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Human patients carrying genetic mutations in RNA binding motif 20 (RBM20) develop a clinically aggressive dilated cardiomyopathy (DCM). Genetic mutation knockin (KI) animal models imply that altered function of the arginine-serine-rich (RS) domain is crucial for severe DCM. To test this hypothesis, we generated an RS domain deletion mouse model (Rbm20ΔRS). We showed that Rbm20ΔRS mice manifested DCM with mis-splicing of RBM20 target transcripts. We found that RBM20 was mis-localized to the sarcoplasm in Rbm20ΔRS mouse hearts and formed RBM20 granules similar to those detected in mutation KI animals. In contrast, mice lacking the RNA recognition motif showed similar mis-splicing of major RBM20 target genes but did not develop DCM or exhibit RBM20 granule formation. Using in vitro studies with immunocytochemical staining, we demonstrated that only DCM-associated mutations in the RS domain facilitated RBM20 nucleocytoplasmic transport and promoted granule assembly. Further, we defined the core nuclear localization signal (NLS) within the RS domain of RBM20. Mutation analysis of phosphorylation sites in the RS domain suggested that this modification may be dispensable for RBM20 nucleocytoplasmic transport. Collectively, our findings revealed that disruption of RS domain–mediated nuclear localization is crucial for severe DCM caused by NLS mutations.

To address the issue of current garbage cleanup vessels being limited to performing garbage cleaning operations in the ocean, without the capability of transferring the garbage from the ocean to the land, this paper presents a spiral-propulsion amphibious intelligent robot for land garbage cleaning and sea garbage cleaning. The design solution is as follows. A mechanical structure based on a spiral drum is proposed. The interior of the spiral drum is hollow, providing buoyancy, allowing the robot to travel both on marshy, tidal flats and on the water surface, in conjunction with underwater thrusters. Additionally, a mechanical-arm shovel is designed, which achieves two-degrees-of-freedom movement through a spiral spline guide and servo, facilitating garbage collection. Our experimental results demonstrated that the robot exhibits excellent maneuverability in marine environments and on beach, marsh, and tidal flat areas, and that it collects garbage effectively.

In remote sensing images, small objects have too few discriminative features, are easily confused with background information, and are difficult to locate, leading to a degradation in detection accuracy when using general object detection networks for aerial images. To solve the above problems, we propose a remote sensing small object detection network based on the attention mechanism and multi-scale feature fusion, and name it AMMFN. Firstly, a detection head enhancement module (DHEM) was designed to strengthen the characterization of small object features through a combination of multi-scale feature fusion and attention mechanisms. Secondly, an attention mechanism based channel cascade (AMCC) module was designed to reduce the redundant information in the feature layer and protect small objects from information loss during feature fusion. Then, the Normalized Wasserstein Distance (NWD) was introduced and combined with Generalized Intersection over Union (GIoU) as the location regression loss function to improve the optimization weight of the model for small objects and the accuracy of the regression boxes. Finally, an object detection layer was added to improve the object feature extraction ability at different scales. Experimental results from the Unmanned Aerial Vehicles (UAV) dataset VisDrone2021 and the homemade dataset show that the AMMFN improves the APs values by 2.4% and 3.2%, respectively, compared with YOLOv5s, which represents an effective improvement in the detection accuracy of small objects.

Abstract RNA binding motif 20 (RBM20) cardiomyopathy has been detected in approximately 3% of populations afflicted with dilated cardiomyopathy (DCM). It is well conceived that RBM20 cardiomyopathy is provoked by titin isoform switching in combination with resting Ca2+ leaking. In this study, we characterized the cardiac function in Rbm20 knockout (KO) rats at 3‐, 6‐, 9‐, and 12‐months of age and examined the effect of the ryanodine receptor stabilizer S107 on resting intracellular levels and cardiomyocyte contractile properties. Our results revealed that even though Rbm20 depletion promoted expression of larger titin isoform and reduced myocardial stiffness in young rats (3 months of age), the established DCM phenotype required more time to embellish. S107 restored elevated intracellular Ca2+ to normal levels and ameliorated cardiomyocyte contractile properties in isolated cardiomyocytes from 6‐month‐old Rbm20 KO rats. However, S107 failed to preserve cardiac homeostasis in Rbm20 KO rats at 12 months of age, unexpectedly, likely due to the existence of multiple pathogenic mechanisms. Taken together, our data suggest the therapeutic promises of S107 in the management of RBM20 cardiomyopathy.

