Proximal Region Research Articles

Tectono-thermal evolution from the proximal to hyperextended domains of the northern South China Sea margin since initial rifting

The Pearl River Mouth Basin (PRMB) is located on the northern margin of the South China Sea (SCS). Heat flow measurements indicate that the PRMB is a typical ‘hot basin' characterized by a high background heat flow. However, the tectono-thermal evolution of the PRMB and the mechanisms involved are still controversial due to the different input parameters used in tectono-thermal models, small datasets and the limited area of the basin studied. We used tectono-thermal modelling with a multi-stage finite stretching model, constrained by extensive well data, to systematically analyse the tectono-thermal evolution of the PRMB since the onset of rifting. The study area was expanded relative to previous studies to cover both the proximal and hyperextended regions of the northern SCS margin, providing a more comprehensive understanding of its tectono-thermal history. Numerous wells and seismic data covering the entire basin were used, along with appropriate input parameters, to address the gaps in previous studies and to enhance the accuracy of the results. Our findings indicate that the PRMB underwent two phases of heating, primarily due to thinning caused by lithospheric extension during rifting. By the end of rifting, the Northern Depression Zone and the Kaiping Sag had reached peak heat flow, while the Panyu Lower Uplift and Baiyun Sag saw their highest heat flow since the initial rifting. The PRMB experienced thermal decay during the post-rift stage, but a significant reheating event occurred from 23.03 to 13.82 Ma, likely due to northwards lower crustal flow from the Baiyun Sag. The Panyu Lower Uplift and the Baiyun Sag had reached their peak heat flow by 13.82 Ma and the heat flow generally increased by 1–3 mW m −2 in the other studied areas during this stage. The basin's thermal decay slowed at 5.33 Ma and localized heating events linked to magmatic activities in the southern Dongsha Uplift were observed. In general, the proximal domain of the northern SCS margin reached peak heat flow by the end of rifting, whereas the hyperextended domain reached peak heat flow by 13.82 Ma. The heat flow in the hyperextended domain was generally higher than that in the proximal domain as a result of the thinner crust of the hyperextended domain caused by intense multiple lithospheric detachment–extensional thinning processes. During the rifting stage, the lithospheric extensional thinning process was the most important factor influencing the tectono-thermal evolution of the northern SCS margin, while lower crustal flow and magmatism were the most important factors during the post-rift stage.

Requirement of the N-terminal region of nonstructural protein 1 in cis for SARS-CoV-2 defective RNA replication.

SARS-CoV-2 belongs to the family Coronaviridae and carries a single-stranded positive-sense RNA genome. During coronavirus (CoV) replication, defective or defective interfering RNAs that lack a large portion of the genome often emerge. These defective RNAs typically carry the necessary RNA elements that are required for replication and packaging. We identified the minimum requirement of the 5' proximal region necessary for viral RNA replication by using artificially generated SARS-CoV-2 minigenomes. The minigenomes consist of the 5'-proximal region, an open reading frame (ORF) that encodes a fusion protein consisting of the N-terminal of viral NSP1 and a reporter gene, and the 3' untranslated region of the SARS-CoV-2 genome. We used a modified SARS-CoV-2 variant to support replication of the minigenomes. A minigenome carrying the 5' proximal 634 nucleotides replicated, whereas those carrying shorter than 634 nucleotides did not, demonstrating that the entire 265 nt-long 5' untranslated region and N-terminal portion of the NSP1 coding region are required for the minigenome replication. Minigenome RNAs carrying a specific amino acid substitution or frame shift insertions in the partial NSP1 coding sequence abrogated minigenome replication. Introduction of synonymous mutations in the minigenome RNAs also affected the replication efficiency of the minigenomes. These data suggest that the expression of the N-terminal portion of NSP1 and the primary sequence of the 5' proximal 634 nucleotides are important for minigenome replication.IMPORTANCESARS-CoV-2, the causative agent of COVID-19, is highly transmissible and continues to have a significant impact on public health and the global economy. While several vaccines mitigate the severe consequences of SARS-CoV-2 infection, mutant viruses with reduced reactivity to current vaccines continue to emerge and circulate. This study aimed to identify the minimal 5' proximal region of SARS-CoV-2 genomic RNA required for SARS-CoV-2 defective RNA replication and investigate the importance of an ORF encoded in these defective RNAs. Identifying cis-acting replication signals of SARS-CoV-2 genomic RNA is critical for the development of antivirals that target these signals. Additionally, replication-competent defective RNAs can serve as therapeutic reagents to interfere with SARS-CoV-2 replication. Our findings provide valuable insights into the mechanisms of SARS-CoV-2 RNA replication and the development of reagents that suppress SARS-CoV-2 replication.

