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141.
Chromatin investigation in the nucleus using a phasor approach to structured illumination microscopy
Isotta Cainero Elena Cerutti Mario Faretta Gaetano Ivan Dellino Pier Giuseppe Pelicci Paolo Bianchini Giuseppe Vicidomini Alberto Diaspro Luca Lanzanò 《Biophysical journal》2021,120(12):2566-2576
Chromatin in the nucleus is organized in functional sites at variable level of compaction. Structured illumination microscopy (SIM) can be used to generate three-dimensional super-resolution (SR) imaging of chromatin by changing in phase and in orientation a periodic line illumination pattern. The spatial frequency domain is the natural choice to process SIM raw data and to reconstruct an SR image. Using an alternative approach, we demonstrate that the additional spatial information encoded in the knowledge of the position of the illumination pattern can be efficiently decoded using a generalized version of separation of photon by lifetime tuning (SPLIT) that does not require lifetime measurements. In the resulting SPLIT-SIM, the SR image is obtained by isolating a fraction of the intensity corresponding to the center of the diffraction-limited point spread function. This extends the use of the SPLIT approach from stimulated emission depletion microscopy to SIM. The SPLIT-SIM algorithm is based only on phasor analysis and does not require deconvolution. We show that SPLIT-SIM can be used to generate SR images of chromatin organizational motifs with tunable resolution and can be a valuable tool for the imaging of functional sites in the nucleus. 相似文献
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The axolotl, Ambystoma mexicanum, is used extensively for research in developmental biology, particularly for its ability to regenerate and restore lost organs, including in the nervous system, to full functionality. Regeneration in mammals typically depends on the healing process and scar formation with limited replacement of lost tissue. Other organisms, such as spiny mice (Acomys cahirinus), salamanders, and zebrafish, are able to regenerate some damaged body components. Blastema is a tissue that is formed after tissue injury in such organisms and is composed of progenitor cells or dedifferentiated cells that differentiate into various cell types during regeneration. Thus, identifying the molecules responsible for initiation of blastema formation is an important aspect for understanding regeneration. Introns, a major source of noncoding RNAs (ncRNAs), have characteristic sizes in the axolotl, particularly in genes associated with development. These ncRNAs, particularly microRNAs (miRNAs), exhibit dynamic regulation during regeneration. These miRNAs play an essential role in timing and control of gene expression to order and organize processes necessary for blastema creation. Master keys or molecules that underlie the remarkable regenerative abilities of the axolotl remain to be fully explored and exploited. Further and ongoing research on regeneration promises new knowledge that may allow improved repair and renewal of human tissues. 相似文献
146.
Marta Cambra Sergio Madrigal-Mora Isaac Chinchilla Geiner Golfín-Duarte Christopher G. Lowe Mario Espinoza 《Journal of fish biology》2021,99(3):1140-1144
Tiger sharks (Galeocerdo cuvier) play an important ecological role as top predators, yet knowledge of their reproductive ecology is scarce. Here, the authors report the first observation of a potential neonate G. cuvier at Cocos Island, a predator-dominated oceanic island in the Eastern Tropical Pacific (ETP). The individual was detected using baited remote underwater video stations (BRUVS). The cameras also detected female individuals potentially pregnant, suggesting that parturition may take place at or near the island. Nonetheless, it is still unclear if the presence of a single neonate is an isolated event or evidence that the species is using the island for reproduction. 相似文献
147.
Roadways may pose barriers to long-distance migrators such as some ungulates. Highway underpasses mitigate wildlife-vehicle collisions and can be an important management tool for protecting migration corridors. In northern California, 3 underpasses were built on United States Route 395 (Route 395) in Hallelujah Junction Wildlife Area (HJWA) in the 1970s for a migratory mule deer (Odocoileus hemionus) herd that had been negatively affected by highway traffic. To determine whether these underpasses were still reducing mule deer mortalities >40 years after construction, we investigated deer use of the underpasses from 2006–2019 using cameras, global positioning system (GPS) collars, and roadkill records. We used occupancy models, approximations of GPS-collared mule deer movement paths, and roadkill locations to estimate the highway crossing patterns of deer. From camera data, there was higher use of the underpasses by deer during migration (spring [Mar–Jun], fall [Oct–Dec]) than in summer (Jul–Sep), when only resident deer were present. Higher underpass usage occurred in the spring compared to fall migrations. Eleven of 21 GPS-collared migrating mule deer crossed Route 395. We estimated 30% of the crossings (by 7 of the 11 deer) occurred south of the underpasses where deer could easily access the highway because of short (1-m high) and deteriorating highway fencing. Roadkill data confirmed that deer-vehicle collisions were occurring south of the underpasses and at the underpasses. This was likely due to deteriorating infrastructure at the underpasses that allows wildlife access to the highway. Overall, our study indicated that although underpasses can provide safe passage for migratory deer decades (>40 yr) after their construction, deteriorating infrastructure such as fencing and gates can lead to wildlife mortalities on highways near underpasses. © 2021 The Wildlife Society. 相似文献
148.
