Applied Biosciences

Bone regeneration and repair are complex processes with the potential of added complications, like delayed repair, fracture non-union, and post-surgical infections. These conditions remain a challenge globally, pressurizing the economy and patients suffering from these conditions. Applications of nanotechnology (NBT) in the field of medicine have provided a medium for several approaches to support these global challenges. Tissue engineering is one such field that has been on the rise in the last three decades through the utilization of NBT for addressing the challenges related to bone regeneration. First, NBT enables the formation of scaffolds at the nanoscale needed for bone tissue engineering (BTE) using natural and synthetic polymers, as well as with minerals and metals. Then, it aids the development of the nano-formulation strategized to deliver antimicrobial drugs and/or growth factors through various ways to enhance bone repair through the scaffold. Third, NBT facilitates the use of specialized nanoparticles to image and track cellular events in vitro as well as in vivo. This review is an effort to bring together the current knowledge in the field of BTE and present the scope of ever-evolving NBT, a contribution towards precision medicine. Full article

The chaperone system (CS) is emerging as a key multistage participant in carcinogenesis. The CS chief components are the molecular chaperones (some of which are named heat shock proteins or Hsp), which are typically cytoprotective but if abnormal in structure, location, or quantity, can become etiopathogenic and cause diseases, known as chaperonopathies, including some cancers. For example, abnormal Hsp90 expression is associated with tumorigenesis and poor prognosis. Hsp90 is positioned at the center of several key oncogenic pathways by stabilizing and activating oncogenic kinases responsible for driving cell proliferation and survival. Consequently, inhibition of Hsp90 is being investigated as a possible anti-cancer strategy and some results are encouraging. However, the 5-year survival rate for patients suffering from salivary gland carcinomas is still unsatisfactory. Because of the rarity of these malignancies, they may have been overlooked and understudied and, thus, novel therapies (e.g., inhibition of CS components like Hsp90 and others) are urgently needed. In this review, we also summarize the histopathological quantitative patterns and the intra- and extra-cellular location characteristics of Hsp90 in tumors of salivary glands, pointing to their potential for differential diagnosis, prognostication, and patient monitoring. Full article

In apple (Malus domestica), the level and timing of crop load have a major impact on the final fruit size and can also play a role in optimising internal fruit quality. Ideal crop loads vary with cultivar, but very few cultivars have recommended crop load targets that consider the effect of crop load on both return bloom and fruit quality. To address this issue, studies examining a range of crop loads and thinning times were undertaken on several apple cultivars. Return bloom and multiple fruit quality parameters were examined. The results of these studies demonstrate positive effects for early thinning, not only on fruit size but also on firmness and soluble solids content. Early-thinned fruit showed higher sugar levels than late-thinned fruit. Previously undemonstrated positive relationships between fruit sugar content and weight and between fruit firmness and weight in both ‘Fuji’ and ‘Delicious’, as well as between fruit sugar content and fruit firmness in ‘Delicious’, indicate that early thinning is a valuable tool in improving fruit quality. The current target crop load recommendations of 4–6 fruit cm⁻² trunk cross-sectional area (TCSA) for ‘Fuji’ and 2–4 fruit cm⁻² TCSA for ‘Delicious’ are confirmed by this study. New recommendations are proposed for the other cultivars in this study taking into account the impact of crop load on both fruit quality and return bloom. Both ‘Pink Lady’ and ‘Gala’ can support crop loads of up to eight fruit cm⁻² TCSA without impacting return bloom, but fruit quality is compromised; hence, lower targets in the range of 4–6 fruit cm⁻² TCSA are recommended. Large fruit size and good return bloom can be maintained in ‘Jonagold’ at crop loads of eight fruit cm⁻² TCSA, while crop loads of four fruit cm⁻² TCSA are suggested for ‘Braeburn’ to sustain regular bearing and good fruit size. Full article

