Deficiency of CAMSAP2 impairs olfaction and the morphogenesis of mitral cells.

In developing olfactory bulb (OB), mitral cells (MCs) remodel their dendrites to establish the precise olfactory circuit, and these circuits are critical for individuals to sense odors and elicit behaviors for survival. However, how microtubules (MTs) participate in the process of dendritic remodeling remains elusive. Here, we reveal that calmodulin-regulated spectrin-associated proteins (CAMSAPs), a family of proteins that bind to the minus-end of the noncentrosomal MTs, play a crucial part in the development of MC dendrites. We observed that Camsap2 knockout (KO) males are infertile while the reproductive tract is normal. Further study showed that the infertility was due to the severe defects of mating behavior in male mice. Besides, mice with loss-of-function displayed defects in the sense of smell. Furthermore, we found that the deficiency of CAMSAP2 impairs the classical morphology of MCs, and the CAMSAP2-dependent dendritic remodeling process is responsible for this defect. Thus, our findings demonstrate that CAMSAP2 plays a vital role in regulating the development of MCs.

CAMSAP1 role in orchestrating structure and dynamics of manchette microtubule minus-ends impacts male fertility during spermiogenesis.

The manchette is a crucial transient structure involved in sperm development, with its composition and regulation still not fully understood. This study focused on investigating the roles of CAMSAP1 and CAMSAP2, microtubule (MT) minus-end binding proteins, in regulating manchette MTs, spermiogenesis, and male fertility. The loss of CAMSAP1, but not CAMSAP2, disrupts the well-orchestrated process of spermiogenesis, leading to abnormal manchette elongation and delayed removal, resulting in deformed sperm nuclei and tails resembling oligoasthenozoospermia symptoms. We investigated the underlying molecular mechanisms by purifying manchette assemblies and comparing them through proteomic analysis, and results showed that the absence of CAMSAP1 disrupted the proper localization of key proteins (CEP170 and KIF2A) at the manchette minus end, compromising its structural integrity and hindering MT depolymerization. These findings highlight the significance of maintaining homeostasis in manchette MT minus-ends for shaping manchette morphology during late spermiogenesis, offering insights into the molecular mechanisms underlying infertility and sperm abnormalities.

Ferroptosis-Related lncRNA to Predict the Clinical Outcomes and Molecular Characteristics of Kidney Renal Papillary Cell Carcinoma.

Kidney renal papillary cell carcinoma (KIRP) is a highly heterogeneous type of kidney cancer, resulting in limited effective prognostic targets for KIRP patients. Long non-coding RNAs (lncRNAs) have emerged as crucial regulators in the regulation of ferroptosis and iron metabolism, making them potential targets for the treatment and prognosis of KIRP. In this study, we constructed a ferroptosis-related lncRNA risk score model (FRM) based on the TCGA-KIRP dataset, which represents a novel subtype of KIRP not previously reported. The model demonstrated promising diagnostic accuracy and holds potential for clinical translation. We observed significant differences in metabolic activities, immune microenvironment, mutation landscape, ferroptosis sensitivity, and drug sensitivity between different risk groups. The high-risk groups exhibit significantly higher fractions of cancer-associated fibroblasts (CAFs), hematopoietic stem cells (HSC), and pericytes. Drugs (IC50) analysis provided a range of medication options based on different FRM typing. Additionally, we employed single-cell transcriptomics to further analyze the impact of immune invasion on the occurrence and development of KIRP. Overall, we have developed an accurate prognostic model based on the expression patterns of ferroptosis-related lncRNAs for KIRP. This model has the potential to contribute to the evaluation of patient prognosis, molecular characteristics, and treatment modalities, and can be further translated into clinical applications.

A Tailorable Series of Elastomeric-To-Rigid, Selfhealable, Shape Memory Bismaleimide.

