Disruption of Transmembrane Phosphatidylserine Asymmetry by HIV-1 Incorporated SERINC5 Is Not Responsible for Virus Restriction.

Host restriction factor SERINC5 (SER5) incorporates into the HIV-1 membrane and inhibits infectivity by a poorly understood mechanism. Recently, SER5 was found to exhibit scramblase-like activity leading to the externalization of phosphatidylserine (PS) on the viral surface, which has been proposed to be responsible for SER5's antiviral activity. This and other reports that document modulation of HIV-1 infectivity by viral lipid composition prompted us to investigate the role of PS in regulating SER5-mediated HIV-1 restriction. First, we show that the level of SER5 incorporation into virions correlates with an increase in PS levels in the outer leaflet of the viral membrane. We developed an assay to estimate the PS distribution across the viral membrane and found that SER5, but not SER2, which lacks antiviral activity, abrogates PS asymmetry by externalizing this lipid. Second, SER5 incorporation diminished the infectivity of pseudoviruses produced from cells lacking a flippase subunit CDC50a and, therefore, exhibited a higher baseline level of surface-accessible PS. Finally, exogenous manipulation of the viral PS levels utilizing methyl-alpha-cyclodextrin revealed a lack of correlation between external PS and virion infectivity. Taken together, our study implies that the increased PS exposure to SER5-containing virions itself is not directly linked to HIV-1 restriction.

Community perceptions of vaccine advocacy for children under five in rural Guatemala.

Historically, partnerships with community leaders (e.g., religious leaders, teachers) have been critical to building vaccination confidence, but leaders may be increasingly vaccine hesitant. In rural Guatemala, the extent of vaccine hesitancy among community leaders is unclear, as are their perceptions of advocacy for childhood vaccines. We sought to: (i) compare Guatemalan religious leaders' and community leaders' attitudes toward childhood vaccines, (ii) describe leaders' experiences and comfort with vaccination advocacy, and (iii) describe community members' trust in them as vaccination advocates. In 2019, we surveyed religious leaders, other community leaders, and parents of children under five in rural Guatemala. We recorded participant demographic information and assessed participant vaccine hesitancy regarding childhood vaccines. We analyzed data descriptively and via adjusted regression modeling. Our sample included 50 religious leaders, 50 community leaders, and 150 community members (response rate: 99%); 14% of religious leaders and community leaders were vaccine hesitant, similar to community members (P = 0.71). In the prior year, 47% of leaders had spoken about vaccines in their formal role; 85% felt responsible to do so. Only 28% of parents trusted politicians "a lot" for vaccine advice, versus doctors (72%; P < 0.01), nurses (62%; P < 0.01), religious leaders (49%; P < 0.01), and teachers (48%; P < 0.01). In this study, religious leaders and community leaders were willing but incompletely engaged vaccination advocates. Most community members trusted doctors and nurses a lot for vaccination advice; half trusted teachers and religious leaders similarly. Public health officials in rural Guatemala can complement efforts by doctors and nurses through partnerships with teachers and religious leaders to increase vaccination confidence and delivery.

An Examination of Employee Immunization Policies Among Colorado Child Care Providers.

BACKGROUND: Child care employee vaccination policies can protect children and adults from vaccine-preventable diseases (VPDs) in child care programs. We aimed to understand the prevalence and characteristics of employee immunization policies at child care facilities and support among child care administrators for statewide employee vaccination regulations. METHODS: A postal mail survey was distributed to a cross-sectional sample of 300 center-based and 300 home-based child care programs in Colorado. Programs were asked to report whether they had any type of policy requiring employee immunizations and if they would support statewide regulations mandating employee immunizations for influenza, pertussis, and measles. RESULTS: The response rate was 48% (288/600). About 55% of child care programs reported having an employee immunization policy. Child care centers (73%) were more likely than home-based child care programs (30%, P < .001) to report having a policy. Overall, 62% of respondents reported that they would support one or more statewide regulations requiring child care employees to be vaccinated. Home-based programs (71%) were more likely than center-based programs (53%, P = .001) to support one or more statewide child care employee immunization regulations. CONCLUSIONS: Our findings demonstrate the variability of employee immunization policies at child care programs across Colorado. These results may be used to inform strategies to increase employee immunization uptake and reduce the incidence of VPDs, including developing comprehensive employee immunization policies at the facility and state level. Future research is needed to understand vaccination knowledge, attitudes, and behaviors among child care employees including their perspectives on employee vaccination policies and regulations.

Current Challenges and Future Possibilities for Immunization Information Systems.

