An Aptamer-Based Proteomic Analysis of Plasma from Cats (Felis catus) with Clinical Feline Infectious Peritonitis.

Feline infectious peritonitis (FIP) is a systemic disease manifestation of feline coronavirus (FCoV) and is the most important cause of infectious disease-related deaths in domestic cats. FIP has a variable clinical manifestation but is most often characterized by widespread vasculitis with visceral involvement and/or neurological disease that is typically fatal in the absence of antiviral therapy. Using an aptamer-based proteomics assay, we analyzed the plasma protein profiles of cats who were naturally infected with FIP (n = 19) in comparison to the plasma protein profiles of cats who were clinically healthy and negative for FCoV (n = 17) and cats who were positive for the enteric form of FCoV (n = 9). We identified 442 proteins that were significantly differentiable; in total, 219 increased and 223 decreased in FIP plasma versus clinically healthy cat plasma. Pathway enrichment and associated analyses showed that differentiable proteins were related to immune system processes, including the innate immune response, cytokine signaling, and antigen presentation, as well as apoptosis and vascular integrity. The relevance of these findings is discussed in the context of previous studies. While these results have the potential to inform diagnostic, therapeutic, and preventative investigations, they represent only a first step, and will require further validation.

Anti-tumor activity of a T-helper 1 multiantigen vaccine in a murine model of prostate cancer.

Prostate cancer is one of the few malignancies that includes vaccination as a treatment modality. Elements of an effective cancer vaccine should include the ability to elicit a Type I T-cell response and target multiple antigenic proteins expressed early in the disease. Using existing gene datasets encompassing normal prostate tissue and tumors with Gleason Score ≤ 6 and ≥ 8, 10 genes were identified that were upregulated and conserved in prostate cancer regardless of the aggressiveness of disease. These genes encoded proteins also expressed in prostatic intraepithelial neoplasia. Putative Class II epitopes derived from these proteins were predicted by a combination of algorithms and, using human peripheral blood, epitopes which selectively elicited IFN-γ or IL-10 dominant antigen specific cytokine secretion were determined. Th1 selective epitopes were identified for eight antigens. Epitopes from three antigens elicited Th1 dominant immunity in mice; PSMA, HPN, and AMACR. Each single antigen vaccine demonstrated significant anti-tumor activity inhibiting growth of implanted Myc-Cap cells after immunization as compared to control. Immunization with the combination of antigens, however, was superior to each alone in controlling tumor growth. When vaccination occurred simultaneously to tumor implant, multiantigen immunized mice had significantly smaller tumors than controls (p = 0.002) and a significantly improved overall survival (p = 0.0006). This multiantigen vaccine shows anti-tumor activity in a murine model of prostate cancer.

Type I T cells sensitize treatment refractory tumors to chemotherapy through inhibition of oncogenic signaling pathways.

BACKGROUND: The most common clinical outcome observed after treatment with immune checkpoint inhibitor antibodies is disease stabilization. Using vaccines to generate high levels of tumor antigen-specific T-helper 1 (Th1), we show that tumors not eradicated by vaccination demonstrate prolonged disease stabilization. We evaluated the mechanism by which type I T cells inhibit disease progression and potentially influence the subsequent clinical response to standard therapy in treatment refractory cancers. METHODS: We employed a meta-analysis of studies with tumor growth from four different vaccines in two different mammary cancer models. The T-cell subtype and cytokine essential for vaccine-induced tumor inhibition was determined by in vivo neutralization studies and immunohistochemistry. The role of interferon gamma (IFN-γ) in receptor tyrosine kinase and downstream signaling was determined by immunoblotting. The role of suppressor of cytokine signaling 1 (SOCS1) on IFN-γ signaling was evaluated on SOCS1-silenced cells with immunoblotting and immunoprecipitation. The effect of vaccination on growth factor receptor signaling pathways, performed in both luminal (TgMMTVneu) and basal (C3(1)-Tag) mammary cancer models treated with paclitaxel or an anti-HER2-neu monoclonal antibody were assessed via immunoblotting. RESULTS: Immunization with an epitope-based vaccine targeting a representative tumor antigen resulted in elevated tumor trafficking Tbet+CD4 T cells, decreased tumor proliferation and increased apoptosis compared with control vaccinated mice. The resulting disease stabilization was dependent on IFN-γ-secreting CD4+ T cells. In the presence of excess IFN-γ, SOCS1 became upregulated in tumor cells, bound insulin receptor, insulin like growth factor receptor 1 and epidermal growth factor receptor resulting in profound oncogenic signaling inhibition. Silencing SOCS1 restored growth factor receptor signaling and proliferation and prevented cell death. Similar signaling perturbations were detected in vaccinated mice developing antigen-specific Th1 cells. Vaccination synergized with standard therapies and restored disease sensitivity to treatment with both a neu-specific antibody and paclitaxel in TgMMTVneu and to paclitaxel in C3(1)-Tag. Combination of vaccination and chemotherapy or biological therapy was more effective than monotherapy alone in either model and resulted in complete resolution of disease in some individuals. CONCLUSIONS: These data suggest the clinical activity of type I T cells extends beyond direct tumor killing and immune therapies designed to increase type I T cells and could be integrated into standard chemotherapy regimens to enhance therapeutic efficacy.

