2B-Alert Web: An Open-Access Tool for Predicting the Effects of Sleep/Wake Schedules and Caffeine Consumption on Neurobehavioral Performance.

STUDY OBJECTIVES: Computational tools that predict the effects of daily sleep/wake amounts on neurobehavioral performance are critical components of fatigue management systems, allowing for the identification of periods during which individuals are at increased risk for performance errors. However, none of the existing computational tools is publicly available, and the commercially available tools do not account for the beneficial effects of caffeine on performance, limiting their practical utility. Here, we introduce 2B-Alert Web, an open-access tool for predicting neurobehavioral performance, which accounts for the effects of sleep/wake schedules, time of day, and caffeine consumption, while incorporating the latest scientific findings in sleep restriction, sleep extension, and recovery sleep. METHODS: We combined our validated Unified Model of Performance and our validated caffeine model to form a single, integrated modeling framework instantiated as a Web-enabled tool. 2B-Alert Web allows users to input daily sleep/wake schedules and caffeine consumption (dosage and time) to obtain group-average predictions of neurobehavioral performance based on psychomotor vigilance tasks. 2B-Alert Web is accessible at: https://2b-alert-web.bhsai.org. RESULTS: The 2B-Alert Web tool allows users to obtain predictions for mean response time, mean reciprocal response time, and number of lapses. The graphing tool allows for simultaneous display of up to seven different sleep/wake and caffeine schedules. The schedules and corresponding predicted outputs can be saved as a Microsoft Excel file; the corresponding plots can be saved as an image file. The schedules and predictions are erased when the user logs off, thereby maintaining privacy and confidentiality. CONCLUSIONS: The publicly accessible 2B-Alert Web tool is available for operators, schedulers, and neurobehavioral scientists as well as the general public to determine the impact of any given sleep/wake schedule, caffeine consumption, and time of day on performance of a group of individuals. This evidence-based tool can be used as a decision aid to design effective work schedules, guide the design of future sleep restriction and caffeine studies, and increase public awareness of the effects of sleep amounts, time of day, and caffeine on alertness.

The effect of metronomic versus standard chemotherapy on the regulatory to effector T-cell equilibrium in cancer patients.

BACKGROUND: The host's immune system is crucially involved in cancer development and progression. The ratio of regulatory to effector T-cells, as well as the interplay of T-cells with therapeutic agents, impact on cancer prognosis. The current study aimed to comparatively investigate the effect of metronomic and standard chemotherapy on the number and functionality of peripheral regulatory and effector T-cells in cancer patients. METHODS: CD4+CD25+ regulatory and CD4+CD25- effector T-cells were purified from the peripheral blood of 36 cancer patients and co-cultured in the presence of a polyclonal stimulus. The proliferative capacity and frequency of CD4+CD25+/CD4+CD25- T-cells were analysed before and during various chemotherapeutic regimes, by ELISA and flow cytometry, respectively. RESULTS: Chemotherapy shifted immune responses in favour of regulatory T-cells. The relative ratio of regulatory to effector T-cells increased, and the T-cell-mediated suppressive activity of regulatory on effector T-cells was augmented. This effect was more profound in metronomic than in standard chemotherapeutic approaches. Moreover, an association between the chemotherapy strategy followed and the mode of action of specific drugs (anti-mitotic, anti-DNA) was revealed. CONCLUSIONS: In comparison to standard chemotherapeutic strategies, metronomic approaches, though more patient-friendly, result in a significantly more prominent expansion of regulatory T-cells that aggravate the regulatory to effector T-cell imbalance. Our findings impact on the modulation of chemotherapy-treated patients' anti-tumor immunity and, thus, may be proven useful for selecting the most advantageous drug-delivery strategy, particularly when immunotherapeutics are eventually to be applied.

Thyroid hormones are positively associated with insulin resistance early in the development of type 2 diabetes.

