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The sex determination system of largemouth bass (Micropterus salmoides, LMB) is XX/XY; however, the underlying molecular mechanisms involved in early sex differentiation, gonadal development, and exogenous hormone-induced sex reversal remain unknown. In this study, LMB at 15 days post-hatching (dph) were fed diets containing 20 mg/kg of 17a-methyltestosterone (17a-MT) or 30 mg/kg of 17ß-estradiol (17ß-E2) for 60 days, respectively. Serum steroid levels, histological observations of the gonads, and identification of sex-specific markers were employed to screen the gonads of 60-day-old normal female fish (XX-F), normal male fish (XY-M), 17ß-E2 induced pseudo-female fish (XY-F), and 17a-MT-induced pseudo-male fish (XX-M) for transcriptome sequencing in order to uncover genes and pathway involved in the process of sexual reversal. The results from histology and serum sex steroid hormone analysis showed that both 17a-MT and 17ß-E2 were capable of inducing sex reversal of LMB at 15 dph. Transcriptome results revealed a total of 2,753 genes exhibiting differential expression, and the expression pattern of these genes in the gonads of XX-M or XY-F resembled that of normal females or males. The male sex-biased genes that are upregulated in XX-M and downregulated in XY-F are referred to as key genes for male reversal, while the female sex-biased genes that are upregulated in XY-F and downregulated in XX-M are referred to as key genes for female reversal. Finally, 12 differentially expressed genes (DEGs) related to male sex reversal were screened, including star2, cyp17a, cyp11b1, dmrt1, amh, sox9a, katnal1, spata4, spata6l, spata7, spata18 and foxl3. 2DEGs (foxl2a and cyp19a1b) were found to be associated with female sex reversal. The changes in these genes collectively influence the direction of sex differentiation of LMB. Among them, star2, dmrt1 and cyp19a1b with significantly altered expression levels may play potentially crucial role in the process of gender reversal. The expression patterns of 21 randomly selected genes were verified using qRT-PCR which confirmed the reliability and accuracy of the RNA-seq results. These findings not only enhance our understanding of the molecular basis underlying sex reversal but also provide crucial data support for future breeding research on unisexual LMB. [ABSTRACT FROM AUTHOR]

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The sex determination system of largemouth bass ( Micropterus salmoides , LMB) is XX/XY; however, the underlying molecular mechanisms involved in early sex differentiation, gonadal development, and exogenous hormone-induced sex reversal remain unknown. In this study, LMB at 15 days post-hatching (dph) were fed diets containing 20 mg/kg of 17α-methyltestosterone (17α-MT) or 30 mg/kg of 17β-estradiol (17β-E 2 ) for 60 days, respectively. Serum steroid levels, histological observations of the gonads, and identification of sex-specific markers were employed to screen the gonads of 60-day-old normal female fish (XX-F), normal male fish (XY-M), 17β-E 2 induced pseudo-female fish (XY-F), and 17α-MT-induced pseudo-male fish (XX-M) for transcriptome sequencing in order to uncover genes and pathway involved in the process of sexual reversal. The results from histology and serum sex steroid hormone analysis showed that both 17α-MT and 17β-E2 were capable of inducing sex reversal of LMB at 15 dph. Transcriptome results revealed a total of 2,753 genes exhibiting differential expression, and the expression pattern of these genes in the gonads of XX-M or XY-F resembled that of normal females or males. The male sex-biased genes that are upregulated in XX-M and downregulated in XY-F are referred to as key genes for male reversal, while the female sex-biased genes that are upregulated in XY-F and downregulated in XX-M are referred to as key genes for female reversal. Finally, 12 differentially expressed genes (DEGs) related to male sex reversal were screened, including star2 , cyp17a , cyp11b1 , dmrt1 , amh , sox9a , katnal1 , spata4 , spata6l , spata7 , spata18 and foxl3 . 2 DEGs ( foxl2a and cyp19a1b ) were found to be associated with female sex reversal. The changes in these genes collectively influence the direction of sex differentiation of LMB. Among them, star2 , dmrt1 and cyp19a1b with significantly altered expression levels may play potentially crucial role in the process of gender reversal. The expression patterns of 21 randomly selected genes were verified using qRT-PCR which confirmed the reliability and accuracy of the RNA-seq results. These findings not only enhance our understanding of the molecular basis underlying sex reversal but also provide crucial data support for future breeding research on unisexual LMB., Competing Interests: LH was employed by Guangdong Province Liangshi Aquaculture Seed Industry. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhang, Tian, Li, Du, Lei, Zhu, Han and Song.)