Mario Rincón-Barrado Sanna Olsson Tamara Villaverde Belén Moncalvillo Lisa Pokorny Alan Forrest Ricarda Riina Isabel Sanmartín 《植物分类学报:英文版》2021,59(5):915-934
Wide-range geographically discontinuous distributions have long intrigued scientists. We explore the role of ecology, geology, and dispersal in the formation of these large-scale disjunctions, using the angiosperm tribe Putorieae (Rubiaceae) as a case study. From DNA sequences of nuclear ITS and six plastid markers, we inferred a phylogeny with 65% of all known Putorieae species. Divergence times, ancestral ranges, and diversification rate shifts were then estimated using Bayesian inference. We further explored species climatic tolerances and performed ancestral niche reconstruction to discriminate among alternative speciation modes, including geographical and ecological vicariance, and ecogeographical, ecological, and dispersal-mediated speciation. As a result, we identified seven major clades in Putorieae, some of which exhibit striking geographical disjunctions, matching the Rand Flora pattern, with sister species in the Canary Islands andeastern and southern Africa. Initial diversification within the tribe occurred in the early Miocene, coincident with a period of climate warming; however, most clades diverged within the last 10 Myr. Aridification and high extinction rates, coupled with ecological vicariance, explain the oldest disjunctions. Adaptation to new environmental conditions, after allopatry, is observed in several clades. Dispersal, either long-distance or via corridors made available by mountain uplift, is behind the most recent disjunctions. Some of these events were followed by ecological speciation and rapid diversification, with species becoming adapted to xeric or increasingly colder continental climates. We show that an integrative approach may help discriminate among speciation modes invoked to explain disjunctions at macroevolutionary time scales, even when extinction has erased the signature of past events. 相似文献
149.
Mario Gimona Maria Felice Brizzi Andre Boon Hwa Choo Massimo Dominici Sean M. Davidson Johannes Grillari Dirk M. Hermann Andrew F. Hill Dominique de Kleijn Ruenn Chai Lai Charles P. Lai Rebecca Lim Marta Monguió-Tortajada Maurizio Muraca Takahiro Ochiya Luis A. Ortiz Wei Seong Toh Yong Weon Yi Sai Kiang Lim 《Cytotherapy》2021,23(5):373-380
Mesenchymal stromal/stem cells (MSCs) have been widely tested against many diseases, with more than 1000 registered clinical trials worldwide. Despite many setbacks, MSCs have been approved for the treatment of graft-versus-host disease and Crohn disease. However, it is increasingly clear that MSCs exert their therapeutic functions in a paracrine manner through the secretion of small extracellular vesicles (sEVs) of 50–200 nm in diameter. Unlike living cells that can persist long-term, sEVs are non-living and non-replicative and have a transient presence in the body. Their small size also renders sEV preparations highly amenable to sterilization by filtration. Together, acellular MSC-sEV preparations are potentially safer and easier to translate into the clinic than cellular MSC products. Nevertheless, there are inherent challenges in the development of MSC-sEV drug products. MSC-sEVs are products of living cells, and living cells are sensitive to changes in the external microenvironment. Consequently, quality control metrics to measure key identity and potency features of MSC-sEV preparations have to be specified during development of MSC-sEV therapeutics. The authors have previously described quantifiable assays to define the identity of MSC-sEVs. Here the authors discuss requirements for prospective potency assays to predict the therapeutic effectiveness of the drug substance in accordance with International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines. Although potency assays should ideally reflect the mechanism of action (MoA), this is challenging because the MoA for the reported efficacy of MSC-sEV preparations against multiple diseases of diverse underlying pathology is likely to be complex and different for each disease and difficult to fully elucidate. Nevertheless, robust potency assays could be developed by identifying the EV attribute most relevant to the intended biological activity in EV-mediated therapy and quantifying the EV attribute. Specifically, the authors highlight challenges and mitigation measures to enhance the manufacture of consistent and reproducibly potent sEV preparations, to identify and select the appropriate EV attribute for potency assays despite a complex “work-in-progress” MoA and to develop assays likely to be compliant with regulatory guidance for assay validation. 相似文献
150.
Anelise Sonza Luciana Sayuri Sanada Luiza Raulino de Oliveira Mario Bernardo-Filho Danúbia da Cunha de S-Caputo Milton Antonio Zaro Matilde Achaval 《Experimental biology and medicine (Maywood, N.J.)》2021,246(10):1210
Whole-body vibration (WBV), which is widely used as a type of exercise, involves the use of vibratory stimuli and it is used for rehabilitation and sports performance programmes. This study aimed to investigate the effect of WBV treatment in a chronic pain model after 10 WBV sessions. An animal model (chronic pain) was applied in 60 male Wistar rats (±180 g, 12 weeks old) and the animals were treated with low intensity exercise (treadmill), WBV (vibrating platform), and a combined treatment involving both. The controls on the platform were set to a frequency of 42 Hz with 2 mm peak-to-peak displacement, g ≈ 7, in a spiral mode. Before and after the vibration exposure, sensitivity was determined. Aβ-fibers-mediated mechanical sensitivity thresholds (touch-pressure) were measured using a pressure meter. C-fibers-mediated thermal perception thresholds (hot pain) were measured with a hot plate. After each session, WBV influenced the discharge of skin touch-pressure receptors, reducing mechanical sensitivity in the WBV groups (P < 0.05). Comparing the conditions “before vs. after”, thermal perception thresholds (hot pain) started to decrease significantly after the third WBV session (P < 0.05). WBV decreases mechanical hyperalgesia after all sessions and thermal sensitivity after the third session with the use of WBV. 相似文献