Thyroid hormones have generally been found normal in diabetic patients. The question of whether variation within the euthyroid range influences insulin sensitivity in type 2 diabetes remains to be established. To investigate this, a meal was given to four groups: 17 healthy volunteers (controls), 22 first-degree relatives of type 2 diabetic subjects (relatives), 15 subjects with impaired glucose tolerance (IGT), and 24 subjects with overt type 2 diabetes (DM). Blood was drawn for 360 min for measurements of glucose and insulin. Plasma-free-T4(FT4) and plasma-free-T3(FT3) levels were measured. Fasting and postprandial insulin resistance was assessed by HOMA-IR and ISI indices, respectively. FT4 levels were found to be lower in controls (13.73 ± 0.48 pmol/l) than relatives, IGT, and DM (15.33 ± 0.52, 16.13 ± 0.65, and 17.7 ± 0.85 pmol/l, respectively, P = 0.007). FT3 levels were lower in controls (3.68 ± 0.09 pmol/l) than in relatives, IGT, and DM (4.35 ± 0.1, 4.8 ± 0.067, and 4.87 ± 0.11 pmol/l, respectively, P = 0.001). HOMA-IR was positively associated with FT4 and FT3 levels (ß-co-efficient = 1.876 ± 0.476, P = 0.001; and 0.406 ± 0.090, P = 0.001, respectively). ISI was negatively associated with FT4 and FT3 levels (ß-co-efficient = -0.051 ± 0.009, P = 0.001 and -0.009 ± 0.002, P = 0.001, respectively). In conclusion, increases of thyroid hormone levels within the normal range associate positively with insulin resistance. These data suggest that thyroid hormones may be part of the pathogenetic mechanism to explain metabolic derangement early in the development of type 2 diabetes.

Subcutaneous glucose monitoring with GlucoDay: comparison of the results to those obtained with the endocrine artificial pancreas.

OBJECTIVE: This study was undertaken to assess the accuracy of GlucoDay- a portable detector of subcutaneous glucose--by comparing the results to those obtained by Biostator an established and reliable method for continuous glucose measurement in whole blood. DESIGN: Subjects with type 1 diabetes (n:6), subjects with type 2 diabetes (n:6), and six healthy controls were studied for 24 hours; they consumed three main meals. The GlucoDay was connected to the subjects by inserting a microfibre probe into the periumbilical subcutaneous area, whilst the Biostator was inserted by a double-lumen catheter into an antecubital vein. A third catheter was inserted into a separate vein for blood withdrawal to measure glucose by the hexokinase method. RESULTS: The three methods (GlucoDay-Biostator-hexokinase) were equally accurate in measuring glucose levels (p = 0.233, Kruskall-Wallis test). The glucose measurements performed with GlucoDay and Biostator were significantly correlated with those performed with hexokinase (p < 0.001, r2 = 66.65% and p < 0.001, r2 = 64.4%, respectively, using simple regression analysis). CONCLUSIONS: Measurements of glucose fluctuations in the subcutaneous tissue with the GlucoDay were close to those in blood determined by the Biostator. GlucoDay is therefore a reliable method for continuous glucose monitoring and may prove useful for optimizating treatment in patients with type 1 or type 2 diabetes.

Rosuvastatin-induced thrombocytopenia.

Rosuvastatin, a statin indicated for patients with primary hypercholesterolemia, mixed dyslipidemia and familial hypercholesterolemia, is well tolerated by most patients. Its most common adverse effects are gastrointestinal derangement, muscle aches and hepatitis. One rare complication of statin treatment is severe thrombocytopenia. The case of a 65-year-old patient who developed severe thrombocytopenia while on rosuvastatin is presented, in addition to a review of the literature.

Insulin resistance in hyperthyroidism: the role of IL6 and TNF alpha.

