Calf health and performance during receiving is not changed by fence-line preconditioning on flint hills range vs. drylot preconditioning

Ranch-of-origin preconditioning can improve the welfare and performance of beef calves by decreasing the stress associated with weaning, transport, diet change, and commingling with other calves. Preconditioning methods that involve pasture weaning coupled with maternal contact (i.e., fence-line weaning) have been promoted as possible best management practices for minimizing stress. Prior studies focused on performance and behavior during preconditioning on the ranch of origin. Little information has been published relating to carryover effects of fence-line preconditioning compared with conventional drylot preconditioning on performance and behavior during feedlot receiving. Our objectives were to measure growth and health during a 28-day ranch-of-origin preconditioning phase and during a 60-day feedlot receiving phase among beef calves subjected to 1 of 3 ranch-of-origin preconditioning programs: (1) drylot preconditioning + dam separation, (2) pasture preconditioning + fence-line contact with dams, and (3) pasture preconditioning + fence-line contact with dams + supplemental feed delivered in a bunk. In addition, we recorded incidences of behavioral distress among these treatments during first 7 days of feedlot receiving

Defining Breadth of Hepatitis C Virus Neutralization

Extraordinary genetic diversity is a hallmark of hepatitis C virus (HCV). Therefore, accurate measurement of the breadth of antibody neutralizing activity across diverse HCV isolates is key to defining correlates of immune protection against the virus, and essential to guide vaccine development. Panels of HCV pseudoparticle (HCVpp) or replication-competent cell culture viruses (HCVcc) can be used to measure neutralizing breadth of antibodies. These in vitro assays have been used to define neutralizing breadth of antibodies in serum, to characterize broadly neutralizing monoclonal antibodies, and to identify mechanisms of HCV resistance to antibody neutralization. Recently, larger and more diverse panels of both HCVpp and HCVcc have been described that better represent the diversity of circulating HCV strains, but further work is needed to expand and standardize these neutralization panels

Maintenance of viral suppression in HIV-1–infected HLA-B*57+ elite suppressors despite CTL escape mutations

Rare human immunodeficiency virus 1–infected individuals, termed elite suppressors (ES), maintain plasma virus levels of <50 copies/ml and normal CD4 counts without therapy. The major histocompatibility complex class I allele group human histocompatibility leukocyte antigen (HLA)-B*57 is overrepresented in this population. Mutations in HLA-B*57–restricted epitopes have been observed in ES, but their significance has remained unclear. Here we investigate the extent and impact of cytotoxic T lymphocyte (CTL) escape mutations in HLA-B*57+ ES. We provide the first direct evidence that most ES experience chronic low level viremia. Sequencing revealed a striking discordance between the genotypes of plasma virus and archived provirus in resting CD4+ T cells. Mutations in HLA-B*57–restricted Gag epitopes were present in all viruses from plasma but were rare in proviruses, suggesting powerful selective pressure acting at these epitopes. Surprisingly, strong CD8+ T cell interferon-γ responses were detected against some mutant epitopes found in plasma virus, suggesting the development of de novo responses to viral variants. In some individuals, relative CD8+ T cell interleukin-2 responses showed better correlation with the selection observed in vivo. Thus, analysis of low level viremia reveals an unexpectedly high level of CTL escape mutations reflecting selective pressure acting at HLA-B*57–restricted epitopes in ES. Continued viral suppression probably reflects CTL responses against unmutated epitopes and residual or de novo responses against epitopes with escape mutations

Self-Adjuvanting Glycopeptide Conjugate Vaccine against Disseminated Candidiasis

Our research on pathogenesis of disseminated candidiasis led to the discovery that antibodies specific for Candida albicans cell surface β-1, 2–mannotriose [β-(Man)3] protect mice. A 14 mer peptide Fba, which derived from the N-terminal portion of the C. albicans cytosolic/cell surface protein fructose-bisphosphate aldolase, was used as the glycan carrier and resulted in a novel synthetic glycopeptide vaccine β-(Man)3-Fba. By a dendritic cell-based immunization approach, this conjugate induced protective antibody responses against both the glycan and peptide parts of the vaccine. In this report, we modified the β-(Man)3-Fba conjugate by coupling it to tetanus toxoid (TT) in order to improve immunogenicity and allow for use of an adjuvant suitable for human use. By new immunization procedures entirely compatible with human use, the modified β-(Man)3-Fba-TT was administered either alone or as a mixture made with alum or monophosphoryl lipid A (MPL) adjuvants and given to mice by a subcutaneous (s.c.) route. Mice vaccinated with or, surprisingly, without adjuvant responded well by making robust antibody responses. The immunized groups showed a high degree of protection against a lethal challenge with C. albicans as evidenced by increased survival times and reduced kidney fungal burden as compared to control groups that received only adjuvant or DPBS buffer prior to challenge. To confirm that induced antibodies were protective, sera from mice immunized against the β-(Man)3-Fba-TT conjugate transferred protection against disseminated candidiasis to naïve mice, whereas C. albicans-absorbed immune sera did not. Similar antibody responses and protection induced by the β-(Man)3-Fba-TT vaccine was observed in inbred BALB/c and outbred Swiss Webster mice. We conclude that addition of TT to the glycopeptide conjugate results in a self-adjuvanting vaccine that promotes robust antibody responses without the need for additional adjuvant, which is novel and represents a major step forward in vaccine design against disseminated candidiasis