The effective detection and tracking of low, small and slow targets is a current hot issue in the radar field. The paper is a sequel to radar weak target detection based on RFT and AMF fusion focus, and concentrates on the tracking of typical UAVs by pulse Doppler radar. The implementations of tracking algorithm mainly include: first, the preprocessing of the plots centroid processing and the multi-frame acceleration estimation has been applied. Then the labeled multi-Bernoulli (LMB) smoother is used for the multi-target tracking. Finally, the final trajectories of targets are output after trajectory continuity judgment and effective trajectory extraction. The LMB smoother can output the smoothed trajectories of targets, therefore, improving the estimation accuracy of distance and velocity. Trajectory continuity judgment and effective track extraction can effectively overcome the problems of breaking trajectory caused by velocity jitter and false trajectory caused by velocity ambiguity. The comprehensive score rate to process multiple test data is 96.67%, which verifies the effectiveness and robustness of the proposed tracking algorithm.

Abstract Unavailability of probiotics in fish digestive system fingerlings is unable to digest and absorb their food properly. The current research was conducted to investigate the influence of probiotics added Linseed meal based (LMB) diet on hematology and carcass composition of Labeo rohita juveniles. Hematological parameters are essential diagnostics used to estimate the health status of fish. The usage of probiotics for fish health improvement is becoming common due to the higher demand for environment-friendly culture system in water. Linseed meal was used as a test ingredient to prepare six experimental test diets by adding probiotics (0, 1, 2, 3, 4 and 5 g/kg) and 1% indigestible chromic oxide for seventy days. According to their live wet weight, five percent feed was given to fingerlings twice a day. Fish blood and carcass samples (Whole body) were taken for hematological and carcass analysis at the end of the experiment. The highest carcass composition (crude protein; 18.72%, crude fat; 8.80% and gross energy; 2.31 kcal/g) was observed in fish fed with test diet II supplemented with probiotics (2 g/kg). Moreover, maximum RBCs number (2.62× 106mm-3), WBCs (7.84×103mm-3), PCV (24.61), platelets (63.85) and hemoglobin (7.87) had also been reported in the fish fingerlings fed on 2 g/kg of probiotics supplemented diet. Results indicated that probiotics supplementation has a critical role in improvement of fingerlings’ body composition and hematological indices. Present findings showed that probiotics supplementation at 2 g/kg level in linseed by-product-based diet was very useful for enhancing the overall performance of L. rohita fingerlings.

Purpose of Review: Burkitt's lymphoma and its leukemic form (Burkitt cell acute lymphoblastic leukemia) are a highly aggressive disease. We review the classification, clinical presentation, histology, cytogenetics, and the treatment of the disease. Recent Findings: Burkitt's lymphoma might be associated with tumor lysis syndrome which is a potentially fatal complication that occurs spontaneously or upon initiation of chemotherapy. Major improvements were made in the treatment of pediatric and adults population using short-course dose-intensive chemotherapy regimens, usually 1 week after a prephase induction. Addition of Rituximab to chemotherapy has become a standard of care. Relapsed/refractory disease has a very poor prognosis and the benefit from autologous/allogeneic hematopoietic stem cell transplant remains uncertain. Summary: Rituximab-based short-course dose-intensive chemotherapy is the standard of care of Burkitt's lymphoma even in the immunodeficiency-related form. [ABSTRACT FROM AUTHOR]

Dissolved oxygen (DO) in the overlying water is important in influencing internal phosphorus (P) release. However, the potentially associative effect of DO on the P adsorption and immobilization by La-modified bentonite (LMB) has not been quantified. This 80-day incubation experiment showed the synergistic effect of DO and LMB in the overlying water, which caused the reduction of dissolved inorganic phosphorus (DIP) by 51% (DO = 5 mg L−1) and 77% (DO = 7 mg L−1) on average, respectively, compared with the DO of 3 mg L−1. In addition, the DIP in the pore water decreased from 1.12 mg P L−1 (control) to 0.014 mg P L−1 (5 mg L−1) and 0.004 mg P L−1 (7 mg L−1). Besides, the Fe2+ and NH4+ concentrations were also reduced significantly in the pore water, suggesting the rise in the redox potential in the sediment, which helped P immobilization. Chemical P-fractionation experiments indicate that the Fe-P reduction in sediment was the most significant, reduced by 14%, followed by NH4Cl-P (12%), causing a reduction by 13% (3 mg L−1), 23% (5 mg L−1) and 27% (7 mg L−1) of mobile P in the surface 7-cm sediment, respectively. However, the released P ions were rapidly adsorbed by the Al ions and Ca ions, as well as their compounds, thereby leading to the obvious rise in inert P in the sediment. Accordingly, it was suggested that DO and LMB had a synergistic effect on external P adsorption and immobilization. [ABSTRACT FROM AUTHOR]

