PMID- 18285398 TI - Adenosine modulates oesophageal sensorimotor function in humans. AB - BACKGROUND AND AIMS: Adenosine mediates somatic and visceral pain, but its effects on gut visceral nociception are unknown. The aim of the present study was to test the hypothesis that adenosine alters oesophageal sensorimotor function. METHODS: In a double-blind, randomised, placebo-controlled study, 14 healthy volunteers (M/F = 4/10) received either intravenous adenosine 100 microg/kg/min or placebo infusion. Prior to and during infusion, all subjects underwent stepwise graded oesophageal balloon distensions using impedance planimetry. Sensory responses and biomechanical properties were assessed and compared. RESULTS: Adenosine significantly lowered thresholds for first perception (median (25th-75th), cm H(2)O; 10 (10-20) vs 30 (20-30), p = 0.007), discomfort (40 (30 40) vs 50 (50-60), p = 0.011) and pain (50 (40-60) vs 70 (60-70), p = 0.007) when compared with placebo. Also, the median threshold pressures required to induce first perception (p = 0.017), discomfort (p = 0.024) and pain (p = 0.026) were lower when compared with baseline. The cross-sectional area of the oesophagus increased (p = 0.032), and the circumferential wall tension/strain relationship shifted to the left (the wall became stiffer) (p = 0.043) after adenosine, when compared with baseline or placebo. CONCLUSIONS: Adenosine can induce visceral hyperalgesia and decrease oesophageal distensibility in humans. These evoked sensorimotor changes are similar to those described in patients with functional oesophageal (non-cardiac) chest pain. Thus, adenosine modulates oesophageal sensorimotor function and may play a role in the pathogenesis of functional chest pain. PMID- 18285399 TI - Inherited intracellular ecosystem: symbiotic bacteria share bacteriocytes in whiteflies. AB - Symbiotic relationships with bacteria are common within the Arthropoda, with interactions that substantially influence the biology of both partners. The symbionts' spatial distribution is essential for understanding key aspects of this relationship, such as bacterial transmission, phenotype, and dynamics. In this study, fluorescence in situ hybridization was used to localize five secondary symbionts from various populations and biotypes of the sweet potato whitefly Bemisia tabaci: Hamiltonella, Arsenophonus, Cardinium, Wolbachia, and Rickettsia. All five symbionts were found to be located with the primary symbiont Portiera inside the bacteriocytes--cells specifically modified to house bacteria- but within these cells, they occupied various niches. The intrabacteriocyte distribution pattern of Rickettsia differed from what has been described previously. Cardinium and Wolbachia were found in other host tissues as well. Because all symbionts share the same cell, bacteriocytes in B. tabaci represent a unique intracellular ecosystem. This phenomenon may be a result of the direct enclosure of the bacteriocyte in the egg during oogenesis, providing a useful mechanism for efficient vertical transmission by "hitching a ride" with Portiera. On the other hand, cohabitation in the same cell provides ample opportunities for interactions among symbionts that can either facilitate (cooperation) or limit (warfare) symbiotic existence. PMID- 18285401 TI - Nitric oxide protects mast cells from activation-induced cell death: the role of the phosphatidylinositol-3 kinase-Akt-endothelial nitric oxide synthase pathway. AB - NO is known to suppress mast cell activation, but the role of NO in mast cell survival is unclear. Ligation of the high-affinity receptor for IgE (FcepsilonRI) resulted in NO production in mast cells within minutes. This NO production was largely dependent on NO synthase (NOS) activity and extracellular Ca(2+). The NO production required an aggregation of FcepsilonRI and was accompanied by increased phosphorylation of endothelial NOS (eNOS) at Ser1177 and Akt at Ser473. The phosphorylation of eNOS and Akt and the production of NO were abolished by the PI-3K inhibitor wortmannin. Although thapsigargin (TG) induced NO production as well, this response occurred with a considerable lag time (>10 min) and was independent of FcepsilonRI aggregation and PI-3K and NOS activity. Mast cells underwent apoptosis in response to TG but not upon FcepsilonRI ligation. However, when the NOS-dependent NO production was blocked, FcepsilonRI ligation caused sizable apoptosis, substantial mitochondrial cytochrome c release, caspase-3/7 activation, and collapse of the mitochondrial membrane potential, all of which were inhibited by the caspase-3 inhibitor z-Asp-Glu-Val-Asp-fluoromethylketone. The data suggest that the NO produced by the PI-3K-Akt-eNOS pathway is involved in protecting mast cells from cell death. PMID- 18285400 TI - Engineering insulin-like growth factor-1 for local delivery. AB - Insulin-like growth factor-1 (IGF-1) is a small protein that promotes cell survival and growth, often acting over long distances. Although for decades IGF-1 has been considered to have therapeutic potential, systemic side effects of IGF-1 are significant, and local delivery of IGF-1 for tissue repair has been a long standing challenge. In this study, we designed and purified a novel protein, heparin-binding IGF-1 (Xp-HB-IGF-1), which is a fusion protein of native IGF-1 with the heparin-binding domain of heparin-binding epidermal growth factor-like growth factor. Xp-HB-IGF-1 bound selectively to heparin as well as the cell surfaces of 3T3 fibroblasts, neonatal cardiac myocytes and differentiating ES cells. Xp-HB-IGF-1 activated the IGF-1 receptor and Akt with identical kinetics and dose response, indicating no compromise of biological activity due to the heparin-binding domain. Because cartilage is a proteoglycan-rich environment and IGF-1 is a known stimulus for chondrocyte biosynthesis, we then studied the effectiveness of Xp-HB-IGF-1 in cartilage. Xp-HB-IGF-1 was selectively retained by cartilage explants and led to sustained chondrocyte proteoglycan biosynthesis compared to IGF-1. These data show that the strategy of engineering a "long distance" growth factor like IGF-1 for local delivery may be useful for tissue repair and minimizing systemic effects. PMID- 18285402 TI - The secretory leukocyte protease inhibitor (SLPI) and the secondary granule protein lactoferrin are synthesized in myelocytes, colocalize in subcellular fractions of neutrophils, and are coreleased by activated neutrophils. AB - The secretory leukocyte protease inhibitor (SLPI) re-establishes homeostasis at sites of infection by virtue of its ability to exert antimicrobial activity, to suppress LPS-induced cellular immune responses, and to reduce tissue damage through inhibition of serine proteases released by polymorphonuclear neutrophil granulocytes (PMNs). Microarray analysis of bone marrow (BM) populations highly enriched in promyelocytes, myelocytes/metamyelocytes (MYs), and BM neutrophils demonstrates a transient, high mRNA expression of SLPI and genuine secondary granule proteins (GPs) in MYs. Consistent with this finding, immunostaining of BM cells showed SLPI and the secondary GP lactoferrin (LF) to be present in cells from the myelocyte stage and throughout neutrophil differentiation. Subcellular fractionation studies demonstrated the colocalization of SLPI and LF in subcellular fractions highly enriched in secondary granules. Finally, exocytosis studies demonstrated a corelease of SLPI and LF within minutes of activation. Collectively, these findings strongly indicate that SLPI is localized in secondary granules of PMNs. However, the amount of SLPI detected in PMNs is low compared with primary keratinocytes stimulated by growth factors involved in wound healing. This implicates that neutrophil-derived SLPI might not contribute essentially to re-establishment of homeostasis at sites of infection but rather, exert physiologically relevant intracellular activities. These might include the protection of secondary GPs against proteolytic activation and/or degradation by proteases, which might be dislocated to secondary granules at minute amounts as a consequence of spillover. PMID- 18285404 TI - Lack of CCR5 on dendritic cells promotes a proinflammatory environment in submandibular glands of the NOD mouse. AB - Sjogren's syndrome is an autoimmune disease characterized by lymphocytic infiltration of the salivary glands. In the NOD mouse, a model for this disease, the development of lymphocytic infiltrates in the salivary glands is preceded by an accumulation of dendritic cells (DC). Given the key importance of DC in regulating the immune response, we characterized the DC isolated from NOD salivary glands. These DC lacked membrane expression of CCR5, whereas DC from control salivary glands did express this molecule. The lack of expression was present already prior to the onset of lymphocytic infiltration, indicating that this was not the result of ongoing inflammation. DC from other sources in the NOD mouse also showed a decrease in CCR5 expression. The lack of CCR5 expression in the NOD salivary gland was accompanied by an increase in inflammatory chemokines. Furthermore, DC from CCR5-/- animals or DC treated with a CCR5 antagonist showed increased secretion of IL-12. Interestingly, in Sjogren's syndrome patients, CCR5 expression on circulating monocytes was decreased and correlated to increased levels of IL-12. These data indicate that CCR5 has regulatory properties and that the lack of CCR5 in NOD DC contributes to the proinflammatory environment in the salivary glands. PMID- 18285403 TI - MyD88 is necessary for neutrophil recruitment in hypersensitivity pneumonitis. AB - Hypersensitivity pneumonitis is an interstitial lung disease that is characterized by alveolitis, granuloma formation, and in some patients, fibrosis. Using the Saccharopolyspora rectivirgula animal model of Farmer's lung disease, our laboratory has demonstrated that neutrophils play a critical role in IFN gamma production during the acute phase of the disease. As IFN-gamma is necessary for granuloma formation, it is important to identify the factors that lead to neutrophil recruitment during disease. To begin to identify the pattern recognition receptors (PRRs) that initiate chemokine production, leading to neutrophil recruitment following S. rectivirgula exposure, we examined the role of MyD88 and TLR2. Our results demonstrate that neutrophil recruitment, as measured by flow cytometry and the myeloperoxidase assay, was abolished in the absence of MyD88 following S. rectivirgula exposure. The decrease in neutrophil recruitment was likely a result of a significant decrease in production of neutrophil chemokines MIP-2 and keratinocyte-derived chemokine. These results suggest that S. rectivirgula interacts with PRRs that are upstream of the MyD88 pathway to initiate cytokine and chemokine production. In vitro studies suggest that S. rectivirgula can interact with TLR2, and stimulation of adherent cells from TLR2 knockout (KO) mice with S. rectivirgula resulted in a significant decrease in MIP-2 production. However, TLR2 KO mice did not have a reduction in neutrophil recruitment compared with wild-type mice following S. rectivirgula exposure. The results from our studies suggest that one or more PRR(s) upstream of MyD88 are necessary for neutrophil recruitment following S. rectivirgula exposure. PMID- 18285405 TI - Lovastatin inhibits formation of AA amyloid. AB - Amyloid A (AA) amyloidosis is a severe complication of many chronic inflammatory disorders, including the hereditary periodic fever syndromes. However, in one of these periodic fever syndromes, the hyper IgD and periodic fever syndrome, amyloidosis is rare despite vigorous, recurring inflammation. This hereditary syndrome is caused by mutations in the gene coding for mevalonate kinase, an enzyme of the isoprenoid pathway. In this study, we used a cell culture system with human monocytes to show that inhibition of the isoprenoid pathway inhibits amyloidogenesis. Inhibition of the isoprenoid pathway by lovastatin resulted in a dose-dependent reduction of amyloid formed [53% at 10 microM (P=0.01)] compared with mononuclear cells that are exposed only to serum AA. The inhibitory effects of lovastatin are reversible by addition of farnesol but not geranylgeraniol. Farnesyl transferase inhibition also inhibited amyloidogenesis. These results implicate that the isoprenoid metabolism could be a potential target for prevention and treatment of AA amyloidosis. PMID- 18285406 TI - Tumor-infiltrating myeloid-derived suppressor cells are pleiotropic-inflamed monocytes/macrophages that bear M1- and M2-type characteristics. AB - Here, tumor-infiltrating CD11b(+) myelomonocytoid cells in murine colon adenocarcinoma-38 and GL261 murine glioma were phenotypically characterized. Over 90% were of the CD11b(+)F4/80(+) monocyte/macrophage lineage. They also had a myeloid-derived suppressor cell (MDSC) phenotype, as they suppressed the proliferation of activated splenic CD8(+) T cells and had a CD11b(+)CD11c(+)Gr 1(low)IL-4Ralpha(+) phenotype. In addition, the cells expressed CX(3)CR1 and CCR2 simultaneously, which are the markers of an inflammatory monocyte. The MDSCs expressed CD206, CXCL10, IL-1beta, and TNF-alpha mRNAs. They also simultaneously expressed CXCL10 and CD206 proteins, which are typical, classical (M1) and alternative (M2) macrophage activation markers, respectively. Peritoneal exudate cells (PECs) strongly expressed CD36, CD206, and TGF-beta mRNA, which is characteristic of deactivated monocytes. The MDSCs also secreted TGF-beta, and in vitro culture of MDSCs and PECs with anti-TGF-beta antibody recovered their ability to secrete NO. However, as a result of secretion of proinflammatory cytokines, MDSCs could not be categorized into deactivated monocyte/macrophages. Thus, tumor-infiltrating MDSCs bear pleiotropic characteristics of M1 and M2 monocytes/macrophages. Furthermore, CD206 expression by tumor-infiltrating MDSCs appears to be regulated by an autocrine mechanism that involves TGF-beta. PMID- 18285407 TI - The impact of risk factors and more stringent diagnostic criteria of gestational diabetes on outcomes in central European women. AB - OBJECTIVES: In the face of the ongoing discussion on the criteria for the diagnosis of gestational diabetes (GDM), we aimed to examine whether the criteria of the Fourth International Workshop Conference of GDM (WC) select women and children at risk better than the World Health Organization (WHO) criteria. DESIGN AND SETTING: This was a prospective longitudinal open study in five tertiary care centers in Austria. PATIENTS AND OUTCOME MEASURES: The impact of risk factors, different thresholds (WC vs. WHO), and numbers of abnormal glucose values (WC) during the 2-h, 75-g oral glucose tolerance test on fetal/neonatal complications and maternal postpartum glucose tolerance was studied in 1466 pregnant women. Women were treated if at least one value according to the WC (GDM-WC1) was met or exceeded. RESULTS: Forty-six percent of all women had GDM-WC1, whereas 29% had GDM-WHO, and 21% of all women had two or three abnormal values according to WC criteria (GDM-WC2). Eighty-five percent of the GDM-WHO were also identified by GDM-WC1. Previous GDM [odds ratio (OR) 2.9], glucosuria (OR 2.4), preconceptual overweight/obesity (OR 2.3), age 30 yr or older (OR 1.9), and large-for gestational age (LGA) fetus (OR 1.8) were the best independent predictors of the occurrence of GDM. Previous GDM (OR 4.4) and overweight/obesity (OR 4.0) also independently predicted diabetes postpartum. GDM-WC1 had a higher rate of obstetrical complications (LGA neonates, neonatal hypoglycemia, cesarean sections; P < 0.001) and impaired postpartum glucose tolerance (P < 0.0001) than GDM-WHO. CONCLUSION: These results suggest the use of more stringent WC criteria for the diagnosis of GDM with the initiation of therapy in case of one fasting or stimulated abnormal glucose value because these criteria detected more LGA neonates with hypoglycemia and mothers with impaired postpartum glucose metabolism than the WHO criteria. PMID- 18285408 TI - Increased androgen response to follicle-stimulating hormone administration in women with polycystic ovary syndrome. AB - CONTEXT: In women with polycystic ovary syndrome (PCOS), excess ovarian androgen production is driven by increased LH secretion. Studies conducted in animals suggest that the granulosa cell may influence LH-stimulated theca cell androgen production. OBJECTIVE: The objective of this study was to determine whether FSH enhances androgen production in women with PCOS compared with that of normal women. DESIGN: A prospective study was conducted to compare androgen production in response to FSH in two groups of women. SETTING: The study was conducted in a General Clinical Research Center in a tertiary academic medical center. PATIENTS: Women with PCOS, 18-35 yr (n = 20), and normal ovulatory controls, 18-35 yr (n = 10), were recruited for study. INTERVENTIONS: Serial blood samples were obtained over a 24-h period after an iv injection of recombinant human FSH (150 IU). MAIN OUTCOME MEASURES: The main outcome measures were serum 17-hydroxyprogesterone (17 OHP), androstenedione (A), dehydroepiandrosterone (DHEA), testosterone (T), and inhibin B (Inh B) responses after FSH administration. RESULTS: Basal serum 17 OHP, A, and T levels were markedly increased in women with PCOS compared with that observed in normal women. Basal DHEA and Inh B levels were similar to those of normal controls. After FSH injection, PCOS women demonstrated enhanced production of 17-OHP, A, DHEA, and Inh B, whereas in normal women no increases were observed. T levels declined slightly in both groups. CONCLUSIONS: These findings provide evidence that, in PCOS women, theca cell androgen production is enhanced by FSH administration and suggest a granulosa-theca cell paracrine mechanism. PMID- 18285409 TI - Cardiometabolic differences in children born after in vitro fertilization: follow up study. AB - CONTEXT: Increasing evidence suggests that adverse conditions during early prenatal life are associated with cardiometabolic dysfunction in postnatal life. In vitro fertilization (IVF) conception may be an early prenatal life event with long-term health consequences. OBJECTIVE: Our objective was to investigate several cardiometabolic measures in 8- to 18-yr-old IVF singletons and spontaneously conceived controls born from subfertile parents. DESIGN AND SETTING: This follow-up study was conducted at the VU University Medical Center, Amsterdam, The Netherlands. PARTICIPANTS: Blood pressure was examined in 225 IVF conceived children and 225 age- and gender-matched spontaneously conceived control children. Several indicators of insulin resistance were studied in a pubertal subpopulation (131 IVF children and 131 controls). MAIN OUTCOME MEASURES: Blood pressure, fasting glucose, and fasting insulin were determined. RESULTS: Systolic and diastolic blood pressure levels were higher in IVF children than controls (109 +/- 11 vs. 105 +/- 10 mm Hg, P < 0.001; and 61 +/- 7 vs. 59 +/ 7 mm Hg, P < 0.001, respectively). Children born after IVF were also more likely to be in the highest systolic and diastolic blood pressure quartiles (odds ratio = 2.1, 95% confidence interval 1.4, 3.3; odds ratio = 1.9, 95% confidence interval 1.2, 3.0, respectively). Furthermore, higher fasting glucose levels were observed in pubertal IVF children (5.0 +/- 0.4 vs. 4.8 +/- 0.4 mmol/liter in controls; P = 0.005). Blood pressure and fasting glucose differences could not be explained by current body size, birth weight, and other early life factors or by parental characteristics, including subfertility cause. CONCLUSIONS: These findings highlight the importance of continued cardiometabolic monitoring of IVF conceived children and might contribute to current knowledge about periconceptional influences and their consequences in later life. PMID- 18285410 TI - SOX2 plays a critical role in the pituitary, forebrain, and eye during human embryonic development. AB - CONTEXT: Heterozygous, de novo mutations in the transcription factor SOX2 are associated with bilateral anophthalmia or severe microphthalmia and hypopituitarism. Variable additional abnormalities include defects of the corpus callosum and hippocampus. OBJECTIVE: We have ascertained a further three patients with severe eye defects and pituitary abnormalities who were screened for mutations in SOX2. To provide further evidence of a direct role for SOX2 in hypothalamo-pituitary development, we have studied the expression of the gene in human embryonic tissues. RESULTS: All three patients harbored heterozygous SOX2 mutations: a deletion encompassing the entire gene, an intragenic deletion (c.70_89del), and a novel nonsense mutation (p.Q61X) within the DNA binding domain that results in impaired transactivation. We also show that human SOX2 can inhibit beta-catenin-driven reporter gene expression in vitro, whereas mutant SOX2 proteins are unable to repress efficiently this activity. Furthermore, we show that SOX2 is expressed throughout the human brain, including the developing hypothalamus, as well as Rathke's pouch, the developing anterior pituitary, and the eye. CONCLUSIONS: Patients with SOX2 mutations often manifest the unusual phenotype of hypogonadotropic hypogonadism, with sparing of other pituitary hormones despite anterior pituitary hypoplasia. SOX2 expression patterns in human embryonic development support a direct involvement of the protein during development of tissues affected in these individuals. Given the critical role of Wnt-signaling in the development of most of these tissues, our data suggest that a failure to repress the Wnt-beta-catenin pathway could be one of the underlying pathogenic mechanisms associated with loss-of-function mutations in SOX2. PMID- 18285411 TI - Association of pioglitazone treatment with decreased bone mineral density in obese premenopausal patients with polycystic ovary syndrome: a randomized, placebo-controlled trial. AB - OBJECTIVE: Our objective was to investigate the effect of pioglitazone on bone mineral density (BMD) and bone turnover markers in polycystic ovary syndrome (PCOS). DESIGN AND SETTING: We conducted a randomized, placebo-controlled study at an outpatient clinic at a university hospital. PATIENTS: Thirty premenopausal patients with PCOS and 14 age- and weight-matched healthy females participated. INTERVENTIONS: Pioglitazone (30 mg/d) or placebo was given for 16 wk. MAIN OUTCOME MEASURES: Measurements of BMD [hip (neck and total) and lumbar spine (L2 L4)], bone metabolic parameters [alkaline phosphatase (ALP), 25-hydroxyvitamin D, C-telopeptide of type I collagen (ICTP), osteocalcin, and PTH], endocrine profiles (testosterone, estradiol, and insulin), and body composition (waist to hip ratio, body mass index, and whole-body dual-energy x-ray absorptiometry scans) were performed. RESULTS: Patients with PCOS had significantly higher levels of ICTP, fasting insulin, and testosterone than controls, whereas no differences were measured in ALP, PTH, body composition, or BMD. Pioglitazone treatment was followed by reduced BMD [geometric means (-2 to +2 sd)]: lumbar spine 1.140 (0.964-1.348) vs. 1.127 (0.948-1.341) g/cm(2) (average decline 1.1%) and femoral neck 0.966 (0.767-1.217) vs. 0.952 (0.760-1.192) g/cm(2) (average decline 1.4%), both P < 0.05. Both ALP and PTH decreased significantly during pioglitazone treatment, whereas no significant changes were measured in 25 hydroxyvitamin D, ICTP, osteocalcin, sex hormones, and body composition. CONCLUSION: Pioglitazone treatment was followed by decreased lumbar and hip BMD and decreased measures of bone turnover in a premenopausal study population relatively protected from bone mineral loss. PMID- 18285412 TI - Single-nucleotide polymorphism rs7754840 of CDKAL1 is associated with impaired insulin secretion in nondiabetic offspring of type 2 diabetic subjects and in a large sample of men with normal glucose tolerance. AB - CONTEXT: CDKAL1 is a recently discovered susceptibility gene for type 2 diabetes. OBJECTIVE: Our objective was to investigate the impact of rs7754840 of CDKAL1 on insulin secretion, insulin sensitivity, and risk of type 2 diabetes. DESIGN AND SETTINGS: Study 1 (the EUGENE2 study) was a cross-sectional study including subjects from five white populations in Europe (Denmark, Finland, Germany, Italy, and Sweden). Study 2 is an ongoing prospective study of Finnish men. PARTICIPANTS: In study 1, 846 nondiabetic offspring of type 2 diabetic patients (age 40 +/- 10 yr; body mass index 26.7 +/- 5.0 kg/m(2)) participated. In study 2, subjects included 3900 middle-aged men (533 type 2 diabetic and 3367 nondiabetic subjects). INTERVENTIONS: INTERVENTIONS included iv glucose-tolerance test (IVGTT), oral glucose-tolerance test (OGTT), and euglycemic-hyperinsulinemic clamp in study 1 and OGTT in study 2. MAIN OUTCOME MEASURES: Parameters of insulin secretion, insulin resistance, and glucose tolerance status were assessed. RESULTS: In study 1, carriers of the GC and CC genotypes of rs7754840 had 11 and 24% lower first-phase insulin release in an IVGTT compared with that in carriers of the GG genotype (P = 0.002). The C allele was also associated with higher glucose area under the curve in an OGTT (P = 0.016). In study 2, rs7754840 was significantly associated with type 2 diabetes (P = 0.022) and markers of impaired insulin release [insulinogenic index (IGI), P = 0.012] in 2405 men with normal glucose tolerance. CONCLUSIONS: rs7754840 of CDKAL1 was associated with markers of impaired insulin secretion in two independent studies. Furthermore, rs7754840 was associated with type 2 diabetes in Finnish men (study 2). Therefore, CDKAL1 is likely to increase the risk of type 2 diabetes by impairing insulin secretion. PMID- 18285413 TI - Factors related to declining luteal function in women during the menopausal transition. AB - CONTEXT: Reproductive hormones are incompletely characterized during the menopause transition (MT). HYPOTHESIS: Increased anovulation and decreased progesterone accompany progress through the MT. DESIGN: The Daily Hormone Study (DHS) of the Study of Women's Health Across the Nation (SWAN) included 848 women aged 43-53 yr at baseline who collected daily urine for one cycle or up to 50 d annually for 3 yr. MAIN OUTCOME MEASURES: LH, FSH, estrone conjugates, and pregnanediol glucuronide levels were assessed. Cycles were classified by presumed luteal (ovulatory) status and bleeding. Hormones were related to time in study, age, menopausal status, and selected variables. RESULTS: Ovulatory-appearing cycles declined from 80.9% at baseline to 64.7% by the third assessment (H3). Cycles presumed anovulatory and not ending with bleeding by 50 d (anovulatory/nonbleeding) increased from 8.4 to 24% by H3 and were associated with progress to early perimenopause [odds ratio (OR) = 2.66; confidence interval (CI) = 1.17-6.04] or late perimenopause (OR = 56.21; CI = 18.79-168.12; P < 0.0001), African-American ethnicity (OR = 1.91; CI = 1.06-3.43), and less than high school education (OR = 3.51; CI = 1.62-7.62). Anovulatory cycles ending with bleeding remained at about 10% from baseline to H3; compared with ovulatory cycles, they were associated with obesity (OR = 4.68; CI = 1.33-16.52) and more than high school education (OR = 2.12; CI = 1.22-3.69; P = 0.02). Serum estradiol in both the highest and lowest categories was associated with anovulatory/nonbleeding collections. Pregnanediol glucuronide decreased 6.6% for each year on study. Insulin sensitivity measures did not relate strongly to menstrual cycle hormones. CONCLUSIONS: Anovulation without bleeding represents progression of the MT. A small but detectable decrease in luteal progesterone excretion occurs as women progress through the MT. PMID- 18285414 TI - Health status, mood, and cognition in experimentally induced subclinical thyrotoxicosis. AB - OBJECTIVE: Our objective was to determine whether subclinical thyrotoxicosis alters health status, mood, and/or cognitive function. DESIGN: This was a double blinded, randomized, cross-over study of usual dose l-T(4) (euthyroid arm) vs. higher dose l-T(4) (subclinical thyrotoxicosis arm) in hypothyroid subjects. PATIENTS: A total of 33 hypothyroid subjects receiving l-T(4) were included in the study. MEASUREMENTS: Subjects underwent measurements of health status, mood, and cognition: Short Form 36 (SF-36); Profile of Mood States (POMS); and tests of declarative memory (Paragraph Recall, Complex Figure), working memory (N-Back, Subject Ordered Pointing, and Digit Span Backwards), and motor learning (Pursuit Rotor). These were repeated after 12 wk on each of the study arms. RESULTS: Mean TSH levels decreased from 2.15 to 0.17 mU/liter on the subclinical thyrotoxicosis arm (P < 0.0001), with normal mean free T(4) and free T(3) levels. The SF-36 physical component summary and general health subscale were slightly worse during the subclinical thyrotoxicosis arm, whereas the mental health subscale was marginally improved. The POMS confusion, depression, and tension subscales were improved during the subclinical thyrotoxicosis arm. Motor learning was better during the subclinical thyrotoxicosis arm, whereas declarative and working memory measures did not change. This improvement was related to changes in the SF-36 physical component summary and POMS tension subscales and free T(3) levels. CONCLUSIONS: We found slightly impaired physical health status but improvements in measures of mental health and mood in l-T(4) treated hypothyroid subjects when subclinical thyrotoxicosis was induced in a blinded, randomized fashion. Motor learning was also improved. These findings suggest that thyroid hormone directly affects brain areas responsible for affect and motor function. PMID- 18285415 TI - Rickets in the Middle East: role of environment and genetic predisposition. AB - CONTEXT: The Middle East has a high incidence of rickets, and it is also common in Europe-dwelling children of Middle Eastern origin. OBJECTIVE: The objective of the study was to explore the mechanisms leading to rickets in children of the Middle East. DESIGN AND SETTING: We conducted a prospective study in 98 rachitic and 50 controls (aged 6 months to 4 yr) from university and community outpatient hospitals in Egypt and Turkey. MAIN OUTCOME MEASURES: We collected epidemiological, maternal, nutritional, radiographic, and biochemical parameters; markers of bone turnover; and vitamin D receptor (VDR) gene polymorphisms. RESULTS: Epidemiological factors had a key role in pursuit of rickets; Egyptian and Turkish patients had lower (P < 0.01) dietary calcium intake than controls and the recommended dietary intakes, and serum 25-hydroxyvitamin D levels were reduced in patients, the difference with controls being significant (P < 0.001) only in Turkey, although rickets was more severe in Egypt as determined by the x ray score (P < 0.05). In Turkey, the F VDR allele frequency was significantly (P < 0.05) increased in patients. The BB VDR genotype was associated with lower serum 25-hydroxyvitamin D levels in both patients and controls and with severity of rickets. CONCLUSIONS: In Turkey most patients had vitamin D deficiency, whereas in Egypt they had mostly calcium insufficiency combined with vitamin D deficiency. In this environ, VDR genotypes may predispose to rickets by increased frequency of the F allele. The unique environs and genetic predisposition have to be accounted for in the design of preventive measures, rather than using European or American recommended dietary intake for calcium and vitamin D. PMID- 18285416 TI - Paternal skeletal size predicts intrauterine bone mineral accrual. AB - BACKGROUND: We have previously demonstrated that maternal body build and lifestyle factors predict neonatal bone mineral accrual. However, the paternal determinants of neonatal bone mass are not known. In this study we explored the relationship between a father's bone mass and that of his offspring. METHODS: A total of 278 pregnancies (142 male and 136 female neonates) were recruited from the Southampton Women's Survey, a unique, well-established cohort of women, aged 20-34 yr, who had been assessed before and during pregnancy. The neonates and their fathers underwent whole body dual-x-ray absorptiometry (DXA) within 2 wk of birth using a Lunar DPX (General Electric Corp., Madison, WI) and Hologic Discovery instrument (Hologic Inc., Bedford, MA), respectively; correlation and regression methods were used to explore the parental determinants of neonatal bone mass. RESULTS: After adjusting the paternal DXA indices for father's age and the neonatal for baby's gestational age and age at DXA scan, there were highly significant positive associations between baby's whole body bone area, bone mineral content, and bone mineral density and the corresponding indices in the father (P = 0.003, 0.0002, 0.046, respectively) among female infants. These relationships were independent of maternal height and fat stores. The associations for male infants with paternal DXA indices did not achieve statistical significance. CONCLUSIONS: The father's skeletal size predicts skeletal size more strongly in female than male offspring, independently of the mother's body build. These data point toward the importance of considering paternal genotype in studies exploring the developmental origins of osteoporotic fracture and raise intriguing mechanistic questions about the gender specificity of influences on intrauterine bone mineral accrual. PMID- 18285417 TI - Circulating levels of inflammatory markers predict change in bone mineral density and resorption in older adults: a longitudinal study. AB - CONTEXT: IL-1, IL-6, and TNF-alpha play an important role in the pathogenesis of osteoporosis in animals; however, evidence that these play a similar role in bone loss in human studies is limited. OBJECTIVE: Our objective was to determine the associations between serum markers of inflammation and changes in bone mineral density (BMD) and urinary pyridinoline (PYR) to creatinine (Cr) ratio over 2.9 yr in older adults. METHODS: A total of 168 randomly selected subjects (mean 63 yr, range 52-78, 48% female) was studied. BMD was measured by dual-energy x-ray absorptiometry at baseline (mean T score: -0.18 to -0.61) and 2.9 yr later. Serum high-sensitivity (hs) C-reactive protein (CRP), IL-6, TNF-alpha, and the urinary PYR/Cr ratio were measured on both occasions. RESULTS: The mean annual loss of BMD was 0.15, 0.15, and 0.34% at total body, spine, and hip, respectively. Change in total body BMD was associated with baseline hs-CRP, IL-6, and TNF-alpha, as well as change in hs-CRP (beta: -0.41%/U, 95% confidence interval -0.68%, -0.15%) and IL-6 (beta: -0.62%/U, 95% confidence interval -1.01%, -0.23%). If these markers were put in the same predictive model, only IL-6 remained largely unchanged. Changes in other BMD sites were significantly predicted by IL-6 (hip and spine) and TNF-alpha (spine only). Finally, change in the PYR/Cr ratio was positively associated baseline IL-6, hs-CRP, and their changes (all P < 0.05) in women, but not men. CONCLUSIONS: Variation within the low levels of inflammatory markers observed in this study, especially IL-6, predicts bone loss and resorption, suggesting that targeted antiinflammatory therapy has potential for the prevention of osteoporosis. PMID- 18285418 TI - Dissociation between the insulin-sensitizing effect of rosiglitazone and its effect on hepatic and intestinal lipoprotein production. AB - CONTEXT: Despite its potent, well-documented insulin-sensitizing effects, rosiglitazone (RSG) does not effectively ameliorate the hypertriglyceridemia of insulin-resistant or diabetic individuals and has even been shown to slightly but significantly increase triglyceride-rich lipoproteins (TRL) in some studies. The mechanism of this effect is currently not known. OBJECTIVE: We investigated the effect of RSG treatment on TRL metabolism. DESIGN: This was a 12-wk, single sequence, cross-over study of rosiglitazone vs. placebo for 6 wk. PARTICIPANTS: Participants included 17 nondiabetic men with a broad range of insulin sensitivity. INTERVENTION: INTERVENTION included rosiglitazone 8 mg/d vs. placebo for 6 wk. MAIN OUTCOME MEASURE: TRL metabolism (concentration, production and catabolic rates) was assessed in a constant fed state with a 12-h primed constant infusion of [D3]l-leucine and multicompartmental modeling. RESULTS: RSG treatment resulted in significant insulin sensitization with no change in body weight. Fasting plasma triglyceride (TG) concentration, however, was higher with RSG vs. placebo (P = 0.0006), as were fasting and fed TRL-TG, TRL-apoB-48, and TRL-apoB 100 (fed TRL-apoB-48: 0.93 +/- 0.08 vs. 0.76 +/- 0.07 mg/dl, P =0.017, and fed TRL-apoB-100: 15.57 +/- 0.90 vs. 13.71 +/- 1.27 mg/dl, P = 0.029). This small but significant increase in plasma TRL concentration was explained by a tendency for RSG to increase TRL production and reduce particle clearance, as indicated by the significantly increased production to clearance ratios for both apoB-48 containing (0.43 +/- 0.03 vs. 0.34 +/- 0.03, P = 0.048) and apoB-100-containing (7.0 +/- 0.4 vs. 6.2 +/- 0.6, P = 0.029) TRL. CONCLUSION: These data indicate dissociation between the insulin-sensitizing effects of RSG and absence of anticipated reductions in production rates of apoB-100- and apoB-48-containing TRL particles, which may explain the absence of TG lowering seen in humans treated with this agent. PMID- 18285419 TI - The influence of gestational stage on urinary iodine excretion in pregnancy. AB - INTRODUCTION: Median urinary iodine concentration (UIC) is the most commonly used indicator of population iodine nutrition. However, its validity as an indicator of dietary intake relies on a stable relationship between dietary iodine intake and urinary excretion. Physiological alterations in normal pregnancy, such as increased glomerular filtration rate, potentially invalidate UIC as an assessment tool in pregnancy. OBJECTIVE: The objective of the study was to document the impact of advancing gestation on UIC in normal pregnancy and determine whether the current reference intervals for general population iodine monitoring are appropriate for use in the context of pregnancy. DESIGN: Tasmania has a well described history of mild iodine deficiency (school-age median UIC of 84 microg/liter). We assessed UIC in 759 urine samples from 431 women attending the Antenatal Clinic at the Royal Hobart Hospital, Tasmania's primary teaching hospital. MAIN OUTCOME: The overall median UIC during pregnancy was 75 microg/liter (95% confidence interval 70.03-79.97 microg/liter) at a median gestation of 19.4 wk. Stratification by gestation, however, revealed a dynamic relationship between ioduria and gestation. Median UIC was elevated in early pregnancy and subsequently declined with advancing gestation. CONCLUSION: In this mildly iodine-deficient population, current reference intervals for UIC overestimated the adequacy of iodine nutrition during the first and early second trimester of pregnancy. Gestation-specific UIC reference intervals are required to classify iodine nutrition during pregnancy. This is particularly important in populations with borderline iodine deficiency. PMID- 18285420 TI - Spermatogenic and steroidogenic impairment of the testicle characterizes the hereditary leucine-75-proline apolipoprotein a-I amyloidosis. AB - CONTEXT: The leucine-75-proline variant of apolipoprotein A-I leads to a new hereditary systemic amyloidosis involving mostly the liver and kidney. OBJECTIVE: The objective of the study was to examine the effects of this amyloidosis on testicular structure and function. DESIGN: This was an observational study in which patients with testicular amyloidosis were characterized. SETTING: The study was carried out at the Endocrinology Department of Brescia University. PATIENTS OR OTHER PARTICIPANTS: Over a 13-yr period, 25 patients were found to be affected by leucine-75-proline apolipoprotein A-I testicular amyloidosis. Thirteen had the testicle as the first or only organ involved (group 1); in 12 testicular damage followed that of other organs (group 2). INTERVENTIONS: There were no interventions. MAIN OUTCOME MEASURE: Hormone and lipidic profiles, semen analysis, echographic volume of testicles, testicular histology, and genetic analysis were carried out. RESULTS: Group 1 patients were younger than those of group 2. In group 1, eight had hypergonadotropic hypogonadism and five were normogonadic with high gonadotropins; in group 2 all subjects were hypogonadic. All men had low high-density lipoprotein values. Group 1 patients were macroorchid, whereas the testicular volume was at the highest limit in group 2 (group 1 vs. group 2, P < 0.05). All men in the first group and six in the second group were azoospermic; the remaining had oligoposia. Biopsies showed the germinal epithelium replaced by amyloid. Leydig cells were essentially preserved in normogonadic but not hypogonadic patients. CONCLUSIONS: This amyloidosis may determine infertility, macroorchidism, and hypogonadism. Endocrine impairment follows spermatogenic impairment. PMID- 18285421 TI - Activation of the estrogen receptor contributes to the progression of pulmonary lymphangioleiomyomatosis via matrix metalloproteinase-induced cell invasiveness. AB - CONTEXT: The role of estrogens in the pathogenesis of lymphangioleiomyomatosis (LAM), an aggressive and destructive, eventually fatal lung disease of women, is poorly understood. OBJECTIVE: The study was conducted to test the hypothesis that the lung disease in LAM is estrogen mediated and to determine whether estrogens contribute to the invasiveness of LAM. DESIGN: In vitro cell culture of spindle shaped LAM cells (LAMD-SM) were isolated and propagated from affected lungs. Estrogen receptor (ER)-alpha and ERbeta analyses were conducted by RT-PCR. ERalpha and ERbeta, tissue inhibitor of metalloproteinase-2, and matrix metalloproteinases (MMP)-2 had Western blot analysis for protein assessment. Activity assays were performed for MT1-MMP, MMP-2, and tissue inhibitor of metalloproteinase-2. Assessment of MMP-2 promoter function was done via transfection assays. Cell invasion chambers were used to determine and quantitate cell invasiveness. SETTING: The study was conducted at an academic medical center. PATIENTS: Tissue and cells were obtained from patients as outlined in approved institution review board protocol (97/007). INTERVENTION: LAMD-SM cells were treated with a specific MMP-2 antibody or a nonspecific inhibitor, doxycycline. MAIN OUTCOME MEASURES: Activity of MMP-2 and invasiveness of LAMD-SM cells were measured. RESULTS: LAMD-SM cells express functional ERs (ERalpha and ERbeta), which undergo rapid intracellular turnover in their unbound state. 17beta-estradiol (E(2)) enhances the transcriptional ER activity. E(2)-induced ER activation increases synthesis and activity of MMP-2 through posttranscriptional mechanisms in LAMD-SM. The E(2)/ER-mediated increase of MMP-2 promotes LAMD-SM invasiveness, in assays in vitro, which can be inhibited by specific antibodies against MMP-2 or doxycycline, an inhibitor of MMPs. CONCLUSION: The invasion and destruction of lung parenchyma in LAM is, at least partially, an estrogen-MMP driven process, which has major implications for therapeutic interventions. PMID- 18285422 TI - Pancreatic glucose uptake in vivo in men with newly diagnosed type 1 diabetes. AB - CONTEXT: Due to the restricted accessibility of pancreatic tissue in living man, direct analysis of the events preceding development of autoimmune changes in the pancreas has been problematic. In vivo imaging of insulitis might markedly increase understanding of the events and timing of the events that are necessary for the progression toward overt type 1 diabetes. DESIGN: To evaluate possibilities to visualize insulitis in man in vivo with positron emission tomography, we studied 12 male patients (age 26 +/- 7 yr) with newly diagnosed type 1 diabetes (duration range 0-7 months) and nine age- and sex-matched healthy controls after an overnight fast using 2-[(18)F]fluoro-2-deoxy-D-glucose and [(11)C]methionine. For definition of the regions of interest, pancreas was localized with magnetic resonance imaging or computed tomography-positron emission tomography. RESULTS: Glucose uptake to the pancreas was markedly higher in the patients with type 1 diabetes than in the healthy controls (22.9 +/- 6.4 vs. 17.8 +/- 6.0 micromol/kg.min, P = 0.039). Glucose uptake to the pancreas of the patients was inversely associated with the duration of diabetes (r = -0.58; P = 0.024), so that in patients with newly diagnosed type 1 diabetes, glucose uptake was higher than in the healthy controls or patients with long duration of diabetes. Methionine uptake to the pancreas of the patients was similar as in the controls (3.7 +/- 1.9 vs. 4.6 +/- 2.4 micromol/kg.min, P = 0.21). CONCLUSIONS: In patients with type 1 diabetes, glucose uptake to the pancreas is enhanced at or soon after the time of diagnosis. PMID- 18285423 TI - Identification of a novel PEX14 mutation in Zellweger syndrome. AB - BACKGROUND: Peroxisome biogenesis disorders are a clinically and genetically heterogeneous group of very severe autosomal recessive disorders caused by impaired peroxisome biogenesis. The prototype of this group of disorders is the cerebro-hepato-renal syndrome of Zellweger. METHODS AND RESULTS: Here we report a patient with Zellweger syndrome, who presented at the age of 3 months with icterus, dystrophy, axial hypotonia, facial dysmorphy, posterior embryotoxon, and hepatomegaly. Abnormal findings of metabolic screening tests included hyperbilirubinaemia, hypoketotic dicarboxylic aciduria, increased C(26:0) and decreased C(22:0) plasma levels, and strongly reduced plasmalogen concentrations. In fibroblasts, both peroxisomal alpha- and beta-oxidation were impaired. Liver histology revealed bile duct paucity, cholestasis, arterial hyperplasia, very small branches of the vena portae, and parenchymatic destruction. Immunocytochemical analysis of cultured fibroblasts demonstrated that the cells contain peroxisomal remnants lacking apparent matrix protein content and PEX14, a central membrane component of the peroxisomal matrix protein import machinery. Transfection of fibroblasts with a plasmid coding for wild-type PEX14 restored peroxisomal matrix protein import, indicating that the primary genetic defect affecting the patient is indeed linked to PEX14. Mutational analysis of this gene revealed a genomic deletion leading to the deletion of exon 3 from the coding DNA (c.85-?_170+?del) and a concomitant change of the reading frame (p.[Ile29_Lys56del;Gly57GlyfsX2]). CONCLUSIONS: This report represents the second PEX14-deficiency associated with Zellweger syndrome and the first documentation of a PEX14-deficient patient with detailed clinical follow-up and biochemical, morphological, and radiological data. PMID- 18285424 TI - Interferon regulatory factor 5 (IRF5) gene variants are associated with multiple sclerosis in three distinct populations. AB - BACKGROUND: IRF5 is a transcription factor involved both in the type I interferon and the toll-like receptor signalling pathways. Previously, IRF5 has been found to be associated with systemic lupus erythematosus, rheumatoid arthritis and inflammatory bowel diseases. Here we investigated whether polymorphisms in the IRF5 gene would be associated with yet another disease with features of autoimmunity, multiple sclerosis (MS). METHODS: We genotyped nine single nucleotide polymorphisms and one insertion-deletion polymorphism in the IRF5 gene in a collection of 2337 patients with MS and 2813 controls from three populations: two case-control cohorts from Spain and Sweden, and a set of MS trio families from Finland. RESULTS: Two single nucleotide polymorphism (SNPs) (rs4728142, rs3807306), and a 5 bp insertion-deletion polymorphism located in the promoter and first intron of the IRF5 gene, showed association signals with values of p<0.001 when the data from all cohorts were combined. The predisposing alleles were present on the same common haplotype in all populations. Using electrophoretic mobility shift assays we observed allele specific differences in protein binding for the SNP rs4728142 and the 5 bp indel, and by a proximity ligation assay we demonstrated increased binding of the transcription factor SP1 to the risk allele of the 5 bp indel. CONCLUSION: These findings add IRF5 to the short list of genes shown to be associated with MS in more than one population. Our study adds to the evidence that there might be genes or pathways that are common in multiple autoimmune diseases, and that the type I interferon system is likely to be involved in the development of these diseases. PMID- 18285425 TI - High frequency of genomic deletions--and a duplication--in the LIS1 gene in lissencephaly: implications for molecular diagnosis. AB - BACKGROUND: LIS1 is the main gene causing classical (isolated) lissencephaly predominating in the posterior brain regions (p>a). However, about 40% of patients with this malformation pattern show no abnormality after fluorescence in situ hybridisation (FISH) analysis of the 17p13.3 region and LIS1 sequencing. To investigate whether alternative gene(s) or genomic deletions/duplications of LIS1 may account for the high percentage of individuals who show no abnormalities on FISH and sequencing, we performed multiplex ligation dependent probe amplification assay (MLPA) in a series of patients. METHODS: We initially performed DNA sequencing in 45 patients with isolated lissencephaly with a p>a gradient, in whom FISH had revealed normal results. We subsequently performed MLPA in those who were mutation negative, and long range polymerase chain reaction (PCR) to characterise the breakpoint regions in patients in whom the deletions were small enough. RESULTS: We found LIS1 mutations in 44% of patients (20/45) of the whole sample and small genomic deletions/duplications in 76% of the remaining (19/25). Deletions were much more frequent than duplications (18 vs 1). Overall, small genomic deletions/duplications represented 49% (19/39) of all LIS1 alterations and brought to 87% (39/45) the number of patients in whom any involvement of LIS1 could be demonstrated. Breakpoint characterisation, performed in 5 patients, suggests that Alu mediated recombination is a major molecular mechanism underlying LIS1 deletions. CONCLUSIONS: LIS1 is highly specific for isolated p>a lissencephaly. The high frequency of genomic deletions/duplications of LIS1 is in keeping with the over representation of Alu elements in the 17p13.3 region. MLPA has a high diagnostic yield and should be used as first line molecular diagnosis for p>a lissencephaly. PMID- 18285426 TI - Molecular characterisation of SMARCB1 and NF2 in familial and sporadic schwannomatosis. AB - BACKGROUND: Schwannomatosis is a rare condition characterised by multiple schwannomas and lack of involvement of the vestibular nerve. A recent report identified bi-allelic mutations in the SMARCB1/INI1 gene in a single family with schwannomatosis. We aimed to establish the contribution of the SMARCB1 and the NF2 genes to sporadic and familial schwannomatosis in our cohort. METHODS: We performed DNA sequence and dosage analysis of SMARCB1 and NF2 in 28 sporadic cases and 15 families with schwannomatosis. RESULTS: We identified germline mutations in SMARCB1 in 5 of 15 (33.3%) families with schwannomatosis and 2 of 28 (7.1%) individuals with sporadic schwannomatosis. In all individuals with a germline mutation in SMARCB1 in whom tumour tissue was available, we detected a second hit with loss of SMARCB1. In addition, in all affected individuals with SMARCB1 mutations and available tumour tissue, we detected bi-allelic somatic inactivation of the NF2 gene. SMARCB1 mutations were associated with a higher number of spinal tumours in patients with a positive family history (p = 0.004). CONCLUSION: In contrast to the recent report where no NF2 mutations were identified in a schwannomatosis family with SMARCB1 mutations, in our cohort, a four hit model with mutations in both SMARCB1 and NF2 define a subset of patients with schwannomatosis. PMID- 18285427 TI - A missense mutation in PTCH2 underlies dominantly inherited NBCCS in a Chinese family. AB - BACKGROUND: Naevoid basal cell carcinoma syndrome (NBCCS) is a pleiotropic, autosomal dominant disease. Growing evidence suggests that the disorder may result from mutations in genes of the Sonic hedgehog (Shh) signalling pathway. OBJECTIVE: To investigate the pathogenic gene in a Chinese Han family with NBCCS. METHODS: Mapping and mutation screening were used to investigate the candidate genes SHH, PTCH, PTCH2 and SMO. A GLI1 reporter gene and a cell growth curve were used to examine functional consequences of the detected mutant. RESULTS: One novel mutation, a G-->A transition (2157G-->A) in exon 15 of the PTCH2 gene, was identified in this family with NBCCS by direct sequencing and digestion with the AvaI restriction enzyme. The mutation was not found in normal family members or in 520 controls. The mutation led to an R719Q amino acid substitution in an extracellular loop of the PTCH2 protein. Functional studies revealed that the R719Q mutation resulted in inactivation of PTCH2 inhibitory activities. In contrast to wild type PTCH2, PTCH2-R719Q could not inhibit cell proliferation. CONCLUSION: PTCH2 (2157G-->A), a novel missense mutation, underlies NBCCS, resulting in the loss of PTCH2 inhibitory function in the Shh signalling pathway. PMID- 18285428 TI - The protective effect of farm animal exposure on childhood allergy is modified by NPSR1 polymorphisms. AB - BACKGROUND: Little is known about the asthma candidate gene neuropeptide S receptor 1 (NPSR1) in relation to environmental exposures, but recent evidences suggest its role as an effect modifier. OBJECTIVES: To explore the interaction between NPSR1 polymorphisms and environmental exposures related to farming lifestyle and to study the in vitro effects of lipopolysaccharide (LPS) stimulation on NPSR1 expression levels. METHODS: We studied 3113 children from PARSIFAL, a European cross-sectional study on environmental/lifestyle factors and childhood allergy, partly focused on children brought up on a farm. Information on exposures and outcomes was primarily obtained from parental questionnaires. Seven tagging polymorphisms were analysed in a conserved haplotype block of NPSR1. Multivariate logistic regression was used to evaluate a multiplicative model of interaction. NPSR1 protein and messenger RNA (mRNA) levels in monocytes were measured after LPS stimulation by fluorescence activated cell sorting (FACS) and quantitative real-time polymerase chain reaction (PCR). RESULTS: A strong interaction was seen between current regular contact to farm animals and several NPSR1 polymorphisms, particularly rs323922 and rs324377 (p<0.005), with respect to allergic symptoms. Considering the timing of initiation of such current regular farm animal contact, significant interactions with these and two additional polymorphisms (SNP546333, rs740347) were revealed. In response to LPS, NPSR1-A protein levels in monocytes were upregulated (p = 0.002), as were NPSR1-A mRNA levels (p = 0.02). CONCLUSIONS: The effect of farm animal contact on the development of allergic symptoms in children is modified by NPSR1 genetic background. PMID- 18285429 TI - Comparison of the effect of two types of acupuncture on quality of life in secondary progressive multiple sclerosis: a preliminary single-blind randomized controlled trial. AB - OBJECTIVE: To compare the effect of two types of acupuncture on the quality of life of individuals with secondary progressive multiple sclerosis and provide preliminary evidence regarding the safety of this intervention for this population. DESIGN: Preliminary single-blind randomized controlled trial. SETTING: Outpatient attendance at rehabilitation unit. PARTICIPANTS: Fourteen participants with secondary progressive multiple sclerosis. INTERVENTIONS: Chinese medical acupuncture or minimal acupuncture. Participants received 10 treatments over five weeks. MEASURES: Multiple Sclerosis Impact Scale 29, Fatigue Severity Scale and General Health Questionnaire 12 were measured pre and post intervention. Adverse events and other responses during treatment were recorded prospectively. RESULTS: Participants receiving minimal acupuncture demonstrated statistically significant greater improvement in the Multiple Sclerosis Impact Scale 29 psychological subscale compared with those receiving Chinese medical acupuncture in an intention-to-treat analysis (P=0.04), with mean change in Chinese acupuncture group of 6.0 (SD 13.9) and in minimal acupuncture group of 23.0 (SD 21.0). No other statistically significant difference between the groups was found. No major adverse events were noted. Minor adverse events such as lower limb muscle spasms or pain were noted in some participants in both intervention groups. CONCLUSION: Minimal acupuncture resulted in greater improvement of Multiple Sclerosis Impact Scale 29 psychological subscale compared with Chinese medical acupuncture. No other differences between the groups were found. In view of the small sample these results are not conclusive. This study provides preliminary evidence to suggest that acupuncture is safe for people with secondary progressive multiple sclerosis. A large-scale trial is required to provide more definitive evidence. PMID- 18285430 TI - Long-term benefits of exercising on quality of life and fatigue in multiple sclerosis patients with mild disability: a pilot study. AB - OBJECTIVE: To determine if exercise benefits patients with multiple sclerosis. DESIGN: Randomized controlled trial. SETTINGS: Participants exercised at home and also attended exercise classes held in a hospital physiotherapy gym. SUBJECTS: Thirty patients, diagnosed and independently mobile, were recruited in the Dublin area. INTERVENTION: For three months, classes were held twice-weekly and participants exercised independently once-weekly. The control group was monitored monthly and management remained unchanged. MEASUREMENTS: Measurements were taken at baseline, three and six months. The Modified Fatigue Impact Scale (MFIS), Multiple Sclerosis Impact Scale-29 (MSIS-29) and Functional Assessment of Multiple Sclerosis (FAMS) were used to measure fatigue and quality of life (QOL). Heart rate (HR) and the Borg's Rating of Perceived Exertion (RPE) were recorded during an incremental exercise test. The change from baseline scores between groups was compared using the Mann-Whitney U-test. RESULTS: Twenty-four participants completed the programme (n = 12 in each group). Based on the change in scores at three months, the exercise group had significantly greater improvements in exercise capacity (HR: -14 [-18.5, -2.5] versus 0.5 [-4, 5.5], P= 0.009), QOL (FAMS: 23 [9.5, 42.5] versus -3.5 [-16, 5], P=0.006) and fatigue (MFIS: -13 [-20, -3] versus 1 [-4, 4.5], P=0.02). At six months, the difference in change scores remained significant for FAMS (19 [14, 31] versus -4.5 [-25, 8], P=0.002) and MFIS (-8.5 [-19.5, -1] versus 0.5 [-2.5, 6.5], P=0.02) only. CONCLUSIONS: A three-month exercise programme improved participants' exercise capacity, QOL and fatigue, with the improvements in QOL and fatigue lasting beyond the programme. PMID- 18285431 TI - How feasible is the attainment of community ambulation after stroke? A pilot randomized controlled trial to evaluate community-based physiotherapy in subacute stroke. AB - OBJECTIVE: This pilot randomized controlled trial evaluated an assistant-led, community-based intervention to improve community mobility and participation after stroke, and examined the potential for independent community ambulation in people with subacute stroke who present with moderate gait deficit. DESIGN: A multicentre, pilot randomized controlled trial. SETTING: Three hospitals and three community settings in New Zealand. SUBJECTS: Thirty post-acute, home dwelling stroke survivors were randomly allocated to receive intervention in the community (n = 14) or as hospital outpatients (n = 16) twice a week for seven weeks. INTERVENTIONS: The community intervention involved practice of functional gait activities in community environments relevant to each participant. Hospital based physiotherapy was based upon a Motor Relearning approach. MAIN MEASURES: The primary outcome measure was gait speed (m/min). Secondary outcomes included endurance (six-minute walk time), Activities-specific Confidence Balance Scale, and the Subjective Index of Physical and Social Outcomes measured at baseline, post intervention and six months. RESULTS: Large gains in gait speed were obtained for participants in both groups: community group mean (SD) 16 (16.1) m/min; physiotherapy group mean (SD) 15.9 (16.1) m/min, maintained at six months. There were no significant differences between groups for primary and secondary outcomes after treatment (P = 0.86 ANOVA) or at six months (P = 0.83 ANOVA). Only 11 participants reported independent community ambulation. Levels of social integration were low to moderate. CONCLUSIONS: A community-based gait recovery programme appears a practicable alternative to routine physiotherapy, however independent community ambulation is a challenging rehabilitation goal. PMID- 18285432 TI - Multiple-task walking training in people with mild to moderate Parkinson's disease: a pilot study. AB - OBJECTIVE: To determine whether multiple-task training of walking is feasible and worthwhile in people with mild to moderate Parkinson's disease. DESIGN: Baseline controlled pilot study with three-week baseline, training and retention phases. SETTING: University research laboratory. SUBJECTS: Five people aged 61 +/-8 years with mild to moderate Parkinson's disease (Hoehn and Yahr stages II-III). INTERVENTIONS: Multiple-task training was performed for 30 minutes once a week during the three-week training phase. MAIN MEASURES: Participants were tested and trained during the 'on' phase of medication. Feasibility of training was examined using 10 cm visual analogue scales to measure participant perception of mental fatigue, physical fatigue, difficulty, anxiety and confidence. Walking velocity, stride length and cadence under multiple-task conditions were measured. RESULTS: Participants reported low levels of mental fatigue, physical fatigue, difficulty and anxiety as well as high levels of confidence associated with multiple-task training. No adverse effects of training were reported. Multiple-task walking velocity increased by 0.09 +/- 0.06 m/s (95% confidence interval (CI) 0.02 to 0.16) between the baseline and training phase and this increase was maintained in the retention phase. CONCLUSION: Multiple-task training is feasible in people with mild to moderate Parkinson's disease. The data suggest sustained benefits on multiple-task walking velocity are possible. A randomized controlled trial of multiple-task training efficacy is warranted. PMID- 18285433 TI - Assessing clinically meaningful change following a programme for managing chronic pain. AB - OBJECTIVES: (1) To identify criteria for clinically meaningful change scores in commonly used measures of pain and related disability, using Goal Attainment Scaling as an external indicator of success. (2) To investigate the chances of achieving these scores following a pain management programme versus remaining on the waiting list. DESIGN: Participants were assessed on entry to waiting list, on admission and at follow-up from the pain management programme. Three groups of Goal Attainment Scaling scores (-1.0, +1) were created from final Goal Attainment Scaling achievements. Mean scores on other measures were analysed in relation to Goal Attainment Scaling score groups by ANOVA. Differences in numbers achieving clinically meaningful changes when on the pain management programme or waiting list were compared. SUBJECTS: Chronic musculoskeletal pain participants (N = 73) attending a rehabilitation centre, mean age (range) 44.8 (24-70) years, mean age (range) 44.8 (24-70), mean pain duration 7.7 (1-32) years. INTERVENTION: Three week (15-day) pain management programme based on cognitive behavioural principles. MEASURES: McGill Pain Questionnaire, 0-10 Pain Intensity Numerical Rating Scale (NRS), Oswestry Disability Questionnaire (ODQ), General Health Questionnaire (GHQ), 5 minute walk, 1 minute sit/stand, 1 minute stair-climbing, and Goal Attainment Scaling (GAS). RESULTS: Mean change scores differed significantly for three GAS groups and were highest in the most successful (+1) group. These scores were used to define clinically meaningful changes on the NRS (-3), ODQ (-12), walk (+87) and stairs (+14). Significantly more participants on the pain management programme achieved these scores than those on the waiting list. CONCLUSION: Using GAS as a criterion of patient-perceived improvement enabled identification of clinically meaningful changes on some other common measures. These successfully differentiated achievement between patients on the pain management programme and those on the waiting list. PMID- 18285434 TI - An exploratory study of two measures of free-living physical activity for people with multiple sclerosis. AB - OBJECTIVE: To examine the ability of two measures of physical activity (tri-axial accelerometer and activity diary) to discriminate among groups of inactive, moderately active and active individuals with multiple sclerosis and to explore the relationship between these two measures. DESIGN: Exploratory, descriptive study. SUBJECTS: Thirty individuals with multiple sclerosis and nine controls. PROTOCOL: Individuals with multiple sclerosis were recruited to inactive, moderately active and active groups as defined by Adjusted Activity Scores from the Human Activity Profile. Control participants were recruited to an active group. Free-living physical activity was recorded over four consecutive days. MAIN MEASURES: A TriTrac RT3 accelerometer and a self-report physical activity diary were used to measure activity. RESULTS: Thirty-six participants completed data collection. For the participants with multiple sclerosis, both the accelerometer (P = 0.004) and the diary (P = 0.006) detected significant differences between inactive and active groups. The accelerometer also detected a significant difference between moderately active and active groups (P = 0.04). In contrast, the diary detected a significant difference between inactive and moderately active groups (P = 0.05). Accelerometer and diary scores were significantly correlated (r = 0.59). Accelerometer scores were significantly correlated with neurological status (r(s) = -0.64). CONCLUSIONS: Both measures readily differentiated least active from most active groups. The accelerometer also differentiated moderately active from active groups, suggesting suitability for use in detecting change in more active client groups, while the diary differentiated inactive from moderately active groups, suggesting suitability for use in detecting change in less active groups. PMID- 18285435 TI - Physical limitations and perceived quality of care among family medicine patients. AB - OBJECTIVE: To assess how physical limitations relate to patient satisfaction with the quality of medical care among family medicine patients, after adjustment for self-rated overall health and other patient characteristics. DESIGN: A telephone survey of family medicine patients, linked with information extracted from medical records. SETTING: A large family medicine department in Rochester, Minnesota. SUBJECTS: Patient ratings of quality were linked to medical records for 674 adult patients. RESULTS: Adjusting for self-rated overall health status and other confounders using multiple logistic regression analysis revealed that having physical limitations was inversely and independently related to excellent perceived quality (adjusted odds ratio = 0.56, P= 0.035). CONCLUSIONS: In our sample of family medicine patients, people with physical limitations experience a disparity in perceived quality of care. PMID- 18285436 TI - Two simple approaches for validating a binary surrogate endpoint using data from multiple trials. AB - A surrogate endpoint is an endpoint that is observed before a true endpoint and is used to draw conclusions about the effect of intervention on true endpoint. To gauge confidence in the use of a surrogate endpoint, it must be validated. Two simple validation methods using data from multiple trials with surrogate and true endpoints are discussed: an estimation method extending previous work and new method based on hypothesis tests. The validation methods were applied to two data sets, each involving 10 randomized trials: one for patients with early colon cancer where the true endpoint was survival status at eight years and surrogate endpoint was cancer recurrence status at three years, and one for patients with advanced colorectal cancer where the true endpoint was survival status at 12 months and the surrogate endpoint was cancer recurrence status at six months. The estimation method uses the surrogate endpoint in the new trial and a model relating surrogate and true endpoints in previous trials to predict the effect of intervention on true endpoint in the new trial. For validation, each trial was successively treated as the ;new' trial and a comparison was made between predicted and observed effects of intervention on true endpoint. Performance of the surrogate endpoint was good in both data sets. The hypothesis testing method involves the z-statistic for the surrogate endpoint, which is the estimated effect of intervention on surrogate endpoint divided by its standard error. To use this z-statistic to test a null hypothesis of no effect of intervention on true endpoint, the critical value is increased above a standard level of 1.96 to a level determined by the relationships between surrogate and true endpoints in the data sets. This elevated critical value could be used for accelerated approval. PMID- 18285437 TI - Use of progression-free survival as a surrogate marker in oncology trials: some regulatory issues. AB - There has been interest in using progression-free survival as a surrogate endpoint for overall survival in oncology clinical trials. In order to objectively define this endpoint, clear understanding of what progression means, how it is measured and what its implications are need to be discussed. This article discusses some regulatory aspects of using progression-free survival as an endpoint. PMID- 18285438 TI - Surrogate endpoint validation: statistical elegance versus clinical relevance. AB - A variety of approaches have been proposed to provide formal and informal validation of proposed surrogate markers. To achieve true clinical impact, the validation must convince both the statistical and clinical communities. In this paper, we argue that the best approach is not a single method but a multi-faceted exploration, using multiple approaches, including those that directly appeal to clinicians but with less statistical foundation and those arising from statistical considerations but more difficult to interpret clinically. We illustrate our approach using data from clinical trials in both early and advanced colorectal cancer. PMID- 18285439 TI - Individual- and trial-level surrogacy in colorectal cancer. AB - Two conditions must be fulfilled for an intermediate endpoint to be an acceptable surrogate for a true clinical endpoint: (1) there must be a strong association between the surrogate and the true endpoint, and (2) there must be a strong association between the effects of treatment on the surrogate and the true endpoint. We test whether these conditions are fulfilled for disease-free survival (DFS) and progression-free survival (PFS) on data from 20 clinical trials comparing experimental treatments with standard treatments for early and advanced colorectal cancer. The effects of treatment on DFS (or PFS in advanced disease) and OS were quantified through log hazard ratios (log HR), estimated through a Weibull model stratified for trial. The rank correlation coefficients between DFS and OS, and trial-specific treatment effects, were estimated using a bivariate copula distribution for these endpoints. A linear regression model between the estimated log hazard ratios was used to compute the "surrogate threshold effect", which is the minimum treatment effect on DFS required to predict a non-zero treatment effect on OS in a future trial. In early disease, the rank correlation coefficient between DFS and OS was equal to 0.96 (CI 0.95 0.97). The correlation coefficient between the log hazard ratios was equal to 0.94 (CI 0.87-1.01). The risk reductions were approximately 3% smaller on OS than on DFS, and the surrogate threshold effect corresponded to a DFS hazard ratio of 0.93. In advanced disease, the rank correlation coefficient between PFS and OS was equal to 0.82 (CI 0.82-0.83). The correlation coefficient between the log hazard ratios was equal to 0.99 (CI 0.94-1.04). The risk reductions were approximately 19% smaller on OS than on PFS, and the surrogate threshold effect corresponded to a PFS hazard ratio of 0.86. One trial with a large treatment effect on PFS and OS had a strong influence on the results in advanced disease. DFS (and PFS in advanced disease) are acceptable surrogates for OS in colorectal cancer. PMID- 18285440 TI - Practical issues arising in an exploratory analysis evaluating progression-free survival as a surrogate endpoint for overall survival in advanced colorectal cancer. AB - This paper is based on a conference presentation in which several authors presented results from analyses of the same dataset concerning the evaluation of progression-free survival (PFS) as a surrogate endpoint for overall survival in advanced colorectal cancer clinical trials. In evaluating a potential surrogate endpoint, there is a hierarchy of information that might usually be considered desirable: 1) a biological rationale for surrogacy, 2) demonstration of the prognostic value of the surrogate endpoint in untreated patients and 3) in treated patients and 4) demonstration across randomized comparisons that differences in the effect of randomized treatments on the surrogate endpoint are associated with the corresponding differences in the effect on the clinical endpoint of interest. Results from analyses that might be used to address the third and four requirements are presented and some of the practical issues that arise in evaluating a surrogate endpoint, which would be relevant to many diseases, are illustrated. Although the results presented should not be seen as a definitive analysis of the value of PFS as a surrogate endpoint, concerns are identified about the potential lack of standardization of the definition of PFS or the frequency of evaluation of disease progression and the high leverage of one study in evaluating the association in addressing the fourth requirement. PMID- 18285441 TI - Surrogate endpoints for overall survival in advanced colorectal cancer: a clinician's perspective. AB - Surrogate endpoints in oncology research and practice have garnered increasing attention over the past two decades. This activity has largely been driven by the promise surrogate endpoints appear to hold: the potential to get new therapies to seriously ill patients more rapidly. However, uncertainties abound. Even agreeing upon a definition of a "valid" surrogate endpoint has not been a straightforward exercise; this article begins by highlighting differences in how this term has been previously captured and applied, as well as laying out the basic criteria essential for its application in advanced colorectal cancer. Ideally, these elements include (but are not limited to) ease of measurement, rapid indication of treatment effect, and, most importantly, reliable and consistent prediction of the true impact of a treatment on the ultimate outcome of interest: overall survival. The strengths and weaknesses of current potential surrogate endpoints in advanced colorectal cancer, including performance status, carcinoembryonic antigen plasma level, overall response rate, time to progression, and disease free survival, are each considered in turn. Finally, limitations of surrogate endpoints in the clinical setting, including challenges in extrapolation to new therapies, and the incomplete provision of information about potential adverse effects, are discussed. Work remains to be done between physicians and statisticians to bridge the gap between that which is statistically demonstrable and that which will be clinically useful.The term ;surrogate endpoint' was virtually unknown by most oncologists 15 years ago. A search in PubMed [http://www.ncbi.nlm.nih.gov] based on the words ;surrogate and cancer' shows that more than 2000 papers were published in medical journals in the last 20 years, with a dramatic increase of interest in the last five years. Interestingly, the same trend is observed when the words ;surrogate and heart' are entered into PubMed, suggesting that the issue of surrogate endpoints goes beyond the field of oncology, although the frequency of discussion varies (Figure 1; note different y-axis scales for oncology and cardiology).The goal of the present paper is to discuss the main issues surrounding surrogate endpoints from a clinician's point of view, using as an example surrogate endpoints of overall survival (OS) in advanced colorectal cancer (ACC). PMID- 18285442 TI - Surrogate endpoints for overall survival in early colorectal cancer from the clinician's perspective. AB - The administration of adjuvant chemotherapy after resection of stage III colon cancer to prolong disease-free survival (DFS) and increase overall survival (OS) has been clinical standard since the early 1990s. Recently, 3-year DFS was recognized as surrogate endpoint for OS based on a meta-analysis of trials utilizing 5-fluorouracil as only active chemotherapy component. The standard of care in adjuvant therapy, however, has moved on to modern combination regimens including oxaliplatin, and novel targeted agents such as angiogenesis inhibitors and antibodies against epidermal growth factor receptor are currently undergoing rigorous testing in phase III adjuvant trials. For the practicing clinician, the use of surrogate endpoints to appreciate the efficacy of a specific adjuvant therapy contains various challenges, in particular, in discussions with patients. It is questionable whether 3-year DFS can still be considered an appropriate predictor of OS in complex clinical scenarios with continuous change in treatment standards in the adjuvant and palliative situation. Thus, the practicing oncologist needs to be aware of the limitations in the definition of surrogate endpoints in the adjuvant setting. PMID- 18285443 TI - Evaluating time to cancer recurrence as a surrogate marker for survival from an information theory perspective. AB - The last two decades have seen a lot of development in the area of surrogate marker validation. One of these approaches places the evaluation in a meta analytic framework, leading to definitions in terms of trial- and individual level association. A drawback of this methodology is that different settings have led to different measures at the individual level. Using information theory, Alonso et al. proposed a unified framework, leading to a new definition of surrogacy, which offers interpretational advantages and is applicable in a wide range of situations. In this work, we illustrate how this information-theoretic approach can be used to evaluate surrogacy when both endpoints are of a time-to event type. Two meta-analyses, in early and advanced colon cancer, respectively, are then used to evaluate the performance of time to cancer recurrence as a surrogate for overall survival. PMID- 18285444 TI - The PKCdelta -Abl complex communicates ER stress to the mitochondria - an essential step in subsequent apoptosis. AB - Conditions that compromise protein folding in the endoplasmic reticulum trigger the unfolded protein response (UPR), which either restores proper protein folding or results in cellular demise through apoptosis. In this study, we found that, in response to ER stress in vivo and in vitro, PKCdelta translocates to the ER where it binds to the tyrosine kinase Abl. Tyrosine phosphorylation and kinase activity of PKCdelta are required for PKCdelta binding to Abl in the ER. Moreover, we found that inhibition of PKCdelta by the PKCdelta-specific peptide inhibitor deltaV1-1 or by silencing of PKCdelta reduces ER-stress-induced JNK activation and inhibits ER-stress-mediated apoptosis. Furthermore, the inhibitor of PKCdelta kinase activity rottlerin blocks the translocation of the PKCdelta-Abl complex from the ER to the mitochondria and confers protection against apoptosis. Thus, PKCdelta communicates ER stress to the mitochondria by binding to ER-localized Abl. The PKCdelta-Abl complex then translocates to the mitochondria, communicating ER stress to this organelle, thereby, triggering apoptosis. PMID- 18285445 TI - Ca2+-store-dependent and -independent reversal of Stim1 localization and function. AB - Stim1 responds to depletion of ER Ca2+ stores by rearranging from tubular structures throughout the ER into punctate structures near the plasma membrane, where it activates Orai store-operated Ca2+ entry (SOCE) channels. However, the mechanism and structural determinants of the localization and reversal of Stim1 puncta formation are poorly understood. Using HEK293 cells expressing Stim1 tagged with enhanced yellow fluorescent protein (EYFP-Stim1), we show that the basis for SOCE termination is the reversal of the punctate Stim1 localization, which absolutely depends on SOCE-dependent store refilling. We also describe rapid, store-independent reversal of EYFP-Stim1 punctae by the ML-9 inhibitor of myosin-light-chain kinase (MLCK). ML-9 similarly inhibited SOCE and the Ca2+ release-activated Ca2+ (CRAC) current. Reversal by ML-9 resulted in full re establishment of the tubular EYFP-Stim1 localization. A constitutively active EF hand mutant of EYFP-Stim1 was also reversed by ML-9, regardless of the Ca2+ store content. Inhibition by ML-9 was not due to MLCK inhibition as other inhibitors of MLCK had no effect. Finally, we provide evidence that EYFP-Stim1 punctae form in specific predetermined cellular loci. We conclude that SOCE is tightly coupled to formation of Stim1 puncta, and both SOCE and puncta formation involve a dynamic, reversible signaling complex that probably consists of components in addition to Stim1 and Orai channels. PMID- 18285446 TI - LRP1 controls biosynthetic and endocytic trafficking of neuronal prion protein. AB - The trafficking of normal cellular prion protein (PrPC) is believed to control its conversion to the altered conformation (designated PrPSc) associated with prion disease. Although anchored to the membrane by means of glycosylphosphatidylinositol (GPI), PrPC on neurons is rapidly and constitutively endocytosed by means of coated pits, a property dependent upon basic amino acids at its N-terminus. Here, we show that low-density lipoprotein receptor-related protein 1 (LRP1), which binds to multiple ligands through basic motifs, associates with PrPC during its endocytosis and is functionally required for this process. Moreover, sustained inhibition of LRP1 levels by siRNA leads to the accumulation of PrPC in biosynthetic compartments, with a concomitant lowering of surface PrPC, suggesting that LRP1 expedites the trafficking of PrPC to the neuronal surface. PrPC and LRP1 can be co-immunoprecipitated from the endoplasmic reticulum in normal neurons. The N-terminal domain of PrPC binds to purified human LRP1 with nanomolar affinity, even in the presence of 1 muM of the LRP specific chaperone, receptor-associated protein (RAP). Taken together, these data argue that LRP1 controls both the surface, and biosynthetic, trafficking of PrPC in neurons. PMID- 18285447 TI - Extracellular matrix retention of thrombospondin 1 is controlled by its conserved C-terminal region. AB - Thrombospondins (TSPs) are an evolutionarily ancient family of extracellular calcium-binding glycoproteins. The five mammalian TSPs collectively have important roles in angiogenesis and vascular biology, synaptogenesis, wound repair and connective tissue organisation. Their complex functions relate to the multiple postsecretion fates of TSPs that can involve endocytic uptake, proteolysis or retention within the extracellular matrix (ECM). Surprisingly, the molecular and cellular mechanisms by which TSPs become retained within the ECM are poorly understood. We hypothesised that the highly conserved TSP C-terminal domain mediates ECM retention. We report that ECM incorporation as insoluble punctate deposits is an evolutionarily conserved property of TSPs. ECM retention of TSP1 is mediated by the C-terminal region in trimeric form, and not by C terminal monomer or trimers of the N-terminal domain or type 1 repeats. Using a novel mRFP-tagged TSP1 C-terminal trimer, we demonstrate that ECM retention involves the RGD site and a novel site in the L-lectin domain with structural similarity to the ligand-binding site of cargo transport proteins. CD47 and beta1 integrins are dispensable for ECM retention, but beta1 integrins enhance activity. These novel data advance concepts of the molecular processes that lead to ECM retention of TSP1. PMID- 18285448 TI - Regulation of PI3K signalling by the phosphatidylinositol transfer protein PITPalpha during axonal extension in hippocampal neurons. AB - Phosphatidylinositol transfer proteins (PITPs) mediate the transfer of phosphatidylinositol (PtdIns) or phosphatidylcholine (PtdCho) between two membrane compartments, thereby regulating the interface between signalling, phosphoinositide (PI) metabolism and membrane traffic. Here, we show that PITPalpha is enriched in specific areas of the postnatal and adult brain, including the hippocampus and cerebellum. Overexpression of PITPalpha, but not PITPbeta or a PITPalpha mutant deficient in binding PtdIns, enhances laminin dependent extension of axonal processes in hippocampal neurons, whereas knockdown of PITPalpha protein by siRNA suppresses laminin and BDNF-induced axonal growth. PITPalpha-mediated axonal outgrowth is sensitive to phosphoinositide 3-kinase (PI3K) inhibition and shows dependency on the Akt/GSK-3/CRMP-2 pathway. We conclude that PITPalpha controls the polarized extension of axonal processes through the provision of PtdIns for localized PI3K-dependent signalling. PMID- 18285449 TI - Signal-dependent export of GABA transporter 1 from the ER-Golgi intermediate compartment is specified by a C-terminal motif. AB - The C-terminus of GABA transporter 1 (GAT1, SLC6A1) is required for trafficking of the protein through the secretory pathway to reach its final destination, i.e. the rim of the synaptic specialization. We identified a motif of three hydrophobic residues (569VMI571) that was required for export of GAT1 from the ER Golgi intermediate compartment (ERGIC). This conclusion was based on the following observations: (i) GAT1-SSS, the mutant in which 569VMI571 was replaced by serine residues, was exported from the ER in a COPII-dependent manner but accumulated in punctate structures and failed to reach the Golgi; (ii) under appropriate conditions (imposing a block at 15 degrees C, disruption of COPI), these structures also contained ERGIC53; (iii) the punctae were part of a dynamic compartment, because it was accessible to a second anterograde cargo [the temperature-sensitive variant of vesicular stomatitis virus G protein (VSV-G)] and because GAT1-SSS could be retrieved from the punctate structures by addition of a KKxx-based retrieval motif, which supported retrograde transport to the ER. To the best of our knowledge, the VMI-motif of GAT1 provides the first example of a cargo-based motif that specifies export from the ERGIC. PMID- 18285450 TI - Galpha12 regulates protein interactions within the MDCK cell tight junction and inhibits tight-junction assembly. AB - The polarized functions of epithelia require an intact tight junction (TJ) to restrict paracellular movement and to separate membrane proteins into specific domains. TJs contain scaffolding, integral membrane and signaling proteins, but the mechanisms that regulate TJs and their assembly are not well defined. Galpha12 (GNA12) binds the TJ protein ZO-1 (TJP1), and Galpha12 activates Src to increase paracellular permeability via unknown mechanisms. Herein, we identify Src as a component of the TJ and find that recruitment of Hsp90 to activated Galpha12 is necessary for signaling. TJ integrity is disrupted by Galpha12 stimulated Src phosphorylation of ZO-1 and ZO-2 (TJP2); this phosphorylation leads to dissociation of occludin and claudin 1 from the ZO-1 protein complex. Inhibiting Hsp90 with geldanamycin blocks Galpha12-stimulated Src activation and phosphorylation, but does not affect protein levels or the Galpha12-ZO-1 interaction. Using the calcium-switch model of TJ assembly and GST-TPR (GST-fused TPR domain of PP5) pull-downs of activated Galpha12, we demonstrate that switching to normal calcium medium activates endogenous Galpha12 during TJ assembly. Thrombin increases permeability and delays TJ assembly by activating Galpha12, but not Galpha13, signaling pathways. These findings reveal an important role for Galpha12, Src and Hsp90 in regulating the TJ in established epithelia and during TJ assembly. PMID- 18285451 TI - Stress-induced recruitment of epiplakin to keratin networks increases their resistance to hyperphosphorylation-induced disruption. AB - Epiplakin is a large (>725 kDa) cytoskeletal protein exclusively expressed in epithelial tissues. It has a unique structure, consisting entirely of plakin repeat domains (PRDs), one of the hallmarks of spectraplakin protein family members. Previous studies, including the phenotypic analyses of knockout mice, failed to reveal the biological function of epiplakin. Using in vitro binding assays, we show here that all but one of the 16 PRDs of mouse epiplakin bind to keratins of basal keratinocytes. Nevertheless, in primary keratinocyte cell cultures, epiplakin only partially colocalized with keratin intermediate filament networks. However, upon application of cellular stress in the form of keratin hyperphosphorylation, osmotic shock or UV irradiation, the entire cytoplasmic epiplakin pool became associated with keratin. In response to such types of stress, epiplakin initially translocated to the still-intact keratin filament network and remained associated with keratin after its disruption and transformation into granular aggregates. Time-course experiments revealed that serine/threonine (okadaic acid) and tyrosine (orthovanadate) phosphatase inhibitor-induced filament disruption in differentiated keratinocytes proceeded faster in epiplakin-deficient cells compared with wild-type cells. Our data suggest that epiplakin plays a role in keratin filament reorganization in response to stress, probably by protecting keratin filaments against disruption in a chaperone-like fashion. PMID- 18285452 TI - Degradation of a polyadenylated rRNA maturation by-product involves one of the three RRP6-like proteins in Arabidopsis thaliana. AB - Yeast Rrp6p and its human counterpart, PM/Scl100, are exosome-associated proteins involved in the degradation of aberrant transcripts and processing of precursors to stable RNAs, such as the 5.8S rRNA, snRNAs, and snoRNAs. The activity of yeast Rrp6p is stimulated by the polyadenylation of its RNA substrates. We identified three RRP6-like proteins in Arabidopsis thaliana: AtRRP6L3 is restricted to the cytoplasm, whereas AtRRP6L1 and -2 have different intranuclear localizations. Both nuclear RRP6L proteins are functional, since AtRRP6L1 complements the temperature-sensitive phenotype of a yeast rrp6Delta strain and mutation of AtRRP6L2 leads to accumulation of an rRNA maturation by-product. This by-product corresponds to the excised 5' part of the 18S-5.8S-25S rRNA precursor and accumulates as a polyadenylated transcript, suggesting that RRP6L2 is involved in poly(A)-mediated RNA degradation in plant nuclei. Interestingly, the rRNA maturation by-product is a substrate of AtRRP6L2 but not of AtRRP6L1. This result and the distinctive subcellular distribution of AtRRP6L1 to -3 indicate a specialization of RRP6-like proteins in Arabidopsis. PMID- 18285453 TI - A glycine-arginine domain in control of the human MRE11 DNA repair protein. AB - Human MRE11 is a key enzyme in DNA double-strand break repair and genome stability. Human MRE11 bears a glycine-arginine-rich (GAR) motif that is conserved among multicellular eukaryotic species. We investigated how this motif influences MRE11 function. Human MRE11 alone or a complex of MRE11, RAD50, and NBS1 (MRN) was methylated in insect cells, suggesting that this modification is conserved during evolution. We demonstrate that PRMT1 interacts with MRE11 but not with the MRN complex, suggesting that MRE11 arginine methylation occurs prior to the binding of NBS1 and RAD50. Moreover, the first six methylated arginines are essential for the regulation of MRE11 DNA binding and nuclease activity. The inhibition of arginine methylation leads to a reduction in MRE11 and RAD51 focus formation on a unique double-strand break in vivo. Furthermore, the MRE11 methylated GAR domain is sufficient for its targeting to DNA damage foci and colocalization with gamma-H2AX. These studies highlight an important role for the GAR domain in regulating MRE11 function at the biochemical and cellular levels during DNA double-strand break repair. PMID- 18285454 TI - RalGDS couples growth factor signaling to Akt activation. AB - The Akt kinase is a key regulator of cell proliferation and survival. It is activated in part by PDK1-induced phosphorylation. Here we show that RalGDS, a Ras effector protein that activates Ral GTPases, has a second function that promotes Akt phosphorylation by PDK1 by bringing these two kinases together. In support of this conclusion is our finding that suppression of RalGDS expression in cells inhibits both epidermal growth factor and insulin-induced phosphorylation of Akt. Moreover, while PDK1 complexes with N-GDS, Akt complexes with the central region of RalGDS through an intermediary, JIP1. The biological significance of this newly discovered RalGDS function is highlighted by the observation that an N-terminally deleted mutant of RalGDS that retains the ability to activate Ral proteins but loses the ability to activate Akt also fails to promote cell proliferation. Thus, RalGDS forms a nexus that transduces growth factor signaling to both Ral GTPase and Akt-mediated signaling cascades. PMID- 18285455 TI - Activation of FoxM1 during G2 requires cyclin A/Cdk-dependent relief of autorepression by the FoxM1 N-terminal domain. AB - The Forkhead transcription factor FoxM1 is an important regulator of gene expression during the G(2) phase. Here, we show that FoxM1 transcriptional activity is kept low during G(1)/S through the action of its N-terminal autoinhibitory domain. We found that cyclin A/cdk complexes are required to phosphorylate and activate FoxM1 during G(2) phase. Deletion of the N-terminal autoinhibitory region of FoxM1 generates a mutant of FoxM1 (DeltaN-FoxM1) that is active throughout the cell cycle and no longer depends on cyclin A for its activation. Mutation of two cyclin A/cdk sites in the C-terminal transactivation domain leads to inactivation of full-length FoxM1 but does not affect the transcriptional activity of the DeltaN-FoxM1 mutant. We show that the intramolecular interaction of the N- and C-terminal domains depends on two RXL/LXL motifs in the C terminus of FoxM1. Mutation of these domains leads to a similar gain of function as deletion of the N-terminal repressor domain. Based on these observations we propose a model in which FoxM1 is kept inactive during the G(1)/S transition through the action of the N-terminal autorepressor domain, while phosphorylation by cyclin A/cdk complexes during G(2) results in relief of inhibition by the N terminus, allowing activation of FoxM1-mediated gene transcription. PMID- 18285457 TI - Spatial interplay between PIASy and FIP200 in the regulation of signal transduction and transcriptional activity. AB - The members of the protein inhibitor of activated STAT (PIAS) family of proteins are implicated in fundamental cellular processes, including transcriptional regulation, either through action as E3 SUMO ligases or through SUMO-independent effects. We report here the identification of FIP200 (focal adhesion kinase family-interacting protein of 200 kDa) as a new PIASy-interacting protein. We show that the interaction depends on the integrity of the RING finger of PIASy and the carboxy terminus of FIP200. Both in vitro and in vivo sumoylation assays failed to reveal any sumoylation of FIP200, suggesting that FIP200 is not a bona fide SUMO substrate. Immunofluorescence microscopy and subcellular fractionation, either upon forced PIASy expression or in the absence of PIASy, revealed that interaction with PIASy redistributes FIP200 from the cytoplasm to the nucleus, correlating with abrogation of FIP200 regulation of TSC/S6K signaling. Conversely, FIP200 enhances the transcriptional activation of the p21 promoter by PIASy whereas PIASy transcription activity is severely reduced upon FIP200 depletion by RNA interference. Chromatin immunoprecipitation analysis demonstrates that endogenous PIASy and FIP200 are corecruited to the p21 promoter. Altogether, these results provide the first evidence for the existence of a close-spatially controlled-mode of regulation of FIP200 and PIASy nucleocytoplasmic functions. PMID- 18285456 TI - Hypoxia-inducible factor-dependent degeneration, failure, and malignant transformation of the heart in the absence of the von Hippel-Lindau protein. AB - Hypoxia-inducible transcription factor 1 (HIF-1) and HIF-2alpha regulate the expression of an expansive array of genes associated with cellular responses to hypoxia. Although HIF-regulated genes mediate crucial beneficial short-term biological adaptations, we hypothesized that chronic activation of the HIF pathway in cardiac muscle, as occurs in advanced ischemic heart disease, is detrimental. We generated mice with cardiac myocyte-specific deletion of the von Hippel-Lindau protein (VHL), an essential component of an E3 ubiquitin ligase responsible for suppressing HIF levels during normoxia. These mice were born at expected frequency and thrived until after 3 months postbirth, when they developed severe progressive heart failure and premature death. VHL-null hearts developed lipid accumulation, myofibril rarefaction, altered nuclear morphology, myocyte loss, and fibrosis, features seen for various forms of human heart failure. Further, nearly 50% of VHL(-/-) hearts developed malignant cardiac tumors with features of rhabdomyosarcoma and the capacity to metastasize. As compelling evidence for the mechanistic contribution of HIF-1alpha, the concomitant deletion of VHL and HIF-1alpha in the heart prevented this phenotype and restored normal longevity. These findings strongly suggest that chronic activation of the HIF pathway in ischemic hearts is maladaptive and contributes to cardiac degeneration and progression to heart failure. PMID- 18285458 TI - Splicing factors SF1 and U2AF associate in extraspliceosomal complexes. AB - Splicing factors SF1 and U2AF associate cooperatively with pre-mRNA and play a crucial role in 3' splice site recognition during early steps of spliceosome assembly. Formation of the active spliceosome subsequently displaces SF1 in a remodeling process that stabilizes the association of U2 snRNP with pre-mRNA. Fluorescence microscopy shows SF1 and U2AF distributed throughout the nucleoplasm, where transcription occurs, with additional concentration in nuclear speckles, where splicing factors accumulate when not engaged in splicing. Fluorescence recovery after photobleaching analysis in live cells shows that the mobilities of SF1 and the two subunits of U2AF (U2AF(65) and U2AF(35)) are correlated with the abilities of these proteins to interact with each other. Direct binding of SF1 to U2AF(65) was demonstrated by fluorescence resonance energy transfer in both the nucleoplasm and nuclear speckles. This interaction persisted after transcription inhibition, suggesting that SF1 associates with U2AF in a splicing-independent manner. We propose that SF1 and U2AF form extraspliceosomal complexes before and after taking part in the assembly of catalytic spliceosomes. PMID- 18285459 TI - The von Hippel-Lindau tumor suppressor protein and Egl-9-Type proline hydroxylases regulate the large subunit of RNA polymerase II in response to oxidative stress. AB - Human renal clear cell carcinoma (RCC) is frequently associated with loss of the von Hippel-Lindau (VHL) tumor suppressor (pVHL), which inhibits ubiquitylation and degradation of the alpha subunits of hypoxia-inducible transcription factor. pVHL also ubiquitylates the large subunit of RNA polymerase II, Rpb1, phosphorylated on serine 5 (Ser5) within the C-terminal domain (CTD). A hydroxylated proline 1465 within an LXXLAP motif located N-terminal to the CTD allows the interaction of Rpb1 with pVHL. Here we report that in RCC cells, pVHL regulates expression of Rpb1 and is necessary for low-grade oxidative-stress induced recruitment of Rpb1 to the DNA-engaged fraction and for its P1465 hydroxylation, phosphorylation, and nondegradative ubiquitylation. Egln-9-type prolyl hydroxylases, PHD1 and PHD2, coimmunoprecipitated with Rpb1 in the chromatin fraction of VHL(+) RCC cells in response to oxidative stress, and PHD1 was necessary for P1465 hydroxylation while PHD2 had an inhibitory effect. P1465 hydroxylation was required for oxidative-stress-induced Ser5 phosphorylation of Rpb1. Importantly, overexpression of wild-type Rpb1 stimulated formation of kidney tumors by VHL(+) cells, and this effect was abolished by P1465A mutation of Rpb1. These data indicate that through this novel pathway involving P1465 hydroxylation and Ser5 phosphorylation of Rbp1, pVHL may regulate tumor growth. PMID- 18285460 TI - Human Rvb1/Tip49 is required for the histone acetyltransferase activity of Tip60/NuA4 and for the downregulation of phosphorylation on H2AX after DNA damage. AB - The role of chromatin-remodeling factors in transcription is well established, but the link between chromatin-remodeling complexes and DNA repair remains unexplored. Human Rvb1 and Rvb2 are highly conserved AAA(+) ATP binding proteins that are part of various chromatin-remodeling complexes, such as Ino80, SNF2 related CBP activator protein (SRCAP), and Tip60/NuA4 complexes, but their molecular function is unclear. The depletion of Rvb1 increases the amount and persistence of phosphorylation on chromatin-associated H2AX after the exposure of cells to UV irradiation or to mitomycin C, cisplatin, camptothecin, or etoposide, without increasing the amount of DNA damage. Tip60 depletion, but not Ino80 or SRCAP depletion, mimics the effect of Rvb1 depletion on H2AX phosphorylation. Rvb1 is required for the histone acetyltransferase (HAT) activity of the Tip60 complex, and histone H4 acetylation is required prior to the dephosphorylation of phospho-H2AX. Thus, Rvb1 is critical for the dephosphorylation of phospho-H2AX due to the role of Rvb1 in maintaining the HAT activity of Tip60/NuA4, implicating the Rvb1-Tip60 complex in the chromatin-remodeling response of cells after DNA damage. PMID- 18285461 TI - A role for interferon regulatory factor 4 in receptor editing. AB - Receptor editing is the primary means through which B cells revise antigen receptors and maintain central tolerance. Previous studies have demonstrated that interferon regulatory factor 4 (IRF-4) and IRF-8 promote immunoglobulin light chain rearrangement and transcription at the pre-B stage. Here, the roles of IRF 4 and -8 in receptor editing were analyzed. Our results show that secondary rearrangement was impaired in IRF-4 but not IRF-8 mutant mice, suggesting that receptor editing is defective in the absence of IRF-4. The role of IRF-4 in receptor editing was further examined in B-cell-receptor (BCR) transgenic mice. Our results show that secondary rearrangement triggered by membrane-bound antigen was defective in the IRF-4-deficient mice. Our results further reveal that the defect in secondary rearrangement is more severe at the immunoglobulin lambda locus than at the kappa locus, indicating that IRF-4 is more critical for the lambda rearrangement. We provide evidence demonstrating that the expression of IRF-4 in immature B cells is rapidly induced by self-antigen and that the reconstitution of IRF-4 expression in the IRF-4 mutant immature B cells promotes secondary rearrangement. Thus, our studies identify IRF-4 as a nuclear effector of a BCR signaling pathway that promotes secondary rearrangement at the immature B-cell stage. PMID- 18285463 TI - Phosphoinositide 3-kinases p110alpha and p110beta regulate cell cycle entry, exhibiting distinct activation kinetics in G1 phase. AB - Phosphoinositide 3-kinase (PI3K) is an early signaling molecule that regulates cell growth and cell cycle entry. PI3K is activated immediately after growth factor receptor stimulation (at the G(0)/G(1) transition) and again in late G(1). The two ubiquitous PI3K isoforms (p110alpha and p110beta) are essential during embryonic development and are thought to control cell division. Nonetheless, it is presently unknown at which point each is activated during the cell cycle and whether or not they both control S-phase entry. We found that p110alpha was activated first in G(0)/G(1), followed by a minor p110beta activity peak. In late G(1), p110alpha activation preceded that of p110beta, which showed the maximum activity at this time. p110beta activation required Ras activity, whereas p110alpha was first activated by tyrosine kinases and then further induced by active Ras. Interference with p110alpha and -beta activity diminished the activation of downstream effectors with different kinetics, with a selective action of p110alpha in blocking early G(1) events. We show that inhibition of either p110alpha or p110beta reduced cell cycle entry. These results reveal that PI3Kalpha and -beta present distinct activation requirements and kinetics in G(1) phase, with a selective action of PI3Kalpha at the G(0)/G(1) phase transition. Nevertheless, PI3Kalpha and -beta both regulate S-phase entry. PMID- 18285462 TI - Posttranslational modification of the AU-rich element binding protein HuR by protein kinase Cdelta elicits angiotensin II-induced stabilization and nuclear export of cyclooxygenase 2 mRNA. AB - The mRNA stabilizing factor HuR is involved in the posttranscriptional regulation of many genes, including that coding for cyclooxygenase 2 (COX-2). Employing RNA interference technology and actinomycin D experiments, we demonstrate that in human mesangial cells (hMC) the amplification of cytokine-induced COX-2 by angiotensin II (AngII) occurs via a HuR-mediated increase of mRNA stability. Using COX-2 promoter constructs with different portions of the 3' untranslated region of COX-2, we found that the increase in COX-2 mRNA stability is attributable to a distal class III type of AU-rich element (ARE). Likewise, the RNA immunoprecipitation assay showed AngII-induced binding of HuR to this ARE. Using the RNA pulldown assay, we demonstrate that the AngII-caused HuR assembly with COX-2 mRNA is found in free and cytoskeleton-bound polysomes indicative of an active RNP complex. Mechanistically, the increased HuR binding to COX-2-ARE by AngII is accompanied by increased nucleocytoplasmic HuR shuttling and depends on protein kinase Cdelta (PKCdelta), which physically interacts with nuclear HuR, thereby promoting its phosphorylation. Mapping of phosphorylation sites identified serines 221 and 318 as critical target sites for PKCdelta-triggered HuR phosphorylation and AngII-induced HuR export to the cytoplasm. Posttranslational modification of HuR by PKCdelta represents an important novel mode of HuR activation implied in renal COX-2 regulation. PMID- 18285464 TI - Ezh2 requires PHF1 to efficiently catalyze H3 lysine 27 trimethylation in vivo. AB - The mammalian Polycomblike protein PHF1 was previously shown to interact with the Polycomb group (PcG) protein Ezh2, a histone methyltransferase whose activity is pivotal in sustaining gene repression during development and in adulthood. As Ezh2 is active only when part of the Polycomb Repressive Complexes (PRC2-PRC4), we examined the functional role of its interaction with PHF1. Chromatin immunoprecipitation experiments revealed that PHF1 resides along with Ezh2 at Ezh2-regulated genes such as the HoxA loci and the non-Hox MYT1 and WNT1 genes. Knockdown of PHF1 or of Ezh2 led to up-regulated HoxA gene expression. Interestingly, depletion of PHF1 did correlate with reduced occupancy of Bmi-1, a PRC1 component. As expected, knockdown of Ezh2 led to reduced levels of its catalytic products H3K27me2/H3K27me3. However, reduced levels of PHF1 also led to decreased global levels of H3K27me3. Notably, the levels of H3K27me3 decreased while those of H3K27me2 increased at the up-regulated HoxA loci tested. Consistent with this, the addition of PHF1 specifically stimulated the ability of Ezh2 to catalyze H3K27me3 but not H3K27me1/H3K27me2 in vitro. We conclude that PHF1 modulates the activity of Ezh2 in favor of the repressive H3K27me3 mark. Thus, we propose that PHF1 is a determinant in PcG-mediated gene repression. PMID- 18285465 TI - DOT1L/KMT4 recruitment and H3K79 methylation are ubiquitously coupled with gene transcription in mammalian cells. AB - The histone H3 lysine 79 methyltransferase DOT1L/KMT4 can promote an oncogenic pattern of gene expression through binding with several MLL fusion partners found in acute leukemia. However, the normal function of DOT1L in mammalian gene regulation is poorly understood. Here we report that DOT1L recruitment is ubiquitously coupled with active transcription in diverse mammalian cell types. DOT1L preferentially occupies the proximal transcribed region of active genes, correlating with enrichment of H3K79 di- and trimethylation. Furthermore, Dot1l mutant fibroblasts lacked H3K79 di- and trimethylation at all sites examined, indicating that DOT1L is the sole enzyme responsible for these marks. Importantly, we identified chromatin immunoprecipitation (ChIP) assay conditions necessary for reliable H3K79 methylation detection. ChIP-chip tiling arrays revealed that levels of all degrees of genic H3K79 methylation correlate with mRNA abundance and dynamically respond to changes in gene activity. Conversion of H3K79 monomethylation into di- and trimethylation correlated with the transition from low- to high-level gene transcription. We also observed enrichment of H3K79 monomethylation at intergenic regions occupied by DNA-binding transcriptional activators. Our findings highlight several similarities between the patterning of H3K4 methylation and that of H3K79 methylation in mammalian chromatin, suggesting a widespread mechanism for parallel or sequential recruitment of DOT1L and MLL to genes in their normal "on" state. PMID- 18285466 TI - Spreading of a corepressor linked to action of long-range repressor hairy. AB - Transcriptional repressor proteins play key roles in the control of gene expression in development. For the Drosophila embryo, the following two functional classes of repressors have been described: short-range repressors such as Knirps that locally inhibit the activity of enhancers and long-range repressors such as Hairy that can dominantly inhibit distal elements. Several long-range repressors interact with Groucho, a conserved corepressor that is homologous to mammalian TLE proteins. Groucho interacts with histone deacetylases and histone proteins, suggesting that it may effect repression by means of chromatin modification; however, it is not known how long-range effects are mediated. Using embryo chromatin immunoprecipitation, we have analyzed a Hairy repressible gene in the embryo during activation and repression. When inactivated, repressors, activators, and coactivators cooccupy the promoter, suggesting that repression is not accomplished by the displacement of activators or coactivators. Strikingly, the Groucho corepressor is found to be recruited to the transcribed region of the gene, contacting a region of several kilobases, concomitant with a loss of histone H3 and H4 acetylation. Groucho has been shown to form higher-order complexes in vitro; thus, our observations suggest that long range effects may be mediated by a "spreading" mechanism, modifying chromatin over extensive regions to inhibit transcription. PMID- 18285467 TI - Blockade of IP3-mediated SK channel signaling in the rat medial prefrontal cortex improves spatial working memory. AB - Planning and directing thought and behavior require the working memory (WM) functions of prefrontal cortex. WM is compromised by stress, which activates phosphatidylinositol (PI)-mediated IP3-PKC intracellular signaling. PKC overactivation impairs WM operations and in vitro studies indicate that IP3 receptor (IP3R)-evoked calcium release results in SK channel-dependent hyperpolarization of prefrontal neurons. However, the effects of IP3R signaling on prefrontal function have not been investigated. The present findings demonstrate that blockade of IP3R or SK channels in the prefrontal cortex enhances WM performance in rats, suggesting that both arms of the PI cascade influence prefrontal cognitive function. PMID- 18285468 TI - Prefrontal cortex and hippocampus subserve different components of working memory in rats. AB - Both the medial prefrontal cortex (mPFC) and hippocampus are implicated in working memory tasks in rodents. Specifically, it has been hypothesized that the mPFC is primarily engaged in the temporary storage and processing of information lasting from a subsecond to several seconds, while the hippocampal function becomes more critical as the working memory demand extends into longer temporal scales. Although these structures may be engaged in a temporally separable manner, the extent of their contributions in the "informational content" of working memory remains unclear. To investigate this issue, the mPFC and dorsal hippocampus (dHPC) were temporarily inactivated via targeted infusions of the GABA(A) receptor agonist muscimol in rats prior to their performance on a delayed alternation task (DAT), employing an automated figure-eight maze that required the animals to make alternating arm choice responses after 3-, 30-, and 60-sec delays for water reward. We report that inactivation of either the mPFC or dHPC significantly reduced DAT at all delay intervals tested. However, there were key qualitative differences in the behavioral effects. Specifically, mPFC inactivation selectively impaired working memory (i.e., arm choice accuracy) without altering reference memory (i.e., the maze task rule) and arm choice response latencies. In contrast, dHPC inactivation increased both reference memory errors and arm choice response latencies. Moreover, dHPC, but not mPFC, inactivation increased the incidence of successive working memory errors. These results suggest that while both the mPFC and hippocampus are necessarily involved in DAT, they seem to process different informational components associated with the memory task. PMID- 18285469 TI - GABA(A) receptors determine the temporal dynamics of memory retention. AB - Four experiments studied the role of GABA(A) receptors in the temporal dynamics of memory retention. Memory for an active avoidance response was a nonmonotonic function of the retention interval. When rats were tested shortly (2 min) or some time (24 h) after training, retention was excellent, but when they were tested at intermediate intervals (1-4 h), retention was poor. Activity at GABA(A) receptors was critical for impairing memory retention at the intermediate intervals because injection of the GABA(A) receptor partial inverse agonist FG7142 prior to test significantly improved performance. These retention enhancing effects of FG7142 were dose-dependent and not due to any nonspecific effects of FG7142 on activity. Our results suggest that the temporal dynamics of memory retention may be caused by variations in neurotransmission through the GABA(A) receptor in the post training period. PMID- 18285470 TI - Activity of daptomycin against Listeria monocytogenes isolates from cerebrospinal fluid. AB - We tested the activity of daptomycin against 76 Listeria monocytogenes isolates from cerebrospinal fluid by broth dilution and Etest methods. For the broth dilution method, the MIC range was 1.0 to 8.0 and the MIC at which 90% of the isolates tested were inhibited (MIC(90)) was 4.0 mg/liter. For the Etest method, the MIC range was 1.0 to 4.0 and the MIC(90) was 4.0 mg/liter. Presently, daptomycin cannot be recommended for the treatment of L. monocytogenes meningitis. PMID- 18285471 TI - Rapid clinical induction of hepatic cytochrome P4502B6 activity by ritonavir. AB - Ritonavir is the most potent and efficacious inhibitor of cytochrome P4503A (CYP3A), and it is used accordingly for the pharmacoenhancement of other antiretrovirals. Paradoxically, ritonavir induces the clinical metabolism and clearance of many drugs. The mechanism by which ritonavir inhibits and induces clinical drug metabolism is unknown. Ritonavir induces CYP2B6 in human hepatocytes. This investigation tested the hypothesis that ritonavir induces human CYP2B6 in vivo. Thirteen healthy human immunodeficiency virus-negative volunteers underwent a three-way sequential crossover protocol, receiving racemic bupropion after nothing (control), 3 days of treatment with ritonavir, and 2.5 weeks of treatment with ritonavir (400 mg twice a day). Stereoselective bupropion hydroxylation was used as an in vivo probe for CYP2B6 activity. Plasma and urine (R)- and (S)-bupropion and (R,R)- and (S,S)-hydroxybupropion concentrations were measured by liquid chromatography-mass spectrometry. Racemic, (R)-, and (S) bupropion plasma ratios of the area under the concentration-time curve from 0 h to infinity (AUC(0-infinity)) (ritonavir/control) were significantly reduced to 0.84, 0.86, and 0.80, respectively, after 3 days of ritonavir treatment and to 0.67, 0.69, and 0.60 after steady-state ritonavir treatment. Apparent oral clearances for racemic, (R)-, and (S)-bupropion all were significantly increased by 1.2-fold after 3 days of ritonavir treatment and by 1.4-, 1.7-, and 1.5-fold after steady-state ritonavir treatment. The plasma (S,S)-hydroxybupropion/(S) bupropion AUC(0-72) ratio was significantly increased by ritonavir. Formation clearances of both (R,R)- and (S,S)-hydroxybupropion were increased 1.8-fold after 3 days of ritonavir treatment and 2.1-fold after steady-state ritonavir treatment. These results show that ritonavir induces human CYP2B6 activity. Induction is rapid, occurring after only 3 days of ritonavir, and is sustained for at least 2 weeks. The ritonavir induction of CYP2B6 activity may have significant implications for drug interactions and clarify previously unexplained interactions. PMID- 18285472 TI - Three thioesterases are involved in the biosynthesis of phosphinothricin tripeptide in Streptomyces viridochromogenes Tu494. AB - Phosphinothricin tripeptide (PTT) is a peptide antibiotic produced by Streptomyces viridochromogenes Tu494, and it is synthesized by nonribosomal peptide synthetases. The PTT biosynthetic gene cluster contains three peptide synthetase genes: phsA, phsB, and phsC. Each of these peptide synthetases comprises only one module. In neither PhsB nor PhsC is a typical C-terminal thioesterase domain present. In contrast, a single thioesterase GXSXG motif has been identified in the N terminus of the first peptide synthetase, PhsA. In addition, two external thioesterase genes, theA and theB, are located within the PTT biosynthetic gene cluster. To analyze the thioesterase function as well as the assembly of the peptide synthetases within PTT biosynthesis, several mutants were generated and analyzed. A phsA deletion mutant (MphsA) was complemented with two different phsA constructs that were carrying mutations in the thioesterase motif. In one construct, the thioesterase motif comprising 45 amino acids of phsA were deleted. In the second construct, the conserved serine residue of the GXSXG motif was replaced by an alanine. In both cases, the complementation of MphsA did not restore PTT biosynthesis, revealing that the thioesterase motif in the N terminus of PhsA is required for PTT production. In contrast, TheA and TheB might have editing functions, as an interruption of the theA and theB genes led to reduced PTT production, whereas an overexpression of both genes in the wild type enhanced the PTT yield. The presence of an active single thioesterase motif in the N terminus of PhsA points to a novel mechanism of product release. PMID- 18285473 TI - Outbreak of Acinetobacter baumannii with chromosomally encoded VIM-1 undetectable by imipenem-EDTA synergy tests. PMID- 18285474 TI - The hepatitis C virus replicon presents a higher barrier to resistance to nucleoside analogs than to nonnucleoside polymerase or protease inhibitors. AB - Specific inhibitors of hepatitis C virus (HCV) replication that target the NS3/4A protease (e.g., VX-950) or the NS5B polymerase (e.g., R1479/R1626, PSI 6130/R7128, NM107/NM283, and HCV-796) have advanced into clinical development. Treatment of patients with VX-950 or HCV-796 rapidly selected for drug-resistant variants after a 14-day monotherapy treatment period. However, no viral resistance was identified after monotherapy with R1626 (prodrug of R1479) or NM283 (prodrug of NM107) after 14 days of monotherapy. Based upon the rapid selection of resistance to the protease and nonnucleoside inhibitors during clinical trials and the lack of selection of resistance to the nucleoside inhibitors, we used the replicon system to determine whether nucleoside inhibitors demonstrate a higher genetic barrier to resistance than protease and nonnucleoside inhibitors. Treatment of replicon cells with nucleoside inhibitors at 10 and 15 times the 50% effective concentration resulted in clearance of the replicon, while treatment with a nonnucleoside or protease inhibitor selected resistant colonies. In combination, the presence of a nucleoside inhibitor reduced the frequency of colonies resistant to the other classes of inhibitors. These results indicate that the HCV replicon presents a higher barrier to the selection of resistance to nucleoside inhibitors than to nonnucleoside or protease inhibitors. Furthermore, the combination of a nonnucleoside or protease inhibitor with a nucleoside polymerase inhibitor could have a clear clinical benefit through the delay of resistance emergence. PMID- 18285475 TI - A peptide with a ProGln C terminus in the human saliva peptidome exerts bactericidal activity against Propionibacterium acnes. AB - Nine proline-rich peptides ending with a proline-glutamine C terminus in a salivary peptidome were sequenced by matrix-assisted laser desorption ionization time of flight time of flight tandem mass spectrometry. A GPPPQGGRPQ peptide binds gram-positive Propionibacterium acnes and considerably inhibits bacterial growth. The peptide exhibiting innate immunity may be applied for treatment of various P. acnes-associated human diseases. PMID- 18285476 TI - Attenuated vancomycin bactericidal activity against Staphylococcus aureus hemB mutants expressing the small-colony-variant phenotype. AB - The in vitro bactericidal activities of vancomycin against Staphylococcus aureus hemB mutants displaying the small-colony-variant phenotype and their parental strains were evaluated. Vancomycin killing activities against hemB mutants were markedly attenuated, demonstrating approximately 50% less effect, a result which was well described by a Hill-type pharmacodynamic model. PMID- 18285477 TI - Effect of low-dose ritonavir on the pharmacokinetics of the CXCR4 antagonist AMD070 in healthy volunteers. AB - AMD070, a CXCR4 antagonist, has demonstrated antiretroviral activity in human immunodeficiency virus-infected patients. Since AMD070 is a substrate of cytochrome P450 3A4 and P-glycoprotein, both of which may be affected by ritonavir, we tested for a ritonavir effect on AMD070 pharmacokinetics. Subjects were given a single 200-mg dose of AMD070 on days 1, 3, and 17. Ritonavir (100 mg every 12 h) was dosed from day 3 to day 18. Blood samples to test for AMD070 concentrations were collected over 48 h after each administration of AMD070. Twenty-three male subjects were recruited. Among them, 21 completed the study, and 2 were discontinued for reasons other than safety. All adverse events were grade 2 or lower. AMD070 alone had the following pharmacokinetic features, given as medians (ranges): 3 h (0.5 to 4 h) for the time to peak blood concentration, 256 ng/ml (41 to 845 ng/ml) for the peak concentration (C(max)), 934 h x ng/ml (313 to 2,127 h x ng/ml) for the area under the concentration-time curve from 0 h to infinity (AUC(0-infinity)), 214 liters/h (94 to 639 liters/h) for apparent body clearance, and 4,201 liters (1,996 to 9,991 liters) for the apparent volume of distribution based on the terminal phase. The initial doses of ritonavir increased the C(max) of AMD070 [geometric mean (90% confidence interval)] by 39% (3 to 89%) and the AUC(0-infinity) by 60% (29 to 100%). After 14 days of ritonavir dosing, the pharmacokinetic changes in AMD070 persisted. The plasma pharmacokinetics of ritonavir were consistent with previous reports. It is concluded that AMD070 concentrations were increased with concomitant ritonavir dosing for 14 days in healthy volunteers. PMID- 18285478 TI - Predictive values of the human immunodeficiency virus phenotype and genotype and of amprenavir and lopinavir inhibitory quotients in heavily pretreated patients on a ritonavir-boosted dual-protease-inhibitor regimen. AB - The inhibitory quotient (IQ) of human immunodeficiency virus (HIV) protease inhibitors (PIs), which is the ratio of drug concentration to viral susceptibility, is considered to be predictive of the virological response. We used several approaches to calculate the IQs of amprenavir and lopinavir in a subset of heavily pretreated patients participating in the French National Agency for AIDS Research (ANRS) 104 trial and then compared their potentials for predicting changes in the plasma HIV RNA level. Thirty-seven patients were randomly assigned to receive either amprenavir (600 mg twice a day [BID]) or lopinavir (400 mg BID) plus ritonavir (100 or 200 mg BID) for 2 weeks before combining the two PIs. The 90% inhibitory concentration (IC(90)) was measured using a recombinant assay without or with additional human serum (IC(90+serum)). Total and unbound PI concentrations in plasma were measured. Univariate linear regression was used to estimate the relation between the change in viral load and the IC(90) or IQ values. The amprenavir phenotypic IQ values were very similar when measured with the standard and protein binding-adjusted IC(90)s. No relationship was found between the viral load decline and the lopinavir IQ. During combination therapy, the amprenavir and lopinavir genotypic IQ values were predictive of the viral response at week 6 (P = 0.03). The number of protease mutations (< 5 or > or = 5) was related to the virological response throughout the study. These findings suggest that the combined genotypic IQ and the number of protease mutations are the best predictors of virological response. High amprenavir and lopinavir concentrations in these patients might explain why plasma concentrations and the phenotypic IQ have poor predictive value. PMID- 18285479 TI - Powerful bactericidal and sterilizing activity of a regimen containing PA-824, moxifloxacin, and pyrazinamide in a murine model of tuberculosis. AB - PA-824 is a nitroimidazo-oxazine in clinical testing for the treatment of tuberculosis. We report that the novel combination of PA-824, moxifloxacin, and pyrazinamide cured mice more rapidly than the first-line regimen of rifampin, isoniazid, and pyrazinamide. If applicable to humans, regimens containing this combination may radically shorten the treatment of multidrug-resistant tuberculosis. PMID- 18285480 TI - A chemical genomic screen in Saccharomyces cerevisiae reveals a role for diphthamidation of translation elongation factor 2 in inhibition of protein synthesis by sordarin. AB - Sordarin and its derivatives are antifungal compounds of potential clinical interest. Despite the highly conserved nature of the fungal and mammalian protein synthesis machineries, sordarin is a selective inhibitor of protein synthesis in fungal organisms. In cells sensitive to sordarin, its mode of action is through preventing the release of translation elongation factor 2 (eEF2) during the translocation step, thus blocking protein synthesis. To further investigate the cellular components required for the effects of sordarin in fungal cells, we have used the haploid deletion collection of Saccharomyces cerevisiae to systematically identify genes whose deletion confers sensitivity or resistance to the compound. Our results indicate that genes in a number of cellular pathways previously unknown to play a role in sordarin response are involved in its growth effects on fungal cells and reveal a specific requirement for the diphthamidation pathway of cells in causing eEF2 to be sensitive to the effects of sordarin on protein synthesis. Our results underscore the importance of the powerful genomic tools developed in yeast (Saccharomyces cerevisiae) to more comprehensively understanding the cellular mechanisms involved in the response to therapeutic agents. PMID- 18285481 TI - Anti-BK virus activity of nucleoside analogs. AB - Polyomavirus BK is an important pathogen in transplant recipients with no effective therapy. This study demonstrates that alkoxyalkyl esters of (S)-9-(3 hydroxy-2-phosphonylmethoxypropyl)adenine and fatty acid derivatives of 9-[2 (phosphonomethyoxy)ethyl]adenine (P393 and P405) are potent and selective inhibitors of BK virus replication in vitro, with a 50% effective concentration in the micromolar-to-nanomolar range. PMID- 18285482 TI - Antimicrobial-resistant pathogens in intensive care units in Canada: results of the Canadian National Intensive Care Unit (CAN-ICU) study, 2005-2006. AB - Between 1 September 2005 and 30 June 2006, 19 medical centers collected 4,180 isolates recovered from clinical specimens from patients in intensive care units (ICUs) in Canada. The 4,180 isolates were collected from 2,292 respiratory specimens (54.8%), 738 blood specimens (17.7%), 581 wound/tissue specimens (13.9%), and 569 urinary specimens (13.6%). The 10 most common organisms isolated from 79.5% of all clinical specimens were methicillin-susceptible Staphylococcus aureus (MSSA) (16.4%), Escherichia coli (12.8%), Pseudomonas aeruginosa (10.0%), Haemophilus influenzae (7.9%), coagulase-negative staphylococci/Staphylococcus epidermidis (6.5%), Enterococcus spp. (6.1%), Streptococcus pneumoniae (5.8%), Klebsiella pneumoniae (5.8%), methicillin-resistant Staphylococcus aureus (MRSA) (4.7%), and Enterobacter cloacae (3.9%). MRSA made up 22.3% (197/884) of all S. aureus isolates (90.9% of MRSA were health care-associated MRSA, and 9.1% were community-associated MRSA), while vancomycin-resistant enterococci (VRE) made up 6.7% (11/255) of all enterococcal isolates (88.2% of VRE had the vanA genotype). Extended-spectrum beta-lactamase (ESBL)-producing E. coli and K. pneumoniae occurred in 3.5% (19/536) and 1.8% (4/224) of isolates, respectively. All 19 ESBL producing E. coli isolates were PCR positive for CTX-M, with bla CTX-M-15 occurring in 74% (14/19) of isolates. For MRSA, no resistance against daptomycin, linezolid, tigecycline, and vancomycin was observed, while the resistance rates to other agents were as follows: clarithromycin, 89.9%; clindamycin, 76.1%; fluoroquinolones, 90.1 to 91.8%; and trimethoprim-sulfamethoxazole, 11.7%. For E. coli, no resistance to amikacin, meropenem, and tigecycline was observed, while resistance rates to other agents were as follows: cefazolin, 20.1%; cefepime, 0.7%; ceftriaxone, 3.7%; gentamicin, 3.0%; fluoroquinolones, 21.1%; piperacillin tazobactam, 1.9%; and trimethoprim-sulfamethoxazole, 24.8%. Resistance rates for P. aeruginosa were as follows: amikacin, 2.6%; cefepime, 10.2%; gentamicin, 15.2%; fluoroquinolones, 23.8 to 25.5%; meropenem, 13.6%; and piperacillin tazobactam, 9.3%. A multidrug-resistant (MDR) phenotype (resistance to three or more of the following drugs: cefepime, piperacillin-tazobactam, meropenem, amikacin or gentamicin, and ciprofloxacin) occurred frequently in P. aeruginosa (12.6%) but uncommonly in E. coli (0.2%), E. cloacae (0.6%), or K. pneumoniae (0%). In conclusion, S. aureus (MSSA and MRSA), E. coli, P. aeruginosa, H. influenzae, Enterococcus spp., S. pneumoniae, and K. pneumoniae are the most common isolates recovered from clinical specimens in Canadian ICUs. A MDR phenotype is common for P. aeruginosa isolates in Canadian ICUs. PMID- 18285483 TI - Prevalence of the erm(T) gene in clinical isolates of erythromycin-resistant group D Streptococcus and Enterococcus. AB - Among 48 erythromycin-resistant group D streptococci (GDS), 36 had the erm(T) resistance gene. erm(T) was also found in 4 of 31 erythromycin-resistant Enterococcus faecium isolates. This is the first report of the erm(T) gene in U.S. GDS isolates and the first report of the erm(T) gene in enterococci. PMID- 18285484 TI - In vivo therapeutic effect of gatifloxacin on BALB/c mice infected with Nocardia brasiliensis. AB - In the present work, we evaluated the effect of gatifloxacin on the evolution of experimental murine infection with Nocardia brasiliensis using linezolid as a control. Gatifloxacin was injected subcutaneously at 100 mg/kg body weight every 8 h for 4 weeks. This compound was equally as efficient as linezolid in reducing the production of lesions. PMID- 18285485 TI - Synergy between polyethylenimine and different families of antibiotics against a resistant clinical isolate of Pseudomonas aeruginosa. AB - The in vitro activities of 10 families of antimicrobial agents alone and in combination with a synthetic polycationic polymer, polyethylenimine (PEI), against a resistant clinical isolate of Pseudomonas aeruginosa were investigated by MIC assays, checkerboard testing, and killing curve studies. At a concentration of 250 nM, PEI (10 kDa) was not directly bactericidal or bacteriostatic; but when it was used in combination with novobiocin, ceftazidime, ampicillin, ticarcillin, carbenicillin, piperacillin, cefotaxime, chloramphenicol, rifampin, or norfloxacin, it significantly reduced the MICs of these antibiotics by 1.5- to 56-fold. However, the MICs of aminoglycosides, polymyxins, and vancomycins were increased by 1.2- to 5-fold when these drugs were combined with PEI; and the MICs of tetracycline, erythromycin, ciprofloxacin, and ofloxacin were not affected when these drugs were combined with PEI. In the killing curve studies, combinations of PEI with novobiocin, ceftazidime, chloramphenicol, or rifampin resulted in 5- to 8-log(10) CFU/ml reductions in bacterial counts when 25% of the MIC of each antibiotic was used. These results indicate that infections due to resistant Pseudomonas strains could be treated by the use of a synergistic combination of PEI and antimicrobial drugs. PMID- 18285486 TI - Prevalence and susceptibility profile of Candida metapsilosis and Candida orthopsilosis: results from population-based surveillance of candidemia in Spain. AB - We describe the prevalences and susceptibility profiles of two recently described species, Candida metapsilosis and Candida orthopsilosis, related to Candida parapsilosis in candidemia. The prevalences of these species (1.7% for C. metapsilosis and 1.4% for C. orthopsilosis) are significant. Differences observed in their susceptibility profiles could have therapeutic importance. PMID- 18285487 TI - Absence of amphotericin B-tolerant persister cells in biofilms of some Candida species. AB - Biofilms and planktonic cells of five Candida species were surveyed for the presence of persister (drug-tolerant) cell populations after exposure to amphotericin B. None of the planktonic cultures (exponential or stationary phase) contained persister cells. However, persisters were found in biofilms of one of two strains of Candida albicans tested and in biofilms of Candida krusei and Candida parapsilosis, but not in biofilms of Candida glabrata or Candida tropicalis. These results suggest that persister cells cannot solely account for drug resistance in Candida biofilms. PMID- 18285488 TI - Plasmid-mediated qepA gene among Escherichia coli clinical isolates from Japan. AB - Seven hundred fifty-one Escherichia coli clinical isolates collected from 140 Japanese hospitals between 2002 and 2006 were screened for the qepA and qnr genes. Two E. coli isolates (0.3%) harbored qepA, but no qnr was identified. The results suggested a low prevalence of E. coli harboring qepA or qnr in Japan. PMID- 18285489 TI - erm(B)-carrying elements in tetracycline-resistant pneumococci and correspondence between Tn1545 and Tn6003. AB - This study investigated the genetic organization of erm(B)-carrying transposons of Streptococcus pneumoniae and their distribution in tetracycline-resistant clinical isolates. By comparatively analyzing reference pneumococci carrying erm(B)/tet(M) transposon Tn1545, Tn6003, Tn6002, or Tn3872, we demonstrated a substantial correspondence between Tn1545 and Tn6003, which have the same resistance gene combination [tet(M) (tetracycline), erm(B) (erythromycin), and aphA-3 (kanamycin)]; share the macrolide-aminoglycoside-streptothricin element, containing a second erm(B); and only differ by a ca. 1.2-kb insertion (containing a putative IS1239 insertion sequence) detected in Tn1545 from S. pneumoniae reference strain BM4200. These results enabled elucidation of the structure of Tn1545, the first erm(B)-carrying transposon described in S. pneumoniae. A collection of 83 erythromycin- and tetracycline-resistant clinical pneumococci, representative of recent Italian isolates carrying erm(B) as the sole erythromycin resistance gene, was used to investigate the distribution of the different transposons. All 83 organisms were positive for tet(M) and bore an erm(B)/tet(M) transposon that could be characterized by using a specific set of primer pairs; Tn3872 was detected in 18 isolates, Tn6002 in 59 isolates, and Tn6003 in 6 (the sole kanamycin-resistant) isolates. The genetic organization of transposon Tn1545, with its specific insertion, was not detected in any of the isolates tested. The erm(B)-carrying elements of tetracycline-resistant pneumococci substantially corresponded to those [bearing a silent tet(M) gene] recently detected in tetracycline-susceptible pneumococci. Overall, in erm(B) positive pneumococci, Tn6003 was the least common erm(B)-carrying Tn916-related element and Tn6002 the most common. PMID- 18285490 TI - LipL32 is an extracellular matrix-interacting protein of Leptospira spp. and Pseudoalteromonas tunicata. AB - LipL32 is the major outer membrane protein in pathogenic Leptospira. It is highly conserved throughout pathogenic species and is expressed in vivo during human infection. While these data suggest a role in pathogenesis, a function for LipL32 has not been defined. Outer membrane proteins of gram-negative bacteria are the first line of molecular interaction with the host, and many have been shown to bind host extracellular matrix (ECM). A search for leptospiral ECM-interacting proteins identified the major outer membrane protein, LipL32. To verify this finding, recombinant LipL32 was expressed in Escherichia coli and was found to bind Matrigel ECM and individual components of ECM, including laminin, collagen I, and collagen V. Likewise, an orthologous protein found in the genome of Pseudoalteromonas tunicata strain D2 was expressed and found to be functionally similar and immunologically cross-reactive. Lastly, binding activity was mapped to the C-terminal 72 amino acids. These studies show that LipL32 and an orthologous protein in P. tunicata are immunologically cross-reactive and function as ECM-interacting proteins via a conserved C-terminal region. PMID- 18285491 TI - Genetic resistance of mice to Mycobacterium paratuberculosis is influenced by Slc11a1 at the early but not at the late stage of infection. AB - We have recently described the development of a luminescent Mycobacterium paratuberculosis strain of bovine origin expressing the luxAB genes of Vibrio harveyi. With this luminescent isolate, fastidious and costly enumeration of CFU by plating them on agar can be replaced by easy and rapid luminometry. Here, we have reevaluated the effect of Slc11a1 (formerly Nramp1) polymorphism on susceptibility to M. paratuberculosis, using this luminometric method. A series of inbred mouse strains were infected intravenously with luminescent M. paratuberculosis S-23 and monitored for bacterial replication in spleen, liver, and lungs for 12 weeks. The results indicate that, as for Mycobacterium avium subsp. avium, innate resistance to infection is genetically controlled by Slc11a1. In BALB/c, congenic BALB.B10-H2(b) (BALB/c background; H-2(b)), C57BL/6, and beige C57BL/6(bg/)(bg) mice (all Slc11a1(s)), bacterial numbers in spleen and liver remained unchanged during the first 4 weeks of infection, whereas in DBA/2 and congenic BALB/c.DBA/2 (C.D2) mice (both Slc11a1(r)) and in (C57BL/6 x DBA/2)F(1) mice (Slc11a1(s/r)), the bacterial numbers had decreased more than 10 fold at 4 weeks postinfection in both male and female mice. At later time points, additional differences in bacterial replication were observed between the susceptible mouse strains, particularly in the liver. Whereas bacterial numbers in the liver gradually decreased more than 100-fold in C57BL/6 mice between week 4 and week 12, bacterial numbers were stable in livers from BALB/c and beige C57BL/6(bg/)(bg) mice during this period. Mycobacterium-specific gamma interferon responses developed earlier and to a higher magnitude in C57BL/6 mice than in BALB/c mice and were lowest in resistant C.D2 mice. PMID- 18285492 TI - Human dendritic cell-specific intercellular adhesion molecule-grabbing nonintegrin (CD209) is a receptor for Yersinia pestis that promotes phagocytosis by dendritic cells. AB - Yersinia pestis is the etiologic agent of bubonic and pneumonic plagues. It is speculated that Y. pestis hijacks antigen-presenting cells (APCs), such as dendritic cells (DCs) and alveolar macrophages, in order to be delivered to lymph nodes. However, how APCs initially capture the bacterium remains uncharacterized. It is well known that HIV-1 uses human DC-specific intercellular adhesion molecule-grabbing nonintegrin (DC-SIGN) (CD209) receptor, expressed by APCs, to be captured and delivered to target cell, such as CD4+ lymphocytes. Several gram negative bacteria utilize their core lipopolysaccharides (LPS) as ligands to interact with the human DC-SIGN. Therefore, it is possible that Y. pestis, whose core LPS is naturally exposed, might exploit DC-SIGN to invade APCs. We demonstrate in this study that Y. pestis directly interacts with DC-SIGN and invades both DCs and alveolar macrophages. In contrast, when engineered to cover the core LPS, Y. pestis loses its ability to invade DCs, alveolar macrophages, and DC-SIGN-expressing transfectants. The interaction between Y. pestis and human DCs can be reduced by a combination treatment with anti-CD209 and anti-CD207 antibodies. This study shows that human DC-SIGN is a receptor for Y. pestis that promotes phagocytosis by DCs in vitro. PMID- 18285493 TI - The EtpA exoprotein of enterotoxigenic Escherichia coli promotes intestinal colonization and is a protective antigen in an experimental model of murine infection. AB - The enterotoxigenic Escherichia coli (ETEC) strains are major causes of morbidity and mortality due to diarrheal illness in developing countries. At present, there is no broadly protective vaccine for this diverse group of pathogens. The EtpA protein, identified in ETEC H10407 in a recent search for candidate immunogens, is a large glycosylated exoprotein secreted via two-partner secretion (TPS). Similar to structurally related molecules, EtpA functions in vitro as an adhesin. The studies reported here use a recently developed murine model of ETEC intestinal colonization to examine the immunogenicity and protective efficacy of EtpA. We report that mice repeatedly exposed to ETEC are protected from subsequent colonization and that they mount immune responses to both EtpA and its presumed two-partner secretion transporter (EtpB) during the course of experimental infection. Furthermore, isogenic etpA deletion mutants were impaired in the colonization of mice, and intranasal immunization of mice with recombinant EtpA conferred protection against ETEC H10407 in this model. Together, these data suggest that EtpA is required for optimal colonization of the intestine, findings paralleling those of previous in vitro studies demonstrating its role in adherence. EtpA and other TPS proteins may be viable targets for ETEC vaccine development. PMID- 18285494 TI - Enhancement of antibody responses to Bacillus anthracis protective antigen domain IV by use of calreticulin as a chimeric molecular adjuvant. AB - The generation of protective humoral immune responses against the receptor binding domain (domain IV) of protective antigen [PA(dIV)] of Bacillus anthracis represents a plausible approach against anthrax toxin. In the current study, we have developed a naked DNA vaccine encoding calreticulin (CRT) linked to PA(dIV) of Bacillus anthracis [CRT/PA(dIV)]. We transfected a human embryonic kidney cell line (HEK 293) with CRT/PA(dIV) DNA and performed Western blotting and confocal microscopy analysis. We found that linkage of CRT to PA(dIV) targets PA(dIV) to the endoplasmic reticulum, resulting in secretion of the chimeric CRT/PA(dIV) protein. We then evaluated the ability of CRT/PA(dIV) DNA to generate PA(dIV) specific antibody responses and protective immunity against lethal anthrax toxin (PA plus lethal factor) challenge. We found that mice immunized with CRT/PA(dIV) DNA were capable of rapidly inducing significantly higher PA(dIV)-specific antibody responses than mice immunized with PA(dIV) DNA alone. Furthermore, we observed that this enhanced antibody response generated by CRT/PA(dIV) DNA was CD4 dependent, since CD4 knockout mice demonstrated a significant reduction in antibody responses. In addition, analysis of the titers and avidity maturation of the induced PA-specific antibodies revealed that vaccination with CRT/PA(dIV) DNA vaccine accelerated the avidity maturation of antibodies to PA(dIV) compared to vaccination with PA(dIV) DNA. Importantly, the enhanced antibody responses correlated to protective immunity against lethal anthrax toxin challenge. Thus, DNA vaccines encoding CRT linked to PA(dIV) may dramatically enhance PA-specific protective antibody responses. Our results have significant clinical applications for biodefense against anthrax toxin. PMID- 18285495 TI - Anaplasma phagocytophilum MSP2(P44)-18 predominates and is modified into multiple isoforms in human myeloid cells. AB - Anaplasma phagocytophilum is the etiologic agent of human granulocytic anaplasmosis. MSP2(P44), the bacterium's major surface protein, is encoded by a paralogous gene family and has been implicated in a variety of pathobiological processes, including antigenic variation, host adaptation, adhesion, porin activity, and structural integrity. The consensus among several studies performed at the DNA and RNA levels is that a heterogeneous mix of a limited number of msp2(p44) transcripts is expressed by A. phagocytophilum during in vitro cultivation. Such analyses have yet to be extended to the protein level. In this study, we used proteomic and molecular approaches to determine that MSP2(P44)-18 is the predominant if not the only paralog expressed and is modified into multiple 42- to 44-kDa isoforms by A. phagocytophilum strain HGE1 during infection of HL-60 cells. The msp2(p44) expression profile was homogeneous for msp2(p44)-18. Thus, MSP2(P44)-18 may have a fitness advantage in HL-60 cell culture in the absence of selective immune pressure. Several novel 22- to 27-kDa MSP2 isoforms lacking most of the N-terminal conserved region were also identified. A. phagocytophilum MSP2(P44) orthologs expressed by other pathogens in the family Anaplasmataceae are glycosylated. Gas chromatography revealed that recombinant MSP2(P44)-18 is modified by glucose, galactose, xylose, mannose, and trace amounts of other glycosyl residues. These data are the first to confirm differential modification of any A. phagocytophilum MSP2(P44) paralog and the first to provide evidence for expression of truncated versions of such proteins. PMID- 18285496 TI - Hepatosplenomegaly is associated with low regulatory and Th2 responses to schistosome antigens in childhood schistosomiasis and malaria coinfection. AB - Hepatosplenomegaly among Kenyan schoolchildren has been shown to be exacerbated where there is transmission of both Schistosoma mansoni and Plasmodium falciparum. This highly prevalent and chronic morbidity often occurs in the absence of ultrasound-detectable periportal fibrosis and may be due to immunological inflammation. For a cohort of school-age children, whole-blood cultures were stimulated with S. mansoni soluble egg antigen (SEA) or soluble worm antigen (SWA). Responses to SWA were found to be predominantly Th2 cytokines; however, they were not significantly associated with either hepatosplenomegaly or infection with S. mansoni or P. falciparum. In comparison, SEA-specific Th2 cytokine responses were low, and the levels were negatively correlated with S. mansoni infection intensities and were lower among children who were coinfected with P. falciparum. Tumor necrosis factor alpha levels in response to stimulation with SEA were high, and a negative association between presentation with hepatomegaly and the levels of the regulatory cytokines interleukin-6 and transforming growth factor beta(1) suggests that a possible mechanism for childhood hepatomegaly in areas where both malaria and schistosomiasis are endemic is poor regulation of an inflammatory response to schistosome eggs. PMID- 18285497 TI - Genetic susceptibility to chronic hepatitis is inherited codominantly in Helicobacter hepaticus-infected AB6F1 and B6AF1 hybrid male mice, and progression to hepatocellular carcinoma is linked to hepatic expression of lipogenic genes and immune function-associated networks. AB - Helicobacter hepaticus causes hepatitis in susceptible strains of mice. Previous studies indicated that A/JCr mice are susceptible and C57BL/6NCr mice are resistant to H. hepaticus-induced hepatitis. We used F1 hybrid mice derived from A/J and C57BL/6 matings to investigate their phenotype and determine their hepatic gene expression profile in response to H. hepaticus infection. F1 hybrid mice, as well as parental A/J and C57BL/6 mice, were divided equally into control and H. hepaticus-infected groups and euthanized at 18 months postinoculation. Hepatic lesions were evaluated histologically and the differential hepatic gene expression in F1 mice was determined by microarray-based global gene expression profiling analysis. H. hepaticus-infected parental strains including A/J and C57BL/6 mice, as well as F1 mice, developed significant hepatitis. Overall, hepatocellular carcinomas or dysplastic liver lesions were observed in 69% of H. hepaticus-infected F1 male mice and H. hepaticus was isolated from hepatic tissues of all F1 mice with liver tumors. Liver tumors, characterized by hepatic steatosis, developed in livers with high hepatitis scores. To identify gene expression specific to H. hepaticus-induced hepatitis and progression to hepatocellular carcinoma in F1 mice, a method using comparative group transcriptome analysis was utilized. The canonical pathway most significantly enriched was immunological disease. Fatty acid synthase and steaoryl-coenzyme A desaturase, the two rate-limiting enzymes in lipogenesis, were upregulated in neoplastic relative to dysplastic livers. This study suggests a synergistic interaction between hepatic steatosis and infectious hepatitis leading to hepatocellular carcinoma. The use of AB6F1 and B6AF1 mice, as well as genetically engineered mice, on a C57BL/6 background will allow studies investigating the role of chronic microbial hepatitis and steatohepatitis in the pathogenesis of liver cancer. PMID- 18285498 TI - Intracellular neutralization of shiga toxin 2 by an a subunit-specific human monoclonal antibody. AB - Infection of children with Shiga toxin (Stx)-producing Escherichia coli (STEC) is the leading cause of hemolytic-uremic syndrome (HUS). Stx2, one of two toxins liberated by the bacteria, is directly linked with HUS. We have previously shown that Stx2-specific human monoclonal antibodies (HuMAbs) protect mice and piglets from fatal systemic complications of Stx2. The present study investigates the mechanisms by which our most efficacious A- and B-subunit-specific HuMAbs neutralize the cytotoxic effects of Stx2 in vitro. Whereas the B-subunit-specific HuMAb 5H8 blocked binding of Stx2 to its receptor on the cell surface, the A subunit-specific HuMAb 5C12 did not interfere with the toxin-receptor binding. Further investigations revealed that 5C12 did not block endocytosis of Stx2 by HeLa cells as both Stx2 and 5C12 colocalized with early endosomes. However, 5C12 blocked the retrograde transport of the toxin into the Golgi and the endoplasmic reticulum, preventing the toxin from entering the cytosol where the toxin exerts its cytotoxic effect. The endocytosed 5C12/Stx2 complexes appear to be rapidly transported to the plasma membrane and/or to the slow recycling perinuclear compartments, followed by their slow recycling to the plasma membrane, and release into the extracellular environment. PMID- 18285499 TI - Pseudomonas aeruginosa cystic fibrosis isolates from individual patients demonstrate a range of levels of lethality in two Drosophila melanogaster infection models. AB - Recently, two Drosophila melanogaster models of infection, fly feeding and fly nicking, have been developed that allow a determination of pathogenic potential of Pseudomonas aeruginosa isolates. In this study, control strains, isolates from burn wounds, and isolates from the sputa of cystic fibrosis (CF) patients were used to compare the two infection models to determine whether any of the isolates might be better adapted to either of the models. In addition, our goal was to determine the variability of isolates from individual CF patients. Three of four control strains (PAO1, PAK, and PA14) caused significant mortality in the flies in both models of infection. The remaining control strain, PA103, was lethal to flies in the nicking model but lacked significant lethality in the feeding model. The burn wound isolates had a high level of lethality in both models. Interestingly, the CF isolates had the largest diversity of lethality in both models of infection. The range of pathogenic potentials of the CF isolates occurred across a cohort of patients, both at the patient level and down to the level of individual sputum samples. The majority of all isolates had similar levels of lethality in both fly infection models. However, two CF isolates were significantly more lethal in the nicking model, and three CF isolates were significantly more lethal in the feeding model. In conclusion, the two Drosophila infection models were useful for the analysis of the diversity of pathogenic potentials of P. aeruginosa isolates. PMID- 18285500 TI - O antigen protects Bordetella parapertussis from complement. AB - Bordetella pertussis, a causative agent of whooping cough, expresses BrkA, which confers serum resistance, but the closely related human pathogen that also causes whooping cough, Bordetella parapertussis, does not. Interestingly, B. parapertussis, but not B. pertussis, produces an O antigen, a factor shown in other models to confer serum resistance. Using a murine model of infection, we determined that O antigen contributes to the ability of B. parapertussis to colonize the respiratory tract during the first week of infection, but not thereafter. Interestingly, an O antigen-deficient strain of B. parapertussis was not defective in colonizing mice lacking the complement cascade. O antigen prevented both complement component C3 deposition on the surface and complement mediated killing of B. parapertussis. In addition, O antigen was required for B. parapertussis to systemically spread in complement-sufficient mice, but not complement-deficient mice. These data indicate that O antigen enables B. parapertussis to efficiently colonize the lower respiratory tract by protecting against complement-mediated control and clearance. PMID- 18285501 TI - Protective heterologous immunity against fatal ehrlichiosis and lack of protection following homologous challenge. AB - The roles of antibodies and memory T cells in protection against virulent Ehrlichia have not been completely investigated. In this study, we addressed these issues by using murine models of mild and fatal ehrlichiosis caused by related monocytotropic Ehrlichia strains. Mice were primed with either Ehrlichia muris or closely related virulent ehrlichiae transmitted by Ixodes ovatus (IOE) ticks given intraperitoneally or intradermally. All groups were reinfected intraperitoneally, 30 days later, with a lethal high dose of IOE. Priming with E. muris, but not IOE, induced strong CD4+ and CD8+ memory type 1 T-cell responses, Ehrlichia-specific immunoglobulin G (IgG) antibodies, and persistent infection. Compared to IOE-primed mice, subsequent lethal IOE challenge of E. muris-primed mice, resulted in (i) 100% protection against lethal infection, (ii) strong Ehrlichia-specific secondary gamma interferon (IFN-gamma)-producing effector/effector memory CD4+ and CD8+ T-cell responses, (iii) enhanced secondary anti-ehrlichial antibody response, (iv) accelerated bacterial clearance, and (v) the formation of granulomas in the liver and lung. E. muris-primed mice challenged with IOE had lower levels of serum interleukin-1alpha (IL-1alpha), IL 6, and IL-10 compared to unprimed mice challenged with IOE. Interestingly, the fatal secondary response in IOE-primed mice correlated with (i) decline in the Ehrlichia-specific CD4+ and CD8+ type 1 responses, (ii) marked hepatic apoptosis and necrosis, and (iii) substantial bacterial clearance, suggesting that fatal secondary response is due to immune-mediated tissue damage. In conclusion, protection against fatal ehrlichial infection correlates with strong expansion of IFN-gamma-producing CD4+ and CD8+ effector memory type 1 T cells, which appear to be maintained in the presence of IgG antibodies and persistent infection. PMID- 18285504 TI - A mindfulness-based health wellness program for an adolescent with Prader-Willi syndrome. AB - Individuals with Prader-Willi syndrome have hyperphagia, a characteristic eating disorder defined by a marked delay in the satiety response when compared to controls. This eating disorder has been particularly difficult to control. The authors taught and evaluated effectiveness of regular exercise alone, regular exercise plus healthy eating, and mindfulness-based strategies combined with exercise and healthy eating to an adolescent with this syndrome. Mindfulness based strategies included mindful eating, visualizing and labeling hunger, and rapidly shifting attention away from hunger by engaging in Meditation on the Soles of the Feet. On average, when compared to baseline levels, there were decreases in weight with regular exercise and exercise plus healthy eating, but the most consistent and sustained changes were evidenced when mindfulness training was added to exercise and healthy eating. The adolescent continued using the mindfulness health wellness program and further reduced his weight during the 3-year follow-up period. PMID- 18285502 TI - Application of massively parallel sequencing to microRNA profiling and discovery in human embryonic stem cells. AB - MicroRNAs (miRNAs) are emerging as important, albeit poorly characterized, regulators of biological processes. Key to further elucidation of their roles is the generation of more complete lists of their numbers and expression changes in different cell states. Here, we report a new method for surveying the expression of small RNAs, including microRNAs, using Illumina sequencing technology. We also present a set of methods for annotating sequences deriving from known miRNAs, identifying variability in mature miRNA sequences, and identifying sequences belonging to previously unidentified miRNA genes. Application of this approach to RNA from human embryonic stem cells obtained before and after their differentiation into embryoid bodies revealed the sequences and expression levels of 334 known plus 104 novel miRNA genes. One hundred seventy-one known and 23 novel microRNA sequences exhibited significant expression differences between these two developmental states. Owing to the increased number of sequence reads, these libraries represent the deepest miRNA sampling to date, spanning nearly six orders of magnitude of expression. The predicted targets of those miRNAs enriched in either sample shared common features. Included among the high-ranked predicted gene targets are those implicated in differentiation, cell cycle control, programmed cell death, and transcriptional regulation. PMID- 18285505 TI - Examination of a social problem-solving intervention to treat selective mutism. AB - The authors examined the use of a social problem-solving intervention to treat selective mutism with 2 sisters in an elementary school setting. Both girls were taught to answer teacher questions in front of their classroom peers during regular classroom instruction. Each girl received individualized instruction from a therapist and was taught to discriminate salient social cues, select an appropriate social response, perform the response, and evaluate her performance. The girls generalized the skills to their respective regular classrooms and maintained the skills for up to 3 months after the removal of the intervention. Experimental control was demonstrated using a multiple baseline design across participants. Limitations of this study and issues for future research are discussed. PMID- 18285506 TI - A pilot study of sensation-focused intensive treatment for panic disorder with moderate to severe agoraphobia: preliminary outcome and benchmarking data. AB - This report presents results of a treatment for panic disorder with moderate to severe agoraphobia (PDA-MS) called sensation-focused intensive treatment (SFIT). SFIT is an 8-day intensive treatment that combines features of cognitive- behavioral treatment for panic disorder, such as interoceptive exposure and cognitive restructuring with ungraded situational exposure. SFIT focuses on feared physical sensations as well as agoraphobic avoidance. Preliminary data support the utility of SFIT in improving PDA-MS. The goal of this exploratory study was to further investigate the effectiveness of SFIT and evaluate factors related to treatment outcome, including severity of panic symptoms, gender, comorbidity, self-efficacy, and place of residence (local vs. remote). SFIT was found to be effective in decreasing panic symptoms from pre- to posttreatment, with treatment gains maintained at follow-up. The implications of these findings for the treatment of PDA are discussed. PMID- 18285507 TI - Habituation of premonitory sensations during exposure and response prevention treatment in Tourette's syndrome. AB - Exposure to premonitory sensations and response prevention of tics (ER) has been shown to be a promising new treatment for Tourette's syndrome (TS). The present study tested the hypothesis that habituation to unpleasant premonitory sensations associated with the tic is an underlying mechanism of change in ER. Patients rated the severity of sensations and urges at 15-minute intervals during ten 2 hour ER sessions. Multilevel models using multiple time trend analyses showed significant reductions of the sensory severity ratings both within and between sessions. The decrease of these severity ratings was related to the frequency of tics exhibited during sessions, regardless of tic severity at baseline. These results support the hypothesis that habituation may be at least part of the underlying working mechanism of exposure in the treatment of tics in TS and that effective tic suppression during sessions is an important factor in this habituation process. PMID- 18285508 TI - The assessment of basic learning abilities test for predicting learning of persons with intellectual disabilities: a review. AB - The Assessment of Basic Learning Abilities (ABLA) Test uses standard prompting and reinforcement procedures to assess the ease or difficulty with which a testee is able to learn a simple imitation and five two-choice discriminations. The authors review studies that have examined performance of participants with developmental disabilities (DD) on the ABLA test to predict (a) performance on a variety of simple imitations and two-choice discriminations, (b) performance on three-choice and four-choice discriminations, (c) the relative efficacy of three presentation modes (objects vs. photographs vs. verbal descriptions) for assessing preferences, (d) compliance of adults with DD and children with and without DD, and (e) participants' ability to learn to respond to the spoken names of pictures of common objects. Across all five types of studies, the predictive validity of the ABLA test has been very high. PMID- 18285509 TI - Role playing: applications in hostage and crisis negotiation skills training. AB - Role playing has been a mainstay of behavioral assessment for decades. In recent years, however, this analogue strategy has also enjoyed widespread application in the field of law enforcement. Most notably, role-play procedures have become an integral component of assessment and training efforts in hostage and crisis negotiation, which attempts to resolve high-risk and often volatile situations in a peaceful, nonviolent manner when possible. The purpose of this paper is to (a) describe development and validation of a role-play test specifically geared toward law enforcement negotiators, (b) present different role-play formats that have been incorporated in law enforcement negotiation training, and (c) discuss limitations and considerations in use of these instruments. Suggestions for directions that future efforts in this area might take are offered. The heuristic value of role playing in crisis management, counterterrorism, and emergency and mass casualty disaster training exercises is also underscored. PMID- 18285510 TI - Endogenous regulators of G protein signaling differentially modulate full and partial mu-opioid agonists at adenylyl cyclase as predicted by a collision coupling model. AB - Regulator of G protein signaling (RGS) proteins accelerate the endogenous GTPase activity of Galpha(i/o) proteins to increase the rate of deactivation of active Galpha-GTP and Gbetagamma signaling molecules. Previous studies have suggested that RGS proteins are more effective on less efficiently coupled systems such as with partial agonist responses. To determine the role of endogenous RGS proteins in functional responses to mu-opioid agonists of different intrinsic efficacy, Galpha(i/o) subunits with a mutation at the pertussis toxin (PTX)-sensitive cysteine (C351I) and with or without a mutation at the RGS binding site (G184S) were stably expressed in C6 glioma cells expressing a mu-opioid receptor. Cells were treated overnight with PTX to inactivate endogenous G proteins. Maximal inhibition of forskolin-stimulated adenylyl cyclase by the low-efficacy partial agonists buprenorphine and nalbuphine was increased in cells expressing RGS insensitive Galpha(o)(CIGS), Galpha(i2)(CIGS), or Galpha(i3)(CIGS) compared with their Galpha(CI) counterparts, but the RGS-insensitive mutation had little or no effect on the maximal inhibition by the higher efficacy agonists DAMGO and morphine. The potency of all the agonists to inhibit forskolin-stimulated adenylyl cyclase was increased in cells expressing RGS-insensitive Galpha(o)(CIGS), Galpha(i2)(CIGS), or Galpha(i3)(CIGS), regardless of efficacy. These data are comparable with predictions based on a collision coupling model. In this model, the rate of G protein inactivation, which is modulated by RGS proteins, and the rate of G protein activation, which is affected by agonist intrinsic efficacy, determine the maximal agonist response and potency at adenylyl cyclase under steady state conditions. PMID- 18285511 TI - Chitin signaling in plants: insights into the perception of fungal pathogens and rhizobacterial symbionts. PMID- 18285512 TI - Besides adhesion: new perspectives of integrin functions in angiogenesis. AB - During angiogenic remodelling in embryo and adult life, endothelial cells lining blood vessel walls dynamically modify their integrin-mediated adhesive contacts with the surrounding extracellular matrix. However, besides regulating cell adhesion and migration, integrins dynamically participate in a network with soluble molecules and their receptors. Angiogenesis is characterized by opposing autocrine and paracrine loops of growth factors and semaphorins that regulate the activation of integrins on the endothelial surface through tyrosine kinase receptors (TKR) and the neuropilin/plexin system. Moreover, pro- and anti angiogenic factors can directly bind integrins and regulate endothelial cell behaviour. This review summarizes the recent progress in understanding the reciprocal interactions between integrins, TKR, and semaphorin receptors. PMID- 18285513 TI - Msx1 and Msx2 are functional interacting partners of T-box factors in the regulation of Connexin43. AB - AIMS: T-box factors Tbx2 and Tbx3 play key roles in the development of the cardiac conduction system, atrioventricular canal, and outflow tract of the heart. They regulate the gap-junction-encoding gene Connexin43 (Cx43) and other genes critical for heart development and function. Discovering protein partners of Tbx2 and Tbx3 will shed light on the mechanisms by which these factors regulate these gene programs. METHODS AND RESULTS: Employing an yeast 2-hybrid screen and subsequent in vitro pull-down experiments we demonstrate that muscle segment homeobox genes Msx1 and Msx2 are able to bind the cardiac T-box proteins Tbx2, Tbx3, and Tbx5. This interaction, as that of the related Nkx2.5 protein, is supported by the T-box and homeodomain alone. Overlapping spatiotemporal expression patterns of Msx1 and Msx2 together with the T-box genes during cardiac development in mouse and chicken underscore the biological significance of this interaction. We demonstrate that Msx proteins together with Tbx2 and Tbx3 suppress Cx43 promoter activity and down regulate Cx43 gene activity in a rat heart-derived cell line. Using chromatin immunoprecipitation analysis we demonstrate that Msx1 can bind the Cx43 promoter at a conserved binding site located in close proximity to a previously defined T-box binding site, and that the activity of Msx proteins on this promoter appears dependent in the presence of Tbx3. CONCLUSION: Msx1 and Msx2 can function in concert with the T-box proteins to suppress Cx43 and other working myocardial genes. PMID- 18285514 TI - Angiopoietin-1 prevents hypertension and target organ damage through its interaction with endothelial Tie2 receptor. AB - AIMS: The endothelium has emerged recently as a therapeutic target in the treatment of hypertension because endothelial dysfunction and subsequent vascular rarefaction cause target organ damage and further elevate blood pressure (BP). It led us to hypothesize that one of the endothelial survival factors, a potent derivative of angiopoietin-1 (cartilage oligomeric matrix protein, COMP-Ang-1), could be a novel class of antihypertensive agents that maintain endothelial integrity and function, thereby preventing the development of hypertension and target organ damage. METHODS AND RESULTS: To study the role of COMP-Ang-1 in preventing hypertension and target organ damage, a COMP-Ang-1 plasmid was electroporated into adductor muscles of 6 weeks old, pre-hypertensive, spontaneously hypertensive rats (SHRs), and the secretion of its expressed protein into the bloodstream was confirmed by western blotting. In comparison with sham and reporter gene transfer, COMP-Ang-1 gene transfer significantly prevented increases in systolic BP and reduced microvascular rarefaction and tissue damage in the heart and kidney. However, overexpression of soluble Tie2 receptor completely abolished these beneficial effects of COMP-Ang-1 gene transfer on SHRs, indicating that expressed COMP-Ang-1 protein has antihypertensive effects in SHRs by binding Tie2 receptors on the vascular endothelium. In particular, COMP-Ang-1 gene-transferred SHRs had significantly higher plasma levels of nitrite than other controls, which was found to be due to that expressed COMP-Ang-1 protein promoted nitrite synthesis by activating endothelial nitric oxide synthase, one of the Tie2 downstream-signalling molecules. CONCLUSION: The present study suggests a new potential of endothelial survival factor, COMP-Ang-1, as an antihypertensive agent that effectively reduces the hypertension-associated cardiovascular and renal damage, as well as prevents the further elevation of BP. PMID- 18285516 TI - Cyanide and uncoupling protein function. PMID- 18285517 TI - FANCM of the Fanconi anemia core complex is required for both monoubiquitination and DNA repair. AB - In response to DNA damage, the Fanconi anemia (FA) core complex functions as a signaling machine for monoubiquitination of FANCD2 and FANCI. It remains unclear whether this complex can also participate in subsequent DNA repair. We have shown previously that the FANCM constituent of the complex contains a highly conserved helicase domain and an associated ATP-dependent DNA translocase activity. Here we show that FANCM also possesses an ATP-independent binding activity and an ATP dependent bi-directional branch-point translocation activity on a synthetic four way junction DNA, which mimics intermediates generated during homologous recombination or at stalled replication forks. Using an siRNA-based complementation system, we found that the ATP-dependent activities of FANCM are required for cellular resistance to a DNA-crosslinking drug, mitomycin C, but not for the monoubiquitination of FANCD2 and FANCI. In contrast, monoubiquitination requires the entire helicase domain of FANCM, which has both ATP dependent and independent activities. These data are consistent with participation of FANCM and its associated FA core complex in the FA pathway at both signaling through monoubiquitination and the ensuing DNA repair. PMID- 18285519 TI - Differential expression of signal transduction factors in ovarian follicle development: a functional role for betaglycan and FIBP in granulosa cells in cattle. AB - Ovarian follicles develop in groups yet individual follicles follow different growth trajectories. This growth and development are regulated by endocrine and locally produced growth factors that use a myriad of receptors and signal transduction pathways to exert their effects on theca and granulosa cells. We hypothesize that differential growth may be due to differences in hormonal responsiveness that is partially mediated by differences in expression of genes involved in signal transduction. We used the bovine dominant follicle model, microarrays, quantitative real-time PCR and RNA interference to examine this. We identified 83 genes coding for signal transduction molecules and validated a subset of them associated with different stages of the follicle wave. We suggest important roles for CAM kinase-1 and EphA4 in theca cells and BCAR1 in granulosa cells for the development of dominant follicles and for betaglycan and FIBP in granulosa cells of regressing subordinate follicles. Inhibition of genes for betaglycan and FIBP in granulosa cells in vitro suggests that they inhibit estradiol production in regressing subordinate follicles. PMID- 18285520 TI - HepG2/C3A cells respond to cysteine deprivation by induction of the amino acid deprivation/integrated stress response pathway. AB - To further define genes that are differentially expressed during cysteine deprivation and to evaluate the roles of amino acid deprivation vs. oxidative stress in the response to cysteine deprivation, we assessed gene expression in human hepatoma cells cultured in complete or cysteine-deficient medium. Overall, C3A cells responded to cysteine deprivation by activation of the eukaryotic initiation factor (eIF)2alpha kinase-mediated integrated stress response to inhibit global protein synthesis; increased expression of genes containing amino acid response elements (ASNS, ATF3, CEBPB, SLC7A11, and TRIB3); increased expression of genes for amino acid transporters (SLC7A11, SLC1A4, and SLC3A2), aminoacyl-tRNA synthetases (CARS), and, to a limited extent, amino acid metabolism (ASNS and CTH); increased expression of genes that act to suppress growth (STC2, FOXO3A, GADD45A, LNK, and INHBE); and increased expression of several enzymes that favor glutathione synthesis and maintenance of protein thiol groups (GCLC, GCLM, SLC7A11, and TXNRD1). Although GCLC, GCLM, SLC7A11, HMOX, and TXNRD1 were upregulated, most genes known to be upregulated via oxidative stress were not affected by cysteine deprivation. Because most genes known to be upregulated in response to eIF2alpha phosphorylation and activating transcription factor 4 (ATF4) synthesis were differentially expressed in response to cysteine deprivation, it is likely that many responses to cysteine deprivation are mediated, at least in part, by the general control nondepressible 2 (GCN2)/ATF4 dependent integrated stress response. This conclusion was supported by the observation of similar differential expression of a subset of genes in response to leucine deprivation. A consequence of sulfur amino acid restriction appears to be the upregulation of the cellular capacity to cope with oxidative and chemical stresses via the integrated stress response. PMID- 18285521 TI - Effects of chromosome 17 on features of the metabolic syndrome in the Lyon hypertensive rat. AB - The metabolic syndrome (involving obesity, hypertension, dyslipidemia, insulin resistance, and a proinflammatory/prethrombotic state) is a major risk factor for cardiovascular disease. Its incidence continues to rise, in part because of the epidemic increase in obesity. The Lyon hypertensive (LH) rat is a model for hypertension and several other features of the metabolic syndrome, having high body weight, plasma cholesterol, and triglycerides, increased insulin-to-glucose ratio, and salt-sensitive hypertension. Previous genetic studies in LH/Mav rats and a normotensive control (LN/Mav) identified quantitative trait loci (QTLs) on rat chromosome (RNO)17 for multiple features of the metabolic syndrome. To further evaluate the role of RNO17 in the LH rat, we generated a consomic strain (LH-17(BN)) by substituting LH RNO17 with that of the sequenced Brown Norway (BN/NHsdMcwi) rat. Male LH and BN rats and LH-17(BN) rats were characterized for blood pressure and metabolic and morphological parameters. Similar to the protective effect of LN alleles, the LH-17(BN) rat also showed decreased body weight, triglycerides, and blood pressure; however, there was no significant difference in cholesterol or insulin-to-glucose ratio. Therefore, the substitution of the LH chromosome 17 is sufficient to recapitulate some, but not all, of the traits previously mapped to this chromosome. This could be due to the lack of a susceptible LH genome background or due to the introgression of chromosome 17 from another strain. Regardless, this study provides a single chromosome genetic model for further dissection of blood pressure and morphological and metabolic traits on this chromosome. PMID- 18285522 TI - Familial hypertrophic cardiomyopathy-related cardiac troponin C mutation L29Q affects Ca2+ binding and myofilament contractility. AB - The cardiac troponin C (cTnC) mutation, L29Q, has been found in a patient with familial hypertrophic cardiomyopathy. We previously showed that L29, together with neighboring residues, Asp2, Val28, and Gly30, plays an important role in determining the Ca(2+) affinity of site II, the regulatory site of mammalian cardiac troponin C (McTnC). Here we report on the Ca(2+) binding characteristics of L29Q McTnC and D2N/V28I/L29Q/G30D McTnC (NIQD) utilizing the Phe(27) --> Trp (F27W) substitution, allowing one to monitor Ca(2+) binding and release. We also studied the effect of these mutants on Ca(2+) activation of force generation in single mouse cardiac myocytes using cTnC replacement, together with sarcomere length (SL) dependence. The Ca(2+)-binding affinity of site II of L29Q McTnC(F27W) and NIQD McTnC(F27W) was approximately 1.3- and approximately 1.9 fold higher, respectively, than that of McTnC(F27W). The Ca(2+) disassociation rate from site II of L29Q McTnC(F27W) and NIQD McTnC(F27W) was not significantly different than that of control (McTnC(F27W)). However, the rate of Ca(2+) binding to site II was higher in L29Q McTnC(F27W) and NIQD McTnC(F27W) relative to control (approximately 1.5-fold and approximately 2.0-fold respectively). The Ca(2+) sensitivity of force generation was significantly higher in myocytes reconstituted with L29Q McTnC (approximately 1.4-fold) and NIQD McTnC (approximately 2-fold) compared with those reconstituted with McTnC. Interestingly, the change in Ca(2+) sensitivity of force generation in response to an SL change (1.9, 2.1, and 2.3 mum) was significantly reduced in myocytes containing L29Q McTnC or NIQD McTnC. These results demonstrate that the L29Q mutation enhances the Ca(2+)-binding characteristics of cTnC and that when incorporated into cardiac myocytes, this mutant alters myocyte contractility. PMID- 18285523 TI - Neurocircuits integrating hormone and nutrient signaling in control of glucose metabolism. AB - As obesity, diabetes, and associated comorbidities are on a constant rise, large efforts have been put into better understanding the cellular and molecular mechanisms by which nutrients and metabolic signals influence central and peripheral energy regulation. For decades, peripheral organs as a source and a target of such cues have been the focus of study. Their ability to integrate metabolic signals is essential for balanced energy and glucose metabolism. Only recently has the pivotal role of the central nervous system in the control of fuel partitioning been recognized. The rapidly expanding knowledge on the elucidation of molecular mechanisms and neuronal circuits involved is the focus of this review. PMID- 18285524 TI - Hypothalamic pathways linking energy balance and reproduction. AB - During periods of metabolic stress, animals must channel energy toward survival and away from processes such as reproduction. The reproductive axis, therefore, has the capacity to respond to changing levels of metabolic cues. The cellular and molecular mechanisms that link energy balance and reproduction, as well as the brain sites mediating this function, are still not well understood. This review focuses on the best characterized of the adiposity signals: leptin and insulin. We examine their reproductive role acting on the classic metabolic pathways of the arcuate nucleus, NPY/AgRP and POMC/CART neurons, and the newly identified kisspeptin network. In addition, other hypothalamic nuclei that may play a role in linking metabolic state and reproductive function are discussed. The nature of the interplay between these elements of the metabolic and reproductive systems presents a fascinating puzzle, whose pieces are just beginning to fall into place. PMID- 18285525 TI - Decreased clearance of serum retinol-binding protein and elevated levels of transthyretin in insulin-resistant ob/ob mice. AB - Serum retinol-binding protein (RBP4) is secreted by liver and adipocytes and is implicated in systemic insulin resistance in rodents and humans. RBP4 normally binds to the larger transthyretin (TTR) homotetramer, forming a protein complex that reduces renal clearance of RBP4. To determine whether alterations in RBP4 TTR binding contribute to elevated plasma RBP4 levels in insulin-resistant states, we investigated RBP4-TTR interactions in leptin-deficient ob/ob mice and high-fat-fed obese mice (HFD). Gel filtration chromatography of plasma showed that 88-94% of RBP4 is contained within the RBP4-TTR complex in ob/ob and lean mice. Coimmunoprecipitation with an RBP4 antibody brought down stoichiometrically equal amounts of TTR and RBP4, indicating that TTR was not more saturated with RBP4 in ob/ob mice than in controls. However, plasma TTR levels were elevated approximately fourfold in ob/ob mice vs. controls. RBP4 injected intravenously in lean mice cleared rapidly, whereas the t(1/2) for disappearance was approximately twofold longer in ob/ob plasma. Urinary fractional excretion of RBP4 was reduced in ob/ob mice, consistent with increased retention. In HFD mice, plasma TTR levels and clearance of injected RBP4 were similar to chow-fed controls. Hepatic TTR mRNA levels were elevated approximately twofold in ob/ob but not in HFD mice. Since elevated circulating RBP4 causes insulin resistance and glucose intolerance in mice, these findings suggest that increased TTR or alterations in RBP4-TTR binding may contribute to insulin resistance by stabilizing RBP4 at higher steady state concentrations in circulation. Lowering TTR levels or interfering with RBP4 TTR binding may enhance insulin sensitivity in obesity and type 2 diabetes. PMID- 18285526 TI - Thermogenic effect of triiodothyroacetic acid at low doses in rat adipose tissue without adverse side effects in the thyroid axis. AB - Triiodothyroacetic acid (TRIAC) is a physiological product of triiodothyronine (T(3)) metabolism, with high affinity for T(3) nuclear receptors. Its interest stems from its potential thermogenic effects. Thus this work aimed 1) to clarify these thermogenic effects mediated by TRIAC vs. T(3) in vivo and 2) to determine whether they occurred predominantly in adipose tissues. To examine this, control rats were infused with equimolar T(3) or TRIAC doses (0.8 or 4 nmolx100 g body wt(-1) x day(-1)) or exposed for 48 h to cold. Both T(3) doses and only the highest TRIAC dose inhibited plasma and pituitary thyroid-stimulating hormone (TSH) and thyroxine (T(4)) in plasma and tissues. Interestingly, the lower TRIAC dose marginally inhibited plasma T(4). T(3) infusion increased plasma and tissue T(3) in a tissue-specific manner. The highest TRIAC dose increased TRIAC concentrations in plasma and tissues, decreasing plasma T(3). TRIAC concentrations in tissues were <10% those of T(3). Under cold exposure or high T(3) doses, TRIAC increased only in white adipose tissue (WAT). Remarkably, only the lower TRIAC dose activated thermogenesis, inducing ectopic uncoupling protein (UCP)-1 expression in WAT and maximal increases in UCP-1, UCP-2, and lipoprotein lipase (LPL) expression in brown adipose tissue (BAT), inhibiting UCP-2 in muscle and LPL in WAT. TRIAC, T(3), and cold exposure inhibited leptin secretion and mRNA in WAT. In summary, TRIAC, at low doses, induces thermogenic effects in adipose tissues without concomitant inhibition of TSH or hypothyroxinemia, suggesting a specific role regulating energy balance. This selective effect of TRIAC in adipose tissues might be considered a potential tool to increase energy metabolism. PMID- 18285527 TI - PYY3-36 injection in mice produces an acute anorexigenic effect followed by a delayed orexigenic effect not observed with other anorexigenic gut hormones. AB - Peptide YY (PYY) is secreted postprandially from the endocrine L cells of the gastrointestinal tract. PYY(3-36), the major circulating form of the peptide, is thought to reduce food intake in humans and rodents via high-affinity binding to the autoinhibitory neuropeptide Y (NPY) receptor within the arcuate nucleus. We studied the effect of early light-phase injection of PYY(3-36) on food intake in mice fasted for 0, 6, 12, 18, 24, and 30 h and show that PYY(3-36) produces an acute anorexigenic effect regardless of the duration of fasting. We also show evidence of a delayed orexigenic effect in ad libitum-fed mice injected with PYY(3-36) in the early light phase. This delayed orexigenic effect also occurs in mice administered a potent analog of PYY(3-36), d-Allo Ile(3) PYY(3-36), but not following injection of other anorectic agents (glucagon-like-peptide 1, oxyntomodulin, and lithium chloride). Early light-phase injection of PYY(3-36) to ad libitum-fed mice resulted in a trend toward increased levels of hypothalamic NPY and agouti-related peptide mRNA and a decrease in proopiomelanocortin mRNA at the beginning of the dark phase. Furthermore, plasma levels of ghrelin were increased significantly, and there was a trend toward decreased plasma PYY(3-36) levels at the beginning of the dark phase. These data indicate that PYY(3-36) injection results in an acute anorexigenic effect followed by a delayed orexigenic effect. PMID- 18285528 TI - Effects of a high-glucose environment on the pituitary growth hormone-releasing hormone receptor: type 1 diabetes compared with in vitro glucotoxicity. AB - The present study investigated the effects of diabetes and high glucose on GHRH receptor (GHRH-R) mRNA and protein levels in the pituitary of diabetic rats 2, 21, and 60 days post-streptozotocin (post-STZ) administration. Two days post-STZ, the 2.5-kb GHRH-R mRNA transcript was increased. Twenty-one days post-STZ, both the 2.5- and 4-kb transcripts and a 72-kDa (125)I-GHRH-GHRH-R complex were elevated. Sixty days post-STZ, the 4-kb transcript remained increased and the 45 kDa (125)I-GHRH-GHRH-R complex (functional receptor) was decreased. Hypothalamic GHRH mRNA and serum total IGF-I levels were reduced at all three time points. To better understand the role of high glucose on GHRH-R regulation, time-course effects of 33 compared with 6 mM d-glucose (DG) were examined in cultured anterior pituitary cells from 2-mo-old healthy rats. Membrane lipoperoxidation was present in 33 mM DG, and GHRH-R mRNA levels were diminished after 24 h, Fluo GHRH internalization was marginal after 16-24 h, and GHRH-induced cAMP levels were decreased after 24 and 48 h. Altogether, these results indicate that the increase of the 2.5-kb GHRH-R mRNA transcript in vivo could be a consequence of a decrease of hypothalamic GHRH mRNA levels in STZ rats. Since it does not affect primarily functional GHRH-R levels, the initial diminution of circulating IGF-I levels could result from a decreased GHRH-R stimulation by GHRH. Thus, the effect of glucotoxicity would be related to a decrease of functional GHRH-R protein, as observed in rats 60 days post-STZ and in cultured pituitary cells from healthy rats exposed to a high-glucose environment. PMID- 18285529 TI - Understanding drug-induced parkinsonism: separating pearls from oy-sters. PMID- 18285530 TI - Teaching NeuroImage: Wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) from midbrain infarction. PMID- 18285532 TI - Toward a plasma marker for Alzheimer disease: some progress, but still a long way to go. PMID- 18285533 TI - Hypothalamic hamartomas and hedgehogs: not a laughing matter. PMID- 18285534 TI - Genetic markers predictive of chemosensitivity and outcome in gliomatosis cerebri. AB - BACKGROUND: Up-front temozolomide (TMZ) has been recently proposed as a treatment for gliomatosis cerebri (GC), but no predictive or prognostic markers have been identified so far. Because 1p19q codeletion and methylguanine methyl transferase promoter (MGMTP) methylation have been correlated with chemosensitivity of gliomas, their value was investigated in a cohort of patients with GC treated with TMZ. METHODS: A cohort of 25 GC patients who were treated with TMZ was investigated for 1p19q codeletion and O6-methylguanine DNA. RESULTS: Patients with a 1p/19q codeletion had a higher response rate (88% [8/9] vs 25% [4/16], p = 0.002), higher progression-free survival (24.5 vs 13.7 months, p = 0.017), and higher overall survival (66.8 vs 15.2 months, p = 0.011) than patients without 1p/19q codeletion. Fourteen of 19 evaluable tumors for MGMTP status were methylated. MGMTP methylation was associated with 1p/19q codeletion (p = 0.045). Patients with unmethylated MGMTP tended to have a shorter progression-free survival and a higher rate of progressive disease. CONCLUSION: Response rate to temozolomide and prognosis seem tightly correlated to 1p19q loss. The impact of methylguanine methyl transferase promoter methylation status on gliomatosis cerebri is still unsettled in this population. PMID- 18285535 TI - Levetiracetam for the treatment of idiopathic generalized epilepsy with myoclonic seizures. AB - BACKGROUND: Currently, there are no published randomized controlled trials evaluating the efficacy and safety of adjunctive antiepileptic therapy in idiopathic generalized epilepsy with myoclonic seizures. METHODS: This randomized, double-blind, placebo-controlled multicenter trial assessed the efficacy and tolerability of adjunctive treatment with levetiracetam 3,000 mg/day in adolescents (>or=12 years) and adults (or=8 days during a prospective 8-week baseline period, despite antiepileptic monotherapy. The 8-week baseline period was followed by 4-week up-titration, 12-week evaluation, and 6 week down-titration/conversion periods. RESULTS: Of 122 patients randomized, 120 (levetiracetam, n = 60; placebo, n = 60) were evaluable. Diagnoses were either juvenile myoclonic epilepsy (93.4%) or juvenile absence epilepsy (6.6%). A reduction of >or=50% in the number of days/week with myoclonic seizures was seen in 58.3% of patients in the levetiracetam group and in 23.3% of patients in the placebo group (p < 0.001) during the treatment period. Levetiracetam-treated patients were more likely to respond to treatment than patients receiving placebo (OR = 4.77; 95% CI, 2.12 to 10.77; p < 0.001). Levetiracetam-treated patients had higher freedom from myoclonic seizures (25.0% vs 5.0%; p = 0.004) and all seizure types (21.7% vs 1.7%; p < 0.001) during the evaluation period. The only adverse events more frequent with levetiracetam were somnolence and neck pain. CONCLUSION: These results suggest that levetiracetam is an effective and well tolerated adjunctive treatment for patients with previously uncontrolled idiopathic generalized epilepsy with myoclonic seizures. PMID- 18285536 TI - Molecular pathology and enzyme processing in various phenotypes of acid maltase deficiency. AB - OBJECTIVES: To examine at molecular, biochemical, and muscle pathology level the striking clinical heterogeneity resulting from acid alpha-glucosidase deficiency. METHODS: We investigated 23 patients with infantile-onset or late-onset glycogen storage disease type II by enzyme activity, protein expression by immunoblotting, GAA gene mutations, and muscle pathology including immunolabeling for Golgi and