Germline mutation landscape of DNA damage repair genes in African Americans with prostate cancer highlights potentially targetable RAD genes.

In prostate cancer, emerging data highlight the role of DNA damage repair genes (DDRGs) in aggressive forms of the disease. However, DDRG mutations in African American men are not yet fully defined. Here, we profile germline mutations in all known DDRGs (N = 276) using whole genome sequences from blood DNA of a matched cohort of patients with primary prostate cancer comprising of 300 African American and 300 European Ancestry prostate cancer patients, to determine whether the mutation status can enhance patient stratification for specific targeted therapies. Here, we show that only 13 of the 46 DDRGs identified with pathogenic/likely pathogenic mutations are present in both African American and European ancestry patients. Importantly, RAD family genes (RAD51, RAD54L, RAD54B), which are potentially targetable, as well as PMS2 and BRCA1, are among the most frequently mutated DDRGs in African American, but not in European Ancestry patients.

Rotavirus A Associated with Clinical Disease and Hepatic Necrosis in California Pigeons (Columba livia domestica).

Retrospective analysis of pigeon necropsy submissions to the California Animal Health and Food Safety Laboratory System from 2000 to 2018 revealed 14 submissions diagnosed with rotavirus A hepatic necrosis or "reoviruslike" viral hepatitis. Nine of the 14 submissions (64%) occurred in 2018. Submissions were racing pigeons and squab breeders from flocks with increased mortality. Juvenile and adult pigeons were submitted with a history of depression, diarrhea, regurgitation, labored breathing, and weakness. Flock morbidity peaked at 80% and mortality at 28%. The most consistent findings on postmortem examination were variably congested, mottled, and enlarged livers and spleens. Microscopically, mild to severe hepatic necrosis was observed with variable bile duct hyperplasia, sinusoidal congestion, hemosiderosis, and portal lymphoplasmacytic inflammation. Rotavirus A was detected in hepatocytes and inflammatory cells by immunohistochemistry. Negative-stain electron microscopy identified viral particles consistent with a member of Reoviridae in all negatively stained liver homogenates. Eleven cases were analyzed by reverse transcriptase-PCR targeting rotavirus A viral protein (VP) 6 and VP7 genes. Subsequent phylogenetic analysis of the VP6 and VP7 sequences compared to published Chinese, Nigerian, and German rotavirus A VP6 and VP7 sequences demonstrated the formation of two and three distinct clades, respectively. To the authors' knowledge, rotavirus A hepatic necrosis in pigeons has not been previously reported in the United States and represents a significant emerging disease for the pigeon industry due to the potential for high flock mortality and lost production.

Rotavirus A asociado con enfermedad clínica y necrosis hepática en palomas de California (Columba livia domestica). El análisis retrospectivo de los casos de necropsias de palomas remitidos al Sistema de Laboratorio de Salud Animal y Seguridad Alimentaria del Estado de California entre los años 2000 a 2018 reveló 14 casos con diagnóstico de necrosis hepática por rotavirus A, o hepatitis viral ocasionada por "virus similares a reovirus". Nueve de los 14 casos (64%) ocurrieron en el año 2018. Los casos fueron de palomas de competencia y de criadores de pichones de parvadas con aumento en la mortalidad. Se presentaron palomas jóvenes y adultas con antecedentes de depresión, diarrea, regurgitación, dificultad para respirar y debilidad. La morbilidad mayor fue de un 80% como máximo y la mortalidad fue de un 28%. Los hallazgos más consistentes en el examen post mortem incluyeron hígados y bazos con congestión, apariencia moteada y aumento de tamaño de forma variable. Microscópicamente, se observó necrosis hepática de leve a severa con hiperplasia variable de los conductos biliares, congestión de sinusoides, hemosiderosis e inflamación linfoplasmocítica portal. Se detectó rotavirus A en hepatocitos y células inflamatorias por inmunohistoquímica. La microscopía electrónica de tinción negativa identificó partículas virales consistentes con virus posiblemente miembros de la familia Reoviridae en todos los homogenizados de hígado teñidos negativamente. Se analizaron once casos mediante transcripción reversa y PCR dirigida a los genes de la proteína viral (VP) 6 y VP7 del rotavirus A. El análisis filogenético posterior de las secuencias de los genes VP6 y VP7 cuando se compararon con secuencias de genes VP6 y VP7 de rotavirus A de China, Nigeria y de Alemania previamente publicadas demostró la formación de dos y tres clados distintos, respectivamente. De acuerdo con el conocimiento de los autores, la necrosis hepática por rotavirus A en palomas no se había reportado previamente en los Estados Unidos y representa una enfermedad emergente importante para la industria de las palomas debido a su potencial de alta mortalidad de la parvada y a las pérdidas en la producción.

