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Explaining biodiversity is the central focus in theoretical ecology. A significant obstacle arises from the competitive exclusion principle (CEP), which states that two species competing for the same type of resources cannot coexist at constant population densities, or more generally, the number of consumer species cannot exceed that of resource species at steady states. The conflict between CEP and biodiversity is exemplified by the paradox of the plankton, where a few types of limiting resources support a plethora of plankton species. In this review, we introduce mechanisms proposed over the years for promoting biodiversity in ecosystems, with a special focus on those that alleviate the constraints imposed by the CEP, including mechanisms that challenge the CEP in well‐mixed systems at a steady state or those that circumvent its limitations through contextual differences. [ABSTRACT FROM AUTHOR]

SPOUT1/CENP-32 encodes a putative SPOUT RNA methyltransferase previously identified as a mitotic chromosome associated protein. SPOUT1/CENP-32 depletion leads to centrosome detachment from the spindle poles and chromosome misalignment. Aided by gene matching platforms, here we identify 28 individuals with neurodevelopmental delays from 21 families with bi-allelic variants in SPOUT1/CENP-32 detected by exome/genome sequencing. Zebrafish spout1/cenp-32 mutants show reduction in larval head size with concomitant apoptosis likely associated with altered cell cycle progression. In vivo complementation assays in zebrafish indicate that SPOUT1/CENP-32 missense variants identified in humans are pathogenic. Crystal structure analysis of SPOUT1/CENP-32 reveals that most disease-associated missense variants are located within the catalytic domain. Additionally, SPOUT1/CENP-32 recurrent missense variants show reduced methyltransferase activity in vitro and compromised centrosome tethering to the spindle poles in human cells. Thus, SPOUT1/CENP-32 pathogenic variants cause an autosomal recessive neurodevelopmental disorder: SpADMiSS (SPOUT1 Associated Development delay Microcephaly Seizures Short stature) underpinned by mitotic spindle organization defects and consequent chromosome segregation errors. The RNA methyltransferase activity of SPOUT1/CENP-32 is crucial for accurate mitotic spindle organization. Here, the authors describe a neurodevelopmental disorder caused by bi-allelic pathogenic SPOUT1 variants with reduced activity and compromised function in spindle organization. [ABSTRACT FROM AUTHOR]

Intracellular trafficking supports all cell functions maintaining the exchange of material between membrane-bound organelles and the plasma membrane during endocytosis, cargo sorting, and exocytosis/secretion. Several proteins of the intracellular trafficking machinery are deregulated in diseases, particularly cancer. This complex and deadly disease stays a heavy burden for society, despite years of intense research activity. Here, we give an overview about trafficking proteins and highlight that in addition to their molecular functions, they contribute to the emergence of intracellular organelle landscapes. We review recent evidence of organelle landscape alterations in cancer. We argue that focusing on organelles, which represent the higher-order, cumulative behavior of trafficking regulators, could help to better understand, describe and fight cancer. In particular, we propose adopting a physical framework to describe the organelle landscape, with the goal of identifying the key parameters that are crucial for a stable and non-random organelle organization characteristic of healthy cells. By understanding these parameters, we may gain insights into the mechanisms that lead to a pathological organelle spatial organization, which could help explain the plasticity of cancer cells. [ABSTRACT FROM AUTHOR]

