Regeneration Sites Research Articles

Post-Fire Restoration and Deadwood Management: Microsite Dynamics and Their Impact on Natural Regeneration

After large and severe wildfires, the establishment of tree regeneration, particularly for species without specific fire-adaptive traits, can be challenging. Within harsh environments, the presence of favorable microsites, as those provided by deadwood, enhancing microclimatic conditions, is crucial to the re-establishment of forest cover and thus to foster recovery dynamics. Active restoration strategies can have an impact on these dynamics, altering or hindering them. The main hypothesis of this study is that manipulating deadwood in terms of quantity and spatial arrangement can result in differences in natural regeneration density and composition. Post-disturbance regeneration dynamics and the role played by deadwood over time in the creation of safe sites for seedling establishment were investigated in an area affected by a high-severity wildfire that underwent different post-fire restoration treatments along a gradient of increasing deadwood manipulation, spanning from salvage logging to non-intervention. Two inventories were performed 5 and 11 years after the fire. Ground cover proportion was significantly different among treatments, with lower values of lying deadwood in salvaged sites. A higher probability of regeneration establishment close to deadwood was found in both surveys, confirming the facilitating role of deadwood on post-fire forest regeneration. Microsite dynamics resulting from deadwood facilitation were highlighted, with establishment probability and anisotropic relationships between deadwood elements and seedlings changing over time, as recovery processes slowly improved environmental conditions. In dry mountain areas affected by stand-replacing wildfires, by removing deadwood, salvage logging reduces the number of safe sites for regeneration, further impairing the ecosystem recovery. Passive management should be the ecologically preferred management strategy in these conditions, although intermediate interventions (e.g., felling without delimbing, leaving deadwood on the ground) could be effective alternatives, accelerating snag fall dynamics and immediately increasing favorable microsite availability.

Dose-response analysis after administration of a human platelet-derived exosome product on neurite outgrowth in vitro.

Modulating the nerve's local microenvironment using exosomes is proposed to enhance nerve regeneration. This study aimed to determine the optimal dose of purified exosome product (PEP) required to exert maximal neurite extension. An in vitro dorsal root ganglion (DRG) neurite outgrowth assay was used to evaluate the effect of treatment with (i) 5% PEP, (ii) 10% PEP, (iii) 15% PEP, or (iv) 20% PEP on neurite extension (N = 9/group), compared to untreated controls. After 72 h, neurite extension was measured to quantify nerve regeneration. Live cell imaging was used to visualize neurite outgrowth during incubation. Treatment with 5% PEP resulted in the longest neurite extension and was superior to the untreated DRG (p = 0.003). Treatment with 10% PEP, 15% PEP, and 20% PEP was found to be comparable to controls (p = 0.12, p = 0.06, and p = 0.41, respectively) and each other. Live cell imaging suggested that PEP migrated towards the DRG neural regeneration site, compared to the persistent homogenous distribution of PEP in culture media alone. 5% PEP was found to be the optimal concentration for nerve regeneration based on this in vitro dose-response analysis.

The Role of Extracellular Matrix (ECM) Adhesion Motifs in Functionalised Hydrogels.

To create functional tissue engineering scaffolds, biomaterials should mimic the native extracellular matrix of the tissue to be regenerated. Simultaneously, the survival and functionality of stem cells should also be enhanced to promote tissue organisation and repair. Hydrogels, but in particular, peptide hydrogels, are an emerging class of biocompatible scaffolds which act as promising self-assembling biomaterials for tissue engineering and regenerative therapies, ranging from articular cartilage regeneration at joint defects, to regenerative spinal cord injury following trauma. To enhance hydrogel biocompatibility, it has become imperative to consider the native microenvironment of the site for regeneration, where the use of functionalised hydrogels with extracellular matrix adhesion motifs has become a novel, emerging theme. In this review, we will introduce hydrogels in the context of tissue engineering, provide insight into the complexity of the extracellular matrix, investigate specific adhesion motifs that have been used to generate functionalised hydrogels and outline their potential applications in a regenerative medicine setting. It is anticipated that by conducting this review, we will provide greater insight into functionalised hydrogels, which may help translate their use towards therapeutic roles.

