Multistep Procedure Research Articles

Nanobodies Outperform Antibodies - Rapid Functionalization with Equal In Vivo Targeting Properties.

Highly specific targeting of dendritic cells in vivo is crucial for the development of effective tumor nanovaccines. This group recently presented an antibody-functionalized nanocarrier system able to maintain its targeting properties when transferred from in vitro to in vivo studies. However, producing this system requires long synthesis times and involves high expenses due to the involved site-specific enzymatic multi-step modification procedure of the antibody. Consequently, improving the previously proposed system is necessary in order to accelerate the development. Here, a novel system utilizing nanobodies for the targeting of dendritic cells is presented. A C-terminal cysteine tag facilitates an easy attachment of the nanobody to the nanocarrier via a thiol-maleimide conjugation technique. This reduces the functionalization time from several days to mere hours. Using in vitro and in vivo assays, it is shown that the optimized system possesses equal targeting properties as the antibody-based system. As a result, nanobodies and the coupling chemistry are found to be a superior strategy for the in vivo targeting of dendritic cells when compared to antibodies, due to their rapid attachment to nanocarriers and equal targeting specificity. This would replace antibodies as the current "gold standard" of targeting moieties.

Development of the scoliosis research society spinal deformity surgery safety checklist.

Spine deformity surgery is a complex multi-step procedure that has a relatively high complication rate. The use of surgical safety checklists has been shown to reduce perioperative adverse events, but existing lists are varied and non-specific for spinal deformity surgery. Thus, the purpose of this study was to develop a comprehensive surgical checklist for complex spinal corrective surgery. An electronic survey consisting of 187 surgical checklist items that had been developed and used by a group of SRS members over a 5-year period was distributed to the Scoliosis Research Society Safety and Value Committee membership. The survey sections included: (1) pre-operative area, (2) initial operating room visit, (3) before turning, (4) positioning, (5) prepare and drape, (6) pre-incision timeout, (7) intraoperative, (8) finishing implant placement and confirming imaging, (9) final rods and locking, (10) prior to closure, (11) closure, (12) turn to supine, and (13) checkout/debriefing. Respondents graded each item on a five-point Likert scale based on their perceived importance and feasibility for inclusion in the checklist. Features graded as "moderately important" or "very important" to include by at least 70% of respondents were considered to meet the cutoff for inclusion-based standard Delphi practices. Study data were collated using REDCap. A total of 25 surgeons completed the survey in its entirety. The overall checklist "package" was shortened to 9 individual checklist modules, with 2 to 16 items per checklist. In terms of individual checklist items, 40% of items (74 of 187) met the cutoff for inclusion; 17 of these items were graded as "very important," which included verifying the presence of implantable devices, reviewing the surgical plan and positioning with the surgical staff, securing the endotracheal tube, bite block confirmation, prone and lateral positioning, neuromonitoring baseline readings, double-checking that the implant screw caps were locked prior to closure, and confirming that the patient was moving bilateral lower extremities before leaving the operating room when possible. This study has led to the development of a specific spinal deformity surgical checklist of 74 (many specific to spine surgery) items that were considered important for inclusion; 17 were considered "very important".

Developing a DNA Methylation Signature to Differentiate High-Grade Serous Ovarian Carcinomas from Benign Ovarian Tumors.

Epithelial ovarian cancer (EOC) represents a significant health challenge, with high-grade serous ovarian cancer (HGSOC) being the most common subtype. Early detection is hindered by nonspecific symptoms, leading to late-stage diagnoses and poor survival rates. Biomarkers are crucial for early diagnosis and personalized treatment OBJECTIVE: Our goal was to develop a robust statistical procedure to identify a set of differentially methylated probes (DMPs) that would allow differentiation between HGSOC and benign ovarian tumors. Using the Infinium EPIC Methylation array, we analyzed the methylation profiles of 48 ovarian samples diagnosed with HGSOC, borderline ovarian tumors, or benign ovarian disease. Through a multi-step statistical procedure combining univariate and multivariate logistic regression models, we aimed to identify CpG sites of interest. We discovered 21 DMPs and developed a predictive model validated in two independent cohorts. Our model, using a distance-to-centroid approach, accurately distinguished between benign and malignant disease. This model can potentially be used in other types of sample material. Moreover, the strategy of the model development and validation can also be used in other disease contexts for diagnostic purposes.

