Category: Assessment of appropriate laboratory measurements to supplement the Crohn’s disease activity index
Nanotheranostics: A powerful next-generation solution to tackle hepatocellular carcinoma
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Immune Checkpoint LAG3 and Its Ligand FGL1 in Cancer
LAG3 is the most promising immune checkpoint next to PD-1 and CTLA-4. High LAG3 and FGL1 expression boosts tumor growth by inhibiting the immune microenvironment. This review comprises four sections presenting the structure/expression, interaction, biological effects, and clinical application of LAG3/FGL1. D1 and D2 of LAG3 and FD of FGL1 are the LAG3-FGL1 interaction domains. LAG3 accumulates on the surface of lymphocytes in various tumors, but is also found in the cytoplasm in non-small cell lung cancer (NSCLC) cells. FGL1 is found in the cytoplasm in NSCLC cells and on the surface of breast cancer cells. The LAG3-FGL1 interaction mechanism remains unclear, and the intracellular signals require elucidation. LAG3/FGL1 activity is associated with immune cell infiltration, proliferation, and secretion. Cytokine production is enhanced when LAG3/FGL1 are co-expressed with PD-1. IMP321 and relatlimab are promising monoclonal antibodies targeting LAG3 in melanoma. The clinical use of anti-FGL1 antibodies has not been reported. Finally, high FGL1 and LAG3 expression induces EGFR-TKI and gefitinib resistance, and anti-PD-1 therapy resistance, respectively. We present a comprehensive overview of the role of LAG3/FGL1 in cancer, suggesting novel anti-tumor therapy strategies.Futility in Transcatheter Aortic Valve Implantation: A Search for Clarity
Although transcatheter aortic valve implantation (TAVI) has revolutionised the landscape of treatment for aortic stenosis, there exists a cohort of patients where TAVI is deemed futile. Among the pivotal high-risk trials, one-third to half of patients either died or received no symptomatic benefit from the procedure at 1 year. Futility of TAVI results in the unnecessary exposure of risk for patients and inefficient resource utilisation forhealthcare services. Several cardiac and extra-cardiac conditions and frailty increase the risk of mortality despite TAVI. read more
Advection-enhanced kinetics in microtiter plates for improved surface assay quantitation and multiplexing capabilities
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Automation for Life Science Laboratories
The automation of processes in all areas of the life sciences will continue to increase in the coming years due to an ever increasing number of samples to be processed Gentaur Labware, an increasing need to protect laboratory personnel from infectious material and increasing cost pressure. Depending on the requirements of the respective application, different concepts for automation systems are available, which have a different degree of automation with regard to data handling, transportation tasks, and the processing of the samples.- Robots form a central component of these automation concepts. Classic stationary robots from the industrial sector will increasingly be replaced by new developments in the field of light-weight robots.
- In addition, mobile robots will also be of particular importance in the automation of life science laboratories in the future, especially for transportation tasks between different manual and (partially) automated stations.
- With an increasing number of different, highly diverse processes, the need for special devices and system components will also increase.
- This applies to both, the handling of the labware and the processing of the samples. In contrast to previous automation strategies with a highly parallel approach, future developments will increasingly be characterized by individual sample handling.
Fluorescence-based Single-cell Analysis of Whole-mount-stained and Cleared Microtissues and Organoids for High Throughput Screening
Three-dimensional (3D) cell culture, especially in the form of organ-like microtissues (“organoids”), has emerged as a novel tool potentially mimicking human tissue biology more closely than standard two-dimensional culture. Typically, tissue sectioning is the standard method for immunohistochemical analysis. However, it removes cells from their native niche and can result in the loss of 3D context during analyses.- Automated workflows require parallel processing and analysis of hundreds to thousands of samples, and sectioning is mechanically complex, time-intensive, and thus less suited for automated workflows.
- Here, we present a simple protocol for combined whole-mount immunostaining, tissue-clearing, and optical analysis of large-scale (approx. 1 mm) 3D tissues with single-cell level resolution.
- While the protocol can be performed manually, it was specifically designed to be compatible with high-throughput applications and automated liquid handling systems.
