The Value of Surveillance Imaging of Oral Squamous Cell Carcinoma.

The evaluation of surveillance imaging of OSCC patients is a difficult task physicians have to face daily. Multiple patients experience a recurrence of this disease, which underlines the importance of regular patient monitoring programs. Our study analysed the value of surveillance imaging, such as computed tomography (CT) and nuclear magnetic resonance imaging (NMRI), as a patient monitoring programme and its effectiveness in achieving improvement in early recurrence detection. The study comprised 125 patients, out of which 56 (n = 56) showed radiological and 69 (n = 69) showed clinical and radiological conspicuous patterns in domestic follow-ups, respectively. The use of CT and NMRI showed a significant dependence on the histological result (p = 0.03). However, the different groups showed no significant dependence on the histological result (p = 0.96). The distribution of the histological biopsies, which were taken due to radiological changes, were prone to wrong positive diagnoses (false positives) in 71 percent. To conclude, imaging modalities should be chosen for each patient individually to reduce false positives, improve the early detection of recurrence, and increase the cure rate.

Evaluation of CIN2/3 Lesion Regression in GynTect® DNA Methylation-Marker-Negative Patients in a Longitudinal Study.

Cervical intraepithelial neoplasia (CIN) grade 2/3 has a high spontaneous regression rate, especially among women ≤29 years of age. To reduce overtreatment, reliable prognostic biomarkers would be helpful. The main aim of this study was to analyze the negative predictive value of the methylation marker panel GynTect® for lesion regression. In this prospective, multicenter, longitudinal observational proof-of-concept study, women aged ≤29 years with histologically confirmed CIN2 (n = 24) or CIN3 (n = 36) were closely monitored without treatment for up to 24 or 12 months, respectively. The outcome was either regression, persistence, or progression of the lesion. For each patient, a single baseline sample (V0) for cytology, hrHPV detection and methylation analysis was taken. In a primary analysis, the negative predictive value (NPV) of a GynTect®-negative test result at V0 for regression was determined. We tested the null hypothesis NPV ≤ 70% against the alternative hypothesis NPV ≥ 90%. Twelve of the eighteen GynTect®-negative CIN2 patients showed regression (NPV = 67%, 90% CI 44-85%, p = 0.53). Of the 27 GynTect®-negative CIN3 lesions, 15 regressed (NPV = 56%, 90% CI 38-72%, p = 0.92). Although the majority of GynTect®-negative lesions regressed, the postulated NPV of ≥90% was not observed. Thus, the clinical relevance for an implementation of the GynTect® assay for patients undergoing watchful waiting remains questionable. Further studies with longer observation periods should be undertaken.

Selecting the Best Candidates for Cisplatin-based Adjuvant Chemotherapy After Radical Cystectomy Among Patients with pN+ Bladder Cancer.

A trend towards greater benefit from adjuvant chemotherapy (ACT) in pN+ bladder cancer (BCa) has been observed in multiple randomized controlled trials. However, it is still unclear which patients might benefit the most from this approach. We retrospectively analyzed a multicenter cohort of 1381 patients with pTany pN1-3 cM0 R0 urothelial BCa treated with radical cystectomy (RC) with or without cisplatin-based ACT. The main endpoint was overall survival (OS) after RC. We performed 1:1 propensity score matching to adjust for baseline characteristics and conducted a classification and regression tree (CART) analysis to assess postoperative risk groups and Cox regression analyses to predict OS. Overall, 391 patients (28%) received cisplatin-based ACT. After matching, two cohorts of 281 patients with pN+ BCa were obtained. CART analysis stratified patients into three risk groups: favorable prognosis (≤pT2 and positive lymph node [PLN] count ≤2; odds ratio [OR] 0.43), intermediate prognosis (≥pT3 and PLN count ≤2; OR 0.92), and poor prognosis (pTany and PLN count ≥3; OR 1.36). Only patients with poor prognosis benefitted from ACT in terms of OS (HR 0.51; p < 0.001). We created the first algorithm that stratifies patients with pN+ BCa into prognostic classes and identified patients with pTany BCa with PLN ≥3 as the most suitable candidates for cisplatin-based ACT. PATIENT SUMMARY: We found that overall survival among patients with bladder cancer and evidence of lymph node involvement depends on cancer stage and the number of positive lymph nodes. Patients with more than three nodes affected by metastases seem to experience the greatest overall survival benefit from cisplatin-based chemotherapy after bladder removal. Our study suggests that patients with the highest risk should be prioritized for cisplatin-based chemotherapy after bladder removal.