In situ anchoring of Cu single-atom sites into porous organic polymers for efficient overall CO2 photoreduction

Single-atom site (SAS) catalysts have been extensively investigated for their remarkable catalytic properties. However, the multistep preparation procedures are complex and difficult to control. Here, we developed a one-step in situ synthesis of Cu single atom sites anchored into two types of alkynyl-linked covalent phenanthroline frameworks (CPFs) using a facile and controllable strategy during the synthesis process, that is Cu-SASs/CPF-DBPhen and Cu-SASs/CPF-TPP. Notably, the phenanthroline ligands in Cu-SASs/CPF-DBPhen have higher opening degree than the porphyrin ligands in Cu-SASs/CPF-TPP. This allows Cu-SASs/CPF-DBPhen to achieve higher metal loading and faster charge transfer efficiency than Cu-SASs/CPF-TPP, suggesting that Cu-SASs/CPF-DBPhen may have better photocatalytic performance. As expected, the CO yield of Cu-SASs/CPF-DBPhen (∼21.1 μmol·g−1·h−1) is about 11.7 times higher than that of CPF-DBPhen (∼1.8 μmol·g−1·h−1) and also exceeds that of Cu-SASs/CPF-TPP (∼8.3 μmol·g−1·h−1). Likewise, similar results were observed during the oxidation process. This work proposes a versatile one-step, in situ growth method for synthesizing catalysts.

WITHDRAWN: Characterization of Ag2O nanorods/TiO2 nanotubes hybrid coating for potential orthopedic implant applications

In this study, the potential of nanorod/nanotube hybrid arrays for orthopedic coatings was explored. The Ag2O nanorods/TiO2 nanotubes (ANRs/TNTs) hybrid coating was synthesized on Ti6Al7Nb using a multi-step procedure involving primary anodization, annealing, and secondary anodization processes. The process began with primary anodization, fostering the growth of TNTs, succeeded by thermal treatment for crystallization, and deposition of an Ag layer using physical vapor deposition (PVD) magnetron sputtering. Subsequently, secondary anodization was conducted on Ti6Al7Nb, which was initially coated with Ag-deposited TNTs, culminating in the formation of a hybrid coating. In the hybrid coating, the mean inner diameter of the TNTs was measured at 32.2 ± 8.0 nm, while the average inner diameter of the ANRs stood at 59.0 ± 30.6 nm. The hybrid coating exhibited notably higher hardness values (75.8 ± 3.2 HV) compared to both anodic TNTs (HV 45.5 ± 1.0) and crystallized TNTs (HV 54.0 ± 1.6), while its adhesion strength reached 871.5 ± 3.86 MPa, highlighting robust bonding with Ti6Al7Nb. In vitro bioactivity results confirmed bone-like apatite layer formation on ANRs/TNTs hybrid coating after 14 days in simulated body fluid (SBF), indicating enhanced bioactivity. These findings open promising avenues for advanced orthopedic coatings, with potential applications in facilitating osseointegration, drug delivery, and localized treatments in future biomedical applications.

Autologous Cryopreserved Adipose Tissue Using an Innovative Technique: An In Vitro Biological Characterization

Abstract Background Utilization of autologous adipose tissue transplantation in plastic and orthopedic surgery such as breast reconstruction and intra-articular injection has become an attractive surgical treatment with satisfactory clinical outcomes. Nevertheless, repeated liposuctions necessary to harvest fatty tissue, normally performed with sedation or general anesthesia, may represent a noteworthy concern. Objectives The aim of this study was to demonstrate through an in vitro characterization the validity of the surgical option of cryopreserved autologous adipose tissue harvested in a single shot for repeated graft transfer in breast reconstruction without impairment of cell viability and sterility. Methods Adipose tissue was collected by standard liposuction from patients who needed numerous fat grafting procedures for breast reconstruction. According to an innovative and patented cryopreservation method, autologous adipose tissue was subsequently fractioned in a sterile bag system and frozen at the RER Tissue Bank of the Emilia Romagna Region. Each graft was evaluated for sterility and cell viability immediately after harvesting, and 1, 3, 6, 12, and preliminarily 18 months after cryopreservation and thawing. Results In vitro results showed that after processing, middle-term and long-term cryopreservation, and subsequent thawing, autologous cryopreserved adipose tissue retained absence of bacterial contamination, high cellular viability, and unmodified histomorphological properties, thereby ensuring maintenance of the stromal vascular niche and the filling properties in different multistep surgical procedures. Conclusions In vitro study and sterility assessment showed that autologous cryopreserved adipose tissue grafting is a safe procedure, making it possible to avoid multiple liposuction surgery. No impairment of sterility, cell viability, or morphology was observed over time.

