Evaluation of the VITEK 2 Gram-negative (GN) microbial identification test card: collaborative study

A collaborative study was conducted to evaluate the performance of the VITEK 2 Gram-negative (GN) Identification card for use with the VITEK 2 automated microbial identification system. The GN test card is used in the identification of fermenting and nonfermenting Gram-negative bacilli, including the select agent organisms Brucella melitensis, Francisella tularensis, Burkholderia mallei, B. pseudomallei, and Yersinia pestis. The VITEK 2 GN card is based on 47 biochemical tests measuring carbon source utilization, inhibition and resistance, and enzymatic activities. A total of 20 laboratories representing government, industry, and private testing facilities throughout the United States participated. In this study, 720 Gram-negative inclusivity isolates were analyzed by the GN Identification method. Of the 720 well-characterized isolates, 707 were identified correctly, 0 were misidentified, 0 were unidentified, and 13 were not characterized as a Gram-negative organism. Additionally, 120 isolates exclusive of fermenting and nonfermenting Gram-negative bacilli were screened by Gram stain. A total of 117 isolates were correctly excluded. Three organisms were incorrectly characterized by Gram stain procedures, resulting in incorrect analysis and misidentification by VITEK 2 GN. The VITEK 2 GN identification method is an acceptable automated method for the rapid identification of Gram-negative bacteria.     

Analysis of Single‐Molecule Fluorescence Spectroscopic Data with a Markov‐Modulated Poisson Process

We present a photon‐by‐photon analysis framework for the evaluation of data from single‐molecule fluorescence spectroscopy (SMFS) experiments using a Markov‐modulated Poisson process (MMPP). A MMPP combines a discrete (and hidden) Markov process with an additional Poisson process reflecting the observation of individual photons. The algorithmic framework is used to automatically analyze the dynamics of the complex formation and dissociation of Cu²⁺ ions with the bidentate ligand 2,2′‐bipyridine‐4,4′dicarboxylic acid in aqueous media. The process of association and dissociation of Cu²⁺ ions is monitored with SMFS. The dcbpy‐DNA conjugate can exist in two or more distinct states which influence the photon emission rates. The advantage of a photon‐by‐photon analysis is that no information is lost in preprocessing steps. Different model complexities are investigated in order to best describe the recorded data and to determine transition rates on a photon‐by‐photon basis. The main strength of the method is that it allows to detect intermittent phenomena which are masked by binning and that are difficult to find using correlation techniques when they are short‐lived.     

A sequential cyclization/π-extension strategy for modular construction of nanographenes enabled by stannole cycloadditions

The synthesis of polycyclic aromatic hydrocarbons (PAHs) and related nanographenes requires the selective and efficient fusion of multiple aromatic rings. For this purpose, the Diels–Alder cycloaddition has proven especially useful; however, this approach currently faces significant limitations, including the lack of versatile strategies to access annulated dienes, the instability of the most commonly used dienes, and difficulties with aromatization of the [4 + 2] adduct. In this report we address these limitations via the marriage of two powerful cycloaddition strategies. First, a formal Cp₂Zr-mediated [2 + 2 + 1] cycloaddition is used to generate a stannole-annulated PAH. Secondly, the stannoles are employed as diene components in a [4 + 2] cycloaddition/aromatization cascade with an aryne, enabling π-extension to afford a larger PAH. This discovery of stannoles as highly reactive – yet stable for handling – diene equivalents, and the development of a modular strategy for their synthesis, should significantly extend the structural scope of PAHs accessible by a [4 + 2] cycloaddition approach.

Stannoles are introduced as a new, spontaneously aromatizing diene for [4 + 2] cycloadditions that can be easily introduced into diverse conjugated systems, facilitating the efficient synthesis of complex PAHs and their π-extension.     

Structure Elucidation of the First Sex-Inducing Pheromone of a Diatom

Diatoms are abundant unicellular microalgae, responsible for ≈20 % of global photosynthetic CO₂ fixation. Nevertheless, we know little about fundamental aspects of their biology, such as their sexual reproduction. Pheromone-mediated chemical communication is crucial for successful mating. An attraction pheromone was identified in the diatom Seminavis robusta, but metabolites priming cells for sex and synchronizing search and mating behavior remained elusive. These sex-inducing pheromones (SIP) induce cell cycle arrest and trigger the production of the attraction pheromone. Here we describe the challenging structure elucidation of an S. robusta SIP. Guided by metabolomics, a candidate metabolite was identified and elucidated by labeling experiments, NMR, ESI MSⁿ analyses, and chemical transformations. The use of negative ion mode MS was essential to decipher the unprecedented hydroxyproline and β-sulfated aspartate-containing cyclic heptapeptide that acts in femtomolar concentrations.     

Cloud point of poly(N-isopropylacrylamide) synthesized in the presence of cyclodextrin

Polymerization of N-isopropylacrylamide (NIPAAm) was performed in the presence of β-cyclodextrin (β-CD) [or hydroxypropyl-β-cyclodextrin (HOP-β-CD)]. The formation of an inclusion complex between PNIPAAm and β-CD was confirmed by FT-IR, ¹H NMR and 2D-NOESY NMR. It was found that the cloud point (CP) of poly(N-isopropylamide) (PNIPAAm) in the PNIPAAm/β-CD complex in aqueous solution increased up to about 57°C whereas that in the PNIPAAm/β-CD blend is about 33°C. The CP of PNIPAAm in the PNIPAAm/β-CD complex is higher than that in the PNIPAAm/HOP-β-CD inclusion complex. IR and NMR results suggest that PNIPAAm chains are included in CD cavities.