In this paper, we propose a technique for the joint tracking and labeling of multiple extended targets. To achieve multiple extended target tracking using this technique, models for the target measurement rate, kinematic component, and target extension are defined and jointly propagated in time under the generalized labeled multi-Bernoulli filter framework. In particular, we developed a Poisson mixture variational Bayesian model to simultaneously estimate the measurement rate of multiple extended targets and extended target extension was modeled using B-splines. We evaluated our proposed method with various performance metrics. Results demonstrate the effectiveness of our approach. [ABSTRACT FROM AUTHOR]

Reactive astrogliosis is a prominent and ubiquitous reaction of astrocytes to many types of brain injury. Up-regulation of glial fibrillary acidic protein (GFAP) expression and astroglial proliferation are hallmarks of reactive astrogliosis. However, the mechanisms that regulate reactive astrogliosis remain elusive. In the present study, status epilepticus (SE, a prolonged seizure activity) led to reactive astrogliosis showing the increases in GFAP expression and the number of proliferating astrocytes with prolonged extracellular signal receptor-activated kinases 1/2 (ERK1/2) activation and reduced nuclear p27Kip1 level. U0126, an ERK1/2 inhibitor, showed opposite effects. Leptomycin B (LMB), an inhibitor of chromosomal maintenance 1 (CRM1), attenuated nucleocytoplasmic p27Kip1 export and astroglial proliferation, although it up-regulated ERK1/2 phosphorylation and GFAP expression. Roscovitine ameliorated the reduced nuclear p27Kip1 level and astroglial proliferation without changing GFAP expression and ERK1/2 phosphorylation. U0126 aggravated SE-induced astroglial apoptosis in the molecular layer of the dentate gyrus that was unaffected by LMB and roscovitine. In addition, U0126 exacerbated SE-induced neuronal death, while LMB mitigated it. Roscovitine did not affect SE-induced neuronal death. The present data elucidate for the first time the roles of nucleocytoplasmic p27Kip1 transport in ERK1/2-mediated reactive astrogliosis independent of SE-induced neuronal death and astroglial apoptosis. Therefore, our findings suggest that nucleocytoplasmic p27Kip1 export may be required for ERK1/2-mediated astroglial proliferation during reactive astrogliosis, and that nuclear p27Kip1 entrapment may be a potential therapeutic strategy for anti-proliferation in reactive astrocytes.

Reactive astrogliosis is a prominent and ubiquitous reaction of astrocytes to many types of brain injury. Up-regulation of glial fibrillary acidic protein (GFAP) expression and astroglial proliferation are hallmarks of reactive astrogliosis. However, the mechanisms that regulate reactive astrogliosis remain elusive. In the present study, status epilepticus (SE, a prolonged seizure activity) led to reactive astrogliosis showing the increases in GFAP expression and the number of proliferating astrocytes with prolonged extracellular signal receptor-activated kinases 1/2 (ERK1/2) activation and reduced nuclear p27Kip1 level. U0126, an ERK1/2 inhibitor, showed opposite effects. Leptomycin B (LMB), an inhibitor of chromosomal maintenance 1 (CRM1), attenuated nucleocytoplasmic p27Kip1 export and astroglial proliferation, although it up-regulated ERK1/2 phosphorylation and GFAP expression. Roscovitine ameliorated the reduced nuclear p27Kip1 level and astroglial proliferation without changing GFAP expression and ERK1/2 phosphorylation. U0126 aggravated SE-induced astroglial apoptosis in the molecular layer of the dentate gyrus that was unaffected by LMB and roscovitine. In addition, U0126 exacerbated SE-induced neuronal death, while LMB mitigated it. Roscovitine did not affect SE-induced neuronal death. The present data elucidate for the first time the roles of nucleocytoplasmic p27Kip1 transport in ERK1/2-mediated reactive astrogliosis independent of SE-induced neuronal death and astroglial apoptosis. Therefore, our findings suggest that nucleocytoplasmic p27Kip1 export may be required for ERK1/2- mediated astroglial proliferation during reactive astrogliosis, and that nuclear p27Kip1 entrapment may be a potential therapeutic strategy for anti-proliferation in reactive astrocytes. [ABSTRACT FROM AUTHOR]

