Nonuremic calciphylaxis precipitated by teriparatide [rhPTH(1-34)] therapy in the setting of chronic warfarin and glucocorticoid treatment.

Calciphylaxis occurs rarely in the absence of end stage renal disease. Predisposing factors for nonuremic calciphylaxis (NUC) include hyperparathyroidism, coagulopathies, connective tissue disease, liver disease, glucocorticoid use, and malignancy. Warfarin can facilitate vascular calcification by reducing vitamin K-dependent carboxylation of matrix-Gla proteins. An 86-year-old Caucasian woman with a history of polymyalgia rheumatica, two spontaneous deep venous thromboses (DVTs) and multiple fractures was treated with calcium, vitamin D, prednisone, and warfarin. The patient's low bone density was treated initially with estrogen, then oral bisphosphonate, which was discontinued due to upper gastrointestinal symptoms. Nasal calcitonin was initiated. After 10 years of calcitonin treatment, she was changed to teriparatide. Two months after initiating teriparatide, she developed lower extremity edema and painful erythematous nodular lesions on her calves bilaterally, that progressed to necrotic ulcers despite antibiotic therapy. Biopsy of the lesions showed calcification in the media of small blood vessels and subcutaneous fat with fat necrosis, consistent with calciphylaxis. Teriparatide was discontinued. Aggressive wound care, antibiotics, and intravenous zoledronic acid were initiated. With cessation of teriparatide therapy and intensive wound care, the patient's lesions resolved over 8 months. We report the first case of NUC precipitated by teriparatide therapy. Our patient had multiple underlying predisposing factors including a connective tissue disorder, glucocorticoid therapy, warfarin use, and possible underlying coagulopathy given her history of multiple DVTs. In such patients, alternative osteoporosis therapies may be preferred.

Cuff-less blood pressure estimation using wrist photoplethysmography.

One of the most promising and at the same time rapidly growing sectors in healthcare is that of wearable medical devices. Population ageing constantly shifts towards a higher number of senior and elderly people with increased prevalence of chronic diseases often requiring long-term care and a need to decrease hospitalization time and cost. However, today most of the devices entering the market are not standardized nor medically approved, and they are highly inaccurate. In this work we present a system and a method to provide accurate measurement of systolic and diastolic blood pressure (BP) based solely on wrist photoplethysmography. We map morphological features to BP values using machine learning and propose ways to select high quality signals leading to an accuracy improvement of up to 33.5%, if compared against no signal selection, a mean absolute error of 1.1mmHg in a personalized scenario and 8.7mmHg in an uncalibrated leave-one-out scenario.

High prevalence of metabolic syndrome and cardiovascular risk factors in men with ankylosing spondylitis on anti-TNFalpha treatment: correlation with disease activity.

OBJECTIVE: Ankylosing spondylitis (AS) may be associated with an increased risk for cardiovascular diseases (CVD). We investigated the prevalence of cardiovascular risk factors and metabolic syndrome (MetS) in men with AS and assessed any correlation with AS-related factors. METHODS: This was a cross-sectional study of 63 men with AS, median age 40 (19-69) years, and 126 age-matched controls. Patients were on anti-TNFalpha treatment because of considerable disease activity at some time during the course of the disease. MetS was assessed according to the modified National Cholesterol Education Program Adult Treatment Panel III criteria. The risk for CVD event within the next 10 years was estimated using the Framingham equation. RESULTS: Patients had lower high-density lipoprotein cholesterol (HDL-C) (p<0.001), higher systolic (p=0.001) and diastolic (p<0.01) blood pressure compared with controls. The prevalence of the MetS was higher in patients compared to controls (34.9% vs. 19.0%; p<0.05). AS patients with MetS were older (p<0.01), with higher Framingham risk score (p=0.001), had longer disease duration (p<0.05) and higher BASDAI (5.1 vs. 3.7; p<0.05) than those without MetS, while both BASFI and CRP had an inverse correlation with HDL-C levels. CONCLUSIONS: Men with AS have a higher prevalence of cardiovascular risk factors and MetS compared with controls. The presence of MetS was associated with increased 10 year CVD risk in these patients. The association of AS disease activity with MetS suggests that CVD in AS patients may, at least in part, be attributed to the inflammatory burden of the disease.

