Phylogenetic distribution and membrane topology of the LytR-CpsA-Psr protein family

BACKGROUND: The bacterial cell wall is the target of many antibiotics and cell envelope constituents are critical to host-pathogen interactions. To combat resistance development and virulence, a detailed knowledge of the individual factors involved is essential. Members of the LytR-CpsA-Psr family of cell envelope-associated attenuators are relevant for beta-lactam resistance, biofilm formation, and stress tolerance, and they are suggested to play a role in cell wall maintenance. However, their precise function is still unknown. This study addresses the occurrence as well as sequence-based characteristics of the LytR-CpsA-Psr proteins. RESULTS: A comprehensive list of LytR-CpsA-Psr proteins was established, and their phylogenetic distribution and clustering into subgroups was determined. LytR-CpsA-Psr proteins were present in all Gram-positive organisms, except for the cell wall-deficient Mollicutes and one strain of the Clostridiales. In contrast, the majority of Gram-negatives did not contain LytR-CpsA-Psr family members. Despite high sequence divergence, the LytR-CpsA-Psr domains of different subclusters shared a highly similar, predicted mixed alpha/beta-structure, and conserved charged residues. PhoA fusion experiments, using MsrR of Staphylococcus aureus, confirmed membrane topology predictions and extracellular location of its LytR-CpsA-Psr domain. CONCLUSIONS: The LytR-CpsA-Psr domain is unique to bacteria. The presence of diverse subgroups within the LytR-CpsA-Psr family might indicate functional differences, and could explain variations in phenotypes of respective mutants reported. The identified conserved structural elements and amino acids are likely to be important for the function of the domain and will help to guide future studies of the LytR-CpsA-Psr proteins

Detection of High Titers of Antibody against Helicobacter Cysteine-Rich Proteins A, B, C, and E in Helicobacter pylori-Infected Individuals

The family of Helicobacter cysteine-rich proteins (Hcp) constitutes one of the largest protein families that are specific for proteobacteria from the delta/epsilon subgroup. Most of the proteins belonging to this family have so far only been recognized on the genome level. To investigate the expression of Hcp proteins in vivo we analyzed titers of antibody against HcpA (HP0211), HcpB (HP0336), HcpC (HP1098), and HcpE (HP0235) in sera from 30 Helicobacter pylori-positive individuals and in a control group of six H. pylori-negative individuals. Significantly higher titers of antibody were observed for H. pylori-positive individuals (P < 0.00005). The highest and lowest titers were observed for HcpC (Δ mean = 1.06) and HcpB (Δ mean = 0.333), respectively. There is a clear correlation among anti-HcpA, -HcpC, and -HcpE immunoglobulin G titers in H. pylori-positive individuals (correlation > 0.7), but there is only a weak correlation for HcpB (correlation < 0.4). These results confirm that Hcp proteins are expressed by H. pylori under natural environmental conditions and that these proteins are recognized by the immune system of the host. The observed correlations are in agreement with the expected distribution of Hcp proteins among H. pylori strains. HcpA, HcpC, and HcpE are present in the genomes of strains 26695 and J99, whereas HcpB is absent from most strains. Since Hcp proteins are specific for H. pylori, immunological assays including Hcp proteins might be of value to detect H. pylori infection and perhaps to distinguish among different groups of H. pylori-positive patients

Health Care Professionals’ Interest in Vaccination Training in Switzerland: A Quantitative Survey

Objectives: Health care professionals (HCPs) play an important role for patients’ vaccination decisions. To counsel patients/clients appropriately, HCPs need current factual knowledge about vaccines and strong communication skills. Methods: We conducted an online survey with physicians, pharmacists, nurses, and midwives in Switzerland (01.11.2020–31.03.2021). We evaluated: 1) interest in vaccination knowledge and counseling training; 2) vaccination recommendation practices; 3) experience with vaccination counseling/administration; 4) comfort level in addressing vaccine hesitancy (VH); 5) perspectives on patient/client VH, delays, and refusals. Results: In total, 1,933 practicing HCPs responded (496 physicians, 226 pharmacists, 607 nurses, 604 midwives). 43% physicians, 31% pharmacists, 15% nurses, and 23% midwives felt comfortable counseling VH patients/clients. 96% physicians, 98% pharmacists, 85% nurses, and 91% midwives were interested in additional vaccination-related training. All professionals mentioned safety, efficacy, and side effects as topics of most interest for additional training. Conclusion: Results demonstrate a high interest among HCPs for additional vaccination-related training. In addition to factual information about vaccination, such training will likely benefit from a communication component, given the low rates of comfort reported by HCPs when counseling VH patients/clients. </p

Healthcare professional and professional stakeholders’ perspectives on vaccine mandates in switzerland: a mixed-methods study

BACKGROUND: There currently are no mandatory vaccines in Switzerland. However, Swiss federal legislation allows for vaccination mandates in settings where the risk of transmission to vulnerable groups is high, such as healthcare professionals (HCPs) working with vulnerable patients. Since HCPs are trusted information sources, a priority population for COVID-19 vaccination, and potentially subjected to mandates, we investigated HCP perspectives on mandates. METHODS: A national online survey was administered to HCPs (October 2020-March 2021), including vaccine mandates questions concerning patients (measles) and HCPs (influenza). We qualitatively investigated HCP mandate perspectives through: (1) 34 interviews with HCPs, HCP professional society representatives, and health authorities; (2) a focus group discussion (FGD) with complementary medicine (CM) and biomedical physicians, and Swiss Federal Vaccination Commission members. RESULTS: 1933 participants (496 physicians, 226 pharmacists, 607 nurses, 604 midwives) responded to the survey. Quantitative results show all professional groups preferred shared parent-HCP measles vaccine decisions (65%, 54%, 50%, 48%, respectively; p for trend < 0.001). Midwives (87%) and nurses (70%) preferred individual influenza vaccination decisions for HCPs, while physicians (49%) and pharmacists (44%) preferred shared employee-employer decisions (p for trend < 0.001). Physicians (p < .001) and pharmacists (p < .01) with CM training favored individual influenza vaccination decisions. Qualitative results show general HCP opposition to vaccine mandates, mainly because participants argued how other approaches, such as HCP training and better information, could encourage uptake. Arguments against COVID-19 mandates included insufficiently documented long-term safety/efficacy data. From participants’ perspectives, mandated vaccination should be used as a last resort. Some participants expressed fear that with mandates, notably for influenza and COVID-19, some HCPs might leave their jobs. HCPs were unsure what vaccine mandates would concretely look like in practice, particularly regarding sanctions for non-compliance and enforcement. CONCLUSION: In Switzerland, HCPs generally were opposed to vaccination mandates. Clarity and guidance are needed from health authorities to better inform discussions around vaccine mandates