Factors Associated with Vaccine Hesitancy in the State of Alaska.

BACKGROUND: Vaccine hesitancy (VH) is a major public health problem which includes not only concerns about vaccine, but often includes beliefs not aligned with evidence. The etiology of VH is complicate, including genuine concerns about vaccine safety to beliefs in disproven or unsubstantiated theories. Understanding VH includes determining the factors associated with people most likely to be susceptible. OBJECTIVES: To identify characteristics of persons more likely to have VH based on perceived vaccine safety. DESIGN: We conducted a cross sectional, targeted online survey of 1,024 respondents. METHODS: The survey collected data on demographics and vaccine beliefs. A seven-question VH scale was created based on an extensive literature review. When measured, this scale had high internal validity based on Cronbach's Alpha (α = 0.80, 95% CI, 0.79, 0.82) and provided a continuous measurement to understand the VH degree. RESULTS: Persons reporting a conservative political ideology had the highest VH score ([Formula: see text] = 15.0), followed by other ([Formula: see text] =12.7), moderate ([Formula: see text] =9.29), and liberal ([Formula: see text] =6.72). Education level was another strong indicator of VH, decreasing from the highest score of high school graduates ( [Formula: see text] = 13.2) to graduate degree holders ([Formula: see text] = 9.22). CONCLUSION: The more conservative a person's political ideology, the higher the VHS. We asked for ideology, rather than political party, although they are related, they are not the same. Ideology influences more domains than voting patterns. Higher educational attainment shows a protective effect against VH. These findings have direct implication for public health interventions, new interventions should be designed to be more accessible for persons with less formal education and more amenable to persons who hold a more conservative ideology.

Topographic diversity of fungal and bacterial communities in human skin.

Traditional culture-based methods have incompletely defined the microbial landscape of common recalcitrant human fungal skin diseases, including athlete's foot and toenail infections. Skin protects humans from invasion by pathogenic microorganisms and provides a home for diverse commensal microbiota. Bacterial genomic sequence data have generated novel hypotheses about species and community structures underlying human disorders. However, microbial diversity is not limited to bacteria; microorganisms such as fungi also have major roles in microbial community stability, human health and disease. Genomic methodologies to identify fungal species and communities have been limited compared with those that are available for bacteria. Fungal evolution can be reconstructed with phylogenetic markers, including ribosomal RNA gene regions and other highly conserved genes. Here we sequenced and analysed fungal communities of 14 skin sites in 10 healthy adults. Eleven core-body and arm sites were dominated by fungi of the genus Malassezia, with only species-level classifications revealing fungal-community composition differences between sites. By contrast, three foot sites--plantar heel, toenail and toe web--showed high fungal diversity. Concurrent analysis of bacterial and fungal communities demonstrated that physiologic attributes and topography of skin differentially shape these two microbial communities. These results provide a framework for future investigation of the contribution of interactions between pathogenic and commensal fungal and bacterial communities to the maintainenace of human health and to disease pathogenesis.

Kinetic analysis of PCNA clamp binding and release in the clamp loading reaction catalyzed by Saccharomyces cerevisiae replication factor C.

DNA polymerases require a sliding clamp to achieve processive DNA synthesis. The toroidal clamps are loaded onto DNA by clamp loaders, members of the AAA+family of ATPases. These enzymes utilize the energy of ATP binding and hydrolysis to perform a variety of cellular functions. In this study, a clamp loader-clamp binding assay was developed to measure the rates of ATP-dependent clamp binding and ATP-hydrolysis-dependent clamp release for the Saccharomyces cerevisiae clamp loader (RFC) and clamp (PCNA). Pre-steady-state kinetics of PCNA binding showed that although ATP binding to RFC increases affinity for PCNA, ATP binding rates and ATP-dependent conformational changes in RFC are fast relative to PCNA binding rates. Interestingly, RFC binds PCNA faster than the Escherichia coli γ complex clamp loader binds the ß-clamp. In the process of loading clamps on DNA, RFC maintains contact with PCNA while PCNA closes, as the observed rate of PCNA closing is faster than the rate of PCNA release, precluding the possibility of an open clamp dissociating from DNA. Rates of clamp closing and release are not dependent on the rate of the DNA binding step and are also slower than reported rates of ATP hydrolysis, showing that these rates reflect unique intramolecular reaction steps in the clamp loading pathway.

Vaccine safety beliefs in the state of Alaska.