Patient-specific effects of soluble factors from Staphylococcus aureus and Staphylococcus epidermidis biofilms on osteogenic differentiation of primary human osteoblasts.

Due to the frequency of biofilm-forming Staphylococcus aureus and Staphylococcus epidermidis in orthopedics, it is crucial to understand the interaction between the soluble factors produced by prokaryotes and their effects on eukaryotes. Our knowledge concerning the effect of soluble biofilm factors (SBF) and their virulence potential on osteogenic differentiation is limited to few studies, particularly when there is no direct contact between prokaryotic and eukaryotic cells. SBF were produced by incubating biofilm from S. aureus and S. epidermidis in osteogenic media. Osteoblasts of seven donors were included in this study. Our results demonstrate that the detrimental effects of these pathogens do not require direct contact between prokaryotic and eukaryotic cells. SBF produced by S. aureus and S. epidermidis affect the metabolic activity of osteoblasts. However, the effect of SBF derived from S. aureus seems to be more pronounced compared to that of S. epidermidis. The influence of SBF of S. aureus and S. epidermidis on gene expression of COL1A1, ALPL, BGLAP, SPP1, RUNX2 is bacteria-, patient-, concentration-, and incubation time dependent. Mineralization was monitored by staining the calcium and phosphate deposition and revealed that the SBF of S. epidermidis markedly inhibits calcium deposition; however, S. aureus shows a less inhibitory effect. Therefore, these new findings support the hypotheses that soluble biofilm factors affect the osteogenic processes substantially, particularly when there is no direct interaction between bacteria and osteoblast.

Dysregulation of proinflammatory versus anti-inflammatory human TH17 cell functionalities in the autoinflammatory Schnitzler syndrome.

BACKGROUND: TH17 cells have so far been considered to be crucial mediators of autoimmune inflammation. Two distinct types of TH17 cells have been described recently, which differed in their polarization requirement for IL-1ß and in their cytokine repertoire. Whether these distinct TH17 phenotypes translate into distinct TH17 cell functions with implications for human health or disease has not been addressed yet. OBJECTIVE: We hypothesized the existence of proinflammatory and anti-inflammatory human TH17 cell functions based on the differential expression of IL-10, which is regulated by IL-1ß. Considering the crucial role of IL-1ß in the pathogenesis of autoinflammatory syndromes, we hypothesized that IL-1ß mediates the loss of anti-inflammatory TH17 cell functionalities in patients with Schnitzler syndrome, an autoinflammatory disease. METHODS: To assess proinflammatory versus anti-inflammatory TH17 cell functions, we performed suppression assays and tested the effects of IL-1ß dependent and independent TH17 subsets on modulating proinflammatory cytokine secretion by monocytes. Patients with Schnitzler syndrome were analyzed for changes in TH17 cell functions before and during therapy with IL-1ß-blocking drugs. RESULTS: Both TH17 cell subsets differ in their ability to suppress T-cell proliferation and their ability to modulate proinflammatory cytokine production by antigen-presenting cells because of their differential IL-10 expression properties. In patients with Schnitzler syndrome, systemic overproduction of IL-1ß translates into a profound loss of anti-inflammatory TH17 cell functionalities, which can be reversed by anti-IL-1ß treatment. CONCLUSION: IL-1ß signaling determines the differential expression pattern of IL-10, which is necessary and sufficient to induce proinflammatory versus anti-inflammatory TH17 cell functions. Our data introduce TH17 cell subsets as novel players in autoinflammation and thus novel therapeutic targets in autoinflammatory syndromes including other IL-1ß mediated diseases. This demonstrates for the first time alterations in the adaptive immune system in patients with autoinflammatory syndromes.

Prerequisites for Functional Interleukin 31 Signaling and Its Feedback Regulation by Suppressor of Cytokine Signaling 3 (SOCS3).