Fine-scale genetic structure of Magnolia iltisiana in forest regeneration sites with a history of selective logging

Background: Selective logging is a frequent practice in the Tropical Montane Cloud Forest which can impact forest regeneration and the genetic makeup of successive generations of trees. The spatial clustering of genetically related individuals, fine-scale genetic structure (FSGS), can develop from the reduction of gene dispersal and the decrease in the number of reproductive individuals at the local scale due to selective logging.
 Questions: In regeneration sites with a history of selective logging, does FSGS differ from a site without such a history? Is FSGS stronger in seedlings and saplings relative to juveniles and adults? Is genetic diversity similar among life stages and sites?
 Studied species: Magnolia iltisiana an endemic tree.
 Study site and dates: Sierra de Manantlan Biosphere Reserve, Jalisco, Mexico.
 2020.
 Methods: We evaluated genetic diversity, genetic structure, and FSGS across four life stages (seedlings, saplings, juveniles, and adults) by genotyping 211 individuals with seven nuclear microsatellite loci in two regeneration and one conserved site.
 Results: We found statistically significant FSGS in the two regeneration sites only for seedlings and saplings, while no evidence of FSGS was detected in the conserved site. No differences in genetic diversity estimates and structure were found among life stages.
 Conclusions: Our study does not suggest an effect of selective logging on genetic diversity on the contrasted conditions and an FSGS pattern only in the earlier stages of the regeneration sites in M. iltisiana.

Trap catch data are poor predictors of damage caused by pine weevil (Hylobius abietis) to conifer seedlings

Damage to planted conifer seedlings caused by the pine weevil Hylobius abietis (L.) is a severe and persistent threat to successful forest regeneration in Europe. Various countermeasures are available, which vary in effectiveness, costs and environmental impact, but none are ideal for all situations. Therefore, there is strong interest in robust assessments of damage risks, as they would enable more cost-effective and environmentally friendly forest pest management. It has been suggested that numbers of adult pine weevils caught in host-odour baited traps placed in regeneration sites may be valuable in such risk assessments. However, published studies provide at most weak support for the hypothesis that trap catch data provide adequate predictions of damage. Therefore, we conducted a two-year field study, designed to determine the relationship between weevil trap catch and subsequent damage at 10 regeneration sites in central Sweden. Site factors that might influence pine weevil feeding on seedlings were recorded and used as explanatory variables in the analysis. Stoniness was the only site factor identified as having a significant effect; damage mainly increased with increases in stoniness. No significant correlation was detected between damage to planted conifer seedlings and numbers of pine weevils trapped in the same locations. We suggest that this lack of correlation between weevil numbers and damage is due to planted seedlings only constituting a minor part of the weeviĺs food intake and considerable between-site variation in availability of food sources other than seedlings. Therefore, assessment of pine weevil numbers appears unlikely to be useful for predicting damage risk at specific regeneration sites.

Looking for a magic island in the sea of agriculture: factors affecting forest patch attractiveness for wild boar in human-dominated landscape

Landscape fragmentation carries the risk of habitat loss for many animal species. This may include wild boars, which are one of the main culprits of conflicts with humans, while isolated patches of forest (forest islands) scattered in the agricultural landscape may affect its ecology and the pressure on the crops. This study aimed to understand the importance of small forest islands located in a human-dominated agricultural landscape in the ecology of wild boar. It explores what features, both of the interior and exterior of the islands, influence their use. The study area covered approximately 220,000 ha of northern Poland, where wild boar use of 200 islands distributed across the agricultural landscape was investigated. For this purpose, the method of counting pellet groups was applied on transects of a total length of 51.6 km. In order to compare the use of islands with compact forest complexes, 84 km of transects were made in the latter, divided into developmental stages and forest type. Among the features of the interior of the islands, increasing cover conditions and the main species of trees (pine, spruce and alder) had a positive influence on the wild boar use. The utilization also depended on the presence of human settlements, other islands, and the type of agricultural crop around the island. When comparing island use to compact forest complexes, islands were used more often than forest regeneration sites and mature stands, but less often than pre-thickets. For the 25% most frequently used by wild boar, islands were more attractive than virtually all stand development stages in compact forest complexes. Due to the cover conditions provided, the occurrence of islands in the agricultural space enables wild boars to use areas more distant from compact forest complexes. This may cause the spread of damage caused by wild boars in the fields, but on the other hand it may reduce the pressure on agricultural areas in the immediate vicinity of compact forest complexes. Intensification of hunting pressure in the vicinity of islands with specific characteristics e.g. good cover conditions, considerable distance from human settlements) will increase the effectiveness of wild boar population control in the agricultural landscape and will have a limiting effect on the damage caused by this species in the fields.