sarcolemmal proteins. RESULTS: The enzyme activity was absent or minimal in infantile-onset cases and variably reduced in late-onset patients. Genotype phenotype correlation (seven novel mutations were found) showed that most late onset patients had the heterozygous IVS1 leaky splicing mutation (one patient was homozygous), but the course of the disease was often difficult to predict on the basis of the mutations alone. All patients showed an abnormal pattern of enzyme protein processing, with increased amounts of the inactive forms and very low or absent amounts of the mature forms. The molecular weight of the mature and the intermediate forms appeared higher in patients' samples than in the control muscle. We observed a Golgi proliferation in muscle fibers possibly caused by the retention of inactive forms of enzyme protein that cannot be correctly targeted from Golgi to lysosomes. The vacuolar membranes expressed sarcolemmal proteins in late-onset but not in infantile-onset patients, suggesting an extensive autophagy and vacuolar membrane remodeling in late-onset patients. CONCLUSIONS: The different protein molecular weight between patients and controls could be due to an excessive sialylation of mutant enzyme: this might be possibly caused by a delayed transport and longer transit of the inactive proteins in the Golgi apparatus. PMID- 18285537 TI - Thrombophilia differences in cerebral venous sinus and lower extremity deep venous thrombosis. AB - OBJECTIVE: To characterize differences in the prevalence of thrombophilic variables in a large cohort of patients with cerebral venous sinus thrombosis (CVST) and lower extremity deep vein thrombosis (DVT). METHODS: An inception cohort of individuals was identified with first lifetime incident CVST between 1995 and 2005 for whom comprehensive thrombophilia testing was available. To test the hypothesis that thrombophilia prevalence differs with respect to thrombus location, test results were compared to a randomly selected group of patients with lower extremity DVT with comprehensive thrombophilia testing. RESULTS: During this time period, 163 patients with CVST were identified who underwent comprehensive thrombophilia testing. Thrombophilia results were abnormal in 29% including anticardiolipin antibodies (17%), heterozygous factor V Leiden (10%), and heterozygous prothrombin G20210A mutation (n = 14/122; 11%). The prothrombin mutation was more than twice as common in patients with CVST (p = 0.04). Activated protein C resistance, factor V Leiden, and protein C deficiency were more common in patients with DVT (p < 0.05 for each comparison). The anticardiolipin antibodies in patients with CVST were primarily low titer IgM isotype. CONCLUSION: The prevalence of selected thrombophilia factors differs comparing patients with cerebral venous sinus thrombosis and deep vein thrombosis. These differences may offer insights into mechanisms governing the geographic distribution of venous thrombosis. PMID- 18285538 TI - Long-term follow-up of idiopathic intracranial hypertension: the Iowa experience. AB - OBJECTIVE: To evaluate recurrent or delayed worsening of papilledema and visual function in patients with idiopathic intracranial hypertension (IIH) followed for more than 10 years. METHODS: This is an Institutional Review Board approved retrospective chart review of 410 patients with the diagnosis of IIH evaluated at the University of Iowa Hospitals and Clinics from January 1984 to January 1996. Of the 410 patients, 20 patients with IIH who were followed over 10 years at the neuro-ophthalmology clinic met the inclusion criteria. Three neuro ophthalmologists independently evaluated and graded the visual field examinations and optic disc stereo-photographs for each follow-up visit (median = 15). RESULTS: Of the 20 patients, 11 demonstrated a stable course of disease without worsening in papilledema or visual field, and 9 patients worsened after a stable course. Of these 9 patients, 6 patients experienced delayed worsening (range: 28 to 135 months from presentation) and 3 patients had recurrence after resolution of papilledema 12 to 78 months from initial resolution of the IIH. CONCLUSION: Idiopathic intracranial hypertension is a chronic condition that may worsen after a period of stability, warranting long-term follow-up. PMID- 18285539 TI - Endovascular treatment of idiopathic intracranial hypertension: clinical and radiologic outcome of 10 consecutive patients. AB - OBJECTIVE: To explore the relation between venous disease and idiopathic intracranial hypertension. BACKGROUND: Optic nerve sheath fenestration and ventricular shunting are the classic methods when medical treatment has failed. Idiopathic intracranial hypertension is caused by venous sinus obstruction in an unknown percentage of cases. Recently, endoluminal venous sinus stenting was proposed as an alternative treatment. METHODS: Ten consecutive patients with refractory idiopathic intracranial hypertension underwent examination with direct retrograde cerebral venography and manometry to characterize the morphologic features and venous pressures in their cerebral venous sinus. All patients demonstrated morphologic obstruction of the venous lateral sinuses. The CSF pressure was measured in all patients. The CSF pressure on lumbar puncture ranged from 27 to 45 mm Hg with normal composition. All patients had headache, and visual acuity loss was noted in eight patients. Funduscopic examination demonstrated papilledema for all patients. All patients had stenting of the venous sinuses. Intrasinus pressures were recorded before and after the procedure and correlated with clinical outcome. RESULTS: Intrasinus pressures were invariably reduced by stenting. For headache, six patients were rendered asymptomatic, two were improved, and two were unchanged after venous sinus stenting for a mean (+/- SD) follow-up of 17 +/- 10.1 months (range 6 to 36 months). Papilledema disappeared in all patients. In all cases, CSF pressure was normalized at 3-month follow-up. In all patients, direct retrograde cerebral venography or multidetector row CT angiography was performed at 6-month follow-up and demonstrated the absence of stent thrombosis. CONCLUSION: The importance of venous sinus disease in the etiology of idiopathic intracranial hypertension is probably underestimated. Patients with idiopathic intracranial hypertension in whom a venous sinus stenosis is demonstrated by a noninvasive radiologic workup should be evaluated with direct retrograde cerebral venography and manometry. In patients with a lesion of the venous sinuses who experienced medical treatment failure, endovascular stent placement seems to be an interesting alternative to classic surgical approaches. PMID- 18285540 TI - TRP channels: functions and involvement in neurologic disease. PMID- 18285541 TI - Rostral midbrain infarction producing isolated lateropulsion. PMID- 18285542 TI - Re: Hospital volume and stroke outcome: does it matter? PMID- 18285543 TI - Re: Migraine in adolescents: association with socioeconomic status and family history. PMID- 18285544 TI - Re: Activities of daily living in frontotemporal dementia and Alzheimer disease. PMID- 18285545 TI - Risk-adjusted therapy of acute lymphoblastic leukemia can decrease treatment burden and improve survival: treatment results of 2169 unselected pediatric and adolescent patients enrolled in the trial ALL-BFM 95. AB - The trial ALL-BFM 95 for treatment of childhood acute lymphoblastic leukemia was designed to reduce acute and long-term toxicity in selected patient groups with favorable prognosis and to improve outcome in poor-risk groups by treatment intensification. These aims were pursued through a stratification strategy using white blood cell count, age, immunophenotype, treatment response, and unfavorable genetic aberrations providing an excellent discrimination of risk groups. Estimated 6-year event-free survival (6y-pEFS) for all 2169 patients was 79.6% (+/- 0.9%). The large standard-risk (SR) group (35% of patients) achieved an excellent 6y-EFS of 89.5% (+/- 1.1%) despite significant reduction of anthracyclines. In the medium-risk (MR) group (53% of patients), 6y-pEFS was 79.7% (+/- 1.2%); no improvement was accomplished by the randomized use of additional intermediate-dose cytarabine after consolidation. Omission of preventive cranial irradiation in non-T-ALL MR patients was possible without significant reduction of EFS, although the incidence of central nervous system relapses increased. In the high-risk (HR) group (12% of patients), intensification of consolidation/reinduction treatment led to considerable improvement over the previous ALL-BFM trials yielding a 6y-pEFS of 49.2% (+/- 3.2%). Compared without previous trial ALL-BFM 90, consistently favorable results in non-HR patients were achieved with significant treatment reduction in the majority of these patients. PMID- 18285546 TI - A PAI-1 (SERPINE1) polymorphism predicts osteonecrosis in children with acute lymphoblastic leukemia: a report from the Children's Oncology Group. AB - As glucocorticoid use increased in acute lymphoblastic leukemia, osteonecrosis became an increasingly frequent complication. Besides increased age, host risk factors are poorly defined. We tested whether 12 polymorphisms were associated with osteonecrosis among patients 10 years and older treated on the CCG1882 protocol. Candidate genes (TYMS, MTHFR, ABCB1, BGLAP, ACP5, LRP5, ESR1, PAI-1, VDR, PTH, and PTHR) were chosen based on putative mechanisms underlying osteonecrosis risk. All children received dexamethasone, with doses varying by treatment arm. A PAI-1 polymorphism (rs6092) was associated with risk of osteonecrosis in univariate (P = .002; odds ratio = 2.79) and multivariate (P = .002; odds ratio = 2.89) analyses (adjusting for gender, age, and treatment arm). Overall, 21 of 78 (26.9%) children with PAI-1 GA/AA genotypes, versus 25 of 214 (11.7%) children with GG genotype, developed osteonecrosis. PAI-1 polymorphisms and PAI-1 serum levels have previously been associated with thrombosis. We conclude that PAI-1 genetic variation may contribute to risk of osteonecrosis. PMID- 18285547 TI - Effects of donor T-cell trafficking and priming site on graft-versus-host disease induction by naive and memory phenotype CD4 T cells. AB - Graft-versus-host disease (GVHD) remains a major cause of morbidity and mortality in allogeneic stem cell transplantation. Effector memory T cells (T(EM)) do not cause GVHD but engraft and mount immune responses, including graft-versus-tumor effects. One potential explanation for the inability of T(EM) to cause GVHD is that T(EM) lack CD62L and CCR7, which are instrumental in directing naive T cells (T(N)) to lymph nodes (LN) and Peyer patches (PP), putative sites of GVHD initiation. Thus T(EM) should be relatively excluded from LN and PP, possibly explaining their inability to cause GVHD. We tested this hypothesis using T cells deficient in CD62L or CCR7, transplant recipients lacking PNAd ligands for CD62L, and recipients without LN and PP or LN, PP, and spleen. Surprisingly, CD62L and CCR7 were not required for T(N)-mediated GVHD. Moreover, in multiple strain pairings, GVHD developed in recipients that lacked LN and PP. Mild GVHD could even be induced in mice lacking all major secondary lymphoid tissues (SLT). Conversely, enforced constitutive expression of CD62L on T(EM) did not endow them with the ability to cause GVHD. Taken together, these data argue against the hypothesis that T(EM) fail to induce GVHD because of inefficient trafficking to LN and PP. PMID- 18285548 TI - Maribavir prophylaxis for prevention of cytomegalovirus infection in allogeneic stem cell transplant recipients: a multicenter, randomized, double-blind, placebo controlled, dose-ranging study. AB - The anti-cytomegalovirus (CMV) activity and safety of oral maribavir in CMV seropositive allogeneic stem-cell transplant recipients were evaluated in a randomized, double-blind, placebo-controlled, dose-ranging study. After engraftment, 111 patients were randomized to receive CMV prophylaxis with maribavir (100 mg twice daily, 400 mg once daily, or 400 mg twice daily) or placebo. Within the first 100 days after transplantation, the incidence of CMV infection based on CMV pp65 antigenemia was lower in each of the respective maribavir groups (15%, P = .046; 19%, P = .116; 15%, P = .053) compared with placebo (39%). Similarly, the incidence of CMV infection based on plasma CMV DNA was lower in each of the respective maribavir groups (7%, P = .001; 11%, P = .007; 19%, P = .038) compared with placebo (46%). Anti-CMV therapy was also used less often in patients receiving each respective dose of maribavir (15%, P = .001; 30%, P = .051; 15%, P = .002) compared with placebo (57%). There were 3 cases of CMV disease in placebo patients but none in the maribavir patients. Adverse events, mostly taste disturbance, nausea, and vomiting, were more frequent with maribavir. Maribavir had no adverse effect on neutrophil or platelet counts. These results show that maribavir can reduce the incidence of CMV infection and, unlike ganciclovir, does not cause myelosuppression. PMID- 18285549 TI - Circulating surfactant protein A (SP-A), a marker of lung injury, is associated with insulin resistance. AB - OBJECTIVES: Impaired lung function and inflammation have both attracted interest as potentially novel risk factors for glucose intolerance, insulin resistance, and type 2 diabetes. We hypothesized that circulating levels of surfactant protein (SP)-A, which reflects interstitial lung injury, could be associated with altered glucose tolerance and insulin resistance. RESEARCH DESIGN AND METHODS: Circulating SP-A concentration and metabolic variables (including insulin sensitivity by minimal model method, n = 89) were measured in 164 nonsmoking men. RESULTS: Circulating SP-A concentration was significantly higher among patients with glucose intolerance and type 2 diabetes than in subjects with normal glucose tolerance, even after adjustment for BMI, age, and smoking status (ex/never). The most significant differences were found in overweight and obese subjects with altered glucose tolerance (n = 59) who showed significantly increased serum SP-A concentrations (by a mean of 24%) compared with obese subjects with normal glucose tolerance (n = 58) (log SP-A 1.54 +/- 0.13 vs. 1.44 +/- 0.13; P < 0.0001). Insulin sensitivity (P = 0.003) contributed independently to 22% of SP-A variance among all subjects. In subjects with altered glucose tolerance, insulin sensitivity (P = 0.01) and fasting triglycerides (P = 0.02) contributed to 37% of SP-A variance. Controlling for serum creatinine or C-reactive protein in these models did not significantly change the results. CONCLUSIONS: Lung-derived SP-A protein was associated with altered glucose tolerance and insulin resistance in 164 nonsmoking men. PMID- 18285550 TI - Acute complications and drug misuse are important causes of death for children and young adults with type 1 diabetes: results from the Yorkshire Register of diabetes in children and young adults. AB - OBJECTIVE: To examine mortality rates and causes of death among subjects diagnosed with type 1 diabetes aged G, m301G>A >T, i178T>A, 3u1273C>T, and 3u2131C>T) with baseline plasma CRP levels among 1,123 white U.S. participants in the Genetics of Lipid Lowering Drugs and Diet Network (GOLDN) Study and the modulating effect of these SNPs on CRP response to a 3-week fenofibrate treatment among 290 participants with the metabolic syndrome. RESULTS: There were strong associations of m301G>A>T (rs3091244; P = 0.003), i178T>A (rs1417938; P = 0.001), 3u1273C>T (rs1130864; P = 0.001), and 3u2131C>T (rs1205; P < 0.001) with baseline CRP levels. Moreover, among subjects with the metabolic syndrome, fenofibrate induced the greatest reduction in CRP levels for TT subjects of the i178T>A compared with TA and AA subjects (-30 for TT, -19 for TA, and -11% for AA; P = 0.004). Similarly, for the m301G>A>T, major allele carriers displayed maximal reduction of CRP over noncarriers (-20 for GG, 15 for GA and GT, and -0.3% for TA and AA; P = 0.020). CONCLUSIONS: Our results demonstrate that common genetic variants within the CRP gene affect baseline CRP levels and further modulate CRP response in subjects with the metabolic syndrome treated with fenofibrate. This knowledge could contribute to a better prediction of therapeutic success. PMID- 18285552 TI - Estimating physician effects on glycemic control in the treatment of diabetes: methods, effects sizes, and implications for treatment policy. AB - OBJECTIVE: Researchers have only just begun to investigate physician-related effects on medical outcomes. Such research is necessary for developing empirically informed practice guidelines and policy. The primary goal of this study was to investigate whether glucose management in type 2 diabetes varies by randomly assigned physicians over the course of a year in treatment. A second goal of the study was to investigate whether physician-related effects vary across differential patient characteristics. A tertiary goal was to investigate potential patient-level effects on glucose management. RESEARCH DESIGN AND METHODS: Hierarchical linear models were used to investigate A1C among 1,381 patients, nested within 42 randomly assigned primary care physicians at a Veterans Affairs medical center in the southeastern U.S. The primary outcome measure was change in A1C over the course of 1 year in treatment. On average, each study physician had 33 patients with diabetes. RESULTS: Overall, physician related factors were associated with statistically significant but modest variability in A1C change (2%), whereas patient-level factors accounted for the majority of variation in A1C change (98%). Physician effects varied by patient characteristics, mattering more for black patients, patients aged 65 years, and patients whose glucose management improved over the treatment year. CONCLUSIONS: The results of this study indicate that differential physician effects have minimal impact on glycemic control. Results suggest that it is logical to support policies encouraging the development of patient-level behavioral interventions because that is the level that accounts for the majority of variance in glycemic control. PMID- 18285553 TI - Metabolic flexibility in response to glucose is not impaired in people with type 2 diabetes after controlling for glucose disposal rate. AB - OBJECTIVE: Compared with nondiabetic subjects, type 2 diabetic subjects are metabolically inflexible with impaired fasting fat oxidation and impaired carbohydrate oxidation during a hyperinsulinemic clamp. We hypothesized that impaired insulin-stimulated glucose oxidation is a consequence of the lower cellular glucose uptake rate in type 2 diabetes. Therefore, we compared metabolic flexibility to glucose adjusted for glucose disposal rate in nondiabetic versus type 2 diabetic subjects and in the latter group after 1 year of lifestyle intervention (the Look AHEAD [Action For Health in Diabetes] trial). RESEARCH DESIGN AND METHODS: Macronutrient oxidation rates under fasting and hyperinsulinemic conditions (clamp at 80 mU/m(2) per min), body composition (dual energy X-ray absorptiometry), and relevant hormonal/metabolic blood variables were assessed in 59 type 2 diabetic and 42 nondiabetic individuals matched for obesity, sex, and race. Measures were repeated in diabetic participants after weight loss. RESULTS: Metabolic flexibility to glucose (change in respiratory quotient [RQ]) was mainly related to insulin-stimulated glucose disposal rate (R(2) = 0.46, P < 0.0001) with an additional 3% of variance accounted for by plasma free fatty acid concentration at the end of the clamp (P = 0.03). The impaired metabolic flexibility to glucose observed in type 2 diabetic versus nondiabetic subjects (Delta RQ 0.06 +/- 0.01 vs. 0.10 +/- 0.01, respectively, P < 0.0001) was no longer observed after adjusting for glucose disposal rate (P = 0.19). Additionally, the increase in metabolic flexibility to glucose after weight loss was accounted for by the concomitant increase in insulin-stimulated glucose disposal rate. CONCLUSIONS: This study suggests that metabolic inflexibility to glucose in type 2 diabetic subjects is mostly related to defective glucose transport. PMID- 18285554 TI - Asian Indians have enhanced skeletal muscle mitochondrial capacity to produce ATP in association with severe insulin resistance. AB - OBJECTIVE: Type 2 diabetes has become a global epidemic, and Asian Indians have a higher susceptibility to diabetes than Europeans. We investigated whether Indians had any metabolic differences compared with Northern European Americans that may render them more susceptible to diabetes. RESEARCH DESIGN AND METHODS: We studied 13 diabetic Indians, 13 nondiabetic Indians, and 13 nondiabetic Northern European Americans who were matched for age, BMI, and sex. The primary comparisons were insulin sensitivity by hyperinsulinemic-euglycemic clamp and skeletal muscle mitochondrial capacity for oxidative phosphorylation (OXPHOS) by measuring mitochondrial DNA copy number (mtDNA), OXPHOS gene transcripts, citrate synthase activity, and maximal mitochondrial ATP production rate (MAPR). Other factors that may cause insulin resistance were also measured. RESULTS: The glucose infusion rates required to maintain identical glucose levels during the similar insulin infusion rates were substantially lower in diabetic Indians than in the nondiabetic participants (P < 0.001), and they were lower in nondiabetic Indians than in nondiabetic Northern European Americans (P < 0.002). mtDNA (P < 0.02), OXPHOS gene transcripts (P < 0.01), citrate synthase, and MAPR (P < 0.03) were higher in Indians irrespective of their diabetic status. Intramuscular triglyceride, C-reactive protein, interleukin-6, and tumor necrosis factor-alpha concentrations were higher, whereas adiponectin concentrations were lower in diabetic Indians. CONCLUSIONS: Despite being more insulin resistant, diabetic Indians had similar muscle OXPHOS capacity as nondiabetic Indians, demonstrating that diabetes per se does not cause mitochondrial dysfunction. Indians irrespective of their diabetic status had higher OXPHOS capacity than Northern European Americans, although Indians were substantially more insulin resistant, indicating a dissociation between mitochondrial dysfunction and insulin resistance. PMID- 18285555 TI - Urinary Smad1 is a novel marker to predict later onset of mesangial matrix expansion in diabetic nephropathy. AB - OBJECTIVE: We reported that Smad1 is a key transcriptional factor for mesangial matrix expansion in diabetic nephropathy. In this study, we examined whether urinary Smad1 in an early phase of diabetes can predict later development of glomerulosclerosis in diabetic nephropathy and how an angiotensin II type 1 receptor blocker (ARB) can modulate structural changes and urinary markers. RESEARCH DESIGN AND METHODS: Smad1 and albumin in the urine were examined 4 weeks after injection of streptozotocin in 48 rats or 6 weeks of diabetes in db/db mice. Their renal pathology was analyzed after 20 weeks in rats or 12 weeks in mice. Among 48 diabetic rats 7 rats were treated with olmesartan for 20 weeks. RESULTS: Urinary Smad1 of diabetic rats at 4 weeks was nicely correlated with mesangial matrix expansion at 24 weeks (r = 0.70, P < 0.001), while albuminuria showed a weaker association (r = 0.31, P = 0.043). Olmesartan treatment significantly ameliorated glomerulosclerosis and dramatically decreased urinary Smad1 (from 3.9 +/- 2.9 to 0.3 +/- 0.3 ng/mg creatinine, P < 0.05). In db/db mice, urinary Smad1 at 6 weeks was also significantly correlated with mesangial expansion at 18 weeks. In contrast, there was no change in urinary Smad1 in control diabetic rats or mice. CONCLUSIONS: The increase of urinary Smad1 in the early stages of diabetes is correlated with later development of glomerulosclerosis in two rodent models. These data indicate that urinary Smad1 could be a novel predictor for later onset of morphological changes and can be used to monitor the effect of ARBs in diabetic nephropathy. PMID- 18285556 TI - Berberine and its more biologically available derivative, dihydroberberine, inhibit mitochondrial respiratory complex I: a mechanism for the action of berberine to activate AMP-activated protein kinase and improve insulin action. AB - OBJECTIVE: Berberine (BBR) activates AMP-activated protein kinase (AMPK) and improves insulin sensitivity in rodent models of insulin resistance. We investigated the mechanism of activation of AMPK by BBR and explored whether derivatization of BBR could improve its in vivo efficacy. RESEARCH DESIGN AND METHODS: AMPK phosphorylation was examined in L6 myotubes and LKB1(-/-) cells, with or without the Ca(2+)/calmodulin-dependent protein kinase kinase (CAMKK) inhibitor STO-609. Oxygen consumption was measured in L6 myotubes and isolated muscle mitochondria. The effect of a BBR derivative, dihydroberberine (dhBBR), on adiposity and glucose metabolism was examined in rodents fed a high-fat diet. RESULTS; We have made the following novel observations: 1) BBR dose-dependently inhibited respiration in L6 myotubes and muscle mitochondria, through a specific effect on respiratory complex I, similar to that observed with metformin and rosiglitazone; 2) activation of AMPK by BBR did not rely on the activity of either LKB1 or CAMKKbeta, consistent with major regulation at the level of the AMPK phosphatase; and 3) a novel BBR derivative, dhBBR, was identified that displayed improved in vivo efficacy in terms of counteracting increased adiposity, tissue triglyceride accumulation, and insulin resistance in high-fat fed rodents. This effect is likely due to enhanced oral bioavailability. CONCLUSIONS: Complex I of the respiratory chain represents a major target for compounds that improve whole-body insulin sensitivity through increased AMPK activity. The identification of a novel derivative of BBR with improved in vivo efficacy highlights the potential importance of BBR as a novel therapy for the treatment of type 2 diabetes. PMID- 18285557 TI - Plasma free fatty acid storage in subcutaneous and visceral adipose tissue in postabsorptive women. AB - OBJECTIVE: We assessed the direct (VLDL-triglycerides [VLDL-TG] independent) storage of circulating free fatty acids (FFAs) in visceral and subcutaneous fat in postabsorptive women. RESEARCH DESIGN AND METHODS: Twelve women (BMI 29.6 +/- 6.6 kg/m(2)) received an identical, intravenous bolus dose of [1-(14)C]oleate followed by timed subcutaneous fat biopsies (abdominal and femoral) and then omental fat biopsy during tubal ligation surgery. Regional fat masses were assessed by combining dual-energy X-ray absorptiometry and computed tomography scanning. Separately, we assessed the fraction of FFA tracer entering VLDL-TG over the time representing the delay in collecting omental fat. RESULTS: Site specific fat specific activity (SA) (dpm/g lipid) decreased as a function of fat mass in both upper-body subcutaneous (UBSQ) and visceral fat depots. These patterns are consistent with dilution of a relatively fixed amount of FFA tracer within progressively greater amounts of fat. Interestingly, femoral SA did not vary as a function of lower-body subcutaneous (LBSQ) fat mass. [1-(14)C]oleate storage per million LBSQ adipocytes was positively associated with LBSQ fat mass, but no significant relationships were observed in UBSQ or visceral fat depot. The fraction of [1-(14)C]oleate stored in UBSQ, LBSQ, and visceral fat was 6.7 +/- 3.2, 4.9 +/- 3.4, and 1.0 +/- 0.3%, respectively. Only approximately 4% of the tracer traversed VLDL-TG over 9.5 h. CONCLUSIONS: The increase in FFA tracer storage per adipocyte as a function of LBSQ fat mass implies that LBSQ adipocytes, in contrast to UBSQ and omental adipocytes, store more FFA in women with greater adiposity. The direct FFA storage pathway might play a role in favoring lower-body fat accumulation in women. PMID- 18285558 TI - Peroxynitrite mediates retinal neurodegeneration by inhibiting nerve growth factor survival signaling in experimental and human diabetes. AB - OBJECTIVE: Recently we have shown that diabetes-induced retinal neurodegeneration positively correlates with oxidative stress and peroxynitrite. Studies also show that peroxynitrite impairs nerve growth factor (NGF) survival signaling in sensory neurons. However, the causal role of peroxynitrite and the impact of tyrosine nitration on diabetes-induced retinal neurodegeneration and NGF survival signaling have not been elucidated. RESEARCH DESIGN AND METHODS: Expression of NGF and its receptors was examined in retinas from human and streptozotocin induced diabetic rats and retinal ganglion cells (RGCs). Diabetic animals were treated with FeTPPS (15 mg x kg(-1) x day(-1) ip), which catalytically decomposes peroxynitrite to nitrate. After 4 weeks of diabetes, retinal cell death was determined by TUNEL assay. Lipid peroxidation and nitrotyrosine were determined using MDA assay, immunofluorescence, and Slot-Blot analysis. Expression of NGF and its receptors was determined by enzyme-linked immunosorbent assay (ELISA), real-time PCR, immunoprecipitation, and Western blot analyses. RESULTS: Analyses of retinal neuronal death and NGF showed ninefold and twofold increases, respectively, in diabetic retinas compared with controls. Diabetes also induced increases in lipid peroxidation, nitrotyrosine, and the pro-apoptotic p75(NTR) receptor in human and rat retinas. These effects were associated with tyrosine nitration of the pro-survival TrkA receptor, resulting in diminished phosphorylation of TrkA and its downstream target, Akt. Furthermore, peroxynitrite induced neuronal death, TrkA nitration, and activation of p38 mitogen-activated protein kinase (MAPK) in RGCs, even in the presence of exogenous NGF. FeTPPS prevented tyrosine nitration, restored NGF survival signal, and prevented neuronal death in vitro and in vivo. CONCLUSIONS: Together, these data suggest that diabetes-induced peroxynitrite impairs NGF neuronal survival by nitrating TrkA receptor and enhancing p75(NTR) expression. PMID- 18285559 TI - Effectiveness and tolerability of every-other-day rosuvastatin dosing in patients with prior statin intolerance. AB - BACKGROUND: Statins are generally well tolerated, but some patients discontinue therapy secondary to adverse effects. Dosing a statin (rosuvastatin) every other day (EOD) may provide significant lipoprotein changes while avoiding common adverse effects in this statin-intolerant population. OBJECTIVE: To determine the effect and tolerance of EOD rosuvastatin in patients previously intolerant to statin therapy. METHODS: We performed a retrospective analysis of patients treated with EOD rosuvastatin at 2 lipid specialty clinics: the University of Kansas Lipid, Atherosclerosis, and LDL-Apheresis Center and the Hartford Hospital Cholesterol Management Center. Approximately 2600 charts were reviewed to identify patients receiving rosuvastatin EOD who previously had experienced statin intolerance. Fifty-one patients were eligible for the analysis, which evaluated changes in the lipid profile, the number achieving their low-density lipoprotein cholesterol (LDL-C) goals, and the percent tolerating rosuvastatin EOD. Laboratory data were assessed immediately prior to rosuvastatin EOD therapy and at the first follow-up. RESULTS: Myalgias (76.5%) and increased transaminase levels (19.5%) were the most common causes of prior statin intolerance, but 72.5% (37/51) of patients were able to tolerate the EOD therapy (mean dose 5.6 mg) regimen for 4 +/- 2.9 (mean +/- SD) months. Mean LDL-C decreased 34.5% (p < 0.001) in the patients who tolerated the regimen, enabling approximately 50% to achieve their LDL-C goal. All patients who were considered to be intolerant to rosuvastatin EOD therapy (27.5%; 14/51) re-experienced the symptoms of their prior statin intolerance. CONCLUSIONS: Treating patients intolerant to statins with rosuvastatin EOD was tolerated by the majority of patients and reduced LDL-C in our study. This dosing strategy may be useful in patients intolerant to once daily statin dosing, although such an approach has not been documented to reduce cardiovascular events. PMID- 18285560 TI - Nosocomial infections due to methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococcus: relationships with antibiotic use and cost drivers. AB - BACKGROUND: Increased incidence of nosocomial infections due to methicillin resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) has been associated with the use of certain antibiotics and has resulted in increased morbidity, mortality, and costs of care. OBJECTIVE: To describe relationships between vancomycin and linezolid use and incidence of these nosocomial infections over time and to determine factors associated with the increased costs of care (cost drivers) associated with affected patients. METHODS: The association between institution-wide antibiotic use and the rate of nosocomial MRSA and VRE infections was assessed using segmented regression analysis for interrupted time series. The effect that patient characteristics and procedures, as well as certain antibiotic use, had on costs and length of stay of patients with MRSA or VRE nosocomial infection was also assessed and cost drivers for the 2 types of infections were compared. RESULTS: Our analysis included 206 patients who developed MRSA (n = 187) or VRE (n = 19) nosocomial infection. Although small numbers of VRE nosocomial infection may limit generalizations from our results, we found no significant relationship between vancomycin or linezolid use and the rate of either infection. While mean hospital costs were similar, cost drivers varied somewhat between infection types. CONCLUSIONS: The incidence of MRSA or VRE infections does not appear to be related to the use of vancomycin or linezolid. Costs of care are quite high in some affected patients and, while mean total hospital costs are similar, cost drivers appear to differ between the 2 infection types. PMID- 18285561 TI - Oral metoclopramide as an adjunct to analgesics for the outpatient treatment of acute migraine. AB - OBJECTIVE: To compare the combination of oral metoclopramide plus an analgesic with oral triptan monotherapy in the treatment of migraine attacks in outpatients. DATA SOURCES: A search of PubMed (1966-October 2007), EMBASE (1974 October 2007), and International Pharmaceutical Abstracts (1970-October 2007) was conducted using the search terms metoclopramide, migraine, and oral. References of articles identified initially were also reviewed. STUDY SELECTION AND DATA EXTRACTION: Studies evaluating the efficacy and safety of oral metoclopramide compared with oral triptans for migraine treatment were included. Studies of non oral forms of metoclopramide or studies that compared oral metoclopramide with placebo, analgesics, ergotamine, or other migraine therapies were excluded. Studies in which oral metoclopramide was combined with a triptan were also excluded. DATA SYNTHESIS: In the studies identified, oral metoclopramide was combined with aspirin or a similar analgesic. All of the studies assessed headache severity using a 4-point scale (0 = no pain, 1 = mild pain, 2 = moderate pain, 3 = severe pain). Headache relief was defined as a decrease in headache pain from grade 3 or 2 to grade 1 or 0 and was commonly assessed at 2 hours. For all primary and secondary efficacy outcomes-including migraine-associated nausea and vomiting-oral triptans were similar or superior to oral metoclopramide plus an analgesic. The combination was better tolerated, with no reports of serious metoclopramide adverse events such as extrapyramidal symptoms or tardive dyskinesia. CONCLUSIONS: Combinations containing oral metoclopramide plus an analgesic may be an option for patients in whom triptans are contraindicated or who experience intolerable adverse effects, or when cost is an issue. However, patients and physicians should be aware that the combination will likely be less effective than oral triptans in treatment of migraine and its associated symptoms. To determine the role of combination therapy that includes oral metoclopramide in mild-to-moderate migraines, further studies are warranted. PMID- 18285562 TI - Metabolic syndrome and risk of development of atrial fibrillation: the Niigata preventive medicine study. AB - BACKGROUND: The metabolic syndrome consists of a cluster of atherosclerotic risk factors, many of which also have been implicated in the genesis of atrial fibrillation (AF). However, the precise role of the metabolic syndrome in the development of AF is unknown. METHODS AND RESULTS: This prospective, community based, observational cohort study was based on an annual health check-up program in Japan. We studied 28 449 participants without baseline AF. We used 2 different criteria for the metabolic syndrome--the guidelines of the National Cholesterol Education Program Third Adult Treatment Panel (NCEP-ATP III) and those of the American Heart Association/National Heart, Lung, and Blood Institute (AHA/NHLBI)- to study the risk of development of new-onset AF. The metabolic syndrome was present in 3716 subjects (13%) and 4544 subjects (16%) using the NCEP-ATP III and AHA/NHLBI definitions, respectively. During a mean follow-up of 4.5 years, AF developed in 265 subjects (105 women). Among the metabolic syndrome components, obesity (age- and sex-adjusted hazard ratio [HR], 1.64), elevated blood pressure (HR, 1.69), low high-density lipoprotein cholesterol (HR, 1.52), and impaired fasting glucose [corrected] (HR, 1.44 [NCEP-ATP III] and 1.35 [AHA/NHLBI]) showed an increased risk for AF. The association between the metabolic syndrome and AF remained significant in subjects without treated hypertension or diabetes by the NCEP-ATP III definition (HR, 1.78) but not by the AHA/NHLBI definition (HR, 1.28). CONCLUSIONS: The metabolic syndrome was associated with increased risk of AF. The metabolic derangements of the syndrome may be important in the pathogenesis of AF. PMID- 18285563 TI - A quantitative trait locus (LSq-1) on mouse chromosome 7 is linked to the absence of tissue loss after surgical hindlimb ischemia. AB - BACKGROUND: Peripheral arterial disease (PAD) caused by occlusive atherosclerosis of the lower extremity has 2 major clinical manifestations. Critical limb ischemia is characterized by rest pain and/or tissue loss and has a > or = 40% risk of death and major amputation. Intermittent claudication causes pain on walking, has no tissue loss, and has amputation plus mortality rates of 2% to 4% per year. Progression from claudication to limb ischemia is infrequent. Risk factors in most PAD patients overlap. Thus, we hypothesized that genetic variations may be linked to presence or absence of tissue loss in PAD. METHODS AND RESULTS: Hindlimb ischemia (murine model of PAD) was induced in C57BL/6, BALB/c, C57BL/6 x BALB/c (F1), F1 x BALB/c (N2), A/J, and C57BL/6J-Chr7(A/J)/NaJ chromosome substitution strains. Mice were monitored for perfusion recovery and tissue necrosis. Genome-wide scanning with polymorphic markers across the 19 murine autosomes was performed on the N2 mice. Greater tissue loss and poorer perfusion recovery occurred in BALB/c than in the C57BL/6 strain. Analysis of 105 N2 progeny identified a single quantitative trait locus on chromosome 7 that exhibited significant linkage to both tissue necrosis and extent of perfusion recovery. Using the appropriate chromosome substitution strain, we demonstrate that C57BL/6-derived chromosome 7 is required for tissue preservation. CONCLUSIONS: We have identified a quantitative trait locus on murine chromosome 7 (LSq-1) that is associated with the absence of tissue loss in a preclinical model of PAD and may be useful in identifying gene(s) that influence PAD in humans. PMID- 18285564 TI - Cost-effectiveness of providing full drug coverage to increase medication adherence in post-myocardial infarction Medicare beneficiaries. AB - BACKGROUND: Effective therapies for the secondary prevention of coronary heart disease-related events are significantly underused, and attempts to improve adherence have often yielded disappointing results. Elimination of patient out-of pocket costs may be an effective strategy to enhance medication use. We sought to estimate the incremental cost-effectiveness of providing full coverage for aspirin, beta-blockers, angiotensin-converting enzyme inhibitors or angiotensin receptor blockers, and statins (combination pharmacotherapy) to individuals enrolled in the Medicare drug benefit program after acute myocardial infarction. METHODS AND RESULTS: We created a Markov cost-effectiveness model to estimate the incremental cost-effectiveness of providing Medicare beneficiaries with full coverage for combination pharmacotherapy compared with current coverage under the Medicare Part D program. Our analysis was conducted from the societal perspective and considered a lifetime time horizon. In a sensitivity analysis, we repeated our analysis from the perspective of Medicare. In the model, post-myocardial infarction Medicare beneficiaries who received usual prescription drug coverage under the Part D program lived an average of 8.21 quality-adjusted life-years after their initial event, incurring coronary heart disease-related medical costs of $114,000. Those who received prescription drug coverage without deductibles or copayments lived an average of 8.56 quality-adjusted life-years and incurred $111,600 in coronary heart disease-related costs. Compared with current prescription drug coverage, full coverage for post-myocardial infarction secondary prevention therapies would result in greater functional life expectancy (0.35 quality-adjusted life-year) and less resource use ($2500). From the perspective of Medicare, full drug coverage was highly cost-effective ($7182/quality-adjusted life-year) but not cost saving. CONCLUSIONS: Our analysis suggests that providing full coverage for combination therapy to post-myocardial infarction Medicare beneficiaries would save both lives and money from the societal perspective. PMID- 18285565 TI - The Myoblast Autologous Grafting in Ischemic Cardiomyopathy (MAGIC) trial: first randomized placebo-controlled study of myoblast transplantation. AB - BACKGROUND: Phase I clinical studies have demonstrated the feasibility of implanting autologous skeletal myoblasts in postinfarction scars. However, they have failed to determine whether this procedure was functionally effective and arrhythmogenic. METHODS AND RESULTS: This multicenter, randomized, placebo controlled, double-blind study included patients with left ventricular (LV) dysfunction (ejection fraction < or = 35%), myocardial infarction, and indication for coronary surgery. Each patient received either cells grown from a skeletal muscle biopsy or a placebo solution injected in and around the scar. All patients received an implantable cardioverter-defibrillator. The primary efficacy end points were the 6-month changes in global and regional LV function assessed by echocardiography. The safety end points comprised a composite index of major cardiac adverse events and ventricular arrhythmias. Ninety-seven patients received myoblasts (400 or 800 million; n=33 and n=34, respectively) or the placebo (n=30). Myoblast transfer did not improve regional or global LV function beyond that seen in control patients. The absolute change in ejection fraction (median [interquartile range]) between 6 months and baseline was 4.4% (0.2; 7.3), 3.4% (-0.3; 12.4), and 5.2% (-4.4; 11.0) in the placebo, low-dose, and high-dose groups, respectively (P=0.95). However, the high-dose cell group demonstrated a significant decrease in LV volumes compared with the placebo group. Despite a higher number of arrhythmic events in the myoblast-treated patients, the 6-month rates of major cardiac adverse events and of ventricular arrhythmias did not differ significantly between the pooled treatment and placebo groups. CONCLUSIONS: Myoblast injections combined with coronary surgery in patients with depressed LV function failed to improve echocardiographic heart function. The increased number of early postoperative arrhythmic events after myoblast transplantation, as well as the capability of high-dose injections to revert LV remodeling, warrants further investigation. PMID- 18285566 TI - Dietary fish and omega-3 fatty acid consumption and heart rate variability in US adults. AB - BACKGROUND: Fish and omega-3 fatty acid consumption reduce risk of cardiac death, but mechanisms are not well established. Heart rate variability (HRV) predicts cardiac death and reflects specific electrophysiological pathways and influences. We hypothesized that habitual consumption of fish and marine omega-3 fatty acids would be associated with more favorable HRV, elucidating electrophysiological influences and supporting effects on clinical risk. METHODS AND RESULTS: In a population-based cohort of older US adults, we evaluated cross-sectional associations of usual dietary fish and omega-3 consumption during the prior year and ECG-derived (n=4263) and 24-hour Holter monitor-derived (n=1152) HRV. After multivariable adjustment, consumption of tuna or other broiled/baked fish was associated with specific HRV components, including indices suggesting greater vagal predominance and moderated baroreceptor responses (eg, higher root mean square successive differences of normal-to-normal intervals [P=0.001]; higher normalized high-frequency power [P=0.008]; and lower low-frequency/high-frequency ratio [P=0.03]) and less erratic sinoatrial node firing (eg, lower Poincare ratio [P=0.02] and higher short-term fractal scaling exponent [P=0.005]) but not measures of circadian fluctuations (eg, 24-hour standard deviation of normal-to normal intervals). Findings were similar for estimated dietary consumption of marine omega-3 fatty acids. For magnitudes of observed differences in HRV comparing the highest to lowest category of fish intake, differences in relative risk of cardiac death during 10.8 years of follow-up ranged from 1.1% (for difference in standard deviation of normal-to-normal intervals) to 5.9% and 8.4% (for differences in Poincare ratio and short-term fractal scaling exponent) lower risk. CONCLUSIONS: Habitual tuna/other fish and marine omega-3 consumption are associated with specific HRV components in older adults, particularly indices of vagal activity, baroreceptor responses, and sinoatrial node function. Cellular mechanisms and implications for clinical risk deserve further investigation. PMID- 18285567 TI - Microsomal prostaglandin E synthase-1 deletion suppresses oxidative stress and angiotensin II-induced abdominal aortic aneurysm formation. AB - BACKGROUND: Microsomal prostaglandin (PG) E(2) synthase-1 (mPGES-1) catalyzes isomerization of the cyclooxygenase product PGH(2) into PGE(2). Deletion of mPGES 1 modulates experimentally evoked pain and inflammation and retards atherogenesis. The role of mPGES-1 in abdominal aortic aneurysm is unknown. METHODS AND RESULTS: The impact of mPGES-1 deletion on formation of angiotensin II-induced abdominal aortic aneurysm was studied in mice lacking low-density lipoprotein receptor (LDLR(-/-)). Male mice deficient in both mPGES-1 and LDLR (mPGES-1(-/-) LDLR(-/-)) and littermate LDLR(-/-) mice were initiated on a high fat diet at 6 months of age, followed 1 week later by continuous infusion of angiotensin II (1 microg/kg per minute) for an additional 4 weeks. Angiotensin II infusion upregulated aortic expression of cyclooxygenase-2 and mPGES-1, increased aortic macrophage recruitment and vascular nitrotyrosine staining (which reflects local oxidative stress), and augmented urinary excretion of the isoprostane 8,12 iso-iPF(2alpha)-VI (which reflects lipid peroxidation in vivo) and the major metabolite of PGE(2) (PGE-M). Deletion of mPGES-1 decreased both the incidence (87.5% versus 27.3%; P=0.02) and the severity of abdominal aortic aneurysm and depressed the aortic and systemic indices of oxidative stress. Deletion of mPGES 1 also depressed urinary PGE-M, whereas it augmented excretion of PGD(2) and PGI(2) metabolites, reflecting rediversion of the accumulated PGH(2) substrate in the double knockouts. CONCLUSIONS: Deletion of mPGES-1 protects against abdominal aortic aneurysm formation induced by angiotensin II in hyperlipidemic mice, coincident with a reduction in oxidative stress. The potential efficacy of selective inhibition of mPGES-1 in preventing or retarding aneurysm formation warrants further investigation. PMID- 18285568 TI - Cardioprotection by N-acetylglucosamine linkage to cellular proteins. AB - BACKGROUND: The modification of proteins with O-linked beta-N-acetylglucosamine (O-GlcNAc) represents a key posttranslational modification that modulates cellular function. Previous data suggest that O-GlcNAc may act as an intracellular metabolic or stress sensor, linking glucose metabolism to cellular function. Considering this, we hypothesized that augmentation of O-GlcNAc levels represents an endogenously recruitable mechanism of cardioprotection. METHODS AND RESULTS: In mouse hearts subjected to in vivo ischemic preconditioning, O-GlcNAc levels were significantly elevated. Pharmacological augmentation of O-GlcNAc levels in vivo was sufficient to reduce myocardial infarct size. We investigated the influence of O-GlcNAc levels on cardiac injury at the cellular level. Lethal oxidant stress of cardiac myocytes produced a time-dependent loss of cellular O GlcNAc levels. This pathological response was largely reversible by pharmacological augmentation of O-GlcNAc levels and was associated with improved cardiac myocyte survival. The diminution of O-GlcNAc levels occurred synchronously with the loss of mitochondrial membrane potential in isolated cardiac myocytes. Pharmacological enhancement of O-GlcNAc levels attenuated the loss of mitochondrial membrane potential. Proteomic analysis identified voltage dependent anion channel as a potential target of O-GlcNAc modification. Mitochondria isolated from adult mouse hearts with elevated O-GlcNAc levels had more O-GlcNAc-modified voltage-dependent anion channel and were more resistant to calcium-induced swelling than cardiac mitochondria from vehicle mice. CONCLUSIONS: O-GlcNAc signaling represents a unique endogenously recruitable mechanism of cardioprotection that may involve direct modification of mitochondrial proteins critical for survival such as voltage-dependent anion channel. PMID- 18285569 TI - Endothelial cilia are fluid shear sensors that regulate calcium signaling and nitric oxide production through polycystin-1. AB - BACKGROUND: When challenged with extracellular fluid shear stress, vascular endothelial cells are known to release nitric oxide, an important vasodilator. Here, we show that the ability of cultured endothelial cells to sense a low range of fluid shear depends on apical membrane organelles, called cilia, and that cilia are compartments required for proper localization and function of the mechanosensitive polycystin-1 molecule. METHODS AND RESULTS: Cells with the Pkd1(null/null) or Tg737(orpk/orpk) mutation encoded for polycystin-1 or polaris, respectively, are unable to transmit extracellular shear stress into intracellular calcium signaling and biochemical nitric oxide synthesis. Cytosolic calcium and nitric oxide recordings further show that fluid shear sensing is a cilia-specific mechanism because other mechanical or pharmacological stimulation does not abolish calcium and nitric oxide signaling in polycystin-1 and polaris mutant endothelial cells. Polycystin-1 localized in the basal body of Tg737(orpk/orpk) endothelial cells is insufficient for a fluid shear stress response. Furthermore, the optimal shear stress to which the cells respond best does not alter the apical cilia structure but modifies the responsiveness of cells to higher shear stresses through proteolytic modification of polycystin-1. CONCLUSIONS: We demonstrate for the first time that polycystin-1 (required for cilia function) and polaris (required for cilia structure) are crucial mechanosensitive molecules in endothelial cells. We propose that a distinctive communication with the extracellular microenvironment depends on the proper localization and function of polycystin-1 in cilia. PMID- 18285570 TI - CD137 is expressed in human atherosclerosis and promotes development of plaque inflammation in hypercholesterolemic mice. AB - BACKGROUND: Atherosclerosis is a multifactorial disease in which inflammatory processes play an important role. Inflammation underlies lesion evolution at all stages, from establishment to plaque rupture and thrombosis. Costimulatory molecules of the tumor necrosis factor superfamily such as CD40/CD40L and OX40/OX40L have been implicated in atherosclerosis. METHODS AND RESULTS: This study shows that the tumor necrosis factor superfamily members CD137 and CD137 ligand (CD137L), which play a major role in several autoimmune diseases, may constitute a pathogenic pair in atherogenesis. We detected CD137 protein in human atherosclerotic lesions not only on T cells but also on endothelial cells and showed that CD137 in cultured endothelial cells and smooth muscle cells was induced by proinflammatory cytokines implicated in atherosclerosis. Activation of CD137 by CD137L induced adhesion molecule expression on endothelial cells and reduced smooth muscle cell proliferation. In addition, treatment of atherosclerosis-prone apolipoprotein E-deficient mice with a CD137 agonist caused increased inflammation. T-cell infiltration, mainly of CD8(+) cells, and expression of the murine major histocompatibility complex class II molecule I A(b) increased significantly in atherosclerotic lesions, as did the aortic expression of proinflammatory cytokines. CONCLUSIONS: Taken together, these observations suggest that CD137-CD137L interactions in the vasculature may contribute to the progression of atherosclerosis via augmented leukocyte recruitment, increased inflammation, and development of a more disease-prone phenotype. PMID- 18285571 TI - Aortic valvuloplasty in pediatric patients substantially postpones the need for aortic valve surgery: a single-center experience of 188 patients after up to 17.5 years of follow-up. AB - BACKGROUND: Aortic valvuloplasty (AoVP) is an established procedure regarded as a valid alternative for surgical management of congenital aortic valve stenosis. However, its long-term efficacy in preventing or postponing aortic valve surgery remains uncertain for the individual patient. Therefore, the aim of this study was to study the long-term results of AoVP in pediatric patients and its efficacy in preventing or postponing aortic valve surgery. METHODS AND RESULTS: We reviewed up to 17.5 years of follow-up data of all 188 patients who received AoVP at the Deutsches Herzzentrum Munchen. The patients were divided into those < 1 month of age (group < 1 month; n=68) and those > or = 1 month of age (group > or = 1 month; n=120) at the time of AoVP. After the first and second AoVP, moderate and severe aortic regurgitation developed in 29% and 14%, respectively, of the patients in group < 1 month and in 19% and 29%, respectively, of the patients in group > or = 1 month. Survival after 10 years free from aortic valve surgery was 59% (95% confidence interval, 45 to 73) in group < 1 month and 70% (95% confidence interval, 59 to 81) in group > or = 1 month. CONCLUSIONS: This study shows that the long-term results of AoVP of congenital aortic valve stenosis in pediatric patients and its efficacy in preventing or postponing aortic valve surgery are very good. About two thirds of the patients are free from aortic valve surgery 10 years after AoVP. PMID- 18285572 TI - Letter regarding article by Solomon et al, "Cardiac Angiography in REnally Impaired Patients (CARE) study: a randomized double-blind trial of contrast induced nephropathy in patients with chronic kidney disease". PMID- 18285573 TI - Letter regarding article by Solomon et al, "Cardiac Angiography in REnally Impaired Patients (CARE) study: a randomized double-blind trial of contrast induced nephropathy in patients with chronic kidney disease". PMID- 18285574 TI - Pioneers in cardiology: Pedro Brugada, MD, PhD. Interview by Ingrid Torjesen. PMID- 18285575 TI - Public health and the control of high blood pressure at the state level: asleep at the switch or running low on fuel? PMID- 18285576 TI - Bone-marrow-derived matrix metalloproteinase-14: a novel target for plaque stability. PMID- 18285577 TI - Evolving concepts and technologies in mitral valve repair. PMID- 18285578 TI - Contemporary trends in the pharmacological and extracorporeal management of heart failure: a nephrologic perspective. AB - Heart failure and chronic kidney disease share a number of risk factors and pathophysiological pathways. These 2 pathological processes coexist in large numbers of patients. Whereas the presence of chronic kidney disease in patients with heart failure adversely influences their survival, cardiovascular disease is the major cause of mortality in individuals with chronic kidney disease. The management of heart failure by cardiologists has recently expanded from pharmacological treatment to extracorporeal strategies; the interaction between (and concurrent use of) these approaches traditionally has been part of nephrology care and training. The purpose of this review is to explore these management strategies from a nephrologic standpoint and cover the pathophysiology of diuretic resistance, new pharmaceutical strategies to induce natriuresis or aquaresis, and the physiological basis and theoretical advantages of fluid removal by nontraditional peritoneal or hemofiltration approaches. This review also focuses on the technical features, safety, and potential risks of dedicated ultrafiltration devices that do not require dialysis staff or facilities and that are now readily available to nonnephrologists. PMID- 18285579 TI - Images in cardiovascular medicine. Cardiac tuberculoma. PMID- 18285580 TI - The cover. Portrait of Dr William Thomson. PMID- 18285581 TI - A piece of my mind. Sammy. PMID- 18285582 TI - Despite benefit, physicians slow to offer brief advice on harmful alcohol use. PMID- 18285583 TI - Vitamin D deficits may affect heart health. PMID- 18285584 TI - FDA: patients at risk of complications from unretrieved medical device fragments. PMID- 18285585 TI - Treatment regulation in promoting tobacco control. PMID- 18285586 TI - Accounting principles and measuring health care quality. PMID- 18285587 TI - Correction: Inaccurate classification and information reported in a study of statin use and sepsis in patients with chronic kidney disease. PMID- 18285588 TI - Triiodothyronine levels in athyreotic individuals during levothyroxine therapy. AB - CONTEXT: Thyroidal production of triiodothyronine (T3) is absent in athyreotic patients, leading to the suggestion that T3 deficiency may be unavoidable during levothyroxine (LT4) therapy. However, trials evaluating therapy with combined LT4 and T3 have failed to demonstrate any consistent advantage of combination therapy. OBJECTIVE: To determine whether T3 levels in patients treated with LT4 therapy were truly lower than in the same patients with native thyroid function. DESIGN, SETTING, AND PATIENTS: A prospective study conducted in the General Clinical Research Center, Georgetown University Medical Center, Washington, DC, between January 30, 2004, and June 20, 2007, of 50 euthyroid study participants aged 18 to 65 years who were scheduled for total thyroidectomy for goiter, benign nodular disease, suspected thyroid cancer, or known thyroid cancer. Following thyroidectomy, patients were prescribed LT4. Patients with benign thyroid disease and thyroid cancer were treated to achieve a normal and suppressed serum thyroid stimulating hormone (TSH) level, respectively. The LT4 dose was adjusted as necessary postoperatively to achieve the desired TSH goal. MAIN OUTCOME MEASURE: Thyroxine (tetraiodothyronine [T4]), T3, and TSH levels were measured twice preoperatively and twice postoperatively. RESULTS: By the end of the study, there were no significant decreases in T3 concentrations in patients receiving LT4 therapy compared with their prethyroidectomy T3 levels (mean, 127.2 ng/dL; 95% confidence interval [CI], 119.5-134.9 ng/dL vs 129.3 ng/dL; 95% CI, 121.9-136.7 ng/dL; P = .64). However, free T4 concentrations were significantly higher in patients treated with LT(4) therapy (mean, 1.41 ng/dL; 95% CI, 1.33-1.49 ng/dL) compared with their native free T4 levels (1.05 ng/dL; 95% CI, 1.00-1.10 ng/dL; P < .001). Serum TSH values of 4.5 mIU/L or less were achieved in 94% of patients by the end of the study. The T3 concentrations were lower in the subgroup of patients whose therapy had not resulted in a TSH level of 4.5 mIU/L or less (P < .001). CONCLUSION: In our study, normal T3 levels were achieved with traditional LT4 therapy alone in patients who had undergone near-total or total thyroidectomy, which suggests that T3 administration is not necessary to maintain serum T3 values at their endogenous prethyroidectomy levels. PMID- 18285589 TI - Comparison of annual and biannual mass antibiotic administration for elimination of infectious trachoma. AB - CONTEXT: Treatment recommendations assume that repeated mass antibiotic distributions can control, but not eradicate or even locally eliminate, the ocular strains of chlamydia that cause trachoma. Elimination may be an important end point because of concern that infection will return to communities that have lost immunity to chlamydia after antibiotics are discontinued. OBJECTIVE: To determine whether biannual treatment can eliminate ocular chlamydial infection from preschool children and to compare results with the World Health Organization recommended annual treatment. DESIGN, SETTING, AND PARTICIPANTS: A cluster randomized clinical trial of biannual vs annual mass azithromycin administrations to all residents of 16 rural villages in the Gurage Zone, Ethiopia, from March 2003 to April 2005. INTERVENTIONS: At scheduled treatments, all individuals aged 1 year or older were offered a single dose of oral azithromycin either annually or biannually. MAIN OUTCOME MEASURE: Village prevalence of ocular chlamydial infection and presence of elimination at 24 months in preschool children determined by polymerase chain reaction, correcting for baseline prevalence. Antibiotic treatments were performed after sample collections. RESULTS: Overall, 14,897 of 16,403 eligible individuals (90.8%) received their scheduled treatment. In the villages in which residents were treated annually, the prevalence of infection in preschool children was reduced from a mean of 42.6% (range, 14.7% 56.4%) to 6.8% (range, 0.0%-22.0%) at 24 months. In the villages in which residents were treated biannually, infection was reduced from 31.6% pretreatment (range, 6.1%-48.6%) to 0.9% (range, 0.0%-4.8%) at 24 months. Biannual treatment was associated with a lower prevalence at 24 months (P = .03, adjusting for baseline prevalence). At 24 months, no infection could be identified in 6 of 8 of those treated biannually and in 1 of 8 of those treated annually (P = .049, adjusting for baseline prevalence). CONCLUSION: Local elimination of ocular chlamydial infection appears feasible even in the most severely affected areas, although it may require biannual mass antibiotic distributions at a high coverage level. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00221364. PMID- 18285590 TI - Survival from in-hospital cardiac arrest during nights and weekends. AB - CONTEXT: Occurrence of in-hospital cardiac arrest and survival patterns have not been characterized by time of day or day of week. Patient physiology and process of care for in-hospital cardiac arrest may be different at night and on weekends because of hospital factors unrelated to patient, event, or location variables. OBJECTIVE: To determine whether outcomes after in-hospital cardiac arrest differ during nights and weekends compared with days/evenings and weekdays. DESIGN AND SETTING: We examined survival from cardiac arrest in hourly time segments, defining day/evening as 7:00 am to 10:59 pm, night as 11:00 pm to 6:59 am, and weekend as 11:00 pm on Friday to 6:59 am on Monday, in 86,748 adult, consecutive in-hospital cardiac arrest events in the National Registry of Cardiopulmonary Resuscitation obtained from 507 medical/surgical participating hospitals from January 1, 2000, through February 1, 2007. MAIN OUTCOME MEASURES: The primary outcome of survival to discharge and secondary outcomes of survival of the event, 24-hour survival, and favorable neurological outcome were compared using odds ratios and multivariable logistic regression analysis. Point estimates of survival outcomes are reported as percentages with 95% confidence intervals (95% CIs). RESULTS: A total of 58,593 cases of in-hospital cardiac arrest occurred during day/evening hours (including 43,483 on weekdays and 15,110 on weekends), and 28,155 cases occurred during night hours (including 20,365 on weekdays and 7790 on weekends). Rates of survival to discharge (14.7% [95% CI, 14.3%-15.1%] vs 19.8% [95% CI, 19.5%-20.1%], return of spontaneous circulation for longer than 20 minutes (44.7% [95% CI, 44.1%-45.3%] vs 51.1% [95% CI, 50.7%-51.5%]), survival at 24 hours (28.9% [95% CI, 28.4%-29.4%] vs 35.4% [95% CI, 35.0%-35.8%]), and favorable neurological outcomes (11.0% [95% CI, 10.6%-11.4%] vs 15.2% [95% CI, 14.9%-15.5%]) were substantially lower during the night compared with day/evening (all P values < .001). The first documented rhythm at night was more frequently asystole (39.6% [95% CI, 39.0%-40.2%] vs 33.5% [95% CI, 33.2%-33.9%], P < .001) and less frequently ventricular fibrillation (19.8% [95% CI, 19.3%-20.2%] vs 22.9% [95% CI, 22.6%-23.2%], P < .001). Among in-hospital cardiac arrests occurring during day/evening hours, survival was higher on weekdays (20.6% [95% CI, 20.3%-21%]) than on weekends (17.4% [95% CI, 16.8%-18%]; odds ratio, 1.15 [95% CI, 1.09-1.22]), whereas among in-hospital cardiac arrests occurring during night hours, survival to discharge was similar on weekdays (14.6% [95% CI, 14.1% 15.2%]) and on weekends (14.8% [95% CI, 14.1%-15.2%]; odds ratio, 1.02 [95% CI, 0.94-1.11]). CONCLUSION: Survival rates from in-hospital cardiac arrest are lower during nights and weekends, even when adjusted for potentially confounding patient, event, and hospital characteristics. PMID- 18285591 TI - Renal replacement therapy in patients with acute renal failure: a systematic review. AB - CONTEXT: Acute renal failure requiring dialytic support is associated with a high risk of mortality and substantial morbidity. OBJECTIVES: To summarize current evidence guiding provision of dialysis for patients with acute renal failure, to make recommendations for management, and to identify areas in which additional research is needed. DATA SOURCES: Systematic searches of peer-reviewed publications in MEDLINE, EMBASE, and All EBM Reviews through October 2007. STUDY SELECTION: Randomized controlled trials (RCTs) and prospective cohort studies studying dialytic support in adults with acute renal failure that reported the incidence of clinical outcomes such as mortality, length of stay, need for chronic dialysis, or development of hypotension. DATA EXTRACTION: Quality was independently assessed by 2 reviewers using the Jadad score (RCTs) and the Downs and Black checklist (cohort studies). A single reviewer extracted data, which were independently verified by a second reviewer. Results of RCTs were pooled using a random-effects model. DATA SYNTHESIS: From 173 retrieved articles, 30 RCTs and 8 prospective cohort studies were eligible. No conclusions could be drawn about optimal indications for or timing of renal replacement. Available data comparing continuous renal replacement therapy (CRRT) with intermittent hemodialysis demonstrated no clinically relevant difference between modalities, including for all-cause mortality (relative risk [RR], 1.10; 95% confidence interval [CI], 0.99-1.23; I2 = 0%) or for the requirement for chronic dialysis treatment in survivors (RR, 0.91; 95% CI, 0.56-1.49; I2 = 0%). For patients treated with CRRT, limited data suggest that bicarbonate may be preferable to other forms of dialysate alkali and that citrate infusion may be an alternative to systemic anticoagulation in patients at high risk of bleeding. Among patients treated with continuous venovenous hemofiltration (CVVHF), the risk of death was lower at doses of 35 mL/kg per hour (RR of death compared with doses of 20 mL/kg per hour, 0.74; 95% CI, 0.63-0.88). The use of unsubstituted cellulosic membranes should be avoided in intermittent hemodialysis (RR of death compared with biocompatible membranes, 1.23; 95% CI, 1.01-1.50). CONCLUSIONS: Based on current data, intermittent hemodialysis and CRRT appear to lead to similar clinical outcomes for patients with ARF. If CVVHF is used, a dose of 35 mL/kg per hour should be provided. Given the paucity of good-quality evidence in this important area, additional large randomized trials are needed to evaluate clinically important outcomes. PMID- 18285592 TI - Does this patient with diabetes have osteomyelitis of the lower extremity? AB - CONTEXT: Osteomyelitis of the lower extremity is a commonly encountered problem in patients with diabetes and is an important cause of amputation and admission to the hospital. The diagnosis of lower limb osteomyelitis in patients with diabetes remains a challenge. OBJECTIVE: To determine the accuracy of historical features, physical examination, and laboratory and basic radiographic testing. We searched for systematic reviews of magnetic resonance imaging (MRI) in the diagnosis of lower extremity osteomyelitis in patients with diabetes to compare its performance with the reference standard. DATA SOURCES: MEDLINE search of English-language articles published between 1966 and March 2007 related to osteomyelitis in patients with diabetes. Additional articles were identified through a hand search of references from retrieved articles, previous reviews, and polling experts. STUDY SELECTION: Original studies were selected if they (1) described historical features, physical examination, laboratory investigations, or plain radiograph in the diagnosis of lower extremity osteomyelitis in patients with diabetes mellitus, (2) data could be extracted to construct 2 x 2 tables or had reported operating characteristics of the diagnostic measure, and (3) the diagnostic test was compared with a reference standard. Of 279 articles retrieved, 21 form the basis of this review. Data from a single high-quality meta analysis were used to summarize the diagnostic characteristics of MRI in osteomyelitis. DATA EXTRACTION: Two authors independently assigned each study a quality grade using previously published criteria and abstracted operating characteristic data using a standardized instrument. DATA SYNTHESIS: The gold standard for diagnosis is bone biopsy. No studies were identified that addressed the utility of the history in the diagnosis of osteomyelitis. An ulcer area larger than 2 cm2 (positive likelihood ratio [LR], 7.2; 95% confidence interval [CI], 1.1-49; negative LR, 0.48; 95% CI, 0.31-0.76) and a positive "probe-to bone" test result (summary positive LR, 6.4; 95% CI, 3.6-11; negative LR, 0.39; 95% CI, 0.20-0.76) were the best clinical findings. A erythrocyte sedimentation rate of more than 70 mm/h increases the probability of a diagnosis of osteomyelitis (summary LR, 11; 95% CI, 1.6-79). An abnormal plain radiograph doubles the odds of osteomyelitis (summary LR, 2.3; 95% CI, 1.6-3.3). A positive MRI result increases the likelihood of osteomyelitis (summary LR, 3.8; 95% CI, 2.5-5.8). However, a normal MRI result makes osteomyelitis much less likely (summary LR, 0.14; 95% CI, 0.08-0.26). The overall accuracy (ie, the weighted average of the sensitivity and specificity) of the MRI is 89% (95% CI, 83.0% 94.5%). CONCLUSIONS: An ulcer area larger than 2 cm2, a positive probe-to-bone test result, an erythrocyte sedimentation rate of more than 70 mm/h, and an abnormal plain radiograph result are helpful in diagnosing the presence of lower extremity osteomyelitis in patients with diabetes. A negative MRI result makes the diagnosis much less likely when all of these findings are absent. No single historical feature or physical examination reliably excludes osteomyelitis. The diagnostic utility of a combination of findings is unknown. PMID- 18285593 TI - Who is accountable for racial equity in health care? PMID- 18285594 TI - Thyroxine monotherapy after thyroidectomy: coming full circle. PMID- 18285595 TI - Mass antibiotic administration for eradication of ocular Chlamydia trachomatis. PMID- 18285597 TI - JAMA patient page. Osteomyelitis. PMID- 18285600 TI - Should intensity-modulated radiation therapy be the standard of care in the conservatively managed breast cancer patient? PMID- 18285599 TI - Fatigue versus activity-dependent fatigability in patients with central or peripheral motor impairments. AB - In the rehabilitation literature, fatigue is a common symptom of patients with any neurological impairment when defined as a subjective lack of physical and mental energy that interferes with usual activities. Some complaints may, however, arise from fatigability , an objective decline in strength as routine use of muscle groups proceeds. By this refined definition of fatigue, exercise or sustained use reduces the ability of muscles to produce force or power, regardless of whether the task can be sustained. Fatigability may be masked clinically because (1) the degree of weakening is not profound, (2) activity induced weakness rapidly lessens with cessation of exertion, and (3) clinicians rarely test for changes in strength after repetitive movements to objectively entertain the diagnosis. The repetitive movements that induce fatigability during daily activities are an iterative physiological process that depends on changing states induced by activation of spared central and peripheral neurons and axons and compromised muscle. Fatigability may be especially difficult to localize in patients undergoing neurorehabilitation, in part because no finite boundary exists between the central and peripheral components of motor reserve and endurance. At the bedside, however, manual muscle testing before and after repetitive movements could at least put some focus on the presence of fatigability in any patient with motor impairments and related disabilities. Reliable measures of fatigability beyond a careful clinical examination, such as physiological changes monitored by cerebral functional neuroimaging techniques and more standardized central and peripheral electrical and magnetic stimulation paradigms, may help determine the mechanisms of activity-dependent weakening and lead to specific therapies. Testable interventions to increase motor reserve include muscle strengthening and endurance exercises, varying the biomechanical requirements of repetitive muscle contractions, and training-induced neural plasticity or pharmacologic manipulations to enhance synaptic efficacy. PMID- 18285601 TI - Is response rate relevant to the phase II trial design of targeted agents? PMID- 18285602 TI - A multicenter randomized trial of breast intensity-modulated radiation therapy to reduce acute radiation dermatitis. AB - PURPOSE: Dermatitis is a frequent adverse effect of adjuvant breast radiotherapy. It is more likely in full-breasted women and when the radiation is distributed nonhomogeneously in the breast. Breast intensity-modulated radiation therapy (IMRT) is a technique that ensures a more homogeneous dose distribution. PATIENTS AND METHODS: A multicenter, double-blind, randomized clinical trial was performed to test if breast IMRT would reduce the rate of acute skin reaction (notably moist desquamation), decrease pain, and improve quality of life compared with standard radiotherapy using wedges. Patients were assessed each week during and up to 6 weeks after radiotherapy. RESULTS: A total of 358 patients were randomly assigned between July 2003 and March 2005 in two Canadian centers, and 331 were included in the analysis. Breast IMRT significantly improved the dose distribution compared with standard radiation. This translated into a lower proportion of patients experiencing moist desquamation during or up to 6 weeks after their radiation treatment; 31.2% with IMRT compared with 47.8% with standard treatment (P = .002). A multivariate analysis found the use of breast IMRT (P = .003) and smaller breast size (P < .001) were significantly associated with a decreased risk of moist desquamation. The use of IMRT did not correlate with pain and quality of life, but the presence of moist desquamation did significantly correlate with pain (P = .002) and a reduced quality of life (P = .003). CONCLUSION: Breast IMRT significantly reduced the occurrence of moist desquamation compared with a standard wedged technique. Moist desquamation was correlated with increased pain and reduction in the quality of life. PMID- 18285603 TI - Analysis of phase II studies on targeted agents and subsequent phase III trials: what are the predictors for success? AB - PURPOSE: To identify the characteristics of phase II studies that predict for subsequent "positive" phase III trials (those that reached the proposed primary end points of study or those wherein the study drug was superior to the standard regimen investigating targeted agents in advanced tumors. METHODS: We identified all phase III clinical trials of targeted therapies against advanced cancers published from 1985 to 2005. Characteristics of the preceding phase II studies were reviewed to identify predictive factors for success of the subsequent phase III trial. Data were analyzed using the chi(2) test and logistic regression models. RESULTS: Of 351 phase II studies, 167 (47.6%) subsequent phase III trials were positive and 184 (52.4%) negative. Phase II studies from multiple rather than single institutions were more likely to precede a successful trial (60.4% v 39.4%; P < .001). Positive phase II results were more likely to lead to a successful phase III trial (50.8% v 22.5%; P = .003). The percentage of successful trials from pharmaceutical companies was significantly higher compared with academic, cooperative groups, and research institutes (89.5% v 44.2%, 45.2%, and 46.3%, respectively; P = .002). On multivariate analysis, these factors and shorter time interval between publication of phase II results and III study publication were independent predictive factors for a positive phase III trial. CONCLUSION: In phase II studies of targeted agents, multiple- versus single institution participation, positive phase II trial, pharmaceutical company-based trials, and shorter time period between publication of phase II to phase III trial were independent predictive factors of success in a phase III trial. Investigators should be cognizant of these factors in phase II studies before designing phase III trials. PMID- 18285604 TI - Estrogen receptor, progesterone receptor, HER-2, and response to postmastectomy radiotherapy in high-risk breast cancer: the Danish Breast Cancer Cooperative Group. AB - PURPOSE: To examine the importance of estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER-2), and constructed subtypes in a large study randomly assigning patients to receive or not receive postmastectomy radiotherapy (PMRT). PATIENTS AND METHODS: The present analysis included 1,000 of the 3,083 high-risk breast cancer patients randomly assigned to PMRT in the Danish Breast Cancer Cooperative Group (DBCG) protocol 82 trials b and c. Tissue microarray sections were stained for ER, PgR, and HER-2. Median follow-up time for patients alive was 17 years. End points were locoregional recurrence as isolated first event, distant metastases, and overall survival. For statistical analyses four subgroups were constructed from hormonal receptors (Rec). Rec+ was defined as ER+ and/or PgR+. Rec-as both ER-and PgR-. The four subgroups were Rec+/HER-2-, Rec+/HER-2+, Rec-/HER-2-(triple negative), and Rec /HER-2+. RESULTS: A significantly improved overall survival after PMRT was seen only among patients characterized by good prognostic markers such as hormonal receptor-positive and HER-2- patients (including the two Rec+ subtypes). No significant overall survival improvement after PMRT was found among patients with an a priori poor prognosis, the hormonal receptor-negative and HER-2+ patients, and in particular the Rec-/HER-2+ subtype. Furthermore, comparing hazard ratios and 95% CIs, significantly smaller improvements in locoregional recurrence control after PMRT were found for ER-and PgR-tumors compared with the ER+ and PgR+ tumors (P = .003 and .04, respectively), and for the triple-negative (P = .02), and the Rec-/HER-2+ subtypes (P = .003) compared with the Rec+/HER-2 subtype. CONCLUSION: Hormonal receptor status, HER-2, and the constructed subtypes may be predictive of locoregional recurrence and survival after postmastectomy radiotherapy. PMID- 18285605 TI - Lenalidomide for the treatment of B-cell malignancies. AB - Lenalidomide is a novel anticancer agent that has made a major impact in the treatment of patients with B-cell malignancies. A more potent analog of thalidomide, lenalidomide was developed to enhance immunomodulatory properties with improved safety profile. Its antitumor activity seems mediated through modulation of both the cytokine and cellular tumor cell microenvironment. Preclinical as well as clinical observations demonstrate that lenalidomide downregulates production of various critical prosurvival cytokines in the tumor microenvironment while concurrently promoting activation of T- and natural killer (NK) cell-mediated antitumor response. Early clinical investigations noted its efficacy in relapsed and/or refractory multiple myeloma patients. Subsequently, larger randomized studies confirmed the clinical benefit of lenalidomide when added to dexamethasone compared with dexamethasone alone in previously treated myeloma patients resulting in its recent approval by the US Food and Drug Administration. Consequently, the role of lenalidomide in other B-cell malignancies has been investigated, with impressive results in chronic lymphocytic leukemia and non-Hodgkin's lymphoma. This review summarizes the data from various clinical investigations and highlights the impact of lenalidomide in the management of patients with B-cell malignancies. PMID- 18285606 TI - Review of phase II trial designs used in studies of molecular targeted agents: outcomes and predictors of success in phase III. AB - PURPOSE: Because the appropriate design and end points for phase II evaluation of targeted anticancer agents are unclear, we undertook a review of recent reports of phase II trials of targeted agents to determine the types of designs used, the planned end points, the outcomes, and the relationship between trial outcomes and regulatory approval. METHODS: We retrieved reports of single-agent phase II trials in six solid tumors for 19 targeted drugs. For each, we abstracted data regarding planned design and actual results. Response rates were examined for any relationship to eventual success of the agents, as determined by US Food and Drug Administration approval for at least one indication. RESULTS: Eighty-nine trials were identified. Objective response was the primary or coprimary end point in the majority of trials (61 of 89 trials). Fourteen reports were of randomized studies generally evaluating different doses of agents, not as controlled experiments. Enrichment for target expression was uncommon. Objective responses were seen in 38 trials; in 19 trials, response rates were more than 10%, and in eight, they were more than 20%. Agents with high response rates tended to have high nonprogression rates; renal cell carcinoma was the exception to this. Higher overall response rates were predictive of regulatory approval in the tumor types reviewed (P = .005). CONCLUSION: In practice, phase II design for targeted agents is similar to that for cytotoxics. Objective response seems to be a useful end point for screening new targeted agents because, in our review, its observation predicted for eventual success. Improvements in design are recommended, as is more frequent inclusion of biological questions as part of phase II trials. PMID- 18285607 TI - Chemical rescue of deltaF508-CFTR mimics genetic repair in cystic fibrosis bronchial epithelial cells. AB - In a previous study of sodium 4-phenylbutyrate (4-PBA)-responsive proteins in cystic fibrosis (CF) IB3-1 bronchial epithelial cells, we identified 85 differentially expressed high abundance proteins from whole cellular lysate (Singh, O. V., Vij, N., Mogayzel, P. J., Jr., Jozwik, C., Pollard, H. B., and Zeitlin, P. L. (2006) Pharmacoproteomics of 4-phenylbutyrate-treated IB3-1 cystic fibrosis bronchial epithelial cells. J. Proteome Res. 5, 562-571). In the present work we hypothesize that a subset of heat shock proteins that interact with cystic fibrosis transmembrane conductance regulator (CFTR) in common during chemical rescue and genetic repair will identify therapeutic networks for targeted intervention. Immunocomplexes were generated from total cellular lysates, and three subcellular fractions (endoplasmic reticulum (ER), cytosol, and plasma membrane) with anti-CFTR polyclonal antibody from CF (IB3-1), chemically rescued CF (4-PBA-treated IB3-1), and genetically repaired CF (IB3 1/S9 daughter cells repaired by gene transfer with adeno-associated virus-(wild type) CFTR). CFTR-interacting proteins were analyzed on two-dimensional gels and identified by mass spectrometry. A set of 16 proteins known to act in ER associated degradation were regulated in common and functionally connected to the protein processing, protein folding, and inflammatory response. Some of these proteins were modulated exclusively in ER, cytosol, or plasma membrane. A subset of 4-PBA-modulated ER-associated degradation chaperones (GRP94, HSP84, GRP78, GRP75, and GRP58) was observed to associate with the immature B form of CFTR in ER. HSP70 and HSC70 interacted with the C band (mature form) of CFTR at the cell surface. We conclude that chemically rescued CFTR associates with a specific set of HSP70 family proteins that mark therapeutic interactions and can be useful to correct both ion transport and inflammatory phenotypes in CF subjects. PMID- 18285608 TI - Cell stress gives a red light to the mitochondrial cell death pathway. AB - Although the ultimate outcome of prolonged exposure of cells to stress is often death, the early response appears to be the activation of survival pathways that are likely to give the cell an opportunity to repair low-level damage. How these stress-initiated survival pathways influence B cell lymphoma/leukemia 2 (Bcl-2) proteins, the core cell death machinery, has remained unclear; however, two papers now provide insight into stress-mediated survival mechanisms. The liver is unusually resistant to p53-mediated apoptosis. It appears that p53-mediated induction of the gene that encodes insulin-like growth factor-binding protein-1 (IGFBP1) attenuates the cell death response in hepatocytes by preventing the formation of a complex between p53 and the proapoptotic protein BAK. This is especially interesting as IGFBP1 is not a member of the Bcl-2 family, yet it inhibited BAK. In three unrelated cell lines, another regulatory interaction that influences cell survival occurs at the mitochondria. In this case, protein phosphatase 1gamma (PP1gamma) regulated the phosphorylation status of the Bcl 2/Bcl-X(L)-associated death promoter (BAD). The prefoldin family member URI is normally phosphorylated by S6 kinase 1, which liberates PP1gamma from a URI PP1gamma complex. However, the withdrawal of growth factors or nutrients stabilizes this complex, which renders PP1gamma inactive. The net response of this stress stimulus is an increased abundance of phosphorylated BAD, which raises the threshold required to trigger cell death. These two studies have identified new players and mechanisms that integrate stress responses and cell death. PMID- 18285609 TI - The renin rise with aliskiren: it's simply stoichiometry. PMID- 18285610 TI - Lack of association between the type 2 deiodinase A/G polymorphism and hypertensive traits: the Framingham Heart Study. PMID- 18285611 TI - The Atp1a1 gene from inbred Dahl salt sensitive rats does not contain the A1079T missense transversion. AB - The existence of the A1079T transversion in the alpha1 isoform of the Na(+), K(+) ATPase (Atp1a1) gene in Dahl salt-sensitive rat (SS/Jr) strain, discovered by Herrera and Ruiz-Opazo and proposed to underlay hypertension sensitivity, represents one of the most controversial topics in hypertension research. As our research group did not have any previous connection to any party in this dispute nor to hypertension-related research, we were asked (J Hypertens. 2006;24:2312 2313) to definitively adjudge the existence of the A1079T transversion. Hence, different state-of-the art SNP detection technologies that depend on a variety of mechanisms and enzymes to detect the transversion in genomic DNA as well as cDNA derived from different tissues were used. Although it was possible to readily detect other silent polymorphisms between SS and SR strains in the Atp1a1 gene by all methods used, no evidence for the existence of the A1079T transversion in SS/Jr rats was found. PMID- 18285612 TI - Safety of placebo controls in pediatric hypertension trials. AB - Many clinical trials, including those in pediatric populations, use a placebo arm for medical conditions for which there are readily available therapeutic interventions. Several short-term efficacy trials of antihypertensive medications performed in response to Food and Drug Administration-issued written requests have used a placebo arm; whether the use of a placebo arm is safe in children with hypertension is unknown. We sought to define the rates of adverse events in 10 short-term antihypertensive trials to determine whether these trials resulted in increased risk to pediatric patients receiving placebo. We combined patient level data from 10 antihypertensive efficacy trials performed in pediatric patients that were submitted to the Food and Drug Administration from 1998 to 2005. We determined the number and type of all of the adverse events reported during the placebo-controlled portion of the clinical trials and compared these numbers between the patients who received placebo and those who received active drug. Among the 1707 children in the 10 studies, we observed no differences in the rates of adverse events reported between the patients who received placebo and those who received active drug. Only 5 patients suffered a serious adverse event during the trials; none were thought by the investigators to be related to study drug, and only 1 occurred in a patient receiving placebo. Short-term exposure to placebo in pediatric trials of antihypertensive medications appears to be safe. PMID- 18285613 TI - Glucose homeostasis in hypertensive subjects. AB - The objective of this study was to estimate the prevalence of undiagnosed impaired glucose homeostasis in hypertensive subjects in the general population. The most reasonable screening strategy for glucose disorders was also assessed. We carried out an oral glucose tolerance test for 1106 hypertensive subjects aged 45 to 70 years without previously diagnosed diabetes or cardiovascular disease. Blood pressure, waist circumference, body mass index, and plasma lipids were also measured. Type 2 diabetes was found in 66 (6%) of the subjects, impaired glucose tolerance in 220 (20%), and impaired fasting glucose in 167 (15%). If we had carried out an oral glucose tolerance test only for those hypertensive subjects with fasting plasma glucose >or=5.6 mmol/L, we would have missed approximately 40% of the patients with impaired glucose tolerance. The International Diabetes Federation criteria of metabolic syndrome identified 96% of all the cases of type 2 diabetes and 88% of all the cases of impaired glucose tolerance. The prevalence of central obesity was alarming: 90% of the women and 82% of the men had a waist circumference >or=80 cm or >or=94 cm, respectively. Impaired glucose homeostasis and central obesity are common in hypertensive subjects. An oral glucose tolerance test is reasonable to carry out at least for the hypertensive subjects with metabolic syndrome. Weight stabilization is an important goal to treat hypertensive patients. PMID- 18285615 TI - Nitric oxide does not significantly contribute to changes in pulse pressure amplification during light aerobic exercise. AB - NO modulates resting blood pressure and wave reflection. The effect of NO on exercise central hemodynamics is unknown but has important implications relating to cardiovascular risk. The aim of this study was to determine the contribution of NO to pulse pressure (PP) amplification and wave reflection during exercise. Twelve healthy men aged 29+/-1 years (mean+/-SEM) undertook cycle exercise at 60% of their maximal heart rate. Noninvasive measures of central blood pressure, estimated aortic pulse wave velocity, and wave reflection (augmentation index) were obtained by pulse wave analysis during intravenous infusion of saline (control), N(G)-monomethyl-l-arginine (a NO-synthase inhibitor), or noradrenaline (control vasoconstrictor). PP amplification was defined as the ratio of peripheral to central PP. Cardiac output and stroke volume were determined by electric bioimpedance. Both N(G)-monomethyl-l-arginine and noradrenaline caused a significant increase in mean arterial pressure (P<0.01) and augmentation index (P<0.01), as well as reduced ratio of peripheral to central PP (P<0.05) at baseline. Exercise caused a significant increase in the ratio of peripheral to central PP (P<0.001), whereas augmentation index and estimated aortic pulse wave velocity declined (for both P<0.05) during all 3 of the infusion protocols. However, no significant differences were observed in augmentation index, ratio of peripheral to central PP, or estimated aortic pulse wave velocity between infusion procedures (P>0.50) during exercise. Also, heart rate, peripheral vascular resistance, and cardiac output did not differ during exercise between saline, N(G)-monomethyl-l-arginine, or noradrenaline. Although we cannot rule out other vasodilator mechanisms having adjusted for NO blockade, our results indicate that NO does not solely contribute to systemic arterial stiffness or altered blood pressure amplification during light exercise. PMID- 18285616 TI - Statin treatment in hypercholesterolemic pregnant mice reduces cardiovascular risk factors in their offspring. AB - Increasing evidence suggests that hypercholesterolemia during pregnancy initiates pathogenic events in the fetus leading to increased risk of cardiovascular disease in the adult offspring. In this study we examined in mice whether pharmacological intervention using statins in late pregnancy could alleviate the detrimental effects of a high-fat, high-cholesterol (45% fat) maternal diet on the health of the dams and their offspring. Pregnant C57 mice on high-fat, high cholesterol diet were given the 3hydroxy3methylglutaryl-coenzyme A reductase inhibitor pravastatin in the drinking water (5 mg/kg of body weight per day) in the second half of pregnancy and during lactation to lower cholesterol and improve postweaning maternal blood pressure. Weaned offspring were then fed the high-fat, high-cholesterol diet until adulthood (generating dam/offspring dietary groups high-fat, high-cholesterol/high-fat, high-cholesterol and high-fat, high cholesterol plus pravastatin during the second half of pregnancy and lactation/high-fat, high-cholesterol). These groups were compared with offspring from mothers fed standard chow (control), which were then fed control diet to adulthood (control/control). Compared with high-fat, high-cholesterol, high-fat, high-cholesterol plus pravastatin during second half of pregnancy and lactation dams showed significantly reduced total cholesterol concentrations and reduced systolic blood pressure. The high-fat, high-cholesterol plus pravastatin during second half of pregnancy and lactation/high-fat, high-cholesterol offspring were significantly lighter, less hypertensive, and more active compared with the high fat, high-cholesterol/high-fat, high-cholesterol group. Total serum and low density lipoprotein cholesterol concentrations were significantly lower, and high density lipoprotein cholesterol concentrations were raised in high-fat, high cholesterol plus pravastatin during the second half of pregnancy and lactation/high-fat, high-cholesterol offspring, compared with the high-fat, high cholesterol/high-fat, high-cholesterol group. The control/control offspring showed the lowest blood pressure and cholesterol levels. These findings indicate that the cholesterol-lowering effect of statins in pregnant dams consuming a high fat, high-cholesterol diet leads to reduced cardiovascular risk factors in offspring that are sustained into adulthood. PMID- 18285614 TI - Interference with PPARgamma signaling causes cerebral vascular dysfunction, hypertrophy, and remodeling. AB - The transcription factor PPARgamma is expressed in endothelium and vascular muscle where it may exert antiinflammatory and antioxidant effects. We tested the hypothesis that PPARgamma plays a protective role in the vasculature by examining vascular structure and function in heterozygous knockin mice expressing the P465L dominant negative mutation in PPARgamma (L/+). In L/+ aorta, responses to the endothelium-dependent agonist acetylcholine (ACh) were not affected, but there was an increase in contraction to serotonin, PGF(2alpha), and endothelin-1. In cerebral blood vessels both in vitro and in vivo, ACh produced dilation that was markedly impaired in L/+ mice. Superoxide levels were elevated in cerebral arterioles from L/+ mice and responses to ACh were restored to normal with a scavenger of superoxide. Diameter of maximally dilated cerebral arterioles was less, whereas wall thickness and cross-sectional area was greater in L/+ mice, indicating cerebral arterioles underwent hypertrophy and remodeling. Thus, interference with PPARgamma signaling produces endothelial dysfunction via a mechanism involving oxidative stress and causes vascular hypertrophy and inward remodeling. These findings indicate that PPARgamma has vascular effects which are particularly profound in the cerebral circulation and provide genetic evidence that PPARgamma plays a critical role in protecting blood vessels. PMID- 18285619 TI - Relative bioavailability of lisdexamfetamine 70-mg capsules in fasted and fed healthy adult volunteers and in solution: a single-dose, crossover pharmacokinetic study. AB - The relative bioavailability of oral lisdexamfetamine dimesylate, a prodrug of d amphetamine, and active d-amphetamine was assessed in an open-label, single-dose, 3-treatment, 3-period, randomized, crossover study in 18 healthy adult volunteers. Following a fast of at least 10 hours, subjects were administered an intact capsule of 70 mg lisdexamfetamine, a solution containing the capsule contents, or an intact capsule with a high-fat meal. Standard meals started 4 hours following lisdexamfetamine administration. Blood samples were taken predose (0 hours) and 0.5 to 72 hours postdose, and the concentrations of d-amphetamine and lisdexamfetamine were measured. AUC and C(max) for d-amphetamine were similar when lisdexamfetamine 70 mg was administered to healthy adults in the fed or fasted state. The AUC of intact lisdexamfetamine was similar when the latter was taken without food or in solution, but C(max) was lower when lisdexamfetamine was administered with food. The t(max) of d-amphetamine and intact lisdexamfetamine was similar when taken in solution or in the fasted state but was about 1 hour longer when taken with food. Adverse events were typical for amphetamine products. These findings indicate that food does not have a significant effect on d-amphetamine or lisdexamfetamine bioavailability in healthy adults and that lisdexamfetamine was well tolerated. PMID- 18285617 TI - Endogenous melanocortin system activity contributes to the elevated arterial pressure in spontaneously hypertensive rats. AB - Previous studies suggest that activation of the CNS melanocortin system reduces appetite while increasing sympathetic activity and arterial pressure. The present study tested whether endogenous activity of the CNS melanocortin 3/4 receptors (MC3/4-R) contributes to elevated arterial pressure in the spontaneously hypertensive rat (SHR), a model of hypertension with increased sympathetic activity. A cannula was placed in the lateral ventricle of male SHR and Wistar (WKY) rats for chronic intracerebroventricular (ICV) infusions (0.5 muL/h). Mean arterial pressure (MAP) and heart rate (HR) were recorded 24 hour/d using telemetry. After 5-day control period, rats were infused with MC3/4-R antagonist (SHU-9119, 1 nmol/h-ICV) for 12 days, followed by 5-day posttreatment period. MC3/4-R antagonism increased food intake in SHR by 90% and in WKY by 125%, resulting in marked weight gain, insulin resistance, and hyperleptinemia in SHR and WKY. Despite weight gain, MC3/4-R antagonism reduced HR in SHR and WKY ( approximately 40 bpm), while lowering MAP to a greater extent in SHR (-22+/-4 mm Hg) than WKY (-4+/-3 mm Hg). SHU9119 treatment failed to cause further reductions in MAP during chronic adrenergic blockade with propranolol and terazosin. These results suggest that endogenous activity of the CNS melanocortin system contributes to the maintenance of adrenergic tone and elevated arterial pressure in SHR even though mRNA levels for POMC and MC4R in the mediobasal hypothalamus were not increased compared to WKY. These results also support the hypothesis that weight gain does not raise arterial pressure in the absence of a functional MC3/4-R. PMID- 18285620 TI - Clinical microdialysis in skin and soft tissues: an update. AB - Traditionally, plasma or serum drug concentrations have been used for the assessment of bioavailability and bioequivalence. Since in the majority of cases the site of drug action is in the tissue rather than the blood, the use of corresponding free, unbound concentrations in the tissue is a much more meaningful approach. This can become especially important for topical drug administrations, where locally active drug concentrations can significantly exceed free concentrations in plasma. The ability to measure these free concentrations at the site of drug action over time makes microdialysis a very valuable tool for the assessment of bioavailability and bioequivalence. This has been recognized by industry and regulatory authorities, resulting in a recommendation of the microdialysis technique as a tool for bioequivalence determination of topical dermatologic products. The aim of this article is to provide an updated review of the microdialysis technique, its applications in skin and soft tissues, and the resulting impact on clinical drug development. PMID- 18285621 TI - Reconstruction of an object from its symmetry-averaged diffraction pattern. AB - The problem of reconstructing an object from diffraction data that has been incoherently averaged over a discrete group of symmetries is considered. A necessary condition for such data to uniquely specify the object is derived in terms of the object support and the symmetry group. An algorithm is introduced for reconstructing objects from symmetry-averaged data and its use with simulations is demonstrated. The results demonstrate the feasibility of structure determination using a recent proposal for aligning molecules by means of their anisotropic dielectric interaction with an intense light field PMID- 18285622 TI - Higher-dimensional point groups in superspace crystallography. AB - Crystallographic puzzles not covered by the present crystallography, like integral indexing and crystallographic scaling of axial-symmetric biomacromolecules and icosahedral viral capsids and/or integral lattices, can possibly be explained by extending (n,d)-dimensional superspace crystallography to include finite subgroups of the higher-dimensional orthogonal group O(n) and not only those of O(d), as restricted by the physical dimension d. PMID- 18285623 TI - Solving the crystal structures of zeolites using electron diffraction data. I. The use of potential-density histograms. AB - The maximum-entropy and likelihood method for solving zeolite crystal structures from electron diffraction data is modified to use potential-map-density histograms as an additional figure of merit. The experimental histogram is compared to an idealized one (based on known zeolite structures) using Pearson and Spearman correlation coefficients. These supplement the use of log-likelihood estimates as figures of merit to select the optimal solution from a collection of phase sets. The method has been applied with success to seven zeolite and one inorganic crystal structures that have varying associated data quality. The technique works easily even with two-dimensional data sets of less than 50 unique diffraction data and a resolution of less than 2 A. The method is very fast, and the computer time needed on a modest PC was never more than a few minutes. PMID- 18285624 TI - Solving the crystal structures of zeolites using electron diffraction data. II. Density-building functions. AB - A density-building function is used to solve the crystal structures of zeolites from electron diffraction data using both two- and three-dimensional data sets. The observed data are normalized to give unitary structure factors |U(h)|(obs). An origin is defined using one to three reflections and a corresponding maximum entropy map, q(ME)(x), is calculated in which the constraints are the amplitudes and phases of the origin-defining reflections. Eight strong reflections are then given permuted phases and each phase combination is used to compute P(deltaq) = integral(V)deltaq(x)(2)/q(ME)(x)dx, where deltaq(x) is the Fourier transform of |U(h)|(obs)exp(i?phi_(perm)h - |U(h)|(ME)exp(i?phi_(ME)h), phi_(perm)h is the permuted phase for reflection h and phi_(ME)h is the phase angle for reflection h predicted from the Fourier transform of q(ME)(x). The 64 phase sets with minimum values of P(deltaq) are subjected to entropy maximization and, following this procedure, those with the five highest log-likelihood gains are examined. Sometimes auxiliary potential histogram information is also used. The method worked routinely with seven zeolite structures of varying complexity and data quality, but failed with an eighth structure. PMID- 18285625 TI - Crystallography without crystals. I. The common-line method for assembling a three-dimensional diffraction volume from single-particle scattering. AB - It is demonstrated that a common-line method can assemble a three-dimensional oversampled diffracted intensity distribution suitable for high-resolution structure solution from a set of measured two-dimensional diffraction patterns, as proposed in experiments with an X-ray free-electron laser (XFEL) [Neutze et al. (2000). Nature (London), 406, 752-757]. Even for a flat Ewald sphere, it is shown how the ambiguities due to Friedel's law may be overcome. The method breaks down for photon counts below about 10 per detector pixel, almost three orders of magnitude higher than expected for scattering by a 500 kDa protein with an XFEL beam focused to a 0.1 microm diameter spot. Even if 10(3) orientationally similar diffraction patterns could be identified and added to reach the requisite photon count per pixel, the need for about 10(6) orientational classes for high resolution structure determination suggests that about 10(9) diffraction patterns must be recorded. Assuming pulse and readout rates of approximately 100 Hz, such measurements would require approximately 10(7) s, i.e. several months of continuous beam time. PMID- 18285626 TI - Ferroic classifications extended to ferrotoroidic crystals. AB - Aizu's characterization of the 773 species of phase transitions by magnetization, polarization and strain is extended to include characterization by the recently observed toroidal moment. The resulting distinction quadruplet characterization is then used to classify the species into sub-ensembles, extending Schmid's concept of ensembles of species to include classification by toroidal moments. Tables are given of the distinction quadruplet characterization of each species and the species in each ensemble and sub-ensemble. The form of primary and secondary ferroic property tensors invariant under the 122 magnetic point groups have also been tabulated for use in determining the characterization of species and possible domain switching. In both cases, physical property tensors related to the toroidal moment are included. PMID- 18285627 TI - Precise determination of anomalous scattering factors of Ge by using X-ray resonant scattering. AB - Variations of peak position of the rocking curve in the Bragg case are measured from a Ge thin crystal near the K-absorption edge. The variations are caused by a phase change of the real part of the atomic scattering factor. Based on the measurement, the values of the real part are determined with an accuracy of better than 1%. The values are the most reliable ones among those reported values so far as they are directly determined from the normal atomic scattering factors. PMID- 18285628 TI - Minimally resolution biased electron-density maps. AB - Electron-density maps are calculated by Fourier syntheses with coefficients based on structure factors. Diffraction experiments provide intensities up to a limited resolution; as a consequence, the Fourier syntheses always show series termination errors. The worse the resolution, the less accurate is the Fourier representation of the electron density. In general, each atomic peak is shifted from the correct position, shows a deformed (with respect to the true distribution of the electrons in the atomic domain) profile, and is surrounded by a series of negative and positive ripples of gradually decreasing amplitude. An algorithm is described which is able to reduce the resolution bias by relocating the peaks in more correct positions and by modifying the peak profile to better fit the real atomic electron densities. Some experimental tests are performed showing the usefulness of the procedure. PMID- 18285629 TI - A note on Bragg-case Pendellosung and dispersion surface. AB - It is usual to state, since Ewald [Ann. Phys. (1917), 54, 519-597], that Bragg case Pendellosung involves two wavefields belonging to the same branch of the dispersion surface. However, in the Bragg geometry or when crystals are highly absorbing, the dispersion surface can no longer be approximated by the dispersion surface valid for a non-absorbing infinite medium. When considering the real part of the dispersion surface, it is found that the interfering wavefields producing Pendellosung in the Bragg geometry belong in fact to different branches. This paradox is explained by noting that the actual dispersion surface in the semi infinite medium where the wavefields are generated by the incident wave differs from the dispersion surface in an infinite medium. PMID- 18285630 TI - Why phase errors affect the electron function more than amplitude errors. AB - If Fexp(ialpha) are the set of structure factors for a structure f, the amplitudes can be converted to those of an uncorrelated structure g (amplitude swapping) by multiplying each F by the positive number G/F. Correspondingly, the image f is convoluted with k, the Fourier transform of G/F; k has a large peak at the origin, so that f * k approximately f. For swapped phases, the image f is convoluted with l, the Fourier transform of exp(iDeltaalpha), where Deltaalpha, the phase difference between F and G, is a random variable; l does not have a large peak at the origin, so that f * l does not resemble f. The paper provides quantitative descriptions of these arguments. PMID- 18285631 TI - Magnetic group maximal subgroups of index < or =4. AB - The one-, two- and three-dimensional magnetic space groups and the two- and three dimensional magnetic subperiodic groups are considered. The maximal subgroups of index < or =4 of a representative group of each type in the reduced superfamilies of these magnetic groups are tabulated. PMID- 18285634 TI - Ewald Prize award. PMID- 18285636 TI - BDNF increases the phagocytic activity in cultured iris pigment epithelial cells. AB - To investigate the effect of brain derived neurotrophic factor (BDNF) on the phagocytic activity in iris pigment epithelial (IPE) cells, purified porcine photoreceptor outer segments (POS) were applied to cultured IPE cells for three hours. To measure phagocytic activities, bound and total POS were differentially stained using a double immunofluorescence staining method. BDNF increased the binding of POS in IPE cells in a dose-dependent manner. Ingestion of POS, however, was not affected throughout the concentrations used in this study. To investigate the signal transduction pathways of BDNF, a phosphatidylinositol 3 kinase (PI3K) inhibitor, LY294002, and MAPK/ERK kinase (MEK) inhibitor, PD98059, were used for this study. LY294002 had no effect on the binding and ingestion of POS in BDNF-treated IPE cells. On the other hand, PD98059 completely inhibited the increase of POS binding in BDNF-treated cells and also decreased the ingestion of POS. These results indicate that increased POS binding activity by BDNF and the decreased ingestion of POS were mediated through the MAPK pathway. PMID- 18285637 TI - Gene expression and distribution of Swi6 in partial aneuploids of the fission yeast Schizosaccharomyces pombe. AB - Imbalances of gene expression in aneuploids, which contain an abnormal number of chromosomes, cause a variety of growth and developmental defects. Aneuploid cells of the fission yeast Schizosaccharomyces pombe are inviable, or very unstable, during mitotic growth. However, S. pombe haploid cells bearing minichromosomes derived from the chromosome 3 can grow stably as a partial aneuploid. To address biological consequences of aneuploidy, we examined the gene expression profiles of partial aneuploid strains using DNA microarray analysis. The expression of genes in disomic or trisomic cells was found to increase approximately in proportion to their copy number. We also found that some genes in the monosomic regions of partial aneuploid strains increased their expression level despite there being no change in copy number. This change in gene expression can be attributed to increased expression of the genes in the disomic or trisomic regions. However, even in an aneuploid strain that bears a minichromosome containing no protein coding genes, genes located within about 50 kb of the telomere showed similar increases in expression, indicating that these changes are not a secondary effect of the increased gene dosage. Examining the distribution of the heterochromoatin protein Swi6 using DNA microarray analysis, we found that binding of Swi6 within ~50 kb from the telomere occurred less in partial aneuploid strains compared to euploid strains. These results suggest that additional chromosomes in aneuploids could lead to imbalances in gene expression through changes in distribution of heterochromatin as well as in gene dosage. PMID- 18285638 TI - Body distribution of inhaled fluorescent magnetic nanoparticles in the mice. AB - Reducing the particle size of materials is an efficient and reliable tool for improving the bioavailability of a gene or drug delivery system. In fact, nanotechnology helps in overcoming the limitations of size and can change the outlook of the world regarding science. However, a potential harmful effect of nanomaterial on workers manufacturing nanoparticles is expected in the workplace and the lack of information regarding body distribution of inhaled nanoparticles may pose serious problem. In this study, we addressed this question by studying the body distribution of inhaled nanoparticles in mice using approximately 50-nm fluorescent magnetic nanoparticles (FMNPs) as a model of nanoparticles through nose-only exposure chamber system developed by our group. Scanning mobility particle sizer (SMPS) analysis revealed that the mice were exposed to FMNPs with a total particle number of 4.89 x 10(5) +/- 2.37 x 10(4)/cm(3) (low concentration) and 9.34 x 10(5) +/- 5.11 x 10(4)/cm(3) (high concentration) for 4 wk (4 h/d, 5 d/wk). The body distribution of FMNPs was examined by magnetic resonance imaging (MRI) and Confocal Laser Scanning Microscope (CLSM) analysis. FMNPs were distributed in various organs, including the liver, testis, spleen, lung and brain. T2-weighted spin-echo MR images showed that FMNPs could penetrate the blood-brain-barrier (BBB). Application of nanotechnologies should not produce adverse effects on human health and the environment. To predict and prevent the potential toxicity of nanomaterials, therefore, extensive studies should be performed under different routes of exposure with different sizes and shapes of nanomaterials. PMID- 18285639 TI - Assessment of DNA damage in Japanese nurses handling antineoplastic drugs by the comet assay. AB - To clarify genotoxic effects of occupational exposure to antineoplastic drugs in Japan, we examined DNA damage, assessed by the comet assay, in 121 female nurses and 46 female clerks working at three hospitals in the northeast of Japan. The comet assay is considered to be a sensitive and rapid method for DNA strand break detection in individual cells, and tail length and tail moment are used as the comet parameters. Concerning the basal characteristics, the 46 control subjects had higher rates of smoking and coffee-drinking habits and lower hemoglobin than the 121 nurses (p<0.05). The log-transformed tail length in the nurses was significantly longer than that in the control subjects after adjusting for possible covariates such as age and smoking habit (p<0.05). Also, the log transformed tail length was significantly longer, in the 57 nurses who had handled antineoplastic drugs in the last six months, than that in the 46 control subjects (p<0.05); but, no significant difference in tail length or tail moment was seen between the two nurse groups with and without experience of handling hazardous drugs (p>0.05). These results suggest that Japanese nurses who have worked at hospitals using antineoplastic drugs may have a potential risk of DNA damage. To minimize this risk in Japan, use of biological safety cabinet and appropriate protective equipment, in addition to staff education and training, should be implemented in the healthcare environment. PMID- 18285640 TI - Perception in relation to a potential influenza pandemic among healthcare workers in Japan: implications for preparedness. AB - Due to the potential for an influenza pandemic, preparedness for infection control in healthcare settings is essential from the standpoint of occupational health for healthcare workers. We conducted questionnaire surveys among Japanese hospitals to assess preparedness at the individual and institutional levels and their inter-relationship. Questionnaires were administered at 7 tertiary hospitals in Japan during the spring of 2006. We analyzed 7,378 individual responses of the 10,746 questionnaires administered and all seven institutional responses by hospital infection control committees. Healthcare workers assigned low importance to personal protective equipment and showed mixed attitudes (anxious but accepting) to the potential risk. Institutional gaps existed in preparedness across hospitals and most hospitals lacked the specificity to cope with a pandemic. A higher level of institutional preparedness, as determined by expertise as well as general and specific countermeasures, was an important predictor of individual recognition of preventive measures, perception of institutional measures, and attitude toward coping with risk. A higher level of institutional preparedness stood out to be an important predictor of individual preparedness. Considering the risk of a future influenza pandemic, hospitals should improve preparedness at all levels. PMID- 18285641 TI - Relationships between self-rating of recovery from work and morning salivary cortisol. AB - To date, the understanding of how recovery from work relates to cortisol output is poor. Considering this, the present study set out to investigate the associations between self-ratings of 15 items of rest and recovery and salivary cortisol sampled every second hour across two working days. Data came from 12 female and 13 male white-collar workers and were analyzed by linear regression analyses and repeated measures ANOVA. Poor rest and recovery was associated with high levels of morning cortisol, with the strongest relationships emerging for "rested in the morning", "rested after a weekend", "feel energetic during the working day", "tired during the working day", "sufficient sleep" and "worry about something". Moreover, significant interaction effects emerged between sex and "rested after a weekend" and "worry about something". To conclude, the findings show that self-ratings of rest and recovery are related to cortisol, particularly to morning cortisol, and that self-ratings provide important information on physiological recovery in terms of cortisol output. PMID- 18285642 TI - Association of physical job demands, smoking and alcohol abuse with subsequent premature mortality: a 9-year follow-up population-based study. AB - This study assessed the relationships of physical job demands (PJD), smoking, and alcohol abuse, with premature mortality before age 70 (PM-70) among the working or inactive population. The sample included 4,268 subjects aged 15 or more randomly selected in north-eastern France. They completed a mailed questionnaire (birth date, sex, weight, height, job, PJD, smoking habit, alcohol abuse (Deta questionnaire)) in 1996 and were followed for mortality until 2004 (9 yr). PJD score was defined by the cumulative number of the following high job demands at work: hammer, vibrating platform, pneumatic tools, other vibrating hand tools, screwdriver, handling objects, awkward posture, tasks at heights, machine tools, pace, working on a production line, standing about and walking. The data were analyzed using the Poisson regression model. Those with PM-70 were 126 (3.81 per 1,000 person-years). The leading causes of death were cancers (46.4% in men, 57.1% in women), cardiovascular diseases (20.2% and 11.9%), suicide (9.5% and 7.1%), respiratory diseases (6.0% and 4.8%), and digestive diseases (2.4% and 4.8%). PJD3, smoker, and alcohol abuse had adjusted risk ratios of 1.71 (95% CI 1.02-2.88), 1.76 (1.08-2.88), and 2.07 (1.31-3.26) respectively for all-cause mortality. Manual workers had a risk ratio of 1.84 (1.00-3.37) compared to the higher socio-economic classes. The men had a two-fold higher mortality rate than the women; this difference became non-significant when controlling for job, PJD, smoker and alcohol abuse. For cancer mortality the factors PJD3, smoker, and alcohol abuse had adjusted risk ratios of 2.00 (1.00-3.99), 2.34 (1.19-4.63), and 2.22 (1.17-4.20), respectively. Health promotion efforts should be directed at structural measures of task redesign and they should also concern lifestyle. PMID- 18285643 TI - Short-term effects of ambient gaseous pollutants and particulate matter on daily mortality in Shanghai, China. AB - Identification of the specific pollutants contributing most to the health hazard of the air pollution mixture may have important implications for environmental and social policies. In the current study, we conducted a time-series analysis to examine the specific effects of major air pollutants [particulate matter less than 10 microns in diameter (PM(10)), sulfur dioxide (SO(2)), and nitrogen dioxides (NO(2))] on daily mortality in Shanghai, China, using both single pollutant and multiple-pollutant models. In the single-pollutant models, PM(10), SO(2), and NO(2) were found to be associated with mortality from both all non accidental causes and from cardiopulmonary diseases. Unlike some prior studies in North America, we found a significant effect of gaseous pollutants (SO(2) and NO(2)) on daily mortality even after adjustment for PM(10) in the multiple pollutant models. Our findings, combined with previous Chinese studies showing a consistent, significant effect of gaseous pollutants on mortality, suggest that the role of outdoor exposure to SO(2) and NO(2) should be investigated further in China. PMID- 18285644 TI - Evaluation of occupational exposure to antiblastic drugs in an Italian hospital oncological department. AB - The determination of the current antiblastic drug contamination levels in an Italian hospital oncology ward was carried out. Statistical evaluation of data aiming to identify potential exposure causes was performed. Cyclophosphamide (CP), ifosfamide (IF) and 5-fluorouracil (5-FU) were determined by wipe tests, extracted with diatomaceous earths and quantified by GC/MSMS or HPLC/UV. Data were analysed with respect to the potential contamination levels of sampled surfaces, and various amounts of handled analyte. chi(2) tests and Spearman correlation coefficients were calculated. Median concentration levels of 0.086, 0.071, 2.363 microg/dm(2) were obtained, (CP, IF, 5-FU, respectively). 3.8 and 13.5% of investigated surfaces showed CP and IF concentrations higher than 1 microg/dm(2) (up to 26.96 microg/dm(2)) and 13.4% of samples contained 5-FU concentrations in the range 20-208.9 microg/dm(2). Analytes' concentration levels were dependent on sampling sites, with significant correlations showing a progressive contamination decrement going from workbenches, floor, hood planes and other surfaces. A diffuse contamination (traces of all the three analytes) was found on all investigated surfaces, even when analytes had not been used during the sampling days. A significant correlation (rho s=0.303, p=0.001) between the measured analyte concentration and the analyte handled amount was found only in the case of IF. The risk management strategy should be improved, as suggested by the measured and widespread levels of contamination. Since contamination also depends on other factors attributable to working modalities and cleaning procedures, the obtained results suggest that performance of specific training courses as well as scheduling environmental monitoring plans to achieve an actual decrement of the observed contamination levels should be implemented. PMID- 18285645 TI - Postural support by a standing aid alleviating subjective discomfort among cooks in a forward-bent posture during food preparation. AB - In this study, we evaluated the effects on subjective discomfort among cooks during food preparation through use of a standing aid that we developed to alleviate the workload on the low back in the forward-bent posture. Twelve female cooks who worked in a kitchen in a nursing home were asked to prepare foods in 2 working postures: (a) supported by the standing aid (Aid) and (b) without the aid (No aid). They were instructed to evaluate discomfort in 13-body regions during food preparation and the degree of fatigue at the day's end and to enter their ratings after the end of the workday. Since a significant correlation was observed between body height and the improvement effect of discomfort through use of the standing aid, cooks were divided into two groups according to the height, and ratings were analyzed in each group. Among the tall cooks, subjective discomfort in the low back and the front and back of thighs was significantly less with the Aid posture than with the No aid posture. However, in short cooks these values tended to increase in the Aid posture compared with the No aid posture. The results suggest that the standing aid was effective in alleviating tall cooks' workload on the low back in the forward-bent posture. PMID- 18285646 TI - Psychological job characteristics and alexithymic traits in Korean white-collar workers. PMID- 18285647 TI - Reliability and validity of the Japanese version of the Stanford Presenteeism Scale in female employees at 2 Japanese enterprises. PMID- 18285648 TI - Direct medical costs and working days lost due to non-fatal occupational injuries in Denizli, Turkey. PMID- 18285649 TI - Simultaneous observation of zero-value of FSBP% and Raynaud's phenomenon during cold provocation in vibration syndrome. PMID- 18285650 TI - Association of job-related stress factors with psychological and somatic symptoms among Japanese hospital nurses: effect of departmental environment in acute care hospitals. AB - The present study examined degrees of job-related stress factors as well as mental and physical symptoms among Japanese hospital nurses in various departments, and clarified associations of departments and job-related stress factors with those symptoms. A self-administered anonymous questionnaire was distributed to 1,882 full-time nurses at four acute care hospitals in Japan. The survey included demographic factors, and the Brief Job Stress Questionnaire. Among 1,599 nurses who completed all items relevant to the present study, we analyzed data from 1,551 female nurses. The results show that working in operating rooms was associated with fatigue, that working in intensive care units (ICU) was associated with anxiety, and that working in surgery and internal medicine was associated with anxiety and depression independently of demographic factors and job-related stress factors. The physical and mental health of nurses might affect their time off, quality of nursing care and patient satisfaction in acute care hospitals. Therefore, job-related stress factors should be minimized, to improve the physical and mental health of nurses, considering unique departmental demands. PMID- 18285651 TI - Actual conditions of the mixing of antineoplastic drugs for injection in hospitals in Osaka Prefecture, Japan. AB - We conducted a questionnaire survey in order to grasp the actual conditions under which antineoplastic drugs are mixed for injection in hospitals. Questionnaires were sent to all 155 hospitals with 100 or more beds for general patients in Osaka Prefecture, Japan. The response rate was 69.0%. Mixing of antineoplastic drugs was done in 81.3% of the hospitals. The questionnaire was answered by doctors in 17.2% of the hospitals with antineoplastic drugs, nurses in 11.5%, and pharmacists in 70.1%. Mixing of antineoplastic drugs was done by doctors in 58.6% of the hospitals, nurses in 44.8%, and pharmacists in 63.2% (multiple answers). Occupational exposure to antineoplastic drugs was recognized in 97.7% of the hospitals. The mean frequency of the mixing operation was 8.8 d per month per worker. The mean number of antineoplastic drugs handled was 7.4 types. Guidelines for the safe handling of antineoplastic drugs were used in 52.8% of the hospitals and a biological safety cabinet was available in 57.4%. Gloves, mask, gown and goggles were used in 82.7, 69.0, 62.1 and 36.8% of the hospitals, respectively, but no personal protective equipment was used in 10.1%. The safety precautions of the hospitals in which the number of beds was small tended to be fewer than those of the hospitals in which the number of beds was large. Used vials and ampoules were disposed of as clinical, infectious or exclusive antineoplastic drug waste by 74.7% of the hospitals. Safety measures for handling the excrement of patients treated with antineoplastic drugs were performed in 8.0% of the hospitals. In 43.7% of the hospitals, the responders had experienced accidents during antineoplastic drug preparation, such as drugs adhering to hands or eyes, drug leakage, accidental injection and cutting by ampoules. Because of the adverse effects of antineoplastic drugs, all hospitals in which the healthcare workers handle them should promote safety precautions. PMID- 18285652 TI - Occupational stress and mental health among correctional officers: a cross sectional study. AB - A harsh psychosocial environment in the workplace might cause adverse health events, but the association has not been well demonstrated in the penitentiary environment. This cross-sectional study was designed to explore the association between workplace psychosocial risks and the mental health of correctional officers in a Spanish penitentiary center. A self-administered questionnaire was distributed to correctional officers. A total of 164 responded anonymously (response rate 43%). The SF36 survey was used to measure mental health and ISTAS21 (Spanish version of Copenhagen Psychosocial Questionnaire) was used to measure exposure to workplace psychosocial conditions. Low scores were obtained for mental health, high scores were obtained for psychological demands, low self esteem, and low control and influence and moderate scores for low social support, double exposure, and insecurity at work. A linear regression analysis was constructed to study the influence of workplace psychosocial conditions (independent variables) on mental health (dependent variable). The effect was adjusted for sex, age, seniority, and occupational group. Psychological demands (highest impact), low control and influence, and double exposure had significant inverse associations with mental health. The association between low social support, low self-esteem, and insecurity at work with mental health was insignificant. Psychosocial work conditions are a potential target for mental health promotion programs at work. PMID- 18285653 TI - Basic and clinical aspects of non-neuronal acetylcholine: expression of non neuronal acetylcholine in urothelium and its clinical significance. AB - Recently, several reports demonstrate that non-neuronal acetylcholine (ACh) release may contribute to various pathophysiological conditions. In this review, we presented our experiments designed to evaluate the non-neuronal cholinergic system in human bladder. After insertion of the microdialysis probe, human bladder strips were suspended in an organ bath filled with Krebs-Henseleit solution, and Ringer solution was perfused into the probe. ACh release was measured by microdialysis and HPLC. The contribution of urothelium and the effects of age and stretch of bladder strips on non-neuronal ACh release were evaluated. Choline acetyltransferase (ChAT) immunohistochemical staining of bladder was also performed. Immunohistochemistry showed marked ChAT-positive staining in the urothelium. There was tetrodotoxin-insensitive non-neuronal ACh release and this was significantly higher in strips with urothelium than in strips without urothelium. The non-neuronal ACh release was increased with age. Stretch of bladder strips caused increases in non-neuronal ACh release. The stretch-induced release of non-neuronal ACh was increased with age. Our data demonstrate that there is a non-neuronal cholinergic system in human bladder and that urothelium contributes to non-neuronal ACh release. There was significant age-related and stretch-induced increase in non-neuronal ACh release. It is suggested that the non-neuronal cholinergic system may contribute to the physiology and pathophysiology of human bladder. We also discussed the clinical significance of the non-neuronal cholinergic system in human bladder. PMID- 18285654 TI - Basic and clinical aspects of non-neuronal acetylcholine: expression of an independent, non-neuronal cholinergic system in lymphocytes and its clinical significance in immunotherapy. AB - Lymphocytes possess all the components required to constitute an independent, non neuronal cholinergic system. These include acetylcholine (ACh); choline acetyltransferase (ChAT), its synthesizing enzyme; and both muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively). ACh modifies T and B cell function via both mAChR- and nAChR-mediated pathways. Stimulation of lymphocytes with the T cell activator phytohemagglutinin, protein kinase C activator phorbol ester, or cell surface molecules enhances the synthesis and release of ACh and up-regulates ChAT and/or M(5) mAChR gene expression. Furthermore, animal models of immune disorders exhibit abnormal lymphocytic cholinergic activity. The cholesterol-lowering drug simvastatin attenuates the lymphocytic cholinergic activity of T cells by inhibiting LFA-1 signaling in a manner independent of its cholesterol-lowering activity. This suggests that simvastatin exerts its immunosuppressive effects in part by modifying lymphocytic cholinergic activity. Nicotine, an active ingredient of tobacco, ameliorates ulcerative colitis but exacerbates Crohn's disease. Expression of mRNAs encoding the nAChR alpha7 and alpha5 subunits are significantly diminished in peripheral mononuclear leukocytes from smokers, as compared with those from nonsmokers. In addition, long-term exposure of lymphocytes to nicotine reduces intracellular Ca(2+) signaling via alpha7 nAChR-mediated pathways. In fact, studies of humoral antibody production in M(1)/M(5) mAChR-deficient and alpha7 nAChR-deficient animals revealed the role of lymphocytic cholinergic activity in the regulation of immune function. These results provide clues to understanding the mechanisms underlying immune system regulation and could serve as the basis for the development of new immunomodulatory drugs. PMID- 18285655 TI - Basic and clinical aspects of non-neuronal acetylcholine: expression of non neuronal acetylcholine in lung cancer provides a new target for cancer therapy. AB - Lung cancer is the leading cause of cancer death worldwide and new treatment strategies are clearly needed. The recent discovery that lung and other cancers synthesize and secrete acetylcholine (ACh) which acts as an autocrine growth factor suggests that this cholinergic autocrine loop may present new therapeutic targets. In normal bronchial epithelium, small airway epithelium and pulmonary neuroendocrine cells synthesize Ach; and in squamous cell lung carcinoma, adenocarcinoma, and small cell lung carcinoma, the respective lung cancers that derive from those cell types similarly synthesize ACh. ACh secreted by those cancers stimulates growth of the tumors by binding to nicotinic and muscarinic receptors expressed on lung cancers. Thus antagonists to nicotinic and muscarinic receptors can inhibit lung cancer growth. The muscarinic receptor (mAChR) subtype utilized for cell proliferation is the M(3) subtype and consistent with this M(3) mAChR antagonists inhibit growth of SCLC and squamous cell carcinomas. This is significant as M(3) mAChR antagonists have low toxicity and are in wide clinical use. As multiple other cancer types besides lung carcinomas express both M(3) mAChR and acetylcholine, other cancer types besides lung carcinoma may respond to M(3) mAChR antagonists. PMID- 18285656 TI - Basic and clinical aspects of non-neuronal acetylcholine: biological and clinical significance of non-canonical ligands of epithelial nicotinic acetylcholine receptors. AB - Mucocutaneous keratinocytes and bronchial epithelial cells express nicotinic acetylcholine receptors (nAChRs). Emerging evidence indicates that nAChRs can be stimulated also by the tobacco-derived nitrosamines 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) that can induce tumors in laboratory animals. Nitrosamines may disturb the delicate balance between cell proliferation, growth arrest, and apoptosis. A novel paradigm of cell regulation via nAChR has been discovered in studies of SLURP (secreted mammalian Ly 6/urokinase plasminogen activator receptor-related protein)-1 and -2. Experimental results suggest that SLURP-1 and -2 regulate keratinocyte proliferation, apoptosis, and differentiation. Most importantly, SLURPs and professional nicotinic antagonists can abolish, in part, the abilities of NNK and NNN to cause tumorigenic transformation of immortalized keratinocytes. Learning the pharmacology of the nitrosamine vs. SLURP action on epithelial cells may help develop an effective anti-cancer treatment and prevention programs wherein hazardous effects of tobacco products are anticipated, or even abolished, by a pharmacologic ligand of the specific nicotinic receptor acting as an antidote. PMID- 18285657 TI - Basic and clinical aspects of non-neuronal acetylcholine: overview of non neuronal cholinergic systems and their biological significance. AB - Acetylcholine (ACh) is a phylogenetically ancient molecule involved in cell-to cell signaling in almost all life-forms on earth. Cholinergic components, including ACh, choline acetyltransferase, acetylcholinesterase, and muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively) have been identified in numerous non-neuronal cells and tissues, including keratinocytes, cancer cells, immune cells, urinary bladder, airway epithelial cells, vascular endothelial cells, and reproductive organs, among many others. Stimulation of the mAChRs and nAChRs elicits cell-specific functional and biochemical effects. These findings support the notion that non-neuronal cholinergic systems are expressed in certain cells and tissues and are involved in the regulation of their function and that cholinergic dysfunction is related to the pathophysiology of certain diseases. They also provide clues for development of drugs with novel mechanisms of action. PMID- 18285658 TI - Current topics in DNA double-strand break repair. AB - DNA double strand break (DSB) is one of the most critical types of damage which is induced by ionizing radiation. In this review, we summarize current progress in investigations on the function of DSB repair-related proteins. We focused on recent findings in the analysis of the function of proteins such as 53BP1, histone H2AX, Mus81-Eme1, Fanc complex, and UBC13, which are found to be related to homologous recombination repair or to non-homologous end joining. In addition to the function of these proteins in DSB repair, the biological function of nuclear foci formation following DSB induction is discussed. PMID- 18285659 TI - Hyperoxia exposure induced hormesis decreases mitochondrial superoxide radical levels via Ins/IGF-1 signaling pathway in a long-lived age-1 mutant of Caenorhabditis elegans. AB - The hormetic effect, which extends the lifespan by various stressors, has been confirmed in Caenorhabditis elegans (C. elegans). We have previously reported that oxidative stress resistance in a long-lived mutant age-1 is associated with the hormesis. In the age-1 allele, which activates an insulin/insulin-like growth factor-1 (Ins/IGF-1) signaling pathway, the superoxide dismutase (SOD) and catalase activities increased during normal aging. We now demonstrate changes in the mitochondrial superoxide radical (*O(2)(-)) levels of the hormetic conditioned age-related strains. The *O(2)(-) levels in age-1 strain significantly decreased after intermittent hyperoxia exposure. On the other hand, this phenomenon was not observed in a daf-16 null mutant. This hormesis-dependent reduction of the *O(2)(-) levels was observed even if the mitochondrial Mn-SOD was experimentally reduced. Therefore, it is indicated that the hormesis is mediated by events that suppress the mitochondrial *O(2)(-) production. Moreover, some SOD gene expressions in the hormetic conditioned age-1 mutant were induced over steady state mRNA levels. These data suggest that oxidative stress-inducible hormesis is associated with a reduction of the mitochondrial *O(2)(-) production by activation of the antioxidant system via the Ins/IGF-1 signaling pathway. PMID- 18285660 TI - Fullerenol C60(OH)24 effects on antioxidative enzymes activity in irradiated human erythroleukemia cell line. AB - Radiotherapy-induced toxicity is a major dose-limiting factor in anti-cancer treatment. Ionizing radiation leads to the formation of reactive oxygen and nitrogen species (ROS/RNS) that are associated with radiation-induced cell death. Investigations of biological effects of fullerenol have provided evidence for its ROS/RNS scavenger properties in vitro and radioprotective efficiency in vivo. Therefore we were interested to evaluate its radioprotective properties in vitro in the human erythroleukemia cell line. Pre-treatment of irradiated cells by fullerenol exerted statistically significant effects on cell numbers and the response of antioxidative enzymes to X-ray irradiation-induced oxidative stress in cells. Our study provides evidence that the pre-treatment with fullerenol enhanced the enzymatic activity of superoxide dismutase and glutathione peroxidase in irradiated K562 cells. PMID- 18285661 TI - Inverse dose-rate-effects on the expressions of extra-cellular matrix-related genes in low-dose-rate gamma-ray irradiated murine cells. AB - Based on the results of previous microarray analyses of murine NIH3T3/PG13Luc cells irradiated with continuous low-dose-rate (LDR) gamma-ray or end-high-dose rate-irradiations (end-HDR) at the end of the LDR-irradiation period, the inverse dose-rate-effects on gene expression levels were observed. To compare differences of the effects between LDR-irradiation and HDR-irradiation, HDR-irradiations at 2 different times, one (ini-HDR) at the same time at the start of LDR-irradiation and the other (end-HDR), were performed. The up-regulated genes were classified into two types, in which one was up-regulated in LDR-, ini-HDR-, and end-HDR irradiation such as Cdkn1a and Ccng1, which were reported as p53-dependent genes, and the other was up-regulated in LDR- and ini-HDR irradiations such as pro collagen TypeIa2/Col1a2, TenascinC/Tnc, and Fibulin5/Fbln5, which were reported as extra-cellular matrix-related (ECM) genes. The time dependent gene expression patterns in LDR-irradiation were also classified into two types, in which one was an early response such as in Cdkn1a and Ccng1 and the other was a delayed response such as the ECM genes which have no linearity to total dose. The protein expression pattern of Cdkn1a increased dose dependently in LDR- and end-HDR irradiations, but those of p53Ser15/18 and MDM2 in LDR-irradiations were different from end-HDR-irradiations. Furthermore, the gene expression levels of the ECM genes in embryonic fibroblasts from p53-deficient mice were not increased by LDR- and end-HDR-irradiation, so the delayed expressions of the ECM genes seem to be regulated by p53. Consequently, the inverse dose-rate-effects on the expression levels of the ECM genes in LDR- and end-HDR-irradiations may be explained from different time responses by p53 status. PMID- 18285662 TI - Regeneration of extrahepatic bile duct--possibility to clinical application by recognition of the regenerative process. AB - The biliary epithelium is continuously exposed to highly cytotoxic bile acids and pathogens and thus is at persistent risk for injury. The monolayer mucosal epithelium protects the body from these dangers and once injured. The bile duct repair process essentially involves reconstruction of the bile duct with migrating cells, but there are many questions about the process. It is reported that implantation of a bioabsorbable polymer tube as a bypass graft into the extrahepatic bile duct resulted in bile duct regeneration in the graft site after the artificial duct had been degraded and absorbed. We briefly describe our findings on extrahepatic biliary tissue regeneration with the possibility for clinical applications in mind. The creation of this artificial bile duct may be able to promote the development of the treatment for biliary diseases. PMID- 18285663 TI - Properties of acetylcholine-induced hyperpolarization in smooth muscle cells of the mouse mesenteric artery. AB - The properties of smooth muscle cell hyperpolarization produced by acetylcholine (ACh) were investigated in mesenteric arteries isolated from mice. The resting membrane potential of the smooth muscle cells was about -60 mV. When ACh (10 microM) was applied for 1 min, the membrane hyperpolarized with a peak amplitude of about 5 mV which was reached in about 1 min, following which the potential slowly reverted to the resting level over about 7 min following withdrawal of ACh from the superfusate (recovery component). Exposure of the artery to 0.5 mM Ba(2+), an inhibitor of inward rectifier K-channels, depolarized the membrane by about 13 mV, increased the amplitude of the ACh-induced hyperpolarization to about 10 mV, and facilitated the visualization of the recovery component. Indomethacine (10 microM), an inhibitor of cyclooxygenase, inhibited the recovery component and as a consequence reduced the duration of the hyperpolarization. The ACh-induced response was not markedly altered by either N(omega)-nitro-L-arginine (10 microM), an inhibitor of nitric oxide (NO) production, or catalase (130 U/ml), a super oxide scavenger. Exogenously applied hydrogen peroxide (H(2)O(2), 300 microM) hyperpolarized the membrane by about 5 mV, which was abolished by catalase. These results suggest that in the mouse mesenteric artery, the ACh induced hyperpolarization has two components, both an indomethacin-sensitive and an indomethacin-insensitive component. The former component may be produced by prostanoids, while the latter may be produced by factors other than NO or H(2)O(2). The results also suggested that the inward rectifier K-channels may be important for producing the resting membrane potential, but they may not be the main contributor to the ACh-induced hyperpolarization of smooth muscle cell membranes in the mouse mesenteric artery. PMID- 18285664 TI - Mechanical and electrical responses modulated by excitation of inhibitory nerves during stimulation with high-potassium solutions in circular smooth muscle of the rabbit rectum. AB - The properties of the mechanical responses produced by solutions containing high concentrations of potassium ion (high-K solution, [K(+)](o) = 9-27 mM) were investigated in circular smooth muscle preparations isolated from the rabbit rectum. Isometric recording of mechanical responses of the muscle revealed spontaneous contractions, which successively decreased and finally disappeared in most preparations. Stimulation of the smooth muscle with high-K solutions elicited an increase in both amplitude and frequency of twitch contractions (sustained component), with about a 2 min delay in the beginning (initial inhibition), and a transient large contraction shortly after the cessation of stimulation (after contraction). Transmural nerve stimulation (TNS) with electrical pulses for 1 min at 1 Hz frequency produced a sustained inhibition, but a transient contraction followed after termination of TNS. In the presence of tetrodotoxin (TTX), the TNS-induced responses were abolished, while a high-K solution elicited increased twitch contractions with a short delay and abolished the after contraction. Suramin produced effects similar to TTX on the responses produced by high-K solutions or TNS, but this was not the case for atropine, guanethidine or N(omega)-nitro-L-arginine (L-NA). Recording membrane potentials with microelectrodes revealed that TNS evoked an inhibitory junction potential (i.j.p.) which was non-adrenergic, non-cholinergic and non-nitrergic in nature. High-K solutions elicited a tri-phasic change in the membrane potential; an initial hyperpolarization, followed by a sustained depolarization and finally a transient depolarization on cessation of high-K stimulation. TTX or suramin inhibited the i.j.p.s and altered the tri-phasic change in the membrane potential produced by a high-K solution to a mono-phasic depolarization. No significant modulation of electrical responses of the membrane induced by TNS or high-K solution was elicited by atropine, guanethidine or L-NA. The results indicated that the circular smooth muscle of the rabbit rectum is innervated by inhibitory nerves, and that stimulation with high-K solutions caused inhibitory neuronal modulation of both electrical and mechanical responses of the smooth muscle, in a suramin-sensitive way. PMID- 18285665 TI - Altered biochemical parameters in the saliva of patients with breast cancer. AB - Saliva plays an important role in the protection of oral cavity and alterations in either salivary flow rate or protein composition may have dramatic effects on oral health. Prevention and management of oral complications of cancer and cancer therapy will improve oral function and quality of life, and reduce morbidity and the cost of care. The aim of this study was to investigate the saliva of patients with breast cancer biochemically and cytologically and compare with healthy controls. Accordingly, lipid peroxidation (LPO), total protein, salivary flow rate, and pH levels were measured in the saliva samples obtained from 20 breast cancer patients and 11 healthy individuals. Tissue factor (TF) is a major regulator of normal hemostasis and thrombosis, and TF activity of saliva samples was evaluated. Under the conditions used, patients with breast cancer present a significant reduction in total protein, pH and LPO levels. Salivary TF activity was higher in breast cancer patients than that in control subjects, but the degree of increase was not statistically significant. In addition, the analysis of saliva samples by SDS polyacrylamide gel electrophoresis showed the retarded mobility of the 66-kDa proteins and the increased proteins of about 36 kDa in the patient group. Some patients with breast cancer had increased number of leucocytes. Importantly, dysplastic cells and yeast cells were detected only in saliva samples of cancer patients. Decreased salivary LPO may be considered as a risk factor for breast cancer. PMID- 18285666 TI - Interlaboratory comparison of quantitative RT-PCR based detection for minimal residual disease in leukemias: a standardization approach in Japan. AB - Real-time quantitative polymerase chain reaction (RQ-PCR) has been accepted as integral part of the management of patients with hematologic malignancies. Whereas standardization efforts of RQ-PCR, initiated by Europe Against Cancer (EAC) group, have been gradually widespread in the world, Japanese laboratories use their individual protocol for RQ-PCR analysis. Therefore, we assessed the variability of quantitative results obtained from 4 different laboratories in Japan, including 3 companies and Tohoku University Hospital, using identical peripheral blood or bone marrow samples of patients in chronic myeloid leukemia (CML; n = 11) and acute myeloid leukemia (AML; n = 2). RQ-PCR was designed to quantify the copy numbers of disease-specific fusion chimeras; BCR-ABL (CML) and AML1-ETO (AML). In 5 out of 13 samples, the quantitative results from 4 laboratories varied more than 10 times (up to 712 times). Thus, we next sought to determine factors affecting the variability of RQ-PCR results across laboratories, by sending back RNA and cDNA samples from each company to Tohoku University, and they were further proceed to yield quantitative data. The main difference between companies and Tohoku University was probably due to the difference of blood separation method (Blood lysis or Ficoll-Hypaque). On the other hand, the variability among 4 laboratories was the most noticeable in the PCR step, mainly attributable to the difference of primer/probe sequence among laboratories. In conclusion, our analyses indicate the importance to limit both preanalytical (sample processing) and analytical (RQ-PCR) interlaboratory variability for RQ-PCR protocol, and the need of further efforts on standardization program in Japan. PMID- 18285667 TI - Development of a new method for estimating visceral fat area with multi-frequency bioelectrical impedance. AB - Excessive visceral fat area (VFA) is a major risk factor in such conditions as cardiovascular disease. In assessing VFA, computed tomography (CT) is adopted as the gold standard; however, this method is cost intensive and involves radiation exposure. In contrast, the bioelectrical impedance (BI) method for estimating body composition is simple and noninvasive and thus its potential application in VFA assessment is being studied. To overcome the difference in obtained impedance due to measurement conditions, we developed a more precise estimation method by selecting the optimum body posture, electrode arrangement, and frequency. The subjects were 73 healthy volunteers, 37 men and 36 women, who underwent CT scans to assess VFA and who were measured for anthropometry parameters, subcutaneous fat layer thickness, abdominal tissue area, and impedance. Impedance was measured by the tetrapolar impedance method using multi-frequency BI. Multiple regression analysis was conducted to estimate VFA. The results revealed a strong correlation between VFA observed by CT and VFA estimated by impedance (r = 0.920). The regression equation accurately classified VFA > or = 100 cm(2) in 13 out of 14 men and 1 of 1 woman. Moreover, it classified VFA > or = 100 cm(2) or < 100 cm(2) in 3 out of 4 men and 1 of 1 woman misclassified by waist circumference (W) which was adopted as a simple index to evaluate VFA. Therefore, using this simple and convenient method for estimating VFA, we obtained an accurate assessment of VFA using the BI method. PMID- 18285668 TI - Effectiveness of oseltamivir treatment among children with influenza A or B virus infections during four successive winters in Niigata City, Japan. AB - Oseltamivir has been used for treatment of influenza A and B infections, but recent reports documented that it was less active against the latter. We compared the effectiveness of oseltamivir in children between laboratory confirmed influenza A and B over 4 influenza seasons from 2001 to 2005 in a pediatric clinic in Japan. Among 1,848 patients screened, 299 influenza A and 209 influenza B patients were administered oseltamivir (treated groups), and 28 influenza A and 66 influenza B patients were assigned as non-treated groups. The duration of fever, defined as period when patients had the maximum temperature higher than 37.5 degrees C in three-time measurements in a day after the clinic visit, was evaluated among the four groups. In uni-variate analysis, the duration of fever was shorter for treated group than non-treated for influenza A (1.8 +/- 0.9 days vs 2.6 +/- 1.3 days, p < 0.01), but it was not significant for influenza B (2.4 +/- 1.3 days vs 2.8 +/- 1.2 days, p = 0.9). The fever duration was longer in treated influenza B than A patients (p < 0.01). Multi-variate analysis indicated younger age (< 6 years old) and higher body temperature at the clinic visit prolonged the duration of fever. Adjusted average duration of fever indicated that oseltamivir was effective for both types, but more effective on influenza A, and the benefit increased for younger children. Our data provide evidence that oseltamivir is beneficial for influenza infections, but the effectiveness is differed by type and age. PMID- 18285669 TI - Antimicrobial effect of fluoroquinolones for the eradication of nontypeable Haemophilus influenzae isolates within biofilms. AB - Biofilms can be defined as communities of microorganisms attached to a surface. Those bacterial biofilms cause serious problems, such as antibiotic resistance and medical device-related infections. Nontypeable Haemophilus influenzae (NTHi) is an important pathogen in respiratory infections, as it forms biofilms both in vitro and in vivo such as human middle ear. Recent reports indicate that otitis media, paranasal sinusitis and lower respiratory tract infections caused by Haemophilus influenzae have become more difficult to treat with oral antibiotic therapy. However, there has been no attention given to antibiotic eradication of NTHi biofilm. To investigate the antimicrobial effect of various antibiotics against NTHi biofilm formation, we conducted the following comparative study using both beta-lactamase-negative ampicillin (AMP)-susceptible (BLNAS) and AMP resistant (BLNAR) NTHi strains. In a microtiter biofilm assay, both levofloxacin and gatifloxacin, of the fluoroquinolone antibiotic group, significantly inhibited biofilm formation by BLNAS and BLNAR NTHi in a dose-dependent fashion compared to ampicillin of the penicillin antibiotic group, cefotaxime of the cephalosporin antibiotic group, and both erythromycin and clarithromycin of the macrolide antibiotic group. Furthermore, in flow cell chamber studies, confocal laser scanning microscopy counted survival bacteria in mature biofilm had been treated with gatifloxacin, ampicillin, cefotaxime and erythromycin. Only gatifloxacin completely killed the BLNAR NTHi isolates within biofilms without regard to the thickness of biofilm formation. The results of this study suggest that fluoroquinolones potentially have a role in therapy against diseases caused by both BLNAS and BLNAR NTHi isolates within biofilms. PMID- 18285670 TI - Recurrence and persistence of fever in children who developed amantadine resistant influenza viruses after treatment. AB - In recent years, a dramatic increase of amantadine-resistant influenza A has occurred globally, but limited data have been available on the clinical course of patients developed amantadine-resistant viruses. We compared fever reduction between patients who developed resistance or remained sensitive in a pediatric clinic in Niigata, Japan, from 2000 to 2006. A total of 2,802 clinical samples were collected from patients who visited the pediatric outpatient clinic with influenza like illness during the seven influenza epidemic seasons. Patients were divided into 4 groups and analyzed for the fever reduction after amantadine treatment: emerged amantadine-resistant (n = 15); amantadine-sensitive (n = 35); patients administered no antiviral drugs (n = 42); and oseltamivir-treated patients (n = 320), which served as references. All 4 groups showed alleviation of fever up to day 3. The amantadine-resistant group had a significant recurrence of fever on day 4 and/or 5, and as a consequence, the course of illness was prolonged. Considering the pattern of fever, recurrent and persistent patterns were found significantly at higher rates in children with emerged resistant virus compared to other groups, and the age tended to be younger in amantadine resistant compared to amantadine-sensitive group (3.9 +/- 3.0 vs 6.7 +/- 4.1 years old, n.s.). Therefore, we concluded that younger children were prone to develop amantadine-resistance after treatment and showed a significant recurrence of fever on day 4 and/or 5, and the course of illness was consequently prolonged. PMID- 18285671 TI - Increased expression of mucin 5AC mRNA and decreased expression of epidermal growth-factor receptor mRNA in gallstone patients. AB - Mucin, a major component of mucus, plays an important role in gallstone formation. The molecular mechanisms of mucin overproduction, however, still remain unknown. Several mucin genes (MUC) have been implicated in various diseases and gel-forming mucin genes (MUC2, MUC5AC, MUC5B, and MUC6) were recognized to be the important components of digestive mucus. Furthermore epidermal growth factor receptor (EGFR) might regulate the function of MUC5AC. The aim of this study was to investigate the expression of MUC5AC mRNA and the possible role of EGFR in the function of MUC5AC. Total twelve patients underwent elective laparoscopic cholecystectomy due to gallstone disease were enrolled (age: 64.6 +/- 15.5 years). The control group included two patients who underwent cholecystectomy without stone. The expression levels of MUC5AC and EGFR mRNAs were analyzed in gallbladder tissues using real-time reverse transcription polymerase chain reaction assay. The expression levels of MUC5AC mRNA were increased in gallstone patients compared with those in control subjects, ranging from 2.5 to 1558 folds (mean 512.8 +/- 493.6 folds, p = 0.004). In contrast, the expression levels of EGFR mRNA were decreased in all patients (mean 0.378 +/- 0.322 fold, p = 0.002), and negative correlation was found between MUC5AC and EGFR (p = 0.02). There was no significant difference according to age, body mass index, and stone composition in the expression levels of MUC5AC and EGFR mRNAs. In conclusion, MUC5AC is over-expressed in gallstone disease, despite the decrease in the expression of EGFR mRNA. MUC5AC may be related to mucus hypersecretion. PMID- 18285672 TI - Normal blood flow velocities of basal cerebral arteries decrease with advancing age: a transcranial Doppler sonography study. AB - Transcranial Doppler sonography (TCD) is a non-invasive diagnostic tool enabling evaluation of blood flow characteristics of basal intracerebral vessels via thin calvarian regions. Several factors may affect the normal values of cerebral hemodynamic parameters, and standard reference values for each laboratory are needed for precise interpretation of the results. The aims of this study were to determine normal values of flow velocities of basal cerebral arteries of our TCD laboratory, and to study the influence of age and gender on normal values. We studied 63 healthy volunteers (30 male and 33 females; age range, 5 - 69 years old) with TCD with a 2-MHz transcranial probe. The subjects were divided into 7 age groups: 5-10 years, 11-20 years, 21-30 years, 31-40 years, 41-50 years, 51-60 years and > 60 years. Mean velocity (V mean), peak systolic velocity (PSV), and end-diastolic velocities (EDV) were determined in middle, anterior and posterior cerebral arteries. No significant gender difference was found. However, there was a decrease in blood flow velocities in all vessels with advancing age, which was significant when subjects older than 40 years and < or = 40 years old were compared. V mean, PSV and EDV values were highest in the age group of 5 - 10 years old and lowest in volunteers older than the age of 60 (p < 0.05). As a conclusion, flow velocities in basal cerebral arteries range widely and are significantly age-related. Age matching of TCD data is a requirement for clinically relevant conclusions. PMID- 18285673 TI - Extracorporeal shock wave therapy ameliorates hindlimb ischemia in rabbits. AB - We have recently demonstrated that the low-energy extracorporeal cardiac shock wave (SW) therapy improves myocardial perfusion and cardiac function in a porcine model of chronic myocardial ischemia and also ameliorates myocardial ischemia in patients with severe coronary artery disease. The present study was designed to examine whether our SW therapy also is effective to ameliorate hindlimb ischemia in rabbits. Hindlimb ischemia was made by surgical excision of the entire unilateral rabbit femoral artery. One week after the operation, we performed the SW (n = 9) or sham-therapy (n = 9) to the ischemic region 3 times a week for 3 weeks. Three weeks after the SW therapy, the development of collateral arteries, the flow ratio of the ischemic/non-ischemic common iliac arteries, the blood pressure ratio of the ischemic/non-ischemic hindlimb, and the capillary density in the ischemic muscles were all significantly increased in the SW group compared with the control group, indicating that the SW therapy induced therapeutic angiogenesis. Importantly, no adverse effect, such as muscle damage, hemorrhage, or thrombosis, was noted with the therapy. Finally, we examined the role of endothelial nitric oxide synthesis (eNOS) and vascular endothelial growth factor (VEGF) in the mechanisms of SW-induced angiogenesis on day 28. The expression levels of eNOS and VEGF proteins in ischemic hindlimb muscles tended to be increased in the SW group compared with the control group. These results suggest that our low-energy SW therapy also is effective and safe for the treatment of peripheral artery disease. PMID- 18285674 TI - Allogeneic peripheral blood stem cell transplantation from related donors mismatched at 2 HLA loci in the host-versus-graft direction. AB - In allogeneic stem cell transplantation, immune reactions can occur in 2 directions. The recipient's lymphocytes can recognize the donor's cells as "foreign" and attempt to kill them, which results in the host-versus-graft (HVG) reaction that is commonly termed graft rejection. The other direction is the graft-versus-host (GVH) reaction. When the recipient is homozygous at a mismatched human leukocyte antigen (HLA) locus, HLA disparity is present only in the former direction and not in the latter direction. If transplants harvested from such an HVG-mismatched donor can be used to achieve stable engraftment with minimal toxicity, then these donors can potentially be a useful alternative donor source. Here, we report 2 patients (1 with acute myeloblastic leukemia and another with lymphoblastic lymphoma) who were transplanted with peripheral blood stem cells (PBSCs) obtained from related donors mismatched at 2 HLA loci in the HVG direction but completely matched in the GVH direction. Our conditioning regimen, consisting of busulfan, cyclophosphamide, low-dose total body irradiation (TBI) (4 Gy), and fludarabine, achieved successful engraftment with an acceptable level of regimen-related toxicity. Our experience suggests that PBSC transplantation with an HVG-mismatched related donor and an appropriate conditioning regimen may be a therapeutic option for patients in whom early transplantation is desirable. PMID- 18285675 TI - Zarit burden inventory and activities of daily living in the behavioral variant of frontotemporal dementia. AB - BACKGROUND: Activities of daily living (ADL) and caregiver burden are known to have a major impact on the decision to institutionalize patients with Alzheimer's disease (AD), yet little research has been done on these aspects in patients with frontotemporal dementia (FTD). AIM: To compare ADL and caregiver burden in FTD and in early-onset AD. METHODS: We compared 26 FTD and 28 AD patients with respect to the Neuropsychiatric Inventory (NPI), Mini Mental State Examination, Mattis Dementia Rating Scale (MDRS), Disability Assessment for Dementia (DAD) and Zarit Burden Inventory (ZBI). RESULTS: Demographic variables for FTD and AD were similar. FTD patients obtained a significantly higher NPI behavioral score than AD patients (median, 39.5 vs. 11; p < 0.0001). However, the two groups did not differ in their total DAD score. No correlations were observed between DAD and cognitive status (MDRS) or between DAD and behavioral impairment (NPI). The ZBI was higher in FTD than in AD patients (median, 40 vs. 18.5; p = 0.0004) and was correlated with the NPI in both groups. CONCLUSION: Functional disability was similar in FTD and AD patients. Nevertheless, the caregiver burden was higher in FTD than in AD, a result that has important implications for caregiver help. PMID- 18285676 TI - Role of biologics and other therapies in stricturing Crohn's disease: what have we learnt so far? AB - BACKGROUND: Therapy of strictures, one of the most common complications of Crohn's disease (CD), remains a challenging task in gastroenterology. While infliximab is widely recognized as being very effective in active CD, it has been reported to cause strictures in some patients. As a consequence, essentially by inference, many clinicians have chosen not to use it in the presence of strictures. AIMS: To find evidence in the available data that infliximab does not cause strictures and that there is no rational basis to avoid its a priori use when a stricture is already present. In addition, to review what is currently known on the general management of strictures in CD. METHODS: Discussion of the data that led to the hypothesis of a causal association between infliximab and strictures. Review of the mechanisms and the risk factors for stricture development in CD; of the different types of CD-related strictures; of the available means to distinguish them, and of the literature related to the efficacy and safety of infliximab as well as other biologics and other therapies in different stricturing scenarios. RESULTS AND CONCLUSIONS: Although larger controlled studies are due in the near future, current evidence indicates that infliximab does not cause strictures in CD. The drug appears safe and effective in the presence of an inflammatory stenosis while being predictably ineffective, but not harmful, in the presence of fibrosis. Different stricturing scenarios in CD must be clearly distinguished for proper management of this complication. PMID- 18285677 TI - Usefulness of the short IQCODE for predicting postoperative delirium in elderly patients undergoing hip and knee replacement surgery. AB - BACKGROUND/OBJECTIVE: The prevalence of postoperative delirium in elderly patients is >30%. The objective of this prospective study was to determine the usefulness of the short form of the Informant Questionnaire on COgnitive Decline in the Elderly (short IQCODE) to predict the occurrence of postoperative delirium after elective hip and knee arthroplasty in the elderly. METHODS: Consecutive patients, 60 years and older, who were admitted for elective hip or knee arthroplasty were included. The preoperative cognitive status was determined using the Mini-Mental State Examination (MMSE) and the short IQCODE. Postoperative delirium was diagnosed using the Confusion Assessment Method. Logistic regression was used to analyze the links between the preoperative test scores and the outcome of postoperative delirium. RESULTS: One hundred and one patients completed the study (mean age 73.6 +/- 6.6 years). The mean +/- SD MMSE score was 26 +/- 3, and the mean short IQCODE score was 50.7 +/- 6.2. Postoperative delirium developed in 15 patients (14.8%). A short IQCODE score >50 was significantly associated with postoperative delirium (OR 12.7, 95% CI 1.4 115.5; p = 0.02). CONCLUSIONS: The short IQCODE appears to be a useful tool to predict the risk of postoperative delirium in elderly patients undergoing elective surgery. Detecting this complication could be of great interest to improve the postoperative survey of elderly patients. PMID- 18285678 TI - Treatment with combined streptozotocin and liposomal doxorubicin in metastatic endocrine pancreatic tumors. AB - Treatment with combined streptozotocin and liposomal doxorubicin is safe and efficient in patients with endocrine pancreatic tumors (EPTs). No cardiac toxicity was reported. BACKGROUND: The combination of streptozotocin and doxorubicin has been shown to be superior to streptozotocin and fluorouracil in the treatment of metastatic EPTs. However, the risk of cardiac toxicity from anthracyclins hampers the usefulness of the drug combination. Liposomal doxorubicin has a lower frequency of cardiac adverse events compared to doxorubicin. We wanted to assess the efficacy and safety of combined streptozotocin and liposomal doxorubicin in patients with metastatic EPTs. METHODS: Thirty patients with metastatic EPTs were recruited from three medical centers in Norway and Sweden during a time period of 3 years. All patients had histopathologically confirmed diagnoses and bidimensionally measurable lesions. 30 mg/m(2) of liposomal doxorubicin was administered on day 1 of each cycle. During the first course, 1 g of streptozotocin was given on 5 consecutive days. Thereafter, 2 g of streptozotocin was given on day 1 only. Treatment was repeated every 3 weeks. RESULTS: Twelve of 30 patients (40%) achieved an objective radiological response with a median duration of 9 months. Stabilization of disease was achieved in 17 of 30 patients (57%) for a median duration of 11 months. Only one patient had progressive disease as best response. The 2-year progression-free survival was 18% and the 2-year overall survival was 72%. The treatment was well tolerated. None of the patients experienced cardiac toxicity. CONCLUSION: We conclude that combined streptozotocin and liposomal doxorubicin is a safe and efficient treatment for EPTs. The efficacy seems to be comparable to that of combined streptozotocin and doxorubicin, whereas the cardiac toxicity clearly favors using the liposomal drug combination. PMID- 18285679 TI - Responses of the hypothalamopituitary adrenal axis and the sympathoadrenal system to isolation/restraint stress in sheep of different adiposity. AB - There is evidence that levels of adipose tissue can influence responses of the hypothalamopituitary-adrenal (HPA) axis to stress in humans and rats but this has not been explored in sheep. Also, little is known about the sympathoadrenal responses to stress in individuals with relatively different levels of adipose tissue. We tested the hypothesis that the stress-induced activation of the HPA axis and sympathoadrenal system is lower in ovariectomized ewes with low levels of body fat (lean) than ovariectomized ewes with high levels of body fat (fat). Ewes underwent dietary manipulation for 3 months to yield a group of lean ewes (n = 7) with a mean (+/-SEM) live weight of 39.1 +/- 0.9 kg and body fat of 8.9 +/- 0.6% and fat ewes (n = 7) with a mean (+/-SEM) live weight of 69.0 +/- 1.8 kg and body fat of 31.7 +/- 3.4%. Fat ewes also had higher circulating concentrations of leptin than lean ewes. Blood samples were collected every 15 min over 8 h when no stress was imposed (control day) and on a separate day when 4 h of isolation/restraint was imposed after 4 h of pretreatment sampling (stress day). Plasma concentrations of adrenocorticotropic hormone (ACTH), cortisol, epinephrine and norepinephrine did not change significantly over the control day and did not differ between lean and fat ewes. Stress did not affect plasma leptin levels. All stress hormones increased significantly during isolation/restraint stress. The ACTH, cortisol and epinephrine responses were greater in fat ewes than lean ewes but norepinephrine responses were similar. Our results suggest that relative levels of adipose tissue influence the stress-induced activity of the hypothalamopituitary-adrenal axis and some aspects of the sympathoadrenal system with fat animals having higher responses than lean animals. PMID- 18285680 TI - Neuropeptide W influences the excitability of neurons in the rat hypothalamic arcuate nucleus. AB - Neuropeptide W (NPW) is a ligand of the recently deorphaned receptor GPR7. Intracerebroventricular injection of this peptide results in reduced serum growth hormone concentration. Using whole-cell patch clamp recordings from somatostatin (SS) neurons in the hypothalamic arcuate nucleus, identified post-hoc using single-cell RT-PCR, we investigated the effects of NPW on membrane excitability. NPW application in acute slices of the arcuate nucleus resulted in the depolarization of the majority (62.5%) of the SS neurons tested, while smaller proportions of cells showed hyperpolarization or no response. Both the depolarization and hyperpolarization of arcuate SS neurons were preserved during recordings where voltage-gated sodium channels were blocked with tetrodotoxin, suggesting direct effects of NPW on the excitability of SS neurons. The observed depolarization of the majority of the SS neurons tested suggests that the central effects of NPW to inhibit growth hormone release results from activation of arcuate SS neurons, which could result in an inhibition of GHRH-releasing neurons. PMID- 18285681 TI - Cognitive status in hypothyroid female patients: event-related evoked potential study. AB - AIM: Higher brain functions are adversely affected in hypothyroid patients. The central nervous system features of hypothyroidism include mental retardation, various cognitive and memory deficits that have been evaluated by different neuropsychological tests. In the present study event-related potentials (ERPs) have been used to assess the cognitive status of hypothyroid patients before and after treatment. METHODS: Twenty-six newly diagnosed hypothyroid female patients with a mean age of 31.85 +/- 8.57 years and 26 female controls with a mean age of 30.92 +/- 7.24 years were the subjects of the study. The ERPs were recorded on a computerized evoked potential recorder using the 10-20 system of electrode placement and the standard auditory 'oddball' paradigm. The first recording of ERPs was done at the time of diagnosis and the second recording was done 3 months after the start of treatment in a euthyroid state. RESULTS: Prolongation of latencies was found in the early ERP components (N100, P200 and N200) in hypothyroid patients compared to controls, while no significant group differences were found on P300. In the hypothyroid group all the ERP latencies decreased significantly after attainment of euthyroidism compared to pretreatment values. ERP amplitudes revealed no significant group differences between hypothyroid patients and controls, but significant amplitude increases were found on the waves of N200 and P300 after treatment compared with pretreatment values in the hypothyroid group. CONCLUSIONS: Hypothyroid patients showed prolongation of latencies only in the early ERP components compared to controls. In hypothyroid patients, ERP findings indicate a speeding of sensory and cognitive processing in a state of euthyroidism compared to the pretreatment condition. PMID- 18285682 TI - Do stem cells exist in the adult kidney? AB - Adult stem cells exist in many organs and play a critical role in normal cell turnover and the response to injury. The existence of adult stem cells in the mammalian kidney remains controversial. Kidney stem cells have been isolated and characterized by many groups, often with discrepant results. This article will review the current state of knowledge regarding adult kidney stem cells and discuss future directions for kidney stem cell research. PMID- 18285683 TI - Lowering of proteinuria in response to antihypertensive therapy predicts improved renal function in late but not in early diabetic nephropathy: a pooled analysis. AB - In late diabetic nephropathy (DN) the initial lowering of albumin excretion rate (AER) with antihypertensive therapy is proportional to the degree of subsequent preservation of glomerular filtration rate (GFR). Whether a similar relationship exists between AER and GFR in early diabetes is not known. The present analysis has compared AER and GFR responses to antihypertensive therapy in 33 published studies (77 treatment groups) of early and late DN in type 1 (T1) and type 2 (T2) diabetes, analyzed on an intention-to-treat basis. Prospective trials were included if the initial change in AER during the first year of therapy and the change in GFR during at least 2 years of follow-up could be estimated from group mean data. The initial % decreases in AER were 5.9 +/- 4.3 (T1), 10.5 +/- 5.4 (T2, normotensive) and 18.4 +/- 6.2 (T2, hypertensive) in early DN and 7.6 +/- 11.1 (T1) and 20.8 +/- 5.5 (T2) in late DN. The corresponding annual % rates of decline of GFR were 2.0 +/- 0.5 (T1), 1.6 +/- 0.5 (T2, normotensive) and 2.1 +/- 0.3 (T2, hypertensive) in early DN and 9.8 +/- 1.5 (T1) and 9.2 +/- 1.1 (T2) in late DN. AER and GFR responses in each treatment group were closely correlated in late nephropathy (T1, r = -0.67, p = 0.03; T2, r = 0.57, p = 0.02) but not in early nephropathy. In contrast to late DN, the initial decrease in AER with antihypertensive therapy was not shown to predict the subsequent rate of decline of GFR in early DN. It follows that assessment of renoprotection during antihypertensive therapy in early nephropathy should be based not only on albuminuria but also on the GFR response. PMID- 18285684 TI - Hepatitis C in hemodialysis patients: current global magnitude, natural history, diagnostic difficulties, and preventive measures. AB - Hepatitis C virus (HCV) infection is a significant cause of morbidity and mortality in hemodialysis (HD) patients. The reported prevalence of HCV among the HD population has varied greatly from 1.9 to 84.6% in different countries in recent years. The length of time on HD is generally believed to be associated with HCV acquisition in HD subjects. Nevertheless, several recent reports failed to recognize any significant role of blood transfusion. Although there are some considerations about the accuracy of serologic testing in detecting HCV in HD patients, the accumulated data in this review suggest the false-negativity rate to be not more than 1.66% (153/9,220). Therefore, substituting virologic for serologic testing in the routine diagnosis of HCV infection in HD patients seems unreasonable. Several phylogenetic analyzes of viral isolates suggested nosocomial patient-to-patient transmission of HCV among HD patients for which the main potential source is believed to be contaminated hands and articles. However, isolation of HCV-infected HD patients and use of dedicated machines are currently unjustified while strict adherence to universal precautions seems to be enough to control disease spread in HD units. The present article is an update on epidemiological and clinical features of HCV in HD population. PMID- 18285685 TI - Clinical implications of seasonal variations in hemodialysis patients. PMID- 18285686 TI - Vitiligo after hematopoietic cell transplantation: six cases and review of the literature. AB - AIM: To investigate the prevalence and clinical characteristics of vitiligo after allogeneic hematopoietic cell transplantation (AHCT). METHODS: The development of vitiligo was analyzed among 421 patients who underwent AHCT in Ibni Sina Hospital (University of Ankara) between 1988 and 2004. RESULTS: Among 421 patients, we describe 6 with generalized vitiligo occurring after AHCT for chronic myelogenous leukemia. Five of them had severe chronic graft-versus-host disease (GVHD). Vitiligo was accompanied by alopecia areata and acquired ichthyosis in 2 patients with GVHD. CONCLUSION: Melanocyte destruction caused by the autoimmune reactions triggered by chronic GVHD as well as a genetic predisposition might have played a role in the development of vitiligo in our patients. These data support the hypothesis that vitiligo is an autoimmune entity. PMID- 18285687 TI - Etanercept at different dosages in the treatment of generalized pustular psoriasis: a case series. AB - BACKGROUND: Generalized pustular psoriasis (GPP) is a severe and disabling variant of psoriasis. The treatment of GPP is challenging, often characterized by side effects or unsatisfactory response. Etanercept is a tumor necrosis factor alpha blocking agent that demonstrated a consistent efficacy in the control of psoriasis. OBJECTIVES: We aimed to evaluate the efficacy and safety profile of etanercept at different dosages in GPP. METHODS: Six patients affected by GPP, unresponsive to conventional treatment, received etanercept subcutaneously at the dosages of 25 and 50 mg biweekly for 48 weeks. RESULTS: Our experience led to the observation that the administration of etanercept 50 mg biweekly is an effective dosage, characterized by good efficacy and rapidity of effect. Patients who were continuously treated at this dosage for 24 weeks presented stable conditions and long-term maintenance until week 48 even after a dose reduction to 25 mg. CONCLUSION: We demonstrated a good and durable efficacy of etanercept in patients affected by GPP. PMID- 18285688 TI - Genital HPV lesions and molluscum contagiosum occurring in patients receiving anti-TNF-alpha therapy. PMID- 18285689 TI - Residual perception of moving objects: dissociation of moving and static objects in a case of posterior cortical atrophy. AB - BACKGROUND/AIMS: Several studies have reported specific cognitive deficits of posterior cortical atrophy (PCA) starting in the early stage; however, these characteristics have not been demonstrated as arising in the middle or late stage of the disease. We report a patient with PCA who exhibited atypical symptoms after the middle stage. METHODS: We observed and examined the patient for more than 10 years in clinical and everyday situations. RESULTS: With increasing symptoms, the patient gradually showed blind-like behavior. He could not find a static object even when it was in front of him, but he could see and catch a tossed ball very well. In addition, he could walk and pick up scattered golf balls without colliding with obstacles. Magnetic resonance imaging and single photon emission computed tomography indicated severe degeneration in the white matter of the parietal lobe and lateral occipital region, whereas the primary visual cortex was relatively intact. CONCLUSION: Several studies have reported specific cognitive deficits in PCA; however, the present results support the conclusion that not only the initial stage but also the middle and late stages of the disease have the potential to disturb specific cognitive systems. PMID- 18285690 TI - Creation of ventricular septal defects on the beating heart in a new pig model. AB - BACKGROUND/AIMS: So far, surgical and interventional therapies for muscular ventricular septal defects (mVSDs) beyond the moderator band have had their limitations. Thus, alternative therapeutic strategies should be developed. We present a new animal model for the evaluation of such strategies. METHODS: In a pig model (n = 9), anterolateral thoracotomy was performed for exposure of the left ventricle. mVSDs were created under two- and three-dimensional echocardiography with a 7.5-mm sharp punch instrument, which was forwarded via a left ventricular puncture without extracorporeal circulation. RESULTS: Creation of mVSDs was successful in all animals (n = 9) confirmed by echocardiography, hemodynamic measurements and autopsy. The defects were located in the midmuscular (n = 4), apical (n = 1), inlet (n = 2) and anterior part (n = 2) of the muscular septum. All animals were hemodynamically stable for further procedures. The diameter and shunt volume of the mVSDs were 4.8-7.3 mm (mean: 5.9 mm) and 12.9 41.3% (mean: 22.1%), respectively. Autopsy confirmed in all animals the creation of a substantial defect. CONCLUSION: The described new technique for creation of an mVSD on the beating heart in a pig model is suitable for the evaluation of new therapeutic strategies for mVSD closure. PMID- 18285691 TI - Transcription of Corynebacterium glutamicum genes involved in tricarboxylic acid cycle and glyoxylate cycle. AB - Transcription of the tricarboxylic acid cycle genes of Corynebacterium glutamicum was investigated. Northern hybridizations revealed that gltA-fkb, odhA-orfA, sucC sucD, sdhC-sdhA-sdhB-orfB and mdh-orfC were transcribed as polycistronic mRNAs of size 1.9, 4.5, 2.5, 4.0 and 1.7 kb, respectively. The acn-acnR-gat gene cluster was transcribed as a mono-, bi- or tricistronic mRNA, depending on the carbon source. The 2.9-kb (acn) and 1.5-kb (acnR-gat) mRNAs, which were regulated by different promoters upstream of acn and acnR, were inversely expressed in acetate and glucose. The 4.5 kb (acn-acnR-gat) mRNA was constitutively expressed. The sizes of the mRNAs were 2.3, 2.1, 1.5, 1.3, 1.7, 1.5 and 2.9 kb for icd, sucB, fum, mdhB, mqo, aceA and aceB, respectively, indicating monocistronic transcription of these genes. RNA ligase-mediated rapid amplification of cDNA ends analysis of C. glutamicum RNA showed that the transcriptional start sites of gltA, acn, icd, odhA, sucB, sucC, sdhC, fum, mdh, mdhB, mqo, aceA and aceB were located 121, 107, 31, 99, 46, 83, 15, 25, 33, 23, 70, 111 and 183 bp upstream from the first nucleotide of the respective translation initiation codons. Alignment of these gene promoter regions provided evidence for highly conserved sequences that exhibited similarity to the sigma(A) consensus promoter sequences of Gram-positive bacteria. PMID- 18285692 TI - Protective role of CYP1A1*2A in the development of multiple myeloma. AB - We had previously reported the association of the NQO1*2/*2 polymorphism with a decreased risk for multiple myeloma (MM) in Koreans (odds ratio, OR, 0.24; 95% confidence interval, CI, 0.01-0.68). The associations of polymorphisms of other metabolizing enzymes (CYP1A1, GSTM1 and GSTT1) with the MM risk were investigated in 116 Korean MM patients and 176 Korean controls using TaqMan allelic discrimination and multiplex polymerase chain reaction. The ORs for CYP1A1*1/*2A and CYP1A1*1/*2B genotypes were 0.43 (95% CI, 0.19-0.98) and 0.51 (95% CI, 0.26 0.98), respectively, which was significantly associated with a decreased MM risk. With regard to CYP1A1 alleles, the OR for the CYP1A1*2A allele was 0.57 (95% CI, 0.326-0.995), which was also significantly associated with a decreased MM risk. However, null types of GSTM1 and GSTT1 polymorphisms were not associated with the MM risk. These results were different from those of a previous report on Caucasians which suggested the association of the GSTT1 polymorphism with an increased MM risk and no association of CYP1A1 with the MM risk. The associations of polymorphisms of metabolizing enzymes with the risk for MM differed between Koreans and Caucasians, suggesting an ethnic variation in the susceptibility to MM. PMID- 18285693 TI - t(16;21)(q24;q22) in acute myeloid leukemia: case report and review of the literature. PMID- 18285694 TI - Spontaneous resolution of severe aplastic anemia following thymic hemorrhage. AB - Over the last 7 years we have seen more than 200 severe aplastic anemia patients at this centre. Three of them developed an unusual complication in the form of thymic hemorrhage. Following this complication, all 3 patients recovered partially from their aplastic anemia, without any need for further immunosuppression. These cases show possible ways to manipulate the thymus gland as a management strategy for this disease. PMID- 18285695 TI - A comprehensive review of acute promyelocytic leukemia in children. AB - The outcome of patients with acute promyelocytic leukemia (APL) has substantially improved since the successful introduction of tretinoin, and nowadays combining tretinoin with chemotherapy is potentially curative for at least 70-75% of patients with newly diagnosed APL. In most pediatric series, APL represents < or = 10% of childhood acute myelogenous leukemia. APL in children is more common in girls and in obese children. It is characterized by a higher incidence of hyperleukocytosis, an increased incidence of microgranular morphology and by more frequent occurrence of the PML/RARalpha isoforms bcr 2 and bcr 3 compared to adults. Tretinoin-based therapy is curative for the majority of children with APL. Recent data indicate that > or = 2 negative RT-PCR assays for PML/RARalpha on bone marrow performed at least 1 month apart after completing therapy are strongly associated with long-term remissions, while conversion to PCR positivity for PML/RARalpha during remission is highly predictive of impending relapse. Data from recent studies in adults and limited data from children show that arsenic trioxide is the single most effective agent in APL and deserves immediate study in newly diagnosed children in an effort to further improve prognosis and to limit exposure to conventional cytotoxic chemotherapy. PMID- 18285696 TI - Interleukin-6 is a better predictor of mortality as compared to C-reactive protein, homocysteine, pentosidine and advanced oxidation protein products in hemodialysis patients. AB - Inflammatory markers predict mortality in hemodialysis (HD) patients, whereas a possible association between oxidative stress (OS) markers and survival is less clear. We assessed the impact on all-cause mortality of baseline inflammatory [high-sensitivity C-reactive protein and interleukin-6 (IL-6)] and OS markers (advanced oxidation protein products, pentosidine, homocysteine) in 112 HD patients. We found no significant correlations between inflammatory and OS markers. During the 5.5 years of follow-up, 51 patients died. In a Kaplan-Meier analysis, the survival rate was reduced in patients with IL-6 higher than the median (IL-6 >4.2 pg/ml) (log- rank = 6.47; p = 0.01), in diabetics (log-rank = 12.26; p = 0.0005) and in older patients (log-rank = 11.22; p = 0.0008). Moreover, in Cox analysis only IL-6 and age were independently associated with mortality. We conclude that in this group of prevalent Brazilian HD patients, IL 6 was a better predictor of survival than other inflammatory and OS markers. PMID- 18285697 TI - High time for high-flux hemodialysis mechanistic studies. PMID- 18285698 TI - Effects of low- and high-flux dialyzers on oxidative stress and insulin resistance. