Efficacy of alphas1-casein hydrolysate on stress-related symptoms in women

OBJECTIVE: To examine the effects of alpha (s1)-casein hydrolysate on females with stress-related symptoms. DESIGN: Double-blind, randomized, crossover, placebo-controlled trial. SETTING: The alpha (s1)-casein hydrolysate was manufactured by INGREDIA (Arras, France) and the placebo was manufactured by DIETAROMA (Bourg, France). Study was designed and performed at PROCLAIM (Rennes, France), and the statistical analyses were performed by D Desor (Nancy, France). SUBJECTS: A total of 63 female volunteers suffering from at least one disorder that may be related to stress such as anxiety, sleep problems and general fatigue. INTERVENTIONS: A total of 63 volunteers participated in a double-blind, randomized, crossover, placebo-controlled study. Subjects were randomly allocated to receive either tablets containing alpha (s1)-casein hydrolysate or placebo at the dose of 150 mg/day for 30 days. After a 3 weeks washout period, they were crossed over for a new 30-day period of tablets intake. The outcome measure was a questionnaire including 44 items of symptoms that may be related stress in which the severity of each sign was evaluated using a 10-degree scale. These measures were studied repeatedly at the day of 0, 15 and 30 after the start of each interventional period. RESULTS: The 30-day treatment by alpha (s1)-casein hydrolysate in females with stress-related symptoms reduced their symptoms, particularly in digestion (P<0.01), cardiovascular (P<0.05), intellectual (P<0.01), emotional (P<0.05) and social problems (P<0.05). CONCLUSION: This study showed that a 30-day ingestion of alpha (s1)-casein hydrolysate decreased the stress-related symptoms in females suggesting that this product may be used as an effective functional ingredient alleviating such symptoms. Sponsorship: This study was partially supported by the INGREDIA of France and Neurobiology Research Program from the Korea Ministry of Science and Technology (2004-01757) of Korea.     

Prevalence and natural history of monoclonal and polyclonal B-cell lymphocytosis in a residential adult population

BACKGROUND: Monoclonal B-cells can be detected in the peripheral blood of some adults without B-cell malignancies, a condition recently termed monoclonal B-cell lymphocytosis (MBL). The risk of individuals with MBL progressing to a B-cell malignancy is unknown. Polyclonal B-cell lymphocytosis (PCBL) has not been systematically studied in the general population. METHODS: We obtained lymphocyte subset counts on 1,926 residential adults aged 40-76 years in a series of environmental health studies between 1991 and 1994. We then conducted two follow-ups in 1997 and 2003 on consenting participants with B-cell lymphocytosis, which included nine participants with MBL. To ascertain the clinical implications of MBL, we reviewed medical records and death certificates. RESULTS: The overall prevalence of MBL was 0.57% (11/1,926): nine cases at baseline and two additional cases identified at follow-up. Two (19%) MBL cases subsequently developed a B-cell malignancy; MBL persisted in the remaining nine cases (81%). All PCBL cases where no clone emerged regressed to normal B-cell counts over the follow-up period. MBL was significantly more frequent in residents near a hazardous waste site than in the control populations (age-adjusted OR 6.2; 95%CI 1.1-36.2). CONCLUSION: MBL confers an elevated risk for developing a B-cell malignancy, although it occurs only in a minority of cases. PCBL is most often a transient state, but a monoclonal population can emerge and persist. Prospective studies are needed to distinguish stable from progressive forms of B-cell lymphocytosis and to clarify the etiologic role of environmental exposures.     

Ejection load changes in aortic stenosis. Observations made after balloon aortic valvuloplasty.

