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This study investigates the use of low cost, customizable, biodegradable, polymer-ceramic composite porous structures (bone bricks) for large bone tissue regeneration. Different ceramic materials (hydroxyapatite (HA), β-tri-calcium phosphate (TCP) and Bioglass (45S5) were mixed with poly-ε-caprolactone (PCL). Bone bricks with different material compositions were produced using an extrusion-based additive manufacturing system. Produced bone bricks were morphologically and mechanically assessed. Results allowed to establish a correlation between scaffolds architecture and material composition and scaffolds performance. Reinforced scaffolds showed improved mechanical properties. Best mechanical properties were obtained with PCL/TCP bone bricks and topologies based on 38 double zig zag filaments and 14 spirals.

Nineteen patients receiving oxprenolol slow-release (SR) 160 mg (three patients) or 320 mg (16 patients) once daily for mild to moderate hypertension were treated with oxprenolol Oros 16/260 once daily for 3 weeks following a 2 week placebo wash-out period. Repeated dosing with both Oros and SR oxprenolol preparations, in comparison with placebo, significantly reduced supine systolic and diastolic blood pressures, and pulse rate at 24 h after dosing. Single Oros doses also significantly reduced pulse rate and diastolic, but not systolic, blood pressure at 24 h. The reduction in supine systolic blood pressure was greater during repeated dosing with oxprenolol SR than after a single dose of the Oros preparation. Control of supine diastolic blood pressure (less than or equal to 90 mm Hg) at 24 h after dosing was achieved in 13 out of 18 patients with oxprenolol SR (two out of three patients given 160 mg, and 11 out of 15 given 320 mg). Similar control was achieved in 11 out of 18 patients after a single dose of oxprenolol Oros, and in 13 out of 17 patients treated for 3 weeks. The mean percentage reduction in exercise heart rate (EHR) compared to placebo, at 24 h after dosing, was 16% following Oros treatment for 3 weeks, and 12% following SR administration. After a single dose of oxprenolol Oros EHR, was reduced by 9% at 24 h compared to placebo. At 3 weeks the Oros formulation was significantly better than the SR tablet at reducing EHR. Oxprenolol Oros 16/260 was effective over 24 h and well tolerated.(ABSTRACT TRUNCATED AT 250 WORDS) [ABSTRACT FROM AUTHOR]

Abstract Background The Atlantic salmon (Salmo salar) immunoglobulin heavy chain (IgH) locus possesses two parallel IgH isoloci (IGH-A and IGH-B), that are related to the genomic duplication event in the family Salmonidae. These duplicated IgH loci in Atlantic salmon provide a unique opportunity to examine the mechanisms of genome diversity and genome evolution of the IgH loci in vertebrates. In this study, we defined the structure of these loci in Atlantic salmon, and sequenced 24 bacterial artificial chromosome (BAC) clones that were assembled into the IGH-A (1.1 Mb) and IGH-B (0.9 Mb) loci. In addition, over 7,000 cDNA clones from the IgH variable (VH) region have been sequenced and analyzed. Results The present study shows that the genomic organization of the duplicated IgH loci in Atlantic salmon differs from that in other teleosts and other vertebrates. The loci possess multiple Cτ genes upstream of the Cμ region, with three of the Cτ genes being functional. Moreover, the duplicated loci possess over 300 VH segments which could be classified into 18 families. This is the largest number of VH families currently defined in any vertebrate. There were significant structural differences between the two loci, indicating that both IGH-A and -B loci have evolved independently in the short time after the recent genome duplication approximately 60 mya. Conclusions Our results indicate that the duplication of the IgH loci in Atlantic salmon significantly contributes to the increased diversity of the antibody repertoire, as compared with the single IgH locus in other vertebrates.

Abstract Background Two decades after the introduction of oil-based vaccines in the control of bacterial and viral diseases in farmed salmonids, the mechanisms of induced side effects manifested as intra-abdominal granulomas remain unresolved. Side effects have been associated with generation of auto-antibodies and autoimmunity but the underlying profile of inflammatory and immune response has not been characterized. This study was undertaken with the aim to elucidate the inflammatory and immune mechanisms of granuloma formation at gene expression level associated with high and low side effect (granuloma) indices. Groups of Atlantic salmon parr were injected intraperitoneally with oil-adjuvanted vaccines containing either high or low concentrations of Aeromonas salmonicida or Moritella viscosa antigens in order to induce polarized (severe and mild) granulomatous reactions. The established granulomatous reactions were confirmed by gross and histological methods at 3 months post vaccination when responses were known to have matured. The corresponding gene expression patterns in the head kidneys were profiled using salmonid cDNA microarrays followed by validation by real-time quantitative PCR (qPCR). qPCR was also used to examine the expression of additional genes known to be important in the adaptive immune response. Results Granulomatous lesions were observed in all vaccinated fish. The presence of severe granulomas was associated with a profile of up-regulation of innate immunity-related genes such as complement factors C1q and C6, mannose binding protein, lysozyme C, C-type lectin receptor, CD209, Cathepsin D, CD63, LECT-2, CC chemokine and metallothionein. In addition, TGF-β (p = 0.001), IL-17A (p = 0.007) and its receptor (IL-17AR) (p = 0.009) representing TH17 were significantly up-regulated in the group with severe granulomas as were arginase and IgM. None of the genes directly reflective of TH1 T cell lineage (IFN-γ, CD4) or TH2 (GATA-3) responses were differentially expressed. Conclusions Granulomatous reactions following vaccination with oil-based vaccines in Atlantic salmon have the profile of strong expression of genes related to innate immune responses. The expression of TGF-β, IL-17A and its receptor suggests an involvement of TH17 T cell lineage and is in conformity with strong infiltration of neutrophils and macrophages into inflamed areas. Arginase upregulation shows that macrophages in these reactions are alternatively activated, indicating also a TH2-profile. To what extent the expression of IL-17A and its receptor reflects an autoimmune vaccine-based reaction remains elusive but would be in conformity with previous observations of autoimmune reactions in salmon when vaccinated with oil-based vaccines.

