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The ability of the complement system to rapidly and broadly react to microbial intruders, apoptotic cells and other threats by inducing forceful elimination responses is indispensable for its role as host defense and surveillance system. However, the danger sensing versatility of complement may come at a steep price for patients suffering from various immune, inflammatory, age-related, or biomaterial-induced conditions. Misguided recognition of cell debris or transplants, excessive activation by microbial or damaged host cells, autoimmune events, and dysregulation of the complement response may all induce effector functions that damage rather than protect host tissue. Although complement has long been associated with disease, the prevalence, impact and complexity of complement’s involvement in pathological processes is only now becoming fully recognized. While complement rarely constitutes the sole driver of disease, it acts as initiator, contributor, and/or exacerbator in numerous disorders. Identifying the factors that tip complement’s balance from protective to damaging effects in a particular disease continues to prove challenging. Fortunately, however, molecular insight into complement functions, improved disease models, and growing clinical experience has led to a greatly improved understanding of complement’s pathological side. The identification of novel complement-mediated indications and the clinical availability of the first therapeutic complement inhibitors has also sparked a renewed interest in developing complement-targeted drugs, which meanwhile led to new approvals and promising candidates in late-stage evaluation. More than a century after its description, complement now has truly reached the clinic and the recent developments hold great promise for diagnosis and therapy alike.

Cells have different sets of molecules for performing an array of physiological functions. Nucleic acids have stored and carried the information throughout evolution, whereas proteins have been attributed to performing most of the cellular functions. To perform these functions, proteins need to have a unique conformation and a definite lifespan. These attributes are achieved by a highly coordinated protein quality control (PQC) system comprising chaperones to fold the proteins in a proper three-dimensional structure, ubiquitin-proteasome system for selective degradation of proteins, and autophagy for bulk clearance of cell debris. Many kinds of stresses and perturbations may lead to the weakening of these protective cellular machinery, leading to the unfolding and aggregation of cellular proteins and the occurrence of numerous pathological conditions. However, modulating the expression and functional efficiency of molecular chaperones, E3 ubiquitin ligases, and autophagic proteins may diminish cellular proteotoxic load and mitigate various pathological effects. Natural medicine and small molecule-based therapies have been well-documented for their effectiveness in modulating these pathways and reestablishing the lost proteostasis inside the cells to combat disease conditions. The present article summarizes various similar reports and highlights the importance of the molecules obtained from natural sources in disease therapeutics., Graphical abstract Extra-/intracellular stresses challenge cellular health throughout life. Small natural molecules modulate the functioning of protein quality control components to maintain proteins' native structures and preserve cells’ proteome under healthy and stressful conditions.Image 1

Objectives/purpose High-touch surfaces are a critical reservoir in the spread of nosocomial infections. Although disinfection and infection control protocols are well developed, they lack the ability to passively reduce the pathogenic load of high-touch surfaces. Copper and its alloys have been suggested as a surface that exhibit continuous biocidal effects. Antimicrobial studies on these surfaces are prevalent, while virucidal studies are not as well explored. The goal of this study was to first determine the virucidal activity of a copper–nickel–zinc alloy and to then examine the effect of soiling and virus preparation on virucidal activity. Methods A baculovirus vector was used as an easily quantifiable model of an infectious enveloped animal cell virus. Droplets containing virus were deposited on surfaces and allowed to stay wet using humidity control or were dried onto the surface. Virus was then recovered from the surface and assayed for infectivity. To examine how the composition of the droplet affected the survival of the virus, 3 different soiling conditions were tested. The first two were recommended by the United States Environmental Protection Agency and the third consisted of cell debris resulting from virus amplification. Results A copper–nickel–zinc alloy was shown to have strong virucidal effects for an enveloped virus. Copper, nickel, and zinc ions were all shown to leach from the alloy surface and are the likely cause of virucidal activity by this surface. Virucidal activity was achieved under moderate soiling but lost under high soiling generated by routine virus amplification procedures. The surface was able to repeatably inactivate dried virus droplets under moderate soiling conditions, but unable to do so for virus droplets kept wet using high humidity. Conclusion Ion leaching was associated with virucidal activity in both wet and dried virus conditions. Soiling protected the virus by quenching metal ions, and not by inhibiting leaching. The composition of the solution containing virus plays a critical role in evaluating the virucidal activity of surfaces and surface coatings.

