Identification of two transcript variants of the gene <i>Spata33</i>.

<p>A). Schematic representation of the genomic structure and alternative exons of the gene <i>Spata33</i>. Mouse <i>Spata33</i> was spliced by alternative 5′ splice site. White blocks represent 3 exons and black block shows alternative exon. B). Schematic view of alternatively spliced transcripts of the <i>Spata33</i> gene. Transcript a (GenBank: NM_177279.4) encodes a 132-aa protein. An insertion of expanded exon 2 results in a frameshift, which causes early termination of transcript b (GenBank: CA466259.1). Numbers above the wide rectangles show the amino acid sites, and the numbers below represent nucleotide sites. The locations of the stop codons for each variant are showed in white triangles. Arrows with P1S and P1A indicate RT-PCR primers, and arrows with P2S and P2A denote quantitative RT-PCR primers.</p

Experimental and influencing factors of corn stalk pulling force

ABSTRACT The harvesting of straw by the flail knife type straw cutting device will cause loose contacts between straw roots and soil, affecting the straw feed's impurity content. In this study, a theoretical analysis of the straw cutting process was conducted to explore the factors influencing the root-soil disturbance. A pulling force test device was designed to test the pulling force of the corn stalk. The response surface method was used to study the effects of various factors on the straw pulling force under different conditions of draft angles (20°, 30°, 40°), soil moisture content (15.23%, 17.62%, 20.47%), and different straw root diameters. The test results showed that the straw pulling force was directly proportional to the straw root diameter and inversely proportional to the soil moisture content. The straw pulling force decreased first and then increased with the increase of the draft angle. According to the established second-order regression model, when the draft angle was 28.5°, the soil moisture content was 20.47%, the root diameter was 22 mm, the minimum pulling force of straw was 189.635N. The test results can provide a reference for the design of straw feed harvester.</div

Phylogenetic tree, amino acid alignments, domains and modification sites of Spata33.

<p>A). Phylogenetic tree of Spata33 in mammals. Phylogenetic analysis was performed with Phylip. Numbers on the branches represent the bootstrap values from 1000 replicates obtained using the Neighbor-Joining method. The scale bar corresponds to the estimated evolutionary distance units. GenBank accession numbers are as follows: <i>Callithrix jacchus</i>, XP_002761326.1; <i>Canis lupus familiaris</i>, XP_003434741.1; <i>Cricetulus griseus</i>, XP_003495126.1; <i>Gorilla gorilla gorilla</i>, XP_004058209.1; <i>Homo sapiens,</i> BAG64150.1; <i>Macaca mulatta,</i> XP_001104069.2; <i>Mus musculus</i>, NP_796253.2; <i>Nomascus leucogenys</i>, XP_003280677.1; <i>Otolemur garnettii,</i> XP_003800884.1; <i>Pan paniscus</i>, XP_003805830.1; <i>Pan troglodytes</i>, XP_511172.4; <i>Papio anubis,</i> P_003917381.1; <i>Rattus norvegicus</i>, NP_001099665.1; <i>Saimiri boliviensis boliviensis</i>, XP_003944616.1; <i>Tupaia_chinensis</i>, ELW62868.1. B). Alignment of amino acid sequences of the Spata33 proteins. Amino acids that are identical in all these species are shown in white letters on black background. The DUF4609 domain predicted by Pfam is boxed. C). Schematic mapping of potential protein domains and post-translational modification sties. The predicted sites for N-myristoylation, N-glycosylation and phosphorylations in Spata33 were indicated (N-Myr, N-myristoylation site; N-Glyc, N-glycosylation site; S/T, Serine/Threonine phosphorylation sites).</p

Immunostaining of Spata33 protein in the juvenile (aged 10–20 days) and mature testes (aged 35 days) using anti-Spata33 antibody (A), and subcellular localization of Spata33 protein in the GC-1 cells and TM4 cells (B).

