Search

Poncirus trifoliata (P. trifoliata) fruits exert phytotherapeutic effects, depending on their maturity level. However, the mechanism by which these phytotherapeutic effects are exerted remains undefined - especially in cancers. Therefore, in this study, we investigated the effects of the immature fruit extract of P. trifoliata on a B16 melanoma cell line.The effect of immature P. trifoliata extract on B16 cells was evaluated by MTT assay, cell proliferation, FACScan analysis of cell cycles, confocal imaging analysis, nuclear (Hoechst) staining, apoptosis assay (Annexin V-fluorescein isothiocyanate/propidium iodide staining), and Western blot assay. The capacity of immature P. trifoliata extract to inhibit the invasion and migration of B16 cells was assessed using the scratch-wound assay and Matrigel migration assay. The effect of immature P. trifoliata extract on mitochondrial function was determined via the mitochondrial membrane potential assay, activity, and fraction and cytosol proteins.Treating B16 cells with a methanol extract of immature P. trifoliata (MEPT) significantly inhibited cell viability, migration, and invasiveness in a dose- (p0.01) and time (p0.01)- dependent manner. MEPT arrested the cells in the G1 phase of the cell cycle and led to the activation of the PI3K/AKT/p21 pathway. Furthermore, MEPT dose-dependently induced apoptosis in B16 cells by increasing the expression of the pro-apoptotic proteins Bax and Apaf-1, while decreasing the expression of the anti-apoptotic protein, Bcl-2. MEPT treatment also decreased mitochondrial membrane potential.Immature P. trifoliata extract inhibited the growth of melanoma cells by inducing cell apoptosis through mitochondrial pathways. Therefore, further research into immature P. trifoliata extract as a potential therapeutic compound for melanoma treatment is warranted.

A simple, rapid, and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of zinc pyrithione (ZnPT) and pyrithione (PT) in shampoos. The method consisted of a liquid–liquid extraction for sample preparation. The mass spectrometer was operated in multiple reaction monitoring (MRM) mode via the positive electrospray ionization interface. A linear regression (weighted 1/x) was used to fit calibration curves over the concentration range of 50–2000 ng/mL for both ZnPT and PT. Excellent linearity (r 2 ≥ 0.9996) was achieved for all. The method was validated and found to be accurate (95.9–108.2% for ZnPT and 94.9–110.4% for PT), precise, and selective. Analytes in shampoos were found to be stable in the autosampler (6 °C for 6 h), in room temperature (for 6 h), and after three freeze–thaw cycles, and recovery of analytes was reproducible (90.8–94.6% for ZnPT and 90.2–96.3% for PT).

Summary Objectives Primary cilium is required for mechano-biological signal transduction in chondrocytes, and its interaction with extracellular matrix is critical for cartilage homeostasis. However, the role of cilia-associated proteins that affect the function of cilia remains to be elucidated. Here, we show that Dicam has a novel function as a modulator of primary cilia-mediated Indian hedgehog (Ihh) signaling in chondrocytes. Methods Cartilage-specific Dicam transgenic mouse was constructed and the phenotype of growth plates at embryonic day 15.5 and 18.5 was analyzed. Primary chondrocytes and tibiae isolated from embryonic day 15.5 mice were used in vitro study. Results Dicam was mainly expressed in resting and proliferating chondrocytes of the growth plate and was increased by PTHrP and BMP2 in primary chondrocytes. Cartilage-specific Dicam gain-of-function demonstrated increased length of growth plate in long bones. Dicam enhanced both proliferation and maturation of growth plate chondrocytes in vivo and in vitro, and it was accompanied by enhanced Ihh and PTHrP signaling. Dicam was localized to primary cilia of chondrocytes, and increased the number of primary cilia and their assembly molecule, IFT88/Polaris as well. Dicam successfully rescued the knock-down phenotype of IFT88/Polaris and it was accompanied by increased number of cilia in tibia organ culture. Conclusion These findings suggest that Dicam positively regulates primary cilia and Ihh signaling resulting in elongation of long bone.

Background Berberine (Ber), used widely as an antibacterial, antifungal, and anti-inflammatory drug, has long been used as a gastrointestinal remedy in Chinese traditional medicine. Recent reports have suggested that Ber suppresses Th17 responses that was mediated by direct actions on T cells and thymic stromal lymphopoietin production in primary mast cells. It has been suggested that Ber may be useful in treating allergic response. The purpose of this study was to assess the effects of Ber treatment on allergic inflammation in an allergic rhinitis mouse model and to examine the underlying mechanism(s). Methods BALB/c mice were divided into control, Derf with no treated (Derf), Ber treated, and Ber with anti-C25 monoclonal antibody treated (Ber + anti-CD25) groups. All mice, with the exception of the control group, were sensitized with an intraperitoneal i.p. injection of Dermatophagoides farinae (Derf). Mice in the Ber and Ber + anti-CD25 group were treated intranasally with 10 #181;g/mL. Then, 1 week after sensitization, all mice were challenged intranasally with 20 #181;g Derf for 5 consecutive days. Mice in the anti-CD25 group were treated intraperitoneally with 250 #181;g anti-CD25 monoclonal antibody 1 day before the first intra-nasal challenge with Derf. Allergic symptom scores, eosinophil counts, and serum Derf-specific IgE levels were measured. T-bet, GATA-3, interferon-g (IFN-γ), interleukin (IL)-10, IL-13, and Foxp3 expression was examined by real-time polymerase chain reaction and Western blotting. CD4⁺ CD25⁺ Foxp3⁺ T cells were assessed by flow cytometry. Results Symptom scores, serum Derf-specific IgE levels, GATA-3 mRNA levels, T-bet mRNA levels, and tissue eosinophil counts were decreased in the Ber versus the Derf group. In the Ber + anti-CD25 group, serum IL-10 levels were decreased versus the control, Derf, and Ber groups. In the Ber + anti-CD25 mAb groups, Foxp3 mRNA levels were decreased versus the control group. In the Ber group, Foxp3 mRNA levels were increased versus the control group. In the Ber group, the percentage of CD4⁺ CD25⁺ Foxp3⁺ T cells was increased versus the Derf group. The percentage of CD4⁺ CD25⁺ Foxp3+ T cells was increased in the Ber versus the Derf groups. Conclusions In our study, Ber reduced allergic inflammation significantly. Moreover, our findings suggest that the mechanism of action of Ber may be via CD4⁺ CD25⁺ Foxp3⁺ Treg cells, possibly through not only by increasing their numbers but also altering their function.