Background Preimplantation genetic testing (PGT) is offered to a wide range of structural and numerical chromosomal imbalances, with PGT- polymerase chain reaction (PCR), as the method of choice for amplifying the small DNA content achieved from the blastomere biopsy or trophectoderm (TE) biopsy, that might have a detrimental impact on embryonic implantation potential. Since human embryos cultured until Day-5–6 were noticed to expel cell debris/ fragments within the zona pellucida, we aimed to examine whether these cell debris/ fragments might be used for PGT, as an alternative to embryo biopsy. Methods Blastocysts, which their Day-3 blastomere biopsy revealed an affected embryo with single-gene defect, and following hatching leaved cell debris/fragments within the zona pellucida were analyzed. Each blastocyst and its corresponding cell debris/fragments were separated and underwent the same molecular analysis, based on multiplex PCR programs designed for haplotyping using informative microsatellites markers. The main outcome measure was the intra-embryo congruity of Day-3 blastomere biopsy and its corresponding blastocyst and cell debris/fragments. Results Fourteen affected embryos from 9 women were included. Only 8/14 (57.2%) of embryos demonstrated congruent molecular genetic results between Day-3 embryo and its corresponding blastocyst and cell debris/fragments. In additional 6/14 (42.8%) embryos, molecular results of the Day-3 embryos and their corresponding blastocysts were congruent, while the cell debris/fragments yielded no molecular diagnoses (incomplete diagnoses). Conclusions It might be therefore concluded, that in PGT cycles, examining the cell debris/fragments on Day-4, instead of Day-3 blastomere or Day-5 TE biopsies, is feasible and might avoid embryo biopsy with its consequent detrimental effect on embryos’ implantation potential. Whenever the latter results in incomplete diagnosis, TE biopsy should be carried out on Day-5 for final genetic results. Further large well-designed studies are required to validate the aforementioned PGT platform.

BACKGROUND The present study investigated the sensory acceptance, oxidative damage and protection, and possible anatomical-structural damage of cells from the surface of shapes of minimally processed yam. The tubers were minimally processed into the peeled rondelle, dice and 'chateau cut' (chateau) shapes, the latter of which was obtained after performing the abrasion technique. Control treatment corresponded to the rondelle shape with the periderm. The pieces were kept packed at 5 ± 2 °C for 14 days. RESULTS Peeled rondelle and chateau were sensorially the most well-accepted yam shapes and achieved the highest purchase intention. The enzymes were partially modulated by the detected H2 O2 levels. Oxidative burst lasted longer in the minimally processed tissues than in the control. Polyphenol oxidase activity showed a clear difference in behavior between the minimally processed pieces and the control. Minimal processing induced transient increases in phenolic compounds, for which the expression was lowest in the abraded pieces. On the other hand, these pieces exhibited greater cell collapse on the surface of the amyliferous parenchyma. CONCLUSION Based on the results of the trained panel, the abrasion technique is an alternative to provide shapes that are better accepted and marketable, more resistant to browning, and can be stored for up to 12 days. Resistance to browning may be related to a more efficient modulation of enzymatic antioxidant systems and intense deposition of cell debris on the surface of the amyliferous parenchyma. © 2021 Society of Chemical Industry.