In recent years, there has been rapid development in the field of shape memory materials with active deformation performance. However, bismaleimide, a widely used thermosetting material in aerospace, has been largely overlooked in shape memory applications. This work presents the synthesis of a molecule containing an alkene bond adjacent to an oxygen atom. Through molecular design, a one-time reaction between this specialized molecule and the bismaleimide molecule is successfully achieved, facilitated by the steric hindrance effect. Therefore, a new series of shape memory bismaleimide materials are obtained. By introducing a diamine to adjust the chain length, the properties of material are further improved, resulting in increasing static modulus by 506 times. The synthesized materials exhibit a broad glass transition temperature (Tg) range exceeding 153 °C, remarkable stiffness tunability. Notably, in the synthesis process of this materials series, the disulfide bonds are introduced, which facilitates the realization of self-healing and reprocessable functionalities in the resulting thermosetting materials. This significant advancement lays a solid foundation for the future recycling and reuse of aircraft, satellites, and other equipment, offering promising prospects for enhancing sustainability and efficiency within the aerospace industry.

Lateral shearing interferometry method based on double-checkerboard grating by suppressing aliasing effect.

Ronchi lateral shearing interferometry is a promising wavefront sensing technology with the advantages of simple structure and no reference light, which can realize a high-precision wavefront aberration measurement. To obtain shear information in both directions, the conventional double-Ronchi interferometer sequentially applies two orthogonal one-dimensional Ronchi gratings as the object-plane splitting element of the optics under test. Simultaneously, another Ronchi grating is positioned on the image plane in the same orientation to capture two sets of interferograms, thereby enabling two-dimensional wavefront reconstruction. Mechanical errors will inevitably be introduced during grating conversion, affecting reconstruction accuracy. Based on this, we propose a lateral shearing interferometry applying double-checkerboard grating. Only unidirectional phase shift is needed to obtain shear information in two directions while evading the grating conversion step, aiming to streamline operational processes and mitigate the potential for avoidable errors. We employ scalar diffraction theory to analyze the full optical path propagation process of the double-checkerboard shearing interferometry and introduce a new reconstruction algorithm to effectively extract the two-dimensional shear phase by changing the grating morphology, suppressing the aliasing effect of irrelevant diffraction orders. We reduce the fitting error through iterative optimization to realize high-precision wavefront reconstruction. Compared with conventional Ronchi lateral shearing interferometry, the proposed method exhibits better robustness and stability in noisy environments.

Fast wavefront sensing method based on diffraction basis vectors for tightly focused optical systems.

Recently, phase retrieval techniques have garnered significant attention with their exceptional flexibility. However, their application is limited in optical systems with high numerical aperture due to the disregarded polarization properties of the beam. In this paper, a fast wavefront sensing method for tightly focused systems is proposed. Firstly, a vector diffraction model based on the chirp-Z transform is established to analytically describe the focal spot using the modal coefficients of polynomials and diffraction basis vectors, which accommodating any pixel size and resolution, thereby enabling to break through sampling constraints and remove lateral errors. Additionally, a modified Newton-gradient second-order algorithm is introduced to simultaneously optimize wavefront in multiple polarization directions, without the need for diffraction operators during iterations. Both numerical simulations and error analysis confirm the efficacy and precision of the proposed wavefront sensing method.

Design of a panoramic annular lens system with an ultra-wide angle via an annular Gaussian radial basis function freeform surface.

The panoramic annular lens (PAL) system can capture plentiful scene information in real time. The locally described freeform surface can provide more degrees of freedom for the optical system design than the globally described one. In this paper, we propose a locally described annular Gaussian radial basis function freeform surface that enables a high-performance yet compact PAL system. The designed PAL system achieves an ultra-wide field of view (FOV) of (30∘∼125∘)×360∘. The F-theta distortion of the system is less than 3%, and the modulation transfer function across the whole FOV is greater than 0.5 at 100 lp/mm. The proposed system can be implemented for various applications such as video conferencing or robotic navigation.

CAMSAP1 breaks the homeostatic microtubule network to instruct neuronal polarity.

