RESUMO
A novel histotripsy approach based on fundamental and second harmonic superposition and incorporating hundred-microsecond-long pulses and two-stage pulse protocol is proposed in this study to rapidly generate mechanically homogenized lesions. Two pulse stages were applied: stage 1, pulses with a pulse duration of 500-600 µs and pulse repetition frequency of 100 Hz, and stage 2, multiple periods, each composed of multiple pulses with the same pulse duration and pulse repetition frequency as those in stage 1, but with an off-time of 600 ms between periods. A custom-designed 1.1/2.2-MHz two-element confocal-annular array, with an f-number of 0.69, and lateral and axial full width at half-maximum pressure dimensions of approximately 1.0 and 6.0 mm, was used. The peak positive/negative pressures at the focus were +22/-7 MPa for 1.1 MHz and +56/-14 MPa with shock wave for 2.2 MHz. To investigate the feasibility of this approach, experiments were designed and performed in tissue-mimicking polyacrylamide gel phantoms with bovine serum albumin and in ex vivo porcine tissues. Cavitation and boiling activities were observed through high-speed photography, and the corresponding acoustic emissions were recorded through passive cavitation detection. Ex vivo experimental results revealed that complete tissue homogeneous regions with regular long tear shape and typical dimensions of 5.80 ± 0.19 mm in axial and 2.20 ± 0.26 mm in lateral were successfully generated in porcine kidney samples. The hematoxylin and eosin staining evidenced that the lesions were thoroughly homogenized and sharply demarcated from untreated regions. These results indicated that the histotripsy approach using fundamental and second harmonic superposition combined with hundred-microsecond pulses and two-stage pulse protocol can efficiently obtain a mechanical disruption of soft tissues with spatial precision, and this approach may have the potential to be developed as a useful tool for precise tumor treatment.
Assuntos
Ablação por Ultrassom Focalizado de Alta Intensidade/métodos , Rim/cirurgia , Litotripsia/métodos , Animais , Desenho de Equipamento , Modelos Animais , Imagens de Fantasmas , SuínosRESUMO
(S)-3-chloro-1-phenyl-1-propanol ((S)-CPPO) is an important chiral intermediate predominantly used in the synthesis of the chiral side chain of (S)-fluoxetine. In this study, carbonyl reductase (CBR) from Novosphingobium aromaticivorans was successfully expressed in recombinant E. coli. The enzymatic activity of the recombinant CBR was significantly increased to 1875â¯U/mL in the fed-batch fermentation in a 10â¯L fermenter and recombinant CBR was then purified and characterized. By regenerating NADH with glucose dehydrogenase, 100â¯g/L 3-chloro-1-phenyl-1-propanone (3-CPP) was successfully converted to (S)-CPPO with a conversion of 100% and ee value of 99.6% after 12â¯h at 30⯰C in PBS buffer (pH 7.0), which are the highest reported to date for the bio-production of (S)-CPPO and presented great potential for green production of (S)-CPPO at industrial scale.