GATA4 binding to the Sox9 enhancer mXYSRa/Enh13 is critical for testis differentiation in mouse.

In mammals, SOX9/Sox9 expression in embryonic gonads is essential for male gonadal sex determination. Multiple enhancers of Sox9 have been identified, of which the mXYSRa/Enh13 enhancer plays a crucial role in mice. SOX9 and SRY binding sites within the enhancer have been identified as functional.

Estrogenic-like compounds severely disturb germ cell formation in Japanese quail.

During avian germ cell formation, primordial germ cells (PGCs) differentiate into prospermatogonia in testicular seminiferous tubules or into oogonia in the ovarian cortex in late-stage embryos. Although estrogenic endocrine-disrupting chemicals (EDCs) have been suggested to affect the differential fate of avian germ cells, there is currently no established method to examine the effects of EDCs on the differentiation potential of germline cells due to large amount of unidentified proteins present in avian germ cells. Regarding reliable molecular probes for the detection of germ cells that differentiated from the PGCs of Japanese quail, the prospermatogonium and oogonium, respectively, integrin beta1 (ITGB1), insulin-like growth factor 2-binding protein 1 (IGF2BP1), and stimulated by retinoic acid 8 (STRA8) were identified as marker proteins by RNA-seq and liquid chromatography tandem mass spectrometry analyses.

Mid- to long-term clinical and radiological assessment of a short, titanium, porous plasma-splayed flat-tapered cementless femoral stem: An 8- to 12-year follow-up study.

Introduction: Short cementless stems offer advantages such as bone-stock preservation and minimally invasive insertion. Despite encouraging short- to midterm outcomes, the long-term clinical and radiological implications of short, titanium, and porous plasma-sprayed flat-tapered stems remain unclear.

Materials And Methods: A retrospective review of 138 primary total hip arthroplasties, conducted between 2010 and 2015 in 130 patients, with a minimum 8-year follow-up was performed.

Stereoselective Synthesis of Spirolactone Analogues of Pyrrolomorpholine Alkaloids.

The synthesis of a spirolactone analogue of xylapyrraside B, a potent antioxidant agent, is described. The key step is the stereoselective formation of the spirolactone skeleton via the formal [3+2] annulation of the isopropylidene-protected glyceraldehyde and δ-methylene lactone, mediated by trifluoroborane etherate. This study addresses the stereoselective synthesis of pyrrolomorpholine spiroketal alkaloids, enabling the production of these bioactive compounds and their analogues.

Initial formation of and sex differences in primordial germ cells in Japanese quail.

DEAD-box RNA helicase 4 (DDX4) is posited to be a key maternal germ cell factor regulating avian germ cell formation. We herein showed that the DDX4 gene product of zygotic genome activation associated with the nuclear localization of the cyclin D1 protein in presumptive primordial germ cells (PGCs) plays an essential role in the proliferation of PGCs using a CRISPR/Cas9 system approach combined with in vitro fertilization techniques in Japanese quail. A proteome analysis also revealed molecular-based differences in the features of early male and female PGCs.

Turnover of mammal sex chromosomes in the -deficient Amami spiny rat is due to male-specific upregulation of .

Mammalian sex chromosomes are highly conserved, and sex is determined by on the Y chromosome. Two exceptional rodent groups in which some species lack a Y chromosome and offer insights into how novel sex genes can arise and replace , leading to sex chromosome turnover. However, intensive study over three decades has failed to reveal the identity of novel sex genes in either of these lineages.

SOX9 and SRY binding sites on mouse mXYSRa/Enh13 enhancer redundantly regulate Sox9 expression to varying degrees.

Sox9 plays an essential role in mammalian testis formation. It has been reported that gene expression in the testes is regulated by enhancers. Among them, mXYSRa/Enh13-which is located at far upstream of the transcription start site-plays a critical role, wherein its deletion causes complete male-to-female sex reversal in mice.

Risk factors for the occurrence and protraction of patellar and patellar tendon pain in children and adolescents: a prospective cohort study of 3 years.