The establishment of axon/dendrite polarity is fundamental for neurons to integrate into functional circuits, and this process is critically dependent on microtubules (MTs). In the early stages of the establishment process, MTs in axons change dramatically with the morphological building of neurons; however, how the MT network changes are triggered is unclear. Here we show that CAMSAP1 plays a decisive role in the neuronal axon identification process by regulating the number of MTs. Neurons lacking CAMSAP1 form a multiple axon phenotype in vitro, while the multipolar-bipolar transition and radial migration are blocked in vivo. We demonstrate that the polarity regulator MARK2 kinase phosphorylates CAMSAP1 and affects its ability to bind to MTs, which in turn changes the protection of MT minus-ends and also triggers asymmetric distribution of MTs. Our results indicate that the polarized MT network in neurons is a decisive factor in establishing axon/dendritic polarity and is initially triggered by polarized signals.

First pilot trial of colorectal ESD guided by a new magnetic anchor for ease of placement.

BACKGROUND: In recent years, studies have demonstrated that magnetic anchor-guided endoscopic submucosal dissection (MAG-ESD) is feasible and safe and may facilitate the treatment of all difficult lesions. However, the major problem with MAG-ESD is the inability to deliver the magnetic anchor to the gastrointestinal tract without withdrawal or reinsertion of the endoscope. Therefore, our team developed a magnetic anchor that could be easily inserted through the biopsy channel, facilitating ESD traction and evaluated its effectiveness and safety. METHODS: The study was conducted between October 2020 and June 2021 at The Second Affiliated Hospital of Harbin Medical University, China. One hundred and twelve patients with colorectal tumors treated with ESD were divided into two groups for historical control comparison. A channel-placed magnetic anchor (CPMAG) group and a control group consisting of patients who had conventional ESD without adjuvant traction. The rate of en bloc resection and resection with tumor-free lateral/basal margins (R0 resection), dissection speeds, procedure time, intraoperative bleeding and perforation complications, and postoperative follow-up were compared between the two groups, so as to evaluate the clinical effect and safety of the new magnetic anchor. RESULTS: The en bloc resection and R0 resection rate with CPMAG-ESD were slightly higher than with conventional ESD but this was not statistically significant. The median dissection speeds with CPMAG-ESD were higher than with conventional ESD, but the difference was not statistically significant. Intraoperative bleeding and postoperative complications with the CPMAG-ESD were less than with conventional ESD, but this was not statistically significant. The median operating time was shorter with CPMAG- ESD than with conventional ESD (24.5 min [range 15.8-66.5 min] vs 39 min [range 29-58 min], p = 0.024), and this difference was statistically significant. CONCLUSIONS: The new magnetic anchor-guided ESD technique appears to be a feasible and safe method for treating early colorectal tumors with en bloc resection, with improvement of the submucosal visual field, and less adverse events.

The role of EMILIN-1 in the osteo/odontogenic differentiation of dental pulp stem cells.

BACKGROUND: Human dental pulp stem cells (hDPSCs) may be the best choice for self-repair and regeneration of teeth and maxillofacial bone tissue due to their homogeneous tissue origin, high proliferation and differentiation rates, and no obvious ethical restrictions. Recently, several studies have shown that extracellular matrix (ECM) proteins can effectively regulate the proliferation and differentiation fate of mesenchymal stem cells (MSCs). However, the role of elastin microfibril interface-located protein-1 (EMILIN-1), a new ECM glycoprotein, in osteo/odontogenic differentiation of hDPSCs has not been reported. The aim of this study was to explore the effect of EMILIN-1 during osteo/odontogenic differentiation of hDPSCs. METHODS: hDPSCs were cultured in osteo/odontogenic induction medium. qPCR and Western blot analysis were performed to detect osteo/odonto-specific genes/proteins expression as well as the expression of EMILIN-1. After knockdown of Emilin-1 in hDPSCs with small interfering RNA and exogenous addition of recombinant human EMILIN-1 protein (rhEMILIN-1), Cell Counting Kit-8 assay, alkaline phosphatase staining, alizarin red S staining, qPCR and Western blot were performed to examine the effect of EMILIN-1 on proliferation and osteo/odontogenic differentiation of hDPSCs. RESULTS: During the osteo/odontogenic induction of hDPSCs, the expression of osteo/odonto-specific genes/proteins increased, as did EMILIN-1 protein levels. More notably, knockdown of Emilin-1 decreased hDPSCs proliferation and osteo/odontogenic differentiation, whereas exogenous addition of rhEMILIN-1 increased them. CONCLUSIONS: These findings suggested that EMILIN-1 is essential for the osteo/odontogenic differentiation of hDPSCs, which may provide new insights for teeth and bone tissue regeneration.

