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Background: Lymphoid tissue is the first to react to stress, which manifests as characteristic morphological manifestations, based on which it is possible to study the duration of damage. Aims: Study of the dynamics of changes in the histomorphometric characteristics of the rat thymus during regeneration of the musculocutaneous tissue during mechanical thigh trauma. Material and methods: In total, 84 sexually mature rats were enrolled. These were divided into two groups (experiment and control), which were further classified into four subgroups depending on the time elapsed since the injury (1, 3, 15, and 25 days). Mechanical damage was simulated using a setup that transferred kinetic energy (comparable to a 5.6-mm rifle bullet) to tissues. Subsequently, the animals were removed from the experiment by decapitation; their thymus was removed and microscopically examined. In arbitrary units, the total area of the lobules as well as the area of the cortical and cerebral zones was determined. Results: Our research results indicate different phases of changes in the histomorphometric characteristics of the thymus, which correspond to the course of the wound and can be used in forensic practice to determine the duration of the injury. Conclusions: Mechanical trauma, considered as a stress reaction, causes involutive changes in the thymus and is characterized by changes in the area of the thymus cortex and medulla as well as regular dynamics of changes in the thymic lobules at different stages of wound regeneration.

Bruising of strawberry (Fragaria ×ananassa Duch.) fruit is a common mechanical injury that reduces product value. Wound-induced ethylene may accelerate deterioration or decay, affecting strawberry quality and shelf life. However, bruising susceptibility varies among strawberry cultivars. In this study, cultivars Monterey, Sweet Sensation, Radiance, and two proprietary cultivars (Cultivar A and Cultivar B) from a private breeding program were investigated to evaluate their bruising susceptibility and wound response. Bruising consisted of dropping a 28-g steel ball from 27 cm onto individual fruit; unbruised fruit were the primary control, while fruit exposed to 1 μL·L−1 ethylene were used as a check for ethylene response. All fruit were stored at 20 °C, 90% relative humidity (RH), with respiration and ethylene production measured at 2-hour intervals for 24 hours. Appearance observations were recorded daily until decay onset. Peak respiration rates of 30–40 mL CO2·kg−1·h−1 mostly occurred within 4 hours (‘Cultivar B’) to 6 hours (‘Cultivar A’ and ‘Sweet Sensation’) after bruising, except ‘Monterey’, which peaked at 60 mL CO2·kg−1·h−1 in 2 hours, and ‘Radiance’, which reached 70 mL CO2·kg−1·h−1 in 6 hours. Maximum ethylene production rates after bruising were 0.05 to 0.06 μL·kg−1·h−1 for ‘Monterey’, ‘Cultivar A’, and ‘Cultivar B’, 0.10 μL·kg−1·h−1 for ‘Sweet Sensation’, and 0.20 to 0.37 μL·kg−1·h−1 for ‘Radiance’. ‘Cultivar B’, with the lowest ethylene production, exhibited the lowest overall bruising severity, whereas ‘Radiance’, with the highest ethylene production, exhibited the most severe bruising-induced water soaking, and the other cultivars were intermediate, although ‘Monterey’ bruises were more discolored than those of the other cultivars. ‘Monterey’, ‘Radiance’, and ‘Sweet Sensation’ showed more yellowing and browning of the calyx in response to both bruising and ethylene exposure than ‘Cultivar A’ and ‘Cultivar B’. Except for ‘Cultivar B’, bruising and ethylene exposure increased decay severity.

Winter wheat (Triticum aestivum L.) is used as fresh green winter forage worldwide, and its ability to regenerate after mowing determines whether it can be used for forage production; however, the molecular mechanism of regeneration is poorly understood. This study identified long-chain coding and non-coding RNAs in the wheat cultivar “XN9106,” which is cultivated for forage and grain production separately in winter and summer, and analyzed their function during post-mowing regeneration. The results showed that the degradation of carbohydrate plays an important role in regeneration, as demonstrated by decreased carbohydrate content. The increased gene expression of enzymes including β-amylase, β-fructofuranosidase, sucrose synthase, sucrose-6-phosphate synthase, trehalose-6-phosphate synthase, and trehalose-6-phosphate phosphatase in mowed seedlings suggests regeneration is fueled by degraded carbohydrates that provide energy and carbon skeletons for the Krebs cycle and amino acid synthesis. The decreased auxin content relieved the inhibition of cytokinin synthesis, that controls the transition from cell division to cell expansion and stimulates cell expansion and differentiation during the cell expansion phase, and eventually accelerate post-mowing regeneration of seedlings. Additionally, differentially expressed long-chain non-coding RNAs (lncRNAs) might participate in the regulation of gene expression related to carbohydrate metabolism and hormone signal transduction. This study demonstrated the responses of key mRNAs and lncRNAs during post-mowing regeneration of winter wheat and revealed the importance of carbohydrate and hormone during regeneration, providing valuable information for genetic improvement of forage wheat.