OBJECTIVE: Although insulin resistance is a common finding in hyperthyroidism, the implicated mechanisms are obscure. The aim of this study was to investigate whether interleukin 6 (IL6) and tumour necrosis factor alpha (TNFalpha) are related to the development of insulin resistance in hyperthyroidism of nonautoimmune origin. DESIGN AND METHODS: A meal was given to ten hyperthyroid (HR) and ten euthyroid (EU) women. Plasma samples were taken for 360 min from the radial artery for measurements of glucose, insulin, and nonesterified fatty acids (NEFA). IL6 and TNFalpha were measured preprandially from the superficial epigastric vein and from the radial artery. RESULTS: i) In HR versus EU: (a) arterial glucose was similar (AUC(0-360) 2087+/-57 vs 2010+/-43 mM x min), but insulin was increased (AUC(0-360) 17 267+/-2447 vs 10 331+/-666 microU/ml x min, P=0.01), (b) homeostasis model assessment (HOMA) was increased (2.3+/-0.4 vs 1+/-0.1 kg/m(2), P=0.007), (c) arterial NEFA were increased (AUC(0-360) 136+/-18 vs 89+/-7 mmol/lxmin, P=0.03), (d) arterial IL6 (2+/-0.3 vs 0.9+/-0.1 pg/ml, P=0.0009) and TNFalpha (4.2+/-0.8 vs 1.5+/-0.2 pg/ml, P=0.003) were increased, and (e) IL6 production from the subcutaneous adipose tissue (AT) was increased (18+/-6 vs 5+/-1 pg/min per 100 ml tissue, P=0.04). ii) (a) Subcutaneous venous IL6 was positively associated with HOMA (beta-coefficient=1.7+/-0.7, P=0.049) and (b) although TNFalpha was not produced by the subcutaneous AT, arterial TNFalpha was positively associated with NEFA (AUC(0-360); beta-coefficient=0.045+/-0.01, P=0.005). CONCLUSIONS: In hyperthyroidism: i) glucose and lipid metabolism are resistant to insulin, ii) subcutaneous AT releases IL6, which could then act as an endocrine mediator of insulin resistance, iii) although there is no net secretion of TNFalpha by the subcutaneous AT, increased systemic TNFalpha levels may be related to the development of insulin resistance in lipolysis.

Methotrexate, paclitaxel, ifosfamide, and cisplatin in poor-risk nonseminomatous germ cell tumors.

OBJECTIVE: The efficacy and tolerability of M-TIP was evaluated as first-line treatment for patients with poor-risk germ cell tumors (GCT), according to International Germ Cell Cancer Collaborative Group (IGCCCG) criteria. PATIENTS AND METHODS: Thirty patients with poor-risk GCT were treated with M-TIP (methotrexate 250 mg/m(2) given as a 4-hour infusion with folinic acid rescue on day 1, paclitaxel 175 mg/m(2) given as a 3-hour infusion on day 1, followed by ifosfamide 1.2 g/m(2) given as a 2-hour infusion and cisplatin 20 mg/m(2) given as a 2-hour infusion on days 2 to 6) regimen for four cycles. RESULTS: Five (16.6%, 95% confidence interval [CI]: 2%-31%) patients achieved clinical complete response (cCR) with chemotherapy only, 15 (50%, 95% CI: 31-69%) patients pathologic complete response (pCR) (11 had necrosis/fibrosis and 4 had mature teratoma) and 3 (10%) patients surgical complete response (sCR) for an overall favorable response of 76.6%. Twenty-one patients are continuously disease-free at a median follow-up of 5.3 years (range 0.9-8.4+ years), resulting in a 5-year progression-free survival (PFS) rate of 66.6% (95% CI = 49%-85%) and a 5-year survival rate of 70% (95% CI = 53%-87%). Toxicity was generally mild except for myelotoxicity. Patients with febrile neutropenia were successfully treated with broad spectrum antibiotics and G-CSF support. Hematologic toxicity in this trial was ameliorated with the use of G-CSF. Neurotoxicity and nephrotoxicity were not a problem, since only 6.6% and 3.3% of patients developed sensory neuropathy and renal toxicity, respectively. CONCLUSION: M-TIP is a highly effective (high proportion of patients achieved long-term disease-free status, lack of relapses) and well tolerated regimen for first-line treatment of poor-risk GCT patients. These results have to be compared with the standard BEP chemotherapy or more intensive regimens in multicentre randomized trials.

Rates of glucose uptake in adipose tissue and muscle in vivo after a mixed meal in women with morbid obesity.

