bluuebunny/crossref_metadata_2025_split huggingface dataset:
Accelerometer Assessment of Spinal Joint Crepitus Before and After Spinal Manipulation
Authors: Cramer Gregory D , Budavich Matthew , Bora Preetam , Ross Kim 
Published: November 2023

The objective of this IRB‐approved project was to assess spinal joint [facet joint, zygapophyseal (Z) joint] vibrations/sounds (crepitus) during lumbar motion before and after side‐posture spinal manipulation (SM).MethodsUsing methods previously developed to assess joint cavitation, 9 accelerometers were applied to the lumbar region of 5 healthy and 5 low back pain (LBP) subjects, allowing assessment of crepitus. Subjects underwent full lumbar ranges of motion, SM, and repeated ranges of motion, while accelerometer recordings were made. Subjects were also divided into 3 age groups for comparisons (18–25, 26–45, and 46–65 years). Primary outcomes included: incidence of crepitus, crepitus prevalence (average number of crepitus events/subject), crepitus amplitude, and motion(s) producing the most crepitus.ResultsThe accelerometry methods were successfully implemented on all subjects. Crepitus was identified as simultaneous deviations from baseline of multiple accelerometers. As in previous work, artifacts had a distinct appearance and were easily distinguished from crepitus. Overall, 23 instances of crepitus were recorded from 15 different Z joints. Crepitus prevalence decreased pre‐ to post‐SM (average pre= 1.4 crepitus/subject, range 0–7 crepitus/subject vs. post= 0.9, range 0–2). Crepitus prevalence progressively increased from the youngest to oldest age groups (pre‐SM= 0.0, 1.67, and 2.0, respectively, range 0–10; and post‐SM= 0.5, 0.83, and 1.5, range 1–5). Prevalence was higher in LBP subjects compared to healthy (pre‐SM‐LBP= 2.0, range 0–7, vs. pre‐SM‐healthy= 0.8, range 0–4; post‐SM‐LBP= 1.0, range 0–2 vs. post‐SM‐healthy= 0.8, range 0–2), even though healthy subjects were older than LBP (40.8 years vs. 27.8 years). Crepitus amplitude was <0.16 mV. Average pre‐SM crepitus amplitude was 0.08 mV, which was greater than post‐SM crepitus, which was 0.03 mV. The pre‐ vs. post‐SM crepitus difference was greater in LBP (pre‐SM 0.09 mV, post‐SM 0.04) than healthy subjects (pre‐SM 0.04 mV, post‐SM 0.03 mV). Flexion produced the most crepitus (pre‐SM eight, 57.1%, post‐SM six, 66.7%, and overall fourteen, 60.9%). For pre‐SM, flexion was followed by right lateral flexion (three, 21.4%) with extension, left lateral flexion and left rotation all producing the same amount of crepitus (one each, 7.1%). Post‐SM flexion was followed by extension (two, 22.2%) and right rotation (one, 11.1%), with no crepitus recorded in the other motions.ConclusionsCrepitus was greatest in flexion; prevalence increased with age, was higher in LBP than healthy subjects, and overall decreased following SM; crepitus amplitude was lower post‐SM than pre‐SM.Support or Funding InformationNIH/NCCIH Grant# – 3R01AT000123‐06S2
Metaphysis repair process differ from the diaphysis
Authors: Inoue Satoshi 
Published: November 2023