Background Fracture is one of the most common and important comorbidities in rheumatoid arthritis (RA) patients, especially patients who use glucocorticoids (GC). However, bone mineral density (BMD) by dual-energy x-ray absorptiometry (DXA) which is the gold standard of diagnosing and monitoring osteoporosis is not a useful tool for predicting new fracture in RA patients. Previous studies suggested the possibility of trabecular bone score (TBS) as a useful predictor for incident fracture. Objectives We aimed to evaluate the accuracy of TBS combined with clinical risk factors or BMD for prediction of new fracture in patients with RA. Methods A total of 100 female RA patients were enrolled with assessment of TBS, BMD, and clinical risk factors for fracture. During follow-up period, we calculated the incident rate of all fractures. After dividing the patients according to the use of GCs, we compared baseline characteristics and fracture-free survival between two groups. We compared accuracies of TBS, BMD, clinical risk factors for fracture and their combinations for predicting new fractures using areas under the receiver operator characteristic (ROC) curve (AUC). Results A total of 14 fractures in 12 patients were occurred among 100 patients during follow-up (428.8 person-years): 9 among the 44 in GC users and 5 in 56 GC non-users. Incidence of fracture was not different between two groups (log-rank test, p=0.27). AUC for incident fracture prediction of TBS alone [AUC 0.54, 95% confidence interval (CI) 0.35–0.72] was comparable with TBS combined with L-spine BMD (AUC 0.54, 95% CI 0.36–0.71) or with hip BMD (AUC 0.55, 95% CI 0.37–0.73). Accuracy for prediction of new fracture is increased when TBS was combined with age and history of previous fracture (AUC 0.74, 95% CI 0.62–0.85). In GC users, history of previous fracture alone (AUC 0.79, 95% CI 0.62–0.97) showed the best accuracy for predicting new fracture among TBS, BMD, clinical risk factors for fracture and their combinations. Conclusions TBS combined with age and previous history of fracture showed the highest accuracy for predicting new fracture compared to TBS or BMD alone or their combinations in RA patients. In GC users, history of previous fracture alone showed the highest accuracy for predicting new fracture. References Briot K, Paternotte S, Kolta S, Eastell R, Reid DM, Felsenberg D, et al. Added value of trabecular bone score to bone mineral density for prediction of osteoporotic fractures in postmenopausal women: the OPUS study. Bone 2013;57:232–6. McCloskey EV, Oden A, Harvey NC, Leslie WD, Hans D, Johansson H, et al. A Meta-Analysis of Trabecular Bone Score in Fracture Risk Prediction and Its Relationship to FRAX. J Bone Miner Res 2016;31:940–8. Disclosure of Interest None declared

Exposures to metal toxicants in the environment disrupt normal physiological functions and have been linked to the development of a myriad of human diseases. While the molecular and cellular mechanisms underlying metal toxicities remain to be fully understood, it is well appreciated that metal toxicants induce cellular stresses and that how cells respond to the stresses plays an important role in metal toxicity. In this review, we focus on how metal exposures induce stresses in the endoplasmic reticulum (ER) to elicit the unfolded protein response (UPR). We document the emerging evidence that induction of ER stress and UPR in the development of human diseases is associated with metal exposures. We also discuss the role of the interplay between ER stress and oxidative stress in metal toxicity. Finally, we review recent advances in functional genomics approaches and discuss how applications of these new tools could help elucidate the molecular mechanisms underlying cellular stresses induced by environmental metal toxicants.

Searches for a light sterile neutrino have been performed independently by the MINOS and the Daya Bay experiments using the muon (anti) neutrino and electron antineutrino disappearance channels, respectively. In this Letter, results from both experiments are combined with those from the Bugey-3 reactor neutrino experiment to constrain oscillations into light sterile neutrinos. The three experiments are sensitive to complementary regions of parameter space, enabling the combined analysis to probe regions allowed by the Liquid Scintillator Neutrino Detector (LSND) and MiniBooNE experiments in a minimally extended four-neutrino flavor framework. Stringent limits on sin(2) 2 theta(mu e) are set over 6 orders of magnitude in the sterile mass-squared splitting Delta m(41)(2). The sterile-neutrino mixing phase space allowed by the LSND and MiniBooNE experiments is excluded for Delta m(41)(2) < 0.8 eV(2) at 95% CLs.

Background Carbon monoxide (CO) and nitric oxide (NO) each have unique roles for various inflammatory states, including inflammatory bone resorption. Although it is known that NO can induce the expression of the cytoprotective enzyme HO-1, there is no information as to whether the protective effect of CO requires NO production or whether CO must induce the expression of HO-1 to exert its functional effects. Methods Murine osteoblast cells, MC3T3E1 osteoblasts, were cultured for CO and NO-associated HO-1 experiments and were transfected with pcDNA 3, pcDNA 3-HO-1, control siRNA or HO-1 siRNA using Nucleofector. For cell death measurement, MTT and annexin V assays were used. We performed Western blotting to check the expressions of HO-1 and iNOs and measured the HO-1 enzyme activity. We also measured the amounts of nitrite and nitrate using Griess reagents. Results The increased expression of HO-1 is required for the protective effect of NO and a single treatment of CO can increase the expression of HO-1, and this is also important for the protective effect of CO in MC3T3E1 osteoblasts. CO as well as NO attenuates the TNF-α-induced apoptosis in osteoblasts. The anti-apoptotic effect of CO or NO is not mediated by cGMP, and CO has no effect on the release of NO. The inhibition of HO-1 with using the HO-1 inhibitor ZnPP or HO-1 siRNA resulted in a striking increase of apoptosis in the CO/TNF-α-treated cells. Furthermore, HO-1 overexpression showed resistance against the TNF-α-induced cytotoxicity in the MC3T3E1 osteoblasts. Conclusions There is a need for HO-1 expression to mediate the protection provided by exogenous CO or NO in osteoblasts.

In China, Japan, and Korea, placenta hominis extracts (PHEs) are used clinically for the treatment of osteoporosis. The anti-osteoporotic effect of PHEs was studied. The trabecular bone area and thickness in OVX rats decreased by 50% from those in sham-operated rats; these decreases were completely inhibited by administration of PHEs for 7 weeks. Osteoclast numbers and the osteoblast surface were enhanced in OVX rats, but PHEs had no effect on these phenomena. Serum phosphorus and alkaline phosphatase in OVX rats increased compared to those in sham-operated rats, but the increases were not affected by the administration of PHEs. Thyroxine (T4) level was stimulated in OVX rats. The extracts inhibited the T4 level in the OVX rats. These results strongly suggest that PHEs be effective in preventing the development of bone loss induced by OVX in rats.