Abstract Background Salmonids are one of the most intensely studied fish, in part due to their economic and environmental importance, and in part due to a recent whole genome duplication in the common ancestor of salmonids. This duplication greatly impacts species diversification, functional specialization, and adaptation. Extensive new genomic resources have recently become available for Atlantic salmon (Salmo salar), but documentation of allelic versus duplicate reference genes remains a major uncertainty in the complete characterization of its genome and its evolution. Results From existing expressed sequence tag (EST) resources and three new full-length cDNA libraries, 9,057 reference quality full-length gene insert clones were identified for Atlantic salmon. A further 1,365 reference full-length clones were annotated from 29,221 northern pike (Esox lucius) ESTs. Pairwise dN/dS comparisons within each of 408 sets of duplicated salmon genes using northern pike as a diploid out-group show asymmetric relaxation of selection on salmon duplicates. Conclusions 9,057 full-length reference genes were characterized in S. salar and can be used to identify alleles and gene family members. Comparisons of duplicated genes show that while purifying selection is the predominant force acting on both duplicates, consistent with retention of functionality in both copies, some relaxation of pressure on gene duplicates can be identified. In addition, there is evidence that evolution has acted asymmetrically on paralogs, allowing one of the pair to diverge at a faster rate.

Abstract Background Salmonids are of interest because of their relatively recent genome duplication, and their extensive use in wild fisheries and aquaculture. A comprehensive gene list and a comparison of genes in some of the different species provide valuable genomic information for one of the most widely studied groups of fish. Results 298,304 expressed sequence tags (ESTs) from Atlantic salmon (69% of the total), 11,664 chinook, 10,813 sockeye, 10,051 brook trout, 10,975 grayling, 8,630 lake whitefish, and 3,624 northern pike ESTs were obtained in this study and have been deposited into the public databases. Contigs were built and putative full-length Atlantic salmon clones have been identified. A database containing ESTs, assemblies, consensus sequences, open reading frames, gene predictions and putative annotation is available. The overall similarity between Atlantic salmon ESTs and those of rainbow trout, chinook, sockeye, brook trout, grayling, lake whitefish, northern pike and rainbow smelt is 93.4, 94.2, 94.6, 94.4, 92.5, 91.7, 89.6, and 86.2% respectively. An analysis of 78 transcript sets show Salmo as a sister group to Oncorhynchus and Salvelinus within Salmoninae, and Thymallinae as a sister group to Salmoninae and Coregoninae within Salmonidae. Extensive gene duplication is consistent with a genome duplication in the common ancestor of salmonids. Using all of the available EST data, a new expanded salmonid cDNA microarray of 32,000 features was created. Cross-species hybridizations to this cDNA microarray indicate that this resource will be useful for studies of all 68 salmonid species. Conclusion An extensive collection and analysis of salmonid RNA putative transcripts indicate that Pacific salmon, Atlantic salmon and charr are 94–96% similar while the more distant whitefish, grayling, pike and smelt are 93, 92, 89 and 86% similar to salmon. The salmonid transcriptome reveals a complex history of gene duplication that is consistent with an ancestral salmonid genome duplication hypothesis. Genome resources, including a new 32 K microarray, provide valuable new tools to study salmonids.

Abstract Background We have previously identified associations between major histocompatibility complex (MHC) class I and resistance towards bacterial and viral pathogens in Atlantic salmon. To evaluate if only MHC or also closely linked genes contributed to the observed resistance we ventured into sequencing of the duplicated MHC class I regions of Atlantic salmon. Results Nine BACs covering more than 500 kb of the two duplicated MHC class I regions of Atlantic salmon were sequenced and the gene organizations characterized. Both regions contained the proteasome components PSMB8, PSMB9, PSMB9-like and PSMB10 in addition to the transporter for antigen processing TAP2, as well as genes for KIFC1, ZBTB22, DAXX, TAPBP, BRD2, COL11A2, RXRB and SLC39A7. The IA region contained the recently reported MHC class I Sasa-ULA locus residing approximately 50 kb upstream of the major Sasa-UBA locus. The duplicated class IB region contained an MHC class I locus resembling the rainbow trout UCA locus, but although transcribed it was a pseudogene. No other MHC class I-like genes were detected in the two duplicated regions. Two allelic BACs spanning the UBA locus had 99.2% identity over 125 kb, while the IA region showed 82.5% identity over 136 kb to the IB region. The Atlantic salmon IB region had an insert of 220 kb in comparison to the IA region containing three chitin synthase genes. Conclusion We have characterized the gene organization of more than 500 kb of the two duplicated MHC class I regions in Atlantic salmon. Although Atlantic salmon and rainbow trout are closely related, the gene organization of their IB region has undergone extensive gene rearrangements. The Atlantic salmon has only one class I UCA pseudogene in the IB region while trout contains the four MHC UCA, UDA, UEA and UFA class I loci. The large differences in gene content and most likely function of the salmon and trout class IB region clearly argues that sequencing of salmon will not necessarily provide information relevant for trout and vice versa.

