Alcohol Consumption Among Cannabis Users in Vermont

Objective: To examine the relationship between cannabis and alcohol consumption in Vermont; after the legalization of cannabis in 2018. Methods: We used Vermont\u27s Behavioral Risk Factor Surveillance System (BRFSS) survey; this data included only participants who had reported alcohol use in the past year (n = 3,781). We used multiple linear regression to compare frequency of cannabis use against days per month of alcohol consumption and binomial logistic to compare regression to the frequency in which cannabis was used against the odds of Heavy Alcohol Consumption (HAC). Results: We found that low and moderate cannabis users had more days of alcohol consumption on average than non-users. High cannabis users, however, had fewer days on average than non-users of cannabis. Additionally, compared to non-users, low, moderate, and high cannabis use was associated with increased odds of HAC. Conclusion: Our results suggest that cannabis use may be predictive of alcohol consumption, so individuals, clinicians, and policymakers should be aware of how cannabis use could impact a person\u27s overall health picture

Pre-Clinical Evaluation of a Novel Nanoemulsion-Based Hepatitis B Mucosal Vaccine

Hepatitis B virus infection remains an important global health concern despite the availability of safe and effective prophylactic vaccines. Limitations to these vaccines include requirement for refrigeration and three immunizations thereby restricting use in the developing world. A new nasal hepatitis B vaccine composed of recombinant hepatitis B surface antigen (HBsAg) in a novel nanoemulsion (NE) adjuvant (HBsAg-NE) could be effective with fewer administrations.Physical characterization indicated that HBsAg-NE consists of uniform lipid droplets (349+/-17 nm) associated with HBsAg through electrostatic and hydrophobic interactions. Immunogenicity of HBsAg-NE vaccine was evaluated in mice, rats and guinea pigs. Animals immunized intranasally developed robust and sustained systemic IgG, mucosal IgA and strong antigen-specific cellular immune responses. Serum IgG reached > or = 10(6) titers and was comparable to intramuscular vaccination with alum-adjuvanted vaccine (HBsAg-Alu). Normalization showed that HBsAg-NE vaccination correlates with a protective immunity equivalent or greater than 1000 IU/ml. Th1 polarized immune response was indicated by IFN-gamma and TNF-alpha cytokine production and elevated levels of IgG(2) subclass of HBsAg-specific antibodies. The vaccine retains full immunogenicity for a year at 4 degrees C, 6 months at 25 degrees C and 6 weeks at 40 degrees C. Comprehensive pre-clinical toxicology evaluation demonstrated that HBsAg-NE vaccine is safe and well tolerated in multiple animal models.Our results suggest that needle-free nasal immunization with HBsAg-NE could be a safe and effective hepatitis B vaccine, or provide an alternative booster administration for the parenteral hepatitis B vaccines. This vaccine induces a Th1 associated cellular immunity and also may provide therapeutic benefit to patients with chronic hepatitis B infection who lack cellular immune responses to adequately control viral replication. Long-term stability of this vaccine formulation at elevated temperatures suggests a direct advantage in the field, since potential excursions from cold chain maintenance could be tolerated without a loss in therapeutic efficacy

Impact of an N‑terminal Polyhistidine Tag on Protein Thermal Stability

For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic properties. In this study, we determine the effect of N-terminal His-tags on the thermal stability of select proteins using differential scanning fluorimetry and identify that the removal of the His-tag can have both beneficial and deleterious effects on their stability

Characterization of Stability and Nasal Delivery Systems for Immunization with Nanoemulsion-Based Vaccines

Abstract Background: Many infectious diseases that cause significant morbidity and mortality, especially in the developing world, could be preventable through vaccination. The effort to produce safe, thermally stable, and needle-free mucosal vaccines has become increasingly important for global health considerations. We have previously demonstrated that a thermally stable nanoemulsion, a mucosal adjuvant for needle-free nasal immunization, is safe and induces protective immunity with a variety of antigens, including recombinant protein. The successful use of nanoemulsion-based vaccines, however, poses numerous challenges. Among the challenges is optimization of the formulation to maintain thermal stability and potency and another is accuracy and efficiency of dispensing the vaccines to the nasal mucosa in the anterior and turbinate region of the nasal cavity or potentially to the nasopharynx-associated lymphoid tissue. Methods: We have examined the effects of different diluents [phosphate-buffered saline (PBS) and 0.9% NaCl] on the stability and potency of nanoemulsion-based vaccines. In addition, we have determined the efficiency of delivering them using commercially available nasal spray devices (Pfeiffer SAP-62602 multidose pump and the BD Hypak SCF 0.5?ml unit dose AccusprayTM). Results: We report the stability and potency of PBS?diluted ovalbumin?nanomeulsion mixtures for up to 8 months and NaCl-diluted mixtures up to 6 months when stored at room temperature. Significant differences in spray characteristics including droplet size, spray angle, plume width, and ovality ratios were observed between the two pumps. Further, we have demonstrated that the nanoemulsion-based vaccines are not physically or chemically altered and retain potency following actuation with nasal spray devices. Using either device, the measured spray characteristics suggest deposition of nanoemulsion-based vaccines in inductive tissues located in the anterior region of the nasal cavity. Conclusions: The results of this study suggest that nanoemulsion-based vaccines do not require specially engineered delivery devices and support their potential use as nasopharyngeal vaccine adjuvants.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/85139/1/jamp_2009_0766.pd

In vitro analyses of HBsAg-NE stability.

<p>The comparison of HBsAg-NE stored at test temperature conditions to freshly prepared formulation using SDS-PAGE followed by silver staining (S) or Western immunoblotting (W) is shown. Lanes are labeled according to sample storage conditions as follows- 1: fresh, 2: 4°C, 3: 25°C and 4: 40°C. Samples were stored for (A) 6 weeks, (B) 6 months (24 weeks), or (C) 1 year (52 weeks) at the three test temperatures. Each lane contains 0.5 μg of antigen. Arrow indicates the HBsAg major band. (D) Particle size comparison of NE alone to freshly mixed HBsAg-NE and formulation stored up to a year. Mean diameter of particles is shown in microns +/− SD.</p

Stability of HBsAg.

<p>Silver stained SDS-PAGE: Analysis of freshly prepared vs. 72 hour incubated HBs antigen mixed with 20% NE (A). Conditions of the incubation are specified above the gel. HBsAg monomer and dimer are indicated with arrows at 24 kDa and 48 kDa bands. Western blot: The protein configuration is identical to as described for SDS-PAGE (B). HBsAg is detected using a polyclonal anti-HBsAg antibody for all incubation conditions.</p