Single Nucleotide Polymorphisms (SNPs) are variations that occur at single nucleotides in the genome and are present at an appreciable level in a population. SNPs can be linked to phenotypes of interest, for example diseases, recent adaptations, or species hybridization. They can also be used to study phylogeny and evolutionary history. Technologies that rapidly identify and catalog the presence of SNPs in a DNA sample are known as SNP genotyping panels, and they continue to undergo rapid development. Such methods have great utility across the agricultural sciences in diverse areas such as plant and animal breeding, pathogen and pesticide resistance identification, outbreak tracing, and hybridization detection. Here, we provide an overview of 14 different SNP genotyping technologies and weigh some of the pros and cons associated with each platform. This review is not comprehensive or technical, nor does it aim to be. Rather, the objective is to provide an introduction to the landscape of genotyping technologies for researchers who do not have experience with these methods. Three classes of SNP genotyping methods are Polymerase Chain Reaction (PCR)-based (nine different methods), microarray-based (one method), and Next-Generation Sequencing (NGS)-based (four different methods). We discuss how each genotyping class is suited for different niches; PCR-based has a low SNP count and high sample number, microarray-based has a very high SNP count and a moderate sample number, and Next-Generation Sequencing-based has a moderate SNP count and moderate number of samples. Included are basics about how the methods function and example use cases of each method. Additionally, we introduce and discuss the potential for the MinION sequencer in SNP genotyping. For each technology, we provide insights into cost, equipment needs, labor costs, experimental complexity, data output complexity, and accessibility. These considerations address the feasibility of deploying the technologies in an agricultural science environment. Full article

Anterior tooth (ANT) contacts induce a short-latency reflex inhibition of the human jaw-closing muscles. The jaw is a rigid class 1 lever for pinpoint targeting muscle force into a single bite point, the pivoting food particle. Seesaw reflex movements around the food particle fulcrum multiply the food-crushing force. Unpredictable jolts of reaction force caused by food crushing are subjected to the rostral ANT and caudally to the two articulate ends of the jaw triangle. The compression/distraction strains of food crushing must be monitored and inhibited by withdrawal reflexes. The mesencephalic ganglion (Vmes), neural myelin sheath, and muscle stretch receptors evolved subsequently to the advent of jaws to improve the velocity of proprioceptive and withdrawal reflexes. In mammalians, the spindles of the taut motor units, stretched by the food fulcrum, send excitatory monosynaptic feedback for the efferent neurons of the respective ipsilateral muscle units via the Vmes. In the Vmes, the spindle-input-mediating afferent neurons are coupled with another source of afferent feedback, which is also excitatory, from the back tooth (BAT) mechanoreceptors. The two sources of excitatory pulses are summated and targeted for the efferent neurons to boost the stretched and taut motor units. Likewise, the afferent feedback from the ANT mechanoreceptors is also coupled in the Vmes with concomitant feedback from spindles. The ANT output, however, is inhibitory to negate the excitatory feedback from the stretched jaw muscle units. The inhibitory feed from the anterior teeth temporarily blocks the excitatory potential of the masticatory motor efferent neurons to protect the anterior teeth and jaw joints from inadvertent strains. The inhibitory inputs from the anterior teeth alternate with the excitatory inputs from the BAT to determine which jaw-closing muscle units are activated or inhibited at any given instant of food crushing. The Vmes exists in all jawed vertebrates, and its evolution was probably motivated by demands for the control of bite force. The monosynaptic unilateral food-crushing excitatory and inhibitory reflexes (UFCRs) override the coexisting bilaterally executed feed for the jaw muscles from the central nervous system. The hypothesis proposed in this study is that the Vmes-mediated UFCRs combine neural inputs from tooth contacts with concomitant feedback from the muscle stretch receptors for the control of the mammalian food-crushing bite force. Full article