Regional morphological adaptations of vastus lateralis muscle in response to different progressive resistance training programs: A randomised controlled trial.

Resistance training often increases muscle size, a phenomenon known as muscle hypertrophy. These morphological adaptations were typically documented to occur in a non-uniform pattern. Investigating the specific morphological adaptations to different training programs was of interest. This study aimed to investigate two resistance training programs, a high-intensity program (HI) and a combined high-intensity with low-intensity blood flow restriction program (MIX), on morphological adaptations of vastus lateralis muscle in healthy young men. Eighteen active participants were recruited and randomly assigned to the HI (n = 10) or MIX (n = 8) groups, undergoing different 6-week resistance training programs. The training volume set was equated and progressively increased from three sets in weeks 1 and 2 to six sets, and eight sets in weeks 3-4 and 5-6, respectively. Three specific regions of vastus lateralis were assessed by magnetic resonance imaging (MRI) and ultrasound imaging (US) during pre-and post-intervention. Statistical analysis revealed statistically significant increases in muscle area at the proximal (HI: Δ12%, MIX: Δ9.2%), middle (HI: Δ8.7%, MIX: Δ9.0%), and distal (HI: Δ14%, MIX: Δ13%) regions. Additionally, both HI and MIX groups showed statistically significant increases in the sum of muscle thickness post-intervention (HI: Δ12%, MIX: Δ19%) and in the sum of fascia thickness post-intervention (HI: Δ27%, MIX: Δ54%). Despite the MIX group training with higher volume load, no statistical differences were observed between groups for any week. These findings suggested that both HI and MIX programs effectively induced increases in muscle area and sums of muscle and fascia thickness in healthy young men, allowing practitioners to choose either program based on individual preferences and constraints.

Continuous periprosthetic bone loss around the TOP® cup and inferior survival rate at an 8-year follow-up: a prospective cohort study

BackgroundThe trabeculae-oriented pattern (TOP®) cup was designed to minimize acetabular periprosthetic bone loss. In our previous prospective study comprising 30 patients with a two-year follow-up we found a substantial decrease in periprosthetic bone mineral density (pBMD) in the proximal and medial regions of the TOP cup. The present study aims to investigate pBMD changes in the mid-term and how this affects implant survival.MethodsWe followed the previous cohort and estimated implant survival by Kaplan-Meier analysis, evaluated pBMD with dual-energy X-ray absorptiometry (DXA) and clinical outcome using the Harris Hip Score (HHS).ResultsMean follow-up was 8.6 (range 7.8–9.1) years. The eight-year implant survival rate for cup revision for all reasons was 83% (95% confidence interval {CI}: 70–97) and 86% (CI: 74–99) when cup revision due to aseptic loosening was the endpoint. Mean HHS at eight years was 95 (range 77–100). A further 12% (CI: 5–17) loss in pBMD was detected in the proximal Digas zone 1 and 12% (CI: 7–17) loss in Digas zone 2 also between two and eight years after surgery. pBMD continued to decrease up to 30% (CI: 24–36) in Digas zones 1, 2 and 3 compared to pBMD immediately postoperatively.ConclusionsThe TOP cup shows inferior mid-term survival rates compared to other uncemented cups, as well as a continuous decrease in pBMD. Periprosthetic bone loss cannot be prevented by this uncemented cup.Clinical trial numberNot applicable.