Interleukin-31 (IL-31) is a T helper type 2 cell-derived cytokine tightly associated with inflammatory skin disorders. IL-31-induced signaling is mediated by a receptor complex composed of oncostatin M receptor ß and the cytokine-specific receptor subunit IL-31Rα, of which there are several isoforms. The latter can be classified as long or short isoforms with respect to their intracellular domain. At present, the signaling capabilities of the different isoforms remain inchoately understood, and potential mechanisms involved in negative regulation of IL-31Rα signaling have so far not been studied in detail. Here, we show that both the long and short isoforms of IL-31Rα are capable of inducing STAT signaling. However, the presence of a functional JAK-binding box within IL-31Rα is an essential prerequisite for functional IL-31-mediated STAT3 signaling. Moreover, both the long and short isoforms require oncostatin M receptor ß for their activity. We also show that IL-31 induces expression of four suppressor of cytokine signaling family members and provide evidence that SOCS3 acts as a potent feedback inhibitor of IL-31-induced signaling. Taken together, this study identifies crucial requirements for IL-31 signaling and shows its counter-regulation by SOCS3.

Human Th2 but not Th9 cells release IL-31 in a STAT6/NF-κB-dependent way.

IL-31, a member of the IL-6 protein family, is one of the latest additions to the list of T cell-derived cytokines. Th2 cells are regarded as a main source of IL-31, which is produced in response to stimulation by IL-4. Because the development of Th9 cells also requires IL-4 as a polarizing cytokine, the current study investigates IL-31 production in human Th9 cells compared with Th2 cells. We found that, although Th9 cells were able to release IL-31 during the first weeks of in vitro polarization, no IL-31 was detected in Th9 cultures after a final restimulation in the absence of polarizing cytokines. We further show that TGF-ß, which is required to obtain Th9 cells in vitro, potently inhibits the release of IL-31 from Th2 cells, whereas IL-33, a cytokine associated with Th2-mediated inflammation, synergizes with IL-4 in inducing IL-31 secretion. To analyze the molecular mechanisms underlying the induction of IL-31, EMSAs, reporter gene assays, and small interfering RNA-based silencing experiments were carried out. We show that STAT6 and NF-κB are central players in mediating IL-31 expression induced by IL-4/IL-33. In addition, we identified a novel NF-κB-binding element within the Il31 promoter that mediates the enhancing effects of IL-33 on IL-4/STAT6-induced IL-31 expression in human Th2 cells. Taken together, this study shows that IL-4 is essential for the production of IL-31, whereas TGF-ß significantly suppresses IL-31 expression at the mRNA and protein levels. As a consequence, in vitro polarized Th2 cells, but not Th9 cells, are able to release IL-31.

STAT6-dependent and -independent mechanisms in Th2 polarization.

Th2 cells play a key role in directing immune responses against helminths. Additionally, Th2 cells are crucial for many types of allergic reactions. Whereas the molecular mechanisms underlying the differentiation of other types of Th cells are well understood, Th2 differentiation is still a controversial topic. IL-4 and its downstream transcription factor signal transducer and activator of transcription (STAT)6 are well-known key mediators in Th2 differentiation. The fact that Th2 cells themselves are the most potent source of IL-4 suggests that additional mechanisms promoting the initiation of Th2 differentiation exist. This article gives an overview on STAT6-dependent and -independent mechanisms involved in the process of Th2 polarization, including Notch, mTORC2, IL-2/STAT5, and Wnt. Furthermore, we emphasize the role of STAT6 not only as a transcriptional activator promoting Th2 development, but also in fine-tuning alternative signaling pathways which are involved in the initiation of Th2 polarization.

Dendritic cells activated by IFN-γ/STAT1 express IL-31 receptor and release proinflammatory mediators upon IL-31 treatment.

