Search

Ischemic heart disease still represents the leading cause of death in the western world despite a decrease of mortality in the last decade. For the diagnostics of coronary artery morphology, invasive coronary angiography represents the gold standard. Nevertheless, in recent years the importance of functional diagnostics of the coronary arteries has increased and various imaging procedures for the measurement of fractional flow reserve (FFR) during coronary angiography were established and recommended for ischemia testing in the actual guidelines on myocardial revascularization.Imaging modalities for diagnostics of the functional relevance of coronary artery disease include stress echocardiography, magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT), and positron emission tomography (PET). These procedures enable advanced risk stratification and therapy guiding in patients with suspected or known coronary artery disease. In future algorithms, hybrid imaging may facilitate the determination of anatomical and functional aspects after only one investigation.In the present article, the role of ischemia testing is compared with morphological methods for the diagnosis of coronary artery disease, individual risk stratification, and therapy guiding.

Objective: To study the relationship of area- and volumetric-based visceral and subcutaneous adipose tissue (VAT and SAT) by MRI and their ratio in subjects with impaired glucose metabolism from the general population.Methods: Subjects from a population-based cohort with established prediabetes, diabetes and healthy controls without prior cardiovascular diseases underwent 3 T MRI. VAT and SAT were assessed as total volume and area on a single slice, and their ratio (VAT/SAT) was calculated. Clinical covariates and cardiovascular risk factors, such as hypertension and glycemic state were assessed in standardized fashion, Univariate and adjusted analyses were conducted.Results: Among 384 subjects (age: 56.2 +/- 9.2 years, 581% male) with complete MRI data available, volumetric and single-slice VAT, SAT and VAT/SAT ratio were strongly correlated (all >r = 0.89), Similarly, VAT/SAT(volum)(e). ratio was strongly correlated with VAT(volume), but not with SAT (r = 0.72 and r = -0.21, respectively). Significant higher levels of VAT, SAT and VAT/SAT ratio were found in subjects with impaired glucose metabolism (all p

Aims To characterize changes in the myocardium in subjects with prediabetes, diabetes, and healthy controls with preserved left ventricular ejection fraction (LVEF) by using cardiac magnetic resonance imaging (CMR) in a sample from the general population.Methods and results Subjects without history of cardiovascular disease and preserved LVEF but established diabetes, prediabetes, and controls from a population-based cohort underwent contrast-enhanced CMR. Obtained parameters included left ventricular (LV) function and morphology, late gadolinium enhancement as well as T1-mapping and derivation of extracellular volume fraction (ECV) by modified Look-Locker inversion recovery for diffuse fibrosis in a subset of patients. Fibrosis volume and cell volume were calculated and LV remodelling index was calculated by dividing the LV mass by its end-diastolic volume. Among 343 subjects (56.1 +/- 9.2 years, 57% males), 47 subjects were classified as diabetes, 78 as prediabetes, and 218 as controls. Haematocrit values and thus ECV parameters were available in 251 subjects. LV remodelling index was significantly higher in participants with prediabetes and diabetes, independent of body mass index (BMI), hypertension, age, and sex. ECV was decreased in subjects with prediabetes and diabetes compared with healthy controls (23.1 +/- 2.4% and 22.8 +/- 3.0%, both P < 0.007). In contrast, cell volume was significantly higher in subjects with prediabetes and diabetes as compared with controls (109.1 +/- 23.8 and 114.9 +/- 32.3 mL vs. 96.5 +/- 26.9 mL, both P < 0.03, respectively). However, differences in ECV and cell volume attenuated after the adjustment for cardiometabolic risk factors, including age, sex, BMI, and hypertension.Conclusion Subjects with prediabetes and diabetes but preserved LVEF had higher LV remodelling indices, suggesting early detectable changes in the disease process, while diffuse myocardial fibrosis appears to be less relevant at this stage.

OBJECTIVES: To compare the correlations of MRI-derived adipose tissue measurements and anthropometric markers, respectively, with prevalent hypertension in a community-based sample, free of clinical cardiovascular disease. METHODS: MRI-derived adipose tissue measurements were obtained in 345 participants (143 women; age 39-73 years) of the KORA FF4 survey from Southern Germany using a 3-Tesla machine and included total adipose tissue (TAT), visceral adipose tissue (VAT), subcutaneous adipose tissue (SCAT), hepatic fat fraction (HFF), pancreatic fat fraction (PFF) as well as pericardial adipose tissue (PAT). In addition, the anthropometric markers body mass index, waist circumference, hip circumference, waist-hip ratio (WHR) and waist-height ratio (WHtR) as well as blood pressure measurements were obtained. RESULTS: The prevalence of hypertension was 33.6% (women: 28%, men: 38%). VAT and PAT had the highest area under the curve (AUC) values for identifying individuals with prevalent hypertension (AUC: 0.75; 0.73, respectively), whereas WHtR and waist circumference were best performing anthropometric markers (AUC: 0.72; 0.70, respectively). A 1SD increment of TAT was associated with the highest odd for hypertension in the age-adjusted and sex-adjusted model (OR = 2.20, 95% CI 1.67-2.91, P

