The Nemourinae (Insecta, Nemouridae) of the eastern Nearctic.

Morphology-based concepts of the 26 eastern Nearctic species of the subfamily Nemourinae (Insecta, Nemouridae) Billberg, 1820 are reviewed. Nemourinae is represented in this region by Nemoura Latrielle, 1796, Ostrocerca Ricker, 1952, Paranemoura Needham & Claassen, 1925, Podmosta Ricker, 1952, Prostoia Ricker, 1952, Shipsa Ricker, 1952, Soyedina Ricker, 1952, and Zapada Ricker, 1952. Wing venation and patterns of mottling are depicted with standard light microscopy. Diagnostic external reproductive structures are emphasized with line drawings, scanning electron microscopy, and standard light microscopy. A genus-level key to male and female adults is included. Species-level keys for Ostrocerca, Paranemoura, Prostoia, Soyedina, and Zapada are also provided.

Dracoleuctra, a new wingless stonefly genus from southern Oregon, U.S.A. (Plecoptera: Leuctridae).

Dracoleuctra siskiyou Lee Baumann gen. nov. sp. nov. a new apterous genus and species in the stonefly family Leuctridae (Insecta, Plecoptera), is described from males and females collected in southern Oregon, U.S.A. Habitus and terminalia are illustrated using both light photography images and scanning electron micrographs. Diagnostic characters, habitat, biological notes, and stonefly apterism in the Klamath Mountains region are discussed.

Alloperla clarki sp. nov. (Plecoptera: Chloroperlidae), a new species from the eastern Nearctic with discussion of a new species group.

Alloperla Banks, 1906 (Plecoptera: Chloroperlidae) is a speciose genus of spring- and summer-emergent stoneflies known from the Oriental, eastern Palearctic, and Nearctic realms. Over 50 species are currently recognized. Alloperla clarki sp. nov. is described herein from the adult male stage from a small geographic area in the Appalachian Mountains in Virginia and West Virginia, USA. Diagnostic characters are presented with scanning electron microscope (SEM) photomicrographs. Alloperla clarki sp. nov. most closely resembles A. biserrata Nelson Kondratieff, 1980, A. nanina Banks, 1911, and A. stipitata Surdick, 2004. These four species share similar characteristics of the male epiproct and appear to comprise a species group. The Alloperla nanina Group is formally proposed. Comparative SEM images are also provided for A. biserrata, A. nanina, and A. stipitata to assist with differentiating between these four species.

Soyedina Ricker, 1952 (Plecoptera: Nemouridae) in the eastern Nearctic: review of species concepts, proposed morphology-based species groups, and description of a new species from North Carolina.

The eastern Nearctic species of the genus Soyedina Ricker, 1952 (Plecoptera: Nemouridae) are reviewed. Two morphology-based species groups are proposed based on epiproct characteristics. Soyedina sheldoni sp. nov. is described from the southern Appalachian Highland region of western North Carolina. A distribution map and a dichotomous key to all nine Nearctic species are provided.

Atlas of Ohio Aquatic Insects: Volume II, Plecoptera.