IL-31 is a T cell-derived cytokine that signals via a heterodimeric receptor composed of IL-31Rα and oncostatin M receptor ß. Although several studies have aimed to investigate IL-31-mediated effects, the biological functions of this cytokine are currently not well understood. IL-31 expression correlates with the expression of IL-4 and IL-13 and is associated with atopic dermatitis in humans, indicating that IL-31 is involved in Th2-mediated skin inflammation. Because dendritic cells are the main activators of Th cell responses, we posed the question of whether dendritic cells express the IL-31R complex and govern immune responses triggered by IL-31. In the current study, we report that primary human CD1c(+) as well as monocyte-derived dendritic cells significantly upregulate the IL-31Rα receptor chain upon stimulation with IFN-γ. EMSAs, chromatin immunoprecipitation assays, and small interfering RNA-based silencing assays revealed that STAT1 is the main transcription factor involved in IFN-γ-dependent IL-31Rα expression. Subsequent IL-31 stimulation resulted in a dose-dependent release of proinflammatory mediators, including TNF-α, IL-6, CXCL8, CCL2, CCL5, and CCL22. Because these cytokines are crucially involved in skin inflammation, we hypothesize that IL-31-specific activation of dendritic cells may be part of a positive feedback loop driving the progression of inflammatory skin diseases.

IL-37: a new anti-inflammatory cytokine of the IL-1 family.

The IL-1 family of cytokines encompasses eleven proteins that each share a similar ß-barrel structure and bind to Ig-like receptors. Some of the IL-1-like cytokines have been well characterised, and play key roles in the development and regulation of inflammation. Indeed, IL-1α (IL-1F1), IL-1ß (IL-1F2), and IL-18 (IL-1F4) are well-known inflammatory cytokines active in the initiation of the inflammatory reaction and in driving Th1 and Th17 inflammatory responses. In contrast, IL-1 receptor antagonist (IL-1Ra, IL-1F3) and the receptor antagonist binding to IL-1Rrp2 (IL-36Ra, IL-1F5) reduce inflammation by blocking the binding of the agonist receptor ligands. In the case of IL-37 (IL-1F7), of which five different splice variants have been described, less is known of its function, and identification of the components of a heterodimeric receptor complex remains unclear. Some studies suggest that IL-37 binds to the α chain of the IL-18 receptor in a non-competitive fashion, and this may explain some of the disparate biological effects that have been reported for mice deficient in the IL-18R. The biological properties of IL-37 are mainly those of down-regulating inflammation, as assessed in models where human IL-37 is expressed in mice. In this review, an overview of the role of IL-37 in the regulation of inflammation is presented. The finding that IL-37 also locates to the nucleus, as do IL-1α and IL-33, for receptor-independent organ/tissue-specific regulation of inflammation is also reviewed.

Inhibition of suppressive T cell factor 1 (TCF-1) isoforms in naive CD4+ T cells is mediated by IL-4/STAT6 signaling.

The Wnt pathway transcription factor T cell factor 1 (TCF-1) plays essential roles in the control of several developmental processes, including T cell development in the thymus. Although previously regarded as being required only during early T cell development, recent studies demonstrate an important role for TCF-1 in T helper 2 (Th2) cell polarization. TCF-1 was shown to activate expression of the Th2 transcription factor GATA-binding protein 3 (GATA3) and thus to promote the development of IL-4-producing Th2 cells independent of STAT6 signaling. In this study, we show that TCF-1 is down-regulated in human naive CD4(+) T cells cultured under Th2-polarizing conditions. The down-regulation is largely due to the polarizing cytokine IL-4 because IL-4 alone is sufficient to substantially inhibit TCF-1 expression. The IL-4-induced suppression of TCF-1 is mediated by STAT6, as shown by electrophoretic mobility shift assays, chromatin immunoprecipitation, and STAT6 knockdown experiments. Moreover, we found that IL-4/STAT6 predominantly inhibits the shorter, dominant-negative TCF-1 isoforms, which were reported to inhibit IL-4 transcription. Thus, this study provides a model for an IL-4/STAT6-dependent fine tuning mechanism of TCF-1-driven T helper cell polarization.

Food preservatives sodium benzoate and propionic acid and colorant curcumin suppress Th1-type immune response in vitro.

Food preservatives sodium benzoate and propionic acid and colorant curcumin are demonstrated to suppress in a dose-dependent manner Th1-type immune response in human peripheral blood mononuclear cells (PBMC) in vitro. Results show an anti-inflammatory property of compounds which however could shift the Th1-Th2-type immune balance towards Th2-type immunity.

Induction of immune tolerance by oral IVIG.