Mid-Infrared High-Contrast Imaging of HD 114174 B : An Apparent Age Discrepancy in a "Sirius-Like" Binary System

We present new observations of the faint "Sirius-like" companion discovered to orbit HD 114174. Previous attempts to image HD 114174 B at mid-infrared wavelengths using NIRC2 at Keck have resulted in a non-detection. Our new L'-band observations taken with the Large Binocular Telescope and LMIRCam recover the companion ($\Delta L$ = 10.15 $\pm$ 0.15 mag, $\rho$ = 0.675'' $\pm$ 0.016'') with a high signal-to-noise ratio (10 $\sigma$). This measurement represents the deepest L' high-contrast imaging detection at sub-arcsecond separations to date, including extrasolar planets. We confirm that HD 114174 B has near-infrared colors consistent with the interpretation of a cool white dwarf ($J-L'$ = 0.76 $\pm$ 0.19 mag, $K-L'$ = 0.64 $\pm$ 0.20). New model fits to the object's spectral energy distribution indicate a temperature $T_{\rm eff}$ = 4260 $\pm$ 360 K, surface gravity log g = 7.94 $\pm$ 0.03, a cooling age t$_{c} \approx$ 7.8 Gyr, and mass $M$ = 0.54 $\pm$ 0.01 $M_{\odot}$. We find that the cooling age given by theoretical atmospheric models do not agree with the age of HD 114174 A derived from both isochronological and gyrochronological analyses. We speculate on possible scenarios to explain the apparent age discrepancy between the primary and secondary. HD 114174 B is a nearby benchmark white dwarf that will ultimately enable a dynamical mass estimate through continued Doppler and astrometric monitoring. Efforts to characterize its physical properties in detail will test theoretical atmospheric models and improve our understanding of white dwarf evolution, cooling, and progenitor masses.Comment: 6 pages, 3 figures, to be published in the Astrophysical Journal Letter

Nuclear Retention of Multiply Spliced HIV-1 RNA in Resting CD4(+) T Cells

HIV-1 latency in resting CD4(+) T cells represents a major barrier to virus eradication in patients on highly active antiretroviral therapy (HAART). We describe here a novel post-transcriptional block in HIV-1 gene expression in resting CD4(+) T cells from patients on HAART. This block involves the aberrant localization of multiply spliced (MS) HIV-1 RNAs encoding the critical positive regulators Tat and Rev. Although these RNAs had no previously described export defect, we show that they exhibit strict nuclear localization in resting CD4(+) T cells from patients on HAART. Overexpression of the transcriptional activator Tat from non-HIV vectors allowed virus production in these cells. Thus, the nuclear retention of MS HIV-1 RNA interrupts a positive feedback loop and contributes to the non-productive nature of infection of resting CD4(+) T cells. To define the mechanism of nuclear retention, proteomic analysis was used to identify proteins that bind MS HIV-1 RNA. Polypyrimidine tract binding protein (PTB) was identified as an HIV-1 RNA-binding protein differentially expressed in resting and activated CD4(+) T cells. Overexpression of PTB in resting CD4(+) T cells from patients on HAART allowed cytoplasmic accumulation of HIV-1 RNAs. PTB overexpression also induced virus production by resting CD4(+) T cells. Virus culture experiments showed that overexpression of PTB in resting CD4(+) T cells from patients on HAART allowed release of replication-competent virus, while preserving a resting cellular phenotype. Whether through effects on RNA export or another mechanism, the ability of PTB to reverse latency without inducing cellular activation is a result with therapeutic implications

HCV Broadly Neutralizing Antibodies Use a CDRH3 Disulfide Motif to Recognize an E2 Glycoprotein Site that Can Be Targeted for Vaccine Design

Hepatitis C virus (HCV) vaccine efforts are hampered by the extensive genetic diversity of HCV envelope glycoproteins E1 and E2. Structures of broadly neutralizing antibodies (bNAbs) (e.g., HEPC3, HEPC74) isolated from individuals who spontaneously cleared HCV infection facilitate immunogen design to elicit antibodies against multiple HCV variants. However, challenges in expressing HCV glycoproteins previously limited bNAb-HCV structures to complexes with truncated E2 cores. Here we describe crystal structures of full-length E2 ectodomain complexes with HEPC3 and HEPC74, revealing lock-and-key antibody-antigen interactions, E2 regions (including a target of immunogen design) that were truncated or disordered in E2 cores, and an antibody CDRH3 disulfide motif that exhibits common interactions with a conserved epitope despite different bNAb-E2 binding orientations. The structures display unusual features relevant to common genetic signatures of HCV bNAbs and demonstrate extraordinary plasticity in antibody-antigen interactions. In addition, E2 variants that bind HEPC3/HEPC74-like germline precursors may represent candidate vaccine immunogens