<p>A). Spata33 was expressed mainly in the spermatogonia (white arrow heads), spermatocytes (red arrows) and round spermatids (red arrow heads). The signals were observed in both the cytosol and nuclei from P12-P35, whereas no signals were detected in P10. In controls, no positive signals were observed in P35 testis sections when Spata33 antibody was replaced by 1% normal rabbit serum. Nuclei were re-dyed with Hematoxylin (blue). Bars, 20 µm. B). GC-1 and TM4 cells were transfected with Spata33-GFP (green, Excitation 488 nm, Emission 507 nm) and stained with Hoechst (blue, Excitation 352 nm, Emission 461 nm) respectively. Spata33 is localized both in the nuclei and cytoplasm within GC-1 cells or TM4 cells. Bar, 10 µm.</p

Data_Sheet_1_Case report: Whole-exome sequencing identifies a novel DES mutation (p. E434K) in a Chinese family with cardiomyopathy and sudden cardiac death.xls

BackgroundDesmin is an intermediate filament protein that plays a critical role in the stabilization of the sarcomeres and cell contacts in the cardiac intercalated disk. Mutated DES gene can cause hereditary cardiomyopathy with heterogeneous phenotypes, while the underlying molecular mechanisms requires further investigation.MethodsWe described a Chinese family present with cardiomyopathy and sudden cardiac death (SCD). Whole-exome sequencing (WES) and bioinformatics strategies were employed to explore the genetic entity of this family.ResultsAn unknown heterozygote missense variant (c.1300G > A; p. E434K) of DES gene was identified. The mutation cosegregates in this family. The mutation was predicted as pathogenic and was absent in our 200 healthy controls.ConclusionWe identified a novel DES mutation (p. E434K) in a Chinese family with cardiomyopathy and SCD. Our study not only provided a new case for the study of the relationship between DES mutations and hereditary cardiomyopathy but also broadened the spectrum of DES mutations.</p

Expression of <i>Spata33</i> mRNA and protein in the postnatal testes of mice.

<p>A). RT-PCR showed expression pattern of <i>Spata33</i> mRNA in testes from P2 to P20 (postnatal days). Total RNAs were isolated from mouse testes and then cDNAs were synthesized. <i>Hprt</i> was used as an internal control. Molecular weight is shown on the right. <i>Spata33</i> mRNA was increased markedly after P12. B). Real-time fluorescent quantitative PCR of the <i>Spata33</i> in testes from P2 to P20. Error bars indicate the standard deviation (SD) of the mean (n = 3). Y-axis represents relative expression levels of <i>Spata33</i> and X-axis shows different development stages P2-P20. C). Expression of Spata33 protein in testes from P2 to P20. Mouse testes were individually collected from aged 2 to 20 days and were subjected to Western blot analysis with antibody against Spata33. The protein level was markedly increased from P12. Actin was used as an internal control. Molecular weight is shown on the right. All experiments were performed three times with independent individuals.</p

Expression pattern of <i>Spata33</i> in various tissues of adult mouse.

<p>A). RT-PCR of <i>Spata33</i> in adult tissues. <i>Hprt</i> was used as a standard. The gene <i>Spata33</i> was predominantly expressed in testis. Molecular weight is shown on the right. B). Real-time fluorescent quantitative PCR of the <i>Spata33</i> gene in adult tissues. Error bars indicate the standard deviation (SD) of the mean (n = 3). Y-axis represents relative expression levels of <i>Spata33</i> and X-axis shows different mouse tissues. C). Expression of Spata33 protein in various tissues. Mouse tissues were subjected to Western blot analysis with antibody against Spata33. Spata33 protein recognized a band at 15 KD. The Spata33 protein was predominantly expressed in testis. Actin was used as an internal control. Molecular weight is shown on the right. We repeated each experiment three times with independent individuals.</p

Image_1_Case report: Whole-exome sequencing identifies a novel DES mutation (p. E434K) in a Chinese family with cardiomyopathy and sudden cardiac death.PDF