It has been previously reported that time-varying EMFs and LEDs have the potential to modulate cellular activity and cell viability. It has also been shown that cellular activity and state can be inferred by measuring the biophoton emission derived from these same cells. To identify if the brief application (15 min) of an LED (635 nm at 3 klx) or EMF (1–3 uT) could influence cell growth and subsequent biophoton emission characteristics, B16-BL6 cells were grown to confluence and exposed to a time-varying, frequency-modulated EMF, LED, or both. Before and after EMF and LED exposure, photon emission measurements were taken for 1 min at a 50 Hz sampling rate. Following the exposure and photon emission measurements, cell viability was assessed via the use of a hemocytometer. The results demonstrated that after only 15 min of exposure to a time-varying EMF, there was a 41.6% reduction in viable cells when compared to sham controls [t(25) = 2.4, p = 0.02]. This effect approached significance in the LED alone condition [p = 0.07] but was completely absent in the condition wherein the LED and EMF were applied simultaneously [p < 0.8]. Additionally, following exposure to only the LED, there was a significant increase in biophoton emission SPD values at 13 Hz from whole cell cultures [t(60) = 2.3, p = 0.021]. This biophoton emission frequency was also strongly correlated with the number of nonviable cells [r = −0.514] in the dish. Taken together, these data point to biophotons emitted from cell cultures at 13 Hz as a potential indicator of the number of nonviable cells in vitro. The summation of data here corroborates previous work demonstrating the efficacy of specific time-varying EMFs as a novel therapeutic for the inhibition of cancer cell growth. It also furthers our assertion that biophoton emission can be used as a novel detection tool for cell activity. Full article

An imperative requisite of tissue-engineered scaffolds is to promote host cell regeneration and concomitantly thwart microbial growth. Antibacterial agents are often added to prevent implant-related infections, which, however, aggravates the risk of bacterial resistance. For the first time, we report a fiber-based platform that selectively promotes the growth of mammalian cells and alleviates bacteria by varying fiber size, orientation, and material of polymeric yarns. The interactions of Gram-positive and -negative bacterial species with mammalian mesenchymal stem cells (MSC) were investigated on poly-€-caprolactone (PCL) yarns, polyethylene terephthalate (PET), poly-L-lactic acid (PLLA), and cotton. Various yarn configurations were studied by altering the fiber diameter (from nano- to microscale) and fiber orientations (aligned, twisted, and random) of PCL yarns. PCL nanofibrous yarn decreased the adhesion of S. aureus and E. coli, with a 2.7-fold and 1.5-fold reduction, respectively, compared to PCL microfibrous yarn. Among different fiber orientations, nanoaligned fibers resulted in an 8-fold and 30-fold reduction of S. aureus and E. coli adhesion compared to random fibers. Moreover, aligned orientation was superior in retarding the S. aureus adhesion by 14-fold compared to nanotwisted fibers. Our data demonstrate that polymeric yarns comprising fibers with nanoscale features and aligned orientation promote mammalian cell adhesion and spreading and concomitantly mitigate bacterial interaction. Moreover, we unveil the wicking of cells through polymeric yarns, facilitating early cell adhesion in fibrous scaffolds. Overall, this study provides insight to engineer scaffolds that couple superior interaction of mammalian cells with high-strength fibrous yarns for regenerative applications devoid of antibacterial agents or other surface modification strategies. Full article

Bacterial contamination of water and food sources is still a major source of diseases. Early detection of potential pathogens is key to prevent their spreading and severe health risks. Here, we describe a fast, low-cost detection assay based on horseradish peroxidase (HRP) conjugated to streptavidin for the direct identification of bacteria. Streptavidin can bind to bacterial cells due to its high affinity for biotin, a natural component of microbial cell surfaces. Upon binding to bacteria, the HRP converts a chromogenic substrate, resulting in a visible color change. In the present study, we evaluated different detection platforms regarding their compatibility with the detection principle. To reduce background signals and increase the sensitivity of HRP-based assays, the binding of HRP to surfaces and biomolecules was intensively investigated. The final assay successfully detected the most relevant bacterial strains in drinking water, such as Escherichia coli, Klebsiella pneumonia, and Enterobacter cloacae. Full article