Background Most laboratories use specialized tubes (e.g., Streck) to recover circulating cell-free DNA (ccfDNA) for noninvasive prenatal testing (NIPT). We validated a low cost, simple procedure for collecting NIPT samples in remote laboratories that avoids highspeed centrifugation. EDTA gel blood sampling tube allows simple separation of plasma from blood cells. Decanted plasma is filtered to remove cell debris. The procedure can be performed within a few minutes after the blood centrifugation step, and ccfDNA-grade plasma can be frozen for transportation. Methods We recruited 51 pregnant women and collected blood in one EDTA-gel Greiner tube and two Streck tubes. All tubes were centrifuged at 1600 g x 10 min within 6 h of sample collection. Plasma from EDTA tubes was poured into a syringe cylinder and filtered through a 0.45 µm Millipore filter. Plasma from Streck tubes was recovered with a pipette and one was filtered as above while the second was centrifuged at 16 000 g. The ccfDNA was isolated and NGS sequencing libraries were prepared and sequenced on an Illumina system. Fetal fractions were estimated using SeqFF. This study had a power of 79% to detect a decrease of 1% in fetal fractions with the new method. Results We did not observe any significant difference between the three procedures for the fetal fraction nor for the quality or quantity of libraries produced. Conclusion EDTA-gel tubes with filtration provide high quality plasma for ccfDNA analysis and can be sent frozen to the NIPT laboratory. This is economical and it frees the laboratory of time-consuming steps.

Myocardial infarction (MI) is an irreversible damage of the heart muscle, which often leads to adverse cardiac remodeling and progressive heart failure. After MI, immune cells play a vital role in the clearance of the dying tissue and cardiac remodeling. Post-MI events include the release of danger signals by necrotic cardiomyocytes and the migration of the inflammatory cells, such as dendritic cells, neutrophils, monocytes, and macrophages, into the site of the cardiac injury to digest the cell debris and secrete a variety of inflammatory factors activating the inflammatory response. In this review, we focus on the role of immune cells in the cardiac remodeling after MI and the novel immunotherapies targeting immune cells.

There are concepts in science that need time to overcome initial disbelief before finally arriving at the moment when they are embraced by the research community. One of these concepts is the biological meaning of the small, spheroidal vesicles released from cells, which are described in the literature as microparticles, microvesicles, or exosomes. In the beginning, this research was difficult, as it was hard to distinguish these small vesicles from cell debris or apoptotic bodies. However, they may represent the first language of cell–cell communication, which existed before a more specific intercellular cross-talk between ligands and receptors emerged during evolution. In this review article, we will use the term “extracellular microvesicles” (ExMVs) to refer to these small spheroidal blebs of different sizes surrounded by a lipid layer of membrane. We have accepted an invitation from the Editor-in-Chief to write this review in observance of the 20th anniversary of the 2001 ASH Meeting when our team demonstrated that, by horizontal transfer of several bioactive molecules, including mRNA species and proteins, ExMVs harvested from embryonic stem cells could modify hematopoietic stem/progenitor cells and expand them ex vivo. Interestingly, the result that moved ExMV research forward was published first in 2005 in Leukemia, having been previously rejected by other major scientific journals out of simple disbelief. Therefore, the best judge of a new concept is the passage of time, although the speed of its adoption is aided by perseverance and confidence in one’s own data. In this perspective article, we will provide a brief update on the current status of, hopes for, and likely future of ExMV research as well as therapeutic and diagnostic applications, with a special emphasis on hematopoiesis.

Finger-stick blood sampling is convenient for point of care diagnostics, but whole blood samples are problematic for many assays because of severe matrix effects associated with blood cells and cell debris. We introduce a new digital microfluidic (DMF) diagnostic platform with integrated porous membranes for blood-plasma separation from finger-stick blood volumes, capable of performing complex, multi-step, diagnostic assays. Importantly, the samples can be directly loaded onto the device by a finger "dab" for user-friendly operation. We characterize the platform by comparison to plasma generated via the "gold standard" centrifugation technique, and demonstrate a 21-step rubella virus (RV) IgG immunoassay yielding a detection limit of 1.9 IU mL-1, below the diagnostic cut-off. We propose that this work represents a critical next step in DMF based portable diagnostic assays-allowing the analysis of whole blood samples without pre-processing.

