The proteasome inhibitor bortezomib may increase osteoblast-related markers in multiple myeloma (MM) patients; however, its potential osteoblastic stimulatory effect is not known. In this study, we show that bortezomib significantly induced a stimulatory effect on osteoblast markers in human mesenchymal cells without affecting the number of osteoblast progenitors in bone marrow cultures or the viability of mature osteoblasts. Consistently we found that bortezomib significantly increased the transcription factor Runx2/Cbfa1 activity in human osteoblast progenitors and osteoblasts without affecting nuclear and cytoplasmatic active ␤-catenin levels. IntroductionMultiple myeloma (MM) is a plasma cell malignancy characterized by the high capacity to induce osteolytic bone lesions. 1 Hystomorphometric studies showed that osteoblast formation and function are profoundly impaired in MM patients and critical in the bone lesion development. [1][2][3] Several mechanisms are involved in MMinduced osteoblast suppression 1,4,7 including the production of Wnt inhibitors as DKK-1 or sFRP-2 5,6 or the impairment of osteoblast formation and differentiation through the block of the critical osteoblast transcription factor Runx2/Cbfa1. 7 In turn, osteoblastic cells also regulate myeloma cell growth 8,9 and the increase of bone formation in mice results in a reduction of tumoral burden. 10 Recent data suggest that ubiquitin-proteasome pathway, which is the major cellular degradative system and therapeutic target in myeloma cells, 11 also regulates osteoblast differentiation. [12][13][14] The ubiquitin-proteasome pathway can modulate the BMP-2 expression, 12 which can induce osteoblast differentiation through the Wnt signaling 13 and regulates the proteolytic degradation of the osteoblast transcription factor Runx2/ Cbfa1. 14 Recently, Garrett et al 12 demonstrated that proteasome inhibitors as PS1 and epoximicin stimulate bone formation in neonatal murine calvarial bones and in vivo in mice. 12 A strong correlation between the capacity of these compounds to inhibit proteasomal activity in osteoblasts and their bone-forming activity was also demonstrated. 12 Preliminary observations obtained in MM patients treated with the proteasome inhibitor bortezomib show an increase of serum bone-specific alkaline phosphatase and other osteoblast related markers suggesting a potential osteoblast stimulatory effect of this drug. [15][16][17][18] Currently it is not known whether the proteasome inhibitor bortezomib may have a direct effect on osteoblast differentiation and formation in vitro in human cultures and in vivo in MM patients. Patients, materials, and methods DrugsBortezomib was purchased from Janssen-Cilag (Milan, Italy). For in vitro studies, the drug was reconstituted in DMSO at a stock concentration of 50 mM, and this stock was diluted in medium just before use, so that the concentration of DMSO never exceeded 0.1%. The proteasome inhibitors MG-132 and MG-262 were purchased from BIOMOL International (Plymouth Meeting, PA; DBA srl, M...

Some polyphenolic catabolites, generated in vivo in the colon, were able in vitro to counteract two key features of diabetic complications, i.e. protein glycation and neurodegeneration. These observations could lead to a better control of these events, which are usually correlated with hyperglycemia.

Polybrominated diphenyl ethers (PBDEs), extensively used in the past few decades as flame retardants in a variety of consumer products, have become world-wide persistent environmental pollutants. Levels in North America are usually higher than those in Europe and Asia, and body burden is 3 to 9-fold higher in infants and toddlers than in adults. The latter has raised concern for potential developmental toxicity and neurotoxicity of PBDEs. Experimental studies in animals and epidemiological observations in humans suggest that PBDEs may be developmental neurotoxicants. Pre- and/or post-natal exposure to PBDEs may cause long-lasting behavioral abnormalities, particularly in the domains of motor activity and cognition. The mechanisms underlying the developmental neurotoxic effects of PBDEs are not known, though several hypotheses have been put forward. One general mode of action relates to the ability of PBDEs to impair thyroid hormone homeostasis, thus indirectly affecting the developing brain. An alternative or additional mode of action involves a direct effect of PBDEs on nervous system cells; PBDEs can cause oxidative stress-related damage (DNA damage, mitochondrial dysfunction, apoptosis), and interfere with signal transduction (particularly calcium signaling), and with neurotransmitter systems. Important issues such as bioavailability and metabolism of PBDEs, extrapolation of results to low level of exposures, and the potential effects of interactions among PBDE congeners and between PBDEs and other contaminants also need to be taken into account.