- This approach is freely scalable and allows parallel automated processing of large sample numbers in standard labware.
- We have successfully applied the protocol to human mid- and forebrain organoids, but, in principle, the workflow is suitable for a variety of 3D tissue samples to facilitate the phenotypic discovery of cellular behaviors in 3D cell culture-based high-throughput screens.
- Graphic abstract: Automatable organoid clearing and high-content analysis workflow and timeline.
Gold-Polyoxoborates Nanocomposite Prohibits Adsorption of Bacteriophages on Inner Surfaces of Polypropylene Labware and Protects Samples from Bacterial and Yeast Infections
Bacteriophages (phages) are a specific type of viruses that infect bacteria. Because of growing antibiotic resistance among bacterial strains, phage-based therapies are becoming more and more attractive. The critical problem is the storage of bacteriophages. Recently, it was found that bacteriophages might adsorb on the surfaces of plastic containers, effectively decreasing the titer of phage suspensions. Here, we showed that a BOA nanocomposite (gold nanoparticles embedded in polyoxoborate matrix) deposited onto the inner walls of the containers stabilizes phage suspensions against uncontrolled adsorption and titer decrease. Additionally, BOA provides antibacterial and antifungal protection. The application of BOA assures safe and sterile means for the storage of bacteriophages.Adsorption of bacteriophages on polypropylene labware affects the reproducibility of phage research
Hydrophobicity is one of the most critical factors governing the adsorption of molecules and objects, such as virions, on surfaces. Even moderate change of wetting angle of plastic surfaces causes a drastic decrease ranging from 2 to 5 logs of the viruses (e.g., T4 phage) in the suspension due to adsorption on polymer vials’ walls.- The effect varies immensely in seemingly identical containers but purchased from different vendors. Comparison of glass, polyethylene, polypropylene, and polystyrene containers revealed a threshold in the wetting angle of around 95°: virions adsorb on the surface of more hydrophobic containers, while in more hydrophilic vials, phage suspensions are stable.
- The polypropylene surface of the Eppendorf-type and Falcon-type can accommodate from around 108 PFU/ml to around 1010 PFU/ml from the suspension.
- The adsorption onto the container’s wall might result in complete scavenging of virions from the bulk. We developed two methods to overcome this issue.
- The addition of surfactant Tween20 and/or plasma treatment provides a remedy by modulating surface wettability and inhibiting virions’ adsorption.
- Plastic containers are essential consumables in the daily use of many bio-laboratories.
- Thus, this is important not only for phage-related research (e.g., the use of phage therapies as an alternative for antibiotics) but also for data comparison and reproducibility in the field of biochemistry and virology.
Resonant acoustic rheometry for non-contact characterization of viscoelastic biomaterials read more
Duohua huangjing (Polygonatum cyrtonema Hua) seedling basal rot caused by Fusarium redolens in China
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Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. Taylor, A., et al. 2016. Mol. Plant Pathol. 17:1032. https://doi.org/10.1111/mpp.12346 Fan, S. H., et al. 2021. Plant Dis. https://doi.org/10.1094/PDIS-11-19-2519-PDN Rafique, K., et al. 2020 read more
Use of equine sperm cryopreservation techniques as a conservation method of donkey germplasm
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Nitrogen resource recovery from mature leachate via heat extraction technology: An engineering project application
A large pool of ammonia in mature leachate is challenging to treat with a membrane bioreactor system to meet the discharge Standard for Pollution Control on the Landfill Site of Municipal Solid Waste in China (GB 16889-2008) without external carbon source addition. In this study, an engineering leachate treatment project with a scale of 2,000 m3/d was operated to evaluate the ammonia heat extraction system (AHES), which contains preheat, decomposition, steam-stripping, ammonia recovery, and centrifuge dewatering. The operation results showed that NH3-N concentrations of raw leachate and treated effluent from an ammonia heat extraction system (AHES) were 1,305-2,485 mg/L and 207-541 mg/L, respectively. The ratio of COD/NH3-N increased from 1.40-1.84 to 7.69-28.00. Nitrogen was recovered in the form of NH4HCO3 by the ammonia recovery tower with the introduction of CO2, wherein the mature leachate can offer 37% CO2 consumption. The unit consumptions of steam and power were 8.0% and 2.66 kWh/m3 respectively, and the total operation cost of AHES was 2.06 USD per cubic metre of leachate. These results confirm that heat extraction is an efficient and cost-effective technology for the recovery of nitrogen resource from mature leachate.Field Determination of Phosphate in Environmental Water by Using a Hand-Powered Paper Centrifuge for Preconcentration and Digital Image Colorimetric Sensing