Objectives: Identifying the key factors associated with vaccine hesitancy remains a challenge as has been highlighted throughout the COVID-19 vaccine roll out and pandemic. The aim of this study was to determine characteristics associated with vaccine safety and compare perceived safety by vaccine. Our hypothesis is that vaccine safety perception will vary by vaccine with COVID-19 as ranked lowest for safety. Study design: Cross sectional. Methods: A statewide sample (n = 1024) responded to an online 28-point questionnaire via anonymous linked invitation. Results: Among the eight vaccines assessed, COVID-19 had the lowest perceived safety (53.13%) followed by human papillomavirus HPV (63.38%). A binomial logistic regression assessed COVID-19 vaccine safety beliefs (safe v not safe) finding age, political orientation, and perceived safety of certain vaccines as statistically significant. As age increased by year, vaccine safety beliefs increased. Persons who identified as conservative demonstrated less belief in vaccine safety than all other groups. Among persons who did not perceive the COVID-19 vaccine as safe, 65.8% believed chicken pox was safe, 63.3% and 61.1% perceived hepatitis A& B were safe. Conclusions: These findings demonstrate that vaccine safety beliefs differ by vaccine and that persons who do not believe in the safety of the COVID-19 are not exclusively against all vaccines. Understanding factors that increase vaccine safety by vaccine could assist in developing an intervention which could increase belief in safety for all vaccines.

Pan-PCR, a computational method for designing bacterium-typing assays based on whole-genome sequence data.

With increasing rates of antibiotic resistance, bacterial infections have become more difficult to treat, elevating the importance of surveillance and prevention. Effective surveillance relies on the availability of rapid, cost-effective, and informative typing methods to monitor bacterial isolates. PCR-based typing assays are fast and inexpensive, but their utility is limited by the lack of targets which are capable of distinguishing between strains within a species. To identify highly informative PCR targets from the growing base of publicly available bacterial genome sequences, we developed pan-PCR. This computer algorithm uses existing genome sequences for isolates of a species of interest and identifies a set of genes whose patterns of presence or absence provide the best discrimination between strains in this species. A set of PCR primers targeting the identified genes is then designed, with each PCR product being of a different size to allow multiplexing. These target DNA regions and PCR primers can then be utilized to type bacterial isolates. To evaluate pan-PCR, we designed an assay for the emerging pathogen Acinetobacter baumannii. Taking as input a set of 29 previously sequenced genomes, pan-PCR identified 6 genetic loci whose presence or absence was capable of distinguishing all the input strains. This assay was applied to a set of patient isolates, and its discriminatory power was compared to that of multilocus sequence typing (MLST) and whole-genome optical maps. We found that the pan-PCR assay was capable of making clinically relevant distinctions between strains with identical MLST profiles and showed a discriminatory power similar to that of optical maps. Pan-PCR represents a tool capable of exploiting available genome sequence data to design highly discriminatory PCR assays. The ease of design and implementation makes this approach feasible for diagnostic facilities of all sizes.

Mass spectrometric characterization and activity of zinc-activated proinsulin C-peptide and C-peptide mutants.

Numerous reports have demonstrated an active role for proinsulin C-peptide in ameliorating chronic complications associated with diabetes mellitus. It has been recently reported that some of these activities are dependent upon activation of C-peptide with certain metal ions, such as Fe(II), Cr(III) or Zn(II). In an effort to gain a greater understanding of the structure/function dependence of the peptide-metal interactions responsible for this activity, a series of experiments involving the use of electrospray ionization (ESI), matrix assisted laser desorption/ionization (MALDI) and collision-induced dissociation-tandem mass spectrometry (CID-MS/MS) of C-peptide in the presence or absence of Zn(II) have been carried out. Additionally, various C-peptide mutants with alanine substitution at individual aspartic acid or glutamic acid residues throughout the C-peptide sequence were analyzed. CID-MS/MS of wild type C-peptide in the presence of Zn(II) indicated multiple sites for metal binding, localized at acidic residues within the peptide sequence. Mutations of individual acidic residues did not significantly affect this fragmentation behavior, suggesting that no single acidic residue is critical for binding. However, ESI-MS analysis revealed an approximately 50% decrease in relative Zn(II) binding for each of the mutants compared to the wild type sequence. Furthermore, a significant decrease in activity was observed for each of the Zn(II)-activated mutant peptides compared to the wild type C-peptide, indicated by measurement of ATP released from erythrocytes, with a 75% decrease observed for the Glu27 mutant. Additional studies on the C-terminal pentapeptide of C-peptide EGSLQ, as well as a mutant C-terminal pentapeptide sequence AGSLQ, revealed that substitution of the glutamic acid residue resulted in a complete loss of activity, implicating a central role for Glu27 in Zn(II)-mediated C-peptide activity.

A microfluidic technique for monitoring bloodstream analytes indicative of C-peptide resistance in type 2 diabetes.