Semi-analytic Fresnel diffraction calculation with polynomial decomposition.

The numerical method based on the fast Fourier transform (FFT) is generally applied to calculate the Fresnel diffraction field, which would suffer from sampling constraints. To break this limit, in this Letter, the semi-analytic Fresnel diffraction calculation method is proposed based on polynomial decomposition. The diffraction field is computed by using properly analytic Fresnel diffraction basis functions (FDBFs) according to the application requirements. Analytic FDBF is calculated based on Legendre or Chebyshev polynomials by using the object-domain frequency division multiplexing method. The proposed method offers arbitrary sampling, high-flexibility, and high-accuracy diffraction calculation in the full Fresnel region. The computational efficiency and accuracy of the proposed method are compared with FFT-based methods. It has potential application in light field analysis, wavefront sensing, and image processing.

Occlusion culling for computer-generated cylindrical holograms based on a horizontal optical-path-limit function.

Recently, great progress has been made in the research of cylindrical holography as a promising technique of 360° display. However, there is an unsolved issue of occlusion culling, which is critical to cylindrical holography and degrades the reconstructed images due to overlapping. To our knowledge, the occlusion issue in cylindrical holography has never been deeply discussed. In this paper, a method of occlusion culling is proposed for computer-generated cylindrical holograms based on a horizontal optical-path-limit function. In cylindrical diffraction, the propagation characteristics of light waves can be described by the point spread function, which is mainly obtained by analyzing the meaning of the obliquity factor in the concentric cylinder model. Different from the planar diffraction, the diffraction area of each source point is limited within the tangents in cylindrical diffraction. Therefore, a horizontal optical path limit function that acts directly on the point spread function for occlusion culling is established. Besides, the proposed method can be applied to the three-dimensional object by using the layer-oriented method. Moreover, the effectiveness of the proposed occlusion culling method is verified by the numerical simulation results and error analysis of the reconstructed images.

Spherical self-diffraction for speckle suppression of a spherical phase-only hologram.

The spherical computer-generated hologram is inevitably suffered from the speckle noise since it is necessary to add random phase to the object to ensure the scattering characteristic of reconstructed image. The speckle noise seriously degrades the quality of reconstructed image especially for a spherical phase-only hologram (SPOH). In this paper, spherical self-diffraction iteration (SSDI) algorithm is proposed to suppress the speckle noise in the SPOH. The algorithm is based on spherical self-diffraction (SSD) model which is a special case of spherical back-propagation (SBP) model at limit condition. The correctness of SBP and SSD as well as the effectiveness of SSDI algorithm are verified by numerical simulations. Meaningfully, the proposed method significantly outperforms the conventional methods in speckle suppression performance and computing speed. As far as we known, models of SBP and SSD as well as conception of SSDI are firstly proposed and applied for speckle suppression of SPOH.

Genome-Wide Analysis of CCA1-Like Proteins in Soybean and Functional Characterization of GmMYB138a.

Plant CIRCADIAN CLOCK ASSOCIATED1 (CCA1)-like proteins are a class of single-repeat MYELOBLASTOSIS ONCOGENE (MYB) transcription factors generally featured by a highly conserved motif SHAQK(Y/F)F, which play important roles in multiple biological processes. Soybean is an important grain legume for seed protein and edible vegetable oil. However, essential understandings regarding CCA1-like proteins are very limited in soybean. In this study, 54 CCA1-like proteins were identified by data mining of soybean genome. Phylogenetic analysis indicated that soybean CCA1-like subfamily showed evolutionary conservation and diversification. These CCA1-like genes displayed tissue-specific expression patterns, and analysis of genomic organization and evolution revealed 23 duplicated gene pairs. Among them, GmMYB138a was chosen for further investigation. Our protein-protein interaction studies revealed that GmMYB138a, but not its alternatively spliced isoform, interacts with a 14-3-3 protein (GmSGF14l). Although GmMYB138a was predominately localized in nucleus, the resulting complex of GmMYB138a and GmSGF14l was almost evenly distributed in nucleus and cytoplasm, supporting that 14-3-3s interact with their clients to alter their subcellular localization. Additionally, qPCR analysis suggested that GmMYB138a and GmSGF14l synergistically or antagonistically respond to drought, cold and salt stresses. Our findings will contribute to future research in regard to functions of soybean CCA1-like subfamily, especially regulatory mechanisms of GmMYB138a in response to abiotic stresses.

