Miracle-dz/UniFAR-FTUs huggingface dataset:
Soy protein isolate down‐regulates caveolin‐1 expression to suppress osteoblastic cell senescence pathways

It has been suggested that the beneficial effects of soy protein isolate (SPI) on bone quality are due to either stimulation of estrogenic signaling via isoflavones or through a novel and as yet uncharacterized nonestrogenic pathway. In our study, SPI‐fed rat serum inhibited the osteoblastic cell senescence pathway. This effect was accompanied by stimulation of cell differentiation, proliferation, and significant restoration of replicative senescent bone marrow mesenchymal ST2 cells (passaged 30 times). These effects were reproduced in bone from 5‐wk‐old intact and 10‐wk‐old ovariectomized female rats fed SPI diets. Caveolin‐1 and p53 expression was decreased in bone in SPI‐fed, but not in 17β‐estradiol (E2)‐treated rats. In cell culture studies, membranous caveolin‐1 and nuclear p53 expression was greater in replicative senescent ST2 cell cultures than in earlier passaged cells. SPI‐fed rat serum significantly down‐regulated both caveolin‐1 and p53 in senescent and nonsenescent cells. Replicative senescent ST2 cells exhibited a strong association among caveolin‐1, p53, and mouse double minute 2 homologue (mdm2), which was inhibited by SPI‐fed rat serum. Overexpression of caveolin‐1 in ST2 cells resulted in increased expression of p53 and p21, whereas, knockdown of caveolin‐1 using shRNA led to increases in mdm2 and eliminated SPI‐fed rat serum's effects on p53 and p21 expression. In contrast, manipulation of caveolin‐1 expression did not affect the actions of E2 or isoflavones on p53 expression in either ST2 or OB6 cells. These results suggest that caveolin‐1 is a mediator of nonestrogenic SPI effects on bone cells. —Zhang, J., Lazarenko, O. P., Blackburn, M. L. , Badger, T. M. , Ronis, M. J. J., Chen, J.‐R. Soy protein isolate down‐regulates caveolin‐1 expression to suppress osteoblastic cell senescence pathways. FASEB J. 28, 3134–3145 (2014). www.fasebj. org
Enteric nervous system abnormalities in inflammatory bowel diseases

Abstract Various studies have described abnormalities of the enteric nervous system (ENS) in tissue samples from patients with chronic idiopathic inflammatory bowel diseases (IBD). The distribution of density of the different cell types of the ENS was however not studied in a systematic way. The aim of this study was to examine the density of neurons, enteroglial cells and interstitial cells of Cajal (ICC) in the different plexuses of the ENS in samples from patients with Crohn’s disease (CD), ulcerative colitis (UC) and controls. Tissue samples from 16 patients with CD (ileum) and 16 patients with UC obtained in involved and non‐involved areas were studied using immunohistochemistry with antibodies directed against neuron‐specific enolase, S100, C‐Kit and CD3. Sections were analysed blindly by two pathologists and the number of positive cells was counted for each type. Overall, an increase was noted for neuronal cell bodies, enteroglia and ICC in the deep muscular plexus in CD. In uninvolved areas of CD patients, the number of enteroglial cells was decreased. In UC, an increase of ICC in the muscularis propria and enteroglial cells was observed in diseased tissue. The study confirms the presence of abnormalities of the different cells of the ENS in IBD. The presence of lesions in samples from uninvolved areas, such as a reduction of enteroglia, supports a pathogenetic role of the ENS.
Structural Basis for Substrate Recognition and Hydrolysis by Mouse Carnosinase CN2*