Immunizations have proven to be an important tool for public health and for reducing the impact of vaccine preventable diseases. To realize the maximum benefits of immunizations, a coordinated effort between public policy, health care providers and health systems is required to increase vaccination coverage and to ensure high-quality data are available to inform clinical and public health interventions. Immunization information systems (IIS) are confidential, population-based, computerized databases that record all immunization doses administered by participating providers to persons residing within a given geopolitical area. The key output of an IIS is high-quality data for use in targeting and monitoring immunization program activities and providing clinical decision support at the point of care. To be truly effective, IISs need to form a nationwide network and repository of immunization data. Since the early 2000s Centers for Disease Control and Prevention has made strides to help IIS move toward a nationwide network through efforts focused on improving infrastructure and functionality, such as the IIS Minimum Functional Standards, and the IIS Annual Report, a self-reported data collection of IIS progress toward achieving the functional standards. While these efforts have helped immunization programs achieve more functional standards, there is a need to shift focus from infrastructure and functionality improvements to high data quality through objective measurement of IIS performance and evaluating critical outcomes. Additionally, realizing the vision of a nationwide repository of high-quality immunization data requires tackling the many challenges that impact data quality and availability including those related to policy, data sharing, data use, aging IIS technology, sustainability, and participation in the IIS. This paper describes the current state of IIS in the United States, critical challenges impacting the quality of data in IIS, and potential components of a future IIS model to address these challenges.

Measurement and improvement as a model to strengthen immunization information systems and overcome data gaps.

BACKGROUND: IIS are important tools in the public health system and exist to improve and protect the nation's health from vaccine-preventable diseases. A network of 62 independent state, territorial, and jurisdictional immunization information systems (IIS) are operated within the United States. These systems are relied upon to implement an increasingly complex vaccination schedule, consolidate and create comprehensive immunization records, as well as monitor vaccine safety, efficacy, and support vaccine delivery. Despite their importance and necessity, the number of varying systems, coupled with jurisdictional policy and resource limitations, presents challenges with standardization, interoperability, data exchange, and the capture of complete immunization records. In partnership with the Centers for Disease Control (CDC) and IIS partners, the American Immunization Registry Association (AIRA) instituted its Measurement and Improvement (M&I) Initiative in 2015 as an innovative effort to evaluate and increase alignment of IIS with national functional standards. Lessons and strategies can be adapted for broader implementation as global systems develop methods to better achieve 2030 Sustainable Development Goal (SDG) targets, particularly related to global population health and infrastructure. METHODS: AIRA works closely with its partners to propose, vet, and refine processes and measures that can be compared across IIS, resulting in a uniform, standardized approach for measurement. The M&I Initiative is conducted as a third-party, independent evaluation through AIRA connecting with IIS pre-production systems to test the IIS response to test messages and measures across multiple content areas prioritized by the IIS community. The process includes three stages: 1) Testing and Discovery, 2) Assessment, and 3) Validation. Content areas currently evaluated include clinical decision support, interoperability transport, HL7 submission/acknowledgement, HL7 message query/response, and data quality. Testing is performed using the AIRA-developed Aggregate Analysis Reporting Tool (AART), an electronic testing tool and user interface specifically designed to compile and visualize results from the measures and tests. RESULTS: The M&I Initiative is voluntary with 86 percent (50/58) of the IIS programs targeted for measurement participating as of Spring 2020. To date, AIRA has actively measured standards alignment and published data on Validation in the first three content areas of Transport, Submission/Acknowledgement, and Query/Response. Thirty-one individual IIS have been validated in at least two of these three content areas. The number of IIS meeting one primary Transport measure has increased from 19 to 39 in three years, an increase of 105 percent. The number of IIS who were able to process the submission of a correctly formatted full immunization record for a patient jumped from 17 to 34, a 200 percent increase from baseline. Similarly, the number of IIS sending standards-conformant HL7 Acknowledgment messages has increased fourteen-fold since measurement began in 2017. The number of IIS who were able to process and respond to a query requesting a patient's evaluated immunization record and forecast increased from nine to 42, a 367 percent increase from baseline. Within the first two quarters of assessment, the percentage of IIS meeting the CDS measures aimed at supporting IIS alignment with ACIP recommendations increased 15 percent from baseline. CONCLUSION: The M&I initiative has helped to reduce variability across IIS and strengthen immunization data in IIS that is more complete, accurate, and can be utilized with confidence. The successes and experience offer an innovative model that could be adapted to standardize measures of success and data-sharing capabilities across global borders, particularly of value in achieving SDGs aimed at ensuring healthy lives and promoting well-being for all ages through strengthened immunization systems.

Adapting and piloting a vaccine hesitancy questionnaire in rural Guatemala.