CRM1 (chromosome maintenance region 1, Exportin 1) binds to nuclear export signals and is required for nucleocytoplasmic transport of a large variety of proteins and RNP complexes. Leptomycin B (LMB), the first specific inhibitor of CRM1 identified, binds covalently to cysteine 528 in the nuclear export signal binding region of CRM1 leading to the inhibition of protein nuclear export. Although the biochemical mechanisms of action of CRM1 inhibitors such as LMB are well studied, the subcellular effects of inhibition on CRM1 are unknown. We have found that LMB causes CRM1 to redistribute from the nucleus to the cytoplasm in A549 cells. A significant decrease in nuclear CRM1 coupled with an increase in cytoplasmic CRM1 was sustained for up to 4 h, while there was no change in total CRM1 protein in fractionated cells. Cells expressing an LMB insensitive HA-tagged CRM1-C528S protein were unaffected by LMB treatment, whereas HA-tagged wildtype CRM1 redistributed from the nucleus to the cytoplasm with LMB treatment, similar to endogenous CRM1. GFP-tagged CRM1 protein microinjected into the cytoplasm of A549 cells distributed throughout the cell in untreated cells remained primarily cytoplasmic in LMB-treated cells. Upon nuclear microinjection, GFP-CRM1 translocated to and accumulated in the cytoplasm of LMB-treated cells. Thus, LMB binds to CRM1 and causes its redistribution to the cytoplasm by inhibiting its nuclear import. Decreasing the nuclear availability of CRM1 likely contributes to the accumulation of CRM1 cargo proteins in the nucleus, suggesting a new mechanism of action for LMB. [ABSTRACT FROM AUTHOR]

Fluorescent protein biosensors are powerful cellular systems biology tools for dissecting the complexity of cellular processes with high spatial and temporal resolution. As regulated nucleo-cytoplasmic transport is crucial for the modulation of numerous (patho)physiological cellular responses, a detailed understanding of its molecular mechanism would open up novel options for a rational manipulation of the cell. In contrast to genetic approaches, we here established and employed high-content cellular translocation biosensors applicable for dissecting nuclear export by chemicogenomics. A431 cell lines, stably expressing a translocation biosensor composed of glutathione S-transferase, GFP and a rational combination of nuclear import and export signals, were engineered by antibiotic selection and flow cytometry sorting. Using an optimized nuclear translocation algorithm, the translocation response could be robustly quantified on the Cellomics Arrayscan® VTI platform. Subsequent to assay optimization, the assay was developed into a higher density 384-well format high-content assay and employed for the screening of the 17K ChemBioNet compound collection. This library was selected on the basis of a genetic algorithm used to identify maximum common chemical substructures in a database of annotated bioactive molecules and hence, is well-placed in the chemical space covered by bioactive compounds. Automated multiparameter data analysis combined with visual inspection allowed us to identify and to rationally discriminate true export inhibitors from false positives, which included fluorescent compounds or cytotoxic substances that dramatically affected the cellular morphology. A total of 120 potential hit compounds were selected for Cellomics Arrayscan® VTI based rescreening. The export inhibitory activity of 20 compounds effective at concentrations < 25 μM were confirmed by fluorescence microscopy in several cell lines. Interestingly, kinetic analysis allowed the identification of inhibitors capable to interfere with the export receptor CRM1-mediated nuclear export not only in an irreversible, but also in a reversible fashion. In sum, exploitation of biosensor based screening allows the identification of chemicogenomic tools applicable for dissecting nucleo-cytoplasmic transport in living cells.

Tuning the optical response of lithium magnesium borate glass via controlled doping of rare earth ions is the key issue in photonic devices. Glasses with composition 30Li 2 O–(60− x )B 2 O 3 –10MgO- x Er 2 O 3 , where 0 ≤ x ≤ 1 are prepared by conventional melt-quenching technique. The X-ray diffraction (XRD) pattern confirms the amorphous nature of all samples. Fourier transform infrared (FTIR) spectra reveal the presence of BO 3 and BO 4 local structure unit. The physical parameters, such as the direct and indirect optical energy band gap, oscillator strength, refractive index, ion concentration, Polaron radius, molar volume and inter-nuclear distance are calculated and analyzed. The room temperature UV–vis–IR spectra comprised of ten absorption bands centered at 1523, 973, 796, 650, 550, 522, 486, 447, 406, 373 nm corresponding to the transitions from the ground state to 4 I 13/2 , 4 I 11/2 , 4 I 9/2 , 4 F 9/2 , 4 S 3/2 , 2 H 11/2 , 4 F 7/2 , ( 4 F 5/2 + 4 F 3/2 ), 2 G(1) 9/2 , 4 G 11/2 excited states, respectively. The peak evidenced at 522 nm is due to hypersensitive transition. The up-conversion spectra exhibits three emission peaks centered at 509, 547 and 656 nm. All the emission bands (green and red) at 0.5 mol% of Er 3+ shows a significant enhancement in the intensity attributed to the energy transfer from Mg 2+ to the Er 3+ ion. Our results suggest that these glasses can be nominated for solid state lasers. [ABSTRACT FROM AUTHOR]