Since extracellular vesicles (EVs) were discovered in 1983 in sheep reticulocytes samples, they have gradually attracted scientific attention and become a topic of great interest in the life sciences field. EVs are small membrane particles, released by virtually every cell that carries a variety of functional molecules. Their main function is to deliver messages to the surrounding area in both physiological and pathological conditions. Initially, they were thought to be either cell debris, signs of cell death, or unspecific structures. However, accumulating evidence support a theory that EVs are a universal mechanism of communication. Thanks to their biological characteristics and functions, EVs are likely to represent a promising strategy for obtaining pathogen information, identifying therapeutic targets and selecting specific biomarkers for a variety of diseases, such as autoimmune diseases. In this review, we provide a brief overview of recent progress in the study of the biology and functions of EVs. We also discuss their roles in diagnosis and therapy, with particular emphasis on autoimmune diseases.

A laparoscopic approach is suggested with the highest grade of recommendation for acute cholecystitis, perforated gastroduodenal ulcers, acute appendicitis, gynaecological disorders, and non-specific abdominal pain (NSAP). To date, the main qualities of laparoscopy for these acute surgical scenarios are clearly stated: quicker surgery, faster recovery and shorter hospital stay. For the remaining surgical emergencies, as well as for abdominal trauma, the role of laparoscopy is still a matter of debate. Patients might benefit from a laparoscopic approach only if performed by experienced teams and surgeons which guarantee a high standard of care. More precisely, laparoscopy can limit damage to the tissue and could be effective for the reduction of the overall amount of cell debris, which is a result of the intensity with which the immune system reacts to the injury and the following symptomatology. In fact, these fragments act as damage-associated molecular patterns (DAMPs). DAMPs, as well as pathogen associated molecular patterns (PAMPs), are recognised by both surface and intracellular receptors of the immune cells and activate the cascade which, in critically ill surgical patients, is responsible for a deranged response. This may result in the development of progressive and multiple organ dysfunctions, manifesting with acute respiratory distress syndrome (ARDS), coagulopathy, liver dysfunction and renal failure. In conclusion, none of the emergency surgical scenarios preclude laparoscopy, provided that the surgical tactic could ensure sufficient cleaning of the abdomen in addition to resolving the initial tissue damage caused by the “trauma”.

The swim bladder inflates early after fish hatching via its interconnection with the digestive tract (ductus pneumaticus). This interconnection may serve as a portal to foreign particles, including bacteria, causing deficiencies in primary swim bladder inflation. We histologically examined 134 African annual killifish (genus Nothobranchius) with secondary loss of swim bladder function ('belly sliders'). We demonstrate that these fish lost the ability of air regulation in their swim bladders likely due to Mycobacterium spp. infection at an individual-specific age. Nearly all examined belly sliders had thickened swim bladder walls, and their swim bladder was filled with material containing mycobacteria, cell debris, young monocytic cells and phagocyting macrophages. Mycobacterial infection was restricted to the swim bladder in juveniles, where mycobacteria likely enter the host through the ductus pneumaticus. Infection in adults was systemic and mycobacteria were present in all examined organs. Presence of mycobacteria in the epithelial lining and submucosal layers of the digestive tract of adults suggests that it may also serve as the entrance site of infection. We suspect 2 sources of Mycobacterium contamination: dietary (with bloodworms) and/or contaminated hatching substrate. These sources of contamination may be eliminated by use of laboratory dry feed and egg disinfection prior to hatching.

The tumor immune microenvironment plays a vital role in the metastasis of colorectal cancer. As one of the most important immune cells, macrophages act as phagocytes, patrol the surroundings of tissues, and remove invading pathogens and cell debris to maintain tissue homeostasis. Significantly, macrophages have a characteristic of high plasticity and can be classified into different subtypes according to the different functions, which can undergo reciprocal phenotypic switching induced by different types of molecules and signaling pathways. Macrophages regulate the development and metastatic potential of colorectal cancer by changing the tumor immune microenvironment. In tumor tissues, the tumor-associated macrophages usually play a tumor-promoting role in the tumor immune microenvironment, and they are also associated with poor prognosis. This paper reviews the mechanisms and stimulating factors of macrophages in the process of colorectal cancer metastasis and intends to indicate that targeting macrophages may be a promising strategy in colorectal cancer treatment.