Leydig cells (LCs) are the predominant cells of androgen production, which plays key roles in spermatogenesis and maintaining male secondary sexual characteristics. Abnormal development of LCs affects androgen levels in vivo, affects fertility and may even lead to infertility. Little is known about the regulation mechanism on LCs’ development and maturation in domestic animals, especially the regulation of non-coding RNAs. In this study, we continued to dig deeper in the previous RNA-seq data of porcine LCs from our group, combined with detecting the expression profiles in different tissues and different types of cells in the testis, to screen out candidate microRNAs (miRNAs) that may affect the regulation of LCs. A total of two miRNAs, ssc-miR-21-5p and ssc-miR-615 (“ssc” is omitted below), were finally determined. After overexpression and interference of miRNAs in vitro, the effects of candidate miRNAs on the proliferation and apoptosis of TM3 (mouse Leydig cell line) were explored. The results showed that miR-21-5p led to a decrease in TM3 cell density and p53 (apoptosis related protein) expression. Meanwhile, miR-21-5p decreased EdU positive cell numbers, but increased TUNEL positive cell numbers, suggesting miR-21-5p could inhibit proliferation and promote apoptosis. Conversely, miR-615 could increase TM3 cell density. Western blot and TUNEL assay indicated miR-615 inhibited apoptosis, but had no effect on proliferation. In addition, Sox5 was identified a potential target gene of these two miRNAs by Dual-Luciferase reporter system assay. Our findings about functions of miRNAs in TM3 and the mapping of miRNAs-target gene regulatory network would provide an important basis for the further elucidation of miRNAs in regulating pig LCs.

The adult Leydig cells (ALCs), originated from stem Leydig cells (SLCs), can secrete testosterone which is essential for germ cell development and sexual behavior maintenance. As a synthetic compound, ethane dimethane sulfonate (EDS), a well-known alkylating agent, has been reported to specifically ablate ALCs. In this study, EDS was verified to ablate differentiated pig LCs by experiments. Subsequently, the primary isolated pig LCs (containing SLCs and differentiated LCs) and EDS-treated LCs (almost exclusively SLCs) were collected for RNA-seq 4,904 genes and 15 miRNAs were differently expressed between the two groups. Down-regulated genes in the EDS-treated group were mainly related to steroid hormone biosynthesis. The highest up-regulation miRNAs was miR-205 after EDS treatment. Additionally, miR-205 was expressed more highly in pig SLCs clones compared with differentiated LCs. Through qRT-PCR, western blot (WB), TUNEL, EDU and flow cytometry, miR-205 was found to induce cell apoptosis, but did not affect proliferation or differentiation in both TM3 and GC-1spg mouse cell lines. Through luciferase reporter assays and WB, RAP2B was identified as a target gene of miR-205. Besides, overexpression of miR-205 inhibited the expressions of PI3K, Akt and p-AKT. All these findings were helpful for elucidating the regulation mechanism in pig LCs.