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of mortality in patients with end-stage renal disease (ESRD). The cornerstone of high CVD incidence in ESRD patients is endothelial dysfunction which results from inflammation, oxidative stress and insulin resistance. Although various modalities of hemodialysis (HD) have been presumed to exert different effects on oxidative stress and insulin resistance, solid evidence is still lacking. METHODS: 40 ESRD patients undergoing HD were prospectively enrolled and divided randomly into two groups. Patients in each group received either F8 HPS (low flux) (Group A) or FX80 (high-flux) (Group B) as HD dialyzers for 2 consecutive months. Diet pattern and medications were kept as usual in both groups to avoid considerable blood glucose change during study period. Blood samples were taken at the start and end of the study. RESULTS: A total of 38 patients (18 and 20 for Groups A and B, respectively) completed the study. Within each group, there was no change in adiponectin, plasma 8-iso-prostaglandin F(2)(alpha), high sensitivity C-reactive protein, blood glucose and insulin after 2 months of treatment except a significant change of HOMA(IR) (p = 0.02) in high-flux group. The significant change of HOMA(IR) between the two groups (p = 0.017) mainly results from the parallel change of insulin between the two groups (p = 0.03). CONCLUSION: For patients receiving HD, the high-flux dialyzer with synthetic polysulfone membranes fails to provide a better anti-inflammatory or antioxidative effect than the low-flux dialyzer; however, the high-flux dialyzer does significantly improve insulin resistance in this short-term study. This result implies that the high-flux dialyzer might provide better cardiovascular protection than the low-flux dialyzer. Therefore, the low-flux dialyzer might be considered for patients who only need short-term HD therapy. Regarding patients under long-term maintenance HD therapy, a high-flux dialyzer might be the choice of dialyzer. PMID- 18285699 TI - SNF4Agamma, the Drosophila AMPK gamma subunit is required for regulation of developmental and stress-induced autophagy. AB - In holometabolous insects including Drosophila melanogaster a wave of autophagy triggered by 20-hydroxyecdysone is observed in the larval tissues during the third larval stage of metamorphosis. We used this model system to study the genetic regulation of autophagy. We performed a genetic screen to select P element insertions that affect autophagy in the larval fat body. Light and electron microscopy of one of the isolated mutants (l(3)S005042) revealed the absence of autophagic vesicles in their fat body cells during the third larval stage. We show that formation of autophagic vesicles cannot be induced by 20 hydroxyecdysone in the tissues of mutant flies and represent evidence demonstrating that the failure to form autophagic vesicles is due to the insertion of a P-element into the gene coding SNF4Agamma, the Drosophila homologue of the AMPK (AMP-activated protein kinase) gamma subunit. The ability to form autophagic vesicles (wild-type phenotype) can be restored by remobilization of the P-element in the mutant. Silencing of SNF4Agamma by RNAi suppresses autophagic vesicle formation in wild-type flies. We raised an antibody against SNF4Agamma and showed that this gene product is constitutively present in the wild-type larval tissues during postembryonal development. SNF4Agamma is nearly absent from the cells of homozygous mutants. SNF4Agamma translocates into the nuclei of fat body cells at the onset of the wandering stage concurrently with the beginning of the autophagic process. Our results demonstrate that SNF4Agamma has an essential role in the regulation of autophagy in Drosophila larval fat body cells. PMID- 18285700 TI - Emodin enhances cytotoxicity of chemotherapeutic drugs in prostate cancer cells: the mechanisms involve ROS-mediated suppression of multidrug resistance and hypoxia inducible factor-1. AB - The intrinsic or acquired resistance to multiple drugs (MDR) of cancer cells remains one of the main obstacles for chemotherapy. Development of small molecule targeting to hypoxia inducible factor-1 (HIF-1) has been recently proposed as strategy for treatments of drug-resistant solid tumors. In the present study, emodin, proven as a reactive oxygen species (ROS) generator by our previous work, was applied in combination with cisplatin and other chemotherapeutic drugs in the multidrug resistant prostate carcinoma cell line DU-145 and normal human dermal fibroblasts. Results showed that emodin/cisplatin co-treatment remarkably elevated ROS level and enhanced chemosensitivity in DU-145 cells, compared with cisplatin-only treatment, but exerted little effect on non-tumor cells. The effect of co-treatment on MDR1 gene and its upstream regulator HIF-1 was then investigated in DU-145. Co-treatment downregulated MDR1 expression and promoted drug retention, and meanwhile suppressed transactivation of HIF-1 in response to hypoxia without changing expression of HIF-1 alpha. The experiments on tumor bearing mice showed that co-treatment inhibited the tumor growth in vivo, owing to oxidative stress and MDR1 down-regulation within tumors. HIF-1 transactivation and clonegenesis were suppressed in cells isolated from the tumors. Finally, examinations for the body weight, the organ histology and the antioxidant capacity of serum suggested that no systemic toxicity related to co-treatment was discernable. In conclusions, emodin, as a novel small inhibitor of HIF-1, may be recognized an effective adjunctive to improve efficacy of cytotoxic drugs in prostate cancer cells with over-activated HIF-1 and potent MDR. PMID- 18285701 TI - HuRrying colon cancer progression. PMID- 18285702 TI - Jun activation domain-binding protein 1 negatively regulate p27 kip1 in non Hodgkin's lymphomas. AB - OBJECTIVE: Recent evidences suggest that Jun activation domain-binding protein 1 (Jab1) specifically interacts with the Cdk inhibitor p27kip1 and induces nuclear export and subsequent degradation of p27kip1. The purpose of this study is to investigate whether Jab1 expression is correlated with p27kip1 level in Non Hodgkin's Lymphomas(NHLs) and how it influenced the stability of p27kip1, as well as whether Jab1 expression is associated with clinicopathologic variables and prognosis of NHLs. RESULTS: Immunohistochemical analysis showed that Jab1 expression was negatively associated with p27kip1 level and significantly associated with unfavorable clinicopathologic variables. Overexpression Jab1 in lymphoma cell lines Jurkat resulted in decreased p27kip1 level and advanced progression from G(1) to S phase of the cell cycle. Subcellular fractionation confirmed Jab1 could lead to nuclear export of p27kip1. Phosphorylation of p27kip1 at Ser10 and Thr157 was significantly increased after Jab1 transient transfected, while Thr187 phosphorylation was decreased. Immunoprecipitation demonstrated that Jab1 overexpression facilitated p27kip1 to dissociate from Cdk2 and associated with Cdk4. Finally, Survival analysis revealed that Jab1 overexpression is significantly associated with overall survival (P = 0.000). When Jab1 and p27kip1 are combined, patients with Jab1(+)/p27kip1(-) revealed poorer overall survival (p = 0.001). Multivariate analysis revealed that Jab1 protein is an independent prognostic indicator for overall survival. METHODS: Immunohistochemical and/or Western blot analysis was done in 116 cases of NHLs and Jurkat cells. CONCLUSIONS: These findings suggest that Jab1 protein may contribute to the tumor progression through Jab1-mediated p27kip1 degradation and that control of Jab1 expression is a novel therapeutic target with NHLs. PMID- 18285703 TI - Tumor-initiating stem cells in liver cancer. AB - It is widely accepted that cancer is a disease of stem cells. Definite evidence suggests that tumors harbor a small population of cancer stem cells (CSC) that both give rise to the bulk of the tumor and are tumorigenic in experimental models. Mounting evidence suggests that these cells are responsible for regrowth of a tumor following unsuccessful treatment and for the establishment of metastases. The concept of CSC has been demonstrated in several human cancers including leukemia, breast, prostate, lung, pancreas, colon and brain tumors. Recently, several studies have demonstrated that liver cancer, like other tumors, are derived from a small population of liver cancer stem/progenitor cells. Although still controversial, Liver CSC will likely become the most crucial target in the treatment of liver cancer, and a thorough understanding of its origin, molecular profile and particularly of how the Liver CSC differs from the normal stem cells, might allow it to be targeted selectively and eliminated, thus improving therapeutic outcome. In this review we will summarize the recent evidence for Liver CSC, the relationship between normal and liver stem cells, and the possibility that transformation of different cell types in the liver may generate different types of liver cancers. PMID- 18285704 TI - A comparative study of gene vaccines encoding different extracellular domains of the vascular endothelial growth factor receptor 2 in the mouse model of colon adenocarcinoma CT-26. AB - AIMS: To compare the immunogenicity and anti-tumor effects of gene vaccines encoding domains 1-4 with full length of extracellular region of VEGFR2. RESULTS: Both DNA vaccines decreased VEGF levels and rose specific antibodies; the lymphocyte subsets of vaccinated mice maintained high; tumor latency period and survival time of immunized mice were prolonged; tumor size, weight, MVD and liver metastases were significantly less than the control groups. METHODS: Mouse model of CT-26 adenocarcinoma of colon were treated with orally immunized gene vaccine encoding extracellular 1-4 and full length of VEGFR2 respectively. The effect of anti-tumor was evaluated by detecting the tumor volume, mice survival time, intratumoral microvessel density (MVD) and liver metastases. To explore the reasonable mechanism of the oral gene vaccines, the levels of VEGF and anti VEGFR2 antibody in serum were detected by ELISA, CD4+ and CD8+ T cells in peripheral blood and subcutaneous tumors were analyzed by flow cytometer and immunohischemistry respectively. CONCLUSION: Compared with full length of extracellular domain of VEGFR2, extracellular regions 1-4 of VEGFR2 has been sufficient to decrease the serum VEGF level and to inhibit tumor growth and metastasis specifically. PMID- 18285705 TI - Heterophilic NeuGcGM3 ganglioside cancer vaccine in advanced melanoma patients: results of a Phase Ib/IIa study. AB - NeuGcGM3 ganglioside is especially attractive because it is expressed on melanoma cells but it is minimally or not expressed at all on most normal human tissues. A Phase Ib/IIa clinical trial was carried out in patients with advanced cutaneous and ocular malignant melanomas, to evaluate immunogenicity and toxicity of an intramuscularly administered cancer vaccine and composed by NeuGcGM3 in a proteoliposome of Neisseria meningitides with Montanide ISA 51 as adjuvant. Twenty two patients were included, twelve at dose level of 200 microg and 10 at 400 microg. The first five doses were administered every other week and then monthly until 9 doses. 12 patients received additional immunizations. Vaccination induced specific anti-NeuGcGM3 IgM, IgG and IgA antibodies responses. Titers of IgM were greater for the highest vaccine doses. Vaccination also elicited DTH response in 45.5% of patients in the lower doses and 77.8% in the higher doses. Toxicities were mostly grade 1 or 2, according CTC-NCI criteria. Interestingly, 3 patients developed vitiligo at the lower dose (none in the highest dose) although the nominal antigen NeuGcGM3 is not present in melanocytes. Survival analysis was not the goal of this Phase I trial; nevertheless, the fact that seven patients are alive for more than 2 years after inclusion is noteworthy. Safety and immunogenicity with NeuGcGM3 vaccine treatment in advanced melanoma patients were established. The prognostic value of autoimmunity and the possibilities of dissociating anti-tumor immunity from autoimmunity deserve further research. PMID- 18285706 TI - Chemotherapy targeting regional lymphatic tissues to treat rabbits bearing VX2 tumor in the mammary glands. AB - Lymph node status is one of the decisive prognostic factors in breast cancer. Chemotherapy targeting regional lymphatic tissues has emerged as a promising approach for the treatment of malignancies with high tendency to disseminate lymphatically. In this study, rabbits bearing well-developed VX2 tumors in the mammary gland received carboplatin-activated carbon suspension (CP-CH) administered subcutaneously (s.c.), carboplatin solution (CP-Sol) s.c., CP-Sol intravenously (i.v.), or a sham therapy, respectively, 6 weeks after tumor implantation. The platinum concentrations in breast tumors, axillary lymph nodes and distant metastases were determined by Zeeman atomic absorption spectrometry (AAS). Therapeutic effects were assessed in terms of tumor and node volume ratio, cell proliferation and necrosis. Compared to CP-CH or CP-Sol s.c., CP-Sol i.v. exhibited more marked cytotoxic efficacy on breast tumor and distant metastases. In contrast, CP-CH s.c. produced much higher drug concentrations and more evident antitumor activities in axillary nodes, as demonstrated by the inhibited node growth, declined mRNA level of proliferating cell nuclear antigen (PCNA), and more extensive tumor necrosis. Based on these data, we recommend lymphatic chemotherapy to be used as an effective component of the approach to the systemic treatment on breast cancer patients with positive lymph nodes. PMID- 18285707 TI - Validation and toxicity of PI3K/Akt pathway inhibition by HIV protease inhibitors in humans. AB - PURPOSE: Activation of the phosphatidylinositol 3-kinase/Akt pathway in tumors leads to radiation resistance, and inhibition of this pathway radiosensitizes tumors in laboratory models. Several first-generation human immunodeficiency virus (HIV) protease inhibitors (HPIs) inhibit Akt activation and are radiosensitizers. In order to validate a biomarker of Akt activity in anticipation of clinical trials using HPIs combined with radiotherapy, we sought to determine whether Akt activation was inhibited in leukocytes of HIV+ patients that were already taking these agents. RESULTS: Patients taking these "active" radiosensitizing protease inhibitors had low levels of phospho-Akt compared to HIV+ patients taking either no medications or other anti-retroviral regimens. We found no significant differences in acute toxicities or in the ability to finish radiation treatment between 14 patients taking radiosensitizing HPIs and the 28 controls. METHODS AND MATERIALS: Peripheral blood mononuclear cells from HIV+ patients either taking radiosensitizing HPIs (nelfinavir, saquinavir, amprenavir) or not were analyzed by Western blotting for phospho-Akt. In order to determine whether these radiosensitizing HPIs increase the toxicity of radiotherapy, we performed a retrospective cohort study of HIV+ cancer patients treated with radiation and compared patients on radiosensitizing HPIs to controls not taking these agents. CONCLUSIONS: These results demonstrate the proof of principle that HPIs can inhibit Akt activation in patients taking normally prescribed anti retroviral doses and are not associated with excessive toxicity. Radiosensitizing HPIs are excellent candidates for Phase I clinical trials as radiation sensitizers, and peripheral blood mononuclear cells can be used as a drug activity biomarker for Akt pathway inhibition. PMID- 18285708 TI - Effect of blocking VEGF, hTERT and Bcl-xl by multiple shRNA expression vectors on the human laryngeal squamous carcinoma xenograft in nude mice. AB - BACKGROUND: In our previous work, we constructed short hairpin RNA (shRNA) expression plasmids that targeted human telomerase reverse transcriptase (hTERT) messenger RNA, and we demonstrated that these vectors could inhibit the growth of cancer cells by 76.5% in vivo. In order to maximize the efficiency and versatility of the vector-based siRNA approach, here we have constructed multiple shRNA expression vectors that simultaneously targeted 3 different genes in cancer cells, and then investigated their effect on human laryngeal squamous carcinoma (Hep-2) in vivo. RESULTS: We found that expression of multiple shRNAs led to a significant reduction in VEGF, hTERT and Bcl-xl mRNA and protein expression. Tumor growth curves showed that those tumors treated with the shRNAs were obviously smaller than control, nontransfected tumors. Analysis at 14 days following the final injection showed a tumor inhibition ratio of 91.2%. However, the control shRNA vectors showed none of these effects. MATERIALS AND METHODS: Short hairpin RNA expression vectors targeting the mRNA of VEGF, hTERT and Bcl-xl were constructed and subsequently transfected by direct injections into the tumors formed by Hep-2 cells implanted in nude mice. The expression of targeted genes as well as apoptosis of tumor cells were evaluated after treatment with multiple shRNA vectors or control vectors. CONCLUSIONS: Our results demonstrate that the application of vector-based RNAi technology that involves blocking multiple targets will be a promising therapeutic modality in the gene therapy of human cancers. The data strongly suggest that simultaneously blocking multiple genes in human cancers using an RNAi approach should be considered in cancer therapy. PMID- 18285709 TI - Immunologic, virologic, and clinical consequences of episodes of transient viremia during suppressive combination antiretroviral therapy. AB - OBJECTIVE: To investigate immunologic, virologic, and clinical consequences of episodes of transient viremia in patients with sustained virologic suppression. METHODS: From the AIDS Therapy Evaluation Project, Netherlands cohort, 4447 previously therapy-naive patients were selected who were on continuous combination antiretroviral therapy and had initial success (2 consecutive HIV RNA measurements <50 copies/mL). During episodes of viral suppression (RNA <50 copies/mL), low-level viremia (RNA 50 to 1000 copies/mL), or high-level viremia (RNA >1000 copies/mL) after initial success, the occurrence of therapy changes, drug resistance, and clinical events was assessed. RESULTS: During 11,187 person years of follow-up, 1281 (28.8%) patients had at least 1 RNA measurement >50 copies/mL. Among 8069 episodes, there were 5989 (74.2%) episodes of suppression, 1711 (21.2%) episodes of low-level viremia, and 369 (4.6%) episodes of high-level viremia. Most episodes of low-level viremia consisted of < or =2 RNA measurements (93.7%), were without clinical events or therapy changes (79.6%), and were without changes in CD4 cell counts. Therapy changes (52.3% of episodes) and resistance (23.3%) were frequently observed during high-level viremia. CONCLUSIONS: Episodes of low-level viremia are frequent and short lasting, and the low proportion of episodes with clinical events suggests that leaving therapy unchanged is a clinically acceptable strategy. In contrast, high-level viremia is associated with resistance and is often followed by therapy changes. PMID- 18285710 TI - Impact on hemoglobin of starting combination antiretroviral therapy with or without zidovudine in anemic HIV-infected patients. AB - OBJECTIVE: To evaluate, among anemic patients with HIV, the impact on hemoglobin (Hb) of initiating zidovudine (AZT)-containing and non-AZT-containing combination antiretroviral therapy (cART). METHODS: We used medical records data collected in 11 US cities from 1998 to 2004. Baseline anemia was described as mild (10 < Hb < or = 12 [women] or 14 [men] g/dL), moderate (8 < Hb < or = 10 g/dL), or severe (Hb < or = 8 g/dL). Improvement of anemia was a > or =1-g/dL increase in Hb, with a decrease in categoric severity. We excluded patients previously treated with erythropoietin or transfusion, and used Cox proportional hazards regression to describe factors associated with hazard of improvement of anemia. RESULTS: For 1620 patients initiating cART, more than half (54%) of patients had improvement of anemia. Time to improvement of anemia was longer for those initiating AZT containing regimens and blacks and was shorter for those with moderate and severe anemia or CD4 counts <200 cells/microL. CONCLUSIONS: Most anemic patients initiating cART (with or without AZT) had increases in Hb-especially those with more severe anemia or immunosuppression. Initiation of AZT-containing cART may be considered, even for patients with preexisting anemia; however, improvement of anemia may be delayed for black patients and for patients with mild disease. PMID- 18285711 TI - Regional adipose tissue and elevations in serum aminotransferases in HIV-infected individuals. AB - BACKGROUND: The association of fat distribution with alanine aminotransferase (ALT) and aspartate aminotransferase (AST) elevations is not well-defined in HIV infected individuals. Obesity is associated with hepatic steatosis, and ALT is a marker of steatosis in the general population. METHODS: Cross-sectional analysis of 1119 HIV-infected and 284 control subjects. Hepatitis C virus (HCV) RNA testing determined HCV infection. Magnetic resonance imaging measured regional adipose tissue volume. RESULTS: After adjustment for demographic and lifestyle factors, visceral adipose tissue (VAT) was positively associated with ALT in HIV/HCV-coinfected subjects (+9.8%, 95% confidence interval [CI]: 2.8 to 17.6), HIV-monoinfected subjects (+8.0%, 95% CI: 4.2 to 12.1), and controls (+5.9%, 95% CI: 2.0 to 10.1). In contrast, lower trunk subcutaneous adipose tissue (SAT) was negatively associated with ALT in HIV/HCV-coinfected subjects (-14.3%, 95% CI: 24.7 to -4.2) and HIV-monoinfected subjects (-11.9%, 95% CI: -18.4 to -5.3); there was a trend toward an association in controls (-7.1%, 95% CI: -22.7 to 5.9). Estimated associations between regional adipose tissue and AST were small and did not reach statistical significance. CONCLUSIONS: More VAT and less lower trunk SAT are associated with elevated ALT, which likely reflects the presence of steatosis. There was little association with AST. HCV infection and having more VAT or less lower trunk SAT are independently associated with elevated ALT in HIV infection. Study regarding the association between VAT, trunk SAT, HCV, and progression of steatosis and fibrosis is needed in HIV-infected individuals. PMID- 18285712 TI - Effect of trimethoprim-sulfamethoxazole prophylaxis on antimicrobial resistance of fecal Escherichia coli in HIV-infected patients in Tanzania. AB - BACKGROUND: Trimethoprim-sulfamethoxazole (SXT) reduces morbidity and mortality among HIV-infected persons in Africa, but its impact on antimicrobial resistance is of concern. METHODS: HIV-uninfected (group A), HIV-infected but not requiring SXT (group B), and HIV-infected and eligible for SXT (group C) adults were recruited into a prospective observational cohort study in Moshi, Tanzania. Stool was examined for Escherichia coli nonsusceptible to SXT at baseline and at weeks 1, 2, 4, and 24. General estimating equation models were used to assess differences in susceptibility over time and cross-resistance to other antimicrobials. RESULTS: Of 181 subjects, 118 (65.1%) were female and the median (range) age was 36 (20 to 72) years. At baseline, E. coli nonsusceptible to SXT was isolated from 23 (53.5%) of 43 patients in group A, 25 (67.6%) of 37 patients in group B, and 37 (64.9%) of 57 patients in group C. The odds ratios (P value) for SXT nonsusceptibility in group C at weeks 1, 2, 4, and 24 compared with baseline were 3.4 (0.013), 3.0 (0.019), 2.9 (0.030), and 1.5 (0.515), respectively. SXT nonsusceptibility was associated with nonsusceptibility to ampicillin, chloramphenicol, ciprofloxacin, and nalidixic acid (P or=50 cells/microL and achievement of undetectable plasma HIV RNA level), 15.8% an immunologic response only (CD4+/VL(-)), 8.7% a virologic response only (CD4(-)/VL+), and 5.0% a concordant unfavorable response (CD4( )/VL(-)). Both types of discordant responses (CD4+/VL(-) and CD4(-)/VL+), nonresponse (CD4(-)/VL(-)), and baseline CD4 cell count were significantly associated with earlier development of an OI or death (relative hazard [RH] = 2.81, 95% confidence interval [CI]: 1.31 to 3.97; RH = 4.83, 95% CI: 2.10 to 11.12; and RH = 0.93, 95% CI: 0.88 to 0.99, respectively). CD4+/VL(-) and CD4( )/VL(-) were associated with nonwhite race in multivariate logistic regression models (adjusted OR = 2.83, 95% CI: 1.46 to 5.47 and adjusted OR = 6.50, 95% CI: 1.65 to 25.69, respectively). CONCLUSION: Discordant immunologic and virologic responses at 3 to 9 months after HAART initiation play important roles in predicting long-term clinical outcomes in treatment-naive patients. PMID- 18285714 TI - Brief strengths-based case management promotes entry into HIV medical care: results of the antiretroviral treatment access study-II. AB - OBJECTIVE: The Antiretroviral Treatment Access Study-II (ARTAS-II) evaluated a brief case management intervention delivered in health departments and community based organizations (CBOs) to link recently diagnosed HIV-infected persons to medical care rapidly. METHODS: Recently diagnosed HIV-infected persons were recruited from 10 study sites across the United States during 2005 to 2006. The intervention consisted of up to 5 sessions with an ARTAS linkage case manager over a 90-day period. The outcome measure was whether or not the participant had seen an HIV medical care provider at least once within 6 months of enrollment. Multivariate logistic regression was used to identify significant predictors of receiving HIV medical care. RESULTS: Seventy-nine percent (497 of 626) of participants visited an HIV clinician at least once within the first 6 months. Participants who were older than 25 years of age, Hispanic, and stably housed; had not recently used noninjection drugs; had attended 2 or more sessions with the case manager; and were recruited at a study site that had HIV medical care colocated on its premises were all significantly more likely to have received HIV care. CONCLUSIONS: The ARTAS linkage case management intervention provides a model that health departments and CBOs can use to ensure that recently diagnosed HIV-infected persons attend an initial HIV care encounter. PMID- 18285715 TI - Antiretroviral drug resistance and risk behavior among recently HIV-infected men who have sex with men. AB - OBJECTIVES: Examine associations among behaviors including substance use during sexual encounters, and transmitted HIV drug resistance in recently HIV-infected men who have sex with men (MSM). METHODS: Between 2002 and 2006, 117 recently HIV infected MSM completed questionnaires regarding their 3 most recent sexual partners. Serum samples were tested for the presence of genotypic and phenotypic HIV drug resistance. Logistic regression analysis was used to assess the association of substance use, behaviors, and resistance to at least 1 class of HIV drugs. RESULTS: The mean age of participants was 35 years; 71% identified as white and 19% as Hispanic. Sixty (51%) reported substance use during sexual activity in the past 12 months. A total of 12.5% of 112 had genotypic drug resistance to at least 1 class of antiretroviral medications, and 14% of 117 had phenotypic drug resistance. Substances used during sexual activity associated with phenotypic drug resistance in multivariate models included any substance use (adjusted odds ratio [aOR] = 4.21, 95% confidence interval [CI]: 1.13 to 15.68), polysubstance use (aOR = 5.64, 95% CI: 1.62 to 19.60), methamphetamine (aOR = 4.00, 95% CI: 1.19 to 13.38), 3,4-methylenedioxy-N-methylamphetamine (MDMA)/Ecstasy (aOR = 7.16, 95% CI: 1.40 to 36.59), and gamma-hydroxyl butyrate (GHB) (aOR = 6.98, 95% CI: 1.82 to 26.80). The genotype analysis was similar. CONCLUSIONS: Among these recently HIV-infected MSM, methamphetamine use during sexual activity and use of other substances, such as MDMA and GHB, was associated with acquired drug-resistant virus. No other behaviors associated with acquisition of drug-resistant HIV. PMID- 18285716 TI - Can we predict the degree of fibrosis in chronic hepatitis C patients using routine blood tests in our daily practice? AB - GOALS: To determine the validity of fibrosis indexes based on simple laboratory tests in daily practice. BACKGROUND: Fibrosis indexes were developed in referral centers using high-quality data. METHODS: We compared the performance characteristics of several such indexes with liver biopsies in a cohort of 490 diverse veterans with chronic hepatitis C from 24 centers. All laboratory tests including interpretation of the liver biopsy were done locally. The following indexes were calculated and correlated with a 5-point fibrosis stage (F0-F4) on liver biopsies: platelet counts (<100 or <150x10(9)/L), aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio (AAR), Pohl score, AST-to-platelet ratio index (APRI) and "Lok's model." RESULTS: Our cohort was predominantly male with 24% blacks, and fibrosis stages of 0, 1, 2, 3, and 4 in 11%, 24%, 28%, 24%, and 13%, respectively. All indexes performed better in predicting advanced (F3-4) than significant (F2-4) fibrosis. When patients with F3-4 were compared to those with F0-2, the area under the receiver operating characteristics curve were 0.534 and 0.641 for platelet count <100 and <150x10(9)/L, respectively, 0.524 for AAR, 0.534 for Pohl score, 0.693 for Lok's model, and 0.765 for APRI. The sensitivity, specificity, and predictive values of APRI and Lok's model were only slightly lower than those reported by the authors using the recommended cutoffs in clinical trial settings. Alcohol use within 12 months, normalization of AST, ALT, and race (blacks/non-blacks) had minimal impact on the performance. CONCLUSIONS: AAR, Pohl, and platelet counts <100x10(9)/L have limited ability to predict significant/advanced fibrosis with area under the receiver operating characteristics curve similar to 0.5. However, platelet counts <150x10(9)/L, Lok's model and APRI performed well for advanced fibrosis in our daily practice setting. PMID- 18285717 TI - Pegylated interferon alpha-2b plus ribavirin for naive patients with HCV-related cirrhosis. AB - BACKGROUND: Data on the efficacy of antiviral therapy in patients with HCV related compensated cirrhosis are generally drawn from analyzing subgroups in larger trials. AIMS: (1) To analyze the safety and efficacy of combination therapy in naive patients with HCV-related cirrhosis; (2) to evaluate the factors influencing the sustained virologic response (SVR) in cirrhotic patients by comparison with a group of noncirrhotic patients; (3) to analyze the outcome of cirrhotic patients either acquiring SVR and nonresponders to the antiviral therapy during the posttreatment follow-up. METHODS: We consecutively enrolled 365 patients with biopsy-proven HCV-related chronic hepatitis meeting the inclusion criteria for pegylated interferon a-2b plus Ribavirin: 87 patients had compensated liver cirrhosis and 278 had histologic stages between 1 and 4 according to Ishak's classification. RESULTS: The 2 groups were comparable for genotype, viral load, and alanine transferase at presentation. Cirrhotic patients were significantly older and had significantly higher body mass index, serum ferritin, and gamma-glutamyl transpeptidase. The rate of side effects was similar in the 2 groups, whereas the rate of SVR was significantly lower in cirrhotic (45.9%) than in noncirrhotic patients (65.8%). Logistic regression analysis showed that genotype 1 to 4 and high viral load were independent variables correlating with nonresponse in the sample as a whole. During follow-up, hepatocellular carcinoma developed in 5/38 (13.2%) cirrhotic patients not responding or relapsing after treatment. No cases of hepatocellular carcinoma were seen among cirrhotic or noncirrhotic patients with a SVR. CONCLUSIONS: Cirrhotic patients with compensated disease have a reasonably good chance of virologic response and should be offered treatment, carefully monitoring any side effects. PMID- 18285718 TI - Endoscopic sphincterotomy before deployment of covered metal stent is associated with greater complication rate: a prospective randomized control trial. AB - OBJECTIVE: Endoscopic sphincterotomy (ES) may facilitate insertion of self expandable metal stent (SEMS) and also help avert the development of pancreatitis from stent-related occlusion of the pancreatic duct. On the other hand, ES is also independently associated with pancreatitis, bleeding, and perforation. We evaluated whether ES before SEMS placement was associated with a greater likelihood of stent migration and other complications in patients with malignant obstruction of the distal common bile duct. METHODS: Seventy-four patients with unresectable distal bile duct obstruction were prospectively randomized to biliary stenting following ES (group 1) and without ES (group 2). Main outcome measures included overall procedure complications rates including stent migration, stent occlusion, oxygen desaturation, bleeding, perforation, and pancreatitis. RESULTS: Covered SEMS were successfully deployed in all patients in both groups. Stent migration occurred in 6 patients (16%) in group 1 and 1 patient (3%) in group 2, P=0.075. Overall, complications occurred in 18 patients in group 1 and 4 patients in group 2, P=0.006. There was no pancreatitis in either group. CONCLUSIONS: Deployment of covered SEMS without prior ES in patients with distal common bile duct obstruction owing to pancreatic cancer is feasible and prevents the development of complications such as stent migration, bleeding, and perforation. PMID- 18285719 TI - Health in young people: social inhibition and negative affect and their relationship with self-reported somatic complaints. AB - OBJECTIVE: It was investigated whether the combination of negative affect and social inhibition (personality Type D) is positively associated with somatic complaints in pre- and early adolescents. Moreover, an analysis of stability was made. METHODS: Data were collected twice, with an 18-month time interval, using questionnaires (N=668 children, M initial age=10.3, SD =.70). Individuals scoring above the median on social inhibition and negative affect were classified as Type D. These individuals had more somatic complaints than individuals scoring above the median on social inhibition or negative affect. Individuals with below median scores on social inhibition and negative affect had the lowest level of somatic complaints. RESULTS: The overall stability of the characteristics was moderate and most individuals did not show significant change on the two variables. CONCLUSIONS: Because the Type D classification was nevertheless not very stable, a different way of defining Type D is proposed. The social inhibition/negative affect combination indicates an elevated risk for self-reported somatic complaints in youth. PMID- 18285720 TI - Observational assessment of family functioning in families with children who have type 1 diabetes mellitus. AB - OBJECTIVE: Children with type 1 diabetes mellitus have a complex treatment regimen that includes insulin therapy and dietary requirements (e.g., matching insulin and carbohydrate intake). Previous research has shown that parents of children with type 1 diabetes mellitus report significant mealtime challenges and higher parenting stress compared to parents of healthy controls. The objective of the current study was to compare family functioning in children with type 1 diabetes mellitus (ages 2-8) to a matched, healthy control sample. Sixty-six families (33 diabetes; 33 controls) participated in a home visit at which their evening meal was videotaped. Tapes were then coded using the McMaster Interaction Coding System to objectively assess family functioning. RESULTS: Results indicated that families in the diabetes group demonstrated significantly poorer family functioning in a majority of areas (communication, affect management, family roles, overall functioning) compared to the healthy control sample. Additionally, families with lower socioeconomic status and families of male children evidenced poorer overall family functioning for both groups. CONCLUSIONS: Results also suggest that family-focused interventions for young children with type 1 diabetes should include components targeting family functioning in the areas of communication, affect management, and family roles. PMID- 18285721 TI - HIV/AIDS awareness building imperative. PMID- 18285722 TI - Fact of low hemoglobin and packed cell volume in pregnant women are at a stand still. AB - A cross sectional descriptive type of study was done in 98 women of reproductive age. Among them 25 were in control group of non pregnant women and 73 were pregnant women of 1st, 2nd and 3rd trimester of pregnancy with and without iron supplementation. The period of study was July 2004 to June 2005. The main objective of our study was to compare hemoglobin percent and packed cell volume (PCV) in pregnant and non pregnant women. In present study hemoglobin percent and packed cell volume was significantly decreased in 2nd and 3rd trimester of pregnancy when compared with the control group and same category of women who were not supplemented with iron. It is evident that the significantly low hemoglobin percent and packed cell volume (PCV) in pregnant women is due in part to dietary iron deficiency. Therefore, iron therapy in pregnancy is helpful to maintain the hemoglobin percent and packed cell volume nearer to that of non pregnant normal women. PMID- 18285723 TI - An anatomical study on pyramidal lobe of thyroid gland in Bangladeshi people. AB - The study was done to see the incidence of pyramidal lobe of thyroid gland including its position, attachment, length and incidence of its association with levator glandulae thyroidae in Bangladeshi population. Sixty postmortem tissue blocks containing thyroid gland and its related neighbouring structures were collected from 48 male and 12 female cadavers of different age groups and fixed in 10% formol-saline solution. Gross and fine dissection was carried out to study the incidence of presence of pyramidal lobe, its attachments, length, breadth and association with levator gladulae thyroidae. In the present study, findings were compared with those of Western and Bangladeshi researchers. It was evident that, fifty percent of the thyroids possesses pyramidal lobe. Incidence was more in male (52.1%) than in female (41.7%). It also appeared from the observations that, this lobe was commonly situated on the left side. In the majority of cases (56.66%), base of the pyramidal lobes were attached to the upper border of the isthmus with or without encroachment on the adjacent part of the left lobe. The range of length and breadth of the pyramidal lobe was from 1.8 to 35mm and 1.3 to 19mm respectively. In 73.33% cases, pyramidal lobe was associated with levator glandulae thyroideae and in only 26.66% cases pyramidal lobe was found independently. PMID- 18285724 TI - An anatomical study on interventricular septum of postmortem hearts of adult Bangladeshi people. AB - To study the thickness of interventricular septum (both muscular and membranous parts) of adult Bangladeshi people, fifty postmortem hearts were collected, fixed in 10% formol saline and examined. The thickness of both muscular and membranous parts of the septum was measured. Among fifty hearts, thirty-six were collected from male and fourteen from female cadavers. All hearts were collected from medicolegal cases. The findings were compared between male and female cadavers of our country as well as with the findings of the western countries. The findings suggest that in female, the thickness of both muscular and membranous parts of the septum was less than that of male though there was no significant difference statistically (P>0.05). The findings were compared with the findings of other workers and not so much variation was found. There was no available data regarding the thickness of the membranous part of the septum to compare. PMID- 18285725 TI - Lipid profile changes in second trimester of pregnancy. AB - The present study was carried out in the department of physiology, Mymensingh Medical College, Mymensingh, Bangladesh during the period of July 2006 to June 2007. The aim of the study was to investigate the changes in serum total cholesterol, triacylglycerol, HDL-cholesterol and LDL-cholesterol during the 2nd trimester of normal pregnancy. The serum concentrations of total cholesterol, triacylglycerol, HDL-cholesterol and LDL-cholesterol were measured in 50 cases during the second trimester of pregnancy and in a control group of 100 cases of non pregnant women, which was matched on reproductive age. Data were analyzed by computer with SPSS program using unpaired 't' test. The result showed that the pregnant women had significantly higher concentrations of serum total cholesterol, triacylglycerol and LDL cholesterol (p<0.001). But in case of HDL cholesterol the mean concentration was slightly higher than control group though not statistically significant (p<0.079). Higher concentration of lipids and lipoproteins in second trimester of pregnancy may be a purely physiological response to pregnancy or it may be indicative of pathology in some women. These results warrant a follow up study to investigate whether the hyperlipidaemic response to pregnancy is variable and if so, whether it can predict future hyperlipidaemia in later life. PMID- 18285726 TI - Antigen detection from urine of Kala-azar cases by latex agglutination test. AB - A recently developed Latex agglutination method known as "KATEX" for detecting leishmanial antigen in urine of Kala-azar patients was evaluated on 97 Kala-azar cases and 35 controls in the department of Microbiology, Mymensingh Medical College during the period from March' 2004 to February' 2005. The method yielded sensitivity as 100% and 82.8% in 33 confirmed and 64 ICT positive cases respectively. Since 8.6% controls showed antigen positive results, so specificity of KATEX was calculated as 91.4%. KATEX methods for antigen detection in urine should be used as an early immuno-diagnostic test as it has yielded high sensitivity. But interpretation of a positive test must be made cautiously having correlation with clinical findings as because it becomes false positive in Kala azar free person. Further elucidation of KATEX method including larger population from community giving particular emphasis on its prognostic use was strongly recommended. PMID- 18285727 TI - Comparison of CMV pp65 antigen and IgM antibody detection for early recognition of CMV primary infection and reactivation. AB - Detection of Cytomegalovirus (CMV) pp65 antigen and CMV IgM antibody were compared for the early diagnosis of CMV primary infection or reactivation. Sixty seven immunocompromised patients were studied prospectively for the diagnosis of CMV primary infection or reactivation. CMV IgM antibody was detected in 19 (28.43%) of the 67 immunocompromised patients, whereas pp65 antigen was detected in 20 (29.8%). Among the 19 patients who were positive for both pp65 antigen and CMV IgM antibody, pp65 antigen was detected earlier in 10 of 19 patients. In the remaining 9 patients IgM antibody against CMV was detected simultaneously with the pp65 antigen. The pp65 antigen appeared on an average 16.37 days earlier than that of CMV IgM antibody. PMID- 18285728 TI - Status of informed consent in the original articles published in a medical journal of Bangladesh. AB - Informed consent and other ethical issues are included in every guideline of research ethics. Taking informed consent from the participants is essential in human health research. The health researchers are not addressing adequately the issues of informed consent and other relevant issues. In many of the published articles in our country, there is no mention of informed consent and other relevant ethical issues. In this study, a 5-years retrospective review was performed on the original articles that described research involving human subjects, and published in the "Journal of Bangladesh College of Physicians and Surgeons" in the period of 1999 to 2003. The objectives of this review were: i) to document whether the author has mentioned about informed consent in the published original article; and ii) to document whether the author has addressed the informed consent procedure in appropriate details. Fifty three original articles were reviewed, only in 7(13.2%) articles the author has mentioned that informed consent was taken from the participants. In 19 (63%) articles, where the subjects were minors or incompetent, there was no mention from whom informed consent was taken. Only in 2(3.7%) articles, the authors have mentioned about the approval of Ethical Review Committee, and in 4(7.5%) articles about the funding agency. Other ethical issues were also ignored. The study documents deficiencies in informed consent and relevant ethical issues in the published articles. This finding demands greater attention to ethical standards on the part of investigators and the editors. PMID- 18285729 TI - Medical causes and histological pattern of glomerulonephritis. AB - Glomerulonephritis (GN) is one of the commonest causes of chronic renal failure (CRF) and end stage renal failure (ESRF). The outcome largely depends on the underlying medical cause, clinical findings and histological appearance. 1188 patients were taken in this study. Of these, 980 had primary and 208 secondary GN. Commonest histological types of primary GN were mesangial proliferative glomerulonephritis (31%) and membranous nephropathy (21%). Among patients with secondary GN, predominant cause was lupus nephritis (76%) followed by poststreptococcal GN (18%). Clinically, nephrotic syndrome was present in 54%, hypertension in 39% and renal failure in 28%. But clinical pictures were variable among different histological types of GN. PMID- 18285730 TI - Contraceptive prevalence among adolescent married women in rural Bangladesh. AB - Although contraceptive prevalence rate (CPR) has increased over the last decade it has not yet reached the desired level. This cross sectional study aimed at exploring the current use of the modern methods of contraception and some related issues among the adolescent married girls residing in two rural areas of Faridpur district. A total of 426 girls were interviewed. The mean age of marriage was found to be 15.5+/-1.5 years among the respondents and 57.5% of them had secondary level education. Majority of the women (97.7%) were housewives and 41% of the households had a monthly income of Taka. 2001-4000. It was found that only 41% of the couples were currently practicing a modern method of contraception, oral pill being the most popular method (69.9%). In seventy one percent of the respondents, husband and wife jointly took the decision regarding contraceptive use. The main reasons for use were for spacing purpose (50.3%) and for delaying of first pregnancy (42.2%). Increasing female literacy rate, improving their socio economic condition and providing family planning services at the doorstep could help in improving the situation. PMID- 18285731 TI - Epidemiological study of risk factors related to childhood visceral leishmaniasis. AB - This case control study was carried out to determine certain clinical and epidemiological risk factors related to childhood visceral leishmaniasis (VL). Study was conducted in paediatric wards of Mymensingh Medical College Hospital. Dependable variables are sex, age, housing, use of mosquito net, malnutrition, sleeping habit and seasonality. A total of 60 cases and 60 suitably matched controls were studied between January 2000 to December 2000. The exposure rate of VL is more in boys (70%) than in girls (30%). The frequency rate of VL was higher in children >10 years of age group (66.6%) than in other age groups. Mud house dwelling is significantly associated with the development of VL (81.6%) and carries 28.9 times greater risk of developing VL, than those dwelling in bamboo house (13.3%). The difference of this proportion is statistically significant (p<0.001, OR=28.9). The exposure rate of VL was slightly higher (51.6%) in mosquito net non-users than the mosquito net users. Grade III malnutrition was found significantly associated with (83.3%) VL (p<0.005). The frequency rate of VL was higher in children sleeping on floor than in children sleeping on Chowki or Khat (OR=2.07). Rainy season is significantly associated with VL (60%) and carries 2.4 times greater risk of developing VL than other seasons (p<0.05, OR=2.4). It can be concluded that mud house dwelling, grade III malnutrition, sleeping on floor and seasonality (rainy season) are the important risk factors of VL in children. PMID- 18285732 TI - A giant Condyloma accuminata. AB - Condyloma accuminata is not a rare disease in our country. But such a giant Condyloma accuminata is very uncommon case that we reported here. It is a type of venereal wart that commonly grows in genital region, caused by the HPV (Human papilloma virus). It is considered one of the most common sexually transmitted disease in western countries. The patient presented with cauliflower like fungating growth over the genitalia for 15 years with pain and burning sensation over the lesion for 1 month. Patient had multiple sexual partners for about 20 years. Clinically the patient was diagnosed as Condyloma accuminata. The wart was excised under spinal anesthesia and confirmed by histopathology. PMID- 18285733 TI - Fibrous dysplasia on the skull. AB - A young rickshaw puller admitted in Neurosurgery ward of Mymensingh Medical College Hospital due to ugly looking skull swelling on right side and iatrogenic ulceration on the postero-superior surface of the swelling. He also mentioned that he was not socially well-excepted and facing difficulty in getting married for that ugly lesion and was very unhappy at that time. CT scan was suggestive of fibrous dysplasia of skull bone and no other abnormalities were present on other part of the body. Then complete excision of the mass was done and fibrous dysplasia was confirmed by histopathological study. Two months after surgery he got married and expressed his happiness. In fibrous dysplasia, normal bone is replaced by fibrous connective tissue with varying degrees of osseous metaplasia. It is a benign disorder of bone commonly seen from childhood to the third decade. Virtually any bone in the body can be affected. Regarding investigations, plain radiography in the first line study. CT may be required to assess complex region such as the spine, pelvis, chest and facial skeleton. Histologically, fibrous dysplasia appears as multiple areas of fibrous tissue contained within islands of bone with evidence of both blastic and clastic activity. PMID- 18285734 TI - Cyclic esotropia. AB - Cyclic esotropia is a rare condition in which esotropia alternates with binocular single vision (BSV). A five years old female child presented with the complain of acute onset infrequent squinting of left eye (L/E) for 3 years. Squinting of L/E starts in morning and persists throughout the day. The child becomes completely normal on the following morning without any residual effect. Initially there was 24 hours squinting followed by several months of non-squinting. Gradually interval is lessening. For the last 6 months, she was esotropic for 24 hours and non-esotropic for another 24 hours. She complains of no diplopia during the attack. On examination she is otherwise healthy and hypermetropic by +0.50Dsph on cycloplegic refraction with normal visual acuity (6/6, N5). She was 50 prism diopter esotropic in her left eye on esotropic day both for distant and near fixation and had alternate suppression. Ocular motility is full. Anterior and posterior segment reveals no abnormality. There is BSV with positive stereoscopic vision on Titmus test (40 sec of Arc) on non-esotropic day. This case can be treated surgically either by bilateral medial rectus recession or by recession of medial rectus and resection of lateral rectus of the eye according to the amount of esotropia on the esotropic day. PMID- 18285736 TI - Tubercular splenic abscess. AB - Splenic abscess is a rare clinical condition and yet rarer is a tubercular splenic abscess. Here we report a case of tubercular splenic abscess. A forty years old male patient was admitted in Medicine unit of Mymensingh Medical College Hospital (MMCH) on 09-08-2006 with the complaints of Left upper quadrant abdominal pain and fever for 15 days and Respiratory difficulty for 2 days. Two days after admission he developed generalized abdominal pain and distension. Pain was not associated with vomiting. Patient was transferred to surgical unit for features of peritonitis. Ultrasonogram of whole abdomen revealed moderately enlarged spleen showing 8.8 x 9.7 cm semicystic mass, which may represent an abscess. There was mild free fluid collection in the lower abdomen. X-ray chest P/A view showed bilateral pleural effusion. On laparotomy huge amount of free pus was found in the peritoneal cavity and the spleen was hugely enlarged with a burst abscess cavity in it. Splenectomy and thorough peritoneal toileting was done. Postoperative recovery was uneventful except few stitch infections. Pus culture revealed no growth but histopathology of spleen confirmed Tubercular Splenic Abscess. Patient was given an antitubercular regimen with Rifampicin, Isoniazid, Ethambutol and Pyrazinamide for initial two month which to be followed by Rifampicin and Isoniazid for another ten months. PMID- 18285735 TI - Burst abdomen-A preventable morbidity. AB - A burst abdomen is considered present, when intestine, omentum or other viscera's were seen in the abdominal wound following obstetric surgery. In our country no study found, but observational incidence in the tertiary hospital varies between 0.2-3%. It occurs mostly between the sixth and eight day after operation. Factors relating to the incidence of burst abdomen are suture, closure, incision, coughing, vomiting, distension, obesity, jaundice, malignancy, diabetes mellitus, hypoproteinaemia, anaemia, immuno-compromised patients and wound infection. During the period of February 2001 to February 2006 four cases of burst abdomen were managed in cooperation with team of surgery department. In these cases wound were closed by "May/Mary closure". Abdominal wound dehiscence remains a major cause of morbidity following any laparotomy whether elective or emergency. We should correct the primary risk factors for wound dehiscence. Transverse incisions are generally considered to dehiscence much less than the vertical incision. The suture should have excellent handling and knotting. Its prevention is important to reduce postoperative morbidity, mortality and increased cost of care both in terms of increased hospital stay and treatment of the complication. PMID- 18285737 TI - Fracture of the penis with urethral rupture. AB - We report a rare case of penile fracture with incomplete urethral rupture in a 25 years old male who sustained the injury during sexual intercourse. He presented with a tense haematoma on the ventral aspect of the penile shaft, associated with urethral bleeding. Per urethral catheterization was possible though it was painful. Exploration and repair of the penile fracture and urethra were performed within 16 hrs. The patient made an uneventful recovery with good erectile and voiding function. This case illustrates the value of early surgical repair of the fracture in order to prevent complications. The true incidence of penile fracture is not known even in the Western countries because it is under reported or hidden for social embracement and even it is reported to physicians it remains undiagnosed or mismanaged. Very rarely it is associated with urethral rupture. PMID- 18285738 TI - Stroke in the young: relationship with iron deficiency anemia and thrombocytosis. AB - The recognition and treatment of stroke in children is challenging as there are fundamental developmental differences in children compared to adults. A young boy presented with sudden weakness of right side of body followed by unconsciousness and convulsion that was not preceded by any history of fever or trauma. Neurological examination revealed upper motor neuron lesion in the right extremities. CT scan showed diffuse hypodense area in fronto-parietal regions. Complete blood count and iron profile evaluated iron deficiency anemia with thrombocytosis. Thrombosis resulting from thrombocytosis secondary to iron deficiency anemia was considered as etiological explanation of stroke in this young child. PMID- 18285739 TI - Overlapping of Sturge Weber syndrome and Klippel Trenaunay Weber syndrome. AB - Sturge-Weber Syndrome (SWS) occurs sporadically with a frequency of approximately 1 in 50,000. SWS is a mesodermal phakomatosis. Klippel-Trenaunay Weber syndrome (KTWS) is another very rare phakomatosis. Overlap between SWS & KTWS is very rarely encountered. We report a 19 months old boy with features of both SWS and KTWS. The reported case had seizures, port wine haemangioma of the right side of the body, glaucoma of both eyes, subcortical calcification which were consistent with the Sturge Weber Syndrome; on the other hand he had also hypertrophy of the right side of the including the face and limbs, angiomatous skin naevus, varicosities consistent with the KTWS. PMID- 18285740 TI - Cleidocranial dysplasia. AB - Cleidocranial dysplasia (CCD) is a disorder that affects mostly those bones which derived from endochrondal and intramembranous ossification, such as the cranium and the clavicles. CCD is present at a frequency of one in one million individuals. Over 500 cases have been reported. A 13 years old boy reported in the Paediatric Endocrinology outpatient department, Bangladesh Institute of Research and Rehabilitation on Diabetes, Endocrine and Metabolic Disorders (BIRDEM) on 6th April 2006 with complaints of not growing according to his age, a wide gap over the head since birth and delayed shedding of primary teeth and also eruption of secondary teeth. The boy had elongated facies with midfacial hypoplasia, depressed nasal bridge with hypertelorism. His anterior fontanelle was open about 3/1.5 cm. He had short clavicle and his shoulders could be easily apposed. Radiological findings showed anterior fontanellle and sutures were open and widened, all decidual teeth were intact and all secondary teeth were impacted beneath the primary teeth and hypoplastic clavicle. He was diagnosed as a case of Cleidocranial Dysplasia on the basis of history, clinical examination and investigation. PMID- 18285741 TI - Obturator hernia. AB - An elderly emaciated female patient presented with recurrent lower abdominal pain associated with nausea and vomiting due to obstruction of the small bowel. Although the Howship-Romberg sign and tender mass on digital rectal examination could not ascertained but plain X-ray abdomen shows features of acute intestinal obstruction. After resuscitation laparotomy was done and diagnosed as case of strangulated obturator hernia of Ricters type. Reduction of hernial content and resection of the gangrenous part of small bowel with end-to-end anastomosis done. The hernial defect is repaired by a proleine mesh. Abdomen closed in layers keeping a drain in right iliac fossa and pelvic cavity. The postoperative recovery was uneventful. Patient discharge with advice on 9th postoperative day. PMID- 18285742 TI - Thyrotoxic periodic paralysis. AB - A young man of 30 yrs got himself admitted in Bangabandhu Sheikh Mujib Medical University (BSMMU) Hospital, Dhaka, Bangladesh with the complaints of suddenly developed weakness in his all four limbs. He had features of hyperthyroidism and he gave the history of similar attacks of weakness in his all four limbs in the previous months. His potassium was in the lower part of the normal range and his T4 and T3 were elevated but TSH was markedly low. He was diagnosed as a case of thyrotoxic periodic paralysis on the basis of clinical and biochemical findings. After treatment with carbimazole, propranolol and potassium replacement, patient's condition improved dramatically. PMID- 18285743 TI - Metatropic dysplasia in children. AB - Metatropic dysplasia is a rare genetic condition characterized by progressive dwarfism. Metatropic dysplasia is defined as a short limb skeletal dysplasia characterized by dumbell like configuration of long bones, a narrow but normal length of thorax and occasionally a coccygeal appendage similar to a tail. Children born with this condition show different signs and symptoms throughout the childhood. An eight years old boy was admitted in the Paediatric ward of Bangbandhu Sheikh Mujib Medical University (BSMMU), Dhaka with the complain of swelling in the back for 7 1/2 years. On examination the boy had features suggestive of Metatropic dysplasia. Radiological findings included exaggerated curvature of the spine which was different at the different levels giving rise to "S" shaped deformity, pectus carinatum and flattening of the vertebral bodies which were variable at different levels. From the history, clinical examination and radiological findings the boy was diagnosed as a case of metatropic dysplasia. PMID- 18285744 TI - Tropical pulmonary eosinophilia in a patient undergoing open heart surgery. AB - A 35 years old male living in Jamalpur district of Bangladesh working as a shopkeeper presented with dyspnea, fatigue, occasional fever and cough for two years. He did not give any history of hemoptysis or weight loss. He was smoker and non-alcoholic. He had a soft systolic murmur over pulmonary area and wide fixed splitting of the second heart sound. An Atrial septal defect (ASD) was detected by echocardiography. The patient had high circulating eosinophil count and Complement Fixation Test for filarial antibody revealed positive result. Moreover the patient's response to drug Diethylcarbamazime indicated suspected tropical pulmonary eosinophilia with Atrial Septal Defect. PMID- 18285745 TI - Development of insulin delivery systems. AB - Delivery system of insulin is vital for its acceptance and adherence to therapy for achieving the glycemic targets. Enormous developments have occurred in the delivery system of insulin during the last twenty years and each improvement was aimed at two common goals: patients convenience and better glycemic control. Till to date, the various insulin delivery systems are: syringes/vials, injection aids, jet injectors, transmucosal delivery, transdermal delivery, external insulin infusion pump, implantable insulin pumps, insulin pens and insulin inhalers. Syringe/vial is the oldest and conventional method, still widely used and relatively cheaper. Modern plastic syringes are disposable, light weight with microfine needle for patients convenience and comfort. Oral route could be the most acceptable and viable, if the barriers can be overcome and under extensive trial. Insulin pen device is an important milestone in the delivery system of insulin as it is convenient, discrete, painless, attractive, portable with flexible life style and improved quality of life. More than 80% of European diabetic patients are using insulin pen. Future digital pen will have better memory option, blood glucose monitoring system, insulin dose calculator etc. Insulin infusion pump is a good option for the children, busy patients with flexible lifestyle and those who want to avoid multiple daily injections. Pulmonary route of insulin delivery is a promising, effective, non-invasive and acceptable alternative method. Exubera, the world first insulin inhaler was approved by FDA in 28 January 2006. But due to certain limitations, it has been withdrawn from the market in October 2007. The main concern of inhaled insulin are: long term pulmonary safety issues, cost effectiveness and user friendly device. In future, more acceptable and cost effective insulin inhaler will be introduced. Newer avenues are under extensive trial for better future insulin delivery systems. PMID- 18285746 TI - Even partial remission of proteinuria is associated with better renal outcome in patients with IgA nephropathy. PMID- 18285747 TI - Preventive health care in chronic kidney disease and end-stage renal disease. AB - The complex care that must be provided for patients with renal disease might interfere with provision of basic preventive measures in this population. Preventive health care, including infection screening and prophylaxis, vaccinations, management of blood glucose and lipid levels, and cancer screening, is important, as it might decrease acute morbidity and mortality. This Review highlights useful preventive and health maintenance strategies for patients with chronic kidney disease and those with end-stage renal disease. PMID- 18285748 TI - Management of anemia with intravenous methoxy polyethylene glycol-epoetin beta in patients on dialysis. PMID- 18285749 TI - Botulinum toxin A: a new treatment option for multiple-sclerosis-related bladder overactivity? PMID- 18285750 TI - Investigating vascular myelopathy--when can magnetic resonance angiography replace digital subtraction angiography? PMID- 18285751 TI - Clinical course of neuropathologically confirmed frontal-variant Alzheimer's disease. AB - Background A 66-year-old man presented with a 3-year history of personality changes marked by increasing apathy, social withdrawal and deficits in complex attention, and a 1-year history of progressive memory problems and difficulties in planning and carrying out complex tasks. Investigations Three neuropsychological examinations over 2 years, neurological examination, routine laboratory tests, brain MRI, single-photon emission CT scan, genetic analyses, and neuropathological examination. Diagnosis A clinical diagnosis of frontal variant frontotemporal dementia was superseded by postmortem neuropathological evidence, which established a diagnosis of frontal-variant Alzheimer's disease. Management The patient and his spouse were referred for counseling, and the patient was referred for follow-up examinations. PMID- 18285752 TI - Active surveillance for low-risk prostate cancer: selection of patients and predictors of progression. AB - The natural history of prostate cancer is often of long duration, and the disease is incompletely understood. Whether all men with prostate cancer require immediate treatment or whether men with tumors of low malignant potential are being overtreated with potentially harmful therapies is a subject of much debate. Results from a randomized trial that compared watchful waiting and active therapy showed all-cause and disease-specific survival advantages with radical therapy, but the study group was mixed in terms of disease risk; the optimum treatment strategy for men with low risk features remains unclear. Multiple centers are gaining experience with active surveillance and delayed intervention with curative intent for men with prostate tumors of potentially low clinical risk. This Review describes the background studies behind the rationale for active surveillance, thoughts on selection criteria for candidates and some early reported outcomes for active surveillance cohorts. The psychosocial impact of active surveillance on patients is discussed as well as contemporary methods for disease monitoring. PMID- 18285753 TI - Predictors of biochemical progression-free survival in men who receive radiation therapy after prostatectomy. PMID- 18285754 TI - Does antimicrobial prophylaxis affect recurrent urinary tract infections and bacterial resistance in children? PMID- 18285755 TI - Intermediate phenotypes in schizophrenia genetics redux: is it a no brainer? PMID- 18285757 TI - Oral administration of the antiobesity drugs, sibutramine and rimonabant, increases acetylcholine efflux selectively in the medial prefrontal cortex of the rat. PMID- 18285758 TI - Association of polymorphisms in the angiotensin-converting enzyme gene with syndromal panic attacks. PMID- 18285759 TI - Do autism-related personality traits explain higher paternal age in autism? PMID- 18285760 TI - Bevacizumab in combination with irinotecan for patients with recurrent glioblastoma multiforme. PMID- 18285761 TI - A comparison of chemotherapy plus surgery with surgery alone for localized esophageal cancer. PMID- 18285762 TI - Methotrexate-associated mantle-cell lymphoma in an elderly man with myasthenia gravis. AB - BACKGROUND: A 75-year-old man on methotrexate immunosuppression for myasthenia gravis presented with a 2-month history of lymphocytosis and bilateral inguinal adenopathy. There were no constitutional symptoms of fever, night sweats, or weight loss. INVESTIGATIONS: Physical examination, blood tests, flow cytometry, fluorescent in situ hybridization, immunoglobulin gene sequencing, viral load quantification by real-time polymerase chain reaction, excisional lymph-node biopsy, bone-marrow biopsy, tumor morphology and immunohistochemistry, sequential CT and PET scans. DIAGNOSIS: Methotrexate-associated mantle-cell lymphoma. MANAGEMENT: Cessation of methotrexate, anthracycline-based combination chemo immunotherapy, and maintenance rituximab. PMID- 18285763 TI - Are NSAIDs and selective cyclo-oxygenase 2 inhibitors associated with increased risk of myocardial infarction? AB - Scott et al. conducted a systematic review to evaluate the risk of myocardial infarction (MI) associated with cyclo-oxygenase-2 (COX2) inhibitors and traditional NSAIDs. The review found a small increased risk of MI associated with COX2 inhibitors, particularly rofecoxib. Although a fixed-effects model of 14 case-control studies suggested a slightly increased risk of MI with NSAID use, a random-effects model of the same data and analysis of 6 cohort studies found no such link. An increased risk of MI was found in four RCTs of NSAID use in colonic adenoma. In an analysis of 14 RCTs that compared COX2 inhibitors with traditional NSAIDs in patients with arthritis, the odds ratio for MI with COX2 inhibitors was 1.6 (95% CI 1.1-2.4), although most of this risk was accounted for by rofecoxib. COX2 inhibitors were, however, associated with a reduced risk of serious gastrointestinal events. An analysis of previous systematic reviews showed increased risks of MI associated with rofecoxib and celecoxib. PMID- 18285764 TI - Technology Insight: hematopoietic stem cell transplantation for systemic rheumatic disease. AB - Hematopoietic stem cells (HSCs) have the capacity for self-renewal and the potential to differentiate into all types of hematopoietic and immune system cells. These features have been successfully used to treat a multitude of hematologic malignancies and nonmalignant diseases such as aplastic anemia, hemoglobinopathies, inborn errors of metabolism and congenital immunodeficiency states. The application of HSC transplantation has been expanded over the past decade to include immune-mediated diseases such as multiple sclerosis, treatment refractory rheumatoid arthritis, systemic lupus erythematosus and systemic sclerosis. Transplantation of HSCs for the treatment of autoimmune diseases aims to fundamentally correct the dysregulated immune system, which could result in sustained clinical remission or potential cure. The use of this approach is currently restricted to clinical research, as there is no standard conditioning regimen to attain these aims in autoimmune diseases. HSC transplantation is associated with inherent morbidity and mortality, both treatment-related and disease-related, and selecting the correct group of patients with the best risk:benefit ratio is a challenging task. PMID- 18285766 TI - Politicizing science no more. PMID- 18285767 TI - Immunological outreach through positive selection. PMID- 18285765 TI - Mechanisms of disease: antiphospholipid antibodies-from clinical association to pathologic mechanism. AB - The discovery that antiphospholipid antibodies recognize plasma proteins that bind to phospholipids rather than recognizing phospholipids themselves has been a major advance in research into antiphospholipid syndrome (APS). It is now established that beta2-glycoprotein I (beta2 GPI) is the most important antigen for antiphospholipid antibodies. However, the possible pathologic mechanism is still much debated. This is mainly because not all patients with anti-beta2 GPI antibodies show clinical symptoms that are related to APS. Several reports indicate that anti-beta2 GPI antibodies with lupus anticoagulant (LA) activity are clinically of much importance. Most patients with LA caused by anti-beta2 GPI antibodies suffer from thrombosis as a result of recognition of the first domain of beta2 GPI by these antibodies. In the search for a pathologic mechanism that might explain the high occurrence of thrombosis in patients with anti-domain I antibodies (LA-causing anti-beta2 GPI antibodies), it was found that these antibodies show increased resistance to the anticoagulant activity of annexin A5. We have shown that the same population of antibodies also displays increased resistance to activated protein C. Owing to the diversity of clinical symptoms related to APS, it is likely that other pathologic mechanisms also contribute to the occurrence of APS-related symptoms. PMID- 18285768 TI - Basophils: in the spotlight at last. PMID- 18285769 TI - HIV's gut feeling. PMID- 18285770 TI - Itching to end NF-kappaB. PMID- 18285771 TI - How T cells 'find' the right dendritic cell. PMID- 18285772 TI - Finally, MALT1 is a protease! PMID- 18285773 TI - Innocents abroad: regulating where naive T cells go. PMID- 18285779 TI - Pharmacoecology: a new name for an old science. PMID- 18285775 TI - The Foxp3+ regulatory T cell: a jack of all trades, master of regulation. AB - The function of regulatory T cells (T(reg) cells) has been attributed to a growing number of diverse pathways, molecules and processes. Seemingly contradictory conclusions regarding the mechanisms underlying T(reg) cell suppressive activity have revitalized skeptics in the field who challenge the core validity of the idea of T(reg) cells as central immune regulators. However, we note that a consensus may be emerging from the data: that multiple T(reg) cell functions act either directly or indirectly at the site of antigen presentation to create a regulatory milieu that promotes bystander suppression and infectious tolerance. Thus, the versatility and adaptability of the Foxp3+ T(reg) cells may in fact be the best argument that these cells are 'multitalented masters of immune regulation'. PMID- 18285783 TI - Special cells, special considerations: the challenges of bringing embryonic stem cells from the laboratory to the clinic. PMID- 18285784 TI - Why the United States still needs improved dietary supplement regulation and oversight. AB - It has been 3 years since the American Society for Clinical Pharmacology and Therapeutics (ASCPT) issued a position statement regarding dietary supplement safety and regulation. I was the Chair of the ASCPT task force charged with issuing the statement. At the time, after careful review of available data, the other members and I concluded that dietary supplement legislation in the United States was lacking and that enhanced oversight was essential to increase the safety of these products for the American consumer. PMID- 18285785 TI - Should herbal medicines be regulated as drugs? PMID- 18285786 TI - Drug-drug interaction programs in clinical practice. AB - There are multiple publications and electronic programs available to aid clinicians in the management of drug-drug interactions; however, clinicians should understand the advantages and limitations of these programs before using them in clinical practice. Because time constraints can make evaluation of the primary literature challenging, clinicians and health-care providers frequently rely on drug-interaction computer programs that are integrated in the prescribing process to flag drug interactions. PMID- 18285788 TI - Abstracts of the Annual Meeting of the American Society for Clinical Pharmacology and Therapeutics. Orlando, Florida, USA. April 2-5, 2008. PMID- 18285800 TI - The pathophysiological basis of dystonias. AB - Dystonias comprise a group of movement disorders that are characterized by involuntary movements and postures. Insight into the nature of neuronal dysfunction has been provided by the identification of genes responsible for primary dystonias, the characterization of animal models and functional evaluations and in vivo brain imaging of patients with dystonia. The data suggest that alterations in neuronal development and communication within the brain create a susceptible substratum for dystonia. Although there is no overt neurodegeneration in most forms of dystonia, there are functional and microstructural brain alterations. Dystonia offers a window into the mechanisms whereby subtle changes in neuronal function, particularly in sensorimotor circuits that are associated with motor learning and memory, can corrupt normal coordination and lead to a disabling motor disorder. PMID- 18285801 TI - Functional neuroimaging of the vegetative state. AB - A number of recent studies have demonstrated a role for state-of-the-art neuroimaging methods in the assessment of patients in the vegetative state and other so-called 'disorders of consciousness'. In several cases, functional MRI has been used to show that aspects of speech perception, emotional processing, language comprehension and even conscious awareness might be retained in some patients who behaviourally meet all of the criteria that define the vegetative state. This work has profound implications for clinical care, diagnosis, prognosis and medical-legal decision making (relating to the prolongation, or otherwise, of life after severe brain injury), as well as for more basic scientific questions about the nature of consciousness and the neural representation of our own thoughts and intentions. PMID- 18285803 TI - Regulation of DNA repair throughout the cell cycle. AB - The repair of DNA lesions that occur endogenously or in response to diverse genotoxic stresses is indispensable for genome integrity. DNA lesions activate checkpoint pathways that regulate specific DNA-repair mechanisms in the different phases of the cell cycle. Checkpoint-arrested cells resume cell-cycle progression once damage has been repaired, whereas cells with unrepairable DNA lesions undergo permanent cell-cycle arrest or apoptosis. Recent studies have provided insights into the mechanisms that contribute to DNA repair in specific cell-cycle phases and have highlighted the mechanisms that ensure cell-cycle progression or arrest in normal and cancerous cells. PMID- 18285802 TI - The role of oxygen availability in embryonic development and stem cell function. AB - Low levels of oxygen (O2) occur naturally in developing embryos. Cells respond to their hypoxic microenvironment by stimulating several hypoxia-inducible factors (and other molecules that mediate O2 homeostasis), which then coordinate the development of the blood, vasculature, placenta, nervous system and other organs. Furthermore, embryonic stem and progenitor cells frequently occupy hypoxic 'niches' and low O2 regulates their differentiation. Recent work has revealed an important link between factors that are involved in regulating stem and progenitor cell behaviour and hypoxia-inducible factors, which provides a molecular framework for the hypoxic control of differentiation and cell fate. These findings have important implications for the development of therapies for tissue regeneration and disease. PMID- 18285804 TI - Dietary blueberry supplementation affects growth but not vascularization of neural transplants. AB - Transplantation of neural tissue has been attempted as a treatment method for neurodegenerative disorders. Grafted neurons survive to a lesser extent into middle-aged or aged hosts, and survival rates of <10% of grafted neurons is common. Antioxidant diets, such as blueberry, can exert powerful effects on developing neurons and blood vessels in vitro, but studies are lacking that examine the effects of these diets on transplanted tissues. In this study, we examined the effects of a blueberry diet on survival, growth, and vascularization of fetal hippocampal tissue to the anterior chamber of the eye of young or middle aged female rats. Previous work from our group showed significant increase in neuronal survival and development with blueberry diet in grafts. However, the effects of antioxidant diet on vascular development in grafts have not been explored previously. The age of the host affected individual vessel morphology in that aged hosts contained grafts with thick, undeveloped walls, and wider lumen. The blood-brain barrier also appeared to be affected by the age of the host. The blueberry diet did not affect vessel morphology or density of vessel-associated protein markers but gave rise to significantly increased growth capacity, cytoarchitecture, and the final size of hippocampal grafts. PMID- 18285806 TI - Cross-validation of bioelectrical impedance analysis for the assessment of body composition in a representative sample of 6- to 13-year-old children. AB - BACKGROUND/OBJECTIVES: (1) To cross-validate tetra- (4-BIA) and octopolar (8-BIA) bioelectrical impedance analysis vs dual-energy X-ray absorptiometry (DXA) for the assessment of total and appendicular body composition and (2) to evaluate the accuracy of external 4-BIA algorithms for the prediction of total body composition, in a representative sample of Swiss children. SUBJECTS/METHODS: A representative sample of 333 Swiss children aged 6-13 years from the Kinder Sportstudie (KISS) (ISRCTN15360785). Whole-body fat-free mass (FFM) and appendicular lean tissue mass were measured with DXA. Body resistance (R) was measured at 50 kHz with 4-BIA and segmental body resistance at 5, 50, 250 and 500 kHz with 8-BIA. The resistance index (RI) was calculated as height(2)/R. Selection of predictors (gender, age, weight, RI4 and RI8) for BIA algorithms was performed using bootstrapped stepwise linear regression on 1000 samples. We calculated 95% confidence intervals (CI) of regression coefficients and measures of model fit using bootstrap analysis. Limits of agreement were used as measures of interchangeability of BIA with DXA. RESULTS: 8-BIA was more accurate than 4 BIA for the assessment of FFM (root mean square error (RMSE)=0.90 (95% CI 0.82 0.98) vs 1.12 kg (1.01-1.24); limits of agreement 1.80 to -1.80 kg vs 2.24 to 2.24 kg). 8-BIA also gave accurate estimates of appendicular body composition, with RMSE < or = 0.10 kg for arms and < or = 0.24 kg for legs. All external 4-BIA algorithms performed poorly with substantial negative proportional bias (r> or = 0.48, P<0.001). CONCLUSIONS: In a representative sample of young Swiss children (1) 8-BIA was superior to 4-BIA for the prediction of FFM, (2) external 4-BIA algorithms gave biased predictions of FFM and (3) 8-BIA was an accurate predictor of segmental body composition. PMID- 18285805 TI - Meat consumption after disaggregation of meat dishes in a cohort of British adults in 1989 and 1999 in relation to diet quality. AB - BACKGROUND/OBJECTIVES: The aim of the study was to quantify more precisely the meat intake of a cohort of adults in the UK by disaggregating composite meat dishes. SUBJECTS/METHODS: Subjects were members of the Medical Research Council National Survey of Health and Development, 1946 birth cohort. Five-day diaries were collected from 2256 men and women in 1989 and 1772 men and women in 1999. From the details provided, composite meat dishes were broken down into their constituent parts and the meat fraction was added to meat portions only. Meat intake was classified as red meat, processed meat and poultry. RESULTS: Meat consumption without disaggregation of meat dishes resulted in a mean overestimation of 50% in men and 33% in women. Red meat consumption fell between 1989 and 1999 from 51.7 to 41.5 g per day in men and 35.7 to 30.1 g per day in women. Poultry consumption rose from 21.6 to 32.2 g per day in men and 18.2 to 29.4 g per day in women. Re-calculating red meat intakes resulted in the percentage of subjects in 1999 consuming more than the recommendation of the World Cancer Research Fund falling from 30 to 12%. Increasing consumption of red and processed meat was associated with increased intakes of energy, fat, haem iron, zinc and vitamin B(12), and lower intake of fibre. Increased sodium intake was associated with increased consumption of processed meat. CONCLUSIONS: Disaggregation of meat dishes provided a more precise estimate of meat consumption. The quantity of red or processed meat in the diet was reflected in the nutrient content of the entire diet. PMID- 18285807 TI - Breast feeding in infancy and arterial endothelial function later in life. The Cardiovascular Risk in Young Finns Study. AB - BACKGROUND/OBJECTIVES: Breast feeding in infancy may be associated with reduced cardiovascular morbidity in adulthood. We examined the association between breast feeding in infancy and arterial function and structure in adulthood in a population-based cohort of Finnish adults. SUBJECTS/METHODS: Noninvasive ultrasound was used to measure brachial artery flow-mediated dilatation (FMD), carotid artery intima-media thickness (IMT) and carotid artery compliance (CAC) in 1667 young adults participating in the Cardiovascular Risk in Young Finns Study with data on early nutrition. RESULTS: Maximal FMD was higher in breast-fed men compared to formula-fed men (7.2+/-4.0 vs 5.9+/-3.4%, P=0.029) while no differences were seen between breast-fed and formula-fed women (8.9+/-4.5 vs 8.8+/-5.0%, P=0.84). In men, the multivariable correlates of FMD included the group variable for breast feeding (P=0.014), birth weight (P=0.043), waist circumference (P<0.001) and baseline brachial artery diameter (P<0.001). In women, the multivariable correlates of FMD were birth weight (P=0.02), waist circumference (P<0.001) and brachial artery baseline diameter (P<0.001). Breast feeding was not significantly associated with IMT or CAC in multivariable models. CONCLUSIONS: Adult men who have been breast fed have better brachial endothelial function compared to men who have been formula fed. PMID- 18285808 TI - Testing nutrient profile models in relation to energy density and energy cost. AB - BACKGROUND: Nutrient profiling of foods is defined as the science of classifying foods based on their nutrient content. Food rankings generated by nutrient profile models need to be tested against objective reality as opposed to public opinion. OBJECTIVE: To test the performance of selected nutrient profile models in relation to the foods' energy density (kcal g(-1)) and energy cost (Dollar per 1000 kcal). SUBJECTS/METHODS: Analyses were based on 378 component foods of a food frequency instrument. The models tested were the French nutrient adequacy models NAS23 and NAS16 and nutrient density models NDS23 and NDS16; and a family of nutrient-rich models (NR(n), where n=5-7; 10-12, and 15). Also tested were LIM scores and a modified British Food Standards Agency model WXYfm. Profiles were calculated based on 100 g, 100 kcal and on Reference Amounts Customarily Consumed. Food rankings generated by different models were correlated with each other and with the foods' energy density and energy cost. RESULTS: Nutrient profile models based on protein, fiber, vitamins and minerals showed an inverse correlation with energy density that diminished as more micronutrients were introduced into the model. Models based on fat, sugar and sodium were highly correlated with energy density. Foods classified as healthier were generally associated with higher energy costs. CONCLUSIONS: Not all models accurately reflected the foods' content of nutrients known to be beneficial to health. High correlations with energy density meant that some models classified foods based on their energy density as opposed to nutrient content. PMID- 18285809 TI - Vitamin D insufficiency is common in Indian mothers but is not associated with gestational diabetes or variation in newborn size. AB - BACKGROUND/OBJECTIVES: Vitamin D is required for bone growth and normal insulin secretion. Maternal hypovitaminosis D may impair fetal growth and increase the risk of gestational diabetes. We have related maternal vitamin D status in pregnancy to maternal and newborn glucose and insulin concentrations, and newborn size, in a South Indian population. SUBJECTS/METHODS: Serum 25 hydroxy vitamin D (25(OH)D) concentrations, glucose tolerance, and plasma insulin, proinsulin and 32-33 split proinsulin concentrations were measured at 30 weeks gestation in 559 women who delivered at the Holdsworth Memorial Hospital, Mysore. The babies' anthropometry and cord plasma glucose, insulin and insulin precursor concentrations were measured. RESULTS: In total 66% of women had hypovitaminosis D (25(OH)D concentrations <50 nmol l(-1)) and 31% were below 28 nmol l(-1). There was seasonal variation in 25(OH)D concentrations (P<0.0001). There was no association between maternal 25(OH)D and gestational diabetes (incidence 7% in women with and without hypovitaminosis D). Maternal 25(OH)D concentrations were unrelated to newborn anthropometry or cord plasma variables. In mothers with hypovitaminosis D, higher 25(OH)D concentrations were associated with lower 30 min glucose concentrations (P=0.03) and higher fasting proinsulin concentrations (P=0.04). CONCLUSIONS: Hypovitaminosis D at 30 weeks gestation is common in Mysore mothers. It is not associated with an increased risk of gestational diabetes, impaired fetal growth or altered neonatal cord plasma insulin secretory profile. PMID- 18285810 TI - Resting energy expenditure can be assessed by dual-energy X-ray absorptiometry in women regardless of age and fitness. AB - OBJECTIVE: To evaluate the possibility that measurement of the magnitude and distribution of fundamental somatic heat-producing units using dual-energy X-ray absorptiometry (DXA) can be used to estimate resting energy expenditure (REE) in both young and elderly women with different aerobic fitness levels. SUBJECTS AND METHODS: Peak oxygen uptake (VO(2) peak) and REE(m) were directly measured in 116 young (age: 22.3+/-2.1 years) and 72 elderly (63.3+/-6.4 years) women. The subjects were divided into four groups according to categories of age and VO(2) peak; young: high fitness (YH, n=58); low fitness (YL, n=58); elderly: high fitness (EH, n=37) and low fitness (EL, n=35). Using DXA, systemic and regional body compositions were measured, and REE(e) was estimated from the sum of tissue organ weights multiplied by corresponding metabolic rate. RESULTS: Although there were remarkable differences in systemic and regional body compositions, no significant differences were observed between REE(m) and REE(e) in the four groups. REE(e) significantly correlated with REE(m) in elderly as well as young women; the slopes and intercepts of the two regression lines were statistically not different between the elderly and young groups (elderly: y=0.60x + 472, r=0.667; young: y=0.78x + 250, r=0.798; P<0.001, respectively). A Bland-Altman analysis did not indicate bias in calculation of REE for all the subjects. CONCLUSION: These results suggest that REE can be estimated from tissue organ components in women regardless of age and aerobic fitness. PMID- 18285811 TI - How multilocus genotypic pattern helps to understand the history of selfing populations: a case study in Medicago truncatula. AB - The occurrence of populations exhibiting high genetic diversity in predominantly selfing species remains a puzzling question, since under regular selfing genetic diversity is expected to be depleted at a faster rate than under outcrossing. Fine-scale population genetics approaches may help to answer this question. Here we study a natural population of the legume Medicago truncatula in which both the fine-scale spatial structure and the selfing rate are characterized using three different methods. Selfing rate estimates were very high ( approximately 99%) irrespective of the method used. A clear pattern of isolation by distance reflecting small seed dispersal distances was detected. Combining genotypic data over loci, we could define 34 multilocus genotypes. Among those, six highly inbred genotypes (lines) represented more than 75% of the individuals studied and harboured all the allelic variation present in the population. We also detected a large set of multilocus genotypes resembling recombinant inbred lines between the most frequent lines occurring in the population. This finding illustrates the importance of rare recombination in redistributing available allelic diversity into new genotypic combinations. This study shows how multilocus and fine-scale spatial analyses may help to understand the population history of self fertilizing species, especially to make inferences about the relative role of foundation/migration and recombination events in such populations. PMID- 18285812 TI - Mitochondrial and nuclear DNA variability in the living fossil Triops cancriformis (Bosc, 1801) (Crustacea, Branchiopoda, Notostraca). AB - The living fossil Triops cancriformis comprises bisexual (either gonochoric or hermaphroditic) and unisexual populations. Genetic surveys have recently revealed a general trend of low differentiation of 12S and 16S mitochondrial genes. We, therefore, surveyed further mitochondrial (COI gene and control region) and nuclear markers (dinucleotide microsatellites) to assess the genetic variability and to establish any relationship with the different reproductive modes found in European populations. The mitochondrial analyses confirmed the pattern of low variability. Hence, the low mitochondrial genetic variability appears as a common feature of the genus Triops. The microsatellite analysis found that Italian populations are monomorphic or exhibit little polymorphism, while other European samples display a higher degree of polymorphism and private alleles. Spanish, Austrian and Italian populations show patterns of Hardy-Weinberg disequilibrium that could be explained by the mode of reproduction, or by a higher frequency of null alleles in these populations. The low diversity and differentiation among Italian populations lead us to question the Monopolization Hypothesis. One microsatellite locus appears to be sex-linked, with heterozygotes detected only in males and hermaphrodites. PMID- 18285813 TI - One-sided evolution or two? A reply to Ennos. PMID- 18285814 TI - Genetics of variation in adiponectin in pedigreed baboons: evidence for pleiotropic effects on adipocyte volume and serum adiponectin. AB - To detect and localize the effects of genes influencing variation in adiponectin mRNA and protein levels, we conducted statistical genetic analyses of circulating concentrations of adiponectin and adiponectin (ADIPOQ) mRNA expression in omental adipose tissue in adult, pedigreed baboons (Papio anubis). An omental adipose tissue biopsy and blood sample were collected from 427 baboons from the colony at the Southwest Foundation for Biomedical Research, San Antonio, TX. Total RNA was isolated from adipose tissue and adiponectin mRNA levels were assayed by real time, quantitative reverse transcriptase-PCR. Adiponectin, insulin, glucose, cholesterol, high-density lipoproteins and triglycerides were measured in fasting serum. Quantitative genetic analyses were conducted for adiponectin mRNA and serum protein using a maximum likelihood-based variance decomposition approach. A genome-wide linkage analysis was conducted using adiponectin mRNA and protein levels as phenotypes. Significant heritability was estimated for ADIPOQ mRNA levels (h2=0.19+/-0.07, P=0.01) and protein levels (h2=0.28+/-0.14, P=0.003). Genetic correlations were found between adiponectin protein and body weight (rho(G)=-0.51, P=0.03), cell volume (rho(G)=-0.73, P=0.04), serum triglycerides (rho(G)=-0.67, P=0.03), and between adiponectin mRNA and glucose (rho(G)=0.93, P<0.01). A logarithm of odds score of 2.9 was found for ADIPOQ mRNA levels on baboon chromosome 4p, which is orthologous to human 6p21. There is a significant genetic component affecting variation in the analyzed traits, and common genes may be influencing adiponectin expression, adipocyte volume, body weight and circulating triglycerides. The region on 6p21 has been linked to diabetes-related phenotypes in human studies. PMID- 18285815 TI - Is lansoprazole effective for the initial management of young patients with dyspepsia? PMID- 18285816 TI - Benefits and risks of a fish diet--should we be eating more or less? PMID- 18285817 TI - Endoscopic variceal ligation--is there any benefit to placing more than six bands? PMID- 18285818 TI - Using heparin therapy to reverse protein-losing enteropathy in a patient with CDG Ib. AB - BACKGROUND: A 22-year-old female presented with edema, diarrhea, hypoalbuminemia and pancytopenia. She had previously been diagnosed with congenital disorder of glycosylation type Ib, and had a history of congenital hepatic fibrosis, portal hypertension and esophageal varices. In the past she had refused mannose therapy because of associated diarrhea and abdominal pain. INVESTIGATIONS: Laboratory examinations, abdominal ultrasonography, bacterial and viral cultures of blood, urine and stools, double-balloon enteroscopy and fecal excretion test using 51Cr labeled albumin. DIAGNOSIS: Protein-losing enteropathy. MANAGEMENT: Infusion of albumin followed by intravenous and subcutaneous therapy with unfractionated heparin. PMID- 18285819 TI - Quality control of DNA break metabolism: in the 'end', it's a good thing. AB - DNA ends pose specific problems in the control of genetic information quality. Ends of broken DNA need to be rejoined to avoid genome rearrangements, whereas natural DNA ends of linear chromosomes, telomeres, need to be stable and hidden from the DNA damage response. Efficient DNA end metabolism, either at induced DNA breaks or telomeres, does not result from the machine-like precision of molecular reactions, but rather from messier, more stochastic processes. The necessary molecular interactions are dynamically unstable, with constructive and destructive processes occurring in competition. In the end, quality control comes from the constant building up and tearing down of inappropriate, but also appropriate reaction steps in combination with factors that only slightly shift the equilibrium to eventually favour appropriate events. Thus, paradoxically, enzymes antagonizing DNA end metabolism help to ensure that genome maintenance becomes a robust process. PMID- 18285821 TI - Revised spectrum of mutations in sarcoglycanopathies. AB - To define the spectrum of mutations in alpha-, beta-, gamma-, and delta sarcoglycan (SG) genes, we analyzed these genes in 69 probands with clinical and biological criteria compatible with the diagnosis of autosomal recessive limb girdle muscular dystrophy. For 48 patients, muscle biopsies were available and multiplex western blot analysis of muscle proteins showed significant abnormalities of alpha- and gamma-SG. Our diagnostic strategy includes multiplex western blot, sequencing of SG genes, multiplex quantitative-fluorescent PCR and RT-PCR analyses. Mutations were detected in 57 patients and homozygous or compound heterozygous mutations were identified in 75% (36/48) of the patients with abnormal western blot, and in 52% (11/21) of the patients without muscle biopsy. Involvement of alpha-SG was demonstrated in 55.3% of cases (26/47), whereas gamma- and beta-SG were implicated in 25.5% (12/47) and in 17% (8/47) of cases, respectively. Interestingly, we identified 25 novel mutations, and a significant proportion of these mutations correspond to deletions (identified in 14 patients) of complete exon(s) of alpha- or gamma-SG genes, and partial duplications (identified in 5 patients) of exon 1 of beta-SG gene. This study highlights the high frequency of exonic deletions of alpha- and gamma-SG genes, as well as the presence of a hotspot of duplications affecting exon 1 of the beta SG gene. In addition, protein analysis by multiplex western blot in combination with mutation screening and genotyping results allowed to propose a comprehensive and efficient diagnostic strategy and strongly suggested the implication of additional genes, yet to be identified, in sarcoglycanopathy-like disorders. PMID- 18285822 TI - XPC polymorphisms play a role in tissue-specific carcinogenesis: a meta-analysis. AB - XPC participates in the initial recognition of DNA damage during the DNA nucleotide excision repair process in global genomic repair. Polymorphisms in XPC gene have been analyzed in case-control studies to assess the cancer risk attributed to these variants, but results are conflicting. To clarify the impact of XPC polymorphisms in cancer risk, we performed a meta-analysis that included 33 published case-control studies. Polymorphisms analyzed were Lys939Gln and Ala499Val. The overall summary odds ratio (OR) for the associations of the 939Gln/Gln genotype with risk of cancer was 1.01 (95% confidence interval (95% CI): 0.94-1.09), but there were statistically significant associations for lung cancer, observed for the recessive genetic model (Lys/Lys+Lys/Gln vs Gln/Gln), (OR 1.30; 95% CI: 1.113-1.53), whereas for breast cancer a reduced but nonsignificant risk was observed for the same model (OR 0.87; 95% CI: 0.74-1.01). The results for Ala499Val showed a significant overall increase in cancer risk (OR 1.15; 95% CI: 1.02-1.31), and for bladder cancer in both the simple genetic model (Ala/Ala vs Val/Val) (OR 1.30; 95% CI: 1.04-1.61) and the recessive genetic model (Ala/Ala+Ala/Val vs Val/Val) (OR 1.32; 95% CI: 1.06-1.63). Our meta analysis supports that polymorphisms in XPC may represent low-penetrance susceptibility gene variants for breast, bladder, head and neck, and lung cancer. XPC is a good candidate for large-scale epidemiological case-control studies that may lead to improvement in the management of highly prevalent cancers. PMID- 18285823 TI - Hereditary hemorrhagic telangiectasia: evidence for regional founder effects of ACVRL1 mutations in French and Italian patients. AB - Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disease characterized by widespread arteriovenous malformations and caused by mutations in two major genes: ENG and ACVRL1. Two decades ago, a French epidemiological study pointed out that its prevalence was higher than previously thought and that its distribution varied greatly from one area to another, one of the highest concentrations of patients being found in the Haut-Jura mountains. Although germline mutations are usually family specific, some of them have been reported in unrelated patients, especially for ACVRL1. We performed haplotype analysis of 116 French and Italian patients carrying 13 ACVRL1 different mutations. For five of these mutations, we estimated the age of the most recent common ancestors (MRCAs) using the ESTIAGE program. Most mutations were related to both recurrent mutational events and founder effects with age estimates ranging from 100 to 550 years. The c.1112dupG mutation, which is likely to be responsible for the very high concentration of HHT patients found in the former epidemiological study, probably occurred in one inhabitant of the Haut-Jura Mountains more than three centuries ago. The p.Arg374Gln mutation occurred independently in at least two distinct geographical areas, including the area with the second highest prevalence in the epidemiological study and where the MRCA is rather recent (about 100 years ago). Partially shared haplotypes between French and Italian patients were found for three mutations. This suggests a common origin and a possible diffusion of these mutations from Italy to France. PMID- 18285824 TI - A genome-wide linkage scan for low spinal bone mineral density in a single extended family confirms linkage to 1p36.3. AB - Osteoporotic fractures are an increasing cause of mortality and morbidity in ageing populations. A major risk determinant for these fractures is bone mineral density (BMD). Variation on BMD is thought, on the basis of twin and family studies, to be subject to a large amount of genetic variation and it has been hypothesised that this may be due to the influence of multiple genes. However, in families showing segregation of low or high BMD, single major genes have been shown to play a crucial role. We performed a genome-wide screen using 380 microsatellite markers in a single extended family (n=34) in which early-onset low spinal areal BMD segregates in an autosomal dominant-like fashion. A two point linkage analysis was performed, revealing a maximum LOD score of 3.07 on 1p36.3 (D1S468), confirming results of previous linkage studies of BMD, while no other suggestive linkage peaks (LOD>2.2) were detected elsewhere in the genome. Microsatellite markers were subsequently genotyped for a +/-6.9 Mb region surrounding D1S468. This revealed critical recombination events restricting the candidate region to 1.2 Mb and 19 genes. Sequencing analysis of the coding region of candidate genes WDR8 and EGFL3 revealed no mutations or disease-associated polymorphisms. Our results provide some evidence supporting the hypothesis that there are genetic determinants for spinal BMD on 1p36.3. Although no specific disease causing mutation has yet been found, the delineation of a relatively small candidate region in a single extended family opens perspectives to identify a major gene for spinal BMD. PMID- 18285825 TI - A mutational analysis of the SLC26A4 gene in Spanish hearing-impaired families provides new insights into the genetic causes of Pendred syndrome and DFNB4 hearing loss. AB - Pendred syndrome (PS) and DFNB4, a non-syndromic sensorineural hearing loss with enlargement of the vestibular aqueduct (EVA), are caused by mutations in the SLC26A4 gene. Both disorders are recessive, and yet only one mutated SLC26A4 allele, or no mutations, are identified in many cases. Here we present the genetic characterization of 105 Spanish patients from 47 families with PS or non syndromic EVA and 20 families with recessive non-syndromic hearing loss, which segregated with the DFNB4 locus. In this cohort, two causative SLC26A4 mutations could be characterized in 18 families (27%), whereas a single mutated allele was found in a patient with unilateral hearing loss and EVA in the same ear. In all, 24 different causative mutations were identified, including eight novel mutations. The novel p.Q514K variant was the most prevalent mutation in SLC26A4, accounting for 17% (6/36) of the mutated alleles identified in this study, deriving from a founder effect. We also characterized a novel multiexon 14 kb deletion spanning from intron 3 to intron 6 (g.8091T_22145Cdel). This study also revealed the first case of a de novo recessive mutation p.Q413P causing PS that arose in the proband's paternal allele, the maternal one carrying the p.L445W. The relevance of our results for genetic diagnosis of PS and non-syndromic EVA hearing loss is discussed. PMID- 18285820 TI - DNA-damage repair; the good, the bad, and the ugly. AB - Organisms have developed several DNA-repair pathways as well as DNA-damage checkpoints to cope with the frequent challenge of endogenous and exogenous DNA insults. In the absence or impairment of such repair or checkpoint mechanisms, the genomic integrity of the organism is often compromised. This review will focus on the functional consequences of impaired DNA-repair pathways. Although each pathway is addressed individually, it is essential to note that cross talk exists between repair pathways, and that there are instances in which a DNA repair protein is involved in more than one pathway. It is also important to integrate DNA-repair process with DNA-damage checkpoints and cell survival, to gain a better understanding of the consequences of compromised DNA repair at both cellular and organismic levels. Functional consequences associated with impaired DNA repair include embryonic lethality, shortened life span, rapid ageing, impaired growth, and a variety of syndromes, including a pronounced manifestation of cancer. PMID- 18285826 TI - ABCA4 gene analysis in patients with autosomal recessive cone and cone rod dystrophies. AB - The ATP-binding cassette (ABC) transporters constitute a family of large membrane proteins, which transport a variety of substrates across membranes. The ABCA4 protein is expressed in photoreceptors and possibly functions as a transporter for N-retinylidene-phosphatidylethanolamine (N-retinylidene-PE), the Schiff base adduct of all-trans-retinal with PE. Mutations in the ABCA4 gene have been initially associated with autosomal recessive Stargardt disease. Subsequent studies have shown that mutations in ABCA4 can also cause a variety of other retinal dystrophies including cone rod dystrophy and retinitis pigmentosa. To determine the prevalence and mutation spectrum of ABCA4 gene mutations in non Stargardt phenotypes, we have screened 64 unrelated patients with autosomal recessive cone (arCD) and cone rod dystrophy (arCRD) applying the Asper Ophthalmics ABCR400 microarray followed by DNA sequencing of all coding exons of the ABCA4 gene in subjects with single heterozygous mutations. Disease-associated ABCA4 alleles were identified in 20 of 64 patients with arCD or arCRD. In four of 64 patients (6%) only one mutant ABCA4 allele was detected and in 16 patients (25%), mutations on both ABCA4 alleles were identified. Based on these data we estimate a prevalence of 31% for ABCA4 mutations in arCD and arCRD, supporting the concept that the ABCA4 gene is a major locus for various types of degenerative retinal diseases with abnormalities in cone or both cone and rod function. PMID- 18285827 TI - Novel SIL1 mutations and exclusion of functional candidate genes in Marinesco Sjogren syndrome. AB - Marinesco-Sjogren syndrome (MSS) is a rare autosomal recessively inherited neurodegenerative disorder characterized by cerebellar ataxia, cataracts, mental retardation, and progressive myopathy. Recently, mutations in the SIL1 gene, which encodes an endoplasmic reticulum (ER) resident cochaperone, were identified as a major cause of MSS. We here report four novel mutations in SIL1, including the first missense substitution p.Leu457Pro described in MSS. In addition, we excluded three functional candidate genes, HSPA5, HYOU1, and AARS, as causative genes in SIL1 mutation-negative patients. To understand the mechanisms of disturbed SIL1 function, we studied the subcellular localization of the missense mutant Leu457Pro protein in COS-1 cells. Moreover, we studied a mutant protein lacking the putative C-terminal ER retrieval signal. In contrast to the wild-type protein's localization to ER and Golgi apparatus, both mutant proteins formed aggregates within the ER depending on the expression level. These data imply that aggregation of mutant proteins may contribute to MSS pathogenesis. The genetic background of a subgroup of patients with MSS remains uncovered. PMID- 18285828 TI - How much is too much? Phenotypic consequences of Rai1 overexpression in mice. AB - The retinoic acid induced 1 (RAI1) gene when deleted or mutated results in Smith Magenis syndrome (SMS), while duplication of 17p11.2, including RAI1, results in the dup(17)(p11.2) syndrome characterized by mental retardation, growth and developmental delays, and hyperactivity. Mouse models for these human syndromes may help define critical roles for RAI1 in mammalian development and homeostasis that otherwise cannot be deduced from patient studies. A mouse model for duplication, Dp(11)17+, involving Rai1 has been reported. However, this mutant was engineered on a mixed genetic background confounding phenotypic effects due to possible modifier genes. We have therefore created and evaluated mice with a graded series of four (hemizygous) and six (homozygous) copies of Rai1, and overexpressing Rai1>1.5-fold and >2-fold, respectively. Data show that Rai1 transgenic mice have growth retardation, increased locomotor activity, and abnormal anxiety-related behavior compared to wild-type littermates. Rai1 transgenic mice also have an altered gait with short strides and long sways, impaired ability on a cage-top hang test, decreased forelimb grip strength, and a dominant social behavior. Further, analyses of homozygous transgenic mice revealed a dosage-dependent exacerbation of the phenotype, including extreme growth retardation, severe neurological deficits, and increased hyperactivity. Our results show that Rai1 dosage has major consequences on molecular processes involved in growth, development, and neurological and behavioral functions, thus providing evidence for several dosage-thresholds for phenotypic manifestations causing dup(17)(p11.2) syndrome or SMS in humans. PMID- 18285829 TI - Pathogenic expansions of the SCA6 locus are associated with a common CACNA1A haplotype across the globe: founder effect or predisposing chromosome? AB - Spinocerebellar ataxia type 6 (SCA6) is a common cause of dominantly inherited ataxia due to an expansion of the CAG repeat in the CACNA1A gene. Affected individuals from the same population share a common haplotype, raising the possibility that most SCA6 cases have descended from a small number of common founders across the globe. To test this hypothesis, we carried out haplotype analysis on SCA6 families from Europe, South America and the Far East, including an established de novo SCA6 expansion. A core CACNA1A disease haplotype was found in affected individuals across the globe. This was also present in the unaffected father of the de novo case, suggesting that the shared chromosome predisposes to the CAG repeat expansion at the SCA6 locus. The SCA6 expansion lies within a CpG island, which could act as a cis-acting element predisposing to repeat expansion as for other CAG/CTG repeat diseases. Polymorphic variation in this region may explain the high-risk haplotype found in SCA6 families. PMID- 18285831 TI - Mapping of a Hirschsprung's disease locus in 3p21. AB - Hirschsprung's disease (HSCR) is a congenital disorder in which ganglion cells are absent in variable portions of the lower digestive tract according to which patients are classified. The RET gene is the major HSCR gene, although reduced penetrance of RET mutations and variable expression of HSCR phenotype indicates that more than one gene is required. An unidentified RET-dependent modifier on 3p21 appears to be necessary for transmission of the short HSCR (S-HSCR) phenotype. We investigated 6 Mb of the 3p21 region on a quest for the HSCR susceptibility locus. Fifty-eight S-HSCR case-parent trios were genotyped using Sequenom technology for 214 tag single nucleotide polymorphisms (SNPs) distributed along 6 Mb of the 3p21 region. A five-marker haplotype, spanning a 118 kb gene-rich region, was found to be overtransmitted to affected offspring. The associated haplotype encompasses three genes involved in neurological phenotypes. Importantly, this association was replicated in an independent sample of 172 S-HSCR cases and 153 unrelated controls. Ranking markers by proximity to candidate genes or by expected functional consequences could be used in follow-up studies to finally pinpoint this HSCR locus. PMID- 18285832 TI - Uptake of testing for BRCA1/2 mutations in South East Scotland. AB - We investigated the uptake of genetic testing by 54 families in South East Scotland with a BRCA1/2 mutation. At a median of 37 months since identification of the mutation, the overall rate of uptake of testing in 269 eligible family members was 32%. First-degree relatives were significantly (P<0.05) less likely to be referred for genetic counselling in more, compared to less, socioeconomically deprived families (46 versus 68%). Among relatives who attended for genetic counselling, females were more likely to be tested than males (76 versus 53%; P<0.05) and relatives with children more than those without children (82 versus 53%; P<0.001). Tested relatives were older than relatives who did not undergo testing (mean 41.9 versus 36.8 years, P<0.05) but did not differ in degree of relationship to the index case or in socioeconomic deprivation. Our results confirm the findings from other studies of substantially lower rates of uptake of genetic testing for BRCA1/2 mutations than anticipated in earlier predictions. Relatives in more socioeconomically deprived families were less likely to be referred for genetic counselling, which is a matter of concern. This may be partly the result of a lack of understanding of the testing process. Cascading currently does not work in breast cancer families and further work is required to investigate intrafamilial communication patterns, testing behaviour and counselling strategies. PMID- 18285833 TI - IFIH1-GCA-KCNH7 locus: influence on multiple sclerosis risk. AB - A recent genome-wide scan of nonsynonymous SNPs and ulterior validation in case control and family analyses evidenced a susceptibility locus for type 1 diabetes (T1D) on chromosome 2q24.3. We aimed at testing the effect of this locus in other autoimmune diseases with complex genetic background, such as multiple sclerosis (MS). Four SNPs along the locus, rs13422767, rs2111485, rs1990760 and rs2068330, were genotyped using TaqMan MGB chemistry in 311 T1D and 412 MS patients and 535 ethnically matched healthy controls. The previously reported association of this locus was found for the first time in MS (rs2068330, G vs C: P=0.001; OR (95% CI)=0.73 (0.6-0.88)) and a trend for replication was observed in our Spanish diabetic cohort. Therefore, genes included in this locus - IFIH1 interferon induced helicase, GCA grancalcin or the potassium channel KCNH7 - are potential candidates implicated in the pathogenesis of these autoimmune diseases, although strong linkage disequilibrium in the region hampered further localization of the etiologic gene. PMID- 18285834 TI - Biallelic mutations in the prokineticin-2 gene in two sporadic cases of Kallmann syndrome. AB - Kallmann syndrome is a developmental disease that combines hypogonadotropic hypogonadism and anosmia. Putative loss-of-function mutations in PROKR2 or PROK2, encoding prokineticin receptor-2 (a G protein-coupled receptor), and one of its ligands, prokineticin-2, respectively, have recently been reported in approximately 10% of Kallmann syndrome affected individuals. Notably, given PROKR2 mutations were found in the heterozygous, homozygous, or compound heterozygous state in patients, thus raising the question of a possible digenic inheritance of the disease in heterozygous patients. Indeed, one of these patients was also carrying a missense mutation in KAL1, the gene responsible for the X chromosome-linked form of Kallmann syndrome. Mutations in PROK2, however, have so far been found only in the heterozygous state. Here, we report on the identification of PROK2 biallelic mutations, that is, a missense mutation, p.R73C, and a frameshift mutation, c.163delA, in two out of 273 patients presenting as sporadic cases. We conclude that PROK2 mutations in the homozygous state account for a few cases of Kallmann syndrome. Moreover, since the same R73C mutation was previously reported in the heterozygous state, and because Prok2 knockout mice exhibit an abnormal phenotype only in the homozygous condition, we predict that patients carrying monoallelic mutations in PROK2 have another disease-causing mutation, presumably in still undiscovered Kallmann syndrome genes. PMID- 18285835 TI - Detection of submicroscopic constitutional chromosome aberrations in clinical diagnostics: a validation of the practical performance of different array platforms. AB - For several decades etiological diagnosis of patients with idiopathic mental retardation (MR) and multiple congenital anomalies (MCA) has relied on chromosome analysis by karyotyping. Conventional karyotyping allows a genome-wide detection of chromosomal abnormalities but has a limited resolution. Recently, array-based comparative genomic hybridization (array CGH) technologies have been developed to evaluate DNA copy-number alterations across the whole-genome at a much higher resolution. It has proven to be an effective tool for detection of submicroscopic chromosome abnormalities causing congenital disorders and has recently been adopted for clinical applications. Here, we investigated four high-density array platforms with a theoretical resolution < or =100 kb: 33K tiling path BAC array, 500K Affymetrix SNP array, 385K NimbleGen oligonucleotide array and 244K Agilent oligonucleotide array for their robustness and implementation in our diagnostic setting. We evaluated the practical performance based on the detection of 10 previously characterized abnormalities whose size ranged from 100 kb to 3 Mb. Furthermore, array data analysis was performed using four computer programs developed for each corresponding platform to test their effective ability of reliable copy-number detection and their user-friendliness. All tested platforms provided sensitive performances, but our experience showed that accurate and user friendly computer programs are of crucial importance for reliable copy-number detection. PMID- 18285837 TI - The success of the genome-wide association approach: a brief story of a long struggle. AB - The genome-wide association approach has been the most powerful and efficient study design thus far in identifying genetic variants that are associated with complex human diseases. This approach became feasible as the result of several key advancements in genetic knowledge, genotyping technologies, statistical analysis algorithms and the availability of large collections of cases and controls. With all these necessary tools in hand, many genome-wide association studies were recently completed, and many more studies which will explore the genetic basis of various complex diseases and quantitative traits are soon to come. This approach has started to reap the fruits of its labor over the past several months. Publications of genome-wide association studies in several complex diseases such as inflammatory bowel disease, type-2 diabetes, breast cancer and prostate cancer have been abundant in the first half of this year. The aims of this review are firstly, to provide a timely summary for most of the genome-wide association studies that have been published until June/July 2007 and secondly, to evaluate to what extent these results have been validated in subsequent replication studies. PMID- 18285836 TI - Pathogenicity of the BRCA1 missense variant M1775K is determined by the disruption of the BRCT phosphopeptide-binding pocket: a multi-modal approach. AB - A number of germ-line mutations in the BRCA1 gene confer susceptibility to breast and ovarian cancer. However, it remains difficult to determine whether many single amino-acid (missense) changes in the BRCA1 protein that are frequently detected in the clinical setting are pathologic or not. Here, we used a combination of functional, crystallographic, biophysical, molecular and evolutionary techniques, and classical genetic segregation analysis to demonstrate that the BRCA1 missense variant M1775K is pathogenic. Functional assays in yeast and mammalian cells showed that the BRCA1 BRCT domains carrying the amino-acid change M1775K displayed markedly reduced transcriptional activity, indicating that this variant represents a deleterious mutation. Importantly, the M1775K mutation disrupted the phosphopeptide-binding pocket of the BRCA1 BRCT domains, thereby inhibiting the BRCA1 interaction with the proteins BRIP1 and CtIP, which are involved in DNA damage-induced checkpoint control. These results indicate that the integrity of the BRCT phosphopeptide-binding pocket is critical for the tumor suppression function of BRCA1. Moreover, this study demonstrates that multiple lines of evidence obtained from a combination of functional, structural, molecular and evolutionary techniques, and classical genetic segregation analysis are required to confirm the pathogenicity of rare variants of disease-susceptibility genes and obtain important insights into the underlying pathogenetic mechanisms. PMID- 18285838 TI - Smith-Lemli-Opitz syndrome: pathogenesis, diagnosis and management. AB - Smith-Lemli-Opitz syndrome (SLOS) is a malformation syndrome due to a deficiency of 7-dehydrocholesterol reductase (DHCR7). DHCR7 primarily catalyzes the reduction of 7-dehydrocholesterol (7DHC) to cholesterol. In SLOS, this results in decreased cholesterol and increased 7DHC levels, both during embryonic development and after birth. The malformations found in SLOS may result from decreased cholesterol, increased 7DHC or a combination of these two factors. This review discusses the clinical aspects and diagnosis of SLOS, therapeutic interventions and the current understanding of pathophysiological processes involved in SLOS. PMID- 18285839 TI - Ceruloplasmin as a marker of occupational copper exposure. AB - Estimation of serum copper to indicate copper status in the human system in the context of moderate chronic occupational copper exposure requires a sophisticated and expensive method. Hence, a search for a suitable marker has been made and few studies have found potential in serum ceruloplasmin. In this context, the present study was initiated to explore whether ceruloplasmin could serve as a predictor of occupational copper exposure. An interviewer-administered questionnaire survey (personal, occupational and health-related information) was undertaken involving 185 employees of a copper handling industry. Serum alkaline phosphatase, serum glutamic pyruvic transaminase (SGPT), serum ceruloplasmin and serum copper were estimated in all the subjects. Multivariate analysis was undertaken using a linear regression model to understand the contribution of serum copper on serum ceruloplasmin values adjusting for the role of other confounders. Serum copper and serum ceruloplasmin values were found to have a statistically significant positive correlation (R=0.169, adjusted R(2)=0.024) after adjustment for other predictors like age, nature of job (department), job duration, smoking, serum alkaline phosphatase and SGPT. This study concludes that the serum ceruloplasmin level can act as a reliable indicator of copper status in the human body following copper exposure in cases of chronic moderate occupational exposure to copper. PMID- 18285841 TI - Patents. PMID- 18285840 TI - Serum PCB concentrations in relation to locally produced food items in eastern Slovakia. AB - Ingestion has been a primary route of PCB exposure for people, especially those not working directly in industrial settings. During 2002-2004, women were recruited at delivery from two districts in eastern Slovakia: Michalovce with high PCB contamination from a chemical manufacturing plant, and Svidnik located 70 km to the northwest, having lower environmental levels of PCBs. Concentrations of 15 PCB congeners were measured in maternal serum using high-resolution gas chromatography with electron capture detection. A food frequency questionnaire was developed and validated at the Research Institute of Nutrition and School of Medicine in Bratislava, Slovakia. The questionnaire was comprised of 88 food items representative of the national dietary habits and designed to ascertain (1) the source of food items as reported by the women (locally produced or purchased from a retail outlet) and (2) quantities consumed of high-fat food categories representative of the national dietary habits. Our primary goal was to identify specific food sources, either locally produced or purchased from retail stores, that might predict serum concentrations of PCBs. We used multiple linear regression to examine the relationship of dietary fats to lipid-adjusted serum PCB levels in 948 adult women (Michalovce N=662, Svidnik N=284) who had recently given birth. We adjusted for residential district, age, body mass index, education and duration of previous lactation. Consumption of fat from locally produced foods was significantly associated with higher levels of lipid-adjusted serum PCB (beta=0.06, P=0.007). Fat from foods purchased in retail outlets showed no significant association (beta=-0.02, P=0.36). There was no interaction between district and diet in predicting serum PCB levels. Comparing women in Michalovce consuming 20 g of fat per day from local sources with those consuming 1 g of fat per day from local sources, lipid-adjusted serum concentrations were predicted to be higher by 81 ng/g lipid or 14.8% 630 ng/g lipid versus 549 ng/g lipid. PMID- 18285842 TI - Analysis and optimization of fabrication of continuous-relief diffractive optical elements. AB - The fabrication of continuous-relief diffractive optical elements by direct laser beam writing in photoresist is analyzed. The main limitation and tolerances are identified, and their influence on optical performance is quantified. Fabricated structures show rounded profile steps resulting from the convolution of the desired profile with the writing beam. This leads to a reduction in diffraction efficiency. Optimization techniques are presented to minimize this effect. Scaling the profile depth by a factor of mu > 1 increases the first-order diffraction efficiency for blazed elements. This method is also applied to suppress the zeroth diffraction order in computer-generated holograms. A nonlinear compensation of the exposure data for the Gaussian beam convolution results in an 18% increase of the diffraction efficiency for a blazed grating with a 10-mum period to a value of 79%. PMID- 18285843 TI - Low-coherence fiber-optic sensor with a large numerical aperture for topographic measurements. AB - We describe a compact fiber-optic setup for three-dimensional topometry. The measuring principle is based on low-coherence interferometry in combination with a confocal microscope. Integrating an optical arrangement based on compact disk optics together with fiber optics into one sensor yields a compact unit. The sensor has a numerical aperture of 0.45 and is suitable for measuring large surface slopes with high spatial resolution. Further, a depth measuring range of more than 1 mm can be achieved. The capabilities of our setup for three dimensional measurements are demonstrated. PMID- 18285844 TI - Reduction of Coupling Loss in a One-to-many Collimating System for a Wavelength Division (de)Multiplexer. AB - In a 1 x N wavelength division (de)multiplexer, N receiving (Rx-) gradient-index rod lenses (GRIN's) are connected to a common transmitting (Tx-) GRIN. All GRIN's are a little longer (DZ(0) for the Tx- and DZ(i) with i = 1, 2, ..., N for the Rx GRIN's) than the quarter-pitch. To reduce the average coupling loss and the deviations, DZ(0) and DZ(i) are optimized independently (unequally) or equally by computer programming for small N, such as N = 4 and 8. For a larger N (e.g., 16), a relay GRIN is required, which is a little (DZ(r)) longer than the half-pitch. The best position of the relay GRIN is located between the seventh and the eighth Rx-GRIN's. Other parameters including DZ(0), DZ(i), and DZ(r) are all optimized. As a result the (de)multiplexer has lower losses. PMID- 18285845 TI - Holographically corrected microscope with a large working distance: errata. PMID- 18285846 TI - Single-shot spatially resolved characterization of laser-induced shock waves in water. AB - We have developed an optical method for single-shot spatially resolved shock-wave peak-pressure measurements. A schlieren technique and streak photography were used to follow the propagation of the shock wave. The shock position r as a function of time was extracted from the streak images by digital image-processing techniques. The resulting r(t) curves were differentiated with respect to time to yield shock-wave velocities that were converted to shock pressures with the aid of the equation of the state of the medium. Features and limitations of the technique are demonstrated and discussed on the basis of measurements of shock wave amplitudes generated by laser-induced breakdown in water. For this purpose, laser pulses of 6-ns duration and pulse energies of 1 and 10 mJ were focused into a cuvette containing water. Complete p(t) curves were obtained with a temporal resolution in the subnanosecond range. The total acquisition and processing time for a single event is ~2 min. The shock-peak pressures at the source were found to be 8.4 ? 1.5 and 11.8 ? 1.6 GPa for pulse energies of 1 and 10 mJ, respectively. Within the first two source radii, the shock-wave pressure p(r) was found to decay on average in proportion to r(-1.3?0.2) for both pulse energies. Thereafter the pressure dropped in proportion to r(-2.2?0.1). In water the method can be used to measure shock-wave amplitudes exceeding 0.1 GPa. Because it is a single-shot technique, the method is especially suited for investigating events with large statistical variations. PMID- 18285847 TI - Optical constants of in situ-deposited films of important extreme-ultraviolet multilayer mirror materials. AB - We have performed angle-dependent reflectance measurements of in situ magnetron sputtered films of B(4)C, C, Mo, Si, and W. The Fresnel relations were used to determine the complex index of refraction from the reflectance data in the region of approximately 35-150 eV. In the cases of Si, C, and B(4)C we found excellent agreement with published data. However, for Mo and W we found that the optical properties from 35 to 60 eV differed significantly from those in the literature. PMID- 18285848 TI - Modified white-light Mach-Zehnder interferometer for direct group-delay measurements. AB - A modified white-light Mach-Zehnder interferometer based on a single beam splitter is described for direct group-delay measurements. The arms of the interferometer are folded in such a manner that a single beam splitter can be used to split the incoming beam and combine the outgoing beams. This method offers a twofold advantage: The measuring range of the interferometer is twice as large as that of the Michelson interferometer, and the systematic error that is associated with the beam splitter is minimized because of the configuration. We report the results of measurements on various optical components performed in the 555-630-nm spectral region and propose a scheme for the processing of the experimental data. We present a comparison of the data analyzed by the proposed processing scheme along with the theoretical calculations. PMID- 18285849 TI - Optical Measurement of Large Sphere Diameters by means of Scattering. AB - A simple method is demonstrated to determine the diameters of dielectric spheres from 0.2 to 1.0 mm by observing the scattering of visible light. Theoretical calculations show that there is an approximately linear relationship between the size of the scattering sphere and the number of maxima and minima in the scattered field as a function of angle when the radius in the 200-1000-wavelength region. PMID- 18285850 TI - Transient deformation analysis by a carrier method of pulsed electronic speckle shearing pattern interferometry. AB - The introduction of a pulsed laser into an electronic speckle-shearing pattern interferometer allows high-speed transient deformations to be measured. We report on a computerized system that permits automatic data reduction by introducing carrier fringes through the translation of a diverging lens. The quantitative determination of the phase map that is due to deformation is carried out by the spatial synchronous detection method. Experimental results obtained for a metal plate transiently deformed by an electromagnetic hammer illustrate the advantages of the proposed system. PMID- 18285851 TI - Liquid refractometry by the rainbow method. AB - A new method for measuring the refractive index of liquid, proposed in a previous paper [Appl. Opt. 36, 5552-5556 (1997)], has been developed. The minimum deviation of a laser beam deflected by a liquid-filled cylindrical cell was calculated by use of geometric optics. These theoretical results were compared with experimental results, with excellent agreement. As a result, the unknown refractive index of a liquid could be obtained by use of a computer calculation to give a best fit. The computer calculation showed that the sensitivity of the refractometer increases with the cell-wall thickness until total reflection takes place. A small refractive-index difference can be detected within a precision of 1 x 10(-6) by use of a metal-oxide semiconductor linear image sensor. We show how to calibrate the refractometer with pure water at 3.98 degrees C. PMID- 18285852 TI - Porosity Depth Profiling of Thin Porous Silicon Layers by use of Variable-Angle Spectroscopic Ellipsometry: A Porosity Graded-Layer Model. AB - Variable-angle spectroscopic ellipsometry was used to determine nondestructively the porosity depth profile and thickness of thin porous silicon layers produced by anodization of p(+)-doped silicon wafers. A porosity graded-layer model is presented and used in the analysis of the material. In the porosity graded-layer model an inhomogeneous layer is built up by several thin sublayers with the porosity changing slightly from one sublayer to the next. Results from the ellipsometry analysis and from transmission electron microscopy reveal inhomogeneous layers whose porosity and thereby optical properties change with their depth in the layers. PMID- 18285853 TI - Common-path optical heterodyne profilometer: a configuration. AB - A novel configuration that combines a linearly polarized He-Ne laser and a birefringent lens to produce a common-path polarized optical heterodyne profilometer with respect to the heterodyned P and S waves has been set up. In this profilometer a linear polarized frequency-stabilized He-Ne laser was used with an acousto-optical modulator to replace the Zeeman laser as the light source that had two polarization eigenstates in different temporal frequencies. The proposed interferometer shows a more symmetric and ideal common-path structure than the conventional optical heterodyne profilometers with the Zeeman laser. The phase error aroused by the elliptical polarization and the nonorthogonality of the two eigenpolarization modes of the Zeeman laser can be reduced. The system's resolution in the vertical direction reaches 2 A, and in a 27-mum scanning range the repeatability of the surface profile measurements is shown to be 5 A. PMID- 18285854 TI - Magnification of conic mirror reflectometers. AB - Conic mirror reflectometers are used to measure the diffuse reflectance and total integrated scatter of surfaces. In spite of the long history of using conic mirrors for these purposes, the maximum magnification of the three primary types of conic mirror (hemisphere, hemiellipsoid, and dual paraboloid) had not been compared quantitatively. To our knowledge, an exact magnification formula has not been published for any of the three primary conic mirrors. The maximum magnification is needed for proper sizing of detectors and radiation sources used with reflectometers. Exact analytical expressions for the maximum magnification of a Coblentz hemisphere, a hemiellipsoid, and a dual-paraboloid mirror system are derived and compared. PMID- 18285855 TI - Fringe Formation in an In-Plane Displacement Measurement Configuration with Twofold Increase in Sensitivity: Theory and experiment. AB - A fringe-formation theory for a dual-beam illumination configuration that leads to a twofold increase in sensitivity for the measurement of in-plane displacement is described. Here we have taken into account all four beams simultaneously that are generated at the image plane owing to two-beam illumination and their cross interference terms for fringe formation. We show that the sensitivity obtainable is the usual interferometric sensitivity when we take into account all four beams simultaneously and doubles only when the retroreflected beams are observed. A detailed theory and an experimental demonstration of the method are presented. PMID- 18285856 TI - Nanoradian angle sensor and in situ self-calibration. AB - We describe a high-precision angle sensor and a new calibration system for calibrating the linearity error of an angle sensor under in situ conditions without the need for accurate reference instruments. To achieve accuracy in the nanoradian (nrad) order, we elongated the critical-angle prism to increase the sensitivity, and we added a thermal controller to the angle sensor. This served to suppress thermal drift of the light source to improve stability. The proposed angle sensor demonstrated a stability of 3 nrad over 1 min, and the maximum repeatability error of the calibration curve determined by the in situ self calibration method was approximately 35 nrad with a measurement range of 500 murad. PMID- 18285857 TI - Simple Method for Determining Slowly Varying Refractive-Index Profiles from in situ Spectrophotometric Measurements. AB - Reliable control of the deposition process of optical films and coatings frequently requires monitoring the refractive-index profile throughout the layer. In the present research a simple in situ approach is proposed that uses a WKBJ matrix representation of the optical transfer function of a single thin film on a substrate. Mathematical expressions are developed that represent the minima and the maxima envelopes of the curves transmittance versus time and reflectance versus time. The refractive index and the extinction coefficient depth profiles of different films are calculated from simulated spectra as well as from experimental data obtained during the PECVD (plasma-enhanced chemical vapor deposition) of silicon-compound films. Variation in the deposition rate with time is also evaluated from the position of the spectra extrema as a function of time. The physical and mathematical limitations of the method are discussed. PMID- 18285858 TI - 1 x N star coupler as a distributed fiber-optic strain sensor in a white-light interferometer. AB - A novel technique of using a 1 x N star fiber optic coupler as a distributed strain sensor in a white-light interferometer to measure the distribution of strain is presented. The measuring principle and 1 x 4 star coupler with four fiber optic strain sensors are demonstrated. The experiment is performed with four sensors attached to a combination plastic specimen. PMID- 18285859 TI - Numerical simulations of scattering in a two-photon optical data storage system. AB - Scattering effects in a two-photon optical data storage system are numerically studied. Surface scattering analysis with a scalar, beam propagation model is performed. We analyze the problem by modeling scattering from randomly varying surfaces and also by Fourier surface decomposition. Scattering induced by propagation through multiple pages of randomly recorded data marks is also studied with a hybrid finite-difference-time-domain/angular-spectrum model. Both surface and bulk scattering are shown to influence the spatial properties of the optical beam. Results and some possible implications are presented. PMID- 18285860 TI - High-resolution inset head-mounted display. AB - A novel approach to inset superimposition in a high-resolution head-mounted display (HMD) is presented. The approach is innovative in its use of optoelectronic, nonmechanical devices in place of scanning mechanical devices commonly adopted previously. A paraxial layout of the overall HMD system is presented, and the benefit of employing hybrid refractive-diffractive optics for the optical component that generates the inset is discussed. A potential overall HMD design is finally presented to show the integrated system. The practical limitations of the designed system are discussed and an alternative approach is presented to compare the advantages and the limitations of these systems. PMID- 18285861 TI - Elliptical trough reflector for the collection of light from linear sources. AB - A trough reflector with a reflective, truncated elliptical surface was designed to efficiently collect freely propagating light from a linear source. The source was placed at one focus of the reflector, and light was collected through a rectangular aperture near the second focus. Collection efficiency was much greater than that of a spherical integrator and approximately 6.5x greater than that of an objective lens; as much as approximately 55% of the light could be captured from the full aperture. This reflector could be used to efficiently collect surface fluorescence excited by use of evanescent waves in fluorescence based fiber optic or capillary waveguide sensors. PMID- 18285862 TI - Aberrated lenses for generating flattened laser irradiance. AB - Expressions for describing Gaussian beams focused by a lens with spherical aberration have been derived. Numerical results show that, when the coefficient of the spherical aberration is negative, one can obtain flattened laser irradiance at two positions along the focused field. The larger the coefficient of negative spherical aberration, the larger the flatness is. The effect of the Fresnel number of the focusing lens on the flattened laser irradiance is also investigated. PMID- 18285863 TI - Two-Element Refracting System for Annular Gaussian-to-Bessel Beam Transformation. AB - A refracting system consisting of two lenses is designed to transform an annular Gaussian laser beam into a circular Bessel beam. The slopes of the input and output surfaces fit well with a sixth-order polynomial. A smooth variation of the radii of curvature of the resulting aspheric surfaces is very attractive for easy machining of the surfaces. The diffraction-free length for the designed system is 59.4735 m at 633 nm. PMID- 18285864 TI - Bicell Pyroelectric Optical Detector Made from a Single LiNbO(3) Domain-Reversed Electret. AB - Using electric-field poling at room temperature, we selectively reversed the direction of the spontaneous polarization in a 200-mum-thick, z-cut LiNbO(3) electret to produce a bicell pyroelectric detector. The detector required only a single set of electrodes, one electrode on the front surface and one on the back surface. Microphonic noise that is typical of monocell pyroelectric detectors is reduced in the present device. Our spatial uniformity data indicate that the optical response of one half of the bicell detector area was equal to and opposite the other half within 1.2%. The microphonic suppression of the bicell pyroelectric detector was less than -36 dB from 10 to 50 Hz and less than -118 dB at 35 Hz of that of a reference monocell pyroelectric detector. The substrate thickness is significantly greater than those of other domain-engineered pyroelectric detector designs and allows us to build practical, large-area detectors for radiometric applications. PMID- 18285865 TI - Thin-film biological reflectors: optical characterization of the Chrysiridia croesus moth. AB - The optical properties of colored wing scales of the Chrysiridia croesus moth were investigated experimentally and theoretically by reflection spectroscopy and ellipsometry. Transmission electron microscope micrographs show that the outer surfaces of these scales incorporate a fairly regular layered structure of alternating dense and less-dense material, which reflects light by the well-known thin-film interference process. A Monte Carlo-type simulation of the reflection process is discussed, which permits the determination of the complex index of refraction of the scale material. PMID- 18285866 TI - Integral localized approximation in generalized lorenz-mie theory. AB - The generalized Lorenz-Mie theory deals with the interaction between spheres and arbitrarily shaped illuminating beams. An efficient use of the theory requires efficient evaluation of the so-called beam-shape coefficients involved in the description of the illuminating beam. A less time-consuming method of evaluation relies on the localized approximation. However, it lacks flexibility when the description of the illuminating beam is modified. We present a new version of this method, called the integral localized approximation, that exhibits the desired property of flexibility. PMID- 18285867 TI - Artificial diamond as a broadband infrared beam splitter for fourier transform spectroscopy. AB - We show that an artificially grown diamond plate serves as a very broadband beam splitter from approximately 7 mum, where the intrinsic diamond absorption becomes important, to the submillimeter region (a 250-mum wavelength or longer). PMID- 18285868 TI - Real-time characterization of film growth on transparent substrates by rotating compensator multichannel ellipsometry. AB - A multichannel spectroscopic ellipsometer in the fixed-polarizer-sample-rotating compensator-fixed-analyzer (PSC(R)A) configuration has been developed and applied for real-time characterization of the nucleation and growth of thin films on transparent substrates. This rotating-compensator design overcomes the major disadvantages of the multichannel ellipsometer in the rotating-polarizer-sample fixed-analyzer (P(R)SA) configuration while retaining its high speed and precision for the characterization of thin-film processes in real time. The advantages of the PSC(R)A configuration include (i) its high accuracy and precision for the detection of low-ellipticity polarization states that are generated upon reflection of linearly polarized light from transparent film substrate systems, and (ii) the ability to characterize depolarization of the reflected light, an effect that leads to errors in ellipticity when measured with the P(R)SA configuration. A comparison of the index of refraction spectra for a glass substrate obtained in the real-time PSC(R)A mode in 2.5 s and in the ex situ fixed-polarizer-fixed-compensator-sample-rotating-analyzer (PCSA(R)) mode in ~10 min show excellent agreement, with a standard deviation between the two data sets of 8 x 10(-4), computed over the photon energy range from 1.5 to 3.5 eV. First, we describe the PSC(R)A ellipsometer calibration procedures developed specifically for transparent substrates. In addition, we describe the application of the multichannel PSC(R)A instrument for a study of thin-film diamond nucleation and growth on glass in a low-temperature microwave plasma-enhanced chemical vapor deposition process. PMID- 18285869 TI - Design and Manufacture of Low-Absorption ZnS-YF(3) Antireflection Coatings in the 3.5-16-num Spectral Range. AB - Broadband antireflection coatings on ZnSe substrates are designed and manufactured in the 3.5-16-mum IR range. The thin-film materials are YF(3) and ZnS produced by electron-beam deposition. To reach optimal performances to as great as 16 mum, we performed an accurate determination of complex indices, taking into account dispersion laws and absorption due to water within layers. With this knowledge, designs are optimized and absorption is reduced at 16 mum. PMID- 18285870 TI - Optical waveguiding in epitaxial PbTiO(3) thin films. AB - Epitaxial lead titanate (PbTiO(3)) thin films on SrTiO(3) (100) substrate were grown in situ by radio-frequency sputtering for optical waveguiding applications. The crystalline quality of the PbTiO(3) films deposited at 550 degrees C has been investigated through x-ray diffraction analysis. It indicates that thin films are completely c-axis oriented (rocking curve FWHM of 0.2 degrees for the 001 reflection). The transmission spectrum method has been used to measure the dispersion of the refractive index. At 632.8 nm, the PbTiO(3) film with an (001) orientation exhibits a refractive index of 2.61, which represents 98% of the bulk material. The prism-coupling technique has been also employed to determine the optical attenuation in the planar waveguide. In this study, we report a low propagation loss of 2.2 ? 0.2 dB/cm obtained in a PbTiO(3) optical waveguide. PMID- 18285871 TI - Narrow-band reflector coatings: design procedures. AB - A class of problem dealing with narrow-band reflectors under oblique light incidence has been selected, and a group of design procedures leading to satisfactory results is analyzed. Initially performance criteria and design restrictions are set. Various designs are then created and analyzed. Conclusions can then be drawn on the relative merits of the designs. Two different types of target function have been chosen. A global search for 50 or more layers with varied optical thicknesses between 0 and lambda/2 and a needle design method have been applied to the synthesis. Gradient and variable metrics have been used for further refinement. Results are compared, and it is shown that all the design methods used yield similar results for this problem. The issue of how to determine the best method is addressed. PMID- 18285872 TI - Attenuated phase-shifting masks of chromium aluminum oxide. AB - Chromium aluminum oxide was chosen as a new candidate for use as an attenuated phase-shifting mask (Att-PSM) material. The compositions of films were correlated with optical properties. With the measured and the fitted data, we simulated the transmittance and the phase shift using the matrix method. Consequently, we acquired optimum parameters for Att-PSM's, such as Al/Cr = 1.9-2.5 and d = 120 nm at a 193-nm wavelength, Al/Cr = 1.0-1.7 and d = 128 nm at a 248-nm wavelength, and Al/Cr = 0-0.1 and d = 170 nm at a 365-nm wavelength. This simulation was verified by transmittance measurement. PMID- 18285873 TI - Analysis of errors in thin-film optical parameters derived from spectrophotometric measurements at normal light incidence. AB - A comparative analysis is made of the errors in deriving the optical parameters (n, refractive index; k, absorption coefficient; d, film thickness) of thin films from spectrophotometric measurements at normal light incidence. The errors in determining n, k, and d by the (TR(f)R(b)), (TR(f)R(m)), (TR(b)R(m)), (TR(f)), (TR(m)), and T(k = 0) methods are compared. It is shown that they are applicable to optical constants of thin films in the n > 1.5, k < 4.5, and d/lambda = (0.02 0.3) range, and their combinations make possible the determination of n and k to an accuracy of better than ?4%. To derive the optical constants in a wide spectral range with high accuracy and isolate the correct physical solutions reliably, one should apply all methods, using the relevant solutions with the lowest errors, as shown in this research, when determining the optical constants of As(2)S(3) and Sb(2)Se(3) films. PMID- 18285874 TI - Generalized abeles relations for an anisotropic thin film with an arbitrary dielectric tensor: comments. AB - The Cojocaru generalization of the 2 x 2 extended Jones matrix method, placed in a wider context of previous approaches to anisotropic optical thin films, is analyzed from a complementary perspective. This, contrary to initial belief, allows for a simple proof that one may include multiple reflections by taking into account total fields into the anisotropic film, and this therefore provides support for a more widespread use of the method. PMID- 18285875 TI - Chromospheric Helium Imaging Photometer (an Instrument for High Time Cadence 1083 nm Wavelength Solar Observations). AB - The scientific motivation, design criteria, and specifications for a new ground based instrument to observe the Sun in the He i 1083-nm spectral line is described. The instrument employs a liquid-crystal tunable Lyot-type spectral filter and an array detector that allows the full solar disk to be observed with a time cadence of minutes. We describe the telescope's optical and mechanical features and discuss computer interface and data-reduction procedures employed. Instrument performance during the initial year of operation of the telescope at its high-altitude site is summarized. PMID- 18285876 TI - Baffling and Shielding System for a Millimeter-Wave balloonborne Telescope. AB - We describe a balloonborne telescope devoted to millimeter observations of diffuse sky radiation and optimized with a custom set of baffles and shields. The basic idea of the shield design is to shape the surfaces as roof mirrors to redirect the stray radiation in the opposite direction of incidence. The baffles are used to stop radiation close to the field of view. In this way we are able to avoid most of the unwanted radiation. We successfully operated this telescope (ARGO) during two flights, in 1989 and 1993, while detecting the faint structures of the cosmic microwave background at degree scale. PMID- 18285877 TI - Very-wide-angle optical systems suitable for spaceborne photometric measurements. AB - Spaceborne visible-light images for observing the large angular extent of the solar corona require 0.1% differential broadband photometry over ~1 degrees sky bins. When we are using a CCD camera, this specification requires spreading unresolved images over many pixels. Large images ease correction for aberration or field curvature. Permitting large images allows simple and lightweight very wide-angle designs employing spherical and toroidal mirrors and thick lenses that can view almost the entire sky. We present formulas and graphic results relating sky angle to focal-plane position and determining the tangential and sagittal focal surfaces governing image size at the CCD. Laboratory measurements with two prototype configurations confirm the calculations. PMID- 18285878 TI - Two-dimensional inverse problem of diffusion tomography: the approach applicable for small inclusions. AB - A method for solving the two-dimensional inverse problems of optical diffusion tomography is proposed. The method is especially designed for the imaging of small inclusions embedded in the backgrounds of strongly scattering media. Numerical simulations show that the results are stable with respect to external noise at the boundary of the sample. The location of an inclusion is obtained with an accuracy of the order of several photon transport mean-free paths in the medium in cases both with and without noise in the scattering data used for the solution of the inverse problem. PMID- 18285879 TI - Resolution improvement in microscopic imaging through turbid media based on differential polarization gating. AB - We report a new method for microscopic imaging of an object embedded in a turbid medium, based on the differential polarization-gating mechanism. It is demonstrated that with this method, image resolution through optically thick milk suspensions can be improved by as much as 30% compared with no-gating methods. An image resolution of tens of micrometers is achieved in an optically thick turbid medium, which is approximately 10 times better than that achieved in transillumination imaging in a similar medium. PMID- 18285880 TI - Simple, low-cost, portable corneal fluorometer for detection of the level of diabetic retinopathy. AB - A simple, low-cost, portable instrument for measurement of the autofluorescence of the human cornea is presented. Corneal autofluorescence has proved to be strongly correlated with the grade of retinopathy in diabetic patients. It is therefore a reliable parameter for detection of different levels of diabetic retinopathy, thus permitting timely intervention by ophthalmologists. The instrument contains custom optics and electronics and exhibits excellent linearity and repeatability both in vitro and in vivo. Preliminary tests on volunteers show promise for its use in clinical practice. PMID- 18285881 TI - Algorithm to increase the largest aberration that can be reconstructed from Hartmann sensor measurements. AB - Conventional Hartmann sensor processing relies on locating the centroid of the image that is formed behind each element of a lenslet array. These centroid locations are used for computing the local gradient of the incident aberration, from which the phase of the incident wave front is calculated. The largest aberration that can reliably be sensed in a conventional Hartmann sensor must have a local gradient small enough that the spot formed by each lenslet is confined to the area behind the lenslet: If the local gradient is larger, spots form under nearby lenslets, causing a form of cross talk between the wave-front sensor channels. We describe a wave-front reconstruction algorithm that processes the whole image measured by a Hartmann sensor and a conventional image that is formed by use of the incident aberration. We show that this algorithm can accurately estimate aberrations for cases in which the aberration is strong enough to cause many of the images formed by individual lenslets to fall outside the local region of the Hartmann sensor detector plane defined by the edges of a lenslet. PMID- 18285882 TI - Photoetching cellulose-film recording material and fabrication of double-faced micro-optical elements. AB - A novel, to our knowledge, polymer-type holographic and photoetching recording material-photoetching cellulose film-is reported. Its basic photochemical reaction scheme and the mechanism of image formation are discussed with respect to electron-spin resonance spectra and infrared spectra. Meanwhile, its characteristics, such as a strong real-time effect, high spatial resolution, linear surface-relief modulation capacity, and delayed development effect, are investigated experimentally. Finally, taking advantage of its double-faced photosensitivity, we fabricate integrated micro-optical elements that have both the functions of beam splitting and focusing. PMID- 18285883 TI - Reconfigurable two-dimensional diffractive phase element with the fractional talbot effect. AB - We investigate the technical feasibility of the optical implementation of a four phase-level diffractive element with two {0, pi}-phase spatial light modulators in a fractional Talbot configuration. The space-bandwidth product of the spatial light modulators is seen as the main theoretical limitation of the proposed approach. We investigate the robustness of technological and geometrical parameters on the diffraction efficiency of the whole system. Ferroelectric liquid-crystal silicon backplane spatial light modulators are chosen because of their high reconfiguration rates and good electro-optics interface. Similarly, we assess the influence of liquid-crystal technical parameters on system performance. PMID- 18285884 TI - High-Contrast Spatial Light Modulator by use of the Electroabsorption and the Electro-Optic Effects in a GaAs Single Crystal. AB - A new spatial light modulator that uses the electroabsorption and the electro optic effects in a GaAs single crystal is proposed. The device has the same structure as a Pockels readout optical modulator and can be operated at a frame rate higher than 500 Hz. When the electroabsorption and the electro-optic effects are combined, the dynamic range (contrast ratio) becomes larger than that which results when either effect is used singly. It was experimentally confirmed that the modulator has a high contrast ratio (greater than 2000:1), high sensitivity, and consequently large gamma characteristics. PMID- 18285885 TI - High-performance talbot array illuminators. AB - The performance of two-dimensional Talbot array illuminators is discussed in terms of compression ratio, fabrication cost, and illumination efficiency. By comparing two array-illuminator families, we try to answer the question, "Which Talbot array illuminator provides the best illumination performance and requires the least expenditure in fabrication for a given compression ratio?" We further present experimental results obtained with a quartz-glass four-level surface relief array illuminator designed for a two-dimensional compression ratio of 16 and fabricated with only two lithographic masks. PMID- 18285886 TI - White-light-modified Talbot array illuminator with a variable density of light spots. AB - A flexible array illuminator, comprising only two conventional optical elements, with a variable density of bright white-light spots is presented. The key to our method is to obtain with a single diffractive lens an achromatic version of different fractional Talbot images, produced by free-space propagation, of the amplitude distribution at the back focal plane of a periodic refractive microlens array under a broadband point-source illumination. Some experimental results of our optical procedure are also shown. PMID- 18285887 TI - Generalized Wigner function for the analysis of superresolution systems. AB - The generalized Wigner function is able to represent light distributions that contain spatial and temporal information. The use of such a generalized Wigner distribution function for analysis and understanding of temporally restricted superresolving systems is demonstrated. These systems gain spatial resolution by conversion of the temporal degrees of freedom to spatial degrees of freedom. PMID- 18285888 TI - Compact phase-conjugating correlator: simulation and experimental analysis. AB - A simulation and experimental investigation of a recently proposed, compact, phase-conjugating correlator is undertaken. The effects of noise and other distortions in the input image and in the correlator filter plane are considered. As with other phase-only designs, the phase-conjugating correlator is sensitive to distortion of the input image while being robust in the presence of filter plane distortions; this robustness is enhanced by the phase-conjugating property of the design. PMID- 18285889 TI - Four-Channel, 8 x 8 Bit, Two-Dimensional Parallel Transmission by use of Space Code-Division Multiple-Access Encoder and Decoder Modules. AB - We experimentally demonstrate four-channel multiplexing of 64-bit (8 x 8) two dimensional (2-D) parallel data links on the basis of optical space-code-division multiple access (CDMA) by using new modules of optical spatial encoders and a decoder with a new high-contrast 9-m-long image fiber with 3 x 10(4) cores. Each 8 x 8 bit plane (64-bit parallel data) is optically encoded with an 8 x 8, 2-D optical orthogonal signature pattern. The encoded bit planes are spatially multiplexed and transmitted through an image fiber. A receiver can recover the intended input bit plane by means of an optical decoding process. This result should encourage the application of optical space-CDMA to future high-throughput 2-D parallel data links connecting massively parallel processors. PMID- 18285890 TI - Scalar diffraction theory approach to estimating multimode-waveguide field amplitude mode distributions. AB - We introduce a method to estimate the coupling coefficients of the guided field amplitude and the corresponding angular bandwidth in a multimode slab waveguide. This scalar diffraction theory approach is simpler than the more rigorous electromagnetic treatment and is directly applicable to communications systems that use large (dimensions or numerical aperture) waveguides, as in substrate mode interconnects. Moreover, this method provides conceptual insight as to a parameter's effect on the field-amplitude mode distribution and angular bandwidth. PMID- 18285891 TI - Recoded and nonrecoded trinary signed-digit adders and multipliers with redundant bit representations. AB - Highly-efficient two-step recoded and one-step nonrecoded trinary signed-digit (TSD) carry-free adders-subtracters are presented on the basis of redundant-bit representation for the operands' digits. It has been shown that only 24 (30) minterms are needed to implement the two-step recoded (the one-step nonrecoded) TSD addition for any operand length. Optical implementation of the proposed arithmetic can be carried out by use of correlation- or matrix-multiplication based schemes, saving 50% of the system memory. Furthermore, we present four different multiplication designs based on our proposed recoded and nonrecoded TSD adders. Our multiplication designs require a small number of reduced minterms to generate the multiplication partial products. Finally, a recently proposed pipelined iterative-tree algorithm can be used in the TSD adders-multipliers; consequently, efficient use of all available adders can be made. PMID- 18285892 TI - Fluorescence readout of near-field photochromic memory. AB - The fluorescence- and transmittance-detection readout methods of a near-field photochromic optical memory by a scanning-probe technique were studied theoretically. Shot noise, as the principal noise, was taken into account in an analysis of the signal-to-noise ratio (SNR). Under most conditions, fluorescence detection readout yielded a better SNR than did transmittance-detection readout. For a transmittance change of 0.9-1.0 as a result of recording, a readout light power of approximately 100 nW, and a system bandwidth of 1 MHz, only the fluorescence-detection readout method, under the condition that the fluorescent layer of the medium have a fluorescence quantum yield greater than 0.4, can produce a sufficiently large SNR. PMID- 18285893 TI - Measurements and simulations of differential phase-tracking signals in optical disk data storage. AB - The tracking-error signal generated in differential phase detection (DPD) is theoretically analyzed and numerically simulated. Experimental measurements of the DPD signal versus the tracking offset obtained on compact read-only and phase change disks are also reported. The signal is sensitive to the geometry of the marks, intersymbol interference along the track, and cross-track cross talk. A characteristic parameter is introduced to relate the DPD signal to the reflectivities of the mark and the spacer. For read-only disks such as CD-ROM and DVD-ROM, the magnitude of the DPD signal does not seem to depend on the reflectivity of the disks, nor does it depend on the pit depth. As for the influence of the various aberrations on the DPD signal, coma in the cross-track direction is shown to give rise to significant tracking offset, whereas defocus and spherical aberrations reduce the magnitude of the DPD signal appreciably. PMID- 18285894 TI - Power-law correlations in natural infrared imagery. AB - We provide experimental data confirming the power-law spatial-correlation model of radiance statistics for a wide range of natural terrains and in two IR wave bands. We analyze imagery of four terrains that was acquired from an airborne sensor in both midwave and long-wave bands. Data are presented on the region of the correlation-model parameter space occupied by natural terrains. PMID- 18285895 TI - Wavelet-based image enhancement in x-ray imaging and tomography. AB - We consider an application of the wavelet transform to image processing in x-ray imaging and three-dimensional (3-D) tomography aimed at industrial inspection. Our experimental setup works in two operational modes-digital radiography and 3-D cone-beam tomographic data acquisition. Although the x-ray images measured have a large dynamic range and good spatial resolution, their noise properties and contrast are often not optimal. To enhance the images, we suggest applying digital image processing by using wavelet-based algorithms and consider the wavelet-based multiscale edge representation in the framework of the Mallat and Zhong approach [IEEE Trans. Pattern Anal. Mach. Intell. 14, 710 (1992)]. A contrast-enhancement method by use of equalization of the multiscale edges is suggested. Several denoising algorithms based on modifying the modulus and the phase of the multiscale gradients and several contrast-enhancement techniques applying linear and nonlinear multiscale edge stretching are described and compared by use of experimental data. We propose the use of a filter bank of wavelet-based reconstruction filters for the filtered-backprojection reconstruction algorithm. Experimental results show a considerable increase in the performance of the whole x-ray imaging system for both radiographic and tomographic modes in the case of the application of the wavelet-based image processing algorithms. PMID- 18285896 TI - Edge enhancement by use of moving gratings in a bismuth silicon oxide crystal and its application to optical correlation. AB - The technique of moving gratings in a photorefractive crystal is applied to the edge enhancement of objects and edge-enhanced optical correlation. The nonlinear dependence of the optimum fringe velocity on the fringe modulation and the variation of the enhancement of the diffraction efficiency with fringe modulation at a fixed fringe velocity appropriate to high fringe modulations are experimentally investigated. It is shown that the diffraction at high fringe modulations, which corresponds to the high-spatial-frequency components of the Fourier spectrum, is enhanced by a factor of approximately 3.7, whereas the diffraction at low fringe modulations is suppressed by a factor of 0.6. The proposed technique has the advantages of real-time enhancement, arbitrary selection of the spatial frequency to be enhanced, and improved stability of the output. Experimental results of the edge enhancement of objects and edge-enhanced correlation are presented. PMID- 18285897 TI - Optimal Wave-front Reconstruction for a Coherent Diffracted Field. AB - Phase differences in the far field of a coherently illuminated object are used to estimate the two-dimensional phase in the measurement plane of an imaging system. A previously derived phase-correlation function is used in a minimum-variance phase-estimation algorithm to map phase-difference measurements optimally to estimates of the phase on a grid of points in the measurement plane. Theoretical and computer-simulation comparisons between the minimum-variance phase estimator and conventional least-squares estimators are made. The minimum-variance phase estimator produces a lower aperture-averaged mean-square phase error for all values of a sampling parameter beta. PMID- 18285898 TI - Robust phase-unwrapping algorithm with a spatial binary-tree image decomposition. AB - The search for fast and robust phase-unwrapping algorithms remains an important problem in the development of real-time interferometric systems. Our phase unwrapping approach uses a spatial binary-tree image decomposition to permit maximum parallelism in implementation. At each node in the tree structure, a single unwrapping decision is made between two image blocks. The unwrapping rule is derived from a statistical-estimation framework. Specifically, a maximum likelihood estimate of the demodulation term is used. This term can be viewed as that which minimizes a discontinuity-penalizing cost function. We show that the algorithm exhibits a high level of robustness. Quantitative measures of performance are provided, and many phase maps are shown for subjective evaluation. PMID- 18285899 TI - Adaptive-neighborhood filtering of images corrupted by signal-dependent noise. AB - In many image-processing applications the noise that corrupts the images is signal dependent, the most widely encountered types being multiplicative, Poisson, film-grain, and speckle noise. Their common feature is that the power of the noise is related to the brightness of the corrupted pixel. This results in brighter areas appearing to be noisier than darker areas. We propose a new adaptive-neighborhood approach to filtering images corrupted by signal-dependent noise. Instead of using fixed-size, fixed-shape neighborhoods, statistics of the noise and the signal are computed within variable-size, variable-shape neighborhoods that are grown for every pixel to contain only pixels that belong to the same object. Results of adaptive-neighborhood filtering are compared with those given by two local-statistics-based filters (the refined Lee filter and the noise-updating repeated Wiener filter), both in terms of subjective and objective measures. The adaptive-neighborhood approach provides better noise suppression as indicated by lower mean-squared errors as well as better retention of edge sharpness than the other approaches considered. PMID- 18285900 TI - Method of error analysis for phase-measuring algorithms applied to photoelasticity. AB - We present a method of error analysis that can be applied for phase-measuring algorithms applied to photoelasticity. We calculate the contributions to the measurement error of the different elements of a circular polariscope as perturbations of the Jones matrices associated with each element. The Jones matrix of the real polariscope can then be calculated as a sum of the nominal matrix and a series of contributions that depend on the errors associated with each element separately. We apply this method to the analysis of phase-measuring algorithms for the determination of isoclinics and isochromatics, including comparisons with real measurements. PMID- 18285901 TI - Theoretical and experimental study of the bleaching of a dye in a film polymerization process. AB - The quantum efficiency and the molar-absorption coefficients of different phenothiazine dyes are obtained by means of fitting the experimental data of transmittance as a function of time. An analytical expression for the intensity transmitted in a photopolymerizable holographic material is obtained, and good agreement between theory and experience is also achieved. The analysis of these parameters is of fundamental quantities in the photochemical characterization of holographic recording materials. PMID- 18285902 TI - Beam shaping and its solution with the use of an optimization method. AB - We present an exact mathematical description of beam shaping and indicate that a rigorous solution does not exist: only an optimal solution can be found. An optimization method is proposed to search for the solution. The simulation results for an example are given in detail. PMID- 18285903 TI - Generalized spin filtering and an improved derivative-sign binary image method for the extraction of fringe skeletons. AB - Generalized spin filters, including several directional filters such as the directional median filter and the directional binary filter, are proposed for removal of the noise of fringe patterns and the extraction of fringe skeletons with the help of fringe-orientation maps (FOM's). The generalized spin filters can filter off noise on fringe patterns and binary fringe patterns efficiently, without distortion of fringe features. A quadrantal angle filter is developed to filter off the FOM. With these new filters, the derivative-sign binary image (DSBI) method for extraction of fringe skeletons is improved considerably. The improved DSBI method can extract high-density skeletons as well as common density skeletons. PMID- 18285904 TI - Adaptive optics: introduction to the feature issue. PMID- 18285905 TI - Density of turbulence-induced phase dislocations. AB - Under certain conditions, light-wave propagation through turbulent media causes a specific type of phase distortion: so-called phase dislocations. A salient feature of phase dislocations is an appearance of zones where the phase turns out to be a multivalued function of coordinates. The problem of turbulence-induced phase dislocations is considered. Both a theoretical treatment and simulations based on the numerical solution of a parabolic equation are used for estimation of the dislocation density. Various turbulence conditions, ranging from weak to very strong ones, are considered as well as the dependences on wavelength, and the inner scales of turbulence are presented. An empirical formula for the dislocation density suitable for a wide range of turbulent and propagation conditions is derived. The results obtained can be useful for both atmospheric and adaptive optics. PMID- 18285906 TI - Potential and vortex features of optical speckle fields and visualization of wave front singularities. AB - The feasibility of noninterferometric methods to measure phase distribution in a laser beam cross section for visualization of the vortex dislocations of an optical speckle-field wave front is analyzed. Peculiarities of the phase retrieved from the measured intensity distribution (the phase problem in optics) and from the wave-front slopes measured by a Hartmann sensor are discussed. A concept of the vortex and the potential parts of the phase is introduced. An analytic formula to retrieve the potential phase from the measured intensity has been obtained. We show that the considered means of measurements allow the positions of the dislocation centers to be sensed and the spatial configuration of the intensity zero lines to be reconstructed. PMID- 18285907 TI - Finite temporal measurements of the statistical characteristics of the atmospheric coherence length. AB - Based on the theory of stationary random processes, the probability density function of the atmospheric coherence length measured in a finite time period is derived analytically and verified experimentally. An iteration method is proposed to obtain Fried's coherence length within a finite time measurement. PMID- 18285908 TI - Modal compensation and the atmospheric-turbulence outer scale: computer simulations. AB - The influence of the turbulence outer scale on the Strehl ratio obtained with low order adaptive optics systems is examined by numerical simulation. The Karhunen Loeve approach is used to generate wave-front samples. A method that allows construction of the outer-scale-dependent Karhunen-Loeve functions is described. It is shown that the Strehl ratio produced by a second-order adaptive optics correction (tip-tilt, defocus, and astigmatism) is affected quite strongly by the finite outer scale. For the higher-order correction, the effect under study is weak and appears only when the outer-scale magnitude becomes less than the aperture diameter. It is also shown that the finite outer scale has a positive effect on the Strehl ratio of the uncorrected long-exposure image. PMID- 18285909 TI - Phase-compensation experiment with a 37-element adaptive optics system. AB - An experimental study on phase compensation for turbulent effects with a 37 element adaptive optics system is performed in both a simulated turbulence cell and in a real atmosphere. The experimental results demonstrate that the compensated Strehl ratio S(0), which is influenced mainly by the deformable mirror fitting error, has a functional form S(0) = exp[-kappa(d/r(0))(5/3)], where r(0) is Fried's coherence length and d is the average interval of the actuators on the deformable mirror. The fitting parameter kappa is 0.45. Numerical simulations are also performed with the experimental parameters. The numerical results are in agreement with data obtained in the experiment, which shows that the direct-tilt phase-reconstruction method used in our four dimensional simulation code is reasonable. PMID- 18285910 TI - Horizontal Line-of-Sight Turbulence Over Near-Ground Paths and Implications for Adaptive Optics Corrections in Laser Communications. AB - Atmospheric turbulence over long horizontal paths perturbs phase and can also cause severe intensity scintillation in the pupil of an optical communications receiver, which limits the data rate over which intensity-based modulation schemes can operate. The feasibility of using low-order adaptive optics by applying phase-only corrections over horizontal propagation paths is investigated. A Shack-Hartmann wave-front sensor was built and data were gathered on paths 1 m above ground and between a 1- and 2.5-km range. Both intensity fluctuations and optical path fluctuation statistics were gathered within a single frame, and the wave-front reconstructor was modified to allow for scintillated data. The temporal power spectral density for various Zernike polynomial modes was used to determine the effects of the expected corrections by adaptive optics. The slopes of the inertial subrange of turbulence were found to be less than predicted by Kolmogorov theory with an infinite outer scale, and the distribution of variance explained by increasing order was also found to be different. Statistical analysis of these data in the 1-km range indicates that at communications wavelengths of 1.3 mum, a significant improvement in transmitted beam quality could be expected most of the time, to a performance of 10% Strehl ratio or better. PMID- 18285911 TI - Partial correction for turbulent distortions in telescopes. AB - To provide complete compensation for turbulent distortions in the visible range at aperture dimensions typical for modern telescopes (6-10 m), one needs to develop adaptive systems with hundreds of control channels. More simple adaptive systems that provide complete compensation in the infrared range can give an essential advantage in angular resolution in the visible range too. In this case the image brightness characterized by the Strehl ratio remains much less than that in the diffraction-limited case, i.e., the system provides only partial compensation. We present the results of numerical calculations of the partially corrected point-spread function and discuss possible approaches to composing the adaptive system configuration. PMID- 18285912 TI - Spectrometer slit-power-coupling calculations for natural and laser guide-star adaptive optics. AB - We use numerical calculations to examine the relation between adaptive optics (AO) turbulence compensation and power coupled through a spectrometer slit for both laser and natural guide-star AO systems. The AO system and observing parameters used are relevant to the Gemini-North 8-m telescope. For this study, we separate residual tilt from residual higher-order aberrations to isolate their relative effects under a variety of operating conditions. Our results demonstrate that slit-coupled intensity is not uniquely determined by system Strehl alone; we show that this is due to the differing effects of higher-order and tilt aberrations on the shape of the compensated point-spread function. For the Gemini spectrometer and AO system, the wider point-spread function halo associated with an added residual higher-order aberration reduces slit-coupled intensity more rapidly than a broad point-spread function core induced by residual tilt. PMID- 18285913 TI - Two-deformable-mirror concept for correcting scintillation effects in laser beam projection through the turbulent atmosphere. AB - A two-deformable-mirror concept for correcting scintillation effects in laser beam projection through the turbulent atmosphere is presented. This system uses a deformable mirror and a Fourier-transforming mirror to adjust the amplitude of the wave front in the telescope pupil, similar to kinoforms used in laser beam shaping. A second deformable mirror is used to correct the phase of the wave front before it leaves the aperture. The phase applied to the deformable mirror used for controlling the beam amplitude is obtained with a technique based on the Fienup phase-retrieval algorithm. Simulations of propagation through a single turbulent layer sufficiently distant from the beacon observation and laser beam transmission aperture to cause scintillation shows that, for an ideal deformable mirror system, this field-conjugation approach improves the on-axis field amplitude by a factor of approximately 1.4 to 1.5 compared with a conventional phase-only correction system. PMID- 18285914 TI - Real-time modal control implementation for adaptive optics. AB - The electronics, computing hardware, and computing used to provide real-time modal control for a laser guide-star adaptive optics system are presented. This approach offers advantages in the control of unobserved modes, the elimination of unwanted modes (e.g., tip and tilt), and automatic handling of the case of low resolution lens arrays. In our two-step modal implementation, the input vector of gradients is first decomposed into a Zernike polynomial mode by a least-squares estimate. The number of modes is assumed to be less than or equal to the number of actuators. The mode coefficients are then available for collection and analysis or for the application of modal weights. Thus the modal weights may be changed quickly without recalculating the full matrix. The control-loop integrators are at this point in the algorithm. To calculate the deformable mirror drive signals, the mode coefficients are converted to the zonal signals by a matrix multiply. When the number of gradients measured is less than the number of actuators, the integration in the control loop will be done on the lower resolution grid to avoid growth of unobserved modes. These low-resolution data will then be effectively interpolated to yield the deformable-mirror drive signals. PMID- 18285915 TI - Analysis of algorithms for adaptive control of laser beams in a nonlinear medium. AB - Adaptive correction for thermal blooming by multidither and phase-conjugation algorithms is analyzed with the use of numerical models of beams propagating in a nonlinear medium. Methods to increase the stability of these algorithms are proposed. The influence of an adaptive mirror on the efficiency and the divergence of correction algorithms is considered. The feasibility of amplitude phase control with a two-mirror adaptive optics system is also analyzed. PMID- 18285916 TI - Performance analysis of and compensation for aspect-ratio effects of fast-fourier transform-based simulations of large atmospheric wave fronts. AB - Fast-Fourier-transform-based simulators of atmospheric wave fronts with a von Karman turbulence spectrum were tested with reference to the phase-structure function and phase variance over a pupil on large square and rectangular formats. The symmetry and the accuracy of the phase-structure function were found to be limited by the aspect ratio and the size of the phase screen. The phase variance over a pupil is less sensitive to the aspect ratio than the phase-structure function and is dependent mainly on the size of the phase screen. Several tests are reported and discussed together with a method of compensation for the negative effects of rectangular formats. PMID- 18285917 TI - Myopic deconvolution of adaptive optics images by use of object and point-spread function power spectra. AB - Adaptive optics systems provide a real-time compensation for atmospheric turbulence. However, the correction is often only partial, and a deconvolution is required for reaching the diffraction limit. The need for a regularized deconvolution is discussed, and such a deconvolution technique is presented. This technique incorporates a positivity constraint and some a priori knowledge of the object (an estimate of its local mean and a model for its power spectral density). This method is then extended to the case of an unknown point-spread function, still taking advantage of similar a priori information on the point spread function. Deconvolution results are presented for both simulated and experimental data. PMID- 18285918 TI - Optimization of a predictive controller for closed-loop adaptive optics. AB - For closed-loop adaptive optics systems limited by time delay and measurement noise, we demonstrate that the ideal rejection transfer function is proportional to the frequency signal-to-noise ratio of the wave-front input. We describe a new modal linear predictive controller that approaches this ideal transfer function. Its parameters are optimized by minimization of the residual wave-front error with a modified recursive least-squares algorithm. The optimization can be performed with closed-loop data in the case of evolving turbulent conditions. We present numerical simulations to show the significant improvements brought by the predictor. PMID- 18285919 TI - Monostatic and bistatic schemes and an optimal algorithm for tilt correction in ground-based adaptive telescopes. AB - It has been pointed out that the two terms effective scattering volume and laser guide star are scientific synonyms. The first term was introduced by the end of 1970 by specialists in atmospheric optics and laser sounding. Information has also been given concerning fluctuations of waves reflected from an object and waves passed twice through atmospheric inhomogeneities. A corresponding mathematical apparatus allows one to estimate correlation characteristics of a laser guide star. The mutual correlation function of random angular displacements of a plane-wave image and the centroid displacements of a focused Gaussian laser beam are considered. The algorithm of optimal correction is introduced in which a priori information is used. Based on this algorithm, the variance of residual jitter of a star image obtained with the use of a laser guide star is evaluated. PMID- 18285920 TI - Minimum diameter of a laser projector for some perspective-based laser guide-star tilt retrieval schemes. AB - Because of the finite speed of light, a laser guide Star that is seen from the side is not exactly a straight line. When such a laser guide star is used to sense tip-tilt with some of the perspective-based techniques that are used to retrieve an absolute tip-tilt laser, such nonstraightness introduces an error. We estimate this effect for various diameters of a laser projector, assuming a Kolmogorov turbulence spectrum and figuring the maximum achievable Strehl ratio. We found that under poor seeing conditions the effect is not negligible and that laser projectors larger than those currently used are required. PMID- 18285921 TI - Membrane mirror and bias electronics. AB - We have designed and built an electrostatically deformable membrane mirror with simple bias and driver electronics to evaluate its suitability for a curvature sensing adaptive optics system. It has a 100-mm-diameter aluminized nitrocellulose membrane, with 31 actuators arranged concentrically. The unit operates at atmospheric pressure with a high bias voltage applied to the membrane. The high-voltage electronics are contained within the mirror housing for safety reasons. An entrance window reduces the effects of air-coupled vibration. Details of the device and design rationale are presented. With a proper bias, the unit can provide low-order (including tip-tilt) wave-front correction. PMID- 18285922 TI - Design of an adaptive secondary mirror: a global approach. AB - We present the mechanical and actuator design of an adaptive secondary mirror that matches the optical requirements of the active and adaptive corrections. Conceived for the particular implementation for the 6.5-m conversion of the multiple-mirror telescope, with small variations of the input parameters this study is suitable for applications for telescopes of the same class. We found that a three-layer structure, i.e., a thin deformable shell, a thick reference plate, and a third plate that acts as actuator support and heat sink, is able to provide the required mechanical stability and actuator density. We also found that a simple electromagnetic actuator can be used. This actuator, when optimized, will dissipate a typical power of a few tenths of watts. PMID- 18285923 TI - Low-order adaptive deformable mirror. AB - We report the main parameters of a nine-electrode bimorph piezoelectric adaptive mirror designed to correct low-order aberrations. We describe measurements of the control coefficients for defocus, astigmatism, pure coma, and spherical aberration of this mirror and the temperature stability of its profile. The performance of a simple adaptive optical system for imaging through laboratory generated turbulence is investigated. This low-order device is suitable for small (<1-m-diameter) telescopes and for nonastronomical applications of adaptive optics. PMID- 18285924 TI - Comparison of near-forward light scattering on oceanic turbulence and particles. AB - We examine and compare near-forward light scattering that is caused by turbulence and typical particulate assemblages in the ocean. The near-forward scattering by particles was calculated using Mie theory for homogeneous spheres and particle size distributions representative of natural assemblages in the ocean. Direct numerical simulations of a passive scalar with Prandtl number 7 mixed by homogeneous turbulence were used to represent temperature fluctuations and resulting inhomogeneities in the refractive index of water. Light scattering on the simulated turbulent flow was calculated using the geometrical-optics approximation. We found that the smallest temperature scales contribute the most to scattering, and that scattering on turbulence typically dominates over scattering on particles for small angles as large as 0.1 degrees . The scattering angle deviation that is due to turbulence for a light beam propagating over a 0.25-m path length in the oceanic water can be as large as 0.1 degrees . In addition, we carried out a preliminary laboratory experiment that illustrates the differences in the near-forward scattering on refractive-index inhomogeneities and particles. PMID- 18285925 TI - Determination of the Atmospheric-Water-Vapor Content in the 940-nm Absorption Band by Use of Moderate Spectral-Resolution Measurements of Direct Solar Irradiance. AB - We have analyzed three methods that can be used to determine the integrated water vapor of the atmosphere in the 940-nm band by means of modeled and measured direct solar spectral irradiance. The experimental irradiance data were obtained with a commercial LI-COR 1800 spectroradiometer, based on a monochromator system, of high to moderate spectral resolution (6 nm) in the 300-1100-nm range. The modeled data are based on monochromatic approaches to determine atmospheric transmittance constituents; for those of water vapor we used the lowtran7 model. The first method is a curve-fitting procedure that makes use of the entire shape band absorption information to retrieve a unique water-vapor value. The second method makes use of the monochromatic approach of the absorption transmittance formula to determine the amount of water vapor at each wavelength of the absorption band, and the third method is the classic differential absorption technique suitably applied to our data. Spectral analysis showed the advantages and disadvantages of each method, such as problems linked to the various spectral resolutions of the experimental and the modeled data, the width of the spectral range used to define the water-vapor absorption band, and the dependence of the retrieval on the choice of the two selected wavelengths in the last-named technique. All these problems were considered so they could be avoided or minimized and the associated errors estimated. We used the methods to determine water-vapor values for the period from March to November 1995 at a rural station in Vallodolid, Spain, allowing for the evaluation of the differences in real monitoring conditions. Finally, the contribution of continuum absorption was also evaluated, yielding lower water-vapor values between 13 and 30%. These differences were considerably greater than those that were due to the problems that we have just enumerated. PMID- 18285926 TI - Iterative method for the inversion of multiwavelength lidar signals to determine aerosol size distribution. AB - Two iterative methods of inverting lidar backscatter signals to determine altitude profiles of aerosol extinction and altitude-resolved aerosol size distribution (ASD) are presented. The first method is for inverting two wavelength lidar signals in which the shape of the ASD is assumed to be of power law type, and the second method is for inverting multiwavelength lidar signals without assuming any a priori analytical form of ASD. An arbitrary value of the aerosol extinction-to-backscatter ratio (S(1)) is assumed initially to invert the lidar signals, and the ASD determined by use of the spectral dependence of the retrieved aerosol extinction coefficients is used to improve the value of S(1) iteratively. The methods are tested for different forms of altitude-dependent ASD's by use of simulated lidar-backscatter-signal profiles. The effect of random noise on the lidar backscatter signals is also studied. PMID- 18285927 TI - Beam spread function with time dispersion. AB - An analytical model for the beam spread function with time dispersion is modeled and validated against Monte Carlo calculations. The model, which is structured on simple statistical concepts and relies on only first and second moments for displacement, angle, and multipath time, is suitable for describing pulsed laser radiation propagation in nonconservative scattering media out to tens of scattering lengths. Numerical examples for marine environments are used to show its robustness and versatility. PMID- 18285928 TI - Parameterization of Stratospheric Aerosol Physical Properties on the Basis of Nd:YAG Lidar Observations. AB - An extension to the 355- and 1064-nm wavelengths of a numerical optical model originally developed at 532 nm is presented. The resulting parameterization allows estimates of stratospheric aerosol surface area, volume, and extinction-to backscatter ratio from lidar measurements obtained at one of the two Nd:YAG laser wavelengths. Functional relationships that link single-wavelength backscatter to each of the physical variables are provided for sulfate aerosol types ranging from background to heavy volcanic under environmental conditions representative of the global lower stratosphere. The behavior of the functional relationships at the three Nd:YAG wavelengths is compared. Relative errors of model estimates range between 10% and 50%, depending on wavelength and backscatter cross sections. These values are comparable with the ones that characterize in situ particle counters. The inference of particle effective radius and the application of the method to the interpretation of supercooled polar stratospheric cloud observations are discussed. PMID- 18285929 TI - Global tropospheric and total ozone monitoring with a double-etalon fabry-perot interferometer. I. Instrument concept. AB - The global distribution of tropospheric ozone (O(3)) can be observed from a satellite-based instrument by spectrally isolating the pressure-broadened wings of strong O(3) lines. The Fabry-Perot interferometer (FPI) provides high spectral resolution and high-throughput capabilities that are essential for performing such a measurement. Through proper selection of channel spectral regions, the FPI optimized for tropospheric O(3) measurements can simultaneously observe a stratospheric component and thus the total O(3) column abundance. We present a conceptual instrument design that involves a double-etalon fixed-gap series configuration FPI along with an ultranarrow bandpass filter to achieve single order operation with an overall spectral resolution of approximately 0.068 cm( 1), sampling the narrow 1054.2-1055.2 cm(-1) spectral region within the strong 9.6-mum ozone infrared band from a nadir-viewing satellite configuration. PMID- 18285930 TI - Global Tropospheric and Total Ozone Monitoring with a Double-Etalon Fabry-Perot Interferometer. II. Feasibility Analysis. AB - The feasibility of observing global tropospheric and total ozone (O(3)) fields with a double-etalon Fabry-Perot interferometer (FPI) has been assessed. The FPI provides high spectral resolution and high-throughput capabilities that enable observation in pressure-broadened wings of strong O(3) lines while minimizing the impact of undesirable signal contributions (i.e., from the terrestrial surface and interfering species). A retrieval technique has been implemented and is demonstrated for a tropical atmosphere possessing enhanced tropospheric ozone amounts. An error analysis assessing the impact on retrieved O(3) amounts from the most significant uncertainties associated with this particular measurement has been performed, and findings for a tropical atmosphere are presented. Results show the proposed instrumentation to enable a good measurement of absolute ozone amounts and an even better determination of relative changes. PMID- 18285931 TI - Spatial variability of oceanic phycoerythrin spectral types derived from airborne laser-induced fluorescence emissions. AB - We report spatial variability of oceanic phycoerythrin spectral types detected by means of a blue spectral shift in airborne laser-induced fluorescence emission. The blue shift of the phycoerythrobilin fluorescence is known from laboratory studies to be induced by phycourobilin chromophore substitution at phycoerythrobilin chromophore sites in some strains of phycoerythrin-containing marine cyanobacteria. The airborne 532-nm laser-induced phycoerythrin fluorescence of the upper oceanic volume showed distinct segregation of cyanobacterial chromophore types in a flight transect from coastal water to the Sargasso Sea in the western North Atlantic. High phycourobilin levels were restricted to the oceanic (oligotrophic) end of the flight transect, in agreement with historical ship findings. These remotely observed phycoerythrin spectral fluorescence shifts have the potential to permit rapid, wide-area studies of the spatial variability of spectrally distinct cyanobacteria, especially across interfacial regions of coastal and oceanic water masses. Airborne laser-induced phytoplankton spectral fluorescence observations also further the development of satellite algorithms for passive detection of phytoplankton pigments. Optical modifications to the NASA Airborne Oceanographic Lidar are briefly described that permitted observation of the fluorescence spectral shifts. PMID- 18285932 TI - Physically based parameterizations of the short-wave radiative characteristics of weakly absorbing optically thick media: application to liquid-water clouds. AB - We propose the physically based parameterization of the radiative characteristics of liquid-water clouds as functions of the wavelength, effective radius, and refractive index of particles, liquid-water path, ground albedo, and solar and observation angles. The formulas obtained are based on the approximate analytical solutions of the radiative transfer equation for optically thick, weakly absorbing layers and the geometrical optics approximation for local optical characteristics of cloud media. The accuracy of the approximate formulas was studied with an exact radiative transfer code. The relative error of the approximate formula for the reflection function at nadir observations was less then 15% for an optical thickness larger than 10 and a single-scattering albedo larger than 0.95. PMID- 18285933 TI - Self-aligning lidar for the continuous monitoring of the atmosphere. AB - In a lidar system an accurate transmitter-receiver alignment is essential for correct results. Usually this optical adjustment is time consuming and requires the intervention of highly qualified personnel. As a solution to this problem, a fast and precise automatic alignment procedure is presented, based on a simple model of the transmitter-receiver overlap. The lidar mounted at the Naples University is used to test this method. A centering precision of few microradians is obtained through dedicated software controlling a gimbal-mounted mirror. The automatic alignment procedure is then assessed. In particular, the correctness of the center and of its error is determined. Finally, the system is applied to the monitoring of tropospheric aerosols, leading to the continuous retrieval of profiles with fine spatiotemporal resolution. PMID- 18285934 TI - Apparent optical properties of oceanic water: dependence on the molecular scattering contribution. AB - The relationships between the apparent optical properties (AOP's) and the inherent optical properties (IOP's) of oceanic water bodies have been reinvestigated by solution of the radiative transfer equation. This reexamination deals specifically with oceanic case 1 waters (those for which phytoplankton and their associated particles or substances control their inherent optical properties). In such waters, when the chlorophyll content is low enough (in most of the entire ocean), the influence of molecular scattering by water molecules is not negligible, leading to a gradual change in the shape of the phase function. The effect of this change on the AOP's is analyzed. The effect of the existence of diffuse sky radiation in addition to the direct solar radiation on AOP-IOP relationships is also examined. Practical parameterizations are proposed to predict in case 1 waters, and at various depths, the vertical attenuation coefficient for downward irradiance (K(d)) as a function of the IOP's and solar angle. These parameterizations are valid for the spectral domain where inelastic scattering does not significantly occur (wavelengths below 590 nm). PMID- 18285935 TI - Single-Crystal Laser-Heated Pedestal-Growth Sapphire Fibers for Er:YAG Laser Power Delivery. AB - The Er:YAG laser-induced damage (LID) threshold and modal properties of single crystal sapphire fibers grown by the laser-heated pedestal-growth method have been measured. The lowest loss (~0.4-dB/m) sapphire fibers produce little mode mixing and therefore deliver a near-single-mode output profile if the Er:YAG laser input beam profile is also nearly Gaussian. Normally, however, Er:YAG laser output beam profiles are multimode with numerous high-energy spikes. This leads not only to a multimode output from the fiber but also increased fiber loss that is due to higher-order mode coupling. The results of LID testing give a damage fluence of ~1.4 kJ/cm(2) for 300-mum core-only sapphire fibers at 2.94 mum. PMID- 18285936 TI - Experimental verification and theory for an eight-element multiple-aperture equal gain coherent laser receiver for laser communications. AB - The detection and processing of laser communication signals are affected by the fading induced onto these signals by atmospheric turbulence. One method of reducing this fading is to use an array of detectors in which each of the detector outputs are added together coherently. We present experimental verification and theory of a 1.06 mum eight-element coherent receiver used to mitigate the effects of fading over a 1-km outdoor range. The carrier-to-noise ratio (CNR) was measured on a single channel and was then compared with the CNR obtained from the coherent sum of the eight channels. The increase of the mean CNR for the coherent sum as compared with a single aperture was observed proportional to the number of the apertures under different conditions of atmospheric turbulence. The measured mean CNR gain fitted the theoretical prediction well when the laser intensity fluctuations followed the gamma distribution. PMID- 18285937 TI - Laser-to-fiber optical coupling scheme with a long working distance by use of thermally overexpanded fiber. AB - A laser-diode-optical-fiber coupling scheme that uses aspherically ended thermally overexpanded fiber is proposed. The scheme is verified by both an analytical formalism and a wide-angle beam-propagation method analysis in cylindrical coordinates. PMID- 18285938 TI - Linearly polarized er(3+)-doped fiber laser by use of a fiber loop polarizer and a wavelength-selective fiber loop reflector. AB - We demonstrate a novel single-polarization Er(3+)-doped fiber Fabry-Perot laser. The all-fiber Fabry-Perot cavity is simple inasmuch as it is formed by only two fiber-based components, a fiber loop polarizer and a wavelength-selective fiber loop reflector. The threshold power is 6.5 mW, the slope efficiency is 12.2%, and the extinction ratio is approximately 25 dB. PMID- 18285939 TI - Nonlinear holographic imaging of phase errors. AB - Experimental measurements and computer simulations of nonlinear holographic imaging of phase errors in laser beams are presented. The computer models are found to accurately predict the results of the experiments. Comparison with similar results by use of amplitude scatterers reveals that the image location (along the propagation path) is the same for phase and amplitude scatterers. However, the intensity and fluence of the image of a phase scatterer are significantly larger, indicating that phase objects pose a larger damage threat to optical components. PMID- 18285940 TI - Application of zeeman modulation faraday spectroscopy to the measurement of a magnetic field. AB - We report what is to our knowledge the first possibility of a NO(2) molecular magnetometer based on the Zeeman modulation magnetic rotation spectroscopic (ZM MRS) technique and the magneto-optic activity of NO(2). The linear dependence of the ZM MRS signal intensity on the modulating magnetic field is theoretically analyzed and experimentally measured. The design concept of the magnetometer and its main features are discussed. PMID- 18285941 TI - Development of a System for Studying the Morphology and Dynamics of Agglomerated Flame Particulates by use of Dynamic Light Scattering. AB - An experimental system for the study of the morphological and dynamic properties of flame-generated agglomerates by use of both polarized (vertical-vertical polarization orientation) and depolarized (vertical-horizontal polarization orientation) dynamic light scattering (DLS) was developed and tested. The system consists of a flame reactor for generating chainlike agglomerates of Fe(2)O(3) in an Fe(CO)(5)-seeded CO-O(2) diffusion flame and a light-scattering spectrometer for performing polarized and depolarized DLS measurements of the agglomerates' dynamic properties. It is demonstrated for the first time that one can successfully obtain depolarized DLS correlation functions from a flame environment by combining the results of a series of measurements obtained using a cross-correlation detection system. PMID- 18285942 TI - Throughput of tilted interferometers. AB - The throughput of a tilted Fourier-transform spectrometer (FTS) with collimation is calculated. It is shown that the maximum off-axis angle that is acceptable in the interferometer is inversely proportional to the distance between the detector and the location where the tilt is applied to the wave fronts and is also inversely proportional to the tilt angle. This effect leads to tilt sensitivity in a scanning FTS and to the loss of the throughput advantage in a FTS with no moving part in which a tilt between two collimated beams is used to disperse the interferogram spatially. Experimental verification confirms the throughput condition with tilt angle. PMID- 18285943 TI - Comparisons of laser-saturated, laser-induced, and planar laser-induced fluorescence measurements of nitric oxide in a lean direct-injection spray flame. AB - We report quantitative, spatially resolved laser-saturated fluorescence (LSF), linear laser-induced fluorescence (LIF), and planar laser-induced fluorescence (PLIF) measurements of nitric oxide (NO) concentration in a preheated, lean direct-injection spray flame at atmospheric pressure. The spray is produced by a hollow-cone, pressure-atomized nozzle supplied with liquid heptane, and the overall equivalence ratio is unity. NO is excited by means of the Q(2)(26.5) transition of the gamma(0, 0) band. LSF and LIF detection are performed in a 2-nm region centered on the gamma(0, 1) band. PLIF detection is performed in a broad ~70-nm region with a peak transmission at 270 nm. Quantitative radial NO profiles obtained by LSF are presented and analyzed so as to correct similar LIF and PLIF profiles. Excellent agreement is achieved among the three fluorescence methodologies. PMID- 18285944 TI - Noninvasive magnetometry based on magnetic rotation spectroscopy of oxygen. AB - An experimental demonstration of a noninvasive optical probe of magnetic fields is presented. The technique used is magnetic rotation spectroscopy of the b(1)?