The Leukocyte Esterase Test Strip Is a Poor Rule-Out Test for Periprosthetic Joint Infection.

BACKGROUND: The urinary leukocyte esterase (LE) test strip has been suggested as a good screening test for periprosthetic joint infection (PJI). The purpose of this study is to compare the diagnostic profile of LE assays from different manufacturers and determine whether the LE test strip is a good rule-out test. METHODS: Synovial fluid samples (N = 344), sent to 1 laboratory for PJI testing, were used in this prospective study. Four different tests for synovial fluid LE were simultaneously evaluated for their performance in detecting white blood cell (WBC) positive samples (>3000 cells/µL). RESULTS: Both neutrophil elastase immunoassays demonstrated greater sensitivity than urinary LE test strips (92.0% and 90.8% vs 72.4% and 80.3%; all P < 0.011). Fifty-three percent of false-negative urinary LE test strip results clearly missed the presence of elevated levels of synovial fluid LE. Invalid urinary LE test strip results were 4-fold more likely among WBC (+) compared with WBC (-) samples (27.0% vs 6.8%; P < 0.0001). The combined failure to detect an elevated WBC count, because of either false-negative or invalid results, was 47.1% and 41.4% for the Roche and Siemens test strips, respectively. CONCLUSIONS: This study agrees with the existing literature demonstrating that the LE test strips are among the lowest sensitivity tests for PJI. The urinary LE tests strips should not be used to rule-out PJI, as they often fail to detect abundant levels of LE in synovial fluid. Instead, it is more appropriate to use the (++) LE test strip result as a secondary confirmatory rule-in test for PJI because of its high specificity.

Application of high-intensity focused ultrasound to the study of mild traumatic brain injury.

Though intrinsically of much higher frequency than open-field blast overpressures, high-intensity focused ultrasound (HIFU) pulse trains can be frequency modulated to produce a radiation pressure having a similar form. In this study, 1.5-MHz HIFU pulse trains of 1-ms duration were applied to intact skulls of mice in vivo and resulted in blood-brain barrier disruption and immune responses (astrocyte reactivity and microglial activation). Analyses of variance indicated that 24 h after HIFU exposure, staining density for glial fibrillary acidic protein was elevated in the parietal and temporal regions of the cerebral cortex, corpus callosum and hippocampus, and staining density for the microglial marker, ionized calcium binding adaptor molecule, was elevated 2 and 24 h after exposure in the corpus callosum and hippocampus (all statistical test results, p < 0.05). HIFU shows promise for the study of some bio-effect aspects of blast-related, non-impact mild traumatic brain injuries in animals.

Short and long-term motor and behavioral effects of diazoxide and dimethyl sulfoxide administration in the mouse after traumatic brain injury.

Traumatic brain injury (TBI) is a worldwide phenomenon that affects all ages and socioeconomic classes and results in varying degrees of immediate and delayed motor, cognitive, and emotional deficiencies. A plethora of pharmacologic interventions that target recognized initiators and propagators of pathology are being investigated in an attempt to ameliorate secondary injury processes that follow primary injury. Diazoxide (DZ), a K(ATP) channel activator, has been shown to provide short- and long-term protective effects in a variety of in vitro and in vivo cerebral ischemia models. However, the effects of DZ on behavioral outcome following TBI have not been investigated. TBI was induced in male C57BL/6J mice by controlled cortical impact (CCI) and followed by intraperitoneal administration of either normal saline, dimethyl sulfoxide (DMSO), or 2.5 mg/kg DZ in DMSO, 30 min post-injury and daily for three days. Open field and beam walk performances were used to assess motor and behavioral function 1, 7, and 14 days following injury. Spatial learning and memory were assessed three weeks following injury using the Morris water maze. Injured mice were significantly impaired on the beam-walk and Morris water maze tasks, and were hyperactive and anxious in an open field environment. On post-injury days 1 and 14, mice treated with DMSO exhibited an increase in the amount of time required to perform the beam walk task. In addition, animals exposed to DMSO or DZ+DMSO exhibited slower swimming speed in the Morris water maze on the final day of testing. There was no therapeutic effect, however, of the treatment or vehicle on open field behavior or learning and memory function in the Morris water maze. In summary, CCI produced significant long-term impairment of motor, memory, and behavioral performance measures, and DZ administration, under the conditions used, provided no functional benefits following injury.

Mouse brain PSA-NCAM levels are altered by graded-controlled cortical impact injury.