The interior of living cells is a molecular-crowding environment, where large quantities of various molecules coexist. Investigations into the nature of this environment are essential for an understanding of both the elaborate biological reactions and the maintenance of homeostasis occurring therein. The equilibrium states of biological macromolecular systems are affected by molecular-crowding environments unmatched by in vitro diluted environments; knowledge about crowding effects is still insufficient due to lack of relevant experimental studies. Recent developments in the techniques of in-cell NMR and large-scale molecular dynamics simulation have provided new insights into the structure and dynamics of biological molecules inside the cells. This study focused on a new experimental technique to directly observe the structure of a specific protein or membrane in condensed crowder solutions using neutron scattering. Deuterated whole-cell debris was used to reproduce an environment that more closely mimics the interior of living cells than models used previously. By the reduction of the background scattering from large amounts of cell debris, we successfully extracted structure information for both small globular protein and small unilamellar vesicle (SUV) from the concentrated cell-debris solution up to a weight ratio of 1:60 for protein/crowder and 1:40 for SUV/crowder.

1,3-diaminopropane (1,3-DAP), 1,4-diaminobutane (1,4-DAB), and 1,5-diaminopentane (1,5-DAP) are important chemicals due to their wide industrial use including agrochemicals, pharmaceuticals, surfactants, and building blocks for nylon synthesis. Over the last decade, several studies on bio-based production of diamines by metabolically engineered microorganisms have been published. However, development of efficient methods for the recovery of 1,3-DAP, 1,4-DAB, and 1,5-DAP from fermentation broth has not been reported. In this study, an efficient process for the separation and purification of 1,3-DAP, 1,4-DAB, and 1,5-DAP from fermentation broth without using highly flammable or toxic solvents was developed. The optimal process for the recovery and purification of these diamines from fermentation broth comprises several unit operations including removal of cell debris, decolorization of fermentation broth, product concentration, deprotonation of diamines, product separation, and final polishing to obtain pure 1,3-DAP, 1,4-DAB, and 1,5-DAP with yields of 87 +/- 3%, 86 +/- 4%, and 81 +/- 2%, respectively. Process development for the recovery and purification of fermentatively produced diamines reported here is expected to facilitate synthesis of bio-based nylons. Furthermore, the strategy reported here could be similarly applicable in developing downstream processes to recover and purify other diamines and related chemicals from fermentation broth. (C) 2018 Elsevier Ltd. All rights reserved.

The concept of vesicles or cell debris released by cancer cells to promote metastasis is not new, but the mechanisms used to currently ascribe their impact in metastasis are of intense debate. A significant increase in reports describing the role of cancer-derived EVs in cancer metastasis has been followed by a growing amount of uncertainty behind these claims. This review will delve into the role of EVs in promoting cancer metastasis by relying on a balanced perspective that looks at challenges faced previously by extracellular vesicle biologists, current technical limitations in the field, and overlooked physiologic mechanisms that may play a confounding role. This review will also discuss how certain experimental approaches are misleading which ultimately lead to overly optimistic mechanisms that have minimally contributed to the pathophysiology of metastasis.

In order to reduce the widespread threat of bacterial pathogen diseases, mechanical bactericidal surfaces have been widely reported. However, few of these nanostructured surfaces were investigated from a sustainable perspective. In this study, we have prepared, inspired by the slippery zone of Nepenthes, a multifunctional nanostructured surface with mechanical bactericidal, self-cleaning and insect anti-adhesive characteristics. First, a nanoblade-like surface made of Zn–Al layered double hydroxides was prepared for achieving faster bactericidal rate and wider bactericidal spectrum (2.10 × 104 CFU cm−2 min−1 against Escherichia coli and 1.78 × 103 CFU cm−2 min−1 against Staphylococcus aureus). Then the self-cleaning and insect anti-adhesive properties were tested on the fluorosilane-modified nanoblades, leaving little cell debris remaining on the surface even after 4 continuous bactericidal experiments, and showing a slippery surface for ants to slide down in 3 s. This study not only discovers a new nature-inspired mechanical bactericidal nanotopography, but also provides a facile approach to incorporate multiple functions into the nanostructured surface for practical antibacterial applications.