Disseminated Neospora caninum infection in a dog with severe colitis.

A 12-y-old spayed female Schipperke dog with a previous diagnosis of inflammatory bowel disease was presented with a 2-mo history of severe colitis. The patient's condition progressed to hepatopathy, pneumonia, and dermatitis following management with prednisolone and dexamethasone sodium phosphate. Colonic biopsies identified severe necrosuppurative colitis with free and intracellular parasitic zoites. Postmortem examination confirmed extensive chronic-active ulcerative colitis, severe acute necrotizing hepatitis and splenitis, interstitial pneumonia, ulcerative dermatitis, myelitis (bone marrow), and mild meningoencephalitis with variable numbers of intracellular and extracellular protozoal zoites. PCR on samples of fresh colon was positive for Neospora caninum. Immunohistochemistry identified N. caninum tachyzoites in sections of colon, and a single tissue cyst in sections of brain. Administration of immunosuppressive drugs may have allowed systemic dissemination of Neospora from the intestinal tract.

Ruptured hepatic artery aneurysm in a domestic yak.

A 19.5-y-old, male domestic yak ( Bos grunniens) with a history of sudden unexpected death was submitted for autopsy. The yak had hemoabdomen, and a large blood clot was attached to the liver and forestomachs. The hepatic artery had a saccular aneurysm with a 1-cm tear. The arterial wall at the site of the rupture was thin, and the luminal surface was roughened with yellow streaks. The arterial wall adjacent to the rupture was thickened, white, firm, and less elastic than normal arterial walls. Cause of death was concluded to be acute exsanguination. Similar cases with sudden death have been reported in domestic cattle and humans. No risk factors, such as nutritional deficiencies, genetic predisposition, or blunt trauma, were identified in this case, and there was no gross or histologic evidence of generalized vascular disease.

Strengthening the American Academy of Pediatric Dentistry's Public Policy Advocate Network: Identifying Advocacy Efforts and Recommendations.

PURPOSE: The purposes of this study were to collect information on involvement, training, and barriers to participation in advocacy efforts for Public Policy Advocates (PPAs) of the American Academy of Pediatric Dentistry (AAPD) and make recommendations to the AAPD. METHODS: Preliminary data were collected from the PPAs during structured AAPD program meetings, conference calls, and individual interviews. Based on these data, a survey was created, piloted, and sent electronically to all PPAs. Data were analyzed and collated by frequencies. RESULTS: Responses from 38 PPAs (100 percent) revealed they were involved with state legislatures and state chapters of the AAPD and American Dental Association. Eighty-two percent of the PPAs requested additional public policy training and clearer communication channels within the network. PPAs are funding their own advocacy efforts, and the time and resources spent away from patient care is a financial barrier. CONCLUSIONS: The Public Policy Advocate network holds a broad policy skill set and voluntarily commits time and resource to advocate for the support of the pediatric dental patient at state and federal government levels. The American Academy of Pediatric Dentistry can strengthen the PPA's self-directed leadership role at state and federal levels through formalized training, restructuring of the network, and increased resources.