The purpose of this paper is to review the current role of contrast-enhanced cardiac computed tomography (CT) in the peri-procedural planning of mitral valve (MV) repair. Cardiac CT is increasingly implemented in the peri-interventional management of patients undergoing MV repair or MV replacement due to its widespread availability and its ability to provide detailed information on the complex cardiac and valvular anatomy. The complex anatomy of the MV challenges the management of minimally invasive MV repair with respect to device sizing and procedural planning. Advances in CT have enabled cardiac CT to provide critical information for the pre-procedural planning and post-procedural follow-up of MV repair. Therefore, it represents a key element in the improvement of the post-procedural outcome, the efficiency of implanted devices, and the prevention as well as advanced diagnostics of post-procedural complications. However, particular expertise is required to select adequate imaging protocols, perform comprehensive post-processing features, and to achieve specific quantitative image evaluation.

Ischemic heart disease still represents the leading cause of death in the western world despite a decrease of mortality in the last decade. For the diagnostics of coronary artery morphology, invasive coronary angiography represents the gold standard. Nevertheless, in recent years the importance of functional diagnostics of the coronary arteries has increased and various imaging procedures for the measurement of fractional flow reserve (FFR) during coronary angiography were established and recommended for ischemia testing in the actual guidelines on myocardial revascularization.Imaging modalities for diagnostics of the functional relevance of coronary artery disease include stress echocardiography, magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT), and positron emission tomography (PET). These procedures enable advanced risk stratification and therapy guiding in patients with suspected or known coronary artery disease. In future algorithms, hybrid imaging may facilitate the determination of anatomical and functional aspects after only one investigation.In the present article, the role of ischemia testing is compared with morphological methods for the diagnosis of coronary artery disease, individual risk stratification, and therapy guiding.

Advances of computational sciences over the last decades have enabled the introduction of novel methodological approaches in biomedical research. Acquiring extensive and comprehensive data about a research subject and subsequently extracting significant information has opened new possibilities in gaining insight into biological and medical processes. This so-called big data approach has recently found entrance into medical imaging and numerous epidemiological studies have been implementing advanced imaging to identify imaging biomarkers that provide information about physiological processes, including normal development and aging but also on the development of pathological disease states. The purpose of this article is to present existing epidemiological imaging studies and to discuss opportunities, methodological and organizational aspects, and challenges that population imaging poses to the field of big data research.

“Pomegranate has potent antioxidant and anti-inflammatory activities. We therefore investigated if daily consumption of pomegranate extract (PomX) or juice (PJ) for 12 weeks benefited skin health and aging. Additionally, we explored possible mechanisms of pomegranate impact on the skin: microbial composition, expression of epidermis-related genes, and levels of circulating cytokines/chemokines. 74 healthy women (age 36 ± 4) were enrolled and randomly assigned to consume PomX (N=25), PJ (N=24) or matched placebo drink (N=25) for 12 weeks. The minimal erythema dose (MED) was assessed following exposure of inner arm to UVB at baseline and after 12 weeks of Pom consumption. MED was increased significantly in both PomX and PomJ consuming groups compared to placebo. Skin swipes of inner arm were collected for 16S rRNA bacterial sequencing. At the genus level the abundance of 6 and 4 genera were changed significantly by PomX and PJ respectively compared to placebo while alpha and beta diversity were not significantly affected by PomX or PJ. Skin tape expression of inner arm epidermal genes for inflammatory- (SMAD3, TGFβ, IL1β) and skin aging related genes - (MMP1, Stratifin, IGF1R) were not changed by PomX or PJ. Circulating cytokine/chemokine (TGFα, IFNγ, IL10, IL12-p40, IL12-P70, IL1β, IL6, IL8, MCP1, TNFα and VEGF) were not affected by PomX or PJ. Conclusion: Pomegranate consumption resulted in increased MED indicating its photoprotective potential. Further studies are needed to evaluate the association of pom-induced changes in the skin microbiome with its photoprotective potential in normal and aging skin.”