Yuk-Hap-Tang (YHT) induces cell death in human cervical carcinoma HeLa cells. Caspase-3, -6 and -9 were markedly activated in HeLa cells treated with YHT. The preferred substrate for caspase-3 cysteine protease, PARP, was cleaved to its 85-kDa cleavage product. YHT increased the amount of the anti-apoptotic protein, Bcl-2, and the pro-apoptotic protein, Bax. Although p53 has been reported to accumulate in cancer cells in response to anticancer agents, the p53 expression level was not changed in HeLa cells treated with YHT. Manganese (Mn)-TBAP, a mitochondria-specific SOD mimetic agent and NAC/GSH (N-acetyl cysteine/reduced glutathione) reduced the YHT-induced cytotoxicity and decreased the number of the YHT-induced apoptotic cells. Furthermore, YHT reduced the expression of Mn-SOD protein and its activity in HeLa cells. The data demonstrate that YHT induces the apoptosis of human cervical carcinoma HeLa cells by intervening Mn-SOD.

β-Wollastonite glass-ceramics was prepared by using automobile waste glass and waste shell. We solved an environmental problem using automobile waste glass and waste shell. Two different powder mixtures, automobile waste glass and waste shell, were mechanically ground in a disk-type ball mill for 4 h. After milling, the mixtures were pressed into a disk 10 mm in diameter without using binder and then heated to 850°C, 950°C, and 1050°C at a rate of 5°C/min for 1 h, respectively. Crystallinity, morphological properties, and chemical composition were observed by X-ray diffraction (XRD), field emission-scanning electron microscopy (FE-SEM), and dispersive X-ray spectrometer (EDX). β-Wollastonite and sodium calcium silicate were investigated by XRD. From the results of analysis, at 1050°C for 1 h, β-wollastonite crystals were significantly observed, so we knew that formation of the highest crystallized β-wollastonite crystal is dependent on suitable heat-treated temperature. Mechanical property investigations were carried out on β-wollastonite glass-ceramics.

Summary Objective We investigated the roles of CXC chemokine ligand 12a (CXCL12a), also known as stromal cell-derived factor-1α (SDF-1α), in endochondral bone growth, which can give us important clues to understand the role of CXCL12a in osteoarthritis (OA). Methods Primary chondrocytes and tibial explants from embryonic 15.5 day-old mice were cultured with recombinant mouse CXCL12a. To assess the role of CXCL12a in chondrogenic differentiation, we conducted mesenchymal cell micromass culture. Results In tibia organ cultures, CXCL12a increased total bone length in a dose-dependent manner through proportional effects on cartilage and bone. In accordance with increased length, CXCL12a increased the protein level of proliferation markers, such as cyclin D1 and proliferating cell nuclear antigen (PCNA), in primary chondrocytes as well as in tibia organ culture. In addition, CXCL12a increased the expression of Runx2, Col10 and MMP13 in primary chondrocytes and tibia organ culture system, implying a role of CXCL12a in chondrocyte maturation. Micromass cultures of limb-bud mesenchymal progenitor cells (MPCs) revealed that CXCL12a has a limited effect on early chondrogenesis, but significantly promoted maturation of chondrocytes. CXCL12a induced the phosphorylation of p38 and Erk1/2 MAP kinases and IκB. The increased expression of cyclin D1 by CXCL12a was significantly attenuated by inhibitors of MEK1 and NF-κB. On the other hand, p38 and Erk1/2 MAP kinase and NF-κB signaling were associated with CXCL12a-induced expression of Runx2 and MMP13, the marker of chondrocyte maturation. Conclusion CXCL12a promoted the proliferation and maturation of chondrocytes, which strongly suggest that CXCL12a may have a negative effect on articular cartilage and contribute to OA progression.

Epitaxial 3C–SiC films were grown by chemical vapor deposition on the silicon-on-insulator (SOI) substrates with 20–75-nm-thick Si top layers. A relatively low growth temperature of 1150 °C and a reduced hydrogen flow rate of 1 lpm during the precarbonization process was necessary to preserve the SOI structure and thereby obtain high-quality SiC films. The transmission electron microscopy observation of the SiC/SOI structures revealed high density of misfit dislocations in the SiC film, but no dislocation within the top Si layer. The x-ray-diffraction results did not show any significant shift of the (400) SiC peak position among the SiC/Si and the SiC/SOI samples. This strongly suggests that the Si top layer is not deformed during the SiC/SOI growth and the strain within the 3C–SiC layer is not critically affected by substituting the Si substrate with the SOI substrate, even when the Si top layer is as thin as 20 nm.