IntroductionAngiogenesis has a critical role in the pathophysiology and progression of multiple myeloma (MM) supporting the growth and survival of MM cells. [1][2][3][4][5] The angiogenic process in MM is sustained mainly by the overexpression of proangiogenic factors directly by MM cells including VEGF, 6 angiopoietin-1 (ANG-1), 7 osteopontin (OPN), . 9 Nevertheless, the molecular mechanisms underlying the regulation of angiogenesis in MM have not been completely elucidated.The new candidate tumor-suppressor gene inhibitor of growth family member 4 (ING4) has been recently implicated in solid tumors as a repressor of tumor growth and angiogenesis through the association with NF-B. ING4 is a nuclear factor expressed in all normal tissues and markedly reduced in glioblastoma cells and head and neck squamous cell carcinoma, with levels inversely correlated with tumor grade. 10,11 Inhibition of ING4 expression strongly promotes the growth of glioma cells in vivo, whereas its overexpression leads to growth inhibition through ING4's capability to interact with p65 subunit of NF-B. 10 Interestingly, it has been also shown that tumors lacking ING4 showed increased vascularization compared with ING4-expressing tumors. 12 Moreover ING4 down-regulated the angiogenic-related molecules including IL-8 and the hypoxia inducible factor-1␣ (HIF-1 ␣) activity in hypoxic condition through the involvement of HIF prolyl hydroxylase 2 (HPH-2) 10,13 In turn, the role of hypoxia has been recently highlighted in the promotion of angiogenesis. 14 The expression of ING4 by MM cells, as well as its potential role in MM-induced angiogenesis, has never been investigated. In this study, we evaluated the expression of ING4 in malignant MM cells and the potential relationship between ING4 and the production of proangiogenic molecules by MM cells in normoxic and hypoxic conditions, as well as its relationship with the "in vitro and in vivo" angiogenesis. Submitted February 15, 2007; accepted September 4, 2007. Prepublished online as Blood First Edition paper, September 11, 2007; DOI 10.1182 DOI 10. /blood-2007 The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked ''advertisement'' in accordance with 18 USC section 1734. For personal use only. on August 29, 2018. by guest www.bloodjournal.org From Patients, materials, and methods Cells and cell culture conditionsCell lines. Human myeloma cell lines (HMCLs) XG-6, XG-1, and JJN3 were obtained from Dr Bataille (Nantes, France). U266 was obtained from the American Type Culture Collection (Rockville, MD). OPM2 and RPMI-8226 were purchased from DSM (Braunschweig, Germany). ARP-1 and H929 were generously received from Dr Shaughnessy's laboratory (Little Rock, AR).Cell cultures. HMCLs were incubated in RPMI medium at 10% FCS (Invitrogen Life Technologies, Milan, Italy) and maintained with or without IL-6 (3 ng/mL; Endogen Woburn, MA). In a series of experiments, HMCLs were incubated with the HPH-2 in...

Oropharyngeal dysphagia (OD) is a widespread clinical condition among older adults. Although it represents a risk factor for malnutrition, dehydration and aspiration pneumonia, its assessment and contribution to functional decline is often ignored. The aim of the present study was to estimate the prevalence of OD in a large population of non-institutionalized older people and to evaluate its relationship with malnutrition and physical function. 10-item Eating Assessment Tool (EAT-10) and Mini Nutritional Assessment Short Form (MNA-SF) were used to identify the risk of dysphagia and malnutrition. Short Physical Performance Battery (SPPB) and hand-grip strength were used as functional endpoints. The relationship between risk of dysphagia and functional outcomes was tested in a multivariate regression analysis adjusted for age and sex (Model 1) and for other confounders including Mini Mental State Examination (MMSE) and polypharmacy (Model 2). Mean age of 773 subjects (61.3% female) was 81.97 years. The percentage of participants at risk of dysphagia (EAT ! 3) was 30.1%, 37.8% of subjects was malnourished (MNA-SF < 8), 46.2% was at risk of malnutrition (MNA-SF:8e11). EAT-10 was significantly and negatively associated to MNA-SF (b ¼ À0.47 ± 0.06, p < 0.0001) and the strength of the relationship was attenuated but still statistically significant in the multivariate model (b ¼ À0.28 ± 0.07, p < 0.0001). A significant and negative relationship was found between EAT-10 and SPPB and hand-grip strength in Model 1 (b ¼ À0.25 ± 0.05, p < 0.0001) and Model 2 (b ¼ À0.07 ± 0.03, p < 0.0001). After categorization of risk of dysphagia in two groups (at risk and not at risk), MNA-SF, SPPB and hand-grip strength were independently associated with higher risk of dysphagia (OR ¼ 0.91, 95%CI ¼ 0.83e0.99, p ¼ 0.03; OR ¼ 0.83, 95%CI ¼ 0.77e0.89, p < 0.0001; OR ¼ 0.96, 95%CI ¼ 0.92e0.99, p ¼ 0.02, respectively). In a large group of outpatient older individuals, we observed a significant negative association between risk of dysphagia and nutritional and physical performance, suggesting that the screening of OD, possibly supported by its assessment, should be implemented in the geriatric setting to potentially prevent the functional decline.