POU (Pit-Oct-Unc) class 1 homeobox 1 (POU1F1, or Pit-1) is a transcription factor that directly regulates pituitary hormone-related genes, as well as affects the reproduction and growth in mammals. Thus, POU1F1 gene was investigated as a candidate gene for litter size and growth performance in goats. In the current study, using direct DNA sequencing, c.682G > T, c.723T > G and c.837T > C loci were genotyped in Shaanbei white cashmere (SBWC) goats (n = 609), but c.876 + 110T > C was monomorphic. Besides, the c.682G > T locus was first identified by HinfI (Haemophilus influenzae Rf) restriction endonuclease. Association analysis results showed that the c.682G > T, c.837T > C loci and diplotypes were significantly associated with goat litter size (p < 0.05). The positive genotypes were GT and TT for the two SNPs, respectively, and the optimal diplotype was H3H7 (GTTT-TTTT). On the other hand, the c.682G > T, c.723T > G and c.837T > C strongly affected growth traits and body measurement indexes in SBWC goats (p < 0.05). The positive genotypes or allele of these SNPs were GT, G and TT, respectively. Additionally, the goats with H3H7 diplotype also had a greater growth status than others (p < 0.05). Here, individuals with same genotype had both a better litter size and growth traits, showing a positive correlation between these economic traits. Meanwhile, the positive genotypes of four SNPs were combined to obtain the optimal diplotype, which was also H3H7. These SNPs, especially the diplotype, could be used for the genomic selection of excellent individuals with a greater litter size and better growth status in goat breeding.

The human body is often occluded by a variety of obstacles in the monitoring system, so occluded person re-identification is still a long-standing challenge. Recent methods based on pose guidance or external semantic clues have improved the representation and related performance of features; there are still problems, such as weak model representation and unreliable semantic clues. To solve the above problems, we proposed a feature extraction network, named shared feature fusion with pose-guided and unsupervised semantic segmentation (SFPUS). This network will extract more discriminative features and reduce the occlusion noise on pedestrian matching. Firstly, the multibranch joint feature extraction module (MFE) is used to extract feature sets containing pose information and high-order semantic information. This module not only provides robust extraction capabilities but can also precisely segment occlusion and the body. Secondly, in order to obtain multiscale discriminant features, the multiscale correlation feature matching fusion module (MCF) is used to match the two feature sets, and the Pose–Semantic Fusion Loss is designed to calculate the similarity of the feature sets between different modes and fuse them into a feature set. Thirdly, to solve the problem of image occlusion, we use unsupervised cascade clustering to better prevent occlusion interference. Finally, performances of the proposed method and various existing methods are compared on the Occluded-Duke, Occluded-ReID, Market-1501 and Duke-MTMC datasets. The accuracy of Rank-1 reached 65.7%, 80.8%, 94.8% and 89.6%, respectively, and the mAP accuracy reached 58.8%, 72.5%, 91.8% and 80.1%. The experiment results demonstrate that our proposed SFPUS holds promising prospects and performs admirably compared with state-of-the-art methods. [ABSTRACT FROM AUTHOR]

To address the issue of current garbage cleanup vessels being limited to performing garbage cleaning operations in the ocean, without the capability of transferring the garbage from the ocean to the land, this paper presents a spiral-propulsion amphibious intelligent robot for land garbage cleaning and sea garbage cleaning. The design solution is as follows. A mechanical structure based on a spiral drum is proposed. The interior of the spiral drum is hollow, providing buoyancy, allowing the robot to travel both on marshy, tidal flats and on the water surface, in conjunction with underwater thrusters. Additionally, a mechanical-arm shovel is designed, which achieves two-degrees-of-freedom movement through a spiral spline guide and servo, facilitating garbage collection. Our experimental results demonstrated that the robot exhibits excellent maneuverability in marine environments and on beach, marsh, and tidal flat areas, and that it collects garbage effectively. [ABSTRACT FROM AUTHOR]

In remote sensing images, small objects have too few discriminative features, are easily confused with background information, and are difficult to locate, leading to a degradation in detection accuracy when using general object detection networks for aerial images. To solve the above problems, we propose a remote sensing small object detection network based on the attention mechanism and multi-scale feature fusion, and name it AMMFN. Firstly, a detection head enhancement module (DHEM) was designed to strengthen the characterization of small object features through a combination of multi-scale feature fusion and attention mechanisms. Secondly, an attention mechanism based channel cascade (AMCC) module was designed to reduce the redundant information in the feature layer and protect small objects from information loss during feature fusion. Then, the Normalized Wasserstein Distance (NWD) was introduced and combined with Generalized Intersection over Union (GIoU) as the location regression loss function to improve the optimization weight of the model for small objects and the accuracy of the regression boxes. Finally, an object detection layer was added to improve the object feature extraction ability at different scales. Experimental results from the Unmanned Aerial Vehicles (UAV) dataset VisDrone2021 and the homemade dataset show that the AMMFN improves the A P s values by 2.4% and 3.2%, respectively, compared with YOLOv5s, which represents an effective improvement in the detection accuracy of small objects. [ABSTRACT FROM AUTHOR]