The present work represents a pioneering effort, being the first to analyze genomic and transcriptomic data from Vigna unguiculata (cowpea) kinases. We evaluated the cowpea kinome considering its genome-wide distribution and structural characteristics (at the gene and protein levels), sequence evolution, conservation among Viridiplantae species, and gene expression in three cowpea genotypes under different stress situations, including biotic (injury followed by virus inoculation—CABMV or CPSMV) and abiotic (root dehydration). The structural features of cowpea kinases (VuPKs) indicated that 1,293 bona fide VuPKs covered 20 groups and 118 different families. The RLK-Pelle was the largest group, with 908 members. Insights on the mechanisms of VuPK genomic expansion and conservation among Viridiplantae species indicated dispersed and tandem duplications as major forces for VuPKs’ distribution pattern and high orthology indexes and synteny with other legume species, respectively. Ka/Ks ratios showed that almost all (91%) of the tandem duplication events were under purifying selection. Candidate cis-regulatory elements were associated with different transcription factors (TFs) in the promoter regions of the RLK-Pelle group. C2H2 TFs were closely associated with the promoter regions of almost all scrutinized families for the mentioned group. At the transcriptional level, it was suggested that VuPK up-regulation was stress, genotype, or tissue dependent (or a combination of them). The most prominent families in responding (up-regulation) to all the analyzed stresses were RLK-Pelle_DLSV and CAMK_CAMKL-CHK1. Concerning root dehydration, it was suggested that the up-regulated VuPKs are associated with ABA hormone signaling, auxin hormone transport, and potassium ion metabolism. Additionally, up-regulated VuPKs under root dehydration potentially assist in a critical physiological strategy of the studied cowpea genotype in this assay, with activation of defense mechanisms against biotic stress while responding to root dehydration. This study provides the foundation for further studies on the evolution and molecular function of VuPKs.

Winter wheat (Triticum aestivum L.) is used as fresh green winter forage worldwide, and its ability to regenerate after mowing determines whether it can be used for forage production; however, the molecular mechanism of regeneration is poorly understood. This study identified long-chain coding and non-coding RNAs in the wheat cultivar “XN9106,” which is cultivated for forage and grain production separately in winter and summer, and analyzed their function during post-mowing regeneration. The results showed that the degradation of carbohydrate plays an important role in regeneration, as demonstrated by decreased carbohydrate content. The increased gene expression of enzymes including β-amylase, β-fructofuranosidase, sucrose synthase, sucrose-6-phosphate synthase, trehalose-6-phosphate synthase, and trehalose-6-phosphate phosphatase in mowed seedlings suggests regeneration is fueled by degraded carbohydrates that provide energy and carbon skeletons for the Krebs cycle and amino acid synthesis. The decreased auxin content relieved the inhibition of cytokinin synthesis, that controls the transition from cell division to cell expansion and stimulates cell expansion and differentiation during the cell expansion phase, and eventually accelerate post-mowing regeneration of seedlings. Additionally, differentially expressed long-chain non-coding RNAs (lncRNAs) might participate in the regulation of gene expression related to carbohydrate metabolism and hormone signal transduction. This study demonstrated the responses of key mRNAs and lncRNAs during post-mowing regeneration of winter wheat and revealed the importance of carbohydrate and hormone during regeneration, providing valuable information for genetic improvement of forage wheat.