A WO3–TiO2 nanorod/CaCO3 photocatalyst with degradation-regeneration double sites for NO2-inhibited and durable photocatalytic NO

As a promising cleaner technology for nitric oxide degradation, photocatalysis has attracted extensive attention, while the main limitations of photocatalytic nitric oxide are that the toxic NO2 is produced easily and the photocatalytic durability was inferior due to the accumulation of photocatalytic products. In this paper, a WO3–TiO2 nanorod/CaCO3 (TCC) insulating heterojunction photocatalyst with degradation-regeneration double sites was prepared by simple grinding and calcining. The effects of CaCO3 loading on the morphology, microstructure and composition of TCC photocatalyst were investigated by SEM, TEM, XRD, FT-IR and XPS etc. Also, TCC exhibits NO2-inhibited and durable characteristics for NO degradation. DFT calculation, the detection of active radicals by EPR, capture test and the NO degradation pathway characterized by in-situ FT-IR spectra showed that the electron-rich region formed and the existence of regeneration sites are the main reasons for promoting the NO2-inhibited and durable NO degradation. Furthermore, the mechanism of NO2-inhibited and durable NO degradation by TCC was revealed. Finally, TCC superamphiphobic photocatalytic coating was prepared, which still exhibits similar NO2-inhibited and durable characteristics for NO degradation to TCC photocatalyst. It may provide new application value and development prospects in the field of photocatalytic NO.

Developing and Applying an Urban Resilience Index for the Evaluation of Declining Areas: A Case Study of South Korea's Urban Regeneration Sites.

This study attempts to identify the direction of urban regeneration projects in declining areas by using the concept of urban resilience to cope with climate change and disaster. To this end, urban resilience was classified into a Green Resilient Infrastructure (GRI) and an Interactive Safety System (ISS), through a review of previous studies, and categorized into vulnerability, adaptability, and transformability. A total of 12 detailed indicators were derived and indexed using Euclidean distance. Using the indicators, three Korean urban regeneration targets, in Daegu, Mokpo, and Seosan, were selected to evaluate resilience before and after the urban regeneration plan. Consequently, the postplanning resilience index improved in all three target sites, compared to before the regeneration plan. Additionally, previously the regeneration plan showed lower index values in comparison to places not designated as urban regeneration areas. These results suggest that urban resilience needs to be considered in future urban regeneration projects, and that resilience indicators can be used as a means to set the direction of urban regeneration projects. To improve the overall resilience of a region, these indices can help local government establish a reference point for urban resilience in its region.

An Antisense Oligonucleotide-Loaded Blood-Brain Barrier Penetrable Nanoparticle Mediating Recruitment of Endogenous Neural Stem Cells for the Treatment of Parkinson's Disease.

Parkinson's disease (PD) is a neurodegenerative disease characterized by the death of dopaminergic (DA) neurons and currently cannot be cured. One selected antisense oligonucleotide (ASO) is reported to be effective for the treatment of PD. However, ASO is usually intrathecally administered by lumbar puncture into the cerebral spinal fluid, through which the risks of highly invasive neurosurgery are the major concerns. In this study, ZAAM, an ASO-loaded, aptamer Apt 19S-conjugated, neural stem cell membrane (NSCM)-coated nanoparticle (NP), was developed for the targeted treatment of PD. NSCM facilitated the blood-brain barrier (BBB) penetration of NPs, and both NSCM and Apt 19S promoted the recruitment of the neural stem cells (NSCs) toward the PD site for DA neuron regeneration. The behavioral tests demonstrated that ZAAM highly improved the efficacy of ASO on PD by the targeted delivery of ASO and the recruitment of NSCs. This work is a heuristic report of (1) nonchemoattractant induced endogenous NSC recruitment, (2) NSCM-coated nanoparticles for the treatment of neurodegenerative diseases, and (3) systemic delivery of ASO for the treatment of PD. These findings provide insights into the development of biomimetic BBB penetrable drug carriers for precise diagnosis and therapy of central nervous system diseases.

Evaluation of instrumentation and pedicle screw design for posterior lumbar fixation: A pre-clinical in vivo/ex vivo ovine model.