Osteoblast impairment occurs within multiple myeloma cell infiltration into the bone marrow. Canonical Wnt signaling activation in osteoprogenitor cells is involved in osteoblast formation through the stabilization of dephosphorylated Bcatenin and its nuclear translocation. The effects of multiple myeloma cells on Wnt signaling in human mesenchymal/ osteoprogenitor cells are unclear. In 60 multiple myeloma patients checked, we found that among the Wnt inhibitors, Dickkopf-1 and secreted frizzled-related protein-3 were produced by multiple myeloma cells. However, although multiple myeloma cells or multiple myeloma bone marrow plasma affected expression of genes in the canonical Wnt signaling and inhibited B-catenin stabilization in murine osteoprogenitor cells, they failed to block the canonical Wnt pathway in human mesenchymal or osteoprogenitor cells. Consistently, Wnt3a stimulation in human osteoprogenitor cells did not blunt the inhibitory effect of multiple myeloma cells on osteoblast formation. Consequently, despite the higher Wnt antagonist bone marrow levels in osteolytic multiple myeloma patients compared with nonosteolytic ones, B-catenin immunostaining was not significantly different. Our results support the link between the production of Wnt antagonists by multiple myeloma cells and the presence of bone lesions in multiple myeloma patients but show that myeloma cells do not inhibit canonical Wnt signaling in human bone microenvironment. [Cancer Res 2007;67(16):7665-74]

The gut microbiota could influence the pathophysiology of age-related sarcopenia through multiple mechanisms implying modulation of chronic inflammation and anabolic resistance. The aim of this study was to compare the fecal microbiota composition and functionality, assessed by shotgun metagenomics sequencing, between two groups of elderly outpatients, differing only for the presence of primary sarcopenia. Five sarcopenic elderly subjects and twelve non-sarcopenic controls, classified according to lower limb function and bioimpedance-derived skeletal muscle index, provided a stool sample, which was analyzed with shotgun metagenomics approaches, to determine the overall microbiota composition, the representation of bacteria at the species level, and the prediction of bacterial genes involved in functional metabolic pathways. Sarcopenic subjects displayed different fecal microbiota compositions at the species level, with significant depletion of two species known for their metabolic capacity of producing short-chain fatty acids (SCFAs), Faecalibacterium prausnitzii and Roseburia inulinivorans, and of Alistipes shahii. Additionally, their fecal metagenome had different representation of genes belonging to 108 metabolic pathways, namely, depletion of genes involved in SCFA synthesis, carotenoid and isoflavone biotransformation, and amino acid interconversion. These results support the hypothesis of an association between microbiota and sarcopenia, indicating novel possible mediators, whose clinical relevance should be investigated in future studies.

Background: Sex-differences have been demonstrated in the acute phase of COVID-19 infection; females (F) were found to be less prone to develop a severe disease than males (M), but few studies have assessed sex-differences in Long-Covid-19 syndrome. Aim and Results:The aim of this prospective/retrospective study was to characterize the long-term consequences of this infection from a sex-perspective. For this purpose, we enrolled 223 patients (89 F and 134 M) who experienced a SARS -CoV-2 infection.In the acute phase of the illness, females reported more frequently than males: weakness, dysgeusia, anosmia, thoracic pain, palpitations, diarrhea, and myalgia without significant differences in breathlessness, cough, and sleep disturbance. After a mean follow-up time of 5 months after the acute phase, females were significantly more likely than males to report weakness, thoracic pain, palpitations, and sleep disturbance but not myalgia and cough.At the multivariate logistic regression, women were statistically significantly likely to experience persistent symptoms such as dyspnoea, fatigue, chest pain, and palpitations.On the contrary, myalgia, cough and sleep disturbance were not influenced by sex. Conclusion:We demonstrated that females were more symptomatic than males not only in the acute phase but also at follow-up. Sex was found to be an important determinant of Long-COVID syndrome because it is a significant predictor of persistent symptoms in females, such as dyspnoea, fatigue, chest pain, and palpitations.Our results suggest the need for long-term follow-up of these patients from a sexperspective in order to implement early preventive and personalized therapeutic strategies.