Isolation of Human Adipose-Derived Stromal/Stem Cells from Lipoaspirates.

Adipose tissue is an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Adipose-derived stromal/stem cells (ASCs) have been widely used in tissue engineering and cell therapy. In addition, the clinical application of ASCs in the treatment of inflammation and injury has been proven a success. Here, we describe methods from our own laboratory and the literature for the isolation and expansion of Adipose-derived stromal/stem cells (ASCs). We present a large-scale procedure suitable for processing >100 mL volumes of lipoaspirate tissue specimens by collagenase digestion, a related procedure suitable for processing adipose tissue aspirates without digestion, and a procedure suitable for intact human adipose tissue, such as buccal fat pads in the maxillofacial region.

Isolation of Murine Adipose-Derived Stromal/Stem Cells Using an Explant Culture Method.

Adipose tissue provides a valuable cell source for tissue engineering, regenerative medicine, and adipose tissue biology studies. The most widely used adipose-derived stromal/stem cells (ASCs) isolation protocol involves enzymatic digestion with collagenase. However, the yield of the method often proves to be poor if not impossible for collection of sufficient stromal vascular fraction (SVF) for expansion when the sample size is small, for instance when only newborn mice are available for cell culture. Here, we describe an efficient protocol for the isolation and expansion of ASCs using explant culture as an alternative. Briefly, adipose tissue was minced after removing excess liquid. Then, the minced tissue was placed in culture dishes or flasks. The cells will migrate out of tissue and adhere to the culture surface after one or more days.

Unraveling the mechanism in l-Caldesmon regulating the osteogenic differentiation of PDLSCs: An innovative perspective.

Maxillofacial bone defect is one of the common symptoms in maxillofacial, which affects the function and aesthetics of maxillofacial region. Periodontal ligament stem cells (PDLSCs) are extensively used in bone tissue engineering. The mechanism that regulates the osteogenic differentiation of PDLSCs remains not fully elucidated. Previous studies demonstrated that l-Caldesmon (l-CALD, or CALD1) might be involved in the osteogenic differentiation of PDLSCs. Here, the mechanism by which CALD1 regulates the osteogenic differentiation of PDLSCs is investigated. The osteogenic differentiation of PDLSCs is enhanced with Cald1 knockdown. Whole transcriptome sequencing (RNA-seq) analysis shows that bone morphogenetic proteins (BMP) signaling pathway and Wingless type (Wnt) pathway have significant change with Cald1 knockdown, and the expressions of Wnt-induced secreted protein 1 (WISP1), BMP2, Smad1/5/9, and p-Smad1/5/9 are significantly upregulated, while Glycogen synthase kinase 3ß (GSK3ß) and p-GSK3ß are downregulated. In addition, subcutaneous implantation in nude mice shows that knockdown of Cald1 enhances the osteogenic differentiation of PDLSCs in vivo. Taken together, this study demonstrates that knockdown of Cald1 enhances the osteogenic differentiation of PDLSCs by BMP and Wnt signaling pathways, and provides a novel approach for subsequent clinical treatment.

Zinc Improves Semen Parameters in High-Fat Diet-Induced Male Rats by Regulating the Expression of LncRNA in Testis Tissue.

This study aimed to identify differentially expressed LncRNAs in testis tissue of male rats induced by high-fat diet and their changes after zinc supplementation, by constructing a high-fat feeding rat model, and then supplemented with zinc, and observed the expression of LncRNA in three groups of normal, high-fat fed, and zinc-intervened rats. Experimental studies show that the semen parameters of male rats with high-fat diet were decreased but recovered after zinc supplementation, and the related LncRNA also changed. Zinc may improve the high-fat diet-induced reduction of semen parameters by changing the expression of related LncRNA.