(g)(+) - X(3)?(g)(-) band of oxygen near 760 nm. Ambient concentrations of oxygen at atmospheric pressure are sufficient to observe substantial signals. In addition, a diode laser is used as the light source, making this a simple and compact measurement possibility. PMID- 18285945 TI - Beam propagation (m(2)) measurement made as easy as it gets: the four-cuts method. AB - Tolerance analysis shows that an efficient M(2) measurement plan is a first cut (beam diameter measurement) at 0.5-2.0 Rayleigh ranges to one side of the waist, which is matched by interpolation between second and third cuts to the opposite side. The waist is measured by a fourth cut halfway between the matched diameters, yielding an easy two-parameter curve fit for M(2). PMID- 18285946 TI - Ray analysis of parabolic-index segmented planar waveguides. AB - A ray analysis of periodically segmented waveguides with parabolic-index variation in the high-index region is presented. We carried out the analysis using ray transfer matrices, which is convenient to implement and which can be extended to study different types of graded-index segmented waveguide. Results of this ray tracing approach clearly illustrate the waveguiding properties and the existence of stable and unstable regions of operation in segmented waveguides. We also illustrate the tapering action exhibited by segmented waveguides in which the duty cycle varies along the length of the waveguide. This analysis, although restricted to multimode structures, provides a clear visualization of the waveguiding properties in terms of ray propagation in segmented waveguides. PMID- 18285947 TI - Diode-Pumped Emission of Tm(3)(+)-Doped Ca(2)Al(2)SiO(7) Crystals. AB - A diode-pumped Tm:Ca(2)Al(2)SiO(7) (Tm:CAS) laser has been demonstrated for the first time to the authors' knowledge. A 39-mW output power and an 8.6% slope efficiency were obtained at -11 degrees C. The most attractive features of Tm:CAS are a broad absorption band near 785 nm and a large ground-state splitting. The improvement in laser performance expected from the large Stark splitting is shown to be limited by the enhancement of multiphonon relaxation processes and by the low thermal conductivity of the crystal. PMID- 18285948 TI - Standardization of the definition and measurement of the line length and fan angle of laser line generators. AB - Three different commonly used methods of measuring the length and fan angle of a light line or a light fan produced by a laser line generator were analyzed, tested, and compared. One measurement method is proposed as the standard method, and the resultant 1/e(2) intensity fan angle is proposed as the standard parameter to describe the line generator. PMID- 18285949 TI - Suppression of amplified spontaneous emission from a four-pass dye laser amplifier. AB - We demonstrate a new design for a four-pass dye laser amplifier that can be used to reduce amplified spontaneous emission (ASE) in the amplifier output beam. Compared with the results obtained from normal operation of the four-pass dye laser amplifier without a frequency-selective device, by using a diffraction grating in the amplifier we can reduce the ASE ratio by a factor in excess of 10 and increase output energy by ~4%. The obtained ASE ratio of the four-pass amplifier system is <1.5%. PMID- 18285950 TI - Narrow-linewidth master-oscillator power amplifier based on a semiconductor tapered amplifier. AB - The output of a grating-stabilized external-cavity diode laser was injected into a semiconductor tapered amplifier in a master-oscillator power amplifier configuration, producing as much as 500 mW of power with narrow linewidth. The additional linewidth that is due to the tapered amplifier is much smaller than the typical linewidth of grating-stabilized laser diodes. To demonstrate the usefulness of the narrow linewidth and high output power, we used the system to perform Doppler-free two-photon spectroscopy with rubidium. PMID- 18285951 TI - Tunable picosecond blue and ultraviolet pulses from a diode-pumped laser system seeded by a gain-switched laser diode. AB - Picosecond pulses emitted from a gain-switched laser diode have been amplified in a Ti:sapphire regenerative amplifier indirectly pumped by a 4-W laser diode. This all-solid-state system produced microjoule pulses tunable from 803 to 840 nm at repetition rates up to 25 kHz with durations of 70-100 ps. By frequency doubling and tripling the output, we generated blue and UV pulses tunable from 401 to 420 nm and from 268 to 280 nm, respectively. Average powers larger than 4 mW were reached in these two wavelength regions. PMID- 18285952 TI - Output Power Optimization and Gain and Saturation Irradiance Measurements on a RF Pumped HCN Waveguide Laser. AB - A 1-m-long RF-pumped HCN waveguide laser is described. Output power optimizations were performed with mixtures of methane, nitrogen, and helium in the ratios 1:2:0, 1:1:0, 2:1:0, and 8:3:36. The variation of output power as a function of gas pressure, flow rate, waveguide wall temperature, and RF driving power was studied. A maximum output power of approximately 50 mW was obtained from the 337 mum line in the EH(11) cavity mode with the 1:1:0 mixture. Gain and saturation irradiance measurements were also performed on the same laser with an adjustable Michelson output coupler. The output power was measured as a function of the adjustment of the coupler, and a least-squares fit of a suitable laser model to the measured data was employed to furnish values for the small signal gain and saturation irradiance of the gain medium. Gain and saturation irradiance values were measured in this way for the above gas mixtures as a function of gas pressure, flow rate, and wall temperature, and the results were used to calculate the corresponding output powers. Reasonable agreement was found between these calculated powers and experimentally measured output powers. A quadratic variation of saturation irradiance with pressure is predicted by a simple theory and is also observed in the measured data. PMID- 18285953 TI - Continuous-Wave, All-Solid-State, Single-Frequency 400-mW Source at 589 nm Based on Doubly Resonant Sum-Frequency Mixing in a Monolithic Lithium Niobate Resonator. AB - Sum-frequency mixing of two cw single-mode Nd:YAG lasers in a doubly resonant congruent lithium niobate resonator generated two TEM(00) beams of single frequency 589-nm radiation. The primary beam had a power of 400 mW and the secondary beam of approximately 15 mW by use of 320 mW of 1319-nm and 660 mW of 1064-nm Nd:YAG radiation incident on the lithium niobate resonator. This corresponds to an optical power conversion efficiency of more than 40%. PMID- 18285954 TI - Technique for continuous tuning of optical fiber lasers. AB - For the first time to the authors' knowledge, we have demonstrated how thermally controlled overcoupled fused fiber couplers and fiber loop mirrors based on these couplers can be used as broadband tuning elements in a fiber laser cavity. No bulk optical elements play any role in this technique. Temperature tuning the coupler results in a shift in the coupling ratio or in the effective output coupler transmission. For a fixed pump source, and for a given laser cavity, this shift causes the lasing wavelength to shift. We have continuously tuned an Er silica fiber laser in this manner over the range of 1527-1570 nm in a ring configuration, and, using a fiber loop mirror with these couplers in a linear Tm silica fiber laser cavity, we have achieved more than 50-nm broadband tuning over the range of 1850-1910 nm. The tuning range and the sensitivity to temperature depend on the degree of overcoupling of the loop mirror coupler. PMID- 18285955 TI - Experimental determination of the fundamental-mode diameter in solid-state lasers. AB - A method to determine directly the radius of the fundamental mode in a laser crystal has been developed. The radius is measured by comparison of the distribution of the spontaneous emission in the pumped region during laser operation with the fluorescence distribution without laser emission. Measuring the mode radius with various pump powers enables one to optimize the overlap between the pump and the cavity beam and to determine the dioptric power of the thermally induced lens. PMID- 18285956 TI - Pulse amplification of a single-frequency cr:forsterite laser. AB - Stable single-longitudinal-mode operation of a tunable gain-switched Cr:forsterite oscillator-amplifier system is reported. A novel coupled-cavity oscillator configuration provides a low-threshold fluence of 0.3 J cm(-2), making the system attractive for laser media with low gain or high parasitic loss. Nearly transform-limited pulses with instrument-limited bandwidths of 150 MHz have been obtained across a 100-nm wavelength range, limited by the cavity optics. A range of forsterite crystals with Cr(4+) concentration in the 3-21 x 10(18)-cm(-3) range has been used for investigation of amplifier performance in single- and double-pass configurations. PMID- 18285957 TI - Spectroscopy and laser characteristics of copper-vapor-laser pumped pyrromethene 556 and pyrromethene-567 dye solutions. AB - We measured the basic optical properties of Pyrromethene-567 (P567) and Pyrromethene-556 (P556) dye solutions that are relevant to their application as dye lasers. The fluorescence spectra of methanol solutions show mirror images in relation to the absorption spectra, with Stokes shifts of 29.5 and 37.5 nm, respectively, for the two dyes. The central fluorescence peaks were at 546 and 535 nm, with widths of ~40 and ~50 nm (FWHM). The quantum yields were 97% ? 5% and 78% ? 5% for P567 and P556, respectively. Fluorescence lifetimes of 6.0 ? 0.2 ns were obtained for both dyes in methanol. Laser action, obtained by pumping with the green emission line (510.6 nm) from a copper-vapor laser, was measured in a Hansch-type cavity. Tunability ranged from 531 to 590 nm for P567 and from 522 to 590 nm for P556. Lasing thresholds were ~0.27 and ~0.16 mJ/pulse, with 25% and 27% slope efficiencies for P567 and P556, respectively. Spectroscopy and lasing were studied in other solvents as well. PMID- 18285958 TI - Utilization of the Yellow Component of a Copper-Vapor Laser for Extending the Tuning Range of a Rhodamine 6G Dye Laser by Use of an Additional Dye in a Novel Coupled Resonator Scheme. AB - We demonstrate the use of both 510.6- and 578.2-nm components and the extension of the tuning range of a Rhodamine 6G dye laser in a novel coupled resonator scheme. Rhodamine 6G is pumped by 510.6-nm light in one resonator and Sulforhodamine B is pumped by 578.2 nm in the other. The spectral tuning range of 564-609 nm of the Rhodamine 6G laser is extended up to 640 nm. A two-mirror arrangement ensures continuous tuning across the spectral ranges of the two dyes by rotation of a single plane mirror. PMID- 18285959 TI - Widely Tunable Continuous-Wave Mid-Infrared Laser Source Based on Difference Frequency Generation in AgGaS(2). AB - We demonstrate difference-frequency generation in the 6.8-12.5-mum range by mixing two high-power single-frequency laser diodes in a type II AgGaS(2) crystal. This compact all-solid-state scheme provides maximum output powers that exceed 1 muW and permits continuous adjustment-free scans larger than 2 cm(-1) across the entire tuning range. PMID- 18285960 TI - Widely Tunable Continuous-Wave CR(3+):LiSrAlF(6) Ring Laser from 800 to 936 nm. AB - We report on a widely tunable, diode-pumped, continuous-wave, single-frequency Cr(3+):LiSrAlF(6) laser with a novel double-ring cavity configuration. The optical diode and the tuning element were located in the ring subcavity to decrease their insertion loss. Using an 1800-lines/mm grating as the tuning element, we obtained a laser emission of 20-mW output power at 870 nm and a tuning range of from 800 to 936 nm. When we used a 1200-lines/mm grating with a higher diffraction efficiency, the laser had a slightly wider tuning range of from 800 to 938 nm. However, the output power decreased at this tuning range. PMID- 18285961 TI - Refractive-Index Measurements of Undoped Yttrium Aluminum Garnet from 0.4 to 5.0 mum. AB - We report measurements of the refractive index of undoped yttrium aluminum garnet from 0.4 to 5.0 mum and the calculation of Sellmeier coefficients based on our data. The data differ considerably from previously published reports. The effect of the new data on the design of optical devices such as intracavity etalons for lasers is discussed. PMID- 18285962 TI - Response of a Fabry-Perot Optical Cavity to Phase Modulation Sidebands for use in Electro-Optic Control Systems: Errata. PMID- 18285963 TI - One-Dimensional, Time-Resolved Raman Measurements in a Sooting Flame made with 355-nm Excitation. AB - Single-shot vibrational Raman measurements were performed along an 11-mm-long line crossing the reaction zone in a premixed, fuel-rich (phi = 10), laminar methane-air flame by use of a frequency-tripled Nd:YAG laser with a 355-nm emission wavelength. This laser source seems to have advantages relative to KrF excimer lasers as well as to Nd:YAG lasers at 532 nm for hydrocarbon combustion diagnostics. The Raman emissions of all major species (N(2), O(2), CH(4), H(2), CO(2), H(2)O) were detected simultaneously with a spatial resolution of 0.4 mm. By integration over selected spectral intervals, the mole fractions of all species and subsequently the local gas temperatures have been obtained. A comparison of the temperatures that were found with results from filtered Rayleigh experiments showed good agreement, indicating the success of what are to the best of our knowledge the first one-dimensional single-shot Raman measurements in a sooting hydrocarbon flame. PMID- 18285964 TI - Remote-sensing technique for determination of the volume absorption coefficient of turbid water. AB - We use remote-sensing reflectance from particulate R(rs) to determine the volume absorption coefficient a of turbid water in the 400 < lambda < 700-nm spectral region. The calculated and measured values of a(lambda) show good agreement for 0.5 < a < 10 (m(-1)). To determine R(rs) from a particulate, we needed to make corrections for remote-sensing reflectance owing to surface roughness S(rs). We determined the average spectral distribution of S(rs) from the difference in total remote-sensing reflectance measured with and without polarization. The spectral shape of S(rs) showed an excellent fit to theoretical formulas for glare based on Rayleigh and aerosol scattering from the atmosphere. PMID- 18285965 TI - Disturbance of laser-induced-fluorescence measurements of NO in methane-air flames containing chlorinated species by photochemical effects induced by 225-nm laser excitation. AB - Laser-induced fluorescence measurements of NO in CH(4)-air flames seeded with CH(3)Cl and CH(2)Cl(2) are described. The measurements are perturbed by strong photochemical effects characterized by UV emissions. The contribution of these background emissions is taken into account on the basis of an on-line-off-line resonance procedure. First results indicate an important increase of NO in the presence of chlorinated species. Background emissions observed in the range 220 260 nm and at 278 nm are ascribed, respectively, to electronically excited HCl and CCl photofragments. It is shown that C(2)H(3)Cl and CHCl(2) species are responsible for the formation of HCl and CCl, respectively, and a photolytic mechanism for formation of the photofragments is proposed. PMID- 18285966 TI - Measurements and modeling of acetone laser-induced fluorescence with implications for temperature-imaging diagnostics. AB - Recent determinations of the temperature dependence of acetone fluorescence have permitted the application of acetone planar laser-induced fluorescence imaging, which was already popular for mapping concentration, to the measurement of temperature. With a view toward developing temperature-imaging diagnostics, we present atmospheric-pressure fluorescence and absorption results acquired with excitation at eight wavelengths across the absorption feature of acetone and at temperatures from 300 to 1000 K. Modeling of the fluorescence yield of acetone is shown to be useful in explaining both these results and the variation of acetone fluorescence with pressure and composition that was observed in several studies. The model results in conjunction with the photophysics data provide guidance for the application of temperature diagnostics over a range of conditions while also suggesting useful multiparameter imaging approaches. PMID- 18285967 TI - Turn-key Raman lidar for profiling atmospheric water vapor, clouds, and aerosols. AB - We describe an operational, self-contained, fully autonomous Raman lidar system that has been developed for unattended, around-the-clock atmospheric profiling of water vapor, aerosols, and clouds. During a 1996 three-week intensive observational period, the system operated during all periods of good weather (339 out of 504 h), including one continuous five-day period. The system is based on a dual-field-of-view design that provides excellent daytime capability without sacrificing nighttime performance. It is fully computer automated and runs unattended following a simple, brief (~5-min) start-up period. We discuss the theory and design of the system and present detailed analyses of the derivation of water-vapor profiles from the lidar measurements. PMID- 18285968 TI - Water-core waveguide for pollution measurements in the deep ultraviolet. AB - A fiber optic system for water analysis with high transparency in the deep-UV region (lambda >/= 190 nm) is presented. The system consists of special UV improved silica fibers and a liquid-core waveguide (LCW) as an optical cell. The apertures of both light guides, the silica fiber and the LCW, are matched. The optical losses of the device are investigated experimentally and compared with theory, especially with a standard free-space geometry. The performance of the system with respect to UV absorption spectroscopy is demonstrated for nitrate and chlorine pollution in pure water. For a 203-mm-long LCW the detection limits have been determined to be as low as 22 mug/L for nitrate and 26 mug/L for chlorine. PMID- 18285969 TI - Applicability Limits of Beer's Law for Dispersion Media with a High Concentration of Particles. AB - This study analyzes the values of volume concentrations of scatterers at which radiation extinction in dispersion media obeys Beer's law. The dependence of the maximum particle concentration at which Beer's law holds on the properties of the dispersion medium is investigated. It is shown that the maximum concentration is strongly dependent on the scatterers' parameters and varies over a wide range, from tenths to tens of percent. PMID- 18285970 TI - Measurement of beam-shape coefficients in the generalized lorenz-mie theory for the on-axis case. AB - The use of the generalized Lorenz-Mie theory that describes the interaction between a spherical particle and an arbitrarily shaped beam requires knowledge of the beam-shape coefficients (BSC's) that describe the illuminating beam. Classically, these BSC's are evaluated from an a priori mathematical description of the illuminating beam. We propose a method that relies on intensity measurements along the beam axis that permits one to measure directly the BSC's of an actual beam in the laboratory. PMID- 18285971 TI - Optical free-path-length distribution in a fractal aggregate and its effect on enhanced backscattering. AB - A free-path-length distribution function (FPDF) of multiply backscattered light is theoretically derived for a fractal aggregate of particles. An effective mean free path-length l(D) is newly introduced as a measure of randomness analogous with a homogeneously random medium. We confirm the validity of the FPDF by demonstrating agreement between the dimensions designed for a particle distribution generated by a random walk based on the derived FPDF and estimated by the radius of gyration method. The FPDF is applied to Monte Carlo simulations for copolarized multiply backscattered light from the fractal aggregate of particles. It is shown that a copolarized intensity peak of enhanced backscattering in the far field decreases in accordance with theta(2-D) and has an angular width of lambda/l(D). This spatial feature of the backscattering enhancement corresponds to that of the copolarized intensity peak produced from a homogeneously random medium with a dimension of D = 3. As a result, the validity of the model for the fractal structure of particle aggregates and the applicability of the derived FPDF are confirmed by the numerical results. PMID- 18285972 TI - Anomalous diffraction theory for arbitrarily oriented finite circular cylinders and comparison with exact T-matrix results. AB - A general method is developed to formulate extinction and absorption efficiency for nonspherical particles at arbitrary and random orientations by use of anomalous diffraction theory (ADT). An ADT for finite circular cylinders is evaluated as an example. Existing ADT's for infinite cylinders at arbitrary orientations and for finite cylinders at the normal incidence are shown to be special cases of the new formulation. ADT solutions for finite cylinders are shown to approach the rigorous T-matrix results when the refractive indices approach unity. The importance of some physical processes that are neglected in the ADT approximation are evaluated by comparisons between ADT and rigorous calculations for different particle geometries. For spheres, van de Hulst's ADT and Mie theory are used, whereas the ADT that we present and T-matrix calculations are used for cylinders of different diameter-to-length ratios. The results show that the differences in extinction between ADT and exact solutions generally decrease with nonsphericity. A similar decrease occurs for absorption at wavelengths of relatively strong absorption. The influence of complex refractive index is evaluated. Our results suggest that ADT may provide a useful approximation in parameterization and remote sensing of cirrus clouds in the Christiansen bands where the real part of the refractive index approaches unity and/or where relative absorption is strong. PMID- 18285973 TI - Use of satellite natural vibrations to improve performance of free-space satellite laser communication. AB - In some of the future laser communication satellites, it is plausible to assume that tracking and communication receivers will use the same detector array. The reason for dual use of the detector is to design simpler and less expensive satellites. Satellites vibrate continually because of their subsystems and environmental sources. The vibrations cause nonuniform spreading of the received energy on the detector array. In view of this, the information from the tracking system is used to adapt individually the communication signal gain of each of the detectors in the array. This adaptation of the gains improves communication system performance. It is important to emphasize that the communication performance improvement is achieved only by gain adaptation. Any additional vibrations decrease the tracking and laser pointing system performances, which decrease the return communication performances (two-way communication). A comparison of practical communication systems is presented. The novelty of this research is the utilization of natural satellite vibrations to improve the communication system performance. PMID- 18285974 TI - Measure of the influence of detector noise on temperature-measurement accuracy for multiband infrared systems. AB - The noise-equivalent temperature difference is a measure of the detector-noise limited sensitivity of single-band IR systems for noncontact temperature measurement. However, because its definition is based on the signal-to-noise ratio in a single detector channel, the notion of noise-equivalent temperature difference must be generalized in case of dual-band or multiband IR systems. A new measure of temperature-measurement sensitivity is proposed that can be used to describe single-band, dual-band, and multiband IR measurement systems. With this measure a comparison of temperature-measurement accuracy among single-band, dual-band, and multiband systems was carried out. PMID- 18285975 TI - GaN-Based Solar-Ultraviolet Detection Instrument. AB - We report on the fabrication of a solar-UV monitoring system that uses GaN-based photodetectors. GaN photoconductors, p-n junction photodiodes, and Schottky barrier photodiodes have been fabricated and characterized as UV sensors. The best performances are obtained in Schottky photodiodes, which show a linear response, a flat responsivity of 100 mA/W, a visible rejection ratio higher than 10(3), and a noise-equivalent power of 1 nW/Hz(-1/2). Preliminary data on Al(x)Ga(1-x)N (x = 0.15, 0.22) detectors are also presented. Using GaN Schottky diodes, we fabricate and evaluate a complete solar-UV detection head. PMID- 18285976 TI - Variable-spacing diffraction grating employing elastomeric surface waves. AB - Variable-spacing diffraction gratings may be useful for controlling the deflection of a light beam. We introduce a method for making such a variable grating through the use of variable-frequency surface waves in elastomeric substrates. Elastomers have an extremely low Young's modulus compared with that of conventional solids, which allows surface waves to be created with low power and to travel much more slowly than waves in conventional surface-wave devices. To date, the best results have been obtained with a thin silicone elastomer membrane supported upon the surface of water. It has been found that the propagation speed of such waves is consistent with that in a previously developed analytical model, which predicts the dispersion relationship in terms of the membrane's thickness, tension, and modulus. Such a system has been used to cause a variable deflection of a light beam, in agreement with numerical predictions. PMID- 18285977 TI - Narcissistic Ghosts in Rowland-Mounted, Concave Gratings with nu = 0 degrees : a Cautionary Note. AB - Narcissistic ghost lines in a Rowland-grating spectrograph have been identified as arising from the reimaging of bright spectral features, which are incident on the face of a detector photocathode [Cesium iodide (CsI) on a microchannel plate], back onto the detector by the grating in zero order. The mean of the wavelength of the diffracted light and the apparent wavelength of the ghost allows the angle of the grating normal with respect to the input beam (alpha) to be determined. Measurements of ghost intensity as a function of wavelength are presented and are found to range between 7 x 10(-4) and 7 x 10(-3) of the parent line. We find that the sum of the CsI photocathode reflectivity and quantum efficiency <1/2, showing the bulk of the light incident upon the detector, is neither reflected nor detected. We caution that any Rowland circle spectrograph with a detector normal nearly aligned with the grating normal and with a sufficiently reflective detector face (or surrounding mounting structure) will suffer from these narcissistic ghosts. PMID- 18285978 TI - Crossed phase gratings with diffractive optical elements. AB - Orienting two identical or complementary diffractive gratings with a small angle between the grating grooves allows a new crossed-grating device to be constructed. This device has an effective profile that varies locally. For understanding the effects of this variation and the diffraction efficiency of the gratings, the local profiles were correlated with the moire period of the crossed grating system by use of various techniques. Asymmetric intensity behavior in the first order of the crossed gratings was seen. Effectively, the diffraction efficiency of the crossed gratings yielded a response equivalent to that of a grating with variable blaze that could be useful in optical computing as a passive optical switching device. One of several models is described that creates greater asymmetric behavior. PMID- 18285979 TI - Effects of Gap Width on Vacuum-Ultraviolet Transmission Through Submicrometer Period, FreeStanding Transmission Gratings. AB - The effects of gap width on the transmission coefficient of vacuum-ultraviolet light through submicrometer-period, freestanding transmission gratings are reported. Results from computations and an analytical waveguide model are shown to be consistent with experimental measurements. These results show that thin gratings with narrow gaps and thick gratings with wide gaps are equally effective at eliminating 121.6-nm radiation. The thin gratings with the narrow gaps have the advantage of better attenuation of shorter-wavelength radiation than the thick gratings with the larger gaps. PMID- 18285980 TI - Ion-exchanged diffractive elements in glass for substrate-mode optics. AB - We recently demonstrated the use of continuous-phase ion-exchanged diffractive elements in glass for free-space optics. We extend our design methods to substrate-mode optics, which permits compact packing of miniature-sized free space optical systems. We designed one-dimensional gratings for equal-intensity 1 ? 3 and 1 ? 5 beam splitting, assuming both planar and conical incidence angles. An experimental demonstration of a 1 ? 3 beam splitter with a uniformity error of 3.4% is presented. PMID- 18285981 TI - Coupling characteristics of cladding modes in tilted optical fiber bragg gratings. AB - We have studied both theoretically and experimentally the effect of grating tilting on the coupling between the fundamental core mode and the cladding modes in an optical fiber Bragg grating. The coupling is shown to be very sensitive on the tilting angle. It is also shown that tilting angle has to be minimized in fibers with designs to suppress the coupling between the fundamental core mode and the cladding modes. We have also studied the single, strong loss peak accompanying the Bragg reflection peak in depressed-cladding fibers, thus showing a good agreement between behavior that is measured and that is predicted theoretically. PMID- 18285982 TI - Analysis of Inhomogeneous Optical Systems by the Use of Ray Tracing. II. Three dimensional Systems with Symmetry. AB - We describe a new approach to the index reconstruction of three-dimensional optical systems with rotational symmetry, which is based on sampling ray paths that lie in the sagittal plane. Since the observed rays are distorted by the optical system itself, they cannot be used directly for index reconstruction. We present an iterative procedure to compute the true ray paths and then to find the index distribution. The utility of the method is verified on the model problem. PMID- 18285983 TI - Ellipsometric Scatterometry for the Metrology of Sub-0.1- num-Linewidth Structures. AB - We describe a modification to our existing scatterometry technique for extracting the relative phase and amplitude of the electric field diffracted from a grating. This modification represents a novel combination of aspects of ellipsometry and scatterometry to provide improved sensitivity to small variations in the linewidth of subwavelength gratings compared with conventional scatterometer measurements. We present preliminary theoretical and experimental results that illustrate the possibility of the ellipsometric scatterometry technique providing a metrology tool for characterizing sub-0.1-mum-linewidth. PMID- 18285984 TI - Determining the optical constants of read-write sliders during flying-height testing. AB - Flying-height testers for rigid disk drives employ a transparent glass substrate in place of the magnetic disk and use optical interferometry to measure the flight properties of the read-write slider. Because of the material phase change on reflection, the effective optical constants n and k of the slider play an important role in the measurement. We describe an instrument that determines the optical constants simultaneously with flying height, using polarization interferometry. This in situ analysis of n and k obviates the need for independent ellipsometry, while avoiding the problematic retract calibration characteristic of traditional flying-height test equipment. The rms uncertainty for n and k are 0.04, resulting in height uncertainties that range from 3 nm for 250-nm flying heights down to 0.5 nm at contact. We verify these results by use of a variety of experimental techniques on both laboratory samples and actual read-write sliders. PMID- 18285985 TI - Superluminescent diode interferometer using sinusoidal phase modulation for step profile measurement. AB - We propose an interferometer in which the relationship between the degree of coherence (DCH) and the optical path difference (OPD) is utilized for determining an OPD longer than a wavelength. A superluminescent diode is employed as the source of the interferometer, and sinusoidal phase-modulating interferometry is used to detect the DCH and the phase of the interference signal. The combination of the OPD determined from the DCH and the phase of an interference signal enables us to measure an OPD longer than a wavelength with a high accuracy of a few nanometers. Experimental results show clearly the usefulness of the interferometer for a step-profile measurement. PMID- 18285986 TI - Dispersion in stellar interferometry: simultaneous optimization for delay tracking and visibility measurements. AB - In long-baseline optical stellar interferometry, it is necessary to maintain optical path equality between the two arms of an interferometer in order to measure the fringe visibility. There will be errors in matching the optical paths because of a number of factors, and it is desirable to use an automatic system to monitor and correct such path errors. One type of system is a delay tracker, based on imaging of the channeled spectrum. The tracking algorithm is designed to maintain a fixed number of fringes, ideally linearly spaced, across the observed spectral band. This results in a constant optical path difference, which may be incompatible with the requirement of path equality for the measurement of fringe visibility. In a practical interferometer that uses an optical path-length compensator operating in air, there is a complication since air paths introduce differential dispersion. This dispersion can be compensated for by including dispersion correction. By modifying the operation of an appropriately designed dispersion corrector, we show that it is possible to make the optical path difference zero at the measurement wavelength and, at the same time, to produce linearly spaced channel fringes across the tracking band. PMID- 18285987 TI - Simultaneous measurement of diameter and position of spherical particles in a spray by an original imaging method. AB - The size and the position of individual particles in a falling spray crossing a horizontal plane are measured simultaneously with a shadowgraph imaging setup. An original method for measurement of drop sizes based on a prescribed analytical form of the luminance distribution of the objects (i.e., the drops) is presented. The processing time is strongly reduced compared with that for the usual inversion techniques. The turbulent dispersion of drops in a grid-generated turbulence is studied. Evidence for the presence of coalescence in the spray is presented. PMID- 18285988 TI - Spectroellipsometric method for process monitoring semiconductor thin films and interfaces. AB - Real-time monitoring by multiwavelength phase-modulated ellipsometry of the growth of plasma-deposited microcrystalline silicon (muc-Si) is presented. We discuss the construction of a growth model for process monitoring, and, in particular, we treat the inhomogeneity in the muc-Si layer by using an approximation of the reflection coefficient known as the WKBJ method. By also using the Bruggeman effective medium theory to describe the optical properties of muc-Si, we demonstrate monitoring the crystallinity in the upper and the lower part of the layer together with the thickness. The inversion algorithms thus remain very fast, with calculation times within 5 s on a standard Pentium computer. This makes possible precise control of the thickness and the crystallization of both the top and the bottom interface of the layer during the elaboration of devices such as solar cells and thin-film transistors. PMID- 18285989 TI - Polarization phase stepping with a savart element. AB - The imaging properties of a real-time shearing interferometer are presented. The use of Savart elements, both as a beam displacer and an analyzer in a polarization phase-stepping scheme, is demonstrated in a real-time, two-camera, four-bucket shearing interferometer. A simple calculation scheme for ray propagation through uniaxial, birefringent elements is presented, and the effects on the image formation through 6-cm-long Savart elements is discussed. PMID- 18285990 TI - CATRAM: An Apparatus for the Optical Characterization of Advanced Transparent Materials. AB - At the Transparent Materials Laboratory of the Italian Agency for Energy, New Technologies and Environment, the characterization of advanced transparent materials (CATRAM) facility permits the optical characteristics of transparent advanced materials to be evaluated, a procedure for which spectrophotometers are not adequate. The CATRAM facility is based mainly on a large-diameter integrating sphere to perform transmittance, reflectance, and absorptance measurements. We describe the measurement procedures and verify the reliability of the facility by comparing the results obtained for several samples with those from other laboratories. Furthermore, some system implementations for improvement of accuracy are discussed. PMID- 18285991 TI - Dual-hologram shearing interferometry with regulated sensitivity. AB - A novel optical diagnostic technique, namely, dual-hologram shearing interferometry with regulated sensitivity, is proposed for visualization and measuring of the density gradients of compressible flows in wind tunnels. It is superior to conventional shearing interferometry in both accuracy and sensitivity. The method is especially useful for strong turbulent or unsteady regions of the flows, including shock flows. The interferometer has proved to be insensitive to mechanical vibrations and has allowed us to record holograms during the noisy wind-tunnel run. The proposed approach is demonstrated by application to a supersonic flow over spherically blunted and sharp nose-cone cylinder models. We believe that the technique will become an effective tool for receiving optical data in many flow facilities. PMID- 18285992 TI - Measurement of the complex refractive-index spectrum for birefringent and absorptive liquids. AB - The optical constants of birefringent and/or opaque liquids, e.g., liquid crystals and magnetic fluids, are difficult to measure at wavelengths at which a strong light source such as a laser or an arc lamp is not accessible. The refractive index n and the extinction coefficient kappa of these liquids can be simultaneously evaluated from the reflectance curves that are measured in the large incident angle range. A semicylindrical sample cell allows the spectral reflectance measurement with a weak light source even at large incident angles. By using this method, we evaluated the ordinary and the extraordinary indices of a nematic liquid crystal in the continuous wavelength range of 0.55-1.60 mum. The complex refractive indices of magnetic fluids were also evaluated, and the affect of the magnetic field was demonstrated. PMID- 18285993 TI - Holographic interferometers with photorefractive recording media. AB - We discuss two types of holographic interferometer that contain photorefractive recording media. The first type contains two beams interacting in a photorefractive medium. The second type utilizes a single beam and relies on self pumped phase conjugation from a photorefractive crystal to make phase changes appear as intensity changes. We show both theoretically and experimentally that the first type can be analyzed in a straightforward manner; however, the second type cannot be approximated as simply a special case of the first type, as one may naively suspect. PMID- 18285994 TI - Phase-aberration correction with dual liquid-crystal spatial light modulators. AB - A phase-aberration-correction system that uses high-resolution, twisted nematic liquid-crystal spatial light modulators in a Mach-Zehnder interferometer is presented. A correction algorithm is described and experimentally verified by use initially of one liquid-crystal panel. Phase aberrations are successfully removed by a single liquid-crystal panel, but unacceptably high amplitude variation is introduced into the wave front because of the phase-amplitude coupling of the spatial light modulator. A second panel is used to remove the amplitude modulation. The modified optical system with a multiplicative architecture is described, and results are presented that show the correction of phase aberrations with an amplitude variation of less than 10%. PMID- 18285995 TI - Stability of systems of plane reflecting surfaces. AB - A general rule for the stability of plane reflecting surface systems is derived by use of the features of the reflection matrix. It is proved that only two directions can be stable: the forward direction and the backward direction (retroreflection). Examples for the application of this rule in the design of stable reflecting systems for optical communication are given. PMID- 18285996 TI - Exact Design of Aplanatic Microscope Objectives consisting of Two Conic Mirrors. AB - The necessary equations are derived for the design of aplanatic microscope objectives consisting of two mirrors, one concave and the other convex. The first order parameters are calculated along with the conic constants of the mirrors, determined by means of an exact analysis to arrive at an aplanatic system. PMID- 18285997 TI - Computer numerically controlled plasma chemical vaporization machining with a pipe electrode for optical fabrication. AB - We have developed a chemical vaporization machining device that has computer numerically controlled plasma, by using a pipe electrode for optical fabrications. In this device, less than approximately 1 atm of pressure, plasma is generated around the tip of a pipe electrode. During the process, a workpiece is scanned against the electrode under computer control to achieve the desired shape to be removed. A workpiece of silica glass plate is shaped by use of this device, and the removal characteristics of the device are examined. The equations to characterize numerically the shape resulting from scanning of a workpiece have been derived. The new device allows the high precision of optics from the micrometer to the nanometer level with high-speed removal. The shaped surface is sufficiently smooth to be suitable for optical use. PMID- 18285998 TI - Making fast cylindrical gradient-index lenses diffraction limited by using a wave front-correction element. AB - A wave-front-correction element (WFCE) is produced to make a cylindrical Ag-ion exchanged gradient-index (GRIN) lens with a high numerical aperture (0.53) diffraction limited (wave-front error, 0.02lambda rms). The wave-front aberrations of the cylindrical GRIN lens are measured by a phase-shifting shearing interferometer, with a conventional microscope objective used as a compensation lens. The continuous surface relief of the WFCE is produced by a lithographic process. The wave-front-corrected GRIN lens is applied to collimate the strongly divergent light (57 degrees full diverging angle measured at 1/e(2) of maximum intensity) emitted by a high-power diode laser. The power irradiated into a full angle of 2 mrad can be enhanced by a factor of 1.8 with the WFCE. PMID- 18285999 TI - Design and optimization of an irradiance profile-shaping system with a genetic algorithm method. AB - We develop a genetic algorithm (GA) optimization method and use it in the design of a refractive-beam profile-shaping system. In this application, we employ the GA to determine the shape of one surface of the primary beam profile-shaping element in our system. The GA is instructed to vary the shape of this surface such that the output intensity profile is flat on a spherical surface some distance away. The GA does this while insuring that only a specified area of the output surface is illuminated. The calculation of the intensity profile is based on geometrical optics and is accomplished exclusively through ray tracing, giving this method broad applicability. PMID- 18286000 TI - Optical acceptance function of modified compound parabolic concentrators with linear corrugated reflectors. AB - For reflective cylindrical concentrators the acceptance function, as determined in the meridian plane, remains constant for more oblique planes of incidence. However, if the reflective surface has a linear corrugated structure, it is possible to increase the acceptance in the meridian plane at the expense of reduced acceptance at a more oblique incidence. A ray-tracing study has been performed on the optical properties of troughlike compound parabolic concentrators (CPC's) with linear corrugated reflectors. Calculations of angular acceptance for a modified CPC with concentration C = x is presented. A possible application of this new concentrator is as a secondary concentrator for tracking parabolic troughs. PMID- 18286001 TI - High-efficiency prism light guides with confocal parabolic cross sections. AB - Prism light guides are often used for transporting and distributing light. Typically such guides are tubular structures lined with a flexible prismatic sheet material that confines light by means of total internal reflection. Inasmuch as the prismatic material is produced as a flat sheet, such guides require at least one seam in formation of the tubular structure, and the optical imperfection associated with seams is often a dominant loss mechanism. We present a new configuration for the cross-sectional shape of a prism light guide that substantially reduces the loss caused by such seams. PMID- 18286002 TI - Polarization-based tunable interferometric filter. AB - A polarization-based tunable interferometric filter essentially consisting of a two-beam interferometer with birefringence elements is described. The analysis of the filter is done through the concept of a geometric phase in optics-namely, the Pancharatnam phase. The transmission characteristics of the filter can be controlled through three parameters: the thickness of the birefringent elements, the optical path difference, and the orientation angle of an analyzer placed at the interferometer output. It is demonstrated theoretically that, with a particular choice of these parameters, the chromatic dispersion of the filter is compensated in a given spectral range. Some properties of the device are confirmed by an experimental demonstration. PMID- 18286003 TI - Effect of surface roughness and subsurface damage on grazing-incidence x-ray scattering and specular reflectance. AB - Grazing-incidence specular reflectance and near-specular scattering were measured at Al-K(alpha) (1.486-keV, 8.34-?) radiation on uncoated dielectric substrates whose surface topography had been measured with a scanning probe microscope and a mechanical profiler. Grazing-incidence specular reflectance was also measured on selected substrates at the Cu-K(alpha) (8.047-keV, 1.54-?) wavelength. Substrates included superpolished and conventionally polished fused silica; SiO(2) wafers; superpolished and precision-ground Zerodur; conventionally polished, float polished, and precision-ground BK-7 glass; and superpolished and precision-ground silicon carbide. Roughnesses derived from x-ray specular reflectance and scattering measurements were in good agreement with topographic roughness values measured with a scanning probe microscope (atomic force microscope) and a mechanical profiler that included similar ranges of surface spatial wavelengths. The specular reflectance was also found to be sensitive to the density of polished surface layers and subsurface damage down to the penetration depth of the x rays. Density gradients and subsurface damage were found in the superpolished fused-silica and precision-ground Zerodur samples. These results suggest that one can nondestructively evaluate subsurface damage in transparent materials using grazing-incidence x-ray specular reflectance in the 1.5-8-keV range. PMID- 18286004 TI - High-resolution monochromatic x-ray imaging system based on spherically bent crystals. AB - We have developed an improved x-ray imaging system based on spherically curved crystals. It is designed and used for diagnostics of targets ablatively accelerated by the Nike KrF laser. A spherically curved quartz crystal (d = .?, R = mm) has been used to produce monochromatic backlit images with the He-like Si resonance line (1865 eV) as the source of radiation. The spatial resolution of the x-ray optical system is 1.7 mum in selected places and 2-3 mum over a larger area. Time-resolved backlit monochromatic images of polystyrene planar targets driven by the Nike facility have been obtained with a spatial resolution of 2.5 mum in selected places and 5 mum over the focal spot of the Nike laser. PMID- 18286005 TI - Optical characterization of dielectric and semiconductor thin films by use of transmission data. AB - A method to calculate the optical functions n(lambda) and k(lambda) by use of the transmission spectrum of a dielectric or semiconducting thin film measured at normal incidence is described. The spectrum should include the low-absorption region and the absorption edge to yield the relevant optical characteristics of the material. The formulas are derived from electromagnetic theory with no simplifying assumptions. Transparent films are considered as a particular case for which a simple method of calculation is proposed. In the general case of absorbing films the method takes advantage of some properties of the transmittance T(lambda) to permit the parameters in the two regions mentioned above to be calculated separately. The interference fringes and the optical path at the extrema of T(lambda) are exploited for determining with precision the refractive index and the film thickness. The absorption coefficient is computed at the absorption edge by an efficient iterative method. At the transition zone between the interference region and the absorption edge artifacts in the absorption curve are avoided. A small amount of absorption of the substrate is allowed for in the theory by means of a factor determined from an independent measurement, thus improving the quality of the results. Application of the method to a transmission spectrum of an a:Si(x)N(1-x):H film is illustrated in detail. Refractive index, dispersion parameters, film thickness, absorption coefficient, and optical gap are given with the help of tables and graphs. PMID- 18286006 TI - Optical properties of metallic films for vertical-cavity optoelectronic devices. AB - We present models for the optical functions of 11 metals used as mirrors and contacts in optoelectronic and optical devices: noble metals (Ag, Au, Cu), aluminum, beryllium, and transition metals (Cr, Ni, Pd, Pt, Ti, W). We used two simple phenomenological models, the Lorentz-Drude (LD) and the Brendel-Bormann (BB), to interpret both the free-electron and the interband parts of the dielectric response of metals in a wide spectral range from 0.1 to 6 eV. Our results show that the BB model was needed to describe appropriately the interband absorption in noble metals, while for Al, Be, and the transition metals both models exhibit good agreement with the experimental data. A comparison with measurements on surface normal structures confirmed that the reflectance and the phase change on reflection from semiconductor-metal interfaces (including the case of metallic multilayers) can be accurately described by use of the proposed models for the optical functions of metallic films and the matrix method for multilayer calculations. PMID- 18286007 TI - Repeatability of the composition of titanium oxide films produced by evaporation of ti 2 o 3. AB - Congruent vaporization is a process that yields constant vapor species. Ti(2)O(3) was continuously electron-beam evaporated to produce titanium oxide thin films. Rutherford backscattering spectrometry was employed to study the evolution of the composition of these films. It seems that congruent vaporization can be established in a coating plant. TiO(2) films produced by conventional reactive deposition tend to contain mixtures of titanium oxides. Increasing the transmission of TiO(2) films becomes an issue of increasing the TiO(2) component in the films by adequate reactive evaporation. PMID- 18286008 TI - Modeling the index of refraction of insulating solids with a modified lorentz oscillator model. AB - A modification of the Lorentz oscillator model for optical constants is proposed in an effort to achieve better agreement with experimental data while keeping the calculation simple. Improvement in agreement between theoretical and experimental data obtained with a variable line shape (frequency-dependent damping constant) over a wide spectral range is demonstrated through modeling the index of refraction of Si(3)N(4) (1-24 eV), SiO (0.15-25 eV) and amorphous and crystalline SiO(2) (0.15-25 eV). Model parameters are estimated by acceptance-probability controlled simulated annealing. Excellent agreement between the modified model and the experimental data is obtained for both real and imaginary parts of the index of refraction. PMID- 18286009 TI - Fast characterization of metal films for fiber attenuators. AB - We propose a new procedure for evaluating the complex refractive index of a metal film, based on transmission measurements at different incidence angles. The method is simpler and faster than standard ellipsometry and performs the accuracy required for the design of fiber-optic attenuators for telecommunications. As an example, we report on a device showing a constant attenuation on the 1200-1600-nm wavelength range. PMID- 18286010 TI - Efficient High-Energy Pulse-Train Generation Using a 2 n-Pulse Michelson Interferometer. AB - We demonstrate a novel, Michelson-based, ultrafast multiplexer with a throughput approaching 100% for a polarization-multiplexed train and 50% for a linearly polarized train, which is compatible with a high-energy pulse train and shaped pulse generation. The interpulse spacings in the resultant 2(n)-pulse train can be adjusted continuously from multinanoseconds through zero. Using this interferometer, we also demonstrate generation of a 16-pulse train of terahertz pulses. PMID- 18286011 TI - Coherence-based imaging through turbid media by use of degenerate four-wave mixing in thin liquid-crystal films and photorefractives. AB - We describe a coherence-based imaging technique based on degenerate four-wave mixing in two materials. First, a 45 degrees -cut BaTiO(3) crystal was used as a self-pumped phase conjugator to obtain depth-resolved images through a 4 mean free-path (mfp) scattering media. In addition, an HITC dye-doped K15 liquid crystal layer was used to provide single-shot image acquisition through a 2 mfp scattering media. This technique can be used to provide instantaneous (single pulse), depth-resolved, two-dimensional images of the internal structure of scattering materials. Potential applications of this technique include subsurface imaging of biomedical tissue and nondestructive evaluation of composite materials. PMID- 18286012 TI - Optical diffusion of continuous-wave, pulsed, and density waves in scattering media and comparisons with radiative transfer. AB - We discuss several outstanding theoretical problems in optical diffusion in random media. Specifically, we discuss which of several diffusion theories most closely approximates exact solutions of the equation of transfer. We consider a plane wave impinging upon a plane-parallel slab of a random medium as a model problem to compare the diffusion theories with a numerical solution of the equation of transfer for continuous-wave, pulsed, and photon density waves. In addition, we discuss the validity of the diffusion approximation for a variety of parameter settings to ascertain the diffusion approximation's applicability to imaging biological media. PMID- 18286013 TI - Microscopic imaging through a turbid medium by use of annular objectives for angle gating. AB - We report a new method for microscopic imaging of an object embedded in a turbid medium. The new method is based on the angle-gating mechanism achieved by the use of polarized annular objectives in the illumination and collection paths of a microscopic imaging system. A detailed experimental study is presented of the effects of the size of annular obstructions on image quality when turbid media, including polystyrene microspheres and milk suspensions, are imaged. Images of 22 mum polystyrene microspheres embedded in the turbid media show that misinterpretation can occur when circular objectives are used, because of the detection of mainly multiply scattered photons (i.e., diffusing photons). However, when annular objectives are employed, diffusing photons from a turbid medium can be efficiently suppressed; thus image contrast appears correctly, and image resolution is increased. PMID- 18286014 TI - Two-dimensional near-infrared transillumination imaging of biomedical media with a chromium-doped forsterite laser. AB - Transillumination images of objects hidden in normal and cancerous human breast tissues and bovine, porcine, and gallinaceous (chicken) tissues as well as model random-scattering media were recorded with 1250-nm light from a chromium-doped forsterite laser. A Fourier space gate and a polarization gate were used to sort out image-bearing photons and discriminate against multiply scattered image blurring photons. Better contrast, higher spatial resolution, and deeper penetration of samples were achieved for imaging with 1250-nm light than those obtained at shorter wavelengths, such as 1064 nm from a neodymium-doped yttrium aluminum garnet (YAG) laser. Better contrast and higher resolution were also obtained when the object was imaged through normal human breast tissue than through cancerous breast tissue. Images with marked distinction between fatty and fibrous human breast tissues were obtained when the Cr:forsterite laser was tuned to 1225 nm, a wavelength that resonates with an optical absorption band of breast fat tissues. Imaging with linearly polarized light revealed that the image quality depends significantly on the orientation of the polarization of the incident light with respect to the fibers in the bovine tissue. PMID- 18286015 TI - Frequency-domain fluorescent diffusion tomography: a finite-element-based algorithm and simulations. AB - We present a finite-element-based algorithm for reconstruction of fluorescence lifetime and yield in turbid media, using frequency-domain data. The algorithm is based on a set of coupled diffusion equations that describe the propagation of both excitation and fluorescent emission light in multiply scattering media. Centered on Newton's iterative method, we implemented our algorithm by using a synthesized scheme of Marquardt and Tikhonov regularizations. A low-pass spatial filter is also incorporated into the algorithm for enhancing image reconstruction. Simulation studies using both noise-free and noisy data have been performed with the nonzero photon density boundary conditions. Our results suggest that quantitative images can be produced in terms of fluorescent lifetime and yield values and location, size, and shape of heterogeneities within a circular background region. PMID- 18286016 TI - Absolute Emission Spectra from Bacillus subtilis and Escherichia coli Vegetative Cells in Solution. AB - Spectrally resolved emission (270-560 nm) from dilute suspensions of washed Bacillus subtilis and Escherichia coli were measured by use of tunable laser excitation between 270 and 300 nm. Integrated absolute emission cross sections increase with decreasing excitation wavelength and range from 1.8 x 10(-12) to 6.0 x 10(-11) cm(2)/(particle sr). An emission band near 340 nm dominates all observed spectra. At each excitation wavelength spectrally resolved emissions from the E. coli and B. subtilis suspensions are indistinguishable. PMID- 18286017 TI - Control of chromatic focal shift through wave-front coding. AB - Control of chromatic aberration through purely optical means is well known. We present a novel, to our knowledge, optical-digital method of controlling chromatic aberration. The optical-digital system, which incorporates a cubic phase-modulation (CPM) plate in the optical system and postprocessing of the detected image, effectively reduces a system's sensitivity to misfocus in general or axial (longitudinal) chromatic aberration, in particular. A fully achromatic imaging system (one that is corrected for a continuous range of wavelengths) can be achieved by initial optimization of the optical system for all aberrations except chromatic aberration. The chromatic aberration is corrected by the inclusion of the CPM plate and postprocessing. PMID- 18286018 TI - Design and operation of an in situ microchannel alignment-detection system. AB - The design of an alignment-detection system that uses off-axis diffractive elements and photodetectors is presented. The system was developed to detect the real-time misalignment of an array of optical beams as they pass through a microchannel relay. The design of this scheme is presented along with experimental results obtained from a prototype detection system. PMID- 18286019 TI - Balancing interpixel cross talk and detector noise to optimize areal density in holographic storage systems. AB - We investigate the effects of interpixel cross talk and detector noise on the areal storage density of holographic data storage. A numerical simulation is used to obtain the bit-error rate (BER) as a function of hologram aperture, pixel fill factors, and additive Gaussian intensity noise. We consider the effect of interpixel cross talk at an output pixel from all possible configurations of its 12 closest-neighbor pixels. Experimental verification of this simulation procedure is shown for several fill-factor combinations. The simulation results show that areal density is maximized when the aperture coincides with the zero order of the spatial light modulator (SLM) (Nyquist sampling condition) and the CCD fill factor is large. Additional numerical analysis including finite SLM contrast and fixed-pattern noise show that, if the fixed-pattern noise reaches 6% of the mean signal level, the SLM contrast has to be larger than 6:1 to maintain high areal density. We also investigate the improvement of areal density when error-prone pixel combinations are forbidden by using coding schemes. A trade-off between an increase in areal density and the redundancy of a coding scheme that avoids isolated-on pixels occurs at a code rate of approximately 83%. PMID- 18286020 TI - Phasor-Difference Method for Measuring the Amplitude and the Phase of Small Amplitude Vibrations with TV Holography. AB - A new, to the author's knowledge, method for time-averaged TV holography measurements of small-amplitude vibrations is presented. In time-averaged TV holography with sinusoidal phase modulation of the reference arm of the interferometer, two phasors describe the object vibration and the modulation of the reference arm. By inversion of the squared zero-order Bessel function of the first kind, it is possible to measure the distance between these two phasors. The distances from an object-vibration phasor to a number of known reference phasors are measured to determine the amplitude and the phase of the object vibration. The method is demonstrated by the measurement of a vibration mode of a circular metal disk. The results are compared with theoretical data and with data obtained by a commonly used method in phase-modulated TV holography. PMID- 18286021 TI - Pattern generation with an extended focal depth. AB - The depth of focus of light patterns can be extended, within given tolerances, beyond the classical limits. For a quantitative evaluation we introduce a degree of depth-of-focus extension and a three-dimensional energy-distribution efficiency. The basic limitations involved in depth-of-focus extension are discussed. A coherent system in which the input is optimized for a desired output pattern is presented. An example of a pattern containing diffraction-limited line segments and a 4 times improvement in depth of focus is demonstrated. This task is much more difficult than generating patterns of isolated light spots in which the depth of focus is extended beyond an order of magnitude. PMID- 18286022 TI - Planar-integrated talbot array illuminators. AB - We demonstrate the planar integration of Talbot array illuminators designed to generate one-dimensional spot arrays. The array illuminator basically consists of a phase grating and a cylindrical diffractive lens integrated as a single diffractive optical element onto a transparent glass substrate. We discuss various design aspects, and we focus on problems typical for planar-integrated free-space optics like the tilted optical axis of the system. Experimental results and measurements, which were obtained from planar-integrated setups fabricated as surface-relief structures on a transparent glass substrate by use of standard photolithography, are included. PMID- 18286023 TI - Parallel joint transform correlator applied to devanagari script recognition. AB - A novel, to our knowledge, scheme for a parallel joint transform correlator system is demonstrated by the introduction of a five-channel zone-plate image multiplicator combined with photographic referencing and is applied to the recognition of Devanagari script. For considering the script's specific features, a partial image was obtained by means of windowing with 25 x 25 pixels out of a 64 x 64 pixel image of full-size characters and was employed for correlation. A comprehensive measurement of all 38 fundamental characters, namely, 1444 correlation pairs, showed satisfactory discrimination performance. PMID- 18286024 TI - Fast phase-unwrapping algorithm based on a gray-scale mask and flood fill. AB - Phase-unwrapping algorithms, an active and interesting subject in recent years, are important in a great number of measurement applications. Active research is being undertaken to develop reliable and high-speed procedures. The current process uses a gray-scale mask and the flood-fill concept from image processing for phase unwrapping. The algorithm unwraps phase from an area with higher reliability to one with lower reliability. In addition to robustness, the speed of the algorithm proposed is much faster than conventional routines. The experimental results of different algorithms are compared by analysis of a tooth plaster and a photoelastic specimen. PMID- 18286025 TI - Algebraic reconstruction of a small-scale wave front. AB - We show that a small-scale wave front can be reconstructed by an algebraic procedure from its intensity distribution in the focal plane, except for the ambiguities of the piston phase and the point-symmetrical solution of the complex conjugate. Details of the reconstruction procedure for a 3 x 3 wave front are presented, and the effectiveness of this procedure for a contaminated case is shown by computer simulation. A method for overcoming the ambiguity problem resulting from the point-symmetrical solution is also suggested. PMID- 18286026 TI - Parametric Wave-Aberration Retrieval from Point-Spread Function Data by use of a Pyramidal Recursive Algorithm. AB - A new, to my knowledge, procedure for retrieving the wave aberration from the point-spread function is presented. It uses the Levenberg-Marquardt optimization algorithm in a mutiresolution pyramidal scheme. The method, tested with simulated large aberrations without initial estimates, accelerates convergence and avoids stagnation in local minima. PMID- 18286027 TI - Experimental evaluation of user capacity in holographic data-storage systems. AB - An experimental procedure for determining the relation between the number of stored holograms and the raw bit-error rate (BER) (the BER before error correction) of a holographic storage system is described. Compared with conventional recording schedules that equalize the diffraction efficiency, scheduling of recording exposures to achieve a uniform raw BER is shown to improve capacity. The experimentally obtained capacity versus the raw-BER scaling is used to study the effects of modulation and error-correction coding in holographic storage. The use of coding is shown to increase the number of holograms that can be stored; however, the redundancy associated with coding incurs a capacity cost per hologram. This trade-off is quantified, and an optimal working point for the overall system is identified. This procedure makes it possible to compare, under realistic conditions, system choices whose impact cannot be fully analyzed or simulated. Using LiNbO(3) in the 90 degrees geometry, we implement this capacity-estimation procedure and compare several block-based modulation codes and thresholding techniques on the basis of total user capacity. PMID- 18286028 TI - Nonseparable two-dimensional fractional fourier transform. AB - Previous generalizations of the fractional Fourier transform to two dimensions assumed separable kernels. We present a nonseparable definition for the two dimensional fractional Fourier transform that includes the separable definition as a special case. Its digital and optical implementations are presented. The usefulness of the nonseparable transform is justified with an image-restoration example. PMID- 18286029 TI - Efficient detour-phase encoding of one-dimensional multilevel phase diffractive elements. AB - We show that detour-phase encoding with a multilevel blaze structure as a carrier grating is especially suited to the implementation of diffractive elements with relatively high complexity in one axis. For our proposal the carrier grating is aligned perpendicularly to this axis. In this way the element can be encoded with a high space-bandwidth product and a high phase resolution by use of a moderate carrier frequency. Moreover, this frequency can be adjusted to isolate the reconstructed field from the noise resulting from high diffraction orders of the carrier grating or caused by etching errors during fabrication. PMID- 18286030 TI - Design and characterization of a liquid-crystal spatial light modulator for a polarization-insensitive optical space switch. AB - We report preliminary results concerning a free-space optical switch between single-mode fibers with a ferroelectric liquid-crystal (FLC) spatial light modulator (SLM). In particular, we show experimentally that such a device can operate in a polarization-insensitive manner. The influence of the geometrical and physical features of the FLC SLM on the overall performance of the optical fiber switch are also discussed. PMID- 18286031 TI - Recirculating fiber delay-line filter with a fiber bragg grating. AB - We propose a new recirculating fiber delay-line filter structure. In the proposed system a fiber Bragg grating (FBG) is used as a multireflection mirror in a fiber delay loop, and an erbium-doped fiber is used to provide gain in the loop. By adjusting parameters such as the coupling efficiency, the reflectivity of the FBG, and the gain, we can change the characteristics of the filter transfer function. PMID- 18286032 TI - Phase-change visualization in two-dimensional phase objects with a semiderivative real filter. AB - A method of visualization of phase changes in two-dimensional pure-phase objects by use of two orthogonal Fourier plane filters that realize the half-order differentiation is presented. Real semiderivative filters used in two dimensions and in sequence yield output-image intensity signals proportional to the first derivatives of the input-object phase that appear on a constant background. This nonlinear filtration of spatial frequencies permits the alleviation of the consequences of square-law detection and makes phase changes visible. Phase changes in gradient-index phosphate glass are calculated experimentally. We discuss the accuracy of the proposed method. PMID- 18286033 TI - Compact polarization-based all-optical interconnection systems with growth capability. AB - All-optical communication requires all-optical interconnections, thus leading to reliable, fast, and flexible modular communication means in future systems. Free space approaches are advantageous since they fully use the two dimensions optics offer. A folded architecture based on a polarization code is proposed for dynamic optical interconnection. The suggested systems are compact and appropriate for both intracomputer and intercomputer communication. The modularity of the proposed architecture is presented, and a growth rule for the fully connected versions of the system is introduced. The proposed approach significantly reduces both the price of the interconnection systems and their complexity. Presented are 4 x 4 and 8 x 8 fully connected switches, a rearrangeable nonblocking 4 x 4 switch, and a crossbar architecture. PMID- 18286034 TI - Dynamic null steering in an ultrawideband time-steered array antenna. AB - We develop a method for forming squint-free wideband nulls in the antenna pattern of an ultrawideband array antenna. The technique uses an optical dispersive-prism beam former to provide time-delayed microwave signals to each antenna element for forming a squint-free main beam. The amplitude-modulated optical carrier is propagated through a set of optical links. Each link feeds an array element and includes an amount of dispersion proportional to element position. Tuning the wavelength of the optical carrier controls the microwave signal's arrival-time delay gradient across the array. A dispersive-prism tapped delay-line microwave filter is used to frequency shape a nulling signal. The wideband nulls do not significantly distort the main beam and are steered independently of the main beam. The technique is applied to sidelobe nulling for a transmitter and for jammer suppression for a receiver array. PMID- 18286035 TI - Polarization lidar returns from aerosols and thin clouds: a framework for the analysis. AB - Relationships for the interpretation of polarization lidar observations of aerosols and thin clouds are presented. They allow for the separation of contributions to backscatter from solid and liquid phases by the use of either the classical backscatter and depolarization ratio parameters or the particulate cross-polarized backscatter cross sections. It is shown that different aerosol phases can be better separated by use of the latter coordinates. Emphasis is placed on the study of composition and phase properties of polar stratospheric aerosols. PMID- 18286036 TI - Compact airborne lidar for tropospheric ozone: description and field measurements. AB - An airborne lidar has been developed for tropospheric ozone monitoring. The transmitter module is based on a solid-state Nd:YAG laser and stimulated Raman scattering in deuterium to generate three wavelengths (266, 289, and 316 nm) that are used for differential ozone measurements. Both analog and photon-counting detection methods are used to produce a measurement range up to 8 km. The system has been flown on the French Fokker 27 aircraft to perform both lower tropospheric (0.5-4-km) and upper tropospheric (4-12-km) measurements, with a 1 min temporal resolution corresponding to a 5-km spatial resolution. The vertical resolution of the ozone profile can vary from 300 to 1000 m to accommodate either a large-altitude range or optimum ozone accuracy. Comparisons with in situ ozone measurements performed by an aircraft UV photometer or ozone sondes and with ozone vertical profiles obtained by a ground-based lidar are presented. The accuracy of the tropospheric ozone measurements is generally better than 10-15%, except when aerosol interferences cannot be corrected. Examples of ozone profiles for different atmospheric conditions demonstrate the utility of the airborne lidar in the study of dynamic or photochemical mesoscale processes that control tropospheric ozone. PMID- 18286037 TI - Errors in the retrieval of thin-cloud optical parameters obtained with a two boundary algorithm. AB - The effect of various errors on the retrieval of optical depth, integrated backscatter, and extinction-to-backscatter ratio in optically thin clouds by use of a two-boundary algorithm is discussed. Uncertainties regarding aerosol loading at the cloud base and top lead to relevant errors that are often larger than those produced by signal noise. Formulas expressing the errors in the lidar derived optical quantities dependent on optical depth and aerosol uncertainties at the base and top are derived for different fitting procedures. A method for the reduction of errors in the case of consecutive cloud measurements is explored, consisting of the fitting of the retrieved optical-depth-integrated backscatter data to obtain a correct extinction-to-backscatter ratio. PMID- 18286038 TI - Satellite-sensor calibration verification with the cloud-shadow method. AB - An atmospheric-correction method appropriate for high-spatial-resolution sensors that uses cloud-shaded pixels together with pixels in a neighboring region of similar optical properties is described. This cloud-shadow method uses the difference between the total radiance values observed at the sensor for these two regions, thus removing the nearly identical atmospheric radiance contributions to the two signals (e.g., path radiance and Fresnel-reflected skylight). What remains is largely due to solar photons backscattered from beneath the sea to dominate the residual signal. Normalization by the direct solar irradiance reaching the sea surface and correction for some second-order effects provides the remote-sensing reflectance of the ocean at the location of the neighbor region, providing a known ground target spectrum for use in testing the calibration of the sensor. A similar approach may be useful for land targets if horizontal homogeneity of scene reflectance exists about the shadow. Monte Carlo calculations have been used to correct for adjacency effects and to estimate the differences in the skylight reaching the shadowed and neighbor pixels. PMID- 18286039 TI - Lidar in-space technology experiment measurements of sea surface directional reflectance and the link to surface wind speed. AB - The dependence of sea surface directional reflectance on surface wind stress suggests a method for deriving surface wind speed from space-based lidar measurements of sea surface backscatter. In particular, lidar measurements in the nadir angle range from 10 degrees to 30 degrees appear to be most sensitive to surface wind-speed variability in the regime below 10 m/s. The Lidar In-space Technology Experiment (LITE) shuttle lidar mission of September 1994 provided a unique opportunity to measure directional backscatter at selected locations by use of the landmark track maneuver and to measure fixed-angle backscatter from the ocean surfaces on a global scale. During the landmark track maneuver the shuttle orbiter orientation and roll axis are adjusted continuously to maintain the lidar footprint at a fixed location for a duration of ~1 min. Several data sets were converted to calibrated reflectance units and compared with a surface reflectance model to deduce surface wind speeds. Comparisons were made with ERS-1 scatterometer data and surface measurements. PMID- 18286040 TI - Atmospheric correction of ocean color imagery: use of the junge power-law aerosol size distribution with variable refractive index to handle aerosol absorption. AB - When strongly absorbing aerosols are present in the atmosphere, the usual two step procedure of processing ocean color data-(1) atmospheric correction to provide the water-leaving reflectance (rho(w)), followed by (2) relating rho(w) to the water constituents-fails and simultaneous estimation of the ocean and aerosol optical properties is necessary. We explore the efficacy of using a simple model of the aerosol-a Junge power-law size distribution consisting of homogeneous spheres with arbitrary refractive index-in a nonlinear optimization procedure for estimating the relevant oceanic and atmospheric parameters for case 1 waters. Using simulated test data generated from more realistic aerosol size distributions (sums of log-normally distributed components with different compositions), we show that the ocean's pigment concentration (C) can be retrieved with good accuracy in the presence of weakly or strongly absorbing aerosols. However, because of significant differences in the scattering phase functions for the test and power-law distributions, large error is possible in the estimate of the aerosol optical thickness. The positive result for C suggests that the detailed shape of the aerosol-scattering phase function is not relevant to the atmospheric correction of ocean color sensors. The relevant parameters are the aerosol single-scattering albedo and the spectral variation of the aerosol optical depth. We argue that the assumption of aerosol sphericity should not restrict the validity of the algorithm and suggest an avenue for including colored aerosols, e.g., wind-blown dust, in the procedure. A significant advantage of the new approach is that realistic multicomponent aerosol models are not required for the retrieval of C. PMID- 18286041 TI - Demonstration of droplet size and vaporization rate measurements in the near field of a two-phase jet with droplet lasing spectroscopy. AB - Droplet lasing spectroscopy has been applied to the measurement of droplet size and evaporation rate in a spray. A single droplet, doped with laser dye, was injected along the centerline of a liquid spray. Filters were used to block the strong elastic-scattering signal. The lasing emission from the doped droplet could be detected against the background with mass loadings of liquid in the spray as high as 20%. An analysis of the spectrum of droplet lasing was used to evaluate the droplet diameter. The evaporation rate of the droplet was obtained from consecutive lasing spectra that were obtained from the same droplet. An error analysis of the drop size and drop evaporation measurements was carried out and showed that accurate measurements of evaporation rates were feasible. PMID- 18286042 TI - Theory and laboratory demonstrations on the use of a nematic liquid-crystal phase modulator for controlled turbulence generation and adaptive optics. AB - We discuss the use of liquid-crystal phase modulators (LCPM's) both as a repeatable disturbance test source and as an adaptive optics corrector. LCPM's have the potential to induce controlled, repeatable, dynamic aberrations into optical systems at low cost, low complexity, and high flexibility. Because they are programmable and can be operated as transmissive elements, they can easily be inserted into the optical path of an adaptive optics system and used to generate a disturbance test source. When used as wave-front correctors they act as a piston-only segmented mirror and have a number of advantages. These include low operating power requirements, relatively low cost, and compact size. Laboratory experiments with a Meadowlark LCPM are presented. We first describe use of the LCPM as a repeatable disturbance generator for testing adaptive optics systems. We then describe a closed-loop adaptive optics system using the LCPM as the wave front corrector. The adaptive optics system includes a Shack-Hartmann wave-front sensor operated with a zonal control algorithm. PMID- 18286043 TI - NO(2) photometer Based on Solid-State Light Sources. AB - A sensitive NO(2) analyzer, that uses a commercial LED emitting near 450 nm, has been developed. The prototype can measure NO(2) concentrations as low as 1 part in 10(9) as defined by the Environmental Protection Agency. The response time of the system is 1 min. In addition, a demonstration of the use of external-cavity frequency-doubled diode laser radiation to accomplish similar measurements is reported. However, because of low nonlinear optical conversion efficiencies, comparable detection limits were not possible. Nonetheless, frequency-doubled diode-laser wavelengths have the potential to reach farther into the UV than LED technology, allowing the detection and measurement of numerous other gaseous species of interest. PMID- 18286044 TI - High-Power As-S Glass Fiber Delivery Instrument for Pulse YAG:Er Laser Radiation. AB - A 3-mum laser-generation delivery instrument that uses chalcogenide fibers with a unique damage threshold and conical radiation input has been developed for medical applications. A new purification and synthesis scheme has been elaborated that yields fibers with a heterophase inclusion content of less than 10(4) cm( 3). In such fibers the damage threshold is 350 J/cm(2) at an average power density of 0.5 kW/cm(2) in a YAG:Er laser operating in the repetitive pulse free running regime with a pulse duration of 350 ms. 1-3 x 10(4) laser pulses were transmitted at a repetition rate of 3 Hz and an average output power of 1 W under the condition of a 15% decrease in the output power. PMID- 18286045 TI - Interferometric optical time-domain reflectometry for distributed optical-fiber sensing. AB - The technique of optical time-domain reflectometry is analyzed to determine the effect of an optical phase modulation on light backscattered in an optical fiber. It is shown that the spatial distribution along the fiber of an external phase modulation can be measured with a spatial resolution close to that of optical time-domain reflectometry. A distributed interferometric sensor arrangement that employs this technique is investigated experimentally, and a satisfactory interrogation of more than 1000 resolution intervals is demonstrated. PMID- 18286046 TI - Mode-field diameter of single-mode optical fiber by far-field scanning. AB - I use the direct far-field method to measure the mode-field diameter of a single mode fiber with an expanded uncertainty of 30 nm, with a coverage factor of 2. For a step-index fiber with a mode-field diameter of approximately 9 mum, the major sources of uncertainty are nonlinearity in the electronics, angular errors and scattered light in the apparatus, and the polarization and noncircularity of the mode of the fiber. The paper concludes by showing an inconsistency in the derivation of the far-field expression for mode-field diameter. PMID- 18286047 TI - Quantitative Raman imaging investigations of mixing phenomena in high-pressure cryogenic jets. AB - A two-dimensional Raman technique was used to investigate mixing phenomena of cryogenic jets under both supercritical and transcritical conditions. The aim of this study was to enlarge the experimental data basis for modeling purposes and to provide quantitative information to help to improve the design of injectors for high-pressure rocket engine combustion chambers. Cryogenic nitrogen, which served as substitute for liquid O(2), was injected into N(2) at room temperature at pressures up to 6.0 MPa. The liquid N(2) jet could be atomized by a coaxial H(2) flow. Raman scattering was generated with a XeF excimer laser. The resulting signal images were discriminated against background by spectral filtering and preferential detection of light with a polarization corresponding to the polarization of the laser, thus making use of the conserved polarization of the Raman-scattered light. The Raman images were converted into density distributions of N(2) and H(2), respectively, as well as into temperature distributions for a variety of experimental conditions. PMID- 18286048 TI - Resonance shift of obliquely illuminated dielectric cylinders: geometrical-optics estimates. AB - Some characteristics of resonant states in obliquely illuminated cylinders are derived from a geometrical-optics point of view. A formula for the resonance shift that is due to tilted illumination is derived and predictions are compared with data from the literature. PMID- 18286049 TI - Feasibility study of synthetic aperture infrared laser radar techniques for imaging of static and moving objects. AB - Techniques for two types of 10-mum band synthetic aperture infrared laser radar using a hypothetical reference point target (RPT) are presented. One is for imaging static objects with a single two-dimensional scanning aperture. Through the simple manipulation of a reference wave phase, a desired image can be obtained merely by the two-dimensional Fourier transformation of the correlator output between the intermediate frequency signals of the reference and object waves. The other, with a one-dimensional aperture array, is for moving objects that pass across the array direction without attitude change. We performed imaging by using a two-dimensional RPT correlation method. We demonstrate the capability of these methods for imaging and evaluate the necessary conditions for signal-to-noise ratio and random phase errors in signal reception through numerical simulations in terms of feasibility. PMID- 18286050 TI - Bubble chamber as a trace chemical detector. AB - A novel concept for trace chemical analysis in liquids has been demonstrated. The technique utilizes light absorption in a superheated liquid. Although a superheated liquid is thermodynamically unstable, a high degree of superheating can be dynamically achieved for a short period of time. During this time the superheated liquid is extremely sensitive to boiling at nucleation sites produced by energy deposition. Observation of bubbles in the superheated liquid in some sense provides amplification of the initial energy deposition. Bubble chambers containing superheated liquids have been used to detect energetic particles; now a bubble chamber is used to detect a trace chemical in superheated liquid propane by observing bubble formation initiated by optical absorption. Crystal violet is used as a test case and can be detected at the subpart-per-10(12) level by using a Nd:YAG laser. The mechanism for bubble formation and ideas for further improvement are discussed. PMID- 18286051 TI - Performance characteristics of soot primary particle size measurements by time resolved laser-induced incandescence. AB - A detailed analysis of various factors that influence the accuracy of time resolved laser-induced incandescence for the determination of primary soot particles is given. As the technique relies on the measurement of the signal ratio at two detection times of the enhanced thermal radiation after an intense laser pulse, guidelines are presented for a suitable choice of detection times to minimize statistical uncertainty. An error analysis is presented for the issues of laser energy absorption, vaporization, heat conduction, and signal detection. Results are shown for a laminar ethene diffusion flame that demonstrate that concurring results are obtained for various laser irradiances, detection characteristics, and times of observation. PMID- 18286052 TI - Pure rotational coherent anti-stokes Raman scattering: comparison of evaluation techniques for determining single-shot simultaneous temperature and relative n(2) o(2) concentration. AB - The accuracy and precision of time-resolved simultaneous temperature and O(2) concentration measurements in binary N(2)-O(2) mixtures by single-pulse dual broadband pure rotational coherent anti-Stokes Raman scattering (CARS) have been investigated. We present a detailed comparison of the applicability of six evaluation procedures to measurements of air in a temperature range 300-2050 K. Special emphasis is put on the dependence of the results on experimental restrictions and distortions. This comparison includes the least-sum-of-squared differences fit (LSF) in the frequency space obtained by use of three different kinds of weighting with respect to signal intensity and in Fourier space by use of the complex or the cosine Fourier transformation, both of which permit a great reduction in the number of data points necessary for multidimensional evaluation. Additionally, a cross-correlation technique is tested that, to the best of our knowledge, was not previously applied to pure rotational CARS. We also present the results of measurements directed to the determination of low O(2) concentration levels that were performed for various binary mixtures (1.0-15.6% O(2)) and for natural air within a temperature range of 300-773 K. A comparison is given for the three evaluation techniques that have proved most promising for the high-temperature investigations, i.e., the constant and the inverse weighted LSF in frequency space and the Fourier analysis technique. PMID- 18286053 TI - Accurate Numerical Analysis of Multimode-Interference-Based 3-dB Couplers. AB - The power coupling characteristics of a multimode-interference-based 3-dB coupler have been analyzed by using the vector finite-element and least-squares boundary residual methods. The numerical results obtained have been compared with published experimental data to show the accuracy and the versatility of these numerical schemes. The results predict that a full treatment of the two dimensional cross section is of particular importance for the design and the simulation of effective waveguide devices to enable a study of their important optical properties such as polarization and fabrication tolerances. PMID- 18286054 TI - Dispersion measurements of water with white-light interferometry. AB - We measure the second- and third-order dispersion coefficients, d(2)k/domega(2) and d(3)k/domega(3), of water for wavelengths from 0.45 to 1.3 mum using a Michelson white-light interferometer. In this interval, the second-order dispersion ranges from 0.068 to -0.1 fs(2)/mum, and the third-order dispersion ranges from 0.048 to 1.18 fs(3)/mum. We observe an oscillation in d(2)k/domega(2) near 1.1 mum that is due to water absorption features near that wavelength. From the dispersion coefficients, derivatives of the index of refraction of water are calculated and compared with available equations. These measured values of d(2)n/dlambda(2) and d(3)n/dlambda(3) should be useful in the evaluation and improvement of existing equations for n(lambda) in water. PMID- 18286055 TI - Signal extraction in a power-recycled michelson interferometer with fabry-perot arm cavities by use of a multiple-carrier frontal modulation scheme. AB - We present a signal extraction scheme for longitudinal sensing and control of an interferometric gravitational-wave detector based on a multiple-frequency heterodyne detection technique. Gravitational-wave detectors use multiple-mirror resonant optical systems where resonance conditions must be satisfied for multiple degrees of freedom that are optically coupled. The multiple-carrier longitudinal-sensing technique provides sensitive signals for all interferometric lengths to be controlled and successfully decouples them. The feasibility of the technique is demonstrated on a tabletop-scale power-recycled Michelson interferometer with Fabry-Perot arm cavities, and the experimentally measured values of the length-sensing signals are in good agreement with theoretical calculations. PMID- 18286056 TI - Energy-Transfer Dye Laser in SolGel Silica. AB - Coumarin 460 transfers radiative energy to disodium fluorescein when solgel silica is doped with the dyes, producing disodium fluorescein lasing in superradiant mode at a 3.3% peak efficiency under low-energy transverse pumping by a 1.5-mJ N(2) laser, with disodium fluorescein being present at a concentration below its lasing threshold value. PMID- 18286057 TI - Analysis of lasing in gas-flow lasers with stable resonators. AB - A model is developed that describes the power extraction in chemical oxygen iodine lasers (COIL's) and CO(2) gasdynamic lasers with stable resonators when a large number of transverse Hermite-Gaussian eigenmodes oscillate. The extraction efficiency, mode intensities, and intensity distribution along the flow depend only on two parameters. The first is the ratio gamma(0) of the residence time of the gas in the resonator to the O(2)((1)D) or N(2)(v) energy extraction time and the second is the ratio of the threshold to the small-signal gain. The efficiency is maximum for gamma(0) ? infinity and decreases rapidly as gamma(0) decreases. It is found that for a range of parameters corresponding to the highest efficiencies the intensity distribution along the flow is nonuniform and has two peaks near the upstream and downstream sections of the resonator. In this case only the highest-order modes that totally fill the resonator cross section oscillate (the so-called, experimentally observed sugar scooping bimodal intensity distribution). For the range of parameters corresponding to smaller efficiencies the intensity is uniform. In this case all the modes participate in lasing; however, the intensities of the high-order modes are larger than those of the low order. The current model is compared with the plane-mirror Fabry-Perot resonator model and with the constant intraresonator intensity and rooftop models of COIL's with stable resonators. The extraction efficiency calculated with the last two models is close to that estimated from our model. However, the intensity distribution cannot be calculated correctly using the Fabry-Perot, the constant intraresonator intensity, or the rooftop model. PMID- 18286058 TI - Injection-seeded, single-frequency, q-switched erbium:glass laser for remote sensing. AB - We have built and characterized an injection-seeded, Q-switched, flash-lamp pumped, eye-safe Er:glass laser that is suitable for coherent remote sensing. The output of the device is a 400-ns, single-frequency, transform-limited pulse of 1 mJ at 1.552 mum. PMID- 18286059 TI - Saturation Parameter and Small-Signal Gain of a Laser-Diode-Pumped Tm:YAG Laser. AB - We have studied the small-signal gain and the saturation parameter of a Tm:YAG laser in a longitudinal-mode oscillation. By using the cavity loss method, a maximum small-signal gain of 4.3%/cm at a crystal temperature of 15 degrees C and a maximum saturation parameter of 13 kW/cm(2) at a crystal temperature of 25 degrees C were determined with an oscillation wavelength of 2013 nm. PMID- 18286060 TI - Efficient 946-nm Laser Operation of a Composite Nd:YAG Rod With Undoped Ends. AB - We present comparative studies on laser performance of diode end-pumped, composite Nd:YAG rods with undoped ends at 946 nm. Efficient heat removal in a composite rod can reduce the peak of rise in temperature in the active segment to 61% by theoretical calculation. The maximum, continuous-wave output power of 1.5 W, which is three times higher than a conventional noncomposite rod, was obtained at an absorbed pump power of 8 W because of the 55 deg reduction in temperature rise that was estimated experimentally. The performance and characteristics of composite rods in quasi-three-level laser operation are analyzed and discussed in detail, taking into account the reduction in reabsorption loss, which is strongly temperature dependent. PMID- 18286061 TI - Two-photon-pumped frequency-upconverted blue lasing in coumarin dye solution. AB - Two-photon-pumped (TPP) frequency-upconverted blue lasing of Coumarin 500 dye solution has been experimentally investigated. The shortest lasing wavelength was measured to be ~479 nm from a Coumarin 500 solution in chloroform pumped with ~800-nm laser pulses of ~5-ns duration. The spectral, temporal, and spatial structures as well as the output-input characteristics of TPP cavity lasing were measured with a 1-cm-long Coumarin 500 solution-filled quartz cuvette. The cavity lasing spectral structure and the numbers of longitudinal modes were easily controlled simply by attachment of an optical plate to the output window of the dye-solution cuvette. The net conversion efficiency from the absorbed 800-nm pump pulse energy to the blue-upconverted cavity lasing energy was ~4.8%. PMID- 18286062 TI - Compact and Efficient 3.2-W Diode-Pumped Nd:YVO(4)/KTP Green Laser. AB - We demonstrate a simple way to achieve single-frequency operation by using fiber coupled diode-pumped Nd:YVO(4)/KTP green lasers in a short standing-wave linear cavity. A single-mode output with 3.2-W green power was generated with a 12.6-W pump power corresponding to a conversion efficiency of 25.4%. The single-mode operation was obtained through the combined action of the anisotropic emission cross section of Nd:YVO(4) and the KTP crystal acting as a birefringent filter. PMID- 18286063 TI - Infrared properties of chemical-vapor deposition polycrystalline diamond windows. AB - Low-resolution transmittance and reflectance spectra of high-quality chemical vapor deposition (CVD) diamond windows were measured in the infrared in the 2.5 500-mum wavelength range (20-4000 cm(-1)). High-resolution measurements on a window with nearly parallel surfaces show well defined interference fringes at low frequencies. By standard procedures the optical constants n and k of CVD diamond were determined, for the first time to the author's knowledge, in the far infrared region. It is shown that a window with a large wedge angle, close to 1 degrees , does not produce appreciable interference fringes. Modeling of these results confirms that interference fringes can be avoided by use of properly wedged CVD diamond windows. This result is of considerable relevance to the use of CVD diamond windows in spectroscopic applications for which fringe suppression is a major requirement. PMID- 18286064 TI - Optical Studies of Nd-doped benzil, a potential luminescent and laser material. AB - Neodymium-doped benzil crystals have been synthesized and characterized for their absorption, emission, and kinetics properties. From Judd-Ofelt analysis, the radiative decay time of Nd emission (peaking at 1055 nm) is estimated to be equal to 441 mus. The experimental Nd lifetime (under Ar+ laser excitation) is equal to 19 mus. The broad emission band centered at approximately 700 nm (tau(decay) approximately 15 ns) and the Raman scattering with characteristic frequency shift of 1600 cm(-1) have been observed at excitation of benzil with 532-nm Q-switched laser pulses. We show that rare-earth-doped benzil can be considered as a potential candidate for luminescent and solid-state laser material. PMID- 18286065 TI - Mid-Infrared-Wavelength Generation in 2-mum Pumped Periodically Poled Lithium Niobate. AB - A periodically poled lithium niobate optical parametric oscillator pumped by a Tm:YAG laser at 2.0124-mum wavelength has been demonstrated. A pump pulse energy of 5.1 mJ generated 0.65 mJ of signal and idler pulse energy at a 50-Hz repetition frequency with a 27.8-mum domain-period-length grating. The lithium niobate crystal at a temperature of 180 degrees C yielded 3.61- and 4.55-mum signal and idler wavelengths, respectively. Wavelength tuning over a wide range was achieved with domain-period lengths from 25.5 to 28.2 mum and crystal temperature tuning from 50 to 180 degrees C. Signal wavelengths of 3.26-3.76 mum and idler wavelengths of 4.33-5.34 mum were generated. PMID- 18286066 TI - Finite-Element Analysis of Lossy TE-TM modes in Metal-Clad Optical Waveguides. AB - Finite-element analysis employing the scalar and vector H-field formulations and with the aid of the perturbation technique is used to calculate the TE-TM complex propagation characteristics of integrated optical devices in gallium arsenide, lithium niobate, and silica fiber, incorporating a lossy metal cladding. The propagation and attenuation properties of several types of metal-clad planar optical waveguide, which exhibit surface-plasmon properties for the TM polarization, are reviewed, and the modal loss caused by the metal cladding in a titanium-diffused lithium niobate electro-optic directional coupler modulator, an indium gallium arsenide phosphide-based TE-TM optical polarizer, and a submicron metal-clad silica fiber suitable for near-field optical scanning microscopy is calculated. PMID- 18286067 TI - Fiber-coupled high-power external-cavity semiconductor lasers for real-time Raman sensing. AB - High-power, external-cavity semiconductor lasers with narrow bandwidth and fiber coupled output are designed and constructed. An output power of 540 mW is coupled out of a 100-mum multimode fiber with coupling efficiency of 72% when the laser is operated at 1.1 A. The emission linewidth is as narrow as 22 GHz, and the wavelength is tunable from 779.7 to 793.0 nm. Application of such lasers to remote real-time Raman sensing of materials is also demonstrated. PMID- 18286068 TI - Imaging spectroscopy for two-dimensional characterization of auroral emissions. AB - A large throughput transmission spectrometer, with a grating on a prism as the diffraction element, has been developed to study altitude distributions of auroral emissions. The imaging spectrometer disperses spectrally in one dimension while spatial information is preserved in the orthogonal direction. The image is projected onto a CCD array detector. Image processing methods have been developed to calibrate for wavelength, uniform field, spectral sensitivity, curvature of field, and spatial mapping. Single images are processed to represent a measured signal brightness in a unit of Rayleighs/pixel, from which area integrations can be made for desired spatial-spectral resolution. System performance is ~1.5-nm resolution over a 450-nm bandwidth (420-870 nm). Two spectrometer systems of this design were operated simultaneously, one with additional optical instruments and an incoherent scatter radar at Sondrestrom, Greenland, and the other at Godhavn, Greenland, which lies 290 km to the northwest and nearly in the magnetic meridian of Sondrestrom. The developed system, calibration method, and examples of performance results are presented. PMID- 18286069 TI - High-sensitivity mid-infrared heterodyne spectrometer with a tunable diode laser as a local oscillator. AB - A new mid-IR heterodyne spectrometer, which is intended to be applied for atmospheric and astrophysical studies, is presented. The spectrometer uses a frequency-stabilized tunable diode laser as a local oscillator. Owing to the low output power of available single-mode diode lasers, a newly developed confocal ring resonator, the diplexer, is used to superimpose the source signal efficiently with that of the local oscillator. Additionally, the diplexer serves as an optical filter that establishes controlled optical feedback between the laser diode and the detector, which allows stable laser operation with linewidths of the order of 1 MHz. The heterodyne signal from the HgCdTe detector is analyzed by means of a 1.4-GHz acousto-optical spectrometer. With this setup we find system temperatures as low as 4400 K (double sideband), that is, approximately a factor of 6 of the quantum limit. PMID- 18286070 TI - Dual-purpose, compact spectrometer and fiber-coupled laser wavemeter based on a wollaston prism. AB - A fiber-coupled, compact laser wavemeter based on a modified Wollaston prism has been constructed and evaluated. The path difference between orthogonal polarization states of the input light varies smoothly across the aperture of the prism forming an interferogram in the spatial domain that is recorded with a CCD detector array. A Fourier transform of this interferogram gives the spectral distribution of the incident light. Alternatively, for a narrow-linewidth source a fringe period measurement technique is used to obtain precision measurement of the center wavelength. Using 752 interferogram data points we obtain a wavelength precision of 1 part in 10(6). The elimination of moving parts from the design makes the recorded interferogram inherently stable. PMID- 18286071 TI - Patents. PMID- 18286072 TI - Atmospheric Absorption in Laboratory Measurements: A Comparison Between modtran3 and Measurement. AB - Errors can occur in laboratory measurements when the response of a bandpass filtered radiometer extends into an atmospheric absorption region. Atmospheric models, such as modtran3, can be valuable tools that permit the optical measurements in these regions to be accurately analyzed, provided the models themselves are accurate. Comparisons of modtran3-predicted and laboratory measured atmospheric transmittance have been made to help establish the validity of modtran3 for use in modeling short-path-length (a few meters or less), low resolution optical effects over a wavelength range of 700-5700 nm. Comparisons include percentage differences, range of differences, and band-averaged differences. Good agreement is shown for all absorption bands except for the CO(2) band near 4300 nm. In general, the band-average differences are less than ~0.75% for all water bands. For the CO(2) band the modtran3 default CO(2) level is found to be inappropriate, and 610 parts per million in volume is found to give much better agreement with measurement. PMID- 18286073 TI - Oblique incidence of gaussian beams on waveguide gratings with the four-wave coupled-mode theory. AB - Diffraction of obliquely incident Gaussian beams on waveguide gratings with infinitely long grooves is described. The analysis is based on the angular spectrum of the beam and on the grating response derived from the four-wave coupled-mode theory, which simultaneously considers all four (approximately) synchronous waves. Results concerning the shapes of the emerging beams and their directions, the power content, and the possibility of beam splitting could differ significantly from those obtained by means of two-wave coupling. The analysis method is general and can be performed on many kinds of realistic beam shapes and applications. PMID- 18286074 TI - Numerical and experimental study of grating efficiency for synchrotron monochromators. AB - The development of third-generation synchrotron sources has stimulated efforts toward high-resolution monochromators. A good knowledge of grating efficiency is needed to achieve an optimal compromise between resolution and photon flux. Because simple geometric models fail to describe correctly the gratings properties in the UVtosoft-X-ray range, we have developed a simulation software based on differential theory. A simplified R-matrix propagation algorithm assures the numerical stability of the code for deep gratings. Our numerical results are compared with previous research on deep gratings. Experimental and numerical studies have been performed on some test cases at a synchrotron source. Very good agreement between numerical prediction and measurement has been found. PMID- 18286075 TI - Rectangular surface-relief transmission gratings with a very large first-order diffraction efficiency (~95%) for unpolarized light. AB - Computer optimization shows that the first-order diffraction efficiency of a lossless-transmission surface-relief grating with a rectangular surface profile can be made very large (~95%) simultaneously for light of TE and TM polarizations incident near the Bragg angle by the proper choice of the fill factor. The case for visible light incident close to the Bragg angle on unslanted gratings with periodicities corresponding to Bragg angles of 30 degrees , 37.5 degrees , and 45 degrees is presented. The refractive index of the grating material was chosen in the range between 1.2 and 2. PMID- 18286076 TI - Optical scatterometry of subwavelength diffraction gratings: neural-network approach. AB - Optical scatterometry is a method for the on-line measurement of the geometry of a diffraction grating, which is deduced from diffraction-pattern data. We demonstrate the use of a neural network as a promising method for performing an accurate quantitative characterization of the geometry. As an example, we show the deduction of the geometry of a grating with subwavelength grooves with a rms accuracy of 1.9 degrees for the slope of the groove walls, 0.7 nm for the linewidth, and 1.0 nm for the groove depth. PMID- 18286077 TI - Evaluation of diffraction loss in prism light guides by finite-difference time domain field modeling. AB - Prism light guides are hollow dielectric tubes that use prismatic facets to guide light by means of total internal reflection. An unresolved problem has been to determine the magnitude of loss caused by diffraction in prism light guides. Neither experimental measurement nor an analytical solution has yet been achieved, so we attacked the problem numerically, in two steps. First, we found a way to represent such a transitionally invariant three-dimensional system as an equivalent two-dimensional problem. Second, we employed the finite-difference time-domain algorithm, with periodic boundary conditions, to yield a computation problem of manageable size. We found that the diffraction-induced transmissivity of a prism light guide wall is of the order of the wavelength divided by the prism size-a result that has encouraging practical implications. PMID- 18286078 TI - Rigorous coupled-wave analysis for practical planar dielectric gratings: 1. Thickness-changed holograms and some characteristics of diffraction efficiency. AB - A new and useful method for obtaining diffraction efficiencies from holograms manufactured practically is presented. Applying the rigorous coupled-wave analysis, we express each difference between the practical and the ideal as a mathematical component that can be easily integrated. In Part 1 the effects due to thickness change in the hologram layer (observed frequently after the development process) are treated. Although uniform swelling or shrinking causes a simple reconstruction wavelength or incidence-angle shift, nonuniform thickness extends the capacity of the Bragg condition matching, creating a diffraction efficiency curve in the asymmetric profile. Other characteristics of diffraction are also maintained. A refractive-index change has an effect that is similar to the thickness change. Higher-order terms in permittivity modulation create negligible effects in general holograms when used at or near the simple first order Bragg condition. PMID- 18286079 TI - Rigorous coupled-wave analysis for practical planar dielectric gratings: 2. Diffraction by a surface-eroded hologram layer. AB - The diffraction efficiencies of practical dielectric holograms are evaluated with rigorous coupled-wave analysis. The cases of the hologram surfaces eroded in several shapes are treated and compared with those in which the surfaces are not eroded and with those in which there are only surface gratings. Eroding the surface will increase the higher-order reflection diffraction efficiencies and the transmissions, thus reducing the first reflection and the zeroth transmission. However, sealing the hologram with a cover plate, as is done in manufacturing many holograms, extinguishes the erosion effect. PMID- 18286080 TI - Rigorous coupled-wave analysis for practical planar dielectric gratings: 3. Increase of higher-order lights owing to degenerated complex diffraction. AB - The diffraction efficiencies by practical dielectric holograms are evaluated with the rigorous coupled-wave analysis. As discussed in Parts 1 and 2, only the first reflection light can be diffracted by general sealed volume reflection holograms. Here we discuss the possibility that complex diffraction, which occurs in multiple-grating-storage holograms, makes diffraction efficiencies of higher order lights increase drastically, i.e., the degenerated complex diffraction. Although evident in special cases only, they appear frequently in actual holographic use. PMID- 18286081 TI - Influence of several grating sensor parameters on the modulation depth of moire signals under incoherent illumination. AB - Theoretical expressions for the modulation depth of moire signals under incoherent illumination are derived. Consequently, the modulation depth of a moire signal is related to the following factors: the geometric shape and the size of the light source; the diffraction effect of the grating, which relates to the number of lines in the grating; the line and the space ratio; the grating pair gap; the geometric shape and the size of the receiving window; etc. In addition, the influence of the grating pair on the period and the inclination of moire fringes under noncollimated illumination are discussed, and the changes in the moire signal modulation depth under noncollimated illumination with that under collimated illumination are made. Finally, some experimental results are given to verify the theoretical expressions. This research is useful for the actual design of grating sensors. PMID- 18286082 TI - Broadband and tuned signal recycling with a simple michelson interferometer. AB - We present experimental data on the frequency response of both broadband and tuned signal recycling with a benchtop Michelson interferometer. These data are in excellent agreement with our simple theoretical model. We use in-line modulation to give a control system that provides a high degree of orthogonality between the two servo loops. PMID- 18286083 TI - Automatic processing in moire deflectometry by local fringe direction calculation. AB - An algorithm for accurately extracting the local fringe direction is presented. The algorithm estimates, in the neighborhood of n x n points, the direction of the gradient that points normal to the local fringe direction. The performance of four different derivative kernels is also compared. Since this method is sensitive to noise and variations in background and amplitude, a preprocessing step is used to limit these error sources. The method has been applied to the moire deflectogram of a spherical and a progressive addition ophthalmic lens, resulting in a map of the refractive power of these lenses. The results are compared with the data obtained with a commercial focimeter. This technique is useful for analyzing the fringe patterns where the fringe direction is variable and must be obtained locally. PMID- 18286084 TI - Spectroscopic ellipsometry of slightly inhomogeneous nonabsorbing thin films with arbitrary refractive-index profiles: theoretical study. AB - We develop a new approximation for the amplitude reflection coefficients of a slightly inhomogeneous thin film. This approximation incorporates exactly the interference effects at the substrate and the ambient interfaces. Interference effects inside the inhomogeneous film are incorporated in the Born approximation. We also develop a new approach to the reconstruction of the refractive-index profile from ellipsometric spectra. It is based on a physically sound parameterization of the refractive-index profile. The new approach is tested on the model reconstruction problem. PMID- 18286085 TI - Standards for Which the Ellipsometric Parameter psi Remains Insensitive to Variations in the Angle of Incidence. AB - We examine the possibility of using two- and three-phase systems suitable as standards for which the ellipsometric parameter ? remains insensitive to variations in the angle of incidence. These standards avoid propagation of errors in the angle of incidence with respect to the measured standard ? value. Different materials (dielectrics, metals, and semiconductors), adequate for the above purpose, are considered in different structure combinations, and their optical response are analyzed. PMID- 18286086 TI - Fourier-transform radiation thermometry: measurements and uncertainties. AB - A Fourier-transform radiometer is used to measure blackbody temperatures in the 5001000-K range. The measurements involve collecting mid-infrared spectra at two known reference temperatures and one unknown temperature. The accuracy of the interpolation measurement technique is discussed, and the effects of the uncertainty in the temperature reference points, the voltage ratio measurement, and the wavelength accuracy are described. Temperature accuracy at the 0.5% level has been achieved; the main uncertainty component is caused by the interferometer drift. Directions to reach 100-mk accuracy levels have been identified. PMID- 18286087 TI - Laser Doppler velocimetry with a compact disc pickup. AB - A laser Doppler velocimeter employing a compact disc pickup for both fringe projection and signal detection is described. The spectrum of the recorded signal gives the information about the speed of the object. The device takes advantage of the Talbot effect to project the grating contained in the pickup onto a moving target, so that no imaging system is required. The peculiar imaging technique allows for the exploitation of several optical configurations and permits the manipulation of the intensity profile of the projected grating. The instrument was used to measure the velocity of dust particles on a solid substrate in the 1 m/s range but could also find an application to the study of liquid flow. PMID- 18286088 TI - Broadband Division-of-Amplitude Polarimeter Based on Uncoated Prisms. AB - A broadband division-of-amplitude polarimeter (DOAP) is presented. It can provide the real-time measurement of any state of polarization of light, described by its Stokes vector, in large spectral windows. The light is split first into two beams by a prism and then into four beams by means of any polarizer device that will separate the two linear orthogonal states of polarization. Finally, the Stokes vector is directly deduced from the four measured intensities. To avoid interference effects, the splitting of light into four beams is induced only by refractive-index contrast effects between semi-infinite media that are weakly dependent on the wavelength. An experimental setup working from 0.4 to 2 mum is described. It provides similar sensitivities for all the states of polarization, and its characteristics are constant, on a scale of a few percent, within the spectral window. Calibrations performed at 458 and 633 nm display good agreement between theoretical and experimental values. The accuracy of the prism DOAP, evaluated by measurement of the Stokes vector produced by a rotating Glan polarizer, is better than 1%. An infrared extension of this polarimeter is also presented. PMID- 18286089 TI - Fiber-optic pulsed photothermal radiometry for fast surface-temperature measurements. AB - Temperature measurement based on pulsed photothermal radiometry is described. In this technique a body is irradiated by a laser pulse and its temperature is inferred from the shape of the emitted photothermal-signal curve. A prototypical system based on a pulsed CO(2) laser, an IR detector, and IR-transmitting silver halide optical fibers was constructed and used to evaluate the feasibility of this technique. An important feature of the technique is that changes in sample emissivity or geometric factors do not introduce errors in the temperature determination. Theory, simulation, and experimental results are given and discussed. PMID- 18286090 TI - Pixel-based absolute topography test for three flats. AB - We demonstrate a method of performing the absolute three-flat test by using reflection symmetries of the surfaces and an algorithm for generating the rotation of arrays of pixel data. Most of the operations involve left/right and top/bottom flips of data arrays, operations that are very fast on most frame grabbers and are available on most commercial phase-measuring interferometers. We demonstrate the method with simulated data as well as with actual data from 150 mm-diameter surfaces that are flat to less than 25 nm peak to valley. PMID- 18286091 TI - Interferometer for small-angle measurement based on total internal reflection. AB - We describe a new method for angle measurement based on the internal-reflection effect and heterodyne interferometry. A novel prism assembly is designed that can always parallel retroreflect the incoming light beams so the optical configuration is compact. As a differential common-path optical configuration is integrated into the design, the linearity of the method is greatly improved. Details of theoretical analysis of the method and experimental verification of the principle are presented. The resolution can be better than 0.3 arc sec. The experimental results and further improvements of the proposed method are also addressed. PMID- 18286092 TI - Absolute Refractive Indices and Thermal Coefficients of Fused Silica and Calcium Fluoride Near 193 nm. AB - The refractive indices of several fused silica and calcium fluoride samples from different suppliers were measured with the minimum deviation method in the deep UV between 191 and 196 nm with a standard uncertainty of 7 ppm. For both materials the dispersion dn/dlambda near 193 nm and 20 degrees C was determined. In addition, we measured the thermal coefficients of the refractive index near 193 nm and between 15 and 25 degrees C. PMID- 18286093 TI - Testing aspheric surfaces: simple method with a circular stop. AB - As an extension of the knife-edge test, a noninterferometric method for inspecting circularly symmetric aspheres is proposed in which the test surface is illuminated by a spherical wave. When a small circular stop is placed around the curvature center of the best-fitting sphere, only rays characteristic for the deviation from a perfect sphere remain unobstructed. An image of the tested surface is thereby obtained, showing light and dark regions whose boundaries are correlated to the surface profile, the stop size, and the stop position along the symmetry axis. The experiment has been carried out with a paraboloid. PMID- 18286094 TI - Optical properties (bidirectional reflection distribution functions) of velvet. AB - A detailed investigation has been made of the unusual characteristics of the angular distribution of surface scattering from velvet in the visual region. We present a novel method in which samples of velvet fabric are wrapped around a right-circular cylinder so that reemitted radiance can be measured by a digital CCD camera. This setup makes it relatively simple to acquire a large set of bidirectional reflection distribution function (BRDF) samples. The study reveals that, apart from the grazing specular lobe and an anisotropic backscattering peak near 50 degrees , the overall BRDF's are rather uniform across the whole angular span of observation. Attempts are made to relate these scattering characteristics to the physical and the geometrical structure of velvet. PMID- 18286095 TI - Backscatter linear and circular polarization analysis of roughened aluminum. AB - A study of cross-polarized and copolarized intensities backscattered from roughened aluminum surfaces is presented for both linear and circular incident polarization states. The angular variation of measured Mueller matrices is shown to contain only diagonal elements, as predicted by the reciprocity theorem. The ratio of cross-depolarized to copolarized scattered intensities is significantly larger for circular than for linear input polarization states. In the linear case the ratio saturates beyond 50 degrees , whereas in the circular case the ratio continues to increase monotonically with angle. A phenomenological model for copolarization and cross-polarization intensities is shown to predict the observed behavior of both linear and circular input polarization states up to incident angles of 70 degrees . PMID- 18286096 TI - Optical constants of sputter-deposited ti-ce oxide and zr-ce oxide films. AB - Films of Ti oxide, Zr oxide, Ce oxide, Ti-Ce oxide, and Zr-Ce oxide were made by means of reactive dc magnetron sputtering in a multitarget arrangement. The films were characterized by x-ray diffraction and electrochemical measurements, both techniques being firmly connected to stoichiometric information. The optical constants n and k were evalued from spectrophotometry and from variable-angle spectroscopic ellipsometry. The two analyses gave consistent results. It was found that n for the mixed-oxide films varied smoothly between the values for the pure oxides, whereas k in the band-gap range showed characteristic differences between Ti-Ce oxide and Zr-Ce oxide. It is speculated that this difference is associated with structural effects. PMID- 18286097 TI - Carbon/Titanium multilayers as soft-x-ray mirrors for the water window. AB - C/Ti multilayers with a period thickness of 2.1-2.7 nm were produced by electron beam evaporation in ultrahigh vacuum as soft-x-ray mirrors in the water window (lambda = 2.3-4.4 nm). For smoothing the individual interfaces and thus enhancing the total reflectance, each layer was ion polished with an Ar(+) ion beam after deposition. For a multilayer of 85 bilayers, a reflectance of approximately 11% at an angle of incidence of 59 degrees (with respect to the surface normal) by use of s-polarized radiation at a wavelength of 2.77 nm was achieved. PMID- 18286098 TI - Ice-Front Propagation Monitoring in Tissue by the use of Visible-Light Spectroscopy. AB - For demonstrating that visible-light spectroscopy can be used for ice-front detection within freezing tissue, proton magnetic resonance images were correlated to time-evolving transmittance spectra as an ice front progressed across a tissue sample. The experimental apparatus was designed to be compatible with magnetic resonance imaging, to produce one-dimensional freezing, and to allow both reflectance and transillumination emitter-detector configurations about a normally progressing planar ice front in chicken muscle. This demonstration has potentially important medical applications in cryopreservation (freezing of biological materials for preservation) and cryosurgery (destruction of tissue by freezing). PMID- 18286099 TI - Optoelectronic region of interest detection: an application in automated cytology. AB - Diagnostic cytology, which is used to screen for cervical cancer, involves characterizing cellular features such as shape, size, and texture. Automated screening of cervical smear slides is desirable but computationally challenging since each slide requires processing 2 x 10(9) pixels at a resolution of 0.8 mum per pixel. We demonstrate that the throughput of optical processors can be exploited in automated cervical smear-screening systems. In particular, we identify a morphological shape detector to perform the initial region of interest (ROI) detection and to demonstrate experimentally its optoelectronic implementation. The ROI detector is tested on 200 images, and its performance is characterized as a receiver operating characteristic (ROC). The area under the ROC curve is as high as 96.4% of the total area. The simulation and the experimental results are found comparable, and the discrepancy between the two results is determined to be a function of the number of bits represented in the filter plane device. PMID- 18286100 TI - Mapping of Birefringence and Thermal Damage in Tissue by use of Polarization Sensitive Optical Coherence Tomography. AB - We demonstrate cross-sectional birefringence- and polarization-independent backscatter imaging of laser-induced thermal damage in porcine myocardium in vitro, using a polarization-sensitive optical coherence tomography system. We compare the generated images with histological sections of the tissue and demonstrate that birefringence is a more sensitive indicator of thermal damage than is backscattered light. Loss of birefringence in thermally damaged regions is quantified and shown to have significant contrast with undamaged sections of the tissue. A detailed theoretical analysis of the birefringence measurements is provided, including a calculation of the systematic errors associated with background noise, system imperfections, and tissue dichroism. PMID- 18286101 TI - Distinguishability of biological material by use of ultraviolet multispectral fluorescence. AB - Recent interest in the detection and analysis of biological samples by spectroscopic methods has led to questions concerning the degree of distinguishability and biological variability of the UV fluorescent spectra from such complex samples. We show that the degree of distinguishability of such spectra is readily determined numerically. As a practical example of this technique, we show its application to the analysis of UV fluorescence spectra taken of E. coli, S. aureus, and S. typhimurium. The use of this analysis to determine the degree of biological variability and also to verify that measurements are being made in a linear regime in which analytic methods such as multivariate analysis are valid is discussed. PMID- 18286102 TI - Computer-aided design for optoelectronic systems: introduction to the feature issue. AB - Welcome to the first special issue of Applied Optics on computer-aided design for optoelectronic systems. This special issue stemmed from our realization of the need for dialogue between optoelectronic system designers and computer-aided design developers, as well as from the realization that various research groups are developing or, in some instances, have already developed and commercialized such tools. Our goal for this special issue is to enhance this type of dialogue by showing to the optoelectronic system design community the current state of optoelectronic computer-aided-design tools. PMID- 18286103 TI - Communication Systems Interactive Software (COMSIS): Modeling of Components and its Application to the Simulation of Optical Communication Systems. AB - comsis, which stands for communication systems interactive software, is a computer-aided-design tool based on a time approach. It allows the design, analysis, and performance optimization of optical transmission systems by use of various optical devices. comsis allows scalar or vectorial simulations, depending on whether the polarization is taken into account. An overview concerning the optical component models is given. A new model of an erbium-doped fiber amplifier allows the user to describe the amplifier through either physical or system parameters by using silicate or fluoride glass fibers or any other material, provided the user can give a file that contains the amplifier's characteristics. The new model of a single-mode fiber allows the user to describe chromatic dispersion through a constant, a function, or a file (given by the user) and to take optionally into account the Kerr and the Raman effects and the polarization mode dispersion. The simulation tools that are used to characterize the quality of an optical transmission system are also presented. To show the system's full range of capabilities in the optical domain, we describe examples of wavelength division-multiplexing and soliton-transmission systems. PMID- 18286104 TI - Two simulators for photonic computer-aided design. AB - The computer-aided design of photonic systems offers many advantages for systems design, optimization, and planning. We present a comparison of two simulation packages for photonic-device, circuit, and system design. Both were developed by the Australian Photonics Cooperative Research Centre: the optoelectronic, photonic, and advanced laser simulator performs step-by-step simulations on complex photonic devices, circuits, and systems, allowing for backward waves and compound resonators; the gigabit optical link designer uses block processing of periodic waveforms to give an efficient estimation of eye diagrams and bit-error rates of time-division-multiplexed and wavelength-division-multiplexed systems. The gigabit optical link designer and the optoelectronic, photonic advanced laser simulator are compared, and examples of typical applications are given. PMID- 18286105 TI - Chatoyant: a computer-aided- design tool for free-space optoelectronic systems. AB - Chatoyant is a tool for the simulation and the analysis of heterogeneous free space optoelectronic architectures. It is capable of modeling digital and analog electronic and optical signal propagation with mechanical tolerancing at the system level. We present models for a variety of optoelectronic devices and results that demonstrate the system's ability to predict the effects of various component parameters, such as detector geometry, and system parameters, such as alignment tolerances, on system-performance measures, such as the bit-error rate. PMID- 18286106 TI - SPICE-Based Optoelectronic System Simulation. AB - spiceis a widely used simulation tool for electrical circuits and systems. When optoelectronic elements, such as lasers, detectors, modulators, etc., and optical elements, such as lenses, gratings, beam splitters, etc., are incorporated into spice, optoelectronic system simulation can be combined with that of electronic systems, facilitating hybrid system design and analysis. We discuss an optoelectronic spice implementation that includes time-, power-, and wavelength domain behaviors. Examples of optical component simulation and optical interconnect system simulation by use of spice are included and compared with the results from a conventional ray-tracing optical analysis tool. PMID- 18286107 TI - Hardware Description Language for Optical Processing (hadlop): a Simulation Environment for Parallel Optoelectronic Architectures. AB - We present a simulation environment for parallel optoelectronic data-processing systems, and we especially consider the fusion of optoelectronic integrated circuits and optical interconnection modules. hadlop, which stands for hardware description language for optical processing, is a simulator that works at the digital design level. So far, hadlop has allowed algorithm and architecture studies for smart-pixel systems. We have just begun to extend the capabilities of hadlop toward an automatic synthesis tool for three-dimensional optoelectronic VLSI circuits. A hadlop architecture will then be the basis for the automatic generation of detailed construction plans that consider the interaction between optical interconnection modules and optoelectronic integrated circuits. The simulation system is freeware and is available through the Internet at http://www2.informatik.uni-jena.de/pope/HADLOP/hadlop.html. PMID- 18286108 TI - Optical Storage Interactive Simulator (oasis): An Interactive Tool for the Analysis of Page-Oriented Optical Memories. AB - Page-oriented optical memories combine high capacity with massive data-transfer rates and could provide the next generation of secondary storage systems. Several technological barriers need to be overcome before these memories become commercial products. One is the need for efficient interfaces to electronic computers. To assist in the analysis of all issues pertaining to such an interface, we have developed the optical storage interactive simulator (oasis), an interactive software tool. oasis can simulate data-encoding schemes, noise sources present in a particular memory system, and data-recovery mechanisms. Bit error rates and other useful statistics can also be measured. The user has the option of studying the effects of individual error sources to the system output or of applying several of them in any order dictated by the system architecture. This multifaceted analysis will assist the user in evaluating the effectiveness of a particular error-correcting code and choosing the right one for the system. PMID- 18286109 TI - Simulating free-space optical computing architectures. AB - Major issues in optoelectronic system design include timing, synchronization, and control. Designing free-space optical computing architectures is difficult because of the high degree of system complexity, parallelism, and concurrency in conjunction with the high cost and lack of availability of devices. Current simulation tools lack the expressiveness to model the system structure and behavior of parallel and concurrent architectures, thus making them inefficient and ineffective. We show that Petri nets, compared with other system-modeling methodologies, are more efficient and effective at expressing the functional, behavioral, and structural properties of parallel and concurrent architectures. We show how an extended version of the standard Petri net, a timed-colored Petri net, is used to model and simulate free-space optoelectronic computing architectures. We also present methods for analysis of system timing, synchronization, and control behavior. PMID- 18286110 TI - Small-signal-equivalent circuits for a semiconductor laser. AB - Passive electrical circuits whose voltage and current equations are exactly equivalent to the small-signal rate equations of a semiconductor laser are derived to model an electrically modulated laser (verified to be the same as that given in the literature), an optically modulated laser (i.e., a laser used as an optical amplifier), and a multimode laser. These circuits offer a fast and efficient simulation tool with little computational complexity in which the small signal assumption (i.e., small modulation range) is neither violated nor insufficient for the simulation. PMID- 18286111 TI - Automated design of integrated circuits with area-distributed input-output pads. AB - We present a method for automating the creation of complementary-metal-oxide semiconductor (CMOS) integrated circuits that successfully utilizes a large number of area-distributed pads for input-output communication. This method uses Duet Technologies' epoch computer-aided-design tool for automated placement and routing of CMOS circuitry, given a schematic netlist as an input. The novelty of this approach is that it uses Duet Technologies' eggo program to place and route area-pad signals. To verify this methodology, it is applied to the design of a digital signal-processing circuit, with 200 optical area-pad input-outputs and 44 perimeter-pad input-outputs, that is being fabricated with Bell Labs 1997 CMOS multiple-quantum-well foundry. The layout results are as good as or better than the results obtained by manual layout. PMID- 18286112 TI - Cost, performance, and reliability simulator for optical transceiver modules. AB - The program couple combines simulators for optical performance, mechanical reliability, and production cost under a graphical-user interface to design, simulate, and evaluate micro-optomechanical structures. The thermal simulator predicts the package temperature distribution on the basis of the materials and the geometry as well as on heat sources, sinks, and boundary conditions. The thermal distribution is input to the mechanical simulator, which calculates the stresses or strains and displacements caused by differential thermal expansion. The optical simulator predicts the impact on the optical efficiency and the cross talk of mechanical and optical parameter variations such as solder heights, misalignments, and wavelength distributions. The cost simulator represents the manufacturing process flow and calculates the final cost and the cost sensitivity on basis of the cost and the yield of each process step. By means of balancing detector and coupling yield, cosimulation from optical to cost domains determines the optimum detector size to produce the lowest-cost transceiver module. PMID- 18286113 TI - Photoinduction of anisotropic grains in organic compounds. AB - We report the linear polarized light-induced anisotropic grains in azo-dye colored gelatin films. The optical axes of the grains (fast or slow) are directed along the polarization vector of the exciting light. PMID- 18286114 TI - Achromatic fourier transforming properties of a separated diffractive lens doublet: theory and experiment. AB - The strong chromatic distortion associated with diffractive optical elements is fully exploited to achieve an achromatic optical Fourier transformation under broadband point-source illumination by means of an air-spaced diffractive lens doublet. An analysis of the system is carried out by use of the Fresnel diffraction theory, and the residual secondary spectrum (both axial and transversal) is evaluated. We recognize that the proposed optical architecture allows us to tune the scale factor of the achromatic Fraunhofer diffraction pattern of the input by simply moving the diffracting screen along the optical axis of the system. The performance of our proposed optical setup is verified by several laboratory results. PMID- 18286115 TI - Design of diffractive optical elements for multiple wavelengths. AB - A method for producing diffractive optical elements (DOE's) for multiple wavelengths without chromatic aberration is described. These DOE's can be designed for any distinct wavelength. The DOE's are produced from two different optical materials, taking advantage of their different refractive indices and dispersions. PMID- 18286116 TI - Photorefractive Beam splitter for Free-Space Optical Interconnections. AB - A photorefractive beam splitter (PRBS) is introduced as an alternative to a polarizing beam splitter (PBS) for coupling optical power into reflective modulators in a free-space optical interconnection system. The PRBS uses a single diffraction grating recorded in a photorefractive material to redirect the incident laser light into the first diffraction order and onto the modulators. Reflected interconnection light not matching the Bragg angle criteria transmits uncoupled through the beam splitter. Experimental results show that the PRBS provides better, more uniform transmission for off-axis beams than the currently used PBS. PMID- 18286117 TI - Three-dimensional large-screen display with reflection-mode spatial light modulators and a single-projection optical system: analysis of a retardation modulation method. AB - A stereoscopic projector with polarized glasses is proposed that consists of spatial light modulators (SLM's) that control the retardation of projected light, a polarization beam splitter (PBS), and single-projection optics. This display's features include a three-dimensional (3-D) image display with a single projector and half the size and the power consumption of a conventional 3-D projector. Analysis shows that the cross talk and the extinction ratio of this system depend strongly on the polarized light-separation characteristics of the PBS, the light output, and the extinction ratio of the SLM's. A double-PBS method that drastically improves 3-D image quality is also discussed. PMID- 18286118 TI - Optimal cascade operation of optical phased-array beam deflectors. AB - An optimal strategy for cascading phased-array deflectors is presented that allows for high-resolution random-access beam steering with continuous scan-angle control but requires a minimum number of control lines. The system is analyzed theoretically by use of a Fourier optics approach and then verified experimentally. A pair of 32-channel optical phased arrays fabricated by use of surface electrodes on lanthanum-modified lead zirconate titanate (PLZT) was sandwiched together to form a functional two-stage phased-array cascade. Experimental results from the PLZT-based two-stage deflector are presented that confirm the performance enhancements of the optimized cascading technique. A phase-staggered discrete-offset-bias protocol for controlling the cascaded system is shown to be optimal in terms of maximum diffraction efficiency and minimum number of control lines, while still providing for full analog scan control. PMID- 18286119 TI - Discrete cosine transform domain restoration of defocused images. AB - In discrete-cosine-transform-based (DCT-based) compressions such as JPEG it is a common practice to use the same quantization matrix for both encoding and decoding. However, this need not be the case, and the flexibility of designing different matrices for encoding and decoding allows us to perform image restoration in the DCT domain. This is especially useful when we have severe limitations on the computational power, for instance, with in-camera image manipulation for programmable digital cameras. We provide an algorithm that compensates partially for a defocus error in image acquisition, and experimental results show that the restored image is closer to the in-focus image than is the defocused image. PMID- 18286120 TI - Phase Retrieval From Fresnel Zone Intensity Measurements by use of Gaussian Filtering. AB - The reconstruction of a complex-valued object from intensity measurements in the Fresnel zone region is considered. The reconstruction is based on the phase retrieval method from Fresnel zone intensity data obtained by modulation of the object with a known Gaussian function and with its shifted functions along the horizontal and the vertical directions in rectangular coordinates. Two types of reconstruction system are presented. In the two systems a Gaussian amplitude filter and a Gaussian illuminating beam are used as the known Gaussian functions. Computer-simulated examples in two dimensions illustrate the performance of the reconstruction in these systems. PMID- 18286121 TI - Improved phase-shifting method for automatic processing of moire deflectograms. AB - An improved moire deflectometry phase-shifting technique is presented. A squared grating is used to multiplex the information of the deflections in two orthogonal directions in one image. This procedure avoids the need to rotate the gratings to obtain complete deflection information. However, the use of these gratings makes impossible the application of standard phase-shifting algorithms. Specifically, the problems associated with the nonsinusoidal profile of the moire fringes and the low-modulation areas produced by the square gratings are solved. A modified moire deflectometry phase-shifting method is designed to deal with these problems. In addition, a method to obtain the zero order of the prismatic effect is developed. The technique configures a complete and automatic method of mapping ray deflections. From them the refractive power maps can be derived. Experimental results obtained with a progressive-addition lens are shown. PMID- 18286122 TI - Estimation of the surface velocity of debris flow with computer-based spatial filtering. AB - A computer-based spatial-filtering velocimeter to measure the surface velocity of natural debris flow is described. This is a simple and interesting technique implemented with a spatial filter constructed as a software program that processes the video image of debris flow instead of a hardware implementation. The surface velocity of the debris flow at the Mt. Yakedake Volcano, Japan, was estimated by this computer-based spatial-filtering method, and the results were compared with those obtained by a hardware-based spatial-filtering method. Computer-based spatial filtering has the important advantage of a capability for tuning the spatial-filter parameters to the target flow. PMID- 18286123 TI - Improved restoration from multiple images of a single object: application to fluorescence microscopy. AB - We present an approach for the combined restoration of multiple different images of a single object. A linear Tikhonov filter adapted for this purpose is derived in detail. Nonlinear constrained algorithms can also be adapted, and we illustrate this possibility for an iterative constrained Tikhonov algorithm. Both the linear and the iterative constrained Tikhonov algorithms were used to analyze performance in fluorescence confocal imaging by use of simulated and experimental data. One can improve the quality of restored confocal images significantly if the signal that normally is rejected by the detection pinhole of a confocal laser scanning microscope is also recorded on a separate detector such that the two recorded signals are used together for image restoration according to the proposed algorithms. PMID- 18286124 TI - Optical Security System with Fourier Plane encoding. AB - We propose a new technique for security verification of personal documents and other forms of personal identifications such as ID cards, passports, or credit cards. In this technique a primary pattern that might be a phase-encoded image is convolved by a random code. The information is phase encoded on the personal document. Therefore the information cannot be reproduced by an intensity detector such as a CCD camera. An optical processor based on the nonlinear joint transform correlator is used to perform the verification and the validation of documents with this technique. By verification of the biometrics information and the random code simultaneously, the proposed optical system determines whether a card is authentic or is being used by an authorized person. We tested the performance of the optical system for security and validation in the presence of input noise and in the presence of distortion of the information on the card. The performance of the proposed method is evaluated by use of a number of metrics. Statistical analysis of the system is performed to investigate the noise tolerance and the discrimination against false inputs for security verification. PMID- 18286125 TI - Rotation-invariant and controllable space-variant optical correlation. AB - We propose a method for designing a correlator for achieving rotation-invariant and controllable space-variant optical correlation. The design concept is based on a combination of fractional correlation and circular-harmonic decomposition of the reference object. The suggested method is described and analyzed in detail. Numerical simulations show that this new correlator might provide potential applications in practice. PMID- 18286126 TI - In-line digital holographic interferometry. AB - An optical system based on in-line digital holography for the evaluation of deformations is described. In-line holograms are recorded on a CCD chip. The problem of overlapping twin images typical for the in-line arrangement is solved by digital reconstruction and filtering of the unwanted wave fronts. Two separate interferograms of an object under test in its undeformed and deformed states are recorded each on a CCD chip. The phases of the two wave fronts are obtained from the complex amplitudes of the digital reconstructed wave fronts, and the deformation is calculated from the phase differences. Experimental results are presented. PMID- 18286127 TI - Reliability Study of Holographic Optical Elements Made with DuPont Photopolymer. AB - We report reliability-test results of transmission-type holographic optical elements (HOE's) made with the DuPont photopolymer HRF-600. The reliability tests performed include 6000 cycles of liquid-to-liquid thermal-shock cycling (-55 degrees C-125 degrees C), 2200 cycles of air-to-air thermal cycling (-55 degrees C-125 degrees C), 1500 h of humidity testing (85 degrees C and a relative humidity of 85%), and 675 h of burn-in testing at 125 degrees C. A total of 210 holograms was tested, with 532 data points collected for diffraction-efficiency measurements. The results show that the average efficiency change after these tests is in the range of -4% to 0% and the standard deviation is only ~10%. PMID- 18286128 TI - Parallel detection algorithm for page-oriented optical memories. AB - We present a parallel algorithm for the reliable detection of two-dimensional binary data in page-oriented memories. The development of the proposed pseudodecision-feedback equalization (PDFE) method is motivated by the classical decision-feedback equalization receiver. The technique takes advantage of the known or the estimated optical system characteristics to mitigate space-variant blur and additive thermal noise. We extend the method to correct for fixed pattern errors including magnification, rotation, and transverse shift. Advantages of the PDFE algorithm include its parallel design, low computational complexity, and local connectivity. A system-capacity metric is used to compare the performance of the PDFE receiver with other conventional approaches, including the simple threshold, the 1:2 modulation code, and the Wiener filter. Results show the PDFE to outperform all the above techniques over a variety of channels for both incoherent and coherent systems. Implementation issues are discussed, and a MOSIS (Metal-Oxide Semiconductor Implementation Service) 2-mum design is presented. PMID- 18286129 TI - Effects of refractive-index mismatch on three-dimensional optical data-storage density in a two-photon bleaching polymer. AB - Reported is an investigation into the effect of spherical aberration caused by the mismatch of the refractive indices between the recording material and its immersion medium on the three-dimensional optical data-storage density in a two photon bleaching polymer. It is found both theoretically and experimentally that spherical aberration can be compensated for by a change in the tube length at which a microscope objective is operated in recording and reading processes. After compensation for the spherical aberration it is possible to achieve a three dimensional recording density of 3.5 Tbits/cm(3) for a commercial objective with a numerical aperture of 1.4. PMID- 18286130 TI - Molecular backscatter heterodyne lidar: a computational evaluation. AB - The application of heterodyne lidar to observe molecular scattering is considered. Despite the reduced Rayleigh cross section, infrared systems are predicted to require mean power levels comparable with those of current and proposed direct detection lidars that operate with the thermally broadened spectra in the visible or ultraviolet. Rayleigh-Brillouin scattering in the kinetic and hydrodynamic (collisional) regimes encountered in the infrared is of particular interest because the observed spectrum approaches a triplet of relatively narrow lines that are more suitable for wind, temperature, and pressure measurements. PMID- 18286131 TI - Hyperspectral remote sensing for shallow waters. I. A semianalytical model. AB - For analytical or semianalytical retrieval of shallow-water bathymetry and/or optical properties of the water column from remote sensing, the contribution to the remotely sensed signal from the water column has to be separated from that of the bottom. The mathematical separation involves three diffuse attenuation coefficients: one for the downwelling irradiance (K(d)), one for the upwelling radiance of the water column (K(u)(C)), and one for the upwelling radiance from bottom reflection (K(u)(B)). Because of the differences in photon origination and path lengths, these three coefficients in general are not equal, although their equality has been assumed in many previous studies. By use of the Hydrolight radiative-transfer numerical model with a particle phase function typical of coastal waters, the remote-sensing reflectance above (R(rs)) and below (r(rs)) the surface is calculated for various combinations of optical properties, bottom albedos, bottom depths, and solar zenith angles. A semianalytical (SA) model for r(rs) of shallow waters is then developed, in which the diffuse attenuation coefficients are explicitly expressed as functions of in-water absorption (a) and backscattering (b(b)). For remote-sensing inversion, parameters connecting R(rs) and r(rs) are also derived. It is found that r(rs) values determined by the SA model agree well with the exact values computed by Hydrolight (~3% error), even for Hydrolight r(rs) values calculated with different particle phase functions. The Hydrolight calculations included b(b)/a values as high as 1.5 to simulate high-turbidity situations that are occasionally found in coastal regions. PMID- 18286132 TI - Correcting global solar ultraviolet spectra recorded by a brewer spectroradiometer for its angular response error. AB - We present a methodology for correcting the global UV spectral measurements of a Brewer MKIII spectroradiometer for the error introduced by the deviation of the angular response of the instrument from the ideal response. This methodology is applicable also to other Brewer spectroradiometers that are currently in operation. The various stages of the methodology are described in detail, together with the uncertainties involved in each stage. Finally global spectral UV measurements with and without the application of the correction are compared with collocated measurements of another spectroradiometer and with model calculations, demonstrating the efficiency of the method. Depending on wavelength and on the aerosol loading, the cosine correction factors range from 2% to 7%. The uncertainties involved in the calculation of these correction factors were found to be relatively small, ranging from ~0.2% to ~2%. PMID- 18286133 TI - Integration of grating couplers in two-story waveguides for rotary displacement sensing. AB - An integrated-optic device, constructed by stacking two types of grating coupler in a two-story structure of waveguides, is proposed for sensing angular displacement of spindle rotation. In the first story a guided wave is diffracted by a grating coupler and becomes a sensing beam. The sensing beam is reflected by a mirror with a quarter-wave plate attached to a spindle head and is coupled back into the second story by another grating coupler. We measured the rotary displacement of the spindle by detecting variation of polarization direction of the reflected beam. A prototype device has been designed and fabricated, and the operation principle is experimentally confirmed. PMID- 18286134 TI - Wavelength selectivity of in-line fiber-optic filter devices. AB - We present an analysis of the performance characteristics of in-line fiber-optic bandpass and band-stop wavelength filters. A planar waveguide model was used for numerical simulations of the device performance. It was recently reported that the band-stop and the bandpass characteristics of a filter device depend on the depth of polishing of a single-mode fiber. However, our numerical simulations show that both types of filter characteristic can be obtained with the same depth of polishing. PMID- 18286135 TI - Fault-tolerant dense multiwavelength add-drop filter with a two-dimensional digital micromirror device. AB - A two-dimensional digital micromirror device- (DMD-) based add-drop filter is introduced for dense wavelength-division multiplexed applications. Features of the filter include polarization-insensitive operation, low interchannel cross talk parallel processing, and a fault-tolerant design. Experiments include evaluation of a parallel-beam-fed interchannel cross-talk with a 25-beam mask and a three-color (red, green, blue) filter operational test that indicates output port average switching optical signal-to-noise and multiwavelength cross-talk ratios of 24 dB. The DMD-based average optical loss for the filter is -2.85 dB for visible-light free-space operation, whereas a -8.88-dB optical loss is measured for a fiber-coupled filter operating at 1319 nm. PMID- 18286136 TI - Nondestructive method for measuring the degree of cluster-induced quenching in er(3+)-doped waveguides and fibers. AB - Ion clustering is one of the major dissipative processes in the operation of Er(3+)-doped planar waveguide amplifiers and heavily doped fiber amplifiers. We propose and demonstrate a nondestructive method for measuring the fraction of erbium ions in doped planar waveguides and fibers that are quenched by ultrafast parasitic energy transfer in ion pairs or clusters. The method is based on unsaturable absorption measurements in the 980-nm absorption band. By combining transmission data at resonance with the corresponding data for radiation outside the absorption band, the influence of background and coupling losses can be eliminated. PMID- 18286137 TI - Performance limitations of a free-space optical communication satellite network owing to vibrations: heterodyne detection. AB - Free-space optical communication between satellites in a distributed network can permit high data rates of communication between different places on Earth. To establish optical communication between any two satellites requires that the line of sight of their optics be aligned during the entire communication time. Because of the large distance between the satellites and the alignment accuracy required, the pointing from one satellite to another is complicated because of vibrations of the pointing system caused by two fundamental stochastic mechanisms: tracking noise created by the electro-optic tracker and vibrations derived from mechanical components. Vibration of the transmitter beam in the receiver plane causes a decrease in the received optical power. Vibrations of the receiver telescope relative to the received beam decrease the heterodyne mixing efficiency. These two factors increase the bit-error rate of a coherent detection network. We derive simple mathematical models of the network bit-error rate versus the system parameters and the transmitter and receiver vibration statistics. An example of a practical optical heterodyne free-space satellite optical communication network is presented. From this research it is clear that even low-amplitude vibration of the satellite-pointing systems dramatically decreases network performance. PMID- 18286138 TI - Reflective integrated optical elements produced by ion exchange in glass. AB - The use of total reflection at the boundary between the guiding region and the substrate (or another guiding region) to implement waveguide achromatic elements is discussed. Two geometries of the boundary, parabolic and elliptical, are proposed and analyzed. The parabolic boundary was fabricated by selective ion exchange, and both its focusing capabilities and the compatibility of its numerical aperture with optical fibers are demonstrated. PMID- 18286139 TI - Hot-press molded poly(methyl methacrylate) matrix for solid-state dye lasers. AB - A hot-press molding method was used to fabricate dye-doped poly(methyl methacrylate) (PMMA) slabs. Three rhodamine dyes, Rh640 (ClO(4)), Rh6G(ClO(4)), and Rh6G (Cl), were impregnated into the PMMA matrix first by dissolving the dye and granular PMMA in a solvent mixture of chloroform and methanol and then heating the mixture in vacuo at 175 degrees C to obtain a spongy preform. The powdered preform was molded into slabs at 175 degrees C and at <1 mbar, to eliminate the formation of bubbles in the slabs. We annealed the slabs for several hours to improve its optical homogeneity and hence its lasing efficiency. When pumped by a 1.5-mJ nitrogen laser, we obtained peak lasing efficiencies of 8% and 7.8%, respectively, for Rh6G (ClO(4)) and Rh640 (ClO(4)) in PMMA matrices. The lasing efficiency of Rh6G (ClO(4))-doped PMMA suffered a reduction rate of 0.012%/shot compared with 0.15%/shot for Rh640 (ClO(4))-doped PMMA. In contrast, Rh6G (Cl) in a hot-press molded PMMA slab suffered thermal bleaching that resulted in a low lasing efficiency of <1%; this can be explained by its absorption and fluorescence characteristics. PMID- 18286140 TI - Axial force acting on a dielectric sphere in a focused laser beam. AB - We present the detailed behavior of the axial force acting on a dielectric sphere exerted by the optical pressure of a focused Gaussian laser beam. Comparison is made between the numerical results and those calculated from the radiation pressure cross section. There is also a discussion as to whether the expressions for the axial force given in this paper are consistent with the previously reported experimental results. Moreover, a simple experimental method to measure the axial force on a polystyrene sphere suspended in water is demonstrated, and fairly good agreement between theoretical and experimental results was obtained. PMID- 18286141 TI - Double-Pass-Pumped Tm:YAG Laser with a Simple Cavity Configuration. AB - For a double-pass-pumped cw Tm:YAG laser, we developed a theoretical model, taking into account reabsorption loss and mode matching between the pumping light and the cavity mode. We also demonstrated that efficient operation can be obtained with a simple cavity configuration by using a dichroic output mirror, highly reflective at the pumping wavelength and partially reflective at the lasing wavelength. Experimental comparisons of this pumping method with single pass pumping showed that the longitudinally double-pass-pumped Tm:YAG laser performs as well at room temperature as the single-pass-pumped laser performs at 10 degrees C. PMID- 18286142 TI - Highly Efficient 68-W Green-Beam Generation by Use of an Intracavity Frequency Doubled Diode Side-Pumped Q-Switched Nd:YAG Rod Laser. AB - An intracavity frequency-doubled diode side-pumped Nd:YAG laser was developed by use of an advanced cavity configuration and a diffusive close-coupled side pumping design. A maximum green power of 68 W was generated at a 20-kHz repetition rate with 18.4% optical-to-optical conversion efficiency and 7.1% electrical-to-optical conversion efficiency. PMID- 18286143 TI - Design of a Triangular Active Ring Laser 13 m on a Side. AB - The design and testing of a proof-of-principle triangular active ring laser interferometer ~13 m on a side is discussed. Issues such as lock in, multimode interference, mode hopping, and neon isotope mixtures are examined as they relate to large He-Ne ring lasers. Responses of the ring laser to Earth's rotation and perturbations that change its tilt or area are presented. Some potential applications are suggested for large interferometers. PMID- 18286144 TI - Frequency Stability at the Kilohertz Level of a Rubidium-Locked Diode Laser at 192.114 THz. AB - The frequency stability of a 1560-nm diode laser, whose second harmonic was locked to (87)Rb sub-Doppler lines, was characterized by measuring the beat frequency relative to a 780-nm reference laser that was locked to sub-Doppler lines of another rubidium cell. The square root of the Allan variance reached a minimum value of 7.5 x 10(-12) in 1 s, which corresponded to frequency variations of 1.44 kHz for the 1560-nm laser. The frequency reproducibility of the system was approximately 1 x 10(-9). These values are better than those that can be achieved by locking to Doppler-broadened transitions at the 1550-nm wavelength band. PMID- 18286145 TI - Working beyond the static limits of laser stability by use of adaptive and polarization-conjugation optics. AB - Strong thermo-optical aberrations in flash-lamp-pumped Nd:Cr:GSGG rods were corrected to yield TEM(00) output at twice the efficiency of Nd:YAG. A hemispherical resonator operating at the limit of stability was employed. As much as 3 W of average power in a Gaussian beam (M(2) approximately 1) was generated. Unique features were zero warm-up time and the ability to vary the repetition rate without varying energy, near- and far-field profiles, or polarization purity. Thermal focusing and astigmatism were corrected with a microprocessor controlled adaptive-optics backmirror composed of discrete elements (variable radius mirror). A reentrant resonator coupled polarizer losses back into the laser rod and corrected depolarization. PMID- 18286146 TI - Simulated dynamic behavior of single and multiple spheres in the trap region of focused laser beams. AB - An enhanced photon propagation method is used to calculate the forces and torque present on each sphere of a system of particles located in the vicinity of focused laser-trapping beams. Infinitesimal trajectory displacements are computed through classical mechanics and the new particle position used to define the next trapping system geometry considered. Repeated applications of the process, implemented as a computer program, enables full trajectory plotting and the dynamic behavior of the systems to be explored as a function of time. PMID- 18286147 TI - Output Energy Fluctuations of the Nd:YAG Amplifier Chain of a Photoinjector Drive Laser System. AB - In the Etude d'un LaSer Accordable, electron bunches consist of trains of picosecond pulses extracted from a photocathode by a drive laser system: This system consists of a mode-locked Nd:YAG oscillator followed by a pulse compressor, an amplifier chain, and a second-harmonic-generation stage. The performance of the linac critically depends on the energy stability of this system. It is demonstrated theoretically and experimentally that the energy fluctuations of the Nd:YAG oscillator are significantly reduced by the amplifier chain, that the remaining energy fluctuations of the whole system are mainly due to amplifier pump fluctuations, and that the amplifier chain can be optimized to reduce the global energy fluctuations from 1.5% rms to <1% rms. PMID- 18286148 TI - Diode-Pumped Tunable Room-Temperature LiF:F(2)(-) Color-Center Laser. AB - Pulsed room-temperature tunable operation of a diode-pumped LiF:F(2)(-) laser has been demonstrated for the first time to the author's knowledge. The LiF:F(2)(-) oscillator was tunable in the 1122-1201-nm range. In the free-running mode, with a nonselective resonator, the LiF:F(2)(-) laser showed a 4% optical-to-optical conversion efficiency and a measured 2.5% pumping-to-lasing efficiency at the maximum of the tuning curve for tunable narrow-band operation. PMID- 18286149 TI - All-Solid-State Tunable Ultraviolet Subnanosecond Laser with Direct Pumping by the Fifth Harmonic of a Nd:YAG laser. AB - We report what we believe is the first all-solid-state tunable ultraviolet laser pumped by the fifth harmonic of a Q-switched Nd:YAG laser. Our laser based on a Ce(3+):LiLuF(4) active medium stably generates a single, satellite-free, 0.88-ns pulse under 5-ns, 10-Hz repetition rate pumping conditions. A novel tilted incident-angle side-pumping scheme resulted in a simple laser-cavity design. PMID- 18286150 TI - Dye photodestruction in a solid-state dye laser with a polymeric gain medium. AB - The process of dye photodestruction in a solid-state dye laser is studied, and implemented is a polymeric gain medium doped with a strongly concentrated dye. The behavior of the conversion efficiency in the polymeric gain medium pumped with different laser-pulse repetition rates and the process of dye photobleaching are analyzed. The contribution of the heating of the host material into the dye molecules' deactivation is discussed. The negative effect of high dye concentration on the dye stability under a high pump repetition rate is reported and analyzed for the first time to my knowledge. A comparison of the present results with recently published data demonstrates the major role of photodestruction, rather than direct thermodestruction, in the dye stability of the solid-state gain medium. The role of additives with low molecular weights in the polymeric matrix, for increasing the stability of the gain material, is discussed. PMID- 18286151 TI - Coefficients of a Dispersion Equation for the Birefringence of a CdGeAs(2) Nonlinear Crystal at Different Temperatures. AB - Birefringence (B(f)) and its temperature derivative (dB(f)/dT) determine the temperature characteristics of nonlinear optical laser devices. The birefringence and its dispersion for a CdGeAs(2) nonlinear crystal are analyzed critically by use of a recently introduced, physically meaningful, dispersion equation at different operating temperatures from 14 to 450 K, which fits reasonably well with the experimental data. This equation is based on the average electronic absorption gap and the lattice absorption gap at the short- and long-wavelength transparency limits, respectively. PMID- 18286152 TI - Broadband Tuning of a Continuous-Wave, Doubly Resonant, Lithium Triborate Optical Parametric Oscillator from 791 to 1620 nm. AB - We report the use of extreme broadband, high reflectivity >99.5%, optical parametric oscillator (OPO) cavity mirrors. A continuous-wave, doubly resonant, OPO demonstrated tuning over a range of 791-1620 nm with a single mirror set. Wavelength tuning was performed by temperature tuning the nonlinear material of lithium triborate. Narrow linewidth oscillation was confirmed throughout the tuning range, and threshold pump power increased gradually from 50 mW near the degeneracy to 800 mW at the tuning band edge in a double-pass pumping configuration. PMID- 18286153 TI - Spectral Measurement of the Film-Substrate Index Difference in Proton-Exchanged LiNbO(3) Waveguides. AB - We report the spectral characterization of proton-exchanged lithium niobate (PE:LiNbO(3)) waveguides in terms of the variation of the refractive-index difference between the waveguiding layer and the substrate. The dispersion of the extraordinary refractive-index increase (deltan(e)) is measured from 405 to 1319 nm with several light sources. Two types of proton-exchanged waveguide, prepared under different conditions, are studied. These measurements should be of use in the optimization of PE:LiNbO(3) waveguides for nonlinear optical applications, particularly in second-harmonic generation in the blue-green wavelength region. PMID- 18286154 TI - Lidar-signal compression by photomultiplier gain modulation: influence of detector nonlinearity. AB - The application of photomultiplier gain modulation to the compression of wide dynamic-range lidar signals is investigated in relation to the effect of the gain level on anode-signal linearity. Gain reduction is achieved by the coupling of modulation signals through either multidynode or focus-grid gating networks. This technique facilitates signal recovery and prevents detector nonlinearity and dynode damage caused by high near-field lidar signals. The measurements were performed in the current mode primarily on a 50-mm-diameter, 12-stage photomultiplier (EMI 9214) with a bialkali photocathode. With 3- or 4-dynode based modulation made at a photomultiplier voltage of 1300 V and a gain of 1 x 10(7), signals of ~6 mA can be maintained at the 1% linearity limit from 100% to 0.2% modulation, corresponding to a 500-fold reduction in the lidar-signal dynamic range. A significant advantage to dynode modulation is that it preserves the shot-signal-to-noise ratio of the incoming signal, which is not true for focus-grid modulation or external predetection schemes such as controlled obscuration or Pockels-cell modulation that attenuate the as-yet unamplified signal. PMID- 18286155 TI - Modeling of direct detection Doppler wind lidar. I. The edge technique. AB - Analytic models, based on a convolution of a Fabry-Perot etalon transfer function with a Gaussian spectral source, are developed for the shot-noise-limited measurement precision of Doppler wind lidars based on the edge filter technique by use of either molecular or aerosol atmospheric backscatter. The Rayleigh backscatter formulation yields a map of theoretical sensitivity versus etalon parameters, permitting design optimization and showing that the optimal system will have a Doppler measurement uncertainty no better than approximately 2.4 times that of a perfect, lossless receiver. An extension of the models to include the effect of limited etalon aperture leads to a condition for the minimum aperture required to match light collection optics. It is shown that, depending on the choice of operating point, the etalon aperture finesse must be 4-15 to avoid degradation of measurement precision. A convenient, closed-form expression for the measurement precision is obtained for spectrally narrow backscatter and is shown to be useful for backscatter that is spectrally broad as well. The models are extended to include extrinsic noise, such as solar background or the Rayleigh background on an aerosol Doppler lidar. A comparison of the model predictions with experiment has not yet been possible, but a comparison with detailed instrument modeling by McGill and Spinhirne shows satisfactory agreement. The models derived here will be more conveniently implemented than McGill and Spinhirne's and more readily permit physical insights to the optimization and limitations of the double-edge technique. PMID- 18286157 TI - Electromagnetic scattering by an aggregate of spheres: errata. PMID- 18286156 TI - Modeling of Direct Detection Doppler Wind Lidar. II. The Fringe Imaging Technique. AB - A simple analytic model is developed for the shot-noise-limited measurement precision of Doppler wind lidars based on the fringe imaging technique by use of either molecular or aerosol atmospheric backscatter. The model leads to etalon design parameters for an instrument optimized for precision. The ultimate measurement precision possible is two to four times the limit for a perfect, lossless receiver. The corresponding result for the double-edge Doppler analyzer was a ratio of 2.5, showing that the two methods are little different in this respect. For aerosol backscatter instruments, the wind speed dynamic range of the fringe imager is substantially greater than that for the edge detector. The etalon aperture needed to meet system etendue requirements is derived and shown to be approximately half that of each of the two etalons required by the double edge technique. A comparison with more detailed modeling of fringe imaging Doppler-shift analyzers shows good agreement for the Rayleigh model and fair for the aerosol version, confirming the validity of this simpler technique for analyzer design and performance prediction. PMID- 18286158 TI - Experimental investigation of the influence of the relative position of the scattering layer on image quality: the shower curtain effect. AB - The imaging quality of optical systems in a turbid environment is influenced not only by the content of the turbid layer between the object and the optical receiver but also by the inhomogeneity of that medium. This is important, particularly when imaging is performed through clouds, nonhomogeneous layers of dust, or over vertical or slant paths through the atmosphere. Forward small-angle scattering influences image quality and blur more severely when the scattering layer is closer to the receiver. In this study it is the influence of the relative position of the scattering layer on the image quality and modulation transfer function (MTF) that is investigated. The scattering layer in controlled laboratory experiments consists of calibrated polystyrene particles of known size and quantity in a small cuvette. A point source was imaged by a computerized imaging system through a layer containing polystyrene particles, and the point spread function (PSF) was recorded. The aerosol MTF was calculated using the measured PSF. The MTF was measured as a function of changing relative distance of the scattering layer from the receiver, whereas the object-plane-to-receiver distance was constant. The experimental results were compared to theoretical shower curtain effect models based on the solution from radiative transfer theory under the small-angle approximation. Although the general trend of the experimental results certainly agrees with the theoretical models, it could be that the small-angle approximation method might be of limited validity at such low spatial frequencies. Aggregation also causes some disagreement with predictions from theory. PMID- 18286159 TI - Multiple-scattering effect on ozone retrieval from space-based differential absorption lidar measurements. AB - Single-scattering and multiple-scattering lidar signals are calculated for a spaceborne differential absorption lidar system for global ozone measurements at the on and off wavelength pair at 305 and 315 nm. The effect of multiple scattering is found to be negligible on stratospheric and tropospheric ozone retrieval under background stratospheric aerosol. Under low-visibility conditions in the planetary boundary layer the presence of multiple scattering causes an overestimation in maritime aerosol and an underestimation in urban as well as in rural aerosol. This effect is also examined in three cirrus models. The multiple scattering does not permit accurate ozone retrieval within cirrus; however, below it the solution recovers somewhat with generally an underestimation depending on the type and density of cirrus. The effect of aerosol and Rayleigh extinction on the ozone retrieval is also discussed. PMID- 18286160 TI - Effective suppression of multiply scattered light in static and dynamic light scattering. AB - The evaluation of conventional light-scattering experiments in turbid media is often highly complicated because of the presence of multiple scattering contributions. The three-dimensional (3-D) cross-correlation method presented provides an effective and handy method to suppress the influence of multiply scattered light. As the time dependence of the 3-D cross-correlation function is determined solely by the singly scattered light, the evaluation of the decay constant yields reliable values for the effective diffusion coefficient and the hydrodynamic particle size of the suspended particles. Furthermore, analysis of the amplitude of the 3-D cross-correlation function permits the determination of the differential scattering cross section even for highly turbid suspensions. PMID- 18286161 TI - Influence of bubbles on scattering of light in the ocean. AB - The scattering and backscattering properties of bubble populations in the upper ocean are estimated with Mie theory and a generalized bubble size spectrum based on in situ observations. Optical properties of both clean bubbles and bubbles coated with an organic film are analyzed; the results are compared with the corresponding optical properties of micro-organisms of similar size. Given a bubble number density (from ~10(5) to ~10(7) m(-3)) frequently found at sea, the bubble populations significantly influence the scattering process in the ocean, especially in oligotrophic waters. Bubbles appear to make a large contribution to the missing terms in constructing the observed total backscattering coefficient of the ocean. This contribution to backscattering is strongly enhanced if the bubbles are coated with organic film. The injection of bubbles will shift ocean color toward the green, resembling phytoplankton blooms, and hence introducing error in ocean color remote sensing if its effect is not corrected. PMID- 18286162 TI - Effect of apodization on the retrieval of geophysical parameters from fourier transform spectrometers. AB - The problem of the effect of apodization on the retrieval of geophysical parameters from infrared radiances recorded by Fourier transform spectrometers has been analytically and numerically addressed. Exploiting a matrix representation of apodization, we first derive a general analytical expression for the apodized covariance matrix and then show that apodization, when properly applied, has no effect on retrievals. The methodology has been applied to investigate the effect of Gaussian apodization on the Infrared Atmospheric Sounding Interferometer currently under development at the laboratories of the French Space Agency. PMID- 18286163 TI - Detector Nonlinearity Correction Scheme for the LPMA Balloonborne Fourier Transform Spectrometer. AB - The Limb Profile Monitor of the Atmosphere (LPMA) instrument is a Fourier transform spectrometer designed to record stratospheric (and in some cases tropospheric) absorption spectra from a balloon gondola. This spectrometer operates with two-detector output optics (photoconductive HgCdTe and photovoltaic InSb, liquid-nitrogen cooled). The response of the HgCdTe detector becomes nonlinear for high photon fluxes, which is the case for solar occultation. We have designed a processing scheme, based on the minimization of out-of-optical band spectral artifacts, to correct for the effect of nonlinearity in the useful spectral range. The method is explained, and sample results are presented for spectra recorded in different balloon flight conditions and with two different HgCdTe detectors. PMID- 18286164 TI - Differential absorption mueller matrix spectroscopy and the infrared detection of crystalline organics. AB - The complete 16-element Mueller matrices for backscattering from amino acids, sugars, and other enantiomorphic compounds pressed into wafer form were measured at infrared wavelengths. For each compound a pair of CO(2) laser lines was selected from the 9.1-11.6-mum region such that one line excited an absorption band in the compound, whereas the other did not. It was observed that at least some of the matrix elements differed significantly depending on which of the two wavelengths was used in the measurement. We propose that a neural network pattern recognition system can be trained to detect the presence of specific compounds based on multiwavelength backscatter Mueller matrix measurements. PMID- 18286165 TI - Isotope (18)O/(16)O Ratio Measurements of Water Vapor by use of Photoacoustic Spectroscopy. AB - We applied a photoacoustic spectroscopy technique to isotope ratio measurements of (16)O and (18)O in water-vapor samples, using a pulsed tunable dye laser pumped by a Nd:YAG laser. The fourth overtone bands (4nu(OH)) of water molecules near 720 nm were investigated. We identified the absorption lines of H(2)(16)O and H(2)(18)O in the photoacoustic spectra that we measured by using an (18)O enriched water sample and the HITRAN database. We measured the difference in the (18)O/(16)O isotope ratios for normal distilled water and Antarctic ice, using the photoacoustic method. The value obtained for the difference between the two samples is delta(18)O = -32 ? 