Enhancing the up-conversion efficiency of borate glass via optimized doping of rare earth ions is an ever-ending quest in lasing glass. Neodymium (Nd 3+ ) doped Lithium Magnesium Borate (LMB) glasses are prepared using the melt-quenching method. X-ray diffraction (XRD), Fourier transformed infrared (FTIR), UV–Vis–NIR absorption and Photoluminescence (PL) spectroscopic characterizations are made to examine the influence of Nd 3+ concentration on physical properties and optical properties. Nd 3+ contents dependent density, molar volume, refractive index, ion concentration, Polaron radius, inter nuclear distance, field strength, energy band gap and oscillator strength are calculated. XRD patterns confirm the amorphous nature of all glasses and the FTIR spectra reveal the presence of BO 3 and BO 4 functional groups. UV–Vis–IR spectra exhibit ten prominent bands centered at 871, 799, 741, 677, 625, 580, 522, 468, 426, 349 nm corresponding to the transitions from the ground state to 4 F 3/2 , ( 4 F 5/2 + 2 H 9/2 ), ( 4 F 7/2 + 4 S 3/2 ), 4 F 9/2 , 2 H 11/2 , ( 4 G 5/2 + 2 G 7/2 ), ( 2 K 13/2 + 4 G 7/2 + 4 G 9/2 ), ( 2 G 9/2 + 2 D 3/2 + 2 P 3/2 ), ( 2 P 1/2 + 2 D 5/2 ), ( 4 D 3/2 + 4 D 5/2 ) excited states, respectively. A hyper-sensitive transition related to ( 4 G 5/2 + 2 G 7/2 ) level is evidenced at 580 nm. The room temperature up-conversion emission spectra at 800 nm excitation displays three peaks centered at 660, 610 and 540 nm. Glass with 0.5 mol% of Nd 3+ showing an emission enhancement by a factor to two is attributed to the energy transfer between Mg 2+ and Nd 3+ ions. Our results suggest that these glasses can be nominated for solid state lasers and other photonic devices. [ABSTRACT FROM AUTHOR]

As the staff of the MRC Laboratory of Molecular Biology settle into their new building in Cambridge, its director Hugh Pelham explains the challenges of living up to its prestigious past.

Abstract: Streptococcus suis (S. suis) is an important zoonotic pathogen that causes multiple diseases in both pigs and humans. Many studies suggest that Streptococcus utilizes host extracellular matrix proteins, including laminin, for adhesion and invasion of host cells. Recently, we identified a putative Lmb protein (CDS 0330) of a highly virulent strain of S. suis (serotype 2). In this study, we characterized the ability of CDS 0330 to bind human laminin, and evaluated the protective efficacy of a recombinant protein vaccine. Bioinformatic analysis revealed that both the amino acid sequence and tertiary structure of CDS 0330 were similar to Lmb proteins in other Streptococcus. In addition, the sequence of CDS 0330 was present in the genomes of 26 of the 38 sequenced streptococci species, indicating an early origin and conservation of this gene. Particularly, all 17 sequenced S. suis genomes, regardless of serotype or geographic origin, contained CDS 0330 gene in their genome with a minimum pair-wise amino acid identity of 92%. PCR amplification revealed that CDS 0330 gene is distributed throughout 35 S. suis serotypes in the lmb-htp format. Flow cytometry analysis confirmed that CDS 0330 was expressed on the cell surface of S. suis, and ELISA revealed the recombinant CDS 0330 protein could bind laminin in vitro. Finally, vaccinating mice with recombinant CDS 0330 protein significantly prolonged survival after S. suis infection. Together, these data reveal that CDS 0330 is a laminin binding protein of S. suis 2, and open new avenues for preventing S. suis 2 infection. [Copyright &y& Elsevier]

Trafficking of proteins between the cytoplasm and nucleus occurs exclusively across the nuclear pore complex of eucaryotic cells. Fundamental aspects of this process affect temporal and spatial parameters, the latter carried out by specific import [nuclear localization sequence (NLS)] and export [nuclear export sequence (NES)] sequences. In this study, we focused on the adaptation of a protein heterodimerization assay to kinetically measure Crm1-mediated nuclear export in living cells using the rapalog AP21967, a heterodimerizing agent and NLS- and NES-containing fusion proteins equipped with distinct AP21967-specific binding motifs. In HeLa cells, we observed rapid nuclear export of the NLS-containing fusion protein in the presence of AP21967, with the extent of this process being a function of the number of AP21967-binding motifs. AP21967-induced nuclear export was specifically inhibited by the Crm1-binding molecule leptomycin B. Half maximal export was achieved after ∼ 10 min. We further applied protein heterodimerization in HeLa cells to study induced NLS-mediated nuclear import. Only in the presence of heterodimerizer AP21967 nuclear import of a cytoplasmically localizing fusion protein was observed. Induced protein heterodimerization is thus a valuable tool to quantitatively study nucleocytoplasmic protein trafficking in cultured cells, in a non-invasive, time-saving manner. [ABSTRACT FROM AUTHOR]