Interleukin 2 production and interleukin 2 receptor expression by human immature leukemic T cells.

In order to determine the role of interleukin 2 (IL2) on the proliferation of leukemic cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) we studied the production of IL2, the function of IL2 receptors (IL2R) expressed on T-ALL cells and their IL-2-dependent in vitro proliferation. Leukemic cells from six out of 17 T-ALL/T-cell non-Hodgkin's lymphoma patients with a prothymocyte (stage I) or a mature thymocyte (stage III), but not with a common thymocyte (stage II) phenotype, could proliferate, in a dose-dependent manner, in response to recombinant IL2 (rIL2) and anti-Tac and TU27 moAbs as well as polyclonal anti-IL2 purified immunoglobulin G could inhibit this IL2-induced cell proliferation. Both crude or/and Amicon-concentrated media conditioned by T-ALL cells from 10 out of 13 tested patients contained IL2 activity as assessed by colorimetric biological and immunoenzymatic assays; this biologic activity was due to a 14.5 kDa molecule adsorbed by anti-IL2 antibodies in an immunoaffinity assay. Although less than 10% of fresh leukemic cells expressed IL2R alpha (Tac) chain, a 24 h cell incubation in the absence of any mitogenic stimulation induced IL2R alpha chain expression in five out of 13 patients (11-83% Tac+ cells). Morever, Tac mRNA transcripts could be detected in fresh cells from all 10 patients tested. Staining of fresh leukemic cells with an IL-2R beta-chain-specific monoclonal antibody and flow cytometry analysis revealed that 4-13% of leukemic cells were positive. Binding experiments with 125I-rIL2 showed a small number of high affinity IL2R on fresh cells from three T-ALL patients (114-200 sites/cell, dissociation constant = 101-181 pm). Finally, antibodies against IL2R alpha, IL2R beta and IL2 could inhibit both IL2 driven and spontaneous cell proliferation of most patients' T-ALL cells, although in some cases an heterogenous pattern of inhibition was observed. Taken together, these findings strongly suggest that an IL2/IL2R-dependent mechanism could be involved in the proliferation of some T-ALL cells.

Intracytoplasmic detection of the Tac (p55) chain of interleukin-2 receptor in pre-B leukemic cells associated with a constitutive expression of Tac mRNA.

The pre-B Reh-6 leukemic cells do not express membrane interleukin-2 (IL-2)-R alpha (Tac or p55) chain; however, their incubation with PMA induces the expression of both high and low affinity IL-2-R. Northern analysis of nonstimulated Reh-6 as well as leukemic cells from patients with acute B cell precursor lymphoblastic leukemia displayed a constitutive expression of p55 mRNA transcripts, which could be enhanced by PMA. Both actinomycin-D and cycloheximide could abrogate PMA-induced p55 membrane expression, suggesting the need for de novo mRNA and protein synthesis. The increased PMA-induced p55 mRNA accumulation was an early event (4 hr) and could be enhanced, specifically, by rIL-2 because anti-Tac moAb inhibited this rIL-2-mediated effect. Immunofluorescence and cross-linking studies using 125I-rIL-2 failed to reveal membrane-associated p55 protein on both Reh-6 and patients' leukemic cells. Conversely, immunogold staining and electron microscopy studies, revealed p55 immunoreactive molecules in the cytoplasm but not in the nucleus of all Reh-6 cells. Using a sensitive EIA, p55 molecules could be detected in cell lysates but not the culture supernatants of Reh-6 cells, suggesting that p55 was not released into the culture medium. These results indicate that constitutively expressed p55 mRNA on pre-B leukemic cells is translated into a relative immunoreactive protein that cannot be expressed on cell surface for unknown, yet, reasons.