Stabilization procedures of the lumbar spine are routinely performed for various conditions, such as spondylolisthesis and scoliosis. Spine surgery has become even more common, with the incidence rates increasing ~30% between 2004 and 2015. Various solutions to increase the success of lumbar stabilization procedures have been proposed, ranging from the device's geometrical configuration to bone quality enhancement via grafting and, recently, through modified drilling instrumentation. Conventional (manual) instrumentation renders the excavated bony fragments ineffective, whereas the "additive" osseodensification rotary drilling compacts the bone fragments into the osteotomy walls, creating nucleating sites for regeneration. This study aimed to compare both manual versus rotary Osseodensification (OD) instrumentation as well as two different pedicle screw thread designs in a controlled split animal model in posterior lumbar stabilization to determine the feasibility and potential advantages of each variable with respect to mechanical stability and histomorphology. A total of 164 single thread (82 per thread configuration), pedicle screws (4.5× 35 mm) were used for the study. Each animal received eight pedicles (four per thread design) screws, which were placed in the lumbar spine of 21 adult sheep. One side of the lumbar spine underwent rotary osseodensification instrumentation, while the contralateral underwent conventional, hand, instrumentation. The animals were euthanized after 6- and 24-weeks of healing, and the vertebrae were removed for biomechanical and histomorphometric analyses. Pullout strength and histologic analysis were performed on all harvested samples. The rotary instrumentation yielded statistically (p= 0.026) greater pullout strength (1060.6N ± 181) relative to hand instrumentation (769.3N ± 181) at the 24-week healing time point. Histomorphometric analysis exhibited significantly higher degrees of bone to implant contact for the rotary instrumentation only at the early healing time point (6 weeks), whereas bone area fraction occupancy was statistically higher for rotary instrumentation at both healing times. The levels of soft tissue infiltration were lower for pedicle screws placed in osteotomies prepared using OD instrumentation relative to hand instrumentation, independent of healing time. The rotary instrumentation yielded enhanced mechanical and histologic results relative to the conventional hand instrumentation in this lumbar spine stabilization model.

Dual-functionalized Pickering HIPE templated poly(ɛ-caprolactone) scaffold for maxillofacial implants

High internal phase emulsion (HIPE) templated poly (ɛ-caprolactone) (PCL) scaffolds have gained widespread attention for large-sized bone defects due to its tuneable 3D architecture and ease of fabricating crosslinked PCL (cPCL) scaffolds. However, extremely high stabilizer (surfactant or nanoparticle) concentration and negligence of microenvironment for regeneration sites like alveolar bones have restrained industrial acceptance of these scaffolds. Herein, we demonstrated the fabrication of nanocomposite cPCL scaffolds within Pickering HIPE templates stabilized using modified silica nanoparticles (mSiNP) concentrations as low as 0.1 to 1.0 wt%. Using an unconventional approach, the mSiNP Pickering stabilizer was added in dispersed phase, contradicting Bancroft’s rule. The colloidal stability was attained due to faster drifting of mSiNP towards the interface when it was dispersed in silicone oil. Scaffolds with tuneable properties were fabricated by controlling the mSiNP concentration and ϕd. Further, cPCL scaffolds were functionalized using clove oil (CO) to improve their efficiency in eradicating S. aureus and E. coli by disrupting their cellular integrity. Additionally, formation of biofilm on the surface of the scaffolds was successfully inhibited by the incorporation of CO. CO-functionalized scaffolds demonstrated excellent cytocompatibility towards MG-63 cells allowing their successful adhesion and proliferation on the surface of the scaffolds.

A bio-orthogonally functionalized chitosan scaffold with esterase-activatable release for nerve regeneration

Developing nerve conduits with biological cues is a promising approach for repairing peripheral nerve injuries. Although most biological cues incorporated into conduits generally exert their biological functions at the surface, they could not be released into the on-demand regeneration sites under physiological conditions. Herein, we firstly report a bio-orthogonally functionalized chitosan scaffold with esterase-activatable release for peripheral nerve regeneration. In this study, biological cues are not only selectively conjugated into nerve conduits by bio-orthogonal reaction, but also precisely released in on-demand regeneration sites via esterase-activatable cleavage for peripheral nerve repair. Moreover, this nerve scaffold with esterase-activatable release could promote Schwann cells proliferation. In a rat sciatic nerve defect model, the bio-orthogonally functionalized scaffold with esterase-activatable release significantly increased sciatic nerve function recovery and improved target muscles weight. This strategy of incorporating esterase-activatable bioactive cues into peripheral nerve conduits offers great potential in preclinical studies.

