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Voltage Imaging with Engineered Proton-Pumping Rhodopsins: Insights from the Proton Transfer Pathway.

Voltage imaging using genetically encoded voltage indicators (GEVIs) has taken the field of neuroscience by storm in the past decade. Its ability to create subcellular and network level readouts of electrical dynamics depends critically on the kinetics of the response to voltage of the indicator used. Engineered microbial rhodopsins form a GEVI subclass known for their high voltage sensitivity and fast response kinetics. Here we review the essential aspects of microbial rhodopsin photocycles that are critical to understanding the mechanisms of voltage sensitivity in these proteins and link the

Promotion of uterine reconstruction by a tissue-engineered uterus with biomimetic structure and extracellular matrix microenvironment.

The recurrence rate for severe intrauterine adhesions is as high as 60%, and there is still lack of effective prevention and treatment. Inspired by the nature of uterus, we have developed a bilayer scaffold (ECM-SPS) with biomimetic heterogeneous features and extracellular matrix (ECM) microenvironment of the uterus. As proved by subtotal uterine reconstruction experiments, the mechanical and antiadhesion properties of the bilayer scaffold could meet the requirement for uterine repair. With the modification with tissue-specific cell-derived ECM, the ECM-SPS had the ECM microenvironment signatu

Genetically engineered nanovesicles mobilize synergistic antitumor immunity by ADAR1 silence and PDL1 blockade.

Growing evidence has proved that RNA editing enzyme ADAR1, responsible for detecting endogenous RNA species, was significantly associated with poor response or resistance to immune checkpoint blockade (ICB) therapy. Here, a genetically engineered nanovesicle (siAdar1-LNP@mPD1) was developed as an RNA interference nano-tool to overcome tumor resistance to ICB therapies. Small interfering RNA against ADAR1 (siAdar1) was packaged into a lipid nanoparticle (LNP), which was further coated with plasma membrane extracted from the genetically engineered cells overexpressing PD1. siAdar1-LNP@mPD1 could

Emotion recognition support system: Where physicians and psychiatrists meet linguists and data engineers.

An important factor in the course of daily medical diagnosis and treatment is understanding patients' emotional states by the caregiver physicians. However, patients usually avoid speaking out their emotions when expressing their somatic symptoms and complaints to their non-psychiatrist doctor. On the other hand, clinicians usually lack the required expertise (or time) and have a deficit in mining various verbal and non-verbal emotional signals of the patients. As a result, in many cases, there is an emotion recognition barrier between the clinician and the patients making all patients seem th

Chimeric antigen receptors in the brain: Can we tackle glioblastoma with engineered NK cells?

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… Adoptive immune cell transfer is a treatment option with a long history but the administration of unmodified or cytokine-stimulated immune cells demonstrated only limited activity or was associated with toxicity. 1 More recently, engineering patient-derived immune cells has become a game changer for … A comparison between different immune effector cells, possibly also taking macrophages into consideration, will be important to define the ideal immune cell subset for genetic engineering and subsequent adoptive transfer. …

Contactless mechanical stimulation of tissue engineered constructs: Development and validation of an air-pulse device.

OBJECTIVE: Tissue engineering (TE) is the study and development of biological substitutes to restore, maintain or improve tissue function. Tissue engineered constructs (TECs) still present differences in mechanical and biological properties compared to native tissue. Mechanotransduction is the process through which mechanical stimulation triggers proliferation, apoptosis, and extracellular matrix synthesis, among other cell activities. Regarding that aspect, the effect of in vitro stimulations such as compression, stretching, bending or fluid shear stress loading modalities have been extensive…

Growth-uncoupled propanediol production in a Thermoanaerobacterium thermosaccharolyticum strain engineered for high ethanol yield.

Cocultures of engineered thermophilic bacteria can ferment lignocellulose without costly pretreatment or added enzymes, an ability that can be exploited for low cost biofuel production from renewable feedstocks. The hemicellulose-fermenting species Thermoanaerobacterium thermosaccharolyticum was engineered for high ethanol yield, but we found that the strains switched from growth-coupled production of ethanol to growth uncoupled production of acetate and 1,2-propanediol upon growth cessation, producing up to 6.7 g/L 1,2-propanediol from 60 g/L cellobiose. The unique capability of this species

Papermaking wastewater treatment coupled to 2,3-butanediol production by engineered psychrotrophic Raoultella terrigena.