Although the recent swine flu pandemic posed an international health threat, the disease was the subject of many humorous cartoons and posters. Such a response is not new. It has long been recognized that in times of such adversity humour can play a significant role in aiding communities to cope with the fear of disease and death.1 In the art associated with the cholera pandemics of the 19th century, for example, humour and fear combined with particular resonance. The cholera pandemic of 1831–1832 and its associated high mortality (31,000 deaths in Britain alone)2 may appear an inappropriate topic for cartoons but its arrival in Sunderland in October 1831 provided an opportunity for satirists to excel. The cholera cartoons highlighted the disagreement between doctors on the diagnosis and treatment of cholera, public mistrust of the medical profession and their ineffective treatment, and political attacks on government policies. Several of these themes were included in Robert Cruikshank's Random Shots (No 1) A Cholera Doctor ( Figure 1) and Robert Cruikshank's Random Shots (No 2) A Cholera Patient ( Figure 2). These humorous prints illustrate contrasting experiences during the pandemic – the affluent medical profession thrives while the destitute public withers. Robert Cruikshank (1789–1856) was born in London and after a brief commission in the East India Company he returned to the capital where he became a book illustrator and cartoonist. Although not as successful an artist as his father Isaac or his brother George, with whom he often collaborated, he was equally adept at ridiculing topical events. Figure 1 Robert Cruikshank's Random Shots (No 1) A Cholera Doctor, c1832 (coloured etching). Reproduced with permission from Yale University, Harvey Cushing/John Hay Whitney Medical Library (in colour online) Figure 2 Robert Cruikshank's Random Shots (No 2) A Cholera Doctor, c1832 (coloured etching). Reproduced with permission from Yale University, Harvey Cushing/John Hay Whitney Medical Library (in colour online) In Robert Cruikshank's Random Shots (No 1) A Cholera Doctor the artist exaggerates the public's mistrust in doctors and in particular the belief that they had a pecuniary interest in prolonging the epidemic. On 14 February 1832, as cholera was making its first appearance in London, an anonymous letter writer (‘Theta’) to The Times accused the Central Board of Health, which had been established in June 1831 to investigate the causes of cholera and organize preventive measures, of paying its medical officers a fee of 20 guineas a day while the epidemic lasted and criticized ‘the profit derived from cholera-phobia by the profession at large’.3 Following a week of heated debate on the subject, an editorial in The Times on 21 February 1832 supported the criticism of the medical profession and even commented that ‘it is doubtful whether any really contagious disease exists’.4 In Cruikshank's print the wealthy doctor helping himself to a rather generous portion of pie etched with the words ‘£20 per day’ (perhaps an added insult to doctors since professionals were paid in guineas) is also fixated by the cascade of coins. ‘Lancet’ is engraved on the sabre-shaped knife, alluding to the journal's defence of the Central Board of Health following The Times' letter. In a leading article, The Lancet criticized The Times for its ‘slanderous attacks’ on the Board of Health, and the ‘atrocious falsehoods regarding the pay and emoluments of those engaged in the execution in the sanitary duties’ and revealed that six medical inspectors received payment of ‘the full sum of seven shillings and six pence per day, with 10s. 6d. a week for lodging expenses!!’5 The name of Dr Robert Daun, a Government Inspector who had been sent to the initial outbreak in Sunderland in 1831, appears on the label of the brandy bottle (‘cholera brandy’ was one of the many unproven cures or preventive measures against the disease). The Times letter had singled out Dr Daun as a beneficiary of the Board of Health's plans. In Robert Cruikshank's Random Shots (No 2) A Cholera Patient the artist turns his attention to the impoverished public and extends his ridicule of the medical management of cholera. The terrified and emaciated patient is perched next to a table propped up with human bones and laden with the standard treatment. His elbow showing through his threadbare clothes rests on a box of Blue pills (a purgative containing mercury), one of which he is holding in his hand, next to an anthropomorphic-shaped emetic bottle from which claw-like hands extend. As starvation was not a recommended treatment the ‘starvation stool’ may refer to the hardship endured by most patients and to the Day of Fasting and Humiliation held on 21 March 1832, arranged as an act of penance and resented by the poor.6 The bald-headed ‘Fee Fo Fum’ creature under the table alludes to the impending doom of the patient – ‘I smell the blood of an Englishman’. The cholera pandemic of 1831–1832 and subsequent outbreaks also caused widespread devastation on mainland Europe. The Continent's response was similar to Britain's ‘cholera-phobia’ but with an additional concern – the risk of premature burial. Reports of errors in the confirmation of death arose from the death-like appearances of severe cases and were exacerbated by the requirement for an early and quick burial.7,8 One of the most melodramatic images on this theme is L'Inhumation Precipitee [The Premature Burial] ( Figure 3) by the artist Antoine-Joseph Wiertz (1806–1865). Born in Dinant, Belgium, Wiertz studied at Antwerp Academy and moved to Paris in November 1828. In 1837, after a three-year residence in Rome, he returned to Belgium, settling first in Liege and then in Brussels in 1845. Figure 3 Antoine-Joseph Wiertz: L'Inhumation Precipitee [The Premature Burial], 1854 (oil on canvas) Musee Wiertz Museum, Brussels (in colour online) Although a fine portraitist, Wiertz became celebrated for his bizarre and often disturbing fantasy compositions in which death is a common theme. Even at the age of 15 he exhibited eccentric behaviour living as a recluse in a miserable attic room accompanied by a skeleton and a ‘cleverly painted death’s head'.9 The Premature Burial is set in a dark stone cellar or crypt the floor of which is littered with coffins and skeletal remains. In the foreground the shrouded figure of a cholera victim cries out in fear as he prises open the lid of the coffin in which he has been laid and attempts to escape, his arm desperately stretched out towards us. And yet at the centre of this nightmarish vision lies an unexpected touch of macabre humour for Wiertz has added an inscription on the side of the coffin: ‘Mort du cholera – certifie par nous docteurs [signe] sandoutes’ (death from cholera – certified by our doctors [signed] without doubt). Blending fear and humour, the painting powerfully illustrates ‘without doubt’ that the unfortunate victim has indeed been buried alive. Edgar Allan Poe's short story The Premature Burial (1844) has been cited as the stimulus for Wiertz's painting but a much more personal experience may have shaped the image. As his studies in Rome drew to a close in 1837, news reached him of a cholera epidemic in Naples. By promising the medical authorities of Rome that he would not return there, Wiertz was given permission to visit Naples where he discovered ‘the sick, but not yet dead, people being hastily buried, so great was the population’s (and Doctors') fear for the contagion' (personal communication, Brita Velghe, Musee Wiertz Museum). Even before his visit to Naples, Wiertz would have been aware of the concern about the risk of premature burial: it was a theme found in numerous continental folklore tales. In addition, during the late 18th and early 19th centuries there was considerable debate over the signs of confirmation of death resulting in several articles in the popular press. This led to preventive measures including extending the time between death and burial to 48 hours in Belgium and Austria.10 In 1790, Dr Christoph Wilhelm Hufeland, practising in Weimar, proposed a house for the dead which he called the Vitae Dubiae Asylum (the Asylum of Doubtful Life), subsequently known as a waiting mortuary.10 His asylum was designed to provide ‘the supervision of alleged corpses until they could safely be declared dead’.10 The first one opened in Weimar in 1792, followed by many more throughout Germany and Austria during the next 90 years. A 14 ‘corpse bedded’ waiting mortuary in Brussels was in use until the 1870s.10 Wiertz, who died from septicaemia on 18 June 1865, had arranged with the Belgian government an exchange of his paintings for the construction of a large studio (now the Musee Wiertz Museum, Brussels) built to house his art. In 1868 the museum, with its images of disease and death including The Premature Burial, became part of the Royal Museums of Fine Arts of Belgium. Cruikshank and Wiertz are only two of the many artists whose work offers a window into the impact of such catastrophic diseases. As future pandemics appear, even in this era of multimedia, there remains an important role for art in documenting not only the physical suffering but also the use of humour as a coping mechanism to alleviate fear of disease and death.

