Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 44
Filter
Add more filters










Publication year range
1.
Sci Adv ; 10(16): eadj0268, 2024 Apr 19.
Article in English | MEDLINE | ID: mdl-38640247

ABSTRACT

Continuous monitoring of biomarkers at locations adjacent to targeted internal organs can provide actionable information about postoperative status beyond conventional diagnostic methods. As an example, changes in pH in the intra-abdominal space after gastric surgeries can serve as direct indicators of potentially life-threatening leakage events, in contrast to symptomatic reactions that may delay treatment. Here, we report a bioresorbable, wireless, passive sensor that addresses this clinical need, designed to locally monitor pH for early detection of gastric leakage. A pH-responsive hydrogel serves as a transducer that couples to a mechanically optimized inductor-capacitor circuit for wireless readout. This platform enables real-time monitoring of pH with fast response time (within 1 hour) over a clinically relevant period (up to 7 days) and timely detection of simulated gastric leaks in animal models. These concepts have broad potential applications for temporary sensing of relevant biomarkers during critical risk periods following diverse types of surgeries.


Subject(s)
Absorbable Implants , Transducers , Animals , Wireless Technology , Hydrogen-Ion Concentration , Biomarkers
2.
Science ; 383(6687): 1096-1103, 2024 Mar 08.
Article in English | MEDLINE | ID: mdl-38452063

ABSTRACT

Monitoring homeostasis is an essential aspect of obtaining pathophysiological insights for treating patients. Accurate, timely assessments of homeostatic dysregulation in deep tissues typically require expensive imaging techniques or invasive biopsies. We introduce a bioresorbable shape-adaptive materials structure that enables real-time monitoring of deep-tissue homeostasis using conventional ultrasound instruments. Collections of small bioresorbable metal disks distributed within thin, pH-responsive hydrogels, deployed by surgical implantation or syringe injection, allow ultrasound-based measurements of spatiotemporal changes in pH for early assessments of anastomotic leaks after gastrointestinal surgeries, and their bioresorption after a recovery period eliminates the need for surgical extraction. Demonstrations in small and large animal models illustrate capabilities in monitoring leakage from the small intestine, the stomach, and the pancreas.


Subject(s)
Absorbable Implants , Anastomotic Leak , Gastrointestinal Tract , Ultrasonics , Animals , Humans , Homeostasis , Stomach , Gastrointestinal Tract/surgery , Anastomotic Leak/diagnostic imaging , Models, Animal
3.
Science ; 383(6682): 498-503, 2024 Feb 02.
Article in English | MEDLINE | ID: mdl-38301027

ABSTRACT

The Pauson-Khand reaction has in the past 50 years become one of the most common cycloaddition reactions in chemistry. Coupling two unsaturated bonds with carbon monoxide, the transformation remains limited to CO as a C1 building block. Herein we report analogous cycloaddition reactions with nitrenes as an N1 unit. The reaction of a nonconjugated diene with a nitrene precursor produces bicyclic bioisosteres of common saturated heterocycles such as piperidine, morpholine, and piperazine. Experimental and computational mechanistic studies support relaying of the diradical nature of triplet nitrene into the π-system. We showcase the reaction's utility in late-stage functionalization of drug compounds and discovery of soluble epoxide hydrolase inhibitors.