Objective: We recently initiated microcracks, i.e. micron-scale cracks in the collagen networks of cartilage, using both single low-energy impacts and unconfined, cyclic compressions. We also tracked the propagation of microcracks after cyclic compressions simulating 12,000 walking strides. In this study, we aimed to determine the effect of one or more genipin treatments on: (1) the initiation of microcracks under mechanical impacts and (2) the subsequent propagation of microcracks under cyclic, unconfined compression. We hypothesized that treatments with genipin would improve the resistance of cartilage to microdamage, specifically reducing both the initiation of microcracks under impact loading and the propagation of microcracks under cyclic compression. Design: We tested 49 full-thickness, cylindrical osteochondral specimens. We incorporated one or two doses of genipin in between mechanical treatments, i.e. single low-energy mechanical impacts to initiate microcracks and unconfined, cyclic compressions to propagate microcracks. We also imaged specimens using second harmonic generation confocal microscopy, and analyzed the resulting images to quantify changes in morphologies (length, width, and depth) and orientations of microcracks. Finally, we used separate mixed-regression modeling to evaluate the effects of genipin treatments on mechanically induced microcracks. Results: Specimens treated with genipin presented significantly longer and marginally deeper microcracks after mechanical impacts. Two doses of genipin caused significantly longer and wider microcracks under propagation verses one dose. Conclusions: Our results do not support our hypothesis: unfortunately treatments with genipin, and the resulting mechanisms of cross-linking, do not provide resistance to microdamage, quantified as the initiation and propagation of microcracks.

Summary: Background: Acute trauma on intervertebral discs (IVDs) is thought to be one of the risk factors for IVD degeneration. The pathophysiology of IVD degeneration induced by single high impact mechanical injury is not very well understood. The aim of this study was using a post-traumatic IVD model in a whole organ culture system to analyze the biological and biomechanical consequences of the single high-impact loading event on the cultured IVDs. Methods: Isolated healthy bovine IVDs were loaded with a physiological loading protocol in the control group or with injurious loading (compression at 50% of IVD height) in the one strike loading (OSL) group. After another 1 day (short term) or 8 days (long term) of whole organ culture within a bioreactor, the samples were collected to analyze the cell viability, histological morphology and gene expression. The conditioned medium was collected daily to analyze the release of glycosaminoglycan (GAG) and nitric oxide (NO). Results: The OSL IVD injury group showed signs of early degeneration including reduction of dynamic compressive stiffness, annulus fibrosus (AF) fissures and extracellular matrix degradation. Compared to the control group, the OSL model group showed more severe cell death (P ​

ATP is a (co)transmitter and signaling molecule in the CNS. It acts at a multitude of ligand-gated cationic channels termed P2X to induce rapid depolarization of the cell membrane. Within this receptor-channel family, the P2X7 receptor (R) allows the transmembrane fluxes of Na+, Ca2+, and K+, but also allows the slow permeation of larger organic molecules. This is supposed to cause necrosis by excessive Ca2+ influx, as well as depletion of intracellular ions and metabolites. Cell death may also occur by apoptosis due to the activation of the caspase enzymatic cascade. Because P2X7Rs are localized in the CNS preferentially on microglia, but also at a lower density on neuroglia (astrocytes, oligodendrocytes) the stimulation of this receptor leads to the release of neurodegeneration-inducing bioactive molecules such as pro-inflammatory cytokines, chemokines, proteases, reactive oxygen and nitrogen molecules, and the excitotoxic glutamate/ATP. Various neurodegenerative reactions of the brain/spinal cord following acute harmful events (mechanical CNS damage, ischemia, status epilepticus) or chronic neurodegenerative diseases (neuropathic pain, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, amyotrophic lateral sclerosis) lead to a massive release of ATP via the leaky plasma membrane of neural tissue. This causes cellular damage superimposed on the original consequences of neurodegeneration. Hence, blood-brain-barrier permeable pharmacological antagonists of P2X7Rs with excellent bioavailability are possible therapeutic agents for these diseases. The aim of this review article is to summarize our present state of knowledge on the involvement of P2X7R-mediated events in neurodegenerative illnesses endangering especially the life quality and duration of the aged human population.