(1) Background: RNA binding motif 20 (RBM20) regulates mRNA splicing specifically in muscle tissues. Missense mutations in the arginine/serine (RS) domain of RBM20 lead to abnormal gene splicing and have been linked to severe dilated cardiomyopathy (DCM) in human patients and animal models. Interestingly, many of the reported DCM-linked missense mutations in RBM20 are in a highly conserved RSRSP stretch within the RS domain. Recently, it was found that the two Ser residues within this stretch are constitutively phosphorylated, yet the identity of the kinase(s) responsible for phosphorylating these residues, as well as the function of RSRSP phosphorylation, remains unknown. (2) Methods: The ability of three known SR protein kinases (SRPK1, CLK1, and AKT2) to phosphorylate the RBM20 RSRSP stretch and regulate target gene splicing was evaluated by using both in vitro and in vivo approaches. (3) Results: We found that all three kinases phosphorylated S638 and S640 in the RSRSP stretch and regulated RBM20 target gene splicing. While SRPK1 and CLK1 were both capable of directly phosphorylating the RS domain in RBM20, whether AKT2-mediated control of the RS domain phosphorylation is direct or indirect could not be determined. (4) Conclusions: Our results indicate that SR protein kinases regulate the splicing of a cardiomyopathy-relevant gene by modulating phosphorylation of the RSRSP stretch in RBM20. These findings suggest that SR protein kinases may be potential targets for the treatment of RBM20 cardiomyopathy. [ABSTRACT FROM AUTHOR]

Leydig cells (LCs) are the predominant cells of androgen production, which plays key roles in spermatogenesis and maintaining male secondary sexual characteristics. Abnormal development of LCs affects androgen levels in vivo, affects fertility and may even lead to infertility. Little is known about the regulation mechanism on LCs' development and maturation in domestic animals, especially the regulation of non-coding RNAs. In this study, we continued to dig deeper in the previous RNA-seq data of porcine LCs from our group, combined with detecting the expression profiles in different tissues and different types of cells in the testis, to screen out candidate microRNAs (miRNAs) that may affect the regulation of LCs. A total of two miRNAs, ssc-miR-21-5p and ssc-miR-615 ("ssc" is omitted below), were finally determined. After overexpression and interference of miRNAs in vitro, the effects of candidate miRNAs on the proliferation and apoptosis of TM3 (mouse Leydig cell line) were explored. The results showed that miR-21-5p led to a decrease in TM3 cell density and p53 (apoptosis related protein) expression. Meanwhile, miR-21-5p decreased EdU positive cell numbers, but increased TUNEL positive cell numbers, suggesting miR-21-5p could inhibit proliferation and promote apoptosis. Conversely, miR-615 could increase TM3 cell density. Western blot and TUNEL assay indicated miR-615 inhibited apoptosis, but had no effect on proliferation. In addition, Sox5 was identified a potential target gene of these two miRNAs by Dual-Luciferase reporter system assay. Our findings about functions of miRNAs in TM3 and the mapping of miRNAs-target gene regulatory network would provide an important basis for the further elucidation of miRNAs in regulating pig LCs. [ABSTRACT FROM AUTHOR]