4.
BMC Public Health ; 23(1): 2231, 2023 11 13.
Article in English | MEDLINE | ID: mdl-37957620

ABSTRACT

BACKGROUND: The increasing number of pertussis cases worldwide over the past two decades has challenged healthcare workers, and the role of environmental factors and climate change cannot be ignored. The incidence of pertussis has increased dramatically in mainland China since 2015, developing into a serious public health problem. The association of meteorological factors on pertussis has attracted attention, but few studies have examined the impact of air pollutants on this respiratory disease. METHODS: In this study, we analyzed the relationship between outdoor air pollution and the pertussis incidence. The study period was from January 2013 to December 2018, and monthly air pollutant data and the monthly incidence of patients in 31 provinces of China were collected. Distributed lag nonlinear model (DLNM) analysis was used to estimate the associations between six air pollutants and monthly pertussis incidence in China. RESULTS: We found a correlation between elevated pertussis incidence and short-term high monthly CO2 and O3 exposure, with a 10 µg/m3 increase in NO2 and O3 being significantly associated with increased pertussis incidence, with RR values of 1.78 (95% CI: 1.29-2.46) and 1.51 (95% CI: 1.16-1.97) at a lag of 0 months, respectively. Moreover, PM2.5 and SO2 also played key roles in the risk of pertussis surged. These associations remain significant after adjusting for long-term trend, seasonality and collinearity. CONCLUSIONS: Overall, these data reinforce the evidence of a link between incidence and climate identified in regional and local studies. These findings also further support the hypothesis that air pollution is responsible for the global resurgence of pertussis. Based on this we suggest that public health workers should be encouraged to consider the risks of the environment when focusing on pertussis prevention and control.


Subject(s)
Air Pollutants , Air Pollution , Whooping Cough , Humans , Incidence , Whooping Cough/epidemiology , Environmental Exposure/adverse effects , Environmental Exposure/analysis , Air Pollution/adverse effects , Air Pollution/analysis , Air Pollutants/adverse effects , Air Pollutants/analysis , China/epidemiology , Particulate Matter/adverse effects , Particulate Matter/analysis , Nitrogen Dioxide
5.
BMC Public Health ; 23(1): 2171, 2023 11 06.
Article in English | MEDLINE | ID: mdl-37932712

ABSTRACT

OBJECTIVES: To outline 44 major infectious diseases in the post-SARS (severe acute respiratory syndrome) in China and describe their long-term trends and changes by age, sex, epidemic season, and province. BACKGROUND: After the outbreak of severe acute respiratory syndrome (SARS) in 2003, with the change of infectious disease prevention and control system and the improvement of residents' quality of life, the incidence and mortality of infectious diseases have undergone major changes. METHODS: The data of 44 major infectious diseases in China from 2004 to 2018 were obtained from the monthly analysis report of the China Information System for Disease Control and Prevention (CISDCP) and the Public Health Science Data Center. Joinpoint r regression models were used to examine trends in incidence and mortality for 44 major and important infectious diseases from 2004 to 2018. RESULTS: From 2004 to 2018, 20,105, 500, 772 patients (10, 306, 546, 523 males and 9, 798, 954, 249 females) were diagnosed with 44 major infectious diseases. The overall incidence of 44 infectious diseases increased significantly from 294.6 per 100,000 people in 2004 to 479.1 per 100,000 people in 2010, with 7.9% APC (95% CI 5.2% -10.7%, P < 0.001), then slowed, and then increased to 561.2 per 100,000 people in 2018, with 1.5% APC (-0.1%-3.2%, P = 0.070). The overall mortality rose significantly, from 0.49 to 1.13 per 100,000 people between 2004 and 2011, with an APC increase of 11.6% (7.7% -15.6%, P < 0.001), and then remained stable until 2018. Among these, the prevalence of vaccine-preventable diseases and gastrointestinal & enteroviral diseases remained high and increased year by year. Patients with zoonotic diseases have the greatest risk of death, while patients with sexually transmitted and blood-borne diseases have the greatest number of deaths. Incidence rates vary considerably across geographic regions. Western China has a disproportionate burden of infectious diseases compared with eastern regions. CONCLUSIONS: After the event of SARS in 2003, infectious disease preventing and controlling model has undergone major changes in China, and certain achievements have been made in this field. Although overall morbidity and case fatality rates are still rising, they have leveled off. In reducing the disproportionate disease burden in the western region, expanding vaccination programs, preventing further increases in rates of sexually transmitted diseases, renewing efforts for emerging and persistent infectious diseases, and addressing seasonal and unpredictable outbreaks (such as the COVID-19 pandemic), there are still remain many challenges.


