Twelve tips for creating a longitudinal quality improvement and safety education for early health professions students.

Despite the numerous calls for integrating quality improvement and patient safety (QIPS) curricula into health professions education, there are limited examples of effective implementation for early learners. Typically, pre-clinical QIPS experiences involve lectures or lessons that are disconnected from the practice of medicine. Consequently, students often prioritize other content they consider more important. As a result, they may enter clinical settings without essential QIPS skills and struggle to incorporate these concepts into their early professional identity formation. In this paper, we present twelve tips aimed at assisting educators in developing QIPS education early in the curricula of health professions students. These tips address various key issues, including aligning incentives, providing longitudinal experiences, incorporating real-world care outcomes, optimizing learning environments, communicating successes, and continually enhancing education and care delivery processes.

Life cycle assessment of advanced grade PLA product with novel end-of-life treatment through depolymerization.

Using biobased plastics has the potential to avoid fossil resource depletion and fossil CO2 emissions. Polylactic acid (PLA) is a fast-growing bio-based plastic made from fermented sugars. Nowadays, PLA is used to replace fossil-based polymers in healthcare and single-use applications, such as for packaging applications. However, PLA offers a much broader application range with the targeted use of a combination of its stereoisomers; PL(L)A and PL(D)A. A variety of these advanced grades of PLA can be used for multiple purposes in durable consumer products such as furniture. Recycling complex, mixed material and advanced grades of PLA is currently limited, as mechanical recycling has limitations in recycling mixed PLA grades. Using a depolymerization technology, products of such advanced grades of PLA can be recycled to form high-quality recycled PLA. A cradle-to-grave life cycle assessment study was executed to evaluate the sustainability of high-end durable product (a rug) with mixtures of PLA grade and the novel depolymerization technology. The findings of the study showed a 70 % reduction in CO2-eq. emissions compared to a conventionally designed rug. However, an increase is indicated in the following environmental impact categories: land use, eutrophication, and environmental toxicity. Sensitivity analyses for collection rates showcased that design for collection and recycling are key to obtaining a more sustainable biobased products. Additionally, scenario analysis supported depolymerization for PLA as recycling technology with low CO2-eq. emissions. Based on the results of the LCA and additional scenario analysis, the use of PLA is encouraged to be used in more durable and lasting products, such as furniture, from an environmental perspective, provided that the products are designed for collection and high-quality recycling to ensure material circularity.

Human In Vitro Skin Models for Wound Healing and Wound Healing Disorders.

Skin wound healing is essential to health and survival. Consequently, high amounts of research effort have been put into investigating the cellular and molecular components involved in the wound healing process. The use of animal experiments has contributed greatly to the knowledge of wound healing, skin diseases, and the exploration of treatment options. However, in addition to ethical concerns, anatomical and physiological inter-species differences often influence the translatability of animal-based studies. Human in vitro skin models, which include essential cellular and structural components for wound healing analyses, would improve the translatability of results and reduce animal experiments during the preclinical evaluation of novel therapy approaches. In this review, we summarize in vitro approaches, which are used to study wound healing as well as wound healing-pathologies such as chronic wounds, keloids, and hypertrophic scars in a human setting.

Modelling the Complexity of Human Skin In Vitro.

The skin serves as an important barrier protecting the body from physical, chemical and pathogenic hazards as well as regulating the bi-directional transport of water, ions and nutrients. In order to improve the knowledge on skin structure and function as well as on skin diseases, animal experiments are often employed, but anatomical as well as physiological interspecies differences may result in poor translatability of animal-based data to the clinical situation. In vitro models, such as human reconstructed epidermis or full skin equivalents, are valuable alternatives to animal experiments. Enormous advances have been achieved in establishing skin models of increasing complexity in the past. In this review, human skin structures are described as well as the fast evolving technologies developed to reconstruct the complexity of human skin structures in vitro.

Rosiglitazone Reverses Inflammation in Epididymal White Adipose Tissue in Hormone-Sensitive Lipase-Knockout Mice.