Objective: Epothilone D (EpoD), microtubule (MT) stabilizing agent, demonstrated promising results in the animal models of Alzheimer's disease, Parkinson's disease and schizophrenia. The present study sought to investigate preventive effects of EpoD on altered changes of MT related proteins and endoplasmic reticulum (ER) stress proteins induced by social defeat stress (SDS). Methods: We measured protein expression levels of α-tubulin and its post-translational modifications, MT-associated protein 2, stathmin1 and 2 with their phosphorylated forms, and ER stress markers, 78-kDa glucose-regulated protein (GRP-78) and CCAAT/enhancer binding protein (C/EBP)-homologous protein (CHOP) in the prefrontal cortex (PFC) and hippocampus (HIP) of C57BL/6J strain mice treated with EpoD (2 mg/kg) or its vehicle, dimethylsulfoxide (DMSO), and exposed to SDS. Results: We observed lower levels of acetylated α-tubulin, MAP2, p-STMN (Ser16), and GRP-78 in the PFC of the EpoD-Con group when compared to the DMSO-Con group. On the other hand, in the HIP, there were significantly higher levels of tyrosinated α-tubulin and GRP-78 in the EpoD-Defeat group compared to the DMSO-Defeat group. Furthermore, the level of MAP2 in the HIP was found to be lower in the EpoD-Con group compared to the DMSO-Con group. Conclusion: Our results suggest that EpoD exhibits a dual impact, manifesting both beneficial and detrimental effects on the aberrant changes of MT-related proteins and ER stress proteins induced by SDS, depending on the brain regions. These findings underscore the complexity of EpoD's effects, necessitating further exploration to understand its intricate mechanisms in cellular pathways linked to SDS. [ABSTRACT FROM AUTHOR]

Microtubules are cytoskeletal filaments important for various cellular processes such as intracellular transport, cell division, polarization and migration. Microtubule organization goes hand in hand with cellular function. Motile cells, such as immune cells or fibroblasts, contain microtubule asters attached to the centrosome and the Golgi complex, whereas in many other differentiated cells, microtubules form linear arrays or meshworks anchored at membrane-bound organelles or the cell cortex. Over the past decade, new developments in cell culture, genome editing and microscopy have greatly advanced our understanding of complex microtubule arrays. In this Cell Science ata Glance article and the accompanying poster, we review the diversity of microtubule arrays in interphase animal cells. We describe microtubule network geometries present in various differentiated cells, explore the variety in microtubule-organizing centers responsible for these geometries, and discuss examples of microtubule reorganization in response to functional changes and their interplay with cell motility and tissue development. [ABSTRACT FROM AUTHOR]

Marine plankton include organisms driving multiple ecosystem services (ESs). In this study, we identified ESs provided by planktonic consortia worldwide from the analysis of scientific literature. We also mapped the identified ESs onto forty-nine plankton trophic networks derived from presence–absence data obtained from two coastal surveys in three areas along the coast of the Campania region in the Tyrrhenian Sea (NW Mediterranean). The systematic review evidenced that ESs associated with goods provision and ecosystem regulation were the most studied categories, while cultural ESs were the least considered. The mapping of ESs across the Campania coast revealed significant spatiotemporal variations in plankton-based ESs, which depend on seasonal variability and local environmental conditions. Among the ESs, those connected with the regulation and maintenance of marine ecosystems dominated both temporally and spatially, highlighting the crucial role of plankton in ecosystem stability and resilience. Moreover, although the direct provision of goods and materials was less represented, food provision to higher trophic levels was widespread within each coastal sector, highlighting the crucial role of plankton biodiversity in directly sustaining the whole marine ecosystem and related economic activities such as fisheries and aquacultures. [ABSTRACT FROM AUTHOR]

Nitrogen-fixing microorganisms (diazotrophs) significantly influence marine productivity by reducing nitrogen gas into bioavailable nitrogen. Recently, non-cyanobacterial diazotrophs (NCDs) have been identified as important contributors to marine nitrogen fixation. Among them, Gamma A is one of the best-studied marine NCDs because of its ubiquitous occurrence; however, the factors controlling its distribution remain unknown. In particular, the importance of microzooplankton grazing as a top-down control has not yet been examined. In this study, we investigated the diazotroph community structure using nifH amplicon sequencing and quantified the growth and microzooplankton grazing mortality rate of Gamma A using a combination of dilution experiments and quantitative polymerase chain reaction (PCR) in well-lit waters at the northern edge of the Kuroshio Current off the southern coast of Japan. In the study region, Gamma A was ubiquitous and dominant in the diazotroph communities, whereas cyanobacterial diazotrophs had lower relative abundances. The microzooplankton grazing rate of Gamma A was significantly higher than that of the whole phytoplankton community and was generally balanced with its growth rate, suggesting the efficient transfer of fixed nitrogen by Gamma A to higher trophic levels. Although the growth rates of Gamma A did not show clear responses to nutrient amendments, Gamma A abundance had a significant relationship with nutrient concentration and microzooplankton grazing mortality rate. This suggests that microzooplankton grazing, as well as nutrient concentration, plays a vital role in constraining the Gamma A distribution in the Kuroshio region. Our findings highlight the importance of the further in situ quantification of microzooplankton grazing rates to understand the distribution of diazotrophs and the associated nitrogen transfer into the food web. [ABSTRACT FROM AUTHOR]