Subject(s)
Communicable Diseases , Pandemics , Male , Female , Humans , Quality of Life , Communicable Diseases/epidemiology , Morbidity , Incidence , China/epidemiology
6.
Environ Sci Pollut Res Int ; 30(48): 106671-106686, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37733202

ABSTRACT

Widely used agricultural greenhouses are critical in the development of facility agriculture because of not only their huge capacity in food and vegetable supplies, but also their environmental and climatic effects. Therefore, it is important to obtain the spatial distribution of agricultural greenhouses for agricultural production, policy making, and even environmental protection. Remote sensing technologies have been widely used in greenhouse extraction mainly in small or local regions, while large-scale and high-resolution (~ 1-m) greenhouse extraction is still lacking. In this study, agricultural greenhouses in an important agricultural province (Shandong, China) are extracted by the combination of high-resolution remote sensing images from Google Earth and deep learning algorithm with high accuracy (94.04% for mean intersection over union over test set). The results demonstrated that the agricultural greenhouses cover an area of 1755.3 km2, accounting for 1.11% of the total province and 2.31% of total cultivated land. The spatial density map of agricultural greenhouses also suggested that the facility agriculture in Shandong has obviously regional aggregation characteristics, which is vulnerable in both environment and economy. The results of this study are useful and meaningful for future agriculture planning and environmental management.


Subject(s)
Deep Learning , Remote Sensing Technology , Agriculture/methods , Vegetables , Conservation of Natural Resources , China
7.
Biotechnol Adv ; 69: 108261, 2023 12.
Article in English | MEDLINE | ID: mdl-37741424

ABSTRACT

Production of food-related products using microorganisms in an environmentally friendly manner is a crucial solution to global food safety and environmental pollution issues. Traditional microbial modification methods rely on artificial selection or natural mutations, which require time for repeated screening and reproduction, leading to unstable results. Therefore, it is imperative to develop rapid, efficient, and precise microbial modification technologies. This review summarizes recent advances in the construction of gene editing and metabolic regulation toolkits based on the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (CRISPR-Cas) systems and their applications in reconstructing food microorganism metabolic networks. The development and application of gene editing toolkits from single-site gene editing to multi-site and genome-scale gene editing was also introduced. Moreover, it presented a detailed introduction to CRISPR interference, CRISPR activation, and logic circuit toolkits for metabolic network regulation. Moreover, the current challenges and future prospects for developing CRISPR genetic toolkits were also discussed.


Subject(s)
CRISPR-Cas Systems , Gene Editing , CRISPR-Cas Systems/genetics , Gene Editing/methods , Food
8.
Oper Neurosurg (Hagerstown) ; 25(3): 242-250, 2023 09 01.
Article in English | MEDLINE | ID: mdl-37441801

ABSTRACT

BACKGROUND: Chronic entrapment neuropathy results in a clinical syndrome ranging from mild pain to debilitating atrophy. There remains a lack of objective metrics that quantify nerve dysfunction and guide surgical decision-making. Mechanomyography (MMG) reflects mechanical motor activity after stimulation of neuromuscular tissue and may indicate underlying nerve dysfunction. OBJECTIVE: To evaluate the role of MMG as a surgical adjunct in treating chronic entrapment neuropathies. METHODS: Patients 18 years or older with cubital tunnel syndrome (n = 8) and common peroneal neuropathy (n = 15) were enrolled. Surgical decompression of entrapped nerves was performed with intraoperative MMG of the hypothenar and tibialis anterior muscles. MMG stimulus thresholds (MMG-st) were correlated with compound muscle action potential (CMAP), motor nerve conduction velocity, baseline functional status, and clinical outcomes. RESULTS: After nerve decompression, MMG-st significantly reduced, the mean reduction of 0.5 mA (95% CI: 0.3-0.7, P < .001). On bivariate analysis, MMG-st exhibited significant negative correlation with common peroneal nerve CMAP ( P < .05), but no association with ulnar nerve CMAP and motor nerve conduction velocity. On preoperative electrodiagnosis, 60% of nerves had axonal loss and 40% had conduction block. The MMG-st was higher in the nerves with axonal loss as compared with the nerves with conduction block. MMG-st was negatively correlated with preoperative hand strength (grip/pinch) and foot-dorsiflexion/toe-extension strength ( P < .05). At the final visit, MMG-st significantly correlated with pain, PROMIS-10 physical function, and Oswestry Disability Index ( P < .05). CONCLUSION: MMG-st may serve as a surgical adjunct indicating axonal integrity in chronic entrapment neuropathies which may aid in clinical decision-making and prognostication of functional outcomes.


