A growing body of literature encourages routine implementation of virtual monoenergetic imaging for dual-energy CT because it affords fewer beam-hardening artifacts, reduced iodine dose, and better... In this article, the authors discuss the technical background and summarize the current body of literature regarding virtual monoenergetic (VM) images derived from dual-energy CT data, which can be...

The methylotrophic yeast Pichia pastoris (Komagataella phaffii) is one of the most commonly used expression systems for heterologous protein production. However the recombination machinery in P. pastoris is less effective in contrast to Saccharomyces cerevisiae, where efficient homologous recombination naturally facilitates genetic modifications. The lack of simple and efficient methods for gene disruption and specifically integrating cassettes has remained a bottleneck for strain engineering in P. pastoris. Therefore tools and methods for targeted genome modifications are of great interest. Here we report the establishment of CRISPR/Cas9 technologies for P. pastoris and demonstrate targeting efficiencies approaching 100%. However there appeared to be a narrow window of optimal conditions required for efficient CRISPR/Cas9 function for this host. We systematically tested combinations of various codon optimized DNA sequences of CAS9, different gRNA sequences, RNA Polymerase III and RNA Polymerase II promoters in combination with ribozymes for the expression of the gRNAs and RNA Polymerase II promoters for the expression of CAS9. Only 6 out of 95 constructs were functional for efficient genome editing. We used this optimized CRISPR/Cas9 system for gene disruption studies, to introduce multiplexed gene deletions and to test the targeted integration of homologous DNA cassettes. This system allows rapid, marker-less genome engineering in P. pastoris enabling unprecedented strain and metabolic engineering applications.

PURPOSE: To evaluate the local tumor control and survival data for magnetic resonance (MR) imaging–guided laser-induced interstitial thermotherapy (LITT) of colorectal liver metastases. MATERIALS AND METHODS: MR imaging–guided LITT was performed in 603 patients (mean age, 61.2 years) with 1,801 liver metastases of colorectal cancer. Survival rates were calculated by means of the Kaplan-Meier method. Local tumor control and tumor volume were evaluated with nonenhanced and contrast material–enhanced MR imaging. Indications for the procedure were defined for patients with five or fewer metastases, none of which were larger than 5 cm in diameter. The indications included recurrent liver metastases after partial liver resection in 37.6% of study patients, metastases in both liver lobes in 32.5%, locally nonresectable lesions in 11.3%, general contraindications for surgery in 4.6%, and refusal to undergo surgical resection in 13.9%. RESULTS: Local recurrence rate at 6-month follow-up was 1.9% (nine of 474) for ...

The efficacy of microwave ablation is mainly determined by preablation tumor size and location in relation to the hilum. PurposeTo prospectively evaluate the safety and efficacy of microwave ablation therapy of unresectable pulmonary metastases.Materials and MethodsAll patients provided informed consent for this prospective institutional review board–approved study. Eighty patients (30 men, 50 women; mean age, 59.7 years ± 6.4; range, 48–68 years) underwent computed tomography–guided percutaneous microwave ablation of pulmonary metastatic lesions in 130 sessions. The tumors represented metastases from colorectal carcinoma, breast carcinoma, hepatocellular carcinoma, renal cell carcinoma, and bronchogenic carcinoma; there was no evidence of extrapulmonary metastasis for any tumor. Logistic regression analysis was used for evaluation of the statistical significance of factors affecting the end result of microwave ablation therapy. The Kaplan-Meier method was used for estimation of survival rates.ResultsComplete, successful ablation was achieved in 95 (73.1%) of 130 lesions. Successful tumor ablation was significantly more fre...