Embolic mycotic encephalitis in a cow following Mortierella wolfii infection of a surgery site.

A 5-y-old Holstein dairy cow had surgery for a suspected displaced abomasum 10 d postpartum, developed acute neurologic signs at day 19, and was found dead 21 d postpartum. At autopsy, there was a peri-incisional intramuscular abscess that communicated with the peritoneal cavity, as well as hemorrhage and malacia involving the brain, and multiple nodules in the liver, kidneys, and lungs. Fungal hyphae were seen histologically at the surgery site, on the surface of the liver, and in lesions of severe necrotizing vasculitis in the lungs, kidneys, brain, and liver. The uterus was free of fungal organisms. Pan-fungal PCR and DNA sequencing identified the fungus as Mortierella wolfii. Previously reported deaths from M. wolfii have been related to abortion, but in this case, there was no histologic evidence of fungal organisms in the uterus, calving was routine, and there was a several week delay between calving and development of neurologic signs. The findings suggested a unique case of surgical site infection with subsequent embolic mycosis.

Profound Intellectual Disability and the Bestowment View of Moral Status.

This article engages with debates concerning the moral worth of human beings with profound intellectual and multiple disabilities (PIMDs). Some argue that those with such disabilities are morally less valuable than so-called normal human beings, whereas others argue that all human beings have equal moral value and that, therefore, each group of humans ought to be treated with equal concern. We will argue in favor of a view that takes points from opposing camps in the debates about the moral worth of humans with such disabilities. Our position, roughly, is this: most humans with PIMDs are persons in the morally significant sense and, therefore, deserve moral consideration equal to that granted to so-called "normal" human beings. Some humans with PIMD may not be persons, but nevertheless deserve moral consideration equal to that of persons because they stand in a special relation to persons.

Immunization against HIF-1α Inhibits the Growth of Basal Mammary Tumors and Targets Mammary Stem Cells In Vivo.

Purpose: Triple-negative breast cancer (TNBC) represents a cancer stem cell-enriched phenotype. Hypoxia-inducible factor-1α (HIF-1α) induces the expression of proteins associated with stemness and is highly upregulated in TNBC. We questioned whether HIF-1α was immunogenic and whether vaccination targeting HIF-1α would impact the growth of basal-like mammary tumors in transgenic mice.Experimental Design: We evaluated HIF-1α-specific IgG in sera from controls and patients with breast cancer. Class II epitopes derived from the HIF-1α protein sequence were validated by ELISPOT. To assess therapeutic efficacy, we immunized Tg-MMTVneu and C3(1)Tag mice with HIF-1α Th1-inducing peptides. Stem cells were isolated via magnetic bead separation. Levels of HIF-1α and stem cells in the tumor were quantitated by Western blotting and flow cytometry.Results: The magnitude (P < 0.001) and incidence (P < 0.001) of HIF-1α-specific IgG were elevated in TNBC patients compared with controls. Both breast cancer patients and donors showed evidence of HIF-1α-specific Th1 and Th2 immunity. Three HIF-1α-specific Th1 class II restricted epitopes that were highly homologous between species elicited type I immunity in mice. After HIF-1α vaccination, mammary tumor growth was significantly inhibited in only C3(1)Tag (basal-like/stem cellhigh; P < 0.001) not TgMMTV-neu (luminal/neu/stem celllow; P = 0.859) murine models. Vaccination increased type I T cells in the tumor (P = 0.001) and decreased cells expressing the stem cell marker, Sca-1, compared with controls (P = 0.004).Conclusions: An HIF-1α vaccine may be uniquely effective in limiting tumor growth in TNBC. Inhibiting outgrowth of breast cancer stem cells via active immunization in the adjuvant setting may impact disease recurrence. Clin Cancer Res; 23(13); 3396-404. ©2016 AACR.

The Moorean argument for the full moral status of those with profound intellectual disability: a rejoinder to Roberts.