s / Osteoarthritis and Cartilage 21 (2013) S63–S312 S129 (AA)) to investigate the mineralization phase of late chondrogenesis. Monolayers of MC3T3-E1 were cultured for 21D in aMEM + 10%FBS, supplemented with BGP and AA. mRNA expression of typical chondrogenesis markers (Aggrecan (Agg), type II (Col2a1) and X (Col10a1)) collagens and osteogenic markers (Osterix (Osx), Osteocalcin (Ocn) and Osteopontin (Opn)) were assessed by quantitative RT-PCR. We also assessed mRNA expression of Wnts (-4, -5a, -5b, -11) and BMPs (-2, -4, -6, -7). Quantification of Alcian Blue, Safranin O, Sirius red and Alizarin red staining were used to evaluate proteoglycans, collagens and mineralized content respectively. Alkaline phosphatase (ALP) activity allowed to monitor osteogenesis. SDS-PAGE was used to assess the global glycosylation status. Western blot allowed to check canonical (bcatenin) and non-canonical (Calmodulin-Kinase II) Wnt signaling, as well as BMP signaling (Smad and MAPK). Results: During early proliferation (D1-7) and late proliferation/prehypertrophy phases (D7-14) of chondrogenesis, Col2a1 and Agg mRNA strongly increased, but to a lesser extent in COG5-. Proteoglycan staining intensity was lower in COG5-. BMP-4 levels progressively increased and were significantly higher in COG5-. This was consistent with increased phosphorylation of Smad 1/5/8. Wnt-4, -5a and -5b mRNA were evenly increased in both controls and COG5-. BMP-6 and Col10a1 mRNA were increased at D14, but were significantly lower in COG5-. Wnt-11 mRNA increased in controls, but not in COG5-. In mineralization phase (D14 to D21), Col10a1 mRNA and mineralization were strongly increased, but to a lesser extent in COG5-. During osteogenesis, Opn, Osx and Ocn mRNA were significantly higher in controls, alike Alizarin red staining and ALP activity. Protein analysis suggested differences in global glycosylation status, in particular of Wnt ligands. Conclusions: COG5 defect reduces chondrogenesis and osteogenesis. COG5 particularly affected glycosylation andWnt signaling, underlining its critical role in the fate of cells from the articular chondrocyte-bone unit in the course of OA. 232 ROLE OF INTERLEUKIN-10 IN ENDOCHONDRAL BONE FORMATION G. Kim y,z, Y.-K. Jung z, H.-R. Park z, E.-J. Lee z, J.-Y. Choi x, F. Beier {,k, S.-W. Han y,z. yDept. of Rheumatology, Daegu Fatima Hosp., Daegu, Republic of Korea; z Lab. for Arthritis and Bone Biology, Fatima Research Inst., Daegu, Republic of Korea; xDept.s of Biochemistry and Cell Biology, Sch. of Med., WCU program, Kyungpook Natl. Univ., Daegu, Republic of Korea; kDept. of Physiology and Pharmacology, Univ. of Western Ontario, London, ON, Canada; {Children's Hlth.Res. Inst., London, ON

BACKGROUND & AIMS: The mechanism(s) by which sucralfate heals duodenal ulcers remains unclear. The aim of this study was to determine the effect of sucralfate on Helicobacter pylori infection and on the accompanying hypersecretion of gastric acid the infection induces in patients with duodenal ulcer. METHODS: Basal and gastrin-releasing peptide (GRP) stimulated gastrin release and acid secretion. H. pylori density, gastric urease activity, and severity of gastritis were studied in patients with duodenal ulcer who were positive for H. pylori before, during, and after 4 weeks' treatment with sucralfate (2 g twice daily). RESULTS: The density of H. pylori decreased by 70% during sucralfate treatment and returned to the pretreatment level after discontinuation of therapy. This suppression of H. pylori infection was accompanied by an 80% decrease in gastric urease activity. GRP- stimulated plasma gastrin concentrations, GRP-stimulated acid output, and basal acid output all decreased by approximately 50% during sucralfate therapy and returned to pretreatment levels after treatment was discontinued. CONCLUSIONS: These findings indicate that sucralfate markedly suppresses H. pylori infection and the accompanying hypersecretion of acid the infection induces in patients with duodenal ulcer. These effects are likely to be important mechanisms by which the drug promotes duodenal ulcer healing. (Gastroenterology 1996 Mar;110(3):717-24)

1. 65Cu/63Cu stable-isotope ratios have been measured in blood serum after oral administration of the stable isotope 65Cu. The incorporation of the isotope into the plasma protein pool was followed at various times for up to 3 days. The resulting patterns of enrichment in healthy control subjects, in Wilson's disease patients and in heterozygotes for the Wilson's disease gene, were similar in appearance to those found by others using copper radioactive isotopes. After an initially high enrichment at 2h after dosage, the Wilson's disease cases, in contrast to the control subjects, did not show a secondary rise in isotope enrichment of the plasma pool after 72 h, demonstrating a failure to incorporate copper into caeruloplasmin. The Wilson's disease heterozygotes had variable degrees of impairment of isotope incorporation, not always distinguished from those of control subjects. 2. The stability of the isotope also permits the copper tracer to be followed for a longer period. Ten healthy subjects were studied for over 40 days, allowing the biological half-time of an oral dose of copper to be determined (median 18.5 days, 95% confidence interval 14–26 days). Known heterozygotes for the Wilson's disease gene were found to have a significantly increased biological half-time for removal of copper from the plasma pool (median 43 days, 95% confidence interval 32–77 days). 3. The incorporation of 65Cu in patients with diseases of the liver (other than Wilson's disease) was found to be similar to that in control subjects, aiding differential diagnosis.

Streptococcus mutans, a major etiological agent of dental caries, frequently causes systemic disease, such as subacute bacterial endocarditis, if it enters the bloodstream. In this study, the production pathways of the proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), induced by S. mutans in mouse macrophage were examined using a quantitative real-time polymerase chain reaction and an enzyme-linked immunosorbent assay. The S. mutans stimulated the expression of TNF-α and IL-1β mRNA at a multiplicity of infection of 1 : 100, which increased at 2 and 4 h, respectively, to 24 h. It also induced the production of high levels of the TNF-α and IL-1β proteins, which increased at 2 h and reached a peak at 4 and 24 h, respectively. Nuclear factor-κB (NF-κB) was activated and reached a maximum level 30 min after the S. mutans treatment. The expression of TNF-α and IL-1β mRNA and protein was suppressed by the treatment with pyrrolidine dithiocarbamate, an NF-κB inhibitor. The S. mutans-induced TNF-α expression was suppressed by the presence of SB203580, a p38 mitogen-activated protein (MAP) kinase inhibitor, or SP600125, a Jun N-terminal kinase (JNK) MAP kinase inhibitor. On the other hand, IL-1β expression was inhibited by extracellular signal-regulated kinase (ERK)/p38/JNK MAP kinase inhibitor pretreatment. In addition, TNF-α production was suppressed more in the Toll-like receptor 2(-/-) (TLR2(-/-)) macrophages than in the TLR4(-/-) macrophages, whereas IL-1β production was suppressed more in the TLR4(-/-) macrophages than in the TLR2(-/-) macrophages. These results show that S. mutans stimulates the production of TNF-α and IL-1β in the mouse macrophage cell line, RAW 264.7, by activating ERK/p38/JNK, and NF-κB through TLR2 and TLR4, respectively.