RNA binding motif 20 (RBM20) cardiomyopathy has been detected in approximately 3% of populations afflicted with dilated cardiomyopathy (DCM). It is well conceived that RBM20 cardiomyopathy is provoked by titin isoform switching in combination with resting Ca2+ leaking. In this study, we characterized the cardiac function in Rbm20 knockout (KO) rats at 3‐, 6‐, 9‐, and 12‐months of age and examined the effect of the ryanodine receptor stabilizer S107 on resting intracellular levels and cardiomyocyte contractile properties. Our results revealed that even though Rbm20 depletion promoted expression of larger titin isoform and reduced myocardial stiffness in young rats (3 months of age), the established DCM phenotype required more time to embellish. S107 restored elevated intracellular Ca2+ to normal levels and ameliorated cardiomyocyte contractile properties in isolated cardiomyocytes from 6‐month‐old Rbm20 KO rats. However, S107 failed to preserve cardiac homeostasis in Rbm20 KO rats at 12 months of age, unexpectedly, likely due to the existence of multiple pathogenic mechanisms. Taken together, our data suggest the therapeutic promises of S107 in the management of RBM20 cardiomyopathy. [ABSTRACT FROM AUTHOR]

Cell division cycle 25A (CDC25A), a member of the CDC25 family of phosphatases, is required for progression from G1 to the S phase of the cell cycle. CDC25A provides an essential function during early embryonic development in mice, suggesting that it plays an important role in growth and development. In this study, we used mathematical expectation (ME) methods to identify a 20-bp insertion/deletion (indel) polymorphism of CDC25A gene in Shaanbei White Cashmere (SBWC) goats. We also investigated the association between this 20-bp indel and growth-related traits in SBWC goats. Association results showed that the indel was related to growth traits (height at hip cross, cannon circumference, and cannon circumference index) in SBWC goats. The height at hip cross of individuals with insertion/insertion (II) genotype was higher than those with insertion/deletion (ID) genotype (P=0.02); on the contrary, the cannon circumference and cannon circumference index of individuals with ID genotype were superior when compared with those with II genotype (P=0.017 and P=0.009). These findings suggest that the 20-bp indel in the CDC25A gene significantly affects growth-related traits, and could be utilized as a candidate marker for marker-assisted selection (MAS) in the cashmere goat industry. [ABSTRACT FROM AUTHOR]

Pre-mRNA splicing plays an important role in muscle function and diseases. The RNA binding motif 20 (RBM20) is a splicing factor that is predominantly expressed in muscle tissues and primarily regulates pre-mRNA splicing of Ttn, encoding a giant muscle protein titin that is responsible for muscle function and diseases. RBM20-mediated Ttn splicing has been mostly studied in heart muscle, but not in skeletal muscle. In this study, we investigated splicing specificity in different muscle types in Rbm20 knockout rats and hormonal effects on RBM20-mediated splicing both in cellulo and in vivo studies. The results revealed that RBM20 is differentially expressed across muscles and RBM20-mediated splicing is muscle-type specific. In the presence of RBM20, Ttn splicing responds to hormones in a muscle-type dependent manner, while in the absence of RBM20, Ttn splicing is not affected by hormones. In differentiated and undifferentiated C2C12 cells, RBM20-mediated splicing in response to hormonal effects is mainly through genomic signaling pathway. The knowledge gained from this study may help further understand muscle-specific gene splicing in response to hormone stimuli in different muscle types. [ABSTRACT FROM AUTHOR]