l-Carnosine is a bioactive dipeptide (β-alanyl-l-histidine) present in mammalian tissues, including the central nervous system, and has potential neuroprotective and neurotransmitter functions. In mammals, two types of l-carnosine-hydrolyzing enzymes (CN1 and CN2) have been cloned thus far, and they have been classified as metallopeptidases of the M20 family. The enzymatic activity of CN2 requires Mn2+, and CN2 is inhibited by a nonhydrolyzable substrate analog, bestatin. Here, we present the crystal structures of mouse CN2 complexed with bestatin together with Zn2+ at a resolution of 1.7Å and that with Mn2+ at 2.3Å. CN2 is a homodimer in a noncrystallographic asymmetric unit, and the Mn2+ and Zn2+ complexes closely resemble each other in the overall structure. Each subunit is composed of two domains: domain A, which is complexed with bestatin and two metal ions, and domain B, which provides the major interface for dimer formation. The bestatin molecule bound to domain A interacts with several residues of domain B of the other subunit, and these interactions are likely to be essential for enzyme activity. Since the bestatin molecule is not accessible to the bulk water, substrate binding would require conformational flexibility between domains A and B. The active site structure and substrate-binding model provide a structural basis for the enzymatic activity and substrate specificity of CN2 and related enzymes.
Design of treatment trials in irritable bowel syndrome: opioid agonists and atypical benzodiazepine antagonists

Abstract Irritable bowel syndrome (IBS) is a common gastrointestinal disorder characterized by abdominal pain and altered bowel function. Few agents are available for the treatment of IBS. Historically, one impediment to the development of agents to treat IBS was lack of a uniform and robust clinical trial design. Studies occurred with different durations of treatment, endpoints and with different target populations. Great advances have been made over the past decade in trial design including: optimal duration of study, mode of data collection, populations to evaluate and identification of endpoints. Using these refinements, it was possible to demonstrate the efficacy of some new agents. These advances are illustrated by a review of trials with kappa opioid agonists and atypical benzodiazepine antagonists, which appear to be promising new classes of treatments for symptoms in IBS.
Tauopathy contributes to synaptic and cognitive deficits in a murine model for Alzheimer's disease

Tau alterations are now considered an executor of neuronal demise and cognitive dysfunction in Alzheimer's disease (AD). Mouse models combining amyloidosis and tauopathy and their parental counterparts are important tools to further investigate the interplay of abnormal amyloid‐β (Aβ) and Tau species in pathogenesis, synaptic and neuronal dysfunction, and cognitive decline. Here, we crossed APP/PS1 mice with 5 early‐onset familial AD mutations (5xFAD) and TauP301S (PS19) transgenic mice, denoted F+/T+ mice, and phenotypically compared them to their respective parental strains, denoted F+/T– and F–/T+ respectively, as controls. We found dramatically aggravated tauopathy (~10‐fold) in F+/T+ mice compared to the parental F–/T+ mice. In contrast, amyloidosis was unaltered compared to the parental F+/T– mice. Tauopathy was invariably and very robustly aggravated in hippocampal and cortical brain regions. Most important, F+/T+ displayed aggravated cognitive deficits in a hippocampus‐dependent spatial navigation task, compared to the parental F+/T– strain, while parental F–/T+ mice did not display cognitive impairment. Basal synaptic transmission was impaired in F+/T+ mice compared to nontransgenic mice and the parental strains (≥40%). Finally, F+/T+ mice displayed a significant hippocampal atrophy (~20%) compared to nontransgenic mice, in contrast to the parental strains. Our data indicate for the first time that pathological Aβ species (or APP/PS1) induced changes in Tau contribute to cognitive deficits correlating with synaptic deficits and hippocampal atrophy in an AD model. Our data lend support to the amyloid cascade hypothesis with a role of pathological Aβ species as initiator and pathological Tau species as executor. —Stancu, I. ‐C. , Ris, L. , Vasconcelos, B. , Marinangeli, C. , Goeminne, L. , Laporte, V. , Haylani, L. E. , Couturier, J., Schakman, O. , Gailly, P., Pierrot, N. , Kienlen‐Campard, P., Octave, J.‐N. , Dewachter, I. Tauopathy contributes to synaptic and cognitive deficits in a murine model for Alzheimer's disease. FASEB J. 28, 2620–2631 (2014). www.fasebj. org
Tuberculosis antigen‐induced expression of IFN‐α in tuberculosis patients inhibits production of IL‐1β