16 per thousand, where the indicated deviation was a 1varsigma value among 240-s measurements, whereas the value measured with a conventional isotope mass spectrometer was delta(18)O = -28 ? 2 per thousand. This method is demonstrated to have the potential of a transportable system for in situ and quick measurements of the H(2)(18)O/H(2)(16)O ratio in the environment. PMID- 18286166 TI - Molecular pathways involved in loss of kidney graft function with tubular atrophy and interstitial fibrosis. AB - Loss of kidney graft function with tubular atrophy (TA) and interstitial fibrosis (IF) causes most kidney allograft losses. We aimed to identify the molecular pathways involved in IF/TA progression. Kidney biopsies from normal kidneys (n = 24), normal allografts (n = 6), and allografts with IF/TA (n = 17) were analyzed using high-density oligonucleotide microarray. Probe set level tests of hypotheses tests were conducted to identify genes with a significant trend in gene expression across the three groups using Jonckheere-Terpstra test for trend. Interaction networks and functional analysis were used. An unsupervised hierarchical clustering analysis showed that all the IF/TA samples were associated with high correlation. Gene ontology classified the differentially expressed genes as related to immune response, inflammation, and matrix deposition. Chemokines (CX), CX receptor (for example, CCL5 and CXCR4), interleukin, and interleukin receptor (for example, IL-8 and IL10RA) genes were overexpressed in IF/TA samples compared with normal allografts and normal kidneys. Genes involved in apoptosis (for example, CASP4 and CASP5) were importantly overexpressed in IF/TA. Genes related to angiogenesis (for example, ANGPTL3, ANGPT2, and VEGF) were downregulated in IF/TA. Genes related to matrix production-deposition were upregulated in IF/TA. A distinctive gene expression pattern was observed in IF/TA samples compared with normal allografts and normal kidneys. We were able to establish a trend in gene expression for genes involved in different pathways among the studied groups. The top-scored networks were related to immune response, inflammation, and cell-to-cell interaction, showing the importance of chronic inflammation in progressive graft deterioration. PMID- 18286167 TI - Transparent meta-analysis of prospective memory and aging. AB - Prospective memory (ProM) refers to our ability to become aware of a previously formed plan at the right time and place. After two decades of research on prospective memory and aging, narrative reviews and summaries have arrived at widely different conclusions. One view is that prospective memory shows large age declines, larger than age declines on retrospective memory (RetM). Another view is that prospective memory is an exception to age declines and remains invariant across the adult lifespan. The present meta-analysis of over twenty years of research settles this controversy. It shows that prospective memory declines with aging and that the magnitude of age decline varies by prospective memory subdomain (vigilance, prospective memory proper, habitual prospective memory) as well as test setting (laboratory, natural). Moreover, this meta-analysis demonstrates that previous claims of no age declines in prospective memory are artifacts of methodological and conceptual issues afflicting prior research including widespread ceiling effects, low statistical power, age confounds, and failure to distinguish between various subdomains of prospective memory (e.g., vigilance and prospective memory proper). PMID- 18286168 TI - Poor reporting of scientific leadership information in clinical trial registers. AB - BACKGROUND: In September 2004, the International Committee of Medical Journal Editors (ICMJE) issued a Statement requiring that all clinical trials be registered at inception in a public register in order to be considered for publication. The World Health Organization (WHO) and ICMJE have identified 20 items that should be provided before a trial is considered registered, including contact information. Identifying those scientifically responsible for trial conduct increases accountability. The objective is to examine the proportion of registered clinical trials providing valid scientific leadership information. METHODOLOGY/PRINCIPAL FINDINGS: We reviewed clinical trial entries listing Canadian investigators in the two largest international and public trial registers, the International Standard Randomized Controlled Trial Number (ISRCTN) register, and ClinicalTrials.gov. The main outcome measures were the proportion of clinical trials reporting valid contact information for the trials' Principal Investigator (PI)/Co-ordinating Investigator/Study Chair/Site PI, and trial e mail contact address, stratified by funding source, recruiting status, and register. A total of 1388 entries (142 from ISRCTN and 1246 from ClinicalTrials.gov) comprised our sample. We found non-compliance with mandatory registration requirements regarding scientific leadership and trial contact information. Non-industry and partial industry funded trials were significantly more likely to identify the individual responsible for scientific leadership (OR = 259, 95% CI: 95-701) and to provide a contact e-mail address (OR = 9.6, 95% CI: 6.6-14) than were solely industry funded trials. CONCLUSIONS/SIGNIFICANCE: Despite the requirements set by WHO and ICMJE, data on scientific leadership and contact e-mail addresses are frequently omitted from clinical trials registered in the two leading public clinical trial registers. To promote accountability and transparency in clinical trials research, public clinical trials registers should ensure adequate monitoring of trial registration to ensure completion of mandatory contact information fields identifying scientific leadership. PMID- 18286169 TI - FOXP3 promoter demethylation reveals the committed Treg population in humans. AB - BACKGROUND: Naturally occurring thymus derived regulatory T cells (Tregs) are central in the maintenance of self-tolerance. The transcription factor FOXP3 is crucial for the suppressive activity of Tregs and is considered the most specific marker for this population. However, human non regulatory T cells upregulate FOXP3 transiently upon activation which calls for other means to identify the Treg population. Since epigenetic mechanisms are involved in the establishment of stable gene expression patterns during cell differentiation, we hypothesized that the methylation profile of the FOXP3 promoter would allow the distinction of truly committed Tregs. METHODOLOGY/PRINCIPAL FINDINGS: Human CD4(+)CD25(hi) Tregs displayed a demethylated FOXP3 promoter (1.4%+/-0.95% SEM methylated) in contrast to CD4(+)CD25(lo) T cells which were partially methylated (27.9%+/-7.1%). Furthermore, stimulated CD4(+)CD25(lo) T cells transiently expressed FOXP3 but remained partially methylated, suggesting promoter methylation as a mechanism for regulation of stable FOXP3 expression and Treg commitment. In addition, transient FOXP3 expressing cells exhibited suppressive abilities that correlate to the methylation status of the FOXP3 promoter. As an alternative to bisulphite sequencing, we present a restriction enzyme based screening method for the identification of committed Tregs and apply this method to evaluate the effect of various culturing conditions. We show that a partial demethylation occurs in long term cultures after activation, whereas the addition of TGF-beta and/or IL-10 does not induce any additional change in methylation level. CONCLUSIONS/SIGNIFICANCE: The unique FOXP3 promoter methylation profile in Tregs suggests that a demethylated pattern is a prerequisite for stable FOXP3 expression and suppressive phenotype. Presently, FOXP3 is used to identify Tregs in several human diseases and there are future implications for adoptive Treg transfer in immunotherapy. In these settings there is a need to distinguish true Tregs from transiently FOXP3(+) activated T cells. The screening method we present allows this distinction and enables the identification of cells suitable for in vitro expansions and clinical use. PMID- 18286170 TI - Drosophila muscleblind is involved in troponin T alternative splicing and apoptosis. AB - BACKGROUND: Muscleblind-like proteins (MBNL) have been involved in a developmental switch in the use of defined cassette exons. Such transition fails in the CTG repeat expansion disease myotonic dystrophy due, in part, to sequestration of MBNL proteins by CUG repeat RNA. Four protein isoforms (MblA-D) are coded by the unique Drosophila muscleblind gene. METHODOLOGY/PRINCIPAL FINDINGS: We used evolutionary, genetic and cell culture approaches to study muscleblind (mbl) function in flies. The evolutionary study showed that the MblC protein isoform was readily conserved from nematods to Drosophila, which suggests that it performs the most ancestral muscleblind functions. Overexpression of MblC in the fly eye precursors led to an externally rough eye morphology. This phenotype was used in a genetic screen to identify five dominant suppressors and 13 dominant enhancers including Drosophila CUG-BP1 homolog aret, exon junction complex components tsunagi and Aly, and pro-apoptotic genes Traf1 and reaper. We further investigated Muscleblind implication in apoptosis and splicing regulation. We found missplicing of troponin T in muscleblind mutant pupae and confirmed Muscleblind ability to regulate mouse fast skeletal muscle Troponin T (TnnT3) minigene splicing in human HEK cells. MblC overexpression in the wing imaginal disc activated apoptosis in a spatially restricted manner. Bioinformatics analysis identified a conserved FKRP motif, weakly resembling a sumoylation target site, in the MblC-specific sequence. Site-directed mutagenesis of the motif revealed no change in activity of mutant MblC on TnnT3 minigene splicing or aberrant binding to CUG repeat RNA, but altered the ability of the protein to form perinuclear aggregates and enhanced cell death-inducing activity of MblC overexpression. CONCLUSIONS/SIGNIFICANCE: Taken together our genetic approach identify cellular processes influenced by Muscleblind function, whereas in vivo and cell culture experiments define Drosophila troponin T as a new Muscleblind target, reveal a potential involvement of MblC in programmed cell death and recognize the FKRP motif as a putative regulator of MblC function and/or subcellular location in the cell. PMID- 18286171 TI - A non-human primate model for gluten sensitivity. AB - BACKGROUND AND AIMS: Gluten sensitivity is widespread among humans. For example, in celiac disease patients, an inflammatory response to dietary gluten leads to enteropathy, malabsorption, circulating antibodies against gluten and transglutaminase 2, and clinical symptoms such as diarrhea. There is a growing need in fundamental and translational research for animal models that exhibit aspects of human gluten sensitivity. METHODS: Using ELISA-based antibody assays, we screened a population of captive rhesus macaques with chronic diarrhea of non infectious origin to estimate the incidence of gluten sensitivity. A selected animal with elevated anti-gliadin antibodies and a matched control were extensively studied through alternating periods of gluten-free diet and gluten challenge. Blinded clinical and histological evaluations were conducted to seek evidence for gluten sensitivity. RESULTS: When fed with a gluten-containing diet, gluten-sensitive macaques showed signs and symptoms of celiac disease including chronic diarrhea, malabsorptive steatorrhea, intestinal lesions and anti-gliadin antibodies. A gluten-free diet reversed these clinical, histological and serological features, while reintroduction of dietary gluten caused rapid relapse. CONCLUSIONS: Gluten-sensitive rhesus macaques may be an attractive resource for investigating both the pathogenesis and the treatment of celiac disease. PMID- 18286172 TI - The effect of inappropriate calibration: three case studies in molecular ecology. AB - Time-scales estimated from sequence data play an important role in molecular ecology. They can be used to draw correlations between evolutionary and palaeoclimatic events, to measure the tempo of speciation, and to study the demographic history of an endangered species. In all of these studies, it is paramount to have accurate estimates of time-scales and substitution rates. Molecular ecological studies typically focus on intraspecific data that have evolved on genealogical scales, but often these studies inappropriately employ deep fossil calibrations or canonical substitution rates (e.g., 1% per million years for birds and mammals) for calibrating estimates of divergence times. These approaches can yield misleading estimates of molecular time-scales, with significant impacts on subsequent evolutionary and ecological inferences. We illustrate this calibration problem using three case studies: avian speciation in the late Pleistocene, the demographic history of bowhead whales, and the Pleistocene biogeography of brown bears. For each data set, we compare the date estimates that are obtained using internal and external calibration points. In all three cases, the conclusions are significantly altered by the application of revised, internally-calibrated substitution rates. Collectively, the results emphasise the importance of judicious selection of calibrations for analyses of recent evolutionary events. PMID- 18286173 TI - MAO-B elevation in mouse brain astrocytes results in Parkinson's pathology. AB - Age-related increases in monoamine oxidase B (MAO-B) may contribute to neurodegeneration associated with Parkinson's disease (PD). The MAO-B inhibitor deprenyl, a long-standing antiparkinsonian therapy, is currently used clinically in concert with the dopamine precursor L-DOPA. Clinical studies suggesting that deprenyl treatment alone is not protective against PD associated mortality were targeted to symptomatic patients. However, dopamine loss is at least 60% by the time PD is symptomatically detectable, therefore lack of effect of MAO-B inhibition in these patients does not negate a role for MAO-B in pre-symptomatic dopaminergic loss. In order to directly evaluate the role of age-related elevations in astroglial MAO-B in the early initiation or progression of PD, we created genetically engineered transgenic mice in which MAO-B levels could be specifically induced within astroglia in adult animals. Elevated astrocytic MAO-B mimicking age related increase resulted in specific, selective and progressive loss of dopaminergic neurons in the substantia nigra (SN), the same subset of neurons primarily impacted in the human condition. This was accompanied by other PD-related alterations including selective decreases in mitochondrial complex I activity and increased mitochondrial oxidative stress. Along with a global astrogliosis, we observed local microglial activation within the SN. These pathologies correlated with decreased locomotor activity. Importantly, these events occurred even in the absence of the PD-inducing neurotoxin MPTP. Our data demonstrates that elevation of murine astrocytic MAO-B by itself can induce several phenotypes of PD, signifying that MAO-B could be directly involved in multiple aspects of disease neuropathology. Mechanistically this may involve increases in membrane permeant H(2)O(2) which can oxidize dopamine within dopaminergic neurons to dopaminochrome which, via interaction with mitochondrial complex I, can result in increased mitochondrial superoxide. Our inducible astrocytic MAO-B transgenic provides a novel model for exploring pathways involved in initiation and progression of several key features associated with PD pathology and for therapeutic drug testing. PMID- 18286174 TI - Ybp2 associates with the central kinetochore of Saccharomyces cerevisiae and mediates proper mitotic progression. AB - The spindle checkpoint ensures the accurate segregation of chromosomes by monitoring the status of kinetochore attachment to microtubules. Simultaneous mutations in one of several kinetochore and cohesion genes and a spindle checkpoint gene cause a synthetic-lethal or synthetic-sick phenotype. A synthetic genetic array (SGA) analysis using a mad2Delta query mutant strain of yeast identified YBP2, a gene whose product shares sequence similarity with the product of YBP1, which is required for H(2)O(2)-induced oxidation of the transcription factor Yap1. ybp2Delta was sensitive to benomyl and accumulated at the mitotic stage of the cell cycle. Ybp2 physically associates with proteins of the COMA complex (Ctf19, Okp1, Mcm21, and Ame1) and 3 components of the Ndc80 complex (Ndc80, Nuf2, and Spc25 but not Spc24) in the central kinetochore and with Cse4 (the centromeric histone and CENP-A homolog). Chromatin-immunoprecipitation analyses revealed that Ybp2 associates specifically with CEN DNA. Furthermore, ybp2Delta showed synthetic-sick interactions with mutants of the genes that encode the COMA complex components. Ybp2 seems to be part of a macromolecular kinetochore complex and appears to contribute to the proper associations among the central kinetochore subcomplexes and the kinetochore-specific nucleosome. PMID- 18286175 TI - Shrimps down under: evolutionary relationships of subterranean crustaceans from Western Australia (Decapoda: Atyidae: Stygiocaris). AB - BACKGROUND: We investigated the large and small scale evolutionary relationships of the endemic Western Australian subterranean shrimp genus Stygiocaris (Atyidae) using nuclear and mitochondrial genes. Stygiocaris is part of the unique cave biota of the coastal, anchialine, limestones of the Cape Range and Barrow Island, most of whose nearest evolutionary relations are found in coastal caves of the distant North Atlantic. The dominance of atyids in tropical waters and their food resources suggest they are pivotal in understanding these groundwater ecosystems. METHODOLOGY/PRINCIPLE FINDINGS: Our nuclear and mitochondrial analyses all recovered the Mexican cave genus Typhlatya as the sister taxon of Stygiocaris, rather than any of the numerous surface and cave atyids from Australia or the Indo-Pacific region. The two described Stygiocaris species were recovered as monophyletic, and a third, cryptic, species was discovered at a single site, which has very different physiochemical properties from the sites hosting the two described species. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that Stygiocaris and Typhlatya may descend from a common ancestor that lived in the coastal marine habitat of the ancient Tethys Sea, and were subsequently separated by plate tectonic movements. This vicariant process is commonly thought to explain the many disjunct anchialine faunas, but has rarely been demonstrated using phylogenetic techniques. The Cape Range's geological dynamism, which is probably responsible for the speciation of the various Stygiocaris species, has also led to geographic population structure within species. In particular, Stygiocaris lancifera is split into northern and southern groups, which correspond to population splits within other sympatric subterranean taxa. PMID- 18286176 TI - The neglected intrinsic resistome of bacterial pathogens. AB - Bacteria with intrinsic resistance to antibiotics are a worrisome health problem. It is widely believed that intrinsic antibiotic resistance of bacterial pathogens is mainly the consequence of cellular impermeability and activity of efflux pumps. However, the analysis of transposon-tagged Pseudomonas aeruginosa mutants presented in this article shows that this phenotype emerges from the action of numerous proteins from all functional categories. Mutations in some genes make P. aeruginosa more susceptible to antibiotics and thereby represent new targets. Mutations in other genes make P. aeruginosa more resistant and therefore define novel mechanisms for mutation-driven acquisition of antibiotic resistance, opening a new research field based in the prediction of resistance before it emerges in clinical environments. Antibiotics are not just weapons against bacterial competitors, but also natural signalling molecules. Our results demonstrate that antibiotic resistance genes are not merely protective shields and offer a more comprehensive view of the role of antibiotic resistance genes in the clinic and in nature. PMID- 18286177 TI - The leishmania ARL-1 and Golgi traffic. AB - We present here the characterisation of the Leishmania small G protein ADP Ribosylation Factor-Like protein 1 (ARL-1). The ARL-1 gene is present in one copy per haploid genome and conserved among trypanosomatids. It encodes a protein of 20 kDa, which is equally expressed in the insect promastigote and mammalian amastigote forms of the parasite. ARL-1 localises to the Trans-Golgi Network (TGN); N-terminal myristoylation is essential for TGN localisation. In vivo expression of the LdARL-1/Q74L and LdARL-1/T51N mutants (GTP- and GDP-bound blocked forms respectively) shows that GDP/GTP cycling occurs entirely within the TGN. This is contrary to previous reports in yeast and mammals, where the mutant empty form devoid of nucleotide has been considered as the GDP-blocked form. The dominant-negative empty form mutant LdARL-1/T34N inhibits endocytosis and intracellular trafficking from the TGN to the Lysosome/Multivesicular Tubule and to the acidocalcisomes; these defects are probably related to a mislocalisation of the GRIP domain-containing vesicle tethering factors which cannot be recruited to the TGN by the cytoplasmic LdARL-1/T34N. Thus, besides the functional characterization of a new mutant and a better understanding of ARL-1 GDP/GTP cycling, this work shows that Leishmania ARL-1 is a key component of an essential pathway worth future study. PMID- 18286178 TI - C-ME: a 3D community-based, real-time collaboration tool for scientific research and training. AB - The need for effective collaboration tools is growing as multidisciplinary proteome-wide projects and distributed research teams become more common. The resulting data is often quite disparate, stored in separate locations, and not contextually related. Collaborative Molecular Modeling Environment (C-ME) is an interactive community-based collaboration system that allows researchers to organize information, visualize data on a two-dimensional (2-D) or three dimensional (3-D) basis, and share and manage that information with collaborators in real time. C-ME stores the information in industry-standard databases that are immediately accessible by appropriate permission within the computer network directory service or anonymously across the internet through the C-ME application or through a web browser. The system addresses two important aspects of collaboration: context and information management. C-ME allows a researcher to use a 3-D atomic structure model or a 2-D image as a contextual basis on which to attach and share annotations to specific atoms or molecules or to specific regions of a 2-D image. These annotations provide additional information about the atomic structure or image data that can then be evaluated, amended or added to by other project members. PMID- 18286179 TI - Sequence evidence in the archaeal genomes that tRNAs emerged through the combination of ancestral genes as 5' and 3' tRNA halves. AB - The discovery of separate 5' and 3' halves of transfer RNA (tRNA) molecules-so called split tRNA-in the archaeal parasite Nanoarchaeum equitans made us wonder whether ancestral tRNA was encoded on 1 or 2 genes. We performed a comprehensive phylogenetic analysis of tRNAs in 45 archaeal species to explore the relationship between the three types of tRNAs (nonintronic, intronic and split). We classified 1953 mature tRNA sequences into 22 clusters. All split tRNAs have shown phylogenetic relationships with other tRNAs possessing the same anticodon. We also mimicked split tRNA by artificially separating the tRNA sequences of 7 primitive archaeal species at the anticodon and analyzed the sequence similarity and diversity of the 5' and 3' tRNA halves. Network analysis revealed specific characteristics of and topological differences between the 5' and 3' tRNA halves: the 5' half sequences were categorized into 6 distinct groups with a sequence similarity of >80%, while the 3' half sequences were categorized into 9 groups with a higher sequence similarity of >88%, suggesting different evolutionary backgrounds of the 2 halves. Furthermore, the combinations of 5' and 3' halves corresponded with the variation of amino acids in the codon table. We found not only universally conserved combinations of 5'-3' tRNA halves in tRNA(iMet), tRNA(Thr), tRNA(Ile), tRNA(Gly), tRNA(Gln), tRNA(Glu), tRNA(Asp), tRNA(Lys), tRNA(Arg) and tRNA(Leu) but also phylum-specific combinations in tRNA(Pro), tRNA(Ala), and tRNA(Trp). Our results support the idea that tRNA emerged through the combination of separate genes and explain the sequence diversity that arose during archaeal tRNA evolution. PMID- 18286180 TI - Asap: a framework for over-representation statistics for transcription factor binding sites. AB - BACKGROUND: In studies of gene regulation the efficient computational detection of over-represented transcription factor binding sites is an increasingly important aspect. Several published methods can be used for testing whether a set of hypothesised co-regulated genes share a common regulatory regime based on the occurrence of the modelled transcription factor binding sites. However there is little or no information available for guiding the end users choice of method. Furthermore it would be necessary to obtain several different software programs from various sources to make a well-founded choice. METHODOLOGY: We introduce a software package, Asap, for fast searching with position weight matrices that include several standard methods for assessing over-representation. We have compared the ability of these methods to detect over-represented transcription factor binding sites in artificial promoter sequences. Controlling all aspects of our input data we are able to identify the optimal statistics across multiple threshold values and for sequence sets containing different distributions of transcription factor binding sites. CONCLUSIONS: We show that our implementation is significantly faster than more naive scanning algorithms when searching with many weight matrices in large sequence sets. When comparing the various statistics, we show that those based on binomial over-representation and Fisher's exact test performs almost equally good and better than the others. An online server is available at http://servers.binf.ku.dk/asap/. PMID- 18286182 TI - Early development of the central and peripheral nervous systems is coordinated by Wnt and BMP signals. AB - The formation of functional neural circuits that process sensory information requires coordinated development of the central and peripheral nervous systems derived from neural plate and neural plate border cells, respectively. Neural plate, neural crest and rostral placodal cells are all specified at the late gastrula stage. How the early development of the central and peripheral nervous systems are coordinated remains, however, poorly understood. Previous results have provided evidence that at the late gastrula stage, graded Wnt signals impose rostrocaudal character on neural plate cells, and Bone Morphogenetic Protein (BMP) signals specify olfactory and lens placodal cells at rostral forebrain levels. By using in vitro assays of neural crest and placodal cell differentiation, we now provide evidence that Wnt signals impose caudal character on neural plate border cells at the late gastrula stage, and that under these conditions, BMP signals induce neural crest instead of rostral placodal cells. We also provide evidence that both caudal neural and caudal neural plate border cells become independent of further exposure to Wnt signals at the head fold stage. Thus, the status of Wnt signaling in ectodermal cells at the late gastrula stage regulates the rostrocaudal patterning of both neural plate and neural plate border, providing a coordinated spatial and temporal control of the early development of the central and peripheral nervous systems. PMID- 18286181 TI - Isolation, cloning and structural characterisation of boophilin, a multifunctional Kunitz-type proteinase inhibitor from the cattle tick. AB - Inhibitors of coagulation factors from blood-feeding animals display a wide variety of structural motifs and inhibition mechanisms. We have isolated a novel inhibitor from the cattle tick Boophilus microplus, one of the most widespread parasites of farm animals. The inhibitor, which we have termed boophilin, has been cloned and overexpressed in Escherichia coli. Mature boophilin is composed of two canonical Kunitz-type domains, and inhibits not only the major procoagulant enzyme, thrombin, but in addition, and by contrast to all other previously characterised natural thrombin inhibitors, significantly interferes with the proteolytic activity of other serine proteinases such as trypsin and plasmin. The crystal structure of the bovine alpha-thrombin.boophilin complex, refined at 2.35 A resolution reveals a non-canonical binding mode to the proteinase. The N-terminal region of the mature inhibitor, Q16-R17-N18, binds in a parallel manner across the active site of the proteinase, with the guanidinium group of R17 anchored in the S(1) pocket, while the C-terminal Kunitz domain is negatively charged and docks into the basic exosite I of thrombin. This binding mode resembles the previously characterised thrombin inhibitor, ornithodorin which, unlike boophilin, is composed of two distorted Kunitz modules. Unexpectedly, both boophilin domains adopt markedly different orientations when compared to those of ornithodorin, in its complex with thrombin. The N-terminal boophilin domain rotates 9 degrees and is displaced by 6 A, while the C-terminal domain rotates almost 6 degrees accompanied by a 3 A displacement. The reactive site loop of the N-terminal Kunitz domain of boophilin with its P(1) residue, K31, is fully solvent exposed and could thus bind a second trypsin-like proteinase without sterical restraints. This finding explains the formation of a ternary thrombin.boophilin.trypsin complex, and suggests a mechanism for prothrombinase inhibition in vivo. PMID- 18286183 TI - Anaemia in acute HIV-1 subtype C infection. AB - BACKGROUND: The high prevalence of anaemia and the increased morbidity and mortality associated with anaemia during AIDS has been well described yet there has been little information about anaemia and changes in haemoglobin levels during acute and early HIV-1 infection. METHODS: HIV-negative women (n = 245) were enrolled into an observational cohort as part of the Centre for the AIDS Programme of Research in South Africa (CAPRISA) Acute Infection Study. Acute infection was diagnosed following a positive HIV RNA PCR in the absence of antibodies, or detection of HIV-1 antibodies within 3 months of a previously negative antibody test. Haemotologic parameters were assessed before infection and at regular intervals in the first twelve months of HIV infection. RESULTS: Fifty-seven participants with acute HIV infection were identified at a median of 14.5 days post-infection (range 10-81) and were enrolled in the CAPRISA Acute Infection cohort at a median of 41 days post-infection (range 15-104). Mean haemoglobin prior to HIV-1 infection was 12.7 g/dL, with a mean decline of 0.46 g/dL following infection. The prevalence of anaemia increased from 25.0% prior to HIV-1 infection to 52.6% at 3 months post-infection, 61.1% at 6 months post infection, and 51.4% at 12 months post-infection. CONCLUSIONS: Haematologic derangements and anaemia with a trend towards iron deficiency are common with acute HIV-1 subtype C infection in this small cohort. The negative impact of anaemia concurrent with established HIV infection upon morbidity and mortality has been well documented but the prognostic potential and long-term effects of anaemia during acute HIV-1 infection remain unknown. PMID- 18286184 TI - Malignant catarrhal fever induced by alcelaphine herpesvirus 1 is associated with proliferation of CD8+ T cells supporting a latent infection. AB - Alcelaphine herpesvirus 1 (AlHV-1), carried by wildebeest asymptomatically, causes malignant catarrhal fever (WD-MCF) when cross-species transmitted to a variety of susceptible species of the Artiodactyla order. Experimentally, WD-MCF can be induced in rabbits. The lesions observed are very similar to those described in natural host species. Here, we used the rabbit model and in vivo 5 Bromo-2'-Deoxyuridine (BrdU) incorporation to study WD-MCF pathogenesis. The results obtained can be summarized as follows. (i) AlHV-1 infection induces CD8(+) T cell proliferation detectable as early as 15 days post-inoculation. (ii) While the viral load in peripheral blood mononuclear cells remains below the detection level during most of the incubation period, it increases drastically few days before death. At that time, at least 10% of CD8(+ )cells carry the viral genome; while CD11b(+), IgM(+) and CD4(+) cells do not. (iii) RT-PCR analyses of mononuclear cells isolated from the spleen and the popliteal lymph node of infected rabbits revealed no expression of ORF25 and ORF9, low or no expression of ORF50, and high or no expression of ORF73. Based on these data, we propose a new model for the pathogenesis of WD-MCF. This model relies on proliferation of infected CD8(+) cells supporting a predominantly latent infection. PMID- 18286185 TI - Genetic analysis of the role of proteolysis in the activation of latent myostatin. AB - Myostatin is a secreted protein that normally acts to limit skeletal muscle growth. As a result, there is considerable interest in developing agents capable of blocking myostatin activity, as such agents could have widespread applications for the treatment of muscle degenerative and wasting conditions. Myostatin normally exists in an inactive state in which the mature C-terminal portion of the molecule is bound non-covalently to its N-terminal propeptide. We previously showed that this latent complex can be activated in vitro by cleavage of the propeptide with members of the bone morphogenetic protein-1/tolloid (BMP-1/TLD) family of metalloproteases. Here, I show that mice engineered to carry a germline point mutation rendering the propeptide protease-resistant exhibit increases in muscle mass approaching those seen in mice completely lacking myostatin. Mice homozygous for the point mutation have increased muscling even though their circulating levels of myostatin protein are dramatically increased, consistent with an inability of myostatin to be activated from its latent state. Furthermore, I show that a loss-of-function mutation in Tll2, which encodes one member of this protease family, has a small, but significant, effect on muscle mass, implying that its function is likely redundant with those of other family members. These findings provide genetic support for the hypothesis that proteolytic cleavage of the propeptide by BMP-1/TLD proteases plays a critical role in the activation of latent myostatin in vivo and suggest that targeting the activities of these proteases may be an effective therapeutic strategy for enhancing muscle growth in clinical settings of muscle loss and degeneration. PMID- 18286186 TI - Colouration and colour changes of the fiddler crab, Uca capricornis: a descriptive study. AB - Colour changes in animals may be triggered by a variety of social and environmental factors and may occur over a matter of seconds or months. Crustaceans, like fiddler crabs (genus Uca), are particularly adept at changing their colour and have been the focus of numerous studies. However, few of these studies have attempted to quantitatively describe the individual variation in colour and pattern or their adaptive significance. This paper quantitatively describes the colour patterns of the fiddler crab Uca capricornis and their ability to change on a socially significant timescale. The most dramatic changes in colour pattern are associated with moulting. These ontogenetic changes result in a general reduction of the colour pattern with increasing size, although females are more colourful and variable than similarly-sized males. Uca capricornis are also capable of rapid colour changes in response to stress, but show no endogenous rhythms associated with the semilunar and tidal cycles commonly reported in other fiddler crabs. The extreme colour polymorphism and the relative stability of the colour patterns in Uca capricornis are consistent with their use in visually mediated mate recognition. PMID- 18286187 TI - Safety and efficacy of methylene blue combined with artesunate or amodiaquine for uncomplicated falciparum malaria: a randomized controlled trial from Burkina Faso. AB - BACKGROUND: Besides existing artemisinin-based combination therapies, alternative safe, effective and affordable drug combinations against falciparum malaria are needed. Methylene blue (MB) was the first synthetic antimalarial drug ever used, and recent studies have been promising with regard to its revival in malaria therapy. The objective of this study was to assess the safety and efficacy of two MB-based malaria combination therapies, MB-artesunate (AS) and MB-amodiaquine (AQ), compared to the local standard of care, AS-AQ, in Burkina Faso. METHODS AND FINDINGS: Open-label randomised controlled phase II study in 180 children aged 6 10 years with uncomplicated falciparum malaria in Nouna, north-western Burkina Faso. Follow-up was for 28 days and analysis by intention-to-treat. The treatment groups were similar in baseline characteristics and there was only one loss to follow-up. No drug-related serious adverse events and no deaths occurred. MB containing regimens were associated with mild vomiting and dysuria. No early treatment failures were observed. Parasite clearance time differed significantly among groups and was the shortest with MB-AS. By day 14, the rates of adequate clinical and parasitological response after PCR-based correction for recrudescence were 87% for MB-AS, 100% for MB-AQ (p = 0.004), and 100% for AS-AQ (p = 0.003). By day 28, the respective figure was lowest for MB-AS (62%), intermediate for the standard treatment AS-AQ (82%; p = 0.015), and highest for MB-AQ (95%; p<0.001; p = 0.03). CONCLUSIONS: MB-AQ is a promising alternative drug combination against malaria in Africa. Moreover, MB has the potential to further accelerate the rapid parasite clearance of artemisinin-based combination therapies. More than a century after the antimalarial properties of MB had been described, its role in malaria control deserves closer attention. TRIAL REGISTRATION: ClinicalTrials.gov NCT00354380. PMID- 18286189 TI - Chromosomal instability in near-diploid colorectal cancer: a link between numbers and structure. AB - Chromosomal instability (CIN) plays a crucial role in tumor development and occurs mainly as the consequence of either missegregation of normal chromosomes (MSG) or structural rearrangement (SR). However, little is known about the respective chromosomal targets of MSG and SR and the way these processes combined within tumors to generate CIN. To address these questions, we karyotyped a consecutive series of 96 near-diploid colorectal cancers (CRCs) and distinguished chromosomal changes generated by either MSG or SR in tumor cells. Eighty-three tumors (86%) presented with chromosomal abnormalities that contained both MSGs and SRs to varying degrees whereas all 13 others (14%) showed normal karyotype. Using a maximum likelihood statistical method, chromosomes affected by MSG or SR and likely to represent changes that are selected for during tumor progression were found to be different and mostly mutually exclusive. MSGs and SRs were not randomly associated within tumors, delineating two major pathways of chromosome alterations that consisted of either chromosome gains by MSG or chromosomal losses by both MSG and SR. CRCs showing microsatellite instability (MSI) presented with either normal karyotype or chromosome gains whereas MSS (microsatellite stable) CRCs exhibited a combination of the two pathways. Taken together, these data provide new insights into the respective involvement of MSG and SR in near-diploid colorectal cancers, showing how these processes target distinct portions of the genome and result in specific patterns of chromosomal changes according to MSI status. PMID- 18286188 TI - Brain fatty acid binding protein (Fabp7) is diurnally regulated in astrocytes and hippocampal granule cell precursors in adult rodent brain. AB - Brain fatty acid binding protein (Fabp7), which is important in early nervous system development, is expressed in astrocytes and neuronal cell precursors in mature brain. We report here that levels of Fabp7 mRNA in adult murine brain change over a 24 hour period. Unlike Fabp5, a fatty acid binding protein that is expressed widely in various cell types within brain, RNA analysis revealed that Fabp7 mRNA levels were elevated during the light period and lower during dark in brain regions involved in sleep and activity mechanisms. This pattern of Fabp7 mRNA expression was confirmed using in situ hybridization and found to occur throughout the entire brain. Changes in the intracellular distribution of Fabp7 mRNA were also evident over a 24 hour period. Diurnal changes in Fabp7, however, were not found in postnatal day 6 brain, when astrocytes are not yet mature. In contrast, granule cell precursors of the subgranular zone of adult hippocampus did undergo diurnal changes in Fabp7 expression. These changes paralleled oscillations in Fabp7 mRNA throughout the brain suggesting that cell-coordinated signals likely control brain-wide Fabp7 mRNA expression. Immunoblots revealed that Fabp7 protein levels also underwent diurnal changes in abundance, with peak levels occurring in the dark period. Of clock or clock-regulated genes, the synchronized, global cycling pattern of Fabp7 expression is unique and implicates glial cells in the response or modulation of activity and/or circadian rhythms. PMID- 18286190 TI - Clocking the Lyme spirochete. AB - In order to clear the body of infecting spirochetes, phagocytic cells must be able to get hold of them. In real-time phase-contrast videomicroscopy we were able to measure the speed of Borrelia burgdorferi (Bb), the Lyme spirochete, moving back and forth across a platelet to which it was tethered. Its mean crossing speed was 1,636 microm/min (N = 28), maximum, 2800 microm/min (N = 3). This is the fastest speed recorded for a spirochete, and upward of two orders of magnitude above the speed of a human neutrophil, the fastest cell in the body. This alacrity and its interpretation, in an organism with bidirectional motor capacity, may well contribute to difficulties in spirochete clearance by the host. PMID- 18286191 TI - Recombination and population mosaic of a multifunctional viral gene, adeno associated virus cap. AB - Homologous recombination is a dominant force in evolution and results in genetic mosaics. To detect evidence of recombination events and assess the biological significance of genetic mosaics, genome sequences for various viral populations of reasonably large size are now available in the GenBank. We studied a multi functional viral gene, the adeno-associated virus (AAV) cap gene, which codes for three capsid proteins, VP1, VP2 and VP3. VP1-3 share a common C-terminal domain corresponding to VP3, which forms the viral core structure, while the VP1 unique N-terminal part contains an enzymatic domain with phospholipase A2 activity. Our recombinant detection program (RecI) revealed five novel recombination events, four of which have their cross-over points in the N-terminal, VP1 and VP2 unique region. Comparison of phylogenetic trees for different cap gene regions confirmed discordant phylogenies for the recombinant sequences. Furthermore, differences in the phylogenetic tree structures for the VP1 unique (VP1u) region and the rest of cap highlighted the mosaic nature of cap gene in the AAV population: two dominant forms of VP1u sequences were identified and these forms are linked to diverse sequences in the rest of cap gene. This observation together with the finding of frequent recombination in the VP1 and 2 unique regions suggests that this region is a recombination hot spot. Recombination events in this region preserve protein blocks of distinctive functions and contribute to convergence in VP1u and divergence of the rest of cap. Additionally the possible biological significance of two dominant VP1u forms is inferred. PMID- 18286192 TI - Revealing historic invasion patterns and potential invasion sites for two non native plant species. AB - The historical spatio-temporal distribution of invasive species is rarely documented, hampering efforts to understand invasion dynamics, especially at regional scales. Reconstructing historical invasions through use of herbarium records combined with spatial trend analysis and modeling can elucidate spreading patterns and identify susceptible habitats before invasion occurs. Two perennial species were chosen to contrast historic and potential phytogeographies: Japanese knotweed (Polygonum cuspidatum), introduced intentionally across the US; and mugwort (Artemisia vulgaris), introduced largely accidentally to coastal areas. Spatial analysis revealed that early in the invasion, both species have a stochastic distribution across the contiguous US, but east of the 90(th) meridian, which approximates the Mississippi River, quickly spread to adjacent counties in subsequent decades. In contrast, in locations west of the 90(th) meridian, many populations never spread outside the founding county, probably a result of encountering unfavorable environmental conditions. Regression analysis using variables categorized as environmental or anthropogenic accounted for 24% (Japanese knotweed) and 30% (mugwort) of the variation in the current distribution of each species. Results show very few counties with high habitat suitability (>/=80%) remain un-invaded (5 for Japanese knotweed and 6 for mugwort), suggesting these perennials are reaching the limits of large-scale expansion. Despite differences in initial introduction loci and pathways, Japanese knotweed and mugwort demonstrate similar historic patterns of spread and show declining rates of regional expansion. Invasion mitigation efforts should be concentrated on areas identified as highly susceptible that border invaded regions, as both species demonstrate secondary expansion from introduction loci. PMID- 18286193 TI - Measuring the meltdown: drivers of global amphibian extinction and decline. AB - Habitat loss, climate change, over-exploitation, disease and other factors have been hypothesised in the global decline of amphibian biodiversity. However, the relative importance of and synergies among different drivers are still poorly understood. We present the largest global analysis of roughly 45% of known amphibians (2,583 species) to quantify the influences of life history, climate, human density and habitat loss on declines and extinction risk. Multi-model Bayesian inference reveals that large amphibian species with small geographic range and pronounced seasonality in temperature and precipitation are most likely to be Red-Listed by IUCN. Elevated habitat loss and human densities are also correlated with high threat risk. Range size, habitat loss and more extreme seasonality in precipitation contributed to decline risk in the 2,454 species that declined between 1980 and 2004, compared to species that were stable (n = 1,545) or had increased (n = 28). These empirical results show that amphibian species with restricted ranges should be urgently targeted for conservation. PMID- 18286194 TI - Recombination-mediated genetic engineering of a bacterial artificial chromosome clone of modified vaccinia virus Ankara (MVA). AB - The production, manipulation and rescue of a bacterial artificial chromosome clone of Vaccinia virus (VAC-BAC) in order to expedite construction of expression vectors and mutagenesis of the genome has been described (Domi & Moss, 2002, PNAS99 12415-20). The genomic BAC clone was 'rescued' back to infectious virus using a Fowlpox virus helper to supply transcriptional machinery. We apply here a similar approach to the attenuated strain Modified Vaccinia virus Ankara (MVA), now widely used as a safe non-replicating recombinant vaccine vector in mammals, including humans. Four apparently full-length, rescuable clones were obtained, which had indistinguishable immunogenicity in mice. One clone was shotgun sequenced and found to be identical to the parent. We employed GalK recombination mediated genetic engineering (recombineering) of MVA-BAC to delete five selected viral genes. Deletion of C12L, A44L, A46R or B7R did not significantly affect CD8(+) T cell immunogenicity in BALB/c mice, but deletion of B15R enhanced specific CD8(+) T cell responses to one of two endogenous viral epitopes (from the E2 and F2 proteins), in accordance with published work (Staib et al., 2005, J. Gen. Virol.86, 1997-2006). In addition, we found a higher frequency of triple positive IFN-gamma, TNF-alpha and IL-2 secreting E3-specific CD8+ T-cells 8 weeks after vaccination with MVA lacking B15R. Furthermore, a recombinant vaccine capable of inducing CD8(+) T cells against an epitope from Plasmodium berghei was created using GalK counterselection to insert an antigen expression cassette lacking a tandem marker gene into the traditional thymidine kinase locus of MVA BAC. MVA continues to feature prominently in clinical trials of recombinant vaccines against diseases such as HIV-AIDS, malaria and tuberculosis. Here we demonstrate in proof-of-concept experiments that MVA-BAC recombineering is a viable route to more rapid and efficient generation of new candidate mutant and recombinant vaccines based on a clinically deployable viral vector. PMID- 18286195 TI - Loss of cytokine-STAT5 signaling in the CNS and pituitary gland alters energy balance and leads to obesity. AB - Signal transducers and activators of transcription (STATs) are critical components of cytokine signaling pathways. STAT5A and STAT5B (STAT5), the most promiscuous members of this family, are highly expressed in specific populations of hypothalamic neurons in regions known to mediate the actions of cytokines in the regulation of energy balance. To test the hypothesis that STAT5 signaling is essential to energy homeostasis, we used Cre-mediated recombination to delete the Stat5 locus in the CNS. Mutant males and females developed severe obesity with hyperphagia, impaired thermal regulation in response to cold, hyperleptinemia and insulin resistance. Furthermore, central administration of GM-CSF mediated the nuclear accumulation of STAT5 in hypothalamic neurons and reduced food intake in control but not in mutant mice. These results demonstrate that STAT5 mediates energy homeostasis in response to endogenous cytokines such as GM-CSF. PMID- 18286196 TI - Brain neuronal CB2 cannabinoid receptors in drug abuse and depression: from mice to human subjects. AB - BACKGROUND: Addiction and major depression are mental health problems associated with stressful events in life with high relapse and reoccurrence even after treatment. Many laboratories were not able to detect the presence of cannabinoid CB2 receptors (CB2-Rs) in healthy brains, but there has been demonstration of CB2 R expression in rat microglial cells and other brain associated cells during inflammation. Therefore, neuronal expression of CB2-Rs had been ambiguous and controversial and its role in depression and substance abuse is unknown. METHODOLOGY/PRINCIPAL FINDINGS: In this study we tested the hypothesis that genetic variants of CB2 gene might be associated with depression in a human population and that alteration in CB2 gene expression may be involved in the effects of abused substances including opiates, cocaine and ethanol in rodents. Here we demonstrate that a high incidence of (Q63R) but not (H316Y) polymorphism in the CB2 gene was found in Japanese depressed subjects. CB2-Rs and their gene transcripts are expressed in the brains of naive mice and are modulated following exposure to stressors and administration of abused drugs. Mice that developed alcohol preference had reduced CB2 gene expression and chronic treatment with JWH015 a putative CB2-R agonist, enhanced alcohol consumption in stressed but not in control mice. The direct intracerebroventricular microinjection of CB2 anti sense oligonucleotide into the mouse brain reduced mouse aversions in the plus maze test, indicating the functional presence of CB2-Rs in the brain that modifies behavior. We report for the using electron microscopy the sub cellular localization of CB2-Rs that are mainly on post-synaptic elements in rodent brain. CONCLUSIONS/SIGNIFICANCE: Our data demonstrate the functional expression of CB2 Rs in brain that may provide novel targets for the effects of cannabinoids in depression and substance abuse disorders beyond neuro-immunocannabinoid activity. PMID- 18286197 TI - Human VRK1 is an early response gene and its loss causes a block in cell cycle progression. AB - BACKGROUND: In mammalian cells regulatory proteins controlling the cell cycle are necessary due to the requirements of living in a heterogeneous environment of cell-interactions and growth factors. VRK1 is a novel serine-threonine kinase that phosphorylates several transcription factors and is associated with proliferation phenotypes. METHODOLOGY/PRINCIPAL FINDINGS: In this report VRK1 has been identified as regulated in the cell cycle. VRK1 gene expression is activated by the addition of serum to starved cells, indicating it is required for the exit of G0 phase and entry in G1; a response that parallels the re-expression of MYC, FOS and CCND1 (cyclin D1) genes, suggesting that VRK1 is an early-response gene. VRK1 gene expression is also shutdown by serum withdrawal. The human VRK1 gene promoter cloned in a luciferase reporter responds similarly to serum. In response to serum, the level of VRK1 protein expression has a positive correlation with cell proliferation markers such as phosphorylated-Rb or PCNA, and is inversely correlated with cell cycle inhibitors such as p27. The elimination of VRK1 by siRNA results in a G1 block in cell division, and in loss of phosphorylated-Rb, cyclin D1, and other proliferation markers. Elimination of VRK1 by siRNA induces a reduction of cell proliferation. VRK1 colocalizes with p63 in proliferating areas of squamous epithelium, and identifies a subpopulation in the basal layer. CONCLUSIONS/SIGNIFICANCE: VRK1 is an immediate early response gene required for entry in G1, and due to its implication in normal cell proliferation and division, might be a new target for development of inhibitors of cellular proliferation. PMID- 18286198 TI - A cross-talk between TrkB and Ret tyrosine kinases receptors mediates neuroblastoma cells differentiation. AB - Understanding the interplay between intracellular signals initiated by multiple receptor tyrosine kinases (RTKs) to give the final cell phenotype is a major pharmacological challenge. Retinoic acid (RA)-treatment of neuroblastoma (NB) cells implicates activation of Ret and TrkB RTKs as critical step to induce cell differentiation. By studying the signaling interplay between TrkB and Ret as paradigmatic example, here we demonstrate the existence of a cross-talk mechanism between the two unrelated receptors that is needed to induce the cell differentiation. Indeed, we show that TrkB receptor promotes Ret phosphorylation by a mechanism that does not require GDNF. This reveals to be a key mechanism, since blocking either TrkB or Ret by small interfering RNA causes a failure in NB biochemical and morphological differentiation. Our results provide the first evidence that a functional transactivation between distinct tyrosine kinases receptors is required for an important physiological process. PMID- 18286199 TI - Adherent self-renewable human embryonic stem cell-derived neural stem cell line: functional engraftment in experimental stroke model. AB - BACKGROUND: Human embryonic stem cells (hESCs) offer a virtually unlimited source of neural cells for structural repair in neurological disorders, such as stroke. Neural cells can be derived from hESCs either by direct enrichment, or by isolating specific growth factor-responsive and expandable populations of human neural stem cells (hNSCs). Studies have indicated that the direct enrichment method generates a heterogeneous population of cells that may contain residual undifferentiated stem cells that could lead to tumor formation in vivo. METHODS/PRINCIPAL FINDINGS: We isolated an expandable and homogenous population of hNSCs (named SD56) from hESCs using a defined media supplemented with epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and leukemia inhibitory growth factor (LIF). These hNSCs grew as an adherent monolayer culture. They were fully neuralized and uniformly expressed molecular features of NSCs, including nestin, vimentin and radial glial markers. These hNSCs did not express the pluripotency markers Oct4 or Nanog, nor did they express markers for the mesoderm or endoderm lineages. The self-renewal property of the hNSCs was characterized by a predominant symmetrical mode of cell division. The SD56 hNSCs differentiated into neurons, astrocytes and oligodendrocytes throughout multiple passages in vitro, as well as after transplantation. Together, these criteria confirm the definitive NSC identity of the SD56 cell line. Importantly, they exhibited no chromosome abnormalities and did not form tumors after implantation into rat ischemic brains and into naive nude rat brains and flanks. Furthermore, hNSCs isolated under these conditions migrated toward the ischemia-injured adult brain parenchyma and improved the independent use of the stroke-impaired forelimb two months post-transplantation. CONCLUSIONS/SIGNIFICANCE: The SD56 human neural stem cells derived under the reported conditions are stable, do not form tumors in vivo and enable functional recovery after stroke. These properties indicate that this hNSC line may offer a renewable, homogenous source of neural cells that will be valuable for basic and translational research. PMID- 18286200 TI - 18-month effectiveness of short-course antiretroviral regimens combined with alternatives to breastfeeding to prevent HIV mother-to-child transmission. AB - OBJECTIVE: We assessed the 18-month effectiveness of short-course (sc) antiretroviral peripartum regimens combined with alternatives to prolonged breastfeeding to prevent mother-to-child transmission (MTCT) of HIV-1 in Abidjan, Cote d'Ivoire. METHODOLOGY: HIV-1 infected pregnant women received from >/=32-36 weeks of gestation scZidovudine (ZDV)+/-Lamivudine (3TC)+single-dose Nevirapine (sdNVP) at delivery within the ANRS 1201/1202 DITRAME-Plus cohort (2001-2003). Neonates received a sdNVP+7-day ZDV prophylaxis. Two infant-feeding interventions were systematically offered free of charge: formula-feeding or exclusive shortened breastfeeding with early cessation from four months. The reference group was the ANRS 049a DITRAME cohort (1994-2000) exposed to scZDV from 36 weeks, then to prolonged breastfeeding. Pediatric HIV infection was defined by a positive plasma HIV-1 RNA at any age, or if aged >/=18 months, a positive HIV-1 serology. Turnbull estimates of cumulative transmission risks (CTR) and effectiveness (HIV-free survival) were compared by exposure group using a Cox model. FINDINGS: Among 926 live-born children enrolled, 107 (11.6%) were HIV infected at 18 months. CTRs were 22.3% (95% confidence interval[CI]:16-30%) in the 238 ZDV long-term breastfed reference group, 15.9% (CI:10-27%) in the 169 ZDV+sdNVP shortened breastfed group; 9.4% (CI:6-14%) in the 195 ZDV+sdNVP formula fed group; 6.8% (CI:4-11%) in the 198 ZDV+3TC+sdNVP shortened breastfed group, and 5.6% (CI:2-10%) in the 126 ZDV+3TC+sdNVP formula-fed group. Each combination had a significantly higher effectiveness than the ZDV long-term breastfed group except for ZDV+sdNVP shortened breastfed children, ranging from 51% (CI:20-70%) for ZDV+sdNVP formula fed children to 63% (CI:40-80%) for ZDV+3TC+NVPsd shortened breastfed children, after adjustment for maternal eligibility for antiretroviral therapy (ART), home delivery and low birth-weight. Substantial MTCT risk reductions are reachable in Africa, even in short-term breastfed children. The two sc antiretroviral combinations associated to any of the two infant feeding interventions, formula-feeding and shortened breastfeeding, reduce significantly MTCT with long-term benefit until age 18 months and without increasing mortality. PMID- 18286201 TI - Use of cDNA tiling arrays for identifying protein interactions selected by in vitro display technologies. AB - In vitro display technologies such as mRNA display are powerful screening tools for protein interaction analysis, but the final cloning and sequencing processes represent a bottleneck, resulting in many false negatives. Here we describe an application of tiling array technology to identify specifically binding proteins selected with the in vitro virus (IVV) mRNA display technology. We constructed transcription-factor tiling (TFT) arrays containing approximately 1,600 open reading frame sequences of known and predicted mouse transcription-regulatory factors (334,372 oligonucleotides, 50-mer in length) to analyze cDNA fragments from mRNA-display screening for Jun-associated proteins. The use of the TFT arrays greatly increased the coverage of known Jun-interactors to 28% (from 14% with the cloning and sequencing approach), without reducing the accuracy ( approximately 75%). This method could detect even targets with extremely low expression levels (less than a single mRNA copy per cell in whole brain tissue). This highly sensitive and reliable method should be useful for high-throughput protein interaction analysis on a genome-wide scale. PMID- 18286202 TI - The role of IRE1alpha in the degradation of insulin mRNA in pancreatic beta cells. AB - BACKGROUND: The endoplasmic reticulum (ER) is a cellular compartment for the biosynthesis and folding of newly synthesized secretory proteins such as insulin. Perturbations to ER homeostasis cause ER stress and subsequently activate cell signaling pathways, collectively known as the Unfolded Protein Response (UPR). IRE1alpha is a central component of the UPR. In pancreatic beta-cells, IRE1alpha also functions in the regulation of insulin biosynthesis. PRINCIPAL FINDINGS: Here we report that hyperactivation of IRE1alpha caused by chronic high glucose treatment or IRE1alpha overexpression leads to insulin mRNA degradation in pancreatic beta-cells. Inhibition of IRE1alpha signaling using its dominant negative form prevents insulin mRNA degradation. Islets from mice heterozygous for IRE1alpha retain expression of more insulin mRNA after chronic high glucose treatment than do their wild-type littermates. CONCLUSIONS/SIGNIFICANCE: These results reveal a role of IRE1alpha in insulin mRNA expression under ER stress conditions caused by chronic high glucose. The rapid degradation of insulin mRNA could provide immediate relief for the ER and free up the translocation machinery. Thus, this mechanism would preserve ER homeostasis and help ensure that the insulin already inside the ER can be properly folded and secreted. This adaptation may be crucial for the maintenance of beta-cell homeostasis and may explain why the beta-cells of type 2 diabetic patients with chronic hyperglycemia stop producing insulin in the absence of apoptosis. This mechanism may also be involved in suppression of the autoimmune type 1 diabetes by reducing the amount of misfolded insulin, which could be a source of "neo-autoantigens." PMID- 18286203 TI - Cross-species functionality of pararetroviral elements driving ribosome shunting. AB - BACKGROUND: Cauliflower mosaic virus (CaMV) and Rice tungro bacilliform virus (RTBV) belong to distinct genera of pararetroviruses infecting dicot and monocot plants, respectively. In both viruses, polycistronic translation of pregenomic (pg) RNA is initiated by shunting ribosomes that bypass a large region of the pgRNA leader with several short (s)ORFs and a stable stem-loop structure. The shunt requires translation of a 5'-proximal sORF terminating near the stem. In CaMV, mutations knocking out this sORF nearly abolish shunting and virus viability. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that two distant regions of the CaMV leader that form a minimal shunt configuration comprising the sORF, a bottom part of the stem, and a shunt landing sequence can be replaced by heterologous sequences that form a structurally similar configuration in RTBV without any dramatic effect on shunt-mediated translation and CaMV infectivity. The CaMV-RTBV chimeric leader sequence was largely stable over five viral passages in turnip plants: a few alterations that did eventually occur in the virus progenies are indicative of fine tuning of the chimeric sequence during adaptation to a new host. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate cross-species functionality of pararetroviral cis-elements driving ribosome shunting and evolutionary conservation of the shunt mechanism. We are grateful to Matthias Muller and Sandra Pauli for technical assistance. This work was initiated at Friedrich Miescher Institute (Basel, Switzerland). We thank Prof. Thomas Boller for hosting the group at the Institute of Botany. PMID- 18286204 TI - Precancerous stem cells can serve as tumor vasculogenic progenitors. AB - Tumor neo-vascularization is critical for tumor growth, invasion and metastasis, which has been considered to be mediated by a mechanism of angiogenesis. However, histopathological studies have suggested that tumor cells might be the progenitor for tumor vasculature. Recently, we have reported that the precancerous stem cells (pCSCs) representing the early stage of developing cancer stem cells (CSCs), have the potential for both benign and malignant differentiation. Therefore, we investigated whether pCSCs serve as progenitors for tumor vasculogenesis. Herein, we report that in the pCSC-derived tumors, most blood vessels were derived from pCSCs. Some pCSCs constitutively expressed vasculogenic receptor VEGFR-2, which can be up-regulated by hypoxia and angiogenesis-promoting cytokines, such as GM-CSF, Flt3 ligand, and IL-13. The pCSCs are much more potent in tumor vasculogenesis than the differentiated tumor monocytic cells (TMCs) from the same tumor, which had comparable or even higher capacity to produce some vascular growth factors, suggesting that the potent tumor vasculogenesis of pCSCs is associated with their intrinsic stem-like property. Consistently tumor vasculogenesis was also observed in human cancers such as cervical cancer and breast cancer and xenograft lymphoma. Our studies indicate that pCSCs can serve as tumor vasculogenic stem/progenitor cells (TVPCs), and may explain why anti angiogenic cancer therapy trials are facing challenge. PMID- 18286205 TI - The enhancer of trithorax and polycomb corto interacts with cyclin G in Drosophila. AB - BACKGROUND: Polycomb (PcG) and trithorax (trxG) genes encode proteins involved in the maintenance of gene expression patterns, notably Hox genes, throughout development. PcG proteins are required for long-term gene repression whereas TrxG proteins are positive regulators that counteract PcG action. PcG and TrxG proteins form large complexes that bind chromatin at overlapping sites called Polycomb and Trithorax Response Elements (PRE/TRE). A third class of proteins, so called "Enhancers of Trithorax and Polycomb" (ETP), interacts with either complexes, behaving sometimes as repressors and sometimes as activators. The role of ETP proteins is largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: In a two hybrid screen, we identified Cyclin G (CycG) as a partner of the Drosophila ETP Corto. Inactivation of CycG by RNA interference highlights its essential role during development. We show here that Corto and CycG directly interact and bind to each other in embryos and S2 cells. Moreover, CycG is targeted to polytene chromosomes where it co-localizes at multiple sites with Corto and with the PcG factor Polyhomeotic (PH). We observed that corto is involved in maintaining Abd-B repression outside its normal expression domain in embryos. This could be achieved by association between Corto and CycG since both proteins bind the regulatory element iab-7 PRE and the promoter of the Abd-B gene. CONCLUSIONS/SIGNIFICANCE: Our results suggest that CycG could regulate the activity of Corto at chromatin and thus be involved in changing Corto from an Enhancer of TrxG into an Enhancer of PcG. PMID- 18286206 TI - Modeling transcriptome based on transcript-sampling data. AB - BACKGROUND: Newly-evolved multiplex sequencing technology has been bringing transcriptome sequencing into an unprecedented depth. Millions of transcript tags now can be acquired in a single experiment through parallelization. The significant increase in throughput and reduction in cost required us to address some fundamental questions, such as how many transcript tags do we have to sequence for a given transcriptome? How could we estimate the total number of unique transcripts for different cell types (transcriptome diversity) and the distribution of their copy numbers (transcriptome dynamics)? What is the probability that a transcript with a given expression level to be detected at a certain sampling depth? METHODOLOGY/PRINCIPAL FINDINGS: We developed a statistical model to evaluate these parameters based on transcriptome-sampling data. Three mixture models were exploited for their potentials to model the sampling frequencies. We demonstrated that relative abundances of all transcripts in a transcriptome follow the generalized inverse Gaussian distribution. The widely known beta and gamma distributions failed to fulfill the singular characteristics of relative abundance distribution, i.e., highly skewed toward zero and with a long tail. An estimator of transcriptome diversity and an analytical form of sampling growth curve were proposed in a coherent framework. Experimental data fitted this model very well and Monte Carlo simulations based on this model replicated sampling experiments in a remarkable precision. CONCLUSIONS: Taking human embryonic stem cell as a prototype, we demonstrated that sequencing tens of thousands of transcript tags in an ordinary EST/SAGE experiment was far from sufficient. In order to fully characterize a human transcriptome, millions of transcript tags had to be sequenced. This model lays a statistical basis for transcriptome-sampling experiments and in essence can be used in all sampling-based data. PMID- 18286207 TI - Modulation of interleukin-1 transcriptional response by the interaction between VRK2 and the JIP1 scaffold protein. AB - BACKGROUND: Cellular biological responses to specific stimulation are determined by a balance among signaling pathways. Protein interactions are likely to modulate these pathways. Vaccinia-related kinase-2 (VRK2) is a novel human kinase that can modulate different signaling pathways. PRINCIPAL FINDINGS: We report that in vivo, the activity of JIP1-JNK complexes is downregulated by VRK2 in response to interleukin-1beta. Also the reduction of endogenous VRK2 with shRNA increases the transcriptional response to IL-1beta. The JIP1 scaffold protein assembles three consecutive members of a given MAPK pathway forming signaling complexes and their signal can be modulated by interactions with regulatory proteins that remain to be identified. Knocking-down JIP1 with siRNA resulted in elimination of the AP1 transcriptional response to IL-1beta. VRK2, a member of novel Ser-Thr kinase family, is able to stably interact with JIP1, TAK1 and MKK7, but not JNK, and can be isolated forming oligomeric complexes with different proportions of TAK1, MKK7beta1 and JNK. JIP1 assembles all these proteins in an oligomeric signalosome. VRK2 binding to the JIP1 signalosome prevents the association of JNK and results in a reduction in its phosphorylation and downregulation of AP1-dependent transcription. CONCLUSIONS/SIGNIFICANCE: This work suggests that the intracellular level of VRK2 protein can modulate the flow through a signaling pathway and alter the response from a receptor that can be distributed by more than one pathway, and thus contribute to the cellular specificity of the response by forming alternative signaling complexes. Furthermore, the effect might be more general and affect other signaling routes assembled on the JIP1 scaffold protein for which a model is proposed. PMID- 18286208 TI - Maintenance of long-range DNA interactions after inhibition of ongoing RNA polymerase II transcription. AB - A relationship exists between nuclear architecture and gene activity and it has been proposed that the activity of ongoing RNA polymerase II transcription determines genome organization in the mammalian cell nucleus. Recently developed 3C and 4C technology allowed us to test the importance of transcription for nuclear architecture. We demonstrate that upon transcription inhibition binding of RNA polymerase II to gene regulatory elements is severely reduced. However, contacts between regulatory DNA elements and genes in the beta-globin locus are unaffected and the locus still interacts with the same genomic regions elsewhere on the chromosome. This is a general phenomenon since the great majority of intra and interchromosomal interactions with the ubiquitously expressed Rad23a gene are also not affected. Our data demonstrate that without transcription the organization and modification of nucleosomes at active loci and the local binding of specific trans-acting factors is unaltered. We propose that these parameters, more than transcription or RNA polymerase II binding, determine the maintenance of long-range DNA interactions. PMID- 18286209 TI - Recent advances in andrology-related stem cell research. AB - Stem cells hold great promise for regenerative medicine because of their ability to self-renew and to differentiate into various cell types. Although embryonic stem cells (BSC) have greater differentiation potential than adult stem cells, the former is lagging in reaching clinical applications because of ethical concerns and governmental restrictions. Bone marrow stem cells (BMSC) are the best-studied adult stem cells (ASC) and have the potential to treat a wide variety of diseases, including erectile dysfunction (ED) and male infertility. More recently discovered adipose tissue-derived stem cells (ADSC) are virtually identical to bone marrow stem cells in differentiation and therapeutic potential, but are easier and safer to obtain, can be harvested in larger quantities, and have the associated benefit of reducing obesity. Therefore, ADSC appear to be a better choice for future clinical applications. We have previously shown that ESC could restore the erectile function of neurogenic ED in rats, and we now have evidence that ADSC could do so as well. We are also investigating whether ADSC can differentiate into Leydig, Sertoli and male germ cells. The eventual goal is to use ADSC to treat male infertility and testosterone deficiency. PMID- 18286210 TI - Mutations of t-complex testis expressed gene 5 transcripts in the testis of sterile t-haplotype mutant mouse. AB - AIM: To determine the possible roles of the t-complex testis expressed gene 5 (Tctex5) on sperm functions, the full-length sequence of mRNA was studied and compared in the testis between the normal wild-type and the sterile t-haplotype mutant mice. METHODS: We applied rapid amplification of cDNA ends, Northern blot and reverse transcription polymerase chain reaction to analyze the full length of Tctex5 mRNAs isolated from testes of the wild-type and the t-haplotype mice. Reverse transcription polymerase chain reaction was used to semi-quantitatively compare expression of Tctex5 transcripts in the 16 tissues and 9.5 day stage embryos in the wild-type mice. E-translation was applied to estimate the amino acid sequences. RESULTS: One long and one short transcript of Tctex5 mRNA were discovered in mouse testis of wild-type (Tctex5(long-+) and Tctex5(short-+)) and t-haplotype (Tctex5(long-t) and Tctex5(short-t)) mice, respectively. Being enhanced only in the testis, Tctex5(long-t) had 17 point mutations and one 15-bp deletion in the exon 1 region, comparing with the Tctex5(long-+), whereas the Tctex5(short-t) was similar to the Tctex5(short-+). The short isoforms of Tctex5 mRNAs in the two models encoded exactly the same peptides, but the long isoforms did not. The estimated peptide encoded by Tctex5(long-t) had significant mutations on putative sites of phosphorylation and PP1 binding. CONCLUSION: We established that mutations that occur in the Tctex5 long transcript of the t haplotype mice are important for normal sperm function, whereas the short transcript of Tctex5 might have a conserved function among different tissues. PMID- 18286211 TI - Effects of estradiol-17beta and bisphenol A administered chronically to mice throughout pregnancy and lactation on the male pups' reproductive system. AB - AIM: To assess the effect of estradiol-17beta (E(2)) and bisphenol A (BPA) administered chronically by implanting a silicone tube throughout pregnancy and lactation on male pups' reproductive system in ICR mice. METHODS: Female mice were implanted with a tube filled with 10 ng, 500 ng, 1 microg, or 10 microg of E(2), or 100 microg or 5 mg of BPA, before mating. The tube was kept in the mice throughout pregnancy and lactation, until the pups had weaned at 4 weeks of age. During the period, E(2) was released from the tube at 120 pg or 6, 12 or 120 ng/day, and BPA at 1.2 or 60 microg/day. RESULTS: Most of the mice given 1 microg and 10 microg of E(2) did not maintain their pregnancy. However, the other groups showed high rates of birth, more than 70%. At age of 4 weeks, the male pups were killed. Body weight and reproductive organ weights (testes, epididymides and accessory reproductive glands) in the treated groups did not differ from the control values, whereas the percentage of seminiferous tubules in the testis with mature spermatids was significantly lower in the groups given 10 ng and 500 ng of E(2) and 5 mg of BPA than that in the control. CONCLUSION: Chronic exposure to E(2) and BPA might disrupt spermatogenesis in male pups. PMID- 18286215 TI - [Urology between Skylla and Charybdis]. PMID- 18286216 TI - Putting the person back into psychopathology: an intervention to reduce mental illness stigma in the classroom. AB - OBJECTIVE: This research aims to compare the effectiveness of two methods of teaching psychopathology in reducing stigma toward mental illness. Based on previous stigma research, a first-person, narrative approach was contrasted with traditional, diagnosis-centered education. STUDY 1 METHODS: Participants consisted of 53 undergraduates at a small, public university enrolled in two introductory psychology classes. During six hours of class time focused on psychopathology, one class received the experimental pedagogy while the other served as a control, receiving traditional instruction. Stigma was assessed pre- and post-intervention using a social distance scale and vignette design. Statistical analyses compared means and change scores between the two classes. STUDY 1 RESULTS: Students in the experimental classroom showed a significant decrease in stigma following the intervention, whereas those in the control group showed no change. STUDY 2 METHODS: A follow-up study was conducted to replicate the promising effects demonstrated in Study 1. Two additional classrooms (n = 48) were both exposed to the first-person, narrative pedagogy, and their stigma monitored pre- and post- intervention. STUDY 2 RESULTS: Students reported a significant decrease in stigma following the intervention. CONCLUSIONS: Together, these studies suggest that traditional methods of teaching psychopathology do not lessen mental illness stigma, a serious concern that can potentially be reconciled by incorporating more person-centered instructional methods. Results are discussed in terms of their implications for the way psychopathology is taught throughout the mental health field, as well as the practical application of stigma interventions woven into the curriculum. PMID- 18286218 TI - Effect of calcium supplementation on hip fractures. AB - There have been numerous studies of the effects of calcium supplementation, with or without vitamin D, on fractures. Individually, they have not provided clarity regarding calcium's anti-fracture efficacy, though they have established that calcium does have beneficial effects on bone density throughout the skeleton in women. Meta-analysis of these data suggests that total fracture numbers are diminished. However, the data from the 5,500 women involved in trials of calcium monotherapy show consistent adverse trends in numbers of hip fractures (relative risk 1.50, 95% CI 1.06-2.12). Observational data from the Study of Osteoporotic Fractures show a similar increase in risk of hip fracture associated with calcium use. We hypothesize that reduced periosteal expansion in women using calcium supplementation might account for the differences in anti-fracture efficacy of calcium at the hip, in comparison with other sites. Until there are further trial results to clarify this area, the present findings suggest that reliance on high calcium intakes to reduce the risk of hip fracture in older women is not appropriate. In addition, those at risk should be looking to other agents with a proven capacity to prevent hip fractures, such as bisphosphonates. PMID- 18286217 TI - Osteoporosis treatment and fracture incidence: the ICARO longitudinal study. AB - None of the available osteoporosis therapies completely abolish the risk of fracture. Among 862 patients on treatment with antiresorptive agents (alendronate, risedronate, and raloxifene) for >1 year a fragility fracture was observed in 9.5%/year. This incidence is considerably higher than that observed in randomized clinical trials. INTRODUCTION: Available osteoporosis therapies reduced in randomized controlled trials (RCTs) the risk of fracture by 30-50%. The proportion of patients suffering from new fractures while on active treatment ("inadequate clinical treatment response" or ICR) can be derived from the data of the RCTs, where confounding factors are usually controlled by the exclusion criteria. In the retrospective part of the ICARO study we observed a 8.9% annual incidence of ICR. Here we report the results of the longitudinal part of the study. METHODS: The study includes 862 women with severe postmenopausal osteoporosis. Ninety-two of these patients (10.7%) were defined as having ICR (9.5%/year) during therapy with antiresorptive drugs (alendronate, risedronate, and raloxifene) for at least 1 year. RESULTS: The ICR patients were comparable to patients who did not sustain clinical fractures with regard to body mass index, follow-up duration, number of prevalent vertebral fractures, type of osteoporosis treatment, proportion of patients taking calcium and vitamin D supplements, and compliance with treatment. Those with ICR were significantly older (p=0.032) and more frequently had multiple vertebral deformities (p=0.013). CONCLUSIONS: The incidence of ICR during treatment with antiresorptive agents among patients with severe postmenopausal osteoporosis in a routine setting is considerably higher than that observed in randomized clinical trials. PMID- 18286219 TI - Bone degeneration and recovery after early and late bisphosphonate treatment of ovariectomized wistar rats assessed by in vivo micro-computed tomography. AB - Bisphosphonates are antiresorptive drugs commonly used to treat osteoporosis. It is not clear, however, what the influence of the time point of treatment is. Recently developed in vivo micro-computed tomographic (CT) scanners offer the possibility to study such effects on bone microstructure in rats. The aim of this study was to determine the influence of early and late zoledronic acid treatment on bone in ovariectomized rats, using in vivo micro-CT. Twenty-nine female Wistar rats were divided into the following groups: ovariectomy (OVX, n = 5), OVX and zoledronic acid (ZOL) at week 0 (n = 8), OVX and ZOL at week 8 (n = 7), and sham (n = 9). CT scans were made of the proximal tibia at weeks 0, 2, 4, 8, 12, and 16; and bone structural parameters were determined in the metaphysis. Two fluorescent labels were administered to calculate dynamic histomorphometric parameters. At week 16, all groups were significantly different from each other in bone volume fraction (BV/TV), connectivity density, and trabecular number (Tb.N), except for the early ZOL and control groups which were not significantly different for any structural parameter. After ZOL treatment at week 8, BV/TV, structure model index, Tb.N, and trabecular thickness significantly improved in the late ZOL group. The OVX and ZOL groups showed, respectively, higher and lower bone formation rates than the control group. Early ZOL treatment inhibited all bone microstructural changes seen after OVX. Late ZOL treatment significantly improved bone microstructure, although the structure did not recover to original levels. Early ZOL treatment resulted in a significantly better microstructure than late treatment. However, late treatment was still significantly better than no treatment. PMID- 18286220 TI - Codon bias is a major factor explaining phage evolution in translationally biased hosts. AB - The size and diversity of bacteriophage populations require methodologies to quantitatively study the landscape of phage differences. Statistical approaches are confronted with small genome sizes forbidding significant single-phage analysis, and comparative methods analyzing full phage genomes represent an alternative but they are of difficult interpretation due to lateral gene transfer, which creates a mosaic spectrum of related phage species. Based on a large-scale codon bias analysis of 116 DNA phages hosted by 11 translationally biased bacteria belonging to different phylogenetic families, we observe that phage genomes are almost always under codon selective pressure imposed by translationally biased hosts, and we propose a classification of phages with translationally biased hosts which is based on adaptation patterns. We introduce a computational method for comparing phages sharing homologous proteins, possibly accepted by different hosts. We observe that throughout phages, independently from the host, capsid genes appear to be the most affected by host translational bias. For coliphages, genes involved in virion morphogenesis, host interaction and ssDNA binding are also affected by adaptive pressure. Adaptation affects long and small phages in a significant way. We analyze in more detail the Microviridae phage space to illustrate the potentiality of the approach. The small number of directions in adaptation observed in phages grouped around phi X174 is discussed in the light of functional bias. The adaptation analysis of the set of Microviridae phages defined around phi MH2K shows that phage classification based on adaptation does not reflect bacterial phylogeny. PMID- 18286221 TI - The superior group of vessels in the falciform ligament: anatomical and radiological correlation. AB - AIM: The purpose of this study was to clarify the anatomical detail of the superior group of vessels in the falciform ligament in terms of the relationship with the internal thoracic vessels, inferior phrenic vessels, and the intrahepatic portal vein. MATERIALS AND METHODS: (1) Anatomical study: we dissected eight adult human cadavers (seven normal and one cirrhotic liver) to determine the relationship between the superior group of vessels in the falciform ligament, the internal thoracic vessels, and the inferior phrenic vessels. (2) Clinical study: we determined the origin and destination of the superior group of veins demonstrated in 8 of 4,006 patients with chronic liver disease who underwent the contrast enhanced CT scans. RESULTS: (1) Anatomical study: the superior group of vessels anastomosed the right (n = 4), left (n = 2), and both (n = 2) internal thoracic vessels. They also anastomosed the left (n = 4), right (n = 1), and both (n = 2) inferior phrenic vessels. (2) Clinical study: the origin of the veins was identified as the left medial branch (n = 4), left lateral branch (n = 1), both the lateral and medial branches (n = 1), and the vein from the umbilical portion (n = 2) of the left portal vein. The drainage vein was identified as the left (n = 3), right (n = 2), and the both (n = 1) internal thoracic veins. CONCLUSION: We demonstrated the anastomoses between the superior group of vessels of the falciform ligament, the internal thoracic vessels, the inferior phrenic vessels, and the intrahepatic portal vein. These pre-existing anastomoses would develop as porto-systemic shunt in patients with portal hypertension. PMID- 18286222 TI - Multidetector-row CT angiographic imaging of the celiac trunk: anatomy and normal variants. AB - PURPOSE: To evaluate the ability of MDCT reformations in describing the celiac trunk vascular anatomy and variations. MATERIALS AND METHODS: A total of 555 MDCT angiographies of the abdominal aorta performed between January 2002 and July 2005 were retrospectively reviewed to assess the celiac trunk vascular anatomy and variations. All the patients with pathological condition likely to affect normal vascular anatomy as well as CT exams technically inadequate were excluded from our study. RESULTS: A total of 524 MDCT angiographies of abdominal aorta were included in our study. The classical configuration of the celiac trunk was detected in 72.1%. The hepato-splenic trunk was detected in 50.4% of cases; the hepato-gastro-splenic trunk was detected in 19.4% of cases; the gastro-splenic trunk was detected in 2.3% of cases. The hepato-spleno-gastric trunk associated with hepatic arteries variants were found in 15.4%. The hepato-splenic trunk, the hepato-gastric trunk, the hepato-splenic-mesenteric trunk, and the spleno-gastric trunk were found in 2.7, 5, 0.4, and 3.6%, respectively. In 0.6%, we found an absent celiac trunk. CONCLUSION: The knowledge of the type of anatomical variants and their subtypes is fundamental for a correct pre-operative vascular planning in surgical or radiological abdominal procedures. Multidetector-row CT (MDCT) provides high-quality 3D-reconstructed images and allows non-invasive assessment of normal anatomy and anatomic variants of celiac trunk. PMID- 18286223 TI - Di-myo-inositol phosphate and novel UDP-sugars accumulate in the extreme hyperthermophile Pyrolobus fumarii. AB - The archaeon Pyrolobus fumarii, one of the most extreme members of hyperthermophiles known thus far, is able to grow at temperatures up to 113 degrees C. Over a decade after the description of this organism our knowledge about the structures and strategies underlying its remarkable thermal resistance remains incipient. The accumulation of a restricted number of charged organic solutes is a common response to heat stress in hyperthermophilic organisms and accordingly their role in thermoprotection has been often postulated. In this work, the organic solute pool of P. fumarii was characterized using 1H, 13C, and 31P NMR. Di-myo-inositol phosphate was the major solute (0.21 micromol/mg protein), reinforcing the correlation between the occurrence of this solute and hyperthermophily; in addition, UDP-sugars (total concentration 0.11 micromol/mg protein) were present. The structures of the two major UDP-sugars were identified as UDP-alpha-GlcNAc3NAc and UDP-alpha-GlcNAc3NAc-(4<--1)-beta-GlcpNAc3NAc. Interestingly, the latter compound appears to be derived from the first one by addition of a 2,3-N-acetylglucoronic acid unit, suggesting that these UDP-sugars are intermediates of an N-linked glycosylation pathway. To our knowledge the UDP disaccharide has not been reported elsewhere. The physiological roles of these organic solutes are discussed. PMID- 18286224 TI - Isolation, gene detection and solvent tolerance of benzene, toluene and xylene degrading bacteria from nearshore surface water and Pacific Ocean sediment. AB - BTX (benzene, toluene and xylene) degrading bacteria were isolated from Pacific Ocean sediment and nearshore surface water. In the seawater near a ferry dock, degrading bacteria of a relatively wide diversity were detected, including species of Pseudomonas, Rhodococcus, Exiguobacterium and Bacillus; while species of Bacillus only have been detected from the deep-sea sediment. Most of the isolates showed degradation to more than one compound. Generally better growth was obtained with p-xylene and ethylbenzene than with the other two. All the bacteria could tolerate and grow with the compounds at 5-20% (v/v). Both benzene and toluene degradation related genes had been successfully PCR cloned from the isolates of nearshore water, the detected benzene dioxygenase gene was identical among all the species and close to its soil counterpart. However, they were not detected in all the isolates from deep sea. Results in this report suggested that BTX degrading bacteria widely spread in marine environments and they might be of potentials in biotreatment of BTEX in saline environments. PMID- 18286225 TI - Segregation distortion of wild-type alleles at the Machado-Joseph disease locus: a study in normal families from the Azores islands (Portugal). AB - Machado-Joseph disease (MJD) is caused by an expansion of a triplet repeat with a CAG motif at the ATXN3 gene. The putative segregation ratio distortion (SRD) of alleles can play an important role in the non-Mendelian behaviour of triplet repeat loci. To study the stability and infer the segregation patterns of wild type MJD alleles, the size of the (CAG)(n) tract was analysed in 102 normal sibships, representing 428 meioses. No mutational events were detected during the transmission of alleles. Segregation analysis showed that the smaller alleles were preferentially transmitted (56.9%). Considering maternal meioses alone, such preference was still detected (55.7%) but without statistical significance. A positive correlation was observed for the difference in length between the two alleles constituting the transmitters' genotype (D) and the frequency of transmission of the smaller alleles. The results suggest that small D values are not enough to modify the probability of allele transmission. When transmissions involving genotypes with D or=5% of the general population) may decrease the chance of developing PD, this effect was minor in the Cretan population. PMID- 18286227 TI - Importance of depletion interactions for structure and dynamics of ferrofluids. AB - The influence of attractive depletion forces on the structure and dynamics of ferrofluids is studied by computer simulations. In the presence of a magnetic field, we find that sufficiently strong depletion forces lead to an assembly of particle chains into columnar structures with hexagonal ordering inside the columns. In a planar shear flow, this ordering is destroyed, leading to strong shear thinning behavior. A pronounced anisotropy of the shear viscosity is observed. The viscosity is found to be largest when the magnetic field is oriented in the gradient direction of the flow. PMID- 18286228 TI - Static Rouse modes and related quantities: corrections to chain ideality in polymer melts. AB - Following the Flory ideality hypothesis intrachain and interchain excluded-volume interactions are supposed to compensate each other in dense polymer systems. Multichain effects should thus be neglected and polymer conformations may be understood from simple phantom chain models. Here we provide evidence against this phantom chain, mean-field picture. We analyze numerically and theoretically the static correlation function of the Rouse modes. Our numerical results are obtained from computer simulations of two coarse-grained polymer models for which the strength of the monomer repulsion can be varied, from full excluded volume ("hard monomers") to no excluded volume ("phantom chains"). For nonvanishing excluded volume we find the simulated correlation function of the Rouse modes to deviate markedly from the predictions of phantom chain models. This demonstrates that there are nonnegligible correlations along the chains in a melt. These correlations can be taken into account by perturbation theory. Our simulation results are in good agreement with these new theoretical predictions. PMID- 18286229 TI - Electrowetting --a versatile tool for controlling microdrop generation. AB - Integrating insulator-covered electrodes into a microfluidic flow focusing device (FFD) we demonstrate enhanced flexibility and control of the flow of two non miscible liquids based on electrowetting (EW). In the parameters space, determined by liquid inlet pressures, we identify a specific region where drops can only be generated and addressed via EW. In this regime we show that the size distribution and the frequency of drop generation can be controlled by the applied voltage and the width of voltage pulses. Moreover it turns out that with EW the drop size and the frequency can be tuned independently. Finally we show that the same drop generation phenomena can also be observed in the presence of surfactants. PMID- 18286230 TI - A model of gravity-induced distribution of material in plasma polymerized aerosols and films. AB - A mathematical model of the volumetric part of plasma polymerization influenced by gravity is presented. Plasma-activated adhesion of monomer molecules to a surface of a germinal particle is assumed as a basic mechanism of particulate growth. The continuity equation for the flow of matter through the discharge has been formulated and solved in two extreme asymptotic approximations--for small and major duration of the process. Several non-equilibrium distribution functions of the polymer were obtained, for instance, an amount of the particles as a function of their size or time of fall. Within the adopted model this function demonstrates a sharp downward increase inside a discharge. In addition it contains such parameters as the free fall acceleration or reaction rate coefficients, variations of which enable control of the discharge and properties of the disperse medium. PMID- 18286231 TI - Cinnarizine in refractory migraine prophylaxis: efficacy and tolerability. A comparison with sodium valproate. AB - This was a double-blind clinical trial designed to assess the efficacy and safety of the cinnarizine (CIN) in patients with migraine who were refractory to propranolol and tricyclic antidepressants in comparison with sodium valproate (SV) to investigate whether CIN could be at least as effective as SV. A total of 125 patients were treated in a treatment period of 12 weeks. All patients had at least one intake of trial medication and 2-week post baseline efficacy observation which all were included in the ITT analysis. Of the 125 subjects treated, 46 discontinued prematurely: 25 from the CIN and 21 from the SV group. The main reasons for premature discontinuation were: lost to follow up (25/46, 63.2%), insufficient response (16/46, 20%), and adverse events (5/46, 12.8%). No statistically significant inter-group differences in the number of discontinuation was observed (p > 0.05). In both groups, number of attacks, intensity, and duration of attacks significantly decreased (p < 0.05). No statistically significant inter-group differences were observed regarding the mean number of attacks, duration, and intensity of migraine attacks for any of the time intervals analysed, except for the mean reduction of third and fourth visits intensity from baseline which were significantly different in two groups (p < 0.05), with the CIN group showing more reduction. Analysis of the number of responders showed that in the CIN group 61.2% subjects were responders, and 63.8% in the SV group. No statistically significant differences between the treatment groups were found for any of the secondary parameters. Overall 26 subjects reported one or more adverse events during the study period: 13 subjects in each group. Five subjects discontinued prematurely due to adverse events; two in the CIN group with significant weight gain, and 3 in the SV group with significant weight gain and severe tremor. These results suggest that CIN is an effective and safe prophylactic agent even in severe migraine headache. PMID- 18286232 TI - Long term outcomes in patients with intracranial germinomas: a single institution experience of irradiation with or without chemotherapy. AB - Complete remission can be achieved soon after irradiation in patients with intracranial germinoma. This study aimed to analyze the follow-up outcome of intracranial germinoma patients. About 39 intracranial germinoma patients (29 males and 10 females; average age, 15 years; range, 7-27 years) treated at Kyoto University Hospital from 1978 to 2004 were included in the study group. Six patients had multifocal disease at initial diagnosis, and 10 had human chorionic gonadotropin (HCG)-producing tumors. Thirteen patients were treated with craniospinal axis irradiation, 6 with whole-brain irradiation, 17 with whole ventricle irradiation, and 3 with local field irradiation. Since 1997, 15 patients were treated with reduced-dose whole-ventricle irradiation (median, 23.4 Gy; range, 20.4-27 Gy) followed by a local boost (median, 40.8 Gy; range, 36-54 Gy) combined with chemotherapy. The median follow-up was 94 months (18 months to 25 years). The 5- and 10-year overall survival (OS) rates of the entire group were 97 and 90%, respectively. The 5- and 10-year progression-free survival (PFS) rates of the entire group were 91 and 87%, respectively. The 8-year OS and PFS in 15 patients treated by whole-ventricle irradiation combined with chemotherapy were 100% and 92%, respectively. Four patients had recurrences within a median period of 59.5 months (51-85 months). All relapses occurred outside the radiation fields. Tumor site, tumor size, HCG production, multifocal disease and radiation dose to the primary site or whole ventricle did not significantly affect PFS. All initial recurrences of intracranial germinoma occurred at the distant site out of the radiation field. Our data suggested that reduced doses to the whole ventricle, combined with chemotherapy, should be sufficiently effective in patients with intracranial germinoma. PMID- 18286233 TI - MT4-(MMP17) and MT6-MMP (MMP25), A unique set of membrane-anchored matrix metalloproteinases: properties and expression in cancer. AB - The process of cancer progression involves the action of multiple proteolytic systems, among which the family of matrix metalloproteinases (MMPs) play a pivotal role. The MMPs evolved to accomplish their proteolytic tasks in multiple cellular and tissue microenvironments including lipid rafts by incorporation and deletions of specific structural domains. The membrane type-MMPs (MT-MMPs) incorporated membrane anchoring domains that display these proteases at the cell surface, and thus they are optimal pericellular proteolytic machines. Two members of the MT-MMP subfamily, MMP-17 (MT4-MMP) and MMP-25 (MT6-MMP), are anchored to the plasma membrane via a glycosyl-phosphatidyl inositol (GPI) anchor, which confers these enzymes a unique set of regulatory and functional mechanisms that separates them from the rest of the MMP family. Discovered almost a decade ago, the body of work on GPI-MT-MMPs today is still surprisingly limited when compared to other MT-MMPs. However, new evidence shows that the GPI-MT-MMPs are highly expressed in human cancer, where they are associated with progression. Accumulating biochemical and functional evidence also highlights their distinct properties. In this review, we summarize the structural, biochemical, and biological properties of GPI-MT-MMPs and present an overview of their expression and role in cancer. We further discuss the potential implications of GPI anchoring for enzyme function. Finally, we comment on the new scientific challenges that lie ahead to better understand the function and role in cancer of these intriguing but yet unique MMPs. PMID- 18286234 TI - The tyrosine phosphatase Shp2 (PTPN11) in cancer. AB - Diverse cellular processes are regulated by tyrosyl phosphorylation, which is controlled by protein-tyrosine kinases (PTKs) and protein-tyrosine phosphatases (PTPs). De-regulated tyrosyl phosphorylation, evoked by gain-of-function mutations and/or over-expression of PTKs, contributes to the pathogenesis of many cancers and other human diseases. PTPs, because they oppose the action of PTKs, had been considered to be prime suspects for potential tumor suppressor genes. Surprisingly, few, if any, tumor suppressor PTPs have been identified. However, the Src homology-2 domain-containing phosphatase Shp2 (encoded by PTPN11) is a bona fide proto-oncogene. Germline mutations in PTPN11 cause Noonan and LEOPARD syndromes, whereas somatic PTPN11 mutations occur in several types of hematologic malignancies, most notably juvenile myelomonocytic leukemia and, more rarely, in solid tumors. Shp2 also is an essential component in several other oncogene signaling pathways. Elucidation of the events underlying Shp2-evoked transformation may provide new insights into oncogenic mechanisms and novel targets for anti-cancer therapy. PMID- 18286235 TI - Exopolysaccharides produced by Gordonia alkanivorans enhance bacterial degradation activity for diesel. AB - Exopolysaccharides (EPS) produced by Gordonia alkanivorans CC-JG39 was used to stimulate cell floating, cell growth, and diesel biodegradation of indigenous or commercial-available, diesel-degrading bacteria. Addition of EPS-containing supernatant into the culture medium resulted in floatation of the non-floating bacteria and allowed a 40-45% and 38-42% increase in diesel degradation and cell growth, respectively. The EPS-stimulating effect on cell growth and diesel degradation positively correlated with the EPS dosage. Thus, the EPS may act as a biostimulant for bioremediation of oil-contaminated water or soil. PMID- 18286236 TI - A comparative evaluation of three washing procedures for minimizing exogenous trace element contamination in fetal scalp hair of various obstetric outcomes. AB - A comparative evaluation of three washing methods for removing exogenous elemental contamination from fetal scalp hair is described. A total of 12 samples, including 6 from normal pregnancies and 6 from abruptio placentae (AP) cases, were used. Each was divided into four subsamples, with one left unwashed for comparison, while three were subjected to one of the three washing methods. The first consisted of agitating hair for 5 min in sequential and equal measures (30 ml) of acetone and water, followed by rinsing with acetone, and the process repeated twice. In the second and third, hair was agitated for 30 min in aqueous solutions of sodium lauryl sulfate (0.5%, w/v), and Triton X-100 (0.5%, v/v), respectively, followed by sequential rinsing with deionized water and acetone. After drying and microwave-assisted digestion in concentrated nitric acid, the subsamples' concentrations of zinc, copper, cadmium, lead, mercury and selenium were determined using inductively coupled mass spectrometry. All washed subsamples, irrespective of washing method used, had lower elemental concentrations than the corresponding, unwashed subsamples. These differences, taken as presumptive indicators of exogenous contamination, showed only small variations (<5%) between the three washing methods, irrespective of element, mode of delivery, and obstetric outcome. However, the contamination levels elicited by each of the three methods varied in magnitude according to individual elements, with those of copper being highest, followed by those of zinc, cadmium, mercury, selenium, and lead. They also varied according to obstetric outcome, being highest in the AP subsamples, while in the normal pregnancy group, copper and zinc contamination levels in the subsamples from neonates delivered by elective caesarean section were higher than in those from normal-delivery neonates. The data from this study suggest that the three washing methods used have comparable efficacies in removing exogenous elemental contamination from fetal hair, further inferring that it is probably a simple biological matrix to decontaminate. Possible contributory factors are discussed and contrasted with those affecting adult hair. PMID- 18286237 TI - Ceruloplasmin, copper, selenium, iron, zinc, and manganese levels in normal and sulfite oxidase deficient rat plasma: effects of sulfite exposure. AB - A noticeable effect of sulfite treatment was observed on the plasma ceruloplasmin ferroxidase activity of rats with normal sulfite oxidase activity when compared to normal controls. The plasma levels of selenium, iron, and zinc were unaffected by sulfite in normal and sulfite oxidase (SOX)-deficient rats. While plasma level of Mn was decreasing, plasma Cu level increased in SOX-deficient rats. Treating SOX-deficient groups with sulfite did not alter plasma level of Mn but made plasma level of Cu back to its normal level. This is the first evidence that Cu and Mn status were affected in experimental sulfite oxidase deficiency induced by low molybdenum diet with tungsten addition deserving further research to determine the underlying mechanisms of these observations in experimental sulfite oxidase deficiency. PMID- 18286238 TI - Trace element profiles in single strands of human hair determined by HR-ICP-MS. AB - Trace element analysis of human hair has the potential to reveal retrospective information about an individual's nutritional status and exposure. As trace elements are incorporated into the hair during the growth process, longitudinal segments of the hair may reflect the body burden during the growth period. We have evaluated the potential of human hair to indicate exposure or nutritional status over time by analysing trace element profiles in single strands of human hair. The hair strands from five healthy and occupationally unexposed subjects were cut into 1-cm long segments starting from the scalp. By using high resolution inductively coupled plasma mass spectrometry (HR-ICP-MS), we achieved profiles of 12 elements in single strands of human hair, namely, Ag, As, Au, Cd, Cu, Hg, Fe, Pb, Se, Sr, U and Zn. We have shown that trace element analysis along single strands of human hair can yield information about essential and toxic elements, and for some elements, can be correlated with seasonal changes in diet and exposure. The information obtained from the trace element profiles of human hair in this study substantiates the potential of hair as a biomarker. PMID- 18286239 TI - Blood lead levels and increased bronchial responsiveness. AB - The immune system is one of the targets most sensitive to lead toxicity, and the association between lead exposure and serum immunoglobulin E (IgE) has been published. Recent studies also reported that lead caused the development of IgE mediated allergy. To investigate whether blood lead levels contribute to other allergic conditions, we examined the effect of blood lead on bronchial responsiveness (BR) in the general population. We performed a cross-sectional study with adults aged 19 to 58 years in a Korean community. Blood lead level and the methacholine provocation test were performed. The overall mean blood lead level was 2.9 microg/dl, and the mean BR index was 1.14. The percent of subjects with clinically diagnosed asthma was 21 (4%) and there was no difference in blood lead level with and without asthma. In the multiple regression model, the elevation of blood lead level was related to the increase of BR after adjusting for age, sex, height, smoking status, and the presence of asthma. Blood lead level was significantly associated with increased BR that came from the elevation of the IgE level with lead exposure. Lead may contribute to the increase of asthma and other allergic conditions. PMID- 18286240 TI - The protein-binding potential of C2H2 zinc finger domains. AB - There are over 10,000 C2H2-type zinc finger (ZF) domains distributed among more than 1,000 ZF proteins in the human genome. These domains are frequently observed to be involved in sequence-specific DNA binding, and uncharacterized domains are typically assumed to facilitate DNA interactions. However, some ZFs also facilitate binding to proteins or RNA. Over 100 Cys2-His2 (C2H2) ZF-protein interactions have been described. We initially attempted a bioinformatics analysis to identify sequence features that would predict a DNA- or protein binding function. These efforts were complicated by several issues, including uncertainties about the full functional capabilities of the ZFs. We therefore applied an unbiased approach to directly examine the potential for ZFs to facilitate DNA or protein interactions. The human OLF-1/EBF associated zinc finger (OAZ) protein was used as a model. The human O/E-1-associated zinc finger protein (hOAZ) contains 30 ZFs in 6 clusters, some of which have been previously indicated in DNA or protein interactions. DNA binding was assessed using a target site selection (CAST) assay, and protein binding was assessed using a yeast two hybrid assay. We observed that clusters known to bind DNA could facilitate specific protein interactions, but clusters known to bind protein did not facilitate specific DNA interactions. Our primary conclusion is that DNA binding is a more restricted function of ZFs, and that their potential for mediating protein interactions is likely greater. These results suggest that the role of C2H2 ZF domains in protein interactions has probably been underestimated. The implication of these findings for the prediction of ZF function is discussed. PMID- 18286244 TI - Genetics of autoimmune diabetes mellitus. AB - Autoimmune diabetes mellitus, called type 1 diabetes mellitus (T1DM), is caused by autoimmune destruction of islet beta cells in the pancreas. T1DM susceptibility loci mapped by different genome screening are IDDM1-IDDM18. It has been estimated that HLA (IDDM1) provides up to 40-50 % of the familial clustering of T1DM (LOD score of 65.8). Many studies have verified that DQB1*0302 is a strong susceptibility gene and that the heterozygous combination of DQA1*0301 DQB1*0302 on the HLA-DR4 haplotype and DQA1*0501-DQB1*0201 on the HLA-DR3 haplotype results in a synergistically increased risk of T1DM. The presence of predisposing genes in autoimmune diabetes decreases with age, probably due to increasing influence of environmental factors. Autoimmune diabetes with manifestation in adults may have partly different immunogenetic etiopathogenesis than autoimmune diabetes with manifestation in childhood. Compared to fast progressing adult-onset T1DM, slowly progressing adult-onset type 1 diabetes (LADA) might involve genes leading to a slow progressive beta-cells destruction. PMID- 18286245 TI - [From the allergen-recognition by antibodies to new therapeutic concepts]. AB - Cross-linking of IgE antibodies through allergens is a basic event in type I allergy and leads to the immediate release of mediators like histamine, responsible for allergic symptoms like rhino-conjunctivitis or asthma. Critical for the binding of allergens to IgE are the IgE-epitopes, which represent a congregation of several amino acid residues often derived from different regions of the allergen. By means of the mimotope-technology, we isolated peptides from phage libraries, which were able to structurally mimic IgE-epitopes of the plant allergens Bet v 1 (birch) and Phl p 5a (timothy grass). Hence, these are candidates for an epitope-specific immunotherapy. In this mode of immunotherapy, it is the aim to induce IgG antibodies directed exclusively against the IgE epitopes of allergens without induction of anaphylactogenic IgG species, and without the risk of anaphylaxis through cross-linking of IgE. Immunizing mice, we applied the mimotopes displayed on bacteriophages as well as on alternative carrier systems to enhance their antigenicity. With these systems it was possible to elicit an allergen-specific immune response, which was also accompanied by the appropriate T-cell help. Mimotopes resemble a promising concept for an epitope tailored immunotherapy of allergic patients. PMID- 18286246 TI - Autoantibodies in systemic sclerosis (scleroderma): clues for clinical evaluation, prognosis and pathogenesis. AB - Systemic sclerosis is a generalized autoimmune connective tissue disease of unknown aetiology. Profound vascular and immunological dysregulations result in tissue fibrosis affecting the skin and internal organs. Currently, two main clinical subtypes are distinguished, i.e. limited and diffuse cutaneous systemic sclerosis, which differ significantly in the clinical course and prognosis. Autoantibodies against topoisomerase (Scl-70), centromere-associated proteins, and nucleolar antigens are important for the diagnosis of the disease and give clues for its clinical manifestations and prognosis (prognostic autoantibodies). For most of these antibodies, however, the role in pathogenesis is not established. Anti-centromere antibodies are associated with limited cutaneous involvement and risk for pulmonary hypertension, whereas anti-topoisomerase I is associated with diffuse progressive disease and severe interstitial lung disease. Anti-Th/To positivity is associated with limited skin involvement but a high risk for severe internal organ involvement (Kidneys, PAH, Lung fibrosis). Anti-RNA polymerase I/III antibodies are associated with a high risk for renal involvement. Autoantibodies against the PDGF receptor and fibrillin-1 seem to play important roles in the pathogenetic process of systemic sclerosis. PMID- 18286247 TI - [Thyroid and treatment with amiodarone diagnosis, therapy and clinical management]. AB - Amiodarone is a frequently used antiarrhythmic drug with a high antiarrhythmic potency. However, beside its antiarrhythmic effects Amiodarone also reveals a variety of adverse effects and drug-related complications. The affected organs include the eyes, skin, lungs, nervous system, liver, gastrointestinal tract and the thyroid. The thyroid is one of the most frequently affected organs by Amiodarone. An altered hormone equilibrium always occurs and has to be distinguished from Amiodarone induced hyperthyroidism and hypothyroidism. The differentiation of these states frequently causes problems and may even be a diagnostic and therapeutic challenge in certain cases. The article gives an overview on the interactions between Amiodarone and the thyroid, the diagnostic and therapeutic options and management strategies of patient on Amiodarone therapy in the view of thyroid function. PMID- 18286248 TI - Mobile phones: are children at higher risk? AB - The use of mobile phones by children has in recent years been increasing, the onset starting earlier and earlier. This has led to concern about whether children might be at higher risk and whether mobile phone use should even be forbidden. Advice from highly reputed committees on both national and international levels is contradictory and on the whole more confusing than helpful to worried parents and physicians, but this can be explained. It is shown here that different opinions derived from the same database reflect the different way in which uncertainty of knowledge is dealt with. Based on the assessments of the international committee and established knowledge on children's development it can be concluded that existing exposure limits do in fact provide reasonable safety. Compared to adults, however, children do in fact need to be considered at a relatively higher potential risk, depending on their phase of development. If adults are advised to minimise their exposure, this is even more justified for children. However, circular reasoning must be avoided: the uncertainty-triggered application of precaution must not be misinterpreted as evidence for definite risk. PMID- 18286249 TI - Should hospitals protect from geopathogenic zones? AB - The belief that geopathogenic zones exist and could cause morbidity is widespread. It is increasing even among physicians. Presently, in health care it is going to even gain an economical dimension by causing investment costs for claimed protective means and by influencing hospital planning, design and management. Therefore, this issue is no longer just a private borderline issue but has become publicly relevant. In view of the fact that relevant studies are lacking, it deserves scientific clarification. In a double-blind study this hypothesis was tested by two different approaches: First, it was investigated whether location-bound biologic effects could be identified at all. To remain independent from dowsers, an investigation area of 35 m(2) was chosen which was large enough to ensure the presence of at least one postulated adverse zone. This area was scanned in 0.5 m intervals by monitoring bioparameters such as skin resistance, reaction time and heart rate variability in 43 volunteers. Skeptics were excluded. Location-dependent changes were investigated by point-by point analysis of local data distributions and by analysing the two-dimensional spatial distributions of the measured bioparameters by cross-correlation analysis. In a second approach, 5 independent experienced dowsers were asked to locate geopathogenic zones within the area both to verify the adequate choice of the area and to correlate the pattern of dowser's scorings with the patterns of biosignal distributions. Neither of these approaches showed statistical significant differences. The investigation did not confirm the hypothesis that local geopathogenic zones existed and, hence, would need protective actions. PMID- 18286250 TI - [First warnings of the dangers involved in tobacco use. Medical-scientific knowledge and recommendations for prevention in the second half of the 19th century and the first half of the 20th century]. AB - References to historical aspects of tobacco research and the ensuing recommendations for the prevention of tobacco-associated diseases can only be found rudimentarily in newer studies. This article systematically reviews the historical literature on the subject, with special attention given to selected journals published between 1850 and 1950. The analysis offers a chronological summary of the knowledge about tobacco ingredients and their effects, about tobacco-attributed diseases as well as of the usually non-institutionalized recommendations for their prevention. The report clearly shows the early knowledge about the toxic effects of tobacco and its products, the causation of ophthalmologic diseases and the effects on the cardiovascular system, the respiratory and nervous systems as well as the reproduction system. The knowledge of the pathogenesis of tobacco-attributable mental disorders, addiction and cancer is also reported. From today's view, some former strategies of prevention are regarded as obsolete; some seem to be very topical. In summary, research concentrating on the characteristics of tobacco and tobacco-associated diseases is witness to a continuity spanning different epochs and political systems. PMID- 18286251 TI - [Systematic literature search]. AB - Systematic literature searches in bibliographic databases are an essential step in constructing systematic reviews and health technology assessments. The purpose of this kind of search is to identify as many relevant references on a given topic in electronic databases as possible. This article begins with describing the search process and continues with the basics of searching electronic databases. By delivering insight into the functionalities of databases and possibilities to search them (particularly PubMed), an understanding of potentials and weaknesses of systematic literature searches should be established. PMID- 18286252 TI - [Anesthesia for carotid artery surgery. Is there a gold standard?]. AB - Carotid endarterectomy (CEA) has remained the first choice of treatment in preventing ischemic stroke due to symptomatic stenosis of the internal carotid artery despite other new available options. During CEA patients are first and foremost at risk of cerebral as well as myocardial ischemia, therefore, maintenance of the oxygen supply to the brain and the myocardium is of outstanding importance requiring reliable monitoring of cerebral and cardiac function. The regional versus general anesthesia debate is an age-old one that has brought few definite answers regarding the impact on postoperative outcome of either anesthetic technique. Up to now, there is little evidence from only a few randomized clinical trials to demonstrate the superiority of either anesthetic technique with respect to outcome parameters. Because the level of evidence in favor of regional anesthesia during CEA can at least be rated between 1(-) and 2(+) the resulting recommendation is grade C. The purpose of the review is to highlight the characteristics and goals of anesthetic management during CEA. PMID- 18286254 TI - [Proliferation and differentiation of human osteoblasts from the nasal septum in a new perfusion culture system]. AB - BACKGROUND: Today, perfusion culture systems are mainly used to investigate cellular physiology and to cultivate three-dimensional tissue complexes. As a rule, these systems are relatively expensive and do not enable continuous microscopic monitoring of the growing cells. Simple and inexpensive perfusion culture systems have not been available up to now. METHODS: A novel perfusion culture system was developed in which the modular components consist of a mounting apparatus for inserting various media supply systems, microdispenser pumps, and laminar-flow culture chambers, each with a culture volume of 8 cm(3). The perfusion chambers were inoculated with human osteoblast cells from the tissue culture (5,000/cm(2)) and were perfused for 10 days after adherence of the cells (0.5 ml/min). As a control group, osteoblast-like cells were cultured in identically constructed culture chambers without medium perfusion. After 10 days, the cell counts were determined in accordance with the Coulter principle. Alkaline phosphatase was measured photometrically as a characteristic for differentiation. RESULTS: Compared with the control group, three to four times the quantity of cells were produced within 10 days in the perfusion cultures. The alkaline phosphatase values were equally high or only slightly lower, indicating that osteoblast differentiation of the cells was maintained with a higher proliferation. CONCLUSIONS: As large a number of in vitro proliferated cells as possible is a prerequisite for clinical application of tissue engineering. By continuously supplying medium, the tested perfusion culture system enables a higher rate of proliferation of osteoblast-like cells with maintenance of differentiation. Continuous microscopic monitoring of the cultures is possible using commercially available Petri dishes. PMID- 18286253 TI - [Tissue engineering of respiratory epithelium. Regenerative medicine for reconstructive surgery of the upper airways]. AB - Reconstruction of long tracheal defects remains an unsolved surgical problem. Tissue engineering of respiratory epithelium is therefore of utmost surgical and scientific interest. Successful cultivation and reproduction of respiratory epithelium in vitro is crucial to seed scaffolds of various biomaterials with functionally active respiratory mucosa. Most frequently, the suspension culture as well as the tissue or explant cultures are used. Collagenous matrices, synthetic and biodegradable polymers, serve as carriers in studies. It is essential for clinical practice that mechanically stable biomaterials be developed that are resorbable in the long term or that cartilaginous constructs produced in vitro be employed which are seeded with respiratory epithelium before implantation. Vascularization of a bioartificial matrix for tracheal substitution is also prerequisite for integration of the constructs produced in vitro into the recipient organism. Here, the state of the art of research, perspectives and limitations of tracheal tissue engineering are reviewed. PMID- 18286255 TI - [Brachytherapy of the prostate cancer]. AB - Prostate cancer (PCA) is the most frequent onlocological disease in men. Every year there are ca. 202.000 new cases of prostate cancer in Europe. Curative treatment of this carcinoma via brachytherapy is becoming increasingly significant (20-30% of all curative approaches). Initial staging and thus allocation to risk groups prior to the commencement of therapy is esspecially important for successful brachytherapy treatment.Low-dose-rate (LDR) brachytherapy (i.e. SEED implantation) distinguishes itself both with respect to the procedure as well as the indication from high-dose-rate brachytherapy (afterloading procedure). Both treatment procedures are employed as monotherapy as well as in combination with external radiation.LDR monotherapy is reported to achieve biochemically relapse-free outcome of up to 90% in low-risk tumours during 10-year follow-up periods. Combined HDR tele- and brachytherapy is reported to achieve a biochemically relapse-free outcome of 80-90% with intermediate- and high-risk tumours in long-term follow-up.While randomized studies are as yet missing, it is still possible to derive the following application algorithms from monitoring studies and cohort studies: application of LDR monobrachytherapy must be restricted to low-risk tumorus. Combined HDR tele- and brachytherapy can be sucessfully applied in cases of intermediate- and high risk tumours. The outcome depends significantly on the initial, pre-therapy PSA value and Gleason score. Posttherapeutically, the nadir value is crucial with respect to predicting the biochemically relapse-free outcome. PMID- 18286256 TI - Self-monitoring of blood glucose in type 2 diabetes: a new look at published trials. PMID- 18286257 TI - Generalised reduction of putative endothelial progenitors and CXCR4-positive peripheral blood cells in type 2 diabetes. AB - AIMS/HYPOTHESIS: In patients with type 2 diabetes, reduced levels of circulating endothelial progenitor cells have been reported and these have been correlated with disease severity. In this study, we examined a panel of markers widely used to identify progenitor and/or stem cells, and determined their association with disease severity in diabetic patients. Since expression of chemokine (C-X-C motif) receptor 4 (CXCR4) has been associated with mobilisation and recruitment of progenitor cells, CXCR4 expression was also analysed. METHODS: Peripheral blood mononuclear cells (PBMCs) from 98 patients with type 2 diabetes and 39 control individuals were analysed by flow cytometry for surface marker expression. RESULTS: Cells expressing different combinations of progenitor and/or stem cell markers were severely reduced in PBMCs of diabetic patients compared with those of control participants. Moreover, a number of these putative progenitor cell populations were negatively associated with disease severity. Reduced expression of CXCR4 and CD34/CXCR4-positive cells was also observed in diabetic patients. PBMCs expressing CXCR4 positively correlated with levels of progenitor cells in control participants but not in diabetic patients. Levels of putative progenitor and CXCR4-positive cells were further decreased in patients with diabetic complications, including cardiovascular and microvascular diseases. CONCLUSIONS/INTERPRETATION: A generalised decrease in a range of progenitor cell populations was observed in type 2 diabetic patients. This reduction was also negatively associated with disease severity. PMID- 18286259 TI - [Introduction to the topic. Cartilage damage of the ankle joint. New and well established approaches to treatment]. PMID- 18286258 TI - Low hepatic stearoyl-CoA desaturase 1 activity is associated with fatty liver and insulin resistance in obese humans. AB - AIMS/HYPOTHESIS: Stearoyl-CoA desaturase 1 (SCD1) is the rate-limiting enzyme in monounsaturated fatty acid synthesis. It is imperative for the assembly of VLDL particles, which transport triacylglycerol (TG) from liver to adipose tissue and other sites. We aimed to determine the role of hepatic SCD1 activity in human glucose and lipid metabolism. METHODS: We studied 54 people participating in a lifestyle intervention programme with diet modification and increased physical activity. Insulin sensitivity was determined during a euglycaemic hyperinsulinaemic clamp and estimated from an OGTT. Liver fat was quantified by (1)H-magnetic resonance spectroscopy at baseline and after 9 months of intervention. The pattern of fatty acids in serum VLDL-TGs was determined by ultracentrifugation followed by thin layer and gas chromatography, with the 18:1 n-9: 18:0 ratio providing an index of hepatic SCD1 activity. RESULTS: The hepatic SCD1 activity index correlated negatively with liver fat (r= -0.29, p=0.04) and positively with insulin sensitivity, both OGTT-derived (r=0.42, p=0.003) and clamp-derived (r=0.27, p=0.07). These correlations depended on overall adiposity. They were absent in leaner participants (n=27, liver fat: p=0.34, insulin sensitivity [OGTT]: p=0.75, insulin sensitivity [clamp]: p=0.24), but were strong in obese individuals (n=27, p=0.004, p=0.0002 and p=0.006, respectively). Furthermore, during intervention a high SCD1 activity index at baseline predicted a decrease in liver fat only in obese participants (r= -0.46, p=0.02). CONCLUSIONS/INTERPRETATION: Our data suggest that high hepatic SCD1 activity may regulate fat accumulation in the liver and possibly protects from insulin resistance in obesity. PMID- 18286260 TI - [Osteotomies of the distal tibia and hindfoot for ankle realignment]. AB - Asymmetric load of the ankle joint often results in degenerative disease. Although reconstructive surgery, including osteotomies above and beneath the tibiotalar joint, is possible, there are little data with respect to its evidence. This article presents general considerations for osteotomies around the osteoarthritic ankle and elaborates a rationale for the technical procedure. Additional measures for correcting the deformed and malaligned hindfoot are also elucidated. As a principle, opening-wedge and closing-wedge osteotomies are possible in one or more planes. In some instances, inframalleolar osteotomies are also necessary to achieve proper alignment of the foot. If present, imbalance of soft tissues, such as incompetence of ligaments and insufficiency of tendons, must also be addressed. Our results have shown that osteotomies above and beneath the ankle joint are able to correct deformities and incongruencies at the tibiotalar joint over the years, thus avoiding further cartilage wear. In some patients, the tibiotalar joint regained a regular joint space that can be attributed to potential regeneration of cartilage. In all but a few cases (<5%), arthrodesis or total ankle replacement has been successfully avoided. This benefit is even more important because mostly younger, active patients are involved, and long-term results after arthrodesis and total ankle replacement are critical. Therefore, our treatment strategy is to correct the deformity first to achieve a well-aligned and balanced tibiotalar joint. If necessary, total ankle replacement is considered in a second stage. PMID- 18286261 TI - [Autologous chondrocyte transplantation in the ankle joint. Rational or irrational?]. AB - Ankle sprains are one the most common injuries of the lower limb. Fractures, ligamentous lesions, and cartilaginous damage are often associated. Nevertheless the injury is often misjudged and concomitant chondral lesions are assessed late. In the case of a symptomatic osteocartilaginous lesion of the talus, which can be illustrated by MRI or X-ray, operative intervention is indicated. Methods such as microfracturing, mosaicplasty, and autologous chondrocyte transplantation (ACT) are in clinical use. The latter is well known and being established as the treatment of choice for large cartilage defects in the knee. Due to the good results in the knee and the technological improvements (three-dimensional tissue constructs seeded with autologous chondrocytes) this method is being increasingly applied for cartilage lesions of the talus. In contrast to the mosaicplasty donor site morbidity is low and the size of the defect is not a limiting factor. The current studies about ACT of the talus show a stable repair of the defect with mostly hyaline-like cartilage and high patient satisfaction. Therefore, the procedure can be recommended for lesions>1 cm2. Concomitant treatment of posttraumatic deformities (malalignment), ligamentous instabilities, and especially the reconstruction of bony defects are compulsory. PMID- 18286263 TI - The new Surviving Sepsis Campaign recommendations on glucose control should be reconsidered. PMID- 18286264 TI - Haem oxygenase: a model for therapeutic intervention. PMID- 18286265 TI - Use of carbon monoxide as a therapeutic agent: promises and challenges. AB - As a by-product of heme catabolism by the heme oxygenase system, carbon monoxide (CO) has been neglected for many years, and only recently has its role as an essential signaling molecule been appreciated. In the past decade, the use of CO gas in pre-clinical experimental models of disease has produced some remarkable data indicating that its therapeutic delivery to mammals could alleviate inflammatory processes and cardiovascular disorders. However, the inherent toxic nature of CO cannot be ignored, knowing that inhalation of uncontrolled amounts of this gas can ultimately lead to serious systemic complications and neuronal derangements. From a clinical perspective, a key question is whether a safe and therapeutically effective threshold of CO can be reached locally in organs and tissues without delivering potentially toxic amounts through the lung. The advent of CO-releasing molecules (CO-RMs), a group of compounds capable of carrying and liberating controlled quantities of CO in cellular systems, appears a plausible alternative in the attempt to overcome the limitations of CO gas. Although in its infancy and far from being used for clinical applications, the CO-RMs technology is supported by very encouraging biological results and reflected by the chemical versatility of these compounds and their endless potential to be transformed into CO-based pharmaceuticals. PMID- 18286267 TI - Addition of exogenous carbon and nitrogen sources to aphid exuviae modulates synthesis of proteases and chitinase by germinating conidia of Beauveria bassiana. AB - Secretion of catabolic extracellular enzymes (ECE) is the hallmark of the infection of insects through the cuticle by entomopathogenic fungi (EPF). In this paper, we show that germinating conidia of Beauveria bassiana (Bb) regulate the synthesis of ECE through a multiple control mode during the initial stages of germination. We tested Bb conidial growth on aphid exuviae with or without supplementation of additional carbon and/or nitrogen (C/N) compounds. To understand the interrelation between conidial germination during growth, the synthesis of ECE activity, free amino nitrogen (FAN), glucose and fungal dry weight biomass were measured. Immediately (0.25 h) upon incubation of conidia, activity of subtilisin-like Pr1 and trypsin-like Pr2 enzymes and chitinase (NAGase) was observed in the culture filtrates. At 0.25 h, addition of exogenous C-source resulted in higher activities of Pr1 and Pr2, respectively. Conversely at 0.25 h, addition of N-sources repressed the synthesis of Pr2, but that of Pr1. C/N repression was observed only for exponentially growing mycelia. NAGase activity remained at basal level and unaffected by added C/N. We conclude that C/N repression occurs only when it is necessary for the Bb infective structures to establish a nutritional relationship with the host structures. PMID- 18286266 TI - The heme oxygenase-carbon monoxide system: regulation and role in stress response and organ failure. AB - Heme oxygenase (HO) breaks down heme, the iron-containing, oxygen-carrying constituent of red blood cells, yielding biliverdin, iron (II) ions, and carbon monoxide (CO). Among the isoenzymes cloned to date, only HO-1 can be induced by a panoply of stimuli linked by their ability to provoke oxidative stress. HO-1 induction protects against cell death in experimental models associated with ischemia/reperfusion or inflammation, making the gene a promising target for critical care medicine. Induction of HO-1 may confer protection by controlling intracellular levels of toxic heme, or by anti-inflammatory, anti-apoptotic, and blood flow-maintaining effects of its by-products biliverdin and CO. Although protective effects of upregulation of HO-1 have been reported for a variety of cells and tissues, evidence suggests that the protective action may be restricted to a rather narrow threshold of overexpression. In addition, there is substantial variation in gene expression depending on transcriptional control mechanisms such as a microsatellite length polymorphism. Genetic variability and the required use of cytotoxic inducers are hurdles for purposeful targeting of HO-1 gene expression in critical care, while administration of by-products of the pathway seems feasible at present. PMID- 18286268 TI - Screening of dissolved heavy metals (Cu, Zn, Mn, Al, Cd, Ni, Pb) in seawater by a liquid-membrane-ICP-MS approach. AB - A bulk liquid membrane system has been developed and applied to the simultaneous separation and preconcentration of up to seven heavy metals (copper, zinc, lead, cadmium, aluminium, manganese, and nickel) in seawater. Copper was selected to optimize transport conditions and then, under these conditions, the simultaneous extraction of other heavy metals was studied. The system achieved preconcentration yields ranging between 44.11% (Cd) and 77.77% (Cu) after nine hours of operation, the effectiveness of metal transport being Cu > Zn > Pb > Mn > Ni > Al > Cd. The system was applied to the preconcentration of four real seawater samples before their quantification by inductively coupled plasma-mass spectrometry (ICP-MS). Compared with the analytical procedures commonly used for trace metal determination in oceanography, the results obtained demonstrated that the new system may be used as a very clean (sample contamination-free), simple, and one-step alternative for semiquantitative, and even quantitative, simultaneous determination of heavy metals in seawater. PMID- 18286269 TI - Label-free detection with micro optical fluidic systems (MOFS): a review. AB - The paper reviews the state-of-art for micro optical fluidic systems (MOFS), or optofluidics, which employs optics and fluidics in a microsystem environment to perform novel functionalities and in-depth analysis in the biophysical area. Various topics, which include the introduction of MOFS in biomedical engineering, the implementation of near-field optics and also the applications of MOFS to biophysical studies, are discussed. Different optical detection techniques, such as evanescent wave, surface plasmon resonance, surface enhanced Raman scattering, resonators and transistors, have been studied extensively and integrated into MOFS. In addition, MOFS also provides a platform for various studies of cell biophysics, such as cell mass determination and cell Young's modulus measurement. PMID- 18286270 TI - The future of stone research: rummagings in the attic, Randall's plaque, nanobacteria, and lessons from phylogeny. AB - The prevention or cure of stone disease will be achieved only by identifying biochemical, physiological and molecular mechanisms operating before the formation of a calculus. Yet, the gradual increase in the total number of papers devoted to the study of kidney stones that has occurred since the beginning of the 21st century can be attributed almost entirely to papers concerned with the investigation of factors associated with urolithiasis after stones have already formed. The need to prevent stones by discovering how the human body routinely stops their formation in those of us who do not suffer from them is therefore as exigent as ever and a new approach to investigating the causes of stones is urgently needed. In this paper, I develop the view that stone research will best progress by examining and understanding how healthy plants and animals control the formation of biominerals. In addition to structures like bones, teeth, shells and spines, many organisms spanning the entire phylogenetic tree form intra- and extracellular granules which are use as storage depots for calcium and other important ions, which they can reclaim to maintain homeostasis or to satisfy specific needs during periods of high demand, such as shell formation, moulting or skeletal development. These electron-dense granules, which also bear an uncanny resemblance to calcified nanobacteria, are remarkably similar in general structure, size and composition to particles observed in healthy human kidneys and in Randall's plaque. Therefore, it is likely that the granules in human kidneys fulfil analogous functions to those in other organisms-particularly in calcium homeostasis. Their study in a large range of creatures has already provided a deep well of information about their structure, movement, composition, macromolecular content, synthesis and resorption, from which we can draw to quench our thirst for knowledge of basic mechanisms and events involved in the formation of human kidney stones. PMID- 18286271 TI - Multiple short-segment colonic duplications. AB - Gastrointestinal tract duplication is an uncommon congenital anomaly that rarely involves the colon. We report an unusual case of multiple short-segment duplications of the colon not previously reported in the literature. The patient had a large diverticulum-type duplication of the descending and sigmoid colon and two other tubular duplications of the transverse and ascending colon, the latter two communicating with the colon at both ends of the duplications. The symptomatic diverticulum-type of duplication was surgically treated. PMID- 18286272 TI - Singlet oxygen generation in the reaction centers of Rhodobacter sphaeroides. AB - Singlet oxygen (1O2) generation in the reaction centers (RCs) of Rhodobacter sphaeroides wild type was characterized by luminescent emission in the near infrared region (time resolved transients and emission spectra) and quantified to have quantum yield of 0.03 +/- 0.005. 1O2 emission was measured as a function of temperature, ascorbate, urea and potassium ferricyanide concentrations and as a function of incubation time in H2O:D2O mixtures. 1O2 was shown to be affected by the RC dynamics and to originate from the reaction of molecular oxygen with two sources of triplets: photoactive dimer formed by singlet-triplet mixing and bacteriopheophytin formed by direct photoexcitation and intersystem crossing. PMID- 18286273 TI - Dynamics of hydration water in deuterated purple membranes explored by neutron scattering. AB - The function and dynamics of proteins depend on their direct environment, and much evidence has pointed to a strong coupling between water and protein motions. Recently however, neutron scattering measurements on deuterated and natural abundance purple membrane (PM), hydrated in H2O and D2O, respectively, revealed that membrane and water motions on the ns-ps time scale are not directly coupled below 260 K (Wood et al. in Proc Natl Acad Sci USA 104:18049-18054, 2007). In the initial study, samples with a high level of hydration were measured. Here, we have measured the dynamics of PM and water separately, at a low-hydration level corresponding to the first layer of hydration water only. As in the case of the higher hydration samples previously studied, the dynamics of PM and water display different temperature dependencies, with a transition in the hydration water at 200 K not triggering a transition in the membrane at the same temperature. Furthermore, neutron diffraction experiments were carried out to monitor the lamellar spacing of a flash-cooled deuterated PM stack hydrated in H2O as a function of temperature. At 200 K, a sudden decrease in lamellar spacing indicated the onset of long-range translational water diffusion in the second hydration layer as has already been observed on flash-cooled natural-abundance PM stacks hydrated in D2O (Weik et al. in J Mol Biol 275:632-634, 2005), excluding thus a notable isotope effect. Our results reinforce the notion that membrane protein dynamics may be less strongly coupled to hydration water motions than the dynamics of soluble proteins. PMID- 18286274 TI - Differences between the channels, currents and mechanisms of conduction slowing/block and accommodative processes in simulated cases of focal demyelinating neuropathies. AB - To clarify the differences between the mechanisms of conduction slowing/block and accommodative processes in focal demyelinating neuropathies, this computational study presents the kinetics of the ionic, transaxonal and transmyelin currents defining the intracellular and electrotonic potentials in different segments of human motor nerve fibres. The computations use our previous double cable model of the fibres. The simulated fibres have focal demyelination of internodes, paranodes or both together. The intracellular potentials are defined mainly by the Na+ current, as the contribution of the K+ fast and K+ slow currents to the total nodal ionic current is negligible. The paranodal demyelinations cause an increase in the transaxonal current and a decrease in the transmyelin current at the paranodal segments. However, there is an inverse relationship between the transaxonal and transmyelin currents at the same segments in the cases of internodal demyelination. The internodal ionic channels beneath the myelin sheath do not contribute to the intracellular potentials, but they show a high sensitivity to long-lasting pulses. The slow components of the electrotonic potentials depend on the activation of the channel types in the nodal or internodal axolemma, whereas the fast components of the potentials are determined mainly by the passive cable responses. However, the current kinetics changes (defining the investigated electrotonic changes) are relatively weak. The study summarizes the results from these modelling investigations on the mechanisms underlying the conduction slowing/block and accommodative processes in focal demyelinating neuropathies such as Guillain-Barre syndrome and multifocal motor neuropathy. PMID- 18286275 TI - An alternative theoretical formula for hemoglobin oxygenation. AB - Classical models of homotropic allostery are based on the postulate that the binding sites are equivalent in their ability to interconvert between high and low affinity states, but compelling evidence exists that the subunits of human hemoglobin are not simultaneously available for oxygen equilibration, thus reducing the number of possible intermediate microstates. The incorporation of these results into the Adair scheme reveals an alternative mechanism for hemoglobin oxygenation, not based on affinity changes. PMID- 18286276 TI - Solvent isotope effect on macromolecular dynamics in E. coli. AB - Elastic incoherent neutron scattering was used to explore solvent isotope effects on average macromolecular dynamics in vivo. Measurements were performed on living E. coli bacteria containing H2O and D2O, respectively, close to physiological conditions of temperature. Global macromolecular flexibility, expressed as mean square fluctuation (MSF) values, and structural resilience in a free energy potential, expressed as a mean effective force constant, [Symbol: see text]k'[Symbol: see text], were extracted in the two solvent conditions. They referred to the average contribution of all macromolecules inside the cell, mostly dominated by the internal motions of the protein fraction. Flexibility and resilience were both found to be smaller in D2O than in H2O. A difference was expected because the driving forces behind macromolecular stabilization and dynamics are different in H2O and D2O. In D2O, the hydrophobic effect is known to be stronger than in H2O: it favours the burial of non-polar surfaces as well as their van der Waals' packing in the macromolecule cores. This may lead to the observed smaller MSF values. In contrast, in H2O, macromolecules would present more water-exposed surfaces, which would give rise to larger MSF values, in particular at the macromolecular surface. The smaller [Symbol: see text]k'[Symbol: see text] value suggested a larger entropy content in the D2O case due to increased sampling of macromolecular conformational substates. PMID- 18286277 TI - Use of normal modes for structural modeling of proteins: the case study of rat heme oxygenase 1. AB - We present an original approach based on full-atom normal mode analysis (NMA) aimed to expand the general framework of homology modeling. Using the rat heme free oxygenase 1 as a case system, we show how NMA can be used to model different physiologically relevant conformations of the same protein. Starting from a unique heme-bound X-ray structure, and using two structural templates corresponding to a human and an incomplete rat heme-free structures, we generate models of the rat unbound species with open and closed conformations. Less than 100 lowest frequency modes of the target were sufficient to obtain the heme-free conformations, the closest to the templates. The rat HO-1 model built for the open form shows features similar to the open form of the human heme-free oxygenase, and the one built for the closed form was similar to the incompletely resolved X-ray structure of the same protein available in the Protein DataBank. In the latter case, the use of NMA was particularly useful since it allowed to build a complete structure and therefore to discuss on the reason of the structural differences between open and closed forms. This study shows that the amount of main chain flexibility provided by the normal modes can lead to major improvements in homology modeling approaches. Such applications will allow the characterization of alternative conformations of a target protein with respect to the templates and/or the construction of good quality 3D models based on existing templates with unresolved parts in their tertiary structure. PMID- 18286278 TI - Structural characterization of the exopolysaccharide PS-EDIV from Sphingomonas pituitosa strain DSM 13101. AB - Members of the bacterial genus Sphingomonas are known to produce highly viscous polysaccharides in solution. The exopolysaccharide PS-EDIV was produced by Sphingomonas pituitosa strain DSM 13101, purified using centrifugation, and precipitation and its structure was elucidated by 1D and 2D NMR techniques and chemical microderivatization combined with various mass spectrometric techniques. The following repeating unit of the polysaccharide could be identified: [Formula: see text]. In addition, the polysaccharide also contains acetyl and glyceryl groups whose exact positions were not determined. PS-EDIV is similar in structure to a known exopolysaccharide but differs in being the first bacterial polysaccharide in which two different glucuronic acids are combined. It caused a high viscosity of the culture broth after cultivation for 48 h, although a gelation was not observed. PMID- 18286279 TI - Nisin-controlled extracellular production of apidaecin in Lactococcus lactis. AB - Apidaecins are heat-stable, nonhelical antibacterial peptides isolated from lymph fluid of the honeybee (Apis mellifera). These peptides are active against a wide range of gram-negative bacteria and they are the most prominent components of the honeybee humoral defense against microbial invasion. In the present study, one isoform of apidaecin, apidaecin Ho, was expressed extracellularly in the food grade bacterium Lactococcus lactis. Results showed that expression driven by the lactococcal nisA promoter and Usp45 signal peptide resulted in efficient secretion of apidaecin in L. lactis subsp. cremoris NZ9000. Recombinant apidaecin was purified by gel filtration and semipreparative RP-HPLC, and about 10 mg active recombinant apidaecin was obtained from 1,000 ml culture. This is the first report on the nisin-controlled extracellular production of active apidaecin in L. lacits. The expression and delivery of apidaecin in the food-grade L. lactis may provide a clue to facilitate the widespread application of apidaecin in the control and prevention of gram-negative bacteria infections of human and animals. PMID- 18286280 TI - Transformation and gene replacement in the facultatively chemoheterotrophic, unicellular cyanobacterium Synechocystis sp. PCC6714 by electroporation. AB - The unicellular cyanobacterium Synechocystis sp. PCC6714 can grow not only under photoautotrophic conditions, but also under chemoheterotrophic conditions if glucose is added to the medium. This makes it useful for the study of many aspects of bioenergetic mechanisms. In contrast to its closely related strain Synechocystis sp. PCC6803, which cannot grow chemoheterotrophically, Synechocystis PCC6714 is not naturally transformable. To enable gene transfer in this strain, we established a method for the introduction of self-replicating IncQ plasmids and for gene replacement using electroporation. PMID- 18286281 TI - Plantar fasciitis and calcaneal spur formation are associated with abductor digiti minimi atrophy on MRI of the foot. AB - OBJECTIVE: To determine the association of atrophy of the abductor digiti minimi muscle (ADMA), an MRI manifestation of chronic compression of the inferior calcaneal nerve suggesting the clinical diagnosis of Baxter's neuropathy, with MRI markers of potential etiologies, including calcaneal spur formation, plantar fasciitis, calcaneal edema, Achilles tendinosis and posterior tibial tendon dysfunction (PTTD). MATERIALS AND METHODS: Prevalence of calcaneal spur formation, plantar fasciitis, calcaneal edema, Achilles tendinosis and PTTD was assessed retrospectively on 100 MRI studies with ADMA and 100 MRI studies without ADMA. Patients ranged in age from 10-92 years. Pearson chi-square analyses and Fisher's exact test were used to compare prevalence of the above findings in patients with and without ADMA. Logistic regression was used to determine which variables were significantly associated with ADMA. RESULTS: Among patients with ADMA, there was significantly greater age (57.2 years vs 40.8 years, p<0.001), presence of Achilles tendinosis (22.0% vs 3.0%, P<0.001), calcaneal edema (15.0% vs 3.0%, P=0.005), calcaneal spur (48.0% vs 7.0%, P<0.001), plantar fasciitis (52.5% vs 11.0%, P<0.001), and PTTD (32.0% vs 11.0%, P<0.001). After multivariate logistic regression analysis, only age [odds ratio (OR) 1.06, 95% confidence interval (CI) 1.03, 1.09], calcaneal spur (OR 3.60, 95% CI 1.28, 10.17), and plantar fasciitis (OR 3.35, 95% CI 1.31, 8.56) remained significant. CONCLUSION: Advancing age, calcaneal spur, and plantar fasciitis are significantly associated with ADMA. Their high odds ratios support the notion of a possible etiologic role for calcaneal spur and plantar fasciitis in the progression to Baxter's neuropathy. PMID- 18286282 TI - Detailed analysis of contrast-enhanced MRI of hands and wrists in patients with psoriatic arthritis. AB - OBJECTIVE: The objective was to perform detailed analysis of the involved soft tissues, tendons, joints, and bones in the hands and wrists of patients with psoriatic arthritis (PsA). MATERIALS AND METHODS: We reviewed 23 contrast enhanced MR imaging studies (13 hands and 10 wrists) in 10 patients with the clinical diagnosis of PsA. We obtained clinical information from medical records and evaluated images for the presence of erosions, bone marrow edema, joint synovitis, tenosynovitis, carpal tunnel, and soft tissue involvement. Two board certified musculoskeletal radiologists reviewed all images independently. Differences were resolved during a subsequent joint session. RESULTS: The average duration of disease was 71.3 months, ranging from 1 month to 25 years. Eight of the 10 wrists (80%) and 6 of the 13 hands demonstrated bone erosions. Bone marrow abnormalities were shown in 5 of the 10 wrists (50%) and 4 of the 14 hands (31%). Triangular fibrocartilage tears were seen in 6 of the 10 wrists (60%). Wrist and hand joint synovitis were present in all studies (67 wrist joints and 101 hand joints). Wrist soft tissue involvement was detected in 9 of the 10 wrists (90%) and hand soft tissue involvement was present in 12 of the 13 wrists (92%). Findings adjacent to the region of soft tissue involvement included synovitis (4 wrists) and tenosynovitis (3 wrists). Bone marrow edema adjacent to the region of soft tissue involvement was seen in one wrist. Bulge of the flexor retinaculum was seen in 4 of the 10 wrists (40%) and median nerve enhancement was seen in 8 of the 10 wrists (80%). Tenosynovitis was seen in all studies (all 10 of the hands and all 13 of the wrists). The "rheumatoid" type of distribution of bony lesions was common in our study. Interobserver agreement for various findings ranged from 83% to 100%. CONCLUSION: Contrast-enhanced MRI unequivocally demonstrated bone marrow edema, erosions, tendon and soft-tissue disease, and median nerve involvement, with good interobserver reliability in patients with PsA of the hands and wrists. Disease was more extensive in the wrists than in the hands. PMID- 18286283 TI - Oncoantigens as anti-tumor vaccination targets: the chance of a lucky strike? AB - Neoplastic transformation is a multistage process and distinct gene products of specific cell regulatory pathways are involved at each stage. Identification of genes overexpressed at a specific stage provides an unprecedented opportunity to address the immune system against antigens with a driving role in tumor progression (oncoantigens). The ERBB2 oncogene is a prototype of deregulated oncogenic protein kinase membrane receptors. Mice transgenic for rat ERBB2 (BALB neuT mice) were used in this study to identify an additional set of oncoantigens expressed at defined stages by most breast carcinomas to be used as alternatives to ERBB2-driven vaccination. To address this question, we integrated the transcription data generated by comparing preneoplastic lesions and neoplasia in BALB-neuT mice with a meta-analysis on transcription profiles generated from normal and breast tumor human specimens. Forty-six putative oncoantigens identified and prioritized according to their expression on the cell membrane or in the extra cellular space, cytoplasm and nucleus were chosen for preclinical investigation as vaccination targets. PMID- 18286284 TI - Cancer treatment: the combination of vaccination with other therapies. AB - Harnessing of the immune system by the development of 'therapeutic' vaccines, for the battle against cancer has been the focus of tremendous research efforts over the past two decades. As an illustration of the impressive amounts of data gathered over the past years, numerous antigens expressed on the surface of cancer cells, have been characterized. To this end, recent years research has focussed on characterization of antigens that play an important role for the growth and survival of cancer cells. Anti-apoptotic molecules like survivin that enhance the survival of cancer cells and facilitate their escape from cytotoxic therapies represent prime vaccination candidates. The characterization of a high number of tumor antigens allow the concurrent or serial immunological targeting of different proteins associated with such cancer traits. Moreover, while vaccination in itself is a promising new approach to fight cancer, the combination with additional therapy could create a number of synergistic effects. Herein we discuss the possibilities and prospects of vaccination when combined with other treatments. In this regard, cell death upon drug exposure may be immunogenic or non-immunogenic depending on the specific chemotherapeutics. Also, chemotherapy represents one of several options available for clearance of CD4+ Foxp3+ regulatory T cells. Moreover, therapies based on monoclonal antibodies may have synergistic potential in combination with vaccination, both when used for targeting of tumor cells and endothelial cells. The efficacy of therapeutic vaccination against cancer will over the next few years be studied in settings taking advantage of strategies in which vaccination is combined with other treatment modalities. These combinations should be based on current knowledge not only regarding the biology of the cancer cell per se, but also considering how treatment may influence the malignant cell population as well as the immune system. PMID- 18286285 TI - IL-21 induces in vivo immune activation of NK cells and CD8(+) T cells in patients with metastatic melanoma and renal cell carcinoma. AB - PURPOSE: Human interleukin-21 (IL-21) is a class I cytokine previously reported in clinical studies on immune responsive cancers. Here we report the effects of systemic IL-21 therapy on the immune system in two phase 1 trials with this novel cytokine. EXPERIMENTAL DESIGN: Recombinant IL-21 was administered by intravenous bolus injection at dose levels from 1 to 100 microg/kg using two planned treatment regimens: thrice weekly for 6 weeks (3/week); or once daily for five consecutive days followed by nine dose-free days (5 + 9). The following biomarkers were studied in peripheral blood mononuclear cells (PBMC) during treatment: phosphorylation of STAT3, alterations in the composition of leukocyte subsets, ex vivo cytotoxicity, expression of effector molecules in enriched CD8(+) T cells and CD56(+) NK cells by quantitative RT-PCR, and gene array profiling of CD8(+) T cells. RESULTS: Effects of IL-21 were observed at all dose levels. In the 5 + 9 regimen IL-21 induced a dose dependent decrease in circulating NK cells and T cells followed by a return to baseline in resting periods. In both CD8(+) T cells and CD56(+) NK cells we found up-regulation of perforin and granzyme B mRNA. In addition, full transcriptome analysis of CD8(+) T cells displayed changes in several transcripts associated with increased cell cycle progression, cellular motility, and immune activation. Finally, cytotoxicity assays showed that IL-21 enhanced the ability of NK cells to kill sensitive targets ex vivo. CONCLUSIONS: IL-21 was biologically active at all dose levels administered with evidence of in vivo NK cell and CD8(+) T cell activation. PMID- 18286286 TI - TARC and RANTES enhance antitumor immunity induced by the GM-CSF-transduced tumor vaccine in a mouse tumor model. AB - INTRODUCTION: Transduction of the granulocyte-macrophage colony stimulating factor (GM-CSF) gene into mouse tumor cells abrogates their tumorigenicity in vivo. Our previous report demonstrated that gene transduction of GM-CSF with either TARC or RANTES chemokines suppressed in vivo tumor formation. In this paper, we examined whether the addition of either recombinant TARC or RANTES proteins to irradiated GM-CSF-transduced tumor vaccine cells enhanced antitumor immunity against established mouse tumor models to examine its future clinical application. MATERIALS AND METHODS: Three million irradiated WEHI3B cells retrovirally transduced with murine GM-CSF cDNA in combination with either recombinant TARC or RANTES were subcutaneously inoculated into syngeneic WEHI3B preestablished BALB/c mice. RESULTS: Vaccinations were well tolerated. Mice treated with GM-CSF-transduced cells and the chemokines demonstrated significantly longer survival than mice treated with GM-CSF-transduced cells alone. Splenocytes harvested from mice treated with the former vaccines produced higher levels of IL-4, IL-6, IFN-gamma, and TNF-alpha, suggesting enhanced innate and adaptive immunity. Immunohistochemical analysis of tumor sections after vaccination revealed a more significant contribution of CD4+ and CD8+ T cells to tumor repression in the combined vaccine groups than controls. CONCLUSIONS: TARC and RANTES enhance the immunological antitumor effect induced by GM-CSF in mouse WEHI3B tumor models and may be clinically useful. PMID- 18286287 TI - High frequency of immature dendritic cells and altered in situ production of interleukin-4 and tumor necrosis factor-alpha in lung cancer. AB - INTRODUCTION: Antigen-presenting cells, like dendritic cells (DCs) and macrophages, play a significant role in the induction of an immune response and an imbalance in the proportion of macrophages, immature and mature DCs within the tumor could affect significantly the immune response to cancer. DCs and macrophages can differentiate from monocytes, depending on the milieu, where cytokines, like interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) induce DC differentiation and tumor necrosis factor (TNF)-alpha induce DC maturation. Thus, the aim of this work was to analyze by immunohistochemistry the presence of DCs (S100+ or CD1a+), macrophages (CD68+), IL-4 and TNF-alpha within the microenvironment of primary lung carcinomas. RESULTS: Higher frequencies of both immature DCs and macrophages were detected in the tumor-affected lung, when compared to the non-affected lung. Also, TNF-alpha positive cells were more frequent, while IL-4-positive cells were less frequent in neoplastic tissues. This decreased frequency of mature DCs within the tumor was further confirmed by the lower frequency of CD14-CD80+ cells in cell suspensions obtained from the same lung tissues analyzed by flow cytometry. CONCLUSION: These data are discussed and interpreted as the result of an environment that does not oppose monocyte differentiation into DCs, but that could impair DC maturation, thus affecting the induction of effective immune responses against the tumor. PMID- 18286288 TI - Resveratrol analog trans 3,4,5,4'-tetramethoxystilbene (DMU-212) mediates anti tumor effects via mechanism different from that of resveratrol. AB - PURPOSE: Resveratrol is a well-known chemopreventive and chemotherapeutic agent. Among all of the resveratrol analogs synthesized, 3,4,5,4'-tetramethoxystilbene (DMU-212) shows high activity and selectivity against various cancer cell types. The objective of this study is to investigate why DMU-212 has higher anti-tumor activity than resveratrol. METHODS: The effects of DMU-212 and resveratrol on cell viability, cell cycle, Stat3 activation, and microtubule dynamic were investigated and compared using MTT assay, cell cycle analysis, Western blot, tubulin polymerization assay, respectively, in MDA-MB-435 and MCF-7 human breast cancer cells. RESULTS: Compared to resveratrol, DMU-212 exerted a significantly higher growth inhibition in both cell lines. Further studies demonstrated that DMU-212 acted via different mechanisms from resveratrol. First, DMU-212 induced predominantly G2/M arrest whereas resveratrol induced G0/G1 arrest in both cell lines. Correlating with these findings, resveratrol induced more dramatic changes in the expression of Cyclin D1 compared to DMU-212. Second, DMU-212 induced apoptosis and reduced the expression of multiple anti-apoptotic proteins more appreciably than resveratrol. Third, while both agents inhibited Stat3 phosphorylation, treatments of DMU-212 but not resveratrol led to a significant increase in tubulin polymerization. The higher sensitivity to DMU-122 in MDA-MB 435 correlated with the more prominent effects seen in these parameters in this cell line, as compared to MCF7. CONCLUSION: Compared to resveratrol, the novel stilbene derivative, DMU-212, had higher anti-tumor effects, which are likely owing to its modulation of multiple cellular targets. PMID- 18286289 TI - Radiofrequency ablation of benign thyroid nodules: safety and imaging follow-up in 236 patients. AB - This study evaluated the safety and volume reduction of ultrasonography (US) guided radiofrequency ablation (RFA) for benign thyroid nodules, and the factors affecting the results obtained. A total of 302 benign thyroid nodules in 236 euthyroid patients underwent RFA between June 2002 and January 2005. RFA was carried out using an internally cooled electrode under local anesthesia. The volume-reduction ratio (VRR) was assessed by US and safety was determined by observing the complications during the follow-up period (1-41 months). The correlation between the VRR and several factors (patient age, volume and composition of the index nodule) was evaluated. The volume of index nodules was 0.11-95.61 ml (mean, 6.13 +/- 9.59 ml). After ablation, the volume of index nodules decreased to 0.00-26.07 ml (mean, 1.12 +/- 2.92 ml) and the VRR was 12.52 100% (mean, 84.11 +/- 14.93%) at the last follow-up. A VRR greater than 50% was observed in 91.06% of nodules, and 27.81% of index nodules disappeared. The complications encountered were pain, hematoma and transient voice changes. In conclusion, RFA is a safe modality effective at reducing volume in benign thyroid nodules. PMID- 18286290 TI - Visualization of antigen-specific human cytotoxic T lymphocytes labeled with superparamagnetic iron-oxide particles. AB - New technologies are needed to characterize the migration and survival of antigen specific T cells in vivo. In this study, we developed a novel technique for the labeling of human cytotoxic T lymphocytes with superparamagnetic iron-oxide particles and the subsequent depiction with a conventional 1.5-T magnetic resonance scanner. Antigen-specific CD8(+) T lymphocytes were labeled with ferucarbotran by lipofection. The uptake of ferucarbotran was confirmed by immunofluorescence microscopy using a dextran-specific antibody, and the intracellular enrichment of iron was measured by atomic absorption spectrometry. The imaging of T cells was performed by magnetic resonance on day 0, 2, 7 and 14 after the labeling procedure. On day 0 and 2 post labeling, a pronounced shortening of T2*-relaxation times was observed, which diminished after 7 days and was not detectable anymore after 14 days, probably due to the retained mitotic activity of the labeled T cells. Of importance, the antigen-specific cytolytic activity of the T cells was preserved following ferucarbotran labeling. Efficient ferucarbotran labeling of functionally active T lymphocytes and their detection by magnetic resonance imaging allows the in vivo monitoring of T cells and, subsequently, will impact the further development of T cell-based therapies. PMID- 18286291 TI - Coronary 64-slice CT angiography predicts outcome in patients with known or suspected coronary artery disease. AB - The aim of this study was to assess the prognostic value of 64-slice CT angiography (CTA) in patients with known or suspected coronary artery disease (CAD). Sixty-four-slice coronary CTA was performed in 220 patients [mean age 63 +/- 11 years, 77 (35%) female] with known or suspected CAD. CTA images were analyzed with regard to the presence and number of coronary lesions. Patients were followed-up for the occurrence of the following clinical endpoints: death, nonfatal myocardial infarction, unstable angina, and coronary revascularization. During a mean follow-up of 14 +/- 4 months, 59 patients (27%) reached at least one of the predefined clinical endpoints. Patients with abnormal coronary arteries on CTA (i.e., presence of coronary plaques) had a 1st-year event rate of 34%, whereas in patients with normal coronary arteries no events occurred (event rate, 0%, p < 0.001). Similarly, obstructive lesions (> or =50% luminal narrowing) on CTA were associated with a high first-year event rate (59%) compared to patients without stenoses (3%, p < 0.001). The presence of obstructive lesions was a significant independent predictor of an adverse cardiac outcome. Sixty-four-slice CTA predicts cardiac events in patients with known or suspected CAD. Conversely, patients with normal coronary arteries on CTA have an excellent mid-term prognosis. PMID- 18286292 TI - Dermal fibrosis in male pattern hair loss: a suggestive implication of mast cells. AB - A relationship has been suggested between mast cells (MCs) and male pattern hair loss (MPHL), because of histological evidence of perifollicular fibrosis and increased mast cell numbers. Two paired punch biopsies were taken from balding vertexes and non-balding occipital promontory areas of ten patients with MPHL (Ludwig-Hamilton IIIv to IV) and from five normal subjects aged from 20 to 35 years. Masson trichrome and Victoria blue staining were performed to observe collagen frameworks and elastic fiber structures. Numbers of immunoreactive MCs stained with anti-tryptase or anti-chymase antibody were counted. It was found that collagen bundles were significantly increased in balding vertexes than in non-balding occiput scalp skin. A near 4-fold increase in elastic fibers was observed in both vertex and occiput scalp skins with MPHL versus controls. Total numbers of MCs (tryptase-positive) in site-matched scalp samples were about 2 fold higher in MPHL subjects than in normal controls. Percentage elastic fiber (%) was found to be relatively well-correlated with tryptase and chymase-positive MCs. These findings suggest that accumulated MCs might be responsible for increased elastic fiber synthesis in MPHL, and indicate that future investigations are warranted. PMID- 18286293 TI - Prenatal diagnosis of Ebstein's anomaly using spatio-temporal image correlation (STIC) and inversion mode. AB - BACKGROUND: Ebstein's anomaly is a rare cardiac defect where the septal and posterior leaflets are displaced, towards the right ventricle. The leaflets are dysplastic and stuck to the ventricular wall. Its antenatal diagnosis is usually made through bidimensional echocardiography, which also has prognostic value. Recently, the technological breakthrough of three-dimensional ultrasound (3D-US) offered new diagnostic tools for congenital heart defects, less dependent on the ultrasonographer experience, when compared to two-dimensional ultrasound (2D-US). The spatio-temporal image correlation (STIC) technique allows the acquisition of the fetal heart volume and its structures as a 4D cineloop sequence showing the complete cardiac cycle. Inversion mode is a new image analysis tool for the examination of fluid-filled fetal structures that inverts the gray scale. CASE REPORT: We present a case of Ebstein's anomaly diagnosed at 26 weeks of pregnancy through bidimensional echocardiography. We emphasize its main findings in 3D-US using the STIC and inversion mode techniques. PMID- 18286294 TI - Chondrosarcoma of the uterus: case report and literature review. AB - BACKGROUND: Pure heterologous sarcomas of the uterine corpus are extremely rare, accounting for 4% of all uterine sarcomas. Primary chondrosarcoma, which is characterized by the absence of epithelial or other heterologous mesenchymal elements, is included in this group. To this date, only 17 cases, including the presenting case, have been reported. CASE: A 55-year-old female presenting with post-menopausal bleeding was diagnosed with chondrosarcoma of the uterus, after abdominal hysterectomy and bilateral salpingoophorectomy. After 8 months of surgery, there is no evidence of recurrence after receiving external radiotherapy and brachytherapy. CONCLUSION: Primary chondrosarcoma of the uterus is an extremely rare uterine tumour most frequently diagnosed by the pathologist. They are usually aggressive malignant tumours with an early relapse and metastases. PMID- 18286295 TI - Neck dissection: present and future? AB - A number of issues are at the forefront of current considerations in surgical treatment of the neck in head and neck cancer. These include proposed new definitions of lymph node levels that will lend themselves to clinical and radiographic examination, the possibility of employing molecular studies to supply information on the metastatic potential of the primary tumor in the clinically negative neck, and the results of multi-institutional prospective pathologic studies of neck dissection specimens examining the early distribution of lymph node metastases from various primary sites, to design more effective and efficient surgical procedures for treatment. The pertinent current literature was reviewed, and appropriate data extracted. Various new landmarks have been defined to distinguish the boundaries between sublevels IB and IIA, the lateral borders of level VI, and the boundaries of level VII. These landmarks are more readily distinguishable on physical and radiographic examination than the definitions currently in use. Numerous molecular studies have been employed to detect subclinical metastatic deposits in the neck, but none have been found sufficiently reliable for practical application. Multi-institutional studies have shown that sublevels IIB and level IV are rarely within the first level of lymphatic drainage routes for most primary squamous cell cancers of the head and neck. Therefore, elective selective neck dissections may be further modified to reduce morbidity and operating time. Various new issues in the treatment of cervical metastatic disease are discussed in an effort to improve the accuracy of pretreatment staging, identification of occult disease, and modification of surgical treatment to optimize efficiency and reduce morbidity. PMID- 18286296 TI - Injection of intravitreal bevacizumab (Avastin) as a preoperative adjunct before vitrectomy surgery in the treatment of severe proliferative diabetic retinopathy (PDR). AB - PURPOSE: To evaluate the use of preoperative intravitreal bevacizumab (IVB) in patients undergoing pars plana vitrectomy (PPV) for complications of proliferative diabetic retinopathy (PDR). METHODS: We studied 22 patients with severe PDR. A preoperative complexity score (CS) was recorded. Eleven eyes were treated with IVB, 1.25 mg, 5-7 days before PPV (group 1), and 11 eyes underwent direct PPV (group 2). Surgical time and intra-operative manoeuvres were recorded. Main outcome measure was feasibility of surgery, secondary goal was the visual and anatomic outcome at 6 months. RESULTS: The average CS was 5.5, and was similar in the two groups. Mean surgical time was 57 minutes in group 1 vs 83 minutes in group 2; mean tool exchanges was 27 vs 53, intraoperative bleeding 5 vs 15, endodiathermy 2 vs 9. No complications were recorded