Traumatic brain injury (TBI) is a worldwide endemic that results in unacceptably high morbidity and mortality. Secondary injury processes following primary injury are composed of intricate interactions between assorted molecules that ultimately dictate the degree of longer-term neurological deficits. One comparatively unexplored molecule that may contribute to exacerbation of injury or enhancement of recovery is the posttranslationally modified polysialic acid form of neural cell adhesion molecule, PSA-NCAM. This molecule is a critical modulator of central nervous system plasticity and reorganization after injury. In this study, we used controlled cortical impact (CCI) to produce moderate or severe TBI in the mouse. Immunoblotting and immunohistochemical analysis were used to track the early (2, 24, and 48 hour) and late (1 and 3 week) time course and location of changes in the levels of PSA-NCAM after TBI. Variable and heterogeneous short- and long-term increases or decreases in expression were found. In general, alterations in PSA-NCAM levels were seen in the cerebral cortex immediately after injury, and these reductions persisted in brain regions distal to the primary injury site, especially after severe injury. This information provides a starting point to dissect the role of PSA-NCAM in TBI-related pathology and recovery.

Expression of HLA class II molecules in humanized NOD.Rag1KO.IL2RgcKO mice is critical for development and function of human T and B cells.

BACKGROUND: Humanized mice able to reconstitute a surrogate human immune system (HIS) can be used for studies on human immunology and may provide a predictive preclinical model for human vaccines prior to clinical trials. However, current humanized mouse models show sub-optimal human T cell reconstitution and limited ability to support immunoglobulin class switching by human B cells. This limitation has been attributed to the lack of expression of Human Leukocyte Antigens (HLA) molecules in mouse lymphoid organs. Recently, humanized mice expressing HLA class I molecules have been generated but showed little improvement in human T cell reconstitution and function of T and B cells. METHODS: We have generated NOD.Rag1KO.IL2RγcKO mice expressing HLA class II (HLA-DR4) molecules under the I-E(d) promoter that were infused as adults with HLA-DR-matched human hematopoietic stem cells (HSC). Littermates lacking expression of HLA-DR4 molecules were used as control. RESULTS: HSC-infused HLA-DR4.NOD.Rag1KO.IL-2RγcKO mice developed a very high reconstitution rate (>90%) with long-lived and functional human T and B cells. Unlike previous humanized mouse models reported in the literature and our control mice, the HLA-DR4 expressing mice reconstituted serum levels (natural antibodies) of human IgM, IgG (all four subclasses), IgA, and IgE comparable to humans, and elicited high titers of specific human IgG antibodies upon tetanus toxoid vaccination. CONCLUSIONS: Our study demonstrates the critical role of HLA class II molecules for development of functional human T cells able to support immunoglobulin class switching and efficiently respond to vaccination.

Differential protection against MPTP or methamphetamine toxicity in dopamine neurons by deletion of ppN/OFQ expression.

Nociceptin (N/OFQ) is an endogenous neuropeptide that plays a role in the behavioral deficits associated with Parkinson's disease (PD). The purpose of the present study was to characterize the protective effects of prepro (pp)N/OFQ gene deletion against two dopamine toxins, MPTP and methamphetamine (METH). Results demonstrate that ppN/OFQ gene deletion attenuates the loss of both the number of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra pars compacta (SNpc) and loss of TH and vesicular monoamine transporter-2 (VMAT) immunoreactivity in the caudate putamen (CPu) of MPTP-treated mice. This protection was unaffected by age or gender, although, when loss of TH exceeded 90% in 5-6 month-old mice, the protective effect was greatly diminished. In contrast, METH administration preferentially damaged dopaminergic terminals in the CPu with little effect on dopamine neurons in the SNpc, an effect not reversed by ppN/OFQ gene deletion. To determine if N/OFQ and MPP+ act directly and synergistically on dopamine neurons, differentiated SH-SY5Y cells were incubated with N/OFQ and/or MPP+. N/OFQ did not increase MPP+-mediated cell loss, suggesting an indirect action of N/OFQ. These studies demonstrate that inhibition of the endogenous N/OFQ system may represent a new therapeutic target for prevention of neuronal loss associated with PD.

Envy, shame, and sadism.

In this article, envy is identified as a drive derivative, erupting as a definable behavior in the anal phase of psychosexual development, locating envy as first being experienced during the stage of self-development, separation/individuation. The aim of envy, as identified by the nature of the investment the subject makes in envied objects, is described as "aquisitiveness," or, following Klein, sadism, defined in terms of the subject wanting to incorporate the envied object, making the aim of envy narcissistic, that is, an aggrandizing of the self. The usual aim assigned to envy, destructiveness, is conceptualized as a defense against the cannibalistic hunger of envy, in response to the regulatory affect, shame. The dynamic of this defense accounts for the dyadic nature of the object of envy (as opposed to jealousy), supporting the "splitting" quality of this defense. This rendering of envy, shame, and sadism promotes the integration of narcissistic urges into normative psychosexual development and deepens an understanding of the splitting defense in pathological narcissism.