The mechanism by which IFN‐α regulates the host response to Mycobacterium tuberculosis (M.tb) infection in humans is poorly understood. In the present study, we found that freshly isolated pleural fluid mononuclear cells (PFMCs) from tuberculous pleural effusion but not peripheral blood mononuclear cells (PBMCs) spontaneously expressed IFN‐α and IL‐1β in vivo. In addition, exogenous IFN‐α significantly inhibited production of IL‐1β in PFMCs after stimulation with Bacillus Calmette‐Guérin (BCG). To further evaluate the effect of endogenous IFN‐α on BCG‐induced IL‐1β production, a neutralizing antibody to IFN‐α was added to the cultures of BCG‐stimulated PFMCs. As expected, neutralization of IFN‐α by antibody significantly enhanced the production of IL‐1β. Notably, we showed that IFN‐α inhibited production of IL‐1β through 2 distinct mechanisms: IFN‐α signaling, via the STAT1 transcription factor, suppressed caspase‐1‐dependent IL‐1β maturation, and IFN‐α induced the production of IL‐10 in a STAT1‐dependent manner in which IL‐10 reduced the abundance of IL‐1β. In contrast, we found that IFN‐α enhanced the production of IFN‐γ, and IFN‐γ also suppressed IL‐1β production in the PFMCs during BCG stimulation. Our findings demonstrate that IFN‐α employs distinct pathways for regulating IL‐1β production and reveal that in the case of M.tb infection, the induction of IFN‐α and IFN‐γ might be associated with M.tb immune escape and disease progression in infected humans. —Ma, J., Yang, B. , Yu, S. , Zhang, Y. , Zhang, X. , Lao, S. , Chen, X. , Li, B. , Wu, C. Tuberculosis antigen‐induced expression of IFN‐α in tuberculosis patients inhibits production of IL‐1β. FASEB J. 28, 3238–3248 (2014). www.fasebj. org
Rapid actin‐cytoskeleton–dependent recruitment of plasma membrane–derived dysferlin at wounds is critical for muscle membrane repair

Deficits in membrane repair may contribute to disease progression in dysferlin‐deficient muscular dystrophy. Dysferlin, a type‐II transmembrane phospholipid‐binding protein, is hypothesized to regulate fusion of repair vesicles with the sarcolemma to facilitate membrane repair, but the dysferlin‐containing compartments involved in membrane repair and the mechanism by which these compartments contribute to resealing are unclear. A dysferlin‐pHluorin [dysf‐pH‐sensitive green fluorescent protein (pHGFP)] muscle‐specific transgenic mouse was developed to examine the dynamic behavior and subcellular localization of dysferlin during membrane repair in adult skeletal muscle fibers. Live‐cell confocal microscopy of uninjured adult dysf‐pHGFP muscle fibers revealed that dysferlin is highly enriched in the sarcolemma and transverse tubules. Laser‐wounding induced rapid recruitment of ~30 μm of local dysferlin‐containing sarcolemma, leading to formation of stable dysferlin accumulations surrounding lesions, endocytosis of dysferlin, and formation of large cytoplasmic vesicles from distal regions of the fiber. Disruption of the actin cytoskeleton decreased recruitment of sarcolemma‐derived dysferlin to lesions in dysf‐pHGFP fibers without affecting endocytosis and impaired membrane resealing in wild‐type fibers, similar to findings in dysferlin deficiency (a 2‐fold increase in FM1‐43 uptake). Our data support a new mechanism whereby recruitment of sarcolemma‐derived dysferlin creates an active zone of high lipid‐binding activity at wounds to interact with repair vesicles and facilitate membrane resealing in skeletal muscle. —McDade, J. R. , Archambeau, A. , Michele, D. E. Rapid actin‐cytoskeleton‐dependent recruitment of plasma membrane‐derived dysferlin at wounds is critical for muscle membrane repair. FASEB J. 28, 3660–3670 (2014). www.fasebj. org
Intracrine sex steroid synthesis and signaling in human epidermal keratinocytes and dermal fibroblasts