after IVB. Mean pre operative BCVA was 1.87 logMAR in group 1 and logMAR 2.04 in group 2. Mean pre operative BCVA was 1.87 logMAR in the bevacizumab group and 2.04 logMAR in group 2, not significantly different (p = 0.7). Mean post-operative BCVA at 6 months was 0.88 logMAR in group 1 and logMAR 2.01 in control group 2, significantly different (p = 0.01). Post-operative BVCA improved in bevacizumab group from pre operative value (p = 0.15), while in control group there was non-significant increase (p = 0.96). Anatomical attachment was achieved in 11 patients in group 1 vs nine patients in group 2. CONCLUSIONS: IVB administered prior to vitrectomy was well tolerated and reduced active neovascularization, thus facilitating PPV. PMID- 18286297 TI - Microarray-based gene expression analysis during retinal maturation of albino rats. AB - BACKGROUND: In recent years, the rat has become a commonly-used animal model for the study of retinal diseases. Similar to other tissues, the retina undergoes significant functional changes during maturation. Aiming to gain knowledge on additional aspects of retinal maturation, we performed gene expression and histological analyses of the rat retina during maturation. METHODS: Rat retinas were dissected at three time points. Histological examination of the samples was performed, and the expression levels of retinal genes were evaluated using the rat whole-genome microarray system. Quantitative real-time PCR analysis was used to validate selected expression patterns. Various statistical and bioinformatic tools were used to identify differentially expressed genes. RESULTS: The microarray analysis revealed a relatively high number of highly expressed non annotated genes. We identified 603 differentially expressed genes, which were grouped into six clusters based on changes in expression levels during the first 20 weeks of life. A bioinformatic analysis of these clusters revealed sets of genes encoding proteins with functions that are likely to be relevant to retinal maturation (potassium, sodium, calcium, and chloride channels, synaptic vesicle transport, and axonogenesis). The histological analysis revealed a significant reduction of outer nuclear layer thickness and retinal ganglion cell number during maturation. CONCLUSIONS: These data, taken together with our previously reported electrophysiological data, contribute to our understanding of the retinal maturation processes of this widely-used animal model. PMID- 18286298 TI - Macular translocation surgery: computer simulation of visual perception. AB - BACKGROUND: Macular translocation can be associated with visual improvement, but patients often experience symptoms of confusion or diplopia. There is a high incidence of suppression of the operated or the fellow eye. The aim of this study is to use computer software to examine the pre- and post-operative fundal images, in order to better understand how patients see after macular translocation surgery. METHODS: We created a graphical user interface that allowed a user to identify and record common landmark points in pre- and post-operative fundal images. We used these points to carry out interpolations using two algorithms, namely bilinear and thin-plate spline transformations. The transformations were applied to the Mona Lisa in order to appreciate how patients might see. RESULTS: Given two sets of corresponding points, both algorithms were able to approximate the effect of the surgery. Bilinear transformation was able to account for changes to the retina as a whole, including rotation, stretches, compression and shear. The thin-plate spline algorithm additionally accounted for the considerable regional and uneven local effects. Applying the later algorithm to the Mona Lisa produced inconsistent and warped images. CONCLUSIONS: Our results confirmed that neurosensory redistribution was associated with most cases of MT360. We infer from these results that corresponding retinal elements between two eyes would no longer correspond after surgery. The distortion of images from the operated eye could not be completely corrected by squint surgery, and this may account for the high incidence of suppression of the fellow or the operated eye after surgery. PMID- 18286299 TI - Central corneal thickness and Diaton transpalpebral tonometry. AB - BACKGROUND: To examine the effects of central corneal thickness on the measures obtained from transpalpebral tonometry (Diaton), and to identify correlations between intraocular pressure (IOP) measurements with Diaton and the Goldmann applanation tonometer (GAT). METHODS: In this cross-sectional study, 162 eyes of 81 participants were included. Intraocular pressure measurements were obtained in all patients using Diaton and GAT. Central corneal thickness was determined by ultrasound pachymetry. The participants were stratified by corneal thickness: group I <530 microm (n = 56), group II 530-560 microm (n = 65), and group III >560 microm (n = 41). RESULTS: There were moderate correlations between IOP readings obtained using the Diaton and corrected GAT (C-GAT) (r = 0.303; P < 0.0001), and between corrected Diaton (C-Diaton), and C-GAT (r = 0.399; P < 0.0001). The mean Diaton tonometer readings were lower than C-GAT measurements (Diaton-corrected GAT mean difference, 0.9 +/- 3.8 mmHg; c-Diaton-corrected GAT mean difference, 0.7 +/- 3.5 mmHg). Differences were detected between the groups of patients for the GAT values [2.4 +/- 3.6 mmHg for those with the thinnest corneas (<530 microm), 0.7 +/- 3.6 mmHg for those with moderate corneas (between 531 microm and 560 microm), and -0.6 +/- 3.6 mmHg for those with the thickest (>560 microm) corneas], whereas a significantly lower difference (0.9 +/- 3.8 mmHg) was noted for the Diaton values of all individuals. CONCLUSIONS: The Diaton measurements show moderate correlation with those provided by applanation tonometry. The Diaton tonometer seems to be more affected by the corneal thickness, especially in the thinnest corneas. PMID- 18286300 TI - Ischemic colitis: analysis of risk factors for postoperative mortality. AB - BACKGROUND: Ischemic colitis is a disease with high postoperative mortality when surgery is necessary. The definition of risk factors for perioperative mortality, which is currently lacking in the literature, could be helpful in clinical decision making and in optimizing perioperative treatment. MATERIALS AND METHODS: Based on a prospective database, 85 consecutive patients undergoing surgery for ischemic colitis between November 04, 2001 and October, 26, 2004 at the Department of Surgery, University of Heidelberg, were included in this study. The influence of different known factors on perioperative mortality such as age, type of operation, blood loss, comorbidities, hospital course, and complications was tested by univariate and multivariate analysis. RESULTS: Sixty-seven percent of patients were operated as emergency cases (within 24 h after surgical evaluation). About half of the patients underwent subtotal or total colectomy and 80% had stoma creation. Twenty-two percent of patients developed surgical complications and 47% of patients died in the further postoperative course. Univariate analysis showed underlying cardiovascular diseases, American Society of Anesthesiologists (ASA) status, emergency surgery, total colectomy, elevated intraoperative blood loss and intraoperative allogeneic blood transfusion or transfusion of fresh frozen plasma to be associated with an increased postoperative mortality. Multivariate analysis confirmed ASA status > III, emergency surgery, and blood loss to be independently associated with postoperative mortality in ischemic colitis. CONCLUSIONS: The mortality of patients requiring surgery for ischemic colitis will remain high as the majority of afflicted patients are patients with significant comorbidities in a reduced general condition. But earlier diagnosis and measures to reduce blood loss may contribute to improving the overall outcome. PMID- 18286301 TI - Symptom-focused results after laparoscopic fundoplication for refractory gastroesophageal reflux disease--a prospective study. AB - BACKGROUND AND AIMS: Symptoms of gastroesophageal reflux disease (GERD) are common in the general population. Although the results of laparoscopic fundoplication are well documented, there have been no reports on the operative outcome in patients refractory to or with only partial response to medical therapy for GERD. PATIENTS-METHODS: Thirty-two patients with GERD, whose continuous high doses of medical treatment with proton-pump inhibitors produced no or only partial symptom relief, underwent laparoscopic Nissen fundoplication. Symptoms were evaluated with a standardized questionnaire preoperatively and 12 months after surgery. RESULTS: The complete follow-up evaluation was obtained in 30 out of the 32 patients. The main symptoms before surgery were regurgitation (93%), heartburn (60%), epigastric pain (47%), and globus sensation (47%). All patients were relieved from heartburn, vomiting, and globus sensation. Dysphagia was relieved in 75% of the patients and regurgitation in 86%. Dysphagia as a new symptom occurred in 9%. The overall morbidity rate was 16%. Patient satisfaction rate was 87%. CONCLUSION: Laparoscopic fundoplication seems to be an effective treatment for severe, drug-resistant GERD. The high patient satisfaction rate and the positive therapeutic response in 95% of patients justify this procedure in this strictly selected group of patients. PMID- 18286302 TI - Localization of the ergtoxin-1 receptors on the voltage sensing domain of hERG K+ channel by AFM recognition imaging. AB - The inhibition of the human ether-a-go-go-related (hERG) K+ channels is the major cause of long QT syndromes inducing fatal cardiac arrhythmias. Ergtoxin 1 (ErgTx1) belongs to scorpion-toxins, which are K+ channel-blockers, and binds to hERG channel with 1:1 stoichiometry and high affinity (Kd approximately 10 nM). Nevertheless, patch-clamp recordings recently demonstrated that ErgTx1 does not establish complete blockade of hERG currents, even at high ErgTx1 concentrations. Such phenomenon is supposed to be consistent with highly dynamic conformational changes of the outer pore domain of hERG. In this study, simultaneous topography and recognition imaging (TREC) on hERG HEK 293 cells was used to visualize binding sites on the extracellular part of hERG channel (on S1-S2 region) for Anti-Kv11.1 (hERG-extracellular-antibody). The recognition maps of hERG channels contained recognition spots, haphazardly distributed and organized in clusters. Recognition images after the addition of ErgTx1 at high concentrations ( approximately 1 microM) revealed subsequent partial disappearance of clusters, indicating that ErgTx1 was bound to the S1-S2 region. These results were supported by AFM force spectroscopy data, showing for the first time that voltage sensing domain (S1-S4) of hERG K+ channel might be one of the multiple binding sites of ErgTx1. PMID- 18286304 TI - A variant or a "new" postaxial acrofacial dysostosis syndrome. AB - In this paper, we report a boy with remarkable phenotype and noteworthy radiographic abnormalities. He presented with distinctive facies, mesomelic shortening and asymmetry of the extremities, symmetrical ulnar and fibular ray absence in the hands and feet, with unique hypoplastic/dysplastic radiographic abnormalities. His mental development was normal. We believe that he represents a previously unreported form of the postaxial acrofacial dysostosis syndrome. PMID- 18286303 TI - Episomal amplification of MYCN in a case of medulloblastoma. AB - Gene amplification, in the form of double minutes (dmin) and/or homogeneously staining regions (hsr), is frequently associated with tumor development. A well known example is neuroblastoma for which MYCN gene (v-myc myelocytomatosis viral related oncogene) amplification has a relevant prognostic significance. A third cryptic form of amplification, cytogenetically invisible and composed of episomes, has been also described, but it is very rarely seen in primary tumors. In this paper, we report on MYCN amplification, in the form of episomes, in a case of medulloblastoma. Detailed fluorescence in situ hybridization and real time quantitative polymerase chain reaction analyses revealed an amplified genomic segment of approximately 590 kb containing only the genes MYCN and N-cym (v-myc myelocytomatosis viral-related oncogene, neuroblastoma-derived opposite strand). To the best of our knowledge, this is the first report of a solid primary tumor showing MYCN amplification in the form of episomes. PMID- 18286305 TI - Growth and development rates in a riparian spider are altered by asynchrony between the timing and amount of a resource subsidy. AB - Rapid growth in response to increased prey abundance may be induced by environmental variability associated with resource subsidies. Spiders living in riparian areas are subject to frequent, episodic bursts of aquatic prey (subsidies). These periods of high resource abundance may occur at different points in recipient consumers' development through variation in emergence patterns of prey between years or across a landscape. We examine how variable timing of subsidy abundance intersects with life history scheduling to produce different growth and development outcomes for individuals within a population. Through a series of controlled feeding experiments, we tested the hypotheses that the spider Tetragnatha versicolor: (1) exhibits compensatory growth in response to subsidy variability, (2) that rapid increases in mass may result in a greater risk of mortality, and (3) that the timing of subsidy resources relative to the development schedule of this spider may produce different outcomes for individual growth patterns and adult condition. Spiders fed at very high rates grew fastest but also showed evidence of increased mortality risk during moulting. T. versicolor is capable of exhibiting strong growth compensation-individuals suffering initial growth restriction were able to catch up completely with animals on a constant diet utilising the same amount of food. Spiders that received an early pulse of resources (simulating an early arrival of an aquatic insect subsidy to riparian forests) did worse on all measures of development and fitness than spiders that received either a constant supply of food or a late pulse of resources. Importantly, receiving large amounts of food early in life appears to actually confer relative disadvantages in terms of later performance compared with receiving subsidies later in development. Subsidies may provide greater benefits to individuals or age cohorts encountering this resource abundance closer to the onset of reproductive efforts than subsidies arriving early in development. PMID- 18286306 TI - Renal cell carcinoma in a patient with cystinosis and inflammatory bowel disease: a case report. AB - An 18-year-old male patient with a history of cystinosis and inflammatory bowel disease, who was being treated with cysteamine and 6-mercaptopurine, had an incidental finding of a mass in the right kidney on a renal ultrasound to evaluate worsening renal insufficiency. Excisional biopsy demonstrated papillary renal cell carcinoma, Type 2. Renal cell carcinoma is typically a disease of older adults, and it is not known to be associated with cystinosis. This is the first published report of renal cell carcinoma in a patient with cystinosis. PMID- 18286307 TI - Circulating calcification inhibitors and vascular properties in children after renal transplantation. AB - Pediatric transplant patients are known to have vascular abnormalities. The calcification inhibitors matrix Gla protein (MGP) and fetuin-A play an important role in the pathophysiology of vascular calcification. In the cross-sectional study reported here, we examined the circulating levels of fetuin-A and MGP in children after renal transplantation compared to healthy children and the association of these factors with vascular properties of the carotid artery. Levels of MGP and fetuin-A together with vascular properties of the carotid artery were determined in 29 pediatric renal transplant recipients and 54 healthy controls. The level of fetuin-A was decreased in the transplant group relative to the control group (P=0.005), whereas the level of MGP (both non-phosphorylated MGP and non-carboxylated MGP) did not differ between groups. The intima-media thickness (P<0.001) and the elasticity (P=0.002) of the carotid artery were significantly increased in children after renal transplantation compared to healthy children. No associations between vascular parameters and calcification inhibitors were found in either group. Circulating levels of MGP and fetuin-A could not be identified as independent predictors of vascular stiffness or other carotid artery parameters in pediatric renal transplant recipients. Future prospective studies in pediatric ESRD and transplant patients are needed to learn more about the role of calcification inhibitors in relation to the prevention of vascular damage. PMID- 18286308 TI - Long-term follow-up of renal function in patients after surgery for obstructive uropathy. AB - In this prospective study, selected biochemical markers of glomerular and tubular function, proteinuria, and ultrasound findings in 62 pediatric patients who underwent surgery for obstructive uropathy were examined. Patients were younger than 12 months, normocreatininemic at the time of surgery, and examined at a mean age of 6.3+/-0.9 years. Out of the markers tested, serum concentration of cystatin C was significantly higher in patients when compared with the control group (p<0.001), and serum creatinine concentration was within reference interval in all patients. With respect to tubular function, 26% of patients had decreased concentration ability. Proteinuria was detected in 4.8% of patients. On ultrasound, 66.7% of kidneys after surgery had residual dilatation of the renal pelvis. The patients thrive well, and their somatic parameters do not differ from their peers. Half of the patients had one or more urinary tract infections from the date of surgery to the date of examination. Study results support the need for long-term nephrologic follow-up in patients after surgery for obstructive uropathy. The hypothesis that renal function in patients undergoing surgery aged younger than 3 or 6 months is better when compared with those aged 6 to 12 months has not been confirmed. PMID- 18286309 TI - Kidney cysts, pancreatic cysts, and biliary disease in a mouse model of autosomal recessive polycystic kidney disease. AB - Mutations in PKHD1 cause autosomal recessive polycystic kidney disease (ARPKD). We produced a mouse model of ARPKD by replacing exons 1-3 of Pkhd1 with a lacZ reporter gene utilizing homologous recombination. This approach yielded heterozygous Pkhd1 (lacZ/+) mice, that expressed beta-galactosidase in tissues where Pkhd1 is normally expressed, and homozygous Pkhd1 (lacZ/lacZ) knockout mice. Heterozygous Pkhd1 (lacZ/+) mice expressed beta-galactosidase in the kidney, liver, and pancreas. Homozygous Pkhd1 (lacZ/lacZ) mice lacked Pkhd1 expression and developed progressive renal cystic disease involving the proximal tubules, collecting ducts, and glomeruli. In the liver, inactivation of Pkhd1 resulted in dilatation of the bile ducts and periportal fibrosis. Dilatation of pancreatic exocrine ducts was uniformly seen in Pkhd1 (lacZ/lacZ ) mice, with pancreatic cysts arising less frequently. The expression of beta-galactosidase, Pkd1, and Pkd2 was reduced in the kidneys of Pkhd1 (lacZ/lacZ ) mice compared with wild-type littermates, but no changes in blood urea nitrogen (BUN) or liver function tests were observed. Collectively, these results indicate that deletion of exons 1-3 leads to loss of Pkhd1 expression and results in kidney cysts, pancreatic cysts, and biliary ductal plate malformations. The Pkhd1 (lacZ/lacZ ) mouse represents a new orthologous animal model for studying the pathogenesis of kidney cysts and biliary dysgenesis that characterize human ARPKD. PMID- 18286311 TI - Molecular dynamics simulations on hydrogen adsorption in finite single walled carbon nanotube bundles. AB - Molecular dynamics simulations of the adsorption of hydrogen molecules in finite single-walled carbon nanotube bundles are presented using a curvature dependent force field. The heat of formation and the effective adsorption capacity are expressed as a function of H(2) distance from adsorbent. The heat of adsorption decreases rapidly with the distance and increasing H(2) loading results in weakening adsorption strength. The effects of nanotube packing and bundle thickness on hydrogen adsorption strength were investigated and the results show that the heat of adsorption can be improved slightly if hydrogen molecules are placed in thicker and inhomogeneously packed nanotube bundles. Only very small diameter nanotube bundles were found to hold promise for significant hydrogen storage for onboard applications. PMID- 18286310 TI - DFT-based molecular modeling and vibrational study of the La(III) complex of 3,3' (benzylidene)bis(4-hydroxycoumarin). AB - Molecular modeling of the La(III) complex of 3,3'-(benzylidene)bis(4 hydroxycoumarin) (PhDC) was performed using density functional theory (DFT) methods at B3LYP/6-31G(d) and BP86/TZP levels. Both Stuttgart-Dresden effective core potential and ZORA approximation were applied to the La(III) center. The electron density distribution and the nucleophilic centers of the deprotonated ligand PhDC(2-) in a solvent environment were estimated on the basis of Hirshfeld atomic charges, electrostatic potential values at the nuclei, and Nalewajski Mrozek bond orders. In accordance with the empirical formula La(PhDC)(OH)(H(2)O), a chain structure of the complex was simulated by means of two types of molecular fragment: (1) two La(III) cations bound to one PhDC(2-) ligand, and (2) two PhDC(2-) ligands bound to one La(III) cation. Different orientations of PhDC(2-), OH(-) and H(2)O ligands in the La(III) complexes were investigated using 20 possible [La(PhDC(2-))(2)(OH)(H(2)O)](2-) fragments. Energy calculations predicted that the prism-like structure based on "tail-head" cis-LML2 type binding and stabilized via HO...HOH intramolecular hydrogen bonds is the most probable structure for the La(III) complex. The calculated vibrational spectrum of the lowest energy La(III) model fragment is in very good agreement with the experimental IR spectrum of the complex, supporting the suggested ligand binding mode to La(III) in a chain structure, namely, every PhDC(2-) interacts with two La(III) cations through both carbonylic and both hydroxylic oxygens, and every La(III) cation binds four oxygen atoms of two different PhDC(2-). PMID- 18286312 TI - Exploring the binding pocket for pyridopyrimidine ligands at the CCK1 receptor by molecular docking. AB - Pyridopyrimidine-based analogues are among the most highly potent and selective antagonists of cholecystokinin receptor subtype-1 (CCK1R) described to date. To better understand the structural and chemical features responsible for the recognition mechanism, and to explore the binding pocket of these compounds, we performed automated molecular docking using GOLD2.2 software on some derivatives with structural diversity, and propose a putative binding conformation for each compound. The docking protocol was guided by the key role of the Asn333 residue, as revealed by site directed mutagenesis studies. The results suggest two putative binding modes located in the same pocket. Both are characterized by interaction with the main residues revealed by experiment, Asn333 and Arg336, and differ in the spatial position of the Boc-Trp moiety of these compounds. Hydrophobic contacts with residues Thr117, Phe107, Ile352 and Ile329 are also in agreement with experimental data. Despite the poor correlation obtained between the estimated binding energies and the experimental activity, the proposed models allow us to suggest a plausible explanation of the observed binding data in accordance with chemical characteristics of the compounds, and also to explain the observed diastereoselectivity of this family of antagonists towards CCK1R. The most reasonable selected binding conformations could be the starting point for future studies. Figure Superimposition of the two putative binding conformations revealed by molecular docking for pyridopyrimidine-based CCK1 antagonists. PMID- 18286313 TI - A quasispecies approach to the evolution of sexual replication in unicellular organisms. AB - This study develops a simplified model describing the evolutionary dynamics of a population composed of obligate sexually and asexually reproducing, unicellular organisms. The model assumes that the organisms have diploid genomes consisting of two chromosomes, and that the sexual organisms replicate by first dividing into haploid intermediates, which then combine with other haploids, followed by the normal mitotic division of the resulting diploid into two new daughter cells. We assume that the fitness landscape of the diploids is analogous to the single fitness-peak approach often used in single-chromosome studies. That is, we assume a master chromosome that becomes defective with just one point mutation. The diploid fitness then depends on whether the genome has zero, one, or two copies of the master chromosome. We also assume that only pairs of haploids with a master chromosome are capable of combining so as to produce sexual diploid cells, and that this process is described by second-order kinetics. We find that, in a range of intermediate values of the replication fidelity, sexually reproducing cells can outcompete asexual ones, provided the initial abundance of sexual cells is above some threshold value. The range of values where sexual reproduction outcompetes asexual reproduction increases with decreasing replication rate and increasing population density. We critically evaluate a common approach, based on a group selection perspective, used to study the competition between populations and show its flaws in addressing the evolution of sex problem. PMID- 18286314 TI - An infected arachnoid cyst in an elderly patient. PMID- 18286315 TI - Chiasmal optic neuritis following mumps parotitis. PMID- 18286316 TI - Dissociation between canal- and otolithfunction in cerebellar atrophy. PMID- 18286317 TI - Comment to the paper of Medeiros CAM, et al. (2007) J Neurol 254:459-464. PMID- 18286318 TI - Exaggerated auditory startle responses in patients with spinal cord injury. AB - OBJECTIVE: Central nervous system reorganization following spinal cord injury (SCI) may cause functional changes in the motor tracts in patients in whom increased auditory startle responses (ASRs) have been previously reported. We hypothesized that if increased ASRs in patients with incomplete SCI were due to compensatory mechanisms, these changes would be related to severity and/or localization of the lesion. METHODS: We examined ASR characteristics in 29 SCI patients and 14 age-matched healthy volunteers. Fourteen patients had incomplete and 15 complete SCI; 10 patients had cervical and 19 thoracolumbar SCI. Five auditory stimuli were applied binaurally to subjects in a sitting position, with a 5-min interstimulus interval. Surface electromyographic recordings were obtained from orbicularis oculi (OOc), sternocleidomastoid (SCM), biceps brachii (BB), and tibialis anterior (TA) muscles. RESULTS: ASR probability was significantly higher and area-under-the-curve was significantly larger in SCM and BB in patients than in controls. ASR latency was significantly shorter in SCM and BB in patients with cervical than in those with thoracolumbar SCI (p < 0.02), but there were no statistically significant differences between complete and incomplete SCI (p > 0.1). Time span since onset correlated significantly with ASR area in OOc, SCM and BB (p < 0.05). CONCLUSION: The capability of the adult central nervous system to reorganize its circuits over time for improved functionality following injury is probably the key to understanding the increased ASRs in patients with SCI. The exaggeration of the startle reflex is potentially important since it may be useful for augmenting voluntary movement in the clinical rehabilitation of patients with SCI. PMID- 18286319 TI - Virchow-Robin spaces relate to cerebral small vessel disease severity. AB - BACKGROUND AND PURPOSE: Virchow-Robin spaces (VRs) are perivascular spaces surrounding the deep perforating brain arteries. VRs dilatation is pathologic, and it could be a manifestation of cerebral small vessel disease. In the present study we assessed the relation between VRs and silent ischemic lesions in a cohort of patients with cerebral small vessel disease. METHODS: We divided dilated VRs on MRI (1.5 Tesla) into three semi-quantitative categories in 165 first ever lacunar stroke patients. We counted asymptomatic lacunar infarcts and graded white matter lesions, and compared the prevalence of vascular risk factors in different categories of VRs. We also determined independent predictors of silent ischemic lesions. RESULTS: VRs at basal ganglia level related to age, hypertension, asymptomatic lacunar infarcts, and white matter lesions. VRs at basal ganglia level predicted silent ischemic lesions (odds ratio 10.58 per higher VRs category; 95 %- confidence interval 3.40 - 32.92). CONCLUSION: Dilated VRs in the basal ganglia relate to the severity of cerebral small vessel disease and might be a manifestation of the same small vessel abnormality that causes silent ischemic lesions. This adds a role for VRs as a potential marker for small vessel disease. PMID- 18286321 TI - [Suicidality in old age]. PMID- 18286320 TI - Clinical and molecular characterisation of a Parkinson family with a novel PINK1 mutation. AB - Homozygous mutations in the PINK1 gene have been shown to cause early-onset parkinsonism. Here, we describe a novel homozygous mutation (Q126P), identified in two affected German sisters with a clinical phenotype typical for PINK1 associated parkinsonism. We analysed lactate, pyruvate, carnitine and acylcarnitine blood levels, lactate levels under exercise and in the cerebrospinal fluid, activity of respiratory chain complexes I-IV in muscle biopsies and proteasomal activity in immortalized lymphoblasts, but found no evidence for mitochondrial or proteasomal dysfunction. MR spectroscopy revealed raised myoinositol levels in the basal ganglia of both patients, reflecting possible astroglial proliferation. PMID- 18286322 TI - [Epidemiology of suicide in older persons]. AB - The suicide risk in Germany increases with age. The suicide rates follow the so called "Hungarian" pattern. Especially the percentage of older females among all suicides has increased unproportionally in recent years in relation to their percentage in the population. Every second suicide of a female is today a suicide of a female older than 60 years. The main suicide method among older persons is hanging. Suicide attempts are rare among older persons; however, the rates are increasing among the oldest age groups. The suicide attempts of older persons are committed with a higher lethality and show rarely appellative components. Suicide attempt methods are often poisoning with pharmaceuticals. In total the suicide and suicide attempt rates among elderly are underestimated due to indirect methods (passive reactions, e.g. noneating). Reasons for suicides and suicide attempts among older persons are often psychic illnesses (mostly depression), motives often include loss of partners, loss of the social network, fear of the consequences of somatic illnesses and loss of freedom of action. Thus, often the social situation of older persons with suicide attempts changed before suicide was attempted. PMID- 18286323 TI - [Transactional model of suicidal behavior in the elderly]. AB - General theories of suicidology are insufficient to explain the overproportional rate of suicidal behaviour in the elderly. The proposed transactional model represents a bio-psycho-social foundation of suicidal behavior in the elderly. The reciprocal interactions of biological, somatic, psychological and social factors in old age which may enhance suicidal behavior as a mode of problem solving behavior are discussed. PMID- 18286324 TI - [Sexuality of suicidal elderly]. AB - Sexuality and suicidality in the elderly seem to underlie a taboo in science and in therapeutic practice. It is urgent to find an understanding about both aspects of human experience and behaviour by explorative studies. In a sample (N=67), the narratives of 56 persons (suicidal=43 and non-suicidal=23) on questions concerning their sexuality were transcribed and ideal types were developed with the systematic qualitative method of forming types by understanding. Four of these types consist of clearly more suicidal persons, in two the non-suicidals predominate, the rest is more heterogeneous in this aspect. Some of these types can be interpreted unequivocally. One type was surprisingly clearly suicidal dynamic, while others were a "female narcissistic" or a "male narcissistic" type. This explorative typology may help the interested therapist/counsellor as a landmark to relate to sexuality as an important aspect of life, even in old age, and to use the discussion of sexuality to improve the understanding of patients, especially in suicidal crisis. PMID- 18286325 TI - [Crisis intervention with elderly people]. AB - This paper gives an overview about the most important aspects of crisis intervention, with special emphasis on crisis intervention with elderly people. First a review of the development of crisis intervention is given, including of some of the major concepts, with particular emphasis on psychoanalytic aspects of crisis intervention. Then a clinical case example of a crisis intervention with an elderly woman following a suicide attempt is given and discussed. The focus lies on the description of the transference-countertransference relationship, with attempts of pressing the therapist to comply with superficial, denying and minimizing fantasies. Peculiarities of crisis intervention with elderly people are highlighted: it is necessary to emphasize that elderly people are underrepresented in most crisis services, whereby they represent the group with the highest suicide risk. Peculiarities of elderly people still are not sufficiently met and they are created by a particularly wide range of aspects. PMID- 18286326 TI - [Attitudes toward one's own aging: a question of age or a question of resources?]. AB - Recent studies have demonstrated that positive and negative stereotypes and attitudes toward age and aging may have substantial effects on the longterm development of cognitive and physical performance in old age. Relatively little is known to which degree attitudes toward one's own aging depend on (a) the current level of individual resources, (b) changes in the level of individual resources, or (c) the increasing age of older persons. The present study examined the relations between individual physical, mental, and cognitive resources and the attitudes toward one's own aging. The study was conducted with N=500 participants of the Interdisciplinary Longitudinal Study of Adult Development (ILSE) born between 1930 and 1932 (age: M=63.0, SD=0.9 at T1 and M=66.9, SD=0.9 at T2). The results indicate that the attitudes toward one's own aging (a) become more negative with increasing age, (b) most strongly depend on the available resources, (c) changes in level of resources contribute to the prediction of the attitudes toward one's own aging, and (d) age acts more as a reinforcing factor on the relation between resources and attitudes than as an independent factor on the attitudes toward one's own aging. PMID- 18286327 TI - [Impact of vascular pathology on survival times of 173 dementia patients- Hachinski's ischemic score as a predictive tool for clinical purposes]. AB - Alzheimer's disease and vascular dementia still may be looked upon as distinct nosologic entities, representing the two main etiologic categories of senile dementia. However, rather recent findings suggest a comorbidity of neurodegenerative and ischemic pathology in a majority of dementia cases in later life. The effect of the vascular pathology on the survival time was studied in 173 dementia outpatients. For 147 patients with complete datasets, we were able to gain information concerning their survival time. As an indicator of cerebrovascular morbidity the 18-point ischemic scale of Hachinski (HIS) was correlated with the survival time. Thus, we did not use the HIS for its original purpose to differentiate between degenerative and vascular dementia, but to roughly evaluate the cerebrovascular impact in a continuum model. Using the Cox model we calculated mortality risks for every point on the HIS. We found a Cox hazard ratio of 1.038 for each supplementary point on the HIS, which equals a 3.8% higher relative mortality risk. The result misses significance (p=0.092), but indicates a clear tendency towards a shortening of survival time by vascular comorbidity. Future prospective studies should integrate brain imaging to further corroborate our findings. PMID- 18286328 TI - Health: support provided and received in advanced old age. A five-year follow-up. AB - While research focuses mainly on support provided to the elderly, this paper deals with the very old as a support provider to his family as much as a care recipient from both his family and a formal network. We hypothesize that elders with declining health will try to maintain the provision of services, even when they require and receive help.A total of 340 octogenarians from the Swiss Interdisciplinary Longitudinal Study on the Oldest Old (SWILSOO) were interviewed up to five times over five years (N=1225 interviews). A multilevel model was applied to assess the effects of health, controlled for socio-demographic and family network variables, on the frequency of services that the old persons provided to their family and received from their family and formal networks. Health is operationalized in three statuses: ADL-dependent, ADL-independent frail, and robust.While the recourse to the informal network increased progressively with the process of frailty, the recourse to the formal network drastically increased for ADL-dependent individuals. Being ADL-dependent seriously altered the capacity to provide services, but ADL-independent frail persons were providers with the same frequency as the robust oldest old, showing their ability to preserve a principle of reciprocity in their exchanges with their family network. This continuity of roles may help frail persons to maintain their self-esteem and well-being. PMID- 18286331 TI - Native grasses as a management alternative on vegetated closure caps. AB - Capped waste sites often are vegetated with commercial turf grasses to increase evapotranspiration and prevent erosion and possible exposure of the barrier. Fertilizer, frequent watering, and mowing may be required to establish the turf grass and prevent invasion by trees and shrubs. Oldfield vegetation of grasses and forbs is a possible sustainable alternative to turf grass communities. To determine if oldfield vegetation can establish on caps, we (1) compared establishment of a dominant oldfield grass and a commercial turf grass under different combinations of new closure cap management: spring or summer planting and presence or absence of amendments to alleviate drought (watering, mulch) or increase soil fertility (fertilizer, lime, a nitrogen-fixing legume); (2) surveyed existing caps to determine if oldfield species establish naturally; and (3) performed a greenhouse experiment to compare growth of two native grasses under low and amended (added water, soil nutrients) conditions. Both the commercial grass and oldfield species established under new cap conditions; fertilizer, water, and mulch improved vegetation establishment in spring or summer, but legumes decreased grass cover. In the greenhouse, both native grasses grew best with amendments; however, substantial stem and root length were obtained with no fertilizer and only once-weekly watering. Existing vegetated caps supported planted grasses and naturally established oldfield species. Overall, the results indicate native grasses can establish on new caps and oldfields can serve as a management model; further work is needed to determine the management strategy to maintain herbaceous vegetation and slow woody species invasion. PMID- 18286333 TI - BSIR abstracts 2007. PMID- 18286334 TI - Genotype X diet interactions in mice predisposed to mammary cancer. I. Body weight and fat. AB - High dietary fat intake and obesity may increase susceptibility to certain forms of cancer. To study the interactions of dietary fat, obesity, and metastatic mammary cancer, we created a population of F(2) mice cosegregating obesity QTL and the MMTV-PyMT transgene. We fed the F(2) mice either a very-high-fat or a matched-control-fat diet and measured growth, body composition, age at mammary tumor onset, tumor number and severity, and formation of pulmonary metastases. SNP genotyping across the genome facilitated analyses of QTL and QTL x diet interaction effects. Here we describe development of the F(2) population (n = 615) which resulted from a cross between the polygenic obesity model M16i and FVB/NJ-TgN (MMTV-PyMT)(634Mul), effects of diet on growth and body composition, and QTL and QTL x diet and/or gender interaction effects for growth and obesity related phenotypes. We identified 38 QTL for body composition traits that were significant at the genome-wide 0.05 level, likely representing nine distinct loci after accounting for pleiotropic effects. QTL x diet and/or gender interactions were present at 15 of these QTL, indicating that such interactions play a significant role in defining the genetic architecture of complex traits such as body weight and obesity. PMID- 18286335 TI - Mapping of genetic modifiers of Nr2e3 rd7/rd7 that suppress retinal degeneration and restore blue cone cells to normal quantity. AB - The retinal degeneration 7 (rd7) mouse, lacking expression of the Nr2e3 gene, exhibits retinal dysplasia and a slow, progressive degeneration due to an abnormal production of blue opsin-expressing cone cells. In this study we evaluated three strains of mice to identify alleles that would slow or ameliorate the retinal degeneration observed in Nr2e3 (rd7/rd7) mice. Our studies reveal that genetic background greatly influences the expression of the Nr2e3 (rd7/rd7) phenotype and that the inbred mouse strains CAST/EiJ, AKR/J, and NOD.NON-H2 (nb1) carry alleles that confer resistance to Nr2e3 (rd7/rd7)-induced retinal degeneration. B6.Cg-Nr2e3 (rd7/rd7) mice were outcrossed to each strain and the F(1) progeny were intercrossed to produce F(2) mice. In each intercross, 20-24% of the total F(2) progeny were homozygous for the Nr2e3 (rd7/rd7) mutation in a mixed genetic background; approximately 28-48% of the Nr2e3 (rd7/rd7) homozygotes were suppressed for the degenerative retina phenotype in a mixed genetic background. The suppressed mice had no retinal spots and normal retinal morphology with a normal complement of blue opsin-expressing cone cells. An initial genome scan revealed a significant association of the suppressed phenotype with loci on chromosomes 8 and 19 with the CAST/EiJ background, two marginal loci on chromosomes 7 and 11 with the AKR/J background, and no significant QTL with the NOD.NON-H2 (nb1) background. We did not observe any significant epistatic effects in this study. Our results suggest that there are several genes that are likely to act in the same or parallel pathway as NR2E3 that can rescue the Nr2e3 (rd7/rd7) phenotype and may serve as potential therapeutic targets. PMID- 18286337 TI - Sacrococcygeal pilonidal sinus treated by a new fascio-cutaneous flap. AB - PURPOSE: This study reported the technical details and preliminary clinical outcomes of a new fascio-cutaneous transposition flap for the surgical treatment of pilonidal sinus. METHODS: Fifty-two patients with pilonidal sinus were surgically treated. During the surgical procedure, an inferiorly pedicled, fascio cutaneous flap with specific geometric characteristics was prepared and transposed. Postoperative pain, complications, duration of hospital stay, and time off worked were assessed. Patients were followed for eighteen months after surgery. RESULTS: The mean hospital stay was 2 days. Drains and sutures were removed after 3 and 12 days, respectively. No flap ischemia, wound dehiscence or major complications were observed. Tension on suture lines and pain after surgery were negligible. Wound infections occurred in 4 patients (7.7 percent) and were managed by removing a few sutures. Seroma was detected in 6 patients (11.5 percent). The mean time patients missed work was 7 days. No recurrence was observed during the follow-up period of 18 months. CONCLUSIONS: With no disease recurrence, minimal complication rate, time off work, and acceptable aesthetic outcome, this fascio-cutaneous transposition flap technique is a safe and effective method for surgically treating pilonidal sinus. PMID- 18286339 TI - Tumor budding as an index to identify high-risk patients with stage II colon cancer. AB - PURPOSE: High-risk patients with Stage II colon cancer may benefit from adjuvant chemotherapy, but they are difficult to identify. We assessed the value of tumor budding, defined as small clusters of undifferentiated cancer cells at invasive margins, as a predictor of outcomes in patients with Stage II colon cancer. METHODS: We studied a total of 200 patients with Stage II colon cancer who underwent curative surgery. With hematoxylin and eosin-stained specimens, the degree of tumor budding was classified as low-grade or high-grade. The survival rate of patients who had Stage II disease with low-grade or high-grade tumor budding was compared with that of 226 patients who had Stage III colon cancer. RESULTS: Univariate analysis revealed that serosal surface involvement (P = 0.04) and tumor budding (P < 0.001) were significantly related to survival. Cumulative five- and ten-year survival rates differed significantly between patients with low-grade tumor budding (93.9 and 90.6 percent, respectively) and those with high grade (73.9 and 67.8 percent, respectively). Survival rates did not differ significantly between patients with Stage II disease who had high-grade tumor budding and patients with Stage III disease. Cox's regression analysis demonstrated that tumor budding (hazard ratio, 4.89; P < 0.001) and serosal surface involvement (hazard ratio, 2.561; P = 0.023) were independent prognostic factors. Liver (P < 0.001) and peritoneal (P = 0.003) metastases were more frequent in the patients with high-grade tumor budding than in those with low grade. CONCLUSIONS: Tumor budding is useful for prognosis and identifying patients with Stage II colon cancer who have a high risk of disease recurrence after curative surgery. PMID- 18286340 TI - Responses of Helicoverpa armigera to tomato plants previously infected by ToMV or damaged by H. armigera. AB - We report the comparative inducing effects of a phytopathogen and a herbivorous arthropod on the performance of an herbivore. Tomato, Lycopersicon esculentum Mill., was used as the test plant, and tomato mosaic virus (ToMV) and corn earworm, Helicoverpa armigera Hubner, were used as the phytopathogen and herbivore, respectively. There were decreases in the efficiency of conversion of ingested food and efficiency of conversion of digested food when H. armigera was reared on tomato plants that had been previously inoculated with ToMV. However, virus inoculation did not affect feeding or oviposition preferences by H. armigera. In contrast, approximate digestibility, total consumption, relative growth rate, and relative consumption rate were lower for fourth-instar H. armigera that fed on plants previously damaged by the same herbivore. Feeding and oviposition were both deterred for H. armigera that fed on previously damaged plants. The duration of development of H. armigera was also prolonged under this treatment. Infection by ToMV and feeding damage by H. armigera increased the host plant's peroxidase and polyphenol oxidase activity, respectively, suggesting that the performance of H. armigera may be affected by the induced phytochemistry of the host plant. Overall, this study indicated that, in general, insect damage has a stronger effect than ToMV infection on plant chemistry and, subsequently, on the performance of H. armigera. PMID- 18286341 TI - Live, attenuated varicella zoster vaccination of an immunocompromised patient. AB - A vaccine for the prevention of herpes zoster outbreaks in adults over the age of 60 years has recently been approved. A 76-year-old white female with a history of recurrent left axillary breast cancer undergoing chemotherapy was given a Zostavax injection by her primary care physician. Eight days later, the patient developed a rash. Given the recent administration of live, attenuated varicella zoster virus (VZV), a diagnosis of disseminated cutaneous herpes zoster was made. The patient was treated successfully with a course of famciclovir for 10 days and cephalexin for 7 days for a secondary bacterial infection. A review of the medical literature disclosed no reports of Zostavax given to adult cancer patients immunocompromised by systemic chemotherapy. Therefore, we believe this report is the first to describe the consequences of Zostavax administration to such a host. Clinicians should take care to review contraindications and precautions prior to administering the Zostavax vaccine. PMID- 18286342 TI - Desiderata or dogma: what the evidence reveals about physician attire. AB - INTRODUCTION: Physician-patient interactions are complex and depend on multiple factors including common cultural definitions and evolving social norms. The once purely philosophical debate over what constitutes appropriate physician attire can benefit from a growing evidence base in the literature. DISCUSSION: Although this literature is commonly regarded as supporting traditional attire, the data actually represent a more balanced distribution of opinions held by patients and by physicians. Perhaps interpretations favoring a conservative approach are expected given the history and tradition of the physician-patient relationship. CONCLUSION: Nevertheless, in the age of evidence-based medicine, it is difficult to argue against scrutiny of the available literature. Evidence that challenges the importance of traditional physician attire is reviewed. PMID- 18286344 TI - Variations in practice guideline adherence for abnormal cervical cytology in a county healthcare system. AB - BACKGROUND: Reduction in cervical cancer incidence and mortality is not only dependant on promoting cervical cancer screening but also on providing appropriate follow-up and treatment of abnormal cervical cytology. OBJECTIVES: The objective of this study was to determine variations in guideline adherence for women requiring abnormal cervical cytology follow-up. SUBJECTS: Subjects of the study are women 18 years or older with an abnormal Pap test in 2000 within a large county healthcare system (n = 8,571). MEASUREMENTS: Guideline adherence was determined by the presence or absence of the appropriate follow-up procedure within an acceptable time interval for each degree of cytological abnormality. Patients with no follow-up studies were deemed to be lost to follow-up. RESULTS: Of study subjects, 18.5% were lost to follow-up care. Of the remaining 6,987 women, 60.3% received optimal care, 9.4% received suboptimal care, and 30.3% received poor care. Follow-up rates were higher for patients with higher degree of cytological abnormality (OR, 1.29, 95% CI, 1.17-1.42), older patients (OR, 1.03, 95% CI, 1.02-1.030) and those receiving the index Pap test at a larger healthcare facility (OR, 1.13; 95% CI, 1.01-1.27). Receiving optimal care was positively correlated with higher degree of cytological abnormality (p < .0001) and larger facility size (p = .002). Regional variations in care demonstrated the largest cluster having the lowest lost to follow-up rate and the most optimal care. CONCLUSIONS: A significant number of women with abnormal cervical cytology are receiving less than optimal care. Further studies are required to determine the specific healthcare delivery practices that need to be targeted to improve guideline adherence for follow-up of abnormal cytology. PMID- 18286345 TI - Depressive symptoms, bone loss, and fractures in postmenopausal women. AB - BACKGROUND: Osteoporosis and depression may be associated through common physiologic systems or risk factors. OBJECTIVE: To assess the associations between depressive symptoms (Burnam's scale) or antidepressant use and bone outcomes. DESIGN: Prospective cohort study. PARTICIPANTS: A total of 93,676 postmenopausal women (50 to 79 years old) enrolled in the Women's Health Initiative Observational Study. MEASUREMENTS: Self-reported fractures (n = 14,982) (hip [adjudicated], spine, wrist, and "other"). Analyses included 82,410 women with complete information followed on average for 7.4 years. Bone mineral density (BMD) of the hip (n = 4539), spine (n = 4417), and whole body (n = 4502) was measured at baseline and 3 years in women enrolled at 3 densitometry study sites. RESULTS: Overall, there were no statistically significant associations between depressive symptoms or antidepressant therapy and 3-year change in BMD. In a subset of women not using antidepressants, there was a significant difference in whole-body BMD change between women with and without depressive symptoms (P = .05). Depressive symptoms (hazard ratio [HR] 1.08; 95% CI = 1.02 to 1.14) and antidepressant therapy (HR = 1.22; CI = 1.15 to 1.30) independently increased risk of any fracture, the majority of which occurred at "other" anatomic sites. Antidepressant therapy increased the risk of spine fracture (HR = 1.36; CI = 1.14 to 1.63). No associations were observed between depressive symptoms or antidepressant therapy and hip or wrist fracture. CONCLUSION: In this study of postmenopausal women, average age 64, we observed minimal association between depressive symptoms and 3-year changes in either BMD or fracture risk. Antidepressant use was not associated with changes in BMD, but was associated with increased risk of fractures at the spine and "other " anatomic sites. PMID- 18286346 TI - Support group meeting attendance is associated with better weight loss. AB - BACKGROUND: Support group meetings (SGM) are assumed to be an integral part of success after bariatric surgery. This investigation studies the effect of SGM on weight loss as well as factors associated with attendance of SGM. It is our hypothesis that patients who attend SGM (ASGM) lose more weight than those patients who do not attend SGM (NASGM). METHODS: Postoperative bariatric patients completed a questionnaire regarding their opinions of SGM. Change in body mass index (BMI) was computed for each patient. The patients were then divided into two groups: ASGM and NASGM for data comparison. RESULTS: There were 46 patients in the investigation. Patients in the NASGM group tended to feel that SGM are not needed after bariatric surgery compared to the ASGM group (5.29 vs. 7.06; p = 0.07). Patients in the NASGM group tended to feel that they would lose the same amount of weight with or without attending SGM compared to the ASGM group (5.67 vs. 7.38; p = 0.07). There were no differences in distance to clinic nor in time to clinic between both groups. Gastric bypass patients in the ASGM group had a statistically significantly higher percent decrease in BMI than the patients in the NASGM group (42% vs. 32%; p < 0.03). CONCLUSION: Patients in the ASGM group lose more weight than patients in the NASGM group. The importance of attending SGM should be incorporated in preoperative patient counseling and encouraged during postoperative follow-up visits. PMID- 18286347 TI - Endoscopic ultrasound-guided gastric botulinum toxin injections in obese subjects: a pilot study. AB - BACKGROUND: Gastric injections of botulinum toxin A (BTA) may induce changes in gastric emptying and body weight, but results vary. BTA dose and depth of injection may affect efficacy. This study assessed changes in gastric emptying, satiation, symptoms, and body weight after endoscopic ultrasound (EUS)-guided injection of 100 or 300 U BTA into gastric antral muscularis propria of obese subjects. METHODS: Open label study of ten healthy, obese adults (age = 29-49 years, body mass index = 31-54 kg/m(2)) who received 100 U (n = 4) or 300 U (n = 6) BTA and were followed for 16 weeks. Measures included gastric emptying of solids (by scintigraphy), satiation (by maximum tolerated volume [MTV] during nutrient drink test), gastrointestinal symptoms (by the Gastrointestinal Symptom Rating Scale), caloric intake (by food frequency questionnaire), and body weight. RESULTS: For the entire cohort, MTV decreased from 1,380 cc (range: 474-2,014) at baseline to 620 cc (range: 256-1,180) 2 weeks after BTA injection; decreases were statistically significant in the subjects receiving 300 U BTA (p = 0.03). Average body weight loss was 4.9 (+/-6.3) kg after 16 weeks. Gastric emptying T(1/2) was prolonged in the 300 U BTA group, but not significantly different from baseline (p = 0.17). BTA injections were well tolerated without significant adverse effects. CONCLUSION: EUS-guided injection of BTA into gastric muscularis propria can be performed safely with minimal adverse effects. A dose of 300 U BTA significantly enhances satiation, is associated with weight loss, and may slow gastric emptying. Further study of higher dose BTA in obese subjects is warranted. PMID- 18286348 TI - High prevalence of small intestinal bacterial overgrowth in patients with morbid obesity: a contributor to severe hepatic steatosis. AB - BACKGROUND: With the increasing prevalence of obesity, non-alcoholic fatty liver disease (NAFLD) has become a major cause of liver diseases. Small intestinal bacterial overgrowth (SIBO) could be related to NAFLD. Our aim was to determine the prevalence of SIBO and its relationship with liver lesions in morbidly obese patients. METHODS: A glucose hydrogen (H(2)) breath test (positive if fasting breath H(2) concentration > 20 ppm and/or an increase of > 10 ppm over baseline within the first 2 h) was performed in obese patients referred for bariatric surgery (body mass index [BMI] > 40 kg/m(2) or > 35 in association with comorbidities) and in healthy non-obese subjects. In obese patients, a surgical liver biopsy was performed. RESULTS: One hundred and forty-six patients (129 women, age [mean+/-SE]: 40.7 +/- 11.4 years) were prospectively included in the study. The mean BMI was 46.1+/-6.4 kg/m(2). A liver biopsy was available in 137 patients and a breath test in 136. The frequency of positive breath tests was higher in obese patients (24/136, 17.1%) than in healthy subjects (1/40, 2.5%; P=0.031). In the univariate analysis, SIBO was not associated with clinical variables, but tended to be associated with more frequent severe hepatic steatosis (26.3 vs. 10.3%, P=0.127), whereas the frequency of sinusoidal or portal fibrosis, lobular necrosis and non-alcoholic steatohepatitis (NASH) were not different. In the multivariate analysis, SIBO (P=0.005) and the presence of a metabolic syndrome (P=0.006) were independent factors of severe hepatic steatosis. CONCLUSION: In morbidly obese patients, bacterial overgrowth prevalence is higher than in healthy subjects and is associated with severe hepatic steatosis. PMID- 18286350 TI - Mesh fixation using staples in Lichtenstein's inguinal hernioplasty: fewer complications and fewer recurrences. AB - BACKGROUND: Fixation of the mesh in Lichtenstein's inguinal hernioplasty is traditionally performed with polypropylene sutures. A modification of this technique uses staples for securing of the mesh. METHOD: A retrospective comparative study of 149 elective repairs of a primary inguinal hernia was performed: a control group of 67 patients undergoing mesh fixation using sutures and a study group of 82 patients undergoing staple fixation. Operating time, recurrence, postoperative pain, complications and costs were studied. RESULTS: Seven recurrences (11%) occurred in the polypropylene group as compared to one recurrence (1%) in the staple group (P < 0.01). There was a trend of fewer complications in the staple group. Operative time and long-term postoperative pain did not differ significantly between the two groups. The costs per surgery for mesh fixation and skin closure were euro 11.13 for the suture group and euro 24.35 for the staple group. CONCLUSION: Staple fixation of the mesh in Lichtenstein's inguinal hernioplasty can be considered equal to traditional fixation with sutures with regard to operating time and postoperative pain. However, staple fixation seems to show fewer recurrences and fewer complications. PMID- 18286349 TI - The "Birmingham stitch"--avoiding slippage in laparoscopic gastric banding. AB - BACKGROUND: Slippage rates of 1.4-24 % are frequently quoted after adjustable gastric banding. This complication can be extremely serious and has contributed to many units offering more invasive interventions in the surgical management of morbid obesity. We present results of the first 1,140 Laparoscopic Bands performed in our unit. METHODS: Between April 2003 and June 2007, 1140 consecutive patients, mean weight 121.5 kg (range 73-268 kg), mean body mass index (BMI) 44.3 kg/m(2) (range 35-88) underwent laparoscopic adjustable gastric banding (LAGB). An identical surgical technique of one gastropexy suture in addition to the two routine gastro-gastro tunnel sutures was used in all cases. Fluoroscopy-guided adjustments were performed at 3 and 6 months and fluoroscopic evaluations were performed later if clinically indicated. RESULTS: There was no mortality and only one major septic complication of gastric perforation 1 week postoperatively which was managed conservatively. The mean stay was 1.02 days (range 0-30 days). Excess percent BMI loss in these patients at 3, 6, 12, 18, 24, 30, and 36 months were 25.4%, 34.7%, 38.3%, 41.1%, 43.7%, 44.4%, and 58.9%, respectively. Slippage with urgent readmission occurred in one patient (0.08%) at 5 months. Two partial slippages were noticed at 12 and 18 months, respectively. One patient had the band removed and the other was treated by band deflation and repositioning 6 months later. CONCLUSION: These results demonstrate that in our unit, laparoscopic gastric band insertion is successful in producing weight loss and at the same time has a very low slippage and pouch dilatation rate. This difference is most probably secondary to operative technique. PMID- 18286351 TI - Clinical assessment of low-dose steroid therapy for patients with IgA nephropathy: a prospective study in a single center. AB - BACKGROUND/AIM: No accepted therapy has been established for progressive IgA nephropathy (IgAN). The purpose of the present study was to assess low-dose steroid therapy in the treatment of patients with IgAN. METHODS: A prospective trial of low-dose steroid therapy was performed in patients with IgAN with mild histological activities. Twenty-four patients in the steroid group and 24 patients in the control group were included in this study. The initial dose of prednisolone was 0.4 mg/kgBW/day (20-30 mg/day), gradually tapered to 5-10 mg/day over 24 months. The patients with mild active inflammatory lesions were treated with prednisolone. The patients assigned to the control group were treated with dipyridamole or zilazep hydrochloride in a dose of 150 or 300 mg/day. RESULTS: In all of the patients studied, serum creatinine levels did not significantly change over 24 months. However, daily proteinuria significantly reduced after 24 months of steroid therapy (0.97 +/- 0.75 vs. 0.31 +/- 0.51 g/day, P = 0.0012), even if did not change after 24 months of anti-platelet drugs (0.89 +/- 0.49 vs. 0.68 +/- 0.69 g/day, P = 0.2289), respectively. In addition, the grade of hematuria significantly reduced after 24 months of steroid therapy (35.6 +/- 36.3 RBC/HPF vs. 13.7 +/- 28.4 RBC/HPF, P = 0.0249) and 24 months of anti-platelet drugs (30.1 +/- 37.1 RBC/HPF vs. 12.4 +/- 20.3 RBC/HPF, P = 0.0465), respectively. Systolic and diastolic blood pressures did not significantly change during treatment with steroid or anti-platelet drugs. Vascular changes (0.63 +/- 0.73) in the steroid group were lower than those (1.08 +/- 0.88) in the control group (P = 0.008). CONCLUSION: Our data suggested that low-dose steroid therapy for IgAN patients with mild inflammatory lesions could reduce the amount of urinary protein excretion and prevent deterioration of renal function, provided the histological findings in the renal biopsies showed mild vascular lesions. PMID- 18286353 TI - Cavitary lung lesion in a patient with systemic lupus erythematosus: an unusual manifestation of cytomegalovirus pneumonitis. AB - We report a 35-year-old female patient with systemic lupus erythematosus (SLE). She was admitted due to deterioration of lupus nephritis and received treatment with a high dose of steroid and cyclosporine. Approximately 1 month after admission, the patient was also treated for cytomegalovirus (CMV) infection because she was found to have CMV antigenemia. Although a cavitary lesion was shown by chest computed tomography (CT), its cause could not be clarified by blood examination, smears or cultures, or by bronchoscopy. We considered that this lesion may have been caused by CMV pneumonitis because it was resolved during the treatment for CMV infection. It is known that CMV causes opportunistic infections in patients with collagen vascular diseases (CVD) who are receiving immunosuppressive therapy. However, it is extremely rare for a cavitary lesion to be formed as a result of CMV pneumonitis. Here we describe the details of this interesting case. PMID- 18286352 TI - Osteonecrosis and monoarticular rheumatoid arthritis treated with intra-articular adalimumab. AB - We report a 54-year-old patient with RA presented with osteonecrosis (ON) and monoarthritis of left knee remitting in the last 2 years. Monoarthritis was resistant to disease-modifying antirheumatic drugs (DMARDs) and intra-articular corticosteroids. Intra-articular administration of adalimumab provided a good clinical and radiographic response. Synovitis resolved and osteonecrosis disappeared almost totally. PMID- 18286354 TI - Atherothrombotic brain infarction in a patient with pachydermoperiostosis. AB - A 58-year-old man was admitted due to mild right-sided hemiparesis and a speech disturbance. We diagnosed an atherosclerotic brain infarction of the left cerebral hemisphere. The patient had severe clubbing of his fingers and toes bilaterally. Based on the history, clinical features, and X-ray examinations, he was diagnosed as having pachydermoperiostosis. Although general complications have been reported in previous literature, this is the first report of a patient with pachydermoperiostosis who had a cerebrovascular event. PMID- 18286355 TI - Therapeutic efficacy of intravenous cyclophosphamide concomitant with moderate- to high-dose prednisolone in two patients with fasciitis panniculitis syndrome. AB - Fasciitis panniculitis syndrome (FPS) has been proposed as a new category of 'fasciitis' and includes the well-established eosinophilic fasciitis (EF). Unlike EF, FPS exhibits inconsistent eosinophilia and/or eosinophilic infiltration of the lesions. Principal histological FPS findings include dermal thickening, inflammation and thickening of the subcutaneous fat tissue, fibrous thickening of the fascia and inflammation of the adjacent muscle. FPS is commonly resistant to corticosteroids, and cimetidine is effective in approximately 80% of FPS patients. A new therapy for FPS is required for cases refractory to treatment or intolerant to cimetidine because of adverse drug reaction. In this report, two FPS patients were resistant to corticosteroids. Both received intravenous cyclophosphamide (IVCY) concomitant with moderate- to high-dose prednisolone (PSL), and this effectively treated the induration of the FPS lesions. Patient 1 was a 50-year-old woman who had been diagnosed with fasciitis following en bloc muscle biopsy of the thigh. She had been treated with high-dose PSL for 6 years, but the fasciitis was refractory. Induration of the neck, thorax and thighs resulted in impaired neck rotation, restrictive respiratory failure and impaired walking. A diagnosis of FPS was made by re-assessing the en bloc muscle biopsy. Although PSL (40 mg/day) for 18 days was ineffective, the addition of IVCY (400 mg) dramatically improved the disease manifestations. Patient 2 was a 68-year-old man who was diagnosed with fasciitis based on en bloc muscle biopsy of the left foot. He had been treated with PSL for 16 years, but the fasciitis was refractory. He exhibited lower limb induration and a refractory skin ulcer of the left foot. A diagnosis of FPS was made by re-assessing the en bloc muscle biopsy. Although PSL (40 mg/day) for 2 weeks was ineffective, the addition of IVCY (450 mg) improved both the lower limb induration and the skin ulcer. FPS may cause both entrapment vasculopathy of subcutis and perivasculitis of the subcutaneous fat tissue such that the skin ulcer might be closely related with the ischemic mechanism triggered by FPS. According to the clinical courses of our cases, IVCY combined with moderate- to high-dose PSL may be a new therapeutic choice for corticosteroid-resistant FPS patients. PMID- 18286356 TI - Carnosol, a dietary diterpene, displays growth inhibitory effects in human prostate cancer PC3 cells leading to G2-phase cell cycle arrest and targets the 5'-AMP-activated protein kinase (AMPK) pathway. AB - PURPOSE: This study examines the anti-cancer effect of carnosol in human prostate cancer PC3 cells and its role in modulating multiple signaling pathways associated with carcinogenesis. METHODS: PC3 cells were treated with carnosol and were evaluated using a flow cytometry, a protein array and Western blot analysis to identify signaling pathways targeted by carnosol. RESULTS: Using an MTT assay we found that carnosol (10-70 microM) decreases cell viability in a time and dose dependent manner. Further analysis using flow cytometry as well as biochemical analysis identified G2-phase cell cycle arrest. To establish a more precise mechanism, we performed a protein array that evaluated 638 proteins involved in cell signaling pathways. The protein array identified 5'-AMP-activated protein kinase (AMPK), a serine/threonine protein kinase involved in the regulation of cellular energy balance as a potential target. Further downstream effects consistent with cancer inhibition included the modulation of the mTOR/HSP70S6k/4E BP1 pathway. Additionally, we found that carnosol targeted the PI3K/Akt pathway in a dose dependent manner. CONCLUSIONS: These results suggest that carnosol targets multiple signaling pathways that include the AMPK pathway. The ability of carnosol to inhibit prostate cancer in vitro suggests carnosol may be a novel agent for the management of PCa. PMID- 18286357 TI - Formation of stable submicron protein particles by thin film freezing. AB - PURPOSE: Highly stable, submicron lactate dehydrogenase (LDH) and lysozyme particles may be produced by thin film freezing (TFF) of aqueous solutions followed by lyophilization. METHODS: The LDH activity was determined by measuring the decrease in absorbance of NADH over time for the reaction of pyruvate to lactate. For lysozyme the particle morphology was determined by scanning electron microscopy (SEM) and compared with the specific surface area (BET) and the particle size, as measured by laser light scattering, RESULTS: Protein particles with an average diameter of 300 nm and 100% enzyme activity upon reconstitution (for LDH) were formed by TFF. Droplets of protein solutions, 3.6 mm in diameter, spread upon impact with 223 and 133 K metal surfaces to form cylindrical disks with thicknesses of 200-300 microm. Calculated cooling rates of the disks of 10(2) K/s were confirmed experimentally with infrared measurements. CONCLUSIONS: The cooling rates of 10(2) K/s, intermediate to those in lyophilization (1 K/min) and spray freeze-drying (SFD) (10(6) K/s), were sufficiently fast to produce sub micron protein particles with surface areas of 31-73 m2/g, an order of magnitude higher than in lyophilization. In addition, the low surface area/volume ratio (32 45 cm(-1)) of the gas-liquid interface led to minimal protein adsorption and denaturation relative to SFD. PMID- 18286358 TI - Content and concurrent validity of the motivation for change questionnaire. AB - INTRODUCTION: Musculoskeletal disorders (MSD) are nowadays seen within a biopsychosocial framework, including salutogenic factors, motivation factors, and coping ability. Such a framework recognizes the importance of motivational factors in health promotion and in rehabilitation. The Motivation for Change Questionnaire (MCQ) has been developed to measure the strength of individuals' motivation for change in life, MCQ part 1, and work situation, MCQ part 2. The purpose of the study was to test the content and concurrent validity of the MCQ on patients with prolonged musculoskeletal disorders referred to interdisciplinary rehabilitation as a basis for use in medical and occupational rehabilitation. METHODS: Content validity was studied among an expert group of 20 rehabilitation professionals at a rehabilitation centre, and with 10 individuals suffering from prolonged MSD in the south of Sweden. The experts evaluated the clinical relevance of each question in MCQ. Concurrent validity was studied on 58 patients with prolonged MSD at an interdisciplinary rehabilitation centre in the south of Sweden. They answered MCQ, QPS Nordic questionnaire, KASAM and the Action theory questionnaire. Spearman's rank correlation coefficient was used in the analyses. RESULTS: The MCQ covered and measured areas of relevance according to content validity. No floor effects in any of the subscales of MCQ part 1 were seen. In MCQ part 2, floor effects were seen in two sub indexes. As for concurrent validity subscales of MCQ correlated significantly with QPS Nordic questionnaire and KASAM. CONCLUSIONS: Findings so far indicate the instrument to be valid for use within the present patient group. The questionnaire can be used to identify patient's motivating factors for change in life and work, as a basis for motivational work within rehabilitation. PMID- 18286359 TI - Significance of the conserved Tyr352 and Asp380 residues in the catalytic activity of Bacillus stearothermophilus aminopeptidase II as evaluated by site directed mutagenesis. AB - The importance of the conserved Tyr352 and Asp380 residues of Bacillus stearothermophilus aminopeptidase II (AP-II) was investigated by site-directed mutagenesis. The wild-type and mutant enzymes were expressed in recombinant Escherichia coli M15 cells and the 45-kD proteins were purified from the cell free extracts by Ni(2+)-NTA resin. The specific activity for Tyr352 and Asp380 replacements was decreased by more than 3.5-fold. Detailed analysis of the kinetic consequences in the mutant proteins revealed that the K (m) values were increased 1.9- to 2.6-fold with respect to wild-type enzyme. Catalytic efficiencies (k (cat)/K (m)) of mutant proteins were between 3.5- and 31-fold lower than the corresponding value of the wild-type enzyme. Tryptophan emission fluorescence and circular dichroism spectra were nearly identical for wild-type and mutant enzymes. These results indicate that residues Tyr352 and Asp380 are essential for the proper function of AP-II. PMID- 18286360 TI - Genetics in health practice and education special issue. PMID- 18286361 TI - Determinants of perceived morbidity and use of health services by children less than 15 years old in rural Bangladesh. AB - This study examined the association of a number of social and economic and other factors with perceived morbidity and use of health services by children in rural Bangladesh, using the data of a health and socioeconomic survey conducted in Matlab, Bangladesh in 1996. One of the factors of interest was women's social position measured with indicators such as their education, domestic autonomy, social networks and social prestige. Other factors of interest were economic in nature and included the availability of high-quality primary health care (PHC) facilities in one part of the study area. A total of 52% of the 3,793 children below 15 had an episode of an acute illness in the month preceding the interview. The medical care sought for acute illnesses was grouped into four categories: medical doctors, paramedics, traditional and untrained village doctors (including drug sellers) and homeopaths. A total of 55% of the children who were sick in the past month consulted any type of health provider. Logistic regression was used to estimate the effects of the various independent variables on the two dependent variables: perceived morbidity of under-15 children and health service use for under-15 sick children. The results revealed that age of the child was the most important factor influencing perceived morbidity while social and economic variables were in general not related to perceived morbidity. Prolonged and severe illnesses and illnesses of young and male children were more likely to be treated by health providers, particularly by physicians. While women's education and social network influenced visits to any health providers socioeconomic indicators influenced visits to physicians. Availability of PHC facilities in one part of the study area also led to more use of modern medical care. The findings highlight that improvement of women's education and of social and economic status in general, in combination with more availability of high-quality PHC will in Bangladesh lead to better health care of children. PMID- 18286362 TI - Epicardial fat from guinea pig: a model to study the paracrine network of interactions between epicardial fat and myocardium? AB - BACKGROUND: Epicardial adipose tissue (Epi) is a fat depot which is closely apposed on the myocardium. Several lines of evidence suggest that it is not only a lipid-storing depot, but also an active endocrine organ which secrets inflammatory cytokines and chemolines as suggested for other types of visceral fat. OBJECTIVES AND METHODS: We selected guinea pigs which have been shown to expand Epi with age, to investigate the expansion and properties of Epi and its impact on cardiac structure and function in detail. RESULTS: The amount of epicardial fat increases rapidly with age and accumulates at the aortic arch. It extends over the anterior surface of both ventricles and along the anterior and posterior branches of the coronary arteries. It also expands within the epicardium. The pattern of cytokines released by Epi is altered with age showing an up- and down-regulation of a variety of the 120 cytokines analyzed. Most prominently changed are IGFBP-4 and TIMP-2, whereas the release of adiponectin is not modified by age. CONCLUSIONS: The amount of Epi is closely correlated to the amount of other types of visceral fat, to insulin resistance and other features of the metabolic syndrome, but also to cardiac hypertrophy and cardiac dysfunction. The data provide evidence that the guinea pig heart is a suitable model to analyze the interactions between Epi, heart vessels and muscle tissue. It allows identifying the influence of nutritional and metabolic alterations on the complexity of the network of locally released mediators for heart structure and function. A deeper understanding of this animal model may be helpful to analyze the interactions between Epi and the myocardium in humans-where the availability of tissue and the possibilities to modify nutritional and metabolic influences on heart are restricted-and the impact of Epi on cardiovascular risk. PMID- 18286363 TI - Use of coercive sexual tactics across 10 years in at-risk young men: developmental patterns and co-occurring problematic dating behaviors. AB - Men's use of two coercive sexual tactics was tracked over 10 years in a sample of at-risk young men (N = 201). Patterns were identified for each tactic. For the tactic using drugs or alcohol to go further sexually, non-coercers (63%) and coercers (37%) were identified. For the tactic of going further sexually after the woman said "no," three patterns were identified-noncoercers (10%), low-level coercers who used the tactic five times or less over 10 years (42%), and high level coercers who used the tactic more than five times over 10 years (48%). The associations between coercive tactics and two dating behaviors-physical aggression toward a partner and risky sexual behaviors-were examined using multilevel linear modeling. For both coercive tactics, main effects and interaction effects with time occurred for physical aggression toward a partner. The most coercive men perpetrated the most physical aggression toward a partner between ages 18 and 22 years, but sexual coercion was unrelated to partner abuse between ages 22 and 27 years. Results suggest men vary in their use of coercive sexual tactics over time and the frequency of coercion varies based on tactic. Preliminary evidence suggests the use of coercive sexual tactics is associated with physical aggression toward a partner but not risky sexual behaviors, though the strength of the association varies over time. PMID- 18286364 TI - A qualitative exploration of the relationship between racism and unsafe sex among Asian Pacific Islander gay men. AB - Although reported cases of HIV/AIDS among gay Asian Pacific Islander (API) American men and API men who have sex with men (MSM) are still relatively low, current research findings indicate that incidences of unsafe sexual activity may be higher for this group than for any other group. Among the explanations offered to explain the levels of increasing unsafe sex among gay API men have been sexual norms found in Asian cultures, the lack of culturally relevant and/or linguistically appropriate intervention material, lack of integration into the mainstream gay community, and internalized homophobia. What are often ignored in these analyses are the contextual norms in which sexual behavior for gay API men occur. In this article, I develop the argument that racism within the gay community leads to socially and contextually prescribed sexual roles for gay API men that may also contribute to the practice of unsafe sex among this group. PMID- 18286365 TI - Family planning among HIV positive and negative prevention of mother to child transmission (PMTCT) clients in a resource poor setting in South Africa. AB - The purpose of this study was to investigate family planning needs, knowledge of HIV transmission and HIV disclosure in a cohort sample that had undergone PMTCT in a resource poor setting. Five public clinics implementing PMTCT from Qaukeni Local Service Area, O.R. Tambo District in the Eastern Cape. The sample at postnatal care consisted of 758 women with known HIV status. From 116 HIV positive women 76.3% and from 642 HIV negative women 85.2% got counseling on safe sex during pregnancy but only 65.8% and 62.3% of the women respectively practiced safe sex during pregnancy, which did not differ by HIV status. Postnatally, almost all women received counseling on family planning, yet use of contraceptives and condoms were low. Among HIV positive women PMTCT knowledge and younger age of the mother were associated with pregnancy desire, and among HIV negative women HIV disclosure to the partner, younger age of the mother and having a lower number of children were associated with pregnancy desire. High pregnancy desires (yet lower than for HIV negative women); low contraceptive and condom use were found among HIV positive women. HIV prevention and family planning must acknowledge the reproductive desires of HIV positive women and men. PMID- 18286367 TI - How culture impacts the dissemination and implementation of innovation: a case study of the Families and Schools Together program (FAST) for preventing violence with immigrant Latino youth. AB - We consider how culture impacts the translation of research into practice, focusing on the culture of the client and the culture of the agency implementing selected programs. We build on lessons learned from a pilot study of an evidence based family-school partnership, Families and Schools Together (FAST), to prevent youth violence with low-income, immigrant Latino families in Southern California. We examine the impact of cultural characteristics on the translation of this innovation into practice at the community level, relying on an interactive systems framework developed recently by Wandersman and colleagues (2008, American Journal of Community Psychology, 41(3-4), in press) discussed in this issue. As we point out, the culture of the client and the culture of the agency can facilitate or impede connections within and across these interactive systems. PMID- 18286366 TI - Reciprocal relationships between parenting behavior and disruptive psychopathology from childhood through adolescence. AB - Theoretical models suggest that child behaviors influence parenting behaviors, and specifically that unpleasant child behaviors coerce parents to discontinue engaging in appropriate discipline. This study examined reciprocal relationships between parenting behaviors (supervision, communication, involvement, timid discipline and harsh punishment) and child disruptive disorder symptoms (ADHD, ODD and CD) in a clinic-referred sample of 177 boys. Annual measures, including structured clinical interviews, were obtained from the beginning of the study (when boys were between the ages of 7 to 12) to age 17. Specific reciprocal influence was observed; only timid discipline predicted worsening behavior, namely ODD symptoms, and ODD symptoms predicted increases in timid discipline. Greater influence from child behaviors to parenting practices was found: ODD also predicted poorer communication and decreased involvement, and CD predicted poorer supervision. ADHD was neither predictive of, nor predicted by, parenting behaviors. The results are specifically supportive of a coercive process between child behaviors and parenting behaviors, and generally suggestive of greater influence of child behaviors on parenting behaviors than of parenting behaviors on child behaviors. PMID- 18286368 TI - Improving youth mentoring interventions through research-based practice. AB - Youth mentoring programs are in the limelight. Over three million young people have a Big Brother, a Big Sister, or a similar adult volunteer involved in their lives-a sixfold increase from just a decade ago-and generous federal funding continues to fuel new initiatives. This expansion speaks volumes about the faith our society places in one-on-one relationships between vulnerable young people and caring adults. But what do we know about the effectiveness of this intervention strategy? A better understanding of the research evidence for youth mentoring, including findings from reviews, evaluations, and meta-analyses, provides a basis for a more informed, practically applicable approach to strengthening youth mentoring interventions. PMID- 18286369 TI - Integrating a portable biofeedback device into clinical practice for patients with anxiety disorders: results of a pilot study. AB - This study examined the effectiveness of a portable Respiratory Sinus Arrhythmia (RSA) biofeedback device as an adjunct to CBT in persons with anxiety disorders and other disorders associated with autonomic dysfunction attending outpatient treatment. Participants were 24 individuals attending outpatient cognitive behavioral treatment for a range of anxiety disorders. Participants were assessed over a 3 week period. Outcomes included measures of anxiety (STAI-Y), sleep disturbances (PSQI), anger (STAEI), and subjective questions about the effectiveness of the device as a treatment adjunct. Significant reductions were found for anxiety and anger and for certain sleep variables (e.g. sleep latency). There was a significant dos-effect in that those who were more compliant had significantly greater reductions in most domains including sleep, anger and trait anxiety. Overall, participants found the device more helpful than other relaxation techniques such as mediation, yoga and unassisted breathing techniques but less helpful than exercise. The most frequently endorsed side effects were dizziness (15%) and sleepiness (55%). These preliminary results suggest that portable RSA biofeedback appears to be a promising treatment adjunct for disorders of autonomic arousal and is easily integrated into treatment. Results support the need for further investigation with more rigorous experimental designs. PMID- 18286370 TI - Comparison among different classification systems regarding the pathological response of preoperative chemotherapy in relation to the long-term outcome. AB - Neoadjuvant chemotherapy (NAC) is increasingly used for operable disease. However there are several pathological response classification systems and the correlation between the pathological response to NAC according to each system and the patient outcome is still under debate. From 1998 to 2006, 370 primary breast cancer patients underwent curative surgical treatment after NAC containing both anthracycline and taxane at the National Cancer Center Hospital. We retrospectively evaluated the clinical and pathological response using the cTMN, Fisher's, Chevailler's, and the Japanese Breast Cancer Society classification systems (JBCS) respectively, and analyzed the correlation between each pathological response and disease free survival (DFS). Ninety-five (26%) patients had tumor recurrence. The five-year DFS according to Fisher's system was pCR, 80% and pINV, 63%. The five-year DFS according to Chevallier's system was Grade 1, 83%, Grade 2, 85%, Grade 3, 62%, and Grade 4, 65%. The five-year DFS according to the JBSC system was Grade 3, 77%, Grade 2, 68%, Grade 1a, 68%, Grade 1b, 58%, and Grade 0, 52%. None of the pathological response systems reached a statistically significant difference. In the classification by the post-treatment number of metastatic axillary lymph nodes, the 5-year DFS was n = 0, 86%; n = 1-3, 64%; n = 4-9, 44%; and n > 10 positive: 25% (P < .0001). In pathologically node negative patients, there were no significant differences in the DFS among all the classification systems. All three classifications analyzed were considered inadequate as the prognostic marker of the long-term outcome after NAC and further studies are warranted to optimize the prediction. PMID- 18286371 TI - High doses of atrazine do not disrupt activity and expression of aromatase in female gonads of juvenile goldfish (Carassius auratus L.). AB - Juveniles female goldfish were exposed to atrazine (2-chloro-4-ethylamino-6 isopropylamino-s-triazine) at high doses, 100 and 1000 microg l(-1) during 56 days in order to evaluate the potential action of the herbicide as an endocrine disruptor. Plasma concentration of estradiol (E2) and 11-ketotestosterone (11-KT) as well as activity and expression of aromatase in the gonads were evaluated. These parameters were completed with morphological measures such as gonadosomatic index (GSI) and histological analyses of gonads. Morphological parameters at both 100 and 1000 microg l(-1) did not show any significant differences with the control groups. Correlated to the pathway hypothesized, no time-, dose-related effects were detected on the aromatase activity and the expression in the gonads of juvenile female goldfish. The same conclusion was attributed regarding the circulating E2 where no perceptible variation was detected. Nevertheless, a hormonal imbalance was detected for plasma concentration of the sex steroid 11-KT of fish exposed to 1000 microg l(-1) after 56 days exposure. In these particular experimental conditions, we failed to demonstrate an effect of atrazine through the induction of aromatase and hormonal imbalance associated. PMID- 18286372 TI - Cost-effectiveness analysis of the Not On Tobacco program for adolescent smoking cessation. AB - Public health researchers and practitioners emphasize the need for effective, adoptable, and available youth smoking cessation interventions. Scarce resources demand that such interventions also be cost effective. This study describes a cost-effectiveness analysis (CEA) of the American Lung Association's Not On Tobacco (N-O-T) national and international teen smoking cessation program. N-O-T has been rigorously evaluated as an effective and adoptable program, and was recently found to be the most frequently-used teen smoking cessation program in the nation. N-O-T studies show intent-to-treat quit rates between 15% and 19%, among the highest reported in the literature. The current CEA resulted from a 2 year state-wide demonstration study in Florida, comparing the effectiveness of N O-T with a 20-min brief intervention (BI). The CEA utilized a Markov transition model of decision analysis to explain stage progression of smoking cessation among participants from the age of 17 to 25 years. The Markov simulation predicted that out of a cohort of 100 N-O-T students, 10 will quit smoking and remain smoke-free at the age of 25 years and 14 will reduce smoking, resulting in 102.22 life years saved and a total of 20.11 years discounted life years (DLY) saved. Among BI youth, six will quit smoking and nine will reduce, indicating 64.31 life years saved and a total 12.65 DLY saved. The incremental DLY saved is 7.46 years. Results indicate that N-O-T is a very cost-effective option school based smoking cessation, as cost effective as school-based primary tobacco prevention, and potentially more cost effective than adult tobacco use cessation. PMID- 18286374 TI - Endoscopic pituitary surgery. AB - Pituitary surgery is a continuous evolving speciality of the neurosurgeons' armamentarium, which requires precise anatomical knowledge, technical skills and integrated appreciation of the pituitary pathophysiology. What we consider "pure" endoscopic transsphenoidal surgery is a procedure performed through the nose and the sphenoid bone, with the endoscope alone throughout the whole approach to visualize the surgical target area and without the use of any transsphenoidal retractor. It offers some advantages due to the endoscope itself: a superior close-up view of the relevant anatomy and an enlarged working angle are provided with an increased panoramic vision inside the surgical area. Concerning results in terms of mass removal, relief of clinical symptoms, cure of the underlying disease and complication rate, they are, at least, similar to those reported in the major microsurgical series, but patient compliance is by far better. Furthermore transsphenoidal endoscopy brings advantages to the patient (less nasal traumatism, no nasal packing, less post-op pain and usually quick recovery), to the surgeon (wider and closer view of the surgical target area, increase of the scientific activity as from the peer-reviewed literature on the topic in the last 10 years, smoothing of interdisciplinary cooperation), to the institution (shorter post-op hospital stay, increase of the case load). Besides, further progress and technological advance are expected from the close cooperation between different technologies and industries. Continuing works in such field of "minimalism" will offer further possibilities to provide the surgeon with even more effectiveness and safety, and, on the other hand, the patient with improvement of results. PMID- 18286373 TI - Radiotherapy of nonfunctioning and gonadotroph adenomas. AB - Transsphenoidal surgery is the treatment of choice for NFPA but is seldom curative. The management of patients in whom residual tumor is detected after surgery is not clear-cut. Radiation therapy is effective in controlling tumor mass in the majority of patients, but is associated with long term complications that call for restriction of its use to patients at high risk for tumor growth. New radiation techniques may prove to be safer while retaining the effectiveness of conventional radiotherapy, however longer follow-up is necessary to confirm this assumption. For now, it appears to be safe to withhold radiation and carefully follow patients with small tumor remnants, whereas large remnants from invasive tumors should be considered for radiotherapy. Nevertheless, there are no prospective controlled studies that support this empirical approach. PMID- 18286376 TI - Characterization of the recombinant Rieske [2Fe-2S] proteins HcaC and YeaW from E. coli. AB - Three genes within the genome of E. coli K12 are predicted to encode proteins containing the typical Rieske iron-sulfur cluster-binding motifs. Two of these, hcaC and yeaW, were overexpressed in E. coli BL21 and Tuner (DE3) pLacI. The recombinant proteins were purified and analyzed by UV/Vis- and EPR-spectroscopy. HcaC and YeaW display the typical redox-dependent UV/Vis-spectra of iron-sulfur proteins. The EPR spectrum of reduced HcaC shows characteristic g-values of a Rieske cluster whereas the g-values for YeaW are close to the upper limit for this type of iron-sulfur cluster. Both iron-sulfur clusters could be reduced by dithionite, but not by ascorbate, confirming their classification as low potential Rieske proteins as derived from the amino acid sequences. A phylogenetic analysis of the two proteins reveals that HcaC clearly segregates with the Rieske ferredoxins of class IIB oxygenases whereas the classification of YeaW remains doubtful. PMID- 18286375 TI - Development and validation of short versions of the internal mental distress and behavior complexity scales in the Global Appraisal of Individual Needs (GAIN). AB - Co-occurring mental distress and behavior problems are the norm in substance abuse treatment but are often poorly assessed due to resource constraints. This paper describes the development and validation of scales measuring internalizing mental distress and externalizing behavior problems that are shorter versions of comorbidity scales found in the full Global Appraisal of Individual Needs (GAIN). GAIN data from two treatment outcome studies, one involving adolescents and the other on adults, were used in the creation and testing of the scales. Subsets of items from the full GAIN scales were selected for the short scales through the application of standard psychometric principles. The short comorbidity scales still have moderate to high reliability and are highly correlated with the full scales. Parallel tests of construct validity show no substantial loss when moving from the longer to shorter versions. The short scales maintain good sensitivity and specificity for predicting diagnostic impressions. PMID- 18286377 TI - Microbiological reduction of hexavalent chromium by indigenous chromium-resistant bacteria in sand column experiments. AB - Indigenous bacteria that are resistant to high concentrations of Cr(VI) were isolated from a Cr-contaminated sediment. Sand column experiments were conducted using the isolated bacteria to investigate microbial effects on Cr(VI) reduction in open systems that simulated subsurface conditions. The indigenous Cr-resistant bacteria appeared to reduce Cr(VI) in the column experiments. When 12 mg/L of Cr(VI) was injected into the columns, the removal efficiencies of Cr(VI) by the isolated bacteria were 39.1%, 62.5%, and 63.6% at 24, 48, and 72 h retention times of Cr(VI) solution, respectively. These results imply that the linear velocity of groundwater or pore water should be less than 0.63 cm/h for effective removal of Cr(VI) in subsurface conditions. In comparison, the noninoculated control column did not show a significant variation in dissolved Cr(VI) concentration. The results indicated that reduction of Cr(VI) was occurring in the column due to the activity of the indigenous bacteria. PMID- 18286379 TI - Personal use of hair dyes and risk of bladder cancer: a meta-analysis of epidemiologic data. AB - OBJECTIVE: We compared, updated, and expanded the analyses of two previous meta analyses of personal hair dye exposure and bladder cancer, and briefly discussed the biological plausibility of a systemic hazard to human health from exposure to para-phenylenediamine (PPD), a key chemical in hair dyes. METHODS: The meta analysis included 11 case-control studies and one cohort study. We evaluated heterogeneity across studies and conducted sensitivity and influence analyses. RESULTS: No association was found between any personal use of hair dye and bladder cancer among women (meta-relative risk [mRR] = 1.01, 95% CI: 0.89-1.14), men (mRR = 0.82, 95% CI: 0.60-1.14), or both sexes combined (mRR = 0.97, 95% CI: 0.87-1.08). No statistically significant mRRs were found among the studies that reported data for permanent hair dye use (mRR = 1.06, 95% CI: 0.89-1.27), duration of any hair dye use (mRR = 1.00, 95% CI: 0.85-1.19), duration of permanent hair dye use (mRR = 1.31 95% CI 0.78-2.19), lifetime applications of any hair dye use (mRR = 1.12 (95% CI: 0.72-1.72) or permanent hair dye use (mRR = 1.59, 95% CI 0.69-3.64), or dark color hair dye use (mRR = 0.94, 95% CI: 0.74 1.19). CONCLUSION: The results of this meta-analysis of epidemiologic studies do not indicate a causal association between personal hair dye use and bladder cancer. PMID- 18286378 TI - Matrix metalloproteinases stimulate epithelial-mesenchymal transition during tumor development. AB - Matrix metalloproteinases (MMPs) are a family of more than 28 enzymes that were initially identified on the basis of their ability to cleave most elements of the extracellular matrix (ECM) but have subsequently been found to be upregulated in nearly every tumor type. As digestion of the ECM is essential for tumor invasion and metastasis, MMPs have been studied for their role in these later stages of tumor development. More recently, exposure to these enzymes has been found to impact cellular signaling pathways that stimulate cell growth at early stages of tumor progression. MMPs have also been found to cleave intracellular targets and so inducing mitotic abnormalities and genomic instability. Emerging evidence indicates that tumor-associated MMPs can also stimulate processes associated with epithelial-mesenchymal transition (EMT), a developmental process that is activated in tumor cells during cell invasion and metastasis. Investigations of potential therapeutic MMP inhibitors aimed at blocking the protumorigenic tissue alterations induced by MMPs have been complicated by the side effects associated with nonspecific inhibition of normal physiological processes; recent investigations have shown how delineation of the extracellular targets and intracellular signaling pathways by which MMP action on cancer cells can induce EMT provides insight into novel therapeutic targets. Here, we provide an overview of recent findings of MMP action in tumors and the mechanisms by which MMPs induce both phenotypic and genotypic alterations that facilitate tumor progression. PMID- 18286380 TI - A case-control study of the protective benefit of cervical screening against invasive cervical cancer in NSW women. AB - OBJECTIVE: To examine the effects of different Pap screening patterns in preventing invasive cervical cancer among women in New South Wales, Australia. METHODS: A total of 877 women aged 20-69 years diagnosed with invasive cervical cancer during 2000-2003 were matched with 2,614 controls by month and year of birth. Screening behavior patterns in 4 years preceding the time of cancer diagnosis in the cases were classified into none (no Pap test in the 4 years), 'irregular' (1 of the 4 years with Pap test(s)), and 'regular' (2 or more of the 4 years with a Pap test), and compared with those in the matched non-cases over the same period. Conditional logistic regression modeling was used to estimate the relative risk, approximated by the odds ratio, of invasive cervical cancer for regular and irregular cervical screening compared with no screening in the previous 4 years, before and after controlling for potential confounders including the first recorded Pap test result in the preceding 6-year reference period. RESULTS: Compared with no screening, irregular Pap screening in the 4 years preceding the cancer diagnosis is estimated to reduce the risk of invasive cervical cancer by about 85% (RR = 0.15, 95% CI: 0.120-0.19); regular Pap screening reduces the risk by about 96% (RR = 0.04, 95% CI: 0.03-0.05). After adjusting for the index Pap test result, the relative risks for invasive cervical cancer were 0.19 (95% CI: 0.13-0.27) for irregular screening and 0.07 (95% CI: 0.04-0.10) for regular Pap screening. CONCLUSIONS: Regular and irregular Pap tests among women aged 20-69 years were highly effective in preventing invasive cancer. At-risk women with no Pap test history should be encouraged to undergo a Pap test every 2 years, but any Pap screening over a 4-year period remains highly protective against future invasive cervical cancer. PMID- 18286381 TI - Pesticide residue level in tea ecosystems of Hill and Dooars regions of West Bengal, India. AB - In the present study we quantified the residues of organophosphorus (e.g. ethion and chlorpyrifos), organochlorine (e.g. heptachlor, dicofol, alpha-endosulfan, beta-endosulfan, endosulfan sulfate) and synthetic pyrethroid (e.g. cypermethrin and deltamethrin) pesticides in made tea, fresh tea leaves, soils and water bodies from selected tea gardens in the Dooars and Hill regions of West Bengal, India during April and November, 2006. The organophosphorus (OP) pesticide residues were detected in 100% substrate samples of made tea, fresh tea leaves and soil in the Dooars region. In the Hill region, 20% to 40% of the substrate samples contained residues of organophosphorus (OP) pesticides. The organochlorine (OC) pesticide residues were detected in 33% to 100% of the substrate samples, excluding the water bodies in the Dooars region and 0% to 40% in the Hill region. The estimated mean totals of studied pesticides were higher in fresh tea leaves than in made tea and soils. The synthetic pyrethroid (SP) pesticide residues could not be detected in the soils of both the regions and in the water bodies of the Dooars. Sixteen percent and 20% of the made tea samples exceeded the MRL level of chlorpyrifos in Dooars and Hill regions respectively. The residues of heptachlor exceeded the MRL in 33% (April) and 100% (November) in the Dooars and 40% (April) and 20% (November) in the Hill region. Based on the study it was revealed that the residues of banned items like heptachlor and chlorpyrifos in made tea may pose health hazards to the consumers. PMID- 18286383 TI - Prevalence of BRCA1 and BRCA2 mutations in breast cancer patients from Cuba. AB - The contribution of BRCA1 and BRCA2 to breast cancer incidence in Cuba has not yet been explored. In order to estimate the proportion of breast cancers due to BRCA1 and BRCA2 mutations in Cuba, and to identify possible Cuban founder mutations, we conducted a study of unselected breast cancer patients from Havana, Cuba. We enrolled 336 women with breast cancer from a large public hospital in the city. A family history of cancer was obtained from each patient and a blood sample was processed for DNA analysis. Mutations in BRCA1 and BRCA2 were sought using a combination of techniques, but all mutations were confirmed by direct sequencing. We were able to successfully complete testing on samples from 307 women. Among these, eight mutations were identified (seven in BRCA2 and one in BRCA1) representing 2.6% of the total, including 10% of familial cases and 10% of cases under age forty. One BRCA2 mutation (c.3394C > T) was found in two women, but no clear example of a founder mutation was identified. In summary, BRCA1 and BRCA2 mutations are not uncommon in Cuban women with breast cancer, but the absence of founder mutations precludes the development of a rapid and inexpensive clinical screening test. PMID- 18286382 TI - Evolution-related amino acids play important role in determining regioselectivity of fatty acid desaturase from Pichia pastoris. AB - Omega(3)-fatty acid desaturase and Delta(12)-fatty acid desaturase of Pichia pastoris with distinguishable regioselectivity and high degree of sequence similarity were chosen for regioselectivity research. Chimeras were constructed in which Histidine-rich boxes 1, 2 and the carboxyl terminal region of omega(3) fatty acid desaturase were replaced with corresponding region of Delta(12)-fatty acid desaturase. The replacement was found to result in a change of regioselectivity from omegay to x + 3 by functionally characterizing these chimeric enzymes in Saccharomyces cerevisae strain INVScI. Using site-directed mutagenesis, we further demonstrated that seven conserved amino acids of omega(3) fatty acid desaturase within the first two Histidine-rich regions are responsible for the regioselectivity switch. Therefore, the regioselectivity of fatty acid desaturases may be better understood by investigating the evolutionary relationships of different fatty acid desaturases. PMID- 18286384 TI - Kainic acid down-regulates NOP receptor density and gene expression in human neuroblastoma SH-SY5Y cells. AB - Nociceptin (N/OFQ) is involved in neuronal excitability and in certain types of seizures. Kainate-induced seizures are associated with increased N/OFQ release in the rat thalamus and hippocampus, causing down-regulation of the N/OFQ receptor (NOP). In this study, we used the neuroblastoma SH-SY5Y cell line as a model to investigate the effects of kainate on NOP receptor density and gene expression. Exposure to kainate (10-50 microM) for 3 h did not affect NOP receptor density. In contrast, a NOP Bmax down-regulation was detected in cells exposed to 10 microM kainate for both 6 and 24 h. Moreover, our data show that kainate causes a decrease in NOP mRNA levels after 3, 6, and 24 h, an effect blocked by the AMPA/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). These findings show that kainate is able to affect the NOP system, both at biosynthesis and receptor density levels in SH-SY5Y cells, and that the kainate ionotropic receptor can contribute to the regulation of the NOP receptor. These data are in agreement with data obtained in vivo and provide new evidence concerning the existence of a cross-talk between NOP and kainate receptors, leading to an interplay between glutamate and N/OFQ circuits. PMID- 18286385 TI - ADNP differential nucleus/cytoplasm localization in neurons suggests multiple roles in neuronal differentiation and maintenance. AB - Complete deficiency in activity-dependent neuroprotective protein (ADNP) results in neural tube closure defects and death at gestation day 9 in mice. ADNP deficient embryos exhibit dramatic increases in gene transcripts associated with lipid metabolism coupled to reduction in organogenesis/neurogenesis-related transcripts. In the pluripotent teratocarcinoma cell line P19, ADNP was shown to interact with specific chromatin regions in the neurodifferentiated state, which was associated with binding to the heterochromatin protein 1 alpha. In this study, using P19 cells as a differentiation model, we showed that ADNP expression and cytoplasm/nucleus distribution is unique in neuronal-differentiated cells compared to cardiovascular and nondifferentiated pluripotent cells. ADNP-like immunohistochemical localization to the neuronal cytoplasm and neurites was shown in this study not only in the cellular model but also in the brain cerebral cortex and olfactory bulb. Small hairpin RNA ADNP downregulation was used to further investigate ADNP involvement in p19 neurodifferentiation. An approximately 80% robust reduction in ADNP led to a substantial reduction in embryoid body formation and a significant reduction (approximately 50%) in neurite numbers. These results position ADNP in direct association with neuronal cell differentiation and maturation. PMID- 18286386 TI - The geo-epidemiology of temporal (giant cell) arteritis. AB - Giant cell arteritis (GCA) is the most common vasculopathy in patients over the age of 50. The majority of data on the geo-epidemiology of GCA is derived from Scandinavia, although there is very good documentation and epidemiological descriptions from studies throughout Europe and North America. There remains, however, a paucity of data on the incidence and prevalence of GCA in North American minority populations, as well as from Africa or Asia. The data that does exist suggests that the incidence of GCA is lower in Hispanic, Asian, and African American populations. It is interesting to note that as the population throughout the world continues to age, we anticipate an increased prevalence of disease based upon increases in annual incidence and improved survival. Considerable research is still needed to identify genetic, environmental, and gender-specific factors that influence not only the etiology, but also the natural history of disease. PMID- 18286387 TI - Physical activity and the regulation of neurogenesis in the adult and aging brain. AB - The discovery that exercise regulates adult hippocampal neurogenesis, that is, the production of new neurons in the adult brain, was surprising news and changed quite fundamentally our view on how physical activity affects the brain. The everyday experience that not all athletes are necessarily smarter than more sedentary fellows and the scientific insight that adult hippocampal neurogenesis is actually a process that ranges on a very small scale raised important questions on the relevance of this finding. We propose that the exercise-related regulation of adult hippocampal neurogenesis is a qualitative rather than a quantitative event and that it is a particularly prominent and suggestive example of activity-dependent cellular plasticity. For rodents, the animals, in which most of this research has been done, cognition is almost inseparable from locomotion. Physical activity, especially exerted over longer periods of time, might indicate to the brain an increased chance of experience those situations rich in complexity and novelty that presumably benefit from more new neurons. We thus propose that it is not isolated physical activity that is "good for the brain", but physical activity in the context of cognitive challenges. This would also explain why few new neurons could be beneficial for successful aging. We here review the current stage of the knowledge how this exercise-induced regulation of neurogenesis might work. PMID- 18286388 TI - Physical activity and neuroprotection in amyotrophic lateral sclerosis. AB - Physical exercise exerts a wide range of benefits on an organism's overall health and well-being. Exercise contributes positively toward an individual's healthy weight, muscle strength, immune system, and cardiovascular health. Indeed, exercise has been demonstrated to reduce life-threatening conditions such as high blood pressure, heart disease, obesity, and diabetes. Of particular interest to this review, exercise has also been shown to be neuroprotective in both the central and peripheral nervous systems. Naturally, such findings apply broadly to the study of neurodegenerative disease with numerous reports demonstrating that exercise has beneficial effects on disease progression. One of the most devastating neurodegenerative diseases is amyotrophic lateral sclerosis (ALS), commonly known as Lou Gehrig's disease in the United States, or motor neuron disease in the United Kingdom, resulting from the progressive loss of brain and spinal cord motor neurons. Several human studies show that moderate exercise regimens improve ALS patients' scoring on functionality tests and ameliorate disease symptoms. Other promising recent works using transgenic mouse models of familial ALS have shown markedly slowed disease progression, improved function, and extension of survival in moderately exercised animals. Possible explanations for these findings include the exercise-induced changes in motor neuron morphology, muscle-nerve interaction, glial activation, and altering levels of gene expression of anti-apoptotic proteins and neurotrophic factors in the active tissue. Here we review the current literature on exercise and motor neuron disease, focusing on rodent and human studies to define the proper type, intensity, and duration of exercise necessary to enhance neuron survival as well discuss current mechanistic studies to further define the exercise-mediated pathways of neuroprotection. PMID- 18286390 TI - Growth factors as mediators of exercise actions on the brain. AB - Physical exercise has long been recognized as highly beneficial for brain and body health. The molecular mechanisms responsible for translation of exercise stimuli in the brain have claimed attention due to mounting evidence for the neuroprotective actions of the exercise and its positive effects in preventing both ageing and neurodegenerative disease. These molecular mediators are currently under investigation with new tools able to yield deep insights into the neurobiology of exercise. In the present work we focus on the evidence pertaining to the mediation of exercise effects by insulin-like growth factor 1 (IGF1), as recent reports suggest that this growth factor shows brain area-specific, temporal rank-sensitive, and behavioural task-dependent features in response to exercise. PMID- 18286391 TI - Lack of mitochondrial DNA deletions in lesions of multiple sclerosis. AB - OBJECTIVE: To test if mitochondrial (mt)DNA deletions accumulate in brains of patients with multiple sclerosis (MS). BACKGROUND: Previous studies demonstrated an accumulation of oxidative damage to mtDNA and decreased activity of mitochondrial enzymes in lesions of MS, where activated immune cells produce increased amounts of reactive oxygen species and nitric oxide. The unknown link between oxidative damage and decreased activity of mitochondrial enzymes may be the accumulation of deletions in mtDNA molecules. mtDNA deletions in the brain have been associated with neurodegeneration and aging. METHODS: mtDNA deletions were quantified by using real-time PCR in laser-dissected, COX-positive and COX negative single neuronal and glial cells from frozen postmortem brain tissue specimens including normal appearing gray (NAGM) and white matter (NAWM) regions and chronic active plaques of MS patients, and gray matter (GM) and white matter (WM) regions of age-matched controls. Three patients with advance Alzheimer's and Parkinson's diseases were included as positive controls. The proportion of deleted mtDNA molecules was correlated with pathology and age. RESULTS: We detected no pathology-related accumulation of mtDNA deletions when comparisons were made among NAGM, NAWM, and plaque of MS brains, or between NAGM-GM and NAWM WM of patients and age-matched controls. However, an accumulation of mtDNA deletions was noted in non-neurological controls beyond 60 years of age and in patients with Alzheimer's and Parkinson's diseases. As expected, the rate of mtDNA deletions was higher in COX- than in COX+ cells. CONCLUSION: While aging and neurodegeneration in PD and AD are associated with accumulation of COX- cells and mtDNA deletions, the pathology of MS is not. PMID- 18286389 TI - Neurogenesis and exercise: past and future directions. AB - Research in humans and animals has shown that exercise improves mood and cognition. Physical activity also causes a robust increase in neurogenesis in the dentate gyrus of the hippocampus, a brain area important for learning and memory. The positive correlation between running and neurogenesis has raised the hypothesis that the new hippocampal neurons may mediate, in part, improved learning associated with exercise. The present review gives an overview of research pertaining to exercise-induced cell genesis, its possible relevance to memory function and the cellular mechanisms that may be involved in this process. PMID- 18286392 TI - Sources of stem cells for regenerative medicine. AB - The shortage of organ donors for regenerative medicine has stimulated research on stem cells as a potential resource for cell-based therapy. Stem cells have been used widely for regenerative medicine applications. The development of innovative methods to generate stem cells from different sources suggests that there may be new alternatives for cell-based therapies. Here, we provide an overview of human embryonic stem cells (hES) and the methods for obtaining these cells and other broadly multipotent or pluripotent cell types. These methods include somatic cell nuclear transfer, single cell embryo biopsy, arrested embryos, altered nuclear transfer, and reprogramming somatic cells. We also discuss the use of amniotic fluid derived stem cells (AFS) for potential patient-specific therapies. PMID- 18286394 TI - The founding of the first psychiatric hospital in the World in Valencia. AB - Several hospitals, countries and cultures claim the privilege of being the first to have provided care to people suffering mental illnesses. Among them, the <> (Hospital of the Innocents) founded in Valencia in 1410 stands out due to its originality and there are historic and cultural reasons to recognize its primacy. Furthermore, the organization and functioning of this institution and the model, spread like wildfire through the entire Iberian Peninsula during the 15th Century and shortly after through American Spanish speaking countries. For centuries, these establishments were considered exemplary and were copied in other European Countries. At the beginning of the 19th Century in Spain a forced sale of the Catholic Church properties or their disamortization among other a large number of hospitals establishments took place. This lead to a terrible collapse of health care for the mentally. From then on it took more than one century to recover a decent standard. The vicissitudes of the creation and progress of the hospital of Valencia and others which followed the example that allows to affirm that it was really the first psychiatric hospital in the World are analyzed in this present work. PMID- 18286393 TI - Targeting cancer stem cells to modulate alternative vascularization mechanisms. AB - Recently, many papers have shown that tumor vascularization can be explained by angiogenesis, recruitment, cooption, vasculogenic mimicry and by mosaic vessels. In particular, vasculogenic mimicry seems to be different from mosaic blood vessels, where tumor cells form a part of the surface of the vessel while the remaining part is covered by endothelium. In this case, tumor cells in apparent contact with the lumen do not show an endothelial phenotype. More recently, vasculogenic mimicry was proposed to occur in patients with multiple myeloma due to bone marrow macrophages. Herein, all these data are, for the first time, discussed critically in comparison to cancer stem cells-which show high trans differentiative capacity-and bone-marrow derived stem cells. In fact, the presence of alternative vasculogenic patterns might be due to the presence of stem cell population (cancer stem cells or bone-marrow stem cells). In this connection, the literature is discussed extensively and possible models are proposed. Pharmacological perspectives will also discuss. PMID- 18286395 TI - Personality disorders from a phenomenological perspective. AB - Different studies have questioned the capacity of the categorical diagnostics to establish a clear distinction between the existence or not of a determined personality disorder. The dimensional perspective would approach more to reality, in the measure that it tries to measure the different intensity degrees in which these disorders are present in the patients. But its application is very laborious and besides, presupposes that those categories whose nuances it pretends to measure really exist. The foresaid leads us to appeal to phenomenological perspective, which seems to be more adequate for the study of complex realities, as it is the case of the personality and its disorders. The essential features of the phenomenological method in the sense of Husserl are described, as well as his contribution to the study of personality disorders. This can be summarized in three fundamental points: the ideal types, introduced in psychiatry by Karl Jaspers, the existential types, by Ludwig Binswanger, and the dialectic typologies and polarities, by Wolfgang Blankenburg and the undersigned. This author defines and develops each one of these concepts, aiming to show their advantages with respect to the categorical and dimensional systems. PMID- 18286396 TI - Spanish version of the Family Interview for Genetic Studies (FIGS). AB - INTRODUCTION: The use of the Family Interview for Genetic Studies (FIGS), has made it possible to advance in the performance of family studies for research in genetic psychiatry. This study aimed to adapt this interview into Spanish and develop a validation process of the FIGS in a sample of Cuban families with a family background of schizophrenia. METHODS: The English version of the FIGS was translated into Spanish and was used in this study. The content was validated through the consensus score of several referees. The questionnaire was administered to at least two informants of 146 families of patients diagnosed of schizophrenia. Internal consistency of the items was analyzed, calculating Pearson's correlation coefficients and Cronbach's alpha of the FIGS and reliability. RESULTS: In general the Spanish version of the FIGS interview was found to be useful. Internal consistency of the FIGS as measured by Cronbach's alpha coefficient has been found to be 0.92 for depression, 0.99 for mania; 0.94 for psychosis, 0.94 for alcohol and drugs and 0.97 for personality disorders, which indicates values having satisfactory reliability. In addition, Pearson's correlation coefficient varied from the 0.41 to 0.99 for the different lists of symptoms mentioned above and all were statistically significant (p<0.0001). CONCLUSIONS: This questionnaire is reliable and valid for gathering diagnostic information about relatives of probands and thus suitable for use in genetic studies of Cuban families with a history of schizophrenia. PMID- 18286397 TI - Effectiveness of mirtazapine in the treatment of depression with associated somatic symptoms. AB - INTRODUCTION: A assess the efficacy of mirtazapine in the treatment of depression with somatic symptoms in a 3-months follow-up study. MATERIAL AND METHODS: DESIGN: multicenter, prospective, observational, open-label, and non controlled study. SAMPLE: seven hundred and eleven patients recruited in outpatient psychiatric consultations by 98 psychiatrists nationwide. INSTRUMENTS: 17-Item Hamilton Depression Rating Scale (HAMD-17) and Standardized Polyvalent Psychiatric Interview (SPPI), somatic symptoms section. Patients were assessed pretreatment and at 15, 30 and 90 days post-treatment. RESULTS: Severity of depression assessed by HAMD-17 significantly decreased (p<0.0001) from 23.27 in the pretreatment assessment to 6.75 at 3 months post-treatment. Severity of somatic symptoms assessed by EPEP significantly decreased (p<0.0001) from 7.68 in the pre-treatment assessment to 2.28 at 3 months post-treatment. Mirtazapine modifies attribution of somatic symptoms in somatizers: in pretreatment assessment, 41.3 % of the sample attributed somatic symptoms to a psychological origin, while at 3 months post-treatment this percentage significantly increased (p<0.05) to 63.94%. Nearly half of the sample (48.52%) took benzodiazepines at the start of the study; but at 3 months post-treatment only 6.71% of the patients needed them. The incidence of adverse effects was 13.36% of the patients. From the total dropouts 4% were due to adverse events. CONCLUSIONS: Mirtazapine is an effective and safe antidepressant for the treatment of depression with somatic symptoms and is able to modify attribution of somatic symptoms in somatizing patients. PMID- 18286398 TI - Intellectual capacity measurement in schizophrenia. AB - INTRODUCTION: The measurement of the intellectual capacity (IC) in schizophrenic patients has been found to be of clinical relevance. A user-friendly tool such as the Cattell's intelligence test might facilitate this measurement in daily clinical practice. METHOD: In this study, we measured the intelligence quotient (IQ) using Cattell's test in 35 schizophrenic patients before and after treatment with risperidone. RESULTS: At baseline, the sample showed an average intelligence of 78.3 points (standard deviation [SD]: 14.3), in the low-medium range. After 1 year on risperidone, the IQ significantly improved (mean: 84.8; SD: 17.0; p = 0.028). This IQ elevation was positively correlated with the improvement in the psychotic symptoms rated with the PANSS. CONCLUSIONS: Cattell's intelligence test could be a valid instrument to measure cognitive performance in schizophrenic patients. Antipsychotic therapy with risperidone could be effective to improve cognitive functioning in these subjects. PMID- 18286399 TI - Psychopharmacological treatment in borderline personality disorder. AB - Borderline personality disorder is a disorder with important social and clinical repercussions, which has been treated mainly by psychotherapy. In recent years, the syndromic analysis of this disorder has allowed us to identify different symptoms capable of being improved with psychopharmacology treatment. Thus, its complex symptomatology could be included in four clinical dimensions: impulsive aggressive, affective instability, cognitive- perceptive and anxiety-inhibition. Antidepressants, mood stabilizers, antipsychotics, anxiolytics, or more recently omega-3 fatty acids have shown efficacy in the treatment of symptomatic dimensions of this disease. We have reviewed scientific articles (reviews, clinical trials or clinical guidelines) published over the last ten years and have proposed therapeutic algorithms for psychopharmacology management in these patients. PMID- 18286400 TI - Sleep evaluation scales and questionaries: a review. AB - INTRODUCTION: Whenever a new scale is created or translated from another language, it must be validated, establishing its reliability for the new population where it will be used. Sleep quality concept is a construct that can be evaluated using self-report scales. Resulting elements vary depending on the individuals surveyed. This type of evaluation is mainly subjective and includes quantitative aspects such as sleep duration, number of awakenings, latency time, and qualitative aspects such as rest sensation, mood and oneiric content (Valencia, 2000). In the present study we made a critical review of the sleep scales designed for child, adolescent and adult populations that have been validated and the difficulties they might present. METHODOLOGY: Between September 2005 and May 2006 a bibliographical search was made within Pubmed, Ovid and the data base of the periodical and book library of the Ramon de la Fuente Muniz National Institute of Psychiatry, using and combining the following key words: sleep, sleep questionnaire, sleep scale, sleep inventory, adolescent, adolescent sleep scale. The most relevant papers to our study were selected. The search was limited to Spanish and English articles, although there was no year or geographical origin limit. Articles that did not include clinimetrical data where excluded. CONCLUSIONS: Based on our bibliographical search and our discussion, we suggested the design and validation of a Spanish scale to evaluate adolescent population which avoids a time interval between awakening and the answering of the instrument in order to decrease recall bias. PMID- 18286401 TI - Effects of smoking ban in a general hospital psychiatric unit. AB - INTRODUCTION: Many psychiatric facilities allow patients to smoke cigarettes even though this habit is especially harmful for mental patients. METHODS: We studied the untoward effects produced by the smoking ban in 40 consecutive inpatients admitted to a psychiatric ward of a general hospital. RESULTS: A total of 52.2 % out of 40 patients were smokers; the mean Fagerstrom score was 5.9. The most frequent diagnostic groups were: schizophrenia and delusional disorders; mood disorders; and personality disorders. There were only two untoward effects related with the smoking ban during the study period. CONCLUSIONS: This study shows that the smoking ban can be successfully implemented in a general hospital psychiatric unit without causing unfavorable effects. PMID- 18286403 TI - Reducing human exposure to aflatoxin through the use of clay: a review. AB - Innovative sorption strategies for the detoxification of aflatoxins have been developed. NovaSil clay (NS) has been shown to prevent aflatoxicosis in a variety of animals when included in their diet. Results have shown that NS clay binds aflatoxins with high affinity and high capacity in the gastrointestinal tract, resulting in a notable reduction in the bioavailability of these toxins without interfering with the utilization of vitamins and other micronutrients. This strategy is being evaluated as a potential remedy for acute aflatoxicosis, and as a sustainable human intervention for aflatoxins via the diet. Phase I and II clinical trials confirmed the apparent safety of NS for further study in humans. A recent study in Ghanaians at high risk for aflatoxicosis has indicated that NS (at a dose level of 0.25%) is effective in decreasing biomarkers of aflatoxin exposure and does not interfere with the levels of serum vitamins A and E, and iron and zinc. In summary, enterosorption strategies/therapies based on NS clay are promising for the management of aflatoxins and as a sustainable public health intervention. The NS clay remedy is novel, inexpensive and easily disseminated. Based on the present research, aflatoxin sequestering clays should be rigorously evaluated in vitro and in vivo, and should meet the following criteria: (1) favourable thermodynamic characteristics of mycotoxin sorption, (2) tolerable levels of priority metals, dioxins/furans and other hazardous contaminants, (3) safety and efficacy in multiple animal species, (4) safety and efficacy in long term studies, and (5) negligible interactions with vitamins, iron and zinc and other micronutrients. PMID- 18286404 TI - Impact of mycotoxins on human health in developing countries. AB - Adverse human health effects from the consumption of mycotoxins have occurred for many centuries. Although mycotoxin contamination of agricultural products still occurs in the developed world, the application of modern agricultural practices and the presence of a legislatively regulated food processing and marketing system have greatly reduced mycotoxin exposure in these populations. At the mycotoxin contamination levels generally found in food products traded in these market economies, adverse human health effects have largely been overcome. However, in the developing world, where climatic and crop storage conditions are frequently conducive to fungal growth and mycotoxin production, much of the population relies on subsistence farming or on unregulated local markets. The extent to which mycotoxins affect human health is difficult to investigate in countries whose health systems lack capacity and in which resources are limited. Aflatoxin B(1), the toxin on which major resources have been expended, has long been linked to liver cancer, yet its other effects, such as immune suppression and growth faltering previously observed in veterinary studies, are only now being investigated and characterized in human populations. The extent to which factors such as immune suppression contribute to the overall burden of infectious disease is difficult to quantify, but is undoubtedly significant. Thus, food safety remains an important opportunity for addressing current health problems in developing countries. PMID- 18286405 TI - Mycotoxin analysis: an update. AB - Mycotoxin contamination of cereals and related products used for feed can cause intoxication, especially in farm animals. Therefore, efficient analytical tools for the qualitative and quantitative analysis of toxic fungal metabolites in feed are required. Current methods usually include an extraction step, a clean-up step to reduce or eliminate unwanted co-extracted matrix components and a separation step with suitably specific detection ability. Quantitative methods of analysis for most mycotoxins use immunoaffinity clean-up with high-performance liquid chromatography (HPLC) separation in combination with UV and/or fluorescence detection. Screening of samples contaminated with mycotoxins is frequently performed by thin layer chromatography (TLC), which yields qualitative or semi quantitative results. Nowadays, enzyme-linked immunosorbent assays (ELISA) are often used for rapid screening. A number of promising methods, such as fluorescence polarization immunoassays, dipsticks, and even newer methods such as biosensors and non-invasive techniques based on infrared spectroscopy, have shown great potential for mycotoxin analysis. Currently, there is a strong trend towards the use of multi-mycotoxin methods for the simultaneous analysis of several of the important Fusarium mycotoxins, which is best achieved by LC-MS/MS (liquid chromatography with tandem mass spectrometry). This review focuses on recent developments in the determination of mycotoxins with a special emphasis on LC-MS/MS and emerging rapid methods. PMID- 18286406 TI - Use of cyclodextrins as modifiers of fluorescence in the detection of mycotoxins. AB - Cyclodextrins, cyclic oligosaccharides composed of amylose subunits, are known to interact with mycotoxins. The interactions may be useful to analytical chemists by altering the properties of the mycotoxin of interest, namely the chromatographic properties, electrophoretic properties, fluorescence, or absorption of these fungal metabolites. Practical applications of these effects have been the incorporation of cyclodextrins into high-performance liquid chromatography and capillary electrophoresis methods for mycotoxin detection. Specific mycotoxins include those with a native fluorescence such as the aflatoxins, ochratoxin A (OTA) and zearalenone (ZEN) as well as those that can be rendered fluorescent through derivatization, such as T-2 toxin. The literature describing the applications of cyclodextrins in mycotoxin analysis is reviewed and an attempt to extend the use of cyclodextrins to the detection of labelled T 2 toxin is presented. Twenty cyclodextrins were evaluated for their ability to enhance the fluorescence emission of T-2 toxin derivatized with pyrene-1-carbonyl cyanide (T2-Pyr). This evaluation revealed that heptakis (2,6-di-O-methyl)-beta cyclodextrin (DIMEB), in particular, enhanced T2-Pyr fluorescence. DIMEB was used as a buffer modifier in a capillary electrophoresis-laser-induced fluorescence (CE-LIF) method for detecting T-2 in maize. Because of the effects that certain cyclodextrins have, especially under aqueous conditions, they may make useful additives for a variety of mycotoxin analytical methods. PMID- 18286407 TI - Mycotoxins in cattle feeds and carry-over to dairy milk: a review. AB - The complex diet of ruminants, consisting of forages, concentrates, and preserved feeds, can be a source of very diverse mycotoxins that contaminate individual feed components. A number of mycotoxins are successfully inactivated by the rumen flora, whereas others pass unchanged or are converted into metabolites that retain biological activity. Hence, the barrier function of the rumen largely determines the susceptibility of dairy cows and other ruminant species towards individual mycotoxins. An impairment of this barrier function due to diseases or the direct antimicrobial effect of certain mycotoxins may increase absorption rates. The rate of absorption determines not only the internal dose and risk for adverse health effects, but also the excretion of mycotoxins and the biologically active metabolites into milk. PMID- 18286408 TI - Mycotoxins in botanicals and dried fruits: a review. AB - Botanicals are used in many countries for medicinal and general health-promoting purposes. Numerous natural occurrences of mycotoxins in botanicals and dried fruits have been reported. Aflatoxins or ochratoxin A (OTA) have been found in botanicals such as ginseng, ginger, liquorice, turmeric, and kava-kava in the USA, Spain, Argentina, India, and some other countries, while fumonisins have been found in medicinal wild plants in South Africa and in herbal tea and medicinal plants in Turkey. Zearalenone was identified in ginseng root. Dried fruits can be contaminated with aflatoxins, OTA, kojic acid, and, occasionally, with patulin or zearalenone. One main area of concern is aflatoxins in dried figs; bright greenish yellow fluorescence under ultraviolet light is associated with aflatoxin contamination. OTA in dried vine fruits (raisins, sultanas, and currants) is another concern. There are also reports of aflatoxins in raisins and OTA in dried figs, apricots, dried plums (prunes), dates, and quince. Maximum permitted levels in the European Union include 4 microg kg(-1) for total aflatoxins in dried fruit intended for direct consumption and 10 microg kg(-1) for OTA in dried vine fruit. This review discusses the occurrence of mycotoxins in botanicals and dried fruits and analytical issues such as sampling, sample preparation, and methods for analysis. Fungal contamination of these products, the influence of sorting, storage, and processing, and prevention are also considered. PMID- 18286409 TI - Managing ochratoxin A risk in the grape-wine food chain. AB - The main source of ochratoxin A (OTA) in the wine food chain is the infection of grapes by "black aspergilli" in the field. OTA-producing black aspergilli include principally Aspergillus carbonarius, followed by A. niger and possibly A. tubingensis. They are opportunistic fungi that develop particularly on damaged berries at ripening, although they may occur and form OTA on grapes from veraison to harvest. Climatic conditions (high humidity and temperature) and geographical location are important factors favouring OTA accumulation in grape berries. The severity of aspergillus rot is influenced by excessive irrigation and rainfall prior to harvest, which causes berry splitting. In addition, berry wounds caused by insect attack provide preferential entries for black aspergilli. High OTA levels occur in grapes severely damaged by the grape moth, Lobesia botrana, particularly in Mediterranean areas. Some grape varieties display greater susceptibility to aspergillus rot due to intrinsic genetic characteristics and bunch conformation (i.e. compact>sparse). Control measures for toxigenic mycoflora in the vineyards must consider these critical control points. Proper fungicidal and insecticidal treatments can reduce OTA contamination. Nevertheless, knowledge about the fate of OTA and its distribution in wine and winery by-products is important to manage OTA risk in contaminated stock. In our wine-making experiments, only 4% of the OTA present in grapes remained in the wine--the majority is retained in pressed grape pomaces. OTA concentration remained unchanged in wine after a 1-year aging as well as in all liquid fractions collected during vinification (i.e. must, free run wine, and wine after first and second decantation). Activated carbon can reduce OTA levels in wine but negatively affects wine quality. PMID- 18286410 TI - Management and prevention of mycotoxins in peanuts. AB - Contamination of peanuts with mycotoxins, particularly aflatoxins, is a worldwide problem that affects both food safety and agricultural economies. Most countries have adopted regulations that limit the quantity of aflatoxins in food and feed to 20 microg kg(-1) or less; however, environmental conditions in most of the world where peanuts are produced and stored often make it difficult or impossible to attain such low concentrations. In addition to aflatoxins, peanuts are often contaminated with cyclopiazonic acid (CPA). Both mycotoxins are produced by Aspergillus flavus, a ubiquitous fungus that can infect and grow in peanuts under both pre- and post-harvest conditions. Management of mycotoxin contamination in peanuts generally involves removal of high-risk components from shelled lots or the removal of individual, highly contaminated nuts. This is accomplished by various processes such as screening, kernel sizing, electronic colour sorting, hand sorting, and blanching followed by electronic colour sorting. Recently, biological control technology has been developed that prevents much of the contamination that might otherwise occur. Biocontrol is based on competitive exclusion whereby a dominant population of a non-toxigenic strain of A. flavus is established in the soil before peanuts are subjected to conditions favouring contamination. The applied strain competes with toxigenic strains for infection sites, resulting in significantly reduced concentrations of aflatoxins in peanuts. Monitoring of the first commercial use of the technology showed that aflatoxins were reduced by an average of 85% in farmers' stock peanuts and by as much as 98% in shelled, edible grade peanuts. PMID- 18286411 TI - Factors influencing fungal and aflatoxin levels in Turkish hazelnuts (Corylus avellana L.) during growth, harvest, drying and storage: a 3-year study. AB - The levels aflatoxins in Turkish hazelnuts have been monitored over a 3-years period (2002-2004). Periodical sampling was made in 72 different orchards at different locations representative of the hazelnut-growing areas and post-harvest applications. Various parameters (aflatoxins, water activity, moulds) were analysed and environmental conditions (temperature and relative humidity) recorded during growing and at different stages of harvest and post-harvest processing, involving three different harvesting methods (collection in nets, from the ground, etc.) and four drying techniques (traditional sun-drying, mechanical drying, etc.). Fungal and aflatoxin analyses (HPLC) showed no significant difference except between samples which had been in contact with the ground and those which had not (at 95% confidence level). Aflatoxins levels from the orchard recorded a maximum of 0.77+/-0.08 ng g(-1) from a total of 1624 samples. Regarding harvesting and post-harvest processes, the only application where aflatoxins were detected was in samples which had been in direct contact with the ground (max. 3.18+/-0.03 ng g(-1)). Aflatoxin formation was low during storage (max. 0.34+/-0.003 ng g(-1)). As a result of mycological studies, a total of 5546 Aspergillus flavus (89%) and A. parasiticus (11%) species were isolated and identified from samples. The results indicated that harvesting hazelnuts into a canvas by shaking the trees, manual harvesting of mature hazelnuts where possible, use of jute instead of nylon sacks and mechanical drying technique would minimize aflatoxin levels in hazelnuts. These recommendations have been implemented and about 4000 people in the hazelnut industry have been trained in these practices. PMID- 18286412 TI - Mycotoxins in small grains and maize: old problems, new challenges. AB - This paper reviews the challenges relating to chronic contamination of small grains and maize with deoxynivalenol and related compounds, fumonisin and the use of ensiled cereals in cool dairy areas. Uncertainties in the tolerable daily intakes for deoxynivalenol and fumonisin are discussed as they have the potential to affect current regulatory limits. In addition, climate change is resulting in more extreme rainfall and drought events which favour formation of deoxynivalenol and fumonisin, respectively. The development and refinement of models for predicting mycotoxin accumulation from weather data will become an essential tool for managing these events. Such models are also important for providing timely food aid to developing countries, which experience increased occurrence of acute toxicities, especially in children. Chronic contamination of silage in some areas with some Penicillium toxins deserves more attention in terms of their economic effects and possible implications for the purity of milk. PMID- 18286413 TI - Design and implementation of an integrated management system for ochratoxin A in the coffee production chain. AB - Coffee is an important export product of Ecuador. Producers are challenged by the implementation of regulatory limits for ochratoxin A. Ecuador has four coffee production areas and the potential for mycotoxin contamination varies due to different environmental conditions and cultural differences in harvesting, storage, processing and commercialization. The major contributors to contamination are the lack of selection during harvesting, delays in drying or rewetting, the lack of proper drying and storage conditions, the mixing of products with different levels of moisture, and the potential for cross contamination. The long commercialization chain involves different intermediaries that use foreign materials to increase the weight of the product without consideration of quality. An integrated mycotoxin management system using the Hazard Analysis Critical Control Point Systems (HACCP) principles was developed to prevent mycotoxin contamination at each stage of production. Critical control points were developed based on the resources available at the different stages of the production chain. Training programmes helped increase awareness about the impact of contamination, but failed to transform knowledge into improved practices. Thus, different demonstrative models specific for each productive region at all production levels were developed to show the application of prevention mechanisms using limited resources and to demonstrate the increased commercial value of coffee produced using good practices throughout the chain so producers have a better disposition to adopt improved practices. Preliminary results show that coffee managed using the models had a better quality, a lower contamination, a higher yield and better commercial value. The use of local resources and low-cost technology was important in demonstrating the practical approach. PMID- 18286414 TI - H.M.'s personal crossword puzzles: understanding memory and language. AB - The amnesic patient H.M. has been solving crossword puzzles nearly all his life. Here, we analysed the linguistic content of 277 of H.M.'s crossword-puzzle solutions. H.M. did not have any unusual difficulties with the orthographic and grammatical components inherent to the puzzles. He exhibited few spelling errors, responded with appropriate parts of speech, and provided answers that were, at times, more convincing to observers than those supplied by the answer keys. These results suggest that H.M.'s lexical word-retrieval skills remain fluid despite his profound anterograde amnesia. Once acquired, the maintenance of written language comprehension and production does not seem to require intact medial temporal lobe structures. PMID- 18286415 TI - Stereotypes influence false memories for imagined events. AB - Two experiments tested the influences of vivid imagery and person schemata on eyewitness accuracy. Participants watched an event sequence including actors performing stereotype-consistent and inconsistent actions. Additionally, participants either read descriptions of actions (Experiment 1) or vividly imagined actions (Experiment 2). After either 30 minutes or 2 days, recognition memory, source memory, and remember/know judgements were made. After 2 days, false alarms to imagined events increased, relative to the 30-minute test; those false alarms were more often misattributed to stereotype-consistent actors, relative to the same actions in the reading condition. In addition, the accompanying remember judgements were higher for false alarms to imagined events, relative to read events, regardless of stereotype consistency. Overall the results suggest that, over time, vivid imagery reinforces schema activation, increasing stereotype-consistent false memories. PMID- 18286416 TI - What if they knocked down the library and nobody noticed? The failure to detect large changes to familiar scenes. AB - The purpose of the current study was to investigate people's ability to detect changes to familiar scenes. College students were asked either to identify what was wrong with a picture of a familiar location on their college campus (e.g., the library had been removed from the scene), or to estimate the difficulty of change detection for a hypothetical cohort performing the same task. Performance in the change-detection condition was extremely poor, even when changes were large. Participants who were familiar with the scenes and those who were unfamiliar with the scenes both overestimated the actual levels of change detection performance. A follow-up analysis indicated that the participants who were unfamiliar with the scenes produced estimations of difficulty that were highly correlated with the mathematical area of the change, whereas participants who were familiar with the scenes produced estimations of difficulty that were highly correlated with the actual difficulty of change detection. The results indicate that people's visual long-term memory for familiar scenes lacks the precision to be able to effectively identify even large-scale changes, although subjectively people believe this should be relatively easy. PMID- 18286417 TI - Optimising self-regulated study: the benefits - and costs - of dropping flashcards. AB - Self-regulation of study activities is a constant in the lives of students - who must decide what to study, when to study, how long to study, and by what method to study. We investigated self-regulation in the context of a common study method: flashcards. In four experiments we examined the basis and effectiveness of a metacognitive strategy adopted almost universally by students: setting aside (dropping) items they think they know. Dropping has a compelling logic - it creates additional opportunities to study undropped items - but it rests on two shaky foundations: students' metacognitive monitoring and the value they assign to further study. In fact, being allowed to drop flashcards had small but consistently negative effects on learning. The results suggest that the effectiveness of self-regulated study depends on both the accuracy of metacognitive monitoring and the learner's understanding, or lack thereof, of how people learn. PMID- 18286418 TI - Changing the criterion for memory conformity in free recall and recognition. AB - People's responses during memory studies are affected by what other people say. This memory conformity effect has been shown in both free recall and recognition. Here we examine whether accurate, inaccurate, and suggested answers are affected similarly when the response criterion is varied. In the first study, participants saw four pictures of detailed scenes and then discussed the content of these scenes with another participant who saw the same scenes, but with a couple of details changed. Participants were either told to recall everything they could and not to worry about making mistakes (lenient), or only to recall items if they were sure that they were accurate (strict). The strict instructions reduced the amount of inaccurate information reported that the other person suggested, but also reduced the number of accurate details recalled. In the second study, participants were shown a large set of faces and then their memory recognition was tested with a confederate on these and fillers. Here also, the criterion manipulation shifted both accurate and inaccurate responses, and those suggested by the confederate. The results are largely consistent with a shift in response criterion affecting accurate, inaccurate, and suggested information. In addition we varied the level of secrecy in the participants' responses. The effects of secrecy were complex and depended on the level of response criterion. Implications for interviewing eyewitnesses and line-ups are discussed. PMID- 18286419 TI - Compelling untruths: the effect of retention interval on content borrowing and vivid false memories. AB - False memories are sometimes vivid and detailed. We proposed that the details that come to be associated with false memories may be produced by a process called content borrowing. In content borrowing, details from presented items are erroneously borrowed to corroborate the occurrence of the false memory item. In the present research we used think-out-loud protocols at both study and test to provide evidence for content borrowing in the DRM paradigm. We also demonstrated that content borrowing was more common after a 2-week delay, and was more common for critical lures than for missing exemplars. PMID- 18286420 TI - Does the presence of priming hinder subsequent recognition or recall performance? AB - Declarative and non-declarative memories are thought be supported by two distinct memory systems that are often posited not to interact. However, Wagner, Maril, and Schacter (2000a) reported that at the time priming was assessed, greater behavioural and neural priming was associated with lower levels of subsequent recognition memory, demonstrating an interaction between declarative and non declarative memory. We examined this finding using a similar paradigm, in which participants made the same or different semantic word judgements following a short or long lag and subsequent memory test. We found a similar overall pattern of results, with greater behavioural priming associated with a decrease in recognition and recall performance. However, neither various within-participant nor various between-participant analyses revealed significant correlations between priming and subsequent memory performance. These data suggest that both lag and task have effects on priming and declarative memory performance, but that they are largely independent and occur in parallel. PMID- 18286422 TI - Reliability and validity assessment of the hand activity level threshold limit value and strain index using expert ratings of mono-task jobs. AB - This study evaluated two subjective assessment methods for physical work-related musculoskeletal disorder (WMSD) risk factors. A total of 567 participants from 12 companies in the manufacturing and health care industries were evaluated using the hand activity level (HAL) threshold limit value (TLV) and the Strain Index. Inter-rater reliability comparisons were performed on 125 selected cyclic tasks, with one novice and three experienced raters. Predictive validity was assessed by evaluation of relationships between measured exposure parameters and diagnosed WMSDs of the hand/wrist and elbow. HAL hand repetition ratings had a Spearman r value of 0.65 and a kappa value of 0.44 between raters. Subjective force (0-10 scale) estimates had a Spearman r = 0.28 and were not significantly different between raters (p > .05). The rating comparison for the four subjective components of the Strain Index had Spearman r correlations of 0.37-0.62 and kappa values of 0.25-0.44. The Strain Index and HAL TLV agreed on exposure categorization 56% of the time. Logistic regression showed, after adjustment for age, gender, and body mass index, that higher peak hand force estimates (odds ratio [OR] 1.14, confidence interval (CI) 1.02-1.27), most common force estimates (OR 1.14, CI 1.02-1.28), hand/wrist posture rating (OR 1.71, CI 1.15-2.56), and Strain Index scores >/= 7 (OR 1.82, CI 1.04-3.18) were associated with distal upper extremity disorders in the dominant hand. HAL repetition ratings >/= 4 (OR 2.81, CI 1.40-5.62) and hand/wrist posture ratings (OR 1.59, CI 1.01-2.49) were associated with disorders in the nondominant hand. These findings show moderate to good inter-rater agreement and significant relationships to health outcomes for the identified measures. PMID- 18286423 TI - A moving robotic hand system for whole-glove permeation and penetration: captan and nitrile gloves. AB - The aim of this study was to develop a robotic hand to test the influence of hand movement on the permeation/penetration of captan through disposable nitrile rubber gloves. An available robotic hand was modified to within one standard deviation of the anthropometric 50th percentile male hand. Permeation tests used a nylon inspection glove interposed between medium-size outer and inner nitrile gloves, the latter protected the hand. Permeation of an aqueous emulsion (217 mg/mL) of captan was conducted at 35 degrees C +/- 0.7 degrees C. A new surface wipe technique facilitated collection of captan from the inner surface of the exposed nitrile gloves, a technique favored above rinse methods that extracted captan from within the glove. With hand movement, the permeated mass of captan collected after 8 hr ranged from 1.6 to 970 microg (Brand A) and 8.6 +/- 1.2 microg (Brand B). Without hand movement, the corresponding masses ranged from 1.4 to 8.4 microg (Brand A) and 11 +/- 3 mg (Brand B). These results were not significantly different at p < or = 0.05 using parametric and nonparametric statistical tests but indicated that hand movement could influence the precision of permeation (F-test p < or = 0.05). One glove exhibited failure after 2 hr with movement, in comparison with 0.5 to 9.9 microg captan with no movement. Hand movement did not appear to significantly affect the permeation of captan through nitrile gloves. However, hand movement did influence physical and/or chemical degradation, resulting in glove failures. The robotic hand simulated normal hand motions, was reliable, and could be used to assess the influence of hand movement on the permeation of nonvolatile components through gloves. Future research should continue to investigate the influence of hand movement and additional work factors on the permeation, penetration, and physical integrity of protective gloves. PMID- 18286424 TI - An efficient analytical method for particle counting in evaluating airborne infectious isolation containment using fluorescent microspheres. AB - The containment performance of patient isolation enclosures, particularly expedient surge capacity enclosures, must be verified to protect health care providers and staff, other patients, and hospital visitors. Tracer gas methods are often used, but requirements for special equipment and training limit the technique's utility. A technologically simple yet accurate and precise particle based technique is needed to measure the low count concentrations of escaping airborne particles that might be present outside an isolation enclosure. Reported here is the performance of such a technique employing micrometer-sized fluorescent polystyrene latex microspheres as a surrogate for pathogenic bioaerosols. Particles are released into the isolation enclosure, air is sampled inside and outside the room to capture airborne particles on 25 mm diameter filters, and the number of particles deposited on a filter is quantified using an optimized random field counting approach. The technique accurately estimates the number of surrogate bioaerosol particles on the filter, allowing calculation of the airborne particle concentrations inside and outside the enclosure, and the containment efficiency. This technique can be employed using generally available equipment and inexpensive supplies and also can minimize the number of particle counts that must be performed. The method is shown to be specific, sensitive, and accurate. PMID- 18286425 TI - Octyl-methoxycinnamate (OMC), an ultraviolet (UV) filter, alters LHRH and amino acid neurotransmitters release from hypothalamus of immature rats. AB - OMC (octyl-methoxycinnamate), is an endocrine disruptor with estrogenic activity, which is used in sunscreen creams as a UV filter. We studied its " IN VITRO" effects on the hypothalamic release of LHRH as well as on the amino acid neurotransmitter system in immature rats of 15 (prepubertal) and 30 (peripubertal) days of age. OMC decreased the LH-RH release significantly in male and female rats of both age. In male rats OMC increased the release of GABA while in the female ones It diminished the excitatory amino acid aspartate (ASP) and Glutamate (GLU) without modifications in the hypothalamic GABA release. These results suggest that during sexual maturation the inhibitory effect of OMC on LH RH release appears to be related to its action on the inhibitory and excitatory amino acid neurotransmitters in male and female rats. PMID- 18286426 TI - Bone marrow mesenchymal stem cells induce angiogenesis and attenuate the remodeling of diabetic cardiomyopathy. AB - Independent of the severity of coronary artery disease, diabetic patients have an increased risk of developing heart failure. Diabetic cardiomyopathy (DCM) is characterized by microvascular pathologies and interstitial fibrosis. Mesenchymal stem cells (MSCs) are pluripotent and are able to differentiate into cardiomyocytes and vascular endothelial cells. Studies have demonstrated MSCs transplantation can prevent apoptosis of ischemic heart via upregulation of Akt and eNOS and inhibit myocardial fibrosis of dilated cardiomyopathy by decreasing the expression of matrix metalloproteinase (MMP) in rat models. In order to find out whether transplantation of MSCs is a promising treatment in DCM, we used streptozotocin (STZ) -induced diabetic rats as the model. Exogenous MSCs were injected into the femoral vein 8 weeks after STZ injection. Using independent experimental approaches, we showed that MSCs presented in the myocardium 4 weeks after transplantation and some of them were positive for the cardiac markers Troponin T and myosin heavy chain. MSCs transplantation significantly increased myocardial arteriolar density and decreased the collagen volume in diabetic myocardium resulting in improved cardiac function. Furthermore, MSCs transplantation increased MMP-2 activity and decreased transcriptional level of MMP-9. These results show that MSCs transplantation improved cardiac function in the rat DCM model, possibly through angiogenesis and attenuation of cardiac remodeling. PMID- 18286428 TI - Aesthetic reconstruction of head and neck defects. Preface. PMID- 18286427 TI - Acute hyperglycemia rapidly suppresses endothelium-dependent arterial dilation in first-degree relatives of type 2 diabetic patients. AB - OBJECTIVE: Previous studies showed that endothelium-dependent arterial dilation is impaired in first-degree relatives of type 2 diabetes in the fasting state. In the present study, we examined whether endothelial dysfunction occurs when acute hyperglycemia is induced by oral glucose loading in this cohort. PATIENTS AND METHODS: This study included 32 normal glucose tolerant subjects. Of them, 17 with a family history (FH) of type 2 diabetic parents (FH+) and 15 with no first degree relative with diabetes or coronary artery disease (FH-). The examination of vascular function was performed in fasting state and repeated 1 and 2 hours after a 75-g oral glucose loading by high resolution ultrasound. RESULTS: Endothelium- dependent arterial dilation in FH+ group were significantly lower than those in FH- before and after oral glucose loading (5.12+/-0.61% vs 6.03+/ 0.56%, fasting; 4.0+/-0.65% vs 5.70+/-0.42%, 1 h; 4.43+/-0.61% vs 5.82+/-0.67% 2 h, p<0.05 each). In FH+ group, endothelium-dependent arterial dilation decreased significantly at 60 min (4.0+/-0.65% vs 5.12+/-0.61%, p<0.01) and increased markedly from 60 min at 120 min (4.43+/-0.61% vs 4.0+/-0.65%, p<0.05), which was still significantly lower than baseline (4.43+/-0.61% vs 5.12+/-0.61%, p<0.01) . In FH- group, however, the arterial dilation did not differ significantly among the three time points (p 0.05). In multiple regression analysis, endothelium dependent arterial dilation was significantly correlated to FH+(r=-0.302, p<0.01). In addition, endothelium-dependent arterial dilation showed a correlation with plasma glucose (r=-0.460, p<0.01) and TBARS (r=-0.382, p<0.01) during OGTT in FH+ subjects. CONCLUSION: Significant endothelial dysfunction is present in the fasting state, hyperglycemia in response to oral glucose loading rapidly suppresses endothelium-dependent arterial dilation in FH+ subjects, probably through increased production of oxygen-derived free radicals. PMID- 18286429 TI - The path of nasal reconstruction: from ancient India to the present. AB - Nasal reconstruction, first described over 2600 years ago in India, has undergone several modifications in its development. However, there is a misconception that this surgery began with the forehead flap repair, and the history of the subsequent two and a half millennia is poorly represented in most surgical texts. This article presents a review of the historical significance of nasal injuries in antiquity and the various technical developments along the path to modern nasal reconstruction, from ancient India through medieval Europe to modern England and America. Although written texts from 6th-century BC India through 16th-century AD Europe described pedicle flap repair of nasal defects, it was not until the late 18th century that the first written description of the forehead flap appeared. Forehead flap repair developed on an alternate pathway, being transmitted via an oral tradition, typically within families of craftsmen, at least as early as the 14th century. These ancient authors recognized the need for accurate flap design and sizing, donor site repair, precise tissue apposition, protection of the flap pedicle, hemostasis, deepithelialization of the wound, and replacement of intranasal lining. By appreciating ancient surgical techniques and examining the particular issues these procedures attempted to address, the surgeon will gain an additional and more detailed perspective of nasal reconstruction that will aid in providing excellent care to his patients. PMID- 18286430 TI - Aesthetic considerations in scalp reconstruction. AB - This article reviews common methods of reconstructive surgery in patients with wounds that involve the scalp, including primary wound repair, healing by secondary intention, and the use of skin grafts, local tissue flaps, regional myocutaneous flaps, and microvascular free flaps. Special attention is paid toward consideration of aspects of the reconstruction that affect the aesthetic outcome, including preservation of the hairline and hair follicle orientation, scar camouflage, avoidance of alopecia, and secondary restoration of alopecia. PMID- 18286431 TI - Computer-designed implants for fronto-orbital defect reconstruction. AB - Patients with complex defects of the frontal and orbital regions present a myriad of problems for the reconstructive surgeon. In this review, options that allow state-of-the-art reconstruction using computer-assisted implants will be illustrated. The advantages of such implants created with computer assistance will be reviewed as well as indications for their use. PMID- 18286432 TI - Aesthetic considerations in mandibular reconstruction. AB - As free tissue transfer methods have improved, vascular bone grafting has become state of the art for reconstruction of mandibular defects. Prior studies have focused on flap survival and functional outcomes. The reconstructive surgeon should also strive to attain lofty aesthetic goals for this group of patients. The best results are achieved when patient factors, flap selection, treatment planning, and surgical techniques are all considered and properly selected. PMID- 18286433 TI - Incisionless tarsal-strip, canthoplasty, and oral commissureplasty procedures for correction of facial nerve paralysis. AB - Facial Nerve (FN) paralysis and its morbidities can result from a wide variety of etiologies. Several novel surgical procedure methods for remediation of FN paralysis problems, using Incisionless technique are presented. The procedures are minimally-invasive and well-tolerated by patients. The Incisionless procedures achieve basic resuspension of the major orbital and oral morbidities associated with FN paralysis. PMID- 18286434 TI - Management of cutaneous juvenile hemangiomas. AB - Past descriptions of vascular lesions often confused vascular malformations and hemangiomas using interchangeable definitions which led to inappropriate treatment and inconsistency in the medical literature. The work of Mulliken and Glowacki systematically delineated the difference between hemangiomas and the multiple varieties of vascular malformations. Hemangiomas are in fact as the suffix "oma" suggests true tumors exhibiting cellular proliferation on histology. Additionally, treatment has been clouded by numerous reports in the literature of near universal complete spontaneous resolution and conversely arguments that every lesion demands treatment. More recently, thoughtful studies of the natural course of hemangiomas have been completed, algorithms for intervention versus observation proposed, and effective safe treatment plans devised. The following article will be presented in two sections. The first section will review the diagnosis and natural history of hemangiomas as appreciated in modern literature and compare this to a review of historical articles. This information will then be used to describe a rational and thoughtful algorithm for observation or intervention and recommend appropriate treatment options. Special emphasis will be given to surgical technique and several cases of late involuting hemangiomas of the face will be presented. PMID- 18286435 TI - Aesthetic management of external skin paddles following microvascular reconstruction of the head and neck. AB - Microvascular free tissue transfer has become a popular reconstructive option for a wide range of complex head and neck defects. Although highly effective in achieving wound closure, this technique frequently introduces skin paddles with inherent differences in thickness, color, and texture. In resurfacing of cutaneous facial defects, such disparity often leads to inferior aesthetic results. For patients, poor appearance following head and neck reconstruction can become a significant source of distress and add additional burden to their recovery. This article describes several options available in the postoperative setting to improve the appearance of transferred skin paddles. Regional tissue rearrangement with the submental and posterior scalping flaps is discussed in detail. PMID- 18286436 TI - Aesthetic and functional management of eyelid and orbital reconstruction. AB - Major head and neck resections may result in ocular defects that are functionally and/or aesthetically incapacitating. Restoration of the eyelid and orbit must address lateral canthal laxity, midface ptosis, eyelid retraction and ptosis, globe malposition, and dysfunctional lacrimal drainage. Here we discuss lateral canthal reconstruction, midface-lifting, eyelid spacer grafts, gold weight placement, surgical approaches to the orbit, free flap options for orbital reconstruction, and endoscopic lacrimal surgery. Successful outcomes in eyelid and orbital reconstruction depend upon proper knowledge, planning, and multidisciplinary management. PMID- 18286437 TI - Endoscopic treatment of facial fractures. AB - The application of endoscopic surgical techniques to the treatment of craniomaxillofacial fractures not only has decreased the morbidity associated with the surgical approaches but has significantly altered the treatment philosophy for many types of fracture. Frontal sinus fractures, orbital floor fractures, zygomatic arch fractures, and subcondylar mandible fractures are the most notable examples where endoscopic techniques have found acceptance. PMID- 18286438 TI - Advanced lip reconstruction: functional and aesthetic considerations. AB - The lips have both aesthetic and functional features that serve to provide recognizable individualized beauty, emotional cues, sensual interactions, speech abilities, and oral continence for nutrition. Lip distortion or loss occurring from trauma or neoplasms can have devastating cosmetic and form deficits with resultant psychological, physical, and nutritional detriments. Appropriate reconstruction of the lips requires a balance between form, function, and aesthetics. Conservation of tissue when feasible should be the overall goal followed by exhausting and maximizing on all adjacent local tissue advancements, rotations, and transposition options. Tissue preservation as the first line of lip reconstruction will achieve the highest success in maintaining sphincteric function along with balancing appearance. This goal should be maintained when dealing with simple lacerations with minimal tissue loss to extensive near or total lip defects. We hope to provide an algorithm and review of aesthetic considerations in lip reconstructive techniques for a wide range of lip defects. An anatomic review and historical background followed by aesthetic issues and pearls related to defect size-dependent lip reconstruction techniques will be presented. The emphasis will be on cosmetic issues that arise with lip reconstruction and how to incorporate a detailed preoperative assessment; minimize donor site morbidity; match tissue size, texture, and color; and maintain stomal competence to balance form, function, and beauty. PMID- 18286439 TI - Aesthetic repair of small to medium-sized nasal defects. AB - Reconstruction of small and medium-sized defects of the nose poses a challenge to the facial plastic surgeon. Flaps for small to medium-sized defects most often are closed in single-staged procedures as opposed to larger-sized defects. A variety of techniques can be used including secondary intention, primary closure, full-thickness skin grafts, composite grafts, rhomboid flaps, bilobe flaps, dorsal nasal flaps, island flaps, and inferiorly based meliolabial flaps. PMID- 18286440 TI - Aesthetic microtia reconstruction with Medpor. AB - The complex architecture of the auricle makes it one of the most challenging structures for the reconstructive surgeon to re-create. Overlying the ear's unique cartilage framework are layers of varied soft tissues forming a three dimensional organ, which is distinctively positioned on the head. Arguably, the most challenging auricle to reconstruct is third-degree microtia due to a near total absence of native tissue and a need for lifelong durability of the reconstruction. Many methods of reconstruction have been studied; autogenous costal cartilage reconstruction has been one of the more traditional methods, with favorable long-term results reported by several surgeons. However, this technique requires tremendous artistic and technical skill on the part of the surgeon-sculptor to construct a realistic-appearing ear. High-density porous polyethylene (Medpor) is a stable, alloplastic implant that can integrate with host tissues, is resistant to infection, and has been successfully applied to reconstruction of the head and neck. For auricular reconstruction, Medpor- enveloped in a temporoparietal fascial flap with full-thickness skin graft coverage--is a durable and aesthetically gratifying alternative in microtic patients. This alternative surgical technique reduces surgical time and morbidity, standardizes results among surgeons, and facilitates an aesthetic, natural-appearing reconstruction of the auricle. PMID- 18286441 TI - Advances in approaches to the cranial base: minimizing morbidity. AB - The most innovative and meaningful recent advances regarding surgery of the cranial base involve the ability to perform a complete resection followed by a water- and airtight reconstruction while minimizing facial incisions and morbidity. Perhaps the first step in this direction took place when the subcranial/subfrontal approach was introduced for anterior skull base surgery. Originally developed by Raveh in 1978 for the management of severe skull base injuries, these approaches were later adapted for the treatment of congenital anomalies prior to their utilization for resection of anterior skull base tumors. The endoscopic approaches are quite practical with promising long-term efficacy for the treatment of most benign, infectious, and inflammatory disorders. The minimal recovery time, functional outcomes, and obvious aesthetic advantages are only tempered by the lack of long-term data regarding the efficacy of these approaches in the treatment of malignancies. PMID- 18286442 TI - Advances in cranioplasty: a simplified algorithm to guide cranial reconstruction of acquired defects. AB - The search for a universal reconstructive material and the application of a routine approach leaves the surgeon limited in cranial reconstruction. Facility with a variety of implant materials and approaches helps optimize surgical management. The variability of surgical defects and challenges in cranial reconstruction highlights the importance of a strategic approach to evaluate both the defect and the clinical circumstance. Among the numerous alloplastic materials available, the utility of titanium mesh, hydroxyapatite cement, and prefabricated custom acrylic implants have remained at the forefront. The onus of responsibility remains on the surgeon to evaluate each clinical scenario and select the optimal course for management. PMID- 18286443 TI - Pearls for aesthetic reconstruction of cleft lip and nose defects. AB - Mastery of the anatomy and embryology of the normal and cleft upper lip, primary palate, and secondary palate is an essential foundation for grasping techniques in aesthetic reconstruction of cleft lip defects. The surgical goals in repairing cleft lip deformities are to address the deficiencies of the cleft lip defect, restore static and dynamic anatomy, reshape the cleft nasal deformity, and leave a natural-appearing scar that mimics the contours of the philtral components. An additional goal is to improve skeletal alignment and retention of teeth in the vicinity of the alveolar cleft. There are advantages and disadvantages inherit in all repair techniques. However, there are principles that can be universally applied and that will improve the success of most approaches. Certain steps deserve special attention to detail, which provide for enhanced results in lip repair. In addition, the pearls for aesthetic reconstruction of cleft lip and nose defects outlined within this article--attention to scars and surface detail, utilizing the orthopedic forces of the orbicularis pull to achieve improved alveolar alignment, the importance of mucosal detail, and attention to the cleft nasal defect--will help to improve results and reduce secondary defects. PMID- 18286444 TI - Mental health, health, and substance abuse service needs for the Native American Rehabilitation Association Northwest (NARA NW) in the Portland, Oregon metropolitan area. AB - Consistent with results of previous needs assessments for urban American Indian and Alaska Native populations, a needs assessment in the Portland,Oregon metropolitan area for the Native American Rehabilitation Association Northwest revealed high levels of co-occurring conditions for American Indian and Alaska Native clients, often combining chronic health problems, substance abuse histories, and mental health diagnoses. Focus group results suggest the need for crisis care as well as specific needs of children and families, veterans, elders, and adults. PMID- 18286445 TI - Predictors of relapse for american Indian women after substance abuse treatment. AB - The objective of this study was to describe the predictors of substance use relapse of American Indian (AI) women up to one year following substance abuse treatment. Relapse is defi ned as any use of alcohol or drugs in the past 30 days at the follow-up points. Data were collected from AI women in a 45-day residential substance abuse treatment program. Predictors include distal (in time) proximal (recent), and intrapersonal factors. Results indicated that intrapersonal factors showed the strongest relationship with relapse, followed by proximal and distal factors. Negative messages about using alcohol or drugs from the client's father while growing up may have had an impact on whether the client used alcohol at 6 months. Confl icts with other people and being in the company of alcohol or drug users were highly predictive of relapse. While craving was highly predictive of substance use at follow up, self-efficacy was highly predictive of no substance use. Knowledge about predictors of relapse among this population should be used as a guide toward individual treatment planning. PMID- 18286446 TI - Investigation of factors contributing to diabetes risk in american indian/alaska native youth. AB - This study investigated the relationship between family history, sedentary behaviors, and childhood risk for type 2 diabetes. Participants were 480 students attending schools on or near an American Indian reservation. Data were collected through survey and BMI measurement. Children who frequently watched television or played video games did not significantly differ in BMI compared to peers. However, children with a parental history of diabetes had significantly higher BMIs than children without. PMID- 18286447 TI - The gambling behavior of American Indian and non-Indian participants: effects of the actions and ethnicity of a confederate. AB - The present experiment investigated whether the gambling of American Indian (AI) and non-AI participants would be sensitive to the actions and/or ethnicity of another gambler (i.e., a confederate) when playing a slot-machine simulation. Eight male AIs and eight male non-AIs participated in five gambling sessions. In one, the participant gambled alone. In the other four, the participant played in the presence of a confederate of the same or different ethnicity who gambled the entire session or quit after playing five times. The gambling of the AI and non AI participants did not differ, nor was either group sensitive to whether the confederate was AI or non-AI. Gambling behavior was altered by the confederate's actions, with participants gambling less when the confederate left the session than when alone or when the confederate stayed and gambled. These results suggest that the differences in gambling problems between AIs and non-AIs reported in the overall literature may not be a function of ethnicity per se. They also suggest that the actions of other gamblers may inhibit gambling, which may have treatment implications. PMID- 18286448 TI - Cardiorenal and renocardiac syndromes: clinical disorders in search of a systematic definition. PMID- 18286449 TI - Potential adverse effects of replacing high volume hemofiltration exchanges on electrolyte balance and acid-base status using the current commercially available replacement solutions in patients with acute renal failure. AB - The original substitution and/or dialysates used for continuous hemofiltration and/or dialysis were either based on peritoneal dialysis fluids, or peritoneal dialysates themselves. Although these fluids have been refined over the years, by changing the anionic base from acetate or racemic d+ l-lactate, to bicarbonate or l-lactate, the fluids remain designed for low volume hourly exchanges. Even at relatively low volume exchanges, the composition of these fluids can affect systemic acidbase balance. Those fluids which contain a higher concentration of lactate with a corresponding lower chloride, predispose to a hypochloremic alkalosis, whereas those with a lower concentration of lactate with a corresponding higher chloride potentially lead to a hyperchloremic acidosis. In addition, cation balance, differs when using the same fluid as a replacement solution, compared to dialysate. In particular, sodium balance is greater for post-dilutional fluid replacement, due to the lower sieving coefficient. More recently, citrate has been introduced as an extracorporeal anticoagulant, and a variety of dialysates/replacement fluids have been developed, due to the difference in citrate clearance between dialysis and convection based extracorporeal circuits. Over the last decade, there has been a general increase in the standard hourly dialysate and exchange volumes used in the management of patients with acute kidney injury, from 1 to 2.5-3 l/h. Indeed, even larger volume exchanges, some 6-8 l/h have been advocated to improve patient outcomes. However, before embarking on such continuous high volume exchanges, a new generation of fluids may need to be developed to prevent acid-base and electrolyte imbalances. PMID- 18286450 TI - Acid-base disturbances in the intensive care unit: current issues and the use of continuous renal replacement therapy as a customized treatment tool. AB - Continuous renal replacement therapies (CRRT) are often used to manage complex acid-base problems in critically ill patients. These techniques allow a constant manipulation of the plasma composition. Several technical factors from CRRT influence the acid-base status; namely, the effluent rate, the operational characteristics of the technique, the content of the solutions and the metabolic rate of the buffer. This article reviews the common acid base disorders occurring in the intensive care unit, using both the anion gap and the strong ion gap approaches, and describes the influence of CRRT on acid-base physiology. The use of CRRT as a customized therapy for acid-base disorders is discussed, allowing an integration of both physiological and technical concepts. PMID- 18286451 TI - Hepatitis C infection in patients with chronic kidney disease. AB - The management of hepatitis C virus (HCV)-infected patients with chronic kidney disease (CKD) is complex and represents a particular concern since numerous issues, such as antiviral therapy in dialysis patients and post renal transplant, and prevention of HCV spread within dialysis units, remain unresolved. An enormous body of literature has been published on HCV in the CKD population; however, clinical evidence on important issues is mostly based on uncontrolled clinical trials or retrospective surveys. The aim of this paper is to provide a systematic review of the literature. Responses to the critical issues have been developed by a consensus of experts, endorsed by the Italian Association for the Study of the Liver (AISF) and some clinical recommendations have been added. PMID- 18286452 TI - Impaired brachial artery endothelial flow-mediated dilation and orthostatic stress in hemodialysis patients. AB - PURPOSE: Chronic kidney disease (CKD) is associated with an impaired endothelial function, which may contribute to cardiovascular events. Whether impairment in endothelial function is involved in the circulatory response to orthostatic stress is unknown. We assessed endothelial function via brachial artery flow mediated dilation (BAFMD), an index of endothelial-dependent vasodilation. METHODS: We measured changes in brachial artery diameter (BAD) and blood flow by Doppler ultrasound in 35 CKD patients on hemodialysis, 37 young healthy controls (HC) and 50 non-uremic matched controls (MC), in the supine position and after 60 degrees head-up tilting (HUT). RESULTS: In the supine position, endothelial flow mediated BAD was significantly increased in HC (p<0.001) and MC (p<0.01) while no significant changes were detected in CKD. Mean percent blood flow changes were HC+323.5%, MC+195.1% and CKD+158.8% (HC vs. CKD p<0.001; HC vs. MC p<0.001; MC vs. CKD p=0.04). Similarly, during HUT mean BAD and blood flow increases were significantly impaired in CKD patients. CONCLUSION: In CKD patients, an impaired response in the physiologic vascular reactivity, suggesting endothelial dysfunction, was found in the supine position and after orthostasis by BAFMD. Our results are in favor of a possible adjunctive role of uremia in the abnormal brachial artery response. PMID- 18286453 TI - Citrate anticoagulation protocol for slow extended hemodialysis with the Genius dialysis system in acute renal failure. AB - BACKGROUND: The Genius dialysis system is increasingly used as an intermittent hemodialysis device in the setting of acute renal failure. Slow extended hemodialysis is preferred in the case of critical ill patients. In this study we established a safe and feasible citrate anticoagulation protocol for slow extended hemodialysis (SLED) with the Genius system. METHODS: We compared six anticoagulation protocols using SLED in 34 critically ill patients with acute renal failure. One group (A) received only citrate anticoagulation. Four groups (B - D) were treated with citrate and different additional systemic anticoagulation. Patients in the last group (F) were anticoagulated with heparin and were free of citrate anticoagulation. The total number of treatments was 103. A 4% sodium citrate solution was infused into the arterial line of the dialysis device for citrate anticoagulation. The dialysis solution contained one mmol/L of calcium. No additional calcium supplementation was done. We monitored electrolyte, acid-base and cardiovascular status prospectively. RESULTS: Hemodialysis was well tolerated hemodynamically. Electrolytes remained stable throughout hemodialysis in all groups. The decrease in ionized and total calcium was within the expected, clinically acceptable range. Bicarbonate and pH levels increased during dialysis, especially if citrate was used. CONCLUSIONS: Slow extended Genius hemodialysis with citrate is well tolerated and offers a safe and effective alternative to systemic anticoagulation. PMID- 18286454 TI - Peritoneal dialysis-related peritonitis in southern Thailand. AB - OBJECTIVE: To evaluate peritonitis in children receiving peritoneal dialysis (PD) in southern Thailand. PATIENTS AND METHODS: We reviewed the records of patients who received PD at the Department of Pediatrics, Prince of Songkla University from January 1994 to December 2003. RESULTS: Forty-six patients had PD performed for 228.6 patient-months. Their mean age was 9.5+/-4.8 years (range 3.0 months 16.7 years). Twenty-eight patients had complications with 61 episodes of peritonitis. The age, sex and causes of renal failure did not display any differences between patients who had or did not have peritonitis (p=0.2, 0.6 and 0.6, respectively). The first peritonitis occurrence was on average at 2.7+/-4.0 patient-months (median 0.4, range 0-12.5) following catheter insertion, with an average incidence rate of one infection per 3.5+/-4.8 patientmonths (median 1.0, range 0-23.2). The causative agents were both gram-positive and gram negative bacteria, and fungi at 24%, 44%, and 8% respectively (24% of the cultures did not grow). There was no significant difference in causative agents between home- and hospital-acquired peritonitis (p=1.0). CONCLUSION: PD-related peritonitis in this study occurred earlier and more often than in other studies, probably because all of our PDs were performed immediately after catheter insertion. PMID- 18286455 TI - Evaluation of a hybrid artificial liver module with liver lobule-like structure in rats with liver failure. AB - We studied the recovery of rats with fulminant hepatic failure (FHF) by treating them with our original hybrid artificial liver support system (HALSS). We developed an original artificial liver module having a liver lobule-like structure (LLS). This module consists of many hollow fibers regularly arranged in close proximity and hepatocyte aggregates (organoids) induced into the extra capillary space of the module by centrifugal force. The LLS module can express some liver specific functions at high levels and maintain them for several months