Microplate-array diagonal-gel electrophoresis (MADGE) and melt-MADGE: tools for molecular-genetic epidemiology.

Microplate-array diagonal-gel electrophoresis (MADGE) was invented for molecular-genetic epidemiological studies. It combines direct compatibility with microplates, convenient polyacrylamide-gel electrophoresis and economy of time and reagents at minimal capital cost, and enables one user to run up to several-thousand gel lanes per day for the direct assay of single-base variations. Melt-MADGE adds temporal-thermal-ramp apparatus to achieve similar throughput for de novo mutation scanning.

Polymer-nanotube composite mats with improved field emission performance and stability.

The results of electron field emission from single walled carbon nanotubes (SWCNTs) mats deposited on different composite films of SWCNTs and poly(3-octylthiophene) (P3OT) semiconducting polymer are presented. Three different structures were tested: (a) dense and sparse SWCNT mats on n+ -Si; (b) SWCNT mats on composite films with different SWCNT-P3OT ratios; (c) composite films with different SWCNT-P3OT ratios on n+ -Si. The experiments show that there is a critical SWCNT-P3OT concentration in which the field emission stability of SWCNT mats is remarkably improved with a small reduction in the emission threshold compared to the optimum pristine SWCNT film. The contribution of the composite film morphology as well as the role of polymer-nanotube interaction on the emission performance are evaluated. The physical mechanism behind the stability of composite field emitters is also discussed.

Metabolic syndrome is common among middle-to-older aged Mediterranean patients with rheumatoid arthritis and correlates with disease activity: a retrospective, cross-sectional, controlled, study.

OBJECTIVES: Patients with rheumatoid arthritis have an increased risk for cardiovascular disease (CVD). The prevalence of metabolic syndrome (MetS)-a major contributor to CVD-in a cohort of patients with rheumatoid arthritis and its relationship with rheumatoid arthritis related factors is investigated here. METHODS: 200 outpatients with rheumatoid arthritis (147 women and 53 men), with a mean (standard deviation (SD)) age of 63 (11) years, and 400 age and sex-matched controls were studied. MetS was assessed according to the adult treatment panel III criteria and rheumatoid arthritis disease activity by the disease activity score of 28 joints (DAS28). A standard clinical evaluation was carried out, and a health and lifestyle questionnaire was completed. RESULTS: The overall prevalence of MetS was 44% in patients with rheumatoid arthritis and 41% in controls (p = 0.5). Patients with rheumatoid arthritis were more likely to have low high-density lipoprotein cholesterol compared with controls (p = 0.02), whereas controls were more likely to have increased waist circumference or raised blood pressure (p = 0.001 and 0.003, respectively). In multivariate logistic regression analysis adjusting for demographics and rheumatoid arthritis treatment modalities, the risk of having moderate-to-high disease activity (DAS28>3.2) was significantly higher in patients with MetS compared with those with no MetS components (OR 9.24, 95% CI 1.49 to 57.2, p = 0.016). CONCLUSION: A high, albeit comparable to the control population, prevalence of MetS was found in middle-to-older aged patients with rheumatoid arthritis. The correlation of rheumatoid arthritis disease activity with MetS suggests that the increased prevalence of coronary heart disease in patients with rheumatoid arthritis may, at least in part, be attributed to the inflammatory burden of the disease.

Problems related to the interpretation of autoradiographic data on gene expression using common constitutive transcripts as controls.