Hormone-sensitive lipase (HSL) plays a crucial role in intracellular lipolysis, and loss of HSL leads to diacylglycerol (DAG) accumulation, reduced FA mobilization, and impaired PPARγ signaling. Hsl knockout mice exhibit adipose tissue inflammation, but the underlying mechanisms are still not clear. Here, we investigated if and to what extent HSL loss contributes to endoplasmic reticulum (ER) stress and adipose tissue inflammation in Hsl knockout mice. Furthermore, we were interested in how impaired PPARγ signaling affects the development of inflammation in epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT) of Hsl knockout mice and if DAG and ceramide accumulation contribute to adipose tissue inflammation and ER stress. Ultrastructural analysis showed a markedly dilated ER in both eWAT and iWAT upon loss of HSL. In addition, Hsl knockout mice exhibited macrophage infiltration and increased F4/80 mRNA expression, a marker of macrophage activation, in eWAT, but not in iWAT. We show that treatment with rosiglitazone, a PPARγ agonist, attenuated macrophage infiltration and ameliorated inflammation of eWAT, but expression of ER stress markers remained unchanged, as did DAG and ceramide levels in eWAT. Taken together, we show that HSL loss promoted ER stress in both eWAT and iWAT of Hsl knockout mice, but inflammation and macrophage infiltration occurred mainly in eWAT. Also, PPARγ activation reversed inflammation but not ER stress and DAG accumulation. These data indicate that neither reduction of DAG levels nor ER stress contribute to the reversal of eWAT inflammation in Hsl knockout mice.

Process Technologies of Cyanobacteria.

Although the handling and exploitation of cyanobacteria is associated with some challenges, these phototrophic bacteria offer great opportunities for innovative biotechnological processes. This chapter covers versatile aspects of working with cyanobacteria, starting with up-to-date in silico and in vitro screening methods for bioactive substances. Subsequently, common conservation techniques and vitality/viability estimation methods are compared and supplemented by own data regarding the non-invasive vitality evaluation via pulse amplitude modulated fluorometry. Moreover, novel findings about the influence the state of the pre-cultures have on main cultures are presented. The following sub-chapters deal with different photobioreactor-designs, with special regard to biofilm photobioreactors, as well as with heterotrophic and mixotrophic cultivation modes. The latter topic provides information from literature on successfully enhanced cyanobacterial production processes, augmented by own data.

Autologous Fat Grafting in Reconstructive Breast Surgery: Clinically Relevant Factors Affecting the Graft Take.

BACKGROUND: Autologous fat grafting is an effective tool for soft tissue augmentation in reconstructive breast surgery. Despite the major advantages of this minimally invasive approach, the unpredictability of graft survival presents challenges. OBJECTIVES: No clear consensus on the optimal technique has yet been published and well-defined prospective studies investigating impairing factors are lacking. This aim of this study was to generate valuable fundamental data. METHODS: Ten female patients undergoing elective autologous fat grafting after nipple-sparing mastectomy were enrolled. Punch biopsies and lipoaspirates were collected from the harvest site for histologic, gene expression, and scanning electron microscopic analysis. Noninvasive Lipometer measurements determining the subcutaneous adipose tissue thickness at the graft site were used to calculate the respective take rate. Patient- and surgery-related data were acquired and correlated with the take rate. RESULTS: A statistically relevant correlation between the take rate and the existing mean subcutaneous adipose tissue thickness at the grafted breast prior to surgery was observed. An approximate correlation was identified regarding the number of previous grafting sessions, body weight, and BMI. No statistically significant correlation was demonstrated for age, harvest site, or the mean adipocyte size. A lower level of cell damage was observed in scanning electron microscopic samples of washed lipoaspirates; and a strong indirect correlation with the expression of the adipocyte markers FABP4 and PLIN1 was apparent. CONCLUSIONS: Factors correlating to the take rate were identified. Future studies investigating the clinical relevance of each impairing factor are essential to contribute to the optimization of this valuable method.