In a recent paper we argued that a Moorean strategy can be employed to justify our continuing to believe the following proposition, even in the presence of philosophical views that entail it is false, without any philosophical argument against those views, and without any positive philosophical argument in its favour: H>A: Humans have an equal moral status that is higher than the moral status of non-human animals. The basic idea is that our confidence in the truth of this proposition is greater than our confidence in the propositions that make up those philosophical views that entail that it is false, and that this is sufficient to justify rejecting those views and to continue to believe H>A. Roberts has recently responded to our argument by claiming that (i) although the Moorean strategy is valid, it is not powerful and (ii) a resort to the Moorean strategy reflects too great a pessimism about the accounts available that purport to justify H>A. In this short rejoinder we explain why Roberts fails to establish his two claims.

A Moorean argument for the full moral status of those with profound intellectual disability.

This paper is about the moral status of those human beings who have profound intellectual disabilities (PIDs). We hold the common sense view that they have equal status to 'normal' human beings, and a higher status than any non-human animal. On the standard account of moral status, this view cannot be sustained. In this paper, we ask whether, in order to be justified in continuing to hold our view, we are obliged to offer an alternative account that does sustain it? Our answer is that we are not.

Fixation of Metacarpal Shaft Fractures: Biomechanical Comparison of Intramedullary Nail Crossed K-Wires and Plate-Screw Constructs.

OBJECTIVES: Metacarpal (MC) fractures are very common, accounting for 18% of all fractures distal to the elbow. Many MC fractures can be treated non-operatively; however, some are treated most effectively with surgical stabilization, for which there are multiple methods. It was postulated that plates would have a significantly higher (P < 0.05) load to failure than crossed K(XK)-wires and that intramedullary metacarpal nails (IMNs) and XK-wires would have equivalent load to failure. METHODS: Mid-diaphyseal transverse fractures were created in 36 synthetic metacarpals and stabilized using nails, XK-wires or non-locking plates. Three-point bending was performed with continuous recording of load and displacement. Statistical analysis was performed using single factor ANOVA and Scheffe's test. Statistical significance was defined as P < 0.05. RESULTS: Biomechanical testing revealed significant differences between groups in load-to-failure. Average load to failure was significantly greater in the plate (1669 ± 322 N) than the XK-wire (146 ± 56 N) or IMN (110 ± 43 N) groups. The loads to failure of the K-wires and nails were equivalent. Plates were 11 and 15 times stronger in three-point bending than the K-wires and nails, respectively. There was no statistically significant difference between strengths of the K-wires and nails. CONCLUSIONS: Although plates are the most stable means of fixation of midshaft metacarpal fractures, if minimally-invasive techniques are indicated, intramedullary nails may provide equivalent stability as commonly-used XK-wires. Although some studies have shown favorable clinical outcomes with IMNs, additional clinical correlation of these biomechanical results to fracture healing and outcomes is needed.

The Krüppel-like factor 2 and Krüppel-like factor 4 genes interact to maintain endothelial integrity in mouse embryonic vasculogenesis.

BACKGROUND: Krüppel-like Factor 2 (KLF2) plays an important role in vessel maturation during embryonic development. In adult mice, KLF2 regulates expression of the tight junction protein occludin, which may allow KLF2 to maintain vascular integrity. Adult tamoxifen-inducible Krüppel-like Factor 4 (KLF4) knockout mice have thickened arterial intima following vascular injury. The role of KLF4, and the possible overlapping functions of KLF2 and KLF4, in the developing vasculature are not well-studied. RESULTS: Endothelial breaks are observed in a major vessel, the primary head vein (PHV), in KLF2-/-KLF4-/- embryos at E9.5. KLF2-/-KLF4-/- embryos die by E10.5, which is earlier than either single knockout. Gross hemorrhaging of multiple vessels may be the cause of death. E9.5 KLF2-/-KLF4+/- embryos do not exhibit gross hemorrhaging, but cross-sections display disruptions of the endothelial cell layer of the PHV, and these embryos generally also die by E10.5. Electron micrographs confirm that there are gaps in the PHV endothelial layer in E9.5 KLF2-/-KLF4-/- embryos, and show that the endothelial cells are abnormally bulbous compared to KLF2-/- and wild-type (WT). The amount of endothelial Nitric Oxide Synthase (eNOS) mRNA, which encodes an endothelial regulator, is reduced by 10-fold in E9.5 KLF2-/-KLF4-/- compared to KLF2-/- and WT embryos. VEGFR2, an eNOS inducer, and occludin, a tight junction protein, gene expression are also reduced in E9.5 KLF2-/-KLF4-/- compared to KLF2-/- and WT embryos. CONCLUSIONS: This study begins to define the roles of KLF2 and KLF4 in the embryonic development of blood vessels. It indicates that the two genes interact to maintain an intact endothelial layer. KLF2 and KLF4 positively regulate the eNOS, VEGFR2 and occludin genes. Down-regulation of these genes in KLF2-/-KLF4-/- embryos may result in the observed loss of vascular integrity.