A simple poly(dimethylsiloxane) (PDMS) microchip was employed to establish a relationship between red blood cell (RBC) antioxidant status and the ability of RBCs to interact with metal-activated C-peptide, a bio-active peptide reported to reduce some complications often associated with diabetes. It is known that the reduced form of glutathione (GSH) levels in the RBCs obtained from people with type 2 diabetes are lower in comparison to those RBCs obtained from healthy controls and accordingly, this correlation has the potential to implicate type 2 diabetes in high-risk individuals. A parallel channel microfluidic device for the quantification of GSH in age-based fractions, along with control and diabetic RBCs is described. Important to the fluorescence-based measurement is the simultaneous determination of the antioxidant without prior separation in either a six- or twelve-channel microchip. Here, we separated the RBCs using a density-based Percoll solution and quantitatively determined the concentration of GSH in younger, less dense RBCs to be increased more than 2-fold (336.7 +/- 29.6 amol/RBC) than older, more dense RBCs (137.0 +/- 25.3 amol/RBC). The ability of C-peptide to interact with the RBC membrane of the separated fractions was determined by immunoassay and it was found that the recovery of the C-peptide added to the younger RBCs increased by more than 40.6 +/- 12.7% above basal levels while with the older cells C-peptide increased by only 9.18 +/- 4.60%. These results suggest that GSH concentrations in the RBC may be useful in screening for resistance to C-peptide in vivo.

Structural determination of the principal byproduct of the lithium-ammonia reduction method of methamphetamine manufacture.

One common method of illicit methamphetamine manufacture utilizes an alkali metal, typically lithium, and liquid ammonia to chemically reduce ephedrine or pseudoephedrine to form methamphetamine. This method is often referred to as the lithium-ammonia reduction method or the Birch reduction method. While the hydroxyl group of ephedrine is more reactive than the aromatic ring, excess alkali metal and the presence of a proton source allow the formation of a cyclohexadiene byproduct not found in samples of methamphetamine produced from other manufacturing methods. A sample enriched in this byproduct was generated and characterized using nuclear magnetic resonance (NMR) spectroscopy, gas chromatography-mass spectrometry (GC-MS), infrared (IR) spectrophotometry, and ultraviolet (UV) spectrophotometry. The chemical structure of this byproduct was determined to be 1-(1',4'-cyclohexadienyl)-2-methylaminopropane (CMP).

Suzuki-Miyaura cross-coupling of 3-pyridyl triflates with 1-alkenyl-2-pinacol boronates.

Palladium-catalyzed Suzuki-type couplings of 3-pyridyl triflates with alkenyl pinacol boronates proceed in good to excellent yield. Optimized conditions use Pd(PPh(3))(4) (10 mol %) as catalyst with K(3)PO(4) (3 equiv) as base in dioxane.

Zinc-activated C-peptide resistance to the type 2 diabetic erythrocyte is associated with hyperglycemia-induced phosphatidylserine externalization and reversed by metformin.

Insulin resistance can broadly be defined as the diminished ability of cells to respond to the action of insulin in transporting glucose from the bloodstream into cells and tissues. Here, we report that erythrocytes (ERYs) obtained from type 2 diabetic rats display an apparent resistance to Zn(2+)-activated C-peptide. Thus, the aims of this study were to demonstrate that Zn(2+)-activated C-peptide exerts potentially beneficial effects on healthy ERYs and that these same effects on type 2 diabetic ERYs are enhanced in the presence of metformin. Incubation of ERYs (obtained from type 2 diabetic BBZDR/Wor-rats) with Zn(2+)-activated C-peptide followed by chemiluminescence measurements of ATP resulted in a 31.2 +/- 4.0% increase in ATP release from these ERYs compared to a 78.4 +/- 4.9% increase from control ERYs. Glucose accumulation in diabetic ERYs, measured by scintillation counting of (14)C-labeled glucose, increased by 35.8 +/- 1.3% in the presence of the Zn(2+)-activated C-peptide, a value significantly lower than results obtained from control ERYs (64.3 +/- 5.1%). When Zn(2+)-activated C-peptide was exogenously added to diabetic ERYs, immunoassays revealed a 32.5 +/- 8.2% increase in C-peptide absorbance compared to a 64.4 +/- 10.3% increase in control ERYs. Phosphatidylserine (PS) externalization and metformin sensitization of Zn(2+)-activated C-peptide were examined spectrofluorometrically by measuring the binding of FITC-labeled annexin to PS. The incubation of diabetic ERYs with metformin prior to the addition of Zn(2+)-activated C-peptide resulted in values that were statistically equivalent to those of controls. Summarily, data obtained here demonstrate an apparent resistance to Zn(2+)-activated C-peptide by the ERY that is corrected by metformin.

Conversion of epoxides to 1,3-dioxolanes catalyzed by tin(II) chloride.

Anhydrous tin(II) chloride is an efficient catalyst for the reaction of epoxides with acetone to prepare 2,2-dimethyl-1,3-dioxolanes (acetonides) in good to excellent yields. Mono-, di-, and trisubstituted epoxides participate equally well in this diastereospecific reaction. The use of single enantiomer epoxides under the reported conditions results in significant erosion of optical activity.