Zinc-Deficient Diet Causes Imbalance in Zinc Homeostasis and Impaired Autophagy and Impairs Semen Quality in Mice.

Zinc (Zn) is an essential trace element for human growth and its deficiency causes huge health impacts. The present study was conducted to examine the mechanisms by which Zn-deficient diet impairs reproductive function and its reversibility. Hence, SPF grade male Kunming (KM) mice were divided into three groups. Zn-normal diet group (ZN group) was provided with Zn-normal diet (Zn content = 30 mg/kg, DY19410Y) for 8 weeks. Zn-deficient diet group (ZD group) was provided with Zn-deficient diet (Zn content < 1 mg/kg, DY19401) for 8 weeks. Zn-deficient and Zn-normal diet group (ZDN group) was provided with 4 weeks Zn-deficient diet followed by 4 weeks Zn-normal diet. After 8 weeks, the overnight-fasted mice were sacrificed, and blood and organs were collected for further analysis. The results showed that Zn-deficient diet caused testicular structural disorders, decreased semen quality, imbalance in zinc homeostasis, and impaired autophagy. Semen quality, testosterone, serum Zn, testicular tissue Zn, testicular free Zn ions, alkaline phosphatase (ALP), zinc transporter 7(ZnT7), Beclin1, autophagy-related 5(ATG5), and the ratio of light chain 3(LC3) II/LC3I were significantly decreased, and ZnT4, Zrt-, Irt-like protein7 (ZIP7), and ZIP13 expression were significantly increased in ZD group mice, while the changes in above indicators caused by Zn-deficient diet were significantly alleviated in the ZDN group. It was concluded that Zn-deficient diet causes testicular structural disorders and decreased semen quality by causing imbalances in Zn homeostasis and impaired autophagy in male mice. Reproductive damages caused by Zn-deficient diet are reversible, and Zn-normal diet can alleviate them.

Mechanisms of damage to sperm structure in mice on the zinc-deficient diet.

BACKGROUND: Zinc (Zn)is an essential trace element for spermatogenesis and its deficiency causes abnormal spermatogenesis. OBJECTIVE: The present study was conducted to examine the mechanisms by which Zn-deficient diet impairs sperm morphology and its reversibility. METHODS: 30 SPF grade male Kunming (KM) mice were randomly divided into three groups, 10 mice per group. Zn-normal diet group (ZN group) was given Zn-normal diet（Zn content= 30 mg/kg）for 8 weeks. Zn-deficienct diet group (ZD group) was given Zn-deficienct diet（Zn content< 1 mg/kg）for 8 weeks. Zn-deficient and Zn-normal diet group（ZDN group）was given 4 weeks Zn-deficienct diet followed by 4 weeks Zn-normal diet. After 8 weeks, the overnight fasted mice were sacrificed, and blood and organs were collected for further analysis. RESULTS: The experimental results showed that Zn-deficienct diet leads to increased abnormal morphology sperm and testicular oxidative stress.The rate of abnormal morphology sperm, chromomycin A3(CMA3), DNA fragmentation index (DFI), malondialdehyde (MDA) were significantly increased, and a-kinase anchor protein 4(AKAP4), dynein axonemal heavy chain 1(DNAH1), sperm associated antigen 6(SPAG6), cilia and flagella associated protein 44(CFAP44), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), nuclear factor erythroid 2-related factor (NRF2), NAD(P)H:quinone oxidoreductase 1(NQO1)and heme oxygenase 1(HO1) were significantly decreased in the ZD group mice. While the changes in above indicators caused by Zn-deficient diet were significantly alleviated in the ZDN group. CONCLUSION: It was concluded that Zn-deficient diet causes abnormal morphology sperm and testicular oxidative stress in male mice. Abnormal morphology sperm caused by Zn-deficient diet are reversible, and Zn-normal diet can alleviate them.