Phosphate concentration in natural water has been used as a water quality indicator, as it is one of the major nutrients for aquatic plants. However, the traditional phosphomolybdenum blue (PMB) method has limited sensitivity for visual or camera-based detection, leading to underestimation of the phosphate concentration. We present an ultralow-cost, rapid field preconcentration and digital image colorimetric sensing of low-concentration phosphate method for water analysis. A novel hand-powered paper centrifuge (paperfuge) is used for sample preparation and preconcentration. This paperfuge is made of two circular paper discs and a string. Six centrifuge tubes (CTs) originally used as glue dispensing tips with a sample capacity of ∼230 μL, are loaded on the paperfuge. After sampling, phosphate in the water sample is reacted to form PMB. Then, the reacted sample is drawn into a CT using an autopipette before the CT bottom is sealed by glue. After Oasis HLB sorbents are added through the top of the CT, the CT top is also sealed with glue. The HLB sorbents adsorb PMB and are accumulated in the CT tip through centrifugation. The CT tips are cut and analyzed with the ImageJ software. It was found that the blue color intensity of sorbents is in a linear relationship to the phosphate concentration, with a linear range of 0-5 μM (r 2 = 0.9921) and limit of detection of 0.19 μM. In addition, this method has been applied to in-field water analysis. The results are in agreement with the standard PMB method.Pyridine-functional diblock copolymer nanoparticles synthesized via RAFT-mediated polymerization-induced self-assembly: effect of solution pH
Polymerization-induced self-assembly (PISA) via reversible addition-fragmentation chain transfer (RAFT) polymerization has become widely recognized as a versatile and efficient strategy to prepare complex block copolymer nanoparticles with controlled morphology, size, and surface functionality. In this article, we report the preparation of cationic sterically-stabilized poly(2-vinylpyridine)-poly(benzyl methacrylate) (P2VP-PBzMA) diblock copolymer nanoparticles via RAFT-mediated PISA under aqueous emulsion polymerization conditions. It is demonstrated that the solution pH during PISA has a dramatic effect on the resulting P2VP-PBzMA nanoparticles, as judged by dynamic light scattering (DLS), disc centrifuge photosedimentometry (DCP) and transmission electron microscopy (TEM). Varying the solution pH results in the P2VP stabilizer having different solubilities due to protonation/deprotonation of the pyridine groups. This allows P2VP-PBzMA nanoparticles with tunable diameters to be prepared by altering the DP of the stabilizer (P2VP) and/or core-forming block (PBzMA), or simply by changing the solution pH for a fixed copolymer composition. For example, P2VP-PBzMA nanoparticles with larger diameters can be obtained at higher solution pH as the protonation degree of the P2VP stabilizer has a large effect on both the aggregation of polymer chains during the PISA process, and the resulting behavior of the diblock copolymer nanoparticles. Changing the dispersion pH post-polymerization has a relatively limited effect on particle diameter. Furthermore, aqueous electrophoresis studies indicate that these P2VP-PBzMA nanoparticles had good colloidal stability and high cationic charge (>30 mV) below pH 5 and can be dispersed readily over a wide pH range.Simultaneous determination of 36 hypotensive drugs in fingerprints by ultra performance liquid chromatography-triple quadrupole composite linear ion trap mass spectrometry read more
Malva parviflora Leaves Mucilage: An Eco-Friendly and Sustainable Biopolymer with Antioxidant Properties
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Financial development during COVID-19 pandemic: the role of coronavirus testing and functional labs
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Recovering metal(loids) and rare earth elements from closed landfill sites without excavation: Leachate recirculation opportunities and challenges