Peripheral intracrine sex steroid synthesis from adrenal precursors dehydroepiandrosterone (DHEA) and DHEA‐sulfate has evolved in humans. We sought to establish if there are differences in intracrine, paracrine, and endocrine regulation of sex steroids by primary cultures of human skin epidermal keratinocytes and dermal fibroblasts. Microarray analysis identified multifunctional genes modulated by steroids, quantitative RT‐PCR (qRT‐PCR) mRNA expression, enzymatic assay aromatase activity, scratch assay cell migration, immunocytochemistry α‐smooth muscle actin (a‐SMA), and collagen gel fibroblast contraction. All steroidogenic components were present, although only keratinocytes expressed the organic anion organic anion transporter protein (OATP) 2B1 transporter. Both expressed the G‐protein‐coupled estrogen receptor (GPER1). Steroids modulated multifunctional genes, up‐regulating genes important in repair and aging [angiopoietin‐like 4 (ANGPTL4), chemokine (C‐X‐C motif) ligand 1 (CXCL1), lamin B1 (LMNB1), and thioredoxin interacting protein (TXNIP)]. DHEA‐sulfate (DHEA‐S), DHEA, and 17β‐estradiol stimulated keratinocyte and fibroblast migration at early (4 h) and late (24‐48 h) time points, suggesting involvement of genomic and nongenomic signaling. Migration was blocked by aromatase and steroid sulfatase (STS) inhibitors confirming intracrine synthesis to estrogen. Testosterone had little effect, implying it is not an intermediate. Steroids stimulated fibroblast contraction but not α‐SMA expression. Mechanical wounding reduced fibroblast aromatase activity but increased keratinocyte activity, amplifying the bioavailability of intracellular estrogen. Cultured fibroblasts and keratinocytes provide a biologically relevant model system to investigate the complex pathways of sex steroid intracrinology in human skin. —Pomari, E. , Valle, L. D. , Pertile, P., Colombo, L. , and Thornton, M. J. Intracrine sex steroid synthesis and signaling in human epidermal keratinocytes and dermal fibroblasts. FASEB J. 29, 508‐524 (2015). www.fasebj. org
Multimodality serial follow-up of thoracic aortic aneurysms

We identified 520 diagnostic imaging tests (MRI, CT, ECHO) performed in 205 patients (79 female 126 male) (age 20–94, mean 63.9) being followed at Yale-New Haven hospital for progression in size of their thoracic aortas. Estimated growth rates did not differ significantly across imaging modalities.The average maximal size of the thoracic aorta was 5.3 cm (range 3.5–10.0). Mean maximal size was 5.3 cm in the 326 imaging studies without dissections. Mean size at time of dissection (N=22) was 6.2 cm, and a time of rupture (N=7), 6.5 cm.Survival at 3 and 5 years was 66% and 45%, respectively. Subjects having aortic dissection had lower survival (58% 3 year; 15% 5 year) than the non-dissected cohort (70% 3 year; 60% 5 year). Mean estimated growth rate on 53 patients followed serially for aortic expansion was 0.1 cm/yr. Stepwise multivariate risk factor analysis indicated that large (>=6.0 cm) aneurysms grew significantly more rapidly (p<0.02), that aneurysms located in the ascending aorta or arch grew more slowly than aneurysms located in the descending thoracic or thoraco-abdominal aorta (p<0.04), and that aneurysms grew more rapidly among male patients than among females (p<0.02). We conclude that (1) the overall growth rate in thoracic aorta aneurysms is 0.1 cm/yr. (2) Large aneurysms grow more rapidly than smaller ones. (3) Aneurysms grow more slowly in the ascending thoracic aorta or arch than on other locations of the thoracic aorta. (4) Aneurysms appear to grow somewhat more rapidly among male patients than among females. (5) One-half of aortic dissections and ruptures occur at sizes <6.0 cm. (6) Long-term survival prospects are less for patients having dissected aortas. (7) 5.0–5.5 cm is an appropriate criterion for surgical intervention in the thoracic aorta to prevent rupture or dissection.
EphrinB2 signaling in osteoblasts promotes bone mineralization by preventing apoptosis

