Miracle-dz/UniFAR-FTUs huggingface dataset:
Notochordal-cell derived extracellular vesicles exert regenerative effects on canine and human nucleus pulposus cells

During intervertebral disc ageing, chondrocyte-like cells (CLCs) replace notochordal cells (NCs). NCs have been shown to induce regenerative effects in CLCs. Since vesicles released by NCs may be responsible for these effects, we characterized NC-derived extracellular vesicles (EVs) and determined their effect on CLCs. EVs were purified from porcine NC-conditioned medium (NCCM) through size exclusion chromatography, ultracentrifugation or density gradient centrifugation. Additionally, the EVs were quantitatively analyzed by high-resolution flow cytometry. The effect of NCCM-derived EVs was studied on canine and human CLC micro-aggregates in vitro and compared with NCCM-derived proteins and unfractionated NCCM. Porcine NCCM contained a considerable amount of EVs. NCCM-derived EVs induced GAG deposition in canine CLCs to a comparable level as NCCM-derived proteins and unfractionated NCCM, and increased the DNA and glycosaminoglycan (GAG) content of human micro-aggregates, although to a lesser extent than unfractionated NCCM. The biological EV effects were not considerably influenced by ultracentrifugation compared with size exclusion-based purification. Upon ultracentrifugation, interfering GAGs, but not collagens, were lost. Nonetheless, collagen type I or II supplemented to CLCs in a concentration as present in NCCM induced no anabolic effects. Porcine NCCM-derived EVs exerted anabolic effects comparable to NCCM-derived proteins, while unfractionated NCCM was more potent in human CLCs. GAGs and collagens appeared not to mediate the regenerative EV effects. Thus, NC-derived EVs have regenerative potential, and their effects may be influenced by the proteins present in NCCM. The optimal combination of NC-secreted factors needs to be determined to fully exploit the regenerative potential of NC-based technology.
The investigation of glutathione peroxidase, lactoferrin, myeloperoxidase and interleukin-1beta in gingival crevicular fluid: implications for oxidative stress in human periodontal diseases.

BACKGROUND Human periodontal diseases are inflammatory disorders that are the result of complex interactions between periodontopathogens and the host's immune response. Two important and interrelated factors are involved in the pathophysiological progression of periodontal diseases, i.e. the activation of immune system and the production of oxygen radicals and their related metabolites. Increased production of oxygen radicals may contribute to oxidative stress, which is reported to be involved in many diseases, including periodontal diseases. OBJECTIVES The objective of this study was to investigate glutathione peroxidase, lactoferrin and myeloperoxidase, which play an essential role in free radical production and defenses, and the proinflammatory cytokine interleukin-1beta (IL-1beta), which is important in the regulation of immunological and inflammatory reactions in human periodontal diseases. METHODS Gingival crevicular fluid (GCF) samples were collected from 27 subjects, 19 periodontitis patients and eight healthy controls, ranging in ages from 24 to 62 years. Clinical parameters were recorded. GCF glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta were analyzed by enzyme-linked immunosorbent assays (ELISA). RESULTS The periodontitis sites exhibited significantly greater total amount of glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta than healthy sites. Total amount of glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta was positively correlated with plaque index, gingival index, probing depth and probing attachment level (p < 0.05). CONCLUSION The imbalance between the levels of myeloperoxidase/IL-1beta and glutathione peroxidase/lactoferrin could result in tissue damage of reactive oxygen species (ROS) in periodontitis which is initiated and perpetuated by the chronic insults of periodontopathogens.
Cellular Angiolipoma of the Breast: Immunohistochemical Study and Review of the Literature

We report a rare case of cellular angiolipoma of the breast in a 73‐year‐old woman with abnormal mammographic findings. Histopathologic examination of the tumor revealed cellular spindle cell areas associated with collapsed and patent blood vessels in a background of mature adipose tissue. A salient feature of this tumor, which was an important clue to the diagnosis, was the presence of intravascular fibrin thrombi. Immunohistochemical studies showed reactivities for endothelial and muscle markers, highlighting the obscured angiomatous component in the cellular areas of the tumor. Failure to recognize this entity may potentially lead to diagnostic pitfalls that include angiosarcoma and Kaposi's sarcoma, two of the better recognized entities in this region.
High-Affinity Interaction between Gram-Negative Flagellin and a Cell Surface Polypeptide Results in Human Monocyte Activation