In the last years evidence has been provided for the importance of B cells in the pathogenesis of rheumatoid arthritis (RA). Several studies have supported the concept that humoral immunity, manifested by the production of autoantibodies, such as rheumatoid factors (RFs), plays a significant role in the course of the disease. Specific targeting of autoantibody-producing B cells, such as RF-producing B cells, should therefore be a promising new approach in the treatment of RA. We used a mouse model to induce human RF responses and asked the question whether oral treatment with the antigen (human IgG) recognized by RFs could induce immune tolerance to RF responses. Balb/c mice were orally treated with polyvalent human IgG before and after immunization with insoluble immune complexes (ICs) that triggered the induction of RFs. Serum titers of RFs were significantly reduced after both primary and booster immunization when human IgG was given as a single oral dose or continuously in drinking water. Continuous treatment with human IgG even prevented booster effects on RFs when treatment started after primary immunization. Treatment with IgG fragments provided evidence that the observed effect of human IgG was mediated by the Fc part and not the Fab part of IgG. Furthermore, transfer of spleen cells obtained from mice after oral treatment with human IgG suppressed RF responses in recipient mice. These data give promising indications that oral human IgG might represent an alternative approach for immunosuppressive B-cell targeted therapies in RA.

Hip and nonvertebral fracture prediction in nursing home patients: role of bone ultrasound and bone marker measurements.

CONTEXT: Absolute fracture risk in nursing home patients is the highest among the communities studied. Screening for high-risk patients in such an environment is usually difficult. OBJECTIVE: The objective was to investigate whether quantitative bone ultrasound measurements and/or markers of bone turnover/metabolism help in predicting which patients will incur hip or nonvertebral fractures. DESIGN, SETTING, AND PARTICIPANTS: In this prospective study, mobile teams enrolled 1664 female patients from 95 nursing homes in Austria. MAIN OUTCOME MEASURES: Calcaneal stiffness (n = 1117), radial speed of sound (SOS) (n = 1332), and phalangeal SOS (n = 1498) measurements were performed at baseline. Serum samples (n = 960) were analyzed for serum calcium and phosphate, 25 hydroxyvitamin D, PTH, osteocalcin, C-terminal telopeptide crosslinks, and osteoprotegerin (OPG). Patients were prospectively followed for hip and other nonvertebral fractures for 2 yr. RESULTS: A total of 117 hip fractures and 269 nonvertebral fractures developed during a mean observation period of 2 yr. Prevalence of vitamin D deficiency and secondary hyperparathyroidism was high. A history of a past fracture was significantly associated with a hazard ratio (HR) of 1.47 (95% confidence interval, 1.01-2.15) and 1.65 (1.26-2.16) for the development of hip and nonvertebral fractures, respectively. Cox regression analysis revealed a multivariate adjusted elevation in both hip [HR 1.30 (1.12-1.43)] and nonvertebral [HR 1.14 (1.02-1.25)] fracture risk for each sd decrease in calcaneal stiffness. Patients in the lowest quartile for calcaneal stiffness Z-score had 2.5 and 1.2 times higher rates of hip and nonvertebral fractures when compared with patients in the highest quartile. Fracture rates were not statistically associated with baseline radial or phalangeal SOS measurements or with serum osteocalcin, C-terminal telopeptide crosslinks, and OPG concentrations. When adjusted for bone mass, higher serum OPG levels were associated with fewer hip as well as nonvertebral fractures [HR 0.85 (0.73-0.99) and 0.89 (0.80-0.99) per increment of 1]. Higher serum phosphate levels indicated an increased hip [HR 1.54 (1.07-2.21)] and nonvertebral fracture risk [HR 1.40 (1.10-1.78) per increase of 1 mg/dl]. Body mass index was protective of hip fractures [HR 0.94 (0.90-0.98) per increase of 1] as well as medication with acetylsalicylic acid [HR 0.59 (0.36-0.95) for hip and 0.72 (0.52-0.99) for nonvertebral fractures]. In contrast, current use of glucocorticoids [HR 5.65 (1.77-18.0)] and opiates [HR 1.85 (1.18-2.92)] exerted a negative effect on prospective hip fracture risk. CONCLUSION: Calcaneal stiffness measurements proved to be useful in predicting hip fractures and to a lesser extent nonvertebral fractures in nursing home residents. Radial and phalangeal bone ultrasound measurements and baseline markers of bone turnover, however, were not indicative of future fracture risk in this population.