Subject(s)
Cubital Tunnel Syndrome , Neural Conduction , Humans , Neural Conduction/physiology , Ulnar Nerve/surgery , Cubital Tunnel Syndrome/diagnosis , Cubital Tunnel Syndrome/surgery , Muscle, Skeletal , Pain
9.
ACS Synth Biol ; 12(4): 1146-1153, 2023 04 21.
Article in English | MEDLINE | ID: mdl-37014059

ABSTRACT

The metabolic burden caused by terpenoid accumulation limits the development of highly efficient microbial cell factories, which can be circumvented using exporter-mediated product secretion. Although our previous study showed that the pleiotropic drug resistance exporter (PDR11) mediates the export of rubusoside in Saccharomyces cerevisiae, the underlying mechanism is still unclear. Herein, we used GROMACS software to simulate PDR11-mediated rubusoside recruitment and found six residues (D116, D167, Y168, P521, R663, and L1146) on PDR11 that are critical for this process. We also explored the exportation potential of PDR11 for 39 terpenoids by calculating their binding affinity using batch molecular docking. Then, we verified the accuracy of the predicted results by conducting experiments with squalene, lycopene, and ß-carotene as examples. We found that PDR11 can efficiently secrete terpenoids with binding affinities lower than -9.0 kcal/mol. Combining the computer-based prediction and experimental verification, we proved that binding affinity is a reliable parameter to screen exporter substrates and might potentially enable rapid screening of exporters for natural products in microbial cell factories.


Subject(s)
Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Terpenes/metabolism , Molecular Docking Simulation , Adenosine Triphosphate/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism
10.
Bioresour Technol ; 369: 128451, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36503088

ABSTRACT

Bioprocess control and optimization are crucial for tapping the metabolic potential of microorganisms, and which have made great progress in the past decades. Combination of the current control and optimization technologies with the latest computer-based strategies will be a worth expecting way to improve bioprocess further. Recently, artificial intelligence (AI) emerged as a data-driven technique independent of the complex interactions used in mathematical models and has been gradually applied in bioprocess. In this review, firstly, AI-guided modeling approaches of bioprocess are discussed, which are widely applied to optimize critical process parameters (CPPs). Then, AI-assisted rapid detection and monitoring technologies employed in bioprocess are summarized. Next, control strategies according to the above two technologies in bioprocess are analyzed. Lastly, current research gaps and future perspectives on AI-guided optimization and control technologies are discussed. This review provides theoretical guidance for developing AI-guided bioprocess optimization and control technologies.


Subject(s)
Artificial Intelligence , Models, Theoretical , Forecasting , Evidence Gaps
11.
J Reconstr Microsurg ; 39(3): 231-237, 2023 Mar.
Article in English | MEDLINE | ID: mdl-35952677