Settling aggregates transport organic matter from the ocean surface to the deep sea and seafloor. Though plankton communities impact carbon export, how specific organisms and their interactions affect export efficiency is unknown. Looking at 15 years of eDNA sequences (18S-V4) from settling and sedimented organic matter in the Fram Strait, here we observe that most phylogenetic groups were transferred from pelagic to benthic ecosystems. Chaetoceros socialis, sea-ice diatoms, Radiolaria, and Chaetognatha are critical components of vertical carbon flux to 200 m depth. In contrast, the diatom C. socialis alone is essential for the amount of organic carbon reaching the seafloor. Spatiotemporal changes in community composition show decreasing diatom abundance during warm anomalies, which would reduce the efficiency of a diatom-driven biological carbon pump. Interestingly, several parasites are also tightly associated with carbon flux and show a strong vertical connectivity, suggesting a potential role in sedimentation processes involving their hosts, especially through interactions with resting spores, which could have implications for pelagic-benthic coupling and overall ecosystem functioning. Examining 15 years of data in the Arctic, this study shows that plankton, including diatoms and parasites, play key roles in exporting carbon to the seafloor. Species like Chaetoceros socialis are crucial for carbon flux and pelagic-benthic coupling. [ABSTRACT FROM AUTHOR]

The most severe form of muscular dystrophy (MD), known as Duchenne MD (DMD), remains an incurable disease, hence the ongoing efforts to develop supportive therapies. The dysregulation of autophagy, a degradative yet protective mechanism activated when tissues are under severe and prolonged stress, is critically involved in DMD. Treatments that harness autophagic capacities therefore represent a promising therapeutic approach. Osmolytes are protective organic molecules that regulate osmotic pressure and cellular homeostasis and may support tissue-repairing autophagy. We therefore explored the effects of the osmolyte ectoine in the standard mouse model of DMD, the mdx, focusing on the autophagy-related proteome. Mice were treated with ectoine in their drinking water (150 mg/kg) or through daily intraperitoneal injection (177 mg/kg) until they were 5.5 weeks old. Hind limb muscles were dissected, and samples were prepared for Western blotting for protein quantification and for immunofluorescence for an evaluation of tissue distribution. We report changes in the protein levels of autophagy-related 5 (ATG5), Ser366-phosphorylated sequestosome 1 (SQSTM1), heat shock protein 70 (HSP70), activated microtubule-associated protein 1A/1B-light chain 3 (LC3 II) and mammalian target of rapamycin (mTOR). Most importantly, ectoine significantly improved the balance between LC3 II and SQSTM1 levels in mdx gastrocnemius muscle, and LC3 II immunostaining was most pronounced in muscle fibers of the tibialis anterior from treated mdx. These findings lend support for the further investigation of ectoine as a potential therapeutic intervention for DMD. [ABSTRACT FROM AUTHOR]