BACKGROUND: We provide volume II of a distributional atlas of aquatic insects for the eastern USA state of Ohio. This treatment of stoneflies (Plecoptera) is companion to Armitage et al. (2011) on caddisflies (Trichoptera). We build on a recent analysis of Ohio stonefly diversity patterns based on large drainages (DeWalt et al. 2012), but add 3717 new records to the data set. We base most analyses on the United States Geological Survey Hierarchical Unit Code eight (HUC8) drainage scale. In addition to distributional maps for each species, we provide analyses of species richness versus HUC8 drainage area and the number of unique locations in a HUC8 drainage, species richness versus Ohio counties, analyze adult presence phenology throughout the year, and demonstrate stream size range affiliation for each species. NEW INFORMATION: This work is based on a total of 7797 specimen records gathered from 21 regional museums, agency data, personal collections, and from the literature Table 1. To our knowledge this is the largest stonefly data set available for a similarly sized geopolitical area anywhere in the world. These data are made available as a Darwin Core Archive supported by the Pensoft Integrated Publishing Toolkit (DeWalt et al. 2016b). All known published papers reporting stoneflies from Ohio are detailed in Suppl. material 1. We recovered 102 species from Ohio, including all nine Nearctic families Table 2​. Two species were removed from the DeWalt et al. (2012) list and two new state records added. Perlidae (32 spp.) was most speciose, compared to the low diversity Pteronarcyidae (2 spp.) and Peltoperlidae (1 sp.). The richest HUC8 drainages occurred in northeastern, south-central, and southern regions of the state where drainages were heavily forested, had the highest slopes, and were contained within or adjacent to the unglaciated Allegheny and Appalachian Plateaus. Species poor drainages occurred mainly in the northwestern region where Wisconsinan aged lake plains climaxed to an expansive wooded wetland, the Black Swamp. The unglaciated Lower Scioto drainage (72 spp.) in south-central Ohio supported the greatest species richness. There was no relationship between species richness and HUC8 drainage size, but the number of unique locations in a drainage strongly related to species richness. All Ohio counties were represented in the data set with Hocking County (59 spp.) of the Lower Scioto drainage being the richest and most heavily sampled. Adult presence phenology was influenced by phylogenetic relationships such that the superfamily Nemouroidea (Capniidae, Leuctridae, Nemouridae, and Taeniopterygidae) generally emerged in winter and spring while the superfamilies Pteronarcyoidea (Pteronarcyidae, Peltoperlidae) and Perloidea (Chloroperlidae, Perlidae, Perlodidae) emerged later, some species continuing emergence through summer months. Species often occupied specific stream size ranges, while others were generalists. Two species once histrorically abundant in the western Lake Erie Bass Islands no longer reside there. Each of the 102 species is discussed in detail, including several that require additional collecting efforts to confirm their identities, presence, and distribution in Ohio.

A review of the Nearctic genus Prostoia (Ricker) (Plecoptera, Nemouridae), with the description of a new species and a surprising range extension for P. hallasi Kondratieff & Kirchner.

The Nearctic genus Prostoia (Plecoptera: Nemouridae) is reviewed. Prostoia ozarkensis sp. n. is described from the male and female adult stages mainly from the Interior Highland region encompassing portions of Arkansas, Missouri, and Oklahoma. Prostoia ozarkensis sp. n. appears most closely related to two species, one distributed broadly across the western Nearctic region, P. besametsa (Ricker), and one found widely throughout the central and eastern Nearctic regions, P. completa (Walker). A surprising range extension is noted for P. hallasi Kondratieff & Kirchner, a species once known only from the Great Dismal Swamp, from small upland streams in southern Illinois. Additional new state records are documented for P. besametsa, P. completa, P. hallasi and P. similis (Hagen). Taxonomic keys to Prostoia males and females are provided, and scanning electron micrographs of adult genitalia of all species are given.

Time-dependent changes in serum biomarker levels after blast traumatic brain injury.

Neuronal and glial proteins detected in the peripheral circulating blood after injury can reflect the extent of the damage caused by blast traumatic brain injury (bTBI). The temporal pattern of their serum levels can further predict the severity and outcome of the injury. As part of characterizing a large-animal model of bTBI, we determined the changes in the serum levels of S100B, neuron-specific enolase (NSE), myelin basic protein (MBP), and neurofilament heavy chain (NF-H). Blood samples were obtained prior to injury and at 6, 24, 72 h, and 2 weeks post-injury from animals with different severities of bTBI; protein levels were determined using reverse phase protein microarray (RPPM) technology. Serum levels of S100B, MBP, and NF-H, but not NSE, showed a time-dependent increase following injury. The detected changes in S100B and MBP levels showed no correlation with the severity of the injury. However, serum NF-H levels increased in a unique, rapid manner, peaking at 6 h post-injury only in animals exposed to severe blast with poor clinical and pathological outcomes. We conclude that the sudden increase in serum NF-H levels following bTBI may be a useful indicator of injury severity. If additional studies verify our findings, the observed early peak of serum NF-H levels can be developed into a useful diagnostic tool for predicting the extent of damage following bTBI.

Selection of cholera toxin specific IgNAR single-domain antibodies from a naïve shark library.