Dermatoses are essentially caused by infection or free radical aggression, immunoallergic disorders, or can be secondary to general diseases. Management of dermatoses by modern medicine is complex and costly, and the development of alternative treatments is urgent. Opilia amentacea Roxb. is a woody climber plant traditionally used in Burkina Faso for treatment of bad skin diseases. This study was carried out to evaluate the antimicrobial and antioxidant activities of extracts of O. amentacea and to characterize potent fractions. The antimicrobial activity was determined using the disc diffusion and microdilution methods, while antioxidant activity was assessed using the 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) and ferric reducing antioxidant power (FRAP) assays. The content of the plant extracts in polyphenols and flavonoids was also studied. The results revealed several secondary metabolites in the leaves, stems and root bark extracts of the plant, including sterols, triterpenes, and flavonoids and tannins, and a generally high total polyphenol and total flavonoid content. Dichloromethane fractions of leaves (FDFe) and stem barks (FDET) exhibited the best antioxidant activity and were the most active on Gram-positive bacilli. Hexane leaves (FHFe) and hexane root bark (FHER) fractions exhibited the best antifungal activity against Candida tropicalis. High correlation (R² = 0.932) was found between the total flavonoid content of extracts and ferric-reducing antioxidant power. In view of these results, the present study describes O. amentacea as a potential source of antibacterial, antifungal and antioxidant agents and justifies the traditional uses of the plant as an anti-dermatosis plant. Full article

The interest in under-utilized crops as a functional food for animals and humans has been increasing recently with advancing research and the need for crop improvement. Canadian forage crops including alfalfa (Medicago sativa L.) and fenugreek (Trigonella foenum-graecum L.) are marketed in various forms due to their traditionally known health benefits. Sainfoin (Onobrychis viciifolia Scop.) is another forage crop with potential health benefits containing beneficial nutraceuticals. In this study, we assessed selected bioactive phenolic compounds and fatty acids in seeds and seedlings of Canadian-grown alfalfa, sainfoin, and fenugreek. Various phenolic compounds were detected in all three forage crop seeds and seedlings. In general, Sainfoin seeds were high in phenolic compounds relative to that of alfalfa and fenugreek. Chlorogenic acid, epigallo catechin, and gallic acid were at high concentrations at 56.6, 86.8, and 64.7 µg.g⁻¹, respectively, compared to other phenolic compounds in sainfoin seeds. The fatty acids content (%) was significantly affected by the seedling stage and crop type. Some of the bioactive compounds present in seeds were not detected in seedling stages. The comparative bioactive phenolic compounds and fatty acid assessments of these forage legumes could potentially be used as biomarkers for the selection and development of favorable cultivars for animal and human nutrition. In addition, these crops could be used for isolating these bioactive compounds, and thus increasing their agri-food value. Full article

Deregulated transcription is a well-known characteristic of cancer cells, with differentially expressed genes being a common feature of several cancers. Often, deregulated transcription is a consequence of alterations in transcription factors (TFs), which play a crucial role in gene expression and can act as tumour suppressors or proto-oncogenes. In eukaryotic organisms, transcription is carried out by three distinct RNA polymerase complexes: Pol I, Pol II, and Pol III. Pol II, specifically, is responsible for transcribing messenger RNA (mRNA), the protein coding part of the genome, as well as long non-coding RNAs (lncRNAs). While there is considerable research on the impact of specific deregulated transcription factors in cancer development, there is a lack of studies focusing on defects within the RNA polymerase complexes and their subunits. This review aims to shed light in particular on the Pol II complex and highlight the deregulation of its subunits that have a significant impact on tumour development, prognosis, and survival. By providing a comprehensive overview of our current understanding of Pol II subunits in cancer, this review emphasizes the importance of further research in this area. It suggests that exploring these subunits’ deregulations could lead to the identification of valuable biomarkers and potential therapeutic targets, making it a topic of collective interest. Full article