Abstract Background We have developed and fabricated a salmonid microarray containing cDNAs representing 16,006 genes. The genes spotted on the array have been stringently selected from Atlantic salmon and rainbow trout expressed sequence tag (EST) databases. The EST databases presently contain over 300,000 sequences from over 175 salmonid cDNA libraries derived from a wide variety of tissues and different developmental stages. In order to evaluate the utility of the microarray, a number of hybridization techniques and screening methods have been developed and tested. Results We have analyzed and evaluated the utility of a microarray containing 16,006 (16K) salmonid cDNAs in a variety of potential experimental settings. We quantified the amount of transcriptome binding that occurred in cross-species, organ complexity and intraspecific variation hybridization studies. We also developed a methodology to rapidly identify and confirm the contents of a bacterial artificial chromosome (BAC) library containing Atlantic salmon genomic DNA. Conclusion We validate and demonstrate the usefulness of the 16K microarray over a wide range of teleosts, even for transcriptome targets from species distantly related to salmonids. We show the potential of the use of the microarray in a variety of experimental settings through hybridization studies that examine the binding of targets derived from different organs and tissues. Intraspecific variation in transcriptome expression is evaluated and discussed. Finally, BAC hybridizations are demonstrated as a rapid and accurate means to identify gene content.

Background: To compare relative rates of pediatric poisoning before and after COVID-19, including by demographic and urban-rural status, and by agent identified, using data from one university healthcare system and children's hospital., Methods: Using retrospective, cross sectional design from deidentified healthcare claims data, we extracted all encounters with the ICD-10-CM for Poisoning by, Adverse effects of, and Underdosing of drugs, medicants and biological substances (T36-T50) and grouped the encounters as those after state mandates regulating activity came into effect (Post-COVID-19 (3/17/2020-3/18/2021)) Pre-COVID-19 (3/18/2019-3/17/2020). We then compared poisoning agent, age at the time of the encounter, recorded sex, race, ethnicity, rural/urban residence, and visit type using Mann-Whitney U test, chi-square test of association, incidence rates and incident rate ratios between the time periods., Findings: The sample included 1608 unique patients 0-17 years of age and 4216 encounters. We also identified IRRs >1 in nearly every demographic subgroup with the exception of Non-Hispanic Blacks. The comparison of specific drugs or medicants identified a significant decrease in poisoning by Systemic antibiotics (T36); but an increase in Hormones and their synthetic substitutes and antagonists (T38), Non opioid analgesics antipyretic and antirheumatic (T39), Psychotropic Drugs (T39) and Systemic and hematologic agents (T45)., Conclusion: This study identifies pediatric subgroups highly affected by pediatric poisoning during the time-period immediately after the identification of COVID-19 and characterizes the drugs commonly associated with poisonings., Application to Practice: With a further understanding nursing has the potential to impact pediatric poisoning in the inpatient, outpatient and public health setting., Competing Interests: Declaration of competing interest Authors do not have any interests to report related to this work., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Purpose: Those factors identified to increase the risk of suicide in rural dwellers were exacerbated by the SARS-CoV-2 pandemic, specifically economic factors, substance use, access to health care, and access to lethal weapons. Because the effects of SARS-CoV-2 on suicide ideation and attempts in rural populations have not been fully characterized in published literature, this study compares: (1) the rates of suicide ideation and attempts between the 6 months affected by SARS-CoV-2 to same months of the preceding year (3/18/2020-9/18/20; 3/18/2019-9/18/19), (2) demographics (ie, age, sex, residence, race, and ethnicity), and (3) the locations in which the encounters were billed (inpatient, outpatient, and emergency department)., Methods: Deidentified claims data associated with patient encounters billed for Suicide Ideation and Suicide Attempt were grouped based on time period and analyzed using descriptive statistics, incidence rate ratio (IRR), 2-sample t-test, chi-square test of association, or Fisher's exact test., Findings: Suicidal ideation encounters increased in the 6 months post-SARS-CoV-2 when compared to the 6 months of the prior year (IRR = 1.19; P < .001). Males (IRR = 1.27, P < .001), those residing rural areas (IRR = 1.22, P = .01), and Black, non-Hispanic (IRR = 1.24, P = .024) were found to have increased rates of suicide ideation post-SARS-Cov-2. In adults, White, non-Hispanics (IRR = 1.16; P < .001) had increased rates of post-SARS-CoV-2. In the pediatric subset, those who were aged 14-17 (IRR = 1.50; P < .001), resided in rural areas (IRR = 1.61, P = .009), and idenitifed as Hispanic (IRR = 1.89; P = .037) or Black, non-Hispanic (IRR = 1.61, P = .009) had increased rates post-SARS-CoV-2., Conclusions: Our study identified rural dwellers to be at increased risk for suicide ideation., (© 2022 National Rural Health Association.)

