Purification and Characterization of Acinetobacter calcoaceticus 4-Hydroxybenzoate 3-Hydroxylase after Its Overexpression in Escherichia coli

4-Hydroxybenzoate 3-hydroxylase [EC 1.14.13.2] from Acinetobacter calcoaceticus was purified to homogeneity following the 40-fold overexpression of this gene (pobA) in Escherichia coli. Overexpression was accomplished by placing the folA gene (encoding trimethoprim-resistant dihydrofolate reductase) directly downstream of the pobA gene, and demanding growth of recombinants on elevated concentration of trimethoprim. Presumably, the surviving variants have undergone a genetic alteration which allowed the overexpression of both folA and pobA. 4-Hydroxybenzoate 3-hydroxylase was purified in two chromatographic steps, characterized biochemically, and its properties were compared to those of its homolog from Pseudomonas fluorescens. The two enzymes differ in their reponse to Cl− ion inhibition. A single ami no acid change in the putative NADPH-binding site is proposed to account for this difference. The inhibitory and catalytic properties of substrate analogs were also examine

The Virtues of Abstention: Separation of Powers in Al-Nashiri II

(Excerpt) Part I examines various scholarly approaches to judicial deference, then considers deference in the context of military commissions. In Part II, the history of military commissions in the United States is examined, paying particular attention to the extended dialogue among the coordinate federal branches that created the system currently in operation. The decision in Al-Nashiri II not to adjudicate a collateral attack on one of these commissions is the focus of Part III. That Part embraces the underlying jurisdictional challenge at stake in Al-Nashiri II, the development of abstention doctrine generally and as applied to the current commissions, as well as the role judicial deference played in the panel’s decision. Finally, in Part IV, this Note argues that the path of abstention had many virtues in this case and as a rule of law, because it furthered sound separation of powers principles by respecting the considered judgments of Congress and successive Presidents. Part IV first categorizes the type of deference the panel engaged in by abstaining. Next, it considers the effect the decision will have on future collateral attacks on commission proceedings, as federal courts will now review military commission final judgments, just as Congress and the President intended, rather than intervening indiscriminately. This Note argues that this effect will, in turn, preserve the commission system created by Congress—the branch best suited to weigh the intricate national security considerations involved in prosecuting and bringing to justice those who, in their attempt to thwart our military effort, violate the laws of war

Christian Legal Thought

(Excerpt) On January 26, 2018, the Journal of Catholic Legal Studies proudly hosted a conference on Christian Legal Thought: Materials and Cases a first-of-its-kind casebook authored by Patrick M. Brennan and William S. Brewbaker. Held in Manhattan at the New York Athletic Club, the conference brought together scholars from law schools across the country to discuss the casebook’s impact, as well as the role Christian legal thought might play in the contemporary law school curriculum

Duration of androgen deprivation therapy with postoperative radiotherapy for prostate cancer: a comparison of long-course versus short-course androgen deprivation therapy in the RADICALS-HD randomised trial

Background Previous evidence supports androgen deprivation therapy (ADT) with primary radiotherapy as initial treatment for intermediate-risk and high-risk localised prostate cancer. However, the use and optimal duration of ADT with postoperative radiotherapy after radical prostatectomy remains uncertain. Methods RADICALS-HD was a randomised controlled trial of ADT duration within the RADICALS protocol. Here, we report on the comparison of short-course versus long-course ADT. Key eligibility criteria were indication for radiotherapy after previous radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to add 6 months of ADT (short-course ADT) or 24 months of ADT (long-course ADT) to radiotherapy, using subcutaneous gonadotrophin-releasing hormone analogue (monthly in the short-course ADT group and 3-monthly in the long-course ADT group), daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as metastasis arising from prostate cancer or death from any cause. The comparison had more than 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 75% to 81% (hazard ratio [HR] 0·72). Standard time-to-event analyses were used. Analyses followed intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov , NCT00541047 . Findings Between Jan 30, 2008, and July 7, 2015, 1523 patients (median age 65 years, IQR 60–69) were randomly assigned to receive short-course ADT (n=761) or long-course ADT (n=762) in addition to postoperative radiotherapy at 138 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 8·9 years (7·0–10·0), 313 metastasis-free survival events were reported overall (174 in the short-course ADT group and 139 in the long-course ADT group; HR 0·773 [95% CI 0·612–0·975]; p=0·029). 10-year metastasis-free survival was 71·9% (95% CI 67·6–75·7) in the short-course ADT group and 78·1% (74·2–81·5) in the long-course ADT group. Toxicity of grade 3 or higher was reported for 105 (14%) of 753 participants in the short-course ADT group and 142 (19%) of 757 participants in the long-course ADT group (p=0·025), with no treatment-related deaths. Interpretation Compared with adding 6 months of ADT, adding 24 months of ADT improved metastasis-free survival in people receiving postoperative radiotherapy. For individuals who can accept the additional duration of adverse effects, long-course ADT should be offered with postoperative radiotherapy. Funding Cancer Research UK, UK Research and Innovation (formerly Medical Research Council), and Canadian Cancer Society

The Virtues of Abstention: Separation of Powers in Al-Nashiri II

(Excerpt) Part I examines various scholarly approaches to judicial deference, then considers deference in the context of military commissions. In Part II, the history of military commissions in the United States is examined, paying particular attention to the extended dialogue among the coordinate federal branches that created the system currently in operation. The decision in Al-Nashiri II not to adjudicate a collateral attack on one of these commissions is the focus of Part III. That Part embraces the underlying jurisdictional challenge at stake in Al-Nashiri II, the development of abstention doctrine generally and as applied to the current commissions, as well as the role judicial deference played in the panel’s decision. Finally, in Part IV, this Note argues that the path of abstention had many virtues in this case and as a rule of law, because it furthered sound separation of powers principles by respecting the considered judgments of Congress and successive Presidents. Part IV first categorizes the type of deference the panel engaged in by abstaining. Next, it considers the effect the decision will have on future collateral attacks on commission proceedings, as federal courts will now review military commission final judgments, just as Congress and the President intended, rather than intervening indiscriminately. This Note argues that this effect will, in turn, preserve the commission system created by Congress—the branch best suited to weigh the intricate national security considerations involved in prosecuting and bringing to justice those who, in their attempt to thwart our military effort, violate the laws of war

Christian Legal Thought

(Excerpt) On January 26, 2018, the Journal of Catholic Legal Studies proudly hosted a conference on Christian Legal Thought: Materials and Cases a first-of-its-kind casebook authored by Patrick M. Brennan and William S. Brewbaker. Held in Manhattan at the New York Athletic Club, the conference brought together scholars from law schools across the country to discuss the casebook’s impact, as well as the role Christian legal thought might play in the contemporary law school curriculum