Sexually dimorphic renal expression of mouse Klotho is directed by a kidney-specific distal enhancer responsive to HNF1b

Transcription enhancers are genomic sequences regulating common and tissue-specific genes and their disruption can contribute to human disease development and progression. Klotho, a sexually dimorphic gene specifically expressed in kidney, is well-linked to kidney dysfunction and its deletion from the mouse genome leads to premature aging and death. However, the sexually dimorphic regulation of Klotho is not understood. Here, we characterize two candidate Klotho enhancers using H3K27ac epigenetic marks and transcription factor binding and investigate their functions, individually and combined, through CRISPR-Cas9 genome engineering. We discovered that only the distal (E1), but not the proximal (E2) candidate region constitutes a functional enhancer, with the double deletion not causing Klotho expression to further decrease. E1 activity is dependent on HNF1b transcription factor binding site within the enhancer. Further, E1 controls the sexual dimorphism of Klotho as evidenced by qPCR and RNA-seq. Despite the sharp reduction of Klotho mRNA, unlike germline Klotho knockouts, mutant mice present normal phenotype, including weight, lifespan, and serum biochemistry. Lastly, only males lacking E1 display more prominent acute, but not chronic kidney injury responses, indicating a remarkable range of potential adaptation to isolated Klotho loss, especially in female E1 knockouts, retaining renoprotection despite over 80% Klotho reduction.

The transcription factors NR5A1 and JUNB cooperate to activate the Axl promoter in mouse Sertoli cell lines.

The Axl gene is a receptor tyrosine kinase essential for male fertility. With other Tyro3 family members, it regulates cell apoptosis and preserves the organization of seminiferous tubules. However, the regulation of the expression of Axl in testicular Sertoli cells is not entirely understood. The transcription factors NR5A1 and JUNB are involved in several male fertility mechanisms such as sex development and steroidogenesis. We hypothesize that Axl promoter activity is regulated by cooperation between JUNB and NR5A1 in Sertoli cells. Following transfections of TM4 Sertoli cells with DsiRNA interference against Axl, our results show that cell morphology may be regulated by AXL. Using transfections of expression plasmids and reporter plasmids containing the Axl promoter, we report that Axl expression is highly activated by cooperation between NR5A1 and JUNB in TM4 and 15P-1 Sertoli cells. Chromatin immunoprecipitation and luciferase reporter assays with 5' promoter deletions demonstrate that JUNB and NR5A1 are being recruited to DNA regulatory elements in the proximal region of the Axl promoter. The fourth intronic region of Axl also participates in the recruitment of JUNB. Thus, Axl expression is regulated by a cooperation between the transcription factors JUNB and NR5A1 and influences the morphology of TM4 Sertoli cells.

The Structure of Cilium Inner Junctions Revealed by Electron Cryo-tomography.

The cilium is a microtubule-based organelle critical for many cellular functions. Its assembly initiates at a basal body and continues as an axoneme that projects out of the cell to form a functional cilium. This assembly process is tightly regulated. However, our knowledge of the molecular architecture and the mechanism of assembly is limited. By applying electron cryotomography and subtomogram averaging, we obtained subnanometer resolution structures of the inner junction in three distinct regions of the cilium: the proximal region of the basal body, the central core of the basal body, and the flagellar axoneme. The structures allowed us to identify several basal body and axoneme components. While a few proteins are distributed throughout the entire length of the organelle, many are restricted to particular regions of the cilium, forming intricate local interaction networks and bolstering local structural stability. Finally, by knocking out a critical basal body inner junction component Poc1, we found the triplet MT was destabilized, resulting in a defective structure. Surprisingly, several axoneme-specific components were found to "infiltrate" into the mutant basal body. Our findings provide molecular insight into cilium assembly at its inner Junctions, underscoring its precise spatial regulation.