Background: Risk factors for child abuse and neglect and commonly used reporting mechanisms were highly affected by SARS-Cov-2 pandemic; yet, little is known about the effects of SARS-Cov-2 on rates of child abuse and neglect., Objective: To compare overall rates, demographics, types of abuse and acuity of child abuse and neglect encounters seen at one university health system for the 6 months before and after school closings due to the SARS-Cov-2 pandemic., Participants and Setting: Data was extracted from a database of billed ICD10 codes for child abuse and neglect including sexual abuse codes. There were 579 encounters for patients <18 years of age and 476 unique patients., Methods: In addition to ICD10 code and pre/post school closing, each encounter was identified to be inpatient, outpatient and/or emergency department. Demographic data such as age, gender, ethnicity, and race were extracted. Incident rate ratios in addition to descriptive statistics, Mann-Whitney U test, two-sample t-test, or the chi-square test of association were used in the analysis., Results: No significant differences were identified for total rates of child abuse and neglect encounters (p = .08), physical abuse (p = .91) nor child maltreatment (p = .86) codes or in the age (p = .46), gender (p = .58), and race/ethnicity (p = .15) of patient encounters pre- versus post-school closings. The sexual abuse incidence and inpatient encounters increased by 85% (IRR = 1.85, p < .0001; IRR = 1.85, p = .004, respectively)., Conclusions: Our findings provide a unique contribution to the existing literature in that we identified a significant increase in the incidence of sexual abuse and higher patient acuity as evidenced by higher rates of inpatient encounters after school closing due to SARS-Cov-2., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Context: Previous research from a sample of US secondary schools (n = 10 553) indicated that 67% of schools had access to an athletic trainer (AT; 35% full time [FT], 30% part time [PT], and 2% per diem). However, the population-based statistic in all secondary schools with athletic programs (n = approximately 20 000) is yet to be determined., Objective: To determine the level of AT services and employment status in US secondary schools with athletics by National Athletic Trainers' Association district., Design: Cross-sectional study., Setting: Public and private secondary schools with athletics., Patients or Other Participants: Data from all 20 272 US public and private secondary schools were obtained., Main Outcome Measure(s): Data were collected from September 2015 to April 2018 by phone or e-mail communication with school administrators or ATs and by online surveys of secondary school ATs. Employment categories were school district, school district with teaching, medical or university facility, and independent contractor. Data are presented as total number and percentage of ATs. Descriptive statistics were calculated for FT, PT, and no AT services data for public, private, public + private, and employment type by state and by National Athletic Trainers' Association district., Results: Of the 20 272 secondary schools, 66% (n = 13 473) had access to AT services, while 34% (n = 6799) had no access. Of those schools with AT services, 53% (n = 7119) received FT services, while 47% (n = 6354) received PT services. Public schools (n = 16 076) received 37%, 32%, and 31%, whereas private schools (n = 4196) received 27%, 28%, and 45%, for FT, PT, and no AT services, respectively. Most of the Athletic Training Locations and Services Survey participants (n = 6754, 57%) were employed by a medical or university facility, followed by a school district, school district with teaching, and independent contractor. Combined, 38% of AT employment was via the school district., Conclusions: The percentages of US schools with AT access and FT and PT services were similar to those noted in previous research. One-third of secondary schools had no access to AT services. The majority of AT employment was via medical or university facilities. These data depict the largest and most updated representation of AT services in secondary schools.

This study describes the molecular characterization of avian reoviruses (ARVs) isolated during an outbreak in commercial chickens between 2015 and 2016. In addition, a pathogenicity study of a selected ARV strain isolated from a field case of viral tenosynovitis in commercial broiler chickens was performed. On the basis of phylogenetic analysis of a 1088-bp fragment of the ARV S1 gene, the investigated sequences were differentiated into five distinct genotypic clusters (GCs), namely GC1, GC2, GC3, GC4, and GC6. Specific-pathogen-free (SPF) and commercial broiler chickens were challenged with the GC1 genetic type MK247011, at 14 days of age via the interdigital toe web. No significant effects in body weight gain and feed conversion were detected in both chicken types. The Δ interdigital web thickness was most severe at 4 days postchallenge (DPC) in both the SPF and broiler subgroups. The inflammation in SPF birds was slightly more severe compared with broilers. Neither mortality nor clinical signs occurred in the infected groups for the duration of the experiment, despite the presence of significant microscopic lesions in challenged birds. Microscopic changes of tenosynovitis became evident at 3 DPC, with the highest incidence and severity detected at 14 and 21 DPC, respectively. Seroconversion against ARV occurred 3 wk postchallenge, and the microscopic lesions detected in tendon and heart sections were highly compatible with those described in the field. Increased severity of tenosynovitis and epicarditis lesions were noted in the ARV-challenged groups compared with the control groups. Although SPF and broiler chickens showed comparable responses to the challenge with an ARV genetic variant, detected lesions were subclinical, denoting the limitations of our challenge approach. The age selected in this experiment possibly influenced the course of the infection. Data from this study highlight the genotypic diversity of isolates in California, and the outcome of the pathogenicity study can be used as a basis to improve protocols for pathogenicity studies to characterize ARV variants causing clinical disease in the field.

Tau is a microtubule-associated protein that normally interacts in monomeric form with the neuronal cytoskeleton. In Alzheimer's disease, however, it aggregates to form the structural component of neurofibrillary lesions. The transformation is controlled in part by age- and disease-associated post-translational modifications. Recently we reported that tau isolated from cognitively normal human brain was methylated on lysine residues, and that high-stoichiometry methylation depressed tau aggregation propensity in vitro. However, whether methylation stoichiometry reached levels needed to influence aggregation propensity in human brain was unknown. Here we address this problem using liquid chromatography-tandem mass spectrometry approaches and human-derived tau samples. Results revealed that lysine methylation was present in soluble tau isolated from cognitively normal elderly cases at multiple sites that only partially overlapped with the distributions reported for cognitively normal middle aged and AD cohorts, and that the quality of methylation shifted from predominantly dimethyl-lysine to monomethyl-lysine with aging and disease. However, bulk mol methylation/mol tau stoichiometries never exceeded 1 mol methyl group/mol tau protein. We conclude that lysine methylation is a physiological post-translational modification of tau protein that changes qualitatively with aging and disease, and that pharmacological elevation of tau methylation may provide a means for protecting against pathological tau aggregation.

Infectious coryza, caused by Avibacterium paragallinarum, is an acute respiratory disease of poultry that can result in substantial morbidity, mortality, and economic losses. In March 2017, the Turlock branch of the California Animal Health and Food Safety laboratory system encountered an unusual clinical and pathologic presentation of infectious coryza in 6 live, 29-d-old, commercial broiler chickens that were submitted for diagnostic investigation. Antemortem evaluation revealed severe neurologic signs, including disorientation, torticollis, and opisthotonos. Swollen head-like syndrome and sinusitis were also present. Histologically, severe sinusitis, cranial osteomyelitis, otitis media and interna, and meningoencephalitis were noted, explaining the clinical signs described. A. paragallinarum was readily isolated from the upper and lower respiratory tract, brain, and cranial bones. Infectious bronchitis virus (IBV) was also detected by PCR, and IBV was isolated in embryonated chicken eggs. Based on sequencing analysis, the IBV appeared 99% homologous to strain CA1737. A synergistic effect between A. paragallinarum and IBV, resulting in exacerbation of clinical signs and increased mortality, may have occurred in this case. A. paragallinarum should be considered among the possible causes of neurologic signs in chickens. Appropriate media should be used for bacterial isolation, and the role of additional contributing factors and/or complicating agents should be investigated in cases of infectious coryza.