Distribution of nociceptin/orphanin FQ in adult human brain.

Nociceptin/orphanin FQ (N/OFQ), the endogenous agonist for the opioid receptor-like receptor 1 (ORL1), shows significant similarities to dynorphin A in structure and distribution in rat central nervous system. The distribution of N/OFQ in human brain has not been studied. We measured the concentrations of N/OFQ in 47 microdissected areas of the central nervous system of adult human brain using radioimmunoassay (RIA). Significant heterogeneity was found in the levels of N/OFQ concentration in the various analyzed regions. The highest concentrations were measured in the dorsal central gray matter (periaqueductal gray), the locus coeruleus, the ventromedial nucleus of hypothalamus, the septum and the dorsal horn of the spinal cord. High concentrations were also detected in other hypothamamic nuclei, the inferior colliculus, the ventral central gray matter, the pontine tegmentum, the amygdala, the reticular formation and the spinal trigeminal nucleus. Considerable similarity with the distribution of N/OFQ in rat CNS was observed. The widespread distribution in CNS predicts multifaceted functions for N/OFQ.

Characterization of the antigenic, immunogenic, and pathogenic variation of infectious bursal disease virus due to propagation in different host systems (bursa, embryo, and cell culture). I. Antigenicity and immunogenicity.

In vitro and in ovo virus neutralization assays were conducted to assess the role of different host systems in infectious bursal disease virus (IBDV) antigenic and immunogenic variation. Four different strains, two variant (1084 E and GLS) and two standard (Edgar and STC), were propagated separately in the bursa of Fabricius and embryos, and were compared with cell culture-adapted preparations of the homologous strains. Chicken polyclonal antisera were prepared against each IBDV and neutralizing antibody titres were determined. Normalized IBDV antibody concentrations were used in neutralization assays against homologous and heterologous IBDVs in 10-day-old specific pathogen free embryos. Both antigenic and immunogenic changes occurred in IBDVs evaluated, as evidenced by differences in the ability of normalized antibody to neutralize IBDV propagated in different host systems. Antibody induced by bursal-derived IBDV neutralized all isolates equally well, whereas antibody induced by cell culture-derived virus neutralized bursal-derived IBDV much less effectively.

Characterization of the antigenic, immunogenic, and pathogenic variation of infectious bursal disease virus due to propagation in different host systems (bursa, embryo, and cell culture). II. Antigenicity at the epitope level.

Antigenic variation of infectious bursal disease virus (IBDV) due to propagation in different host systems (bursa of Fabricius, embryos, or cell cultures) was determined by enzyme-linked immunosorbent assay (indirect and antigen capture) and western blot analysis. To conduct this study, we used 27 non-neutralizing anti-VP(2) monoclonal antibodies, a reference panel of nine neutralizing monoclonal antibodies, and 13 neutralizing anti-IBDV chicken polyclonal antibodies. Changes occurred in neutralizing, cross-reactive, conformation-dependent epitopes on the VP(2) protein of IBDV. Interestingly, non-neutralizing, cross-reactive, conformation-dependent and confirmation-independent epitopes also changed on VP(2). These epitope changes were directly associated with the method used to propagate IBDV. These results demonstrate that different host systems may play an important role in the antigenicity of IBDV.

Characterization of the antigenic, immunogenic, and pathogenic variation of infectious bursal disease virus due to propagation in different host systems (bursa, embryo, and cell culture). III. Pathogenicity.

Differences in the relative pathogenicity of variant (1084 E and GLS) and standard (Edgar and STC) infectious bursal disease virus (IBDV) strains were observed after propagation in the bursa of Fabricius, embryos, or cell cultures. Bursa-derived IBDV induced the most severe lesions in the bursa of Fabricius when compared with strains propagated in embryos or cell cultures. Embryo-derived IBDV induced moderate gross bursal lesions, whereas cell culture-derived IBDV did not damage the bursa grossly. A high frequency of virus re-isolations was obtained from bursal, spleen, and thymic samples collected from birds inoculated with bursa-derived or embryo-derived IBDV. Virus re-isolation occurred much less frequently from birds inoculated with cell culture-adapted IBDV. Serological evaluations demonstrated that bursa-derived IBDV strains induced a higher neutralizing antibody response than did embryo-derived or cell culture-derived strains. These results document that the relative pathogenicity and immunogenicity of IBDV is reduced following propagation in embryos or cell cultures.