Metal (loids) and Rare Earth Elements (REE) (‘metals’) are used in a wide range of products, and therefore, the improvement of expectations for everyday comforts with demand continues to grow. Metal-bearing wastes are a secondary source of raw material that can meet this demand by providing a previously unconsidered low impact supply source. Total annual leachate production is 1,056,716 m3. Therefore, landfill leachate emerges as a significant potential resource as it contains high concentrations of metals. However, realising a profitable return on investment for leachate processing is a challenge due to relatively low recovery rates of approximately 0.02% of total heavy metals in a landfill being leached out in 30 years. Variation within the multi-element value and the effect of other chemicals in these complex mixtures. There is a need to better understand the mechanisms and potential applicability of extraction methods for optimising metals recovery from leachate. This paper addresses this need by providing a systematic review of the critical factors and environmental conditions that influence the behaviour of metals within the landfilled waste. The paper provides a synthesis of how the factors and conditions may affect leachate recirculation efficiency for recovery in the context of a range of opportunities and challenges facing circular economy practitioners. To approach feasibility metal recovery economically from landfill leachate without energy-intensive and environmentally destructive, future research actions need to be initiated in lab-based and later on semi-pilot to pilot studies, which the review can help achieve the challenges.Amino acid pattern of rumen microorganisms in cattle fed mixed diets-An update
Rumen microorganisms turn small N-containing compounds into amino acids (AA) and contribute considerably to the supply of AA absorbed from the small intestine. Previous studies summarized the literature on microbial AA patterns, most recently in 2017 (Sok et al. Journal of Dairy Science, 100, 5241-5249). The present study intended to identify the microbial AA pattern typical when feeding Central European diets and a maximum proportion of concentrate (PCO; dry matter (DM) basis) of 0.60. Data sets were created from the literature for liquid (LAB)- and particle (PAB)-associated bacteria, total bacteria and protozoa, including 16, 9, 27 and 8 studies and 36, 21, 60 and 18 diets respectively. Because the only differences detected between LAB and PAB were slightly higher Phe and lower Thr percentages in PAB (p < 0.05), results for bacteria were pooled. A further data set evaluated AA-N (AAN) as a proportion of total N in microbial fractions and a final data set estimated protozoal contributions to total microbial N (TMN) flow to the duodenum, which were used to calculate weighted TMN AA patterns. Protozoa showed higher Lys, Asp, Glu, Ile and Phe and lower Ala, Arg, Gly, Met, Ser, Thr and Val proportions than bacteria (p < 0.05). The AAN percentage of total N in bacteria and protozoa showed large, unexplained variations, averaging 79.0% and 70.6% (p > 0.05) respectively. Estimation of protozoal contribution to TMN resulted in a cattle-specific mixed model including PCO and DM intake (DMI) per unit of metabolic body size (kg0.75 ) as fixed effects (RMSE = 3.77). With moderate PCO and DMI between 80 and 180 g/kg0.75 , which corresponds to a DMI of approximately 10 to 25 kg in a cow with 650 kg body weight, protozoal contribution ranged between 9% and 26% of TMN. Within this range, the estimated protozoal contribution to TMN resulted in minor effects on the total microbial AA pattern.Sustainable phosphorus management in soil using bone apatite
Soil fertility and phosphorus management by bone apatite amendment are receiving increasing attention, yet further research is needed to integrate the physicochemical and mineralogical transformation of bone apatite and their impact on the supply and storage of phosphorus in soil. This study has examined bone transformation in the field over a span of 10-years using a set of synchrotron-based microscopic and spectroscopic techniques. Transmission X-ray microscopy (TXM) observations reveal the in-situ deterioration of bone osteocyte-canaliculi system and sub-micron microbial tunneling within a year. Extensive organic decomposition, secondary mineral formation and re-mineralization of apatite are evident from the 3rd year. The relative ratio of (v1 + v3) PO43- to v3 CO32- and to amide I increase, and the v3c PO43- peak exhibits a blue-shift in less than 3 years. The carbonate substitution of bone hydroxyapatite (HAp) to AB-type CHAp, and phosphate crystallographic rearrangement become apparent after 10 years’ aging. The overall CO32- peak absorbance increases over time, contributing to a higher acid susceptibility in the aged bone. The X-ray Photoelectron Spectroscopy (XPS) binding energies for Ca (2p), P (2p) and O (1s) exhibit a red-shift after 1 year because of organo-mineral interplay and a blue-shift starting from the 3rd year as a result of the de-coupling of mineral and organic components. Nutrient supply to soil occurs within months via organo-mineral decoupling and demineralization. More phosphorus has been released from the bones and enriched in the associated and adjacent soils over time. Lab incubation studies