Streptococcal bacterial species represent common inhabitants of the intestinal tract of animals and humans with a potential for opportunistic infections. Streptococcosis has been identified in turkey poults ( Meleagris gallopavo), ducklings and goslings (Anatidae), broiler chickens, semimature-adult chickens ( Gallus gallus domesticus), and young and adult pigeons (Columbidae). However, the exact underlying factors that lead to bacterial invasion of the blood stream and tissue colonization have not been completely elucidated. The electronic database of the California Animal Health and Food Safety laboratory (Fresno, Tulare, and Turlock branches) was searched for necropsy cases in which streptococcosis was diagnosed in different avian species between January 2000 and August 2017. A total of 95 cases, involving both commercial operations and noncommercial premises, were analyzed. Streptococcus spp., Streptococcus bovis, and Streptococcus gallolyticus were identified from multiple organs, with macroscopic or histopathologic lesions (or both) indicative of septicemia in 23 (24%), 40 (42%), and 30 (32%) cases, respectively. Streptococcus pluranimalium and Streptococcus lutetiensis were also isolated from one (1%) and two (2%) cases, respectively. Turkey poults, broiler chickens, and ducklings were the most-commonly affected species with streptococcosis. Splenitis and hepatitis were the most-common lesions observed and these were the organs with the highest isolation rate. An overview of the clinical and pathologic presentation, and possible predisposing conditions associated with this bacterial infection, is provided.

Post-translational modifications are biologically important and wide-spread modulators of protein function. Although methods for detecting the presence of specific modifications are becoming established, approaches for quantifying their mol modification/mol protein stoichiometry are less well developed. Here we introduce a ratiometric, label-free, targeted liquid chromatography tandem mass spectroscopy-based method for estimating Lys and Arg methylation stoichiometry on post-translationally modified proteins. Methylated Lys and Arg were detected with limits of quantification at low fmol and with linearity extending from 20 to 5000 fmol. This level of sensitivity allowed estimation of methylation stoichiometry from microgram quantities of various proteins, including those derived from either recombinant or tissue sources. The method also disaggregated total methylation stoichiometry into its elementary mono-, di-, and tri-methylated residue components. In addition to being compatible with kinetic experiments of protein methylation, the approach will be especially useful for characterizing methylation states of proteins isolated from cells and tissues., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In February 2015, two Eurasian collared doves (Streptopelia decaocto) were submitted dead to the California Animal Health and Food Safety (CAHFS) Laboratory, Turlock branch, from a private aviary experiencing sudden, high mortality (4/9) in adult doves. In both doves, the gross and histologic lesions were indicative of acute, fatal septicemia. Grossly, there were numerous pale yellow foci, 1 to 2 mm in diameter, in the liver and spleen. Microscopically, these foci were composed of acute severe multifocal coagulative necrosis of hepatocytes and splenic pulp with infiltration of heterophils mixed with fibrin and dense colonies of gram-negative bacteria. Yersinia pseudotuberculosis was isolated from the lung, liver, spleen, heart, ovary, kidney, and trachea. The organism was susceptible to most antibiotics it was tested against, except erythromycin. Based on a retrospective study of necropsy submissions to CAHFS between 1990 and 2015, there were 77 avian case submissions of Y. pseudotuberculosis. There were 75/77 cases identified from a wide range of captive avian species from both zoo and private facilities and 2/77 cases from two backyard turkeys submitted from one premise. The largest number of cases originated from psittacine species (31/77). The lesions most commonly described were hepatitis (63/77), splenitis (49/77), pneumonia (30/77), nephritis (16/77), and enteritis (12/77). From 1990 to 2015, there was an average of three cases of avian pseudotuberculosis per year at CAHFS. Although there were no cases diagnosed in 1993 and 1994, in all other years, there were between one and eight cases of Y. pseudotuberculosis detected from avian diagnostic submissions.

In Alzheimer's disease, the microtubule-associated protein tau dissociates from the neuronal cytoskeleton and aggregates to form cytoplasmic inclusions. Although hyperphosphorylation of tau serine and threonine residues is an established trigger of tau misfunction and aggregation, tau modifications extend to lysine residues as well, raising the possibility that different modification signatures depress or promote aggregation propensity depending on site occupancy. To identify lysine residue modifications associated with normal tau function, soluble tau proteins isolated from four cognitively normal human brains were characterized by MS methods. The major detectable lysine modification was found to be methylation, which appeared in the form of mono- and di-methyl lysine residues distributed among at least 11 sites. Unlike tau phosphorylation sites, the frequency of lysine methylation was highest in the microtubule-binding repeat region that mediates both microtubule binding and homotypic interactions. When purified recombinant human tau was modified in vitro through reductive methylation, its ability to promote tubulin polymerization was retained, whereas its aggregation propensity was greatly attenuated at both nucleation and extension steps. These data establish lysine methylation as part of the normal tau post-translational modification signature in human brain, and suggest that it can function in part to protect against pathological tau aggregation.

Since the discovery of phenothiazines as tau protein aggregation inhibitors, many additional small molecule inhibitors of diverse chemotype have been discovered and characterized in biological model systems. Although direct inhibition of tau aggregation has shown promise as a potential treatment strategy for depressing neurofibrillary lesion formation in Alzheimer's disease, the mechanism of action of these compounds has been unclear. However, recent studies have found that tau aggregation antagonists exert their effects through both covalent and non-covalent means, and have identified associated potency and selectivity driving features. Here we review small-molecule tau aggregation inhibitors with a focus on compound structure and inhibitory mechanism. The elucidation of inhibitory mechanism has implications for maximizing on-target efficacy while minimizing off-target side effects.