A one-pot, one-step CRISPR platform (CRISPR-one) for nucleic acid detection: Application for the detection of Mycoplasma pneumoniae

The conventional CRISPR-mediated detection technologies typically involve two separate steps: target pre-amplification and CRISPR-mediated signal detection. This multi-step procedure is not only cumbersome but also prone to cross-contamination. Here, we propose a new CRISPR-mediated nucleic acid detection methodology, termed CRISPR-one (CRISPR-mediated testing in one-tube, one-pot). At a constant temperature, CRISPR-one carries out both multiple cross displacement amplification (MCDA) for target pre-amplification and CRISPR/AapCas12b-based cleavage for signal detection in a single reaction pot. The novel CRISPR-one technique is employed for detecting Mycoplasma pneumoniae (M. pneumoniae), a causative agent for community-acquired pneumonia in children. M. pneumoniae CRISPR-one targets the CARDS (community-acquired respiratory distress syndrome toxin) gene and performs at 60 ºC for 50 min with only basic instrumentation required. The results can be easily visualized through a real-time fluorescence instrument, with the entire process taking less than an hour. The lowest detection limit of M. pneumoniae CRISPR-one was 50 fg (∼55 copies) of genomic DNA per reaction, and no cross-reactivity with non-M. pneumoniae templates was observed. In clinical applications, the CRISPR-one assay demonstrated a high level of agreement with both the microfluidic chip method (Kappa value: 0.95) and real-time PCR (Kappa value: 0.93) when analyzing 135 oropharynx swab samples. There is no statistical difference in detection efficiency between our assay and other methods. Together, the CRISPR-one developed here offers a rapid, convenient, highly sensitive, and specific approach for pathogen detection (e.g., M. pneumoniae), serving as an invaluable tool for clinical diagnosis and epidemiologic surveillance, particularly in resource-limited laboratories.

Advanced regression model fitting to experimental results in case of the effect of grinding conditions on a cutting force with the planer technical knives

The use of well-fit models enables users to make better decisions and draw accurate insights, so in this article a multi-step procedure for establishing regression functions for experimental data is presented. This was inspired by the need to determine well-fitted models for the results of the study of the relationship between the input factors of the grinding process of planer knives and the measured grinding power, as well as in relation to the knife cutting force. The sharpening process was carried out under various conditions, making it possible to assess their influence on the results and select the most favorable technological parameters to prepare the knives for operation. The equations adjusted by Excel or other basic methods require additional verification if the mathematical formula and values of the model coefficients correspond to the data and the physical meaning. For this purpose, one-, two- and three-factor analyses were performed to eliminate redundant factors and introduce significant interactions between input parameters. Additional analyses of variance and Tukey, Levene and Brown-Forsythe tests were used to determine the general form of the model. Detailed form of nonlinear models was worked out by comparing the distribution of residuals, looking for outliers and evidence of the presence or absence of collinearity, as well as by determining the values of additional model quality indicators, such as Akaike Information Criterion, Cook’s distance and Variance Inflation Factor. The models presented include a categorization variable (grinding type) and are presented graphically in the form of surface plots. With reference to the technological processes studied, a discussion of the physical significance of the developed models and the influence of the input factors on the model output value was conducted. The analyzes additionally include an evaluation of the model factors using prediction slice plots.

Developing a framework for identifying risk factors and estimating direct economic disease burden attributable to healthcare-associated infections: a case study of a Chinese Tuberculosis hospital