reveal prominent secondary mineral formation via re-precipitation at a pH similar to that in soil, which are highly amorphous and carbonate substituted and prone to further dissolution in an acidic environment. Our high-resolution observations reveal a stage-dependent microbial decomposition, phosphorus dissolution and immobilization via secondary mineral formation over time. The active cycling of phosphorus within the bone and its interplay with adjacent soil account for a sustainable supply and storage of phosphorus nutrients.Chemical characterization of dissolved organic matter as disinfection byproduct precursors by UV/fluorescence and ESI FT-ICR MS after smoldering combustion of leaf needles and woody trunks of pine (Pinus jeffreyi) read more
Potential of cell tracking velocimetry as an economical and portable hematology analyzer
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Preparation of transparent photoluminescence smart window by integration of rare-earth aluminate nanoparticles into recycled polyethylene waste
Novel photoluminescent nanocomposite sheets were prepared for simple commercial manufacturing of transparent and luminous photochromic smart windows. Simple physical integration of lanthanide-doped strontium aluminium oxide (LdSAO) nanoparticles into recycled polyethylene (PE) waste introduced smart nanocomposite with persistent phosphorescence and photochromic properties. Because of the nanoparticle form of LdSAO is significant to develop transparent materials; LdSAO nanoparticles were well-dispersed in the polyethylene matrix. Both morphologies and chemical compositions of LdSAO nanoparticles and LdSAO-containing luminescent polyethylene sheets were investigated. Both LdSAO-free and photoluminescent polyethylene sheets were colorless in regular daylight. Only LdSAO-containing polyethylene luminescent samples showed a brilliant green color under an UV supply and greenish-yellow color under darkness as verified by CIE Lab parameters. Both absorbance and emission bands were monitored at 377 and 436/517 nm, respectively. For both photoluminescence spectroscopy and mechanical properties, the LdSAO-containing polyethylene luminescent sheets were compared to the LdSAO-free sample and found to have improved scratch resistance, UV protection, and superhydrophobic activity. Based on the added amount of LdSAO, photoluminescence, decay and lifetime spectral tests showed photochromic fluorescence and long-lasting phosphorescence characteristics. PELdSAO nanocomposite sheets displayed UV protection, photostability, hydrophobicity, excellent durability as compared to the blank LdSAO-free polyethylene sheet.Behavior of nitrogen and sulfur compounds in the rice husk pellet bioscrubber and its circulation water
In this study, pellet-type biofilter media was developed with rice husk and applied in a wet scrubber system for odor removal. The lab-scale bioscrubber system was operated for 200 days to evaluate odorous gas removal (i.e., NH3, H2S, methyl mercaptan, and dimethyl sulfide), and the removal mechanism of odor gases was studied by analyzing the behavior of nitrogen and sulfur compounds in circulation water of bioscrubber system. The rice husk pellets supplied the organic carbon source and phosphoric acid necessary for microbial growth, allowing the system to continue successfully for 200 days without any maintenance technology. By analyzing the behavior of the nitrogen and sulfur compounds in the circulation water, we confirmed that the odor gas removal resulted from various mechanisms, including adsorption and biodegradation. Ammonia gas was absorbed by the rice husk pellets and accumulated in the circulation water as nitrite under conditions of sufficient alkalinity and above pH 7. Conversely, when the alkalinity and pH decreased, nitrite was rapidly converted to nitrate. However, H2S gas was oxidized to sulfate and continuously accumulated in the circulation water regardless of the pH and alkalinity. In addition, it was confirmed that the decrease in nitrate in the bioscrubber system was due to heterotrophic denitrification by the organic carbon source supply and autotrophic denitrification by sulfur gas. During the operation of the rice husk pellet bioscrubber for 8 months, under low solubility condition, more than 99% of NH3 and H2S were removed and about 85% of methyl mercaptan (MM) and dimethyl sulfide (DMS) were removed.Financial development during COVID-19 pandemic: the role of coronavirus testing and functional labs
The outbreak of the SARS-CoV-2 virus in early 2020, known as COVID-19, spread to more than 200 countries and negatively affected the global economic output. Financial activities were primarily depressed, and investors were reluctant to start new financial investments while ongoing projects further declined due to the global lockdown to curb the disease.