Shark immunoglobulin new antigen receptor (IgNAR, also referred to as NAR) variable domains (Vs) are single-domain antibody (sdAb) fragments containing only two hypervariable loop structures forming 3D topologies for a wide range of antigen recognition and binding. Their small size ( approximately 12kDa) and high solubility, thermostability and binding specificity make IgNARs an exceptional alternative source of engineered antibodies for sensor applications. Here, two new shark NAR V display libraries containing >10(7) unique clones from non-immunized (naïve) adult spiny dogfish (Squalus acanthias) and smooth dogfish (Mustelus canis) sharks were constructed. The most conserved consensus sequences derived from random clone sequence were compared with published nurse shark (Ginglymostoma cirratum) sequences. Cholera toxin (CT) was chosen for panning one of the naïve display libraries due to its severe pathogenicity and commercial availability. Three very similar CT binders were selected and purified soluble monomeric anti-CT sdAbs were characterized using Luminex(100) and traditional ELISA assays. These novel anti-CT sdAbs selected from our newly constructed shark NAR V sdAb library specifically bound to soluble antigen, without cross reacting with other irrelevant antigens. They also showed superior heat stability, exhibiting slow loss of activity over the course of one hour at high temperature (95 degrees C), while conventional antibodies lost all activity in the first 5-10min. The successful isolation of target specific sdAbs from one of our non-biased NAR libraries, demonstrate their ability to provide binders against an unacquainted antigen of interest.

Portal fusion protein constraints on function in DNA packaging of bacteriophage T4.

Architecturally conserved viral portal dodecamers are central to capsid assembly and DNA packaging. To examine bacteriophage T4 portal functions, we constructed, expressed and assembled portal gene 20 fusion proteins. C-terminally fused (gp20-GFP, gp20-HOC) and N-terminally fused (GFP-gp20 and HOC-gp20) portal fusion proteins assembled in vivo into active phage. Phage assembled C-terminal fusion proteins were inaccessible to trypsin whereas assembled N-terminal fusions were accessible to trypsin, consistent with locations inside and outside the capsid respectively. Both N- and C-terminal fusions required coassembly into portals with approximately 50% wild-type (WT) or near WT-sized 20am truncated portal proteins to yield active phage. Trypsin digestion of HOC-gp20 portal fusion phage showed comparable protection of the HOC and gp20 portions of the proteolysed HOC-gp20 fusion, suggesting both proteins occupy protected capsid positions, at both the portal and the proximal HOC capsid-binding sites. The external portal location of the HOC portion of the HOC-gp20 fusion phage was confirmed by anti-HOC immuno-gold labelling studies that showed a gold 'necklace' around the phage capsid portal. Analysis of HOC-gp20-containing proheads showed increased HOC protein protection from trypsin degradation only after prohead expansion, indicating incorporation of HOC-gp20 portal fusion protein to protective proximal HOC-binding sites following this maturation. These proheads also showed no DNA packaging defect in vitro as compared with WT. Retention of function of phage and prohead portals with bulky internal (C-terminal) and external (N-terminal) fusion protein extensions, particularly of apparently capsid tethered portals, challenges the portal rotation requirement of some hypothetical DNA packaging mechanisms.

Selective anterior fusion of thoracolumbar/lumbar curves in adolescents: when can the associated thoracic curve be left unfused?

STUDY DESIGN: A retrospective multicenter study was conducted to investigate patients with a major thoracolumbar/lumbar adolescent idiopathic scoliosis and an associated minor thoracic curve treated with an anterior instrumentation and fusion of the lower curve. OBJECTIVE: To establish criteria for determining when such curves can be successfully treated by an anterior only procedure of the lower curve with acceptable spinal balance and residual thoracic curve. SUMMARY OF BACKGROUND DATA: Anterior spinal instrumentation techniques have been proved effective for the management of isolated thoracolumbar/lumbar scoliosis with small compensatory thoracic curves. The success of a selective anterior fusion when the associated thoracic curve had some structural changes in a small preliminary study was variable and was the stimulus for this study. METHODS: A multicenter study involved 49 adolescent patients with a major thoracolumbar/lumbar curve in which the associated minor thoracic curve measured between 30 degrees and 55 degrees. In all the patients, the apical vertebra of the lower curve lay outside the midsacral line, and the thoracic apical vertebra fell outside a line dropped from the center of C7. Multiple radiographic parameters were evaluated. The Risser sign, height, weight, onset of menses, and closure of the triradiate cartilages were studied to access the patients' maturity. All the patients were observed at least 2 years. Patients were considered to have a satisfactory result if the thoracic curve at the final follow-up assessment measured 40 degrees or less, if balance and sagittal alignment were reasonable, and if additional procedures were not required. RESULTS: At final follow-up assessment, two groups emerged. Group 1 (n = 43) had satisfactory results. The preoperative thoracic curve in this group averaged 40 degrees and 26 degrees after surgery. The lumbar curve averaged 56 degrees before surgery and 22 degrees after surgery. Group 2 (n = 6) had unsatisfactory results. The average thoracic curve was 49 degrees before surgery 54 degrees after surgery, whereas the lumbar curve averaged 59 degrees before surgery and 27 degrees after surgery. Three of these patients underwent posterior thoracic instrumentation and fusion. CONCLUSIONS: Statistical analysis showed that a successful surgical outcome was dependent on both the structural changes in the thoracic curve and the patient's maturity. The thoracolumbar/lumbar-thoracic (TL/L:T) Cobb ratio in combination with the degree of the thoracic curve on lateral bending was the best predictor among the structural indexes. Of 44 patients with a TL/L:T Cobb ratio of 1.25 or greater and/or a thoracic curve, which bent out to 20 degrees or less, 42 had a satisfactory result. The best predictor among the maturity indexes was closure of the triradiate cartilages. Of 43 patients in whom the triradiate cartilages were closed, 42 had satisfactory results. When this data is combined, the outcome for the thoracic curve can be reasonably predicted.