Healthcare-associated infections (HAIs) represent a major global health burden, which necessitate effective frameworks to identify potential risk factors and estimate the corresponding direct economic disease burden. In this article, we proposed a framework designed to address these needs through a case study conducted in a Tuberculosis (TB) hospital in Hubei Province, China, using data from 2018 to 2019. A comprehensive multistep procedure was developed, including ethical application, participant inclusion, risk factor identification, and direct economic disease burden estimation. In the case study, ethical approval was obtained, and patient data were anonymized to ensure privacy. All TB hospitalized patients over the study period were included and classified into groups with and without HAIs after screening the inclusion and exclusion criteria. Key risk factors, including gender, age, and invasive procedure were identified through univariate and multivariate analyses. Then, propensity score matching was employed to select the balanced groups with similar characteristics. Comparisons of medical expenditures (total medical expenditure, medicine expenditure, and antibiotics expenditure) and hospitalization days between the balanced groups were calculated as the additional direct economic disease burden measures caused by HAIs. This framework can serve as a tool for not only hospital management and policy-making, but also implementation of targeted infection prevention and control measures. Moreover, it has the potential to be applied in various healthcare settings at local, regional, national, and international levels to identify high-risk areas, optimize resource allocation, and improve hospital management and governance, as well as inter-organizational learning. Challenges to implement the framework are also raised, such as data quality, regulatory compliance, considerations on unique nature of communicable diseases and other diseases, and training need for professionals.

Multistep procedure for estimating non-linear soil response in low seismicity areas—a case study of Lucerne, Switzerland

SUMMARY The impact of non-linear soil behaviour on seismic hazard in low-to-moderate seismicity areas is often neglected; however, it may become relevant for long return periods. In this study, we used fully non-linear 1-D simulations to estimate the site-specific non-linear soil response in the low seismicity area, using the city of Lucerne in Switzerland as an example. The constitutive model considers the development of pore pressure excess and requires calibration of complex soil models, including the soil dilatancy parameters. In the absence of laboratory measurements, we mainly used the cone penetration test data to estimate the model variables and perform inversion for the dilatancy parameters. Our findings, using Swiss building code-compatible input ground motions, suggest a high probability of strong non-linear behaviour and the possibility of liquefaction at high ground motion levels in the case study area. While the non-linearity observations from strong-motion recordings are not available in Lucerne, the comparison with empirical data from other sites and other methods shows similarity with our predictions. Moreover, we show that the site response modelled is largely influenced by the strong pore pressure effects produced in thin sandy water-saturated layers. In addition, we demonstrate that the variability of the results due to the input motion and the soil parameters is significant, but within reasonable bounds.

Aminolysis-mediated single-step surface functionalization of poly (butyl cyanoacrylate) microbubbles for ultrasound molecular imaging

Molecular ultrasound imaging with actively targeted microbubbles (MB) proved promising in preclinical studies but its clinical translation is limited. To achieve this, it is essential that the actively targeted MB can be produced with high batch-to-batch reproducibility with a controllable and defined number of binding ligands on the surface. In this regard, poly (n-butyl cyanoacrylate) (PBCA)-based polymeric MB have been used for US molecular imaging, however, ligand coupling was mostly done via hydrolysis and carbodiimide chemistry, which is a multi-step procedure with poor reproducibility and low MB yield. Herein, we developed a single-step coupling procedure resulting in high MB yields with minimal batch-to-batch variation. Actively targeted PBCA-MB were generated using an aminolysis protocol, wherein amine-containing cRGD was added to the MB using lithium methoxide as a catalyst. We confirmed the successful conjugation of cRGD on the MB surface, while preserving their structure and acoustic signal. Compared to the conventional hydrolysis protocol, aminolysis resulted in higher MB yields and better reproducibility of coupling efficiency. Optical imaging revealed that under flow conditions, cRGD- and rhodamine-labelled MB, generated by aminolysis, specifically bind to tumor necrosis factor-alpha (TNF-α) activated endothelial cells in vitro. Furthermore, US molecular imaging demonstrated a markedly higher binding of the cRGD-MB than of control MB in TNF-α activated mouse aortas and 4T1 tumors in mice. Thus, using the aminolysis based conjugation approach, important refinements on the production of cRGD-MB could be achieved that will facilitate the production of clinical-scale formulations with excellent binding and ultrasound imaging performance.Graphical

Comparative evaluations of different surgical and non-surgical treatment methods for early invasive and micro invasive squamous cell carcinoma in the oral and maxillofacial regions: A systematic review