Telomere-to-telomere, gap-free assembly of the Rosa rugosa reference genome

Rosa, a genus esteemed worldwide for its ornamental plants, has encountered barriers in functional genomic studies and further genetic enhancement due to incomplete sequences and floating regions in previously sequenced genomes. Our groundbreaking study introduced a meticulously assembled, continuous, and fully bridged reference genome for Rosa rugosa, constructed through a sophisticated combination of PacBio High-Fidelity, ONT ultra-long reads, and Hi-C data. This robust assembly spanned 444.55 Mb and encompassed 34 109 protein-coding genes. We have uniquely assembled each chromosome into single, gap-free structures, successfully identifying all 14 telomeres and seven centromeres, a feat not achieved previously. The centromeric regions were distinguished by tandem repeats, primarily composed of centromere-specific 159-bp monomers, and a significant enrichment of ATHILA/Gypsy long terminal repeat retrotransposons in proximal regions. Our research highlighted recent tandem duplications as instrumental in bolstering R. rugosa's stress tolerance, environmental adaptability, and enhanced anthocyanin synthesis. Furthermore, our study ventured into uncharted territory by predicting transcription factors potentially regulating anthocyanin biosynthesis through the employment of gene co-expression networks, providing new avenues for research. This comprehensive reference genome not only serves as a cornerstone for in-depth exploration of genomic architecture and functionalities in R. rugosa but also acts as a catalyst for innovative breeding strategies and genetic refinement within the genus.

Robust optimization of geometrical properties of flow diverter stents for treating cerebral aneurysm: A proof-of-concept study

This study presents a novel approach to optimize the design of flow diverter (FD) stents for cerebral aneurysm (CA) treatment. By addressing sources of uncertainty in cardiovascular simulations, including geometrical and physical properties and boundary conditions, we aim to assess the applicability of robust optimization techniques to the FD design, establishing a foundation for acquiring robust FDs that are capable of operating consistently under various real-world scenarios. Blood flow in a simplified 2-dimensional CA and FD model was simulated through computational fluid dynamics (CFD). A design space exploration method, incorporating Latin hypercube sampling and Kriging surrogate models, was employed to obtain the optimal solution. The objective was to maximize the reduction in velocity and vorticity within the CA sac. This study used non-intrusive polynomial chaos expansion (PCE) to quantify and propagate the input uncertainties through the computational model and compute the statistical moments of velocity and vorticity reductions. To assess the effect of uncertain sources on objective functions, a sensitivity analysis method based on Sobol indices was applied. Robust optimization involved simultaneously optimizing the mean and standard deviation of velocity reduction. Additionally, we accounted for patients’ specific conditions and repeated the robust optimization. The results indicate that blood Hematocrit and inlet velocity are the most impactful uncertain sources in FD optimization. Moreover, the obtained Pareto front shows that in robust designs, FD struts are concentrated in the distal region of the CA neck, while optimal designs have more struts in the proximal region. This study proposes an FD that compromises robustness and optimality with a velocity reduction of 72.31 % and a standard deviation of 0.00343.

L3MBTL2 maintains MYCN-amplified neuroblastoma cell proliferation through silencing NRIP3 and BRME1 genes.

Epigenetic alterations critically affect tumor development. Polycomb-group complexes constitute an evolutionarily conserved epigenetic machinery that regulates stem cell fate and development. They are implicated in tumorigenesis, primarily via histone modification. Polycomb repressive complex 1 (PRC1) complexes 1-6 (PRC1.1-6) mediate the ubiquitination of histone H2A on lysine 119 (H2AK119ub). Here, we studied the functional roles of a PRC1.6 molecule, L3MBTL2, in neuroblastoma (NB) cells. L3MBTL2-knockout and knockdown revealed that L3MBTL2 depletion suppressed NB cell proliferation via cell-cycle arrest and gamma-H2A.X upregulation. L3MBTL2-knockout profoundly suppressed xenograft tumor formation. Transcriptome analysis revealed suppressed cell-cycle-related and activated differentiation-related pathways. Break repair meiotic recombinase recruitment factor 1 (BRME1) and nuclear receptor interacting protein 3 (NRIP3) were notably de-repressed by L3MBTL2-knockout. The deletion of L3MBTL2 reduced enrichment of H2AK119ub and PCGF6 at transcriptional start site proximal regions of the targets. Add-back studies unveiled the importance of L3MBTL2-BRME1 and -NRIP3 axes for NB cell proliferation. We further manifested the association of MYCN with de-repression of NRIP3 in an L3MBTL2-deficient context. Therefore, this study first revealed the significance of L3MBTL2-mediated gene silencing in MYCN-amplified NB cells.

