The synergetic effect from the combination of different adsorption resins in batch and semi-continuous cultivations of S. Cerevisiae cell factories to produce acetylated Taxanes precursors of the anticancer drug Taxol.

In situ product recovery is an efficient way to intensify bioprocesses as it can perform adsorption of the desired natural products in the cultivation. However, it is common to use only one adsorbent (liquid or solid) to perform the product recovery. For this study, the use of an in situ product recovery method with three combined commercial resins (HP-20, XAD7HP, and HP-2MG) with different chemical properties was performed. A new yeast strain of Saccharomyces cerevisiae was engineered using CRISPR Cas9 (strain EJ2) to deliver heterologous expression of oxygenated acetylated taxanes that are precursors of the anticancer drug Taxol ® (paclitaxel). Microscale cultivations using a definitive screening design (DSD) were set to get the best resin combinations and concentrations to retrieve high taxane titers. Once the best resin treatment was selected by the DSD, semi-continuous cultivation in high throughput microscale was performed to increase the total taxanes yield up to 783 ± 33 mg/L. The best T5α-yl Acetate yield obtained was up to 95 ± 4 mg/L, the highest titer of this compound ever reported by a heterologous expression. It was also observed that by using a combination of the resins in the cultivation, 8 additional uncharacterized taxanes were found in the gas chromatograms compared to the dodecane overlay method. Lastly, the cell-waste reactive oxygen species concentrations from the yeast were 1.5-fold lower in the resin's treatment compared to the control with no adsorbent aid. The possible future implications of this method could be critical for bioprocess intensification, allowing the transition to a semi-continuous flow bioprocess. Further, this new methodology broadens the use of different organisms for natural product synthesis/discovery benefiting from clear bioprocess intensification advantages.

Increased paclitaxel recovery from Taxus baccata vascular stem cells using novel in situ product recovery approaches.

In this study, several approaches were tested to optimise the production and recovery of the widely used anticancer drug Taxol® (paclitaxel) from culturable vascular stem cells (VSCs) of Taxus baccata, which is currently used as a successful cell line for paclitaxel production. An in situ product recovery (ISPR) technique was employed, which involved combining three commercial macro-porous resin beads (HP-20, XAD7HP and HP-2MG) with batch and semi-continuous cultivations of the T. baccata VSCs after adding methyl jasmonate (Me-JA) as an elicitor. The optimal resin combination resulted in 234 ± 23 mg of paclitaxel per kg of fresh-weight cells, indicating a 13-fold improved yield compared to the control (with no resins) in batch cultivation. This resin treatment was further studied to evaluate the resins' removal capacity of reactive oxygen species (ROS), which can cause poor cell growth or reduce product synthesis. It was observed that the ISPR cultivations had fourfold less intracellular ROS concentration than that of the control; thus, a reduced ROS concentration established by the resin contributed to increased paclitaxel yield, contrary to previous studies. These paclitaxel yields are the highest reported to date using VSCs, and this scalable production method could be applied for a diverse range of similar compounds utilising plant cell culture.

Inference on periodicity of circadian time series.

Estimation of the period length of time-course data from cyclical biological processes, such as those driven by the circadian pacemaker, is crucial for inferring the properties of the biological clock found in many living organisms. We propose a methodology for period estimation based on spectrum resampling (SR) techniques. Simulation studies show that SR is superior and more robust to non-sinusoidal and noisy cycles than a currently used routine based on Fourier approximations. In addition, a simple fit to the oscillations using linear least squares is available, together with a non-parametric test for detecting changes in period length which allows for period estimates with different variances, as frequently encountered in practice. The proposed methods are motivated by and applied to various data examples from chronobiology.

Alteraciones neuropsicológicas de un paciente con enfermedad de Parkinson y antecedentes de consumo de sustancias Psicoactivas / Neuropsychological alterations of a patient with Parkinson's disease and background psychoactive substance use

El objetivo de esta investigación fue describir el desempeño cognoscitivo de un paciente de sexo masculino, de 51 años, con 9 años de evolución de Enfermedad de Parkinson (EP) y antecedentes de consumo de sustancias psicoactivas, para lo cual se utilizó un protocolo neuropsicológico encontrando en el test de trazados (TMT-A 24/24 y TMT-B 24/24), prueba de Denominación de Boston (58/60), F.A.S- COWAT (28), Curva de Memoria California (Ascendente), copia (34/36) y evocación (26/36) de la Figura Compleja de Rey Osterrieth, Torre de Londres, Stroop de Colores (C 16", P 31", C-P 57"). Adicionalmente se aplicaron subpruebas del Test de Inteligencia Wechsler para adultos (WAIS-III) (Retención de dígitos 9, sucesión letras-números 10, aritmética 11, dígito-símbolo 10, figuras incompletas 6, diseño con cubos 10, semejanzas 9), el perfil cognoscitivo se encuentra dentro de los parámetros normales para los procesos de lenguaje, memoria, praxias, y habilidades viso-construccionales; se evidencian leves alteraciones en atención, memoria de trabajo y funciones ejecutivas. El consumo prolongado de sustancias psicoactivas es un posible factor de riesgo en el desarrollo temprano de la EP, pero con un perfil cognoscitivo conservado que se puede explicar por su nivel académico previo y su continuo ejercicio intelectual en su profesión, que le exigen constantes actividades cognitivas.

The objective of this research was to describe the cognitive performance of a male patient, aged 51, with 9 years of evolution of Parkinson's disease (PD) and a history of psychoactive substance use, for which a protocol was used neuropsychological finding paths in the test (TMT-A 24/24 and TMT-B 24/24), Boston denomination (58/60), FAS-COWAT (28), Curve Memory California (ascending), copy (34 / 36) and evocation (26/36) of the Rey Osterrieth Complex Figure, Tower of London, Stroop Color (C 16 " W 31", CW 57 "). Additionally applied subtests Wechsler Intelligence Test for Adults (WAIS-III) (retention of digits 9, letter-number sequence 10, arithmetic 11, digit-symbol 10, incomplete figures 6, design with cubes10, similarities 9), cognitive profile is within normal parameters for the processes of language, memory, praxis, visual-constructional and skills, are demonstrated slight alterations in attention, working memory and executive functions. Prolonged use of psychoactive substances is a possible risk factor in early development of PD, but with preserved cognitive profile that can be explained by their prior academic and intellectual exercise continued in his profession, which require constant cognitive activities.