Six shades of vascular smooth muscle cells illuminated by klf4 (krüppel-like factor 4)

Multiple layers of vascular smooth muscle cells (vSMCs) are present in blood vessels forming the media of the vessel wall. vSMCs provide a vessel wall structure, enabling it to contract and relax, thus modulating blood flow. They also play a crucial role in the development of vascular diseases, such as atherosclerosis and aortic aneurysm formation. vSMCs display a remarkable high degree of plasticity. At present, the number of different vSMC phenotypes has only partially been characterized. By mapping vSMC phenotypes in detail and identifying triggers for phenotype switching, the relevance of the different phenotypes in vascular disease may be identified. Up until recently, vSMCs were classified as either contractile or dedifferentiated (ie, synthetic). However, single-cell RNA sequencing studies revealed such dedifferentiated arterial vSMCs to be highly diverse. Currently, no consensus exist about the number of vSMC phenotypes. Therefore, we reviewed the data from relevant single-cell RNA sequencing studies, and classified a total of 6 vSMC phenotypes. The central dedifferentiated vSMC type that we classified is the mesenchymal-like phenotype. Mesenchymal-like vSMCs subsequently seem to differentiate into fibroblast-like, macrophage-like, osteogenic-like, and adipocyte-like vSMCs, which contribute differentially to vascular disease. This phenotype switching between vSMCs requires the transcription factor KLF4 (Krüppel-like factor 4). Here, we performed an integrated analysis of the data about the recently identified vSMC phenotypes, their associated gene expression profiles, and previous vSMC knowledge to better understand the role of vSMC phenotype transitions in vascular pathology

Six shades of vascular smooth muscle cells illuminated by klf4 (krüppel-like factor 4)

Multiple layers of vascular smooth muscle cells (vSMCs) are present in blood vessels forming the media of the vessel wall. vSMCs provide a vessel wall structure, enabling it to contract and relax, thus modulating blood flow. They also play a crucial role in the development of vascular diseases, such as atherosclerosis and aortic aneurysm formation. vSMCs display a remarkable high degree of plasticity. At present, the number of different vSMC phenotypes has only partially been characterized. By mapping vSMC phenotypes in detail and identifying triggers for phenotype switching, the relevance of the different phenotypes in vascular disease may be identified. Up until recently, vSMCs were classified as either contractile or dedifferentiated (ie, synthetic). However, single-cell RNA sequencing studies revealed such dedifferentiated arterial vSMCs to be highly diverse. Currently, no consensus exist about the number of vSMC phenotypes. Therefore, we reviewed the data from relevant single-cell RNA sequencing studies, and classified a total of 6 vSMC phenotypes. The central dedifferentiated vSMC type that we classified is the mesenchymal-like phenotype. Mesenchymal-like vSMCs subsequently seem to differentiate into fibroblast-like, macrophage-like, osteogenic-like, and adipocyte-like vSMCs, which contribute differentially to vascular disease. This phenotype switching between vSMCs requires the transcription factor KLF4 (Krüppel-like factor 4). Here, we performed an integrated analysis of the data about the recently identified vSMC phenotypes, their associated gene expression profiles, and previous vSMC knowledge to better understand the role of vSMC phenotype transitions in vascular pathology

Critical review of the use and scientific basis of forensic gait analysis

This review summarizes the scientific basis of forensic gait analysis and evaluates its use in the Netherlands, United Kingdom and Denmark, following recent critique on the admission of gait evidence in Canada. A useful forensic feature is (1) measurable, (2) consistent within and (3) different between individuals. Reviewing the academic literature, this article found that (1) forensic gait features can be quantified or observed from surveillance video, but research into accuracy, validity and reliability of these methods is needed; (2) gait is variable within individuals under differing and constant circumstances, with speed having major influence; (3) the discriminative strength of gait features needs more research, although clearly variation exists between individuals. Nevertheless, forensic gait analysis has contributed to several criminal trials in Europe in the past 15 years. The admission of gait evidence differs between courts. The methods are mainly observer-based: multiple gait analysts (independently) assess gait features on video footage of a perpetrator and suspect. Using gait feature databases, likelihood ratios of the hypotheses that the observed individuals have the same or another identity can be calculated. Automated gait recognition algorithms calculate a difference measure between video clips, which is compared with a threshold value derived from a video gait recognition database to indicate likelihood. However, only partly automated algorithms have been used in practice. We argue that the scientific basis of forensic gait analysis is limited. However, gait feature databases enable its use in court for supportive evidence with relatively low evidential value. The recommendations made in this review are (1) to expand knowledge on inter- and intra-subject gait variabilities, discriminative strength and interdependency of gait features, method accuracies, gait feature databases and likelihood ratio estimations; (2) to compare automated and observer-based gait recognition methods; to design (3) an international standard method with known validity, reliability and proficiency tests for analysts; (4) an international standard gait feature data collection method resulting in database(s); (5) (inter)national guidelines for the admission of gait evidence in court; and (6) to decrease the risk for cognitive and contextual bias in forensic gait analysis. This is expected to improve admission of gait evidence in court and judgment of its evidential value. Several ongoing research projects focus on parts of these recommendations

