Studies of the biosynthesis of Sarubicin A

Early studies on the biosynthesis of Sarubicin A, 1, had suggested the intermediacy of the novel aromatic amino acid 6-hydroxyanthranilic acid, 17. To investigate the biosynthetic steps beyond 17, [¹³C-¹⁵N carboxamide] 6-hydroxyanthranilamide, 27a, was synthesized, and fed to fermentations of Streptomyces helicus. Incorporation of 27a into 1 provided suggestive evidence that 27 is a new product of the shikimate pathway, and provided evidence that amide formation occurs early in the biosynthetic pathway to 1. Several possibilities exist for the timing of the hydroxylation and Cglycosylation steps of the quinone ring of 1. To test the chemical viability of the potential intermediate 5- glycosyl-3,6-dihydroxyanthranilamide, 29, the model quinione 67 was synthesized and reduced. The reduction product, 68, was stable in the absence of 0₂, suggesting that the proposed C-glycoside 29 is a plausible intermediate. A model for 29 was synthesized via an ultrasound catalyzed Diels- Alder reaction between the 2,2-dimethy1-5,8-dimethoxy-1,2,3,4-tetrahydro- 4-oxoquinazoline-6-carboxaldehyde, 71, and the diene 72, to give the pyran derivative 73. However, attempts to construct the carbon skeleton of 2 9 with diene 74 were unsuccessful. Isochorismate synthase was isolated from Enterobacter aerogenes 62-1, partially purified, and incubated with chorismic acid, 19, in H₂¹⁸0 . The product, isochorismic acid, 18, was purified and found to contain ¹⁸0 at the C-2 hydroxyl, demonstrating that the enzyme-catalyzed reaction proceeds via an SN₂' or addition-elimination type of reaction

A Cluster of Trace-Concentration Methamphetamine Identifications in Racehorses Associated with a Methamphetamine-Contaminated Horse Trailer: A Report and Analysis

Three low concentration methamphetamine “positive” tests were linked to use of a methamphetamine-contaminated trailer to transport the affected horses. This incident establishes methamphetamine as a human-use substance that can inadvertently enter the environment of racing horses, resulting in urinary methamphetamine “positives;” an interim regulatory cut-off of 15 ng/mL for methamphetamine in post-race urine is proposed

Trichomonas vaginalis Detection in Urogenital Specimens from Symptomatic and Asymptomatic Men and Women by Use of the cobas TV/MG Test

Trichomonas vaginalis is a prevalent sexually transmitted infection (STI). Diagnosis has historically relied on either microscopic analysis or culture, the latter being the previous gold standard. However, these tests are not readily available for male diagnosis, generally only perform well for symptomatic women, and are not as sensitive as nucleic acid amplification tests (NAATs). Men are largely asymptomatic but carry the organism and transmit to their sexual partners. This multicenter, prospective study evaluated the performance of the cobas T. vaginalis/Mycoplasma genitalium (TV/MG) assay for detection of T. vaginalis DNA compared with patient infection status (PIS) defined by a combination of commercially available NAATs and culture using urogenital specimens. A total of 2,064 subjects (984 men and 1,080 women, 940 [45.5%] symptomatic, 1,124 [54.5%] asymptomatic) were evaluable. In women, sensitivity ranged from 99.4% (95% confidence interval [CI] 96.8 to 99.9%) using vaginal samples to 94.7% (95% CI 90.2 to 97.2%) in PreservCyt samples. Specificity ranged from 98.9 to 96.8% (95% CI 95.4 to 97.8%). In men, the cobas TV/MG assay was 100% sensitive for the detection of T. vaginalis in both male urine samples and meatal swabs, with specificity of 98.4% in urine samples and 92.5% in meatal swabs. The cobas TV/MG is a suitable diagnostic test for the detection of T. vaginalis, which could support public health efforts toward infection control and complement existing STI programs