Fluorescent protein biosensors are powerful cellular systems biology tools for dissecting the complexity of cellular processes with high spatial and temporal resolution. As regulated nucleo-cytoplasmic transport is crucial for the modulation of numerous (patho)physiological cellular responses, a detailed understanding of its molecular mechanism would open up novel options for a rational manipulation of the cell. In contrast to genetic approaches, we here established and employed high-content cellular translocation biosensors applicable for dissecting nuclear export by chemicogenomics. A431 cell lines, stably expressing a translocation biosensor composed of glutathione Stransferase, GFP and a rational combination of nuclear import and export signals, were engineered by antibiotic selection and flow cytometry sorting. Using an optimized nuclear translocation algorithm, the translocation response could be robustly quantified on the Cellomics Arrayscan® VTI platform. Subsequent to assay optimization, the assay was developed into a higher density 384-well format high-content assay and employed for the screening of the 17K ChemBioNet compound collection. This library was selected on the basis of a genetic algorithm used to identify maximum common chemical substructures in a database of annotated bioactive molecules and hence, is well-placed in the chemical space covered by bioactive compounds. Automated multiparameter data analysis combined with visual inspection allowed us to identify and to rationally discriminate true export inhibitors from false positives, which included fluorescent compounds or cytotoxic substances that dramatically affected the cellular morphology. A total of 120 potential hit compounds were selected for Cellomics Arrayscan® VTI based rescreening. The export inhibitory activity of 20 compounds effective at concentrations < 25 μM were confirmed by fluorescence microscopy in several cell lines. Interestingly, kinetic analysis allowed the identification of inhibitors capable to interfere with the export receptor CRM1-mediated nuclear export not only in an irreversible, but also in a reversible fashion. In sum, exploitation of biosensor based screening allows the identification of chemicogenomic tools applicable for dissecting nucleo-cytoplasmic transport in living cells. [ABSTRACT FROM AUTHOR]

Survivin appears to function as an apoptosis inhibitor and a regulator of cell division during development and tumorigenesis. Here we report the molecular characterization of the nucleocytoplasmic transport of survivin mid its potential implications for tumorigenesis. We identified an evolutionary conserved Crm1-dependent nuclear export signal (NES) in survivin. In dividing cells, the NES is essential for tethering survivin and the survivin/Aurora-B kinase complex to the mitotic machinery, which in turn appears to be essential for proper cell division. In addition, export seems to be required for the cytoprotective activity of survivin, as export-deficient survivin fails to protect tumor cells against chemo- and radiotherapy-induced apoptosis. These findings appear to be clinically relevant since preferential nuclear localization of survivin correlated with enhanced survival in colorectal cancer patients. Targeting survivin's nuclear export by the application of NES-specific antibodies promoted its nuclear accumulation and inhibited its cytoprotective function. We demonstrate that nuclear export is essential for the biological activity of survivin and promote the identification of molecular decoys to specifically interfere with survivin's nuclear export as potential anticancer therapeutics. [ABSTRACT FROM AUTHOR]

Survivin appears to function as a regulator of cell division and as an apoptosis inhibitor in many species. Here, we characterized the nucleocytoplasmic transport of mouse survivin140, and its splice variants survivin121 and survivin40. We show that the dynamic intracellular localization of survivin140 is mediated by a Crm1-dependent nuclear export signal (NES) present also in survivin121, but absent in survivin40. In contrast, neither survivin nor survivin splice variants contain an active nuclear import signal and seem to enter the nucleus by passive diffusion. The activity of the NES is required for survivin-mediated protection against cell death inducing stimuli and influences protein degradation. During mitosis, NES-deficient survivin variants fail to correctly localize to the mitotic machinery and promote proper cell division. In vivo and in vitro protein interaction assays show that survivin140 and survivin121 as well as their export-deficient mutants are able to form homo- as well as heterodimers. The trans-dominant negative phenotype observed upon expression of export-deficient survivin appears, therefore, to be mediated by the formation of inactive survivin heterodimers. The survivin–Crm1 axis is essential for the biological activities of murine survivin, and mouse models will allow investigating its functional implications during development and tumorigenesis. [ABSTRACT FROM AUTHOR]