Objectives: The pathogenesis and progressive behavior of head, neck, oral and maxillofacial (HNOMF) squamous cell carcinoma (SCC) has been suggested to be a multistep and multifactorial procedure that necessitates epithelial hyperplasia, epithelial dysplasia, micro invasive squamous cell carcinoma (MISCC) and early invasive squamous cell carcinoma (EISCC); EISCC and MISCC might have a completely different behavior and development process. There are only a limited number of reported HNOMF cases of EISCC or MISCC. There are still no guidelines for the treatment of EISCC and MISCC lesions in the HNOMF regions. Material and Methods: This systematic review was conducted to gather all surgical and non-surgical treatments for EISCC and MISCC lesions in the HNOMF. The study question according to the PICO format was as followed: clinical and histopathological results (O) of all types of treatments (I) for patients with EISCC and MISCC lesions in HNOMF (P) compared to untreated lesions (C). Medline, Scopus, and Google Scholar were searched and the search was limited to English-language. Results: Eight clinical human studies were included. Photodynamic therapy (PDT) after topical application of methyl aminolevulinate (MAL-PDT) and topical Imiquimod 5 % cream both had remarkable outcomes. Conclusions: However, due to the very limited number of studies conducted on the treatment methods of MISCC and EISCC in the HNOMF regions, further studies are necessary to provide reliability for non-surgical treatment methods.

Rapid Single‐Step Hydrothermal Synthesis of Phase Pure VO2 (M) with Tailored Morphology

AbstractMonoclinic VO2 (M) is a material of great significance in the study of correlation effects in solids because of its reversible metal‐insulator phase transition between VO2 (M) and VO2 (R) just above the room temperature. Synthesizing pure monoclinic VO2 (M) can be a challenging task due to the formation of other polymorphs such as VO2 (A), VO2 (B), VO2 (D) and VO2 (P) etc. during the synthesis. Additionally, these synthesis techniques involve complex multistep procedures with protective gas environment and controlled reaction parameters clubbed with elongated reaction time ranging from 1 to 7 days which are major obstacles in the mass production of this material. Here we report a novel and rapid synthetic protocol of VO2 (M) for first time using diethylene triamine (DETA) as a reducing and morphology controlling agent in hydrogen peroxide medium. The detailed characterization of prepared materials confirms the formation of phase pure VO2 (M) crystalline powders. The present approach helps to tune these particles from one‐dimensional rods to two‐dimensional petals like structures.

Inhalable mRNA Nanoparticle with Enhanced Nebulization Stability and Pulmonary Microenvironment Infiltration.

The delivery of mRNA into the lungs is the key to solving infectious and intractable diseases that frequently occur in the lungs. Since inhalation using a nebulizer is the most promising method for mRNA delivery into the lungs, there have been many attempts toward adapting lipid nanoparticles for mRNA inhalation. However, conventional lipid nanoparticles, which have shown great effectiveness for systemic delivery of mRNA and intramuscular vaccination, are not effective for pulmonary delivery due to their structural instability during nebulization and their inability to adapt to the pulmonary microenvironment. To address these issues, we developed an ionizable liposome-mRNA lipocomplex (iLPX). iLPX has a highly ordered lipid bilayer structure, which increases stability during nebulization, and its poly(ethylene glycol)-free composition allows it to infiltrate the low serum environment and the pulmonary surfactant layer in the lungs. We selected an inhalation-optimized iLPX (IH-iLPX) using a multistep screening procedure that mimics the pulmonary delivery process of inhaled nanoparticles. The IH-iLPX showed a higher transfection efficiency in the lungs compared to conventional lipid nanoparticles after inhalation with no observed toxicity in vivo. Furthermore, analysis of lung distribution revealed even protein expression in the deep lungs, with effective delivery to epithelial cells. This study provides insights into the challenges and solutions related to the development of inhaled mRNA pulmonary therapeutics.

N/O co-doped poly(γ-glutamic acid)/Ni2+/melamine/chitosan nanoparticle-based hollow graphite carbon spheres for aqueous zinc-ion hybrid supercapacitors