Colorectal cancer is one of the most common cancers worldwide and has a high mortality rate. In this study, we investigated the cytotoxic, proapoptotic, and anti-invasive effects of the synthetic indole phytoalexin MB-653. The antiproliferative effect was determined using an MTT assay, showing IC50 values of 5.8 ± 0.3 μmol/L for HCT116 cells and 6.1 ± 2.1 μmol/L for Caco2 cells. Flow cytometry and Western blot analysis were employed to investigate the molecular mechanisms underlying cytotoxicity, proapoptotic action, and anti-invasion effects. The proapoptotic activity was evidenced by the activation of caspases 3 and 7, mitochondrial dysfunction, and an increased number of apoptotic cells, confirmed by annexin V/PI and AO/PI staining. Additionally, MB-653 induces dose-dependent G2/M phase cell cycle arrest, the cause of which could be cyclin B1/CDC2 complex dysfunction and/or a decrease in α-tubulin protein expression. Another important observation was that MB-653 modulated several signalling pathways associated with various cellular activities, including survival, proliferation, tumour invasiveness, metastasis, and epithelial–mesenchymal transition (EMT). We further demonstrated its safety for topical and parenteral application. To sum up, our results indicate the real potential of MB-653 in treating colorectal cancer. [ABSTRACT FROM AUTHOR]

To examine the bio-optical response to coastal upwelling, we measured inherent optical properties (IOPs) and biogeochemical parameters simultaneously off Hainan Island in the northwest part of the South China Sea (SCS) during late summer 2013. Bio-optical relationships between IOPs and phytoplankton were used for calculating vertical profiles of the total chlorophyll a concentration (Chl-a) and the absorption by coloured detrital matter (CDM). These bio-optical properties, which showed distinct horizontal and vertical distributions across the continental shelf, were strongly influenced by upwelling processes, as well as the shelf topography. Phytoplankton biomass and CDM absorption in surface waters showed much higher values along the coast, with their spatial distributions related to topographic variability. Vertical distributions of phytoplankton were characterised by a subsurface chlorophyll maximum (SCM) layer. The strongest SCM (Chl-a = 4.22 mg m−3) was observed at 24 m depth in coastal waters near the northeast cape of Hainan Island. The depth of the SCM varied between 16 and 60 m at different stations, appearing to coincide with the isotherm of 22 °C. The SCM depth was inversely correlated with the magnitude of the SCM. Different shapes of Chl-a profiles were observed, which suggested that the vertical distributions of phytoplankton biomass were driven by different environmental factors. Elevated concentrations of CDM were mainly observed near the bottom, which suggest that the benthic nepheloid layer may be an important source of detrital material. The relationship between the absorption coefficient of CDM at 443 nm, aCDM(443), and Chl-a exhibited distinct differences between waters in upper ocean and in bottom layers, with the threshold depth being modulated by shelf topography. Our results highlight the utility of bio-optical observations with high resolution for better understanding the coupling between physical forcing and biogeochemical variability. [ABSTRACT FROM AUTHOR]

The Lim Kinase (LIMK) family of serine/threonine kinases is comprised of LIMK1 and LIMK2, which are central regulators of cytoskeletal dynamics via their well-characterized roles in promoting actin polymerization and destabilizing the cellular microtubular network. The LIMKs have been demonstrated to modulate several fundamental physiological processes, including cell cycle progression, cell motility and migration, and cell differentiation. These processes play important roles in maintaining cardiovascular health. However, LIMK activity in healthy and pathological states of the cardiovascular system is poorly characterized. This review highlights the cellular and molecular mechanisms involved in LIMK activation and inactivation, examining its roles in the pathophysiology of vascular and cardiac diseases such as hypertension, aneurysm, atrial fibrillation, and valvular heart disease. It addresses the LIMKs' involvement in processes that support cardiovascular health, including vasculogenesis, angiogenesis, and endothelial mechanotransduction. The review also features how LIMK activity participates in endothelial cell, vascular smooth muscle cell, and cardiomyocyte physiology and its implications in pathological states. A few recent preclinical studies demonstrate the therapeutic potential of LIMK inhibition. We conclude by proposing that future research should focus on the potential clinical relevance of LIMK inhibitors as therapeutic agents to reduce the burden of cardiovascular disease and improve patient outcomes. Schematic diagram illustrating the contribution of LIM kinases to the pathophysiology of cardiovascular diseases. [ABSTRACT FROM AUTHOR]