The taxonomy of Echinacea is based on morphological characters and has varied depending on the monog- rapher. The genus consists of either nine species and four varieties or four species and eight varieties. We have used amplified fragment length polymorphisms (AFLP) to assess genetic diversity and phenetic relationships among nine species and three varieties of Echinacea (sensu McGregor). A total of 1086 fragments, of which approximately 90% were polymorphic among Echinacea taxa, were generated from six primer combinations. Nei and Li's genetic distance coefficient and the neighbor-joining algorithm were employed to construct a phenetic tree. Genetic distance results indi - cate that all Echinacea species are closely related, and the average pairwise distance between populations was approxi- mately three times the intrapopulation distances. The topology of the neighbor-joining tree strongly supports two major clades, one containing Echinacea purpurea, Echinacea sanguinea, and Echinacea simulata and the other containing the remainder of the Echinacea taxa (sensu McGregor). The species composition within the clades differs between our AFLP data and the morphometric treatment offered by Binns and colleagues. We also discuss the suitability of AFLP in determining phylogenetic relationships. Resume : La taxonomie du genre Echinacea est basee sur des caracteres morphologiques et varie d'un auteur a un autre. Le genre comprend soit neuf especes et quatre varietes ou encore quatre especes et huit varietes. Les auteurs ont employe le polymorphisme de longueur des fragments amplifies (AFLP) pour mesurer la diversite genetique et les rela- tions phenetiques parmi les neuf especes et trois varietes d' Echinacea (d'apres McGregor). Un total de 1086 fragments, dont environ 90 % s'averaient polymorphes au sein du genre Echinacea, ont ete produits a l'aide de six combinaisons d'amorces. Le coefficient de distance genetique de Nei et Li et l'algorithme NJ ont ete employes pour produire un arbre phenetique. La mesure des distances genetiques suggere que toutes les especes du genre Echinacea sont tres ap- parentees et la distance moyenne entre paires de populations etait environ trois fois plus grande que les distances intra- populations. La topologie de l'arbre NJ supporte nettement deux clades principaux, l'un regroupant l' E. purpurea ,l 'E. sanguinea et l'E. simulata tandis que l'autre regrouperait les autres especes telles que definies par McGregor. Le re - groupement d'especes, au sein de clades, suggere par les donnees AFLP differe de celui obtenu par Binns et al., ce dernier ayant ete obtenu en analysant des donnees morphometriques. Les auteurs discutent egalement de l'utilite des AFLP pour la determination des relations phylogenetiques. Mots cles : Echinacea, AFLP, distance genetique, phylogenie.

Background/Objective Despite the relevance of pancreatic fat content in the development of metabolic diseases, its association with impaired glucose metabolism, diabetes, and other adipose tissue compartments remains unclear. Thus, we determined differences in pancreatic fat content by magnetic resonance imaging (MRI) between subjects with prediabetes, diabetes, and normal controls in a cohort from the general population. Methods Subjects without history of cardiovascular disease with established diabetes or prediabetes as well as normal controls were included and underwent whole-body MRI on a 3T scanner. Pancreatic fat content was quantified by measuring the proton-density fat fraction (PDFFpanc) using a 3D multi-echo GRE sequence (increment: 1.23 ms, 6 echoes) by placing ROIs in the pancreatic head, body, and tail by independent readers. In addition, hepatic fat content as well as abdominal subcutaneous and visceral adipose tissue (SAT and VAT) were measured by multi-echo GRE and 3D 2-point volume-interpolated DIXON MRI, respectively. Univariate and multivariate analyses were employed to determine associations. Results A total of 385 subjects were included in the analysis (median age: 57 years, 58.2% males), of them 53 were classified as subjects with diabetes, 95 as prediabetes, and 237 as controls (13.8%, 24.7%, and 61.6%; respectively). The median PDFFpanc was 5.2% [IQR 3.3±9.4], and significantly higher in subjects with prediabetes and diabetes as compared to controls (PDFFpanc: 6.2% [IQR: 3.5±12] vs. 8.6% [IQR: 4.3±17.5] vs. 4.9% [3.1±7.4], p0.12). While in univariate analysis BMI, PDFFhepatic, SAT and VAT were associated with PDFFpanc (all p

The digestive vacuole (DV) of Plasmodium falciparum, which is released into the bloodstream upon rupture of each parasitized red blood cell (RBC), was recently discovered to activate the alternative complement pathway. In the present work, we show that C3- and C5-convertases assembling on the parasitic organelle are able to provoke deposition of activated C3 and C5b-9 on non-infected bystander erythrocytes. Direct contact of DVs with cells is mandatory for the effect, and bystander complement deposition occurs focally, possibly at the sites of contact. Complement opsonization promotes protracted erythrophagocytosis by human macrophages, an effect that is magnified when ring-stage infected RBCs with reduced CD55 and CD59, or paroxysmal nocturnal hemoglobinuria (PNH)-RBCs lacking these complement inhibitors are employed as targets. Bystander attack can also directly induce lysis of PNH-RBCs. Direct evidence for complement activation and bystander attack mediated by DVs was obtained through immunohistochemical analyses of brain paraffin sections from autopsies of patients who had died of cerebral malaria. C3d and the assembled C5b-9 complex could be detected in all sections, colocalizing with and often extending locally beyond massive accumulations of DVs that were identified under polarized light. This is the first demonstration that a complement-activating particle can mediate opsonization of bystander cells to promote their antibody-independent phagocytosis. The phenomenon may act in concert with other pathomechanisms to promote the development of anemia in patients with severe malaria.

BACKGROUND 1,25 dihydroxyvitamin D3 (1,25D) and retinoids may play an important role in preventing progression of prostate cancer. METHODS We examined the ability of four novel 20-epi-vitamin D3 analogs (CB1093, KH1060, KH1266, and CB1267), either alone or in combination with 9-cis retinoic acid (RA) to inhibit colony growth of a human prostate cancer cell line, LNCaP, using soft agar as well as bone marrow stroma. Also, the effect of these analogs on the cell cycle and expression of Ki-67, p21waf−1, and p27kip1 in LNCaP cells was examined. RESULTS The analog CB1267 was the most potent, with 8 × 10−10 M of the analog inhibiting 50% colony growth (ED50) of LNCaP. 9-cis-RA also inhibited colony growth of LNCaP (ED50, 5 × 10−7 M). Combined, CB1267 and 9-cis-RA synergistically inhibited colony growth and significantly increased the number of LNCaP cells in G0/G1 phase. Cell cycle arrest was associated with increased levels of p21waf−1 and p27kip1 and decreased expression of Ki-67 protein. Pulse-exposure to this combination (5 × 10−8 M) irreversibly inhibited colony growth, both in soft agar and on normal human bone marrow stroma. CONCLUSIONS Combination of a new vitamin D3 analog (CB1267) and a retinoid (9-cis-RA) potently inhibited colony formation of LNCaP prostate cancer cells in vitro, suggesting further studies in animal models. This combination may afford an interesting therapeutic approach to low-burden prostate cancer. Prostate 40:141–149, 1999. © 1999 Wiley-Liss, Inc.