John Lavery's The First Wounded, London Hospital, August 1914 records a memorable event in the First World War. This painting and the archives of the Royal London Hospital provide a fascinating insight into the nursing and medical care of these early war casualties.

Electrical switching of terahertz radiation through nano antennas on VO2 thin film is demonstrated and is compared with bare VO2. Rectangular apertures act as slot antennas which attract terahertz radiation when VO2 is in semi‐conducting state. These antennas are turned‐off when VO2 becomes metallic by bias, giving an enhanced control of transmission.

OBJECTIVE--To compare percutaneous endoscopic gastrostomy and nasogastric tube feeding in patients with persisting neurological dysphagia. DESIGN--Randomised 28 day study of inpatients requiring long term enteral nutrition. SETTING--Three Glasgow teaching hospitals. SUBJECTS--40 patients with dysphagia for at least four weeks secondary to neurological disorders: 20 patients (10 women) were randomised to nasogastric feeding and 20 (eight women) to endoscopic gastrostomy. MAIN OUTCOME MEASURES--Treatment failure (blocked or displaced tubes on three or more occasions or refusal to continue treatment); duration of feeding; intake of liquid diets; complications; nutritional status at end of trial. RESULTS--One patient in each group died before starting feeding. Treatment failure occurred in 18 of the 19 nasogastric patients and in none of the gastrostomy group. The mean (SE) duration of feeding for the nasogastric group was 5.2 (1.5) days. No complications occurred in the nasogastric group but three (16%) of the gastrostomy group developed minor problems (aspiration pneumonia (two patients) wound infection (one)). Gastrostomy patients received a significantly greater proportion of their prescribed feed (93% (2%)) compared with the nasogastric group, (55% (4%); p less than 0.001) and also gained significantly more weight after seven days of feeding (1.4 (0.5) kg v 0.6 (0.1) kg; p less than 0.05). Analyses at days 14, 21, and 28 were not possible due to the small numbers remaining in the nasogastric group. CONCLUSION--Percutaneous endoscopic gastrostomy tube feeding is a safe and effective method of providing long term enteral nutrition to patients with neurological dysphagia and offers important advantages over nasogastric tube feeding.

The prevalence and clinical features of Wilson's disease in Scotland were investigated. Thirty three cases were identified but adequate information was available on only 28. In 1989, the prevalence rate was 4 per million. Ten patients with a mean (SEM) age of 18 (1.9) years presented with neurological symptoms, 12 patients aged 14 (1.7) years presented with hepatic symptoms, and six patients aged 12 (0.9) years were asymptomatic siblings of patients with Wilson's disease. Nine (56%) of the 16 patients who underwent liver biopsy on presentation were found to have cirrhosis. Penicillamine treatment was stopped in nine patients because of: abnormal peripheral blood count (6), rash (2), and patient's own choice (1). Nineteen patients were alive in 1989 -12 were well, one had chronic liver failure, four chronic neurological disabilities, and two had both chronic liver failure and neurological disabilities. Twelve patients died from: complications of chronic liver failure (2), acute liver failure (4), pneumonia associated with immobility (4), and other causes (2). Several patients who died had received incomplete medical supervision.

Investigation using DNA markers in a family with Wilson disease revealed that an apparently normal child of 10 years of age with non-diagnostic copper biochemistry had the disease. The procedure used linked restriction fragment length polymorphic markers. Demonstration of increased liver copper concentration from a liver biopsy confirmed the diagnosis and the child was started on chelation therapy. Two other asymptomatic siblings were shown, using the same techniques, not to have the disease. Similar analysis was carried out on another family with just one index case.

We present a detailed study of the effects of ion bombardment on the optoelectronic properties of a‐Si:H films. Two series of samples were deposited from a rf glow discharge at 30 and 100 mTorr of silane pressure, corresponding to two different deposition conditions. The energy of the ions impinging on the substrate was increased by applying a negative dc bias in steps of 25 V to the substrate holder. The increase of the substrate bias from 0 to −100 V had no effect on the deposition rate of a‐Si:H at 30 mTorr, whereas a factor of 2 decrease was observed for deposition at 100 mTorr. The density of states of the a‐Si:H films, determined by photothermal deflection spectroscopy and by the constant‐photocurrent method, decreased as the substrate bias was increased up to −50 V, especially for the series deposited at 100 mTorr. At the same time the valence‐band tail became sharper. These observations are consistent with the improvement of the electron drift‐mobility deep‐trapping‐lifetime (μdτd)e product, deter...

The radiolysis of cyclohexene in the presence of CO (1−10 × 10 5 Pa pressure) leads to the formation of: 2-cyclohexene-1-one; 2-cyclohexene-1-o1; bi-2-cyclohexene-1-y1; 3-cyclohexyl-cyclohexene; cyclohexanone; cyclohexanol; cyclohexane and bicyclohexyl; in addition to traces of formaldehyde; 2- and 3-cyclohexene-1-carboxaldehyde; cyclohexane-carboxaldehyde and dicyclohexylketone. The G i -value of each individual product was found to be a function of the CO concentration in the solution, e.g. G i (2-cyclohexane-1-one) = 1.05 at 1 × 10 5 Pa CO and 10.30 at 1 × 10 6 Pa CO, respectively (1 atm = 1.013 × 10 5 Pa). Experiments with deoxygenerated cyclohexane were also carried out, and the final products were analysed for comparative purposes. Probable reaction mechanisms are presented as an explanation of the results obtained.