PurposeIn humans, fractures of the long bones mainly occur in the metaphysis such as distal radius. However, most studies of the bone repair process have focused on the diaphysis region, thus little has been reported on the metaphysis repair process. In this study, we compared the bone repair process between the metaphysis and the diaphysis of the mouse tibia.Materials and MethodsBone apertures were formed in the tibial metaphysis and diaphysis of mice by 0.8mm drill hole. At indicated times after the surgery, samples were collected, and examined by histomorphometrical (micro‐computed tomography), histological (toluidine blue), histochemical (alkaline phosphatase (ALP)), and immunohistochemical (osteocalcin and type I collagen) analyses. Quantitative real‐time PCR was used to analyze the expression of mRNAs in the periosteum (sox9, type 2 collagen) and bone marrow (osterix, runx2, type 1 collagen).ResultsIn the metaphysis, cartilage formation was not detected during the repair period on the periosteum side. The bone aperture was filled with newly formed bone developed from bone marrow at 7 days, and then gradually decreased. In the diaphysis, cartilage was formed around the bone aperture at day 4 and sequentially replaced by bone on the periosteal side. Periosteal callus peaked at day 14, and then remodeled to original architecture. Medullary callus gradually increased to day 14, and the aperture filled with medullary callus transiently at day 14, then medullary callus rapidly resorbed at day 21. In both sites, bone mineral density (BMD) was gradually restored at the bone aperture. At day 14, BMD was higher in the metaphysis than in the diaphysis, but the diaphysis showed significantly high BMD at day 42. In diaphysis, the expression of type 2 collagen mRNA reached to the peak at day 5 following the peak expression of sox9 mRNA reached to the peak at day 4 on the periosteum. However, in the metaphysis, expression of those mRNAs was lower than the diaphysis. Expression of the osteogenic markers in bone marrow, alkaline phosphatase, osteocalcin and type I collagen, appeared earlier in the metaphysis than the diaphysis. In the metaphysis, osterix, runx2 and type 1 collagen mRNAs were markedly upregulated compared with the diaphysis.DiscussionThese results indicate a difference in the bone repair process between the metaphysis and the diaphysis, suggesting a functional heterogeneity of the periosteum and bone marrow mesenchymal cells in response to bone fracture.
The Effects of Bone Morphogenetic Protein‐7 and Interleukin‐1 on Biomechanical Properties of Meniscus Explants
Authors: Taylor Victor Wayne , Hicks Justin , Ferguson Cristin , Danelson Kerry 
Published: November 2023

IntroductionThe meniscus is a wedge, C‐shape fibrocartilaginous tissue that is essential for the normal biomechanical function of the knee. Damage to the meniscus leads to degenerative changes and loss of function mechanical function of the knee. Cytokines like interleukin‐1 (IL‐1) can inhibit repair process. However, anabolic factors like bone morphogenetic protein‐7 (BMP‐7) may help aid in the repair process. The goal of this project is to observe the effects of BMP‐7 and IL‐1 on the biomechanical properties of the meniscus using microindentation test.MethodSix‐millimeter porcine meniscus explants were cut using a biopsy punch and cultured in 1% fetal bovine serum media for seven days. Explants were cultured without treatment (control group) or treated with BMP‐7, IL‐1 alpha, and or combined with BMP‐7 and IL‐1 alpha. Microindentation test was used to test the elasticity of the explant tissues.ResultsThe elasticity for the explants treated with BMP‐7 (2.480 MPa) were significant compared to explants treated with either IL‐1 alpha (1.501 MPa; p= .001) or both BMP‐7 and IL‐1 alpha (.866; p ≤ .000). This indicates BMP‐7 and IL‐1 have a significant effect on the meniscus biomechanical properties. BMP‐7 increase the elasticity (but not significant) compared to the control explants (2.072 MPa; p= .065), while IL‐1significantly decrease the elasticity (p= .017).ConclusionThis data shows that BMP‐7 has the potential to exert an anabolic effect on cultured meniscus explants reflected by the presence of increased mechanical compression properties. As a result, the meniscus explant tissues that were cultured with BMP‐7 became more elastic after being cultured for seven days. In addition, meniscus cultured with IL‐1 alpha or a combination of BMP‐7 and IL‐1 alpha became less elastic. Future experiments will continue testing the effect of these two treatments.
Comparative Anatomy of the Anterior Cruciate Ligament Insertions in Primates
Authors: Ruth Aidan , Raghanti Mary Ann , Lovejoy C. Owen 
Published: November 2023