ABSTRACT

BACKGROUND: Commercially available near infrared spectroscopy devices for continuous free flap tissue oxygenation (StO2) monitoring can only be used on flaps with a cutaneous component. Additionally, differences in skin quality and pigmentation may alter StO2 measurements. Here, we present a novel implantable heat convection probe that measures microvascular blood flow for peripheral monitoring of free flaps, and is not subject to the same issues that limit the clinical utility of near-infrared spectroscopy. METHODS: The intratissue microvascular flow-sensing device includes a resistive heater, 4 thermistors, a small battery, and a Bluetooth chip, which allows connection to a smart device. Convection of applied heat is measured and mathematically transformed into a measurement of blood flow velocity. This was tested alongside Vioptix T.Ox in a porcine rectus abdominis myocutaneous flap model of arterial and venous occlusion. After flap elevation, the thermal device was deployed intramuscularly, and the cutaneous T.Ox device was applied. Acland clamps were alternately applied to the flap artery and veins to achieve 15 minutes periods of flap ischemia and congestion with a 15 minutes intervening recovery period. In total, five devices were tested in three flaps in three separate pigs over 16 vaso-occlusive events. RESULTS: Flow measurements were responsive to both ischemia and congestion, and returned to baseline during recovery periods. Flow measurements corresponded closely with measured StO2. Cross-correlation at zero lag showed agreement between these two sensing modalities. Two novel devices tested simultaneously on the same flap showed only minor variations in flow measurements. CONCLUSION: This novel probe is capable of detecting changes in tissue microcirculatory blood flow. This device performed well in a swine model of flap ischemia and congestion, and shows promise as a potentially useful clinical tool. Future studies will investigate performance in fasciocutaneous flaps and characterize longevity of the device over a period of several days.


Subject(s)
Free Tissue Flaps , Myocutaneous Flap , Swine , Animals , Microcirculation , Free Tissue Flaps/blood supply , Ischemia , Postoperative Complications , Arteries
12.
J Orthop Res ; 41(1): 54-62, 2023 01.
Article in English | MEDLINE | ID: mdl-35384025

ABSTRACT

Serial examination and direct measurement of intracompartmental pressure (ICP) are suboptimal strategies for the detection of acute compartment syndrome (CS) because they are operator-dependent and yield information that only indirectly reflects intracompartmental muscle perfusion. As a result, instances of unnecessary fasciotomy and unrecognized CS are relatively common. Recently, near-infrared spectroscopy (NIRS)-based systems for compartment monitoring have generated interest as an adjunct tool. Under ideal conditions, NIRS directly measures the oxygenation of intracompartmental muscle (StO2 ), thereby obviating the challenges of interpreting equivocal clinical examination or ICP data. Despite these potential advantages, existing NIRS sensors are plagued by technical difficulties that limit clinical utility. Most of these limitations relate to their transcutaneous design that makes them susceptible to both interference from intervening skin/subcutaneous tissue, underlying hematoma, and instability of the skin-sensor interface. Here, we present a flexible, wireless, Bluetooth-enabled, percutaneously introducible intramuscular NIRS device that directly and continuously measures the StO2 of intracompartmental muscle. Proof of concept for this device is demonstrated in a swine lower extremity balloon compression model of acute CS, wherein we simultaneously track muscle oxygenation, ICP, and compartment perfusion pressure (PP). The observed StO2 decreased with increasing ICP and decreasing PP and then recovered following pressure reduction. The mean change in StO2 as the PP was decreased from baseline to 30 mmHg was -7.6%. The mean difference between baseline and nadir StO2 was -17.4%. Cross-correlations (absolute value) describing the correspondence between StO2 and ICP were >0.73. This novel intramuscular NIRS device identifies decreased muscle perfusion in the setting of evolving CS.


