Amyloid Beta Influences Vascular Smooth Muscle Contractility and Mechanoadaptation.

Amyloid beta accumulation in neuronal and cerebrovascular tissue is a key precursor to development of Alzheimer's disease and can result in neurodegeneration. While its persistence in Alzheimer's cases is well-studied, amyloid beta's direct effect on vascular function is unclear. Here, we measured the effect of amyloid beta treatment on vascular smooth muscle cell functional contractility and modeled the mechanoadaptive growth and remodeling response to these functional perturbations. We found that the amyloid beta 1-42 isoform induced a reduction in vascular smooth muscle cell mechanical output and reduced response to vasocontractile cues. These data were used to develop a thin-walled constrained mixture arterial model that suggests vessel growth, and remodeling in response to amyloid betamediated alteration of smooth muscle function leads to decreased ability of cerebrovascular vessels to vasodilate. These findings provide a possible explanation for the vascular injury and malfunction often associated with the development of neurodegeneration in Alzheimer's disease.

Green tea extract enhances retinal ganglion cell survival and axonal regeneration in rats with optic nerve injury.

Current clinical treatments have not yet effectively cured progressive retinal ganglion cell (RGC) death and axonal degeneration after optic nerve (ON) injury. We previously demonstrated green tea extract (GTE) can reduce RGC death in rats after ischemic injury. Here, we aim to determine the prophylactic and therapeutic effects and mechanisms of GTE on RGC survival and axonal regeneration in rats with ON injury. GTE (275 or 550 mg/kg) was administered intragastrically for 7 d before or 14 d post-ON crush surgery in adult Fischer 344 rats. Rats with pre- or post-operative treatment of 275 mg/kg GTE showed significantly higher numbers of RGCs and regenerated axons post-ON injury with improved pupillary light reflex as compared to saline-treated rats. Akt and Erk p42/44 activation was higher in the retina of rats given 275 mg/kg GTE pre-surgery, whereas Stat3 activation was higher in those with 275 mg/kg GTE post-operation. Less activated microglia were observed in rats with pre-treatment of 275 or 550 mg/kg GTE. RNA sequencing analysis identified the downregulation of inflammation, apoptosis, and microglia activation genes in the retina of rats with pre- or post-treatment with 275 mg/kg GTE as compared to the saline-treated rats. In summary, this study revealed the prophylactic and therapeutic treatment effects of GTE on RGC survival and axonal regeneration in rats with ON injury, indicating a potential alternative treatment for traumatic optic neuropathy.

Efficient Malaria Parasite Detection From Diverse Images of Thick Blood Smears for Cross-Regional Model Accuracy.

Goal: The purpose of this work is to improve malaria diagnosis efficiency by integrating smartphones with microscopes. This integration involves image acquisition and algorithmic detection of malaria parasites in various thick blood smear (TBS) datasets sourced from different global regions, including low-quality images from Sub-Saharan Africa. Methods: This approach combines image segmentation and a convolutional neural network (CNN) to distinguish between white blood cells, artifacts, and malaria parasites. A portable system integrates a microscope with a graphical user interface to facilitate rapid malaria detection from smartphone images. We trained the CNN model using open-source data from the Chittagong Medical College Hospital, Bangladesh. Results: The validation process, using microscopic TBS from both the training dataset and an additional dataset from Sub-Saharan Africa, demonstrated that the proposed model achieved an accuracy of 97.74% ± 0.05% and an F1-score of 97.75% ± 0.04%. Remarkably, our proposed model with AlexNet surpasses the reported literature performance of 96.32%. Conclusions: This algorithm shows promise in aiding malaria-stricken regions, especially those with limited resources.

Microfluidic Genipin Deposition Technique for Extended Culture of Micropatterned Vascular Muscular Thin Films.