Despite a robust body of clinical literature on human entheses, relatively little is known about their anatomy in other species. We sought to document the microstructure of the femoral and tibial insertions of the anterior cruciate ligament (ACL) in a number of mammals, with a particular focus on Primates. Species investigated included those using unguligrade (deer, sheep), digitigrade (lion, tiger), and plantigrade (gorilla, Diana monkey, human) postures. We also explored the relationships between fibrocartilage thickness at ligament insertion sites and body mass.We found that most anatomical landmarks of the human ACL insertion have homologues in other mammals, including the Lateral Intercondylar Ridge (LIR), the Lateral Intercondylar Posterior Ridge (LIPR), and the Medial Intercondylar Ridge of the Tibia. However, we found that these ridges are not limited termini as is often assumed. Rather, their entheses are expansive and exhibit fibers that extend from both anterior and posterior aspects of these ridges. Considerable variation exists at the lateral border of the tibial ACL insertion, most likely related to spatial distinction of three subdivisions of the ACL (“bundles”) in deer and other ungulates. While primates still display up to three ACL bundles, they adhere more closely to one another and lack distinct septa. Humans share this primate‐typical anatomy.We measured fibrocartilage thickness of the ACL enthesis on histological thin sections using the method of orthogonal intercepts. Results of a standard linear regression of fibrocartilage thickness and body size indicate that there is no relationship between the natural log of body size as reported in the literature and the natural log of fibrocartilage thickness for either the femoral (F = 1.505, p= 0.1) or tibial (F = 2.57, p = 0.09) insertion sites of the ACL. These data suggest the possibility that, despite the potential biomechanical advantage of substantial fibrocartilage thickness, there may be an upper limit dictated by the (in)ability of nutrients and other cellular signals to diffuse within its extracellular matrix.
New microRNA Biotechnology to Inhibit Inflammation and Regenerate Bone
Authors: Amendt Brad , Cao Huojun , Yu Wenjie , Holton Nathan , Eliason Steven , Hong Liu 
Published: November 2023

Current tools for the inhibition of microRNA (miR) function are limited to modified antisense oligonucleotides, sponges, and decoy RNA molecules and none have been used to understand miR function during development. CRISPR/Cas mediated deletion of miR sequences within the genome requires multiple chromosomal deletions to remove all functional miR families due to the duplication of miR seed sequences and family members. We report a novel plasmid‐based miR inhibitor system (PMIS) that inhibits miR family members in cells and mice. The PMIS engineered optimal secondary structure, flanking sequences and specific antisense miR oligonucleotide sequence bind the miR in a stable complex to inhibit miR activity. In cells, one PMIS can effectively inhibit miR family members that share the same seed sequence. A complete family of miRs can be inhibited with a single plasmid. Different PMIS miR inhibitors can be linked together to knockdown multiple miRs expressed from different chromosomes. The PMIS shows no off‐target effects or toxicity and is highly specific for miRs sharing identical seed sequences. PMIS constructs associate with Dicer and Argonaute to form a stable miR inhibitor complex. Transgenic mice expressing PMIS‐miRs reveal different developmental processes affected by miRs. Genome‐wide analyses of PMIS transgenic mice and cells identified new miR regulated gene networks. Complete inhibition of the miR‐17‐92, miR‐106a‐363, miR‐106b‐25, and miR‐200 clusters reveals new mechanisms for bone regeneration and developmental defects for these miRs. These miRs also control inflammation through the direct targeting of pro‐inflammatory cytokines. We report a new tool to dissect the role of miRs in development without genome editing and as a potential new therapeutic reagent.Support or Funding InformationThis work was supported by grants from the State of Iowa Bio Venture group, College of Dentistry and Carver College of Medicine and NIH DE025328 to LH.
Direct Conversion of Oral Epithelial Cells to Progenitor Dental Mesenchymal Cells by a Novel microRNA‐Transcription Factor Pathway
Authors: Mansaray Maurisa N , Sharp Thad , Li Xiao , Huojun Cao , Amendt Brad 
Published: November 2023

There are many protocols used for regeneration therapies to regenerate tissues and structures. To date, tooth regeneration practices rely heavily on the use of stem cells, however, these can be difficult to obtain and culture. We have previously introduced a new method of cellular reprogramming by over expression of the transcription factor, Pitx2 and microRNA‐200a‐3p. We now demonstrate that over expression of the Paired Box 9 (Pax9) gene and inhibition of miR‐200a using the Plasmid‐based miRNA Inhibitor System (PMIS‐miR‐200a), converts oral epithelial cells to progenitor dental mesenchyme cells. Transgenic DNA was introduced using the lentivirus packaging system. The microRNA inhibitor system PMIS‐miR‐200a was constructed by NaturemiRI. A two‐staged approach is used to generate stable oral epithelial cells expressing PAX9, cells were sorted and after several weeks were transduced with PMIS‐miR‐200a, to inhibit miR‐200a activity. miR‐200a regulates Wnt and TGFß signaling and inhibition of miR‐200a allows for cells to convert to a progeny phenotype in the presence of PAX9. These cells express early dental mesenchyme and stem cell markers making them suitable for tooth regeneration therapies. Current experiments are using reprogrammed dental epithelial and mesenchyme cells to form a tooth organ. These data demonstrate a new method for cell reprogramming and tissue regeneration.Support or Funding InformationSupport for this research was provided from grant DE013941 and DE018885 from the National Institute of Dental and Craniofacial Research.
Temporal Distribution of EPAC‐1 and EPAC‐2 on Normal Skin and Cutaneous Wound Healing of Chronically Stressed Mice
Authors: Romana‐Souza Bruna , dos Santos Jeanine Salles , Schmidt Martina , Monte‐Alto‐Costa Andréa , Porto Luis Cristóvão 
Published: November 2023