Damage caused by an exceptionally warm and dry early summer on newly planted Norway spruce container seedlings in Nordic boreal forests

With climate change, drought and warm growth conditions in the spring and early summer are predicted to become more common in Nordic boreal zones in the future. Such conditions occurred in Finland in June and July 2021, offering a good opportunity to study the field performance of seedlings and which factors, including the weather, regeneration site, and operational or seedling level factors, affected the field performance of the seedlings. In the survey, 65 regeneration sites planted with Norway spruce (Picea abies (L.) Karst.) container seedlings from May to July 2021 were randomly selected from different parts of South and Central Finland (60–64°N), and several variables were assessed in sample plots from these sites in September–October 2021. On average, 6 % of the seedlings died, and 26 % were damaged during the first growing season. In most cases, the damage was caused by drought. The most important factors affecting drought risk were the planting period and packaging and storage methods of the seedlings: in freezer-stored seedlings, the probability of drought damage was <0.20 for all planting periods, except for July; when the seedlings overwintered outdoors and were delivered to the forest in open trays, the risk of damage was <0.25 when they had been planted in May and early June, but in late June, it was > 0.60. When the seedlings had overwintered outdoors and were then packed in closed packaging, the risk was >0.20, regardless of the planting period. A low previous-year height of the seedlings (especially in the outdoor stored seedlings), precipitation a week before planting, and a low average temperature two weeks after planting reduced the risk of drought damage. When the average air temperature was below 15 °C (i.e., during the May plantings), the shade of the nearby stand slightly reduced the drought risk, but in higher temperatures, shade did not affect it. The effect of the pre-planting precipitation was stronger in coarse-textured and peat soils than in medium-coarse soils. The risk of drought damage was lowest when the freezer-stored seedlings were planted in good quality mineral soil mounds, deep enough in medium-rich (MT type) and medium-coarse soils. It is impossible to completely prevent drought damage in newly planted Norway spruce seedlings, but by optimizing operational choices and measures, the development of full density fast-growing young Norway spruce forests can also be ensured in a warming climate.

Characteristics of Red Spruce (Picea rubens Sarg.) Encroachment at Two Central Appalachian Heathland Study Areas

During the late 19th and early 20th centuries, intensive land use nearly eliminated red spruce (Picea rubens Sarg.) throughout portions of West Virginia (WV). Red spruce has been slow to regenerate on mountaintop heathland barrens surrounding Canaan Valley, West Virginia (WV), and little is known about the nature of encroachment. Using field surveys, geospatial data, and statistical modelling, the objectives were to 1) characterize and compare red spruce encroachment at two upland heath study areas in West Virginia (Bear Rocks and Cabin Mountain), 2) characterize percent cover of major ground cover types associated with red spruce regeneration sites in order to elucidate biotic interactions, and 3) model the biophysical correlates of red spruce encroachment using geospatial data and statistical modelling. Red spruce count was similar at both study areas, but there were substantially more seedlings and saplings at Cabin Mountain. Modelling revealed a positive relationship between red spruce count and rock cover and a negative relationship between red spruce and stand distance.

DAR 16-II Primes Endothelial Cells for Angiogenesis Improving Bone Ingrowth in 3D-Printed BCP Scaffolds and Regeneration of Critically Sized Bone Defects.

Bone is a highly vascularized tissue and relies on the angiogenesis and response of cells in the immediate environmental niche at the defect site for regeneration. Hence, the ability to control angiogenesis and cellular responses during osteogenesis has important implications in tissue-engineered strategies. Self-assembling ionic-complementary peptides have received much interest as they mimic the natural extracellular matrix. Three-dimensional (3D)-printed biphasic calcium phosphate (BCP) scaffolds coated with self-assembling DAR 16-II peptide provide a support template with the ability to recruit and enhance the adhesion of cells. In vitro studies demonstrated prompt the adhesion of both human umbilical vein endothelial cells (HUVEC) and human mesenchymal stem cells (hMSC), favoring endothelial cell activation toward an angiogenic phenotype. The SEM-EDS and protein micro bicinchoninic acid (BCA) assays demonstrated the efficacy of the coating. Whole proteomic analysis of DAR 16-II-treated HUVECs demonstrated the upregulation of proteins involved in cell adhesion (HABP2), migration (AMOTL1), cytoskeletal re-arrangement (SHC1, TMOD2), immuno-modulation (AMBP, MIF), and morphogenesis (COL4A1). In vivo studies using DAR-16-II-coated scaffolds provided an architectural template, promoting cell colonization, osteogenesis, and angiogenesis. In conclusion, DAR 16-II acts as a proactive angiogenic factor when adsorbed onto BCP scaffolds and provides a simple and effective functionalization step to facilitate the translation of tailored 3D-printed BCP scaffolds for clinical applications.