Empirical Legal Studies Before 1940: A Bibliographic Essay

The modern empirical legal studies movement has well-known antecedents in the law and society and law and economics traditions of the latter half of the 20th century. Less well known is the body of empirical research on legal phenomena from the period prior to World War II. This paper is an extensive bibliographic essay that surveys the English language empirical legal research from approximately 1940 and earlier. The essay is arranged around the themes in the research: criminal justice, civil justice (general studies of civil litigation, auto accident litigation and compensation, divorce, small claims, jurisdiction and procedure, civil juries), debt and bankruptcy, banking, appellate courts, legal needs, legal profession (including legal education), and judicial staffing and selection. Accompanying the essay is an extensive bibliography of research articles, books, and reports

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Evaluation of the Performance of the Cobas CT/NG Test for Use on the Cobas 6800/8800 Systems for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Male and Female Urogenital Samples

The clinical performance of the Cobas CT/NG assay on the Cobas 6800/8800 systems (Cobas) for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae was established in a multisite, prospective collection study using male and female urogenital specimens; supportive data from archived specimens were also included. The results obtained with the Cobas assay were compared with the patient infected status derived from a combination of U.S. Food and Drug Administration-approved nucleic acid amplification tests to determine the sensitivity and specificity of detection from each sample type. The sensitivity of Cobas for the detection of C. trachomatis in female specimens was 95.6% (95% confidence interval [CI], 92.4% to 97.4%) for urine; 98.6% (95% CI, 95.2% to 99.6%) and 99.2% (95% CI, 95.4% to 99.9%) for clinician- and self-collected vaginal swab specimens, respectively; 93.3% (95% CI, 89.6% to 95.7%) for endocervical swabs; and 92.5% (95% CI, 88.7% to 95.1%) for cervical swab samples in PreservCyt. The specificity for the detection of C. trachomatis was ≥98.8% for all female sample types. Sensitivity and specificity estimates of Cobas for the detection of C. trachomatis in male urine samples were 100% (96.8% to 100.0%) and 99.7% (95% CI, 99.2% to 99.9%), respectively. The sensitivity of Cobas for the detection of N. gonorrhoeae in female specimens was 94.8% (95% CI, 89.6% to 97.4%) for urine; 100.0% (95% CI, 87.9% to 100.0%) and 100.0% (95% CI, 87.9% to 100.0%) for clinician- and self-collected vaginal swab specimens, respectively; 97.0% (95% CI, 91.5% to 99.0%) for endocervical swabs; and 96.6% (95% CI, 90.6% to 98.8%) for cervical samples in PreservCyt; the specificity for all female sample types was >99.0%. The sensitivity and specificity of Cobas for detecting N. gonorrhoeae in male urine were 100.0% (95% CI, 95.8% to 100.0%) and 99.5% (95% CI, 98.8% to 99.8%), respectively. Fully automated assays help fill the clinical need for a sensitive, high-throughput screening tool to aid public health efforts to control C. trachomatis and N. gonorrhoeae infections

Logging and indigenous hunting impacts on persistence of large Neotropical animals

Areas allocated for industrial logging and community-owned forests account for over 50% of all remaining tropical forests. Landscapescale conservation strategies that include these forests are expected to have substantial benefits for biodiversity, especially for large mammals and birds that require extensive habitat but that are susceptible to extirpation due to synergies between logging and hunting. In addition, their responses to logging alone are poorly understood due to their cryptic behavior and low densities. In this study, we assessed the effects of logging and hunting on detection and occupancy rates of large vertebrates in a multiple- use forest on the Guiana Shield. Our study site was certified as being responsibly managed for timber production and indigenous communities are legally guaranteed use-rights to the forest. We coupled camera-trap data for wildlife detection with a spatially explicit dataset on indigenous hunting. A multi-species occupancy model found a weak positive effect of logging on occupancy and detection rates, while hunting had a weak negative effect. Model predictions of species richness were also higher in logged forest sites compared to unlogged forest sites. Density estimates for jaguars and ocelots in our multiple- use area were similar to estimates reported for fully protected areas. Involvement of local communities in forest management, control of forest access, and nesting production forests in a landscape that includes protected areas seemed important for these positive biodiversity outcomes. The maintenance of vertebrate species bodes well for both biodiversity and the humans that depend on multiple- use forests