Subject(s)
Compartment Syndromes , Spectroscopy, Near-Infrared , Swine , Animals , Muscles , Compartment Syndromes/diagnosis
13.
Comput Struct Biotechnol J ; 21: 2381-2393, 2023.
Article in English | MEDLINE | ID: mdl-38213889

ABSTRACT

Optimizing the metabolic pathways of microbial cell factories is essential for establishing viable biotechnological production processes. However, due to the limited understanding of the complex setup of cellular machinery, building efficient microbial cell factories remains tedious and time-consuming. Machine learning (ML), a powerful tool capable of identifying patterns within large datasets, has been used to analyze biological datasets generated using various high-throughput technologies to build data-driven models for complex bioprocesses. In addition, ML can also be integrated with Design-Build-Test-Learn to accelerate development. This review focuses on recent ML applications in genome-scale metabolic model construction, multistep pathway optimization, rate-limiting enzyme engineering, and gene regulatory element designing. In addition, we have discussed some limitations of these methods as well as potential solutions.

14.
Nat Commun ; 13(1): 6518, 2022 10 31.
Article in English | MEDLINE | ID: mdl-36316354

ABSTRACT

Physically transient forms of electronics enable unique classes of technologies, ranging from biomedical implants that disappear through processes of bioresorption after serving a clinical need to internet-of-things devices that harmlessly dissolve into the environment following a relevant period of use. Here, we develop a sustainable manufacturing pathway, based on ultrafast pulsed laser ablation, that can support high-volume, cost-effective manipulation of a diverse collection of organic and inorganic materials, each designed to degrade by hydrolysis or enzymatic activity, into patterned, multi-layered architectures with high resolution and accurate overlay registration. The technology can operate in patterning, thinning and/or cutting modes with (ultra)thin eco/bioresorbable materials of different types of semiconductors, dielectrics, and conductors on flexible substrates. Component-level demonstrations span passive and active devices, including diodes and field-effect transistors. Patterning these devices into interconnected layouts yields functional systems, as illustrated in examples that range from wireless implants as monitors of neural and cardiac activity, to thermal probes of microvascular flow, and multi-electrode arrays for biopotential sensing. These advances create important processing options for eco/bioresorbable materials and associated electronic systems, with immediate applicability across nearly all types of bioelectronic studies.


Subject(s)
Absorbable Implants , Electronics , Semiconductors , Electrodes , Lasers
15.
Nat Commun ; 13(1): 3040, 2022 06 01.
Article in English | MEDLINE | ID: mdl-35650215

ABSTRACT

High-sugar diet causes health problems, many of which can be addressed with the use of sugar substitutes. Rubusoside and rebaudiosides are interesting molecules, considered the next generation of sugar substitutes due to their low-calorie, superior sweetness and organoleptic properties. However, their low abundance in nature makes the traditional plant extraction process neither economical nor environmental-friendly. Here we engineer baker's yeast Saccharomyces cerevisiae as a chassis for the de novo production of rubusoside and rebaudiosides. In this process, we identify multiple issues that limit the production, including rate-liming steps, product stress on cellular fitness and unbalanced metabolic networks. We carry out a systematic engineering strategy to solve these issues, which produces rubusoside and rebaudiosides at titers of 1368.6 mg/L and 132.7 mg/L, respectively. The rubusoside chassis strain here constructed paves the way towards a sustainable, large-scale fermentation-based manufacturing of diverse rebaudiosides.


Subject(s)
Diterpenes, Kaurane , Metabolic Engineering , Diterpenes, Kaurane/metabolism , Glucosides/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Sweetening Agents
16.
Science ; 377(6601): 109-115, 2022 07.
Article in English | MEDLINE | ID: mdl-35771907

ABSTRACT

Implantable devices capable of targeted and reversible blocking of peripheral nerve activity may provide alternatives to opioids for treating pain. Local cooling represents an attractive means for on-demand elimination of pain signals, but traditional technologies are limited by rigid, bulky form factors; imprecise cooling; and requirements for extraction surgeries. Here, we introduce soft, bioresorbable, microfluidic devices that enable delivery of focused, minimally invasive cooling power at arbitrary depths in living tissues with real-time temperature feedback control. Construction with water-soluble, biocompatible materials leads to dissolution and bioresorption as a mechanism to eliminate unnecessary device load and risk to the patient without additional surgeries. Multiweek in vivo trials demonstrate the ability to rapidly and precisely cool peripheral nerves to provide local, on-demand analgesia in rat models for neuropathic pain.