Hollow carbon spheres (HCSs) have attracted broad attention in aqueous zinc-ion hybrid supercapacitors (ZIHSCs) owing to their distinctive properties. However, traditional methods for fabricating HCSs face limitations, including complex multistep procedures, the use of corrosive chemicals, and stringent reaction conditions. In this work, biomass-based poly(γ-glutamic acid)/Ni2+/melamine/chitosan nanoparticles were used as the precursors to fabricate N/O co-doped hollow graphite carbon spheres (HGCSs). Thanks to the appropriate hydrophilic characteristic, specific surface area, pore size distribution, and electrical conductivity, the fabricated HGCSs cathode exhibited superior electrochemical properties. The assembled HGCSs-based ZIHSCs device showed a satisfactory specific capacitance of 133.2 mAh·g−1 at a current density of 1.0 A·g−1, high energy densities of 75.2 Wh·kg−1 at 10,000 W·kg−1 and 107.9 Wh·kg−1 at 1000 W·kg−1, respectively. Additionally, the assembled HGCSs-based ZIHSCs device displayed an exceptional cycling stability, enduring up to 10,000 cycles at 0.5 A·g−1 with a capacity retention rate of 98.1 %. This work provides a facile and novel strategy to prepare superior electrochemical performance biomass-based HGCSs cathode for ZIHSCs.

Dimensionality of instructional quality in physical education. Obtaining students' perceptions using bifactor exploratory structural equation modeling and multilevel confirmatory factor analysis.

In research on instructional quality, the generic model of the three basic dimensions is an established framework, which postulates that the three dimensions of classroom management, student support and cognitive activation represent quality characteristics of instruction that can be generalized across subjects. However, there are hardly any studies that examine if the three basic dimensions model could represent a suitable approach to measure instructional quality in physical education. Based on an extended model of the basic dimensions, a measurement model of instructional quality for physical education is presented, which integrates different theoretical approaches from the fields of educational and psychological research as well as different subfields of sports science in order to test the factorial structure of the corresponding measurement model. 1,047 students from 72 seventh to ninth grade classes from different German-speaking Swiss cantons participated in the study. The conceptualization of the instrument is based on a hybrid approach that integrates generic and subject-specific characteristics. The simultaneous analysis at the individual and class level using MCFA was supplemented by more complex methodological techniques within the relatively new B-ESEM framework at the individual level. The postulated five-factor structure was initially tested using ICM-CFA and showed a good model fit (e.g., χ2/df = 2.32, RMSEA = 0.03, CFI = 0.97, TLI = 0.97, SRMR = 0.04). MCFA revealed a differential factorial structure at both levels of analysis with five factors at the individual level and four factors at the class level (e.g., χ2/df = 2.23, RMSEA = 0.03, CFI = 0.96, TLI = 0.96, SRMR within = 0.04, SRMR between = 0.10). ESEM and B-ESEM outperformed the ICM-CFA and showed an excellent model fit (B-ESEM: χ2/df = 1.19, RMSEA = 0.01, CFI = 1.00, TLI = 1.00, SRMR = 0.01). Inter-factor correlations and factor loadings are largely in line with expectations, indicating arguments for construct validity. The study represents a substantial contribution in linking physical education and the generic research on instructional quality. Overall, strong arguments for the factorial structure of the measurement model were demonstrated. The study can be interpreted as a first step in a multi-step procedure in terms of further validity arguments.

Hydrothermal conditioning of oleaginous yeast cells to enable recovery of lipids as potential drop-in fuel precursors

BackgroundLipids produced using oleaginous yeast cells are an emerging feedstock to manufacture commercially valuable oleochemicals ranging from pharmaceuticals to lipid-derived biofuels. Production of biofuels using oleaginous yeast is a multistep procedure that requires yeast cultivation and harvesting, lipid recovery, and conversion of the lipids to biofuels. The quantitative recovery of the total intracellular lipid from the yeast cells is a critical step during the development of a bioprocess. Their rigid cell walls often make them resistant to lysis. The existing methods include mechanical, chemical, biological and thermochemical lysis of yeast cell walls followed by solvent extraction. In this study, an aqueous thermal pretreatment was explored as a method for lysing the cell wall of the oleaginous yeast Rhodotorula toruloides for lipid recovery.ResultsHydrothermal pretreatment for 60 min at 121 °C with a dry cell weight of 7% (w/v) in the yeast slurry led to a recovery of 84.6 ± 3.2% (w/w) of the total lipids when extracted with organic solvents. The conventional sonication and acid-assisted thermal cell lysis led to a lipid recovery yield of 99.8 ± 0.03% (w/w) and 109.5 ± 1.9% (w/w), respectively. The fatty acid profiles of the hydrothermally pretreated cells and freeze-dried control were similar, suggesting that the thermal lysis of the cells did not degrade the lipids.ConclusionThis work demonstrates that hydrothermal pretreatment of yeast cell slurry at 121 °C for 60 min is a robust and sustainable method for cell conditioning to extract intracellular microbial lipids for biofuel production and provides a baseline for further scale-up and process integration.Graphical abstract

High-Throughput Bioprinting Method for Modeling Vascular Permeability in Standard Six-well Plates with Size and Pattern Flexibility.