Preparing a single cell suspension is a critical step in any solid tissue flow cytometry experiment. Tissue dissection, enzymatic digestion, and mechanical dissociation are three significant steps leading to the degradation of the extracellular matrix and the isolation of single cells, allowing the generation of high-quality flow cytometry data. Cells and the extracellular matrix contain various proteins and other structures which must be considered when designing a tissue digestion protocol to preserve the viability of cells and the presence of relevant antigens while digesting matrix components and cleaving cell-cell junctions. Evaluation of the single cell suspension is essential before proceeding with the labeling of the cells as high viability and absence of cell debris and aggregates are critical for flow cytometry. The information presented should be used as a general guide of steps to consider when preparing a single cell suspension from solid tissues for flow cytometry experiments. PURPOSE: The purpose of this paper is to define the critical steps in the segregation of solid tissues by describing the composition of tissues and the common digestive enzymes used to digest extracellular matrix and cleave cell-cell junctions to obtain a single cell suspension.

ZusammenfassungDas Spülen von Wunden mit einer Lösung zur Wundreinigung ist seit Langem ein anerkannter Eckpfeiler in der Wundbehandlung als Mittel zur Entfernung von Zelltrümmern und Oberflächenpathogenen in Wundexsudaten. In Kombination mit dem chirurgischen Débridement und einer lokalen Vakuumtherapie kann sie das Fortschreiten von der entzündlichen zur proliferativen Phase der Wundheilung erleichtern. Verfahren der topischen Vakuumtherapie mit Instillation und einer definierten Einwirk- bzw. Verweilzeit von topischen Lösungen unter zyklischer Kompression und Dekompression mit Schaumverbänden (Negative Pressure Wound Therapy with instillation and dwell time = NPWTi-d) können Rückstände entfernen, die ansonsten hemmend auf den Heilungsprozess wirken. Gleichzeitig helfen sie, die bakterielle Keimbelastung in kontaminierten oder infizierten Wunden zu verringern. Nachdem diese Technik nun kommerziell verfügbar und zunehmend verbreitet ist, wurden im Rahmen eines Konsensusmeetings durch eine interdisziplinäre Expertenkommission Empfehlungen zur Anwendung und zu den klinischen Indikationen erarbeitet. Auch wenn die Evidenzstufe von Expertenmeinungen einen geringeren Level besitzt, können allgemein gültige Richtlinien für einen sicheren und effizienten Einsatz von NPWTi-d ausgesprochen werden, die dem klinisch tätigen Arzt als Handlungsempfehlung dienen können. Die daraus abgeleiteten Konsensusempfehlungen umfassen basierend auf dem Stand der aktuellen wissenschaftlichen Datenlage die Ziele der Behandlung, die Anwendungsmodalitäten die Indikationsstellung bei verschiedenen Wunden einschließlich eventueller Kontraindikationen, Therapieeinstellungen sowie die Verwendung topischer Instillationslösungen, deren Volumen und Verweildauer (dwell time), die optimale Behandlungsdauer und zukünftige Weiterentwicklungen der NPWTi-d.

Background:. We aimed to evaluate the use of nerve allograft preserved in glycerol. We compared the efficiency of glycerol-preserved allografts with autogenous nerve grafting, cryopreserved grafts, and detergent-processed grafts in the axonal regeneration. Secondarily, we evaluated the effectiveness of each preservation method in maintaining the extracellular matrix free of cellular components. Methods:. This was a prospective experimental, longitudinal, unblinded, nonrandomized, controlled animal model study. Three different allograft preservation techniques for the repair of sciatic nerve injuries were compared, including cold preservation, glycerol preservation, and detergent preservation. Functional assessment was performed, and histomorphometric analyses were further performed, which enabled the allograft structure evaluation and an estimation of the nerve regeneration efficacy based on the myelinated axons count and on their diameters. Results:. After the 14th week, all groups were already balanced and similar (P = 0.265): all groups present near-zero SFIs, thus confirming their efficiency in promoting nerve regeneration. In the histomorphometric evaluations, all groups were equivalent, presenting a similar efficiency in nerve regeneration (P = 0.716 and P = 0.577, respectively). Similarly, histomorphometric evaluations showed a reduction in the number of axons and in their diameters, but none of them effectively eliminated all cellular debris. Comparing the groups with each other, the groups preserved in glycerol and detergent solution were similar, both presenting better results than the cooled group. Conclusion:. By evaluating the presence of cell debris after the treatment using glycerol, it was found to be similar to the treatment using detergent and significantly better than the cold-preservation treatment.