Using a novel combination of several experimental techniques for state selective ion and electron spectroscopy, we investigated the decay dynamics of "n1l1n2l2" (4 1 2 1 ≥ 3, l2 > 5) planetary states in strontium after five-laser excitation. As a result, at least two dominant and competing fragmentation mechanisms (autoionization and fluorescence–autoionization cascades) could be identified. Quantitative investigation of these mechanisms and their decomposition into different channels yield a fairly complete picture of the fragmentation dynamics in highly excited three–body Coulomb states.

The biologic effects of retinoids such as all-trans-retinoic acid (ATRA) and 9-cis-retinoic acid on proliferation and differentiation of hematopoietic cells are mediated by binding and activating two distinct families of transcription factors: the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). The RARs require heterodimerization with RXRs; in addition, RXRs can form homodimers, which can bind to DNA response elements that are either distinct or the same as those bound by the RAR/RXR heterodimers. Therefore, the two retinoid pathways provide sequences that are specific for effective DNA binding and activation of target genes. We have developed several series of novel synthetic retinoids that selectively interact with RXR/RXR homodimers and RAR/RXR heterodimers. We show here that SR11236 and SR11246, which are RXR-selective analogs, had little ability to inhibit clonal growth and induce differentiation of leukemic cells (HL- 60 cells and fresh acute myeloid leukemia cells). However, SR11249, SR11256, and LGD1069, which activated both RXR/RXR homodimers and RAR/RXR heterodimers, could inhibit clonal growth and induce differentiation of HL-60 cells as well as leukemic cells from patients, including those with acute promyelocytic leukemia (APL). This is similar to results observed with RAR/RXR-specific ligands. Interestingly, the combination of ATRA and either SR11249, SR11256, or LGD1069 showed synergistic effects in inducing differentiation of HL-60 cells. A retinoid (SR11238) with strong anti-AP-1 activity that did not activate the RARs and RXRs for gene transcription from the response element TREpal was inactive in our assay systems, suggesting that the antiproliferative effects of retinoids on leukemic cells is not mediated by inhibiting the AP-1 pathway. We conclude that the RAR/RXR pathway is more important than RXR/RXR pathway for differentiation and proliferation of acute myeloid leukemic cells, and certain retinoids or combination of retinoids with both RAR and RXR specificities may synergistically enhance the differentiation activity of ATRA, which may be relevant in several clinical situations.

Severe Plasmodium falciparum malaria evolves through the interplay among capillary sequestration of parasitized erythrocytes, deregulated inflammatory responses, and hemostasis dysfunction. After rupture, each parasitized erythrocyte releases not only infective merozoites, but also the digestive vacuole (DV), a membrane-bounded organelle containing the malaria pigment hemozoin. In the present study, we report that the intact organelle, but not isolated hemozoin, dually activates the alternative complement and the intrinsic clotting pathway. Procoagulant activity is destroyed by phospholipase C treatment, indicating a critical role of phospholipid head groups exposed at the DV surface. Intravenous injection of DVs caused alternative pathway complement consumption and provoked apathy and reduced nociceptive responses in rats. Ultrasonication destroyed complement-activating and procoagulant properties in vitro and rendered the DVs biologically inactive in vivo. Low-molecular-weight dextran sulfate blocked activation of both complement and coagulation and protected animals from the harmful effects of DV infusion. We surmise that in chronic malaria, complement activation by and opsonization of the DV may serve a useful function in directing hemozoin to phagocytic cells for safe disposal. However, when the waste disposal system of the host is overburdened, DVs may transform into a trigger of pathology and therefore represent a potential therapeutic target in severe malaria.

The General Well-Being Schedule (GWB) is a brief, reliable, and valid instrument used in population studies to assess psychological well-being, although its validity with African-Americans has yet to be established. This study evaluated the reliability, validity, and factor structure of the GWB in a sample of 599 overweight African-American women who participated in multicenter weight loss trial. The results of the factor analysis indicate that the GWB is primarily unidimensional and that the existence of the six hypothesized subscales was not supported. The GWB demonstrated evidence of concurrent and construct validity when examined in association with measures of self-concept, depression, and several health behaviors. The results of this study suggest that the GWB is a reliable and valid measure of psychological well-being in African-American women.