Dynamic subcellular localization is an important regulatory mechanism for many proteins. cIAP1 and cIAP2 are two closely related members of inhibitor of apoptosis (IAP) family that play a role both as caspase inhibitors and as mediators of tumor necrosis factor (TNF) receptor signaling. Here, we report that cIAP1 and cIAP2 are nuclear shuttling proteins, whose subcellular localization is mediated by the CRM1-dependent nuclear export pathway. Blocking export with leptomycin B induces accumulation of both endogenous cIAP1 and epitope-tagged cIAP1 and cIAP2 in the nucleus of human cancer cells. We have identified a new CRM1-dependent leucine-rich nuclear export signal (NES) in the linker region between cIAP1 BIR2 and BIR3 repeats. Mutational inactivation of the NES, which is not conserved in cIAP2, reduces cIAP1 nuclear export. Forced relocation of cIAP1 to the nucleus did not significantly alter its ability to prevent apoptosis. Interestingly, co-expression experiments showed that the cIAP1 and cIAP2-interacting protein TNF receptor-associated factor 2 (TRAF2) plays an important role as regulator of IAP nucleocytoplasmic localization, by preventing nuclear translocation of cIAP1 and cIAP2. TRAF2-mediated cytoplasmic retention of cIAP1 was reduced upon TNFα treatment. Our results identify molecular mechanisms that contribute to regulate the subcellular localization of cIAP1 and cIAP2. Translocation between different cell compartments may add a further level of control for cIAP1 and cIAP2 activity. [Copyright &y& Elsevier]

The Karyopherin protein CRM1 or XPO1 is the major nuclear export receptor that regulates nuclear exit of thousands of macromolecules in the cell. CRM1 recognizes protein cargoes by binding to their 8-15 residue-long nuclear export signals (NESs). A ternary CRM1-Ran-RanBP1 complex engineered to be suitable for crystallization has enabled structure determination by X-ray crystallography of CRM1 bound to many NES peptides and small-molecule inhibitors. Here, we present a protocol for the purification of the individual proteins, formation of the ternary CRM1-Ran-RanBP1 complex and crystallization of this complex for X-ray crystallography., (© 2022. Springer Science+Business Media, LLC, part of Springer Nature.)

Herein, we discuss the impact of water pH on mobile and organic phosphorous (P), as well as the kinetics and efficacy of P responses following LMB amendment at the sediment-water interface. For this study, we conducted a microcosm incubation experiment over a 90-day period during which the pH of the overlying water was continuously adjusted. Over the course of this experimental period, one- and two-dimensional soluble and labile P sediment profiles were assessed using high-resolution Peeper (HR-Peeper) and diffusive gradients in thin-film (DGT) probes. These analyses revealed that rising water column pH can induce significant increases in SRP and labile P concentrations, as well as substantial changes in levels of inorganic P (IP) and organic P (OP) species. The P release kinetics under different pH conditions were calculated using a DGT-induced flux in sediments (DIFS) model. We found that sediment still serves as a P pool following LMB addition, ultimately resupplying soluble P to the overlying water. However, P release became increasingly challenging with reduction in pH. Together, our results provided direct evidence regarding the impact of pH on efficiency of the LMB capping of phosphorous, potentially guiding future studies of the engineering application of LMB. [ABSTRACT FROM AUTHOR]

Lithium metal is a promising anode candidate for high-energy-density secondary batteries due to its high theoretical capacity and low electrochemical potential, but the uncontrolled lithium dendrite growth causing the poor cycling performance and safety concerns becomes the main drawback in the lithium metal batteries (LMBs). This study presents a phase-field method for modeling the lithium dendrite formation in LMBs. In this model, the electrochemical kinetics is described by a modified Butler–Volmer equation for deposition non-uniformity, and the Nernst–Planck equations and charge neutrality condition are used to solve the electric potential field and the ionic concentration fields. Our simulation results show that this model is able to accurately describe the growth of mossy dendrites, which is missing in previous phase-field works. We found that the lithium dendrite growth is strongly affected by the deposition non-uniformity originated from the competition of SEI growth and lithium growth and the charge conditions. Under high deposition rates, the depletion of the lithium ionic concentration takes place near the anode, and the dendrite growth undergoes a transition from the reaction-limited mossy growth to the diffusion-limited fractal growth. • A phase-field model for SEI-influenced lithium dendrites was developed. • A modified Butler–Volmer kinetics for SEI/dendrite growth competition is proposed. • The model bridges the length-scale gap between SEI and lithium dendrites. • Large scale simulations for moderate to high deposition rates were performed. • The model captures mossy/fractal dendrite behavior observed in experiments. [ABSTRACT FROM AUTHOR]

