Mikrosfere ropinirol hidroklorida za polagano oslobađanje: Utjecaj procesnih parametara

An emulsion solvent evaporation method was employed to prepare microspheres of ropinirole hydrochloride, a highly water soluble drug, by using ethylcellulose and PEG with the help of 32 full factorial design. The microspheres were made by incorporating the drug in a polar organic solvent, which was emulsified using liquid paraffin as an external oil phase. Effects of various process parameters such as viscosity of the external phase, selection of the internal phase, surfactant selection and selection of stirring speed were studied. Microspheres were evaluated for product yield, encapsulation efficiency and particle size. Various drug/ethylcellulose ratios and PEG concentrations were assayed. In vitro dissolution profiles showed that ethylcellulose microspheres were able to control release of the drug for a period of 12 h.Mikrosfere ropinirol hidroklorida, ljekovite tvari vrlo dobro topljive u vodi, pripravljene su metodom isparavanja otapala, koristeći etilcelulozu i PEG te 32 potpuno faktorijalno dizajniranje. Mikrosfere su pripravljene na sljedeći način: otopina ljekovite tvari u polarnom organskom otapalu emulgirana je s tekućim parafinom kao vanjskom uljnom fazom. Ispitivan je utjecaj različitih procesnih parametara poput viskoznosti vanjske faze, vrste interne faze i površinski aktivne tvari te brzine miješanja. Za pripravljene mikrosfere određeno je iskorištenje, učinkovitost inkapsuliranja i veličina čestica. Isprobavani su različiti odnosi ljekovite tvari i etilceluloze te koncentracija PEG-a. In vitro pokusi su pokazali da je oslobađanje ljekovite tvari kontrolirano tijekom 12 h

Familial retinoblastoma due to intronic LINE-1 insertion causes aberrant and noncanonical mRNA splicing of the RB1 gene

Retinoblastoma (RB, MIM 180200) is the paradigm of hereditary cancer. Individuals harboring a constitutional mutation in one allele of the RB1 gene have a high predisposition to develop RB. Here, we present the first case of familial RB caused by a de novo insertion of a full-length long interspersed element-1 (LINE-1) into intron 14 of the RB1 gene that caused a highly heterogeneous splicing pattern of RB1 mRNA. LINE-1 insertion was inferred by mRNA studies and full-length sequenced by massive parallel sequencing. Some of the aberrant mRNAs were produced by noncanonical acceptor splice sites, a new finding that up to date has not been described to occur upon LINE-1 retrotransposition. Our results clearly show that RNA-based strategies have the potential to detect disease-causing transposon insertions. It also confirms that the incorporation of new genetic approaches, such as massive parallel sequencing, contributes to characterize at the sequence level these unique and exceptional genetic alterations.This study was funded by grants of the Instituto de Salud Carlos III (PI12/00816 and RTICC RD12/0036/0027). CR-M was supported by a MINNECO contract. FC was supported by Asociación Pablo Ugarte and Miguelañez SA. SM was supported by a CIBERER contract. We greatly appreciate the collaboration of the RB patients, their parents and their families.S

Succinic acid production from wheat using a biorefining strategy

The biosynthesis of succinic acid from wheat flour was investigated in a two-stage bioprocess. In the first stage, wheat flour was converted into a generic microbial feedstock either by fungal fermentation alone or by combining fungal fermentation for enzyme and fungal bio-mass production with subsequent flour hydrolysis and fungal autolysis. In the second stage, the generic feedstock was converted into succinic acid by bacterial fermentation by Actinobacillus succinogenes. Direct fermentation of the generic feedstock produced by fungal fermentation alone resulted in a lower succinic acid production, probably due to the low glucose and nitrogen concentrations in the fungal broth filtrate. In the second feedstock production strategy, flour hydrolysis conducted by mixing fungal broth filtrate with wheat flour generated a glucose-rich stream, while the fungal bio-mass was subjected to autolysis for the production of a nutrient-rich stream. The possibility of replacing a commercial semi-defined medium by these two streams was investigated sequentially. A. succinogenes fermentation using only the wheat-derived feedstock resulted in a succinic acid concentration of almost 16 g l–1 with an overall yield of 0.19 g succinic acid per g wheat flour. These results show that a wheat-based bio-refinery employing coupled fungal fermentation and subsequent flour hydrolysis and fungal autolysis can lead to a bacterial feedstock for the efficient production of succinic acid.The biosynthesis of succinic acid from wheat flour was investigated in a two-stage bio-process. In the first stage, wheat flour was converted into a generic microbial feedstock either by fungal fermentation alone or by combining fungal fermentation for enzyme and fungal bio-mass production with subsequent flour hydrolysis and fungal autolysis. In the second stage, the generic feedstock was converted into succinic acid by bacterial fermentation by Actinobacillus succinogenes. Direct fermentation of the generic feedstock produced by fungal fermentation alone resulted in a lower succinic acid production, probably due to the low glucose and nitrogen concentrations in the fungal broth filtrate. In the second feedstock production strategy, flour hydrolysis conducted by mixing fungal broth filtrate with wheat flour generated a glucose-rich stream, while the fungal bio-mass was subjected to autolysis for the production of a nutrient-rich stream. The possibility of replacing a commercial semi-defined medium by these two streams was investigated sequentially. A. succinogenes fermentation using only the wheat-derived feedstock resulted in a succinic acid concentration of almost 16 g l–1 with an overall yield of 0.19 g succinic acid per g wheat flour. These results show that a wheat-based biorefinery employing coupled fungal fermentation and subsequent flour hydrolysis and fungal autolysis can lead to a bacterial feedstock for the efficient production of succinic acid