ABSTRACT Flagella from diverse gram-negative bacteria induce tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) synthesis by human monocytes (F. Ciacci-Woolwine, P. F. McDermott, and S. B. Mizel, Infect. Immun. 67:5176–5185, 1999). In this study, we establish that purified flagellin (FliC or FljB), the major filament protein from Salmonella enterica serovar Enteritidis,S. enterica serovar Typhimurium, and Pseudomonas aeruginosa, is an extremely potent inducer of TNF-α production by human monocytes and THP-1 myelomonocytic cells. Fifty percent of maximal TNF-α production (EC50) was obtained with 1.5 × 10−11 M flagellin (0.75 ng/ml). Mutagenesis studies revealed that the central hypervariable region of flagellin is essential for the TNF-α-inducing activity of the protein. Although less active than the wild-type protein, a Salmonellaflagellin mutant composed of only the central hypervariable region retained substantial TNF-α-inducing activity at nanomolar concentrations. In contrast, the conserved amino- and carboxy-terminal regions are inactive. Mutational analysis of the hypervariable region revealed that it contains two equally active TNF-α-inducing domains. The ability of THP-1 cells to respond to purified flagellins is dramatically reduced by mild trypsin treatment of the cells. Taken together, our results demonstrate that the cytokine-inducing activity of flagellins from gram-negative bacteria results from the interaction of these proteins with high-affinity cell surface polypeptide receptors on monocytes.
Unsustained multiplication of treponema pallidum (nichols virulent strain) in vitro in the presence of oxygen

Treponema pallidum (Nichols virulent strain) was incubated with or without oxygen using a modified medium supplemented with reduced glutathione and a variety of nutrients (PRNF10-B). Two- to fourfold increases in treponemal numbers were observed in cultures without mammalian cells within 96 h of incubation under 5 to 6% oxygen. Treponemal motility and multiplication were maintained more satisfactorily in cultures that were diluted and transferred daily, using an equal volume of fresh medium. Treponemes incubated without oxygen did not significantly increase in number. Virulent microorganisms were detected for at least 96 h in the cell-free system. In the presence of 3 to 4% oxygen, two- to fivefold increases in treponemal numbers were observed in the supernatant fluids of cultures containing human prepuce cells after 48 to 120 h at 35 degrees C. Without oxygen, treponemal numbers rarely approached a threefold increase. Virulent treponemes were detected by the rabbit skin lesion test after at least 120 h in vitro. Regardless of the system of incubation, increases in treponemal numbers could not be sustained for longer than 120 h, and treponemal virulence decreased as a function of time in vitro.
Pharmacodynamics of Abacavir in an In Vitro Hollow-Fiber Model System

ABSTRACT Abacavir is a potent new carbocyclic nucleoside analogue. We employed our hollow-fiber pharmacodynamic modeling system to examine the antiretroviral effects of different abacavir exposures, as well as the impact of the schedule of drug administration on efficacy. Dose ranging of abacavir revealed that a concentration of four times the 50% effective concentration (EC50) (approximately the EC95) was required to inhibit the replication of human immunodeficiency virus type 1 (HIV-1) (strain MN) either in a continuous-infusion hollow-fiber experiment or in a classical tissue culture flask experiment. In contrast to earlier work with another drug class (HIV-1 protease inhibitors), addition of physiological amounts of the human drug binding proteins albumin and α1 acid glycoprotein revealed that there was little impact on the antiviral effect of the drug. Comparison of equivalent exposures (an area under the concentration-time curve [AUC] developed by approximately 500 mg per day of orally administered abacavir), either in a continuous-infusion mode or as a single oral dose of abacavir, demonstrated no difference in the ability to suppress either strain IIIB or strain MN. Comparison of administration of 250 mg every 12 h (q12h) versus once-daily administration of 500 mg for strain MN again showed no significant difference in suppressive effect. These experiments were carried out over 8 to 15 days. Because of these promising initial results, we extended the experiment to 30 days and examined three different schedules of administration that generated the same AUC at 24 h (AUC24): 300 mg q12h, 600 mg q24h, and 1,200 mg q48h. The aim of the last of these regimens was to definitively demonstrate schedule failure. There was little difference between the 1,200-mg q48h treatment group and the untreated control at 30 days. Likewise, there was little difference between the 600-mg q24h and 300-mg q12h treatment groups. However, at circa day 18 of the experiment, there was a small increase in viral output of p24 in the once-daily dosing unit. Examination of virus from all groups demonstrated no phenotypic or genotypic differences. The small difference in hollow-fiber unit p24 in the once-daily dosing group was not due to emergence of resistance over the 30-day single-drug exposure. We conclude that the dose of abacavir currently being studied in clinical trials (300 mg orally q12h) will be efficacious for the majority of sensitive clinical isolates of HIV-1. These in vitro data also suggest that this drug may be able to be administered to patients on a once-daily basis at a dose of 600 mg.
Influence of the mesenchymal cell source on oral epithelial development