The lipid content of nine dinoflagellates was analyzed using flow cytometry to compare lipid levels. Additionally, the correlation between lipid content, cell size, and carbon content in dinoflagellates was evaluated using BODIPY 505/515 staining. The flow cytometry side scatter (SSC) effectively represented relative cell size, showing a linear relationship with the equivalent spherical diameter (ESD). Larger cells exhibited higher relative lipid content; however, lipid accumulation was influenced by nutritional modes and habitats, with mixorophic and benthic species displaying higher lipid content than heterotrophic species. A comparison of fluorescent dyes revealed that Nile Red overestimated lipid content, suggesting overlap with chlorophyll autofluorescence. Flow cytometry analysis with BODIPY 505/515 demonstrated a linear correlation with the sulfo-phospho-vanillin (SPV) method, enabling determination of actual lipid content using FL1 fluorescence and the slope value. As the carbon content increased, the lipid content initially increased rapidly but plateaued at higher carbon levels, indicating saturation. These findings suggest that relative fluorescence via flow cytometry provides an effective means to estimate the lipid content and carbon content as a function of cell size. [ABSTRACT FROM AUTHOR]

In the Nicotiana tabacum flower development study, we identified SCI1 (Stigma/style Cell-cycle Inhibitor 1), a regulator of cell proliferation. SCI1 interacts with NtCDKG;2 (N. tabacum Cyclin-Dependent Kinase G;2), a homolog of human CDK11, which is responsible for RanGTP-dependent microtubule stabilization, regulating spindle assembly rate. In a Y2H screening of a cDNA library using NtCDKG;2 as bait, a RanBP1 (Ran-Binding Protein 1) was revealed as its interaction partner. RanBP1 is an essential regulatory protein of the RanGTPase system, contributing to the formation of the Ran gradient, which modulates different important cellular processes. RanBP1 is crucial in the nuclear import/export machinery during interphase and spindle checkpoint formation during cell division. These processes are well studied in animals, but very little is known about them in plants. We confirmed NtCDKG;2 and NtRanBP1 interaction by pairwise Y2H and characterized the localization of both proteins during plant cell division. We demonstrated the presence of NtRanBP1 in the cytoplasm during interphase and its nuclear arrest at mitosis onset. Meanwhile, we showed that NtCDKG;2 is localized in the mitotic spindle during cell division, indicating an analogous function to the human CDK11. We propose that the phosphorylation of the nuclear export signal at RanBP1 by NtCDKG;2 may be responsible for the reported nuclear arrest. [ABSTRACT FROM AUTHOR]

Mitochondrial Rho-GTPase 1 (MIRO1) is an outer mitochondrial membrane protein which regulates mitochondrial transport and mitophagy in mitosis. In present study, we reported the crucial roles of MIRO1 in mammalian oocyte meiosis and its potential relationship with aging. We found that MIRO1 expressed in mouse and porcine oocytes, and its expression decreased in aged mice. MIRO1 deficiency caused the failure of meiotic resumption and polar body extrusion in both mouse and porcine oocytes, which could be rescued by exogenous MIRO1 supplementation. Mass spectrometry data indicated that MIRO1 associated with several cytoskeleton and cell cycle-related proteins, and MIRO1 regulated motor protein Dynein for microtubule-organizing centers (MTOCs) dynamics at germinal vesicle (GV) stage, which determined meiotic resumption. Furthermore, we found that MIRO1 regulated Aurora A and kinesin family member 11 (KIF11) for meiotic spindle assembly in oocytes. Besides, MIRO1 associated with several mitochondria-related proteins dynamic-related protein 1 (DRP1), Parkin and lysosomal-associated membrane protein 2 (LAMP2) for mitochondrial dynamics and mitophagy during oocyte meiosis. Taken together, our results suggested that MIRO1 played pivotal roles in meiotic resumption, spindle assembly and mitochondrial function in mouse and porcine oocytes, and its insufficiency might contribute to the oocyte maturation defects during aging., Competing Interests: Compliance and ethics. The authors declare that they have no conflict of Interest., (© 2025. Science China Press.)