A zygote could be a human: a defence of conceptionism against fission arguments.

In this paper I defend the view that a zygote is a human from the fission objection that is widely thought to be decisive against the view. I do so, drawing upon a recent discussion of this issue by John Burgess, by explaining in detail the metaphysical position the proponent of the view should adopt in order to rebut the objection.

Characterization of the anchoring morphology and mineral content of the anterior cruciate and medial collateral ligaments of the knee.

The manner in which ligament connects to bone remains an area of interest for researchers, bioengineers, and clinicians. Stable fixation of an anterior cruciate ligament (ACL) graft has been shown to be paramount to preventing excess anterior tibial translation and to restoring the normal kinematics of the knee joint. In this study, the surface area of attachment and the mineral characteristics of the ACL and medial collateral ligament (MCL) attachment sites were characterized to determine the factors that contributed to ligament attachment strength. Findings from this study indicated that the area of attachment of the ACL's insertion was significantly greater than the ligament's origin (95.8 mm(2) ± 21.5 vs. 73.2 mm(2) ± 16.2, P = 0.009). Additionally, the ACL was measured to have a greater surface area of attachment when compared with the MCL (84.5 mm(2) ± 18.8 vs. 58.2 mm(2) ± 23.8, P = 0.005); although, the MCL was observed to have a greater region of calcified fibrocartilage (CFC) than the ACL (533.0 µm ± 116.9 vs. 195.5 µm ± 36.6, P = 0.0003). No significant correlation was observed between the ligament's area of attachment and the thickness of the CFC region. Measurements of ash percent suggested that the boundary region, between the CFC and host bone, possessed the least mineral content for the three regions of interest. These data suggest that ligament attachment strength can be attributed to several factors, including the ligament's area of attachment, regional thickness, and mineral content of the CFC.

The effect of sports injury on insulin-like growth factor-I and type 3 procollagen: implications for detection of growth hormone abuse in athletes.

CONTEXT: A method to detect exogenously administered growth hormone (GH) based on the measurement of two GH-dependent markers, IGF-I and type 3 procollagen (P-III-P) has been proposed. Skeletal or soft tissue injury may alter these markers. Elevations in either of these proteins after injury might lead to a false accusation of doping with GH. OBJECTIVE: The objective of the study was to assess the effect of musculoskeletal or soft tissue injury on IGF-I and P-III-P concentrations in amateur and elite athletes and assess the effect of injury on the proposed GH detection method. DESIGN: This was a longitudinal observational study after sporting injury. SETTING: The study was conducted at Southampton General Hospital and British Olympic Medical Centre. SUBJECTS: Subjects included elite and amateur athletes after an injury. INTERVENTION: Interventions included measurement of IGF-I and P-III-P and application of the GH-2000 discriminant function score up to 84 d after an injury as well as classification of injury by type and severity. OUTCOME MEASURES: IGF-I and P-III-P concentration and ability to detect GH abuse in athletes without the risk of false accusation because of an injury were measured. RESULTS: There was no change in IGF-I concentration after an injury. By contrast, P-III-P concentrations rose by 41.1 +/- 16.6%, reaching a peak around 14 d after an injury. The rise in P-III-P varied according to injury type and severity. This rise had a trivial effect on the GH-2000 discriminant function score, and no subject reached the threshold needed for a doping offense. CONCLUSIONS: Although there was a rise in P-III-P after injury, this was insufficient to invalidate the GH-2000 detection method based on IGF-I and P-III-P concentrations.