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Exosomes are extracellular vesicles with a diameter ranging from 30 to 150 nm, which are an important medium for intercellular communication and are closely related to the progression of certain diseases. Therefore, exosomes are considered promising biomarkers for the diagnosis of specific diseases, and thereby, treatments based on exosomes are being widely examined. For exosome-related research, a rapid, simple, high-purity, and recovery isolation method is the primary prerequisite for exosomal large-scale application in medical practice. Although there are no standardized methods for exosome separation and analysis, various techniques have been established to explore their biochemical and physicochemical properties. In this review, we analyzed the progress in exosomal isolation strategies and proposed our views on the development prospects of various exosomal isolation techniques.

Extracellular vesicles (EVs) include a variety of nanosized vesicles released to the extracellular microenvironment by the vast majority of cells transferring bioactive lipids, proteins, mRNA, miRNA or non-coding RNA, as means of intercellular communication. Remarkably, among other fields of research, their use has become promising for immunomodulation, tissue repair and as source for novel disease-specific molecular signatures or biomarkers. However, a major challenge is to define accurate, reliable and easily implemented techniques for EV isolation due to their nanoscale size and high heterogeneity. In this context, differential ultracentrifugation (dUC) has been the most widely used laboratory methodology, but alternative procedures have emerged to allow purer EV preparations with easy implementation. Here, we present and discuss the most used of the different EV isolation methods, focusing on the increasing impact of size exclusion chromatography (SEC) on the resulting EV preparations from in vitro cultured cells-conditioned medium and biological fluids. Comparatively, low protein content and cryo-electron microscopy analysis show that SEC removes most of the overabundant soluble plasma proteins, which are not discarded using dUC or precipitating agents, while being more user friendly and less time-consuming than gradient-based EV isolation. Also, SEC highly maintains the major EVs’ characteristics, including vesicular structure and content, which guarantee forthcoming applications. In sum, together with scaling-up possibilities to increase EV recovery and manufacturing following high-quality standards, SEC could be easily adapted to most laboratories to assist EV-associated biomarker discovery and to deliver innovative cell-free immunomodulatory and pro-regenerative therapies.

The excessive use of digital platforms with rapidly increasing users in the wireless domain enforces communication systems to provide information with high data rates, high reliability and strong transmission connection quality. Wireless systems with single antenna elements are not able to accomplish the desired needs. Therefore, multiple-input multiple-output (MIMO) antennas are getting more attention in modern high-speed communication systems and play an essential part in the current generation of wireless technology. However, along with their ability to significantly increase channel capacity, it is a challenge to achieve an optimal isolation in a compact size for fifth-generation (5G) terminals. Portable devices, automobiles, handheld gadgets, smart phones, wireless sensors, radio frequency identification and other applications use MIMO antenna systems. In this review paper, the fundamentals of MIMO antennas, the performance parameters of MIMO antennas, and different design approaches and methodologies are discussed to realize the three most commonly used MIMO antennas, i.e., ultra-wideband (UWB), dual-band and circularly polarized antennas. The recent MIMO antenna design approaches with UWB, dual band and circularly polarized characteristics are compared in terms of their isolation techniques, gain, efficiency, envelope correlation coefficient (ECC) and channel capacity loss (CCL). This paper is very helpful to design suitable MIMO antennas applicable in UWB systems, satellite communication systems, GSM, Bluetooth, WiMAX, WLAN and many more. The issues with MIMO antenna systems in the indoor environment along with possible solutions to improve their performance are discussed. The paper also focuses on the applications of MIMO characteristics for future sixth-generation (6G) technology.

George Shull’s 1908 seminal article ‘The composition of a field of maize’ marked the ‘exploitation of heterosis in plant breeding, surely one of genetics’ greatest triumphs’. Hybrid corn became a ‘symbol of American agriculture’ and ‘the paradigm for all developments of \\[ F_1\\] hybrid crop varieties and more generally breeding. But there is still no consensus on the definition of heterosis while its biological basis, causal factors and genetic mechanisms remain ‘unknown’, or at best ‘poorly understood’. It is thus logical to reverse the usual approach from the exploitation of a mysterious heterosis to the triumph of hybrid corn and focus on what breeders and geneticists do rather than on the theoretical reasons for their success. This factual approach produces surprising results: (i) hybrid corn extends the isolation technique of autogamous cereals to the allogamous maize; (ii) a ‘hybrid’ is an ordinary corn plant made reproducible by the breeder and only the breeder. It is proprietary rather than ‘hybrid’; (iii) for all practical purposes, heterosis is irrelevant; (iv) Shull justified his ‘hybrid’ breeding method by the ad hoc argument of maize ‘hybrid vigour’ which in 1914, he conflated under the name of heterosis with Edward East’s concept of physiological stimulation due to heterozygosity; (v) hybrid corn can increase yield only once and by a small margin and (vi) the huge yield gains of the last 80 years came from mass selection, a process inconsistent with the theory of heterosis. In conclusion, the enduring success of ‘hybrid’ corn was achieved at the expense of farmers, common welfare and biodiversity and dovetails with the industrial agriculture requirements of crop uniformity and breeder monopoly over reproduction. This critical understanding of the paradigm of plant breeding could have important implications for breeders and geneticists.

