacao L. We obtained 50 and 32% of embryogenic callus on ICS95 and BIOB respectively with Fontanel et al., (2002) protocol modified after three months of culture. The embryos went through four phases; Induction, Formation, Maduration and Mantenimiento which corresponded each one with different media culture. For secondary somatic embryogenesis we obtained 23% of embryos from primary somatic embryos in a medium with 1mg/L of 2,4,5 T (2,4,5 Triclorofenoxiacetic). Also we obtained plants that developed new roots applying pulses with IBA (Indol Butiric Acid) 0.5mg/L and 0.5g/L for a minute. The developed plants were moved to a mix of potting soil and sand (1:1) for their ex vitro adaptation, getting 66% of acclimatized plants. The histological analysis showed the typical characteristics of the embryogenic development. This is the first report where it is achieved the successful conversion to plantlets (68%) and ex vitro adaptation of a colombian cocoa variety via primary and secondary embryogenesis.