Evaluation of Antitermite Properties of Wood Extracts from Pongamia pinnata (L.) Pierre (Leguminosae) against Subterranean Termites
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Isolation of Halomicroarcula pellucida strain GUMF5, an archaeon from the Dead Sea-Israel possessing cellulase
A strain designated GUMF5 was isolated in Goa-India from sediments of Dead Sea-Israel and identified as haloarchaeon Halomicroarcula pellucida based on 16S rRNA gene analysis similarity value of 99.84%. Strain GUMF5 grew on mineral salts medium with 20% NaCl and 0.5% carboxymethyl cellulose-sodium (CMC-Na) as a sole source of carbon and produced haloextremozyme cellulase. The enzyme was concentrated using Sephadex G20, precipitated with ethanol, dialyzed and retentate purified using Sephadex G200, the size exclusion chromatography. A yield of 78.53% cellulase with an activity of 131.13 U/mg and 1.24-fold purity was obtained. The purified cellulase had optimum activity at 20% NaCl, at 40 ºC, 0.5% CMC-Na, pH 7 and 150 rpm. SDS-PAGE combined with zymographic analysis revealed the molecular weight of cellulase as 240 kDa, 40 kDa and 17.4 kDa. The activity of the enzyme was stimulated by metallic cations in the order of Ca+2 > Mn+2 > Mg+2 > SO4 2- > NH4 + and was inhibited by Ag+ > Fe+2 > Cu+2. Methanol and ethanol enhanced the cellulase activity by 6% and 26%, respectively. The haloextremozyme cellulase degraded Whatman No. 1 filter paper indicated in scanning electron micrographs, exposure of open pores and fibers without any intra connectivity corresponding to paperase activity and implicating the possible use of enzyme to bio-convert cellulosic waste. Conclusively, Halomicroarcula pellucida GUMF5 (Accession number: MH244431), globally, is the only Halomicroarcula pellucida isolated from the sediments of Dead Sea producing haloextremozyme cellulase, and hence is an important biotechnological resource.Functional Comparison of Bioactive Cellulose Materials Incorporating Engineered Binding Proteins
Whatman No. 1 chromatography paper is widely used as a substrate for cellulose-based immunoassays. The immobilized proteins are used to capture target biomarkers for detection. However, alternative paper substrates may facilitate mass production of immunoassays as diagnostic tests. Here, we assessed the physical characteristics and protein immobilization capabilities of different commercial papers. Some substrates fulfilled our design criteria, including adequate flow rate and sufficient protein immobilization for efficient target capture. This study demonstrates that a variety of paper substrates can be bioactivated and used to capture target biomarkers, enabling development of affordable diagnostic tests from a range of starting materials.A practical method for storage, preservation and transportation of anuran urine samples using filter paper for hormone analysis
Anurans (frogs and toads) expelled urine when handled and it could provide insights into their physiological status. However, storage, preservation and transportation are often challenging. The study aimed to standardize and validate a field method for short-term storage and preserve of anuran urine samples using Whatman filter papers. To examine the efficacy of storage conditions and type of papers, urinary based enzyme immunoassays were used to measure progesterone and testosterone hormone metabolites.- High-Performance Liquid Chromatography was performed and revealed immunoreactive progesterone and testosterone metabolites in the urine samples.