Summary The tea plant is an economically important woody beverage crop. The unique taste of tea is evoked by certain metabolites, especially catechin esters, whereas their precise formation mechanism in different cell types remains unclear. Here, a fast protoplast isolation method was established and the transcriptional profiles of 16 977 single cells from 1st and 3rd leaves were investigated. We first identified 79 marker genes based on six isolated tissues and constructed a transcriptome atlas, mapped developmental trajectories and further delineated the distribution of different cell types during leaf differentiation and genes associated with cell fate transformation. Interestingly, eight differently expressed genes were found to co‐exist at four branch points. Genes involved in the biosynthesis of certain metabolites showed cell‐ and development‐specific characteristics. An unexpected catechin ester glycosyltransferase was characterized for the first time in plants by a gene co‐expression network in mesophyll cells. Thus, the first single‐cell transcriptional landscape in woody crop leave was reported and a novel metabolism pathway of catechin esters in plants was discovered.

Grain legumes are widely recognized as staple sources of dietary protein worldwide. Lentil seeds are an excellent source of plant-based proteins and represent a viable alternative to animal and soybean proteins for food processing formulations. Lentil proteins provide not only dietary amino acids but are also a source of bioactive peptides that provide health benefits. This review focuses on the current knowledge of seed protein, extraction and isolation methods, bioactive peptides, and food applications of lentil protein. Lentil is the most rapidly expanding crop for direct human consumption, and has potential for greater impact as a protein source for food processing applications. Improvements in lentil protein quality, amino acid composition, and processing fractions will enhance the nutritional quality of this rapidly expanding crop globally.

Exosomes have gained recognition in cancer diagnostics and therapeutics. However, most exosome isolation methods are time-consuming, costly, and require bulky equipment, rendering them unsuitable for point-of-care (POC) settings. Microfluidics can be the key to solving these challenges. Here, we present a double filtration microfluidic device that can rapidly isolate exosomes via size-exclusion principles in POC settings. The device can efficiently isolate exosomes from 50–100 µL of plasma within 50 min. The device was compared against an already established exosome isolation method, polyethylene glycol (PEG)-based precipitation. The findings showed that both methods yield comparable exosome sizes and purity; however, exosomes isolated from the device exhibited an earlier miRNA detection compared to exosomes obtained from the PEG-based isolation. A comparative analysis of exosomes collected from membrane filters with 15 nm and 30 nm pore sizes showed a similarity in exosome size and miRNA detection, with significantly increased sample purity. Finally, TEM images were taken to analyze how the developed devices and PEG-based isolation alter exosome morphology and to analyze exosome sizes. This developed microfluidic device is cost-efficient and time-efficient. Thus, it is ideal for use in low-resourced and POC settings to aid in cancer and disease diagnostics and therapeutics.

The RNA-based study provides an excellent indication of an organism’s gene expression profile. Obtaining high-yield and high-purity RNA from Gram-positive and acid-fast bacteria is difficult without high-end kits and facilities. We optimised effective and simple protocol for RNA isolation that is a combination of enzymatic, physical and chemical treatment to disrupt cells. We successfully isolated high quality intact total RNA with yields ranging from 23.13 ± 0.40 to 61.51 ± 0.27 µg and the 260/280 purity ratio of 1.95 ± 0.01 to 2.05 ± 0.01 from Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and Mycobacterium smegmatis. These results represents a significantly enhanced yield and purity compared to other combination of techniques which we performed. Compared to previous studies the yield obtained by this method is high for the studied organisms. Furthermore the yielded RNA was successfully used for downstream applications such as quantitative real time PCR. The described method can be easily optimised and used for various bacteria.

Exosomes are a subset of nano-sized extracellular vesicles originating from endosomes. Exosomes mediate cell-to-cell communication with their cargos, which includes mRNAs, miRNAs, lncRNAs, and circRNAs. Exosomal RNAs have cell specificity and reflect the conditions of their donor cells. Notably, their detection in biofluids can be used as a diagnostic marker for various diseases. Exosomal RNAs are ideal biomarkers because their surrounding membranes confer stability and they are detectable in almost all biofluids, which helps to reduce trauma and avoid invasive examinations. However, knowledge of exosomal biomarkers remains scarce. The present review summarizes the biogenesis, secretion, and uptake of exosomes, the current researches exploring exosomal mRNAs, miRNAs, lncRNAs, and circRNAs as potential biomarkers for the diagnosis of human diseases, as well as recent techniques of exosome isolation.