BACKGROUND AND AIMS: Although whole-body insulin resistance in obesity is established, information on insulin action in peripheral tissues, especially adipose tissue (AD), is limited. This study was undertaken in morbid obesity to investigate insulin action on glucose disposal in AD and muscle (M). SUBJECTS AND METHODS: A meal was given to 30 obese (age 34 +/- 1 yr, body mass index 47 +/- 1 kg/m(2)) and 10 nonobese women (age 39 +/- 4 yr, body mass index 23 +/- 1 kg/m(2)). Samples for glucose and insulin were taken for 360 min from veins draining the abdominal subcutaneous AD and forearm muscles and from the radial artery. Blood flow (BF) was measured in AD ((133)Xe) and M (plethysmography). RESULTS: The area under the curve divided by time (AUC(0-360 min)/360 min) in obese vs. nonobese was as follows: 1) arterial glucose was similar 6.04 +/- 0.2 vs. 5.67 +/- 0.1 mm), but insulin was increased (65.5 +/- 6.6 vs. 28.7 +/- 1.7 mU/liter, P = 0.0004); 2) BF was decreased (3 +/- 0.2 vs. 4.4 +/- 0.3 ml/min per 100 ml tissue in M, P = 0.002 and 1.8 +/- 0.1 vs. 3.7 +/- 0.3 ml/min per 100 ml tissue in AD, P < 0.0001); 3) glucose uptake was decreased (0.9 +/- 0.1 vs. 2.3 +/- 0.4 micromol/min per 100 ml tissue in M, P = 0.002 and 0.45 +/- 0.1 vs. 1.1 +/- 0.17 micromol/min per 100 ml tissue in AD, P = 0.01); 4) fractional glucose extraction was decreased in M (5 +/- 1 vs. 9 +/- 1%, P = 0.03), but was similar in AD (3 +/- 1 vs. 3.6 +/- 1.4%); 5) glucose uptake (per total fat mass) was increased (0.275 +/- 0.04 vs. 0.12 +/- 0.02 mmol/min, P = 0.027). CONCLUSION: In morbid obesity, the sensitivity of glucose metabolism to insulin is impaired in M, due to defects in insulin-stimulated glucose use and decreased BF, and in AD, at least in part, due to decreased BF. However, increased total fat mass provides a sink for the excess of glucose and compensates for insulin resistance.

Combination chemotherapy with docetaxel, vinorelbine and estramustine phosphate in metastatic androgen-resistant prostate cancer: a single institution experience.

UNLABELLED: The aim of this study was to evaluate the activity and toxicity of docetaxel, vinorelbine and oral estramustine in androgen-resistant prostate cancer (ARPC). PATIENTS AND METHODS: Fifty-two eligible patients were treated with docetaxel at 30 mg/m2 (day 1 and 8), vinorelbine at 20 mg/m2 (day 1 and 8), and oral estramustine of 280 mg p.o. (daily on days 1 to 7) every 3 weeks for 12 cycles. Patients with osseous metastases received zoledronic acid of 4 mg every 3 weeks. Low molecular weight heparin was administered on a prophylaxis basis to all patients. RESULTS: A prostate-specific antigen (PSA) response > or = 50% from baseline was obtained in 29 (56%; 95% confidence interval [CI], 42-70%) patients. Objective responses among the 25 patients with measurable disease were observed in 48% (95% CI, 27-69%), including 1 patient with complete response (CR) and 11 patients with partial response (PR). Patients with extraosseous only, skeletal only, and extraosseous and skeletal metastases showed different PSA responses (87% vs. 44% vs. 59%, respectively, p = 0.094). Furthermore, patients with soft tissue disease only showed insignificantly better PSA response than those with skeletal metastases (response rate: 87% vs. 50%, p = 0.064). The median progression-free survival was 7.6 months (95% CI, 6.7-8.4 months) and the median overall survival was 18.2 months (95% CI, 15.5-20.8 months). The only parameters which were found to have an impact on survival were the extent of disease and the baseline levels of PSA. Toxicity was generally mild except for myelotoxicity. Neutropenia grade 3/4 was recorded in 33% of patients and 6% experienced febrile neutropenia. Anemia and thrombocytopenia grade 3 or 4 were not a problem. Three patients (6%) developed grade 3 sensory neuropathy and 2 patients (4%) developed grade 3 fatigue. Edema grade 3 occurred in 1 (2%) patient and thromboembolism grade 3 occurred in 2 (4%) patients. CONCLUSION: The combination of docetaxel, vinorelbine and oral estramustine is a well-tolerated regimen with high biochemical and objective response rates in patients with ARPC.