Survival and Growth of Olive Tree (Olea africana) Seedlings Under Open Field Conditions is Enhanced in Hydrogel Amended Soils Media, Teso Subcounty, Kenya

The rapid decline of timber from natural forests against increasing domestic demand for wood products has caused a steady increase in timber prices in Kenya. Recently, research and development institutions have recognized the ecological and economic value of indigenous tree species and therefore have emphasized the importance of their future production on private smallholder farms rather than in large plantations. This situation has been worsened by climate changes such as the frequent high intensity of prolonged droughts and the nature of soils with poor water retention capacity. As a result, unplanted forest regeneration sites have increased hence a need for re-afforestation of these sites to meet the target for the plantation industry. Therefore, there is a need for mass production of Olea africana seedlings for reforestation programs. One such approach is the application of hydrogels or synthetic polymer organic combinations capable of improving the water and nutrient retention of the soil that can support seedling growth. This study sought to investigate the effects of hydrogel application on selected growth characteristics of O. africana. Seeds were germinated and transferred into two sets of tubes of the same size arranged in a complete randomized block design (CRBD) experiment. The sets were subjected to different levels of hydrogel concentration. Under (0gm as control) and 7gm, 14gm, and 21gm) concentration levels of hydrogel mixed with soil in open field conditions. The height and shoot collar diameter of germinated seedlings were measured every two weeks for two months. The survival of O. africana seedlings decreased with time for all treatments irrespective of the addition of hydrogels with the highest (85%) survival of seedlings on soils mixed with 21gm of hydrogel compared to 0gm hydrogel that recorded (15% survival) eight weeks after transplanting. Application of different levels of hydrogel on soils had a significant influence on the survival of transplanted O. africana seedlings (p˂0.05). Generally, the height of O. africana seedlings increased with time for all types of treatments (0, 7, 14 and 21gm of hydrogel); however, the rate of growth of seedlings was highest (0.667cm /week) on soils treated with 21gm of hydrogel and least (0.33cm/week) on 0gm hydrogel. The shoot collar diameter of O. africana seedlings increased with time for all types of treatments (0, 7, 14 and 21gm of hydrogel); however, the average shoot collar diameter of O. africana seedlings was highest (0.088mm/week) on soils inoculated with 21gm hydrogel and least (0.066mm/week) on soils not inoculated with hydrogel. These results imply that a higher dose of hydrogel significantly enhances the survival and growth characteristics (height and shoot collar diameter) of transplanted seedlings by improving the water-holding capacity of soil and making it available for plant uptake during dry conditions. Hydrogels can therefore be used to amend soils for a sustainable mass regeneration of tropical timber species for reforestation programs, increased forest cover and restoration of biodiversity in Kenya

Strongylopus grayii tadpole blastema extract exerts cytotoxic effects on embryonal rhabdomyosarcoma cells

Amphibians have regenerative capacity and are resistant to developing cancer. This suggests that the developing blastema, located at the tissue regeneration site, may secrete anti-cancer factors. Here, we investigate the anti-cancer potential of tadpole tail blastema extracts (TAD) from the stream frog, Strongylopus grayii, in embryonal rhabdomyosarcoma (ERMS) cells. ERMS originates in skeletal muscle tissue and is a common pediatric soft tissue sarcoma. We show using MTT assays that TAD inhibited ERMS cell viability in a concentration-dependent manner, and phase contrast/fluorescent microscopy revealed that it induced morphological markers of senescence and apoptosis. Western blotting showed that this was associated with DNA damage (γH2AX) and activation of the p38/MAPK stress signaling pathway as well as molecular markers of senescence (p16INK4a), apoptosis (cleaved PARP), and inhibition of cell cycle promoters (cyclin A, CDK2, and cyclin B1). Furthermore, proteomics followed by gene ontology analyses showed that TAD treatment inhibited known tumor promoters and proteins required for cancer cell survival. Lastly, using the LINCS drug perturbation library, we show that there is an overlap between the proteomics signature induced by TAD and common anti-cancer drugs. Taken together, this study provides novel evidence that TAD exhibits cytotoxicity in ERMS cells.