Background: Patellar and patellar tendon pain is a common limitation to children's participation in social and physical activities. Some factors have been implicated in the occurrence and protraction of knee pain, but the causal relationship is unknown. The purpose of this study was to investigate whether participants' physical characteristics and activity level are risk factors for the occurrence and protraction of patellar and patellar tendon pain in children and adolescents.

Foot morphology and correlation with lower extremity pain in Japanese children: A cross-sectional study of the foot posture Index-6.

Background: Abnormal foot morphology in children and adolescents is a possible risk factor for lower extremity pain. Foot posture index-6 (FPI-6) is a valid and reliable tool to assess foot morphology. However, the normative data on the age distribution for FPI-6 in Asian children and adolescents are still minimal.

Human SRY Expression at the Sex-determining Period is Insufficient to Drive Testis Development in Mice.

The sex-determining region of the Y chromosome, Sry/SRY, is an initiation factor for testis development in both humans and mice. Although the functional compatibility between murine SRY and human SRY was previously examined in transgenic mice, their equivalency remains inconclusive. Because molecular interaction and timeline of mammalian sex determination were mostly described in murine experiments, we generated a mouse model in which Sry was substituted with human SRY to verify the compatibility.

Possible role of MRI-detected osteophytes as a predictive biomarker for development of osteoarthritis of the knee: A study using data from the Osteoarthritis Initiative.

Objective: To elucidate the possible role of MRI-detected osteophytes as a predictive imaging biomarker for knee osteoarthritis (KOA).

Design: Subjects (n ​= ​303) were selected according to the following inclusion criteria from the Osteoarthritis Initiative (OAI) data set: (1) ​< ​55 years old; (2) Western Ontario and McMaster Universities Arthritis Index pain score of 0; (3) Kellgren-Lawrence (KL) system grade 0 or 1; and (4) Complete MRI data set of the right knee. A pre-OA group (POA) consisted of subjects who developed KL grade 2 or more within 96 months, and a non-OA group (NOA) that remained KL 0 or 1 during that period.

Effect of Systemic Administration of Granulocyte Colony-Stimulating Factor on a Chronic Partial-Thickness Cartilage Defect in a Rabbit Knee Joint.

Objective: Cartilage lesions in the knee joint can lead to joint mechanics changes and cause knee pain. Bone marrow stimulation (BMS) promotes cartilage regeneration by perforating the subchondral bone just below the injury and inducing bone marrow cells. This study aimed to investigate whether systemic administration of granulocyte colony-stimulating factor (G-CSF) with BMS improves repair of chronic partial-thickness cartilage defects (PTCDs).

Humanization of a tandem repeat in IG-DMR causes stochastic restoration of paternal imprinting at mouse Dlk1-Dio3 domain.

The Dlk1-Dio3 imprinted domain, regulated by an intergenic differentially methylated region (IG-DMR), is important for mammalian embryonic development. Although previous studies have reported that DNA methylation of a tandem repeated array sequence in paternal IG-DMR (IG-DMR-Rep) plays an essential role in the maintenance of DNA methylation in mice, the function of a tandem repeated array sequence in human IG-DMR (hRep) is unknown. Here, we generated mice with a human tandem repeated sequence, which replaced the mouse IG-DMR-Rep.

Nitrogen Availability and Microbial Communities of Canopy Soils in a Large Cercidiphyllum japonicum Tree of a Cool-Temperate Old Growth Forest.

Canopy soils on large trees are important for supporting the lives of many canopy plants, and thereby increasing regional biodiversity. However, because of the less accessibility to canopy soils, there is insufficient knowledge on how canopy soils produce available nitrogen (N) for canopy plants through the activity of canopy soil microbes. Canopy soils usually have different soil properties from ground soils, so we hypothesized that canopy soils would have unique microbial communities compared to ground soils, but still provide available N for canopy plants.

Identification of the first promoter-specific gain-of-function SOX9 missense variant (p.E50K) in a patient with 46,XX ovotesticular disorder of sex development.