Cells that form bone (osteoblasts) express both ephrinB2 and EphB4, and previous work has shown that pharmacological inhibition of the eph‐rinB2/EphB4 interaction impairs osteoblast differentiation in vitro and in vivo. The purpose of this study was to determine the role of ephrinB2 signaling in the osteoblast lineage in the process of bone formation. Cultured osteoblasts from mice with osteoblast‐specific ablation of ephrinB2 showed delayed expression of osteoblast differentiation markers, a finding that was reproduced by ephrinB2, but not EphB4, RNA interference. Microcomputed tomography, histomorphometry, and mechanical testing of the mice lacking ephrinB2 in osteoblasts revealed a 2‐fold delay in bone mineralization, a significant reduction in bone stiffness, and a 50% reduction in osteoblast differentiation induced by anabolic parathyroid hormone (PTH) treatment, compared to littermate sex‐ and age‐matched controls. These defects were associated with significantly lower mRNA levels of late osteoblast differentiation markers and greater levels of osteoblast and osteocyte apoptosis, indicated by TUNEL staining and transmission electron microscopy of bone samples, and a 2‐fold increase in annexin V staining and 7‐fold increase in caspase 8 activation in cultured ephrinB2 deficient osteoblasts. We conclude that osteoblast differentiation and bone strength are maintained by antiapoptotic actions of ephrinB2 signaling within the osteoblast lineage. —Tonna, S. , Takyar, F. M. , Vrahnas, C. , Crimeen‐Irwin, B. , Ho, P. W. M. , Poulton, I. J., Brennan, H. J., McGregor, N. E. , Allan, E. H. , Nguyen, H. , Forwood, M. R. , Tatarczuch, L. , Mackie, E. J., Martin, T. J., Sims, N. A. , EphrinB2 signaling in osteoblasts promotes bone mineralization by preventing apoptosis. FASEB J. 28, 4482–4496 (2014). www.fasebj. org
A pivotal involvement of IFN‐' in the pathogenesis of acetaminophen‐induced acute liver injury

In wild‐type BALB/c mice, i.p. administration of acetaminophen (APAP; 750 mg/kg) induced intrahepatic IFN‐' mRNA expression and a marked increase in serum transaminase levels, leading to acute lethality of ~45%. Histopathological examination showed centrilobular hepatic necrosis with leukocyte infiltration and a large number of apoptotic hepatocytes 10 and 24 h after APAP challenge. mRNA expression of intercellular adhesion molecule 1, vascular cell adhesion molecule 1, interleukin (IL) 1a, IL‐1P, IL‐6, tumor necrosis factor a, monocyte chemoattractant protein 1, macrophage inflammatory protein (MIP) 1a, MIP‐2, KC, IP‐10, Mig, Fas, and inducible nitric oxide synthase was enhanced in the liver of wild‐type mice injected with APAP. To clarify the role of IFN‐' in this process, IFN‐'‐deficient mice were treated in the same manner. All IFN‐'‐deficient mice survived with reduced serum transaminase elevation and attenuated hepatic necrosis, leukocyte infiltration, and hepatocyte apoptosis. The gene expression of all molecules was significantly attenuated in IFN‐'‐deficient mice. Administration of an anti‐IFN‐' neutralizing antibody even 2 or 8 h after APAP challenge to wild‐type mice alleviated APAP‐induced liver injury, and all mice survived. Thus, IFN‐' is responsible for APAP‐induced liver injury by mediating leukocyte infiltration, hepatocyte apoptosis, and NO production as well as cytokine and chemokine production. Moreover, immunoneutralization of IFN‐' may be therapeutically effective for developing APAP‐induced liver injury. —Ishida, Y. , Kondo, T. , Ohshima, T. , Fujiwara, H. , Iwakura, Y. , Mukaida, N. A pivotal involvement of IFN‐' in the pathogenesis of acetaminophen‐induced acute liver injury. FASEB J. 16, 1227–1236 (2002)
AUTONOMIC CONTROL OF SUBMANDIBULAR CHROMOGRANIN A SECRETION IN THE ANAESTHETIZED RAT