Management and landscape of shelterbelts contribute to butterfly and flowering plant diversity in northern Japan

AbstractA considerably large number of shelterbelts were established in Japan to increase crop yields in agricultural landscapes. Shelterbelt landscapes have multiple functions; they not only help to control wind damage but also to conserve biodiversity, including forest and grassland species. However, previous studies on biodiversity conservation in shelterbelts, and the target taxa were limited since this aspect of shelterbelts has not received much attention in Japan. This study examines the effects of shelterbelts and their regeneration on the diversity of butterflies and flowering plants in the Tokachi region of Hokkaido, Japan. Using a transect method, we compared environmental factors, butterfly, and flowering plant diversity in four habitats: inside forest, forest edge, regeneration sites, and semi‐natural grasslands which include shelterbelts and surrounding landscapes. There were significant differences in photosynthetically active radiation (PAR) among habitats, with higher PAR corresponding to higher numbers of butterfly species and individuals, and flowering plant species and flowers. Nonmetric multidimensional scaling results also showed that the species composition of flowering plants and butterflies differed among habitats. In addition, indicator value analysis showed that semi‐natural grasslands and regeneration sites were characterized by endangered butterflies and their host plants. Our results indicated that shelterbelt regeneration and the surrounding environment (e.g., semi‐natural grasslands) contributed to the diversity of butterflies and flowering plants, including endangered species. Future studies evaluating the diversity of various taxa would provide a more comprehensive understanding of the effects of shelterbelts on biodiversity conservation.

Increased Arthrodesis Rates in Charcot Neuroarthropathy Utilizing Distal Tibial Distraction Osteogenesis Principles.

Charcot neuroarthropathy of the hindfoot and ankle poses substantial challenges due to deformity, segmental bone loss, chronic infection, and difficulty with bracing. Hindfoot or ankle arthrodesis is often employed at high rates of complications and nonunion. This study reports 15 consecutive patients with Charcot neuroarthropathy who underwent tibiotalocalcaneal or tibiocalcaneal fusion with simultaneous distal tibial distraction osteogenesis with a mean follow-up period of 20.2 ± 5.66 months. Arthrodesis rate was 93.3% (14 patients) with mean time to fusion of 4.75 ± 3.4 months. One hypertrophic nonunion occurred at the arthrodesis site. Complete consolidation of 4 cortices was achieved at the distraction site in 93.3% of patients (14 patients) with a mean duration to consolidation of 9.8 ± 3.3 months. One patient experienced hypertrophic nonunion at the regeneration site. The authors report a technique to enhance arthrodesis rates in Charcot neuroarthropathy by combining distal tibial distraction osteogenesis with simultaneous tibiotalocalcaneal or tibiocalcaneal arthrodesis for hindfoot fusion and salvage. Distraction osteogenesis supports enhanced vascularity to the arthrodesis site.Level of Clinical Evidence: Level 4.

X-ray Micro-Computed Tomography Characterization of Autologous Teeth Particle used in Postextraction Sites for Bone Regeneration. An Experimental Study in Dogs

Objectives: The objective of this study was to develop a new computed method to characterize and measure the bone density measured in Hounsfield units (HU) of particulate tooth grafts, evaluated by micro-computed tomography (micro-CT) at 2 months of healing. Materials and Methods: Thirty-two dog teeth were crushed with a smart dentin grinder, later implanted in postextraction sites of 4 beagle dogs. Twenty-four cores were taken after 2 months and analyzed by micro-CT (Albira). The methodology used was based on a descriptive statistic of the bone density values measured in HU obtained from the creation of volumes of interest (VOIs) and predefined three-dimensional iso-contours from the images obtained after performing micro-CT of the biopsies of the crushing tooth. Results: The micro-CT allows established the characteristics of the biomaterials by studying the HU. The most predominant type of bone was type D3 density (400–800 HU). There was a light presence of bone-type density D2 and D1 in 2 of the regions studied. Conclusions: Micro-CT could be considered a technique of great value in the characterization of biomaterials based on the HU, after implantation in an in vivo model. The distribution of D1 and D2 particles were located around the bottom and middle part of the alveoli and the D3 and D4 bone particles were in the hole core. Therefore, the method proposed in this study is useful to determine the density of the tooth granulate (dentin grinder) and any other biomaterial.