The simultaneous bioremediation and bioconversion of papermaking wastewater by psychrotrophic microorganisms holds great promise for developing sustainable environments and economies in cold regions. Here, the psychrotrophic bacterium Raoultella terrigena HC6 presented high endoglucanase (26.3 U/mL), xylosidase (732 U/mL), and laccase (8.07 U/mL) activities for lignocellulose deconstruction at 15 °C. mRNA monitoring and phenotypic variation analyses confirmed that cold-inducible cold shock protein A (CspA) facilitated the expression of the cel208, xynB68, and lac432 genes to increase the enzym

Engineered Geobacillus lipolytic enzymes - Attractive polyesterases that degrade polycaprolactones and simultaneously produce esters.

Plastic pollution is one of the biggest environmental problems plaguing the modern world. Polyester-based plastics contribute significantly to this ecological safety concern. In this study, lipolytic biocatalysts GD-95RM and GDEst-lip developed based on lipase/esterase produced by Geobacillus sp. 95 strain were applied for the degradation of polycaprolactone films (Mn 45.000 (PCL45000) and Mn 80.000 (PCL80000)). The degradation efficiency was significantly enhanced by the addition of short chain alcohols. Lipase GD-95RM (1 mg) can depolymerize 264.0 mg and 280.7 mg of PCL45000 and PCL80000, fi

Engineered Stone Fabrication Work Releases Volatile Organic Compounds Classified as Lung Irritants.

Engineered stones are often characterized for their crystalline silica content. Their organic composition, particularly that of the emissions generated during fabrication work using hand-held power tools, is relatively unexplored. We forensically screened the emissions from dry-cutting 12 engineered stone products in a test chamber for their organic composition by pyrolysis-gas chromatography-mass spectrometry (GC-MS) plus selected traditional capture and analysis techniques. Phthalic anhydride, which has a Respiratory Sensitization (RSEN) Notation by the American Conference of Governmental In

Unmasking the complex roles of hypocalcemia in cancer, COVID-19, and sepsis: Engineered nanodelivery and diagnosis.

Calcium (Ca2+) homeostasis is essential for maintaining physiological processes in the body. Disruptions in Ca2+ signaling can lead to various pathological conditions including inflammation, fibrosis, impaired immune function, and accelerated senescence. Hypocalcemia, a common symptom in diseases such as acute respiratory distress syndrome (ARDS), cancer, septic shock, and COVID-19, can have both potential protective and detrimental effects. This article explores the multifaceted role of Ca2+ dysregulation in inflammation, fibrosis, impaired immune function, and accelerated senescence, contrib

Electronic Packaging Enhancement Engineered by Reducing the Bonding Temperature via Modified Cure Cycles.

Semiconductor packaging continues to reduce in thickness following the overall thinning of electronic devices such as smartphones and tablets. As the package becomes thinner, the warpage of the semiconductor package becomes more important due to the reduced bending stiffness and driven by thermal residual stresses and thermal expansion mismatch during the epoxy molding compound (EMC) curing to create the package. To address this packaging reliability issue, in this study, we developed a modified cure cycle that adds a rapid cooling step to the conventional cure cycle (CCC) to enhance the relia

Engineering a new-to-nature cascade for phosphate-dependent formate to formaldehyde conversion in vitro and in vivo.

Formate can be envisioned at the core of a carbon-neutral bioeconomy, where it is produced from CO2 by (electro-)chemical means and converted into value-added products by enzymatic cascades or engineered microbes. A key step in expanding synthetic formate assimilation is its thermodynamically challenging reduction to formaldehyde. Here, we develop a two-enzyme route in which formate is activated to formyl phosphate and subsequently reduced to formaldehyde. Exploiting the promiscuity of acetate kinase and N-acetyl-γ-glutamyl phosphate reductase, we demonstrate this phosphate (Pi)-based route in

Biological Sentinel: An Efficient Engineered Bacterial System for Monitoring and Tracing Regulated Hazardous Chemicals.