Tracking the nature and trajectory of social support in Facebook mutual aid groups during the COVID-19 pandemic.

At the onset of the COVID-19 pandemic, thousands of mutual aid groups were established on social media and operated as platforms through which people could offer or request social support. Considering the importance of Facebook mutual aid groups during the early stages of the COVID-19 pandemic in the United Kingdom but also the lack of empirical research regarding the trajectories and types of social support rendered available through the groups, our aims in this paper are threefold; first, to examine the trajectory of social support-related activity during the period between March-December 2020; second, to compare offers and requests of support during the peaks of the first and second waves; third to provide a rich analysis of the types of social support that were offered or requested through the online mutual aid groups. Quantitative findings suggest that online social support activity declined soon after the peak of the first pandemic wave and, at least in Facebook mutual aid groups, did not reach the levels observed during the first wave. Also, the number of offers of support during the first wave was higher compared to offers during the second wave, and similar was the case for requests for support. Additionally, offers for support were higher compared to requests for support during both the first and second waves. Finally, qualitative analysis showed that people used the Facebook mutual aid groups to offer and request various types of practical, emotional, and informational support. Limitations as well as implications of our study are considered.

Correlation between ranges of leg walking angles and passive rest angles among leg types in stick insects.

Because of scaling issues, passive muscle and joint forces become increasingly important as limb size decreases.1-3 In some small limbs, passive forces can drive swing in locomotion,4,5 and antagonist passive torques help control limb swing velocity.6 In stance, minimizing antagonist muscle and joint passive forces could save energy. These considerations predict that, for small limbs, evolution would result in the angle range over which passive forces are too small to cause limb movement (called "resting-state range" in prior insect work4 and "area of neutral equilibrium" in physics and engineering) correlating with the limb's typical working range, usually that in locomotion. We measured the most protracted and retracted thorax-femur (ThF) angles of the pro- (front), meso- (middle), and metathoracic (hind) leg during stick insect (Carausius morosus) walks. This ThF working range differed in the three leg types, being more posterior in more posterior legs. In other experiments, we manually protracted or retracted the denervated front, middle, and hind legs. Upon release, passive forces moved the leg in the opposite direction (retraction or protraction) until it reached the most protracted or most retracted edge of the ThF resting-state range. The ThF resting-state angle ranges correlated with the leg-type working range, being more posterior in more posterior legs. The most protracted ThF walking angles were more retracted than the post-protraction ThF angles, and the most retracted ThF walking angles were similar to the post-retraction ThF angles. These correlations of ThF working- and resting-state ranges could simplify motor control and save energy. These data also provide an example of evolution altering behavior by changing passive muscle and joint properties.7.

Passively immobilized cyanobacteria Nostoc species BB 92.2 in a moving bed photobioreactor (MBPBR): Design, cultivation, and characterization.

The cyanobacterium Nostoc sp. BB 92.3. had shown antibacterial activity. A cultivation as biofilm, a self-forming matrix of cells and extracellular polymeric substances, increased the antibacterial effect. A new photobioreactor system was developed that allows a surface-associated cultivation of Nostoc sp. as biofilm. High-density polyethylene carriers operated as a moving bed were selected as surface for biomass immobilization. This system, well established in heterotrophic wastewater treatment, was for the first time used for phototrophic biofilms. The aim was a cultivation on a large scale without inhibiting growth while maximizing immobilization. Cultivation in a small photobioreactor (1.5 L) with different volumetric filling degrees of carriers (13.4%-53.8%) in a batch process achieved immobilization rates of 70%-85% and growth was similar to a no-carrier-control. In a larger photobioreactor (65 L) essentially all of the biomass was immobilized on the carriers and the space-time yield of biomass (0.018 gcell dry weight L-1 day- ​​​​​​​1 ) was competitive compared to phototrophic biofilm cultivations from literature. The use of carriers increased the gas exchange in the reactor by a factor of 2.5-3 but doubled the mixing time. Enriched gassing with carbon dioxide resulted in a short-term increase in growth rate, but unexpectedly it also adversely changed the growth morphology.