In this work, we study the effect on the electric potential difference (EPD) of a liquid metal (LM) flow confined in a cuboid vessel. The interaction between a magnetic field (MF) and an electric current density (j 0) along the ascending axial direction generates a Lorentz force that stirs the LM producing an MHD flow. The MF is produced by either a single or a pair of NdFeB dipolar permanent magnets. j 0 is in the range of 384 to 1153 A m-2. Experimental characterization is done by measuring velocity components using the ultrasound Doppler velocimetry (UDV) technique and measurements of the EPD in terminals of the vessel. Additionally, an MHD model is solved using the COMSOL Multiphysics software. A good agreement between measurements and simulations for the magnetic and velocity fields is obtained. Furthermore, an order of magnitude analysis for the characteristic velocity of the flow and EPD is presented. Comparing with a configuration without magnets, the EPD when one magnet is used reaches 16.8 % higher values. In contrast, for a pair of magnets values are 22.9 % smaller. This increasing/decreasing behavior for the EPD with the flow patterns could be of interest to applications such as liquid metal batteries. • A Lorentz force stirs the GaInSn alloy confined in a cuboid vessel. • NdFeB magnets and currents in the range of 384 to 1153 A m-2 are considered. • Velocity profiles are measured with the UDV technique. • Numerical simulations with COMSOL Multiphysics are performed. • The electric potential difference depends on the flow behavior. [ABSTRACT FROM AUTHOR]

LMB is a widely utilized material for the management of sediment-derived phosphorus (P) in eutrophic lakes. However, the properties of organic P at the sediment-water interface and the effect of LMB on organic P fractions in sediments are still unclear. The batch studies reported here indicate that LMB has good adsorption properties toward organic P (Glucose-6-phosphate), and the tentative adsorption mechanism relies on chemisorption. Laboratory microcosmic experiments were conducted to study the immobilization effect of LMB on the organic P in water and sediments. The results indicated that the concentration of total P, labile P and organic P in overlying water and pore water could be effectively reduced by LMB capping of sediments. After treatment, the optimal immobilization effects appeared on the 7th day and until the 60th day. However, the fractions of organic P change during the capping time. Active organic P eluted with NaHCO 3 transforms into moderately labile or non-labile P through the physical and chemical processes, as well as microbial action. Microbial community analysis showed that the addition of LMB had inhibitory effect on the phosphorus-solubilizing bacteria, which also affected the transformation between various forms of organic P. This study provided new insights of LMB in situ capping of organic P and the mechanisms of the migration and cycling of internal organic P, which is beneficial for the management of eutrophic lakes. Unlabelled Image • Characteristics of LMB on organic P was first observed. • Zr-oxide DGT was used for 2D measurement of labile P in the vertical sediment. • LMB has effect on the species of organic P during different incubation time. • Microbial communities are observed and be found changed after amendment of LMB. [ABSTRACT FROM AUTHOR]

The purpose of this study is to promote a new way of application composite materials to restore eutrophic waters. A new sustainable way of application is based on the "teabag" method, in which materials were placed in water-permeable bags and immersed in the water column in order to sorb phosphate—one of the main contributory element for the eutrophication problem. Particularly, the two composites materials of Phoslock™ (lanthanum-modified bentonite, LMB) and Bephos™ (Fe-modified bentonite, f-MB) were tested and bench-scale batch experiments were employed to investigate their sorption efficiency in the forms of slurry and teabag. The adsorption kinetics and the relevant adsorption isotherms were deployed, while the effect of the materials on turbidity and their aging were also investigated. Experimental results showed that Phoslock™ and Bephos™ (as teabag), being applied at initial concentration range: 0.05–5 mg/L, they sustained a maximum adsorption capacity of 7.80 mg/g and 25.1 mg/g, respectively, which are considered sufficient rates for P concentrations reported at natural aquatic ecosystems. At the same time this new method did not cause turbidity in the water column, since the material was not released into the water, thus, preventing potential harmful consequences for the living organisms. Moreover, the "teabag" method prevents the material to cover the lake bottom, avoiding the phenomenon of smothering of benthos. Βy teabag method, the materials can be collected for further applicability as soil improver or crops fertilizer. Finally, it was argued that the possibility to recycle LMB and f-MB materials for agricultural use is of paramount importance, sustaining also positive impacts on sustainable ecology and on the routes of circular economy (CE). [ABSTRACT FROM AUTHOR]