To investigate complementarity and competitiveness between the intrinsic and extrinsic components of the total left ventricular systolic load, hemodynamic data from 18 elderly subjects with severe aortic stenosis were analyzed before and after balloon dilation of the stenosed aortic valve. Multisensor micromanometric pressure measurements allowed calculation (simplified Bernoulli equation) of the ejection velocity and aortic input impedance spectra. Despite a 32% increase in the aortic valve area (from 0.56 +/- 0.04 to 0.74 +/- 0.05 cm2 [mean +/- SEM], p < 0.01), the peak left ventricular systolic pressure fell by only 12% (from 189 +/- 10 to 167 +/- 8 mm Hg, p < 0.01). This was accompanied by an increase in the impedance at the same cardiac output. In a subset of patients (n = 9) in whom the peak aortic systolic pressure rose after valvuloplasty (from 115 +/- 10 to 128 +/- 12 mm Hg, p < 0.01), a 40% increase in the aortic valve area was accompanied by a marked increase in the aortic input impedance. In this subset, the steady component of the aortic input impedance increased by 24% (from 960 +/- 96 to 1,188 +/- 134 dyne.sec/ml, p < 0.05), and the characteristic impedance increased by 25% (from 106 +/- 13 to 132 +/- 19 dyne.sec/ml, p < 0.05). Because of an increased aortic impedance acutely following the procedure, the total left ventricular systolic load after balloon dilation of the stenotic valve was only slightly decreased despite a significant increase in aortic valve area. This represents an example of complementarity and competitiveness between the intrinsic and extrinsic components of the total systolic ventricular load. It may explain why improvement in left ventricular performance may be modest acutely following balloon aortic valvuloplasty.     

Fabrication of a complete,removable dental prosthesis from a digital intraoral impression for a patient with an excessively tight reconstructed lip after oral cancer treatment: A clinical report

This clinical report describes the management of a patient who had an excessively tight reconstructed lip because of oral cancer surgery and postoperative radiotherapy. The presented technique used an intraoral scanner for a preliminary impression and computer-aided design and computer-aided manufacturing (CAD-CAM) technology for preliminary laboratory procedures. This digital impression technique may reduce patient discomfort.     

Antifungal drug itraconazole targets VDAC1 to modulate the AMPK/mTOR signaling axis in endothelial cells