Digital Papillary Adenocarcinoma: Uncommon Malignancy of Sweat Glands - Two Rare Cases

Abstract Digital papillary adenocarcinoma (DPA) is a very rare malignant tumor of the sweat glands originally described by Helwig in 1979.It often presents as a solitary painless mass, commonly found on the volar surface of the fingers and toes and the adjacent skin of the palms and soles. Materials and Methods: Two rare cases of DPA in 43- and 48-year-old male patients who presented with swelling finger were included over period of one year study in a tertiary care hospital. Results: Our study had two cases with male patient of age 48 year and 43 year old. Both presented with swelling and a mass lesion left middle finger, past 8 years and nodule in proximal phalangeal region of left index finger for 15 years respectively. On histomorphology and immunohistochemistry both were diagnosed as DPA. Conclusion: DPA is a rare adnexal tumor which generally present for months to years before the patient seeks medical attention. Careful evaluation with early surgical removal and accurate histological diagnosis is must when managing patients with DPA as these can be aggressive and even result in metastasis..

Short photoperiod inhibited gonadal growth and elevated hypothalamic Dio3 expression unrelated to promoter DNA methylation in young Brandt's voles.

Photoperiod, the length of daylight, has a significant impact on the physiological characteristics of seasonal breeding animals, including their somatic and gonadal development. In rodents, expression of deiodinase type II (Dio2) and III (Dio3) in the hypothalamus is crucial for responding to photoperiodic signals. However, research on the photoperiodism of hypothalamic gene expression and the corresponding regulatory mechanism in Brandt's voles living in the Mongolian steppes is limited. In this study, we gradually changed day length patterns to simulate spring (increasing long photoperiod, ILP) and autumn (decreasing short photoperiod, DSP). We compared the somatic and gonadal development of voles born under ILP and DSP and the expression patterns of five reproduction-related genes in the hypothalamus of young voles. The results showed that DSP significantly inhibited somatic and gonadal development in both female and male offspring. Compared with ILP, Dio3 expression was significantly upregulated in the hypothalamus under DSP conditions and remained elevated until postnatal week 8 in both males and females. However, there was no significant difference in the methylation levels of the proximal promoter region of Dio3 between ILP and DSP, suggesting that methylation in the proximal promoter region may not be involved in regulating the expression of Dio3. These findings suggest that hypothalamic expression of Dio3 plays a key role in the photoperiodic regulation of gonadal activity in Brandt's voles. However, it appears that CpGs methylation in the promoter region is not the main mechanism regulating Dio3 expression.

Application of 3D-printed individualized porous tantalum buttress in shelf acetabuloplasty for developmental dysplasia of the hip

Shelf acetabuloplasty is a viable treatment for developmental dysplasia of the hip (DDH), yet autologous bone grafts have faced challenges such as morphological mismatch, imprecise placement, and long-term graft resorption. The rapid advancement of three-dimensional (3D) printing technology in orthopedics has enabled the acquisition of digital image correlation method (DICM) data from patient pelvic to bilateral femoral proximal regions through computed tomography (CT) scans. This data is then utilized by software to create 3D models and design individualized metal implants. Using tantalum-based TA1 type I spherical powder as the raw material, a porous tantalum buttress with a pore size of 600 microns and a porosity of 75% was fabricated employing the selective electron beam melting (SEBM) technology. Our Center has applied this 3D-printed individualized porous tantalum buttress in shelf acetabuloplasty for 21 DDH patients (25 hips), including 6 males and 15 females, with a mean age of (21.38±7.39) years. The follow-up period ranged from 12 to 47 months, averaging (22.64±10.86) months. At the final follow-up, patient-reported outcomes (PROs) were used to assess patient’s subjective feelings. Significant improvements were observed in the Non-Arthritic Hip Score (NAHS), modified Harris Hip Score (mHHS), Hip Outcome Score-Sports Subscale (HOS-SSS), International Hip Outcome Tool-12 (iHOT-12), and Visual Analog Scale (VAS) compared with preoperative levels (P<0.01). Radiographic assessments showed that postoperative lateral center edge (LCE) angle, Tonnis angle, acetabular angle (Sharp’s angle), and femoral head coverage all trended towards normal, with statistically significant differences (P<0.01). There was no implant displacement or bone resorption in any case, and no progression in Tonnis grading for hip osteoarthritis (OA) was noted postoperatively, with an 84% patient satisfaction rate. The early follow-up results of 3D-printed individualized porous tantalum buttress in shelf acetabuloplasty are satisfactory, indicating a reliable application prospect for the treatment of DDH patients.