Psychological stress‐induced catecholamines impair cutaneous wound healing through engagement of different pathways. Recently, it was demonstrated that beta‐adrenoreceptor driven generation of cyclic adenosine monophosphate (cAMP) signals via adenylyl cyclase and exchange protein directly activated by cAMP (EPAC) in skin wound healing. However, it has not been shown yet if chronic stress‐induced high catecholamine levels could alter the expression of EPAC‐1 and EPAC‐1 in cutaneous wound healing. Thus, our aim was to observe the temporal distribution of the EPAC‐1 and EPAC‐2 in normal skin and full‐thickness excisional wounds of chronically stressed mice. Adult male mice (n=15) were submitted to a rotational stress which psychological stress model applied in rodents. For this, animals were daily spun at 115 rpm for 15 min every hour during 24 h, until euthanasia. Other animals (n=15) did not submit to stressful environment. Three days after stress protocol, two full‐thickness cutaneous lesions (8‐mm diameter) were created in the animals' dorsum. The wound contraction was macroscopically measured. Lesions were collected 3, 7 and 10 days after wounding. This study was approved by the Ethical Committee for Animal Use of the State University of Rio de Janeiro (n° 027/2014). The macroscopic analysis showed that chronic stress delayed wound contraction when compared to control group 3, 7 and 10 days after wounding. The protein EPAC‐1 expression was greater in the normal skin of stressed mice than in the normal skin of non‐stressed mice. In the wound area, protein EPAC‐1 expression was diminished stress group 7 and 10 days after wounding when compared to control group. The protein EPAC‐2 expression was decreased in the normal skin and wound area of stressed mice after 7 and 10 days when compared to non‐stressed mice. In conclusion, stress alters the protein expression of EPAC‐1 and EPAC‐2 on normal skin and cutaneous wound healing of mice.Support or Funding InformationThe Coordination for the Improvement of Higher Education Personnel (CAPES) and the National Council of Research (CNPq)
Regeneration of Rats Mental Nerve with Mesenchymal Stem Cells Obtained from Dental Pulp of Human Exfoliated Deciduous Teeth
Authors: Sasaki Robson Tetsuo , Saez Daniel Martinez , Nicoliche Tiago , Monteiro Bábyla Geraldes , Almeida Sonia Regina Yokomizo , Kerkis Irina , Silva Marcelo Cavenaghi Pereira 
Published: November 2023

The aim of this study was to evaluate the regeneration of mental nerve after injury by compression and treatment with mesenchymal cells obtained from dental pulp of human exfoliated deciduous teeth (SHED). Dental pulp from human exfoliated deciduous teeth were obtained from children aged 5–7 years old. Thirty‐six male Wistar rats, divided into 3 study groups (n=12): Positive Control (PC) – no lesion, Negative Control (NC) ‐ lesion by compression; and Experimental (SHED) ‐ lesion by compression and treatment with SHED. After the compression of the nerves, groups NC and SHED received application of media culture and media culture associated with SHED, respectively. Euthanasia was performed at 3, 7, 14 and 21 days for immunohistochemistry and transmission electron microscopy analysis. The immunohistochemical reaction was performed for nerve growth factor (NGF) detection. Ultra‐thin sections with 0.3μm were contrasted and observed under transmission electron microscope with 40,000× and 100,000× magnifications. NGF expression was present in all groups and time span studies, but less expressed in group NC while the staining was stronger in group SHED at 21 days. The ultrastructural analysis showed after 14 days of treatment the group SHED showed similar morphological aspects when compared with the PC group. It can be concluded that the use of mesenchymal cells obtained from dental pulp of human exfoliated deciduous teeth for the regeneration of rats mental nerve is efficient in a single dose of application after two weeks of treatment.Support or Funding InformationCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Premature ovarian failure patient‐specific induced pluripotent stem (iPS) cells show compromised potential to develop to germ cells and granulosa cells
Authors: Lu Chen‐Yu , Chen Yu‐An , Wu Fang‐Chun , Ho Hong‐Nerng , Chen Hsin‐Fu 
Published: November 2023