Background/aims: Endoscopic sphincterotomy (EST) is a standard treatment for common bile duct (CBD) stones. Endoscopic sphincterotomy combined with endoscopic papillary large balloon dilation (EST-EPLBD) is an effective treatment for difficult CBD stones. This study aims to evaluate and compare the effectiveness and adverse effects of EST-EPLBD and EST in treating large CBD stones., Methods: We retrospectively analyzed the data of 85 patients with large CBD stones who underwent either EST or EST-EPLBD, resulting in successful CBD stone extraction from January 2018 to June 2022. Propensity score inverse weighting was performed to reduce the possible bias in baseline characteristics between the two treatment groups. The success rate of complete stone removal in the first session, mechanical lithotripsy use, and adverse events were analyzed by multivariable risk regression analysis., Results: The rate of complete stone removal in one session of the EST-EPLBD group was higher than that of the EST group at 28.78% (95% confidence interval [CI] 4.43, 50.1; p = 0.003). Mechanical lithotripsy use was decreased in the EST-EPLBD group by 25.81% (95% CI 42.33,9.28; p = 0.002). However, the incidence of adverse events is comparable., Conclusion: EST-EPLBD may be utilized in the treatment of CBD stones that exceed a diameter of 10 mm. The EST-EPLBD increased the rate of complete stone removal in a single session and reduced the need for mechanical lithotripsy. Conversely, the incidence rate of adverse events is similar., (© 2024 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

Elongator is a tRNA-modifying complex that regulates protein translation. Recently, a moonlighting function of Elongator has been identified in regulating the polarization of the microtubule cytoskeleton during asymmetric cell division. Elongator induces symmetry breaking of the anaphase midzone by selectively stabilizing microtubules on one side of the spindle, contributing to the downstream polarized segregation of cell-fate determinants, and therefore to cell fate determination. Here, we investigate how Elongator controls microtubule dynamics. Elongator binds both to the tip of microtubules and to free GTP-tubulin heterodimers using two different subcomplexes, Elp123 and Elp456, respectively. We show that these activities must be coupled for Elongator to decrease the tubulin critical concentration for microtubule elongation. As a consequence, Elongator increases the growth speed and decreases the catastrophe rate of microtubules. Surprisingly, the Elp456 subcomplex binds to tubulin tails and has strong selectivity towards polyglutamylated tubulin. Hence, microtubules assembled by Elongator become selectively enriched with polyglutamylated tubulin, as observed in vitro, in mouse and Drosophila cell lines, as well as in vivo in Drosophila Sensory Organ Precursor cells. Therefore, Elongator rewrites the tubulin code of growing microtubules, placing it at the core of cytoskeletal dynamics and polarization during asymmetric cell division., Competing Interests: Disclosure and competing interests statement. The authors declare no competing interests., (© 2025. The Author(s).)

Nucleoside triphosphate (NTP)-dependent protein assemblies such as microtubules and actin filaments have inspired the development of diverse chemically fueled molecular machines and active materials but their functional sophistication has yet to be matched by design. Given this challenge, we asked whether it is possible to transform a natural adenosine 5'-triphosphate (ATP)-dependent enzyme into a dissipative self-assembling system, thereby altering the structural and functional mode in which chemical energy is used. Here we report that FtsH (filamentous temperature-sensitive protease H), a hexameric ATPase involved in membrane protein degradation, can be readily engineered to form one-dimensional helical nanotubes. FtsH nanotubes require constant energy input to maintain their integrity and degrade over time with the concomitant hydrolysis of ATP, analogous to natural NTP-dependent cytoskeletal assemblies. Yet, in contrast to natural dissipative systems, ATP hydrolysis is catalyzed by free FtsH protomers and FtsH nanotubes serve to conserve ATP, leading to transient assemblies whose lifetimes can be tuned from days to minutes through the inclusion of external ATPases in solution., Competing Interests: Competing interests: G.M. is an employee and T.P.J.K. is the founder of Wavebreak Therapeutics. The work reported here was not influenced by this connection. The other authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Fuelled chemical systems have considerable functional potential that remains largely unexplored. Here we report an approach to transient amide bond formation and use it to harness chemical energy and convert it to mechanical motion by integrating dissipative self-assembly and the Marangoni effect in a source-sink system. Droplets are formed through dissipative self-assembly following the reaction of octylamine with 2,3-dimethylmaleic anhydride. The resulting amides are hydrolytically labile, making the droplets transient, which enables them to act as a source of octylamine. A sink for octylamine was created by placing a drop of oleic acid at the air-water interface. This source-sink system sets up a gradient in surface tension, which gives rise to a macroscopic Marangoni flow that can transport the droplets in solution with tunable speed. Carbodiimides can fuel this motion by converting diacid waste back to anhydride. This study shows how fuelling at the molecular level can, via assembly at the supramolecular level, lead to liquid flow at the macroscopic level., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