The coronavirus disease 2019 (COVID-19) pandemic has highlighted bottlenecks in large-scale, frequent testing of populations for infections. Polymerase chain reaction (PCR)-based diagnostic tests are expensive, reliant on centralized labs, can take days to deliver results, and are prone to backlogs and supply shortages. Antigen tests that bind and detect the surface proteins of a virus are rapid and scalable but suffer from high false negative rates. To address this problem, an inexpensive, simple, and robust 60-minute do-it-yourself (DIY) workflow to detect viral RNA from nasal swabs or saliva with high sensitivity (0.1 to 2 viral particles/μL) and specificity (&gt, 97% true negative rate) utilizing reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed. ALERT (Accessible LAMP-Enabled Rapid Test) incorporates the following features: (1) increased shelf-life and ambient temperature storage, compared to liquid reaction mixes, by using wax layers to isolate enzymes from other reagents, (2) improved specificity compared to other LAMP end-point reporting methods, by using sequence-specific QUASR (quenching of unincorporated amplification signal reporters), (3) increased sensitivity, compared to methods without purification through use of a magnetic wand to enable pipette-free concentration of sample RNA and cell debris removal, (4) quality control with a nasopharyngeal-specific mRNA target, and (5) co-detection of other respiratory viruses, such as influenza B, by multiplexing QUASR-modified RT-LAMP primer sets. The flexible nature of the ALERT workflow allows easy, at-home and point-of-care testing for individuals and higher-throughput processing for labs and hospitals. With minimal effort, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific primer sets can be swapped out for other targets to repurpose ALERT to detect other viruses, microorganisms, or nucleic acid-based markers.

Chronic inflammation is a hallmark of atherosclerosis and macrophages play a central role in controlling inflammation at all stages of atherosclerosis. In atherosclerosis, macrophages and monocyte-derived macrophages are continuously exposed to cholesterol, oxidized lipids, cell debris, cytokines, and chemokines. Not only do these stimuli induce a specific macrophage phenotype, but they also interact extensively, leading to macrophage heterogeneity in atherosclerotic plaques. Herein, we review the diverse phenotypes of macrophages, the mechanisms underlying macrophage activation, and the contributions of macrophages to atherosclerosis in this context. We also summarize recent studies on foamy macrophages and monocyte-derived macrophages in plaque during disease progression. We provide a comprehensive overview of transcriptional, epigenetic, and metabolic reprogramming of macrophages and discuss the emerging concepts of targeting cytokines and macrophages to modulate atherosclerosis.

The geometry (size and shape) of gaps is a key determinant in controlling gap closure during wound healing. However, conventional methods for creating gaps result in un‐defined geometries and poorly characterized conditions (cell death factors and cell debris), which can influence the gap closure process. To overcome these limitations, a novel method to create well‐defined geometrical gaps is developed. First, smooth muscle cells (SMCs) are seeded in variously shaped micro‐containers made out of hyaluronic acid hydrogels. Cell proliferation and cell tension induce fibrous collagen production by SMCs predominantly around the edges of the micro‐containers. Upon removal of SMCs, the selectively deposited collagen results in micro‐containers with cell‐adhesive regions along the edges and walls. Fibroblasts are seeded in these micro‐containers, and upon attaching and spreading, they naturally form gaps with different geometries. The rapid proliferation of fibroblasts from the edge results in filling and closure of the gaps. It is demonstrated that gap closure rate as well as closure mechanism is strongly influenced by geometrical features, which points to an important role for cellular tension and cell proliferation in gap closure.