Excessive mechanical loading or acute trauma to intervertebral discs (IVDs) is thought to contribute to degeneration and pain. However, the exact mechanisms by which mechanical injury initiates and promotes degeneration remain unclear. This study investigates biochemical changes and extracellular matrix disruption in whole-organ human IVD cultures following acute mechanical injury. Isolated healthy human IVDs were rapidly compressed by 5 % (non-injured) or 30 % (injured) of disc height. 30 % strain consistently cracked cartilage endplates, confirming disc trauma. Three days post-loading, conditioned media were assessed for proteoglycan content and released cytokines. Tissue extracts were assessed for proteoglycan content and for aggrecan integrity. Conditioned media were applied to PC12 cells to evaluate if factors inducing neurite growth were released. Compared to controls, IVD injury caused significant cell death. Injury also caused significantly reduced tissue proteoglycan content with a reciprocal increase of proteoglycan content in culture media. Increased aggrecan fragmentation was observed in injured tissue due to increased matrix metalloproteinase and aggrecanase activity. Injured-IVD conditioned media contained significantly elevated interleukin (IL)-5, IL-6, IL-7, IL-8, MCP-2, GROα, and MIG, and ELISA analysis showed significantly increased nerve growth factor levels compared to non-injured media. Injured-disc media caused significant neurite sprouting in PC12 cells compared to non-injured media. Acute mechanical injury of human IVDs ex vivo initiates release of factors and enzyme activity associated with degeneration and back pain. This work provides direct evidence linking acute trauma, inflammatory factors, neo-innervation and potential degeneration and discogenic pain in vivo. Language: en

The effect of stressing sorghum by leaf stripping on the emergence and growth of Amaranthus hybridus was investigated in 2018 at Henderson Research Station in Mazowe, Zimbabwe. The experiment was set up as a randomised complete block design with three treatments replicated 6 times. A control with A. hybridus only, and another with A. hybridus sprayed with atrazine were maintained for comparison. Stripping 2 leaves significantly (P < 0.001) inhibited A. hybridus height, leaf area, and dry weight compared to no stripping. Increasing leaf stripping intensity from 2 to 4 leaves caused an increase in A. hybridus height, leaf area, and dry weight, possibly because intense leaf stripping reduced the number of sorghum leaves available to capture photosynthetically active radiation required for photosynthesis, subsequently reducing the volume of photoassimilates channelled to sorghum roots for allelochemical production. Leaf excision may have created an additional sink that modified assimilate allocation at the expense of roots.

Studies involving amplified fragment length polymorphism (cDNA-AFLP) have often used polyacrylamide gels with radiolabeled primers in order to establish best primer combinations, to analyze, and to recover transcript-derived fragments. Use of automatic sequencer to establish best primer combinations is convenient, because it saves time, reduces costs and risks of contamination with radioactive material and acrylamide, and allows objective band-matching and more precise evaluation of transcript-derived fragments intensities. This study aimed at examining the gene expression of commercial cultivars of P. guajava subjected to water and mechanical injury stresses, combining analyses by automatic sequencer and fluorescent kits for polyacrylamide gel electrophoresis. Firstly, 64 combinations of EcoRI and MseI primers were tested. Ten combinations with higher number of polymorphic fragments were then selected for transcript-derived fragments recovering and cluster analysis, involving 45 saplings of P. guajava. Two groups were obtained, one composed by the control samplings, and another formed by samplings undergoing stress, with no clear distinction between stress treatments. The results revealed the convenience of using a combination of automatic sequencer and fluorescent kits for polyacrylamide gel electrophoreses to examine gene expression profiles. The Unweighted Pair Group Method with Arithmetic Mean analysis using Euclidean distances points out a similar induced response mechanism of P. guajava undergoing water stress and mechanical injury.

