Formaldehyde (FA), in the 0.2-0.4 mM range, is produced and maintained endogenously via enzymatic pathways. At these levels, FA can promote cell proliferation as well as mediate memory formation. Once elevated, FA stress is known to induce cognitive impairments, memory loss, and neurodegeneration owing to its potent DNA and protein cross-linking mechanisms. Optical imaging is a powerful noninvasive approach used to study FA in living systems; however, biocompatible chemical probes for FA are currently lacking. Herein, we report the design, synthesis, and biological evaluation of Formaldehyde Probe 1 (FP1), a new fluorescent indicator based on the 2-aza-Cope sigmatropic rearrangement. The remarkable sensitivity, selectivity, and photostability of FP1 has enabled us to visualize FA in live HEK293TN and Neuroscreen-1 cells. We envision that FP1 will find widespread applications in the study of FA associated with normal and pathological processes.

Cancer stem cells (CSCs) are progenitor cells that contribute to treatment-resistant phenotypes during relapse. CSCs exist in specific tissue microenvironments that cell cultures and more complex models cannot mimic. Therefore, the development of new approaches that can detect CSCs and report on specific properties (e.g., stem cell plasticity) in their native environment have profound implications for studying CSC biology. Herein, we present AlDeSense, a turn-on fluorescent probe for aldehyde dehydrogenase 1A1 (ALDH1A1) and Ctrl-AlDeSense, a matching nonresponsive reagent. Although ALDH1A1 contributes to the detoxification of reactive aldehydes, it is also associated with stemness and is highly elevated in CSCs. AlDeSense exhibits a 20-fold fluorescent enhancement when treated with ALDH1A1. Moreover, we established that AlDeSense is selective against a panel of common ALDH isoforms and exhibits exquisite chemostability against a collection of biologically relevant species. Through the application of surface marker antibody staining, tumorsphere assays, and assessment of tumorigenicity, we demonstrate that cells exhibiting high AlDeSense signal intensity have properties of CSCs. Using these probes in tandem, we have identified CSCs at the cellular level via flow cytometry and confocal imaging, as well as monitored their states in animal models.

The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.

PurposeCataract results from the formation of light-scattering precipitates due to point mutations or accumulated damage in the structural crystallins of the eye lens. Although excised cataracts are predominantly amorphous, in vitro studies show that crystallins are capable of adopting a variety of morphologies depending on the preparation method. Here we characterize thermal, pH-dependent, and UV-irradiated aggregates from wild-type human γS-crystallin (γS-WT) and its aggregation-prone variant, γS-G18V.MethodsAggregates of γS-WT and γS-G18V were prepared under acidic, neutral, and basic pH conditions and held at 25°C or 37°C for 48 hours. UV-induced aggregates were produced by irradiation with a 355-nm laser. Aggregation and fibril formation were monitored via turbidity and thioflavin T (ThT) assays. Aggregates were characterized using intrinsic aromatic fluorescence, powder x-ray diffraction, and mass spectrometry.ResultsγS-crystallin aggregates displayed different characteristics depending on the preparation method. γS-G18V produced a larger amount of detectable aggregates than did γS-WT and at less-extreme conditions. Aggregates formed under basic and acidic conditions yielded elevated ThT fluorescence; however, aggregates formed at low pH did not produce strongly turbid solutions. UV-induced aggregates produced highly turbid solutions but displayed only moderate ThT fluorescence. X-ray diffraction confirms amyloid character in low-pH samples and UV-irradiated samples, although the relative amounts vary.ConclusionsγS-G18V demonstrates increased aggregation propensity compared to γS-WT when treated with heat, acid, or UV light. The resulting aggregates differ in their ThT fluorescence and turbidity, suggesting that at least two different aggregation pathways are accessible to both proteins under the conditions tested.

Photoacoustic imaging is a state-of-the-art technique that combines light excitation with ultrasound generation via the photoacoustic effect. Since sound waves at clinically relevant frequencies undergo minimal perturbation as they pass through the body, photoacoustic imaging is ideal for deep-tissue imaging applications in vivo. Despite its utility in the biomedical field, it is unlikely that undergraduate students will ever experience this technology firsthand in a classroom setting owing to the delicate and expensive nature of the instrumentation. Likewise, students will not be exposed to acoustogenic probes, which are specialized chemicals designed to detect biologically relevant analytes using photoacoustic imaging. With the goal of introducing new chemical analysis and imaging techniques to the undergraduate chemistry curricula, we present a simple and inexpensive setup (all materials were purchased for less than $48 USD) to demonstrate the photoacoustic effect through analyte sensing.