The 28S rRNA, a ribosomal RNA, and the ACTB and GAPD mRNAs, coding respectively for beta-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are frequently presented as controls of modulated gene expression. These transcripts were quantified by replicate slot-blot autoradiography and image analysis in mammary epithelial cells and fibroblasts from breast tissues. Each cell-type group comprised strains with different pathological backgrounds, growth rates, antigenic phenotypes and culture histories. The effects of a differentiating agent (cholera toxin) and/or a tumor promoter (12-O-tetradecanoyl-phorbol-13-acetate) were also examined. Despite the impression that visual examination of autoradiographs might create, image analysis suggests that 28S rRNA, ACTB and GAPD are substantially and independently influenced by the above biological factors and by the drugs. Therefore, these transcripts represent specifically regulated cellular activities and may not be taken as alternative indicators of the overall transcription rate or of the amount of material being examined. Instead, such nonspecific variation may be accurately measured and removed from quantitative data using a principal component function. A methodology that allows comparison of expression (or amplification) patterns between genes, between experiments or, even, between laboratories is presented with an example of quantification of transcripts related to cell-growth, differentiation, signaling and cancer.

[Histological study of the remains of periodontal membrane at the alveolar wall after tooth extraction]. / Histologische Untersuchungen über Desmodontreste an Alveolarwänden nach einer Zahnextraktion.

Histological study of 40 alveolar wounds of altogether 5 dogs have given evidence to the fact that remains of the periodontal membrane could remain in the apical area of the alveoli in their whole width. In the middle part of the alveoli, remains of it are found in a thin layer. In the marginal area, bone can be found lying open with torn remnants, which are lying free in the coagulum. In these adhering rests of the periodontal membrane plenty of dynamic cells as well as dilated vessels can be observed. All these facts lead to the idea of considering the part that the remains of the periodontal membrane may possibly play in the beginning process of alveolar wound healing.

Evaluation of quantitative variation in gene expression.

We investigate the behaviour of the gene-expression rate as a statistical variable using autoradiographic data for 39 transcripts from a heterogeneous set of 80 breast-tissue cultures. Despite standardization, the data distributions of all transcripts showed intervals of normality and intervals of systematic departure from normality which most frequently resulted in a significant skewness and/or kurtosis. Non-normal shapes are attributed to modulation of gene expression. This statistical particularity creates difficulties in the evaluation of differences among specimens. Using classical parametric and non-parametric procedures for normal and non-normal variation, respectively, we demonstrate that large differences in optical density are neither necessary nor sufficient for associating expression rates with biological factors. The transcripts coding for the metalloprotease stromelysin-3 (ST3) and for the receptor to insulin-like growth factors (IGFR) are used as examples and their variation is presented in detail. ST3 expression appeared to be specifically associated with mammary stroma fibroblasts derived from post-radiation fibrosis lesions. IGFR was expressed at higher rates in mammary gland and skin fibroblasts than in mammary epithelial cells and was subject to frequent and strong modulation.

Quantitative variation of proto-oncogene and cytokine gene expression in isolated breast fibroblasts.

Transcripts coding for transcription factors (RB, P53, FOS, MYC, MYB, ERBA, REL), growth factors (FGF1, FGF2, INT2, TGFA, TGFB, PDGF, IGF1, IGF2), interleukins, (IL1, IL2, IL3, IL4, IL6, TNF), growth-factor receptors or cytosolic protein kinases (RAF, PIM, FES, MET, SRC, ROS, TRK, KIT, CSFR, IGFR, PDGFR, EGFR, NEU) were quantified in cultured human mammary fibroblasts from normal tissues, benign tumours, carcinomas and post-radiation fibrosis lesions by slot-blot autoradiography and image analysis. The effects of a differentiating agent (cholera toxin) and of a tumour promoter (12-O-tetradecanoyl-phorbol-13-acetate) were also examined. The drugs modulated the levels of the anti-oncogene transcripts (RB, P53) and of ERBA, REL, RAF, MET, ROS, TRK, CSFR, EGFR, NEU, FGF1, INT2, IGF1, IL1, IL2, IL4 and IL6. Apart from this variation, there were multiple differences in gene expression among normal and pathological cells (concerning all but P53, TGFB and interleukin transcripts) and between sub-types defined by the presence of alpha-sm-actin (myofibroblasts) or EDB-fibronectin (RAF, ROS, FES, KIT, IGFR, NEU, INT2, TGFB, PDGF, IGFs, ILs). It appears, therefore, that mammary stroma progress irreversibly along with the epithelium during tumoral development, and that breast cancer is not only a multi-gene but also a multi-tissue phenotype.