Vascular permeability is a key factor in developing therapies for disorders associated with compromised endothelium, such as endothelial dysfunction in coronary arteries and impaired function of the blood-brain barrier. Existing fabrication techniques do not adequately replicate the geometrical variation in vascular networks in the human body, which substantially influences disease progression; moreover, these techniques often involve multi-step fabrication procedures that hinder the high-throughput production necessary for pharmacological testing. This paper presents a bioprinting protocol for creating multiple vascular tissues with desired patterns and sizes directly on standard six-well plates, overcoming existing resolution and productivity challenges in bioprinting technology. A simplified fabrication approach was established to construct six hollow, perfusable channels within a hydrogel, which were subsequently lined with human umbilical vein endothelial cells to form a functional and mature endothelium. The computer-controlled nature of 3D bioprinting ensures high reproducibility and requires fewer manual fabrication steps than traditional methods. This highlights VOP's potential as an efficient high-throughput platform for modeling vascular permeability and advancing drug discovery.

Operando-XRD of Electrochemical Reactors for Selective Li+ Extraction from Brines and Seawater

In times of rising demand of Li for use in Li-ion batteries (LIBs), and ensuring its supply for aluminum electrolysis, ultralight alloys, optics, synthesis, nuclear research, and pharmaceuticals, it is essential to simplify the processing of Li sources in a sustainable manner. Current extraction from ores is complex and requires large energy input while generating high amounts of waste. This causes the danger of making Li a limited resource and even more expensive.[1] A new promising way is the electrochemical extraction of Li by desalination batteries (DBs) from brines or seawater[2]. Thereby, selective extraction of ions can be promoted by the chemistry of the electrode material, e.g., by using FePO4. To further enhance the selectivity, potentiostatic application proved to be useful[3]. DBs are superior to other extraction methods in terms of energy efficiency, and waste generation. Still, varying Li+ concentrations in brines[1b], competitive intercalation of other cations with similar size - especially Na+ [2] -, and degradative effects of anions[4], H2O, and O2 [5] are major challenges to overcome for DBs with FePO4.In this contribution, we focus on the selective extraction of Li+ using FePO4 electrodes by specifically-designed multi-step potentiostatic application. Results are shown for salt solutions containing a mixture of Na+/Li+, and artificial seawater. It is vital to understand the atomic-scale processes so that the electrodes, and electrochemical parameters can be optimized. Therefore, we used operando high-energy X-ray diffraction (HEXRD) microscopy to investigate the crystal structure of FePO4 during the electrochemical process. The experiments showed that the energy barrier for Na+ (de)intercalation was higher than that of Li+ during galvanostatic cycling, i.e., FePO4 is selective for Li+. Using a multi-step potentiostatic procedure, the (de)intercalation process could be enhanced, as evident from our HEXRD results and simultaneous conductivity measurements. Our tentative interpretation is that Li+ is highly selectively (de)intercalated, although electrochemical signatures hinted on a two-step process during deintercalation, which may indicate the intercalation of Na+. To selectively deintercalate the remaining Na+ over Li+, we further optimized the applied potentials. During galvanostatic cycling in artificial seawater, the (de)intercalation potentials were at more positive and more negative potentials, respectively, likely caused by the variety of cations. We expect that our results contribute to the design of high-performance DBs and inspire the exploration of further systems based on our proposed multi-step process.

Non-biopsy Strategy for the Diagnosis of Celiac Disease in Adults: A Narrative Review.

Celiac disease (CeD) diagnosis is a complicated process, requiring a multi-step procedure and a high level of clinical knowledge. Some scientific societies, mainly from Europe and North America, have proposed appropriate guidelines for the diagnosis and management of CeD. Since duodenal biopsy is particularly challenging for children, guidelines of the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition, presented in 2012 and updated in 2020, have made it possible to avoid the biopsy in symptomatic pediatric patients with high levels of IgA anti-transglutaminase. Several parallel, similar studies in adults support the non-biopsy strategy. However, several pros and cons exist in applying such a strategy. The present narrative review reports the current evidence and the implication of omitting biopsy in the diagnosis of CeD in adults.