More Than a COVID-19 Response: Sustaining Mutual Aid Groups During and Beyond the Pandemic.

Mutual aid groups have been an indispensable part of the public response to the COVID-19 pandemic. They have provided many forms of support, in particular grocery shopping which has enabled people to self-isolate if required. While community solidarity during emergencies and disasters is common, previous studies have shown that such solidarity behaviors tend to decline over time, even when needs remain high. In this study, we address how mutual aid groups can be sustained over time in the context of the COVID-19 pandemic. We conducted 32 interviews with organizers of COVID-19 mutual aid and community support groups in the United Kingdom between September 2020 and January 2021. Based on a reflexive thematic analysis, we identified several community and group level experiences and strategies that were related to sustained participation in COVID-19 mutual aid groups. Meeting community needs over time with localized action and resources and building trust and community-based alliances were foundational elements in the COVID-19 mutual aid groups. Group processes strategies, such as a culture of care and support and regular group meetings, were used to help to sustain involvement. Some experiences resulting from participation in COVID-19 mutual aid groups were also related to sustained participation, including positive emotions (e.g., joy, pride), well-being and sense of efficacy, and an increasing sense of local community belonging and cohesion. Based on these findings, we propose four practical recommendations for sustaining mutual aid groups to assist public engagement with protective behaviors in the COVID-19 pandemic and beyond. We recommend providing practical and financial support to COVID-19 mutual aid groups; to mobilize the knowledge and the experiences acquired by COVID-19 mutual aid groups for developing programs and interventions for addressing the medium and long-term impacts of COVID-19; to prioritize community-level interventions; and to recognize the role of group processes as these have the potential to lead to long-term community responses. These approaches will be key for ensuring that communities effectively recover from the COVID-19 pandemic.

New procedure for separation and analysis of the main components of cyanobacterial EPS.

Phototrophic biofilms produce a matrix of extracellular polymeric substances (EPS), which holds the cells together and functions inter alia as nutrient storage and protection layer. EPS mainly consist of water, polysaccharides, proteins, lipids and nucleic acids as well as lysis and hydrolysis products which makes the composition very complex. Thus, rough simplifications are used and commonly one or at most two components of the EPS are examined. In this work a new procedure for separation and analysis of EPS in the main components (i) polysaccharides, (ii) proteins and (iii) lipids is presented with recovery rates of nearly 100 %. The method was established with synthetic EPS, which based on the composition of real EPS described in literature. Afterwards, the method was transferred to real EPS samples allowing a deeper insight in the composition of EPS from only one sample. The composition of EPS-extracts from Nostoc spec, cultivated under heterotrophic and mixotrophic batch and fed-batch conditions, was analysed during a cultivation period of 14 days. It was observed that mixotrophic cultivation led to higher amounts of carbohydrates, lipids and proteins than heterotrophic cultivation respectively, regardless of batch or fed-batch culture. While the amount of proteins in the EPS increased during the cultivation period, carbohydrates and lipids were dominant in the beginning and decreased afterwards.

Cell-free in vitro reduction of carboxylates to aldehydes: With crude enzyme preparations to a key pharmaceutical building block.

The scarcity of practical methods for aldehyde synthesis in chemistry necessitates the development of mild, selective procedures. Carboxylic acid reductases catalyze aldehyde formation from stable carboxylic acid precursors in an aqueous solution. Carboxylic acid reductases were employed to catalyze aldehyde formation in a cell-free system with activation energy and reducing equivalents provided through auxiliary proteins for ATP and NADPH recycling. In situ product removal was used to suppress over-reduction due to background enzyme activities, and an N-protected 4-formyl-piperidine pharma synthon was prepared in 61% isolated yield. This is the first report of preparative aldehyde synthesis with carboxylic acid reductases employing crude, commercially available enzyme preparations.

Novel method enabling a rapid vitality determination of cyanobacteria.