Itraconazole, a clinically used antifungal drug, was found to possess potent antiangiogenic and anticancer activity that is unique among the azole antifungals. Previous mechanistic studies have shown that itraconazole inhibits the mechanistic target of rapamycin (mTOR) signaling pathway, which is known to be a critical regulator of endothelial cell function and angiogenesis. However, the molecular target of itraconazole that mediates this activity has remained unknown. Here we identify the major target of itraconazole in endothelial cells as the mitochondrial protein voltage-dependent anion channel 1 (VDAC1), which regulates mitochondrial metabolism by controlling the passage of ions and small metabolites through the outer mitochondrial membrane. VDAC1 knockdown profoundly inhibits mTOR activity and cell proliferation in human umbilical vein cells (HUVEC), uncovering a previously unknown connection between VDAC1 and mTOR. Inhibition of VDAC1 by itraconazole disrupts mitochondrial metabolism, leading to an increase in the cellular AMP:ATP ratio and activation of the AMP-activated protein kinase (AMPK), an upstream regulator of mTOR. VDAC1-knockout cells are resistant to AMPK activation and mTOR inhibition by itraconazole, demonstrating that VDAC1 is the mediator of this activity. In addition, another known VDAC-targeting compound, erastin, also activates AMPK and inhibits mTOR and proliferation in HUVEC. VDAC1 thus represents a novel upstream regulator of mTOR signaling in endothelial cells and a promising target for the development of angiogenesis inhibitors.Angiogenesis, or the formation of new blood vessels from preexisting vasculature, is a critical process both in normal development and in the pathogenesis of a myriad of diseases. In particular, it has long been recognized that angiogenesis is required for tumor growth and metastasis and that growing tumors can promote angiogenesis by secreting proangiogenic factors, such as VEGF, basic FGF, EGF, and others (1, 2). These proangiogenic factors stimulate the proliferation, migration, and differentiation of the endothelial cells that make up the inner layer of all blood vessels, causing them to form new vessels that grow toward the source of these factors. This process, termed “tumor angiogenesis,” allows the tumor to keep up with an increasing demand for oxygen and nutrients as it grows, eliminate accumulating waste products, and shed cancerous cells into circulation leading to metastasis. Without angiogenesis, a tumor cannot grow larger than about 1–2 mm in diameter, the largest size at which nutrients can permeate by diffusion alone, and thus is rendered essentially harmless to the host (3). Inhibition of angiogenesis is emerging as a useful strategy for treating cancer. The discovery and development of angiogenesis inhibitors as therapeutics for cancer has culminated in the approval by the Food and Drug Administration (FDA) of a few antiangiogenic drugs. Bevacizumab (Avastin), a monoclonal antibody targeting VEGF, gained FDA approval for the treatment of metastatic colorectal cancer (4, 5). Pegaptanib (Macugen), a polynucleotide-based aptamer targeting VEGF (6), also has been approved by the FDA for the treatment of age-dependent macular degeneration. More recently, several kinase inhibitors, including sorafenib, sunitinib, pazopanib, and everolimus, that have a major, albeit nonspecific, effect on angiogenesis also have entered the clinic (7).Drug discovery and development is a time-consuming and costly process. The discovery and development of antiangiogenic drugs is no exception. To accelerate the process, we began a new initiative to collect known drugs and assemble them into what is now known as the “Johns Hopkins Drug Library” (JHDL). Screening of JHDL using an endothelial cell proliferation assay led to the identification of a number of hits. Among the most interesting hits is the antifungal drug itraconazole (8). Itraconazole potently inhibits endothelial cell proliferation with an IC₅₀ value (ca. 200 nM) that is significantly below its peak plasma levels (>2 µM) (9), suggesting that it is likely to have antiangiogenic activity under existing drug-administration regimens. It also displays high cell-type selectivity, being most potent against primary human endothelial cells in comparison with human foreskin fibroblasts and most human cancer cell lines. Moreover, it is more than 30-fold more potent than other members of the azole family of antifungal drugs, including ketoconazole and terconazole (8).Itraconazole was developed originally as an antifungal drug and has been used clinically for more than 30 years with a well-established safety record. Upon validation of its antiangiogenic and antitumor activity in a number of models both in vitro and in vivo (8, 10, 11), it entered multiple phase 2 clinical trials for treating cancer. To date, the pilot trials in nonsmall cell lung cancer, prostate cancer, and basal cell carcinoma have been completed; itraconazole has been shown to increase the progression-free and overall survival of patients taking the drug (12–14). Additionally, retrospective studies assessing the outcomes of patients with ovarian cancer and recurrent triple-negative breast cancer taking itraconazole have revealed significant increases in overall survival (15–17), which are likely to be attributable, at least in part, to the antiangiogenic activity of itraconazole. Thus, itraconazole has great potential for becoming a new drug to treat cancer and other angiogenesis-dependent diseases such as macular degeneration and diabetic retinopathy.Despite these promising clinical results, the mechanism by which itraconazole inhibits angiogenesis has remained largely unknown. Like other azole-containing antifungal drugs, itraconazole exerts its antifungal activity by inhibiting the CYP450 enzyme lanosterol 14-α demethylase (14DM), which is required for synthesis of the lipid ergosterol that maintains cell wall integrity in these organisms. However, the potency of itraconazole against human 14DM has been shown to be greatly reduced compared with the fungal enzyme (18, 19). Moreover, itraconazole is unique among this class of antifungal drugs in its antiangiogenic activity, including those that are more potent inhibitors of human 14DM (8). Taken together, these lines of evidence strongly suggest that 14DM inhibition cannot explain the antiangiogenic activity of itraconazole.In an attempt to identify the cellular pathways affected by itraconazole, we found that the mechanistic target of rapamycin (mTOR) signaling pathway, which regulates cell proliferation and is known to be required for angiogenesis, is potently inhibited by itraconazole at concentrations similar to those required for proliferation inhibition (20). This mTOR inhibitory activity also is more potent in endothelial cells, both primary and immortalized, than in other common cell lines such as HEK293T and HeLa cells. We found that inhibition of mTOR was mediated only in part by the inhibition of cholesterol trafficking through the endolysosome, leaving unanswered the question of what is the direct molecular target of itraconazole.In the present study, we designed and synthesized a photoaffinity probe of itraconazole to isolate and identify its binding proteins from live endothelial cells. Importantly, the probe retained full activity in endothelial cells, indicating that it binds to the same target proteins as itraconazole itself. Using a combination of affinity pulldown and mass spectrometry, we identified voltage-dependent anion channel 1 (VDAC1) as a primary binding protein of itraconazole. We demonstrated that itraconazole not only binds directly to VDAC1 but also interferes with its primary cellular function of regulating mitochondrial metabolism, causing a drop in cellular energy levels that triggers the energy-sensing protein AMP-activated protein kinase (AMPK). Subsequently, AMPK down-regulates mTOR activity through direct phosphorylation of the regulatory-associated protein of mTOR (raptor), ultimately leading to inhibition of endothelial cell proliferation.     