The chronic nature of vascular disease progression requires the development of experimental techniques that simulate physiologic and pathologic vascular behaviors on disease-relevant time scales. Previously, microcontact printing has been used to fabricate two-dimensional functional arterial mimics through patterning of extracellular matrix protein as guidance cues for tissue organization. Vascular muscular thin films utilized these mimics to assess functional contractility. However, the microcontact printing fabrication technique used typically incorporates hydrophobic PDMS substrates. As the tissue turns over the underlying extracellular matrix, new proteins must undergo a conformational change or denaturing in order to expose hydrophobic amino acid residues to the hydrophobic PDMS surfaces for attachment, resulting in altered matrix protein bioactivity, delamination, and death of the tissues. Here, we present a microfluidic deposition technique for patterning of the crosslinker compound genipin. Genipin serves as an intermediary between patterned tissues and PDMS substrates, allowing cells to deposit newly-synthesized extracellular matrix protein onto a more hydrophilic surface and remain attached to the PDMS substrates. We also show that extracellular matrix proteins can be patterned directly onto deposited genipin, allowing dictation of engineered tissue structure. Tissues fabricated with this technique show high fidelity in both structural alignment and contractile function of vascular smooth muscle tissue in a vascular muscular thin film model. This technique can be extended using other cell types and provides the framework for future study of chronic tissue- and organ-level functionality.

Vascular smooth muscle cell functional contractility depends on extracellular mechanical properties.

Vascular smooth muscle cells' primary function is to maintain vascular homeostasis through active contraction and relaxation. In diseases such as hypertension and atherosclerosis, this function is inhibited concurrent to changes in the mechanical environment surrounding vascular smooth muscle cells. It is well established that cell function and extracellular mechanics are interconnected; variations in substrate modulus affect cell migration, proliferation, and differentiation. To date, it is unknown how the evolving extracellular mechanical environment of vascular smooth muscle cells affects their contractile function. Here, we have built upon previous vascular muscular thin film technology to develop a variable-modulus vascular muscular thin film that measures vascular tissue functional contractility on substrates with a range of pathological and physiological moduli. Using this modified vascular muscular thin film, we found that vascular smooth muscle cells generated greater stress on substrates with higher moduli compared to substrates with lower moduli. We then measured protein markers typically thought to indicate a contractile phenotype in vascular smooth muscle cells and found that phenotype is unaffected by substrate modulus. These data suggest that mechanical properties of vascular smooth muscle cells' extracellular environment directly influence their functional behavior and do so without inducing phenotype switching.

Smooth muscle phenotype switching in blast traumatic brain injury-induced cerebral vasospasm.

Due to increased survival rates among soldiers exposed to explosive blasts, blast-induced traumatic brain injury (bTBI) has become much more prevalent in recent years. Cerebral vasospasm (CVS) is a common manifestation of brain injury whose incidence is significantly increased in bTBI. CVS is characterized by initial vascular smooth muscle cell (VSMC) hypercontractility, followed by prolonged vessel remodeling and lumen occlusion, and is traditionally associated with subarachnoid hemorrhage (SAH), but recent results suggest that mechanical injury during bTBI can cause mechanotransduced VSMC hypercontractility and phenotype switching necessary for CVS development, even in the absence of SAH. Here, we review the mechanisms by which mechanical stimulation and SAH can synergistically drive CVS progression, complicating treatment options in bTBI patients.

Long-term vascular contractility assay using genipin-modified muscular thin films.

Vascular disease is a leading cause of death globally and typically manifests chronically due to long-term maladaptive arterial growth and remodeling. To date, there is no in vitro technique for studying vascular function over relevant disease time courses that both mimics in vivo-like tissue structure and provides a simple readout of tissue stress. We aimed to extend tissue viability in our muscular thin film contractility assay by modifying the polydimethylsiloxane (PDMS) substrate with micropatterned genipin, allowing extracellular matrix turnover without cell loss. To achieve this, we developed a microfluidic delivery system to pattern genipin and extracellular matrix proteins on PDMS prior to cell seeding. Tissues constructed using this method showed improved viability and maintenance of in vivo-like lamellar structure. Functional contractility of tissues fabricated on genipin-modified substrates remained consistent throughout two weeks in culture. These results suggest that muscular thin films with genipin-modified PDMS substrates are a viable method for conducting functional studies of arterial growth and remodeling in vascular diseases.

Smooth muscle architecture within cell-dense vascular tissues influences functional contractility.