Human‐initiated autocyclic delta failures

ABSTRACTRiver regulations have resulted in changes in the hydrology and particle budgets of fluvial systems. Since the 19th century, many rivers have been significantly modified to control flood hazards, to gain land from swamp areas for agricultural purposes, and to stabilize river‐levels and lake‐levels to facilitate navigation. These dramatic changes of the river courses have impacted the sediment budgets and grain‐size dissemination along them as well as the sediment distribution at the delta mouths in the downstream lakes, which could lead to slope instabilities. Deposits of such catastrophic lacustrine mass movements caused by delta collapses have been, for instance, observed in Lake Brienz (Switzerland), where relatively thick (0.5 to 1.3 m) and voluminous (>1 million m3) megaturbidites are stacked in the deep basin witnessing these processes. This study uses sediment cores and seismic data to reconstruct the megaturbidites' history in Lake Brienz. Data reveal that mass‐movement deposits, originating from the Aare Delta, one of the two main inflows, have mean ages of 1853, 1905, 1942 and 1996 ce and that they were unprecedented in, at least, half a millennium. The fact that the numbers of floods and earthquakes have not changed radically over this time period implies that human impact is the most likely explanation for these failure events. Therefore, the recurrent delta collapses are attributed to the focused sediment accumulation at the front of the channelized inflow in the proximal delta region, caused by the modification of the Aare River through its straightening and channelization during the late 19th century. These findings indicate that river regulation can affect delta sedimentation, leading to autocyclic delta collapses. Those collapses, in turn, can potentially generate tsunami waves, representing an additional natural hazard for shoreline communities.

Use of VAC System and Dermal Matrix as a Surgical Alternative for Lower Extremity Degloving Treatment

Introduction: Soft tissue degloving injuries are the result of high-energy forces that produce compression, twisting or friction of tissues causing soft tissue avulsion, in which a large portion of skin and subcutaneous tissue is detached from the underlying fascia and muscles with musculocutaneous damage and perforating fasciocutaneous vessels. Case Report: The case of a 67-year-old female patient was reported, who as a result of a traffic accident (run over) suffered cutaneous loss (degloving) of the left lower limb. The patient underwent surgical cleaning with debridement of dead tissue plus full-thickness skin autograft for partial coverage of the exposed area in the proximal region. At the same time the surgery was performed, the VAC system was placed. After multiple surgical cleanings, scarce granulation tissue was revealed in the middle and distal third of the left leg. Therefore, it was decided to cover the exposed area with dermal matrix for later definitive coverage. The patient is being followed up by outpatient service. It is revealed that the patient already performs assisted mobility, and the grafts are assimilated. Conclusions: Traffic accidents are a major problem worldwide, especially those of high-energy may require multidisciplinary treatment, prolonged surgical time until recovery and return to daily activities. The initial treatment for degloving has to be performed by trained professionals since the criterion for assessing flap vitality is indispensable. When complex skin grafts are not an option, advanced wound treatment should be used, as in this case, where VAC therapy and permanent synthetic substitute were used for the total coverage of a major defect with lost dermal tissue.

Progesterone may be a regulator and B12 could be an indicator of the proximal D4Z4 repeat methylation status on 4q35ter.