Rapid and accurate pathogen identification is crucial in effectively combating infectious diseases. However, the current diagnostic tools for bacterial infections predominantly rely on century-old culture-based methods. Furthermore, recent research highlights the significance of host–microbe interactions within the host microbiota in influencing the outcome of infection episodes. As our understanding of science and medicine advances, there is a pressing need for innovative diagnostic methods that can identify pathogens and also rapidly and accurately profile the microbiome landscape in human samples. In clinical settings, such diagnostic tools will become a powerful predictive instrument in directing the diagnosis and prognosis of infectious diseases by providing comprehensive insights into the patient’s microbiota. Here, we explore the potential of long-read sequencing in profiling the microbiome landscape from various human samples in terms of speed and accuracy. Using nanopore sequencers, we generate native DNA sequences from saliva and stool samples rapidly, from which each long-read is basecalled in real-time to provide downstream analyses such as taxonomic classification and antimicrobial resistance through the built-in software (<12 h). Subsequently, we utilize the nanopore sequence data for in-depth analysis of each microbial species in terms of host–microbe interaction types and deep learning-based classification of unidentified reads. We find that the nanopore sequence data encompass complex information regarding the microbiome composition of the host and its microbial communities, and also shed light on the unexplored human mobilome including bacteriophages. In this study, we use two different systems of long-read sequencing to give insights into human microbiome samples in the ‘slow’ and ‘fast’ modes, which raises additional inquiries regarding the precision of this novel technology and the feasibility of extracting native DNA sequences from other human microbiomes. Full article

As schools go digital, the use of tablet computers is increasing. Concerns are raised that the extensive use of tablets and the associated bent-over posture may negatively affect the individual’s health. In order to analyse the possible effects of prolonged tablet use on physical health, a detailed analysis of the posture during tablet use is needed so that appropriate preventive measures can be taken to prevent degenerative changes. Therefore, the aim of this study was to measure and report the posture of 56 students while working with a tablet computer and compare it with an upright posture. Sagittal and frontal images were used for measurements of the subjects’ postures while seated, using the tablet, and in a neutral sitting position looking straight ahead. The body position during tablet use was recorded in two different user configurations: tablet flat on the table and tablet in individual freely chosen user configuration. After appropriate annotation of the data, the following parameters were evaluated in different planes. The craniovertebral angle (CVA), head tilt angle (HTA), and forward shoulder angle (FSA) are measurements that describe the extent to which the head bends forward and downward and how the shoulders are aligned in the sagittal plane. On the other hand, the head shoulder angle (HSA), lateral head tilt angle (LHTA), and trunk flexion angle (TFA) are angles measured in the frontal plane, which indicate the degree of head tilt and trunk bending to the right or left side. The measurement results clearly showed that the use of a tablet had a pronounced effect on the positions and rotations of the participants’ head, neck, and shoulders. This was evident through strong deviations observed in the angles measured between the sitting straight posture and the postures while using the tablet. For example, depending on the body posture class, the mean CVA values were 45.76° for straight sitting posture, 28.25° for holding the tablet individually posture, and 26.04° for the posture adopted while using a tablet placed flat on the table. Full article