Bariatric surgery is being performed in increasing numbers in an era when reimbursements are being reduced. Academic health centers bear the responsibility for training surgeons to perform these operations yet must keep costs to a minimum and retain high quality. The UCLA Bariatric Surgery Program developed a clinical pathway for the pre- and postoperative management for gastric bypass patients to achieve these goals. Medical records for 182 consecutive gastric bypass patients were retrospectively reviewed before implementation of the pathway (Group I) during the fiscal year of 1998/1999. Data on average length of stay, average intensive care unit length of stay, average standard variable cost, percentage readmission rate, and percentage return to the operating room were collected. This information was compared with the data collected prospectively from 182 patients after implementation of the pathway in July of 1999 (Group II) during the fiscal year of 1999/2000. Hospital cost per admission was reduced by 40 per cent in Group II compared with Group I (P0.02). The average length of stay was reduced from 4.05 days in Group I to 3.17 days in Group II (P0.033). Overall readmission rate was decreased from 4.2 per cent in Group I to 3.2 per cent in Group II (P0.05). There were no differences in morbidities between both groups. The pathway reduced costs by reducing the hospital length of stay, intensive care unit utilization, and readmission rates. Quality was maintained as evidenced by a similar pattern of postoperative morbidities yet readmission rates were reduced. Our results indicate that implementation of a clinical pathway for bariatric surgery reduces cost and improves quality of care in an academic institution.

Pharmacokinetics of three morphine 3-esters-3-(2,2-dimethylvaleroyl) morphine (A), 3-(2-phenylbenzoyl) morphine (B), and 3-(2,2-diphenylpropionyl) morphine (C) was characterized after single oral administration in rabbits. Blood was sampled up to 24 h and cerebro-spinal fluid (CSF) was collected with the last blood sample. The concentration of the morphine 3-esters, morphine, morphine 3-glucuronide and morphine 6-glucuronide were determined in plasma and CSF using HPLC UV-detection. The morphine 3-esters were suggested to be a subject to marked presystemic elimination, since, in comparison to the administration of the un-esterified morphine, relatively low concentrations of morphine and morphine glucuronides were detected in plasma. The rate of disposition of morphine was dependent on the hydrolytic stability of the esters. The mean (+/- S.E.) plasma half-life of morphine was 0.9 +/- 0.2 h, 2.5 +/- 0.6 h and 3.5 +/- 3.5 h after the administration of A, B and C, respectively, compared to 0.9 +/- 0.2 h as estimated after the administration of non-esterified morphine. An analgesic effect will be achieved, since morphine was detected in CSF even 24 h after the application of the ester pro-drugs. It is concluded that esterification at the 3-position may be adapted to obtain sustained plasma levels of morphine.

Although dietary intake of tomatoes and tomato products containing lycopene has been reported to reduce the risk of prostate cancer, few studies have been done on the relationship between plasma lycopene and other carotenoids and prostate cancer. This case-control study was conducted to investigate the effects of plasma lycopene, other carotenoids, and retinol, as well as alpha- and gamma-tocopherols on the risk of prostate cancer. The study included 65 patients with prostate cancer and 132 cancer-free controls; all of them were interviewed using a standard epidemiological questionnaire at the Memorial Sloan-Kettering Cancer Center from 1993 to 1997. Plasma levels of carotenoids, retinol, and tocopherols were measured by high performance liquid chromatography. An unconditional logistic regression model was used in bivariate and multivariate analyses using Statistical Analysis System (SAS). After adjusting for age, race, years of education, daily caloric intake, pack-years of smoking, alcohol consumption, and family history of prostate cancer, significantly inverse associations with prostate cancer were observed with plasma concentrations of the following carotenoids: lycopene [odds ratio (OR), 0.17; 95% confidence interval (CI), 0.04-0.78; P for trend, 0.0052] and zeaxanthin (OR, 0.22; 95% CI, 0.06-0.83; P for trend, 0.0028) when comparing highest with lowest quartiles. Borderline associations were found for lutein (OR, 0.30; 95% CI, 0.09-1.03; P for trend, 0.0064) and beta-cryptoxanthin (OR, 0.31; 95% CI, 0.08-1.24; P for trend, 0.0666). No obvious associations were found for alpha- and beta-carotenes, retinol, and alpha- and gamma-tocopherols. Our study confirmed the inverse associations between lycopene, other carotenoids such as zeaxanthin, lutein, and beta-cryptoxanthin, and prostate cancer. This study provides justification for further research on the associations between lycopene and other antioxidants and the risk of prostate cancer.

A simple and sensitive method for the polarographic determination of praziquantel (1) after derivatization using Vilsmeier formylation is described. The polarographically active compound obtained by this procedure has been separated, identified and prepared using N,N-dimethylformamide and phosphorus oxychloride.