Identification of a distal tandem STAT6 element within the CCL17 locus.

The CC chemokine ligand CCL17 is one of the major chemo-attractants for T(H)2 cells. Interleukin-4-induced activation of CCL17 expression was recently demonstrated to result from two STAT6 motifs in the proximal promoter. Here we provide evidence that a distal tandem STAT6 element further elevates expression from the CCL17 locus approximately twofold. This is demonstrated by reporter gene assays using different fragments of the CCL17 promoter and the region 2.5 kb upstream from the transcriptional start site. By electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation experiments, we show that STAT6 binds to the motifs in vitro and in vivo, respectively. Insertion of nucleotide exchanges into the STAT6 core motifs results in diminished promoter activation and abrogated STAT6 binding, as demonstrated by reporter gene and EMSA studies. Collectively these data reveal an additional element involved in the regulation of CCL17 expression.

Value of 3-dimensional transrectal/transvaginal sonography in diagnosis of distal ureteral calculi.

OBJECTIVE: In a prospective study, the feasibility of 3-dimensional (3D) transrectal/transvaginal sonography in comparison with transabdominal sonography and intravenous urography (IVU) in identifying distal ureteral calculi was evaluated. METHODS: Sixty-two patients in the urologic clinic with clinical suspicion of distal ureteral calculi were included. The patients consisted of 44 men and 18 women with a mean age +/- SD of 44 +/- 17 years. These patients underwent 3D transrectal/transvaginal sonography, transabdominal sonography with IVU, and, finally, ureterorenoscopy. RESULTS: Fifty-nine patients were confirmed to have distal ureteral calculi on the basis of urologic intervention (ureterorenoscopy). Three patients had a spontaneous stone passage immediately after imaging completion. The median size of the calculi was 3.7 +/- 2.00 mm. Transabdominal sonography detected 34 of the 62 patients with calculi (sensitivity, 55%). The median size of the calculi was calculated as 5.0 +/- 2.4 mm. The examination time was 6.5 +/- 2.7 minutes. Intravenous urography detected 44 of the 62 patients with ureterolithiasis (sensitivity, 71%). Herein, the median stone size was measured as 3.9 +/- 1.9 mm, and the examination time was 38 +/- 17 minutes. The combination of transabdominal sonography and IVU in visualization of ureterolithiasis raised the sensitivity to 81% (50 of 62 patients). Three-dimensional transrectal/transvaginal sonography showed ureterolithiasis in all 62 patients confirmed to have distal ureteral calculi (sensitivity and specificity, 100%). The median size of the calculi was calculated as 4.4 +/- 2.2 mm, and the examination took 1.9 +/- 0.6 minutes. CONCLUSIONS: The data in our prospective study show that transrectal/transvaginal sonography with 3D image assessment is superior to IVU and abdominal sonography for diagnosing distal ureteral calculi.

Three-dimensional ultrasonography of the urinary bladder: preliminary experience of assessment in patients with haematuria.

OBJECTIVE: To assess the value of three-dimensional (3D) vs two-dimensional (2D) ultrasonography (US) in the diagnostic evaluation of the urinary bladder in patients with haematuria. PATIENTS AND METHODS: In all, 42 patients with painless haematuria and/or irritative voiding symptoms were examined with 2D- and 3D-US. US was done with an Acuson Sequoia unit (Siemens Medical Sol. Mountain View, CA, USA) and the Perspective(R) 3D technique, to assess the presence of bladder lesions, including bladder cancer, bladder wall hypertrophy with trabeculation and diverticula, mucosal bladder folds or re-growth of the prostate mimicking a bladder tumour. The imaging findings were compared with cystoscopy and/or bladder biopsy. RESULTS: In 21 of the 42 patients (50%) cystoscopy with bladder biopsy revealed bladder cancer. Overall, 3D-US gave a correct diagnosis for 36 of 42 patients (86%). All 21 bladder cancers were correctly diagnosed, and 15 (71%) of the 21 benign bladder lesions were correctly identified. By contrast, 2D-US findings gave suspected bladder cancer in all patients. CONCLUSIONS: 3D-US is significantly more accurate than standard 2D-US in the diagnostic evaluation of patients with haematuria. Thus, this diagnostic technique might be useful for routine evaluation of the urinary bladder.