Isolation and characterization of T4 bacteriophage gp17 terminase, a large subunit multimer with enhanced ATPase activity.

Phage T4 terminase is a two-subunit enzyme that binds to the prohead portal protein and cuts and packages a headful of concatameric DNA. To characterize the T4 terminase large subunit, gp17 (70 kDa), gene 17 was cloned and expressed as a chitin-binding fusion protein. Following cleavage and release of gp17 from chitin, two additional column steps completed purification. The purification yielded (i) homogeneous soluble gp17 highly active in in vitro DNA packaging ( approximately 10% efficiency, >10(8) phage/ml of extract); (ii) gp17 lacking endonuclease and contaminating protease activities; and (iii) a DNA-independent ATPase activity stimulated >100-fold by the terminase small subunit, gp16 (18 kDa), and modestly by portal gp20 and single-stranded binding protein gp32 multimers. Analyses revealed a preparation of highly active and slightly active gp17 forms, and the latter could be removed by immunoprecipitation using antiserum raised against a denatured form of the gp17 protein, leaving a terminase with the increased specific activity (approximately 400 ATPs/gp17 monomer/min) required for DNA packaging. Analysis of gp17 complexes separated from gp16 on glycerol gradients showed that a prolonged enhanced ATPase activity persisted after exposure to gp16, suggesting that constant interaction of the two proteins may not be required during packaging.

A bipartite bacteriophage T4 SOC and HOC randomized peptide display library: detection and analysis of phage T4 terminase (gp17) and late sigma factor (gp55) interaction.

HOC and SOC are dispensable T4 capsid proteins that can be used for phage display of multiple copies of peptides and proteins. A bipartite phage T4 peptide library was created by displaying on tetra-alanine linker peptides five randomized amino acids from the carboxyl-terminus of SOC and five randomized amino acids from the amino terminus of HOC. The bipartite library was biopanned against the phage T4 terminase large subunit gp17 to identify T4 gene products that may interact with the terminase. The sequences of selected phages displayed matches to those T4 gene products previously known by genetic and biochemical criteria to interact with gp17: gp20 (portal protein), gp32 (single-stranded DNA binding protein), gp16 (terminase small subunit), and gp17 (self). In addition, matches were found to gp55 (T4 late sigma factor), gp45 (sliding clamp), gp44 (clamp loader), gp2 (DNA end protein), and gp23 (major capsid protein). Abundant amino acid sequence matches were found to aa region 118-134 of gp55. Immunoprecipitation and affinity column chromatography demonstrated direct binding of gp17 and gp55; moreover, gp17 bound specifically to a column-coupled peptide corresponding to gp55 residues 111-136. Measurements of gene 17 and other mRNA levels in mutant-infected bacteria did not support a role of gp17-gp55 interaction in regulation of terminase or other late gene transcription. However, whereas DNA concatemers that accumulate in prohead and terminase defective phage T4 infections could be packaged in vitro to approximately 10% wild-type efficiency, 55am33am defective concatemeric DNA was packaged at least 100-fold less efficiently. Moreover, gp55 residues 111-136 peptide specifically blocked DNA packaging in vitro. These results suggest that the T4 terminase interaction with T4 late sigma factor gp55 plays a role in DNA packaging in vivo. The gp55 interaction may function to load the terminase onto DNA for packaging.