- Urinary hormone metabolites concentration stored in filter paper at room temperature and control samples stored in -20°C for the same period were similar.
- Whatman grade 50 was found to be more suitable for storage of hormones than grade 3 paper for the experiments performed.
- A cheap and simple storage method for storage of anuran urine in field conditions using filter papers.•Anuran urine could be preserved and transported under ambient conditions without significant changes and loss of hormones.•This method would facilitate the endocrine monitoring of anurans in remote areas where limited logistics are available.
Evaluating performance of multiplex real time PCR for the diagnosis of malaria at elimination targeted low transmission settings of Ethiopia read more
Delivery Strategies for mRNA Vaccines
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Understanding the role of microperimetry in glaucoma
The present narrative review attempts to provide an overview on the use of microperimetry or fundus-driven perimetry in glaucoma, considering the clinical use, the different strategies and limits compared to standard automated perimetry. An electronic database (PubMed and Medline) search was performed of articles of any type published in the English language between 1998 and 2020 with a combination of the following terms: microperimetry, glaucoma, primary open-angle chronic glaucoma, visual field, Humphrey visual field, fundus automated perimetry. All the original articles, case reports, and short series analyzed were included in the present review, offering an excursus on the strengths and limitations characterizing the use of microperimetry in glaucomatous patients. The characteristics of a recently introduced fundus-driven perimetry Compass (CMP; Centervue, Padua, Italy) were also included. Although there remain several contradictions regarding routine use of microperimetry and the restricted research on this topic limits our ability to draw firm conclusions, microperimetry may be preferable in cases of localized retinal nerve fiber layer defects in patients with primary open-angle glaucoma and normal visual field. However, standard automated perimetry remains the gold standard for monitoring glaucoma, especially in patients with diffuse retinal nerve fiber layer impairment and visual field defects. The newly introduced Compass device can potentially provide a more accurate structural-functional evaluation than standard automated perimetry and can therefore produce superior testing reliability.Serine/Threonine-Protein
Ligand-Directed GPCR Antibody Discovery
Developing affinity reagents recognizing and modulating G-protein coupled receptors (GPCR) function by traditional animal immunization or in vitro screening methods is challenging. Some anti-GPCR antibodies exist on the market, but the success rate of development is still poor compared with antibodies targeting soluble or peripherally anchored proteins.
- More importantly, most of these antibodies do not modulate GPCR function. The current pipeline for antibody development primarily screens for overall affinity rather than functional epitope recognition. We developed a new strategy utilizing natural ligand affinity to generate a library of antibody variants with an inherent bias toward the active site of the GPCR.
- Instead of using phage libraries displaying antibodies with random CDR sequences at polymorphism sites observed in natural immune repertoire sequences, we generated focused antibody libraries with a natural ligand encoded within or conjugated to one of the CDRs or the N-terminus.
- To tailor antibody binding to the active site, we limited the sequence randomization of the antibody in regions holstering the ligand while leaving the ligand-carrying part unaltered in the first round of randomization. With hits from the successful first round, the second round of randomization of the ligand-carrying part was then performed to eliminate the bias of the ligand.
- Based on our results on three different GPCR targets, the proposed pipeline will enable the rapid generation of functional antibodies (both agonists and antagonists) against high-value targets with poor function epitope exposures including GPCR, channels, transporters as well as cell surface targets whose binding site is heavily masked by glycosylation.
endohedral trihedral metallo-borospherenes with spherical aromaticity
It is well-known that transition-metal-doping induces dramatic changes in the structures and bonding of small boron clusters, as demonstrated by the newly observed perfect inverse sandwich D8h [La(η8-B8)La] and D9h [La(η9-B9)La]–. Based on extensive global minimum searches and first-principles theory calculations, we predict herein the possibility of perfect endohedral trihedral metallo-borospherene D3h La@[La5&B30] (1, 3A’1) and its monoanion C La@[La5&B30]– (2, 2A’) and dianion D3h La@[La5&B30]2- (3, 1A’1). read more
Assessment of appropriate laboratory measurements to supplement the Crohn’s disease activity index
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