Subject(s)
Absorbable Implants , Nerve Block , Neuralgia , Pain Management , Peripheral Nerves , Animals , Biocompatible Materials , Nerve Block/instrumentation , Neuralgia/therapy , Pain Management/instrumentation , Peripheral Nerves/physiopathology , Rats
17.
Nat Commun ; 13(1): 3009, 2022 05 30.
Article in English | MEDLINE | ID: mdl-35637230

ABSTRACT

Continuous, real-time monitoring of perfusion after microsurgical free tissue transfer or solid organ allotransplantation procedures can facilitate early diagnosis of and intervention for anastomotic thrombosis. Current technologies including Doppler systems, cutaneous O2-sensing probes, and fluorine magnetic resonance imaging methods are limited by their intermittent measurements, requirements for skilled personnel, indirect interfaces, and/or their tethered connections. This paper reports a wireless, miniaturized, minimally invasive near-infrared spectroscopic system designed for uninterrupted monitoring of local-tissue oxygenation. A bioresorbable barbed structure anchors the probe stably at implantation sites for a time period matched to the clinical need, with the ability for facile removal afterward. The probe connects to a skin-interfaced electronic module for wireless access to essential physiological parameters, including local tissue oxygenation, pulse oxygenation, and heart rate. In vitro tests and in vivo studies in porcine flap and kidney models demonstrate the ability of the system to continuously measure oxygenation with high accuracy and sensitivity.


Subject(s)
Oxygen Saturation , Transplants , Animals , Prostheses and Implants , Skin/diagnostic imaging , Spectroscopy, Near-Infrared/methods , Swine
18.
Microb Cell Fact ; 21(1): 74, 2022 Apr 29.
Article in English | MEDLINE | ID: mdl-35488338

ABSTRACT

BACKGROUND: Streptomyces mobaraenesis transglutaminase (smTG) is widely used to generate protein crosslinking or attachment of small molecules. However, the low thermostability is a main obstacle for smTG application. In addition, it is still hard to achieve the secretory expression of active smTG in E. coli, which benefits the enzyme evolution. In this study, a combined strategy was conducted to improve the thermostability and secretory expression of active smTG in E. coli. RESULTS: First, the thermostable S. mobaraenesis transglutaminase variant S2P-S23V-Y24N-S199A-K294L (TGm1) was intracellularly expressed in pro-enzyme form in E. coli. Fusing the pro-region of Streptomyces hygroscopicus transglutaminase (proH) and TrxA achieved a 9.78 U/mL of intracellular smTG activity, 1.37-fold higher than the TGm1 fused with its native pro-region. After in vitro activation by dispase, the TGm1 with proH yielded FRAPD-TGm1, exhibiting 0.95 â„ƒ and 94.25% increases in melting temperature and half-life at 60 â„ƒ compared to FRAP-TGm1 derived from the expression using its native pro-region, respectively. Second, the TGm1 with proH was co-expressed with transglutaminase activating protease and chaperones (DnaK, DnaJ, and GrpE) in E. coli, achieving 9.51 U/mL of intracellular FRAPD-TGm1 without in vitro activation. Third, the pelB signal peptide was used to mediate the secretory expression of active TGm in E. coli, yielding 0.54 U/mL of the extracellular FRAPD-TGm1. A script was developed to shuffle the codon of pelB and calculate the corresponding mRNA folding energy. A 1.8-fold increase in the extracellular expression of FRAPD-TGm1 was achieved by the Top-9 pelB sequence derived from the coding sequences with the lowest mRNA folding energy. Last, deleting the gene of Braun's lipoprotein further increased the extracellular yield of FRAPD-TGm1 by 31.2%, reached 1.99 U/mL. CONCLUSIONS: The stabilized FRAPD-smTG here could benefit the enzyme application in food and non-food sectors, while the E. coli system that enables secretory expression of active smTG will facilitate the directed evolution for further improved catalytic properties. The combined strategy (N-terminal modification, co-expression with chaperones, mRNA folding energy optimization of signal peptide, and lipoprotein deletion) may also improve the secretory expression of other functional proteins in E. coli.