Biological cells sample their surrounding microenvironments using nanoscale force sensors on the cell surfaces. These surface-based force and stress sensors generate physical and chemical responses inside the cell. The inherently well-connected cytoskeleton and its physical contacts with the force elements on the nuclear membrane lead these physicochemical responses to cascade all the way inside the cell nucleus, physically altering the nuclear state. These physical alterations of the cell nucleus, through yet-unknown complex steps, elicit physical and functional responses from the chromatin in the form of altered gene expression profiles. This mechanism of force/stress sensing by the cell and then its nuclear response has been shown to play a vital role in maintaining robust cellular homeostasis, controlling gene expression profiles during developmental phases as well as cell differentiation. In the last few years, there has been appreciable progress toward the identification of the molecular players responsible for force sensing. However, the actual sensing mechanism of cell surface-bound force sensors and more importantly cascading of the signals, both physical (via cytosolic force sensing elements such as microtubule and actin framework) as well as chemical (cascade of biochemical signaling from cell surface to nuclear surface and further to the chromatin), inside the cell is poorly understood. In this chapter, we present a review of the currently known molecular players in cellular as well as nuclear force sensing repertoire and their possible mechanistic aspects. We also introduce various biophysical concepts and review some frequently used techniques that are used to describe the force/stress sensing and response of a cell. We hope that this will help in asking clearer questions and designing pointed experiments for better understanding of the force-dependent design principles of the cell surface, nuclear surface, and gene expression., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

The latest high-resolution 3D live-cell imaging technology, lattice light-sheet microscopy (LLSM), has successfully tracked the dynamics of microtubule growth throughout the entire mitotic spindle with unparalleled precision. By using green fluorescent protein-labeled end-binding protein 1 (EB1-GFP) as a marker for growing microtubule ends, LLSM has generated an extensive collection of multidimensional datasets mapping the positions and trajectories of these growing microtubule ends. Processing this data requires statistical analysis in three-dimensional space. This chapter describes the spatial statistical methods developed for this purpose, illustrated with practical examples. Finally, we discuss future prospects for analyzing complex, large-scale image data., (© 2025. The Author(s).)

Over the last decade, considerable progress has been made in unraveling RNA virus diversity. This has contributed to our understanding of the evolution of these viruses, which include emerging zoonotic human pathogens. Current success has been greatly facilitated by the development of next-generation sequencing platforms instrumental for meta-transcriptomic studies. However, due to the rapid evolution of RNA viruses, there are numerous "blind spots" waiting to be explored; one of those is the RNA virome of unicellular eukaryotes. Here, we present the pipeline, which has been successfully used to characterize various types of RNA viruses, including Leishbuviridae (Bunyaviricetes, Hareavirales) in the parasitic flagellates of the family Trypanosomatidae. The pipeline relies on axenic in vitro cell culture and double-stranded RNA enrichment, followed by direct RNA-sequencing. A detailed procedure description starting from the initial total RNA preparation to the final assembly of the viral segments is provided., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)