Facioscapulohumeral dystrophy (FSHD) has a hypomethylation-related epigenetic background and exhibits a different course in male and female patients. The differences between males and females have been linked to the levels of sex hormones. This study is the first to investigate the possible effect of these hormones on methylation status. We hypothesized that the levels of sex-related hormones, estradiol, testosterone, progesterone, and prolactin might be associated with the methylation status of the proximal part of the D4Z4. We also investigated the effect of fT3, folic acid, and vitamin B12 levels. We collected blood from 28 FSHD patients and 28 controls. DNA was extracted from each individual for bisulfite methylation analysis and serum was separated for biochemical analysis of estradiol, testosterone, progesterone, prolactin, fT3, folic acid, and B12 analysis. Methylation analysis was specified to the DR1, 5P regions and the proximal region covering both DR1 and 5P. Methylation levels were compared between FSHD patients and controls. The correlation of methylation levels with estradiol, testosterone, progesterone, prolactin, fT3, folic acid, and B12 was investigated. We found that the 5P region and the proximal region were significantly hypomethylated in FSHD patients compared to the controls, but not the DR1 region. Male patients exhibited a significant reduction in DNA methylation compared to male controls. Older FSHD patients exhibited a notable decrease in fT3 levels and hypomethylation of the 5P region. Analyses of each CpG revealed seven hypomethylated positions that were significantly different from the control group. Two of the positions demonstrated a correlation with progesterone in the control group. With the exception of one position, the methylation levels were inversely correlated with vitamin B12 in FSHD patients. The results of our study indicate that the methylation of the proximal D4Z4 region, particularly at specific positions, may be associated with progesterone. In addition, vitamin B12 may be an indicator of hypomethylation. We suggest that examining position-specific methylations may be a useful approach for the development of epigenetic treatment modalities.

No Difference in Graft Signal Intensity on Magnetic Resonance Imaging or Clinical Outcome Between Anterior Cruciate Ligament Reconstruction With and Without Suture Augmentation

PurposeTo evaluate the safety of anterior cruciate ligament reconstruction (ACLR) with suture augmentation (SA) through clinical evaluations, monitoring of complications, and evaluation of early graft remodeling using magnetic resonance imaging (MRI). MethodsData of participants who underwent anatomic double-bundle ACLR using hamstring tendon autografts, with a minimum 2-year follow-up, were retrospectively reviewed. Participants undergoing ACLR with SA were propensity-matched with those without SA. Postoperative clinical evaluations, including Lysholm and Tegner activity scores, International Knee Documentation Committee (IKDC) evaluation scores, KT-1000 side-to-side difference for knee stability at a 2-year follow-up, and complications, were recorded. The minimal clinically important difference (MCID) was calculated for IKDC scores. The anteromedial bundle graft was categorized into distal, middle, and proximal regions on the 6-month and 1-year postoperative MRI. The signal-to-noise quotient (SNQ) of each region of interest was calculated to evaluate the graft signal intensity, which was compared between the two groups using Welch’s t-test. ResultsFifty-three participants were included in each group. No significant differences were observed between the non-SA and SA groups in the Lysholm scores (96.6 ± 6.2; 95.3 ± 5.8, P = 0.25), Tegner activity scores (4.8 ± 1.4; 5.0 ± 1.3, P = 0.49), IKDC scores (90.4 ± 10.8; 87.1 ± 12.9, P = 0.15), percentage of patients meeting the MCID (94.3%; 83.0%, P = 0.12), or postoperative KT-1000 side-to-side difference (0.9 ±1.2; 0.7 ± 1.8 mm, P = 0.56). One (1.8%) and two (3.7%) cases of re-tears occurred in the non-SA and SA groups, respectively; no other complications occurred. The postoperative SNQ measurements revealed no significant differences in signal changes in all regions of the grafts between the two groups. ConclusionsThis study confirmed no difference in graft signal intensity on MRI or clinical outcome between ACLR with and without SA at the 2-year follow-up. Level of EvidenceLevel III, retrospective, comparative study.

CREB activates the MafA promoter through proximal E-boxes and a CCAAT motif in pancreatic β-cells.