Two-P-domain K⁺ (K2p) channels are responsible for maintaining the resting membrane potential. K2p channels have varied expression in healthy tissue, but they also change in cancerous or diseased states. The correlation and causation as regards the alteration of K2p channel expression are still being investigated. The compound doxapram seems to block K2p channels and depolarize cells. Using Drosophila, the increased expression of the ORK1 K2p channel in cardiac and skeletal muscle was investigated. The heart rate in larval Drosophila is very sensitive to pH, and since doxapram blocks a subset of the K2p channels that are known to be acid-sensitive, it was postulated that doxapram would affect heart rate. A pH change from 7.1 to 6.5 increased the rate, while that from 7.1 to 7.5 decreased the rate. An amount of 0.1 mM of doxapram had no effect, but 0.5 of mM depressed Drosophila heart rates within five minutes. Exposure to 5 mM of doxapram immediately decreased the rate. Lipopolysaccharides (LPSs) from Gram-negative bacteria acutely increased the rate. LPSs activate K2p channels in the skeletal muscle of larvae and are blocked by doxapram. LPSs slightly reduce depression in the rate induced by doxapram. The overexpression of K2p channels in the heart and skeletal muscle depressed the heart rate and heightened pH sensitivity. At larval neuromuscular junctions, the overexpression in skeletal muscle increases the frequency of spontaneous quantal events and produces a more negative resting membrane potential. Full article

The build-up of lipofuscin—an age-associated biomarker referred to as hyperfluorescence—is considered a precursor in the progression of geographic atrophy (GA). Prior studies have attempted to classify hyperfluorescent regions to explain varying rates of GA progression. In this study, digital image processing and unsupervised learning were used to (1) completely automate the extraction of hyperfluorescent regions from images, and (2) evaluate prospective patterns and groupings of hyperfluorescent areas associated with varying levels of GA progression. Patterns were determined by clustering methods, such as k-Means, and performance was evaluated using metrics such as the Silhouette Coefficient (SC), the Davies–Bouldin Index (DBI), and the Calinski–Harabasz Index (CHI). Automated extraction of hyperfluorescent regions was carried out using pseudocoloring techniques. The approach revealed three distinct types of hyperfluorescence based on color intensity changes: early-stage hyperfluorescence, intermediate-stage hyperfluorescence, and late-stage hyperfluorescence, with the early and late stages having three additional subclassifications that could explain varying levels of GA progression. The performance metrics for early-stage hyperfluorescence were SC = 0.597, DBI = 0.915, and CHI = 186.989. For late-stage hyperfluorescence, SC = 0.593, DBI = 1.013, and CHI = 217.325. No meaningful subclusters were identified for the intermediate-stage hyperfluorescence, possibly because it is a transitional phase of hyperfluorescence progression. Full article

Medicago sativa (2n = 4x = 32) and M. arborea (2n = 4x = 32) were thought to be reproductively isolated until hybrids (Alborea) were produced by sexual reproduction for the first time in 2003 in Wisconsin. The hybrids were asymmetric, at or near 2n = 4x = 32, and with a predominance of the alfalfa genome. Only M. sativa seed parents with reproductive abnormalities, including unreduced eggs, have produced hybrids; where M. arborea has been used as the seed parent, no hybrids have resulted. Pedigree selection within derivatives of the two original M. sativa seed parents (MB and M8) has been successful in increasing the frequency of hybrids produced. While Alborea individuals more closely resemble M. sativa, a number of M. arborea-specific traits have been observed across different hybrid individuals. These include single-coil flat pods, large seeds, yellow flowers, indeterminate growth, a minimal crown, lodging, frost resistance, and anthracnose resistance. These M. arborea traits have the potential to restructure alfalfa to increase its versatility and utilisation. There is emerging evidence from North and South America and Australia that some Alborea selections have the capacity to complement adapted alfalfa cultivars for yield. Work is continuing to introgress M. arborea traits of value into alfalfa. Full article

Red wine grapes have an important impact on the economy of many regions, both for wine quality and for their richness in phenolic compounds, which have many health benefits. Climate has been changing substantially in the last years, which affects greatly grape polyphenolic composition and wine quality. In this review, we will unveil the importance of climate in grape development, both physically and chemically, the different methodologies used to evaluate grape quality, the interesting new approaches using NIR spectroscopy, and the functional properties of grapes and red wine, due to their high phenolic content. Climate has an impact in the development of phenolic compounds in grapes, namely in the anthocyanins biosynthesis. The phenolic chemical composition changes during maturation, therefore, it is essential to keep on track the accumulation of these key compounds. This information is crucial to help producers choose the best harvest date since specific compounds like polyphenols are responsible for the color, taste, and mouthfeel of wines, which directly affects wine quality. The usage of different methodologies to assess quality parameters in grapes and wine, can be used to provide essential information to create the chemical profile of each variety to develop calibration methods. NIR spectroscopy seems to be a reliable method to be used in vineyards during grape maturation to provide real time information on quality parameters to producers since many reliable calibration models have been developed over time. Full article