Type 2 diabetes mellitus in nursing home patients: effects on bone turnover, bone mass, and fracture risk.

CONTEXT: Fractures are a major health burden in elderly institutionalized persons. Type 2 diabetes mellitus (DM) has a high prevalence in nursing home patients and has been associated with positive effects on bone mass in younger, community-dwelling elderly. OBJECTIVE: The objective of this study was to investigate whether type 2 DM affects bone mass, bone turnover, or prospective fracture rates in frail, elderly women living in nursing homes. DESIGN, SETTING, AND PARTICIPANTS: This study was a prospective cohort of 583 patients with type 2 DM and 1081 control (CTR) individuals above age 70 recruited from 95 nursing homes in Austria. Patients were enrolled and followed up by mobile study teams. MAIN OUTCOME MEASURES: We performed quantitative bone ultrasound measurements at the calcaneus, radius, and proximal third phalanx, measurements of quadriceps strength, and biochemical parameters of mineral metabolism and bone turnover. Patients were prospectively followed for hip and other nonvertebral fractures over 2 yr. RESULTS: Patients with type 2 DM had significantly higher age-, weight-, and mobility score-adjusted calcaneal stiffness (P < 0.0001), radial speed of sound (P < 0.005), and phalangeal speed of sound (P < 0.05) measurements when compared with CTRs. Mean serum PTH (-20.7%) and osteocalcin levels (-22.3%) were significantly lower (both P < 0.0001) in patients with treated type 2 DM despite comparable low serum 25-hydroxyvitamin D levels and slightly higher adjusted total serum calcium levels compared with CTRs. Important independent determinants of bone turnover in both patient groups were PTH, creatinine clearance, alanine aminotransferase, as well as glycosylated hemoglobin levels, together accounting for 30-40% of its variance. A total of 110 hip fractures occurred during the observation period, corresponding to a hip fracture rate of 3.1% (in CTRs) and 3.4% (in type 2 DM) per 100 patient years; this was not significantly different for CTRs and diabetics. CONCLUSIONS: Decreased PTH levels and higher levels of glycemia independently contribute to lower bone turnover in elderly nursing home patients with type 2 DM. Despite higher bone mass and lower bone turnover, hip fracture risk is comparable with women without DM.

Long-term persistence of anti-factor VIII antibody-secreting cells in hemophilic mice after treatment with human factor VIII.

The analysis of anti-factor VIII (FVIII) antibody-secreting cells (ASC) at different anatomic sites provides valuable information about the nature of the anti-FVIII immune response in hemophilic mice after treatment with human FVIII. An Elispot system is described that is suitable for analyzing frequencies and IgG subclasses of anti-FVIII ASC at the single-cell level. Hemophilic mice were treated with four doses of FVIII. Anti-FVIII antibodies in blood as well as anti-FVIII ASC in spleen and bone marrow were analyzed after each dose of FVIII and subsequently up to 22 weeks after termination of the FVIII treatment. Anti-FVIII ASC first appeared in the spleen where they were detectable after two intravenous doses of FVIII. Their appearance correlated with that of anti-FVIII antibodies in blood plasma. Anti-FVIII ASC in bone marrow were detectable after three doses of FVIII and were probably cells that initially formed in the spleen and subsequently migrated to the bone marrow. Whereas the frequency of anti-FVIII ASC in the spleen increased up to the fourth dose of FVIII and declined thereafter, in the bone marrow it remained constant for up to at least 22 weeks after the termination of the FVIII treatment. Titers of anti-FVIII antibodies in blood plasma increased up to the fourth dose of FVIII, then remained high constantly for 14 weeks and decreased but the antibodies were still detectable for up to at least 22 weeks after the fourth dose of FVIII. The IgG-subclass distribution of anti-FVIII ASC was similar in spleen and bone marrow and matched the subclasses of anti-FVIII antibodies in blood plasma indicating that both organs contribute to circulating antibodies in the blood.