Small molecules that bind tau-bearing neurofibrillary lesions are being sought for premortem diagnosis, staging, and treatment of Alzheimer's disease and other tauopathic neurodegenerative diseases. The utility of these agents will depend on both their binding affinity and binding site density (B(max)). Previously we identified polarizability as a descriptor of protein aggregate binding affinity. To examine its contribution to binding site density, we investigated the ability of two closely related benzothiazole derivatives ((E)-2-[[4-(dimethylamino)phenyl]azo]-6-methoxybenzothiazole) and ((E)-2-[2-[4-(dimethylamino)phenyl]ethenyl]-6-methoxybenzothiazole) that differed in polarizability to displace probes of high (Thioflavin S) and low (radiolabeled (E,E)-1-iodo-2,5-bis(3-hydroxycarbonyl-4-methoxy)styrylbenzene; IMSB) density sites. Consistent with their site densities, Thioflavin S completely displaced radiolabeled IMSB, but IMSB was incapable of displacing Thioflavin S. Although both benzothiazoles displaced the low B(max) IMSB probe, only the highly polarizable analog displaced near saturating concentrations of the Thioflavin S probe. Quantum calculations showed that high polarizability reflected extensive pi-electron delocalization fostered by the presence of electron donating and accepting groups. These data suggest that electron delocalization promotes ligand binding at a subset of sites on tau aggregates that are present at high density, and that optimizing this aspect of ligand structure can yield tau-directed agents with superior diagnostic and therapeutic performance., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This report documents the occurrence of a very virulent infectious bursal disease virus (vvIBDV) in Northern California commercial brown pullets. Diagnosis was made from multiple accessions from two neighboring and epidemiologically related ranches submitted to the California Animal Health and Food Safety (CAHFS) laboratory. Pullets, 11 and 14 wk of age from ranch A (rA) and ranch B (rB) respectively, were submitted from infectious bursal disease virus vaccinated flocks experiencing a drastic increase in mortality. The December 2008 outbreak resulted in 26% and 34% mortality on rA and rB respectively. Gross and histologic lesions characteristic of acute vvIBDV were observed. Gross lesions included edematous bursas, hemorrhages at the junction of the proventriculus and gizzard as well as hemorrhages on skeletal muscles. Microscopic lesions included severe lymphoid necrosis and inflammation in edematous bursas, lymphoid necrosis in thymus, spleen, Peyer's patches and cecal tonsils. Diagnosis of vvIBDV was confirmed by molecular characterization of the IBDV from bursas as well as viral pathogenicity in specific-pathogen-free birds. RT-PCR and nucleotide sequencing of the hypervariable region of the VP2 (vVP2) gene segment of the IBDV genome was performed on rA, rB and embryo passaged rA virions.The amino acids compatible with vvIBDV isolates: 222(Ala), 242(Ile), 256(Ile), 294(Ile) and 299(Ser) were reported from both ranches. In addition, nucleotide sequencing of a fragment of the VP1 gene demonstrated the viruses have the segment B genotype associated with highly pathogenic vvIBDV. Inocula of 10(5.5) 50% egg infective dose of vvIBDV virus from rA and rB were introduced orally into two groups (g1 and g2 respectively) of 4 wk 2-day-old SPF leghorns. At 4 days postinoculation, there was 100% (22/22) morbidity in g1 and g2; 91% (20/22) mortality in g1; 100% (22/22) mortality for g2; 0% (0/20) morbidity and 0% (0/ 20) mortality was reported in the control group. This is the first occurrence of vvIBDV reported from birds in the United States.

Objectives: Eye injuries are relatively common following detonations. The majority of these are caused by secondary fragments. Primary blast injury to the eye (from the air-blast alone) has been described historically, but the differentiation of air-blast and fragment aetiologies in the reports is not clear. The aim of this review is to consider the evidence in the literature for true primary blast injury to the eye., Methods: A literature review using Medline, Pub-med and Web of Knowledge databases., Conclusions: Overall, there is little conclusive evidence that primary ocular blast injury occurs in survivors of explosions. However, some case reports do surmise its occurrence and it cannot be unequivocally ruled out. If it does occur, it is more likely to be at high overpressures that will produce life-threatening injuries. The development of enhanced blast weapons may result in an increased incidence. At present however, the most severe and damaging threat to the eyes from blast remains the impact of fragments and debris.

An outbreak of infectious catarrhal enteritis, associated with the flagellated protozoan Spironucleus meleagridis (syn. Hexamita meleagridis), is reported from a commercial flock of chukar partridges in California. The disease affected birds between the ages of 4 and 6 wk and resulted in diarrhea, listlessness, depression, and high mortality. Concurrent infection with other intestinal pathogens, including Cryptosporidia, group E Salmonella, long-segmented filamentous microorganisms (LSFMOs), and Rotavirus-like virus particles, was found in some but not all affected birds. Dermatitis of the face, shanks, and feet, suggestive of B-complex vitamin deficiency, was present in most affected birds as well. Flagellated protozoan parasites could be found in the lumen of the duodenum and jejunum and in the intestinal crypts. In some cases the flagellates were wedged between epithelial cells or were located intracellularly within cells of the mucosal epithelium and the intestinal lamina propria.

A disease characterized by paresis and paralysis was seen in 7-9-day-old broiler chicks after vaccination in the neck area at day-of-age with a live virus vaccine containing viruses of Marek's disease, fowl pox, and infectious bursal disease. Affected birds presented with variable signs of ataxia, lateral recumbency, leg paralysis, and twisting or S-shaped flexure of the neck. Gross lesions noted at necropsy included swelling and edema of the subcutaneous tissues and muscles of the neck at the injection site area. A heavy mononuclear inflammatory cell infiltration was seen in the subcutaneous tissues, connective tissues, and muscles of the neck at the injection site. In some cases, the inflammatory process extended along fascial planes to involve the epidural spaces surrounding the spinal cord. Fatty changes with possible demyelination of nerve fibers were noted in some sections of the spinal cord adjacent to the inflammatory lesions. Clusters of poxviruses were found within some inflammatory lesions on transmission electron photomicrographs.