Simple Summary: In the field of animal breeding, selection using molecular genetic markers, such as insertions/deletions (indels), is a novel method to reduce the generation interval and more accurately improve performance. The casein alpha s1 (CSN1S1) gene is one of the major genes regulating the milk performance of mammals, and it is also relevant to body development. Previous studies have shown that an 11-bp indel in this gene was strongly associated with goat litter size, however, its effects on milk performance and body measurement traits have not been studied. In this study, this indel was genotyped in three Chinese goat breeds, and further analysis results revealed that this indel polymorphism was significantly associated with milk performance in dairy goats, as well as affecting the body measurement traits in all three breeds. These findings hint that the 11-bp indel within CSN1S1 gene could be utilized for effectively selecting goats with excellent performances in terms of production and breeding. The casein alpha s1 (CSN1S1) gene encodes α-s1 casein, one of the proteins constituting milk, which affects milk performance, as well as improving the absorption of calcium and bone development in mammals. A previous study found that an 11-bp insertion/deletion (indel) of this gene strongly affected litter size in goats. However, to our knowledge, the relationships between this polymorphism and the milk performance and body measurement traits of goats have not been reported. In this paper, the previously identified indel has been recognized in three Chinese goat breeds, namely the Guanzhong dairy goat (GZDG; n = 235), Shaanbei white cashmere goat (SBWC; n = 1092), and Hainan black goat (HNBG; n = 278), and the following three genotypes have been studied for all of the breeds: insertion/insertion (II), deletion/deletion (DD), and insertion/deletion (ID). The allele frequencies analyzed signified that the frequencies of the "D" allele were higher (47.8%–65.5%), similar to the previous report, which indicates that this polymorphism is genetically stable in different goat breeds. Further analysis showed that this indel was markedly associated with milk fat content, total solids content, solids-not-fat content, freezing point depression, and acidity in GZDG (p < 0.05), and also affected different body measurement traits in all three breeds (p < 0.05). The goats with II genotypes had superior milk performance, compared with the others; however, goats with DD genotypes had better body measurement sizes. Hence, it may be necessary to select goats with an II or DD genotype, based on the desired traits, while breeding. Our study provides information on the potential impact of the 11-bp indel polymorphism of the CSN1S1 gene for improving the milk performance and body measurement traits in goats. [ABSTRACT FROM AUTHOR]

Simple Summary: POU (Pit-Oct-Unc) class 1 homeobox 1 (POU1F1, or Pit-1) is a key transcription factor that directly regulates pituitary hormone-related genes and was considered as a candidate gene for reproduction and growth traits in goat breeding. Three important single nucleotide polymorphisms (SNPs), c.682G > T, c.723T > G and c.837T > C, were found in Shaanbei white cashmere (SBWC) goats by direct DNA sequencing, and c.876 + 110T > C was monomorphic. Further analyses showed that the c.682G > T and c.837T > C loci were associated with litter size, and c.682G > T, c.723T > G and c.837T > C strongly affected growth traits (p < 0.05). It is an interesting phenomenon for this population that individuals with the same genotypes had more offspring and that their body status was also better. Thus, we speculated that there was a positive correlation between growth and lambing ability. Besides this, female goats with H3H7 (combined from haplotype 3 and 7) diplotype (GTTT-TTTT), which was consistent with the combination of the optimal genotype of four SNPs, had a better litter size and growth status than other diplotypes. This study provides the first association analyses between the SNPs of POU1F1 and economic traits in SBWC goats, which could be considered as effective SNP markers for genomic selection in goat breeding. POU (Pit-Oct-Unc) class 1 homeobox 1 (POU1F1, or Pit-1) is a transcription factor that directly regulates pituitary hormone-related genes, as well as affects the reproduction and growth in mammals. Thus, POU1F1 gene was investigated as a candidate gene for litter size and growth performance in goats. In the current study, using direct DNA sequencing, c.682G > T, c.723T > G and c.837T > C loci were genotyped in Shaanbei white cashmere (SBWC) goats (n = 609), but c.876 + 110T > C was monomorphic. Besides, the c.682G > T locus was first identified by HinfI (Haemophilus influenzae Rf) restriction endonuclease. Association analysis results showed that the c.682G > T, c.837T > C loci and diplotypes were significantly associated with goat litter size (p < 0.05). The positive genotypes were GT and TT for the two SNPs, respectively, and the optimal diplotype was H3H7 (GTTT-TTTT). On the other hand, the c.682G > T, c.723T > G and c.837T > C strongly affected growth traits and body measurement indexes in SBWC goats (p < 0.05). The positive genotypes or allele of these SNPs were GT, G and TT, respectively. Additionally, the goats with H3H7 diplotype also had a greater growth status than others (p < 0.05). Here, individuals with same genotype had both a better litter size and growth traits, showing a positive correlation between these economic traits. Meanwhile, the positive genotypes of four SNPs were combined to obtain the optimal diplotype, which was also H3H7. These SNPs, especially the diplotype, could be used for the genomic selection of excellent individuals with a greater litter size and better growth status in goat breeding. [ABSTRACT FROM AUTHOR]