MafA is a key transcriptional regulator of pancreatic islet β-cell function. Its target genes include those encoding preproinsulin and the glucose transporter Glut2 (Slc2a2); thus, MafA function is essential for glucose-stimulated insulin secretion. Expression levels of MafA are reduced in β-cells of diabetic mouse models and human subjects, suggesting that β-cell dysfunction associated with type 2 diabetes is attributable to the loss of MafA. On the other hand, MafA is transcriptionally upregulated by incretin hormones through activation of CREB and its co-activator CRTC2. β-cell-specific expression of MafA relies on a distal enhancer element. However, the precise mechanism by which CREB-CRTC2 regulates the enhancer and proximal promoter regions of MafA remains unclear. In this report, we analyzed previously published ChIP-seq data and found that CREB and NeuroD1, a β-cell-enriched transactivator, bound to both the promoter and enhancer regions of human MAFA. A series of reporter assays revealed that CREB activated the enhancer through a conserved cAMP-responsive element (CRE) but stimulated MAFA promoter activity even when the putative CRE was deleted. Two E-box elements and a CCAAT motif, which bind NeuroD1 and ubiquitous NF-Y transcription factors, respectively, were necessary for transcriptional activation of the MAFA promoter by CREB. Genome-wide analysis of CREB-bound loci in β-cells revealed that they were enriched with CCAAT motifs. Furthermore, promoter analysis of the Isl1 gene encoding a β-cell-enriched transcription factor revealed that a CRE-like element and two CCAAT motifs, but not the E-box, were necessary for activation by CREB. These results provide clues to elucidate the detailed mechanism by which CREB regulates MafA as well as β-cell-specific genes.

Strontium Isotopic Composition as Tracers for Identifying Groundwater Recharge Sources in the Choushui River Alluvial Plain, Western Taiwan

Groundwater is a vital resource in the Chuoshui River alluvial plain (CSAP), a key agricultural area in Taiwan. Understanding groundwater recharge is crucial for sustainable water management amidst changing climatic conditions and increasing water demand. This study investigates the major ion composition, solute Sr concentrations, and 87Sr/86Sr ratios in groundwater and stream water from the Choushui River (CSR) to trace groundwater recharge sources. The Piper diagram reveals that most groundwater samples are of the freshwater Ca–HCO3 type, aligning with the total dissolved solids (TDS) classification. TDS and major ion compositions indicate that groundwater near Baguashan Terrace (BGT) and Douliu Hill (DLH) primarily derives from stream water and rainwater. Na+ and Cl− enrichment in some aquifers of BGT and DLH is attributed to the dissolution of paleo-sea salt and mixing with paleo-seawater from sedimentary porewater. Elevated dissolved Sr concentrations and lower 87Sr/86Sr ratios in these aquifers further support the intrusion of paleo-seawater. Groundwater in the proximal fan shows high TDS due to intensive weathering, complicating the use of TDS as a tracer. Sr isotopic compositions and solute Sr2+ concentrations effectively distinguish recharge sources, revealing that the CSR mainstream primarily recharges the proximal fan and BGT region, while CSR tributaries and rainwater mainly recharge the DLH region. This study concludes that Sr isotopic compositions and solute Sr2+ concentrations are more reliable than TDS and major ion compositions in identifying groundwater recharge sources, enhancing our understanding of groundwater origins and the processes affecting water quality.

The dual role of a highly structured RNA (the S fragment) in the replication of foot-and-mouth disease virus.

Secondary and tertiary RNA structures play key roles in genome replication of single-stranded positive sense RNA viruses. Complex, functional structures are particularly abundant in the untranslated regions of picornaviruses, where they are involved in initiation of translation, priming of new strand synthesis and genome circularization. The 5' UTR of foot-and-mouth disease virus (FMDV) is predicted to include a c. 360 nucleotide-long stem-loop, termed the short (S) fragment. This structure is highly conserved and essential for viral replication, but the precise function(s) are unclear. Here, we used selective 2' hydroxyl acetylation analyzed by primer extension (SHAPE) to experimentally determine aspects of the structure, alongside comparative genomic analyses to confirm structure conservation from a wide range of field isolates. To examine its role in virus replication in cell culture, we introduced a series of deletions to the distal and proximal regions of the stem-loop. These truncations affected genome replication in a size-dependent and, in some cases, host cell-dependent manner. Furthermore, during the passage of viruses incorporating the largest tolerated deletion from the proximal region of the S fragment stem-loop, an additional mutation was selected in the viral RNA-dependent RNA polymerase, 3Dpol. These data suggest that the S fragment and 3Dpol interact in the formation of the FMDV replication complex.