VLPs generated by the fusion of RSV-F or hMPV-F glycoprotein to HIV-Gag show improved immunogenicity and neutralizing response in mice.

Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) vaccines have been long overdue. Structure-based vaccine design created a new momentum in the last decade, and the first RSV vaccines have finally been approved in older adults and pregnant individuals. These vaccines are based on recombinant stabilized pre-fusion F glycoproteins administered as soluble proteins. Multimeric antigenic display could markedly improve immunogenicity and should be evaluated in the next generations of vaccines. Here we tested a new virus like particles-based vaccine platform which utilizes the direct fusion of an immunogen of interest to the structural human immunodeficient virus (HIV) protein Gag to increase its surface density and immunogenicity. We compared, in mice, the immunogenicity of RSV-F or hMPV-F based immunogens delivered either as soluble proteins or displayed on the surface of our VLPs. VLP associated F-proteins showed better immunogenicity and induced superior neutralizing responses. Moreover, when combining both VLP associated and soluble immunogens in a heterologous regimen, VLP-associated immunogens provided added benefits when administered as the prime immunization.

Structural characterization of M8C10, a neutralizing antibody targeting a highly conserved prefusion-specific epitope on the metapneumovirus fusion trimerization interface.

IMPORTANCE: Human metapneumovirus (hMPV) is a common pathogen causing lower respiratory tract infections worldwide and can develop severe symptoms in high-risk populations such as infants, the elderly, and immunocompromised patients. There are no approved hMPV vaccines or neutralizing antibodies available for therapeutic or prophylactic use. The trimeric hMPV fusion F protein is the major target of neutralizing antibodies in human sera. Understanding the immune recognition of antibodies to hMPV-F antigen will provide critical insights into developing efficacious hMPV monoclonal antibodies and vaccines.

Exposure to 4,4'-methylene bis (2-chloroaniline) (MbOCA) in New South Wales, Australia.

OBJECTIVES: This study was conducted to determine the level of exposure of 4,4'-methylene bis (2-chloroaniline) (MbOCA) in New South Wales (NSW), Australia. METHODS: An integrated occupational hygiene and biological monitoring program were used to assess the workers' exposure to MbOCA via inhalation, ingestion and dermal contact. This was conducted by personal air monitoring, static air monitoring and surface contamination monitoring of the work environment and biological monitoring of the workers' exposure to MbOCA at nine workplaces in NSW. RESULTS: The air monitoring results for MbOCA gave a geometric mean (GM) of 0.06 µg/m3 and a geometric standard deviation (GSD) of 2.70 and a 95% confidence interval of 0.29 µg/m3. The surface contamination in the main work area showed the highest contamination with a GM of 74 ng/cm2 and a GSD of 17 and a 95% confidence interval of 7,751 ng/cm2. Biological monitoring showed a GM of 0.89 µmol/mol cr and a GSD of 11.9 and a 95% confidence interval of 52 µmol/mol cr. This indicated that 13% of the workers were over the SafeWork NSW Biological Occupational Exposure Limit of 15 µmol/mol cr. CONCLUSIONS: Workers' exposure through inhalation was minimal; however, evidence from biological monitoring of MbOCA suggested that the main contributing factor to exposure was skin absorption. This was attributed to poor housekeeping and inadequate personal protection. Improvements in these areas were recommended, and it was also recommended to improve the awareness of the workers to the adverse effects to their health of exposure to this carcinogen.

Iterative Focused Screening with Biological Fingerprints Identifies Selective Asc-1 Inhibitors Distinct from Traditional High Throughput Screening.

N-methyl-d-aspartate receptors (NMDARs) mediate glutamatergic signaling that is critical to cognitive processes in the central nervous system, and NMDAR hypofunction is thought to contribute to cognitive impairment observed in both schizophrenia and Alzheimer's disease. One approach to enhance the function of NMDAR is to increase the concentration of an NMDAR coagonist, such as glycine or d-serine, in the synaptic cleft. Inhibition of alanine-serine-cysteine transporter-1 (Asc-1), the primary transporter of d-serine, is attractive because the transporter is localized to neurons in brain regions critical to cognitive function, including the hippocampus and cortical layers III and IV, and is colocalized with d-serine and NMDARs. To identify novel Asc-1 inhibitors, two different screening approaches were performed with whole-cell amino acid uptake in heterologous cells stably expressing human Asc-1: (1) a high-throughput screen (HTS) of 3 M compounds measuring 35S l-cysteine uptake into cells attached to scintillation proximity assay beads in a 1536 well format and (2) an iterative focused screen (IFS) of a 45 000 compound diversity set using a 3H d-serine uptake assay with a liquid scintillation plate reader in a 384 well format. Critically important for both screening approaches was the implementation of counter screens to remove nonspecific inhibitors of radioactive amino acid uptake. Furthermore, a 15 000 compound expansion step incorporating both on- and off-target data into chemical and biological fingerprint-based models for selection of additional hits enabled the identification of novel Asc-1-selective chemical matter from the IFS that was not identified in the full-collection HTS.

Brain activity and emotional processing in smokers treated with varenicline.

Prior evidence suggests that varenicline, an effective smoking cessation treatment, may relieve negative affective signs of nicotine withdrawal. We examined varenicline effects on emotional processing in 25 abstinent smokers after 13 days of varenicline and placebo using a within-subject cross-over design. Blood oxygen level dependent (BOLD) functional magnetic resonance imaging was acquired while subjects completed a face emotion identification task. Results showed a significant drug effect, characterized by decreased BOLD signal in dorsal anterior cingulate/medial frontal cortex, occipital cortex and thalamus. Increased BOLD signal was observed in the middle temporal gyrus. Varenicline improved correct response time; however, neither BOLD signal nor performance effects were moderated by emotion type. An exploratory region of interest analysis suggests that varenicline reduced amygdala activity independent of emotional valence. Taken together, these results suggest that observed drug effects on brain activity do not reflect affective changes but rather enhanced early processing of perceptual features of facial stimuli.

The determination of occupational exposure to polycyclic aromatic hydrocarbons by the analysis of 1-hydroxypyrene in urine using a simple automated online column switching device and high-performance liquid chromatography.

An analytical method using a liquid chromatograph combined with a simple online column switching sample pre-treatment system was developed for the determination of 1-hydroxypyrene (1-HP) in urine. This compound is the metabolite of pyrene and is used to assess the exposure of workers to polycyclic aromatic hydrocarbons (PAHs). After enzymatic hydrolysis, a urine sample was directly injected into a high-performance liquid chromatograph (HPLC) where it automatically underwent a sample cleanup using a column switching device. The procedure is simpler than previous methods because it uses only one switching valve, one extraction column and one HPLC pump. The analyte was retained on a short extraction column and after interferences were eluted to waste, was subsequently switched onto the analytical column. This allowed a short analysis time of 15 min. The calibration graph was found to be linear within the concentration range of 0.5 to 20 µg/L with a coefficient of determination exceeding r(2) = 0.99. Recoveries were found to be greater than 96% in the range 1 to 10 µg/L with intermediate precision of 2.5 to 5.8% relative standard deviation. This online method was verified by a comparison with an existing manual method by the analysis of 81 urine samples from workers exposed to PAHs and showed that the test results from both methods were in agreement with a probability obtained from the paired Student's t-test of P > 0.76. The proposed online method was found to be simple, fast and suited to routine analyses of 1-HP in urine for the assessment of occupational exposure to PAHs.