Premature ovarian failure (POF; also called primary ovarian insufficiency, POI), is a common endocrine disorder in females, accounting for 1–2% in reproductive‐age women. Patients with the disease experience spontaneous menopause before the age of 40 years, and frequently encounter significant difficulties in overcoming the problems of infertility and menopausal syndrome. However, the mechanisms underlying POF are difficult to study due to the lack of a suitable disease model. Recently pluripotent stem cells, including human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells, are becoming useful cell sources to study and dissect the molecular mechanisms underlying in vitro differentiation into specialized cell types. We and other researchers have previously shown that pluripotent stem cells including both ES cells and iPS cells are capable of developing into primordial germ cells (PGCs) and ovarian granulosa‐like cells. In this study therefore, we aimed to derive human iPS cells from POF patients and further differentiate the stem cells to germ cell‐related lineages. Now we have successfully derived and characterize patient‐specific iPS cell lines from five POF patients and two control subjects by transduction of skin fibroblasts or peripheral blood mononuclear cells (PBMCs) with four defined factors (OCT3/4, SOX2, KLF4 and c‐Myc) using Sendai virus reprogramming method. The resultant human iPS cells were then induced by using the cytokine cocktails towards PGCs, which expressed germ cell markers VASA, GDF9 and meiosis marker SCP3, and also towards functional granulosa‐like cells, which expressed FOXL2, FSHR, AMHR2, AMH and CYP19A1. We also observed that the POF‐patient iPS cells derivatives showed differential and reduced expression levels of the granulosa cell markers (AMH, AMHR2, CYP19A1 etc.) and germ cell marker VASA, when compared to control iPS and ES cells. The present data thus supported that we could produce POF‐specific iPS cells and induce the stem cells in vitro to PGCs and granulosa cells. Considering the altered phenotypes of the stem cell derivatives, the POF patient‐specific iPS cells derived in this study thus represent a potentially useful disease model for further studies into the disease mechanisms and to screen drugs that may be useful for treating and arresting POI.Support or Funding InformationNational Science Council of the Republic of China (NSC) and National Taiwan University Hospital (NTUH), Taiwan
Molecular Mechanisms of Mutagenesis Induced by DNA Repair
Authors: Shen Birong , Chapman Joseph , Furano Anthony V. 
Published: November 2023

Cancer is a complicated outcome of multiple somatic mutations, however, the molecular mechanisms underlying most of the mutations are unknown. Previous work in our lab using a shuttle vector system showed that repair of mismatched base pairs and other lesions induced mutations in flanking DNA. Most of these mutations exhibited a mutational spectrum typical of a cytidine deaminase, APOBEC3B (A3B). We also showed that both base excision repair and mismatch repair (BER & MMR) are required and the amount of APOBEC bound to the shuttle vector is proportionate to the extent of mutagenesis. We therefore postulated that the single‐stranded DNA substrate of the APOBEC deaminase can be generated during repair (Chen et al eLife 2014;3:e02001).Recent studies reported the involvement of A3B in generating mutations in some cancers, an example of the so‐called “mutator phenotype” long‐known to typify many cancers. We are currently using our shuttle vector to elucidate the mechanism of APOBEC‐mediated mutations in breast cancer cell lines. We found that repair of pre‐formed mismatches inserted into the shuttle vector can induce APOBEC‐mediated mutations, indicating that this system can monitor and reflect the mutational environment in tumor cells. We showed that two cancer cell lines, which contained comparable amounts of A3B, exhibited dramatically different repair‐induced mutation rates. Therefore, we postulated that the mutation rate is related to the amount of the repair‐generated APOBEC single‐stranded DNA substrate in each cell line, perhaps due to dysregulation of their DNA repair enzymes. Quantification of these enzymes showed that the relative level of some BER genes was positively correlated with repair‐induced mutagenesis, and preliminary results showed that knock down of one such gene reduced mutagenesis. Thus, our shuttle vector system could provide a tractable experimental system to explore the mechanism of the cancer cell mutator phenotype.