Tanshinone I, tanshinone IIA, cryptotanshinone, and dihydrotanshinone are compounds that have been isolated from the root of Salvia miltiorrhiza (SM), which is also known as "Danshen." The SM extract has been used successfully in China for treating postmenopausal syndrome. Furthermore, it was previously reported that SM had inhibitory effect on osteoporosis in ovariectomized rats. Another study reported that the four components, tanshinone I, tanshinone IIA, cryptotanshinone, and dihydrotanshinone, prevented osteoclast function in an in vitro system. However, there are no reports of a correlation between SM and its components on osteoporosis and osteoclast function. This study was undertaken to examine the effect of SM on osteoclastogenesis and osteoblast differentiation, which are two important markers of the bone physiology. Through a rapid, sensitive and specific isocratic liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantitative determination of four diterpenoids, tanshinone I, tanshinone IIA, cryptotanshinone, and dihydrotanshinone in SM, the authors tried to correlate the amount of tanshinone compounds in SM into the antiosteoclast activity. The SM fraction (methanol and ethanol isolated) with a low concentration of tanshinone IIA (1 mug/mL) had no effect on the alkaline phosphotase activity (osteoblast differentiation), but completely inhibited osteoclastogenesis. Although the tanshinone compound itself showed similar effects, the concentrations of commercially available tanshinone (diterpenoids, tanshinone I, tanshinone IIA, cryptotanshinone, and dihydrotanshinone) needed for antiosteoclast activity was almost 1000 times more than that of tanshinone in SM fraction. This suggests that there are other unknown compounds in the SM extract that have a synergistic effect with tanshinone. These results also suggest that tanshinone can be a good marker compound to explain the antiosteoporotic function of SM.

The minimum primary structural requirement for a tripeptide to serve as a substrate for oligosaccharyl transferase is the sequence -Asn-X-Ser/Thr-. In the present study the activities of three structurally different tripeptides containing acceptor sequences for oligosaccharyl transferase were compared in three systems: Xenopus oocytes, in which they were introduced into the cytoplasm by microinjection, cultured mammalian cells, and isolated rat liver microsomes. In the last two systems, the peptides were added exogenously to the culture or to the incubation medium, respectively. On the basis of lectin column and paper chromatographic analysis it was established that the microinjected acceptor tripeptides were glycosylated in Xenopus oocytes. However, lectin column analysis and retention of sensitivity to endoglycosidase H revealed that none of the three glycopeptides was processed to complex oligosaccharide chains and none was subsequently secreted. Rather, over a 24-h period the glycopeptides were degraded. Chloroquine was found to block this degradation process, but even under these conditions, the glycopeptides were not secreted into the medium. In the isolated microsomes the glycosylation of the acceptor tripeptides was time-dependent and the tripeptide with an iodotyrosine residue in the X position was found to be a poor substrate. When added to cultured mammalian cells, all three of the tripeptides were taken up, glycosylated, and subsequently secreted. These results are discussed in the context of the wide differences in glycosylation of the three peptides and their lack of secretion after glycosylation in Xenopus oocytes.

AIMS: To measure insulin-like growth factor-I (IGF-I) concentrations in the presence and absence of Helicobacter pylori infection and in response to eradication of the organism. METHODS: An enzyme linked immunosorbent assay was used to measure gastric and fasting serum concentrations of IGF-I in 17 patients with and 11 without H pylori infection. Repeat assessments were performed in the infected patients six weeks after they received a two week course of bismuth chelate, metronidazole, and amoxycillin. RESULTS: IGF-I was detected at very low concentrations in gastric juice and in mucosal incubates. The median serum IGF-I concentration was 88 micrograms/l in the patients infected with H pylori compared with 90 micrograms/l in the non-infected controls; IGF-I concentrations dropped to 77 micrograms/l following eradication therapy (p = 0.014). CONCLUSION: The similarity in baseline IGF-I concentrations in the presence and absence of H pylori suggests that their subsequent drop after treatment is more likely to be due to the treatment.

At Christmas many doctors experience brief but well intentioned and tangible gestures of successful patient-doctor relationships. Various patients' gifts, such as boxes of chocolates or bottles of wine, are always appreciated by the recipient. Although doctors may receive more permanent gifts, very few will have had the delight of having their portrait painted by their patient. Artists from different ages and cultures—Goya, Kahlo, Bellany, Van Gogh, Munch, and Dadd—have all had the desire to record their special relationship with their doctors. The Spanish artist Francisco Goya (1746-1828) has left the most moving testimony of his gratitude for the close relationship with his doctor in Self Portrait with Dr Arrieta (1820) (fig 1). In Seville, in the autumn of 1792, Goya developed a sudden serious illness which included dizziness, weakness, delirium, sickness, abdominal pain, deafness, and partial blindness.1 2 The name of his doctor has not been recorded. He convalesced in Cadiz, where in March 1793 his friend Sebastian Martinez reported: “Goya is slightly better but progress is sadly slow. The noises in his head and his deafness have not passed away; however, his sight has improved and he no longer has fits of dizziness and can walk up and down stairs without difficulty.” On his return to Madrid in July 1793 Goya was completely deaf. Various diagnoses of this serious illness have been offered: syphilis, lead poisoning, cerebrovascular disease, acute infection of the central nervous system, and the rare condition of Vogt-Koyanagi-Harada syndrome—temporary inflammation of the uveal tract associated with permanent deafness.2 Fig 1 Goya's Self Portrait with Dr Arrieta , 1820. Minneapolis Institute of Arts, The Ethel Morrison Derlip Fund In 1819 Goya had a second serious illness. Little information is available either on the nature of the illness or on Dr Arrieta's treatment. The painting is the main …

Mouse immune sera obtained by immunization with Fusobacterium nucleatum and then Porphyromonas gingivalis demonstrated an impaired binding capacity to P. gingivalis-biofilm and lower avidity to P. gingivalis when compared with sera obtained from mice immunized with P. gingivalis alone.

Maspin is a recently described member of the serpin family or protease inhibitors that is known to be a tumour suppressor gene product. Loss of maspin expression has been found in most breast cancer cases and is correlated with cell motility and tumour invasiveness. However, its precise role in human breast cancer remains to be discovered. We aimed to evaluate the role of maspin in early-stage breast cancer.We analysed the expression of maspin in 192 stage I and II primary breast cancers by immunohistochemistry. Of these cases, 34.4% showed maspin expression. Maspin expression was more frequently found in invasive ductal carcinoma (36.4%) than in invasive lobular carcinoma (7.1%). High maspin expression was demonstrated in breast cancers showing high histological grade or lymphocyte-rich stroma (P0.05). Maspin expression was not associated with overall and disease-free survival rate of breast cancer.The results indicate that different biological mechanisms may be responsible for maspin expression in histologically distinct types of breast cancer. Our survey suggests that maspin expression in breast cancer might have a compensatory role rather than prognostic one.