The role of vascular smooth muscle architecture in the function of healthy and dysfunctional vessels is poorly understood. We aimed at determining the relationship between vascular smooth muscle architecture and contractile output using engineered vascular tissues. We utilized microcontact printing and a microfluidic cell seeding technique to provide three different initial seeding conditions, with the aim of influencing the cellular architecture within the tissue. Cells seeded in each condition formed confluent and aligned tissues but within the tissues, the cellular architecture varied. Tissues with a more elongated cellular architecture had significantly elevated basal stress and produced more contractile stress in response to endothelin-1 stimulation. We also found a correlation between the contractile phenotype marker expression and the cellular architecture, contrary to our previous findings in non-confluent tissues. Taken with previous results, these data suggest that within cell-dense vascular tissues, smooth muscle contractility is strongly influenced by cell and tissue architectures.

Application of a digital head-posture measuring system in children.

PURPOSE: To report the repeatability of a digital head-posture measuring system when used to record anomalous head postures in children. DESIGN: Prospective study and clinical laboratory investigation. METHODS: Using a digital head-posture measuring system, we measured 36 different anomalous head postures in 27 children with infantile nystagmus syndrome. Repeatability values and 95% limits of repeatability of measurements were generated for anomalous head postures. RESULTS: Among the 27 children, 3 had 2 head postures (right and left head turns) in 2 different directions; 6 had 2-dimensional head postures that were considered 2 different head postures; and 18 had a 1-dimensional head posture. There were 5 chin-up or chin-down postures, 23 head-turn postures, and 8 head-tilt postures in a total of 36 anomalous head postures. The repeatability value for all anomalous head postures was less than 10 degrees. Ninety-five percent limits of repeatability yielded ranges of less than 10 degrees for all anomalous head postures. CONCLUSIONS: The digital head-posture measuring system is a valid and reliable device for measuring 3-dimensional head postures in children with nystagmus.

Collagen-agarose co-gels as a model for collagen-matrix interaction in soft tissues subjected to indentation.

The mechanical properties of soft tissues depend on the collagen fiber network and the surrounding non-fibrillar matrix. The mechanical role of non-fibrillar material remains poorly understood. Our recent study (Lake and Barocas, Ann Biomed Eng 2011) introduced collagen-agarose co-gels as a simple experimental model system to evaluate the mechanical contribution of non-fibrillar matrix, and evaluated co-gel properties in uniaxial tension. In this study, we utilized similar co-gels to examine collagen-matrix interaction in tissues subjected to incremental stress-relaxation indentation tests. Mechanical testing was performed using two orthogonal custom test devices, and polarized light imaging was used to quantify 3D collagen fiber kinematics under load. The addition of agarose led to concentration-dependent changes in the time-dependent mechanical response and magnitude/spread of collagen fiber reorganization of tissue analogs. Specifically, peak/relaxed loads increased, and relaxation rate decreased, with increasing agarose concentration. In addition, increasing agarose content led to larger magnitude changes in orientation direction and alignment strength that were more localized near the indenter. Results suggest that non-fibrillar material significantly contributes to the behavior of co-gels in indentation, likely by reducing permeability and resisting volume change, thereby providing insight into the properties of artificial and native tissues subjected to non-tensile loading.

Development and validation of a digital head posture measuring system.

PURPOSE: To report the accuracy and repeatability of a new digital head posture measuring system. DESIGN: Prospective study, clinical laboratory investigation. METHODS: The digital head posture measuring system consists of a head-mounted motion tracker, a standard personal computer, and customized software to sample and display 3-dimensional (3D) head posture in real-time. Using a mechanical head posture measuring device as a reference, 3D head positions of an artificial head and 12 human subjects were recorded with the digital head posture device. Accuracy of the digital device outputs, relationship between digital outputs and actual head rotations, and repeatability of the tests were analyzed. RESULTS: The digital head posture device showed consistent outcomes when compared to the mechanical one. The digital outputs of 3D rotations are very close to actual artificial head and human head rotations. The correlation coefficients of the linear relationship between the digital outputs and actual head movements were greater than 0.99. Repeatability tests for the artificial head and human subjects for all 3D rotations had 95% limits of agreement angles less than +/-6 degrees and +/-8 degrees, respectively. CONCLUSIONS: The digital head posture device is an acceptable device with high accuracy, repeatability, and validity in measuring head posture in 3 dimensions.