Studies of insulin resistance in patients with clinical and subclinical hypothyroidism.

OBJECTIVE: Although clinical hypothyroidism (HO) is associated with insulin resistance, there is no information on insulin action in subclinical hypothyroidism (SHO). DESIGN AND METHODS: To investigate this, we assessed the sensitivity of glucose metabolism to insulin both in vivo (by an oral glucose tolerance test) and in vitro (by measuring insulin-stimulated rates of glucose transport in isolated monocytes with flow cytometry) in 21 euthyroid subjects (EU), 12 patients with HO, and 13 patients with SHO. RESULTS: All three groups had comparable plasma glucose levels, with the HO and SHO having higher plasma insulin than the EU (P<0.05). Homeostasis model assessment index was increased in HO (1.97+/-0.22) and SHO (1.99+/-0.13) versus EU (1.27+/-0.16, P<0.05), while Matsuda index was decreased in HO (3.89+/-0.36) and SHO (4.26+/-0.48) versus EU (7.76+/-0.87, P<0.001), suggesting insulin resistance in both fasting and post-glucose state. At 100 microU/ml insulin: i) GLUT4 levels on the monocyte plasma membrane were decreased in both HO (215+/-19 mean fluorescence intensity, MFI) and SHO (218+/-24 MFI) versus EU (270+/-25 MFI, P=0.03 and 0.04 respectively), and ii) glucose transport rates in monocytes from HO (481+/-30 MFI) and SHO (462+/-19 MFI) were decreased versus EU (571+/-15 MFI, P=0.04 and 0.004 respectively). CONCLUSIONS: In patients with HO and SHO: i) insulin resistance was comparable; ii) insulin-stimulated rates of glucose transport in isolated monocytes were decreased due to impaired translocation of GLUT4 glucose transporters on the plasma membrane; iii) these findings could justify the increased risk for insulin resistance-associated disorders, such as cardiovascular disease, observed in patients with HO or SHO.

Combination chemotherapy with carboplatin, paclitaxel and pegylated liposomal doxorubicin for advanced or recurrent carcinosarcoma of the uterus: clinical experience of a single institution.

OBJECTIVES: The purpose of this study was to evaluate the activity and toxicity of carboplatin, paclitaxel and pegylated liposomal doxorubicin combination in advanced or recurrent of the uterine carcinosarcoma. METHODS: Twenty-nine eligible patients with measurable disease were treated with carboplatin [area under the curve (AUC) 5], paclitaxel 175 mg/m(2) and pegylated liposomal doxorubicin 25 mg/m(2) every 3 weeks for 6-8 cycles. RESULTS: There were 10 complete responses (CRs) (34%) and 8 partial responses (PRs) (28%) for an overall response rate (RR) of 62% (95% confidence interval [CI], 43-81%). The median progression-free survival (PFS) was 8.2 months (95% CI, 4.1-12.2 months) and the median overall survival (OS) was 16.4 months (95% CI, 14.7-18.0 months). There was no statistically significant difference between histology and response to therapy. Patients with PS of 0 or 1 had a higher RR than those with worst PS. Toxicity was generally mild except for myelotoxicity. Neutropenia grade 3/4 was recorded in 52% of patients and 10% experienced febrile neutropenia. Anemia grade 3 or 4 developed in 27% of patients and thrombocytopenia grade 3 or 4 in 31% of patients. Three patients (10%) developed grade 3 sensory neuropathy and only 2 patients (8%) grade 3 palmar-plantar erythrodysesthesias. No treatment-related deaths were recorded in our series. CONCLUSION: The combination of carboplatin, paclitaxel and pegylated liposomal doxorubicin appears to have activity in advanced, persistent or recurrent endometrial carcinosarcoma with an acceptable toxicity profile.