The metabolic diversity of soil microbiota embodies diverse functional capabilities that support ecosystem resilience, driving essential biogeochemical processes and facilitating the optimization of sustainable agricultural systems. Integrating cover crops into agricultural systems cultivates a diverse array of metabolic activities among soil microbes, synergistically enhancing ecosystem services and bolstering soil health for sustainable and productive farming practices. In an effort to gain deeper insights and expand our knowledge, we conducted a study examining the effects of cover crops and fertilizer sources, thereby shedding light on their combined impacts on the metabolic activity dynamics of soil microbial communities. In this investigation, we employed a split-plot design with two factors: (a) cover crop with three solo cover crop species—Cereal rye (Secale cereale), wheat (Triticum aestivum), hairy vetch (Vicia villosa), and one mixture of mustard (Brassica rapa) and cereal rye (Secale cereale) (CC-mix), (b) Fertilizer source includes poultry litter, chemical fertilizer, and no-fertilizer treatments. We assessed the metabolic potential of soil microbiota by using carbon substrates utilizing Biolog EcoPlates. The findings revealed that the plots with CC-mix treatment exhibited greater metabolic diversity compared to the other treatments, while among the fertilizer sources, poultry litter demonstrated higher metabolic activity. Furthermore, both treatment factors predominantly metabolized carbohydrates and polymers compared to other carbon substrate categories. The principal component analysis accounted for 46.4% of the variance, collectively represented by PC1 and PC2, emphasizing the substantial contributions of carbohydrates, amino acids, and carboxylic acids to the observed metabolic diversity. Canonical correspondence analysis revealed that pH had positively correlated with microbial functional diversity, whereas total carbon (TC), total nitrogen (TN), and water-stable aggregates (WSA) showed a negative correlation. In conclusion, cover cropping and type of fertilizer source had a notable impact on soil microbial functional diversity, with the cover crop mixture exhibiting a more pronounced influence than the individual cover crop treatments. Full article

Background: Expansion of OKF6/TERT-2 oral epithelial cells in vitro is important for studying the molecular biology of disease and pathology affecting the oral cavity. Keratinocyte serum-free medium (KSFM) is the medium of choice for this cell line. This study compares three media for OKF6/TERT-2 cultures: KSFM, Dulbecco’s Modified Eagle Medium/Nutrient Mixture of Hams F-12 (DMEM/F12), and a composite medium comprised of DMEM/F-12 and KSFM (1:1 v/v), referred to as DFK. The toxicological effects of electronic cigarette liquids (e-liquids) on OKF6/TERT-2 cells cultured in these media were also compared. Methods: Cells were cultured in KSFM, DMEM/F12, or DFK, and cellular morphology, growth, wound healing and the gene expression of mucins and tight junctions were evaluated. Additionally, cytotoxicity was determined after e-liquid exposures. Results: Switching from KSFM to DMEM/F12 or DFK 24 h post-seeding leads to typical cellular morphologies, and these cultures reach confluency faster than those in KSFM. Wound-healing recovery occurred fastest in DFK. Except for claudin-1, there is no difference in expression of the other genes tested. Additionally, e-liquid cytotoxicity appears to be amplified in DFK cultures. Conclusions: DMEM/F12 and DFK are alternative media for OKF6/TERT-2 cell culture to study the molecular biology of disease and pathology, provided cells are initially seeded in KSFM. Full article