Stakeholder engagement to ensure the sustainability of biobanks: a survey of potential users of biobank services

Biobanks are important infrastructures facilitating biomedical research. After a decade of rolling out such infrastructures, a shift in attention to the sustainability of biobanks could be observed in recent years. In this regard, an increase in the as yet relatively low utilisation rates of biobanks has been formulated as a goal. Higher utilisation rates can only be achieved if the perspectives of potential users of biobanks-particularly researchers not yet collaborating with biobanks-are adequately considered. To better understand their perspectives, a survey was conducted at ten different research institutions in Germany hosting a centralised biobank. The survey targeted potential users of biobank services, i.e. researchers working with biosamples. It addressed the general demand for biosamples, strategies for biosample acquisition/storage and reasons for/against collaborating with biobanks. In total, 354 researchers filled out the survey. Most interestingly, only a minority of researchers (12%) acquired their biosamples via biobanks. Of the respondents not collaborating with biobanks on sample acquisition, around half were not aware of the (services of the) respective local biobank. Those who actively decided against acquiring biosamples via a biobank provided different reasons. Most commonly, respondents stated that the biosamples required were not available, the costs were too high and information about the available biosamples was not readily accessible. Biobanks can draw many lessons from the results of the survey. Particularly, external communication and outreach should be improved. Additionally, biobanks might have to reassess whether their particular collection strategies are adequately aligned with local researchers' needs

A Novel System of Cytoskeletal Elements in the Human Pathogen Helicobacter pylori

Pathogenicity of the human pathogen Helicobacter pylori relies upon its capacity to adapt to a hostile environment and to escape from the host response. Therefore, cell shape, motility, and pH homeostasis of these bacteria are specifically adapted to the gastric mucus. We have found that the helical shape of H. pylori depends on coiled coil rich proteins (Ccrp), which form extended filamentous structures in vitro and in vivo, and are differentially required for the maintenance of cell morphology. We have developed an in vivo localization system for this pathogen. Consistent with a cytoskeleton-like structure, Ccrp proteins localized in a regular punctuate and static pattern within H. pylori cells. Ccrp genes show a high degree of sequence variation, which could be the reason for the morphological diversity between H. pylori strains. In contrast to other bacteria, the actin-like MreB protein is dispensable for viability in H. pylori, and does not affect cell shape, but cell length and chromosome segregation. In addition, mreB mutant cells displayed significantly reduced urease activity, and thus compromise a major pathogenicity factor of H. pylori. Our findings reveal that Ccrp proteins, but not MreB, affect cell morphology, while both cytoskeletal components affect the development of pathogenicity factors and/or cell cycle progression

Cortical thickness and resting-state cardiac function across the lifespan: a cross-sectional pooled mega analysis

Understanding the association between autonomic nervous system [ANS] function and brain morphology across the lifespan provides important insights into neurovisceral mechanisms underlying health and disease. Resting state ANS activity, indexed by measures of heart rate [HR] and its variability [HRV] has been associated with brain morphology, particularly cortical thickness [CT]. While findings have been mixed regarding the anatomical distribution and direction of the associations, these inconsistencies may be due to sex and age differences in HR/HRV and CT. Previous studies have been limited by small sample sizes, which impede the assessment of sex differences and aging effects on the association between ANS function and CT. To overcome these limitations, 20 groups worldwide contributed data collected under similar protocols of CT assessment and HR/HRV recording to be pooled in a mega-analysis (N = 1,218 (50.5% female), mean age 36.7 years (range: 12-87)). Findings suggest a decline in HRV as well as CT with increasing age. CT, particularly in the orbitofrontal cortex, explained additional variance in HRV, beyond the effects of aging. This pattern of results may suggest that the decline in HRV with increasing age is related to a decline in orbitofrontal CT. These effects were independent of sex and specific to HRV; with no significant association between CT and HR. Greater CT across the adult lifespan may be vital for the maintenance of healthy cardiac regulation via the ANS – or greater cardiac vagal activity as indirectly reflected in HRV may slow brain atrophy. Findings reveal an important association between cortical thickness and cardiac parasympathetic activity with implications for healthy aging and longevity that should be studied further in longitudinal research

Western blot analysis of CagY and CagT in KE and KE-59PCAT respectively.

<p>Western blot analysis of CagY and CagT in KE and KE-59PCAT respectively.</p

A) AGS cells co-incubated either with wild-type <i>H</i>. <i>pylori</i> cells (KE), <i>ccrp</i> deletion mutants as indicated or uninfected (AGS).

<p>Co-incubation was performed at MOI of 100 for 4 h. Cells were visualized by phase-contrast microscopy (BZ-9000E (KEYENCE) microscope) to assess AGS cell morphology. Scale bar, 20 μm. B) Quantification of the percentage of elongated cells from (A). All samples were examined in triplicate in at least three independent experiments. Data are presented as mean value of three independent experiments. For each strain between 1830 and 2800 cells were counted and evaluated. Exact percentage values are indicated above the bars. Asterisks indicate a significant difference between the <i>ccrp</i> mutants and wild-type <i>H</i>. <i>pylori</i> (the P value was <0.001, as determined by Student's t test). C) Bacterial adherence analysis in AGS cells infected with KE88-3887 or <i>ccrp</i> deletion mutants as indicated. AGS cells were infected with <i>H</i>. <i>pylori</i> for 3 h and 6 h, respectively. The number of cfu per cell was determined as described in experimental procedures and normalized to ml.</p

Quantification of CagT localization in immunostaining micrographs.

<p>Quantification of CagT localization in immunostaining micrographs.</p

A) Urease activity of strain KE88-3887 wild-type and <i>ccrp</i> mutants.

<p>Activity is expressed as percentage relative to wild-type in unsupplemented medium (set at 100%; no error bar). Results shown are the averages of six independent experiments; error bars denote standard deviations. Exact percentage values are indicated above the bars. Asterisks indicate a significant difference in urease activity between <i>ccrp</i> mutants and wild-type <i>H</i>. <i>pylori</i> (the P value was <0.001, as determined by Student's t test). B) Western blot using urease specific antiserum and strains as indicated above the lanes. Equal amounts of protein were loaded onto each lane. C) Urease activity of the wild-type strain and the <i>ccrp59</i> mutant in unsupplemented and nickel-supplemented media (as indicated in μM) shown as a percentage relative to wild-type in unsupplemented medium (set at 100%; no error bar). Results shown are the averages of six independent growth experiments; error bars denote standard deviations. Exact percentage values are indicated above the bars. Asterisks indicate a significant difference in urease activity between ccrp mutants and wild-type <i>H</i>. <i>pylori</i> (the P value was <0.001, as determined by Student's t test).</p

Strains, plasmids and primers used in this study.

<p>Strains, plasmids and primers used in this study.</p

A) Analysis of CagA expression and CagA tyrosine phosphorylation in the <i>ccrp59</i> mutant.

<p>Bacterial lysates from <i>H</i>. <i>pylori</i> wild type KE and <i>ccrp59</i> mutant KE-59PCAT were prepared respectively. Each sample that consisted of equivalent amounts of protein was subjected to immunoblotting assay using antiserum against CagA. AGS cells were not infected or infected with these strains at an MOI of 100 for 4h and subjected to immunoblotting analysis using specific antibody against phosphorylated CagA (Cag-P). B) Control experiment showing that CagA of the <i>ccrp59</i> mutant can be phosphorylated in vitro by mixing <i>H</i>. <i>pylori</i> lysates with AGS cell lysates.</p