We studied lipolytic activities in vivo in golden mantle ground squirrels during pre-hibernation and hibernation using microdialysis technique. Microdialysis probes were inserted into the abdominal subcutaneous adipose tissues. Baseline lipolysis were assessed by measuring glycerol concentration. Epinephrine-stimulated lipolysis was also examined. Eight squirrels (four male, four female) were studied in each of the two stages. Basal glycerol concentrations were lower in the hibernating state than in the pre-hibernation state in male squirrels (P

1,25 dihydroxyvitamin D3 (1,25D) and retinoids may play an important role in preventing progression of prostate cancer.We examined the ability of four novel 20-epi-vitamin D3 analogs (CB1093, KH1060, KH1266, and CB1267), either alone or in combination with 9-cis retinoic acid (RA) to inhibit colony growth of a human prostate cancer cell line, LNCaP, using soft agar as well as bone marrow stroma. Also, the effect of these analogs on the cell cycle and expression of Ki-67, p21(waf-1), and p27(kip1) in LNCaP cells was examined.The analog CB1267 was the most potent, with 8 x 10(-10) M of the analog inhibiting 50% colony growth (ED50) of LNCaP. 9-cis-RA also inhibited colony growth of LNCaP (ED50, 5 x 10(-7) M). Combined, CB1267 and 9-cis-RA synergistically inhibited colony growth and significantly increased the number of LNCaP cells in G0/G1 phase. Cell cycle arrest was associated with increased levels of p21(waf-1) and p27(kip1) and decreased expression of Ki-67 protein. Pulse-exposure to this combination (5 x 10(-8) M) irreversibly inhibited colony growth, both in soft agar and on normal human bone marrow stroma.Combination of a new vitamin D3 analog (CB1267) and a retinoid (9-cis-RA) potently inhibited colony formation of LNCaP prostate cancer cells in vitro, suggesting further studies in animal models. This combination may afford an interesting therapeutic approach to low-burden prostate cancer.

Management of prostate cancer that has spread outside of the prostate capsule is a difficult problem. Innovative, non-toxic approaches to the disease are required. New, relatively non-toxic vitamin D3 analogs have recently been synthesized. We report that several of these compounds have marked antiproliferative effects on prostate cells.The clonal antiproliferative activity of five novel analogs of 1,25 dihydroxyvitamin D3 [1,25(OH)2D3, (cmpd C)] as well as 1,25(OH)2D3 itself was tested on three human prostate cancer cell lines (PC-3, LNCaP, and DU-145). The analogs were 20-epi-22oxa-24a,26a,27a-tri-homo-1 alpha,25(OH)2D3 (code name: KH 1060); 24a26a27a-tri-homo-22,24-diene-1 alpha,25(OH)2D3 (code name: EB 1089); 1,25(OH)2-16ene-D3 (code name: HM); 1,25(OH)2-16ene-23yne-D3 (code name: V); 1,25(OH)2-20-epi-D3 (code name: MC 1288)].With the parent compound [1,25(OH)2D3], the effective dose that inhibited 50% clonogenic growth of PC-3 and LNCaP was 10(-8)M and 7 x 10(-9)M, respectively. For these prostate cancer cell lines, KH 1060 was the most potent analog by an order of 25- to 35-fold as compared to cmpd C. The second and third most potent analogs were HM and MC 1288. DU-145 was resistant to all the vitamin D3 analogs. The major side-effect of 1,25(OH)2D3 is the production of hypercalcemia. The relative inhibitory index (RII) was determined by comparing the antiproliferative activity of the analog to its ability to produce hypercalcemia in mice injected intraperitoneally every other day. The KH 1060 had the best RTI: 50- to 70-fold greater than 1,25(OH)2D3 for PC-3 and LNCaP, respectively.A trial of one or more of these innovative compounds should be considered for treatment of minimal residual disease of prostate cancer.

Background Real-time sonoelastography (SElasto) is used to examine tissue elasticity. Chronic inflammation and fibrosis of major salivary glands leads to reduced saliva production in patients with primary Sjogren’s syndrome (pSS). Objectives To investigate the value of SElasto to detect pSS patients with reduced saliva production Methods Prospective study on 38 pSS patients fulfilling the American-European consensus group criteria [mean age 58 years; 92% female; median duration of sicca symptoms 6 years, 65% histological sialadenitis] and 11 healthy controls. B-mode sonography and SElasto of parotid and submandibular glands was performed using a GE Logiq E9 ultrasound device. Parenchymal homogenicity, echogenicity, hypoechogenic areals, hyperechoic reflections and clearness of glandular borders were semiquantitatively scored (total score ranging from 0-48) (1). SElasto was used to examine the elasticity of glandular parenchyma and a semiquantitative rating was performed with 0=no, 1=up to 25%, 2=up to 50%, 3=up to 75% and 4=more than 75% hardened areas within the salivary gland. The total score ranged from 0 to 16. Clinical assessments were performed at the same day of sonographic evaluation and included: Saxon test, Schirmer test, Xerostomia inventory (XI), and the Ocular Surface Disease Index (OSDI). Statistical analysis was performed using SPSS program (v18.0) and the Mann-Whitney-U and Spearman rank correlation test were performed as appropriate. Interobserver variability of sonography was tested in 30% of pSS patients by intraclass correlation coefficient (ICC). Results pSS patients had higher B-mode scores [median 25 (range 2.0-44.0) vs. 2.0 (0-8.0), p pSS patients showed a median salivary flow rate of 1.69 g/2min (range 0.31-3.79), a median moisture on the filter paper (Schirmer test) of 4.0mm/5min (0-50.0), a median XI of 27.5 (8.0-43.9) and an median OSDI of 43.8 (0-77.1). In pSS patients, an inverse correlation was found between the result of the Saxon test and SElasto score (corr coeff -0.426, p=0.009), whereas B-mode ultrasound results were not associated with saliva production. Neither disease duration nor duration of sicca symptoms influenced ultrasound results. A good reproducibility of B-mode and SElasto results was found as indicated by an ICC of 0.926 (95%CI 0.565-0.983) and 0.934 (0.787-0.981), respectively. Conclusions Increased rigidity of major salivary glands as demonstrated by SElasto in patients with pSS correlates with the impairment of saliva secretion. References Hocevar A, Ambrozic A, Rozman B, Kveder T, Tomsic M. Ultrasonographic changes of major salivary glands in primary Sjogren’s syndrome. Diagnostic value of a novel scoring system. Rheumatology 2005;44:768-72. Disclosure of Interest None Declared