The extent of the influence of mesenchymal tissue on epithelial development is still debated, and elucidation of epithelial–mesenchymal interactions should be of relevance for controlling normal as well as pathological growth and development. The aim of the present study was to elucidate the influence of the mesenchymal cell type on oral mucosa epithelial development in vitro, using tissue‐engineering principles, by including three different sources for mesenchymal cell type, viz. oral mucosa, skin and cornea, each of them presenting a distinct type of epithelium in situ. We investigated epithelial–mesenchymal interactions, considering both morphological criteria and protein expression (filaggrin, keratin 10, keratin 12, keratin 13 and laminin 5). The results of the histology, immunohistochemistry and transmission electron microscopy of the three types of tissue‐engineered constructs composed of mesenchymal cells of different sources (oral, dermal and corneal fibroblasts) and of the same oral epithelial cells showed that the mesenchymal cell source had a significant influence on the thickness and ultrastructure of the epithelium, but not on the differentiation of oral epithelial cells, which might be an intrinsic property of these cells due to their genetic programming. Copyright © 2011 John Wiley & Sons, Ltd.
Kinases and phosphatases and tau sites involved in Alzheimer neurofibrillary degeneration

Microtubule associated protein (MAP) tau is abnormally hyperphosphorylated in Alzheimer's disease (AD) and related tauopathies; in this form it is the major protein subunit of paired helical filaments (PHF)/neurofibrillary tangles. However, the nature of protein kinases and phosphatases and tau sites involved in this lesion has been elusive. We investigated self‐assembly and microtubule assembly promoting activities of hyperphosphorylated tau isolated from Alzheimer disease brain cytosol, the AD abnormally hyperphosphorylated tau (AD P‐tau) before and after dephosphorylation by phosphoseryl/phosphothreonyl protein phosphatase‐2A (PP‐2A), and then rephosphorylation by cyclic AMP‐dependent protein kinase (PKA), calcium, calmodulin‐dependent protein kinase II (CaMKII), glycogen synthase kinase‐3β (GSK‐3β) and cyclin‐dependent protein kinase 5 (cdk5) in different kinase combinations. We found that (i) dephosphorylation of AD P‐tau by PP‐2A inhibits its polymerization into PHF/straight filaments (SF) and restores its binding and ability to promote assembly of tubulin into microtubules; (ii) rephosphorylation of PP‐2A‐dephosphorylated AD P‐tau by sequential phosphorylation by PKA, CaMKII and GSK‐3β or cdk5, and as well as by cdk5 and GSK‐3β, promotes its self‐assembly into tangles of PHF similar to those seen in Alzheimer brain, and (iii) phosphorylation of tau sites required for this pathology are Thr231 and Ser262, along with several sites flanking the microtubule binding repeat region. Phosphorylation of recombinant human brain tau441 yielded similar results as the PP‐2A dephosphorylated AD P‐tau, except that mostly SF were formed. The conditions for the abnormal hyperphosphorylation of tau that promoted its self‐assembly also induced the microtubule assembly inhibitory activity. These findings suggest that activation of PP‐2A or inhibition of either both GSK‐3β and cdk5 or one of these two kinases plus PKA or CaMKII might be required to inhibit Alzheimer neurofibrillary degeneration.
Iron availability of a fortified processed wheat cereal: a comparison of fourteen iron forms using an in vitro digestion/human colonic adenocarcinoma (CaCo-2) cell model