BackgroundDesmin is an intermediate filament protein that plays a critical role in the stabilization of the sarcomeres and cell contacts in the cardiac intercalated disk. Mutated DES gene can cause hereditary cardiomyopathy with heterogeneous phenotypes, while the underlying molecular mechanisms requires further investigation.MethodsWe described a Chinese family present with cardiomyopathy and sudden cardiac death (SCD). Whole-exome sequencing (WES) and bioinformatics strategies were employed to explore the genetic entity of this family.ResultsAn unknown heterozygote missense variant (c.1300G > A; p. E434K) of DES gene was identified. The mutation cosegregates in this family. The mutation was predicted as pathogenic and was absent in our 200 healthy controls.ConclusionWe identified a novel DES mutation (p. E434K) in a Chinese family with cardiomyopathy and SCD. Our study not only provided a new case for the study of the relationship between DES mutations and hereditary cardiomyopathy but also broadened the spectrum of DES mutations.</p

Investigation of Supercritical Methane Adsorption of Overmature Shale in Wufeng-Longmaxi Formation, Southern Sichuan Basin, China

Accurately determining the gas sorption capacity of a specific shale reservoir is critical for further assessment of shale gas reserves. A series of high-pressure methane adsorption measurements were conducted at 60 °C with a pressure of up to 30 MPa for Wufeng-Longmaxi shales from the southern Sichuan Basin, which is considered as the most promising shale-gas target in China, to evaluate the fitting quality of different excess adsorption models and to determine the effect of organic matter content, maturity, mineralogy, and pore structure on the gas adsorption capacity. Both the Langmuir- and supercritical Dubinin–Radushkevich (SDR)-based adsorption models are closely fitted with the measured excess adsorption amount. However, the freely fitted SDR model is considered to be the most reasonable model, in which the adsorbed-phase density is always lower than the liquid methane density at the boiling point (0.424 g/cm3) and the average relative error (ARE) is relatively small. Adsorbed-phase density is a key parameter for calculating absolute adsorption isotherms. For a specific shale, a lower constant adsorbed-phase density applied in the adsorption model would result in higher absolute adsorption capacity. For the Langmuir-based model, the actual absolute adsorption capacity would be underestimated when adsorption experiments were only conducted at the low-pressure range (0–15 MPa). The methane adsorption capacities show a great positive correlation with the total organic carbon (TOC) content. The TOC-normalized adsorption capacities have a negative relationship with maturity at an overmature stage. The clay content shows a positive correlation with the TOC-normalized adsorption capacities, indicating that clays also make some contribution to methane sorption on these organic-rich shales. Furthermore, methane adsorption in overmature shales is mainly controlled by the structure of the pore <20 nm in size, revealing that the adsorbed methane is occupied not only in micropores but also in fine mesopores

Table_1_Case Report: Identification of the First Synonymous Variant of Myosin Binding Protein C3 (c.24A>C, p.P8P) Altering RNA Splicing in a Cardiomyopathy and Sudden Cardiac Death Case.docx

BackgroundSudden cardiac death (SCD), based on sudden cardiac ejection cessation, is an unexpected death. Primary cardiomyopathies, including dilated cardiomyopathy (DCM), are one of main causes of SCD. The DCM is characterized by a cardiac dilatation and a reduced systolic function with a prevalence of 1/250 in adults. The DCM has been reported with more than 60 disease-causing genes, and MYBPC3 variants are one of the most common and well-known causes of DCM.MethodsWe identified a 29-year-old female who died of SCD. We performed a whole-exome sequencing (WES) to detect her genetic etiology and used minigene modeling and immunohistochemistry staining to verify the pathogenicity.ResultsWe determined that the woman died of SCD caused by DCM due to an identified novel synonymous variant of MYBPC3 (NM_000256.3: c.24A>C, p.P8P) in the deceased. The variant can result in abnormal splicing, which was confirmed by minigene models and immunohistochemistry staining.ConclusionWe may have identified the first deleterious synonymous variant of MYBPC3 in an SCD case and verified its significant impact on RNA splicing. Our description enriched the spectrum of MYBPC3 variants and emphasized the significance of synonymous variants that are always disregarded in genetic screening.</p