The light-induced generation of defects in a-Si:H(F) was saturated by a few hours of illumination with Kr-ion-laser light soaking (λ=647.1 nm) near room temperature. The time to reach saturation scales roughly with 1/G2, but the saturation value is essentially independent of the illumination intensity. Therefore, the saturation is not due to thermal annealing. The saturation values of the light-induced defect density in 37 a-Si:H(F) samples which had been grown in six different reactors over a range of conditions were measured. These a-Si:H(F) samples have annealed-state defect densities in the range of 1.1×1015 to 1.6×1016 cm-3, Urbach energies of 42-62 meV, and Tauc optical bandgaps of 1.61 to 1.83 eV. The saturation value rises from 5×1016 to 2×1017 cm-3 with increasing optical gap and total hydrogen content, but it is not correlated with the Urbach energy or with the annealed-state defect density.

Among fluoroquinolone antibiotics, ofloxacin (OFL) and norfloxacin (NOR) have piperazinyl groups but flumequine (FLU) does not have this substitutent. The emission spectra of OFL and NOR are strong, broad structureless bands with large Stokes' shifts in water but the emission intensities are very weak in organic solvents. Thus we find that these compounds exist as different chemical species in various solvents. A continuous red shift in the emission bands for OFL and NOR is observed as the water concentration within the aerosol-OT (AOT; sodium 1,4-bis[2-ethylhexyl]sulfosuccinate) micelle increases or temperature of this solution rises. From the fluorescence anisotropy measurements of OFL and NOR, we assume the intramolecular charge transfer after excitation from the nitrogen of the piperazinyl group to the keto oxygen. Theoretical calculations further support this observation. Multifrequency phase and modulation experiments and time-resolved emission spectra clearly show the occurrence of intramolecular charge transfer and the subsequent nanosecond water reorganization around OFL or NOR in the AOT micelle. Upon increasing the water concentration within the AOT micelle, the relaxation rate increases because of the large amount of free water. The emission spectra of FLU do not exhibit any significant response to the physical properties of their environment.

The adverse effects of exposure to environmental tobacco smoke (ETS) have been documented extensively.1 Homes are a major source of ETS exposure, particularly those with smokers. Exposure can be reduced by banning smoking inside homes.2–4 Despite having one of the highest smoking rates among men worldwide,5 little is known about home smoking policies in the Republic of Korea. This study examined the prevalence of home smoking bans in Seoul and their association with ETS exposure at home. In 2002, telephone interviews were completed with 500 Seoul residents aged 18 years or older, using random digit dialling and stratification by gender. A total of 50% of eligible respondents were interviewed. Respondents indicated whether no one is allowed to smoke in your home; only special guests are allowed to smoke; people are allowed to smoke only in certain areas; or people allowed to smoke anywhere. Respondents were classified as smokers if they had smoked 100 or more cigarettes and …

We report a study of the saturated light-induced defect density Ns,sat in 37 hydrogenated (and in part fluorinated) amorphous silicon [a-Si:H(F)] films grown in six different reactors under widely different conditions. Ns,sat was attained by exposing the films to light from a krypton ion laser (λ=647.1 nm). Ns,sat is determined by the constant photocurrent method and lies between 5×1016 and 2×1017 cm−3. Ns,sat drops with decreasing optical gap Eopt and hydrogen content cH, but is not correlated with the initial defect density Ns,ann or with the Urbach tail energy Eu. We discuss our results within the framework of existing models for light-induced defects.

Home enteral nutrition (HEN) is an established method of long term nutritional support. Many patients receiving HEN have Crohn's disease complicated by intestinal failure and malnutrition, including magnesium deficiency. It is unknown if HEN can correct magnesium deficiency or if patients on HEN can become magnesium deficient. We measured total magnesium intake in nine patients receiving HEN, and assessed their magnesium status. Two patients had magnesium intakes below the recommended dietary allowance of 15 mmol/day. Four patients (44%) had biochemical evidence of magnesium deficiency, although no patient had clinical signs of magnesium deficiency. Several magnesium deficient patients used a liquid feed which had a low magnesium content. Patients on HEN should have their magnesium status checked regularly and may require magnesium supplements.

Licochalcone A (1), ombuin (2), and 5,7-dihydroxy-3',4'-dimethoxyflavanone (3) were isolated from the aerial parts of Pogostemon cablin by cytotoxicity-guided fractionation. Compound 1 showed in vitro cytotoxicity and Pl-PLC gamma 1 inhibition activity. Treatment of promyelocytic leukemia cells (HL-60) with compound 1 induced terminal differentiation with the generation of monocyte using nonspecific acid esterase assay.

Cellular and molecular characterization of osteoclasts (OCL) has been extremely difficult since OCL are rare cells, and are difficult to isolate in large numbers. We used the tartrate-resistant acid phosphatase promoter to target the bcl-XL and/or Simian Virus 40 large T antigen (Tag) genes to cells in the OCL lineage in transgenic mice as a means of immortalizing OCL precursors. Immunocytochemical studies confirmed that we had targeted Bcl-XL and/or Tag to OCL, and transformed and mitotic OCL were readily apparent in bones from both Tag and bcl-XL/Tag mice. OCL formation in primary bone marrow cultures from bcl-XL, Tag, or bcl-XL/Tag mice was twofold greater compared with that of nontransgenic littermates. Bone marrow cells from bcl-XL/Tag mice, but not from singly transgenic bcl-XL or Tag mice, have survived in continuous culture for more than a year. These cells form high numbers of bone-resorbing OCL when cultured using standard conditions for inducing OCL formation, with approximately 50% of the mononuclear cells incorporated into OCL. The OCL that form express calcitonin receptors and contract in response to calcitonin. Studies examining the proliferative capacity and the resistance of OCL precursors from these transgenic mice to apoptosis demonstrated that the increased numbers of OCL precursors in marrow from bcl-XL/Tag mice was due to their increased survival rather than an increased proliferative capacity compared with Tag, bcl-XL, or normal mice. Histomorphometric studies of bones from bcl-XL/Tag mice also confirmed that there were increased numbers of OCL precursors (TRAP + mononuclear cells) present in vivo. These data demonstrate that by targeting both bcl-XL and Tag to cells in the OCL lineage, we have immortalized OCL precursors that form bone-resorbing OCL with an efficiency that is 300-500 times greater than that of normal marrow.