INTRODUCTION: We sought to (i) adapt a Spanish-language vaccine hesitancy (VH) tool to rural Guatemala, (ii) pilot the tool with 150 parents of children ≤ 5 years, and (iii) measure if parent scores associated with child under-vaccination. METHODS: We used implementation science to develop the adapted Guatemalan Vaccine Attitudes (GuaVA) tool, piloting it with 150 parents of children ≤ 5 years, and performing descriptive and adjusted regression analyses. RESULTS: Of 150 parents (response rate 99%), 55% (n = 83) of parents expressed a degree of VH. Children of parents with highly hesitant scores (n = 22) had 2.5 times the odds (OR 2.5; 95% CI: 1.2, 5.4) of being undervaccinated at 19 months, referent children of non-hesitant parents (n = 67). CONCLUSIONS: Vaccine hesitancy may be more prevalent in rural Guatemala than suspected. Implementation science facilitated the adaptation of a VH tool to rural Guatemala and may assist investigators in other settings.

Design of Next Generation Polymyxins with Lower Toxicity: The Discovery of SPR206.

Polymyxins are an important class of antibiotics for the treatment of bacterial infections due to multidrug resistant Gram-negative pathogens. However, their clinical utility is limited by nephrotoxicity. Here, we report a series of promising next generation polymyxin nonapeptides identified on the basis of our understanding of the relationship of structure with activity, cytotoxicity, and kidney compartment accumulation. We demonstrate that nonapeptides with an amine-containing N-terminal moiety of specific regio- and stereochemistry possess superior in vitro activity, together with lower cytotoxicity compared to polymyxin B. We further demonstrate that compounds with a ß-branched aminobutyrate N-terminus with an aryl substituent offer a promising combination of low cytotoxicity and kidney exposure, leading to low toxicity in the mouse. From this series, SPR206 has been selected as a development candidate.

Anti-Inflammatory Benefits of Antibiotics: Tylvalosin Induces Apoptosis of Porcine Neutrophils and Macrophages, Promotes Efferocytosis, and Inhibits Pro-Inflammatory CXCL-8, IL1α, and LTB4 Production, While Inducing the Release of Pro-Resolving Lipoxin A4 and Resolvin D1.

Excessive accumulation of neutrophils and their uncontrolled death by necrosis at the site of inflammation exacerbates inflammatory responses and leads to self-amplifying tissue injury and loss of organ function, as exemplified in a variety of respiratory diseases. In homeostasis, neutrophils are inactivated by apoptosis, and non phlogistically removed by neighboring macrophages in a process known as efferocytosis, which promotes the resolution of inflammation. The present study assessed the potential anti-inflammatory and pro-resolution benefits of tylvalosin, a recently developed broad-spectrum veterinary macrolide derived from tylosin. Recent findings indicate that tylvalosin may modulate inflammation by suppressing NF-κB activation. Neutrophils and monocyte-derived macrophages were isolated from fresh blood samples obtained from 12- to 22-week-old pigs. Leukocytes exposed to vehicle or to tylvalosin (0.1, 1.0, or 10 µg/mL; 0.096-9.6 µM) were assessed at various time points for apoptosis, necrosis, efferocytosis, and changes in the production of cytokines and lipid mediators. The findings indicate that tylvalosin increases porcine neutrophil and macrophage apoptosis in a concentration- and time-dependent manner, without altering levels of necrosis or reactive oxygen species production. Importantly, tylvalosin increased the release of pro-resolving Lipoxin A4 (LXA4) and Resolvin D1 (RvD 1 ) while inhibiting the production of pro-inflammatory Leukotriene B4 (LTB4) in Ca2+ ionophore-stimulated porcine neutrophils. Tylvalosin increased neutrophil phospholipase C activity, an enzyme involved in releasing arachidonic acid from membrane stores. Tylvalosin also inhibited pro-inflammatory chemokine (C-X-C motif) ligand 8 (CXCL-8, also known as Interleukin-8) and interleukin-1 alpha (IL-1α) protein secretion in bacterial lipopolysaccharide-stimulated macrophages. Together, these data illustrate that tylvalosin has potent immunomodulatory effects in porcine leukocytes in addition to its antimicrobial properties.

Seeding Australian Regulation of Genomics in the Cloud ­ Elizabeth Abbott and Thomas Faunce.

Cloud computing has facilitated a revolution in genome sequencing. As big data and personalised medicine increase in popularity in Australia, are the legal and regulatory regimes surrounding this nascent area of scientific research and clinical practice able to protect this private information? An examination of the current regulatory regime in Australia, including the Privacy Act 1988 (Cth) and medical research laws that govern cloud-based genomics research highlights that the key challenge of such research is to protect the interests of participants while also promoting collaborative research processes. This examination also highlights the potential effect that the Trans-Pacific Partnership Agreement's Electronic Commerce Chapter may have had on using the cloud for genomics and what the consequences may have been for researchers, clinicians and individuals. Lessons learnt here will be relevant to studying similar impacts from other trade and investment agreements such as the Trade in Services Agreement (TiSA).

RSV604, a novel inhibitor of respiratory syncytial virus replication.

Respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infections worldwide, yet no effective vaccine or antiviral treatment is available. Here we report the discovery and initial development of RSV604, a novel benzodiazepine with submicromolar anti-RSV activity. It proved to be equipotent against all clinical isolates tested of both the A and B subtypes of the virus. The compound has a low rate of in vitro resistance development. Sequencing revealed that the resistant virus had mutations within the nucleocapsid protein. This is a novel mechanism of action for anti-RSV compounds. In a three-dimensional human airway epithelial cell model, RSV604 was able to pass from the basolateral side of the epithelium effectively to inhibit virus replication after mucosal inoculation. RSV604, which is currently in phase II clinical trials, represents the first in a new class of RSV inhibitors and may have significant potential for the effective treatment of RSV disease.

A comparison of health-care costs involved in treating people with and without Parkinson's disease in Southern Sydney, New South Wales, Australia.

Twelve community-dwelling subjects with Parkinson's disease (PD) and 12 age-matched healthy controls completed a 3-month diary of health-care utilisation in Sydney, Australia. The mean age for the PD group was 71.3 years (SD 5.9, range 62-82 years) versus 73.2 years (SD 6.7, range 63-83 years) for the control group. The mean disease duration of the PD group was 6.8 years (SD 3.6, range 2-14 years). The median Hoehn and Yahr stage was 3 (range 1-3). The mean 3-month total (both related and 'unrelated' to PD) health-care cost for the PD group was significantly higher than that for the 'healthy' control group (1,755 Australian dollars, SD 1,201 versus 413 Australian dollars, SD 515, P=0.001). Medication was the most costly component for both groups (PD 636 Australian dollars, SD 226 versus controls 175 Australian dollars, SD 233, P<0.001) followed by general practitioner or specialist medical expenses (PD 564 Australian dollars, SD 670 versus controls 205 Australian dollars, SD 397, p=0.12) and allied health-care costs (PD 323 Australian dollars, SD 178 versus controls 21 Australian dollars, SD 43, p<0.001). In the PD subgroup, the health-care costs attributed to PD during the 3-month period were significantly higher than health-care costs 'unrelated' to PD (1,202 Australian dollars, SD 820 versus 553 Australian dollars, SD 591, p=0.03). On subgroup analysis, allied health-care costs (related to PD) achieved statistical significance (304 Australian dollars, SD 180 versus 19 Australian dollars, SD 19, p<0.0001), whereas medication and general practitioner or specialist costs did not. In conclusion, we found that the total direct health-care cost of PD for patients with Hoehn and Yahr stage 3 was four times that of age- and sex-matched 'healthy' controls. The estimated annual cost (7,020 Australian dollars per patient) in our patient cohort was comparable to that reported in the United States and Europe.

Pharmacokinetics, safety, and efficacy of a liposome encapsulated thymidylate synthase inhibitor, OSI-7904L [(S)-2-[5-[(1,2-dihydro-3-methyl-1-oxobenzo[f]quinazolin-9-yl)methyl]amino-1-oxo-2-isoindolynl]-glutaric acid] in mice.

OSI-7904L [(S)-2-[5-[(1,2-dihydro-3-methyl-1-oxobenzo[f]quinazolin-9-yl)methyl]amino-1-oxo-2-isoindolynl]-glutaric acid] is a liposomal formulation of the highly specific, noncompetitive, thymidylate synthase inhibitor OSI-7904 (also known as GW1843, 1843U89, and GS7904). The liposome formulation was developed to enhance the therapeutic index and dose schedule convenience of this potent antifolate compound. The studies presented here were conducted to determine the antitumor efficacy, distribution, pharmacokinetics, and safety of OSI-7904L in mice. In a human colon adenocarcinoma xenograft model in mice, OSI-7904L demonstrated superior antitumor efficacy compared with OSI-7904 or 5-fluorouracil. Furthermore, OSI-7904L could be administered less frequently than OSI-7904 although still generating greater tumor growth inhibition. Distribution studies confirmed that OSI-7904L-treated animals had much greater plasma, tissue, and tumor exposure than did OSI-7904-treated animals. Tumor exposures, based on area under the curve, in OSI-7904L-treated mice were increased over 100-fold compared with tumor exposures in OSI-7904-treated mice. Plasma exposures following OSI-7904L administration were greater than dose proportional consistent with saturation of plasma clearance mechanisms. OSI-7904L was much more toxic than OSI-7904 in the mouse with primary toxicities to the intestines, bone marrow, and thymus. Minimal toxicity to the lungs and liver was noted. These data clearly demonstrated that in mice, OSI-7904L has an increased plasma residence time as well as increased tissue and tumor exposure compared with OSI-7904, thus resulting in increased potency and toxicity. Potential benefits of OSI-7904L include improved efficacy and a more convenient schedule of administration.