Analyzing persisters at the single-cell level is crucial to properly define their phenotypic traits. However, single-cell analyses are challenging due to the rare and temporary nature of persister cells, thus requiring their rapid and efficient enrichment in a culture. Existing methods to isolate persisters from a bacterial population show important shortcomings, including contamination with susceptible cells and/or cell debris, which complicate subsequent microscopic analyses. We here describe a protocol to enrich persisters in a culture using β-lactam-induced filamentation followed by size separation. This protocol minimizes the amount of cell debris in the final sample, facilitating single-cell studies of persister cells.

Transverse vaginal septum is one of the variants of Mullerian duct anomaly, caused as a result of defective fusion or recanalisation of vaginal and Mullerian organs. At an early age, it commonly presents as primary amenorrhea along with cyclical abdominal pain while later on usually it presents as dyspareunia and infertility. Our 22-year-old patient presented with secondary amenorrhea. It is very unusual for a transverse vaginal septum to cause secondary amenorrhea. MRI and clinical examination raised the suspicion of transverse vaginal septum causing secondary amenorrhea. She attained regular menstrual cycle after septum excision. The proposed theory behind it is obliteration of microperforated transverse vaginal septum because of menstrual blood and cell debris. Thus, a rare possibility of transverse vaginal septum should also be considered as a differential diagnosis of secondary amenorrhea.

Recent pandemic outbreak of the corona-virus disease 2019 (COVID-19) has raised widespread concerns about the importance of the bioaerosols. They are atmospheric aerosol particles of biological origins, mainly including bacteria, fungi, viruses, pollen, and cell debris. Bioaerosols can exert a substantial impact on ecosystems, climate change, air quality, and public health. Here, we review several relevant topics on bioaerosols, including sampling and detection techniques, characterization, effects on health and air quality, and control methods. However, very few studies have focused on the source apportionment and transport of bioaerosols. The knowledge of the sources and transport pathways of bioaerosols is essential for a comprehensive understanding of the role microorganisms play in the atmosphere and control the spread of epidemic diseases associated with them. Therefore, this review comprehensively summarizes the up to date progress on the source characteristics, source identification, and diffusion and transport process of bioaerosols. We intercompare three types of diffusion and transport models, with a special emphasis on a widely used mathematical model. This review also highlights the main factors affecting the source emission and transport process, such as biogeographic regions, land-use types, and environmental factors. Finally, this review outlines future perspectives on bioaerosols.

Human milk (HM) is considered the gold standard for infant nutrition. HM contains macro- and micronutrients, as well as a range of bioactive compounds (hormones, growth factors, cell debris, etc.). The analysis of the complex and dynamic composition of HM has been a permanent challenge for researchers. The use of novel, cutting-edge techniques involving different metabolomics platforms has permitted to expand knowledge on the variable composition of HM. This review aims to present the state-of-the-art in untargeted metabolomic studies of HM, with emphasis on sampling, extraction and analysis steps. Workflows available from the literature have been critically revised and compared, including a comprehensive assessment of the achievable metabolome coverage. Based on the scientific evidence available, recommendations for future untargeted HM metabolomics studies are included.

Initially dismissed as artifacts during sample preparation or as cell debris in bacterial cultures, outer membrane vesicles (OMVs) are now widely accepted as facsimiles of the outer membrane naturally secreted by Gram-negative bacteria. Within the last decades, several studies focused on OMV biogenesis resulting in different, in part complementary models to explain OMV release. Notably, vesicle formation seems to be an essential process as neither a bacterial species nor a mutant lacking vesicle release has been reported so far. Based on the complex physiological roles discussed for OMVs, it is likely that parallel strategies for their production have evolved to ensure their release in diverse conditions. Although, we are still far from a comprehensive mechanistic understanding of OMV release, several studies shed some insights in the processes driving the liberation of OMVs from the bacterial surface. Within this chapter, we will discuss the observations resulting in the current models for OMV formation including their regulation and relevance for bacterial physiology.