Migration and retraction of endothelial and epithelial cells require PHI-1, a specific protein-phosphatase-1 inhibitor protein.

Cell migration and retraction are interrelated activities that are crucial for a range of physiological processes such as wound healing and vascular permeability. Immunostaining of brain sections for the specific inhibitor of type-1 protein Ser/Thr phosphatase called PHI-1 showed high expression levels in smooth muscle and especially in vascular endothelial cells. During migration of cultured human lung microvascular endothelial cells, endogenous PHI-1 was concentrated to the trailing edge of the cells. Knockdown of PHI-1 using small interfering RNAs reduced by 45% the rate of HeLa cell migration in a wound-healing assay. These cells exhibited an extremely elongated phenotype relative to controls and time-lapse movies revealed a defect in retraction of the trailing edge. Both HeLa and human vascular endothelial cells depleted of PHI-1 showed increased surface areas relative to controls during cell spreading in a replating assay. Analysis of sequential microscopic images demonstrated this was due to a significant decrease in the number of retraction events, whereas protrusive action was unaffected. The Ser/Thr phosphorylation of several signaling, cytoskeletal and focal-adhesion proteins was unchanged in PHI-1-depleted cells, so the target of PHI-1 inhibited protein-phosphatase 1 remains unidentified. Nonetheless, the results show that PHI-1 participates in regulatory events at the trailing edge of migrating cells and modulates retraction of endothelial and epithelial cells.

Juxtamembrane localization of the protein phosphatase-1 inhibitor protein PHI-1 in smooth muscle cells.

Protein phosphorylation regulates many fundamental processes and protein phosphatase-1 (PP1) is a major phosphatase that determines the levels of Ser/Thr phosphorylation. Regulatory subunits and inhibitor phosphoproteins control PP1 activity. PHI-1 is a member of a family of PP1 inhibitor phosphoproteins that was discovered based on sequence similarity to the known inhibitor CPI-17. To learn more about PHI-1 we determined the tissue distribution of PHI-1 in embryonic and adult tissues, and examined its cellular localization by immunohistochemistry. In the embryo PHI-1 appeared first in the heart at E10, and by E15 it was detected in multiple tissues. Expression in adult tissues was strikingly different, with PHI-1 detected primarily in smooth muscles in the intestine, blood vessels, and male and female genitourinary tracts. PHI-1 also was highly expressed in the endothelial layer of blood vessels. Both PHI-1 and CPI-17 are expressed predominantly in adult smooth muscles. Whereas CPI-17 staining was diffuse PHI-1 was concentrated along the cell membrane in distinct foci, detected by confocal and electron microscopy. The common tissue distribution but different cellular localization of PHI-1 and CPI-17 suggest distinctive physiological roles for these two PP1 inhibitors.

The methylated component of the Neurospora crassa genome.

Cytosine methylation is common, but not ubiquitous, in eukaryotes. Mammals and the fungus Neurospora crassa have about 2-3% of cytosines methylated. In mammals, methylation is almost exclusively in the under-represented CpG dinucleotides, and most CpGs are methylated whereas in Neurospora, methylation is not preferentially in CpG dinucleotides and the bulk of the genome is unmethylated. DNA methylation is essential in mammals but is dispensable in Neurospora, making this simple eukaryote a favoured organism in which to study methylation. Recent studies indicate that DNA methylation in Neurospora depends on one DNA methyltransferase, DIM-2 (ref. 6), directed by a histone H3 methyltransferase, DIM-5 (ref. 7), but little is known about its cellular and evolutionary functions. As only four methylated sequences have been reported previously in N. crassa, we used methyl-binding-domain agarose chromatography to isolate the methylated component of the genome. DNA sequence analysis shows that the methylated component of the genome consists almost exclusively of relics of transposons that were subject to repeat-induced point mutation--a genome defence system that mutates duplicated sequences.