Background Sialoscintigraphy (Szinti) is used to investigate salivary gland function in patients with primary Sjogren’s syndrome (pSS). Real-time sonoelastography (SElasto) indicates tissue rigidity of salivary glands and correlates with an impaired saliva production. Objectives To investigate the value of SElasto and B-mode sonography to identify pSS patients with dysfunctional salivary glands. Methods Prospective study on 37 pSS patients fulfilling the American-European consensus group criteria [mean age 59 years; 92% female; median disease duration 3.1 years]. Szinti was conducted according to a routine protocol and semiquantitative scoring was performed (1): each gland was graded into 3= severe dysfunction, 2= moderate dysfunction, 1= mild dysfunction or 0= normal function. B-mode sonography and SElasto of parotid and submandibular glands was performed using a GE Logiq E9 ultrasound device. Parenchymal homogenicity, echogenicity, hypoechogenic areals, hyperechoic reflections and clearness of glandular borders were also semiquantitatively scored (total score 0-48) (2). SElasto was used to examine the elasticity of glandular parenchyma and a semiquantitative rating was performed with 0=no, 1=up to 25%, 2=up to 50%, 3=up to 75% and 4=more than 75% hardened areas within the salivary gland (total score 0-16). Interobserver variability of sonography and Szinti were tested in 30% of pSS patients. Results The mean Szinti score of pSS patients was 6.0 (±4.3). Loss function of 1,2 or 4 salivary glands was present in 5.3%, 17.5% and 19.3% of patients, respectively. B-mode (corr coeff 0.65, p coeff 0.39, p=0.02) correlated with the Szinti score. Patients with at least one dysfunctional salivary gland had higher B-mode [median 27.5 (range 10.0-44.0) vs. 12.0 (2.0-6.9) p A good reproducibility of B-mode and SElasto results was found as indicated by an ICC of 0.926 (95%CI 0.565-0.983) and 0.934 (0.787-0.981), respectively. Reproducibility of Szinti results was also good (kappa 0.871). Conclusions Structural changes and increased rigidity of major salivary glands as demonstrated by B-mode sonography and SElasto, respectively correlates closely withsalivary gland dysfunction in patients with pSS. References Shizukuishi K, Nagaoka S, Kinno Y, Saito M, Takahashi N, Kawamoto M et al. Scoring analysis of salivary gland scintigraphy in patients with Sjogren’s syndrome. Ann Nucl Med 2003;17:627-31. Shizukuishi K, Nagaoka S, Kinno Y, Saito M, Takahashi N, Kawamoto M et al. Ultrasonographic changes of major salivary glands in primary Sjogren’s syndrome. Diagnostic value of a novel scoring system. Rheumatology 2005;44:768-72 Disclosure of Interest None Declared

The biologic effects of retinoids such as all-trans-retinoic acid (ATRA) and 9-cis-retinoic acid on proliferation and differentiation of hematopoietic cells are mediated by binding and activating two distinct families of transcription factors: the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). The RARs require heterodimerization with RXRs; in addition, RXRs can form homodimers, which can bind to DNA response elements that are either distinct or the same as those bound by the RAR/RXR heterodimers. Therefore, the two retinoid pathways provide sequences that are specific for effective DNA binding and activation of target genes. We have developed several series of novel synthetic retinoids that selectively interact with RXR/RXR homodimers and RAR/RXR heterodimers. We show here that SR11236 and SR11246, which are RXR-selective analogs, had little ability to inhibit clonal growth and induce differentiation of leukemic cells (HL-60 cells and fresh acute myeloid leukemia cells). However, SR11249, SR11256, and LGD1069, which activated both RXR/RXR homodimers and RAR/RXR heterodimers, could inhibit clonal growth and induce differentiation of HL-60 cells as well as leukemic cells from patients, including those with acute promyelocytic leukemia (APL). This is similar to results observed with RAR/RXR-specific ligands. Interestingly, the combination of ATRA and either SR11249, SR11256, or LGD1069 showed synergistic effects in inducing differentiation of HL-60 cells. A retinoid (SR11238) with strong anti-AP-1 activity that did not activate the RARs and RXRs for gene transcription from the response element TREpal was inactive in our assay systems, suggesting that the antiproliferative effects of retinoids on leukemic cells is not mediated by inhibiting the AP-1 pathway. We conclude that the RAR/RXR pathway is more important than RXR/RXR pathway for differentiation and proliferation of acute myeloid leukemic cells, and certain retinoids or combination of retinoids with both RAR and RXR specificities may synergistically enhance the differentiation activity of ATRA, which may be relevant in several clinical situations.