Cyanobacteria represent a large group of bacteria with underestimated scientific potential. Recent studies indicate them as a great reservoir of secondary metabolites with antifungal, antiviral or antibacterial activity. However, common, well established research techniques cannot be easily adapted to these organisms. Slow growth rates and irregular cell aggregates constitute challenges for researchers dealing with cyanobacteria. In this work, we present an innovative new method enabling a quick, easy and economical vitality determination of cyanobacterial strains, as, e.g. required for the finding of optimal cryopreservation conditions. We were able to measure the vitality of previously cryopreserved and defrosted Trichocoleus sociatus samples within 45 min by means of their O2-production. For each run, a cell wet mass of only 0.5 g was required. By application of this method, we could find DMSO (5% v/v) and glycerin (15% v/v) to be the most promising cryoprotectants for the conservation of T. sociatus cells. DMSO and glycerin guaranteed a vitality rate of 80-90% and 60-70% after up to four weeks of cryopreservation, compared to fresh cell material.

Premature Activation of Immune Transcription Programs in Autoimmune-Predisposed Mouse Embryonic Stem Cells and Blastocysts.

Autoimmune diabetes is a complex multifactorial disease with genetic and environmental factors playing pivotal roles. While many genes associated with the risk of diabetes have been identified to date, the mechanisms by which external triggers contribute to the genetic predisposition remain unclear. Here, we derived embryonic stem (ES) cell lines from diabetes-prone non-obese diabetic (NOD) and healthy C57BL/6 (B6) mice. While overall pluripotency markers were indistinguishable between newly derived NOD and B6 ES cells, we discovered several differentially expressed genes that normally are not expressed in ES cells. Several genes that reside in previously identified insulin-dependent diabetics (Idd) genomic regions were up-regulated in NOD ES cells. Gene set enrichment analysis showed that different groups of genes associated with immune functions are differentially expressed in NOD. Transcriptomic analysis of NOD blastocysts validated several differentially overexpressed Idd genes compared to B6. Genome-wide mapping of active histone modifications using ChIP-Seq supports active expression as the promoters and enhancers of activated genes are also marked by active histone modifications. We have also found that NOD ES cells secrete more inflammatory cytokines. Our data suggest that the known genetic predisposition of NOD to autoimmune diabetes leads to epigenetic instability of several Idd regions.

Co-factor demand and regeneration in the enzymatic one-step reduction of carboxylates to aldehydes in cell-free systems.

Addressing the challenges associated with the development of in vitro biocatalytic carboxylate reductions for potential applications, important aspects of the co-factor regeneration systems and strategies for minimizing over-reduction were investigated. The ATP recycling can be performed with similarly high efficiency exploiting the polyphosphate source by combining Meiothermus ruber polyphosphate kinase and adenylate kinase or with Sinorhizobium meliloti polyphosphate kinase instead of the latter. Carboxylate reductions with the enzyme candidates used in this work allow operating at co-factor concentrations of adenosine 5'-triphosphate and ß-nicotinamide adenine dinucleotide 2'-phosphate of 100 µM and, thereby, reducing the amounts of alcohols formed by side activities in the enzyme preparations. This study confirmed the expected benefits of carboxylic acid reductases in chemoselectively reducing the carboxylates to the corresponding aldehydes while leaving reductively-sensitive nitro, ester and cyano groups intact.

In situ analysis of male meiosis in C. elegans.

We describe a routine method to locate cells of appropriate meiotic stages in the gonad of Caenorhabditis elegans males prior to 3D reconstruction of meiotic spindles by electron tomography. For this, serial semi-thick (300nm) sections of whole worms are pre-screened and recorded at low magnification by transmission electron microscopy. Cells of interest are identified in aligned image stacks showing the entire proximal region of male gonads at low magnification. Tilt series of selected cells are then recorded at higher magnification to reconstruct meiotic spindles of selected cells in 3D. Our approach allows a routine staging of spermatocytes without the use of anesthetics or the application of physical immobilization of worms. We also describe a modification of a previously published protocol (Muller-Reichert, Srayko, Hyman, O'Toole, & McDonald, 2007) by using polyvinylpyrrolidone (PVP) instead of bovine serum albumin (BSA) as a "filler" for specimen loading in high-pressure freezing.