In this three-phase study we first compared the availability of fourteen Fe forms in a wheat-based ready-to-eat breakfast cereal using an in vitro digestion/human colonic adenocarcinoma (CaCo-2) cell model. We then investigated the effect of milk and/or coffee on those fortified cereals found in phase 1 to show promising increases in Fe availability. The Fe forms assessed in phase 1 were reduced (control), carbonyl, electrolytic, FePO4, FeSO4, FeCO3, Na2FeEDTA, Ferrochel® (Albion Laboratories, Clearfield, UT, USA; ferrous bis-glycinate), encapsulated ferrous fumarate, FeSO4, ferrous lactate and Biofer® (LipoTech, Britwell Salome, Oxfordshire; FeSO4), SQM® (Sea-Questra-Min Iron; Quali Tech, Chaska, MN, USA; polysaccharide-complexed FeSO4) and Sun Active® (Taiyo Kagaku, Yokkaichi, Japan). All these forms increased Fe uptake compared with the unfortified cereal. Relative to the control, the following increases in Fe availability were observed: electrolytic, 52 %; ferrous fumarate, 30–35 %; Sun Active, 78 %; Ferrochel, 125 %; Na2FeEDTA, 291 %. Recent human studies have shown similar data with regard to Ferrochel, FeSO4 and Na2FeEDTA, with the latter being more bioavailable. Our phase-2 studies indicated that the addition of milk to FeSO4-fortified cereal increased Fe availability, but this availability was markedly decreased by the addition of coffee to the digest. Conversely, a loss in availability from Na2FeEDTA was observed with the addition of milk; however, the addition of coffee did not markedly affect Fe availability from this form. In phase-3 studies we observed increased Fe availability upon the addition of milk to cereals containing Ferrochel, FeSO4, Sun Active, a mixture of reduced Fe and Na2FeEDTA or reduced Fe. For these forms we did not assess the behaviour after the addition of coffee. In conclusion, when considering possible fortificants for optimizing Fe bioavailability within a foodstuff, it is of paramount importance to consider the interaction between the fortified foodstuffs and other components of the meal (such as milk and coffee with a breakfast).
Inflexinol Reduces Severity of Acute Pancreatitis by Inhibiting Nuclear Factor-&kgr;B Activation in Cerulein-Induced Pancreatitis

Objectives To examine the effect of inflexinol on the development of acute pancreatitis (AP) and to investigate the mechanisms responsible for the protective effect against AP. Methods Acute pancreatitis was induced in mice by intraperitoneal injection of cerulein. Inflexinol was administered intraperitoneally 4 times every 6 hours from 1 hour before the first cerulein injection. Serum amylase activity and histology of the pancreas were measured. Determination of pancreatic nuclear factor-&kgr;B (NF-&kgr;B) p65 expression was conducted by Western blotting and immunohistochemistry to investigate the mechanisms responsible for the inflexinol effects. Results Serum amylase activity in the cerulein group was significantly higher than that in the control group (P < 0.05). Pancreatic histology revealed marked inflammatory changes in the cerulein group such as interstitial edema, vacuolization, necrosis, and infiltration of inflammatory cells; and Western blotting and immunohistochemistry showed marked NF-&kgr;B p65 expression. Treatment with inflexinol significantly attenuated the inflammatory changes in pancreatic histology at 24, 48, and 72 hours (P < 0.05). Pancreatic NF-&kgr;B p65 expression decreased significantly after inflexinol treatment (P < 0.05). Conclusion Inflexinol reduced the severity of cerulein-induced AP by inhibiting NF-&kgr;B activation.
Repeated bouts of exercise alter the blood lactate-RPE relation.