Summary Objectives In vitro mechanical injury of articular cartilage is useful to identify events associated with development of post-traumatic osteoarthritis (OA). To date, many in vitro injury models have used animal cartilage despite the greater clinical relevance of human cartilage. We aimed to characterize a new in vitro injury model using elderly human femoral head cartilage and compare its behavior to that of an existing model with adult bovine humeral head cartilage. Design Mechanical properties of human and bovine cartilage disks were characterized by elastic modulus and hydraulic permeability in radially confined axial compression, and by Young's modulus, Poisson's ratio, and direction-dependent radial strain in unconfined compression. Biochemical composition was assessed in terms of tissue water, solid, and glycosaminoglycan (GAG) contents. Responses to mechanical injury were assessed by observation of macroscopic superficial tissue cracks and histological measurements of cell viability following single injurious ramp loads at 7 or 70%/s strain rate to 3 or 14MPa peak stress. Results Confined compression moduli and Young's moduli were greater in elderly human femoral cartilage vs adult bovine humeral cartilage whereas hydraulic permeability was less. Radial deformations of axially compressed explant disks were more anisotropic (direction-dependent) for the human cartilage. In both cartilage sources, tissue cracking and associated cell death during injurious loading was common for 14MPa peak stress at both strain rates. Conclusion Despite differences in mechanical properties, acute damage induced by injurious loading was similar in both elderly human femoral cartilage and adult bovine humeral cartilage, supporting the clinical relevance of animal-based cartilage injury models. However, inherent structural differences such as cell density may influence subsequent cell-mediated responses to injurious loading and affect the development of OA.

Objective: It has been suggested that chondrocyte death by apoptosis may play a role in the pathogenesis of cartilage destruction in osteoarthritis, but the results of in-vivo and in-vitro investigations have been conflicting. To investigate further the cell death in our in-vitro model for traumatic joint injury, we performed a quantitative analysis by electron microscopy (EM) of cell morphology after injurious compression. For comparison, the TUNEL assay was also performed. Design: Articular cartilage explant disks were harvested from newborn calf femoropatellar groove. The disks were subjected to injurious compression (50% strain at a strain rate of 100%/s), incubated for 3 days, and then fixed for quantitative morphological analysis. Results: By TUNEL, the cell apoptosis rate increased from 7±2% in unloaded controls to 33±6% after injury ( P =0.01; N =8 animals). By EM, the apoptosis rate increased from 5±1% in unloaded controls to 62±10% in injured cartilage ( P =0.02, N =5 animals). Analysis by EM also identified that of the dead cells in injured disks, 97% were apoptotic by morphology. Conclusions: These results confirm a significant increase in cell death after injurious compression and suggest that most cell death observed here was by an apoptotic process.

MicroRNA (miR)-146a is known to be overexpressed in osteoarthritis (OA). However, the role of miR-146a in OA has not yet been fully elucidated. In the present study, we applied mechanical pressure of 10 MPa to human chondrocytes for 60 min in order to investigate the expression of miR-146a and apoptosis following the mechanical pressure injury. Normal human chondrocytes were transfected with an miR-146a mimic or an inhibitor to regulate miR-146a expression. Potential target genes of miR-146a were predicted using bioinformatics. Moreover, luciferase reporter assay confirmed that Smad4 was a direct target of miR-146a. The expression levels of miR-146a, Smad4 and vascular endothelial growth factor (VEGF) were quantified by quantitative reverse transcription PCR and/or western blot analysis. The effects of miR-146a on apoptosis were detected by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry. The results indicated that mechanical pressure affected chondrocyte viability and induced the early apoptosis of chondrocytes. Mechanical pressure injury increased the expression levels of miR-146a and VEGF and decreased the levels of Smad4 in the chondrocytes. In the human chondrocytes, the upregulation of miR-146a induced apoptosis, upregulated VEGF expression and downregulated Smad4 expression. In addition, the knockdown of miR-146a reduced cell apoptosis, upregulated Smad4 expression and downregulated VEGF expression. Smad4 was identified as a direct target of miR-146a by harboring a miR‑146a binding sequence in the 3'-untranslated region (3'-UTR) of its mRNA. Furthermore, the upregulation of VEGF induced by miR‑146a was mediated by Smad4 in the chondrocytes subjected to mechanical pressure injury. These results demonstrated that miR-146a was overexpressed in our chondrocyte model of experimentally induced human mechanical injury, accompanied by the upregulation of VEGF and the downregulation of Smad4 in vitro. Moreover, our data suggest that miR-146a is involved in human chondrocyte apoptosis in response to mechanical injury, and may contribute to the mechanical injury of chondrocytes, as well as to the pathogenesis of OA by increasing the levels of VEGF and damaging the transforming growth factor (TGF)-β signaling pathway through the targeted inhibition of Smad4 in cartilage.