Systemic erysipelas infection was seen in 2-to-4-day-old poults from three separate ranches owned by the same company. The affected poults were all from the same breeder source; poults from other breeder sources were seemingly unaffected. Mortality increased on days 2 and 3, ranging from 2% to 8.5%. Birds submitted were dehydrated and very weak, with one half of the poults submitted having died during transport to the lab. Gross lesions included swollen, congested livers and spleens, as well as hemorrhagic breast muscle in one case. Toes were swollen and reddish-purple in color. The poults had been toe-trimmed during hatchery processing using a commercial microwave. Histologically, periportal inflammation with heterophilic infiltration in the liver was noted. Spleens showed hyalinization of arteries, lymphoid depletion, and necrosis. Toe joints showed purulent synovitis and cellulitis. Gram stains done on impression smears of liver and spleen showed rare to moderate numbers of small gram-positive rods. Erysipelothrix rhusiopathiae was isolated from 18 of 22 livers cultured, five of six toe joints cultured, and from the yolk sac in two birds.

Egg production drops associated with western equine encephalitis (WEE) virus infection occurred in three turkey breeder flocks in California during summer 1993 and again in one flock the following year. Egg production losses totaled 8.76%, 9.57%, 9.71%, and 10.12% and were accompanied by an increase in small white-shelled and shell-less eggs. The outbreaks coincided with peak WEE virus activity in the state on the basis of statistics compiled by the California Department of Health Services on seroconversion rates in sentinel chicken flocks. Paired serum samples taken 2-3 wk apart showed increased titers to WEE between acute and convalescent sera in turkeys from three affected flocks. Convalescent sera were not available for testing from the fourth flock. WEE virus was isolated from one breeder hen submitted to the diagnostic laboratory during the early stages of the outbreak.

Seven 18-wk-old pullets from a commercial layer flock experiencing increased mortality associated with neurologic and respiratory symptoms were submitted to the California Veterinary Diagnostic Laboratory System at the Turlock Branch for necropsy. Clinical signs included depression, torticollis, swollen eyelids, conjunctivitis, and sinusitis. Meningoencephalitis and suppurative inflammation of the cranial air spaces were found on histopathology. The brain, sinuses, and air spaces of the cranium were infected with Pasteurella multocida. Complicating the condition was Mycoplasma gallisepticum infecting the sinus and paramyxovirus-I affecting the trachea.

A mixed breed rooster, from a backyard flock of 13 chickens, was received at California Veterinary Diagnostic Laboratory System-Turlock Branch for postmortem examination. The bird presented with thickened, featherless, scab-encrusted skin around the head region. Numerous sticktight fleas were found attached to the encrusted skin. Microscopic evaluation of the skin revealed a lymphoplasmacytic reaction in the dermis with visible embedded flea mouthparts. Also noted histologically in this region were epidermal hyperplasia and ballooned epidermal cells containing intracytoplasmic inclusions indicative of fowl poxvirus.

The pathogenicity of a field isolate of western equine encephalitis (WEE) virus, which was recovered from a breeder hen during investigations of egg production drops in California turkey flocks, was tested in 2-wk-old turkey poults. No symptoms or mortality were observed in poults inoculated intramuscularly with 4.2 log10 50% embryo lethal doses of virus; however, the infection did result in mild to moderate lymphoid necrosis in the bursa of Fabricius and thymus glands beginning on the first day postinoculation. In addition, WEE virus could be isolated from the blood of infected poults for up to 3 days postinoculation.

The serological responses to Salmonella enteritidis flagella (H: g,m) and its fimbrial antigen SEF14 were evaluated as indicators of infection in chickens and to confirm serological results obtained by an ELISA using S enteritidis lipopolysaccharide (LPS) (O: 9,12) as the detecting antigen. The SEF14 antigen and flagella were extracted from S enteritidis and transferred to nitrocellulose paper for use in Western and dot blot tests. Antisera to 19 salmonella serotypes including S enteritidis were raised in rabbits and their cross reactivity to the flagellar and SEF14 antigens was evaluated. Cross reactivity with the SEF14 antigen was found in one antiserum, raised against S blegdam, and to flagella in eight of 19 antisera raised against various salmonella serotypes, most of which shared the flagellar factors g or m with S enteritidis. The intensity of cross reaction to flagella was strongest in S derby and S blegdam antisera. Antisera raised in chickens against S typhimurium and S panama did not cross react in either test, and neither did pooled sera from eight-week-old salmonella-free, broiler breeder parent chickens. Field sera from two commercial flocks with no history of salmonella infection were negative when tested by the LPS ELISA. These sera were also negative when tested by the flagellar and SEF14 blots. S enteritidis infection in a commercial laying flock was detected initially when the sera were tested by the LPS ELISA and confirmed in individual and pooled sera by the SEF14 and flagellar tests. S enteritidis PT4 was isolated from this flock post mortem.

A meat turkey ranch had a history of producing flocks with low market weights. Live bird samples and environmental samples from the ranch were collected over a period of eight months and analyzed at the California Veterinary Diagnostic Laboratory System-Turlock Branch. The disease condition consistently present was enteritis associated with Ascaridia dissimilis infection in birds from growout houses. After aggressive control measures for A. dissimilis were implemented, subsequent flocks reached expected market weights.