PURPOSE To examine the effects of repeated bouts of exercise on the blood lactate [HLa]-ratings of perceived exertion (RPE) relation. METHODS Six moderately trained males were studied on two occasions: a sequential exercise bouts day (SEB: 1000 h, 1130 h, and 1300 h) and a delayed exercise bouts day (DEB: 1000 h, 1400 h, and 1800 h). Each of the three exercise bouts within a given condition were 30 min in duration at the power output (PO) associated with 70% of VO2peak on a cycle ergometer. A standardized meal was provided at 0600 h. VO2, PO, HR, and RER were recorded every min during exercise and blood [HLa] and RPE were measured every 5 min during exercise. RESULTS A 2 x 3 analysis of variance with repeated measures revealed that blood [HLa] decreased significantly with each repeated exercise bout (X +/- SEM: bout 1: SEB = 3.5 (0.3), DEB = 3.8 (0.4); bout 2: SEB = 2.6 (0.3), DEB = 2.8 (0.3); bout 3: SEB = 2.0 (0.2), DEB = 2.1 (0.4); mM). No differences were observed in the blood [HLa] response to repeated bouts of exercise between SEB and DEB. RPE-peripheral (legs, RPE-L) was higher during bout 3 compared with bout 1 (P <0.05) (bout 1: SEB = 11.8 (0.8), DEB = 12.3 (0.2); bout 2: SEB = 12.3 (0.5), DEB = 13.3 (0.4); bout 3: SEB = 13.5 (0.8), DEB = 14.0 (0.7); RPE-central (chest and breathing, RPE-C) was not affected by repeated bouts of exercise, whereas RPE-Overall (RPE-O) was higher during bout 3 compared with bouts 1 and 2 (P < 0.05) (bout 1: SEB = 12.5 (0.2), DEB = 12.3 (0.4); bout 2: SEB = 12.8 (0.4), DEB = 12.7 (0.4); bout 3: SEB = 13.7 (0.7), DEB = 13.2 (0.3)). No interaction for RPE x condition was observed. HR increased with repeated bouts of exercise with HR during exercise bout 3 being higher than HR during exercise bout 1 (164 vs. 156 bpm, P < 0.05). There was also a strong trend for HR during exercise bout 3 to be higher than HR during exercise bout 2 (P < 0.06). A trend for a reduction in VO2 with repeated exercise was observed (P < 0.07), with the reduction apparently related to the SEB condition (P < 0.12 for VO2 x condition). PO and kcal.min-1 were not affected by repeated bouts of exercise. RER decreased significantly with each repeated bout of exercise (from RER = 0.96 to RER = 0.89, P < 0.05) with no difference observed between SEB and DEB. CONCLUSIONS We conclude that the blood [HLa]-RPE relation is altered by repeated bouts of exercise and that this alteration does not appear to be affected by recovery time between exercise bouts (up to 3.5 h of recovery). These data suggest that, after the first exercise bout, RPE should not be used to produce a specific blood [HLa] on subsequent exercise bouts.
Beta Is Dead: Evidence from China

Countless empirical research has been conducted to test the validity of the Capital Asset Pricing Model (CAPM). There are claims that beta has no explanatory power over average stock return, thus casting doubt over the CAPM. This paper, moreover, provides evidence from China's stock market that beta fails to capture the cross-sectional variation in average stock return from September 1994 to October 1998 and thus concludes that the CAPM does not work in China.
Local accumulation of titanium released from a titanium implant in the absence of wear.

Titanium and its alloys, like the majority of metallic implant materials, release passive metal dissolution products. This raises the issues of amount and fate, i.e. transport, storage, and/or excretion of these metal dissolution products. In this paper we document titanium levels in tissues local to a commercially pure titanium implant in the absence of wear; compare these values to control tissues; and determine the relative contribution of the local accumulation to total release. Titanium fiber felts were implanted into the tibia of rabbits for periods up to 1 year. Bone and muscle tissue samples near the implant were collected. Using electrothermal atomic absorption spectrophotometry the samples were analyzed for titanium content. Compared to controls, titanium levels in the bone near the implant were elevated at 1-, 4-, and 12-month postoperative time points. The 12-month time point had higher periprosthetic bone titanium levels than both the 1- and the 4-month implant groups. Titanium levels in muscle tissue near the felt also indicated release was occurring. The data support the hypothesis that metal species released from titanium implants in the absence of wear have a limited solubility. As a result, they tend to remain in an area local to the implant.
ATRA and KL promote differentiation toward the meiotic program of male germ cells.

While it is known that Retinoic Acid (RA) induces meiosis in mouse female fetal gonads, the mechanisms which regulate this process during spermatogenesis are poorly understood. We show that the All trans RA derivative (ATRA) and Kit Ligand (KL) increase meiotic entry of postnatal mouse spermatogonia in vitro without synergism. Competence to enter meiosis is reached by spermatogonia only at the stage in which they undergo Kit-dependent divisions. Besides increasing Kit expression in spermatogonia, ATRA also upregulates KL expression in Sertoli cells. Both ATRA and KL increase the expression of Stimulated by Retinoic Acid Gene 8 and Dmc1, an early meiotic marker. A specific Kit tyrosine kinase inhibitor prevents the increase in the number of meiotic cells induced by both the two factors, suggesting that they converge on common Kit-dependent signalling pathways. Meiotic entry induced by ATRA and KL is independent from their ability to affect germ cell viability, and is mediated by the activation of PI3K and MAPK pathways through Kit autophosphorylation. ATRA-induced phosphorylation of the two downstream kinases is mediated by a non-genomic mechanism. These data suggest that RA may control the timing of meiosis by influencing both the somatic and the germ cell compartment of the postnatal testis through the activation of the KL/Kit system.