Avaliou-se cinco tipos de embalagens (duas plásticas, duas de papelão ondulado e a caixa K de madeira) quanto à proteção fornecida ao tomate variedade Santa Clara durante seu transporte e manuseio. Os tomates acondicionados em tais embalagens foram comparados com frutos controle, isto é, aqueles que não sofreram qualquer tipo de vibração, impacto ou choque, de acordo com a porcentagem daqueles contendo injúrias mecânicas e segundo também sua intensidade. Os tomates foram armazenados à temperatura ambiente por 7 dias, até atingirem 100% de coloração vermelha, e avaliados quanto à incidência de deterioração, perda de peso e sabor (analiticamente através da relação Brix/acidez titulável e por análise sensorial). Os resultados mostraram que os tomates localizados na parte inferior das caixas foram os mais prejudicados e que as ripas de madeira, ásperas e distantes entre si, assim como os pregos da caixa K, foram responsáveis pelos danos mais significativos. As avaliações apontaram a embalagem de papelão ondulado de menor tamanho como a de maior proteção contra injúrias mecânicas nos tomates. Five different packages (2 made of plastic, 2 of cardboard, and 1 made of wood) were evaluated with respect to the protection of tomatoes (Santa Clara variety) during their transportation and handling. These tomatoes were compared to control samples, as related to fruits with no physical impact or vibration, considering the percentage of those fruits with mechanical injuries and their severity. The fruits were stored at ambient temperature for 7 days until they reached full-color and then evaluated regarding the incidence of decay, weight loss, and flavor by Brix/acidity ratio and sensory analysis. The results have shown that the tomatoes on the bottom of the boxes had worse bruises and that the separated clapboards and the clues of wooden boxes caused the most severe injuries. The results also permitted the conclusion that there were less bruised tomatoes in the smaller cardboard package compared to the plastic and wooden boxes.

Edaravone has been shown to reduce ischemia/reperfusion-induced peripheral nerve injury. However, the therapeutic effect of edaravone on peripheral nerve injury caused by mechanical factors is unknown. In the present study, we established a peripheral nerve injury model by crushing the sciatic nerve using hemostatic forceps, and then administered edaravone 3 mg/kg intraperitoneally. The sciatic functional index and superoxide dismutase activity of the sciatic nerve were increased, and the malondialdehyde level was decreased in animals in the edaravone group compared with those in the model group. Bcl-2 expression was increased, but Bax expression was decreased in anterior horn cells of the L4-6 spinal cord segments. These results indicated that edaravone has a neuroprotective effect following peripheral nerve injury caused by mechanical factors through alleviating free radical damage to cells and inhibiting lipid peroxidation, as well as regulating apoptosis-related protein expression.

Background Articular cartilage injury remains a major challenge in orthopedic surgery. This study aimed to identify differences in gene expression and molecular responses between neonatal and adult articular cartilage during the healing of an injury. Methods An established in vitro model was used to compare the transcriptional response to cartilage injury in neonatal and adult sheep by microarray analysis of gene expression. Total RNA was isolated from tissue samples, linearly amplified, and 15,208 ovine probes were applied to cDNA microarray. Validation for selected genes was obtained by real-time quantitative polymerase chain reaction (RT-qPCR). Results We found 1,075 (11.6%) differentially expressed probe sets in adult injured cartilage relative to normal cartilage. A total of 1,016 (11.0%) probe sets were differentially expressed in neonatal injured cartilage relative to normal cartilage. A total of 1,492 (16.1%) probe sets were differentially expressed in adult normal cartilage relative to neonatal normal cartilage. A total of 1,411 (15.3%) probe sets were differentially expressed in adult injured cartilage relative to neonatal injured cartilage. Significant functional clusters included genes associated with wound healing, articular protection, inflammation, and energy metabolism. Selected genes (PPARG, LDH, TOM, HIF1A, SMAD7, and NF-κB) were also found and validated by RT-qPCR. Conclusions There are significant differences in gene expression between neonatal and adult ovine articular cartilage following acute injury. They are partly due to intrinsic differences in the process of development, and partly to different biological responses to mechanical trauma between neonatal and adult articular cartilage.