SOX9, a transcription factor, is expressed in the undifferentiated XX and XY gonads. SRY induces significant upregulation of SOX9 expression in XY gonads. Loss-of-function SOX9 variants cause testicular dysgenesis in 46,XY patients, while duplication of the total gene or the upstream regulatory region results in testicular development in 46,XX patients.

A Quick Immuno-FISH Protocol for Detecting RNAs, Proteins, and Chromatin Modifications.

Immunofluorescence and fluorescence in situ hybridization (FISH) are widely used cytogenetic techniques for visualization of protein and RNA/DNA molecules. Here, we describe an experimental procedure for quick sequential immunofluorescence and RNA FISH (immuno-FISH), which enables the simultaneous detection of proteins, chromatin modifications, and RNAs on the inactive X-chromosome (Xi) using female mouse embryonic fibroblast (MEF) and tail-tip 3T3 cell lines. Using a pooled array of oligonucleotides labeled with a single fluorophore as an RNA FISH probe, we can reduce the time for RNA FISH from an overnight process to 1-2 h without losing its sensitivity.

The indirect γ-aminobutyric acid (GABA) receptor agonist gabaculine-induced loss of the righting reflex may inhibit the descending analgesic pathway.

In the spinal cord, γ-aminobutyric acid (GABA) interneurons play an essential role in antinociception. However, not all actions of GABA favor antinociception at the supraspinal level. We previously reported that gabaculine, which increases endogenous GABA in the synaptic clefts, induces loss of the righting reflex (LORR) that is one indicator of hypnosis, but not immobility in response to noxious stimulus.

The effect of induced mood on word imagery processing: An ERP study.

The effect of induced mood on language processing has been examined in behavioral and event-related potential (ERP) studies. A previous study examined the effects of induced mood on word imagery processing by the N400 and N700 components of the ERP and behavioral performance in an imageability judgment task in which participants decided whether a word easily evoked visual imagery or not (Ogawa and Nittono, 2019). The N400 amplitude was larger (more negative-going) under positive mood than under negative mood, while reaction time and the N700 amplitude were not affected by induced moods.

Structural Insights into the Substrate Specificity Switch Mechanism of the Type III Protein Export Apparatus.

Bacteria use a type III protein export apparatus for construction of the flagellum, which consists of the basal body, the hook, and the filament. FlhA forms a homo-nonamer through its C-terminal cytoplasmic domains (FlhA) and ensures the strict order of flagellar assembly. FlhA goes through dynamic domain motions during protein export, but it remains unknown how it occurs.

Mapping of a responsible region for sex reversal upstream of Sox9 by production of mice with serial deletion in a genomic locus.

Sox9 plays critical roles in testis formation. By mapping four familial cases of disorders of sexual development, a 32.5 kb sequence located far upstream of SOX9 was previously identified as being a commonly deleted region and named the XY sex reversal region (XYSR).

Single vs. repeated matrix metalloproteinase-13 knockdown with intra-articular short interfering RNA administration in a murine osteoarthritis model.

Our aims were 1) to estimate the duration of short interfering RNA (siRNA) effect on matrix metalloproteinase-13 () levels by a single intra-articular injection using a mouse knee osteoarthritis (OA) model and 2) to test whether repeated injections results in any additional suppressive effect on cartilage degradation compared to a single injection. OA was induced in 9 weeks old male C57BL/6 mice by destabilization of medial meniscus (DMM). Chemically modified siRNA targeted for was injected into the knee joint at 1 week post-DMM surgery.

Total Synthesis of (+)-Pyrenolide D.

An efficient approach to stereoselective construction of a spiro-γ-lactone core structure via BF-promoted formal [3 + 2] annulation of aldehydo-aldose derivatives with γ-methylene-γ-butyrolactone has been developed. The spiro-γ-lactone derivative was then used in an efficient total synthesis of (+)-pyrenolide D. The developed chemistry paves the way for total synthesis of structurally diverse natural products containing spiro-lactone cores.