New insight into the role of the β3 subunit of the GABA<Subscript>A</Subscript>-R in development,behavior, body weight regulation,and anesthesia revealed by conditional gene knockout

Background  

The β3 subunit of the γ-aminobutyric acid type A receptor (GABA_A-R) has been reported to be important for palate formation, anesthetic action, and normal nervous system function. This subunit has also been implicated in the pathogenesis of Angelman syndrome and autism spectrum disorder. To further investigate involvement of this subunit, we previously produced mice with a global knockout of β3. However, developmental abnormalities, compensation, reduced viability, and numerous behavioral abnormalities limited the usefulness of that murine model. To overcome many of these limitations, a mouse line with a conditionally inactivated β3 gene was engineered.     

Prevalence and Significance of Approximate Symmetry in Organic Pc Structures

A survey has been made of the well determined (R≤0.050) organic crystal structures that have only a single glide plane (e. g., are in space group Pc) and that have more than one crystallographically independent formula unit (Z′>1); the goal was to discover what fraction have additional approximate symmetry. Of the 377 unique structures examined 8 % should almost certainly been refined in a higher-symmetry unit cell; 86 % of the remaining 347 have approximate symmetry that is periodic in at least one dimension. While these percentages are similar to those found for P1 structures (C. P. Brock, Acta Crystallogr., Sect. B 2022 , 78, 576–588), the types of approximate symmetry differ because 89 % of the P1, Z′>1 structures were composed of enantiopure material. The 347 reliable Pc structures include 118 that are slightly distorted or mimicked Cc and P2₁/c structures, 15 of which were reported as having been determined at room temperature. The distortions in another 72 are so large that the approximate symmetry must be seen as periodic in two dimensions only. These results suggest that symmetry lowering may accompany transformation of a crystal nucleus to a macroscopic crystal.     

Sorted cell microarrays as platforms for high-content informational bioassays

We report on surface-engineered microarrays that provide in situ cell sorting, localization, and immobilization of various subsets of human primary lymphocytes, followed by an on-chip bioassay for ionizing-radiation-induced cytogenetic damage. The microarray format eliminates the necessity of separating cell sub-populations by alternative means (such as fluorescence- or magnetic-activated cell sorting) prior to performing informational bioassays. To exemplify the potential of this on-chip cytometry approach, we have integrated the cytokinesis-block micronucleus cytome (CBMNcyt) assay with the microarray platform for analysis of the chromosome damage profile of specific subsets of human peripheral lymphocytes. Microarray results were compared with data obtained from the traditional CBMNcyt assay on heterogeneous lymphocyte populations, and with flow cytometry data. Our results suggest that cytogenetic damage caused by ionizing radiation is not uniformly distributed across all lymphocytes subsets, but rather concentrated in specific subsets. The salient features of our approach are that it requires very small volumes of reagents, allows sorting of lymphocyte subsets in situ, increases parallelism of cell assays and is amenable to high content microscopy analysis. The on-chip cytometry format opens new vistas for advanced cell-based assays, potentially bringing to light important information which remains hidden with conventional assays and hence engendering new discoveries in cell biology.     

A comparison of positive and negative ion collision‐induced dissociation for model heptapeptides with one basic residue

The effects of the identity and position of basic residues on peptide dissociation were explored in the positive and negative modes. Low‐energy collision‐induced dissociation (CID) was performed on singly protonated and deprotonated heptapeptides of the type: XAAAAAA, AAAXAAA, AAAAAXA and AAAAAAX, where X is arginine (R), lysine (K) or histidine (H) residues and A is alanine. For [M + H]⁺, the CID spectra are dominated by cleavages adjacent to the basic residues and the majority of the product ions contain the basic residues. The order of a basic residue's influence on fragmentation of [M + H]⁺ is arginine > histidine ≈ lysine, which is also the order of decreasing gas‐phase basicity for these amino acids. These results are consistent with the side chains of basic residues being positive ion charge sites and with the more basic arginine residues having a higher retention (i.e. sequestering) of the positive charge. In contrast, for [M ? H]^? the identity and position of basic residues has almost no effect on backbone fragmentation. This is consistent with basic residues not being negative mode charge sites. For these peptides, more complete series of backbone fragments, which are important in the sequencing of unknowns, can be found in the negative mode. Spectra at both polarities contain C‐terminal y‐ions, but y_n″⁺ has two more hydrogens than the corresponding y_n^?. Another major difference is the production of the N‐terminal backbone series b_n⁺ in the positive mode and c_n^? in the negative mode. Thus, comparison of positive and negative ion spectra with an emphasis on searching for pairs of ions that differ by 2 Da (y_n″⁺ vs y_n^?) and by 15 Da (b_n⁺ vs c_n^?) may be a useful method for determining whether a product ion is generated from the C‐terminal or the N‐terminal end of a peptide. In addition, a characteristic elimination of NH?C?NH from arginine residues is observed for deprotonated peptides. Copyright © 2010 John Wiley & Sons, Ltd.     

Auditory evoked potentials in two short-finned pilot whales (Globicephala macrorhynchus)

The hearing sensitivities of two short-finned pilot whales (Globicephala macrorhynchus) were investigated by measuring auditory evoked potentials generated in response to clicks and sinusoidal amplitude modulated (SAM) tones. The first whale tested, an adult female, was a long-time resident at SeaWorld San Diego with a known health history. Click-evoked responses in this animal were similar to those measured in other echolocating odontocetes. Auditory thresholds were comparable to dolphins of similar age determined with similar evoked potential methods. The region of best sensitivity was near 40 kHz and the upper limit of functional hearing was between 80 and 100 kHz. The second whale tested, a juvenile male, was recently stranded and deemed non-releasable. Click-evoked potentials were not detected in this animal and testing with SAM tones suggested severe hearing loss above 10 kHz.     

Sign patterns that require real, nonreal or pure imaginary eigenvalues

We characterize the n-by-n sign pattern matrices that require all real, all nonreal, and all pure imaginary eigenvalues. Characterization of sign patterns that allow a real eigenvalue and those that allow a nonreal eigenvalue then follow. Some related specialized results and a characterization of sign patterns that allow a positive real eigenvalue are included.     

MR Imaging of metastatic pancreatic VIPoma

The MR findings in a 32-year-old man with pancreatic VIPoma and liver metastases are described. A 2-cm mass was present in the region of the tail of the pancreas that was best shown on T₁-weighted fat-suppressed images as a low-signal intensity mass. Multiple liver metastases were present that showed intense peripheral ring enhancement on immediate post gadolinium spoiled gradient echo images.