Subject(s)
Escherichia coli , Transglutaminases , Codon , Escherichia coli/metabolism , Protein Sorting Signals/genetics , Transglutaminases/chemistry , Transglutaminases/genetics , Transglutaminases/metabolism
19.
Biosens Bioelectron ; 206: 114145, 2022 Jun 15.
Article in English | MEDLINE | ID: mdl-35278852

ABSTRACT

Vascular pedicle thrombosis after free flap transfer or solid organ transplantation surgeries can lead to flap necrosis, organ loss requiring re-transplantation, or even death. Although implantable flow sensors can provide early warning of malperfusion and facilitate operative salvage, measurements performed with existing technologies often depend on extrinsic conditions such as mounting methods and environmental fluctuations. Furthermore, the mechanisms for fixing such probes to vascular or skeletal structures may disrupt the normal blood flow or cause unnecessary tissue damage. Requirements for wired connections to benchtop readout systems also increase costs, complicate clinical care and constrain movements of the patient. Here, we report a wireless, miniaturized flow sensing system that exploits sub-millimeter scale, multi-nodal thermal probes, with biodegradable barbs that secure the probes to the surrounding tissues in a manner that facilitates removal after a period of use. These smartphone-readable devices, together with experimentally validated analytical models of the thermal transport physics, enable reliable, accurate flow sensing in ways that are largely immune to variations in temperature and mechanical perturbations. In vivo demonstrations of this technology in porcine myocutaneous flap and kidney malperfusion models highlight the essential capabilities in microsurgical and transplantation-related biomedical application scenarios.


Subject(s)
Biosensing Techniques , Transplants , Animals , Humans , Microcirculation , Prostheses and Implants , Swine
20.
J Reconstr Microsurg ; 38(4): 321-327, 2022 May.
Article in English | MEDLINE | ID: mdl-34553344

ABSTRACT

BACKGROUND: Current near-infrared spectroscopy (NIRS)-based systems for continuous flap monitoring are limited to flaps which carry a cutaneous paddle. As such, this useful and reliable technology has not previously been applicable to muscle-only free flaps where other modalities with substantial limitations continue to be utilized. METHODS: We present the first NIRS probe which allows continuous monitoring of local tissue oxygen saturation (StO2) directly within the substance of muscle tissue. This probe is flexible, subcentimeter in scale, waterproof, biocompatible, and is fitted with resorbable barbs which facilitate temporary autostabilization followed by easy atraumatic removal. This novel device was compared with a ViOptix T.Ox monitor in a porcine rectus abdominus myocutaneous flap model of arterial and venous occlusions. During these experiments, the T.Ox device was affixed to the skin paddle, while the novel probe was within the muscle component of the same flap. RESULTS: The intramuscular NIRS device and skin-mounted ViOptix T.Ox devices produced very similar StO2 tracings throughout the vascular clamping events, with obvious and parallel changes occurring upon vascular clamping and release. The normalized cross-correlation at zero lag describing correspondence between the novel intramuscular NIRS and T.Ox devices was >0.99. CONCLUSION: This novel intramuscular NIRS probe offers continuous monitoring of oxygen saturation within muscle flaps. This experiment demonstrates the potential suitability of this intramuscular NIRS probe for the task of muscle-only free flap monitoring, where NIRS has not previously been applicable. Testing in the clinical environment is necessary to assess durability and reliability.


Subject(s)
Myocutaneous Flap , Plastic Surgery Procedures , Animals , Muscles , Oxygen , Reproducibility of Results , Spectroscopy, Near-Infrared/methods , Swine
SELECTION OF CITATIONS
SEARCH DETAIL
...