Monitoring and tracing of regulated hazardous chemicals is a public security issue of global concern. However, accurately recording historical exposure remains challenging. Here, we designed a Biological Sentinel System (BOSS) for in situ and long-term monitoring of hazardous chemical exposure using a chemical-induced base-editing system that activates antibiotic resistance screening, producing an obvious colorimetric signal. Exposure events can be written into an inheritable genomic DNA sequence, which can be read using gene sequencing. As a proof of concept, we demonstrated the accurate dete

Organ-specific model of simulated ischemia/reperfusion and hyperglycemia based on engineered heart tissue.

Here we aimed to establish an in vitro engineered heart tissue (EHT) co-morbidity mimicking model of ischemia-reperfusion injury and diabetes. EHTs were generated from primary neonatal rat cardiomyocytes. Hyperglycemic conditions or hyperosmolar controls were applied for one day to model acute hyperglycemia and for seven days to model chronic hyperglycemia. 120 min' simulated ischemia (SI) was followed by 120 min' reperfusion (R) and 1-day follow-up reperfusion (FR). Normoxic controls (N) were not subjected to SI/R. Half of the EHTs was paced, the other half was left unpaced. To assess cell in

Development of an Engineered Mycobacterium tuberculosis Strain for a Safe and Effective Tuberculosis Human Challenge Model.

Human challenge experiments could greatly accelerate the development of a tuberculosis (TB) vaccine. Human challenge for tuberculosis requires a strain that can both replicate in the host and be reliably cleared. To accomplish this, we designed Mycobacterium tuberculosis (Mtb) strains featuring up to three orthogonal kill switches, tightly regulated by exogenous tetracyclines and trimethoprim. The resultant strains displayed immunogenicity and antibiotic susceptibility similar to wild-type Mtb under permissive conditions. In the absence of supplementary exogenous compounds, the strains were ra

Investigation on Ammonia-Biodiesel Fueled RCCI Combustion Engine Using a Split Injection Strategy.

Advanced combustion concepts in compression ignition are emerging as one of the most promising solutions to reduce nitrogen oxides (NOx) and particle emissions without sacrificing fuel efficiency. Among many advanced combustion concepts, reactive controlled compression ignition (RCCI) can achieve a wider working range. In this study, to implement RCCI operation, ammonia gas is introduced through the manifold as a low-reactive fuel, and biodiesel is injected directly as a high-reactivity fuel with a 40:60 energy ratio. The effect of biodiesel split ratio in a split injection strategy (pre- and

Single-fraction Radiation Treatment Dose Response in a Genetically Engineered Mouse Model of Medulloblastoma.

Medulloblastoma is the most common malignant brain tumor of children. Although standard of care radiotherapy for pediatric medulloblastoma (PM) can lead to long-term remission or cure in many patients, it can also cause life-long cognitive impairment and other adverse effects. The pathophysiological mechanisms involved in radiation-induced cerebral damage are incompletely understood, and their elucidation may lead to interventions that mitigate radiation toxicity. To explore the mechanisms of radiation-induced cerebral damage, transgenic mouse models of PM and non-tumor-bearing controls were e

Environmental proteomics as a useful methodology for early-stage detection of stress in anammox engineered systems.

Anammox bacteria are widely applied worldwide for denitrification of urban wastewater. Differently, their application in the case of industrial effluents has been more limited. Those frequently present high loads of contaminants, demanding an individual evaluation of their treatability by anammox technologies. Bioreactors setting up and recovery after contaminants-derived perturbations are slow. Also, toxicity is frequently not acute but cumulative, which causes negative macroscopic effects to appear only after medium or long-term operations. All these particularities lead to relevant economic

Preclinical and clinical orthotopic transplantation of decellularized/engineered tracheal scaffolds: A systematic literature review.

Severe tracheal injuries that cannot be managed by mobilization and end-to-end anastomosis represent an unmet clinical need and an urgent challenge to face in surgical practice; within this scenario, decellularized scaffolds (eventually bioengineered) are currently a tempting option among tissue engineered substitutes. The success of a decellularized trachea is expression of a balanced approach in cells removal while preserving the extracellular matrix (ECM) architecture/mechanical properties. Revising the literature, many Authors report about different methods for acellular tracheal ECMs deve