Discrimination of fibroblast subtypes by multivariate analysis of gene expression.

Fibroblasts and myofibroblasts from normal, fibrotic or tumoral breast tissues present multiple quantitative differences in gene expression even when grown in isolation. We were therefore prompted to investigate whether one could recognize various subtypes by their constitutive-gene expression profile. Quantitative autoradiographic data for 34 constitutively expressed transcripts were submitted to multivariate analysis of variance, followed by discriminant analysis and single linkage cluster analysis. Models assuming up to 8 putative fibroblast subtypes (among fibroblasts or myofibroblasts from breast skin, normal mammary stroma, tumor-adjacent "normal" stroma, post-radiation fibrosis lesions and benign or malignant tumors) and an epithelial-cell group used as an internal control resulted in 100% correct classification. Myofibroblasts from various origins clustered close to, although distinctly apart from, their corresponding alpha-smooth-muscle-actin-negative counterparts. Malignant tumor fibroblasts were phenotypically more distant from normal cells compared with other pathological types. Our results support the hypothesis of co-adaptive transformation of stromal and epithelial tissues during breast tumoral development and suggest that different types of fibroblasts give rise to different types of myofibroblasts. Discriminant analysis of quantitative molecular variation may be considered for the development of a powerful artificial-intelligence method for cell typing and should be particularly useful when no reliable discrete molecular markers are available.

Co-adaptation of Escherichia coli and coliphage lambda vir in continuous culture.

Populations of the bacterium Escherichia coli and of its phage lambda vir appeared to equilibrate in continuous cultures. The bacterial end-populations were heterogeneous in respect of their resistance to lambda vir and their ability to utilize maltose. The most competitive of the selected bacteria were mutants which had a reduced rate of synthesis of lambda-receptor so as to become highly, but not totally, resistant to the phage. The coexisting phage had an increased affinity for the receptor and an altered antigenic specificity, suggesting adaptation of its adsorption site in response to the evolution of resistance in the bacteria.

[Conservative therapy of mandibular fractures in children with operatively secured splints]. / Zur konservativen Therapie der Uterkieferfrakturen bei Kindern mit operativ befestigten Schieneverbänden.

In the treatment of mandibular fractures in children offering no adequate possibility for attaching splints to the teeth, we use prosthetic splints fixed with four circumferential wires. This monomaxillary fixation is sufficient to stabilize fractures of the mandibular anterior segment. Fractures situated distal to the canines require additional intermaxillary fixation, i.e. zygomaticomaxillary suspension of the mandible. When functional therapy is needed at the same time, we use an activator instead of the prosthetic splint. 59 patients treated in this way were reevaluated within a period of 10 years. The results of this reevaluation are reported.

[Osprovit: hydroxylapatite ceramic for the reconstruction of blow-out fractures of the orbital floor]. / Osprovit. Plakidia hydroxylapatite gia ten apokatastase syntriptikou katagmatos tou edaphous tou ophthalmikou kogchou.

Common polymeric implantations, in use for the orbital floor reconstruction, form usually a surrounding fissural space and thus tend to dislocations. With the conservative methods that have been available up to now, the Dura Mater cannot be definitely sterilised so that a transmission of a virus may be possible, which causes the Creutzfeld-Jacob Encephalopathy. As an appropriate alternative-both for primary and secondary reconstruction of the orbital floor-an implantation of dense hydroxylapatite ceramics (Osprovit), anatomically shaped, is indicated. According to x-ray techniques of evaluation it presents itself without artifacts. It does not dislocate because it coalesces physiologically with the surrounding tissues and because of its biocompatibility, it is undoubtedly suitable as a permanent implantation. Post-operative examinations of 20 patients showed neither enophthalmus nor diplopia. All patients were without subjective symptoms.