Esophageal trichomoniasis has been rarely reported in chickens. At the California Veterinary Diagnostic Laboratory System-Turlock Branch, this disease was recently diagnosed in two cases submitted from backyard chicken flocks. The esophageal lesions observed were similar to those seen in several other important diseases of chickens. The causative trichomonad organisms were readily demonstrated on wet smears and by histologic studies. In both cases, the investigated flocks were afflicted with several concurrent diseases. California has experienced an increase in the number of small nontraditional chicken production operations. These facilities are sometimes in close proximity to commercial poultry operations and biosecurity barriers occasionally fail. The poor husbandry practices often used in these small flocks make them a potential reservoir for rare diseases such as trichomoniasis and also for disease organisms that are devastating to commercial poultry.

Outbreaks of enteritis associated with an unusual flagellated protozoan occurred in six California turkey flocks during the summer of 1992. Certain morphological and ultrastructural details of the parasite, which resembles Cochlosoma anatis, are illustrated with scanning and transmission electron micrographs. The flagellate attached to the intestinal mucosa by means of a sucker-like apparatus, and circular impressions of the sucker were created on the surface epithelium. Histological lesions were characterized by blunting and fusion of villi; cellular infiltration of the lamina propria with lymphocytes, plasma cells, histiocytes, and heterophils; and increased numbers of mitotic figures in crypt epithelium.

Newly hatched chicks were vaccinated orally with a genetically defined Salmonella enteritidis aroA candidate, strain CVL30. In chickens immunized with 10(5) or 10(9) CFU and challenged by the intravenous route with 10(8) CFU of S. enteritidis 109 Nalr at 8 weeks old, there were similar reductions in colonization of the spleens, livers, and ceca of vaccinees compared with unvaccinated controls. Two groups of newly hatched female chicks were vaccinated orally with 10(9) CFU of strain CVL30, and one group was revaccinated intramuscularly with 10(9) CFU at 16 weeks old. When challenged intravenously with S. enteritidis 109 Nalr at 23 weeks old, there was a reduction in the colonization of spleens, livers, ovaries, and ceca compared with unvaccinated controls. Inclusion of the intramuscular booster gave increased protection to the ovary, although the vaccine strain was isolated on one occasion from a batch of eggs laid at 20 weeks old. In chickens immunized with 10(9) CFU of strain CVL30 and challenged orally with 10(9) CFU of S. enteritidis 109 Nalr, there was a reduction in intestinal shedding of the challenge strain from vaccines compared with unvaccinated controls. Circulating immunoglobulin G antibodies to lipopolysaccharide (LPS) were detected in unvaccinated controls within 7 to 10 days of oral challenge. In contrast, circulating immunoglobulin G antibodies to LPS in vaccinees were not altered by the oral challenge, which suggested that vaccination reduced or prevented invasion by the challenge strain from the gut or multiplication of the challenge strain in the tissues. Newly hatched chicks were vaccinated orally with ca. 10(9) CFU of strain CVL30, and 1 day later, the vaccines and unvaccinated controls were challenged orally with 10(5) or 10(9) CFU of S. enteritidis 109 Nalr. Colonization of the ceca and invasion from the gut by the S. enteritidis challenge strain was reduced in the vaccines up to 5 days postchallenge compared with controls. In a second trial, vaccinees and controls were challenged orally with 10(7) or 10(9) CFU of S. typhimurium 2391 Nalr. In contrast to the challenge with S. enteritidis, colonization of the ceca and invasion by the S. typhimurium strain were not greatly reduced.

Newly hatched chicks were dosed orally with a Salmonella typhimurium wild-type strain, an S. enteritidis wild-type strain, and a genetically defined S. enteritidis aroA vaccine candidate, strain CVL30. The S. typhimurium strain, 2391 Nalr, was virulent in newly hatched chicks and caused deaths in 7 of 20 chicks after an oral dose of 10(5) organisms. The S. enteritidis wild-type strain, LA5, caused death in 1 of 25 chicks and gross pathology including pericarditis and perihepatitis in 6 of the 24 survivors after an oral dose of 10(9) organisms. S. enteritidis aroA CVL30, attenuated by ca. 6.5 log10 in BALB/c mice, was nonvirulent when administered orally to chicks and did not cause morbidity. When newly hatched chicks were dosed, the pattern of invasion and colonization of the reticuloendothelial system by strain CVL30 was similar to that of its parent strain, LA5, irrespective of the dose. Oral inoculation of newly hatched chicks with < 10 organisms of S. enteritidis LA5 or CVL30 was followed by multiplication in the cecal contents. Within 3 days of hatching, the pH of the cecal contents was reduced from ca. 7 to 5. Samples of gut contents were inoculated in vitro. The S. enteritidis strains multiplied in samples taken from the ileum and duodenum irrespective of age but multiplied in the cecal samples from newly hatched chicks only. Invasion from the gut by S. enteritidis LA5 and CVL30 was both age and dose dependent.

Infectious laryngotracheitis (ILT) was diagnosed as the cause of an outbreak of respiratory disease in broiler chickens in California. The classical form of ILT is characterized by dyspnea, gasping, coughing, and expectoration of bloody exudate. Most of the broilers submitted to the diagnostic laboratory showed a non-classical presentation of ILT, in which mucoid tracheitis and conjunctivitis were the most consistent lesions. Historically, most of the ILT cases diagnosed in our laboratory have consisted of layers with classical signs and lesions. It is not known whether this non classical presentation of ILT in broilers is due to differences in the way broilers respond to ILT infection or to the nature of the ILT virus isolate.

An outbreak of spirochetosis occurred in a flock of 75 game chickens in California during fall 1991. Affected birds were weak and anemic. Many had greenish diarrhea. Spirochetes were seen on Giemsa-stained blood smears and in silver-stained tissue sections of kidney, liver, and spleen. Splenomegaly, which is reported to be characteristic of fowl spirochetosis, was not observed in two acutely infected chickens.