Endothelial dysfunction in Marfan syndrome mice is restored by resveratrol

Abstract Patients with Marfan syndrome (MFS) develop thoracic aortic aneurysms as the aorta presents excessive elastin breaks, fibrosis, and vascular smooth muscle cell (vSMC) death due to mutations in the FBN1 gene. Despite elaborate vSMC to aortic endothelial cell (EC) signaling, the contribution of ECs to the development of aortic pathology remains largely unresolved. The aim of this study is to investigate the EC properties in Fbn1C1041G/+ MFS mice. Using en face immunofluorescence confocal microscopy, we showed that EC alignment with blood flow was reduced, EC roundness was increased, individual EC surface area was larger, and EC junctional linearity was decreased in aortae of Fbn1C1041G/+ MFS mice. This modified EC phenotype was most prominent in the ascending aorta and occurred before aortic dilatation. To reverse EC morphology, we performed treatment with resveratrol. This restored EC blood flow alignment, junctional linearity, phospho-eNOS expression, and improved the structural integrity of the internal elastic lamina of Fbn1C1041G/+ mice. In conclusion, these experiments identify the involvement of ECs and underlying internal elastic lamina in MFS aortic pathology, which could act as potential target for future MFS pharmacotherapies

Endothelial dysfunction in Marfan syndrome mice is restored by resveratrol

Patients with Marfan syndrome (MFS) develop thoracic aortic aneurysms as the aorta presents excessive elastin breaks, fibrosis, and vascular smooth muscle cell (vSMC) death due to mutations in the FBN1 gene. Despite elaborate vSMC to aortic endothelial cell (EC) signaling, the contribution of ECs to the development of aortic pathology remains largely unresolved. The aim of this study is to investigate the EC properties in Fbn1C1041G/+ MFS mice. Using en face immunofluorescence confocal microscopy, we showed that EC alignment with blood flow was reduced, EC roundness was increased, individual EC surface area was larger, and EC junctional linearity was decreased in aortae of Fbn1C1041G/+ MFS mice. This modified EC phenotype was most prominent in the ascending aorta and occurred before aortic dilatation. To reverse EC morphology, we performed treatment with resveratrol. This restored EC blood flow alignment, junctional linearity, phospho-eNOS expression, and improved the structural integrity of the internal elastic lamina of Fbn1C1041G/+ mice. In conclusion, these experiments identify the involvement of ECs and underlying internal elastic lamina in MFS aortic pathology, which could act as potential target for future MFS pharmacotherapies

Improved epidermal barrier formation in human skin models by chitosan modulated dermal matrices

<div><p>Full thickness human skin models (FTMs) contain an epidermal and a dermal equivalent. The latter is composed of a collagen dermal matrix which harbours fibroblasts. Current epidermal barrier properties of FTMs do not fully resemble that of native human skin (NHS), which makes these human skin models less suitable for barrier related studies. To further enhance the resemblance of NHS for epidermal morphogenesis and barrier formation, we modulated the collagen dermal matrix with the biocompatible polymer chitosan. Herein, we report that these collagen-chitosan FTMs (CC-FTMs) possess a well-organized epidermis and maintain both the early and late differentiation programs as in FTMs. Distinctively, the epidermal cell activation is reduced in CC-FTMs to levels observed in NHS. Dermal-epidermal interactions are functional in both FTM types, based on the formation of the basement membrane. Evaluation of the barrier structure by the organization of the extracellular lipid matrix of the stratum corneum revealed an elongated repeat distance of the long periodicity phase. The ceramide composition exhibited a higher resemblance of the NHS, based on the carbon chain-length distribution and subclass profile. The inside-out barrier functionality indicated by the transepidermal water loss is significantly improved in the CC-FTMs. The expression of epidermal barrier lipid processing enzymes is marginally affected, although more restricted to a single granular layer. The novel CC-FTM resembles the NHS more closely, which makes them a promising tool for epidermal barrier related studies.</p></div

Ceramide subclass profile in FTMs and CC-FTMs.

<p>Box and whisker plot of CER subclasses with indicated benchmark values of NHS. Significant differences in relative abundance are indicated, which are the reduction in NS and AS and an increase of NdS, NP and NH in SC of CC-FTMs compared to FTMs. Whiskers indicate the 95% confidence interval. Data is obtained from four independent experiments.</p

Expression of lipid processing enzymes in NHS, FTM and CC-FTM.

<p>Tissue sections of NHS, FTMs and CC-FTMs are analysed by immunofluorescence for the expression of lipid processing enzymes GBA, aSMase, CER-S3 and ELOVL1. Representative images show the localization (red) of GBA, aSMase and CER-S3 in the stratum granulosum, while the expression of ELOVL1 is more diffuse throughout the epidermis. Nuclei are stained blue using DAPI, the yellow dotted line indicates the dermal-epidermal junction. Scale bar represents 100μm.</p

Transepidermal water loss of NHS, FTMs and CC-FTMs.

<p>TEWL was measured over 900 second time period and plotted as mean values +SEM. Data is obtained from three independent experiments and three NHS samples.</p