Pulmonary nodule size evaluation with chest tomosynthesis and CT: a phantom study

Objective:

We compared digital tomosynthesis (TOMO) and chest CT in terms of assessing the sizes of nodules located in zones where evaluation by simple radiography is limited.

Methods:

A total of 48 images comprising phantom nodules of four sizes in six different locations were used. Nodule size measurement errors for measurements using TOMO and CT images compared with the actual size from each observer were calculated. The inter- and intraobserver repeatability of the measured values and the agreement between the two techniques were assessed using the method described by Bland and Altman.

Results:

The mean measurement errors for all of the nodules and four observers were −0.84 mm [standard deviation (SD), 0.60 mm] on TOMO and −0.18 mm (SD, 0.71 mm) on CT images. The mean measurement errors for the different observers ranged from −1.11 to −0.55 mm for TOMO and from −0.39 to 0.08 mm for CT. Assessing the agreement between nodule size measurements using TOMO and CT resulted in mean measurement errors of −0.65 mm, with a 95% limit of agreement of −2.53 to 1.22 mm for comparison of TOMO with CT.

Conclusion:

Our results suggest that nodule sizes obtained using TOMO and chest CT are comparable, even for nodules located in areas where the size measurement is limited on simple radiography.

Advances in knowledge:

TOMO and CT can be used interchangeably, even for nodules located in a blind area on simple radiography.Solitary lung nodule detection has increased owing to the widespread use of CT imaging. Nevertheless, the most commonly used routine examination for lung nodules continues to be chest radiography, because it uses low radiation doses, is economical and is easy to use. Because chest radiographic images are two-dimensional projections of three-dimensional structures, early lung cancer detection on chest radiographs is often challenging. The projection of pulmonary vessels, bones and part of the mediastinum on lung fields often partially or completely obscures the pulmonary nodules, resulting in failure by the radiologist to detect lung nodules.^1,2Digital tomosynthesis (TOMO) has recently been applied to chest imaging for the detection of subtle nodules on simple radiography, with promising results.^3,4 It has been introduced as a modality with the potential to provide images similar to CT but at a comparably reduced cost and radiation exposure.⁴ James et al⁵ reported that 74% of lung nodules ≥4 mm in diameter that can be identified on CT can also be detected using TOMO. Vikgren et al⁶ also reported that 92% of nodules ≥4 mm in diameter are detectable using TOMO. In 2012, Johnsson et al⁷ compared the ability of TOMO and CT to detect nodule size in 20 patients and found that both methods could be used interchangeably for these measurements. This result calls for caution, however, because the limit of agreement (LOA) between the modalities is wider than for the intraobserver variability of each modality.Based on these studies, we hypothesized that TOMO is comparable to CT imaging for the detection of nodules located in areas where size measurement is limited using simple chest radiography because of overlapping structures. The purpose of this study was to assess the size determination of nodules located in these zones by TOMO and chest CT.