Region-specific radioimmunoassay (RIA) developed with anti-rat chromogranin A (CgA) (359-389) serum was used for estimating levels of CgA-like immunoreactivity (CgA-like IR) in saliva and plasma of rats. Under pentobarbitone anaesthesia, the sympathetic and parasympathetic nerves innervating the submandibular gland were stimulated in both efferent and afferrent direction. The stimulation of the nerves caused elevation of CgA-like IR level in the plasma from the resting level (0.97 i 0.34 nM) to 1.23 i 0.42 nM, the level in the saliva from non-collectable level to 631 fl: 359 nM, and salivary flow rate from non-collectable rate to l0.7i2. 6;,¢L min" g'1. The intramural injection of acetylcholine (total 0.2 mL of 0.1 mM solution) into the submandibular gland induced a distinct increase in flow rate of the saliva from non-collectable level to 16 i 5.5 ;1L min‘1 g", which contained 5,964 in 2,065 nM CgA-like IR. CgA-like IR was immunohistochemically ascertained in duct cells but not in neural elements and acinar cells in rat submandibular gland. These results are compatible with a novel view that the source of salivary CgA-like IR is the duct cells, which release highly concentrated CgA-like IR into saliva in response to activation of the afferent autonomic axis of the stress system, and provide experimental bases for a view that CgA-like IR may be a sensitive and substantial index for psychosomatic stress response. Protein and fluid secretion from rat submandibular glands have been studied in response to sympathetic and parasympathetic nerve stimulation in vivo (1, 2, 3, 6, 8, 9), and to autonomimetic agonists in vivo and in vz'z‘r0 (1, 4, 5, 10). In general term, sympathetic electric stimulation of nerves evokes a secretion of saliva low in volume but rich in protein, and causes an extensive exocytosis of both acinar and granular duct cells, whereas parasympathetic electrical stimulation of the nerves induces a copious flow of saliva low in protein (see 3 for reference). The secretion of Correspondence to: Dr T. Kanno at the above address. Tel: +81-544-22-2771; Fax: +81-544-22-2771 protein from the granular ducts is generally considered to be an a—adrenergic response, whereas that the acini involves 6’-adrenergic receptor activation (l, 7). Despite this seemingly orderly compartmentalization of autonomic function, several observations raise questions about the neural regulation of protein secretion in the rat submandibular gland. Anderson, Garrett, Zhang, Proctor and Shori (3) concluded that parasympathetic stimulation caused a moderate increase in the concentration and output of acinar peroxidase with increasing stimulation rates, but the concentration of kallikreins secreted from the granular ducts did not change significantly. They suspected that parasympathetic secretion of kallikreins was
Cdt1 and Cdc6 Are Destabilized by Rereplication-induced DNA Damage*

The replication factors Cdt1 and Cdc6 are essential for origin licensing, a prerequisite for DNA replication initiation. Mechanisms to ensure that metazoan origins initiate once per cell cycle include degradation of Cdt1 during S phase and inhibition of Cdt1 by the geminin protein. Geminin depletion or overexpression of Cdt1 or Cdc6 in human cells causes rereplication, a form of endogenous DNA damage. Rereplication induced by these manipulations is however uneven and incomplete, suggesting that one or more mechanisms restrain rereplication once it begins. We find that both Cdt1 and Cdc6 are degraded in geminin-depleted cells. We further show that Cdt1 degradation in cells that have rereplicated requires the PCNA binding site of Cdt1 and the Cul4DDB1 ubiquitin ligase, and Cdt1 can induce its own degradation when overproduced. Cdc6 degradation in geminin-depleted cells requires Huwe1, the ubiquitin ligase that regulates Cdc6 after DNA damage. Moreover, perturbations that specifically disrupt Cdt1 and Cdc6 degradation in response to DNA damage exacerbate rereplication when combined with geminin depletion, and this enhanced rereplication occurs in both human cells and in Drosophila melanogaster cells. We conclude that rereplication-associated DNA damage triggers Cdt1 and Cdc6 ubiquitination and destruction, and propose that this pathway represents an evolutionarily conserved mechanism that minimizes the extent of rereplication.