Breast cancer is a devastating disease. New approach in therapy are needed. In this chapter, we describe our efforts to identify novel vitamin D3 analogs which may have a potent antiproliferative effect on breast cancer without causing hypercalcemia.

Low fat, high fiber dietary interventions that decrease blood estrogen levels may reduce breast cancer risk. Asian women consuming their traditional low fat, high fiber diets have lower blood estrogen levels before and after menopause and lower rates of breast cancer compared with Western women. The current controlled feeding study of premenopausal women was designed to determine the effects of a very low fat (10% of calories) and high fiber (35-45 g/day) diet on blood estrogen levels and menstrual function.Twelve healthy premenopausal women with regular ovulatory cycles were followed for 3 months. Subjects consumed a diet providing 30% of their energy from fat and 15-25 g of dietary fiber per day for 1 month, and they consumed a very low fat, high fiber and libitum diet providing 10% of their energy from fat and 25-35 g of dietary fiber per day for 2 months.At the end of the second month of the very low fat, high fiber diet, there was a significant reduction in serum estrone and estradiol levels during the early follicular and late luteal phases. There were no significant changes observed in serum estrone sulfate, sex hormone binding globulin, or progesterone. Despite a significant decrease in serum estradiol and estrone levels after 2 months of a very low fat, high fiber diet, there was no interference with ovulation or the magnitude of the mid-cycle leuteinizing hormone surge. Small changes in menstrual cycle length of up to 3 days were not ruled out due to the small sample size of the study.A very low fat, high fiber diet in healthy premenopausal women can reduce estradiol and estrone levels without affecting ovulation, thereby providing a rationale for the prevention of breast cancer through a very low fat, high fiber diet.

2-Digitoxigenone amidinohydrazone (1), a compound with known digitalis-like activity, and Schiff bases 2-11 of 3-digitoxigenone were synthesized and tested pharmacologically in order to further determine possible structural requirements at the 3-position of digitalis compounds. The inotropic activity was screened using guinea-pig atria, and the interaction with the digitalis receptor was further examined using [3H]ouabain binding to cardiac membranes from guinea pig, rat, pit and man. All compounds revealed activities intermediate between 3-digitoxigenone and ouabain, and the potency of the derivatives covered approximately one order of magnitude. The absolute potency varied among species, but the rank order of potency was rather similar, yielding good correlations between species. This indicates no pronounced preference of these compounds for a particular (Na+/K+)-ATPase isoform of any of the species studied.

We have studied the in vitro biological activities and mechanism of action of 1,25-dihydroxyvitamin D3 (1,25D3) and four potent 1,25D3 analogues [20-epi-22oxa-24a,26a,27a-tri-homo-1,25(OH)2D3 (KH 1060); 20-epi-1,25(OH)2D3; 1,25(OH)2-16ene-D3; and 1,25(OH)2-16ene-23yne-D3] on proliferation and differentiation of estrogen receptor-negative (MDA-MB-436, BT-20, SK-BR-3, and MDA-MB-231), estrogen receptor-weakly positive (BT474), and estrogen receptor-positive (MCF-7) breast cancer cell lines. Dose-response studies showed that KH 1060 was the most potent analogue, because it was able to induce differentiation in all seven breast cancer cell lines (measured by lipid staining) and to suppress more than 50% clonal proliferation (ED50) at 10(-10) M in all cell lines, except MDA-MB-436 and BT-20. To explore how these compounds mediated antiproliferative actions, their effects on the cell cycle, on expression of bcl-2 and p53, and on apoptosis were assessed. Five of six cell lines have a mutant p53 gene, whereas MCF-7 has wild-type p53. Immunohistochemical staining showed that the p53 protein was predominantly localized in the nucleus in each of the breast cancer cell lines except for MCF-7, which expressed the protein predominantly in the cytoplasm. After incubation with KH 1060 (3 days; 10(-7) M), expression of bcl-2 protein as determined by immunohistochemical localization was markedly decreased in BT-474, MCF-7, and MDA-MB-231; these same cells were profoundly inhibited in their clonal proliferation and arrested in the G0/G1 phase of the cell cycle when cultured with KH 1060. In contrast, BT-20 and MDA-MB-436 cells that were refractory to the antiproliferative effect of KH 1060 (ED5010(-6) M) had no down-regulation of their bcl-2 expression and no cell cycle changes after exposure to KH 1060. MCF-7 showed morphological changes and DNA fragmentation, indicative of apoptosis after 48 h incubation with KH 1060 (10(-6) M), during which time p53 protein accumulated in the nucleus and decreased in the cytoplasm. In contrast, no apoptosis was detected in three other breast lines (MDA-MB-231, SK-BR-3, and BT-474) that had a mutated p53. In conclusion, the data indicate that KH 1060 is an extremely potent 1,25D3 analogue inducing differentiation of all six breast cancer lines and potently inhibiting clonal growth of four of them with concomitant decreased bcl-2 and cell cycle arrest at G0/G1.(ABSTRACT TRUNCATED AT 400 WORDS)