Enzymatic One-Step Reduction of Carboxylates to Aldehydes with Cell-Free Regeneration of ATP and NADPH.

The direct generation of aldehydes from carboxylic acids is often a challenging synthetic task but undoubtedly attractive in view of abundant supply of such feedstocks from nature. Though long known, biocatalytic carboxylate reductions are at an early stage of development, presumably because of their co-factor requirement. To establish an alternative to whole-cell-based carboxylate reductions which are limited by side reactions, we developed an in vitro multi-enzyme system that allows for quantitative reductions of various carboxylic acids with full recycling of all cofactors and prevention of undesired over-reductions. Regeneration of adenosine 5'-triphosphate is achieved through the simultaneous action of polyphosphate kinases from Meiothermus ruber and Sinorhizobium meliloti and ß-nicotinamide adenine dinucleotide 2'-phosphate is reduced by a glucose dehydrogenase. Under these conditions and in the presence of the carboxylate reductases from Neurospora crassa or Nocardia iowensis, various aromatic, heterocyclic and aliphatic carboxylic acids were quantitatively reduced to the respective aldehydes.

Atorvastatin, but not pravastatin, inhibits cardiac Akt/mTOR signaling and disturbs mitochondrial ultrastructure in cardiac myocytes.

Statins, which reduce LDL-cholesterol by inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, are among the most widely prescribed drugs. Skeletal myopathy is a known statin-induced adverse effect associated with mitochondrial changes. We hypothesized that similar effects would occur in cardiac myocytes in a lipophilicity-dependent manner between 2 common statins: atorvastatin (lipophilic) and pravastatin (hydrophilic). Neonatal cardiac ventricular myocytes were treated with atorvastatin and pravastatin for 48 h. Both statins induced endoplasmic reticular (ER) stress, but only atorvastatin inhibited ERK1/2T202/Y204, AktSer473, and mammalian target of rapamycin signaling; reduced protein abundance of caveolin-1, dystrophin, epidermal growth factor receptor, and insulin receptor-ß; decreased Ras homolog gene family member A activation; and induced apoptosis. In cardiomyocyte-equivalent HL-1 cells, atorvastatin, but not pravastatin, reduced mitochondrial oxygen consumption. When male mice underwent atorvastatin and pravastatin administration per os for up to 7 mo, only long-term atorvastatin, but not pravastatin, induced elevated serum creatine kinase; swollen, misaligned, size-variable, and disconnected cardiac mitochondria; alteration of ER structure; repression of mitochondria- and endoplasmic reticulum-related genes; and a 21% increase in mortality in cardiac-specific vinculin-knockout mice during the first 2 months of administration. To our knowledge, we are the first to demonstrate in vivo that long-term atorvastatin administration alters cardiac ultrastructure, a finding with important clinical implications.-Godoy, J. C., Niesman, I. R., Busija, A. R., Kassan, A., Schilling, J. M., Schwarz, A., Alvarez, E. A., Dalton, N. D., Drummond, J. C., Roth, D. M., Kararigas, G., Patel, H. H., Zemljic-Harpf, A. E. Atorvastatin, but not pravastatin, inhibits cardiac Akt/mTOR signaling and disturbs mitochondrial ultrastructure in cardiac myocytes.

Revisiting Centrioles in Nematodes-Historic Findings and Current Topics.

Theodor Boveri is considered as the "father" of centrosome biology. Boveri's fundamental findings have laid the groundwork for decades of research on centrosomes. Here, we briefly review his early work on centrosomes and his first description of the centriole. Mainly focusing on centriole structure, duplication, and centriole assembly factors in C. elegans, we will highlight the role of this model in studying germ line centrosomes in nematodes. Last but not least, we will point to future directions of the C. elegans centrosome field.