in vitro. In this study, we evaluated the efficacy of our LLS-HALSS by using rats with FHF induced by a method that combined partial hepatectomy with hepatic ischemia. In the animal experiments, blood ammonia levels rapidly increased in the control group (sham-HALSS group). These rats died during or immediately after application of the sham-HALLS. On the other hand, in the LLS module application group (LLS-control group), the increase in blood ammonia was completely suppressed and all rats recovered. Blood constituents at 4 weeks after application were at normal levels, and the weight of the liver was the same as that of a normal rat. These results indicate that HALSS may be useful for treating liver failure patients until liver transplantation can be performed or until regeneration of the native liver occurs. PMID- 18286456 TI - Uterine myometrium as a cell patch as an alternative graft for transplantation to infarcted cardiac myocardium: a preliminary study. AB - PURPOSE: Currently, only a small fraction of patients are able to receive reperfusion therapy for myocardial infarctions. We hypothesize that myometrial cell patch transplantation could be an alternative approach for the treatment of myocardial infarction. DESIGN: We performed a preliminary study to determine the feasibility of this novel therapeutic approach in a rabbit model. PROCEDURES: Six adult female New Zealand rabbits were used. Myocardial infarction was induced by left anterior descending artery ligation. A segment of uterus was removed via a laparotomy incision, and this uterine segment was transplanted as an autologous graft over the infarcted myocardium, which was then reinforced by greater omentum. Statistical methods and outcome measures: Hemodynamic measurements and histological studies. MAIN FINDINGS: All uterine myometrial patches survived in the test animals. Fluoroscopic hemodynamic measurements were made for ejection fractions at 8 weeks after the application of the uterine patch. Histological study demonstrated well-healed myometrial-myocardium junctions with minimum scar tissue. Angiogenesis occurred in the transplanted myometrium. Connexin 43 expression was demonstrated in the transplanted patches. CONCLUSION: Our noncontrolled preliminary rabbit experiments indicate that patches of uterine myometrium reinforced by greater omentum can be used as autologous transplant therapy for infracted myocardium. This is an innovative technique that could lead to future treatment for individuals who may suffer from an infarcted myocardium and may not be eligible for traditional reperfusion therapy. PMID- 18286457 TI - Electrospun bioresorbable heart valve scaffold for tissue engineering. AB - Currently marketed mechanical or biological prosthetic heart valves are regarded as valid substitutes for native heart valves suffering from degenerative pathologies. These devices require strict follow-up due to dysfunctions or post surgical complications. Potential drawbacks of these medical devices are calcification, tearing of the cusps, thromboembolism and hemolysis. In this context, a tissue engineering approach offers a promising alternative scenario. In this paper, a trileaflet poly(epsilon-caprolactone) (PCL) heart valve scaffold prototype has been manufactured by electrospinning technique using a custom-made rotating target. Process parameters were selected in order to achieve suitable microstructure and mechanical performance. The electrospun heart valve prototype was functionally characterized by means of a pulse duplicator in order to evaluate the mechanical/hydraulic response to the imposed testing conditions. Leaflets synchronously opened in the ejection phase and the proper apposition of the leaflets prevented high leakage volumes in the diastolic phase. This preliminary study suggests a successful perspective for the proposed approach in designing a novel tissue engineered bioresorbable heart valve. PMID- 18286458 TI - Neurophysiological assessment of the electrostimulation procedures used in stroke patients during rehabilitation. AB - The aim of this study was to evaluate the effectiveness of the associated electrotherapeutical and kinesiotherapeutical treatment in patients after ischemic stroke (N=24), mainly by means of neurophysiological tests. All patients underwent the same 20 days of neurorehabilitation procedures. Particular attention was paid to three-stage modified electrotherapy procedures such as: oververtebral functional electrical stimulation (FES), transcutaneous electrical nerve stimulation (TENS) and the alternate neuromuscular functional electrical stimulation (NMFES) of antagonistic muscles of the wrist and the ankle (N=16). Electrotherapy was supplemented with kinesiotherapeutic (mainly PNF) procedures acting as an amplifier. Clinical assessment included muscle tension (Ashworth's scale), muscle force (Lovett's scale) and reflex scoring at wrist and ankle. However, the effectiveness of the procedures was measured by the assessment of results in complex and repetitive, bilaterally performed global electromyography (EMG) and electroneurography (ENG; M-wave studies). The statistical analysis obtained from results in clinical and neurophysiological examinations suggested that the dorsiflexion of wrist and ankle was improved in the majority of patients who took part in this study. EMG and ENG examinations showed that 20 days of therapy improved both activity in muscle motor units on the more paralyzed side (mainly within upper extremities) and to a lesser degree in the transmission of efferent impulses within motor fibers of nerves. The results obtained suggest that patients after ischemic strokes never show an isolated unilateral disability in motor functions. No definite similarities between the results of clinical and neurophysiological studies were found, which may suggest greater accuracy of the neurophysiological evaluation. PMID- 18286459 TI - Psychometric properties of the Chinese version of the Swanson, Nolan, and Pelham, version IV scale - parent form. AB - This study aimed to establish the psychometric properties of parent ratings on the Chinese version of the Swanson, Nolan, and Pelham IV scale (SNAP-IV) in a school-based sample of 3534 students in grades 1 to 8 from two cities and two suburbs in Taiwan and 189 children diagnosed with attention deficit/hyperactivity disorder (ADHD) (aged 6 to 15) consecutively recruited from a medical center in Taipei. Parents completed the Chinese versions of the SNAP-IV, Strengths and Difficulties Questionnaire, and Child Behavior Checklist. The Chinese SNAP-IV demonstrated similar three factor structure (Inattention, Hyperactivity/Impulsivity, and Oppositional) as its English version, and satisfactory test-retest reliability (intraclass correlation = 0.59 approximately 0.72), internal consistency (alpha = 0.88 approximately 0.90), concurrent validity (Pearson correlations = 0.56 approximately 0.72), and discriminant validity. Boys scored higher than girls across the eight school grade levels. The SNAP-IV clearly distinguished children with ADHD from school-based participants. Comorbidity with oppositional defiant disorder/conduct disorder predicted higher SNAP-IV scores among children with ADHD. Our findings suggest that the Chinese SNAP-IV is a reliable and valid instrument for rating ADHD-related symptoms in both clinical and community settings in Taiwan. PMID- 18286460 TI - Psychometric characteristics of Cloninger's criteria for personality disorder in a population of French prisoners. AB - Cloninger has proposed a new procedure to assess personality disorders on the basis of his model of temperament and character. We explored the psychometric characteristics of this instrument in 560 male prisoners, using factor analyses. Results showed that the 20 items relating to the general personality criteria have acceptable internal consistency and that the three-factor structure of the temperament items used to subtype personality disorder are correctly identified by a maximum likelihood factor analysis model with varimax rotation. Overall, 32% of prisoners had a personality disorder according to this model, and it was possible to subtype 68% of these. This new diagnostic procedure for personality disorder seemed to have satisfactory basic psychometric characteristics but further study is required to explore the concurrent validity and the reliability of this instrument. PMID- 18286461 TI - Ascertaining late-life depressive symptoms in Europe: an evaluation of the survey version of the EURO-D scale in 10 nations. The SHARE project. AB - The reported prevalence of late-life depressive symptoms varies widely between studies, a finding that might be attributed to cultural as well as methodological factors. The EURO-D scale was developed to allow valid comparison of prevalence and risk associations between European countries. This study used Confirmatory Factor Analysis (CFA) and Rasch models to assess whether the goal of measurement invariance had been achieved; using EURO-D scale data collected in 10 European countries as part of the Survey of Health, Ageing and Retirement in Europe (SHARE) (n = 22,777). The results suggested a two-factor solution (Affective Suffering and Motivation) after Principal Component Analysis (PCA) in 9 of the 10 countries. With CFA, in all countries, the two-factor solution had better overall goodness-of-fit than the one-factor solution. However, only the Affective Suffering subscale was equivalent across countries, while the Motivation subscale was not. The Rasch model indicated that the EURO-D was a hierarchical scale. While the calibration pattern was similar across countries, between countries agreement in item calibrations was stronger for the items loading on the affective suffering than the motivation factor. In conclusion, there is evidence to support the EURO-D as either a uni-dimensional or bi-dimensional scale measure of depressive symptoms in late-life across European countries. The Affective Suffering sub-component had more robust cross-cultural validity than the Motivation sub-component. PMID- 18286462 TI - Exploring daily variations of drinking in the Swiss general population. A growth curve analysis. AB - This study aims to address the underlying trajectories of weekly individual drinking patterns by growth models and to relate differences in drinking patterns to socio-demographic and drinking characteristics of respondents. Data came from a two-stage stratified random subsample of 747 persons aged 15 years or more from a Swiss study on alcohol consumption using a within-subject design conducted between March 1999 and July 1999. Beverage specific assessment of daily alcohol consumption was obtained by a weekly drinking diary and other characteristics via telephone interviews. The diary had to be filled out on seven consecutive days. The growth models accounted for up to 37.6% of the initial error variance and provided evidence for two distinct, negatively correlated underlying trajectories of drinking patterns. The first trajectory described an increase in consumption from Monday to Sunday. The second trajectory was about a specific weekend consumption culminating on Saturday with a significantly higher growth rate among young people and heavy episodic drinkers than in other subgroups. Therefore, young and heavy episodic drinkers may be exposed to sudden adverse consequences of alcohol consumption during the weekend. Prevention efforts which are targeted to this subgroup should take its specific drinking pattern into account. PMID- 18286463 TI - Mapping a multidimensional emotion in response to television commercials. AB - Unlike previous emotional studies using functional neuroimaging that have focused on either locating discrete emotions in the brain or linking emotional response to an external behavior, this study investigated brain regions in order to validate a three-dimensional construct--namely pleasure, arousal, and dominance (PAD) of emotion induced by marketing communication. Emotional responses to five television commercials were measured with Advertisement Self-Assessment Manikins (AdSAM) for PAD and with functional magnetic resonance imaging (fMRI) to identify corresponding patterns of brain activation. We found significant differences in the AdSAM scores on the pleasure and arousal rating scales among the stimuli. Using the AdSAM response as a model for the fMRI image analysis, we showed bilateral activations in the inferior frontal gyri and middle temporal gyri associated with the difference on the pleasure dimension, and activations in the right superior temporal gyrus and right middle frontal gyrus associated with the difference on the arousal dimension. These findings suggest a dimensional approach of constructing emotional changes in the brain and provide a better understanding of human behavior in response to advertising stimuli. PMID- 18286464 TI - Meta-analyses of the incidence and prevalence of schizophrenia: conceptual and methodological issues. AB - While meta-analytic techniques are routine in the synthesis of data from randomized controlled trials, there are no clear guidelines on how best to summarize frequency data such as incidence and prevalence estimates. Based on data from two recent systematic reviews of the incidence and prevalence of schizophrenia, this paper explores some of the conceptual and methodological issues related to the meta-analyses of frequency estimates in epidemiology. Because variations in the incidence and prevalence of disorders such as schizophrenia can be informative, there is a case against collapsing data into one pooled estimate. Variations in frequency estimates can be displayed graphically, or summarized with quantiles around measures of central tendency. If pooled estimated are of interest, then researchers need to be aware that studies based on large samples will leverage greater weight on the pooled value. Based on systematic reviews of the incidence and prevalence of schizophrenia, we explore if these and related issues are of practical concern. When used with appropriate caution, meta-analysis can complement the synthesis of frequency data in epidemiology; however, researchers interested in variation should not rely on meta-analysis alone. PMID- 18286465 TI - Psychometric properties of the Chinese version of the adult ADHD Self-report Scale. AB - Although attention deficit/hyperactivity disorder (ADHD) in adulthood has become a topic of considerable interest to psychiatrists over the past decade, little is known about the prevalence or correlates of adult ADHD in the Chinese population. As a first step in addressing this problem, this study presents data on the psychometric properties of the Chinese version of the World Health Organization's Adult ADHD Self-report Scale (ASRS) in a sample of 1031 young males from an army base and 3298 young adults from two colleges. All participants completed the Chinese ASRS. Participants from an army base also completed a clinical assessment including the Wender Utah Rating Scale (WURS) for assessing childhood ADHD, the Impulsiveness Scale for measuring the severity of current impulsive behaviors, and information about substance use. Results showed good concordance (intraclass correlations = 0.80 approximately 0.85) and internal consistency (Cronbach's alpha = 0.83 approximately 0.91) among the ASRS subscales and moderate to high correlations between these subscales and the WURS (Pearson's correlations = 0.37 approximately 0.66). The Chinese ASRS also demonstrated the ability to predict childhood disruptive problems and habitual use of substances. Our findings suggest that the Chinese ASRS is a reliable and valid instrument to assist in screening for adult ADHD. PMID- 18286466 TI - Malignant Peripheral Nerve Sheath Tumor: molecular pathogenesis and current management considerations. AB - Malignant Peripheral Nerve Sheath Tumors (MPNSTs) are rare tumors that often occur in patients with neurofibromatosis 1. Surgical resection represents the mainstay of treatment. Radiation and chemotherapy have a role in selected patients with MPNST. Accurate pathologic diagnosis remains a challenge in many cases of MPNST. There are many recent advances in the understanding of the molecular pathogenesis of MPNST which represent the best opportunities to develop new strategies for management of patients with MPNST. PMID- 18286467 TI - Inhibition of Cdc7/Dbf4 kinase activity affects specific phosphorylation sites on MCM2 in cancer cells. AB - The Cdc7/Dbf4 kinase is required for initiation of DNA replication and also plays a role in checkpoint function in response to replication stress. Exactly how Cdc7/Dbf4 mediates those activities remains to be elucidated. Cdc7/Dbf4 physically interacts with and phosphorylates the minichromosome maintenance complex (MCM), such as MCM2, MCM4 and MCM6. Cdc7/Dbf4 activity is required for association of Cdc45 followed by recruitment of DNA polymerase on the chromatin. Using high resolution mass spectrometry, we identified six phosphorylation sites on MCM2, two of them have not been described before. We provide evidence that Cdc7/Dbf4 mediates phosphorylation on serine 108 and serine 40 on human MCM2 in vitro and in vivo in cancer cells in the absence of DNA damage. Antibodies specific to pS108 or pS40 confirmed the sites and established useful read-outs for inhibition of Cdc7/Dbf4. This report demonstrates the utility of an in vitro to in vivo workflow utilizing immunoprecipitation and mass spectrometry to map phosphorylation sites on endogenous kinase substrates. The approach can be readily generalized to identify target modulation read-outs for other potential kinase cancer targets. PMID- 18286468 TI - Gene expression profiling of macrophages following mice treatment with an immunomodulator medication. AB - Canova (CA) is a complex homeopathic medication used in diseases where the immune system is depressed. Previous studies demonstrated that it is neither toxic nor mutagenic and activates macrophages. We now evaluate CA effects on cytokine production and gene expression from mice macrophages. The global view of changes in expression of genes with known functions can provide a vivid picture of the way in which cell adapts to a changing environment or a challenge. We found a decrease in IL-2 and IL-4 production and a differential expression in 147 genes from CA group. These genes are mainly involved in transcription/translation, cell structure and dynamics, immune response, cytoprotection, enzymatic process, and receptors/ligands. With gene expression analysis we state that this medication provokes a reaction that involves alterations in gene expression profile mainly in the ones involved with macrophages activation, corroborating the laboratorial research and the clinical data. PMID- 18286469 TI - MAPRes: an efficient method to analyze protein sequence around post-translational modification sites. AB - Functional switches are often regulated by dynamic protein modifications. Assessing protein functions, in vivo, and their functional switches remains still a great challenge in this age of development. An alternative methodology based on in silico procedures may facilitate assessing the multifunctionality of proteins and, in addition, allow predicting functions of those proteins that exhibit their functionality through transitory modifications. Extensive research is ongoing to predict the sequence of protein modification sites and analyze their dynamic nature. This study reports the analysis performed on phosphorylation, Phospho.ELM (version 3.0) and glycosylation, OGlycBase (version 6.0) data for mining association patterns utilizing a newly developed algorithm, MAPRes. This method, MAPRes (Mining Association Patterns among preferred amino acid residues in the vicinity of amino acids targeted for post-translational modifications), is based on mining association among significantly preferred amino acids of neighboring sequence environment and modification sites themselves. Association patterns arrived at by association pattern/rule mining were in significant conformity with the results of different approaches. However, attempts to analyze substrate sequence environment of phosphorylation sites catalyzed for Tyr kinases and the sequence data for O-GlcNAc modification were not successful, due to the limited data available. Using the MAPRes algorithm for developing an association among PTM site with its vicinal amino acids is a valid method with many potential uses: this is indeed the first method ever to apply the association pattern mining technique to protein post-translational modification data. PMID- 18286470 TI - A panel of ancestry informative markers for estimating individual biogeographical ancestry and admixture from four continents: utility and applications. AB - Autosomal ancestry informative markers (AIMs) are useful for inferring individual biogeographical ancestry (I-BGA) and admixture. Ancestry estimates obtained from Y and mtDNA are useful for reconstructing population expansions and migrations in our recent past but individual genomic admixture estimates are useful to test for association of admixture with phenotypes, as covariate in association studies to control for stratification and, in forensics, to estimate certain overt phenotypes from ancestry. We have developed a panel of 176 autosomal AIMs that can effectively distinguish I-BGA and admixture proportions from four continental ancestral populations: Europeans, West Africans, Indigenous Americans, and East Asians. We present allele frequencies for these AIMs in all four ancestral populations and use them to assess the global apportionment of I-BGA and admixture diversity among some extant populations. We observed patterns of apportionment similar to those described previously using sex and autosomal markers, such as European admixture for African Americans (14.3%) and Mexicans (43.2%), European (65.5%) and East Asian affiliation (27%) for South Asians, and low levels of African admixture (2.8-10.8%) mirroring the distribution of Y E3b haplogroups among various Eurasian populations. Using simulation studies and pedigree analysis we show that I-BGA estimates obtained using this panel and a four-population model has a high degree of precision (average root mean square error [RMSE]=0.026). Using ancestry-phenotype associations we demonstrate that a large and informative AIM panel such as this can help reduce false-positive and false-negative associations between phenotypes and admixture proportions, which may result when using a smaller panel of less informative AIMs. PMID- 18286471 TI - What we have learned and will learn from cell ultrastructure in embedment-free section electron microscopy. AB - The limitations inherent in conventional electron microscopy (EM) using epoxy ultrathin sections for a clear recognition of biological entities having electron densities similar to or lower than that of epoxy resin have led to the development of embedment-free sectioning for EM. Embedment-free section EM is reliably performed using water-soluble polyethylene glycol (PEG) as a transient embedding medium, with subsequent de-embedment of PEG by immersion into water, followed by critical point-drying (CPD) of the embedment-free section. The present author has stressed that this approach clearly discloses structures whose contours and/or appearance are accordingly vague and/or fuzzy in conventional EM, but does not reveal any new structures. Based on embedment-free electron microscopy (PEG-EM), this article presents five major findings regarding strand- or microtrabecular lattices which have been clearly revealed to occur in the cytoplasmic matrix-an impossibility with conventional EM. These are (1) the appearance of lattices of different compactness in various cells and in intracellular domains of a given cell; (2) the faithful reproduction from an albumin solution in vitro of strand-lattices with correspondingly increasing compactness following increasing concentrations; (3) the appearance of more compact lattices from gelated gelatin than from solated gelatin at a given concentration in vitro; (4) the appearance of either greater or less lattice compactness by hyper- or hypotonic pretreatments of cells; and (5) the appearance of certain intracellular proteins confined to the centripetal demilune-domain of centrifuged ganglion cells which is occupied with strand-lattices of a substantial compactness. From these findings, questions now arise as to the biological significance of the individual strand itself in the microtrabecular lattices in PEG-EM. In addition, it may be that the appearance of strand-lattices in a given biological domain represents the presence of soluble proteins; the lattice-compactness indicates the concentration of soluble proteins in the domain, and the aqueous cytoplasm is equivalent to the aqueous solution. Further, the appearance of two contiguous lattice domains exhibiting differing degrees of compactness in a given cell indicates that cytoplasmic proteins are solated in a domain with less compact lattices, whereas they are gelated in the other domain. These proposed interpretations need to be confirmed by further studies. If confirmed, the control mechanisms of the localization and movement of intracellular organelles could then be understood on the basis not only of information about the cytoskeletons but also of cell ultrastructure-related information on the concentration and sol-gel states of intracellular proteins. In addition, possible interpretations of the significance of strand-lattices in PEG EM are also applicable to the nucleoplasm, especially extra-heterochromatin (euchromatin) areas. Finally, several potential uses/advantages of PEG-EM in the cell-ultrastructure have also been demonstrated, especially in three-dimensional reconstructions of nonmembranous structures including stereo-viewing using a pair of EM images with appropriate tilting as well as electron microscopic tomography. PMID- 18286472 TI - A tribute to Murray F. Brennan, MD. PMID- 18286473 TI - Diagnosis and management of lipomatous tumors. AB - Lipomatous tumors range from benign lipomas to high-grade liposarcomas. Liposarcomas are classified into five histologic subtypes: well-differentiated, dedifferentiated, myxoid, round cell, and pleomorphic, which differ in outcomes and patterns of recurrence. Surgical resection is the mainstay of curative treatment; however, large, high grade liposarcomas may benefit from multimodality treatment with chemotherapy and radiation. A histologic-subtype specific nomogram provides accurate survival predictions. Prospective randomized clinical trials will continue to improve our care of patients with liposarcoma. PMID- 18286474 TI - Diagnosis and management of synovial sarcoma. AB - Synovial sarcoma accounts about 9% of soft tissue sarcomas, most commonly develops in the extremity of young adults, is considered high grade and contains a characteristic translocation (X;18;p11;q11). While surgery and radiation therapy have achieved excellent local control, distant metastasis remains the principal problem limiting survival. Although ifosfamide based chemotherapy has been associated with an improved survival in patients with synovial sarcoma, the search for less toxic and more targeted systemic therapies is ongoing. PMID- 18286475 TI - Malignant tumors of blood vessels: angiosarcomas, hemangioendotheliomas, and hemangioperictyomas. AB - Sarcomas that arise from or resemble the components of blood vessels are uncommon and include angiosarcomas, hemangioendotheliomas, and hemangiopericytomas. This article reviews the management of these three types of sarcomas. Diagnosis can sometimes be difficult, with the diagnosis of hemangiopericytoma versus solitary fibrous tumor currently in debate. Each of these sarcomas subtypes has certain unique clinical traits. The mainstay of treatment is surgical resection, sometimes combined with radiation therapy. Adjuvant chemotherapy is unproven but can be considered. For patients with advanced disease, various chemotherapeutic regimens may result in meaningful responses in a minority of patients. PMID- 18286476 TI - Diagnosis and management of pleomorphic sarcomas (so-called "MFH") in adults. AB - The existence of "malignant fibrous histiocytoma" ("MFH") as a distinct entity is controversial. Previously accepted as the most common sarcoma affecting adults, it is now known to comprise a heterogeneous group of tumors without a specific known line of differentiation. Reclassification of many tumors in this group afforded better prognostication, but traditional treatments still apply. Pleomorphic soft tissue tumors for which a line of differentiation is debatable are presently categorized as undifferentiated pleomorphic sarcoma. PMID- 18286477 TI - Gastrointestinal stromal tumors and leiomyosarcomas. AB - Only recently has gastrointestinal stromal tumor (GIST) been recognized as the most frequent GI mesenchymal neoplasm. Prior to the use of KIT staining, most GI stromal tumors were identified as leiomyosarcoma (LMS). For primary GIST, surgery remains the principal treatment and adjuvant imatinib may improve outcomes. Multimodality therapy may also be effective in patients with metastatic GIST. In this review, we summarize the epidemiology, clinicopathologic features, natural history, and clinical management of LMS and GIST. PMID- 18286478 TI - Pediatric sarcomas occurring in adults. AB - Sarcomas arise in any part of the body, in any age group. Some sarcomas have a particular predilection for the pediatric population, and often bear specific chromosomal translocations. These "pediatric" sarcomas still occur in adults, often times with higher risk of dying of disease compared to children with a comparable diagnosis. The management of some of these rare tumors is discussed herein, including osteogenic sarcoma, Ewing sarcoma, and rhabdomyosarcoma. PMID- 18286479 TI - Grb10/Nedd4-mediated multiubiquitination of the insulin-like growth factor receptor regulates receptor internalization. AB - The adaptor protein Grb10 is an interacting partner of the IGF-I receptor (IGF IR) and the insulin receptor (IR). Previous work from our laboratory has established the role of Grb10 as a negative regulator of IGF-IR-dependent cell proliferation. We have shown that Grb10 binds the E3 ubiquitin ligase Nedd4 and promotes IGF-I-stimulated ubiquitination, internalization, and degradation of the IGF-IR, thereby giving rise to long-term attenuation of signaling. Recent biochemical evidence suggests that tyrosine-kinase receptors (RTK) may not be polyubiquitinated but monoubiquitinated at multiple sites (multiubiquitinated). However, the type of ubiquitination of the IGF-IR is still not defined. Here we show that the Grb10/Nedd4 complex upon ligand stimulation mediates multiubiquitination of the IGF-IR, which is required for receptor internalization. Moreover, Nedd4 by promoting IGF-IR ubiquitination and internalization contributes with Grb10 to negatively regulate IGF-IR-dependent cell proliferation. We also demonstrate that the IGF-IR is internalized through clathrin-dependent and-independent pathways. Grb10 and Nedd4 remain associated with the IGF-IR in early endosomes and caveosomes, where they may participate in sorting internalized receptors. Grb10 and Nedd4, unlike the IGF-IR, which is targeted for lysosomal degradation are not degraded and likely directed into recycling endosomes. These results indicate that Grb10 and Nedd4 play a critical role in mediating IGF-IR down-regulation by promoting ligand-dependent multiubiquitination of the IGF-IR, which is required for receptor internalization and regulates mitogenesis. PMID- 18286480 TI - Additional MDA-MB-231 breast cancer cell matrix metalloproteinases promote invasiveness. AB - We are interested in two aspects of a given type of metastatic breast cancer: which potentially cancer-relevant genes are expressed and which factors determine invasiveness. Using reverse transcription real-time PCR, we detected gene expression of 26 matrix metalloproteinases (MMPs) in MDA-MB-231 breast cancer cells, including those of MMP-12, MMP-16 variant 2, MMP-19, MMP-20, MMP-21, MMP 23, MMP-24, MMP-25, MMP-25 variant 2, MMP-L1, MMP-26, MMP-27, and MMP-28, in contrast to the 13 MMPs detected until now in these cells. We found that MMP genes are expressed at widely different levels in these cells, over five orders of magnitude. After individual siRNA-induced depletions, we found that six additional species of cancer cell MMPs promote invasiveness in MDA-MB-231 cells: MMP-3, MMP-11, MMP-12, MMP-17, MMP-19, and MMP-23, thus raising the total to 12 endogenous MMPs which do so in these cells. The data support the conclusion that some cancer cell MMPs, although expressed at low levels, are needed for cancer trait in MDA-MB-231 cells, and that several endogenous MMPs play non-redundant roles in this process. The mRNA level of MMP-11, but not of other MMPs, rose substantially following individual siRNA-targeted depletion of cancer cell MMP-17 mRNA, while no MMP mRNA increased appreciably after degradation of other MMP mRNAs. This supports the conclusion that MMP-17 may be a member of an intracellular signaling pathway which downregulates MMP-11 mRNA. PMID- 18286481 TI - The 20th Aspen Cancer Conference: Mechanisms of Toxicity, Carcinogenesis, Cancer Prevention, and Cancer Therapy 2005. PMID- 18286482 TI - Combined BubR1 protein down-regulation and RASSF1A hypermethylation in Wilms tumors with diverse cytogenetic changes. AB - BUB1B and RASSF1A genes play specific roles in the mitotic checkpoint, and their defects may cause chromosome instability or aneuploidy in mouse fibroblasts and human cancer cell lines; however, few studies have reported a correlation between defects in these genes and chromosome changes in human tumor samples. We examined chromosome abnormalities in 25 Wilms tumors by metaphase comparative genomic hybridization, and classified them into 14 hyperdiploid (50 > or = chromosomes), 2 near-or-pseudodiploid, and 9 diploid tumors. We also examined various molecular aspects of BUB1B and RASSF1A, and evaluated the relationship between chromosome changes and the status of both genes. No tumors showed BUB1B mutation. BubR1 protein (BUB1B gene product) expression was undetectable or decreased in five of six hyperdiploid or near-or-pseudodiploid tumors and increased in four of five diploid tumors, whereas all seven tumors examined showed BUB1B mRNA expression irrespective of their chromosome pattern. Furthermore, while complete promoter methylation of RASSF1A was found in 13 of 16 hyperdiploid or near-or pseudodiploid tumors, unmethylated RASSF1A was found in 5 of 9 diploid tumors. Partial RASSF1A methylation was found in three hyperdiploid or near-or pseudodiploid tumors and in four diploid tumors. Thus, BubR1 protein expression decreased, and the promoter region of RASSF1A was completely methylated in the great majority of hyperdiploid or near-or-pseudodiploid tumors, BubR1 protein expression increased and RASSF1A was unmethylated in the majority of diploid tumors. These findings suggest that the combined BubR1 protein down-regulation and RASSF1A hypermethylation might be implicated in the formation of chromosomal changes found in Wilms tumors. PMID- 18286483 TI - Resveratrol and X rays affect gap junction intercellular communications in human glioblastoma cells. AB - Resveratrol (3,4',5-trihydroxystilbene) is a polyphenol synthesized by a wide variety of plant species in response to injury, UV irradiation and fungal attack. Many studies have revealed a variety of resveratrol intracellular targets whose modulation gives rise to overlapping responses leading to growth arrest and death. Many authors have reported different human cancer cell lines, treated with resveratrol at micromolar concentrations, arrested their proliferative cycle in the G1/S boundary or in the S phase and this cell cycle arrest was followed by apoptotic death. Less is known about the ability of resveratrol to modify the effect of radiation exposure in normal and cancer cells. Considering that controlled exposure to ionizing radiation is one of the most used treatments in cancer patients and that these schedules are not always effective in medical practice, as in the case of glioma patients, the testing of combined treatment protocols (resveratrol and ionizing radiation) could be of interest, opening the door to future studies which would examine the pharmacological activity of resveratrol. In this study we have looked into whether resveratrol is able to modulate cell cycle progression in human glioblastoma cells and to regulate GJs expression in cancer cells. With this aim in mind we have performed a cytofluorimetric multiparameter assay to quantify the presence of GJs in U87 glioma cells treated with resveratrol and/or X rays. We report that resveratrol induces a delay in cell cycle progression and both alone and in combination with X rays is able to enhance gap junction Intercellular Communications. PMID- 18286484 TI - Pilot study: volatile organic compounds as a diagnostic marker for head and neck tumors. AB - BACKGROUND: In the last decade, the analysis of volatile organic compounds (VOC) has undergone a rapid development. In this pilot study, patients with HNSCC were tested with a proton transfer reaction-mass spectrometry in order to establish a minimal invasive screening method. METHODS: Overall in a period of 2 years, 22 carcinoma patients were recruited for the study. All patients had a newly diagnosed histologically secured squamous cell carcinoma of the upper aerodigestive tract. These results were statistically compared with 3 control groups: healthy controls, high-risk, and posttherapy patients. RESULTS: Two hundred nine different masses were measured; 188 of these were evaluated. The statistical workup of the 4 study groups produced 42 different masses, which showed a statistically significant difference from the carcinoma group compared with the control groups. CONCLUSION: A screening method for HNSCC using VOC seems to be possible, but further investigation is necessary. PMID- 18286485 TI - Alveolar soft part sarcoma of the larynx: a case report of an unusual location with immunohistochemical and ultrastructural analyses. AB - BACKGROUND: Alveolar soft part sarcoma (ASPS) is a rare mesenchymal neoplasm of uncertain origin. In this article, we report a case of ASPS occurring in the larynx, an extremely rare location for this rather unusual tumor. METHODS AND RESULTS: The patient was a 34-year-old Japanese woman who requested an examination for hoarseness. The tumor showed a proliferation of large polygonal cells with periodic-acid-Schiff-positive diastase-resistant intracytoplasmic granules, arranged in an alveolar growth pattern. The cytoplasm of the tumor cells was eosinophilic. Tumor cells were positive for vimentin and titin. Nuclear immunoreactivity for TFE3 was observed, and the Ki-67 labeling index was 14.7%. Ultrastructurally, electron-dense rod-shaped crystals were infrequently observed in the cytoplasm. This case was finally diagnosed as ASPS of the larynx. CONCLUSION: We discuss the histogenesis and differential diagnosis of ASPS with immunohistochemical and ultrastructural findings. TFE3 immunohistochemistry was found to be a very useful marker for the diagnosis of ASPS. PMID- 18286486 TI - Oral primary angiosarcoma of the lower lip mucosa: report of a case in a 15-year old boy. AB - BACKGROUND: Angiosarcomas are rare soft tissue malignant tumors with dismal prognosis. Head and neck involvement is uncommon (5%) and usually affects the scalp or facial skin. METHODS AND RESULTS: We present the case of an inferior lip mucosal low-grade angiosarcoma in a 15-year-old boy treated exclusively with surgery. One and a half years after treatment, the patient was free of signs of recurrence. Prompt and accurate diagnosis with adequate imaging modalities and multidisciplinary treatment are crucial for optimal management of these neoplasms. CONCLUSION: Lip mucosal involvement is exceptional with only a few cases described in the literature, all in patients older than 60 years To our knowledge, this is the youngest patient ever reported. PMID- 18286487 TI - Lateral oromandibular defect: when is it appropriate to use a bridging reconstruction plate combined with a soft tissue revascularized flap? AB - BACKGROUND: A quasi-experimental retrospective study was undertaken to evaluate a new concept of free tissue volume restoration combined with bridging reconstruction plate (compartment approach) to reduce plate-related complication rates. METHODS: We evaluated 40 patients with large lateral mandible defects and associated complex soft tissue defects reconstructed with a revascularized soft tissue flap and titanium hollow screw reconstruction plates. A case-control comparison was performed based on reconstruction type: restoration of soft tissue defect (conventional approach-group 1) versus over-reconstruction of soft tissue defect (compartment approach-group 2). RESULTS: Plate exposure rate was 6 of 16 (38%) in group 1 versus 2 of 24 (8%) in group 2, and the difference was statistically significant (p = .04). The mean time to exposure was 10 months. Plate fracture rate was 6 of 23 (26.1%) in dentulous patients versus 1 of 17 (5.9%) in edentulous patients. Gastrostomy tube dependence was 6 of 16 (38%) in group 1 versus 6 of 24 (25%) in group 2. CONCLUSION: The "compartment approach" reduces plate exposure rate and gastrostomy tube dependence. Revascularized osseocutaneous reconstruction is still required in dentulous patients. PMID- 18286488 TI - N2-N3 neck nodal control without planned neck dissection for clinical/radiologic complete responders-results of Trans Tasman Radiation Oncology Group Study 98.02. AB - BACKGROUND: The aim of this study was to determine the incidence of isolated nodal failure in patients with N2/3 disease who achieved a complete clinical and radiological response (CR) at 12 weeks postchemoradiation, when no planned neck dissection was performed. METHODS: We analyzed the nodal response and subsequent neck control of 102 patients with initial N2/3 disease treated on the Trans Tasman Radiation Oncology Group 98.02 study. RESULTS: With a median 4.3 years follow-up, the patterns of first failure in the CR patients were local 4%, local and nodal 2%, distant 28%, and locoregional plus distant (within 1 month) 6%. There were no patients who had only neck failure. CONCLUSION: Patients in this trial with N2/3 disease who obtained a clinical and radiological complete response to chemoradiation had a zero incidence of isolated neck failure without a planned neck dissection. The continued use of planned neck dissections in this patient subset cannot be justified. PMID- 18286489 TI - Vidian nerve schwannoma with middle cranial fossa extension resected via a maxillary swing approach. AB - BACKGROUND: Vidian nerve schwannoma is an extremely rare type of facial nerve schwannoma. To the best of our knowledge, only 1 case has been reported. METHODS: We report an additional case of vidian nerve schwannoma with middle cranial fossa extension in a 49-year-old Japanese woman. The surgical approaches for infratemporal fossa schwannomas are reviewed, and the maxillary swing approach we used is described. RESULTS: We adopted a maxillary swing approach combined with endonasal endoscopic techniques for the resection of the lesion. Gross total resection was achieved without sacrificing the trigeminal nerves or the facial motor nerves. The postoperative course was uneventful. CONCLUSION: Vidian nerve schwannoma is located in the retro-maxillary space. It can grow silently until it involves the median skull base extensively. The maxillary swing approach was useful in this case. PMID- 18286490 TI - Horizontal supraglottic partial laryngectomy for selected squamous carcinoma of the vallecula. AB - BACKGROUND: Our aim was to determine the incidence of local control in patients with selected squamous carcinoma of the vallecula treated with horizontal supraglottic laryngectomy; to analyze the consequences of local recurrence in terms of nodal recurrence, distant metastasis, survival, causes of death, overall local control, and laryngeal preservation; and to identify any clinical factors predictive of these outcomes. METHODS: This was a retrospective nonrandomized case series in a university teaching hospital. An inception cohort of 95 previously untreated patients were followed until death or for a minimum of 5 years. According to the 2002 Union Internationale Contre le Cancer (UICC) staging classification system, the tumor was classified as T1, T2, and T3 in 13, 60, and 22 patients, respectively, while disease in 67 patients was considered to be in stages III to IV. All patients underwent a horizontal partial supraglottic partial laryngectomy. Ninety-four patients had an associated neck dissection. An induction chemotherapy regimen was used in 91 patients; postoperative radiation therapy was given for 49 patients. The main outcome measures were local recurrence, nodal recurrence, distant metastasis, and survival. RESULTS: The 1-, 3-, and the 5-year actuarial survival estimates were 86.3%, 64.2%, and 47.4%, respectively. Overall, the main causes of death were as follows: metachronous second primary tumor (47.2% of patients), intercurrent disease (16.7%), distant metastasis (15.3%), local recurrence (6.3%), and nodal recurrence, (4.2%). The 1 , 3-, and 5-year actuarial local recurrence rates were 4.5%, 11%, and 11%, respectively. Nine patients developed a local recurrence; 3 were successfully salvaged. Using multivariate analysis, no single variable was found to increase the risk for local recurrence. The overall laryngeal preservation rate and local control rate were 89.5% (85/95) and 93.4% (89/95), respectively. Local recurrence was associated with a significant increase in nodal recurrence (p <.04) and distant metastasis (p = .03). CONCLUSIONS: Based on this experience, horizontal partial supraglottic laryngectomy appears to be a valid approach for functional organ-preservation in patients with selected T1-T3 SCC of the vallecula. PMID- 18286491 TI - Using fluorodeoxythymidine to monitor anti-EGFR inhibitor therapy in squamous cell carcinoma xenografts. AB - BACKGROUND: 3'-18F-fluoro-3'-deoxy-fluorothymidine (18F-FLT), a nucleoside analog, could monitor effects of molecularly targeted therapeutics on tumor proliferation. METHODS: We tested whether (18)F-FLT positron emission tomography (PET) uptake changes are associated with antitumor effects of erlotinib in A431 xenografts or cetuximab in SCC1 xenografts. RESULTS: Compared with pretreatment FLT PET scans, 3 days of erlotinib in A431 reduced the standardized uptake value (SUV) by 18%, whereas placebo increased SUV by 1% (p = .005). One week of cetuximab in SCC1 reduced SUV by 62%, whereas placebo reduced SUV by 16% (p = .005). FLT uptake suppression following anti-epidermal growth factor receptor (EGFR) treatment was associated with reduced tumor thymidine kinase-1 (TK1) activity. In vitro TK1 knockdown studies confirmed the importance of TK1 activity on intracellular FLT accumulation suppression. CONCLUSIONS: 18F-FLT PET imaging detects tumor responses to EGFR-inhibitors within days of starting therapy. This technique may identify patients likely to benefit from EGFR-inhibitors early in their treatment course. PMID- 18286492 TI - Results of endoscopic resection followed by radiotherapy for primarily diagnosed adenocarcinomas of the paranasal sinuses. AB - BACKGROUND: Adenocarcinoma is the most frequent histological subtype of paranasal sinus malignancy diagnosed in Belgium. Classical treatment consists of an external surgical approach (lateral rhinotomy with medial maxillectomy or craniofacial resection) followed by radiotherapy. The role, possibilities, and limitations of endoscopic sinus surgery (ESS) are to date unknown. METHODS: We studied 44 patients with primary (not treated previously) adenocarcinoma treated with endoscopic sinus surgery and radiotherapy between 1992 and 2004. RESULTS: The median follow-up of the patients alive at the end of the study period was 36 months. For the 3-year follow-up, the overall survival, disease-specific survival, and local control rate were 81%, 91%, and 73%, respectively. Corresponding rates for the 5-year follow-up were 53%, 83%, and 62%. Union Internationale Contre le Cancer T classification did not appear to influence these results. CONCLUSIONS: Endoscopic sinus surgery followed by radiotherapy for primary adenocarcinoma of the paranasal sinuses gives oncological results comparable to those of standard external approaches. PMID- 18286493 TI - Outcomes following reirradiation of patients with head and neck cancer. AB - BACKGROUND: This study reports the outcomes for patients with head and neck cancer who received reirradiation with palliative or curative intent. METHODS: A retrospective review of 41 patients treated with curative (n = 28) or palliative (n = 13) reirradiation was conducted. Survival was calculated from the start of the reirradiation. Radiation-related toxicities were classified according to Radiation Therapy Oncology Group criteria. Disease-related problems included adverse events during or after reirradiation that were not directly related to reirradiation. RESULTS: The observed 1-year survival for all patients was 39.0% (23.0% palliative, 46.3% curative). Median survival for all patients was 10.2 months. Seventy-five percent of curative and 53.8% of palliative patients had grade 3 or 4 radiation-related toxicities and/or major disease-related problems. CONCLUSION: A second course of radiotherapy in patients with head and neck cancer should be offered with a clear understanding that survival is poor and many of these patients will suffer severe radiation-related or disease-related insults to their quality of life during and after treatment. PMID- 18286494 TI - Composite hemangioendothelioma of the oral cavity: case report and review of the literature. AB - BACKGROUND: Composite hemangioendothelioma is a rare histopathologic feature, which is part of the hemangioendothelioma family. This is a heterogeneous group of vascular neoplasia with a high tendency to local relapse but a rare predisposition to metastatic spread. Composite hemangioendothelioma mainly affects adults and is usually localized in the distal extremity of the limbs. To date, only 1 case has been detected in the oral cavity. METHODS: A case of composite hemangioendothelioma in a 38-year-old man is reported. The lesion was localized in the left cheek vestibular mucosa. It was surgically excised with a 1 cm safety margin. Reconstruction was performed with a platysma myocutaneous flap. RESULTS: At present, the patient is free from recurrence. The aesthetic and functional outcomes are satisfactory. CONCLUSION: This case demonstrates how a composite hemangioendothelioma histological diagnosis can be difficult to achieve. Furthermore, therapy must be surgical and excision should be wide. PMID- 18286495 TI - Exclusive use of acid citrate dextrose for anticoagulation during extracorporeal photopheresis in patients with contraindications to heparin: an effective protocol. AB - Extracorporeal photopheresis (ECP) routinely uses heparin for anticoagulation. For patients with contraindications to heparin, alternative anticoagulation using acid citrate dextrose (ACD-A) has been reported to be safe and effective. However, detailed ECP protocols that exclusively use ACD-A anticoagulation and minimize citrate toxicity have not yet been published. We report a protocol that completely replaces heparin with ACD-A for ECP, which was developed at the University of California, Los Angeles (UCLA), and our experience since its implementation. ECP was performed with the UVAR XTS photopheresis system using ACD-A and control of the rate of citrate infusion. Calcium gluconate solution was administered prophylactically and as needed for symptoms of citrate toxicity. The medical records of patients who underwent ECP using the ACD-A protocol between January 2003 and July 2006 were reviewed. The incidence and severity of citrate toxicity and the technical data for all procedures were analyzed. During this period, 94 ECP procedures were performed with ACD-A anticoagulation on five patients. All patients tolerated the procedures well without significant complications. Only minimal symptoms of citrate toxicity (grade 1) were observed in 24.5% of all procedures; symptoms resolved promptly following administration of additional calcium gluconate. In conclusion, an effective protocol for ECP using ACD-A anticoagulation exclusively in patients with contraindications to heparin employs continuous monitoring of flow rates and prophylactic administration of calcium gluconate to minimize citrate toxicity. PMID- 18286496 TI - Cause of mortality in five-year survivors of childhood cancer in North-West Italy, 1967-1999. PMID- 18286497 TI - Intraoperative radiation therapy as an "early boost" in locally advanced head and neck cancer: preliminary results of a feasibility study. AB - BACKGROUND: The acute toxicity of intraoperative radiation therapy (IORT) delivered as an "early boost" after tumor resection in patients with locally advanced head and neck cancer was evaluated. METHODS: Twenty-five patients were enrolled in the study. All patients underwent surgery with radical intent, and 17 had microvascular flap reconstruction. The IORT was delivered in the operating room. Twenty patients received adjuvant external beam radiation therapy (EBRT). RESULTS: Five patients experienced various degrees of complications in the postoperative period, all of which were treated conservatively. One patient had a partial flap necrosis after EBRT that was treated with flap removal. Six deaths were recorded during the mean follow-up period of 8 months; none of the deaths were related to radiation treatment. CONCLUSION: This feasibility study shows that the use of IORT as an early boost is feasible with no increase in acute toxicity directly attributable to radiation. PMID- 18286498 TI - Right common carotid artery crossing the midline neck anterior to the trachea: a cadaver case report. AB - BACKGROUND: The course of the proximal common carotid artery is reasonably uniform. Its normal position is taken for granted when performing invasive procedures, but variations should be in back of the mind of the surgeon. METHODS: This is a case report from an anatomical dissection of the neck and mediastinum of a cadaver. RESULTS: The right common carotid artery was the first branch of the aorta and ascended on the left side, anterolateral to the trachea. It crossed the midline anterior to the trachea between the sternal notch and the isthmus of the thyroid gland. Once on the right side, the artery resumed a normal course. A retro-esophageal right subclavian artery was also found. CONCLUSION: Anomalous variations in anatomy do occur, the possibilities of which the surgeon needs to be aware. This case illustrates a previously unreported anatomical anomaly that would clearly have had catastrophic consequences had this not been noted during surgery on the anterior neck. PMID- 18286499 TI - Use and application of mode of action in cancer risk assessment. PMID- 18286500 TI - Defibrotide in sinusoidal obstruction syndrome: mounting evidence in pediatric patients (Commentary on Qureshi et al., page 831). PMID- 18286501 TI - Treatment of children with metastatic soft tissue sarcoma with oral maintenance compared to high dose chemotherapy: report of the HD CWS-96 trial. AB - PURPOSE: We prospectively studied the efficacy of high dose therapy (HDT) versus an oral maintenance treatment (OMT) in patients with stage IV soft tissue sarcoma (STS). PATIENTS AND METHODS: Both groups were pretreated with the CEVAIE combination consisting of carboplatin, etoposide, vincristine, actinomycin D, ifosfamide, and epirubicin. HDT consisted of a tandem cycle of thiotepa (600 mg/m(2)) plus cyclophosphamide (4,500 mg/m(2)) and melphalan (120 mg/m(2)) plus etoposide (1,800 mg/m(2)). This treatment was compared with OMT, consisting of four cycles trofosfamide (10 days 2 x 75 mg/m(2)/day) plus etoposide (10 days 2 x 25 mg/m(2)/day), and 4 cycles trofosfamide (10 days 2 x 75 mg/m(2)/day) plus idarubicin (10 days 4 x 5 mg/m(2)). Eligibility criteria were: diagnosis confirmed by reference pathology, primary stage IV, below 22 years of age, and having completed the study therapy. RESULTS: From 96 patients 45 were treated with HDT and 51 with OMT. The main risk parameters were equally distributed in both arms. After a median follow-up of 57.4 months, 11/45 (24.4%) patients in the HDT-arm and 26/51 (57.8%) patients in OMT-arm were alive. Kaplan-Meier analysis demonstrated an overall survival for the whole group of 0.27 (OMT group: 0.52, HDT group 0.27, log rank P = 0.03). The proportional hazard analysis for patients with rhabdomyosarcoma (RMS) or "RMS-like" tumors (77.1% of all patients) demonstrated an independent benefit of OMT on outcome. CONCLUSION: Oral maintenance therapy seems to be a promising option for patients with RMS-like stage IV tumors. PMID- 18286502 TI - Defibrotide in the prevention and treatment of veno-occlusive disease in autologous and allogeneic stem cell transplantation in children. AB - BACKGROUND: Hepatic veno-occlusive disease (VOD) is a common (10-50%) and serious complication of haematological stem cell transplantation (HSCT), with up to 90% mortality rates. We carried out a study to assess whether the use of prophylactic defibrotide in paediatric patients undergoing HSCT results in a lower frequency or severity of hepatic VOD. PROCEDURE: Forty-seven successive patients who underwent transplantation between April 2004 and December 2005 were given defibrotide prophylaxis and were compared with 56 historical controls transplanted between November 2001 and April 2004. No serious side effects were reported. High risk patients in the control group received ursodeoxycholic acid and tinzaparin as VOD prophylaxis. The groups were matched for sex, age, type of transplant and risk. RESULTS: In the defibrotide group, four patients developed clinical VOD (Seattle criteria) although two had liver biopsies which showed graft versus host disease (GvHD). Defibrotide dose was increased and symptoms resolved within 14 days. Of the control group four patients had VOD. Two of these patients had reversed hepatic vein flow and died 30 days post-transplant, partly due to VOD. VOD was associated with busulfan conditioning (P = 0.001) and not with age, sex, type of transplant, GvHD, abnormal liver function prior to transplant or type of antifungal prophylaxis. CONCLUSIONS: VOD incidence and severity was reduced in the defibrotide group which suggests that defibrotide might be effective in preventing and treating VOD. Sufficiently powered randomised trials are now required to definitively test the role of defibrotide in this setting. PMID- 18286503 TI - Intravascular ultrasound-guided central vein angioplasty and stenting without the use of radiographic contrast agents. AB - Patients with contraindications to iodinated radiographic contrast agents present a significant challenge during endovascular intervention. A 46-year-old man with end-stage renal disease and a normally functioning left upper extremity arteriovenous fistula presented with severe left arm edema. The patient's history included repeated severe anaphylactoid reactions with severe respiratory distress upon exposure to iodinated contrast. In an attempt to avoid the use of iodinated contrast, angioplasty and stent placement of a severe central venous stenosis were performed using only fluoroscopy and intravascular sonography. In patients unable to receive iodinated contrast secondary to anaphylactoid reactions, intravascular sonography can be used to guide angioplasty and stenting of central venous stenosis. PMID- 18286504 TI - Leiomyoma of the testis: a rare testicular mass. AB - We report the sonographic appearance of a testicular leiomyoma, which is a rare smooth muscle tumor of the testis. The patient presented with a painless testicular mass that was confirmed as an intratesticular tumor on sonographic examination. Histopathologic examination revealed a testicular leiomyoma. PMID- 18286505 TI - Cancer vaccines: clinical development challenges and proposed regulatory approaches for patient access to promising treatments. AB - The challenges of late-stage development increasingly are becoming clear as the clinical development of therapeutic cancer vaccines continues to progress. Preclinical and clinical research had indicated that cancer vaccines exert optimal benefit in earlier stage disease or in adjuvant/minimal residual disease (MRD) settings. However, clinical trials in these settings can be prohibitively slow from a development perspective because of the better prognosis of the patient population and the current lack of early surrogate markers of efficacy. Therefore, the 'optimal' patient population (the patient group in which the greatest benefit is demonstrated) for any given vaccine studied in this setting generally must be identified first through the conduct of a large randomized trial for each indication, then confirmed in a second large randomized trial. On the basis of the current regulatory paradigm, a late-stage development program for a cancer vaccine in the earlier stage disease or adjuvant/MRD setting easily could extend past 10 years. The tight funding environment and constant evolution in medical practice, which can make replication of results from the first trial infeasible over such a long timeline, pose additional challenges. In this report, the authors discuss 3 potential regulatory solutions to better enable the development and commercialization of therapeutic cancer vaccines: a U.S. Food and Drug Administration-proposed cost-recovery program, conditional marketing authorization, and a new development paradigm. All of these solutions aim to balance a complex equation of biologic rationale, weight of the evidence for efficacy and safety, regulatory expectations, and cost and timeline of clinical development. PMID- 18286506 TI - In-hospital complications of autologous hematopoietic stem cell transplantation for lymphoid malignancies: clinical and economic outcomes from the Nationwide Inpatient Sample. AB - BACKGROUND: Autologous hematopoietic stem cell transplantation (auto HSCT) is standard of care therapy for multiple myeloma and Hodgkin and non-Hodgkin lymphomas in front-line and salvage settings, respectively. Complications remain common, but population-based estimates of their frequency and relative contribution to cost are not available. METHODS: A retrospective cohort comprised of 8891 patients with multiple myeloma and lymphoma admitted to US hospitals for auto HSCT over a 2-year period (2000-2001) was extracted from the Nationwide Inpatient Sample (NIS). Patient characteristics, vital status, and total hospital charges were obtained directly from the NIS. Transplant characteristics and outcomes were identified by ICD-9-CM codes. Mean hospital charges were examined by outcome and transformed into cost by using Medicare cost-to-charge ratios. Factors associated with hospital cost, length of stay, and in-hospital mortality were explored by using multivariate regression. RESULTS: The mean hospital cost for auto HSCT during this period was $51,312. Significant complications were documented for >50% of admissions. Infectious complications (~60%) and stomatitis (~40%) were the most frequent, and both were associated with increased hospital costs (range, $15,000 to $50,000). In-hospital mortality was rare (<5%) but was associated with markedly increased cost when it occurred. Pretransplant conditioning with total body irradiation was strongly associated with infectious complications, higher cost, and death. CONCLUSIONS: Adverse events are both common and costly after auto HSCT. Strategies to minimize complications could significantly reduce not only morbidity and mortality but also the cost of the procedure. Administrative data can be profitably exploited to investigate outcomes in this population. PMID- 18286507 TI - Pneumonectomy after chemoradiation: the Dana-Farber Cancer Institute/Brigham and Women's Hospital experience. AB - BACKGROUND: The current study was conducted to examine the outcomes of pneumonectomy after induction chemoradiotherapy in patients with locally advanced nonsmall cell lung cancer (NSCLC). METHODS: All patients undergoing pneumonectomy after induction therapy at the Brigham and Women's Hospital were retrospectively evaluated for 30-day and 100-day mortality and treatment-related complications with Institutional Review Board approval. Multivariate and univariate analyses for clinical factors correlating with toxicity and/or survival were calculated. RESULTS: Between 1995 and 2005, 73 patients underwent pneumonectomy for NSCLC after induction therapy. All patients received radiation (median dose of 54 gray [Gy]) and 69 patients (95%) received concurrent chemotherapy. The median age was 62 years and 43 patients (59%) were male; Thirty-seven patients (51%) had American Joint Committee on Cancer stage IIIA NSCLC, 27 (37%) had stage IIIB, 6 had stage IIB, and 4 had stage IV NSCLC because of a resected solitary brain metastasis. A majority (44; 60%) of patients received the combination of carboplatin and paclitaxel, whereas 15 (21%) received the combination of cisplatin and etoposide. Forty-five patients (62%) underwent left pneumonectomy. With a median follow-up of 28 months, the 1-year and 2-year overall survival rates were 70% and 49%, respectively. The 30-day and 100-day mortality rates were 6% and 10%, respectively. Only 4 of 73 patients (6%) died of acute respiratory distress syndrome. The rate of nonfatal treatment-related morbidity was 11%. On univariate analysis, right-sided pneumonectomy was associated with a higher risk of treatment-related mortality (P = .099). CONCLUSIONS: With an acceptable mortality rate, a single-institutional series demonstrated that trimodality therapy including pneumonectomy can be safely accomplished in patients with advanced NSCLC. PMID- 18286508 TI - p38/NF-kB-dependent expression of COX-2 during differentiation and inflammatory response of chondrocytes. AB - Studying cartilage differentiation, we observed the emergence of inflammation related proteins suggesting that a common pathway was activated in cartilage differentiation and inflammation. In the present paper, we investigated the expression pathway of the inflammation-related enzyme Cyclooxygenase-2 (COX-2) during differentiation and inflammatory response of the chondrocytic cell line MC615. Cells were cultured either as (i) proliferating prechondrogenic cells expressing type I collagen or (ii) differentiated hyperconfluent cells expressing Sox9 and type II collagen. The p38 and the NF-kB pathways were investigated in standard conditions and after inflammatory agents treatment. NF-kB was constitutively activated in differentiated cells. The activation level of NF-kB in differentiated cells was comparable to the level in proliferating cells treated with the inflammatory agent LPS. In both cases, p65 was bound to the NF kB consensus sequence of COX-2 promoter. p38, constitutively activated in differentiated cells, was activated in proliferating cells by treatment with LPS or IL-1alpha. In stimulated proliferating cells the two pathways are connected since addition of the p38-specific inhibitor SB203580 inhibited p38 activation, significantly reduced NF-kB activation and repressed COX-2 synthesis indicating that p38 is upstream NF-kB activation and COX-2 synthesis. In differentiated cells, the treatment with the inflammatory agent neither enhance NF-kB activation, nor synthesis of COX-2 while the addition of SB203580 neither repressed activation of p38, nor COX-2 synthesis, suggesting a constitutive activation of a p38/NF-kB/COX2 pathway. Our data indicate that in chondrocytes, COX-2 is expressed via p38 activation/NF-kB recruitment during both differentiation and inflammatory response. PMID- 18286509 TI - Short- and long-term A3 adenosine receptor activation inhibits the Na+/H+ exchanger NHE3 activity and expression in opossum kidney cells. AB - The renal function of the A(3) adenosine receptor (A3AR) is poorly characterized. In this study, we report that the A3AR-selective agonist, 1-[2-chloro-6-[[(3 iodophenyl)methyl]amino]-9H-purine-9-yl]-1-deoxy-N-methyl-b-D-ribofuranuronamide (2-Cl-IBMECA) regulates the Na+/H+ exchanger-3 (NHE3) in a dose- and time dependent fashion. In opossum kidney (OK) cells, 2-Cl-IBMECA at high (10(-6) M) and low (10(-8) M) dose inhibits NHE3 by a multiphasic time course with an acute phase of NHE3 inhibition from 15 min to 1 h, followed by a chronic phase of NHE3 inhibition from 24 to 48 h. Pre-incubation with either the selective A3AR antagonist MRS1523 (10(-7) M) or the protein kinase C inhibitor, Calphostin C (10(-8) M) completely blocked 10(-6) M 2-Cl-IBMECA-induced acute (15 min) and chronic (24 h) phases of NHE3 inhibition. In contrast, the acute inhibitory phase (15 min) of 10(-8) M 2-Cl-IBMECA was completely prevented only when Calphostin C (10(-8) M) was added in conjunction with the protein kinase A inhibitor, H89 (10( 7) M). Acute (15 or 30 min depending on the A3AR-agonist concentration) A3AR dependent inhibition of NHE3 activity was accompanied by decrease in cell surface NHE3 protein with no change in total NHE3 antigen. Chronic (24 h) A3AR-mediated down-regulation of NHE3 was associated with reduction of surface NHE3, decreased total NHE3 protein (70%) and a paradoxical rise of NHE3 RNA (40%). In summary, these results indicate that A3AR directly regulates NHE3 at multiple levels in a complex pattern. A3AR-dependent short- and long-term inhibition of NHE3 may be a fundamental mechanism of net sodium and fluid balance. PMID- 18286510 TI - Somatosensory system in two types of motor reorganization in congenital hemiparesis: topography and function. AB - This study investigates the (re-)organization of somatosensory functions following early brain lesions. Using functional magnetic resonance imaging (fMRI), passive hand movement was studied. Transcranial magnetic stimulation (TMS) and magnetoencephalography (MEG) were used as complementary methods. fMRI data was analyzed on the first level with regard to topographical variability; second-level group effects as well as the overall integrity of the somatosensory circuitry were also assessed. Subjects with unilateral brain lesions occurring in the third trimester of pregnancy or perinatally with different types of motor reorganization were included: patients with regular, contralateral motor organization following middle cerebral artery strokes (CONTRA(MCA), n = 6) and patients with reorganized, ipsilateral motor functions due to periventricular lesions (IPSI(PL), n = 8). Motor impairment was similar, but sensory impairment was more pronounced in the CONTRA(MCA) group. Using fMRI and MEG, both groups showed a normal pattern with a contralateral somatosensory representation, despite the transhemispherically reorganized primary motor cortex in the IPSI(PL) group, as verified by TMS. Activation topography for the paretic hands was more variable than for the nonparetic hand in both groups. The cortico-cerebellar circuitry was well-preserved in almost all subjects. We conclude that in both models of motor reorganization, no interhemispheric reorganization of somatosensory functions occurred. Also, no relevant intrahemispheric reorganization was observed apart from a higher topographical variability of fMRI activations. This preserved pattern of somatosensory organization argues in favor of a differential lesion effect on motor and somatosensory functions and demonstrates a limited compensatory potential for the latter. PMID- 18286511 TI - Safety and efficacy of arsenic trioxide for patients with advanced metastatic melanoma. AB - BACKGROUND: Arsenic trioxide (ATO) cytotoxicity and apoptosis induction has been demonstrated with numerous cancer cell lines, including human melanoma. METHODS: A second-line, phase 2, single-arm study of ATO was conducted in patients with inoperable American Joint Committee on Cancer (AJCC) stage IV melanoma. One cycle consisted of a loading dose of 0.32 mg/kg/day for 4 days in Week 1, followed by 0.25 mg/kg/day twice per week for 6 weeks, followed by 1 week of rest, at which time response assessment was performed. RESULTS: Twenty-one patients (median age, 63.8 years) were accrued. All had stage IV melanoma including M1a (2 patients), M1b (6 patients), and M1c (13 patients) disease. One patient had metastatic choroidal melanoma and 20 patients had cutaneous melanoma. Twenty patients had received prior therapy. Possible treatment-related grade 3 of 4 toxicities (using the National Cancer Institute Common Toxicity Criteria) included 1 case of idiopathic thrombocytopenic purpura and 1 case of elevated lactate dehydrogenase. Four patients did not complete the first cycle of therapy and were not evaluable for response. Among 17 evaluable patients, 1 patient (6%; 95% confidence interval [95% CI], 0-29%) achieved a partial response lasting 7 months, and 10 patients (59%) had disease stabilization after at least 1 cycle, but all eventually developed disease progression. The median time to disease progression was 17 weeks (95% CI, 11-38 weeks) and the median survival was 13 months (95% CI, 12-26 months). CONCLUSIONS: ATO as tested in the current trial was found to be well tolerated and had limited activity in patients with metastatic melanoma. The application of this agent in combination with either chemotherapy or agents that target recognized critical signaling and antiapoptotic pathways of melanoma has not yet been performed. PMID- 18286512 TI - Is there an optimal comorbidity index for prostate cancer? AB - BACKGROUND: Comorbidity is an important consideration in oncology practice, particularly among older patients. Although a variety of comorbidity indices have been employed in research studies, it is unclear whether any one index is preferred. METHODS: An age-stratified random sample of 345 men (mean age of 69 years) who were newly diagnosed with prostate cancer were identified from a cancer registry in Ontario, Canada. Comorbidity and treatment information were obtained from chart review. Four comorbidity indices were utilized: Charlson Index, Diagnosis Count, Index of Coexistent Disease (ICED), and number of medications. Logistic regression analysis was used to compare the performance of comorbidity measures with respect to predicting receipt of curative treatment (radical prostatectomy or radiotherapy) and overall 6-year survival. Multivariable model performance including each of the comorbidity measures was compared by calculating the area under the receiver operating characteristic curve (AUROC). RESULTS: Among men with localized disease (n = 231), in models adjusted for age, Gleason score, and prostate-specific antigen level, only the Charlson Index was found to be a statistically significant predictor of receipt of curative treatment (P < .05), although all comorbidity indices had similar AUROC in adjusted models. After a median follow-up of 6.5 years, 116 of 345 men (33.6%) had died. In adjusted models, all 4 comorbidity indices performed similarly in predicting overall survival. CONCLUSIONS: Although comorbidity is an important predictor of both curative treatment and overall survival in prostate cancer, the optimal comorbidity index for use in research remains unclear. Selecting the optimal comorbidity index may depend on both the specific patient population and the outcome being considered. PMID- 18286513 TI - Delayed intervention of sporadic renal masses undergoing active surveillance. AB - BACKGROUND: Prompt surgical management remains the standard of care for renal cell carcinoma (RCC). Occasionally, it is necessary to postpone or delay surgical treatment. The authors of this report assessed whether delayed intervention following a period of active surveillance altered minimally invasive or nephron sparing treatment plans, increased the risk of stage progression, and/or decreased recurrence-free survival rates. METHODS: The authors searched their institutional kidney cancer database to identify small (< or =4 cm in greatest dimension on presentation), enhancing renal masses for which treatment initially was delayed or refused. Clinical, radiographic, and pathologic records were reviewed to determine linear tumor growth kinetics, alterations in treatment plan, stage migration, and cancer-specific outcomes related to delayed intervention. RESULTS: Eighty-seven sporadic, localized, enhancing renal masses were identified in 82 patients who had management postponed for a median of 14 months (mean, 21 months; range, 6-97 months). Median tumor diameter was 2.0 cm on presentation. Treatment in 60 of 87 tumors (69%) was delayed for > or =12 months, and treatment was delayed for > or =24 months in 29 of 87 tumors (33%). Overall, 66 of 87 tumors (76%) underwent nephron-sparing approaches. In addition, 52 of 87 tumors (60%) were treated in a minimally invasive fashion. Pathology confirmed RCC in 73 of 87 treated tumors (84%). Fourteen of 54 tumors (26%) that were treated by surgical extirpation were high-risk tumors, and 3 of 54 tumors (6%) were upstaged on pathologic review. CONCLUSIONS: The majority of small, sporadic, clinically localized renal tumors demonstrated slow interval growth. The management of these lesions may be delayed cautiously without limiting or complicating the available treatment options or incurring a high risk of disease progression. PMID- 18286514 TI - Contrast-enhanced sonographic characteristics of neovascularization in carotid atherosclerotic plaques. AB - PURPOSE: To evaluate neovascularization within carotid atherosclerotic plaques with contrast-enhanced sonography. METHODS: We used contrast-enhanced sonography to examine 63 patients with carotid atherosclerotic plaques. The features of neovascularization within the plaques were analyzed and correlated with plaque size and echogenicity. RESULTS: There were 81 atherosclerotic plaques, 62 of which (43 soft and 19 mixed) enhanced after injection of a contrast agent. The enhancement occurred from the carotid wall to the center of the plaque with a short-line pattern in 36 plaques, whereas 26 plaques enhanced from both the carotid wall and the carotid lumen, with a sparse spot pattern. The arrival time of contrast was shorter (p < 0.001) and time to peak was longer (p < 0.001) in the plaques than in the carotid lumen. Time to peak was shorter, whereas enhanced intensity was greater in soft plaques than in mixed plaques (p < 0.01 and p < 0.05, respectively). Among the 19 unenhanced plaques, 6 were hard, 3 were calcified, 3 were soft, and 7 were mixed. The thickness of the unenhanced plaques was <2.4 mm. CONCLUSION: Contrast-enhanced sonography allows the noninvasive, dynamic evaluation of neovascularization within carotid plaques, and the presence of neovascularization may correlate with plaque morphology. PMID- 18286515 TI - Role of power Doppler sonography in early diagnosis of osteomyelitis in children. AB - We describe 2 children complaining of leg pain associated with elevated C reactive protein levels in whom power Doppler (PD) sonography suggested the diagnosis of early osteomyelitis. PD sonography detected increased blood flow that resulted in a high-intensity signal area adjacent to the symptomatic tibia surface. Antibiotic therapy led to prompt improvement. These cases suggest that PD sonography is useful in the evaluation of possible early osteomyelitis in children. PMID- 18286516 TI - Nomograms of fetal thyroid measurements estimated by 2-dimensional sonography. AB - PURPOSE: To build nomograms of fetal thyroid circumference (FTC), fetal thyroid area (FTA), and fetal thyroid transverse diameter (FTTD) throughout gestational age (GA). METHOD: Between January 2006 and July 2006, FTC, FTA, and FTTD were measured once in 196 normal fetuses examined at a GA of 22-35 weeks. Inclusion criteria were a healthy mother with normal maternal thyrotropin level during pregnancy, a singleton pregnancy with normal fetal morphology on sonography, and GA confirmed via first-trimester sonographic examination. RESULTS: Mean FTC, FTA, and FTTD ranged from 3.21 cm, 0.58 cm(2), and 1.19 cm at 22 weeks to 5.11 cm, 1.69 cm(2), and 1.89 cm at 35 weeks, respectively. Linear regression analysis yielded the following formulas for FTC, FTA, and FTTD according to GA: FTC (cm) = 0.146 x GA (weeks); FTA (cm(2)) = -1.289 + 0.085 x GA (weeks); FTTD (cm) = 0.054 x GA (weeks). The following logarithmic formulas were obtained for the expected fetal thyroid measurements according to estimated fetal weight (FW): FTC (cm) = 4.791 + 1.265 x logN FW; FTA (cm(2)) = -1.676 + 0.455 x logN FW; and FTTD (cm) = 0.399 + 0.001 x logN FW. CONCLUSION: We describe new nomograms of fetal thyroid measurements throughout gestation that may be useful in case of thyroid dysfunction. PMID- 18286517 TI - Mammary duct ectasia in infant breast with bloody nipple discharge: sonographic findings. AB - Bloody nipple discharge is very rare in childhood. We report the sonographic findings of mammary duct ectasia and cystic changes under the nipple with abnormal content involving a 3-month-old boy with bloody nipple discharge. PMID- 18286518 TI - Comment on "Transcranial Doppler ultrasonographic evaluation of cerebral circulation during passive tilting in patients with Parkinson's disease". PMID- 18286519 TI - Mobile phone electromagnetic radiation activates MAPK signaling and regulates viability in Drosophila. AB - Mobile phones are widely used in the modern world. However, biological effects of electromagnetic radiation produced by mobile phones are largely unknown. In this report, we show biological effects of the mobile phone 835 MHz electromagnetic field (EMF) in the Drosophila model system. When flies were exposed to the specific absorption rate (SAR) 1.6 W/kg, which is the proposed exposure limit by the American National Standards Institute (ANSI), more than 90% of the flies were viable even after the 30 h exposure. However, in the SAR 4.0 W/kg strong EMF exposure, viability dropped from the 12 h exposure. These EMF exposures triggered stress response and increased the production of reactive oxygen species. The EMF exposures also activated extracellular signal regulated kinase (ERK) and c-Jun N terminal kinase (JNK) signaling, but not p38 kinase signaling. Interestingly, SAR 1.6 W/kg activated mainly ERK signaling and expression of an anti-apoptotic gene, whereas SAR 4.0 W/kg strongly activated JNK signaling and expression of apoptotic genes. In addition, SAR 4.0 W/kg amplified the number of apoptotic cells in the fly brain. These findings demonstrate that the exposure limit on electromagnetic radiation proposed by ANSI triggered ERK-survival signaling but the strong electromagnetic radiation activated JNK-apoptotic signaling in Drosophila. PMID- 18286520 TI - Pulsed electromagnetic fields induced femoral metaphyseal bone thickness changes in the rat. AB - The effect of 1 Hz, 30 mT pulsed magnetic fields on young adult rat femoral metaphyseal bone thickness was assessed. Ten same litter, female Wistar rats were studied; five of them underwent 30 min magnetic stimulation sessions for 20 consecutive days. The anterior and posterior cortical, as well as trabecular bone transverse thicknesses were measured. The results obtained under clear field microscopy in stimulated and control histological cuts were (in microm) 398 +/- 32 versus 260 +/- 22 (P = 0.002), 380 +/- 68 versus 252 +/- 21 (P = 0.03), and 168 +/- 11 versus 112 +/- 11 (P = 0.002), respectively. The transcranial magnetic stimulation system, approved for human therapy, generates pulsed electromagnetic fields, which induce a significant thickness increase in cortical and trabecular in vivo stimulated bone tissues. This is the first time this effect in healthy animals is shown. PMID- 18286521 TI - Sarcoma of the pulmonary trunk and the main pulmonary arteries. AB - We report on a sarcoma of the central pulmonary arteries. Surgical therapy consisted in replacing both main pulmonary arteries and the pulmonary trunk including the pulmonary valve. Six months later a left-sided pneumonectomy had to be performed due to an intravascular tumor. Fifteen months after first resection treatment, recurrent tumors of the right pulmonary artery and the right ventricle were resected. Two years after the first operation the patient has no detectable tumor. PMID- 18286522 TI - Carcinomas arising in the submandibular gland: high propensity for systemic failure. AB - BACKGROUND: Cancers of the submandibular gland are uncommon and only a few small series have reported patient survival and prognosis. METHODS: We examined the treatment outcomes of 62 patients with surgically treated submandibular gland carcinomas. All patients underwent surgical excision with/without neck dissection, and 41 received postoperative radiotherapy for high-grade, invasive, positive margin, or regionally metastatic tumors. The locoregional control and survival rates were calculated by the Kaplan-Meier method and prognostic factors were calculated from uni- and multivariate analyses. RESULTS: Of the 62 submandibular gland carcinomas, 19 were adenoid cystic, 11 were mucoepidermoid, and 10 were salivary duct carcinomas, and 8 were carcinomas in pleomorphic adenoma. Actuarial 5-year locoregional control, distant metastasis-free survival, disease-free and overall survival rates were 69.7%, 65.8%, 52.8%, and 56.8%, respectively. In multivariate analysis, T category and histological grading were prognostic for disease-free survival (P < 0.01), and T category and resection margins were prognostic for locoregional control (P < 0.02). Distant metastases were found in 21 patients (33.9%) at initial staging (n = 2) or follow-up (n = 19). CONCLUSIONS: Despite effective locoregional treatment, approximately one third of patients with submandibular gland carcinomas may fail systemically, resulting in poor survival. New, more effective therapies may be required for these patients. PMID- 18286523 TI - Expression of P53, P27 and KI-67 in colorectal cancer patients of various ethnic origins: clinical and tissue microarray based analysis. AB - BACKGROUND: This study was conducted to determine survival according to the expression of molecular markers in colorectal cancer (CRC) patients of various ethnic origins. METHODS: Resection of primary tumor was conducted on 171 patients with CRC. Corresponding archived paraffin-embedded blocks were retrieved and tissue microarray (TMA) constructed. Immunohistochemical staining of the TMA for p53, p27 and Ki-67 was quantified by two independent pathologists. Survival was analyzed using the Kaplan-Meier product limit method. RESULTS: With a median follow-up of 65 months, 56 patients (32.7%) died of disease. AJCC stage correlated with disease-free (DFS, P < 0.0001) and overall survival (OS, P < 0.0001). IHC staining was positive for Ki-67 in 77.4%, p53 in 55.8% and p27 in 54.2% of patients. Primary tumor marker expression did not correlate with DFS or OS. The 5-year DFS for Ashkenazi Jews was 75%, significantly higher than Sephardic Jews (SJ) 64% and Palestinian Arabs (PA) 38%, P = 0.001. CONCLUSIONS: Ethnicity among Ashkenazi and SJ and PA appears to have a significant impact on disease outcome in patients with CRC patients, while primary tumor expression of p53, p27 and Ki-67 was unrelated to disease outcome. PMID- 18286524 TI - Effect of season on thermoregulation, metabolism and ventilation of the southern brown bandicoot Isoodon obesulus (Marsupialia: Peramelidae). AB - Seasonal variation in various thermoregulatory, metabolic and ventilatory parameters was examined for southern brown bandicoots (Isoodon obesulus fusciventer) from a Mediterranean climate near Perth, Western Australia. There was significant seasonal variation over the four annual seasons at thermoneutrality (Ta=30 degrees C) in body temperature, oxygen consumption, carbon dioxide production, respiratory exchange ratio, total evaporative water loss, wet and dry thermal conductance and tidal volume but not mass, ventilatory frequency, minute volume or oxygen extraction efficiency. Only carbon dioxide production and respiratory exchange ratio showed an annual pattern that was significantly related to season, with both being significantly higher in winter, presumably as a result of greater and higher quality food availability. PMID- 18286525 TI - Natural killer cell neoplasms. AB - Natural killer (NK) cell tumors are an uncommon and heterogeneous group of disorders. The World Health Organization (WHO) classified mature NK cell neoplasms into 2 types: 1) extranodal NK cell lymphoma, nasal type and 2) aggressive NK cell leukemia. The mature NK cell tumors are prevalent in Asia and Central and South America. These tumors show polymorphic neoplastic infiltrate with angioinvasion and/or angiodestruction, cytoplasmic azurophilic granules, CD2 positive (CD2+)/CD3-negative (CD3-)/cCD3epsilon+/CD56+ phenotype, and strong association with Epstein-Barr virus (EBV). Loss of chromosomes 6q, 11q, 13q, and 17p are recurrent aberrations. Although blastic NK cell lymphoma, currently referred to as CD4+/CD56+ hematodermic neoplasm, also was included in the NK cell lymphoma category in the WHO classification scheme, existing evidence indicates a plasmacytoid dendritic cell derivation as opposed to an NK cell origin. Recently, rare cases of CD56+ immature lymphoid tumors have been reported in the literature. These tumors are characterized by blastic appearance, CD3-/CD4 /CD56+/CD13-/CD33- phenotype, T-cell receptor and immunoglobulin genes in germline configuration, and no evidence of EBV, suggesting a true immature NK cell derivation. For this article, the authors reviewed the recent concepts and progress in clinicopathologic features, pathogenesis, genetic characteristics, diagnosis, differential diagnosis, treatment approaches, and outcomes of all subtypes of NK cell neoplasms. PMID- 18286526 TI - Risk of early recurrence among postmenopausal women with estrogen receptor positive early breast cancer treated with adjuvant tamoxifen. AB - BACKGROUND: Adjuvant aromatase inhibitors (AIs), instead of or after tamoxifen, are effective in decreasing recurrence in postmenopausal women with estrogen receptor (ER)-positive breast cancer. An understanding of which patients are at risk of early recurrence while they are receiving tamoxifen may improve clinical decision making. METHODS: The patients who were included in this study were women aged >or= 50 years with early-stage, ER-positive breast cancer diagnosed between 1986 and 1999 and had been treated with tamoxifen. Characteristics of the patients with early recurrences (within 2.5 years of diagnosis), late recurrences (between 2.5 years and 5 years) and no recurrence within 5 years were compared. Logistic regression analyses were conducted to identify which groups were at risk of early recurrence. RESULTS: Among 3844 women, 304 women (7.9%) developed disease recurrence within 2.5 years. Higher than average rates of recurrence within 2.5 years were observed in cohorts with lymph node (N)-positive tumors (11.5%), grade 3 histology (14.3%), or low-positive ER levels, ie, 10-49 fmol/mg or 10%-20% staining (14.9%). In multivariate analyses, only pathologically N positive tumors (1-3 vs 0 positive lymph nodes: odds ratio [OR], 1.6; 4-9 vs 0 positive lymph nodes: OR, 2.23 [P= .03]) and low-positive ER status (OR, 2.04; P= .01) were associated with recurrence within 2.5 years compared with recurrence between 2.5 years and 5 years. Other clinical and pathologic variables were not predictive of early recurrence. CONCLUSIONS: Subgroups of women with early ER positive breast cancer may be identified who are at increased risk of recurrence within 2.5 years of diagnosis despite tamoxifen. It remains to be proven whether upfront AI therapy results in an advantage to these women. PMID- 18286527 TI - Matched case-control phase 2 study to evaluate the use of a frozen sock to prevent docetaxel-induced onycholysis and cutaneous toxicity of the foot. AB - BACKGROUND: Onycholysis occurs in approximately 30% of patients treated with docetaxel. The efficacy and safety of an Elasto-Gel frozen sock (FS) was investigated for the prevention of docetaxel-induced nail and skin toxicity of the feet. METHODS: Patients receiving docetaxel at a dose of 70 to 100 mg/m(2) every 3 weeks were eligible for this matched case-control study. Each patient wore an FS for 90 minutes on the right foot. The unprotected left foot acted as control. Nail and skin toxicities were assessed using National Cancer Institute Common Toxicity Criteria (version 3) and compared using a 2-sample Wilcoxon matched-pairs rank test adjusted for tied values. RESULTS: Fifty consecutive patients were included between April 2005 and January 2007. Nail toxicity was significantly lower in the FS-protected foot compared with the control foot (grade 0: 100% versus 79%; and grade 1 and 2: 0% versus 21%, respectively) (P= .002). Skin toxicity was grade 0: 98% versus 94%; and grade 1 and 2: 2% versus 6% in the FS-protected and the control feet, respectively. The median times until toxicity occurrence were not found to differ significantly between the groups. One patient experienced discomfort because of cold intolerance. CONCLUSIONS: Cold therapy using FS significantly reduced the incidence of docetaxel-induced foot nail toxicity, as previously demonstrated using frozen gloves for the hands. PMID- 18286528 TI - Outcome of patients who develop acute leukemia or myelodysplasia as a second malignancy after solid tumors treated surgically or with strategies that include chemotherapy and/or radiation. AB - BACKGROUND: Evaluation of therapeutic outcomes and risk factors was undertaken for patients with primary solid tumors (PST) developing acute leukemia or myelodysplasia (MDS) as a second malignancy. METHODS: In all, 131 consecutive patients presenting to a single institution with leukemia or MDS after treatment for PST with surgery or chemotherapy/radiotherapy were examined. Management of the secondary acute leukemia and MDS consisted either of intensive therapy including allogeneic blood and marrow transplants or supportive measures. RESULTS: The time from diagnosis of PST to development of acute leukemia or MDS, the cytogenetic profile of patients, and their survival were similar irrespective of PST therapy with surgery alone or strategies involving chemotherapy and/or radiation. The median survival of all 131 patients was 10.5 months with a 5-year survival of 15.6%. Induction therapy and/or transplantation resulted in a median survival of 13.6 months and a 5-year survival of 26.6% compared with 6.5 months and 2% with supportive measures. Subset analysis of transplant recipients revealed a median survival of 17.6 months and a 37.9% 5-year survival. Despite a significantly lower recurrence rate the survival of transplant recipients was not improved secondary to a higher treatment-related mortality (TRM) rate. CONCLUSIONS: Patients developing acute leukemia or MDS after PST demonstrated similar cytogenetic profiles and clinical outcomes independent of the type of treatment. Survival was significantly better for patients able to undergo intensive therapy compared with supportive measures. The low recurrence rate for allograft recipients was consistent with a potent antileukemic effect that may translate into a survival benefit if TRM could be reduced. PMID- 18286530 TI - A nomogram predicting long-term biochemical recurrence after radical prostatectomy. AB - BACKGROUND: Men who undergo radical prostatectomy (RP) are at long-term risk of biochemical recurrence (BCR). In this report, the authors have described a model capable of predicting BCR up to at least 15 years after RP that can adjust predictions according to the disease-free interval. METHODS: Cox regression was used to model the probability of BCR (a prostate-specific antigen level>0.1 ng/mL and rising) in 601 men who underwent RP with a median follow-up of 11.4 years. The statistical significance of nomogram predictors was confirmed with a competing-risks regression model. The model was validated internally with 200 bootstraps and externally at 5 years, 10 years, and 15 years in 2 independent cohorts of 2963 and 3178 contemporary RP patients from 2 institutions. RESULTS: The 5-year, 10-year, 15-year, and 20-year actuarial rates of BCR-free survival were 84.8%, 71.2%, 61.1%, and 58.6%, respectively. Pathologic stage, surgical margin status, pathologic Gleason sum, type of RP, and adjuvant radiotherapy represented independent predictors of BCR in both Cox and competing-risks regression models and constituted the nomogram predictor variables. In internal validation, the nomogram accuracy was 79.3%, 77.2%, 79.7%, and 80.6% at 5 years, 10 years, 15 years, and 20 years, respectively, after RP. In external validation, the nomogram was 77.4% accurate at 5 years in the first cohort and 77.9%, 79.4%, and 86.3% accurate at 5 years, 10 years, and 15 years, respectively, in the second cohort. CONCLUSIONS: Patients who undergo RP remain at risk of BCR beyond 10 years after RP. The nomogram described in this report distinguishes itself from other tools by its ability to accurately predict the conditional probability of BCR up to at least 15 years after surgery. PMID- 18286529 TI - Imprinted tumor suppressor genes ARHI and PEG3 are the most frequently down regulated in human ovarian cancers by loss of heterozygosity and promoter methylation. AB - BACKGROUND: Imprinted tumor suppressor genes may be particularly important in the pathogenesis of ovarian cancer. Two imprinted genes, paternally expressed 3 (PEG3) and aplasia Ras homologue member I (ARHI), are the most frequently down regulated in ovarian cancers on gene expression arrays. METHODS: PEG3 and ARHI expression levels were evaluated with real-time reverse-transcriptase polymerase chain reaction (PCR) analysis. Promoter methylation was measured by pyrosequencing, and loss of heterozygosity (LOH) was detected by PCR-LOH assays. RESULTS: PEG3 was down-regulated in 75% and ARHI was down-regulated in 88% of 40 ovarian cancers. ARHI CpG islands I and II were hypermethylated in 13 of 42 ovarian cancers (31%) and in 5 of 42 ovarian cancers (12%), respectively, and hypermethylation was associated with reduced ARHI expression in all 18 samples of ovarian cancer with CpG island hypermethylation. PEG3 was hypermethylated in 11 of 42 ovarian cancers (26%), and PEG3 expression was down-regulated in 10 of those 11 cancers. LOH was detected in 8 of 35 informative cases for ARHI (23%) and in 5 of 25 informative cases for PEG3 (20%). PEG3 and ARHI expression was highly correlated in human ovarian cancers (correlation coefficient [R]=0.69; P< .0001). PEG3 and ARHI also were methylated concordantly in ovarian cancers (R=0.36; P= .019). Re-expression of PEG3, similar to that of ARHI, markedly inhibited ovarian cancer growth. ARHI and PEG3 expression could be restored by treatment with 5-aza-2'-deoxycytidine and trichostatin A, consistent with the importance of promoter methylation and histone acetylation in regulating expression of both genes. CONCLUSIONS: Loss of expression of the growth inhibitory imprinted genes ARHI and PEG3 through promoter methylation, LOH, and other mechanisms may stimulate clonogenic growth and contribute to the pathogenesis of a majority of ovarian cancers. PMID- 18286531 TI - Multiple gene methylation of nonsmall cell lung cancers evaluated with 3 dimensional microarray. AB - BACKGROUND: Aberrant DNA methylation of the CpG islands for cancer-related genes is among the earliest and most frequent alterations in cancer and may be useful for diagnosing cancer or evaluating recurrent disease. METHODS: In this study, a 3-dimensional (3-D), polyacrylamide gel-based DNA microarray coupled with linker polymerase chain reaction (PCR) was developed to detect hypermethylation of CpG islands in multiple genes from a large group of different samples. The authors determined the frequency of aberrant promoter methylation of 15 genes in 28 resected primary nonsmall cell lung cancers (NSCLCs) and in 12 corresponding nonmalignant lung tissues. RESULTS: Methylation frequencies in the tumor samples were detected in 18% of samples for the breast cancer 1 gene BRCA1, in 43% of samples for the tissue inhibitor of metalloproteinase 3 gene TIMP-3, in 38% of samples for the cyclin-dependent kinase inhibitor 4A gene p16INK4a, in 54% of samples for the cadherin 13 gene CDH13, in 50% of samples for the death associated protein kinase gene DAPK, in 11% of samples for the E-cadherin gene ECAD, in 25% of samples for the insulin-like growth factor binding protein 7 gene IGFBP7, in 18% of samples for the Ras association domain family 1 gene RASSF1, in 68% of samples for the adenomatous polyposis coli gene APC, in 7% of samples for the cyclin-dependent kinase inhibitor gene p15, in 18% of samples for the CD44 cell adhesion molecule gene, in 29% of samples for the human Mut-L homolog gene hMLH, in 32% of samples for the human telomerase reverse transcriptase gene hTERT, in 64% of samples for the calcitonin gene-related polypeptide alpha gene CALCA, and in 54% of samples for the estrogen receptor gene ER; however, methylation was not observed in the majority of corresponding nonmalignant tissues. Six samples in from 28 tumors had >6 genes methylated, and 1 sample had 13 genes methylated. Methylation of these genes was correlated with some clinicopathologic patient characteristics. CONCLUSIONS: This study demonstrated that a 3-D microarray could be used to detect DNA hypermethylation and provided a high-throughput platform for DNA hypermethylation analysis. PMID- 18286532 TI - Dying with cancer: the influence of age, comorbidity, and cancer site. AB - BACKGROUND: Cancer survival is influenced by age, comorbidity, and type of cancer. A population-based study was conducted to compare the interplay between age and mortality for different cancers. METHODS: This study analyzed 784,378 cases, comprising 22 of the commonest SEER cancers diagnosed between 1984 and 1993. Competing hazards and proportional hazard analyses for cancer-specific and comorbid death were performed. RESULTS: Median follow-up was up to 159 months, and the median age of diagnosis was 67 years. Cancer-specific and comorbid deaths accumulated most within the first years of diagnosis. With the more biologically aggressive cancers, cancer deaths invariably exceeded comorbid deaths. For the remaining 70% of cancers, comorbidity remained the dominant mode of death. Deaths attributable to both cancer and comorbidity accumulated mostly after the seventh decade of life. Cancer site had a 3-fold greater effect on overall survival than age at diagnosis and a 30-fold effect with cancer-specific survival; age at diagnosis had a 5-fold greater effect on comorbid deaths than site. CONCLUSIONS: Both the age of the affected individual and the biology of the particular cancer have major influences on cancer survival and mode of death. Cancer is largely a disease of the elderly. Within affected individuals, fatalities attributable to cancer and comorbidity appeared inter-related, with cancer-specific deaths dominating for more lethal cancers and comorbid deaths dominating for the remaining majority. For these reasons, further improvements in overall survival may be best anticipated from better geriatric and general medical management as much as from better cancer management. PMID- 18286534 TI - Distinction between isolated tumor cells and micrometastases in breast cancer: is it reliable and useful? AB - BACKGROUND: In routine practice, the distinction between isolated tumor cells (ITC) and micrometastases (MIC) in patients with breast cancer is sometimes difficult to discern. The authors assessed differences in classifying patients according to the American Joint Commission on Cancer (AJCC) and the International Union Against Cancer (UICC) definitions and method of sizing. METHODS: We assessed the characteristics of metastatic deposits in only 1 involved lymph node in 337 patients with operable breast cancer (median follow-up, 15.3 years). When sizing multiple clusters, either the diameter of the area with close clusters (Method 1) or the size of the largest cluster (Method 2) was taken into account. Patients were classified and their survival was assessed according to the 2 sizing methods and the criteria used for definitions (size in AJCC; size and topography in UICC). RESULTS: With the AJCC definitions, 32 patients would be differently classified according to the method of sizing. With the UICC definitions, some patients with parenchymal ITC would be classified as pN1mi, 38 by Method 1 and 53 by Method 2. Some pathologists would classify the 66 patients who had isolated capsular vascular invasion as pN0. Classification was uncertain in 136 (40%) to 151 (45 %) patients. Survival was not significantly different between pN0(i+) and pN1(mi) patients. CONCLUSIONS: The distinction between ITC and MIC was often difficult and without any prognostic significance. Precise guidelines are more useful for staging than for therapy. Thus, complete axillary dissection is usually performed in pN0(i+) and pN1(mi) patients, whereas chemotherapy is not indicated or debatable when MIC is the only 1 pejorative criterion. PMID- 18286535 TI - Accuracy of fine-needle aspiration cytology of axillary lymph nodes in breast cancer patients: a study of 115 cases with cytologic-histologic correlation. AB - BACKGROUND: Fine-needle aspiration (FNA) cytology of axillary lymph nodes is a simple, minimally invasive technique that can be used to improve preoperative determination of the status of the axillary lymph nodes in patients with breast cancer, thereby serving as a tool with which to triage patients for sentinel versus full lymph node dissection procedures. The aim of the current study was to determine the sensitivity and specificity of FNA cytology to detect metastatic breast carcinoma in axillary lymph nodes. METHODS: A total of 115 FNAs of axillary lymph nodes of breast cancer patients with histologic follow-up (subsequent sentinel or full lymph node dissection) were included in the current study. The specificity and sensitivity, as well as the positive and negative predictive values, were calculated. RESULTS: The positive and negative predictive values of FNA cytology of axillary lymph nodes for metastatic breast carcinoma were 1.00 and 0.60, respectively. The overall sensitivity of axillary lymph node FNA in all the cases studied was 65% and the specificity was 100%. The sensitivity of FNA was lower in the sentinel lymph node group than in the full lymph node dissection group (16% vs 88%, respectively), which was believed to be attributable to the small size of the metastatic foci in the sentinel lymph node group (median, 0.25 cm). All false-negative FNAs, with the exception of 1 case, were believed to be the result of sampling error. There was no 'true' false positive FNA case in the current study. CONCLUSIONS: FNA of axillary lymph nodes is a sensitive and very specific method with which to detect metastasis in breast cancer patients. Because of its excellent positive predictive value, full axillary lymph node dissection can be planned safely instead of a sentinel lymph node dissection when a preoperative positive FNA result is rendered. . PMID- 18286536 TI - Gemcitabine and vinorelbine in pemetrexed-pretreated patients with malignant pleural mesothelioma. AB - BACKGROUND: Pemetrexed-cisplatin chemotherapy is the standard of care in the first-line treatment of unresectable malignant pleural mesothelioma (MPM). Second line cytotoxic therapy is considered for a growing group of patients, but the optimal treatment has not been defined to date. Gemcitabine and vinorelbine have shown activity in the first-line setting. The objective of this study was to evaluate the activity and toxicity of the gemcitabine-vinorelbine combination in pemetrexed-pretreated patients with MPM. METHODS: From January 2004 to September 2006, 30 consecutive patients who were pretreated with pemetrexed with or without a platinum-derivative were enrolled. Gemcitabine 1000 mg/m(2) and vinorelbine 25 mg/m(2) were administered intravenously on Days 1 and 8 every 3 weeks. Treatment was repeated for a maximum of 6 cycles or until progression or unacceptable toxicity. RESULTS: A partial response was observed in 3 patients (10%; 95% confidence interval [CI], 2.1-26.5%), and 10 patients (33.3%; 95% CI, 17.3-52.8%) had stable disease after treatment. Overall, 13 patients (43.3%; 95% CI, 25.5 62.6%) achieved disease control. The median time to progression was 2.8 months (range, 0.6-12.1 months), and the median survival was 10.9 months (range, 0.8 25.3 months). Hematologic toxicity was acceptable, with grade 3 or 4 neutropenia occurring in 11% of patients and thrombocytopenia occurring in 4% of patients; no case of febrile neutropenia was observed. Nonhematologic toxicity generally was mild. CONCLUSIONS: The gemcitabine and vinorelbine combination was moderately active and had an acceptable toxicity profile in pemetrexed-pretreated patients with MPM. The role of second-line treatment in MPM needs to be evaluated in prospective trials in large series of patients who are stratified according to previous treatment and prognostic factors. PMID- 18286537 TI - Isolation and characterization of a putative liver progenitor population after treatment of fetal rat hepatocytes with TGF-beta. AB - The "in vitro" establishment of a physiological model of bipotential liver progenitors would be useful for analyzing the molecular mechanisms involved in regulating growth and differentiation, as well as studying their potential role/s in liver physiology and pathology. The transforming growth factor-beta (TGF-beta) induces de-differentiation of fetal rat hepatocytes (FH), concomitant with changes in morphology. The aim of this work was to isolate and characterize this population of TGF-beta-treated fetal hepatocytes (TbetaT-FH) and test whether they can behave as liver progenitors. The TbetaT-FH isolated cell lines show high expression of Thy-1 and low expression of c-Kit. They express liver-specific proteins, such as albumin and alpha-fetoprotein, and mesenchymal markers, such as vimentin. TbetaT-FH maintain expression of the hnf3beta gene, but lose expression of hnf1beta, hnf4, and hnf6. They express c-met and show an increase in proliferation in response to HGF. Interestingly, the transdifferentiation process is coincident with changes in the expression of genes related to the oxidative metabolism. TbetaT-FH cultured in the presence of EGF + DMSO change morphology, towards epithelial cells, gaining expression of CK19 and c-Kit, markers found in hepatoblasts and bile duct cells. Furthermore, TbetaT-FH form duct-like structures when cultured on Matrigel. TbetaT-FH show also potential to revert to an hepatocyte phenotype when submitted to a long-term "in vitro" differentiation protocol towards hepatocytic lineage. In summary, our results support the hypothesis that hepatocytes can function as facultative liver stem cells and demonstrate that TGF-beta might play an essential role in the transdifferentiation process. PMID- 18286539 TI - Bag3 gene expression is regulated by heat shock factor 1. AB - BAG3 protein, a member of the BAG co-chaperones family, sustains cell survival, through its interaction with the heat shock protein (HSP) 70, in a variety of normal and neoplastic cell types. bag3 gene expression is induced by stressful stimuli. Here we report for the first time that two of the three putative heat shock-responsive elements (HSEs) in bag3 promoter interact with the heat shock factor (HSF) 1 in vitro and in vivo. Furthermore, downmodulation of HSF1 protein levels by specific small interfering (si) RNAs results in reducing BAG3 protein levels, indicating that the transcription factor plays a major role in bag3 gene expression. Because of the anti-apoptotic role of BAG3 protein, these results disclose a previously unrecognized pathway, through which HSF1 maintains cell survival. PMID- 18286538 TI - Indian hedgehog promotes the migration of rat activated pancreatic stellate cells by increasing membrane type-1 matrix metalloproteinase on the plasma membrane. AB - Indian hedgehog (Ihh) is a member of hedgehog peptides family that exerts diverse effects on multiple cellular functions. Since Ihh expression is elevated in the pancreas of chronic pancreatitis patients, Ihh has been assumed to participate in the chronic pancreatic injury, especially in pancreatic fibrosis. However, its function in pancreatic fibrosis is still unknown. We thus examined Ihh effects on rat activated pancreatic stellate cells (PSCs) that play a central role in pancreatic fibrosis. Activated PSCs express both patched-1 and smoothened that are essential components of hedgehog receptor system. Ihh did not alter the PSC expression of collagen-1 or alpha-smooth muscle actin, a parameter of PSC transformation, or did not change PSC proliferation. However, Ihh enhanced PSC migration in both chemotactic and chemokinetic manners. Furthermore, Ihh increased the amount of membrane-type 1 matrix metalloproteinase (MT1-MMP) and altered its localization on the plasma membrane, which plays a stimulatory role in cellular migration. In addition, tissue inhibitor of metalloproteinase-2 (TIMP 2) attenuated Ihh-stimulated PSC migration. Since most hedgehog intracellular signals are mediated by Gli-1 transcription factor, we investigated its contribution to Ihh-enhancement of PSC migration. Ihh induced Gli-1 nuclear accumulation in PSCs, indicating that Ihh stimulates Gli-1-dependent signaling pathway in PSCs. Unexpectedly, however, adenovirus-mediated Gli-1 overexpression blocked the Ihh enhancement of both MT1-MMP localization on the plasma membrane and PSC migration. Furthermore, reduction of Gli-1 expression with RNA interference augmented Ihh-stimulated PSC migration. These data indicate that Ihh promotes PSC migration by enhancing MT1-MMP localization on the plasma membrane but is negatively regulated by Gli-1. PMID- 18286540 TI - FGFR3 contributes to intestinal crypt cell growth arrest. AB - Fibroblast growth factors (FGFs) are important regulators of the dynamic development and turnover of tissues. Among FGF receptors, FGFR3 expression is confined in the intestinal crypts. We examined FGFR3-deficient mice and saw increased intestinal crypt depth but no change in villae length or in the distribution of differentiated intestinal cells, suggesting that the impact of lack of FGFR3 was limited to the progenitor cell compartment. Accordingly, enhancement of intestinal crypt proliferation was observed in FGFR3 mutant mice and interestingly, upon anti-FGFR3 antibody administration in wild type mice. Moreover, injection of FGF18, a ligand of FGFR3, in wild type mice resulted in decreased cell proliferation within the intestinal crypts. In addition, we found that ERK level of activation was increased in FGFR3-deficient intestinal epithelium. In vitro studies showed that ERK, AKT and activation was regulated by FGFs and that ERK level of activation was inversely correlated to FGFR3 level of expression in the intestinal crypt cells. Furthermore, effects of FGF18 on ERK and AKT activation paralleled FGFR3 effects on these intracellular targets. Our data indicate that FGF18 and FGFR3 are involved, possibly as partners, in the control of intestinal precursor cell proliferation. PMID- 18286541 TI - CYR61 is a novel gene associated with temperature-dependent changes in fish metabolism as revealed by cDNA microarray analysis on a medaka Oryzias latipes cell line. AB - A microarray comprising 3,514 cDNAs was constructed from a medaka EST library to elucidate the transcriptional responses associated with temperature shift from 25 to 15 degrees C in a medaka cell line. Microarray analysis revealed that the mRNA levels of 313 clones were significantly different in at least one combination of different incubation periods up to 7 days at a given incubation temperature or between 25 and 15 degrees C at a given incubation period (P < 0.05). These genes are known to be associated with various biological processes including morphogenesis, cell proliferation and response to stress. A number of genes encoding proteins which localize in extracellular areas were apparently up regulated at 15 degrees C, whereas those localizing in intracellular areas were down-regulated at this temperature. In addition, while a number of genes represented long-term expression changes, only a few responded to short-term inductions. A typical example was CYR61, a multifunctional matricellular signaling modulator, the mRNA levels of which increased after temperature shift from 25 to 15 degrees C in 3 h, and then decreased rapidly to near the original level within 12 h. Another series of analyses by quantitative reverse transcription-PCR revealed that the mRNA levels of CYR61 at 5 degrees C were significantly higher even at 24 h after temperature shift compared to those of the cells successively maintained at 25 degrees C. These analyses suggest that remodeling and reorganizing of extracellular structure of cells are important to offset the low temperature effect and CYR61 is considered to be a novel gene associated with temperature response in poikilotherms. PMID- 18286543 TI - Coexpression of osteogenic and adipogenic differentiation markers in selected subpopulations of primary human mesenchymal progenitor cells. AB - Knowledge of the basic mechanisms controlling osteogenesis and adipogenesis might provide new insights into the prevention of osteoporosis and age-related osteopenia. With the help of magnetic cell sorting and fluorescence activated cell sorting (FACS), osteoblastic subpopulations of mesenchymal progenitor cells were characterized. Alkaline phosphatase (AP) negative cells expressed low levels of osteoblastic and adipocytic markers. AP positive cells expressed adipocytic markers more strongly than the AP negative cell populations, thus suggesting that committed osteoblasts exhibit a greater adipogenic potential. AP negative cells differentiated to the mature osteoblastic phenotype, as demonstrated by increased AP-activity and osteocalcin secretion under standard osteogenic culture conditions. Surprisingly, this was accompanied by increased expression of adipocytic gene markers such as peroxisome proliferator-activated receptor gamma2, lipoprotein lipase and fatty acid binding protein. The induction of adipogenic markers was suppressed by transforming growth factor-beta1 (TGF-beta1) and promoted by bone morphogenetic protein 2 (BMP-2). Osteogenic culture conditions including BMP-2 induced both the formation of mineralized nodules and cytoplasmic lipid vacuoles. Upon immunogold electron microscopic analysis, osteoblastic and adipogenic marker proteins were detectable in the same cell. Our results suggest that osteogenic and adipogenic differentiation in human mesenchymal progenitor cells might not be exclusively reciprocal, but rather, a parallel event until late during osteoblast development. PMID- 18286542 TI - The effects of cocaine on regional brain glucose metabolism is attenuated in dopamine transporter knockout mice. AB - Cocaine's ability to block the dopamine transporter (DAT) is crucial for its reinforcing effects. However the brain functional consequences of DAT blockade by cocaine are less clear since they are confounded by its concomitant blockade of norepinephrineand serotonin transporters. To separate the dopaminergic from the non-dopaminergic effects of cocaine on brain function we compared the regional brain metabolic responses to cocaine between dopamine transporter deficient (DAT( /-)) mice with that of their DAT(+/+) littermates. We measured regional brain metabolism (marker of brain function) with 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) and microPET (muPET) before and after acute cocaine administration (i.p. 10 mg/kg). Scans were conducted 2 weeks apart. At baseline DAT(-/-) mice had significantly greater metabolism in thalamus and cerebellum than DAT(+/+). Acute cocaine decreased whole brain metabolismand this effect was greater in DAT(+/+) (15%) than in DAT(-/-) mice (5%). DAT(+/+) mice showed regional decreases in the olfactory bulb, motor cortex, striatum, hippocampus, thalamus and cerebellum whereas DAT(-/-) mice showed decreases only in thalamus. The differential pattern of regional responses to cocaine in DAT(-/-) and DAT(+/+) suggests that most of the brain metabolic changes from acute cocaine are due to DAT blockade. Cocaine induced decreases in metabolism in thalamus (region with dense noradrenergic innervation) in DAT(-/-) suggest that these were mediated by cocaine's blockade of norepinephrine transporters. The greater baseline metabolism in DAT(-/-) than DAT(+/+) mice in cerebellum (brain region mostly devoid of DAT) suggests that dopamine indirectly regulates activity of these brain regions. PMID- 18286544 TI - Photoelectron circular dichroism: Chiral asymmetry in the angular distribution of electrons emitted by (+)-S-carvone. AB - The technique of photoelectron circular dichroism (PECD) is introduced and illustrated by the presentation of results obtained for the C 1s core ionization of (+)-S-carvone enantiomers. Using circularly polarized ionizing radiation, large chiral effects in the angular distribution of photoelectrons emitted from a dilute, randomly oriented gas phase sample can be detected. This effect is predicted in the pure electric dipole approximation and is expected to be quite general. The forward-backward asymmetry regularly exceeds 10%, as demonstrated here for the carbonyl C 1s photoemission of carvone. Calculations are presented that reveal a pronounced dependence of this effect on molecular conformation. In the case of carvone the PECD associated with the C 1s electron localized at the carbonyl group varies with orientation of the isopropenyl tail group at the opposite end of the molecule. Comparisons of calculations with experiment confirm the molecular configuration and permit inferences to be drawn concerning the relative conformer populations in the experimental sample. PMID- 18286545 TI - Pseudoasymmetry: a final twist? AB - The original definition of "pseudoasymmetry" conveyed the apparent paradox that a tetrahedral center with four different groups did not result in overall chirality. However, there are problems in applying the concept to cyclic systems that do not contain chirotopic centers. Pseudoasymmetry appears most appropriate to acyclic systems with chirotopic carbon centers, e.g. the meso trihydroxyglutaric acids. Analogous cyclic cases, e.g. the isomeric 1,4 dimethylcyclobutanes, are best treated as diastereomers, and may indeed be described by an interesting extension of the like-unlike notation. Remarkably, in several tri- and tetramethylcyclohexanes, CIP descriptors cannot be applied even to chirotopic centers, which can only be described by the modified l-u notation. PMID- 18286546 TI - Severe hypercholesterolemia mediated by lipoprotein X in a pediatric patient with chronic graft-versus-host disease of the liver. AB - We describe a case of extreme hypercholesterolemia, mediated by lipoprotein X, in a 12-year-old Caucasian female who underwent an unrelated allogenic bone marrow transplant for relapsed acute myelocytic leukemia (AML). Her post-transplant course was complicated by severe chronic graft-versus-host disease (GVHD) of the liver. Previously normal serum cholesterol and triglycerides rose to 1,122 mg/dl (29.0 mmol/L) and 1,100 mg/dl (12.4 mmol/L), respectively. Serum cholesterol appeared to be dominantly carried by lipoprotein X. Intra-hepatic cholestasis leading to reflux of bile lipoproteins into the blood stream and subsequent formation of lipoprotein X appears to be the mechanism underlying this phenomenon. PMID- 18286547 TI - Activation of the ERK pathway in osteoblastic cells, role of gremlin and BMP-2. AB - Gremlin is a glycoprotein that binds and antagonizes the actions of bone morphogenetic proteins (BMPs) -2, -4, and -7. Gremlin appears to activate the extracellular regulated kinase (ERK) pathway in endothelial and tumor cells, and as a consequence to have direct cellular effects. To determine whether gremlin has direct effects in osteoblasts, independent of its BMP binding activity, we examined its effects in ST-2 murine stromal cell lines and in primary cultures of murine calvarial osteoblasts. Gremlin did not activate Signaling mothers against decapentaplegic (Smad), and suppressed the BMP-2 induced Smad 1/5/8 phosphorylation and the transactivation of the BMP/Smad reporter construct 12xSBE Oc-pGL3, confirming its BMPs antagonizing activity. Neither gremlin nor BMP-2 induced ERK 1/2 activation in ST-2 cells or calvarial osteoblasts. Moreover, slight changes in culture conditions induced the phosphorylation of ERK independent from BMP or gremlin exposure. In conclusion, gremlin inhibits BMP-2 signaling and activity, and does not have independent actions on ERK signaling in osteoblasts. Consequently, gremlin activity in osteoblasts can be attributed only to its BMP antagonizing effects. PMID- 18286548 TI - A longitudinal assessment of the effect of inhaled fluticasone propionate therapy on adrenal function and growth in young children with asthma. AB - OBJECTIVE: Fluticasone proprionate (FP) is increasingly used to treat very young children with asthma. Its safety in terms of effects on the hypothalamic pituitary axis (HPA) and growth in this age group is uncertain. PATIENTS AND METHODS: Eleven children (median (range) age 10 (5.6-24.3) months) presenting with recurrent wheeze and family history of asthma were studied prospectively for a period of 18 months. Children received daily-inhaled FP 250 microg via a spacer device. No other corticosteroid therapy was administered prior to or during the study. A Short Standard Synacthen Test (SST) (125 microg) was performed pretreatment, and after 6 and 18 months. Weight (Wt), height (Ht), and body mass index (BMI) were measured at 3-6 monthly intervals. RESULTS: Fasting early morning and peak cortisol levels remained within the normal reference range with therapy. There were no changes in Ht SDS, whereas both Wt SDS (baseline 0.05 ( 2.17 to 0.52) vs. +18 months 0.68 (-0.5 to 1.36) P < 0.02) and BMI SDS (-0.22 ( 1.73 to 0.75) vs. 0.86 (0.03 to 1.99) P < 0.005) increased after 18 months of treatment. CONCLUSION: Daily treatment with inhaled FP 250 microg in young children with asthma appears to have no adverse effects on the HPA or on linear growth, however, treatment is associated with increases in body Wt and BMI in young children. PMID- 18286549 TI - Use of balloon-expandable metallic stent in a premature infant with congenital tracheobronchial stenosis. AB - A congenital tracheobronchial stenosis is a rare obstructive lesion of the airway characterized by intrinsic narrowing of a segment of the trachea and bronchi. In this report, we present the smallest premature infant with a congenital tracheobronchial stenosis who has been successfully treated with balloon dilatation and placement of a balloon-expandable metallic stent. PMID- 18286550 TI - Airway stents in children. AB - Airway obstruction in children is a rare, but difficult clinical problem, with no clear agreement on optimal therapeutic approach. Stenting of the airway has been used successfully in adults, and is an attractive alternative in children. Fundamental differences of pediatric compared to adult use include the benign nature of most stenoses, the narrow and soft airways of children, the required long-term tolerance and adaptation to growth. These differences may significantly alter the therapeutic balance, calling into question the precise role stents play in the treatment of airway obstruction in children. Stent placement can be technically demanding but is not exceedingly difficult. Experience is necessary to select the proper size and type of stent. Metal stents usually achieve airway patency and clinical improvement in the majority of cases, while this is less frequently the case with silicone stents. Some complications such as granulation and secretion retention seem to occur in most children after stent implantation. Unfortunately, severe complications including death have been reported in a significant proportion of children. Stent related mortality can be estimated at 12.9% from published data, but these include complication centered reports. The initial euphoria for airway stents in children has largely abated and most authors agree that they should only be employed in circumstances with no good alternatives. It is crucial that all surgical and medical alternatives are considered and the decision to place a stent is not made because other options are overlooked or not available locally. Stent use in a palliative setting has also been reported and is probably reasonable. Stents will only allow limited adaptation for the growth of pediatric airways by balloon dilatation. All metal stents should be considered as potentially permanent, and removal sometimes may only be possible through a surgical and sometimes risky approach. PMID- 18286551 TI - IL-10 gene polymorphism at -1082 A/G is associated with severe rhinovirus bronchiolitis in infants. AB - We analyzed polymorphisms of IL-10 -1082 G/A, IL-18 -137 G/C, TLR4 +896 A/G, and IFNG +874 T/A in 139 infants under 6 months of age hospitalized with bronchiolitis and 400 unselected blood donors. Causative viruses were determined by PCR. Infants with bronchiolitis associated with a virus other than respiratory syncytial virus (N = 18), were more often IL-10 -1082 allele G non-carriers, that is, homozygous for allele A (AA) than controls (66.7% vs. 28.0%, P < 0.0001). Infants with RSV bronchiolitis did not differ from controls. This finding suggests a different pathogenic mechanism for RSV bronchiolitis as compared with wheezing associated with other viral infections, for example, rhinovirus in infants under 6 months of age. PMID- 18286552 TI - Photodissociation of hydrated hydrogen iodide clusters. AB - Using high-level ab initio calculations and excited state ab initio molecular dynamics simulations, we show that hydrated iodic acids release hydrogen radicals and/or hydrogen molecules as well as iodine radicals upon excitation. Its photoreaction process involving charge transfer to the solvent takes place in four steps: 1) hydration of the acid, 2) charge transfer to water upon excitation of hydrated acid, 3) detachment of the neutral iodine atom, and 4) detachment of the hydrogen radical. The iodine detachment process from excited hydrated hydro iodic acids is exothermic and the detachment of hydrogen radicals from hydrated hydronium radicals is spontaneous if the initial kinetic energy of the cluster is high enough to get over the activation barrier of the detachment. The complete release of the radicals can be understood in terms of kinetics. This study shows how the hydrogen and halogen radicals are dissociated and released from their hydrated acids. Simple experiments corroborate our predicted mechanism for the release of hydrogen molecules from iodic acid in water by ultraviolet light. PMID- 18286553 TI - Targeted RNA interference of cyclin A2 mediated by functionalized single-walled carbon nanotubes induces proliferation arrest and apoptosis in chronic myelogenous leukemia K562 cells. AB - Cyclin A(2) plays critical role in DNA replication, transcription, and cell cycle regulation. Its overexpression has been detected and related to many types of cancers including leukemia, suggesting that suppression of cyclin A(2) would be an attractive strategy to prevent tumor progression. Herein, we apply functionalized single wall carbon nanotubes (f-SWNTs) to carry small interfering RNA (siRNA) into K562 cells and determine whether inhibition of cyclin A(2) would be a potential therapeutic target for chronic myelogenous leukemia. The results show functionalized SWNTs can facilitate the coupling of siRNA specifically targeting human cyclin A(2) to form cyclin A(2) siRNA-f-SWNTs complexes. These functionalized SWNTs readily enter K562 cells, resulting in suppression of cyclin A(2) expression. We demonstrate that depletion of cyclin A(2) in this manner inhibits cell proliferation and promotes apoptosis, and cyclin A(2) can serve as a novel therapeutic target. siRNA against cyclin A(2) delivered by functionalized single wall carbon nanotubes may be a useful therapeutic strategy for chronic myelogenous leukemia cells. This would provide new insights on additional therapeutic options for chronic myelogenous leukemia beyond chemotherapy in light of increasing multidrug resistance. PMID- 18286554 TI - Analytic models of domain-averaged Fermi holes: a new tool for the study of the nature of chemical bonds. AB - Simple analytical models are introduced that significantly enhance the ability to understand and rationalise the nature of bonding interactions depicted by domain averaged Fermi hole (DAFH) analysis. The examples presented show that besides shedding new light on the role of electron-sharing in ordinary two-centre two electron (2c-2e) chemical bonds that are well represented by the classical Lewis model, the proposed approach also provides interesting new insights into the nature of bonding interactions that go beyond the traditional Lewis paradigm. This is, for example, the case of 3c-2e multicentre bonding, but a straightforward extension of the approach also reveals for direct metal-metal bonding the existence of a completely new type of bonding interaction that involves the mutual exchange of electrons between the lone pairs on adjacent metal atoms. PMID- 18286555 TI - A novel green template for the synthesis of mesoporous silica. AB - Mesoporous pure silicas and functionalized silica with a narrow pore size distribution centered at 3.8 nm were prepared by a novel template, amphiphilic dendritic polyglycerol. The resulting silica materials were characterized by electron microscopy; nitrogen adsorption; (1)H, (13)C, and (29)Si solid-state cross-polarization magic-angle spinning NMR spectroscopy. It was shown that the template could be completely removed from the pure and functionalized silica in an environmentally friendly way by means of a simple water extraction procedure. Furthermore, it was shown that these materials could be easily functionalized, for example, by employing aminopropyl groups. Thus, a new environmentally friendly pathway to this fascinating class of silica material has been opened. PMID- 18286556 TI - Subnanometer-resolved patterning of bicomponent self-assembled monolayers on Au(111). PMID- 18286557 TI - Aggregation of ZnO nanocrystallites for high conversion efficiency in dye sensitized solar cells. PMID- 18286558 TI - Efficient oxidative alkyne homocoupling catalyzed by a monomeric dicopper substituted silicotungstate. PMID- 18286559 TI - Ru-catalyzed tandem cross-metathesis/intramolecular-hydroarylation sequence. PMID- 18286560 TI - An electroinitiated cation chain reaction: intramolecular carbon-carbon bond formation between thioacetal and olefin groups. PMID- 18286561 TI - Metastable dissociation of anions formed by electron attachment. AB - A comprehensive analysis of metastable dissociation of 2,4-dinitrotoluene (DNT) parent anions formed by attachment of electrons of controlled energy is presented. We characterize the energy dependence and kinetic energy release of the reaction which competes with autodetachment. A surprising finding is a highly exothermic metastable reaction triggered by the attachment of thermal electrons which we relate to the well-known electrostatic ignition hazards of DNT and other explosives. Quantum chemical calculations are performed for dinitrobenzene in order to elucidate the process of NO abstraction. PMID- 18286562 TI - Stability of alkali metal halide polymorphs as a function of pressure. AB - We investigated the regions of thermodynamic stability of possible modifications of the alkali metal halides as a function of pressure and type of alkali metal and halogen. Both Hartree-Fock and density functional calculations (for six different functionals) were performed. The results are in good agreement with experiment, and the trends in the computed quantities such as transition pressures and lattice parameters as a function of the ab initio method are similar to those found in earlier studies of the alkali metal sulfides. We predict that in most of these systems the so-called 5-5 modification should be metastable at standard pressure and be thermodynamically stable at slightly negative pressures. The sizes of the pressure ranges over which the various modifications are stable showed characteristic trends as a function of the type of the constituent elements, thus generalizing the traditional pressure-homologue rule for transition pressures and stable phases in ionic solids. PMID- 18286563 TI - Synthesis of syn-2,4-dimercapto-1,3,2,4-dithiadigermetane and its application to Ge2PdS4 cluster synthesis. AB - The sulfurization of DmpGeH(3) (Dmp=2,6-dimesitylphenyl) afforded the trinuclear germanium sulfide [DmpGe(mu-S)](2)(mu-S)(2)Ge(SH)-Dmp and a series of polythiadigermabicyclo[x.1.1]alkanes (x=3, 4, 5). The reduction of the S--S bonds of these germabicycloalkanes by NaBH(4) at 0 degrees C afforded the dinuclear mercaptogermane syn-[DmpGe(SH)(mu-S)(2)Ge(SH)-Dmp] (5) in good yield. The reaction of [Pd(dppe)Cl(2)] (dppe=1,2-bis(diphenylphosphanyl)ethane) and the dilithium salt of 5 prepared in situ by the addition of nBuLi (2 equiv) gave the Ge(2)PdS(4) cluster [DmpGe(mu-S)](2)[(mu-S)(2)Pd(dppe)], in which the dithiadigermetanedithiolate is bound to the Pd atom at the two thiolato sulfur atoms. The same reaction with [Pd(PPh(3))(2)Cl(2)] gave another Ge(2)PdS(4) cluster, [DmpGe(mu-S)](2)[(mu-S)(2)Pd(PPh(3))], but with the dithiadigermetanedithiolate and the Pd center conjoined through a mu-S atom between the two germanium atoms in addition to the two thiolato sulfur atoms to form a highly distorted cluster core. The formation of two different types of Ge(2)PdS(4) clusters represents the usefulness of 5 in the synthesis of various polynuclear complexes composed of germanium and transition metals. PMID- 18286564 TI - Hydrogen-bond-mediated asymmetric catalysis. AB - The utilization of hydrogen bonding as an activation force has become a powerful tool in asymmetric organocatalysis. Significant advances have been made in the recent past in this emerging field. Due to space constraints, this Focus Review summarizes only the key aspects with an emphasis on catalysis based on chiral ureas and thioureas, diols, and phosphoric acids. The examples provided neatly demonstrate that chiral ureas and thioureas, diols, and phosphoric acids display effective and unique activation modes of catalysis for a broad spectrum of asymmetric organic transformations, including single-step and multiple-step cascade reactions. These functionalities, which have the ability to afford efficient H-bond activation of electrophiles including C=O, C=N, aziridines, and epoxides, have established their status as "privileged" functional groups in the design of organocatalysts. PMID- 18286566 TI - A role for ARF6 in dendritic cell podosome formation and migration. AB - ADP-ribosylation factor 6 (ARF6) is a widely expressed GTPase that influences both membrane traffic and actin cytoskeleton function. Its role in dendritic cells (DC) has not previously been investigated. We analysed the effect of retroviral expression of ARF6 GDP/GTP binding and other functional mutants in primary murine DC. Maturation in response to lipopolysaccharide (LPS) proceeded normally in DC expressing ARF6 mutants and production of inflammatory cytokines was similarly unaffected. Although LPS-stimulated macropinocytosis was suppressed by expression of the GTP-binding Q67L ARF6 mutant we detected no overall activation of ARF6 by LPS. The ability of immature DC to migrate towards CCL3 and to a lesser extent, of mature DC to migrate towards CCL19, was compromised by expression of either the Q67L or the GDP-binding T44N mutant. Examination of the actin cytoskeleton in these cells revealed that both mutants strongly inhibited the formation of F-actin-rich podosomes, providing a possible explanation for the effects of ARF6 mutants on DC migration. Thus, these studies identify responses in DC that require normal ARF6 function, though not necessarily further ARF6 activation. They reveal for the first time a role for ARF6 in podosome formation and demonstrate functional effects of the T44N ARF6 mutant. PMID- 18286565 TI - Intradermal NKT cell activation during DNA priming in heterologous prime-boost vaccination enhances T cell responses and protection against Leishmania. AB - Heterologous prime-boost vaccination employing DNA-vaccinia virus (VACV) modality using the Leishmania homologue of receptors for activated C kinase (LACK) (p36) antigen has been shown to elicit protective immunity against both murine cutaneous and visceral leishmaniasis. However, DNA priming is known to have limited efficacy; therefore in the current study the effect of NKT cell activation using alpha-galactosyl-ceramide (alphaGalCer) during intradermal DNAp36 priming was examined. Vaccinated mice receiving alphaGalCer + DNAp36 followed by a boost with VVp36 appeared to be resolving their lesions and had at ten- to 20-fold higher reductions in parasite burdens. NKT cell activation during alphaGalCer + DNAp36 priming resulted in higher numbers of antigen-reactive effector CD4(+) and CD8(+) T cells producing granzyme and IFN-gamma, with lower levels of IL-10. Although immunodepletion studies indicate that both CD4 and CD8 T cells provide protection in the vaccinated mice, the contribution of CD4(+) T cells was significantly increased in mice primed with DNAp36 together with alphaGalCer. Notably 5 months after boosting, mice vaccinated with DNAp36 + alphaGalCer continued to show sustained and heightened T cell immune responses. Thus, heterologous prime-boost vaccination using alphaGalCer during priming is highly protective against murine cutaneous leishmaniasis, resulting in the heightened activation and development of CD4 and CD8 T cells (effector and memory T cells). PMID- 18286567 TI - IRAK-4 kinase activity is required for IRAK-4-dependent innate and adaptive immune responses. AB - Interleukin-1 receptor-associated kinase (IRAK)-4 is a serine-threonine kinase that plays an important role in innate and adaptive immune responses. While the requirement of IRAK-4 kinase activity has been studied in the context of IL-1R signaling, it is not clear whether IRAK-4 requires its kinase function for all of its roles in the immune system. IRAK-4 kinase-dead knock-in (IRAK-4KD/KD) mice were generated to further elucidate whether IRAK-4 kinase activity is required for IRAK-4 to induce cytokine production. IRAK-4KD/KD mice were impaired in their ability to produce cytokines in response to in vivo challenge with lipopolysaccharide (LPS), a potent TLR4 ligand. Cytokine production was also reduced in macrophages and dendritic cells from IRAK-4KD/KD mice in response to LPS and other TLR ligands. In addition, adaptive immune responses were impaired in IRAK-4KD/KD mice. Although in vitro T cell proliferation in response to TCR activation was unaffected in IRAK-4-deficient mice, in vivo T cell responses to lymphocytic choriomeningitits virus infection were significantly impaired in IRAK 4-knockout mice or mice expressing the kinase-dead mutant of IRAK-4. Collectively, these results indicate that IRAK-4 kinase activity is required for IRAK-4-dependent signaling in innate and adaptive immunity. PMID- 18286570 TI - Role of tripeptidyl peptidase II in MHC class I antigen processing - the end of controversies? AB - Peptide ligands presented by MHC class I molecules are generated in a cascade of proteolytic events starting with the proteasome in the cytosol and frequently terminating with trimming aminopeptidases in the endoplasmic reticulum. Several cytosolic proteases can carry out intermediate proteolytic steps between these start and endpoints. Among these, tripeptidyl peptidase II (TPP II), an exceptionally large homo-oligomeric protease, has been proposed to be involved in the generation of many or most MHC class I ligands by cleaving long precursor peptides. In this issue of the European Journal of Immunology, the effect of pharmacological or genetic TPP II inhibition on peptide loading of HLA-B27 and other HLA class I molecules is examined, and no evidence for a role of TPP II in this process is detected. Although further studies using more efficient inhibitors and focusing on HLA class I alleles such as HLA-A3 are warranted, these results, together with other recently published data, suggest that the role of TPP II in MHC class I processing may be much more limited than previously appreciated. PMID- 18286571 TI - IRAK4 in TLR/IL-1R signaling: possible clinical applications. AB - A member of the IL-1 receptor (IL-1R)-associated kinase (IRAK) family, IRAK4, has been shown to play an essential role in Toll-like receptor (TLR)-mediated signaling. IRAK4 kinase-inactive knockin mice have been shown to be completely resistant to LPS- and CpG-induced shock, due to impaired TLR-mediated induction of pro-inflammatory cytokines and chemokines. A reduction of LPS-, R848- and IL-1 mediated mRNA stability contributes to the reduced cytokine and chemokine production in bone marrow (BM)-derived macrophages from IRAK4 kinase-inactive knockin mice: however, not all of the TLR/IL-1R signaling events are ablated in IRAK4 kinase-inactive knockin mice. A paper in this issue of the European Journal of Immunology shows that, while JNK activation is significantly impaired, NF kappaB and IRF3 activation are retained in the absence of IRAK4 kinase activity. These residual TLR/IL-1R-induced signaling events allow the production of some cytokines and chemokines (including TNFalpha and CXCL1); at early times after the stimulation and induction of a group of TLR-mediated MyD88/IRAK4-independent genes in IRAK4 kinase-inactive knockin cells. Therefore, pharmacological blocking of IRAK4 kinase activity will retain some levels of host defence, while reducing the levels and duration of inflammatory responses, which should provide beneficial therapies for sepsis and chronic inflammatory diseases. PMID- 18286572 TI - Type I interferon-dependent and -independent expression of tripartite motif proteins in immune cells. AB - The tripartite motif (TRIM) proteins are important in a variety of cellular functions additional to anti-viral activity. We systematically analysed mRNA expression of representative TRIM molecules in mouse macrophages, myeloid and plasmacytoid dendritic cells, and a selection of CD4(+) T cell subsets. We defined four clusters of TRIM genes based on their selective expression in these cells. The first group of TRIM genes was preferentially expressed in CD4(+) T cells and contained the COS-FN3 motif previously shown to be involved in protein interactions. Additional TRIM genes were identified that showed up-regulation in macrophages and dendritic cells upon influenza virus infection in a type I IFN dependent manner, suggesting that they have anti-viral activity. In support of this notion, a subset of these TRIM molecules mapped to mouse chromosome 7, syntenic to human chromosome 11, where TRIM family members such as TRIM5, shown to have anti-viral activity, are localized. A distinct group of TRIM was constitutively expressed in plasmacytoid dendritic cells independently of viral infection or signalling through the type I IFN receptor. Our findings on expression and regulation of TRIM genes in cells of the immune system that have different effector functions in innate and adaptive immune responses, may provide leads for determining functions of this diverse family of molecules. PMID- 18286573 TI - Tripeptidyl peptidase II is dispensable for the generation of both proteasome dependent and proteasome-independent ligands of HLA-B27 and other class I molecules. AB - A significant fraction of the HLA-B27-bound peptide repertoire is resistant to proteasome inhibitors. The possible implication of tripeptidyl peptidase II (TPPII) in generating this subset was analyzed by quantifying the surface re expression of HLA-B*2705 after acid stripping in the presence of two TPPII inhibitors, butabindide and Ala-Ala-Phe-chloromethylketone. Neither decreased HLA B27 re-expression under conditions in which TPPII activity was largely inhibited. This was in contrast to a significant effect of the proteasome inhibitor epoxomicin. The failure of TPPII inhibition to decrease surface re-expression was not limited to HLA-B27, since it was also observed in several HLA-B27-negative cell lines, including Mel JuSo. Actually, HLA class I re-expression in Mel JuSo cells increased as a function of butabindide concentration, which is consistent with an involvement of TPPII in destroying HLA class I ligands. Inhibition of TPPII with small interfering RNA also failed to decrease the surface expression of HLA class I molecules on 143B cells. Our results indicate that TPPII is dispensable for the generation of proteasome-dependent HLA class I ligands and, without excluding its role in producing some individual epitopes, this enzyme is not involved to any quantitatively significant extent, in generating the proteasome-independent HLA-B27-bound peptide repertoire. PMID- 18286574 TI - Immunomodulation by herpesvirus U51A chemokine receptor via CCL5 and FOG-2 down regulation plus XCR1 and CCR7 mimicry in human leukocytes. AB - Human herpesvirus-6A (HHV-6A) betachemokine-receptor U51A binds inflammatory modulators CCL2, CCL5, CCL11, CCL7, and CCL13. This unique specificity overlaps that of human chemokine receptors CCR1, CCR2, CCR3, and CCR5. In model cell lines, expression leads to CCL5 down-regulation with both constitutive and inducible signaling. Here, immunomodulation pathways are investigated in human leukocytes permissive for infection. Constitutive signaling was shown using inositol phosphate assays and inducible calcium signaling by response to CCL2, CCL5 and CCL11. Constitutive signaling targets were examined using an immune response-related microarray and RT-PCR, showing down-regulation of CCL5 and FOG 2, a hematopoietic transcriptional repressor. By RT-PCR and siRNA reversion, CCL5 and FOG-2 were shown down-regulated, during peak U51A expression post infection. Two further active ligands, XCL1 and CCL19, were identified, making U51A competitor to their human receptors, XCR1 and CCR7, on T lymphocytes, NK and dendritic cells. Finally, U51A-expressing cell lines and infected ex vivo leukocytes, showed migration towards chemokine-gradients, and chemokine internalization. Consequently, U51A may affect virus dissemination or host transmission by chemotaxis of infected cells to sites of chemokine secretion specific for U51A (for example the lymph node or lung, by CCL19 or CCL11, respectively) and evade immune-effector cells by chemokine diversion and down regulation, affecting virus spread and inflammatory pathology. PMID- 18286575 TI - Polarity of major grooves explored by using an isosteric emissive nucleoside. PMID- 18286576 TI - Long-term occupational consequences of asthma in a large French cohort of male workers followed up for 5 years. AB - OBJECTIVES: The aim of this study was to describe the long-term occupational consequences of asthma in males of the ESTEV study, a French longitudinal cohort of working subjects aged 37-52 at inclusion. METHODS: Medical data, self perceived health status, sick leave, occupational social class and employment characteristics were recorded twice by occupational physicians in 1990 (12,233 subjects) and 1995 (10,608 subjects). Asthma was characterized as to its onset (childhood, i.e., before age 20 vs. adult) and to its past versus current status by the physician. RESULTS: Of the 398 asthmatics, the onset was before age 20 for 226 and the asthma status was classified as current for 159 subjects. Unemployment was not higher before baseline or during follow-up, in asthmatics as compared to non-asthmatics, despite a significantly higher prevalence of sick leave in the previous year among current asthmatics (38.4% vs. 27.0%, P = 0.005). Being a blue collar worker in 1990 is negatively related to childhood asthma but not to the current asthma status. In 1995, current adult-onset asthmatics had stopped working due to disability more frequently than never-asthmatics. CONCLUSION: Our findings suggest that the major consequence of asthma on employment status is a selective exclusion, observed in childhood asthmatics at the beginning of their working life and in current adult-onset asthmatics at the end of their working life. Past unemployment was shown not to be higher in working asthmatics. PMID- 18286579 TI - Early adolescents show enhanced acute cocaine-induced locomotor activity in comparison to late adolescent and adult rats. AB - Initiation of drug use during adolescence is associated with an increased probability to develop a drug addiction. The present study examined dose-response effects of cocaine (0, 5, 10, or 20 mg/kg, i.p.) on locomotor activity in early adolescent (postnatal day (PND) 35), late adolescent (PND 45), and young adults (PND 60) by measuring total distance moved (TDM) and frequency of start-stops. In response to 20 mg/kg cocaine, early adolescents showed the greatest cocaine induced increase in TDM in comparison to late adolescent and adult rats. At this same dose, early adolescents showed the greatest cocaine-induced attenuation of start-stops relative to older rats. Results suggest that early adolescents engage in more cocaine-induced locomotor activity and less stationary behavior indicating that early adolescents are more sensitive to locomotor activating effects of high dose cocaine than older rats. PMID- 18286580 TI - Understanding language and cognitive deficits in very low birth weight children. AB - Very-low-birth-weight infants are at much higher risk for cognitive and language delays but the nature of such deficits is not clearly understood. Given increasing rates of prematurity and infants born very-low-birth-weight, examination of mechanisms that underlie poorer developmental outcome is essential. We investigated language and cognitive abilities in very-low and normal birth-weight infants to determine whether performance differences were due to poorer global cognitive performance or to deficits in specific processing abilities. Thirty-two very-low and 32 normal birth-weight infants received visual and auditory-visual habituation recognition-memory tasks, and standardized language and cognitive assessments. Very-low-birth-weight infants performed more poorly on visual and auditory-visual habituation tasks and scored lower than controls on cognitive and language measures. These findings suggest that differences in language abilities in very-low-birth-weight children may be part of a global deficit that impacts many areas of cognitive functioning rather than a specific impairment in rapid auditory processing. PMID- 18286581 TI - Therapeutic effects of complex rearing or bFGF after perinatal frontal lesions. AB - We investigated the effects of an enriched environment and/or basic fibroblast growth factor (bFGF) on recovery from neonatal frontal injury in rats. Rats received medial frontal lesions, or sham surgery, on postnatal day (P) 2/3. In the first set of experiments (Experiments 1 and 2), rats were housed in enriched environments that consisted of a large enclosure with multiple objects (or standard housing) for 90 days beginning at weaning (P22) or in adulthood (P110). In Experiment 3, the rats either received 7 days of subcutaneous bFGF beginning on the day after surgery or bFGF plus enriched housing beginning at weaning. After the 90-day housing period, the animals were tested on a spatial navigation task and a skilled reaching task. Early lesions of the medial frontal cortex caused severe impairments in spatial learning but this deficit was markedly reduced with enriched housing, bFGF, or a combination of both, with the latter being most effective. The housing effects varied with age, however: the earlier the experience began, the better the outcome. Enriched housing increased dendritic length in cortical pyramidal neurons, an effect that was greater in the lesion than the control animals, and enriched housing reversed the lesion-induced decrease in spine density. Enriched environment increased the thickness of the cortical mantle in both lesion and controls whereas bFGF had no effect. Experience thus can affect functional and anatomical outcome after early brain injury but the effects vary with age at experience and may be facilitated by treatment with bFGF. PMID- 18286582 TI - Temporal association between food distribution and human caregiver presence and the development of affinity to humans in lambs. AB - The presence of the caregiver around feeding favors the development of a human animal relationship. To understand the underlying mechanism, we tested various temporal associations between food distribution and human presence: from an early age, a person was repeatedly present for 2 min just before milk distribution ("Forward"), during milk distribution ("Simultaneous"), and 20 min afterwards ("Delayed"). The "Control" group received no human contacts. During the treatments, "Forward" and "Delayed" lambs had more physical contacts with the person than "Simultaneous" lambs. When tested in unfamiliar environments, they stood longer near the person than did "Control" or "Simultaneous" lambs, which did not differ. Only "Forward" and "Delayed" lambs bleated when separated from the person. Fasting before testing had no effect. "Forward" and "Delayed" seemed to produce the same human-animal relationship, showing that this did not rely only on a conditioning process associating the caregiver with food. The caregiver may acquire properties for social support through other mechanisms (attachment and/or postingestive effects). PMID- 18286583 TI - A computer touch screen system and training procedure for use with primate infants: Results from pigtail monkeys (Macaca nemestrina). AB - Computerized cognitive and perceptual testing has resulted in many advances towards understanding adult brain-behavior relations across a variety of abilities and species. However, there has been little migration of this technology to the assessment of very young primate subjects. We describe a training procedure and software that was developed to teach infant monkeys to interact with a touch screen computer. Eighteen infant pigtail macaques began training at 90-postnatal days and five began at 180-postnatal days. All animals were trained to reliably touch a stimulus presented on a computer screen and no significant differences were found between the two age groups. The results demonstrate the feasibility of using computers to assess cognitive and perceptual abilities early in development. PMID- 18286584 TI - Physiological responses to social and physical challenges in children: Quantifying mechanisms supporting social engagement and mobilization behaviors. AB - Physiological response patterns to laboratory-based social and physical challenges were investigated in 37 typically-developing 3- to 5-year-old children. The study was conducted to determine whether the response profiles during each challenge were similar and whether individual differences in the response profiles to the challenges were correlated. Results demonstrated challenge specific physiological response strategies. In response to the social challenge, respiratory sinus arrhythmia and heart period increased and motor activity decreased. In contrast, in response to the physical challenge, respiratory sinus arrhythmia and heart period decreased and motor activity increased. Neither challenge reliably elicited changes in salivary cortisol. Only heart period responses were correlated between the challenges. PMID- 18286586 TI - Adrenocortical and behavioral attunement in parents with 1-year-old infants. AB - Sethre-Hofstad et al. [2002, Psychoneuroendocrinology 27:731-747] found that behaviorally well-attuned or sensitive parents showed better physiological attunement with their 2- to 4-year-old toddlers' adrenocortical responses to a potentially challenging task than less sensitive parents. In the present study we aimed to replicate this finding in a sample of 83 parents with 15-month-old infants. Parental and infant cortisol responses were assessed using saliva samples collected before and 21 min after the child's confrontation with a stranger and a moving robot. Infant behaviors reflecting distress/uncertainty during the stranger-robot session were rated from videotape. Parental sensitivity was observed during a parent-infant teaching episode. Our findings replicate those of Sethre-Hofstad et al. [2002, Psychoneuroendocrinology 27:731-747] by showing correlated parent-infant cortisol responses for sensitive parents but not for less sensitive parents. Furthermore, sensitive parents cortisol responses were associated with their children's distress/uncertainty during the stranger robot episode, whereas this was not true for less sensitive parents. Results indicate an important connection between behavior and physiology in parent-infant interactions that deserve more research. PMID- 18286585 TI - Return to school accompanied by changing associations between family ecology and cortisol. AB - This study examines everyday family life as a social regulator of child adrenocortical activity during the normative challenge of return to school. If positive family function facilitates child adaptation, we expected that mother child relationships following school entry would predict individual differences in evening cortisol, a context-sensitive marker for the response to concurrent demands. Among 28 children followed longitudinally, late in pre-kindergarten those living with single and/or employed mothers had higher evening cortisol. Yet early during the following school year, children with poorer mother-child relationships had higher evening cortisol. Cortisol awakening response, a comparatively stable marker of anticipated demands, was higher with maternal employment, single parents, and busier child schedules before school re-entry, and with maternal employment afterwards. We argue for a layered ecological approach to social regulation, recognizing that family structure, family functioning, and proximal features of everyday life within the family moderate child adrenocortical activity differently across contexts. PMID- 18286587 TI - The neuregulin 1 promoter polymorphism rs6994992 is not associated with chronic schizophrenia or neurocognition. AB - The neuregulin 1 (NRG1) promoter single nucleotide polymorphism (SNP) rs6994992 has shown association with decreased activation of frontal and temporal lobe regions, increased risk of psychosis, and decreased premorbid IQ. This SNP is part of a putative schizophrenia risk-associated haplotype and was associated with increased expression of the type IV transcript in postmortem tissue. We tested for association between rs6994992 and chronic schizophrenia by genotyping 738 cases from the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) and 733 matched controls. We further tested for associations with age at onset and baseline neurocognition in cases with schizophrenia reasoning that these phenotypes might yield results similar to those seen for premorbid IQ. Affection status was weakly associated with rs6994992 genotypes and trended towards association under a recessive model. This association did not survive correction for multiple comparisons and was in the opposite direction than has been reported. There was no association between rs6994992 and age at onset, an estimate of premorbid IQ, or neurocognition at study baseline. We were unable to replicate previous associations of rs6994992 with schizophrenia and, moreover, did not find significant associations with age of onset, an estimate of pre morbid IQ, or neurocognition. PMID- 18286588 TI - Evidence for potential relationship between SLC1A1 and a putative genetic linkage region on chromosome 14q to obsessive-compulsive disorder with compulsive hoarding. AB - Obsessive-compulsive disorder (OCD) is likely a disorder involving complex genetic transmission. This suggests that multiple genetic and environmental factors are involved in its etiology. This is complicated further by the probability of genetic heterogeneity for this phenotype. In this report, we describe a preliminary approach to deal with both complexities. SLC1A1, a glutamate transporter gene on chromosome 9p, was originally proposed to be related to OCD based on two linkage studies, and subsequently association of OCD to the gene has been replicated. Additionally, genetic linkage to a subtype of OCD, compulsive hoarding, has been reported on chromosome 14q. We hypothesized that both genomic regions contribute to OCD in some instances. Using the analytic program GENEFINDER we found that conditioning linkage on chromosome 14q to a marker adjacent to SLC1A1, reduced the size of the linkage region on chromosome 14q and provided evidence for interaction between the regions on chromosomes 9p and 14q. PMID- 18286589 TI - Cognitive dysfunction in mice deficient for TNF- and its receptors. AB - Recent evidence suggests a role for tumor necrosis factor alpha (TNF) in the functioning of the central nervous system (CNS). The aim of this work was to examine the effect of a deficiency of TNF (TNF(-/-)) and its main receptors (TNF R1(-/-) and TNF-R2(-/-)) on cognitive function. A standardized survey on cognition-like behavior assessing learning and retention, spatial learning/memory, cognitive flexibility, and learning effectiveness was used in B6.WT and B6.TNF gene targeted mice strains (B6.wild-type, B6.TNF(-/-), B6.TNF R1(-/-), B6.TNF-R2(-/-) mice). All studied mice strains demonstrated successful exploration and learning processes during the training phases of the tests, which made the specific cognition-like tests valid in these mice strains. In the specific cognition-like tests, the B6.TNF(-/-) mice demonstrated significantly poorer learning and retention in the novel object test compared to B6.WT, B6.TNF R1(-/-) and B6.TNF-R2(-/-) mice. In addition, spatial learning and learning effectiveness were significantly poorer in B6.TNF(-/-) mice compared to B6.WT mice. Moreover, the moderately impaired cognitive performance with similar degrees in B6.TNF-R1(-/-) or B6.TNF-R2(-/-) mice was generally better than in TNF(-/-) mice but also poorer than in B6.WT mice. While the absence of TNF was correlated with poor cognitive functioning, the deletion of both TNF-receptors was involved in partially reduced cognitive functioning. Low-levels of TNF under non-inflammatory immune conditions appear essential for normal cognitive function. TNF displays an interesting candidate gene for cognitive function. Translational research is required to investigate associations between genetic variants of TNF and cognitive function in healthy subjects and neuropsychiatric samples. PMID- 18286590 TI - Inhibition of mineralization by a calcium zirconium phosphate coating. AB - Bioactive ceramics used as coating materials combine the conductive properties of a bioceramic with the mechanical stability of the metal implant. We studied a calcium zirconium phosphate-containing coating material, FA-CZP [Ca(5)(PO(4))(3)F, CaZr(4)(PO(4))(6)], that is relatively insoluble in the biological milieu. The reaction of bone to this material was investigated histologically and histomorphometrically in an animal trial. Cylindrical Ti6Al4V specimens that had been coated with FA-CZP by plasma spraying were implanted in the femoral condyles of rabbits. The implants were left in place for 2, 4, 6, 12, and 14 weeks. FA-CZP led to impaired mineralization of the newly formed bone at the interface. Noncalcified osteoid was found throughout the whole study period. The layer seemed to become thicker with time. The mineralization disorder is evidently caused by zirconium ions. The presence of zirconium in the osteoid in contact with the implant was demonstrated by means of two different staining methods. PMID- 18286591 TI - Thin films of oriented collagen fibrils for cell motility studies. AB - Collagen films with oriented fibrils mimic tissues that have been remodeled by fibroblasts, which naturally tend to orient collagen fibrils in vivo. We have prepared thin films of ordered fibrils of collagen I, a major component of the extracellular matrix. The films were prepared by modifying a technique previously used to produce collagen I films for studies of cell morphology and intracellular signaling. By modifying the drying step, we were able to produce thin monolayers of collagen fibrils with consistent orientations over macroscopic (>100 microm) distances. We quantified the degree of orientation of the collagen fibrils using Fourier analysis of optical microscopy images. We also conducted experiments with vascular endothelial cells, and found that cell orientation and migration are well-correlated with fibril orientation. Using polarized cells, we showed oriented thin collagen film induces natural migration along the fibrils without using any sort of attractor. Taken together, these results demonstrate additional functionality and physiological relevance for a class of films being successfully applied in a variety of cell biology experiments. PMID- 18286592 TI - Fibroblast growth factor-2-apatite composite layers on titanium screw to reduce pin tract infection rate. AB - Fibroblast growth factor-2 (FGF-2)-apatite composite layers were formed on anodically oxidized titanium screws to improve bone-screw interface strength and to reduce pin tract infection rate through enhanced skin tissue healing in external fixation. A calcium-containing solution supplemented with FGF-2, a phosphate-containing solution, and a sodium bicarbonate solution were mixed at a Ca/P molar ratio of 2.0 to prepare a calcium phosphate solution supersaturated with respect to calcium phosphates. Screws were individually immersed in 10 mL of the calcium phosphate solution at 37 degrees C for 2 days. Low-crystalline apatite layers incorporating FGF-2 were formed on the screw surface at FGF-2 concentrations in the supersaturated calcium phosphate solution equal to or lower than 10 mug/mL. The amounts of FGF-2 immobilized on the screws ranged from 2.3- to 2.4-mug per screw. The immobilized FGF-2 retained biological activity, as demonstrated by NIH3T3 cell proliferation. Titanium screws with the composite layer were percutaneously implanted into the bilateral proximal tibial metaphyses in rabbits for 4 weeks. The titanium screws with the composite layer formed at the optimum FGF-2 concentration showed a significantly higher bone-screw interface strength and a lower pin tract infection rate than those without the composite layer: the extraction torque and infection rates were respectively 0.230 +/- 0.073 Nm and 43.8% for the screws with the composite layer, and 0.170 +/- 0.056 Nm and 93.8% for those without the composite layer. Therefore, titanium screws with the FGF-2-apatite composite layer are useful for improving bone-screw interface strength and infection resistance in external skeletal fixation. PMID- 18286593 TI - A biomechanical and histological analysis of standard versus hydroxyapatite coated pins for external fixation. AB - This sheep study was designed to make a comparative evaluation of two external fixation pin types each with and without hydroxyapatite (HA) coating. The two pins had different taper, pitch, and self drilling capabilities. Twenty Orthofix standard, self-tapping pins (group A), 20 Orthofix HA-coated, self-tapping pins (group B), 20 X-caliber, self-drilling, self-tapping pins (group C), and 20 X caliber HA-coated, self-drilling, self-tapping pins (group D) were selected. Four pins were implanted in the right femurs of 20 adult sheep that were euthanized at 6 weeks. Mean pin insertion torque was 2745 +/- 822 Nmm in group A, 2726 +/- 784 Nmm in group B, 2818 +/- 552 Nmm in group C, and 2657 +/- 732 Nmm in group D (ns). Mean pin extraction torque was 1567 +/- 541 Nmm in group A, 2524 +/- 838 Nmm in group B, 1650 +/- 650 Nmm in group C, and 2517 +/- 726 Nmm in group D. HA coated pins (group B and D) had a significantly greater mean pin extraction torque compared to similar uncoated pins (group A and C) (p < 0.0005). Histological analysis showed good osteointegration of the two coated pin types. This study shows that HA-coating is more important for optimal pin fixation than the particular combination of design parameters used in each pin type. PMID- 18286595 TI - A double-blind, parallel, multicenter comparison of L-acetylcarnitine with placebo on the attention deficit hyperactivity disorder in fragile X syndrome boys. AB - Attention deficit hyperactivity disorder (ADHD) is a frequent behavioral problem in young boys with fragile X syndrome (FXS), and its treatment is critical for improving social ability. The short-term efficacy of stimulant medications like methylphenidate (MPH) is well established in children with ADHD. FXS boys treated with MPH have improved attention span and socialization skills; however their mood becomes unstable at higher doses. Therefore, alternative pharmacological treatment of ADHD symptoms is desirable. A recent study showed that carnitine has a beneficial effect on the hyperactive-impulsive behavior in boys with ADHD without side effects. Our previous placebo-controlled trial indicated that L acetylcarnitine (LAC) reduces hyperactivity in FXS boys. The objective of this study was to determine the efficacy of LAC in a larger sample of FXS boys with ADHD. The study design was randomized, double blind placebo controlled, parallel, and multicenter (with eight centers involved in Italy, France, and Spain). Sixty three FXS males with ADHD (aged 6-13 years) were enrolled; 7 patients dropped out, 56 completed the one-year treatment, and 51 were included in the statistical analysis. Both groups improved their behavior, showing that psychosocial intervention has a significant therapeutic effect. However, we observed a stronger reduction of hyperactivity and improvement of social behavior in patients treated with LAC, compared with the placebo group, as determined by the Conners' Global Index Parents and the Vineland Adaptive Behavior Scale. Our results show that LAC (20-50 mg/kg/day) represents a safe alternative to the use of stimulant drugs for the treatment of ADHD in FXS children. PMID- 18286596 TI - A girl with fragile X premutation from sperm donation. AB - We present a girl with the fragile X premutation who obtained the premutation allele from donated sperm. Our patient has clinical characteristics of fragile X syndrome including emotional problems and neuropsychological difficulties presenting as learning disabilities. She is also at high risk for premature ovarian failure and low risk for the fragile X-associated tremor ataxia (FXTAS). We suggest fragile X DNA screening in gamete donor candidates to decrease the chance of fragile X involvement in their offspring. PMID- 18286597 TI - Genetic variation in the HTR1A gene and sudden infant death syndrome. PMID- 18286598 TI - Immunocytochemical localization of caveolin-3 in the synoviocytes of the rat temporomandibular joint during development. AB - Caveolins -- caveolin-1, -2, -3 (Cav1, 2, 3) -- are major components of caveolae, which have diverse functions. Our recent study on the temporomandibular joint (TMJ) revealed expressions of Cav1 and muscle-specific Cav3 in some synovial fibroblast-like type B cells with well-developed caveolae. However, the involvement of Cav3 expression in the differentiation and maturation of type B cells remains unclear. The present study therefore examined the chronological alterations in the localization of Cav3 in the synovial lining cells of the rat TMJ during postnatal development by immunocytochemical techniques. Observations showed immature type B cells possessed a few caveolae with Cav1 but lacked Cav3 protein at postnatal day 5 (P5). At P14, Cav3-immunopositive type B cells first appeared in the synovial lining layer. They increased in number and immunointensity from P14 to P21 as occlusion became active. In immunoelectron microscopy and double immunolabeling with heat shock protein 25 (Hsp25) and Cav3, coexpressed type B cells developed rough endoplasmic reticulum and numerous caveolae, while the Cav3-immunonegative type B cell with Hsp25 immunoreaction possessed few of these. Results suggest that Cav3 expression, which is closely related to added functional stimuli, reflects the differentiation of the type B synoviocytes. PMID- 18286599 TI - Unipolar depression and hippocampal volume: impact of DNA sequence variants of the glucocorticoid receptor gene. AB - Glucocorticoid receptor (GR) plays a major role in regulation of the hypothalamic pituitary-adrenocortical (HPA) system; HPA dysregulation represents the most consistent biological pattern of depression. Multiple functional polymorphisms are known for the GR gene, which might influence the development of unipolar depression. Previous studies reported associations to some variants in this gene but not consistently so. We investigated seven genetic polymorphisms in the GR gene (NR3C1) located in the putative promoter, exon 2 and intron 2 region. Study populations were 322 German inpatients with recurrent unipolar depression, and 298 German controls recruited from the general population. The relationships between intermediate phenotypes (hippocampal and amygdala volumes) and NR3C1 DNA sequence variants were additionally explored in a subpopulation of patients. We found association between the diagnosis of depression and DNA sequence variants in intron 2 as well as in the 5' region of the NR3C1 gene but not for the previously studied exon 2 and putative promoter variants (global test after control of multiple testing, P = 0.02). In patients, diagnosis-related alleles were also associated to hippocampal volume reduction and amygdala volume variation. Unipolar depression is associated with DNA variants of the NR3C1 gene in our population. Neurobiological underpinnings of depression as volumetric reductions of the hippocampus may also be mediated by variants in this gene. PMID- 18286600 TI - Twenty-four-hour shift work, depressive symptoms, and job dissatisfaction among Japanese firefighters. AB - BACKGROUND: The influence of a 24-hr shift-work burden on firefighters' mental health has not been fully investigated. The purpose of this study is to clarify the relationships between specific workload items among firefighters engaged in 24-hr shift work and job stress as estimated by the generic job-stress questionnaire on depressive symptoms and job dissatisfaction from the National Institute for Occupational Safety and Health (NIOSH). METHOD: The subjects were 1,301 firefighters. The questionnaire covered age, gender, job type, job class, marital status, smoking and drinking habits, number of attendances, turnout time, extra work hours, average nap-time, the Center for Epidemiologic Studies Depression Scale (CES-D), and questions from the NIOSH generic job-stress questionnaire, including those on job satisfaction. RESULTS: In multivariate logistic regression analyses, a model that included all variables except the measures from the NIOSH generic job-stress questionnaire showed that shorter nap time had significant higher odds ratios (ORs) for depressive symptoms and job dissatisfaction, but the significances disappeared in a fully adjusted model. In fully adjusted logistic regression analyses, low quantitative workload, low variance in workload, high intra- and intergroup conflict, low social support from a supervisor, high role conflict and ambiguity, and low self-esteem had significant higher ORs for depressive symptoms and/or job dissatisfaction. CONCLUSIONS: Amount of workload, variance in workload, intra- and intergroup conflict, social support from a supervisor, role conflict and ambiguity, and self esteem were significantly related to depressive symptoms and/or job dissatisfaction among Japanese firefighters. Moreover, inadequate nap-time may affect their mental health. PMID- 18286601 TI - Bone formation and resorption in patients after implantation of beta-tricalcium phosphate blocks with 60% and 75% porosity in opening-wedge high tibial osteotomy. AB - Most of the implanted porous beta-tricalcium phosphate (beta-TCP) can be resorbed. However, beta-TCP block with 75% porosity is inadequate for weight bearing sites until bone incorporation occurs. Thus, the authors have recently developed beta-TCP block with 60% porosity, which is approximately sevenfold greater in terms of compressive strength than that of beta-TCP with 75% porosity. The authors investigated bone formation and resorption of beta-TCP after implantation in patients of beta-TCP blocks with two different porosities. From May 2003 to November 2004, medial opening high tibial osteotomy was performed in 25 patients with a mean age of 66 years. The opened defect was fixed with a Puddu plate. Then 6-8 cm(3) of beta-TCP block with 75% porosity was used to fill the cancellous bone defect, except on the medial side where 2.83-3.18 cm(3) of wedge shaped beta-TCP block with 60% porosity was implanted. At least 2 years after surgery, the 25 patients had no correction loss, and bone formation was noted in all cases. Complete or nearly complete resorption of beta-TCP with 60 and 75% porosity was obtained within 3.5 years. Thirteen biopsy samples obtained from the 60% porosity implantation sites showed good lamellar bone formation, and the percentage of beta-TCP remaining relative to the newly formed bone plus beta-TCP ranged from 0.3 to 14.5%, with a mean of 6.7%. The authors suspect that mechanical stress loading to the medial side of the tibia facilitated bone formation and resorption of beta-TCP with 60% porosity. PMID- 18286602 TI - Mechanical behavior of hydroxyapatite biomaterials: an experimentally validated micromechanical model for elasticity and strength. AB - Hydroxyapatite (HA) biomaterials production has been a major field in biomaterials science and biomechanical engineering. As concerns prediction of their stiffness and strength, we propose to go beyond statistical correlations with porosity or empirical structure-property relationships, as to resolve the material-immanent microstructures governing the overall mechanical behavior. The macroscopic mechanical properties are estimated from the microstructures of the materials and their composition, in a homogenization process based on continuum micromechanics. Thereby, biomaterials are envisioned as porous polycrystals consisting of HA needles and spherical pores. Validation of respective micromechanical models relies on two independent experimental sets: biomaterial specific macroscopic (homogenized) stiffness and uniaxial (tensile and compressive) strength predicted from biomaterial-specific porosities, on the basis of biomaterial-independent ("universal") elastic and strength properties of HA, are compared with corresponding biomaterial-specific experimentally determined (acoustic and mechanical) stiffness and strength values. The good agreement between model predictions and the corresponding experiments underlines the potential of micromechanical modeling in improving biomaterial design, through optimization of key parameters such as porosities or geometries of microstructures, in order to reach the desired values for biomaterial stiffness or strength. PMID- 18286603 TI - Effect of adding sodium hexametaphosphate liquefier on basic properties of calcium phosphate cements. AB - Sodium hexametaphosphate (Na-HMP) is a common liquefying agent widely used in the ceramics industry for modifying the rheological behavior of ceramic slurries. The objective of this study was to investigate the influence of Na-HMP on several properties of calcium phosphate cements (CPCs). Various types of CPCs were prepared by mixing the same powder, namely, a mixture of tetracalcium phosphate and dicalcium phosphate anhydrate, with various liquids, namely, distilled water, Na(2)HPO(4) solutions, and Na-HMP solutions. The setting time, mechanical strength, rheological properties, and injectability of the cement pastes were examined in this work. Also, X-ray diffractometry (XRD) and scanning electron microscopy (SEM) techniques were employed for phase analysis and morphological evaluations, respectively. The results showed that, compared to CPCs made with water and Na(2)HPO(4) solutions, the CPC pastes made with Na-HMP solutions had improved stability and injectability but prolonged setting times. The XRD and SEM studies showed that Na-HMP inhibited the growth of apatite crystals during soaking of the cements in Ringer's solution. Thus, the CPC made with Na-HMP solution had lower compressive strength than those made with water or Na(2)HPO(4) solutions. Although Na-HMP could improve the injectability and stability of the CPC paste, it impaired other basic properties of the cement. Thus, it is not an appropriate liquefier additive. PMID- 18286604 TI - Side effects of a non-peroxide-based home bleaching agent on dental enamel. AB - Changes in the chemistry and structure of enamel due to a non-peroxide-based home bleaching product (Rapid White) were studied in vitro using attenuated total reflectance-infrared spectroscopy, Raman spectroscopy, electron probe microanalysis, flame atomic absorption spectroscopy, and total reflection X-ray fluorescence. The results revealed that the citric-acid-containing gel-like component of the bleaching system substantially impacts on the dental hard tissue. Enamel is affected on several levels: (i) the organic component is removed from superficial and deeper enamel layers and remnants of the bleaching gel are embedded in the emptied voids; (ii) cracks and chemical inhomogeneities with respect to Ca and P occur on the surface; and (iii) within a submicron layer of enamel, the Ca-O bond strength in apatite decreases, thus enhancing calcium leakage from the bleached enamel hard tissue. PMID- 18286605 TI - Comparison of 2D fiber network orientation measurement methods. AB - The mechanical properties of tissues, tissue analogs, and biomaterials are dependent on their underlying microstructure. As such, many mechanical models incorporate some aspect of microstructure, but a robust protocol for characterizing fiber architecture remains a challenge. A number of image-based methods, including mean intercept length (MIL), line fraction deviation (LFD), and Fourier transform methods (FTM), have been applied to microstructural images to describe material heterogeneity and orientation, but a performance comparison, particularly for fiber networks, has not been conducted. In this study, we constructed 40 two-dimensional test images composed of simulated fiber networks varying in fiber number, orientation, and anisotropy index. We assessed the accuracy of each method in measuring principal direction (theta) and anisotropy index (alpha). FTM proved to be the superior method because it was more reliable in measurement accuracy (Deltatheta = 2.95 degrees +/- 6.72 degrees , Deltaalpha = 0.03 +/- 0.02), faster in execution time, and flexible in its application. MIL (Deltatheta = 6.23 degrees +/- 10.68 degrees , Deltaalpha = 0.08 +/- 0.06) was not significantly less accurate than FTM but was much slower. LFD (Deltatheta = 9.97 degrees +/- 11.82 degrees , Deltaalpha = 0.24 +/- 0.13) consistently underperformed. FTM results agreed qualitatively with fibrin gel SEM micrographs, suggesting that FTM can be used to obtain image-based statistical measurements of microstructure. PMID- 18286606 TI - Preparation and in vitro characterization of novel bioactive glass ceramic nanoparticles. AB - SiO(2)-CaO-P(2)O(5) ternary bioactive glass ceramic (BGC) nanoparticles with different compositions were prepared via a three-step sol-gel method. Polyethylene glycol was selected to be used as the surfactant to improve the dispersion of the nanoparticles. The morphology and composition of these BGC nanoparticles were observed by ESEM and EDX. All the BGC particles obtained in this method were about 20 nm in diameter. XRD analysis demonstrated that the different compositions can result in very different crystallinities for the BGC nanoparticles. Bioactivity tests in simulated body fluid solution (SBF), and degradability in phosphate buffer solution (PBS), were performed in vitro. SEM, EDX, and XRD were employed to monitor the surface variation of neat poly(L-lactic acid), PLLA, foam and PLLA/BGC porous scaffolds during incubation. The BGC nanoparticles with lower phosphorous and relative higher silicon content exhibited enhanced mineralization capability in SBF and a higher solubility in PBS medium. Such novel nanoparticles may have potential to be used in different biomedical applications, including tissue engineering or the orthopedic field. PMID- 18286607 TI - Volumetric neuroimaging of the atlantic white-sided dolphin (Lagenorhynchus acutus) brain from in situ magnetic resonance images. AB - The structure and development of the brain are extremely difficult to study in free-ranging marine mammals. Here, we report measurements of total white matter (WM), total gray matter (GM), cerebellum (WM and GM), hippocampus, and corpus callosum made from magnetic resonance (MR) images of fresh, postmortem brains of the Atlantic white-sided dolphin (Lagenorhynchus acutus) imaged in situ (i.e., the brain intact within the skull, with the head still attached to the body). WM:GM volume ratios of the entire brain increased from fetus to adult, illustrating the increase in myelination during ontogeny. The cerebellum (WM and GM combined) of subadult and adult dolphins ranged from 13.8 to 15.0% of total brain size, much larger than that of primates. The corpus callosum mid-sagittal area to brain mass ratios (CCA/BM) ranged from 0.088 to 0.137, smaller than in most mammals. Dolphin hippocampal volumes were smaller than those of carnivores, ungulates, and humans, consistent with previous qualitative results assessed from histological studies of the bottlenose dolphin brain. These quantitative measurements of white matter, gray matter, corpus callosum, and hippocampus are the first to be determined from MR images for any cetacean species. We establish here an approach for accurately determining the size of brain structures from in situ MR images of stranded, dead dolphins. This approach can be used not only for comparative and developmental studies of marine mammal brains but also for investigation of the potential impacts of natural and anthropogenic chemicals on neurodevelopment and neuroanatomy in exposed marine mammal populations. PMID- 18286608 TI - Knee posture predicted from subchondral apparent density in the distal femur: an experimental validation. AB - Spatial patterning in the apparent density of subchondral bone can be used to discriminate between species that differ in their joint loading conditions. This study provides an experimental test of two hypotheses that relate aspects of subchondral apparent density patterns to joint loading conditions. First, the region of maximum subchondral apparent density (RMD) will correspond to differences in joint posture at the time of peak locomotor loads; and second, differences in maximum density between individuals will correspond to differences in exercise level. These hypotheses were tested using three age-matched samples of juvenile sheep. Two groups of five sheep were exercised, at moderate walking speeds, twice daily for 45 days on a treadmill with either a 0% or 15% grade. The remaining sheep were not exercised. Sheep walking on the inclined treadmill used more flexed knee postures than those in the level walking group at the time of peak vertical ground reaction forces. Kinematic measurements of knee posture were compared with knee postures estimated from the spatial position of the RMD on the medial femoral condyle. Our results show that the difference in the position of the RMD between the incline and level walking groups corresponded to the difference in knee postures obtained kinematically; however, exercised and nonexercised sheep did not differ in the magnitude of apparent density. These results suggest that patterns of subchondral apparent density are good indicators of the experimental modifications in joint posture during locomotion and may, therefore, be used to investigate differences between species in habitual joint loading. PMID- 18286609 TI - Evaluation of tissue eosinophilia in the pouch and afferent limb in patients with restorative proctocolectomy. AB - BACKGROUND: Although tissue eosinophilia in mucosal biopsy specimens from the ileal pouch in patients treated with restorative proctocolectomy is frequently seen, its clinical significance has not been investigated. The aim of this study was to assess whether tissue eosinophilia was associated with disease status of ileal pouches. METHODS: Hematoxylin and eosin slides of pouch and afferent limb biopsy specimens from 106 patients randomly selected from the Pouchitis Database were evaluated by 2 gastrointestinal pathologists. Of 106 patients, 81 had corresponding mucosal biopsy specimens of the afferent limb that were available for review. Tissue eosinophil infiltration was evaluated in a semiquantitative fashion with scores ranging from 0-3. Univariate and multivariate analyses were performed to assess the association between eosinophil scores and demographic, clinical, endoscopic, and histologic features. RESULTS: Multivariate analyses showed that tissue eosinophilia of the pouch and afferent limb was not associated with chronic inflammatory conditions of the pouch and the presence of concurrent autoimmune-mediated disorders. Tissue eosinophil score of the pouch was significantly higher than that in the corresponding afferent limb in the same patient population (P = 0.043). A high tissue eosinophil score in the afferent limb was associated with non-use nonsteroidal antiinflammatory drug use (odds ratio = 3.5; 95% confidence interval [CI]: 1.2, 10.4) and high endoscopic inflammation scores in the afferent limb (odds ratio = 1.6; 95% CI: 1.1, 2.2). Similar associations were not found in pouch biopsy specimens. CONCLUSIONS: Tissue eosinophilia in the pouch was more prominent than that in the afferent limb in patients with restorative proctocolectomy. Tissue eosinophilia in the pouch and afferent limb appeared to be associated with different risk factors. These findings suggest that luminal factors in different topographical locations of the pouch may contribute to eosinophil-mediated inflammation at these sites. PMID- 18286611 TI - Importance of diagnosis of concomitant cytomegalovirus infection in patients with intestinal Behcet's disease. PMID- 18286610 TI - Activated Notch1 maintains the phenotype of radial glial cells and promotes their adhesion to laminin by upregulating nidogen. AB - Radial glia are neural stem cells that exist only transiently during central nervous system (CNS) development, where they serve as scaffolds for neuronal migration. Their instability makes them difficult to study, and therefore we have isolated stabilized radial glial clones from E14.5 cortical progenitors (e.g., L2.3) after expression of v-myc. Activated Notch1 intracellular region (actNotch1) promotes radial glia in the embryonic mouse forebrain (Gaiano et al., (2000), and when it was introduced into E14.5 cortical progenitors or radial glial clone L2.3, the cells exhibited enhanced radial morphology and increased expression of the radial glial marker BLBP. A representative clone of L2.3 cells expressing actNotch1 called NL2.3-4 migrated more extensively than L2.3 cells in culture and in white matter of the adult rat spinal cord. Microarray and RT-PCR comparisons of mRNAs expressed in these closely related clones showed extensive similarities, but differed significantly for certain mRNAs including several cell adhesion molecules. Cell adhesion assays demonstrated significantly enhanced adhesion to laminin of NL2.3-4 by comparison to L2.3 cells. The laminin binding protein nidogen was the most highly induced adhesion molecule in NL2.3-4, and immunological analyses indicated that radial glia synthesize and secrete nidogen. Adhesion of NL2.3-4 cells to laminin was inhibited by anti-nidogen antibodies and required the nidogen binding region in laminin, indicating that nidogen promotes cell adhesion to laminin. The combined results indicate that persistent expression of activated Notch1 maintains the phenotype of radial glial cells, inhibits their differentiation, and promotes their adhesion and migration on a laminin/nidogen complex. PMID- 18286612 TI - Steroid hormone receptor expression and function in microglia. AB - Steroid hormones such as glucocorticoids and estrogens are well-known regulators of peripheral immune responses and also show anti-inflammatory properties in the brain. However, the expression of steroid hormone receptors in microglia, the pivotal immune cell that coordinates the brain inflammatory response, is still controversial. Here we use real time RT-PCR to show that microglia, isolated from adult fms-EGFP mice by FACS, express glucocorticoid receptor (GR), mineralocorticoid receptor (MR), and estrogen receptor alpha (ERalpha). GR was the most abundant steroid hormone receptor transcript in microglia. The presence of GR and ERalpha immunoreactivity was further confirmed in vivo at the ultrastructural level. To understand the role of steroid hormone receptors during the inflammation process, we evaluated the expression of steroid hormone receptors after inflammatory challenge and found a significant down-regulation of GR, MR, and ERalpha in microglia. Finally, we tested the immunomodulatory properties of estrogens and glucocorticoids. Estradiol benzoate did not have any significant impact on the inflammatory profile of ex vivo sorted microglia, either in resting conditions or after challenge. Furthermore, corticosterone was a more consistent anti-inflammatory agent than 17beta-estradiol in vitro. Our results support the hypothesis that adult microglia are a direct target of steroid hormones and that glucocorticoids, through the predominant expression of GR and MR, are the primary steroid hormone regulators of microglial inflammatory activity. The down-regulation of steroid hormone receptors after LPS challenge may serve as a prerequisite to suppressing the anti-inflammatory actions of endogenous steroid hormones on the immune system, and contribute to a sustained activation of microglia. PMID- 18286613 TI - Renal intratubular crystals and hyaluronan staining occur in stone formers with bypass surgery but not with idiopathic calcium oxalate stones. AB - Whether idiopathic calcium oxalate (CaOx) stone formers form inner medullary collecting duct (IMCD) crystal deposits bears on pathogenetic mechanisms of stone formation. In prior work, using light and transmission electron microscopy, we have found no IMCD crystal deposits. Here, we searched serial sections of papillary biopsies from a prior study of 15 idiopathic calcium oxalate stone formers, 4 intestinal bypass patients with CaOx stones, and 4 non-stone-forming subjects, and biopsies from an additional hitherto unreported 15 idiopathic calcium oxalate stone formers and 1 bypass patient using polarized light oil immersion optics, for deposits overlooked in our original study. We found no IMCD deposits in any of 1,500 serial sections from the 30 idiopathic calcium oxalate stone formers, nor in 87 additional sections from a frozen idiopathic calcium oxalate stone former biopsy sample processed without exposure to aqueous solutions. Among 4 of the 5 bypass patients but in none of the 30 idiopathic calcium oxalate stone formers or 4 normal stone formers, we found tiny birefringent thin crystalline overlays on scattered IMCD cell membranes. We also found IMCD lumen deposits in two bypass patients that contained mixed birefringent and nonbirefringent crystals, presumably CaOx and apatite. In the bypass patients, we observed focal apical IMCD cell hyaluronan staining, which was absent in idiopathic calcium oxalate stone formers. The absence of any IMCD deposits in 1,500 serial sections of biopsies from 30 idiopathic calcium oxalate stone formers allows us to place the upper limit on the probability of their occurrence at approximately 0.002 and place the lower limit of their size at the resolution of the optics (<0.2 mu). The tiny deposits in bypass patients may be the initial crystal lesion. PMID- 18286614 TI - Digit ratios in green anolis lizards (Anolis carolinensis). AB - The development of tetrapod digits is directed by the Homeobox (Hox) genes. The expression of Hox genes is influenced by exposure to endogenous sex steroids during development so that prenatal exposure to estrogens and androgens positively influences the lengths of digits 2 (2D) and 4 (4D), respectively. Because of this, Manning (2002) predicted that male tetrapods should have smaller 2D:4D than that of females because males are exposed to higher levels of androgens during development. We measured digits in green anolis lizards (Anolis carolinensis) to (a) test Manning's prediction; (b) compare our results with those of other studies of digit ratios in reptiles and birds to test the phylogenetic constraint hypothesis, which argues that the digit ratios of reptiles and birds should be more similar to each other than either taxa is to mammals because the patterns have been conserved in living diapsids; and (c) compare our results with those of another study of digit ratio in anoles (Chang et al., 2006). We did not detect evidence of sexual dimorphism in 2D:4D. Our results did not support either Manning's prediction or the phylogenetic constraint hypothesis. Furthermore, our results did not match those found in Chang et al. (2006), suggesting that digit ratios in anolis lizards may not be reliable indicators of prenatal exposure to hormones. PMID- 18286616 TI - Immunohistochemical and morphometric study of pituitary pars distalis thyrotrophs of male viscacha (Lagostomus maximus maximus): seasonal variations and effect of melatonin and castration. AB - The physiology and behavior of viscacha vary along the year according to the modifications of environmental signals such as the photoperiod length, temperature, rainfall pattern, food composition, and social interactions. The pituitary pars distalis thyrotrophs (TSH cells) of male viscacha were immunohistochemically identified, and the morphometric parameters: percentage of immunopositive area (% IA), cell percentage in pars distalis (% PDC), number of cells per reference area (no. cells/RA), and major cellular and nuclear diameters were analyzed. Three different groups of adult male viscachas were used: (1) captured in their natural habitat during the year, (2) melatonin-administered, and (3) castrated. The thyrotrophs were localized in the ventromedial sector, mainly in the pars distalis cephalic extreme. They were oval or pyramidal in shape, and their immunostaining intensity was heterogeneous. The % IA, % PDC, and no. cells/RA exhibited a significant decrease in June-July (winter, gonadal regression period) in relation to February-March (summer-early autumn, reproductive period), and they were recovered in August-September (later winter early spring, gonadal recovery period). No morphometric variations of TSH cells were observed in melatonin-treated animals, whereas a decrease of the % IA, % PDC, and no. cells/RA was observed in castrated animals in relation to the intact animals. Our results show TSH cell morphometric variations during the year in agreement with the animal's different physiological conditions during the reproductive cycle, and probably in response to the environmental signals changes. Melatonin does not have a direct effect on the TSH cells. However, castration modifies some thyrotroph morphometric parameters, reinforcing the hypothesis that androgens affect the cells activity. PMID- 18286615 TI - Optimization of volumetric computed tomography for skeletal analysis of model genetic organisms. AB - Forward and reverse genetics now allow researchers to understand embryonic and postnatal gene function in a broad range of species. Although some genetic mutations cause obvious morphological change, other mutations can be more subtle and, without adequate observation and quantification, might be overlooked. For the increasing number of genetic model organisms examined by the growing field of phenomics, standardized but sensitive methods for quantitative analysis need to be incorporated into routine practice to effectively acquire and analyze ever increasing quantities of phenotypic data. In this study, we present platform independent parameters for the use of microscopic x-ray computed tomography (microCT) for phenotyping species-specific skeletal morphology of a variety of different genetic model organisms. We show that microCT is suitable for phenotypic characterization for prenatal and postnatal specimens across multiple species. PMID- 18286617 TI - Genotoxicity and cytotoxicity in multiple organs induced by titanium miniplates in Wistar rats. AB - Internal fixture materials are currently used as metallic biomaterials in rehabilitation of human body. Nevertheless, metal release due to corrosion phenomena appears to play a crucial role for human health. Thus, the goal of the present study was to evaluate whether liver, kidney, and lung are particularly sensitive organs for DNA damaging and cytotoxicity following implantation of internal fixture materials composed by titanium alloy in vivo. A total of 18 rats underwent surgical titanium miniplates in their tibias, being randomly distributed into three groups: 30 days, 90 days, and 180 days after implantation. A total of six animals served as negative control (animals that not received any implant). After experimental design, the lung, liver, and kidney were removed for histopatological and genotoxic analysis as depicted by H.E. stain and single cell gel (comet) assay, respectively. No significant statistically differences (p > 0.05) for DNA damaging were found to all experimental groups when compared to negative control for all organs evaluated. In addition, no remarkable morphological alterations were detected under histopathological analysis. Taken together, such results suggest that titanium miniplates are neither able to induce DNA damage in multiple organs nor to cause some abnormalities in lung, liver, and kidney. PMID- 18286618 TI - Antibacterial properties of Ag (or Pt)-containing calcium phosphate coatings formed by micro-arc oxidation. AB - Silver (or platinum)-containing calcium phosphate (hydroxyapatite (HA) and tricalcium phosphate (alpha-TCP)) coatings on titanium substrates were formed by micro-arc oxidation (MAO) and their in vitro antibacterial activity and in vitro cytotoxicity were evaluated. MAO was performed in an electrolytic solution containing beta-glycerophosphate disodium salt pentahydrate (beta-GP) and calcium acetate monohydrate (CA), and Ag and Pt were introduced in the form of AgNO(3) (or CH(3)COOAg) and H(2)PtCl(6), respectively. The MG63 and human osteosarcoma (HOS) cell lines were used to investigate the proliferation and differentiation behavior of the cells, respectively, whereas two strains of bacteria, Staphylococcus aureus and Escherichia coli, were used to evaluate the antibacterial activity of the coatings. The phase, morphology, and Ag content of the coatings were strongly dependent on the applied voltage and Ag precursor concentration. HA and alpha-TCP phases were detected in the coatings oxidized above 400 V and the presence of Ag was confirmed by EDS. While the coatings with a high content of Ag were cytotoxic and those obtained in the Pt-containing electrolyte had no apparent antibacterial activity, the calcium phosphate coatings obtained in the low Ag concentration electrolyte exhibited in vitro antibacterial activity but no cytotoxicity. Thus, biocompatible calcium phosphate coatings on Ti implants with antibacterial activity can be achieved by one-step MAO. PMID- 18286619 TI - Boronate-containing polymer brushes: characterization, interaction with saccharides and mammalian cancer cells. AB - Boronate-containing polymer brushes were synthesized by free radical copolymerization of N,N-dimethylacrylamide (DMAA) and N-acryloyl-m-phenylboronic acid (NAAPBA) (9:1) on the surface of 3-mercaptopropyl-silylated glass plates and capillaries. The brushes were characterized with time-of-flight secondary ion mass-spectrometry (ToF SIMS), atomic force microscopy and contact angle measurements. Fructose caused a well-expressed drop spreading on the surface of copolymer-grafted glass, due to the strong interaction with the boronate groups. Sedimentation of murine hybridoma cells M2139 or human myeloid leukemia cells KG1 onto the DMAA-NAAPBA copolymer-grafted glass plates from 10 mM phosphate buffer solution (pH 8.0) resulted in the cell adhesion. The adhered M2139 and KG1 cells could be quantitatively detached from the grafted plates with 0.1 M fructose, which competed with cell surface carbohydrates for binding to the boronates. Evaluation of the binding strength between M2139 cells and the copolymer brush was performed by exposure of the adhered cells to a shear stress. Detachment of a fraction of 18% of the adhered M2139 cells was obtained at a shear force of 1400 2800 pN/cell generated by the running phosphate buffer (pH 8.0), whereas the remaining adhered cells (70%) could be detached with 0.1 M fructose dissolved in the same buffer. Possible applications of the boronate-containing polymer brushes to affinity cell separation can be based upon the facile recovery of the attached cells. PMID- 18286620 TI - Method for predicting sorption of small drug molecules onto polylactide. AB - A method has been developed to predict the percent sorption of small drug molecules onto polylactide (PLA). In this method, molecular modeling is first used to calculate the interaction energy between the adsorbate and PLA. Alternatively, the adsorbate-PLA interaction energy can be calculated by summing the adsorbate functional group contributions to the interaction energy. These functional group contributions have been found in this study. The adsorbate-PLA interaction energy is then substituted into a linear equation to predict the natural log of the sorption equilibrium constant, and the percent sorption is obtained from this value. This linear equation has been developed based on experimental sorption data for dyes on PLA. The predicted percent sorption of two small drugs on PLA has been verified with experimental data. Our method possibly could be used to select appropriate drugs for use in controlled release studies involving PLA. This method also could be used to calculate the functional group contributions to the interaction energies between other drugs and polymers and to predict their percent sorption onto those polymers, which could be used to select drugs suitable for controlled release studies involving those polymers. PMID- 18286621 TI - Enhancement of the biological activity of BMP-2 by synthetic dextran derivatives. AB - In the present study, we explored the binding capacity of synthetic heparin-like dextran derivatives to recombinant human bone morphogenetic protein 2 (BMP-2), a heparin-binding osteoinductive growth factor. Affinity electrophoresis analysis provided evidence that carboxymethylated dextran polymers grafted with high amounts of benzylamide groups (named DMCB) interact with BMP-2. The capability of such polysaccharides to potentiate the growth factor biological activity was then investigated. In vitro, DMCB dose-dependently promoted osteoblast differentiation induced by BMP-2 in C2C12 myoblasts more efficiently than heparin. A screening study provided evidence that the potentiating effects of the dextran derivatives on the BMP-2-induced alkaline phosphatase activity improved with their benzylamide groups content and, therefore, with their affinity for the growth factor. The biological activity of BMP-2 was monitored in the culture medium after 6 days using C2C12 cells (containing a BMP sensitive luciferase reporter gene). Like heparin, DMCB sustained the biological activity of the growth factor; this result suggests that the formation of the BMP-2/DMCB complex may protect the protein from being inactivated. In rats in vivo, DMCB also stimulated ectopic calcification mediated by BMP-2. These data indicate that dextran-based polysaccharides prolong the half-life of the growth factor and promote its biological activity. PMID- 18286622 TI - Endothelial cell migration in human plasma is enhanced by a narrow range of added sphingosine 1-phosphate: implications for biomaterials design. AB - Sphingosine 1-phosphate (S1P) promotes endothelial cell migration in vitro and may potentially impact the endothelialization of implanted biomaterials. However, the effects of S1P on endothelial cells (EC) in flowing blood could be negligible due to preactivation of signaling cascades. We previously developed biomaterials that release S1P and wished to determine through in vitro experiments the extent to which EC respond to S1P added to human platelet poor plasma. We found that addition of 200 nM S1P to platelet poor plasma significantly increased cell migration in two migration models. A lower concentration of S1P added to plasma (100 nM) did not increase endothelial cell migration rates, while the cell migration response was saturated above 200 nM S1P. Expression of the main S1P receptor in EC, S1P(1), was elevated in plasma compared to low serum medium, but addition of VEGF or fluid flow elicited a further increase in S1P(1) mRNA, consistent with the synergistic effects observed between S1P, VEGF, and fluid flow. Thus, sustained delivery of S1P from biomaterials might only enhance endothelial cell migration if the concentration of S1P at the surface of the material stimulated adjacent EC to the same extent as approximately 200 nM S1P added to plasma. PMID- 18286623 TI - Composition of corrosion layers on a magnesium rare-earth alloy in simulated body fluids. AB - In this study, the composition of corrosion product layers on a magnesium rare earth alloy in simulated body fluid (m-SBF) containing albumin in physiological concentration is examined. The time dependence of the composition of the layer is studied. The ions from the body fluid that participate in the corrosion layer formation were identified by analyzing layers formed in different solutions that contain only some of the ions of SBF. The layer composition was analyzed by different complementary methods. We used energy dispersive X-ray analysis, grazing incidence X-ray diffraction, and Fourier transform infrared spectroscopy. The morphology of the corrosion layers was studied using scanning electron microscopy and light microscopy. In m-SBF with and without albumin we found an amorphous layer of carbonated calcium phosphate with some calcium replaced by magnesium. It can be clearly shown that calcium is only deposited in the corrosion layer if phosphates are in the solution. The cross-sections reveal that there are some sharp crevices in the substrate. The work systematically explores the nature of surface layers on magnesium rare-earth alloys formed in complex SBFs, with the aim to elucidate the influence of specific electrolyte components on the morphology, structure, and composition of corrosion layers on Mg alloys. PMID- 18286624 TI - Platelet and endothelial adhesion on fluorosurfactant polymers designed for vascular graft modification. AB - A prominent failure mechanism of small diameter expanded polytetrafluoroethylene (ePTFE) vascular grafts is platelet-mediated thrombosis. We have designed a surface modification for ePTFE consisting of a self-assembling fluorosurfactant polymer (FSP) bearing biologically active ligands, including adhesive peptides and polysaccharide moieties. The goal of this biomimetic construct is to improve graft hemocompatibility by promoting rapid surface endothelialization, whereas minimizing platelet adhesion. Here we present a direct comparison of platelet and endothelial cell (EC) adhesion to FSPs containing one of three cell-adhesion peptides: cyclic Arg-Gly-Asp-D-Phe-Glu (cRGD), cyclic *Cys-Arg-Arg-Glu-Thr-Ala Trp-Ala-Cys* (cRRE, *denotes disulfide bond cyclization), linear Gly-Arg-Gly-Asp Ser-Pro-Ala (RGD), or a polysaccharide moiety: oligomaltose (M-7), later designed to prevent nonspecific protein adhesion. Measurements of soluble peptide-integrin binding indicated that cRRE exhibits very low affinity for the alpha(IIb)beta(3) platelet fibrinogen receptor. Static and dynamic adhesion of washed, activated platelets on FSP-modified surfaces revealed that M-7 and cRRE promote significantly less platelet adhesion compared to RGD and cRGD FSPs, whereas EC adhesion was similar on all peptide FSPs and minimal on M-7 FSP. These results illustrate the potential for ligands presented in a FSP surface modification to selectively adhere ECs with limited platelet attachment. PMID- 18286625 TI - Assembly of multilayer PSS/PAH membrane on coherent alginate/PLO microcapsule for long-term graft transplantation. AB - Conventional alginate/poly-L-ornithine (AP) membranes used to immunoisolate foreign tissue transplants fail in long-term transplantations of immortal cell lines. We have developed a novel layer-by-layer (LbL) membrane using polystyrene sulfonate and polyallylamine hydrochloride (PSS/PAH) on top of the coherent AP membrane. Assembly of the LbL membrane was followed by electrophoresis, and the surface morphologies and structure were characterized and examined by cryo scanning electron microscope and transmission electron microscopy. Unlike the standard AP membrane, the LbL membrane withstood the internal pressure generated by continuous cell proliferation of microencapsulated HEK-293 and Min-6 cells. The new membrane did not affect insulin secretion or diffusion by Min-6 cells. PMID- 18286626 TI - Effects of reduction on the denaturation kinetics of human hair. AB - Although human hair as an alpha-keratinous fiber exhibits a complex morphology, it can be considered as a nano-structured filament/matrix composite for the context of thermal analysis. Using differential scanning calorimetry (DSC) in water, the denaturation performance of the alpha-helical protein fraction and the effects of reductive treatments were studied. The results are viewed in the context of a previous study for oxidative treatments. It was found that the course of the denaturation process remains generally unperturbed by the treatment, following an irreversible, one-step, first-order process. Arrhenius activation energies and pre-exponential factors were determined from the DSC curves by applying the principles of the Friedman-method. Comparing activation energy values between reductive and oxidative processes shows the differences of the effects on the components of the composite. In contrast, the values of the rate constant at the denaturation temperature, though showing differences in their trends with cumulative treatments, are very similar. This further emphasizes the theory that the viscosity of the matrix affects strict kinetic control over the denaturation of the alpha-helical segments. Once the viscosity of the matrix has decreased enough for the denaturation process to occur, this follows a path that is largely independent of the temperature range and of the chemical pre-history. PMID- 18286627 TI - A new view of protein synthesis: mapping the free energy landscape of the ribosome using single-molecule FRET. AB - This article reviews the application of single-molecule fluorescence resonance energy transfer (smFRET) methods to the study of protein synthesis catalyzed by the ribosome. smFRET is a powerful new technique that can be used to investigate dynamic processes within enzymes spanning many orders of magnitude. The application of wide-field smFRET imaging methods to the study of dynamic processes in the ribosome offers a new perspective on the mechanism of protein synthesis. Using this technique, the structural and kinetic parameters of tRNA motions within wild-type and specifically mutated ribosome complexes have been obtained that provide valuable new insights into the mechanism and regulation of translation elongation. The results of these studies are discussed in the context of current knowledge of the ribosome mechanism from both structural and biophysical perspectives. PMID- 18286628 TI - Infrared spectroscopic study of the binding of divalent cations to Akazara scallop troponin C: the effect of the methylene side chain of glutamate residue. AB - Akazara scallop striated adductor muscle troponin C (TnC) binds only one Ca2+ because the three EF-hand motifs are short of critical residues for the coordination of Ca2+. Fourier-transform infrared spectroscopy was applied to study coordination structures of M2+ (= Mg2+, Ca2+, Sr2+, and Ba2+) bound in an Akazara scallop TnC mutant (E142D) and the wild-type TnC C-lobe in D2O solution. The region of the COO- antisymmetric stretch provides information regarding the coordination modes of a COO- group to a metal ion. The side chain COO- group of Asp142 did not bind to Ca2+ in the bidentate coordination mode, suggesting that the absence of a methylene group is critical for the Ca2+ coordination structure of Akazara scallop TnC (Nara et al., Vib Spect 2006, 42, 188-191). The present study has shown that the absence of a methylene group is not compensated for by a larger metal ion such as Sr2+ or Ba2+. CD spectra showed that the secondary structures are conserved between M2+-free (apo), Mg2+-loaded, Ca2+-loaded, Sr2+ loaded, and Ba2+-loaded states, which was consistent with the results estimated from their amide I band patterns. The metal-ligand interaction at position 12 of site IV is discussed in comparison with the coordination mode of the side chain COO- group of the wild-type TnC C-lobe. PMID- 18286629 TI - In vitro interaction between calf thymus DNA and Escherichia coli LPS in the presence of divalent cation Ca2+. AB - With increasing addition of Escherichia coli LPS to calf thymus DNA, both dissolved in CaCl2, absorption maxima of DNA at 260 nm decreased gradually with the appearance of isosbastic points at both ends of spectra, which implied some binding between DNA and LPS. Hill plot of absorbance data showed that the binding interaction was positive cooperative in nature. For any fixed concentration of DNA and LPS, extent of interaction increased as concentration of CaCl2 was raised from 1.0 to 100 mM, signifying the electrostatic nature of the interaction, mediated through Ca2+ ion. Stepwise addition of EDTA, a chelating agent for divalent cations, to DNA-LPS bound complex gradually reversed the spectral shift with increase in absorbance at 260 nm, which implied opening up of the complex, that is, reversible nature of the interaction. Circular dichroism spectral changes of DNA by the addition of LPS indicated partial transition of DNA from B to A form. Isothermal titration calorimetric (ITC) study showed that the DNA-LPS binding was an exothermic and enthalpy-driven phenomenon. Moreover, in the presence of 100 mM CaCl2, binding constant of the interaction was found to be 2.6 x 10(4) M(-1) and 3.1 x 10(4) M(-1) from the analysis of Hill plot and ITC result, respectively. DNA-melting study showed that the LPS binding had increased the melting temperature of DNA, indicating more stabilization of DNA double helix. The binding of LPS to DNA made the complex resistant to digestion with endonucleases EcoRI and DNase I. PMID- 18286630 TI - Comparative cellular morphology suggesting the existence of resident dendritic cells within immune organs of salmonids. AB - This report is the first morphological description of cells that resemble dendritic cells, which appear to form resident populations within the spleen and anterior kidney of fish. Based on examination of three salmonid species, including, rainbow trout, brook trout, and Atlantic salmon, the cells were most abundant in the spleen, although they were always present in the anterior kidney. The cells appeared diffusely distributed, often near blood vessels of the spleen and kidney of healthy fish and within the epithelium, connective tissue, and blood vessels of rainbow trout gills with experimentally induced microsoporidial gill disease. The dendritic-like cells in this study contained granules that resemble Birbeck granules, which are considered to be morphological markers of Langerhans cells in mammals. The cells were approximately 6 mum in diameter and contained Birbeck-like (BL) granules localized near centrioles. Although the dendritic-like cells in the three salmonid species shared many similarities, morphological differences were found in the fine structure of the rod portion of the BL granules. Rainbow trout BL granules contained amorphous material, while the other salmonid species contained particulate material arranged in a square lattice arrangement. The BL granules in the cells of Atlantic salmon had a narrow diameter and contained four layers of particulate material when sectioned longitudinally; two layers enveloped by the granule membrane and two central layers making up a central lamella, which is common in mammalian Birbeck granules. PMID- 18286632 TI - Association study of brain-derived neurotrophic factor (BDNF) and LIN-7 homolog (LIN-7) genes with adult attention-deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) is a common psychiatric disorder with a large genetic component that has been shown to persist into adulthood in 30-60% of childhood ADHD cases. Adult ADHD confers an increased risk of ADHD in relatives when compared to childhood ADHD, possibly due to a greater genetic liability than the childhood form. Brain-derived neurotrophic factor (BDNF) is a neurotrophin expressed in the brain throughout life and is involved in survival, differentiation, and synaptic plasticity of several neuronal systems including dopaminergic pathways. Mammalian LIN-7 homolog is selectively expressed in specific neuronal populations and is involved in the postsynaptic density of neuronal synapses. LIN-7 is also a positional candidate, as it lies immediately downstream of BDNF. We tested for association between five BDNF polymorphisms, two LIN-7 polymorphisms and adult ADHD. The sample consisted of 80 trios comprised of an adult ADHD proband and their biological parents and an independent sample of 121 adult ADHD cases and a corresponding number of sex, age, and ethnically matched controls (total 201 probands). Allelic and haplotype association was found between both BDNF and adult ADHD, and LIN-7 and adult ADHD. HapMap indicates BDNF and LIN-7 occur in different haplotype blocks, though some linkage disequilibrium exists between the SNPs in these adjacent genes. Further investigations into the pathologic mechanisms of BDNF and LIN-7 in adult ADHD are required. PMID- 18286631 TI - Externalizing disorders in American Indians: comorbidity and a genome wide linkage analysis. AB - Alcohol dependence is one of the leading causes of morbidity and mortality in Native Americans. Externalizing disorders such as conduct disorder (CD) and antisocial personality disorder (ASPD) have been demonstrated to have significant comorbidity with alcohol dependence in the general population. This study's aims were to: assess the comorbidity of DSM-III-R ASPD and CD with alcohol dependence, to map susceptibility loci for ASPD and CD, and to see if there is overlap with loci previously mapped for alcohol dependence phenotypes in 587 American Indians. Alcohol dependence was found to be comorbid with DSM-III-R ASPD but not CD. However, the amount of alcohol dependence in the population attributable to ASPD and/or CD is low. ASPD and the combined phenotype of participants with ASPD or CD were both found to have significant heritability, whereas no significant evidence was found for CD alone. Genotypes were determined for a panel of 791 micro satellite polymorphisms in 251 of the participants. Analyses of multipoint variance component LOD scores, for ASPD and ASPD/CD, revealed six locations that had a LOD score of 2.0 or above: on chromosome 13 for ASPD and on chromosomes 1, 3, 4, 14, 17, and 20 for ASPD/CD. These results corroborate the importance of several chromosomal regions highlighted in prior segregation studies for externalizing diagnoses. These results also further identify new regions of the genome, that do not overlap with alcohol dependence phenotypes previously identified in this population, that may be unique to either the phenotypes evaluated or this population of American Indians. PMID- 18286633 TI - Autism and serotonin transporter gene polymorphisms: a systematic review and meta analysis. AB - The serotonin transporter gene (5-HTT) plays a crucial role in serotonergic neurotransmission and has been found to be associated, with varying degrees of significance, with many diseases, including autism. Prior association studies of autism have yielded conflicting results regarding the association between two common 5-HTT polymorphisms, the promoter insertion/deletion (5-HTTLPR) and the intron 2 VNTR (STin2 VNTR). We conducted a systematic review and meta-analysis to test the following hypotheses: (i) there is an association between autism and either or both of the 5-HTTLPR and STin2 VNTR polymorphisms, and (ii) the S allele of 5-HTTLPR and/or the STin2.12 allele of the VNTR are the specific risk alleles for autism. All published family-based and population based studies were examined to determine the overall strength of association between 5-HTT polymorphisms and autism. After exclusion of studies with overlapping samples and studies whose data did not allow for calculation of an odds ratio, 16 studies were included for final analyses, all but two of which used a family-based design. The meta-analysis failed to find a significant overall association between either of the 5-HTT polymorphisms examined and autism. Further, no allelic transmission distortion was found when studies of simplex (11 studies) and multiplex (3 studies) family samples were analyzed separately. However, there was significant heterogeneity by ethnicity; family based studies of US mixed population samples showed preferential transmission of the S allele of 5-HTTLPR (S allele:L allele = 247:183), while there was no allelic distortion among the family-based studies of European and Asian samples. PMID- 18286634 TI - Reduced hippocampal volumes associated with the long variant of the tri- and diallelic serotonin transporter polymorphism in major depression. AB - Substantial evidence supports a role for dysfunction of the serotonin transporter (5-HTT) in the pathogenesis of major depression. The polymorphism of the serotonin transporter gene (5-HTTLPR) was found to be associated with reduced hippocampal volume in major depression. However, the original diallelic polymorphism was criticized, because the L-allele can be subtyped into La and Lg alleles, the latter of which is thought to be similar to the S-allele. Therefore, the study aim was to examine the influences of the triallelic (La-Lg-S system) and diallelic 5-HTTLPR on hippocampal volumes in patients with major depression and healthy controls. Using high-resolution MRI hippocampal volumes and polymorphisms (5-HTTLPR) were measured in 60 in-patients with major depression and 60 healthy controls. Patients with the La/La genotype had significantly smaller hippocampal gray and white matter than La/La controls. No significant differences were found between patients and controls with La/(Lg + S) or (Lg + S)/(Lg + S) genotype. Moreover, within the patient group the La/La homozygous genotype had significantly smaller hippocampal white matter volumes than the La/(Lg + S) or (Lg + S)/(Lg + S) genotype. In conclusion, with the diallelic as well as the triallelic system the homozygosity for the long-allele is associated with decreased hippocampal volumes in patients with major depression, but not in healthy controls, suggesting that disease or stress specific processes linked to the serotonergic system may enhance the vulnerability to morphological alterations. PMID- 18286635 TI - Symptoms and physical activity behavior in individuals with multiple sclerosis. AB - We examined overall and specific symptoms as correlates of physical activity in individuals with multiple sclerosis (MS). Participants (N = 133) completed questionnaires that measured overall symptoms; and specific symptoms of depression, pain, and fatigue; difficulty walking; and physical activity. Initial analyses indicated that higher levels of overall symptoms (r = -.50), fatigue (r = -.26), and difficulty walking (r = -.46) were associated with lower levels of physical activity. Path analysis demonstrated that higher levels of overall symptoms were directly and indirectly associated with lower levels of physical activity; the indirect pathway involved difficulty walking (gamma beta = -.17). Such findings indicate that walking difficulty may partially explain the negative relationship between overall symptoms and physical activity behavior in MS. PMID- 18286636 TI - Molecularly designed surfaces for blood deheparinization using an immobilized heparin-binding peptide. AB - Systemic heparinization, used during haemodialysis to prevent blood clotting on the extracorporeal circuit, leads to a high incidence of hemorrhagic complications. The adverse reactions associated with heparin neutralization using protamine sulphate justify the development of an alternative system for blood deheparinization. The main objective of this work is to design nanostructured surfaces with the capacity to bind heparin from blood in a selective way. A heparin-binding polypeptide, composed of L-lysine and L-leucine (pKL), was synthesized and immobilized, in different concentrations, onto self-assembled monolayers (SAMs) terminated with tetra(ethylene-glycol) (EG4 SAMs). Immobilization was performed using a fixed concentration of pKL after surface activation to different degrees using a range of CDI (N,N'-carbonyldiimidazole) concentrations. Results demonstrated that the presence of pKL increases heparin adsorption to EG4-SAMs, independently of the pKL concentration and the way of immobilization (adsorption or covalent bound). Selectivity towards heparin was successfully achieved on SAMs with low concentrations of immobilized pKL (9-17% of pKL). Surfaces were characterized using ellipsometry, contact angle measurements, Fourier transform infrared reflection absorption spectroscopy (IRAS), atomic force microscopy, and X-ray photoelectron spectroscopy. Heparin adsorption was assessed using IRAS and N-sulphonate-(35)S-heparin. Therefore, this study could give a good contribution for the design of blood deheparinization devices. PMID- 18286637 TI - Activation of human leukocytes on tantalum trabecular metal in comparison to commonly used orthopedic metal implant materials. AB - We analyzed leukocyte functions and cytokine response of human leukocytes toward porous tantalum foam biomaterial (Trabecular Metaltrade mark, TM) in comparison to equally sized solid orthopedic metal implant materials (pure titanium, titanium alloy, stainless steel, pure tantalum, and tantalum coated stainless steel). Isolated peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophil leukocytes (PMN) were cocultured with equally sized metallic test discs for 24 h. Supernatants were analyzed for cytokine content by enzyme-linked immunosorbent assay. Compared to the other used test materials there was a significant increase in the release of IL (interleukin)-1ra and IL-8 from PMN, and of IL-1ra, IL-6, and TNF-alpha from PBMC in response to the TM material. The cytokine release correlated with surface roughness of the materials. In contrast, the release of IL-2 was not induced showing that mainly myeloid leukocytes were activated. In addition, supernatants of these leukocyte/material interaction (conditioned media, CM) were subjected to whole blood cell function assays (phagocytosis, chemotaxis, bacterial killing). There was a significant increase in the phagocytotic capacity of leukocytes in the presence of TM-conditioned media. The chemotactic response of leukocytes toward TM-conditioned media was significantly higher compared to CM obtained from other test materials. Furthermore, the bactericidal capacity of whole blood was enhanced in the presence of TM-conditioned media. These results indicate that leukocyte activation at the surface of TM material induces a microenvironment, which may enhance local host defense mechanisms. PMID- 18286638 TI - Synthesis, self-assembly in water, and cytotoxicity of MPEG-block-PLLA/DX conjugates. AB - Docetaxel (DX) is one of the most effective antineoplastic drugs. Its current clinical administration is limited because of its hydrophobicity and serious side effects. A polymer/DX conjugate is designed and successfully prepared to solve these problems. It is monomethoxy-poly(ethylene glycol)-block-poly(L-lactide)/DX (MPEG-PLLA/DX). It was synthesized by reacting DX with carboxyl-terminated copolymer MPEG-PLLA, which was prepared by reacting succinic anhydride with hydroxyl-terminated copolymer monomethoxy-poly(ethylene glycol)-block-poly(L lactide) (MPEG-PLLA). Its structure and molecular weight was confirmed by (1)H NMR and GPC. The MPEG-PLLA/DX micelles in aqueous solution were prepared using a solvent displacement method and characterized by dynamic light scattering for size and size distribution, and by transmission electron microscopy for surface morphology. Its antitumor activity against HeLa cancer cells evaluated by MTT assay showed that it had a similar antitumor activity to pure DX at the same drug content. PMID- 18286639 TI - Functional and phenotypic characterization of human keratinocytes expanded in microcarrier culture. AB - Skin cells for transplantation are routinely prepared by growing patient keratinocytes in a semi-defined cocktail of growth factors, including serum and feeder cells. However, these reagents require substantial risk remediation and can contribute to transplant rejection. Microcarrier culture is an emerging technology that may allow the elimination of feeder cells whilst facilitating expansion of cultured keratinocytes. However, the behavior of keratinocytes in microcarrier culture and the potential of these cells to form an epidermis have been poorly defined. We characterized freshly isolated human keratinocytes cultured on CultiSpher-G microcarriers in the absence of murine feeder cells and assessed the potential of the keratinocytes to form an epidermis in an in vitro model. In a single passage, keratinocytes multiplied 44.9-fold in microcarrier bioreactor culture in 17 days, whereas two-dimensional cultures reached confluence in 9 days and only expanded 7.4-fold. Histological characterization of keratinocytes on the microcarriers revealed that the cells were randomly distributed within these porous structures, however, not all pores contained cells. High-resolution microcomputed tomography imaging of the microcarriers confirmed limited interconnectivity of the pores. Immunoreactivity of specific epidermal markers was confirmed during cell expansion via immunohistochemistry. Despite the expression of differentiation markers, microcarrier-expanded keratinocytes retained the capacity to form an epidermis, as was evaluated using an in vitro human skin equivalent model. The epidermis formed by microcarrier expanded keratinocytes in this model exhibited morphology similar to native skin. Significantly, the microcarrier technique successfully eliminates the need for a feeder cell layer and hence facilitates development of an improved culture system. PMID- 18286640 TI - New class of anti-microbial agents: synthesis, characterization, and anti microbial activities of metal chelated polyurea. AB - A new class of metal chelated polyurea have been synthesized by the reaction of toluene 2,4-diisocyanate (TDI) with chelated Schiff base diamines. The synthesized polyurea have been characterized by analytical, spectral, and thermal analysis. The results of TGA ascribed that [Cu(II)-PoU(A)] show better heat resistant properties than other metals chelated polyurea. The antibacterial activities of all the synthesized polymers were determined using the shaking flask method, where 30 mg/mL concentrations of each compound were tested against 10(5) CFU/mL solutions of S. aureus, E. coli, B. subtillis, S. typhi. The number of viable bacteria was calculated by using the spread plate method on agar plates and the number of viable bacteria was counted after 24 h of incubation period at 37 degrees C. All the polymers showed good antibacterial activity. The Cu(II) chelated polyurea show higher zone of inhibition then other due to higher stability constant and may be used in biomedical applications. PMID- 18286641 TI - Analysis of the osteoinductive capacity and angiogenicity of an in vitro generated extracellular matrix. AB - In this study, the osteoinductive potential of an in vitro generated extracellular matrix (ECM) deposited by marrow stromal cells seeded onto titanium fiber mesh scaffolds and cultured in a flow perfusion bioreactor was investigated. Culture periods of 8, 12, and 16 days were selected to allow for different amounts of ECM deposition by the cells as well as ECM with varying degrees of maturity (Ti/ECM/d8, Ti/ECM/d12, and Ti/ECM/d16, respectively). These ECM-containing constructs were implanted intramuscularly in a rat animal model. After 56 days, histologic analysis of retrieved constructs revealed no bone formation in any of the implants. Surrounding many of the implants was a fibrous capsule, which was often interspersed with fat cells. Within the pore spaces, the predominant tissue response was the presence of blood vessels and young fibroblasts or fat cells. The number of blood vessels on a per area basis calculated from a histomorphometric analysis increased as a function of the amount of ECM within the implanted constructs, with a significant difference between Ti/ECM/d16 and plain Ti constructs. These results indicate that although an in vitro generated ECM alone may not induce bone formation at an ectopic site, its use may enhance the vascularization of implanted constructs. PMID- 18286642 TI - Decrease in factor V activity: a new adverse effect of purine analogs in inflammatory bowel diseases? PMID- 18286643 TI - Identification of a molecular target for glutamate regulation of astrocyte water permeability. AB - Astrocytes play a key role for maintenance of brain water homeostasis, but little is known about mechanisms of short-term regulation of astrocyte water permeability. Here, we report that glutamate increases astrocyte water permeability and that the molecular target for this effect is the aquaporin-4 (AQP4) serine 111 residue, which is in a strategic position for control of the water channel gating. The glutamate effect involves activation of group I metabotropic glutamate receptors (mGluR), intracellular calcium release, and activation of calcium/calmodulin-dependent protein kinase II (CaMKII) and nitric oxide synthase (NOS). The physiological impact of our results is underlined by the finding that mGluR activation increases the rate of hypoosmotic tissue swelling in acute rat hippocampal slices. Cerebral ischemia is associated with an excessive release of glutamate, and in postischemic cerebral edema ablation of AQP4 attenuates the degree of damage. Thus, we have identified AQP4 as the molecular target for drugs that may attenuate the development of brain edema. PMID- 18286644 TI - Qualitative investigation of patient adherence to 5-aminosalicylic acid therapy in patients with ulcerative colitis. AB - BACKGROUND: There has been a lack of qualitative research specifically in patients with inflammatory bowel disease relating to reasons for failure to take medication. We aimed to address this gap and also identify factors which might increase adherence. METHODS: Twenty-seven patients from 3 UK hospital sites (Leicester, Norwich, and Cardiff) were recruited to take part in a qualitative study based on semistructured interviews. RESULTS: A model was developed to illustrate the way in which patients appear to balance the benefits and disadvantages of taking 5-aminosalicylic acid (5-ASA) medication. The degree of information held by patients regarding ulcerative colitis (UC) and self evaluation of the benefits of 5-ASA appears to impact whether patients accept or reject the medication. Decision-making on an ill-informed basis may be a factor leading to a reduction in adherence. Due to the nature of the condition, adherence levels may be affected by potential difficulties in terms of experiencing and understanding the links between the medication and health outcomes. CONCLUSIONS: Important determinants of adherence to 5-ASA medication in patients with UC appear to include the level of information provided and patient beliefs about prescribed 5-ASA. Patient adherence to 5-ASA requires encouragement and reinforcement and the patient-clinician relationship has a crucial role to play in this dynamic. PMID- 18286645 TI - Kir4.1 and AQP4 associate with Dp71- and utrophin-DAPs complexes in specific and defined microdomains of Muller retinal glial cell membrane. AB - The dystrophin-associated proteins (DAPs) complex consisting of dystroglycan, syntrophin, dystrobrevin, and sarcoglycans in muscle cells is associated either with dystrophin or its homolog utrophin. In rat retina, a similar complex was found associated with dystrophin-Dp71 that serves as an anchor for the inwardly rectifying potassium channel Kir4.1 and the aqueous pore, aquaporin-4 (AQP4). Here, using immunofluorescence imaging of isolated retinal Muller glial cells and co-immunoprecipitation experiments performed on an enriched Muller glial cells end-feet fraction, we investigated the effect of Dp71 deletion on the composition, anchoring, and membrane localization of the DAPs-Kir4.1 and/or -AQP4 complex. Two distinct complexes were identified in the end-feet fraction associated either with Dp71 or with utrophin. Upon Dp71 deletion, the corresponding DAPs complex was disrupted and a compensating utrophin upregulation was observed, accompanied by diffuse overall staining of Kir4.1 along the Muller glial cells and redistribution of the K(+) conductance. Dp71 deficiency was also associated with a marked reduction of AQP4 and beta-dystroglycan expression. Furthermore, it was observed that the Dp71-DAPs dependent complex could be, at least partially, associated with a specific membrane fraction. These results demonstrate that Dp71 has a central role in the molecular scaffold responsible for anchoring AQP4 and Kir4.1 in Muller cell end-feet membranes. They also show that despite its close relationship to the dystrophin proteins and its correlated upregulation, utrophin is only partially compensating for the absence of Dp71 in Muller glial cells. PMID- 18286646 TI - Urinary leukotriene E4 excretion: a biomarker of inflammatory bowel disease activity. AB - BACKGROUND: Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory disorders collectively referred to as inflammatory bowel diseases (IBD). Cysteinyl leukotrienes are proinflammatory 5-lipoxygenase-derived products that play a major role in the immune and inflammatory response. Consequently, they may be involved in the pathogenesis of IBD. The aim of this study was therefore to evaluate 1) the urinary excretion of leukotriene E(4) (LTE(4)) in IBD patients and healthy volunteers, and 2) the association between LTE(4) production and the activity (relapse/remission) of the disease. METHODS: IBD patients and healthy volunteers were prospectively recruited. CD and UC activity was determined on inclusion with the Crohn's Disease Activity Index and Clinical Activity Index, respectively. Urine was collected and the urinary excretion of LTE(4) was measured by liquid chromatography tandem mass spectrometry. RESULTS: 32 CD patients, 28 UC patients, and 30 controls were enrolled in the study. LTE(4) urinary excretion was significantly increased (P < 0.01) in CD [52.0 pg/mg creatinine (10th-90th percentiles: 26.2-148.0)] and UC [64.1 pg/mg creatinine (10th-90th percentiles: 26.7-178.0)] patients compared to controls [32.3 pg/mg creatinine (10th-90th percentiles: 21.8-58.8)]. LTE(4) levels were higher (P < 0.001) in patients with active disease than in patients in remission, for whom the levels of LTE(4) were similar to the levels of controls. CONCLUSIONS: Cysteinyl leukotriene pathway activation could contribute to the inflammation associated with IBD. The quantification of urinary LTE(4) could be an interesting noninvasive biomarker for the assessment of IBD activity. PMID- 18286647 TI - Current understanding of fungal microflora in inflammatory bowel disease pathogenesis. AB - Inflammatory bowel diseases are a current and growing public health problem, with a prevalence that appears to be increasing in most countries and cultures. While most research into the triggering phenomenon has focused on the interaction between commensal bacteria and inflammatory bowel disease, enteric fungi may also be important in determining disease susceptibility. Herein we review what is known about enteric fungi and the mechanisms by which they and their dysregulation might be involved in triggering inflammatory diseases of the bowel. PMID- 18286649 TI - Penile replantation, complication management, and technique refinement. AB - We describe a case of complete guillotine-type penile amputation at the proximal penile shaft. The blood flow was established 10 h after trauma. Circulation in the replanted penis was quite good but there was progressive prepuce necrosis after the hematoma. Cosmetic and urinary outcome was good 6 weeks later. The repair of deep dorsal penile vessels helps in corpus tissue healing and glans circulation. The blood supply from the corpus tissue is sufficient for the survival of the replanted penis even when the repaired dorsal vessels were occluded. Surgical pitfalls in replantation procedures and complication management are discussed. PMID- 18286648 TI - The ABCA1 cholesterol transporter associates with one of two distinct dystrophin based scaffolds in Schwann cells. AB - Cytoskeletal scaffolding complexes help organize specialized membrane domains with unique functions on the surface of cells. In this study, we define the scaffolding potential of the Schwann cell dystrophin glycoprotein complex (DGC) by establishing the presence of four syntrophin isoforms, (alpha1, beta1, beta2, and gamma2), and one dystrobrevin isoform, (alpha-dystrobrevin-1), in the abaxonal membrane. Furthermore, we demonstrate the existence of two separate DGCs in Schwann cells that divide the abaxonal membrane into spatially distinct domains, the DRP2/periaxin rich plaques and the Cajal bands that contain Dp116, utrophin, alpha-dystrobrevin-1 and four syntrophin isoforms. Finally, we show that the two different DGCs can scaffold unique accessory molecules in distinct areas of the Schwann cell membrane. Specifically, the cholesterol transporter ABCA1, associates with the Dp116/syntrophin complex in Cajal bands and is excluded from the DRP2/periaxin rich plaques. PMID- 18286650 TI - Comment on: Microsurgical arterovenous loops and biological templates: a novel in vivo chamber for tissue engineering. PMID- 18286651 TI - A new training method to improve deep microsurgical skills using a mannequin head. AB - Neurosurgeons need fine and special microsurgical techniques, such as the ability to suture deep microvasculature. Intensive training is required to perform microsurgery, especially in deep microvascular anastomosis. There have been many previous reports of training methods for typical microsurgical techniques, including suturing of surgical gloves, Silastic tubes, living animals, and chicken wing arteries. However, there have been no reports of training methods to improve deep microsurgical skills under the various hand positions specific to neurosurgical operation. Here, we report a new training method using a mannequin head, water balloons, and clay to mimic actual deep microsurgery in the brain. This method allows trainees to experience microsurgery under various hand positions to approach the affected areas located at various depths in the brain from various angles. PMID- 18286652 TI - Hepatic lobectomy and segmentectomy models using microsurgical techniques. AB - The mouse and rat models are the most commonly utilized experimental models of partial liver resection. Microscope-assisted partial hepatectomy is associated with better outcomes by reducing the risk of vena cava stenosis after ligature and injury to other liver pedicles. In addition, microsurgery allows lobectomies and segmentectomies after individual ligation of intrahepatic vascular branches, including the left and right segments of the median lobe, which has not been reported before. Here, we describe a new technique (catheter-assisted suture technique) and report our experience with partial hepatectomies in rodents using a microsurgical approach. PMID- 18286653 TI - Endogenous adipose tissue as a hemostatic: use in microsurgery. AB - Bleeding is a frequent complication of microsurgical repair of small blood vessels and time is spent while hemostasis is accomplished. We studied the hemostatic effect of endogenous adipose tissue on bleeding from rat femoral arterial anastomoses. We measured bleeding time (time from removal of clamps to cessation of active bleeding) and mean arterial blood velocity (using a micro Doppler system), the latter immediately after anastomosis, and again 7 days post anastomosis. Bleeding time for vessels with fat applied to the artery was 50% less than when no fat was applied. Blood velocity by day 7 post-anastomosis returned to values equivalent to those for intact arteries. Histological evaluation of the anastomotic site demonstrated no significant differences in inflammatory response between fat-treated and untreated arteries. These data suggest that endogenous adipose tissue may be a useful hemostatic agent devoid of significant effects on small artery blood velocity or histology. PMID- 18286654 TI - Improved long-term recording of nerve signal by modified intrafascicular electrodes in rabbits. AB - Methods for long-term recording of peripheral nerve activity via intrafascicular electrodes have not been optimized. We compared the long-term functionality of custom-made 95%Pt/5%Ir intrafascicular electrodes containing a proximal spring like structure to that of conventional straight electrodes. The modified electrode was implanted into the sciatic nerve fascicle of a random hind limb in 14 rabbits for 9 months. A conventional electrode was implanted in the opposite hind limb as a control. Orthodromic and antidromic nerve potentials were sampled and analyzed monthly. Latency, amplitude, and nerve conduction velocity of electrical signals were generally similar within the modified group and straight control group at different time intervals (P > 0.05). However, at the conclusion of the study period, the modified electrode group had a greater number of functioning electrodes (P < 0.05) and a greater total functioning electrode time (P = 0.006). Intrafascicular electrodes with a spring-like structure demonstrated superior potential for long-term electrophysiological monitoring over straight electrodes. PMID- 18286655 TI - Free flap coverage of gluteal defect using posteriorly translocated deep femoral vessels as recipient. AB - A 63-year-old paraparetic man presented with a large ischial pressure sore over the left buttock region. A failed surgical attempt using gluteal thigh flap transfer rendered the reconstructive task a challenging issue. In the absence of suitable regional flaps or any accessible recipient vessels nearby, we must employ a novel strategy to solve the clinical problem. The entire length of the deep femoral vessel was isolated through the anterior approach, which was then transposed posterior wards to the gluteal region to nourish a latissimus dorsi myocutaneous free flap while the patient was laid prone. The wound healed uneventfully and the patient remains ulcer-free throughout 3 years of follow-up. PMID- 18286656 TI - The epineural sleeve technique for nerve graft reconstruction enhances nerve recovery. AB - The purpose of the study was evaluation of nerve recovery following epineural sleeve technique for graft reconstruction in rat sciatic nerve. This technique provides the epineural sleeve to cover and separate the site of coaptation. Animals were divided into three groups: CNG-conventional nerve grafting, ESN epineural sleeve from recipient nerve stumps, ESG-epineural sleeve from graft. Nerve regeneration was evaluated by pin-prick, toe-spread test, walking track analysis and somatosensory-evoked potentials (SEP), gastrocnemius index (GI), and histomorphometric evaluation. Most parameters (SFI, SEPs, and GI) showed significantly better nerve recovery for ESN group when compared to conventional CNG group. Also ESG group revealed better result for SFI. Better functional results for ESN and ESG groups were further confirmed by histomorphometric analysis: higher axon density and diameters as well as thicker myelin sheath. Epineural sleeve graft technique may be promising method with potential application for nerve reconstructive procedures. Better functional nerve recovery can be anticipated. PMID- 18286657 TI - Does the nicotine patch help in the postoperative period for arterial vessel repair? AB - This study attempts to determine whether a nicotine patch will affect the patency rate of the femoral artery anastomosis in a smoking rat model. Twenty-four rats underwent a smoking protocol of 4 weeks of exposure to cigarette smoke in a smoking chamber at 1 hour per day, using a 1:10 dilution of unfiltered cigarette smoke. Each rat then underwent a femoral artery transaction and anastomosis. Postoperatively, the rats were divided into two groups of 12. Group I had no postoperative smoking or nicotine therapy. Group II had nicotine patch treatment only. The femoral artery was evaluated for flow 7-days postoperatively. At the time of anastomosis, the femoral arteries were smaller in diameter, averaging 0.5 mm (0.4-0.7 mm) as opposed to the normal 0.75-1 mm. All anastomoses were patent at 5 min when the wound was closed. No difference in patency rate occurred with the use of the nicotine patch. PMID- 18286658 TI - Recording and stimulating properties of chronically implanted longitudinal intrafascicular electrodes in peripheral fascicles in an animal model. AB - The purpose of this experiment was to study the recording and stimulating properties, and biocompatibility of longitudinally implanted intrafascicular electrodes (LIFEs) in a rabbit sciatic nerve model when they were chronically implanted into peripheral fascicles. LIFEs were implanted chronically into sciatic nerve fascicles of rabbits as recording and stimulating electrodes. Motor evoked potentials (MEPs) and cortical somatosensory-evoked potentials (CSEPs) were recorded by using a transcranial stimulation system (TCS) over 6-month period to observe the change of the signals recorded. At the end of the experiment, the fascicles at the electrodes implanted site were anatomized for histological examination under light microscope and transmission electron microscope. Results showed onset latency (OL) of MEPs and CSEPs had no obvious change during the first month. However, OL significantly increased during the second month, and then became stable 3 months after implantation. The interpeak amplitudes (IPAs) of MEPs had no distinct change during the first month, but significantly decreased over the next period, and then became stable 3 months after implantation. The IPAs of CSEPs, however, decreased slowly over the 6-month period of the study. At the end of the experiment, histological examination indicated that a typical foreign body reaction developed, and electrodes caused mild damage to the fascicles, though inflammatory cells and neuroma were not seen around the electrodes. In conclusion, LIFEs have excellent recording and stimulating characters in addition to biocompatibility with peripheral fascicles. They can be implanted chronically into fascicles and record signals. PMID- 18286659 TI - Incidence and time of intraoperative vascular complications in head and neck microsurgery. AB - Vascular occlusion is still the main reason for flap loss and occurs mostly within the first hours after performing anastomoses. Many surgeons still prefer to perform reconstruction and close the defect before starting to anastomose. The aim of this investigation was to find out if detection of early vascular occlusion is facilitated with a prolonged observation period. Between January 2000 and August of 2006, 350 consecutive free flap transfers for reconstruction in maxillofacial surgery were analyzed. In all flaps vascular anastomoses were performed prior to definite flap insertion. The flaps were controlled continuously during soft tissue or bony reconstruction until final wound closure at the neck. Complete operation time, ischemia time of the flap, and time from reperfusion to wound closure (direct pedicle observation time) were registered for each flap. In 350 flaps (138 radial forearm, 94 fibular, 53 ALT, 23 DCIA, 26 soleus perforator, 9 lateral arm, 5 lat. dorsi, and 2 scapular), operation time in average was 8.5 h, ischemia time varied between 78 and 139 min (average 104 min), and direct pedicle observation time of the flaps was 144 min in average (93 192 min). Four arteries and 2 veins showed immediate failure within 5 min after clamp removal, 10 arteries and 6 veins developed thromboses during the direct pedicle observation time. Out of these 16 vascular complications, 15 developed later than 15 min, 7 of them later than 30 min, and 2 of them later than 45 min. The overall complication rate including secondary revision of the pedicle was 16.8%, and an overall flap survival rate resulted in 95.4%. We conclude that microvascular anastomoses should be controlled for at least 45 min before definite wound closure. By performing anastomoses first and flap insertion second, this can be easily warranted. PMID- 18286661 TI - Fragmentation patterns of new esterified and unesterified aromatic 1 hydroxymethylene-1, 1-bisphosphonic acids by ESI-MSn. AB - 1-Hydroxymethylene-1,1-bisphosphonic acids (HMBPs) are compounds that have interesting pharmacological applications. Unfortunately few studies exist on their analyses by mass spectrometry (MS). In this work, we have analyzed new aromatic HMBPs and their prodrugs with electrospray tandem mass spectrometry (ESI MS(n)). We describe, for the first time, a complete study of fragmentation patterns, in both positive and negative-ion modes. In positive mode, the cation dissociations are mainly elimination of water and phosphorus fragments. In negative mode, losses of ROH (R==H, C(6)H(5), CH(3)OC(6)H(5)) and HPO(2) were observed. The results have revealed specific structural fingerprints for the screening of these compounds in complex biological mixtures. PMID- 18286660 TI - Establishment of duodenojejunal bypass surgery in mice: a model designed for diabetic research. AB - We have developed a mouse duodenojejunal bypass (DJB) surgical model that is for studying the effects of bariatric surgery on glucose homeostasis and has potential to impact clinical therapy of diabetes. The operation consists of using the majority of the duodenum and proximal part of the jejunum for biliopancreatic diversion. The distal end of the jejunum is anastomosed in an end-to-end fashion to the remaining proximal end of the duodenum just distal to the pylorus. The biliopancreatic secretions are diverted into the distal jejunum through an end-to side anastomosis. We performed 10 DJB operations in C57BL/6 mice, with a 100% survival rate. The surgery had no effect on the growth or feeding patterns of the animals. The intestinal mucosa showed normal histology and function. This study confirms that it is technically possible to perform DJB surgery in mice. This mouse model can be used in the study of surgical treatment for type II diabetes. PMID- 18286662 TI - Probing peptide libraries from Conus achatinus using mass spectrometry and cDNA sequencing: identification of delta and omega-conotoxins. AB - The peptide library present in the venom of the piscivorous marine snail Conus achatinus has been probed using a combination of mass spectrometry and cDNA sequencing methods. Matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) analysis, before and following global reduction/alkylation of peptide mixtures, permits the rapid classification of individual components on the basis of the number of disulfide bonds. Mass fingerprinting and the reverse phase HPLC retention times permit a further deconvolution of the library in terms of peptide size and hydrophobicity. Sequencing of cDNA derived using O-superfamily specific primers yielded five complete conotoxin precursor sequences, ranging in polypeptide length from 75-87 residues containing six Cys residues at the C terminus. Sequence analysis permits classification of the five putative mature peptides (Ac 6.1 to Ac 6.5) as delta, omega, and omega-like conotoxins. The presence of these predicted peptides in crude venom was established by direct matrix assisted laser desorption ionization tandem mass spectrometry (MALDI MS/MS) sequencing following trypsin digestion of the peptide mixture after global reduction/alkylation. The determination of partial peptide sequences and comparison with the predicted sequences resulted in the identification of four of the five predicted conotoxins. The characterization of posttranslationally modified analogs, which are hydroxylated at proline or amidated at the C-terminus is also demonstrated. Crude venom analysis should prove powerful in studying both inter- and intra-species variation in peptide libraries. PMID- 18286663 TI - Imatinib metabolite profiling in parallel to imatinib quantification in plasma of treated patients using liquid chromatography-mass spectrometry. AB - Besides affecting the systemic bioavailability of the parent drug, drug metabolizing enzymes (DMEs) may produce bioactive and/or toxic metabolites of clinical interest. We have investigated the capability to analyze simultaneously the parent drug and newly identified metabolites in patients' plasma by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The anticancer drug, imatinib, was chosen as a model drug because it has opened a new area in cancer therapy and is given orally and chronically. In addition, resistance and rare but sometimes severe side effects have been reported with this therapy. The quantification of imatinib and the profiling of its metabolites in plasma were established following three steps: (1) set-up of a generic sample extraction and LC-MS/MS conditions, (2) metabolite identification by LC-MS/MS using either in vitro incubations performed with human liver microsomes (HLMs) or patient plasma samples, (3) the simultaneous determination of plasma levels of imatinib and 14 metabolites in the plasma samples of 38 patients. Partial or cross method validation has been done and revealed that precise determinations of metabolite levels can be performed whereas pure standards are not available. Preliminary results indicate that the disposition of imatinib and its metabolites is related to interindividual variables and that outlier metabolite profiles can be revealed. This article underscores that, in addition to usual therapeutic drug monitoring (TDM), LC-MS/MS methods can simultaneously record a complete drug metabolic profile enabling various correlation studies of clinical interest. PMID- 18286664 TI - MALDI mass spectrometric determination of dendritic iron chelation stoichiometries and conditional affinity constants. AB - The iron chelation stoichiometries of a dendritic iron(III) chelator with N(1), N(3), N(5)-trimethylbenzene-1,3,5-tricarboxamide at its core, and containing 3 identical hexadentate tris-hydroxypyridinone branches D was studied by MALDI mass spectrometry. At pH 7.2, the speciation of the system included FeD, Fe(2)D and Fe(3)D species with the respective conditional stability constants of 26.74, 26.03 and 25.36. The differences in the stepwise affinity constants arise from the statistical distribution of iron(III), and there was no evidence for cooperativity between the iron-binding sites. PMID- 18286665 TI - Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) of low molecular weight organic compounds and synthetic polymers using zinc oxide (ZnO) nanoparticles. AB - We have developed surface-assisted laser desorption/ionization mass spectrometry using zinc oxide (ZnO) nanoparticles with anisotropic shapes (ZnO-SALDI-MS). The mass spectra showed low background noises in the low m/z, i.e. less than 500 u region. Thus, we succeeded in SALDI ionization on low molecular weight organic compounds, such as verapamil hydrochloride, testosterone, and polypropylene glycol (PPG) (average molecular weight 400) without using a liquid matrix or buffers such as citric acids. In addition, we found that ZnO-SALDI has advantages in post-source decay (PSD) analysis and produced a simple mass spectrum for phospholipids. The ZnO-SALDI spectra for synthetic polymers of polyethylene glycol (PEG), polystyrene (PS) and polymethylmethacrylate (PMMA) showed the sensitivity and molecular weight distribution to be comparable to matrix-assisted laser desorption/ionization (MALDI) spectra with a 2,5-dihydroxybenzoic acid (DHB) matrix. ZnO-SALDI shows good performance for synthetic polymers as well as low molecular weight organic compounds. PMID- 18286666 TI - Evaluation of anilinopyrimidine and other fungicides for control of grey mould (Botrytis cinerea) in container-grown Calluna vulgaris. AB - BACKGROUND: Grey mould (Botrytis cinerea Pers. ex Fr.) is a damaging disease affecting container-grown Calluna vulgaris (L.) Hull. Trials were conducted to test anilinopyrimidine fungicides (pyrimethanil, cyprodinil and mepanipyrim), azoxystrobin and tolylfluanid for crop safety on cultivars, and to evaluate spray programmes based on alternating these fungicides with different modes of action for disease control. Timing and application at different spray intervals were also investigated. RESULTS: Cyprodinil was slightly phytotoxic on only one of the seven cvs tested. Alternating nine-spray programmes of azoxystrobin with an anilinopyrimidine or tolylfluanid, or an anilinopyrimidine with tolylfluanid, with the first spray applied immediately after potting, were equally effective in reducing disease incidence and severity compared with untreated plants. Spray programmes of five or eight sprays, using an anilinopyrimidine and at least one other fungicide with a different mode of action, applied alternately or in blocks of two sprays of the same fungicide and immediately after potting, were equally effective in reducing disease. Omission of early fungicide sprays until 9 weeks after potting and subsequent alternate application of pyrimethanil and tolylfluanid with a different mode of action at 6 week intervals gave reduced disease control. CONCLUSION: There is potential to use spray programmes based on alternating anilinopyrimidines with fungicides with a different mode of action to provide effective control of B. cinerea on C. vulgaris. PMID- 18286667 TI - Direct injection of whole blood for liquid chromatography/tandem mass spectrometry analysis to support single-rodent pharmacokinetic studies. AB - Mass spectrometric developments in the last decade enable (sub)nanomolar detection of drug compounds in biological matrices in a few microliters of blood. However, the sampling and especially the handling of these small blood volumes is not straightforward. We studied the feasibility of a recently developed 'sorbent sampling technique' to handle these small blood volumes and the application to support pharmacokinetic (PK) screening programs. This technique applies 5-10 microL of blood on a fibrous material packed into a cartridge. Blood samples absorbed on these cartridges are eluted directly, on-line onto a solid-phase extraction liquid chromatography/tandem mass spectrometry (SPE-LC/MS/MS) system. It is shown that the sorbent sampling technique can be applied for a range of drug compounds. In spite of issues with recovery and sample clean-up that need further improvement, the sorbent sampling technique provided similar data as compared to conventional analytics. The technique was successfully applied to derive kinetic data from individual mice, thereby decreasing the number of required mice for a PK study from 21 to 3. PMID- 18286668 TI - Electrospray tandem mass spectrometric analysis of novel synthetic quinoxalinone derivatives. AB - Electrospray ionization tandem mass spectrometry (ESI-MS/MS) using a hybrid QqToF MS/MS instrument has aided the structural characterization and differentiation of a novel series of medicinal synthetic 1-N-glycoside-quinoxalinone derivatives. These derivatives 7 and 8 are formed by an amino bond between the cyclic N-1 of the quinoxaline moiety and the C-6 position of a fully protected methyl or allyl alpha-D-mannofuranoside 3 and 4, and subsequent deprotection of the mannopyranoside moiety. In general the novel synthetic quinoxaline derivatives afforded the protonated molecules in ESI. The breakdown routes of the protonated molecules were rationalized by conducting low-energy CID-MS/MS analyses. In addition, re-confirmation of the various established fragmentation routes was achieved by conducting a series of ESI-CID-QqTof-MS/MS product ion scans on various selected precursor ions, which were initiated by CID in the atmospheric pressure/vacuum interface using a higher declustering potential. ESI-QqToF-MS/MS analysis has proven to be a specific and very sensitive method for the structural identification in the gas phase of these novel glycoquinoxalinamine derivatives. PMID- 18286669 TI - Simultaneous profiling of multiple neurochemical pathways from a single cerebrospinal fluid sample using GC/MS/MS with electron capture detection. AB - Biogenic amines and amino acids are widely characterized in the pathways representing neurotransmission. Although several analytical methodologies have been used to detect specific target molecules in relevant fluids such as cerebrospinal fluid (CSF), multiple assays must be used to survey the primary pathways involved. This article describes the development of a GC/MS/MS method capable of analyzing up to 43 analytes (representing 20 amino acids and more than seven neurochemical pathways) from a single 50 microl CSF sample. In this procedure, a CSF sample is first treated with acetonitrile to precipitate proteins. The dried sample is then derivatized with a mixture of 2,2,3,3,3 pentafluoro-1-propanol and pentafluoropropionic acetic anhydride to replace all active hydrogen atoms with fluorine-containing groups. Due to the concentration difference between amino acids and neurotransmitters, these two compound classes are analyzed in separate injections of the same derivatized extract. The total run time for each injection is approximately 15-20 min. An essential feature of the method is the use of argon as a reagent gas for electron capture chemical ionization (ECCI), as the use of the more traditional gas (methane) lacked sufficient durability to be considered for use with the present instrumentation. This article describes the development of this method including a detailed investigation of the chemical ionization conditions used. The resultant conditions allow for the profiling of biogenic amines (e.g. serotonin, norepinephrine, and dopamine) in the low picogram per milliliter range. PMID- 18286670 TI - Substituted 3-phenylpropenoates and related analogs: electron ionization mass spectral fragmentation and density functional theory calculations. AB - Analysis of ethyl 3-(2-chlorophenyl)propenoate by electron ionization mass spectrometry showed the distinct loss of an ortho chlorine. To characterize the structural requisites for the observed mass fragmentation, a series of 30 halogen substituted 3-phenylpropenoate-related structures were examined. All ester containing alkene derivatives exhibited loss of the distinctive chlorine from the 2-position of the phenyl ring. Analogous derivatives with the halogen (chlorine or bromine) in the para position did not evidence selective halogen loss. Results demonstrated that substituted 3-phenylpropenoates and their analogs fragment via the formation of a previously reported benzopyrylium intermediate. To understand the correlation between the intramolecular radical substitution and the abundance and selectivity of the chlorine (or other halogen) displacement, density functional theory calculations were performed to determine the charge on the principal cation involved in the chlorine loss (in the ortho, meta, and para positions), the charge for the neutral radical (noncation), the excess alpha electron density on the relevant atom and the energy to form the cation from the neutral atom (ionization energy). Results showed that the selectivity and extent of halogen displacement correlated highly to the electrophilicity of the radical cation as well as the neutral radical. These data further support the proposed fragmentation mechanism involving intramolecular radical elimination. PMID- 18286671 TI - Derivatization of beta-dicarbonyl compound with 2,4-dinitrophenylhydrazine to enhance mass spectrometric detection: application in quantitative analysis of houttuynin in human plasma. AB - Houttuynin (decanoyl acetaldehyde), a beta-dicarbonyl compound, is the major antibacterial constituent in the volatile oil of Houttuynina cordata Thunb. In the present work, detection of houttuynin in human plasma based on the chemical derivatization with 2,4-dinitrophenylhydrazine (DNPH) coupled with liquid chromatography/tandem mass spectrometry was described. The primary reaction products between the beta-dicarbonyl compound and DNPH in aqueous phase were identified as heterocyclic structures, of which the mass spectrometric ionization and fragmentation behavior were characterized with the aid of high-resolution multistage mass spectral analysis. For quantification, houttuynin and internal standard (IS, benzophenone) in plasma were firstly converted to their DNPH derivatives without sample purification, then extracted from human plasma with n hexane and detected by liquid chromatography tandem mass spectrometry performed in selected reaction monitoring (SRM) mode. This method allowed for a lower limit of quantification (LLOQ) of 1.0 ng/ml using 100-microl plasma. The validation results showed high accuracy (%bias < 2.1) and precision (%CV < 7.2) at broad linear dynamic range (1.0-5000 ng/ml). The simple and quantitative derivatization coupled with tandem mass spectrometric analysis facilitates a sensitive and robust method for the determination of plasma houttuynin in pharmacokinetic studies. PMID- 18286672 TI - Outcome of fetuses with antenatally diagnosed short femur. AB - OBJECTIVES: To examine the outcome of fetuses diagnosed with short femur length at the time of the routine anomaly scan. METHODS: This was a retrospective review of all pregnancies referred to a tertiary referral unit with fetal femur length below the 5(th) centile for gestation at 18-24 weeks of gestation. All patients had undergone pregnancy dating and assessment of the risk of chromosomal abnormalities by measurement of fetal nuchal translucency at 11 to 13 + 6 weeks. RESULTS: Over 5 years, 129 cases were evaluated. Detailed ultrasound examination showed associated fetal abnormalities in 46 (36%) cases, and these were classified as non-isolated. In this group, skeletal dysplasias (n = 16), chromosomal abnormalities (n = 10) and genetic syndromes (n = 4) were the most common associations. In contrast, there were no cases of chromosomal abnormalities or skeletal dysplasia in the 83 (64%) isolated cases. Early severe intrauterine growth restriction (IUGR) with abnormal umbilical artery Doppler findings and delivery before 37 weeks occurred in 33/83 (40%) cases with isolated short femur, and 90% of these had abnormal uterine artery Doppler findings at the time of presentation. These pregnancies also had high rates of pre-eclampsia (36%) and intrauterine death (33%). Those with normal uterine artery Doppler imaging were at low risk for these complications. CONCLUSIONS: In a population previously screened by first-trimester fetal nuchal translucency measurement, the finding of isolated short femur at 18-24 weeks is unlikely to be due to aneuploidy. Severe IUGR associated with high mortality occurs in 40%, making uterine artery Doppler evaluation a useful clinical tool. PMID- 18286673 TI - Developing practical recommendations for the use of propensity scores: discussion of 'A critical appraisal of propensity score matching in the medical literature between 1996 and 2003' by Peter Austin, Statistics in Medicine. PMID- 18286674 TI - Comments on 'Statistical education for medical students--concepts are what remain when the details are forgotten' by Amir Herman, Netta Notzer, Zipi Libman, Rony Braunstein and David Steinberg. Statistics in Medicine 2007; 26:4344-4351. PMID- 18286675 TI - Neuro-regulation of lower esophageal sphincter function as treatment for gastroesophageal reflux disease. AB - The junction between the esophagus and the stomach is a specialized region, composed of lower esophageal sphincter (LES) and its adjacent anatomical structures, the gastric sling and crural diaphragm. Together these structures work in a coordinated manner to allow ingested food into the stomach while preventing reflux of gastric contents across the esophago-gastric junction (EGJ) into the esophagus. The same zone also permits retrograde passage of air and gastric contents into esophagus during belching and vomiting. The precise coordination required to execute such a complicated task is achieved by a finely regulated high-pressure zone. This zone keeps the junction between esophagus and stomach continuously closed, but is still able to relax briefly via input from inhibitory neurons that are responsible for its innervation. Alterations of the structure and function of the EGJ and the LES may predispose to gastroesophageal reflux disease (GERD). PMID- 18286676 TI - Venous thromboembolism with inflammatory bowel disease. AB - Venous thrombosis and thromboembolism appear to be increased in patients with inflammatory bowel disease. Although several acquired and genetic risk factors are known, about half that develop a thromboembolic event have no identifiable risk factor. Control of the inflammatory process is thought to be the key factor in risk reduction for thrombotic events. Prophylactic use of anticoagulants is not universally recommended, but possible use should be reviewed in an individual patient after evaluation of the risks, such as hemorrhage, compared to potential benefits. Particular consideration should be given if there has been a prior thrombotic event, if hospitalization will require surgery, or if an underlying coagulation disorder is present. PMID- 18286677 TI - Is acute recurrent pancreatitis a chronic disease? AB - Whether acute recurrent pancreatitis is a chronic disease is still debated and a consensus is not still reached as demonstrated by differences in the classification of acute recurrent pancreatitis. There is major evidence for considering alcoholic pancreatitis as a chronic disease ab initio while chronic pancreatitis lesions detectable in biliary acute recurrent pancreatitis (ARP) seem a casual association. Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation, hereditary and obstructive pancreatitis seem an acute disease that progress to chronic pancreatitis, likely as a consequence of the activation and proliferation of pancreatic stellate cells that produce and activate collagen and therefore fibrosis. From the diagnostic point of view, in patients with acute recurrent pancreatitis Endoscopic ultrasound (EUS) seems the more reliable technique for an accurate evaluation and follow-up of some ductal and parenchymal abnormalities suspected for early chronic pancreatitis. PMID- 18286679 TI - Diagnostic approach to patients with acute idiopathic and recurrent pancreatitis, what should be done? AB - Acute recurrent pancreatitis (ARP) is a common clinical condition that may be difficult to diagnose. Endoscopic ultrasound (EUS) is proposed to be a safe first line test of choice in the majority of patients. When interventions are needed to remove biliary stones, evaluate sphincter of Oddi or pancreas divisum, endoscopic retrograde cholangiopancreatography (ERCP) is recommended. Magnetic resonance cholangiopancreatography (MRCP) can be a suitable alternative from a diagnostic standpoint although may not be widely available. Finally, genetic testing is increasingly used to detect certain mutations that are associated with this diagnosis. PMID- 18286678 TI - Acute recurrent pancreatitis: an autoimmune disease? AB - In this review article, we will briefly describe the main characteristics of autoimmune pancreatitis and then we will concentrate on our aim, namely, evaluating the clinical characteristics of patients having recurrence of pain from the disease. In fact, the open question is to evaluate the possible presence of autoimmune pancreatitis in patients with an undefined etiology of acute pancreatitis and for this reason we carried out a search in the literature in order to explore this issue. In cases of recurrent attacks of pain in patients with "idiopathic" pancreatitis, we need to keep in mind the possibility that our patients may have autoimmune pancreatitis. Even though the frequency of this disease seems to be quite low, we believe that in the future, by increasing our knowledge on the subject, we will be able to diagnose an ever-increasing number of patients having acute recurrence of pain from autoimmune pancreatitis. PMID- 18286680 TI - Role of genetic disorders in acute recurrent pancreatitis. AB - There was remarkable progress in the understanding of the role genetic risk factors in chronic pancreatitis. These factors seem to be much more important than thought in the past. The rare autosomal-dominant mutations N29I and R122H of PRSS1 (cationic trypsinogen) as well as the variant N34S of SPINK1 (pancreatic secretory trypsin inhibitor) are associated to a disease onset in childhood or youth. Compared to chronic alcoholic pancreatitis the progression is slow so that for a long time only signs of acute-recurrent pancreatitis are found. Only at later time points (more than 10-15 years) there is evidence for chronic pancreatitis in the majority of patients. Acute recurrent pancreatitis may therefore be regarded as a transition state until definite signs of chronic pancreatitis are detectable. PMID- 18286681 TI - Endoscopic ultrasonography for evaluating patients with recurrent pancreatitis. AB - Acute recurrent pancreatitis (ARP) is still a complex diagnostic and therapeutic challenge in clinical practice. In up to 30% of cases of ARP, it is not possible to establish the etiology of the disease. In the other 70%, many factors play an etiological role in ARP: microlithiasis, sphincter of Oddi dysfunction (SOD), pancreas divisum, hereditary pancreatitis, cystic fibrosis, a choledochocele, annular pancreas, an anomalous pancreatobiliary junction, pancreatic tumors or chronic pancreatitis are diagnosed. EUS should be useful in ARP as it is sensitive for diagnosing bile duct stones, gallbladder sludge, pancreatic lesions, ductal abnormalities and chronic pancreatitis. Endoscopic ultrasound (EUS) appears to be diagnostic in the majority of patients with previously unexplained pancreatitis, and offers an alternative to endoscopic retrograde cholangiopancreatography (ERCP) as the initial diagnostic test in patients with ARP. PMID- 18286682 TI - Sphincter of Oddi dysfunction and bile duct microlithiasis in acute idiopathic pancreatitis. AB - Although there are numerous causes of acute panc-reatitis, an etiology cannot always be found. Two potential etiologies, microlithiasis and sphincter of Oddi dysfunction, are discussed in this review. Gallbladder microlithiasis, missed on transcutaneous ultrasound, is reported as the cause of idiopathic acute pancreatitis in a wide frequency range of 6%-80%. The best diagnostic technique for gallbladder microlithiasis is endoscopic ultrasound although biliary crystal analysis and empiric cholecystectomy remain as reasonable options. In contrast, in patients who are post-cholecystectomy, bile duct microlithiasis does not appear to have a role in the pathogenesis of acute pancreatitis. Sphincter of Oddi dysfunction is present in 30%-65% of patients with idiopathic acute recurrent pancreatitis in whom other diagnoses have been excluded. It is unclear if this sphincter dysfunction was the original etiology of the first episode of pancreatitis although it appears to have a causative role in recurring episodes since sphincter ablation decreases the frequency of recurrent attacks. Unfortunately, this conclusion is primarily based on small retrospective case series; larger prospective studies of the outcome of pancreatic sphincterotomy for SOD-associated acute pancreatitis are sorely needed. Another problem with this diagnosis and its treatment is the concern over potential procedure related complications from endoscopic retrograde cholangiopancreatography (ERCP), manometry and pancreatic sphincterotomy. For these reasons, patients should have recurrent acute pancreatitis, not a single episode, and have a careful informed consent before assessment of the sphincter of Oddi is undertaken. PMID- 18286683 TI - Pancreatic ductal system obstruction and acute recurrent pancreatitis. AB - Acute recurrent pancreatitis is a clinical entity largely associated with pancreatic ductal obstruction. This latter includes congenital variants, of which pancreas divisum is the most frequent but also controversial, chronic pancreatitis, tumors of the pancreaticobiliary junction and sphincter of Oddi dysfunction. This review summarizes current knowledge about diagnostic work-up and therapy of these conditions. PMID- 18286684 TI - Endoscopic therapy in acute recurrent pancreatitis. AB - Endoscopic retrograde cholangiopancreatography (ERCP) has evolved from a largely diagnostic to a largely therapeutic modality. Cross-sectional imaging, such as computed tomography (CT) and magnetic resonance imaging (MRI), and less invasive endoscopy, especially endoscopic ultrasound (EUS), have largely taken over from ERCP for diagnosis. However, ERCP remains the "first line" therapeutic tool in the management of mechanical causes of acute recurrent pancreatitis, including bile duct stones (choledocholithiasis), ampullary masses (benign and malignant), congenital variants of biliary and pancreatic anatomy (e.g. pancreas divisum, choledochoceles), sphincter of Oddi dysfunction (SOD), pancreatic stones and strictures, and parasitic disorders involving the biliary tree and/or pancreatic duct (e.g Ascariasis, Clonorchiasis). PMID- 18286685 TI - Diagnosis and management of relapsing pancreatitis associated with cystic neoplasms of the pancreas. AB - One of the most important causes of relapsing pancreatitis is a cystic neoplasm of the pancreas. These low grade malignancies may cause pancreatitis by obstructing or communicating with a pancreatic duct. Patients with relapsing pancreatitis and a focal fluid fluid collection should be investigated for the possibility of a mucinous cystic neoplasm. Cross sectional imaging can provide a diagnosis with the imaging findings of a low attenuation cystic lesion containing mural calcification (CT scanning) or a lobular T2 enhancing lesion (MRCP). Endoscopic ultrasound can provide more detailed imaging with the ability to guide fine needle aspiration of the cyst fluid. Cyst fluid analysis can provide a diagnosis of a mucinous cystic lesion with the combination of cytology (mucinous epithelium), elevated carcinoembryonic antigen (CEA), and the presence of DNA mutations. Management of these patients consists of surgical resection and monitoring in patients not able to withstand surgery. PMID- 18286686 TI - Overexpression of Slug is associated with malignant progression of esophageal adenocarcinoma. AB - AIM: To characterise expression of known E-cadherin repressors; Snail, Slug and Twist in the development of esophageal adenocarcinoma. METHODS: E-cadherin, Slug, Snail and Twist mRNA expression in Barrett's metaplasia and esophageal adenocarcinoma specimens was examined by real-time reverse transcription polymerase chain reaction (RT-PCR). Semi-quantitative immunohistochemistry was used to examine cellular localisation and protein levels. The effect of Slug on epithelial mesenchymal transition (EMT) markers was examined by transfection of Slug into an adenocarcinoma line OE33. RESULTS: Cellular localisation of Slug in Barrett's metaplasia was largely cytoplasmic whilst in adenocarcinoma it was nuclear. Semi-quantitative analysis indicated that Slug was more abundant in adenocarcinoma compared to matched Barrett's metaplastic specimens. Snail and Twist were expressed in adenocarcinoma but were cytoplasmic in location and not induced compared to Barrett's mucosa. These observations were supported by mRNA studies where only Slug mRNA was shown to be over-expressed in adenocarcinoma and inversely correlated to E-cadherin expression. Overexpression of Slug in OE33 mediated E-cadherin repression and induced the mesenchymal markers vimentin and fibronectin. CONCLUSION: Progression to adenocarcinoma is associated with increased Slug expression and this may represent a mechanism of E-cadherin silencing. PMID- 18286687 TI - Frequent mutations of the CA simple sequence repeat in intron 1 of EGFR in mismatch repair-deficient colorectal cancers. AB - AIM: To investigate the polymorphic simple sequence repeat in intron 1 of the epidermal growth factor receptor gene (EGFR) (CA-SSR I), which is known to affect the efficiency of gene transcription as a putative target of the mismatch repair (MMR) machinery in colorectal tumors. METHODS: The CA-SSR I genotype was analyzed in a total of 86 primary colorectal tumors, selected upon their microsatellite instability (MSI) status [42 with high frequency MSI (MSI-H) and 44 microsatellite stable (MSS)] and their respective normal tissue. The effect of the CA-SSR I genotype on the expression of the EGFR gene was evaluated in 18 specimens using quantitative real-time reverse transcription PCR and immunohistochemistry. RESULTS: Mutations in CA-SSR I were detected in 86% (36 of 42) of MSI-H colorectal tumors and 0% (0 of 44) of MSS tumors, indicating the EGFR gene as a novel putative specific target of the defective MMR system (P < 0.001). Impaired expression of EGFR was detected in most of the colorectal tumors analyzed [6/12 (50%) at the mRNA level and 15/18 (83%) at the peptide level]. However, no association was apparent between EGFR expression and CA-SSR I status in tumors or normal tissues. CONCLUSION: Our results suggest that CA-SSR I sequence does not contribute to the regulation of EGFR transcription in colon, and should thus not be considered as a promising predictive marker for response to EGFR inhibitors in patients with colorectal cancer. PMID- 18286688 TI - Predictive value of Ki67 and p53 in locally advanced rectal cancer: correlation with thymidylate synthase and histopathological tumor regression after neoadjuvant 5-FU-based chemoradiotherapy. AB - AIM: To investigate the predictive value of Ki67 and p53 and their correlation with thymidylate synthase (TS) gene expression in a rectal cancer patient cohort treated according to a standardized recommended neoadjuvant treatment regimen. METHODS: Formalin fixed, paraffin embedded pre-therapeutical tumor biopsies (n = 22) and post-therapeutical resection specimens (n = 40) from patients with rectal adenocarcinoma (clinical UICC stage II/III) receiving standardized neoadjuvant 5 fluorouracil (5-FU) based chemoradiotherapy were studied for Ki67 and p53 expression by immunohistochemistry and correlated with TS mRNA expression by quantitative TaqMan real-time PCR after laser microdissection. The results were compared with histopathological tumor regression according to a standardized semiquantitative score grading system. RESULTS: Responders (patients with high tumor regression) showed a significantly lower Ki67 expression than non responders in the pre-therapeutical tumor biopsies (81.2% vs 16.7%; P < 0.05) as well as in the post-therapeutical resection specimens (75.8% vs 14.3%; P < 0.01). High TS mRNA expression was significantly correlated with a high Ki67 index and low TS mRNA expression was significantly correlated with a low Ki67 index in the pre-therapeutical tumor biopsies (corr. coef. = 0.46; P < 0.01) as well as in the post-therapeutical resection specimens (corr. coef. = 0.40; P < 0.05). No significant association was found between p53 and TS mRNA expression or tumor regression. CONCLUSION: Ki67 has, like TS, predictive value in rectal cancer patients after neoadjuvant 5-FU based chemoradiotherapy. The close correlation between Ki67 and TS indicates that TS is involved in active cell cycle processes. PMID- 18286689 TI - Infant intestinal Enterococcus faecalis down-regulates inflammatory responses in human intestinal cell lines. AB - AIM: To investigate the ability of Lactic acid bacteria (LAB) to modulate inflammatory reaction in human intestinal cell lines (Caco-2, HT-29 and HCT116). Different strains of LAB isolated from new born infants and fermented milk, together with the strains obtained from culture collections were tested. METHODS: LABs were treated with human intestinal cell lines. ELISA was used to detect IL-8 and TGF-beta protein secretion. Cytokines and Toll like receptors (TLRs) gene expression were assessed using RT-PCR. Conditional medium, sonicated bacteria and UV killed bacteria were used to find the effecter molecules on the bacteria. Carbohydrate oxidation and protein digestion were applied to figure out the molecules' residues. Adhesion assays were further carried out. RESULTS: It was found that Enterococcus faecalis is the main immune modulator among the LABs by downregulation of IL-8 secretion and upregulation of TGF-beta. Strikingly, the effect was only observed in four strains of E. faecalis out of the 27 isolated and tested. This implies strain dependent immunomodulation in the host. In addition, E. faecalis may regulate inflammatory responses through TLR3, TLR4, TLR9 and TRAF6. Carbohydrates on the bacterial cell surface are involved in both its adhesion to intestinal cells and regulation of inflammatory responses in the host. CONCLUSION: These data provide a case for the modulation of intestinal mucosal immunity in which specific strains of E. faecalis have uniquely evolved to maintain colonic homeostasis and regulate inflammatory responses. PMID- 18286690 TI - Sources of calcium in agonist-induced contraction of rat distal colon smooth muscle in vitro. AB - AIM: To study the origin of calcium necessary for agonist-induced contraction of the distal colon in rats. METHODS: The change in intracellular calcium concentration ([Ca2+]i) evoked by elevating external Ca2+ was detected by fura 2/AM fluorescence. Contractile activity was measured with a force displacement transducer. Tension was continuously monitored and recorded using a Powerlab 4/25T data acquisition system with an ML110 bridge bioelectric physiographic amplifier. RESULTS: Store depletion induced Ca2+ influx had an effect on [Ca2+]i. In nominally Ca2+-free medium, the sarco-endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin (1 mumol/L) increased [Ca2+]i from 68 to 241 nmol/L, and to 458 (P < 0.01) and 1006 nmol/L (P < 0.01), respectively, when 1.5 mmol/L and 3.0 mmol/L extracellular Ca2+ was reintroduced. Furthermore, the change in [Ca2+]i was observed with verapamil (5 micromol/L), La3+ (1 mmol/L) or KCl (40 mmol/L) in the bathing solution. These channels were sensitive to La3+ (P < 0.01), insensitive to verapamil, and voltage independent. In isolated distal colons we found that in normal Krebs solution, contraction induced by acetylcholine (ACh) was partially inhibited by verapamil, and the inhibitory rate was 41% (P < 0.05). On the other hand, in Ca2+-free Krebs solution, ACh induced transient contraction due to Ca2+ release from the intracellular stores. The transient contraction lasted until the Ca2+ store was depleted. Restoration of extracellular Ca2+ in the presence of atropine produced contraction, mainly due to Ca2+ influx. Such contraction was not inhibited by verapamil, but was decreased by La3+ (50 micromol/L) from 0.96 to 0.72 g (P < 0.01). CONCLUSION: The predominant source of activator Ca2+ for the contractile response to agonist is extracellular Ca2+, and intracellular Ca2+ has little role to play in mediating excitation-contraction coupling by agonists in rat distal colon smooth muscle in vitro. The influx of extracellular Ca2+ is mainly mediated through voltage-, receptor- and store-operated Ca2+ channels, which can be used as an alternative to develop new drugs targeted on the dysfunction of digestive tract motility. PMID- 18286691 TI - Factors associated with time to laparoscopic cholecystectomy for acute cholecystitis. AB - AIM: To determine patient and process of care factors associated with performance of timely laparoscopic cholecystectomy for acute cholecystitis. METHODS: A retrospective medical record review of 88 consecutive patients with acute cholecystitis was conducted. Data collected included demographic data, co morbidities, symptoms and physical findings at presentation, laboratory and radiological investigations, length of stay, complications, and admission service (medical or surgical). Patients not undergoing cholecystectomy during this hospitalization were excluded from analysis. Hierarchical generalized linear models were constructed to assess the association of pre-operative diagnostic procedures, presenting signs, and admitting service with time to surgery. RESULTS: Seventy cases met inclusion and exclusion criteria, among which 12 were admitted to the medical service and 58 to the surgical service. Mean +/- SD time to surgery was 39.3 +/- 43 h, with 87% of operations performed within 72 h of hospital arrival. In the adjusted models, longer time to surgery was associated with number of diagnostic studies and endoscopic retrograde cholangio pancreatography (ERCP, P = 0.01) as well with admission to medical service without adjustment for ERCP (P < 0.05). Patients undergoing both magnetic resonance cholangiopancreatography (MRCP) and computed tomography (CT) scans experienced the longest waits for surgery. Patients admitted to the surgical versus medical service underwent surgery earlier (30.4 +/- 34.9 vs 82.7 +/- 55.1 h, P < 0.01), had less post-operative complications (12% vs 58%, P < 0.01), and shorter length of stay (4.3 +/- 3.4 vs 8.1 +/- 5.2 d, P < 0.01). CONCLUSION: Admission to the medical service and performance of numerous diagnostic procedures, ERCP, or MRCP combined with CT scan were associated with longer time to surgery. Expeditious performance of ERCP and MRCP and admission of medically stable patients with suspected cholecystitis to the surgical service to speed up time to surgery should be considered. PMID- 18286692 TI - An easier method for performing a pancreaticojejunostomy for the soft pancreas using a fast-absorbable suture. AB - AIM: To clarify the usefulness of a new method for performing a pancreaticojejunostomy by using a fast-absorbable suture material irradiated polyglactin 910, and a temporary stent tube for a narrow pancreatic duct with a soft pancreatic texture. METHODS: Among 63 consecutive patients with soft pancreas undergoing a pancreaticoduodenectomy from 2003 to 2006, 35 patients were treated with a new reconstructive method. Briefly, after the pancreatic transaction, a stent tube was inserted into the lumen of the pancreatic duct and ligated with it by a fast-absorbable suture. Another tip of the stent tube was introduced into the intestinal lumen at the jejunal limb, where a purse-string suture was made by another fast-absorbable suture to roughly fix the tube. The pancreaticojejunostomy was completed by ligating two fast-absorbable sutures to approximate the ductal end and the jejunal mucosa, and by adding a rough anastomosis between the pancreatic parenchyma and the seromuscular layer of the jejunum. The initial surgical results with this method were retrospectively compared with those of the 28 patients treated with conventional duct-to-mucosa anastomosis. RESULTS: The incidences of postoperative morbidity including pancreatic fistula were comparable between the two groups (new; 3%-17% vs conventional; 7%-14% according to the definitions). There was no mortality and re admission. Late complications were also rarely seen. CONCLUSION: A pancreaticojejunostomy using an irradiated polyglactin 910 suture material and a temporary stent is easy to perform and is feasible even in cases with a narrow pancreatic duct and a normal soft pancreas. PMID- 18286693 TI - Endoscopic transpapillary brush cytology and forceps biopsy in patients with hilar cholangiocarcinoma. AB - AIM: To evaluate the sensitivity of brush cytology and forceps biopsy in a homogeneous patient group with hilar cholangiocarcinoma. METHODS: Brush cytology and forceps biopsy were routinely performed in patients with suspected malignant biliary strictures. Fifty-eight consecutive patients undergoing endoscopic retrograde cholangio-pancreatography (ERCP) including forceps biopsy and brush cytology in patients with hilar cholangiocarcinoma between 1995-2005. RESULTS: Positive results for malignancy were obtained in 24/58 patients (41.4%) by brush cytology and in 31/58 patients (53.4%) by forceps biopsy. The combination of both techniques brush cytology and forceps biopsy resulted only in a minor increase in diagnostic sensitivity to 60.3% (35/58 patients). In 20/58 patients (34.5%), diagnosis were obtained by both positive cytology and positive histology, in 11/58 (19%) by positive histology (negative cytology) and only 4/58 patients (6.9%) were confirmed by positive cytology (negative histology). CONCLUSION: Brush cytology and forceps biopsy have only limited sensitivity for the diagnosis of malignant hilar tumors. In our eyes, additional diagnostic techniques should be evaluated and should become routine in patients with negative cytological and histological findings. PMID- 18286694 TI - Effect of preoperative biliary drainage on surgical results after pancreaticoduodenectomy in patients with distal common bile duct cancer: focused on the rate of decrease in serum bilirubin. AB - AIM: To examine if the rate of decrease in serum bilirubin after preoperative biliary drainagecan be used as a predicting factor for surgical complications and postoperative recovery after pancreaticoduodenectomy in patients with distal common bile duct cancer. METHODS: A retrospective study was performed in 49 consecutive patients who underwent pancreaticoduodenectomy for distal common bile duct cancer. Potential risk factors were compared between the complicated and uncomplicated groups. Also, the rates of decrease in serum bilirubin were compared pre- and postoperatively. RESULTS: Preoperative biliary drainage (PBD) was performed in 40 patients (81.6%). Postoperative morbidity and mortality rates were 46.9% (23/49) and 6.1% (3/49), respectively. The presence or absence of PBD was not different between the complicated and uncomplicated groups. In patients with PBD, neither the absolute level nor the rate of decrease in serum bilirubin was significantly different. Patients with rapid decrease preoperatively showed faster decrease during the first postoperative week (5.5 +/- 4.4 micromol/L vs 1.7 +/- 9.9 micromol/L, P = 0.004). CONCLUSION: PBD does not affect the surgical outcome of pancreaticoduodenectomy in patients with distal common bile duct cancer. There is a certain group of patients with a compromised hepatic excretory function, which is represented by the slow rate of decrease in serum bilirubin after PBD. PMID- 18286695 TI - Ursodeoxycholic acid treatment improves hepatocyte ultrastructure in rat liver fibrosis. AB - AIM: To examine the ultrastructural changes after ursodeoxycholic acid (UDCA) treatment in hepatocytes from experimentally induced fibrotic livers. METHODS: Liver fibrosis was induced in male Sprague-Dawley rats with CCl4 for 12 wk, and the rats were divided into two groups. Group I was treated with saline and group II with UDCA (25 mg/kg per day) for 4 wk. All the rats were killed at wk 16. Mitochondria, nuclei, rough endoplasmic reticulum (RER) and smooth endoplasmic reticulum (SER) of hepatocytes were evaluated according to a scoring system. RESULTS: Mitochondria, nuclei, RER and SER injury scores in group II were significantly lower than those in group I (P < 0.001). CONCLUSION: UDCA alleviates hepatocyte organelle injury in CCl4-induced liver fibrosis. PMID- 18286696 TI - Effect of viral load on T-lymphocyte failure in patients with chronic hepatitis B. AB - AIM: To investigate peripheral T-lymphocyte sub-population profile and its correlation with hepatitis B virus (HBV) replication in patients with chronic hepatitis B (CHB). METHODS: Distribution of T-lymphocyte subpopulations in peripheral blood was measured by flow cytometry in 206 CHB patients. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real time polymerase chain reaction (PCR). The relationship between HBV replication and variation in peripheral T-cell subsets was analyzed. RESULTS: CHB patients had significantly decreased CD3+ and CD4+ cells and CD4+/CD8+ ratio, and increased CD8+ cells compared with uninfected controls (55.44 +/- 12.39 vs 71.07 +/- 4.76, 30.92 +/- 7.48 vs 38.94 +/- 3.39, 1.01 +/- 0.49 vs 1.67 +/- 0.33, and 34.39 +/- 9.22 vs 24.02 +/- 4.35; P < 0.001, respectively). Univariate analysis showed a similar pattern of these parameters was significantly associated with high viral load, presence of serum hepatitis B e antigen (HBeAg) expression, liver disease severity, history of maternal HBV infection, and young age at HBV infection, all with P < 0.01. There was a significant linear relationship between viral load and these parameters of T-lymphocyte subpopulations (linear trend test P < 0.001). There was a negative correlation between the levels of CD3+ and CD4+ cells and CD4+/CD8+ ratio and serum level of viral load in CHB patients (r = 0.68, -0.65 and -0.75, all P < 0.0001), and a positive correlation between CD8+ cells and viral load (r = 0.70, P < 0.0001). There was a significant decreasing trend in CD3+ and CD4+ cells and CD4+/CD8+ ratio with increasing severity of hepatocyte damage and decreasing age at HBV infection (linear trend test P < 0.01). In multiple regression (after adjustment for age at HBV infection, maternal HBV infection status and hepatocyte damage severity) log copies of HBV DNA maintained a highly significant predictive coefficient on T-lymphocyte subpopulations, and was the strongest predictor of variation in CD3+, CD4+, CD8+ cells and CD4+/CD8+ ratio. However, the effect of HBeAg was not significant. CONCLUSION: T-lymphocyte failure was significantly associated with viral replication level. The substantial linear dose-response relationship and strong independent predictive effect of viral load on T-lymphocyte subpopulations suggests the possibility of a causal relationship between them, and indicates the importance of viral load in the pathogenesis of T cell hyporesponsiveness in these patients. PMID- 18286698 TI - KAI1 is a potential target for anti-metastasis in pancreatic cancer cells. AB - AIM: To investigate whether KAI1, as a metastasis suppressor gene, is associated with invasive and metastatic ability of pancreatic cancer cells. METHODS: KAI1 gene was transfected into pancreatic cancer cell line MiaPaCa II by liposomes selected with G418. Expression of transfected cells was measured by Western blotting, immunofluorescence and immunocytochemistry. Tumor cell invasion and metastatic ability were detected through gelatinase activity and reconstituted basement membrane (Matrigel) assay. pCMV-KAI1 was directly injected into the heterotopic human pancreatic adenocarcinoma successfully established in the groin of BALB/C nude mice, by subcutaneous injection of MiaPaCa II pancreatic cancer cells. The statistical analysis between groups was determined by Student's two tailed t test. RESULTS: By Western blotting, MiaPaCa II cells transfected by KAI1 gene indicated KAI1 expression at approximately 29.1 kDa. Cytoplasm staining was positive and uniformly spread in transfected cancer cells, using immunohistochemistry and immunofluorescence. The most obvious difference was present after 30 h (MiaPaca II 43.6 +/- 9.42, pCMV-MiaPaca II 44.8 +/- 8.56, pCMV KAI1-MiaPaca II 22.0 +/- 4.69, P < 0.05). Gelatinolysis revealed a wider and clearer band of gelatinolytic activity in non-transfected than in transfected cells (MiaPaCa II cells 30.8 +/- 0.57, transfected cells 28.1 +/- 0.65, P < 0.05). In vivo tumor growth rates of KAI1 transfectants with KAI1-Lipofectamine 1.22 +/- 0.31 in A group were lower than control 4.61 +/- 1.98 and pCMV-KAI 11.67 +/- 0.81. Analyses of metastases with and without KAI1 transfection in mice were different in liver and lung between controls 1.62 +/- 0.39, 0.45 +/- 0.09, pCMV KAI 1.01 +/- 0.27, 0.33 +/- 0.09 and KAI1-Lipofectamine 0.99 +/- 0.21, 0.30 +/- 0.09 respectively (P < 0.05). CONCLUSION: High expression of KAI1 gene was found in transfected MiaPaCa II human pancreatic cancer cells with lower metastatic ability. KAI1 gene plays an important role in inhibiting metastasis of pancreatic cancer after direct injection into pancreatic adenocarcinoma. These results show that the suppressed invasion and motor function of pancreatic cancer cells may be a key reason why the KAI1 gene controls pancreatic cancer cell metastasis. PMID- 18286697 TI - Comparative analysis of intestinal microbial community diversity between healthy and orally infected ducklings with Salmonella enteritidis by ERIC-PCR. AB - AIM: To analyze the difference of intestinal microbial community diversity between healthy and (S. enteritidis) orally infected ducklings. METHODS: Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR was applied to analyze the intestinal microbial community diversity and dynamic change including duodenum, jejunum, ileum, cecum and rectum from healthy ducklings and 7-day-old ducklings after oral infection with S. enteritidis at different time points. RESULTS: The intestinal microbial community of the control healthy ducklings was steady and the ERIC-PCR band numbers of the control healthy ducklings were the least with rectum and were the most with caecum. ERIC-PCR bands of orally inoculated ducklings did not obviously change until 24 h after inoculation (p.i.). The numbers of the ERIC-PCR bands gradually decreased from 24 h to 72 h p.i., and then, with the development of disease, the band numbers gradually increased until 6 d p.i. The prominent bacteria changed because of S. enteritidis infection and the DNAstar of staple of ERIC-PCR showed that aerobe and facultative aerobe (Escherichia coli, Shigella, Salmonella) became preponderant bacilli in the intestine of orally infected ducklings with SE. CONCLUSION: This study has provided significant data to clarify the intestinal microbial community diversity and dynamic change of healthy and S. enteritidis orally infected ducklings, and valuable insight into the pathogenesis of S. enteritidis infection in both human and animals. PMID- 18286699 TI - Laparoscopic hepatic left lateral lobectomy combined with fiber choledochoscopic exploration of the common bile duct and traditional open operation. AB - AIM: To investigate the possibilities and advantages of laparoscopic hepatic left lateral lobectomy combined with fiber choledochoscopic exploration of the common bile duct compaired with traditional open operation. METHODS: Laparoscopic hepatic left lateral lobectomy combined with fiber choledochoscopic exploration of the common bile duct and traditional open operation were performed in two groups of patients who had gallstones in the left lobe of liver and in the common bile duct. The hospitalization time, hospitalization costs, operation time, operative complications and post-operative liver functions of the two groups of patients were studied. RESULTS: The operation time and post-operative liver functions of the two groups of patients had no significant differences, while the hospitalization time, hospitalization costs and operative complications of the laparoscopic hepatic left lateral lobectomy combined with fiber choledochoscopic exploration in the common bile duct group were significantly lower than those in the traditional open operation group. CONCLUSION: For patients with gallstones in the left lobe of liver and in the common bile duct, laparoscopic hepatic left lateral lobectomy combined with fiber choledochoscopic exploration of the common bile duct can significantly shorten the hospitalization time, reduce the hospitalization costs and the post-operative complications, without prolonging the operation time and bringing about more liver function damages compared with traditional open operation. This kind of operation has more advantages than traditional open operation. PMID- 18286700 TI - Clear cell adenocarcinoma of the colon: a case report and review of literature. AB - A primary clear cell adenocarcinoma of the colon is a rare oncologic entity. We herein report a case of such a tumor of the sigmoid colon in a 71-year-old woman who was successfully treated by an endoscopic polypectomy in our hospital. We also reviewed the published reports regarding cases of primary clear cell tumors in the colon. PMID- 18286702 TI - Collaborative research from a methodological point of view. Editorial introduction. PMID- 18286701 TI - Rapunzel syndrome: the unsuspected culprit. AB - Trichobezoar is a rare intriguing disorder in which swallowed hairs accumulates in the stomach. Being indigestible and slippery, it could not be propulsed and becomes entrapped within the stomach. Large amounts can thus accumulate over the years forming a hair ball. Rapunzel syndrome is a variant where hair accumulation reaches the small gut and beyond in some cases. Although the syndrome has been known for many years, only 24 cases have been reported in the literature and the discovery of a new case is always surprising. In this report, we present two cases discovered within a period of three months. One of them was pregnant and had small bowel intussusception and perforation, a very rare combination. We hereby add two more cases to the literature. To our knowledge, this is the first report on two cases of Rapunzel syndrome, the diagnosis of which demands a high index of suspicion. PMID- 18286703 TI - Overview of conference on preclinical abuse liability testing: current methods and future challenges. PMID- 18286704 TI - Summary of NIDA medications workshop: new opportunities for chemists and pharmacologists. PMID- 18286705 TI - Nicotine addiction: past, present and future. Marian Fischman lecture given at the 2007 meeting of CPDD. PMID- 18286707 TI - A winding road towards an HIV vaccine. PMID- 18286708 TI - Proceedings of the 10th International Symposium on the Treatment of Parkinson's Disease, Kobe, Japan, 2005. PMID- 18286709 TI - UK Government considers supervised injecting clinics. PMID- 18286710 TI - Nepalese women under the shadow of domestic violence. PMID- 18286711 TI - History as a medical tool. PMID- 18286712 TI - Levetiracetam has no acute effects on brain gamma-aminobutyric acid levels. AB - OBJECTIVE: The mechanism of action of levetiracetam (LEV), an antiepileptic drug, is related to a novel binding site, SV2, but LEV acts on GABA-A receptors. The objective of the study described here was to determine if LEV modulates brain GABA in vivo. METHODS: Concentrations of cerebral GABA and serum LEV were obtained in seven healthy individuals using 1H magnetic resonance spectroscopy at baseline and 3 and 6 hours following oral administration of 1 g of LEV. RESULTS: Brain cerebral GABA acutely concentrations did not change from baseline. CONCLUSION: The results indicate that LEV does not increase human cerebral GABA concentrations acutely in healthy individuals. PMID- 18286713 TI - Depression in a community-dwelling sample of older adults with late-onset or lifetime epilepsy. AB - Epilepsy is one of the most common neurological disorders of late adulthood, yet little research has examined the impact of epilepsy in an older population, particularly in relation to depression. It has been argued that the symptomatology of interictal depression closely resembles that of dysthymia; however, our understanding of depression in older people with epilepsy is generalized from younger populations. This study examined the prevalence and symptomatology of depression, using both self-report and clinical interview, in a community-dwelling group of older adults with epilepsy compared with an age matched control group. An additional aim of the study was to determine if a self reported history of depression was a predictor of late-onset (after 55 years) epilepsy. Among the epilepsy group, 40.6% reported symptoms of depression that were congruent with dysthymic-like disorder of epilepsy; however, no relationship was apparent between prior depression and late-onset epilepsy. PMID- 18286714 TI - Retraction notice to "Structure and functional analysis of the MYND domain" [J. Mol. Biol. (2006) 358, 498-508]. PMID- 18286715 TI - Heart death rates show a nation of inequalities. PMID- 18286716 TI - Royal Marsden blaze. Cool heads and smart systems stop trust care combusting. PMID- 18286717 TI - Spot the difference. PMID- 18286718 TI - Improvement culture. Fantastic four can make superheroes of us all. PMID- 18286719 TI - Data briefing. Waiting times: cutting inequality. PMID- 18286720 TI - Brown unveils vision for 21st century health. PMID- 18286721 TI - Acute care. Battle of the sexes: the mixed ward row that won't lie down. PMID- 18286722 TI - Managing cultures. PMID- 18286723 TI - Comparing reforms. Home truths from the other side of the world. PMID- 18286724 TI - Data briefing. Sixty years of eating and smoking. PMID- 18286725 TI - First-hand history. In at the start. PMID- 18286726 TI - National IT programme. Strike the right balance. Interview by Stuart Shepherd. PMID- 18286727 TI - A nine session manual of motivational enhancement therapy for methamphetamine dependence: adherence and efficacy. AB - Motivational enhancement therapy (MET) is a brief therapy shown to be effective for problem drinkers. Because the response to MET for other addictive disorders is mixed, we assessed the utility of increasing the number of sessions in subjects with methamphetamine (MA) dependence. One therapist was trained in a nine-session manual of MET, which was tested over eight weeks in 30 MA-dependent outpatients. Adherence to the manual was assessed by two raters, who reviewed a random sample of 15 audiotaped therapy sessions. Interventions were rated on a seven-point Likert scale for frequency/extensiveness (1 = not at all to 7 = extensively) and skill level (1 = unacceptable to 7 = high level of mastery). Ratings of adherence were moderate for frequency/extensiveness (4.2 +/- 2.2 and 4.3 +/- 1.8; Mean +/- SD) and high for skill level (5.4 +/- 0.6 and 5.2 +/- 0.4). Subjects attended 7.0 +/- 2.5 (78%) of nine sessions. Self-reported days of methamphetamine use decreased from 841/1793 (47%) of the 60 days prior to study entry to 448/1458 (31%) during the study (p = 0.011). MA-positive urine samples decreased from 76/118 (64%) during screening to 93/210 (44%) during treatment (p = 0.015). The MET manual was readily learned, and subjects attended a high proportion of therapy sessions with marked reductions in methamphetamine use. PMID- 18286728 TI - New drug combo lowers blood pressure: all diabetics benefit. Two drugs offer lifesaving benefits, even in diabetes with normal blood pressure. PMID- 18286729 TI - Good patient-doctor communication is vital. The healing process can be viewed as a doctor-patient partnership, facilitated and enhanced by good interaction. PMID- 18286730 TI - Diagnosing and treating H. Pylori. There is significant new information on this common intestinal infection's diagnosis and treatment. PMID- 18286731 TI - Combat loneliness to slow the aging process. Feeling isolated could accelerate aging, affecting inflammation and infection at the cellular level. PMID- 18286732 TI - New annual osteoporosis drug reduces risk of death, future fractures. Infused dose is more convenient for patients, enhances overall quality of life. PMID- 18286733 TI - Does the advice about lowering high blood pressure (e.g., eat a low-salt diet, control your weight) apply to people whose high blood pressure is already controlled by medication? PMID- 18286734 TI - [Cytomegalovirus infection in patients with renal transplantation]. PMID- 18286735 TI - [Echovirus18 (Echo18) outbreak in an intensive care unit for newborn infants]. PMID- 18286736 TI - [Therapeutic effects of anti-influenza drugs]. PMID- 18286737 TI - [The Fondiaria insurance society and the birth of the INA]. PMID- 18286738 TI - [A historic excommunication]. PMID- 18286739 TI - Tribulations and achievements: the early history of Olympism in Argentina. AB - By the end of the nineteenth century, modern sport had enchanted the people of Argentina. At that time the nation enjoyed a remarkable degree of economic prosperity and embarked on increasing political democratization. These circumstances, along with the fact that the nation was represented from the beginning, in 1894, on the International Olympic Committee seemed to favour Argentina as the spearhead of the diffusion of Olympism throughout South America. However, the country only enjoyed its first official Olympic participation in the Paris Games of 1924 - a few months after the establishment of the Argentine Olympic Committee. This essay explores the reception and diffusion of Olympism in Argentina. It reveals a process of gradual adoption including conflicting views on the relationship between the state and sport, several attempts at institutionalization, international misunderstandings and the role of politics and class. PMID- 18286741 TI - The FTC contact lens rule: a common sense approach to compliance. PMID- 18286740 TI - Changing minds, saving lives: Franz Kafka as a key industrial reformer. PMID- 18286743 TI - "Shooting from the hip" rarely works. PMID- 18286742 TI - eHealth Blueprint outlines health information technology implementation in the United States. PMID- 18286744 TI - Use of the National Provider Identifier in ordering and referring. PMID- 18286745 TI - Absence of prolyliminopeptidase-negative Neisseria gonorroeae strains in Ontario, Canada. PMID- 18286746 TI - Effect of high salt concentrations on water structure. AB - The characteristic tetrahedral structure of water is known to be disrupted by changes in pressure and temperature. It has been suggested that ions in solution may have a similar perturbing effect. Here we use neutron diffraction to compare the effects of applied pressure and high salt concentrations on the hydrogen bonded network of water. We find that the ions induce a change in structure equivalent to the application of high pressures, and that the size of the effect is ion-specific. Ionic concentrations of a few moles per litre have equivalent pressures that can exceed a thousand atmospheres. We propose that these changes may be understood in terms of the partial molar volume of the ions, relative to those of water molecules. The equivalent induced pressure of a particular ion species is correlated with its efficacy in precipitating, or salting-out, proteins from solution. PMID- 18286747 TI - [An eastern city in the West: ethnic culinary pioneers in The Hague]. PMID- 18286748 TI - From holism to context: recent anthropological analyses of Northern Ireland. PMID- 18286749 TI - ACHE recognizes four, while Harborview wins McGaw prize. PMID- 18286750 TI - Feminist ideas and domestic violence policy change. PMID- 18286751 TI - [The development of a technique for female sterilization: three hallmarks of recent reproductive history in Latin America]. PMID- 18286752 TI - "Dusky damsels": Pitcairn Island's neglected matriarchs of the Bounty saga. PMID- 18286753 TI - [The history of the family and the complexity of social change. Part 1]. PMID- 18286754 TI - [The history of the family and the complexity of social change. Part 2]. PMID- 18286755 TI - Therapeutic action in modern conflict theory. AB - Recognizing that principles of psychoanalytic technique and conceptions of the analytic treatment process follow from the theory of therapeutic action to which they are linked, the author notes the difficulty of coming up with such a theory in relation to modern conflict theory. After reviewing Freud's initial descriptions of psychoanalytic theory and technique, as well as his later elaborations and modifications, the author summarizes the contributions of Freud's analytic contemporaries and traces the emergence of later theoretical variability in the field. He then presents an overview of recent developments in the theory of therapeutic action, discussing in particular the contributions of Arlow, Brenner, and Gray. PMID- 18286756 TI - On therapeutic action. AB - The author presents her views on therapeutic action in the light of an examination of key Freudian works. She also discusses the work of some psychoanalytic thinkers who followed Freud, such as Klein and Green. Lacan's thinking and his influence on French psychoanalysis are summarized. A detailed clinical vignette is presented to illustrate the author's use of the psychoanalytic method to ameliorate a patient's troubled mental functioning. PMID- 18286757 TI - On the therapeutic action of psychoanalysis. AB - The author discusses therapeutic action as addressed in contributions by Klein, the post-Kleinians, and Willy and Madeleine Baranger He highlights the roles played by psychoanalytic listening and psychoanalytic neutrality in therapeutic action, and presents a detailed clinical vignette to illustrate his points. PMID- 18286758 TI - The Kleinian theory of therapeutic action. AB - The author distinguishes three types of change seen in patients over the course of a psychoanalysis, noting that analysts are most interested in the type that occurs uniquely as a result of the analysis itself. He discusses Freud's views on the analytic relationship and contrasts these with the way the relationship is conceptualized within object relations psychoanalysis, and he compares Freudian views of transference and countertransference with Kleinian ideas. The use of interpretation is also examined from different theoretical viewpoints. The centrality of aggression in the Kleinian view of the psyche is put forth as a potentially controversial aspect of Kleinian technique. PMID- 18286759 TI - The mechanisms of cure in psychoanalysis. AB - The author considers the theoretical contributions of Lacan and Bion in relation to therapeutic action or the mechanisms of the cure. Whereas Bion felt that the analysand should ultimately experience transformation in O, Lacan described the analysand's goal as not to give in to one's desire or to be one's self. The author distinguishes among various types of neurotic and psychotic structures in discussing the limits of the cure, noting that the analyst's acts--as well as his words--can function as analytic interpretations. Lacan's theories of jouissance, the sexual phantom, identification with the analytic function, and post-analytic effects are also discussed. PMID- 18286760 TI - Therapeutic action in self psychology. AB - The author summarizes Kohut's principal theories and their implications for understanding therapeutic action. He notes that Kohut's model of self development can be applied to both healthy and pathological outcomes, and that this model necessitates modifications in classical psychoanalytic technique. A discussion of some of the many variations within self psychology includes elaborations of Kohut's beliefs that have been contributed by more recent theorists. The author also discusses the centrality of affects in the formation of psychic structure and the implications for technique of this theoretical construct. PMID- 18286761 TI - Intersubjectivity, therapeutic action, and analytic technique. AB - The author defines the therapeutic action of psychoanalysis as the patient's increased capacity to make changes in his/ her attitudes or behaviors in order to achieve greater wellbeing and satisfaction in life. Although most analytic theories generally agree about this, the author notes, they diverge in their specifications of the principles of analytic technique that will best accomplish this aim. The patient's experience of benefit is the most accurate criterion for evaluating the success of the analysis and thus of the resultant therapeutic action, in the author's belief An extended clinical vignette is presented in which he illustrates how his technical decisions are guided by these principles. PMID- 18286762 TI - A home for the mind. AB - The author presents his view that the patient must find a home in the analyst's mind within which to tolerate the work of analysis. Analytic work and change are facilitated by the patient's experience of the analyst's mind as a place within which the patient exists as an internal object, toward whom the analyst relates with agency and freedom. To illustrate his way of working with the patient to accomplish this, the author presents case vignettes from his own practice and from the writing of Mitchell (1997, 2000) and Steiner (1994). PMID- 18286763 TI - The concept of therapeutic action today: lights and shadows of pluralism. PMID- 18286764 TI - Comments on therapeutic action. PMID- 18286765 TI - Who needs theory of therapeutic action? PMID- 18286766 TI - Pity the poor pluralist. PMID- 18286767 TI - Therapeutic action: convergence without consensus. PMID- 18286768 TI - The therapeutic action of psychoanalysis: controversies and challenges. PMID- 18286769 TI - The theory of therapeutic action. PMID- 18286770 TI - In search of a theory of therapeutic action. PMID- 18286771 TI - Sri Lanka clinical trials registry. PMID- 18286772 TI - The role of anorectal physiology studies in the investigation of anorectal and anal sphincter disorders. PMID- 18286773 TI - Management and outcome of residual bile duct stones following cholecystectomy at a tertiary referral centre in Sri Lanka. AB - BACKGROUND: In spite of the advent of pre- and per-operative imaging techniques, the problem of residual bile duct stones following laparoscopic or open cholecystectomy still exists with a reported frequency of 2-10%. Endoscopic stone extraction is a minimally invasive technique which is fast becoming popular in the management of residual ductal stones. OBJECTIVE: To review the experience in our unit with regard to clinical presentation and the outcome following endoscopic management of residual bile duct stones. DESIGN: Retrospective descriptive study. SETTING: Patients referred to a tertiary referral centre with suspected residual bile duct stones following open or laparoscopic cholecystectomy from 5 March 2002 to 31 December 2006. PATIENTS AND METHOD: 56 patients with suspected residual bile duct stones were reviewed with regard to clinical presentation, stone profile, success rate and outcome. RESULTS: Mean age of the sample was 43.2 years. Female to male ratio was 34/22. 4 (73.3%). Main presenting symptom was epigastric or right hypochondrial pain (39.2%). Presence of residual stones or gravel noted in 91% (51) during ERCP. Complete stone extraction was achieved in 83.9% (47). All these patients became completely asymptomatic. Four patients had ERCP related morbidity. There were no deaths. Longest follow up was 38 months and the shortest 1.5 months. CONCLUSIONS: Endoscopic approach is a safe and effective method in the diagnosis and treatment of residual bile duct stones. PMID- 18286774 TI - The pH of commercially available rinse-off products in Sri Lanka and their effect on skin pH. AB - INTRODUCTION: Initially linked to antimicrobial function, the acidic skin pH plays a key role in permeability barrier homeostasis and integrity of the stratum corneum. Barrier recovery is delayed when acutely perturbed skin sites are exposed to a neutral pH. OBJECTIVE: To evaluate the pH of commercially available rinse-off products in Sri Lanka, and the effect of detergent rinses on skin pH and its recovery rate. METHODS: The pH of 18 rinse-off products was determined using pH indicator paper and a pH meter. The effect of an alkaline (pH 9) and an acid (pH 5.5) rinse-off product on the hand skin pH was compared in 48 healthy volunteers after single and multiple applications. The skin pH of the dorsum of hands was measured in nurses before (n = 131) and during (n = 40) a duty shift that involved frequent hand washing using alkaline soap. RESULTS: Soaps available in Sri Lanka have a pH of 9.1-10.5. The pH of syndets and cleansers range from 5.5-7.0. Five minutes after hand washing, the mean skin pH increased by 1.7 +/- SD 0.5 pH units with alkaline soap, and by 0.8 +/- SD 0.4 pH units with acidic cleanser (p < 0.0001). Recovery of pH was slower when alkaline soap was used. The increase in skin pH was significantly greater when hands were repetitively washed with alkaline soap (p < 0.0001). The mean skin pH values of nurses before (4.9 +/ SD 0.4) and during (5.7 +/- SD 0.7) the work shift were significantly different (p < 0.0001). CONCLUSIONS: Alkalinisation with rinse-off products increases the skin pH with potential functional and clinical implications. PMID- 18286775 TI - Use of ambulatory oesophageal pH monitoring to diagnose gastrooesophageal reflux disease. AB - OBJECTIVES: To investigate the characteristics of patients referred for ambulatory oesophageal pHmetry, the referral pattern, and the role of oesophageal pHmetry as a diagnostic tool in a tertiary care hospital in Sri Lanka. DESIGN: Data obtained from patients referred to the Gastrointestinal Research Laboratory at Teaching Hospital Ragama for 24-hour ambulatory oesophageal pHmetry from 1998 to 2005 were reviewed. Patients' upper gastrointestinal endoscopy reports and stationary oesophageal manometry reports were also reviewed. RESULTS: 140 pHmetry studies were performed during the study period. The majority of patients (> 90%) were referred by specialists in gastroenterology practicing in hospitals in or around Colombo. There were 88 males and the median age was 37 years (range 0.5 74). The median duration of symptoms was 4.1 years (range 0.2 -25). Typical symptoms of gastro-oesophageal reflux disease (GORD) were present in 53 (38%) patients, atypical symptoms of GORD in 31 (22%), and nonspecific upper gastrointestinal symptoms in 56 (40%). Pathological acid reflux was found in 43 (31%) patients. Of them, 29 (67%) had typical GORD symptoms, 7 (16%) had atypical symptoms, and 7 (16%) had non-specific upper gastrointestinal symptoms. No significant association was found between endoscopy and pHmetry results. CONCLUSION: Oesophageal pH monitoring helped to establish a cause for the morbidity in a significant number of patients with GORD symptoms, but not in the majority of patients referred for the test. In our experience the investigation does not seem to be optimally used as a diagnostic tool. PMID- 18286776 TI - Hormone receptor expression and Her/2neu amplification in breast carcinoma in a cohort of Sri Lankans. AB - OBJECTIVE: 1) To determine the profile of estrogen and, progesterone receptors (ER, PR) expression and Her/2neu amplification in carcinoma of breast of Sri Lankan women. 2) To determine their inter-relationships, and associations with age at diagnosis and histological grade. DESIGN: Retrospective analysis of data. SETTING: Department of Pathology, Faculty of Medicine, University of Peradeniya. MATERIALS: 124 samples of invasive ductal carcinoma of breast, stained for steroid receptors and Her/2neu amplification with immunohistochemistry. MEASUREMENTS: 1) Semiquantative scores of steroid receptors and Her/2neu amplification. 2) Correlations of ER, PR, Her/2neu amplification, age at diagnosis and histological grade. RESULTS: The prevalence of ER, PR and Her/2neu amplification were 53.2%, 50% and 14.6% respectively. The expression of ER and PR were significantly correlated (p < 0.001). and had a significant negative correlation with Her/2neu amplification (p0.003 each). Lower grade of the tumour was significantly related to the expression of ER (p0.000) and PR (p0.000) but not to Her/2neu amplification (p0.331). Age at diagnosis was significantly correlated to the expression of ER (p0.004), but not to PR (p0.365) or Her/2neu amplification (p0.456). CONCLUSION: Prevalence or ER, PR and Her/2neu amplification in carcinoma of breast among Sri Lankans is similar to that described internationally. The correlations of ER, PR, Her/2neu amplification, to each other, age at diagnosis and grade of tumour is as reported in other countries. PMID- 18286777 TI - Refractory wheezing: paradoxical vocal cord motion. PMID- 18286778 TI - Hyper IgM syndrome in an infant. PMID- 18286779 TI - Inflammatory pseudotumour of the liver caused by a migrated fish bone. PMID- 18286780 TI - Blue rubber bleb naevus syndrome. PMID- 18286781 TI - Hypocalcaemic fits not responding to intravenous calcium therapy. PMID- 18286782 TI - Surgical emphysema and pneumomediastinum following mastoidectomy. AB - Surgical emphysema and pneumomediastinum are rare complications of mastoidectomy probably resulting from the use of a compressed air-powered mastoid drill. Early diagnosis and appropriate management according to the severity is vital. PMID- 18286783 TI - Picture quiz 3. Left ventricular failure; mitral regurgitation secondary to papillary muscle dysfunction. PMID- 18286784 TI - Outcome of skin sensitivity testing for predicting reactions to rabies equine immunoglobulin. PMID- 18286785 TI - Strongyloides stercoralis isolated by agar plate culture. PMID- 18286786 TI - Where are the geriatric nurses? Multidisciplinary forums need your expertise. PMID- 18286787 TI - Assessing heart failure in long-term care facilities. PMID- 18286788 TI - Analysis of an integrated clinical decision support system in nursing home clinical information systems. AB - This descriptive study evaluated a clinical decision support system (CDSS) in three nursing homes. The CDSS included alerts for decline in condition, improvement in condition, constipation, dehydration, loss of skin integrity, weight loss, and weight gain. Frequencies of alerts and their triggers were counted, and Spearman's rank correlations were determined between active alert frequency and number of secondary diagnoses. The most frequent alerts were for dehydration and improvement in condition. One significant positive correlation existed between number of secondary diagnoses and weight gain alert frequencies in residents with cerebral vascular accident. Health care leaders face challenges constructing an implementation strategy that addresses usability, ensures accuracy, and guides users to critical information about residents. PMID- 18286789 TI - Older Oklahomans' knowledge, attitudes, and behaviors related to advance directives. AB - This study sought to document older Oklahomans' knowledge, attitudes, and behaviors regarding the decision to write an advance directive. In Oklahoma, 67% of individuals older than age 18 do not have an advance directive; 76% say the reason is that they "have not gotten around to it." The findings from this study also indicate that some groups in Oklahoma, because of their age and race, need nurses to initiate this conversation. The discussion about advance directives should be part of the health promotion teaching provided by nurses and presented to the general public as a normal part of health care, with the focus on planning for a comfortable and peaceful death. PMID- 18286790 TI - Normal pressure hydrocephalus: are you missing the signs? AB - Although normal pressure hydrocephalus (NPH) is a potentially treatable neurological condition, many individuals with NPH will never receive treatment because the diagnosis is missed and the symptoms are attributed to other disorders. Left untreated, NPH causes significant physical and emotional suffering and eventually leads to premature death. It is imperative that patients with gait problems, slowness of thought or actions, or urinary incontinence be evaluated and screened for NPH. Geriatric nurses are in the best position to intervene by recognizing the signs of NPH and making early and appropriate referrals. PMID- 18286791 TI - Hearing impairment: significant but underassessed in primary care settings. AB - In this study, 91 older adults with hearing impairment were interviewed and asked whether their primary care providers had ever inquired about or initiated screening for hearing loss. A total of 85% indicated there had been no physician inquiry, and additional data demonstrated that the rate of inquiry was unrelated to either objective or subjective levels of hearing loss. In addition, interview narratives revealed that hearing impairment was sometimes dismissed by primary care providers, with detrimental consequences for both the individuals with hearing impairment and their communication partners. Nurses could positively influence the initiation of treatment for hearing loss by incorporating screening techniques into their assessment routines. PMID- 18286792 TI - Family members' responsibilities to nursing home residents: "she is the only mother I got". AB - Findings from this qualitative study indicate that family members of nursing home residents hold themselves responsible for overseeing the care of their loved one, representing the resident's perspective and history, and keeping the family connections. These role expectations can be assets to nursing homes. Nursing and social work staff are called on to be leaders among all staff to maximize constructive family involvement and minimize the stress families may experience if they are not able to fulfill their role expectations. PMID- 18286793 TI - You are never too old: one woman's journey to self-actualization. PMID- 18286794 TI - Relationships: the basic building blocks of life. PMID- 18286795 TI - Fistula management. PMID- 18286796 TI - Historical and cultural aspects of man's relationship with addictive drugs. AB - Our taste for addictive psychoactive substances is attested to in the earliest human records. Historically, psychoactive substances have been used by (i) priests in religious ceremonies (eg, amanita muscaria); (ii) healers for medicinal purposes (eg, opium); or (iii) the general population in a socially approved way (eg, alcohol, nicotine, and caffeine). Our forebears refined more potent compounds and devised faster routes of administration, which contributed to abuse. Pathological use was described as early as classical Antiquity. The issue of loss of control of the substance, heralding today's concept of addiction, was already being discussed in the 17th century. The complex etiology of addiction is reflected in the frequent pendulum swings between opposing attitudes on issues that are still currently being debated, such as: is addiction a sin or a disease; should treatment be moral or medical; is addiction caused by the substance; the individual's vulnerability and psychology, or social factors; should substances be regulated or freely available. PMID- 18286797 TI - Opioids, dopamine, stress, and the addictions. AB - The articulated goals of Dialogues in Clinical Neuroscience are to serve as "an interface between clinical neuropsychiatry and the neurosciences by providing state-of-the-art information and original insights into relevant clinical, biological, and therapeutic aspects." My laboratory the Laboratory of the Biology of Addictive Diseases at The Rockefeller University, has for years been focused on "bidirectional translational research," that is, learning by careful observations and study in patient populations with the disorders under study, in this case primarily specific addictive diseases, and then using that knowledge to create improved animal models or other laboratory-based research paradigms, while, at the same time, taking research findings made at the bench into the clinic as promptly as that is appropriate and feasible. In this invited review, therefore, the focus will be on perspectives of our Laboratory of the Biology of Addictive Diseases and related National Institutes of Health/National Institute on Drug Abuse research Center, including laboratory-based molecular neurobiological research, research using several animal models designed to mimic human patterns of drug abuse and addiction, as well as basic clinical research, intertwined with treatment-related research. PMID- 18286798 TI - Neural basis of reward and craving--a homeostatic point of view. AB - Here, it is argued that the interoceptive system, which provides information about the subject's internal state and is integrated in the insular cortex, and not the subcortical ventral striatum, is the critical neural substrate for reward related processes. Understanding the internal state of the individual, which is processed via this system, makes it possible to develop new interventions that are aimed at treating reward-dysfunction disorders, ie, substance and alcohol dependence. Although the ventral striatum is important for signaling the degree to which rewarding stimuli are predicted to occur, this system alone cannot account for the complex affective, cognitive, and behavioral phenomena that occur when individuals come into contact with potentially rewarding stimuli. On the other hand, the interoceptive system is able to make connections between all cortical, subcortical, and limbic systems to orchestrate a complex set of responses. Craving and urges are among the most notable responses, and may have important functions to preserve homeostasis. PMID- 18286799 TI - Cocaine and amphetamine-like psychostimulants: neurocircuitry and glutamate neuroplasticity. AB - Although the pharmacology of amphetamine-like psychostimulants at dopamine transporters is well understood, addiction to this class of drugs has proven difficult to deal with. The reason for this disconnection is that while the molecular mechanism of amphetamine action is critical to reinforce drug use, it is only the first step in a sequence of widespread neuroplastic events in brain circuitry. This review outlines the affect of psychostimulants on mesocorticolimbic dopamine projections that mediate their reinforcing effect, and how this action ultimately leads to enduring pathological neuroplasticity in glutamatergic projections from the prefrontal cortex to the nucleus accumbens. Molecular neuroadaptations induced by psychostimulant abuse are described in glutamate neurotransmission, and from this information potential pharmacotherapeutic targets are identified, based upon reversing or countermanding psychostimulant-induced neuroplasticity. PMID- 18286800 TI - Outpatient Long-term Intensive Therapy for Alcoholics (OLITA): a successful biopsychosocial approach to the treatment of alcoholism. AB - Alcohol dependence is a frequent, chronic, relapsing, and incurable disease with enormous societal costs. Thus, alcoholism therapy and research into its outcome are of major importance for public health. The present article will: (i) give a brief overview of the epidemiology, pathogenesis, and treatment outcomes of alcohol dependence; (ii) introduce the basic principles of outpatient long-term therapy of alcohol-dependent patients; and (iii) discuss in detail process outcome research on Outpatient Long-term Intensive Therapy for Alcoholics (OLITA). This successful biopsychosocial approach to the treatment of alcoholism shows a 9-year abstinence rate of over 50%, a re-employment rate of 60%, and a dramatic recovery from comorbid depression, anxiety disorders, and physical sequelae. The outcome data are empirically based on treatment processes that have proven high predictive validity and give concrete information about where to focus the therapeutic efforts. Thus, process-outcome research on OLITA can serve for the development of new therapeutic guidelines on adapting individual relapse prevention strategies. PMID- 18286802 TI - Therapeutic options and challenges for substances of abuse. AB - Addiction to substances continues to be a significant public health concern in the United States. The following review of current pharmacological treatments discusses a range of substances: nicotine, alcohol, cocaine, and opioids. The goal is to provide an overview of currently available and new pharmacological treatments for substance use disorders, while also addressing the pharmacotherapeutic challenges remaining. The significant advances in pharmacotherapy have had limited utilization, however. For example, naltrexone for alcoholism is infrequently prescribed, buprenorphine for opiates still has relatively few qualified prescribers, and stimulants have no Food and Drug Administration-approved pharmacotherapy. These pharmacotherapies are needed, with the rate of even the relatively uncommon abuse of opiates now rising sharply. PMID- 18286801 TI - Cannabinoids in health and disease. AB - Cannabis sativa L. preparations have been used in medicine for millenia. However, concern over the dangers of abuse led to the banning of the medicinal use of marijuana in most countries in the 1930s. Only recently, marijuana and individual natural and synthetic cannabinoid receptor agonists and antagonists, as well as chemically related compounds, whose mechanism of action is still obscure, have come back to being considered of therapeutic value. However, their use is highly restricted. Despite the mild addiction to cannabis and the possible enhancement of addiction to other substances of abuse, when combined with cannabis, the therapeutic value of cannabinoids is too high to be put aside. Numerous diseases, such as anorexia, emesis, pain, inflammation, multiple sclerosis, neurodegenerative disorders (Parkinson's disease, Huntington's disease, Tourette's syndrome, Alzheimer's disease), epilepsy, glaucoma, osteoporosis, schizophrenia, cardiovascular disorders, cancer, obesity, and metabolic syndrome related disorders, to name just a few, are being treated or have the potential to be treated by cannabinoid agonists/antagonists/cannabinoid-related compounds. In view of the very low toxicity and the generally benign side effects of this group of compounds, neglecting or denying their clinical potential is unacceptable- instead, we need to work on the development of more selective cannabinoid receptor agonists/antagonists and related compounds, as well as on novel drugs of this family with better selectivity, distribution patterns, and pharmacokinetics, and--in cases where it is impossible to separate the desired clinical action and the psychoactivity--just to monitor these side effects carefully. PMID- 18286803 TI - Pharmacogenetic aspects of addictive behaviors. AB - Addictions are illnesses of complex causation, including inheritance and a role for gene/environment interactions. Functional alleles influencing pharmacodynamic (tissue response) and pharmacokinetic (absorption, distribution, and metabolism) play a role, but these interact with diverse environmental factors including early life stress, underage drug exposure, availability of addictive agents, and response to clinical interventions including pharmacotherapies. Identification of genetic factors in addiction thus plays an important role in the understanding of processes of addiction and origins of differential vulnerabilities and treatment responses. PMID- 18286805 TI - Survey for the amphibian chytrid Batrachochytrium dendrobatidis in Hong Kong in native amphibians and in the international amphibian trade. AB - Chytridiomycosis, caused by the pathogen Batrachochytrium dendrobatidis, is responsible for many amphibian declines and has been identified in wild amphibian populations on all continents where they exist, except for Asia. In order to assess whether B. dendrobatidis is present on the native amphibians of Hong Kong, we sampled wild populations of Amolops hongkongensis, Paa exilispinosa, P. spinosa and Rana chloronota during 2005-2006. Amphibians infected with B. dendrobatidis have been found in the international trade, so we also examined the extent and nature of the amphibian trade in Hong Kong during 2005-2006, and assessed whether B. dendrobatidis was present in imported amphibians. All 274 individuals of 4 native amphibian species sampled tested negative for B. dendrobatidis, giving an upper 95% confidence limit for prevalence of 1.3%. Approximately 4.3 million amphibians of 45 species from 11 countries were imported into Hong Kong via air over 12 mo; we did not detect B. dendrobatidis on any of 137 imported amphibians sampled. As B. dendrobatidis generally occurs at greater than 5% prevalence in infected populations during favorable environmental conditions, native amphibians in Hong Kong appear free of B. dendrobatidis, and may be at severe risk of impact if it is introduced. Until it is established that the pathogen is present in Hong Kong, management strategies should focus on preventing it from being imported and decreasing the risk of it escaping into the wild amphibian populations if imported. Further research is needed to determine the status of B. dendrobatidis in Hong Kong with greater certainty. PMID- 18286804 TI - Pharmacologic treatments for opioid dependence: detoxification and maintenance options. AB - While opioid dependence has more treatment agents available than other abused drugs, none are curative. They can, however, markedly diminish withdrawal symptoms and craving, and block opioid effects due to lapses. The most effective withdrawal method is substituting and tapering methadone or buprenorphine. alpha 2 Adrenergic agents can ameliorate untreated symptoms or substitute for agonists if not available. Shortening withdrawal by precipitating it with narcotic antagonists has been studied, but the methods are plagued by safety issues or persisting symptoms. Neither the withdrawal agents nor the methods are associated with better long-term outcome, which appears mostly related to post detoxification treatment. Excluding those with short-term habits, the best outcome occurs with long-term maintenance on methadone or buprenorphine accompanied by appropriate psychosocial interventions. Those with strong external motivation may do well on the antagonist naltrexone. Currently, optimum duration of maintenance on either is unclear. Better agents are needed to impact the brain changes related to addiction. PMID- 18286806 TI - Prediction model for sequence variation in the glycoprotein gene of infectious hematopoietic necrosis virus in California, U.S.A. AB - The influence of spatio-temporal factors on genetic variation of infectious hematopoietic necrosis virus (IHNV) is an active area of research. Using host isolate pairs collected from 1966 to 2004 for 237 IHNV isolates from California and southern Oregon, we examined genetic variation of the mid-G gene of IHNV that could be quantified across times and geographic locations. Information hypothesized to influence genetic variation was environmental and/or fish host demographic factors, viz. location (inland or coastal), year of isolation, habitat (river, lake, or hatchery), the agent factors of subgroup (LI or LII) and serotype (1, 2, or 3), and the host factors of fish age (juvenile or adult), sex (male or female), and season of spawning run (spring, fall, late fall, winter). Inverse distance weighting (IDW) was performed to create isopleth maps of the genetic distances of each subgroup. IDW maps showed that more genetic divergence was predicted for isolates found inland (for both subgroups: LI and LII) than for coastal watershed isolates. A mixed-effect beta regression with a logit link function was used to seek associations between genetic distances and hypothesized explanatory factors. The model that best described genetic distance contained the factors of location, year of isolation, and the interaction between location and year. Our model suggests that genetic distance was greater for isolates collected from 1966 to 2004 at inland locations than for isolates found in coastal watersheds during the same years. The agreement between the IDW and beta regression analyses quantitatively supports our conclusion that, during this time period, more genetic variation existed within subgroup LII in inland watersheds than within coastal LI isolates. PMID- 18286807 TI - Quorum sensing signals are produced by Aeromonas salmonicida and quorum sensing inhibitors can reduce production of a potential virulence factor. AB - Many pathogens control production of virulence factors by self-produced signals in a process called quorum sensing (QS). We demonstrate that acyl homoserine lactone (AHL) signals, which enable bacteria to express certain phenotypes in relation to cell density, are produced by a wide spectrum of Aeromonas salmonicida strains. All 31 typical strains were AHL producers as were 21 of 26 atypical strains, but on a strain population basis, production of virulence factors such as protease, lipase, A-layer or pigment did not correlate with the production and accumulation of AHLs in the growth medium. Pigment production was only observed in broth under highly aerated conditions. Quorum sensing inhibitors (QSIs) are compounds that specifically block QS systems without affecting bacterial growth and 2 such compounds, sulphur-containing AHL-analogues, reduced production of protease in a typical strain of Aeromonas salmonicida. The most efficient compound N-(heptylsulfanylacetyl)-L-homoserine lactone (HepS-AHL), reduced protease production by a factor of 10. Five extracellular proteases were detected on gelatin-containing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gels and 3 of these were completely down regulated by HepS-AHL. Hence, QSIs can curb virulence in some strains and could potentially be pursued as bacterial disease control measures in aquaculture. PMID- 18286808 TI - Sortase inhibitor phenyl vinyl sulfone inhibits Renibacterium salmoninarum adherence and invasion of host cells. AB - Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmonid fishes, is a Gram-positive diplococcobacillus belonging to the family Micrococcaceae. Analysis of the genome sequence of the bacterium demonstrated the presence of a sortase homolog (srtD), a gene specifying an enzyme found in Gram positive bacteria and required for covalent anchoring of cell surface proteins. Interference of sortase activity is being examined as a target for therapeutic prevention of infection by several pathogenic Gram-positive bacterial species. In silico analysis identified 8 open reading frames containing sortase recognition motifs, suggesting these proteins are translocated to the bacterial cell wall. The sortase and potential sortase substrate genes are transcribed in R. salmoninarum, suggesting they encode functional proteins. Treatment of R. salmoninarum with phenyl vinyl sulfone (PVS) significantly reduced bacterial adherence to Chinook salmon fibronectin. In addition, the ability of the PVS treated bacteria to adhere to Chinook salmon embryo cells (CHSE-214) in vitro was dramatically reduced compared to that of untreated bacteria. More importantly, PVS-treated bacteria were unable to invade and replicate within CHSE-214 cells (demonstrated by an intracellular growth assay and by light microscopy). When treated with PVS, R. salmoninarum was not cytopathic to CHSE-214 cells, whereas untreated bacteria produced cytopathology within a few days. These findings clearly show that PVS, a small molecule drug and a known sortase inhibitor, can interfere with the ability of R. salmoninarum to adhere and colonize fish cells, with a corresponding decrease in virulence. PMID- 18286809 TI - Infection by a black spot-causing species of Uvulifer and associated opercular alterations in fishes from a high-desert stream in Wyoming. AB - Black spot is a common disease syndrome of freshwater fishes. This study provides information on the rank of density of the black spot agent and opercular bone alterations associated with at least one digenean, Uvulifer sp., infecting native and non-native catostomids and cyprinids of the Upper Colorado River Basin. We evaluated the density rank of pigmented metacercariae and associated alterations in the operculum of the bluehead sucker Catostomus discobolus, flannelmouth sucker C. latipinnis, white sucker C. commersoni, catostomid hybrids, roundtail chub Gila robusta, and creek chub Semotilus atromaculatus, sampled from Muddy Creek, Wyoming, USA in 2003 or 2004. All fish species contained individuals that exhibited gross signs of the black spot agent. Bluehead and flannelmouth suckers had 100% prevalence of infection. Although the other suckers and chubs contained encysted metacercariae in at least one individual, the presence of pigmented metacercariae was not apparent (i.e. based on gross observations) in many individuals. Catostomids had higher densities of metacercariae than cyprinids, as shown by frequency distributions of density ranks. Opercular holes (i.e. holes that completely penetrated the opercle and were in direct association with the pigment associated metacercariae) and pockets (depressions on the external surface of the opercle associated with metacercariae) were abundant among catostomids but rare among cyprinids. PMID- 18286810 TI - Distribution and abundance of the salmonid parasite Parvicapsula minibicornis (Myxozoa) in the Klamath River basin (Oregon-California, U.S.A.). AB - The distribution and abundance of the myxosporean parasite Parvicapsula minibicornis in the Klamath River mirrored that of Ceratomyxa shasta, with which it shares both its vertebrate and invertebrate host. Assay of fish held at sentinel sites and water samples collected from those sites showed that parasite prevalence was highest below Iron Gate dam, which is the barrier to anadromous salmon passage. Above this barrier parasite levels fluctuated, with the parasite detected in the free-flowing river reaches between reservoirs. This was consistent with infection prevalence in the polychaete host, Manayunkia speciosa, which was greater than 1% only in populations tested below Iron Gate dam. Although a low prevalence of infection was detected in juvenile out-migrant fish in the Trinity River, the tributaries tested did not appear to be a significant source of the parasite to the mainstem despite the presence of large numbers of infected adult salmon that migrate and spawn there. Rainbow trout became infected during sentinel exposure, which expands the host range for P. minibicornis and suggests that wild rainbow trout populations are a reservoir for infection, especially above Iron Gate dam. High parasite prevalence in the lower Klamath River is likely a combined effect of high spore input from heavily infected, spawned adult salmon and the proximity to dense populations of polychaetes. PMID- 18286811 TI - Description of Myxobolus gayerae sp. n. and re-description of M. leuciscini infecting European chub from the Hungarian stretch of the river Danube. AB - Myxobolus gayerae sp. n. and M. leuciscini Gonzalez-Lanza & Alvarez-Pellitero, 1985 (Myxozoa: Myxobolidae) have been described and re-described from European chub Leuciscus cephalus L. from the Hungarian stretch of the river Danube. The ellipsoidal plasmodia of M. gayerae sp. n. were found in the mucosa of the intestinal wall, whereas the large, elongated plasmodia of M. leuciscini infected the afferent arteries of the gill filaments. The spores of M. gayerae sp. n. are relatively large, slightly oval and almost rectangular in shape. On the basis of spore morphology and 18S rDNA sequences, the most similar species was M. cycloides Gurley, 1893, but the 2 species differed in host and tissue tropism as well as in the size of the spores. The spores of M. leuciscini from L. cephalus, having no intercapsular appendix or occasionally a very small one, showed a high morphological similarity to spores collected from L. cephalus cabeda, Chondrostoma polylepis and Rutilus arcasi in Spain and described as M. leuciscini Gonzalez-Lanza & Alvarez-Pellitero, 1985. PMID- 18286812 TI - In vitro culture technique for Cryptocaryon irritans, a parasitic ciliate of marine teleosts. AB - A medium for the in vitro culture of Cryptocaryon irritans, which is an obligatorily parasitic ciliate of marine teleosts and causes 'white spot disease', was developed. The medium consisted of a layer of cultured fish cells (FHM), with an agarose gel layer covering the cell layer. The agarose gel contained 0.22% agarose, 10% fetal calf serum, 100 I.U. ml(-1) Penicillin G potassium and 100 microg ml(-1) streptomycin sulphate. Theronts of C. irritans transformed to trophonts and grew to 180 microm in mean length in the medium, although they gradually decreased in number. When trophonts fully developed in medium were transferred into seawater 4 d after inoculation, approximately 70% of them transformed to encysted tomonts and released theronts. When fish were challenged with theronts obtained from in vitro-raised parasites, approximately 40% of the theronts were recovered from fish, indicating comparative infectivity of in vitro-raised theronts to those of in vivo-raised theronts. This is the first report that C. irritans fully developed in vitro and its entire life cycle was completed without a host fish. PMID- 18286813 TI - Chemoreception in the salmon louse Lepeophtheirus salmonis: an electrophysiology approach. AB - The search for effective and long-term solutions to the problems caused by salmon lice Lepeophtheirus salmonis (Kroyer, 1837) has increasingly included biological/ecological mechanisms to combat infestation. One aspect of this work focuses on the host-associated stimuli that parasites use to locate and discriminate a compatible host. In this study we used electrophysiological recordings made directly from the antennule of adult lice to investigate the chemosensitivity of L. salmonis to putative chemical attractants from fish flesh, prepared by soaking whole fish tissue in seawater. There was a clear physiological response to whole fish extract (WFX) with threshold sensitivity at a dilution of 10 . When WFX was size fractionated, L. salmonis showed the greatest responses to the water-soluble fractions containing compounds between 1 and 10 kDa. The results suggest that the low molecular weight, water-soluble compounds found in salmon flesh may be important in salmon lice host choice. PMID- 18286814 TI - Diel variation in egg-laying by the freshwater fish louse Argulus foliaceus (Crustacea: Branchiura). AB - Removal of deposited eggs could be a useful control strategy for the damaging fish ectoparasite Argulus foliaceus, but focused control requires knowledge of egg-laying patterns. Here, we investigated diel changes in the egg-laying behaviour of a natural population of A. foliaceus. Data were collected from 17-28 May 2004. Days were divided into 3 time periods: 06:00-14:00, 14:00-22:00 and 22:00-06:00 h. Significantly more egg clutches were laid from 06:00-14:00 h than during the other 2 time periods, which were not significantly different from each other. Significantly more egg clutches per hour were laid during hours of daylight as compared to hours of darkness. Significantly more egg clutches were laid in the top 1 m of the water column than at the bottom, and this was consistent throughout all 3 time periods. It is suggested that the increase in egg-laying activity during daylight hours may be due to a higher motivation to search for hosts during the night and an increased ability to locate silhouetted egg-laying sites during the day. These data can provide information useful for egg removal and control strategies. PMID- 18286815 TI - Dorsoventral transposition of the heart chambers in sturgeon Acipenser naccarii alevins. AB - This is the first description of a dorsoventral transposition of the heart chambers in sturgeons Acipenser naccarii. The affected individuals were 2 farmed alevins aged 9 and 10 d posthatching, respectively. One was examined by light microscopy and the other by scanning electron microscopy. In both cases, the atrium and sinus venosus occupied a left ventrolateral position, the ventricle, conus arteriosus and bulbus arteriosus were located dorsally, and the transverse septum was incomplete. The anomalous heart examined by light microscopy did not differ histologically from normal hearts of similar developmental stages. The abnormal dorsoventral arrangement of the heart chambers was presumably due to a distortion of the morphogenetic movements that bring the ventricle to the ventral and the atrium to the dorsal position. The present findings, together with genetic data reported in the literature, suggest that the defective cardiac phenotype detected in the present specimens might result from a mutation affecting the sturgeon ortholog of the zebrafish overlooped (olp) gene. PMID- 18286816 TI - The t(4;8) is mediated by homologous recombination between olfactory receptor gene clusters, but other 4p16 translocations occur at random. AB - The t(4;8)(p16;p23) is the second most common constitutional chromosomal translocation and is caused by an ectopic meiotic recombination between the olfactory receptor gene clusters (ORGC), located on chromosome 4p and 8p. Given that ORGCs are scattered across the genome and make-up about 0.1% of the human genome we reasoned that translocations between 4p16 and other chromosomes might be mediated by ectopic recombination between different ORGC. In 13 patients, we mapped the breakpoints of either a balanced or unbalanced translocation between chromosome 4p16 and different chromosomes. For all four t(4;8) cases, the breakpoints fall within the 4p and 8pter ORGC, confirming that non-allelic homologous recombination (NAHR) between the ORGC is the main mechanism of the t(4;8) formation. For the nine other translocations, the breakpoints on chromosome 4 mapped to different loci, one of them within the ORGC and in two flanking the ORGC. In these three cases, the translocation breakpoint at the reciprocal chromosome did not contain ORGC sequences. We conclude that only the t(4;8) is mediated by NAHR between ORGC. PMID- 18286817 TI - A theoretical psychological perspective on predictive testing for late onset hereditary diseases. AB - Because of the burden of serious late onset hereditary diseases, psychological counselling is commonly included in the predictive testing procedure. We discuss the components of psychological counselling in the context of neurodegenerative disorders such as Huntington's disease, some hereditary cancers and some hereditary heart diseases. Psychological counselling should be tailored to the specific characteristics of the disease: penetrance and expression of the mutation, variability of the age at onset and efficacy of prevention and treatment, if available. Finally theories that have already been used for the psychological framing of counselling in the context of predictive testing or to formulate research hypotheses (e.g. Transactional Model of Stress and Coping, Common Sense Model of Self-regulation) are discussed. PMID- 18286818 TI - Paracentric inversion of Yq and review of the literature. AB - We report on the second prenatal diagnosis of familial paracentric inversion of the long arm of Y chromosome [46, X, inv(Y)(q11.2q12)]. The anomaly was detected through an amniocentesis performed because of advanced maternal age. The inversion has been detected by standard GTG banding methods and better characterized by FISH with painting probe and specific satellite probes DYZ1 and DYZ3. The inversion derived from phenotypically normal father. Pregnancy was uneventful and an healthy child was born. We discuss the issue concerning genetic prenatal counselling of this rare condition and we report the clinical follow up of the child. PMID- 18286819 TI - Frequency of the 35delG allele causing nonsyndromic recessive deafness in the Algerian patients. AB - Deafness is a heterogeneous disorder showing different patterns of inheritance and involving a multitude of different genes. Mutations in the GJB2 gene encoding connexin 26 (Cx26) protein are a major cause for non-syndromic autosomal recessive and sporadic deafness. Among these mutations, the c.35delG deletion is the most common mutation for sensorineural deafness. One hundred sixteen persons from fifty-eight families were tested by the method based on the principle of PCR mediated-site-directed mutagenesis (PSDM), followed by a Bsl1 digestion. Mutation c.35delG was diagnosed in sixteen families (11 homozygotes and 5 heterozygotes). The low allelic frequency (17.24%) and low ratio of individuals homozygous (13.8%) and heterozygous (6.9%) for the c.35delG mutation suggest that there are other mutations in the GJB2 gene or other genes responsible for deafness in the Algerian population. This study reports a significant association (P=0.003) between first cousin consanguinity and non-syndromic prelingual deafness. PMID- 18286820 TI - A boy with small supernumerary marker chromosome X identified by FISH. AB - Marker or ring X chromosomes are frequently seen in Ullrich-Turner Syndrome with 46,X,r(X) karyotype, but only 8 children were reported with an extra marker X chromosome in at least some of their cell lines, we describe a 5 years old male patient who is mosaic (17%) for a cell line with an extra ring shaped marker X chromosome in addition to a normal 46,XY cell line. He had mild motor mental retardation, a dysmorphic face, dysplastic ears, high arched palate, cryptorchidism and brachydactyly. G-banding showed 46,XY[83]/47,XY,+r?[17] karyotype. NOR banding revealed no satellite region but its centromere was intact in C-banding. By fluorescent in situ hybridization (FISH) technique, dual X/Y alpha-satellite probes were used to detect the origin of ring shaped marker chromosome and 17% of his cells had two X chromosome signals due to marker X; hybridization with X chromosome inactivation center (XIST) specific probe revealed the absence of the locus on the ring chromosome. In this report, clinical features of our patient are compared with previously reported cases and the cytogenetic and molecular cytogenetic techniques used to detect origin of marker chromosome are discussed. PMID- 18286821 TI - Supernumerary chromosome der(22)t(11;22): Emanuel syndrome associates with novel features. AB - Emanuel syndrome results from +der(22)t(11q23;22q11). Cleft palate, ear anomalies, heart defects, genital anomalies, hypotonia, and mental retardation are the main features of the syndrome. We report a nine-year-old boy with the t(11;22)(q23;q11) chromosome, transmitted in an unbalanced fashion from his mother, and originated in the maternal grandmother's meiosis. In addition to mental retardation, hypotonia, craniofacial anomalies, and cryptorchidism, he has novel findings such as, joint hyperextensibility, left liver lobe agenesis, left sided malposition of the gallbladder and pancreas hypoplasia. This is the first report associating these features with Emanuel syndrome. PMID- 18286822 TI - Partial trisomy 1(q25qter) due to a de novo unbalanced 1;19 translocation in a neonate. AB - We describe a male neonate with a duplication of 1(q25qter) due to de novo unbalanced translocation (1;19)(q25;pter). He had macrocephaly, wide sutures and wide anterior fontanelle, bilateral temporoparietal bossing, downward slanting palpebral fissures, low set, posteriorly rotated ears, downturned mouth corners, thin upper lip, retrognathia, high arched palate, triangular face, widely spaced nipples, bilateral single transverse palmar creases, bilateral partial syndactyly between second and third toes, ventricular dilatation, corpus callosum hypoplasia, and cavum septi pellucidi and cavum vergae, ventricular and atrial septal defects. PMID- 18286823 TI - A Y/15 translocation in a 45,X male with Prader-Willi syndrome. AB - We report a male neonate with a 45 X karyotype; the long arm of a chromosome 15 was translocated onto the proximal long arm of the Y chromosome. Breakpoints were identified by in situ fluorescence hybridization (FISH) on the proximal 15q13 and Yq11.2. The derivative chromosome has no primary centromere. Clinical features were compatible with Prader-Willi syndrome. This is the first report case ofmonosomy 15q and Yq deletion with Prader-Willi syndrome. PMID- 18286824 TI - Phenotypic variability in Micro syndrome: report of new cases. AB - The authors describe seven Egyptian patients (5 males and two females) with microcephaly, mild microphthalmia, microcornea, congenital cataracts and hypogenitalism (only in males). These features (after excluding possible non genetic causes) are consistent with the diagnosis of Micro syndrome. Clinical, neurological, ophthalmologic examinations and brain imaging and electrophysiological studies were performed in all patients. Three cases had characteristic facial features consistent with those originally described in the Micro syndrome whilst the rest of the cases had clearly different facies to that of the original patients of Micro syndrome but similar to those described in Martsolf syndrome. The patients had a variable degree of brain atrophy but hypogenesis of the corpus callosum was evident only in five patients. Abnormal gyral pattern, small cerebellum, vermian hypoplasia and delayed myelination were additional imaging findings in 3 cases. All patients had delayed visual evoked potential but normal electroretinogram. The frequently-reported parental consanguinity emphasizes the major role of the single gene inheritance. Mutation analysis for two patients showed homozygous nonsense mutation of RAB3GAP1 in one while the other showed no evidence of linkage to either RAB3GAP1 or RAB2GAP2. Based on these cases and review of the literature, RAB3GAP genes dysregulation may result in a spectrum of phenotypes that range from Micro syndrome to Martsolf syndrome. PMID- 18286825 TI - Transposition of great arteries in an infant born after prenatal diagnosis of trisomy 20 mosaicism. AB - We report a case of prenatally diagnosed mosaic trisomy 20 in cells cultured from amniotic fluid. Trisomy 20 was present in 7 cells (13 percent) in a total of 52 investigated cells. Following the normal findings of an ultrasound scan, the couple decided to continue the pregnancy. A dysmorphic infant was born at the 38 weeks of gestation with generalized dysmorphic features and multiple cardiac anomalies including transposition of great arteries. Chromosome analysis on both cord blood and placenta at birth revealed a normal 46,XX karyotype. This patient is the first case of a liveborn infant with mosaic trisomy 20 cells detected in amniotic fluid culture with transposition of great arteries, atrioventricular concordance and ventricoarterial discordance. PMID- 18286826 TI - Ethnic specific background of mutations in Bulgarian patients with Wilson disease. PMID- 18286827 TI - Lethal multiple pterygium syndrome associated with omphalocele. PMID- 18286828 TI - Genetic counseling of prenatally detected unbalanced t(Y;15)(q12;p13). PMID- 18286829 TI - Improving brain recovery after craniotomy. PMID- 18286830 TI - Recovery and neurological evaluation. AB - Recovery from general anaesthesia is a period of intense stress for patients: there is sympathetic activation, catecholamine release, and increase in blood pressure or heart rate. Stressful events increase cerebral blood flow and cerebral oxygen consumption, potentially producing elevation of intracranial pressure and thus, favouring cerebral insults. Measures to prevent agitation, hypertension, shivering, and coughing are therefore very well justified in neurosurgical patients. The rationale for a "rapid-awakening-strategy" after craniotomy with general anaesthesia is that an early diagnosis of postoperative neurological complications is essential to limit potentially devastating consequences and finally improve patient outcome. A trial of early recovery may always be attempted to perform a neurological evaluation. An awake patient is the best and the cheapest neuromonitoring available. If, after surgery, a patient does not rapidly recover consciousness, or a focal neurological deficit becomes apparent, a head CT-scan should be performed as soon as possible to rule out a neurosurgical complication. Close monitoring during the first 24 hours after craniotomy is mandatory. PMID- 18286831 TI - Postoperative management of adult central neurosurgical patients: systemic and neuro-monitoring. AB - Postoperative neurosurgical patients are at risk of developing complications. Systemic and neuro-monitoring are used to identify patients who deteriorate in order to treat the underlying cause and minimize the impact on outcome. Hypotension and hypoxia are likely to be the most frequent insults and can be detected easily with blood pressure monitoring and pulse oximetry. Repeated clinical examination, however, is probably the most important monitor in the postoperative setting. Clinical scores such as the Glasgow Coma Score and the more recently introduced FOUR Score are important tools to standardize the clinical assessment. Intracranial pressure monitoring, cerebral blood flow monitoring, electroencephalography, and brain imaging are often used postoperatively. Despite the numerous publications on this topic only few studies address the impact of postoperative monitoring on outcome. Accordingly, in most patients the decision on which monitors are to be used must be based on the patient's presentation and clinical judgment. PMID- 18286832 TI - Respiratory care. AB - PURPOSE OF THE REVIEW: Neurosurgical patients frequently develop respiratory complications, adversely affecting neurologic outcome and survival. The review summarizes current literature and management of respiratory complications associated with brain injury. MAJOR FINDINGS: Respiratory complications are commonly associated with traumatic brain injury and subarachnoid haemorrhage. Lung-protective ventilation with reduced tidal volumes improves outcome in acute lung injury, and should be applied to neurosurgical patients in the absence of increased intracranial pressure. Weaning from the mechanical ventilation should be initiated as soon as possible, although the role of neurological status in the weaning process is not clear. Prevention of pneumonia and aspiration improves survival. In patients with difficult weaning, early bedside percutaneous tracheostomy should be considered. FURTHER INVESTIGATIONS: Further studies are warranted to elucidate an optimal oxygenation and ventilation in brain-injured patients, weaning strategies, predictors of the failed weaning and extubation, respiratory support in patients with difficulties to wean, and early tracheostomy. PMID- 18286833 TI - Cardiovascular therapy of neurosurgical patients. AB - The causes of postoperative cardiovascular disturbances in neurosurgical patients include direct cardiac neurogenic effects, clinical situations where brain tissue is underperfused, and hyperdynamic states. EKG and echographic abnormalities are common in subarachnoid hemorrhage where cardiac troponin I is the most useful predictor of cardiac risk after SAH. Neurogenic pulmonary edema is short lived and often resolves with resolution of the neurologic problem. In traumatic brain injury, where areas of ischemia co-exist with luxury perfusion, advanced hemodynamic monitoring and prevention of jugular venous desaturation best avoid secondary brain injury and achieve optimal neurologic outcome. Induced hypertension improves blood flow through vessels compromised by cerebral stenting, angioplasty, microcatheters, thrombolysis, carotid clamping, intracranial bypass and cerebral vasospasm. Hyperdynamic lesions include vascular breakthrough after elimination of cerebral arteriovenous malformations, but also emergence hypertension and hyperemia. Pharmacologic agents and adjunctive measures are effective in controlling both the systemic and the cerebral circulation. PMID- 18286834 TI - Volume and electrolyte management. AB - Osmolality is the primary determinant of water movement across the intact blood brain barrier (BBB), and we can predict that reducing serum osmolality would increase cerebral oedema and intracranial pressure. Brain injury affects the integrity of the BBB to varying degrees. With a complete breakdown of the BBB, there will be no osmotic/oncotic gradient, and water accumulates (brain oedema) consequentially to the pathological process. In regions with very moderate BBB injury, the oncotic gradient may be effective. Finally, osmotherapy is effective in brain areas with normal BBB; hypertonic solutions (mannitol, hypertonic saline) dehydrate normal brain tissue, with a decrease in cerebral volume and intracranial pressure. In patients with brain pathology, volume depletion and/or hypotension greatly increase morbidity and mortality. In addition to management of intravascular volume, fluid therapy must often be modified for water and electrolyte (mainly sodium) disturbances. These are common in patients with neurological disease and need to be adequately treated. PMID- 18286835 TI - Prevention and treatment of intracranial hypertension. AB - Intracranial pressure (ICP) is the pressure exerted by cranial contents on the dural envelope. It comprises the partial pressures of brain, blood and cerebrospinal fluid (CSF). Normal intracranial pressure is somewhere below 10 mmHg; it may increase as a result of traumatic brain injury, stroke, neoplasm, Reye's syndrome, hepatic coma, or other pathologies. When ICP increases above 20 mmHg it may damage neurons and jeopardize cerebral perfusion. If such a condition persists, treatment is indicated. Control of ICP requires measurement, which can only be performed invasively. Standard techniques include direct ventricular manometry or measurement in the parenchyma with electronic or fiberoptic devices. Displaying the time course of pressure (high-resolution ICP tonoscopy) allows assessment of the validity of the signal and identification of specific pathological findings, such as A-, B- and C-waves. When ICP is pathologically elevated--at or above 20-25 mmHg--it needs to be lowered. A range of treatment modalities is available and should be applied with consideration of the underlying cause. When intracranial hypertension is caused by hematoma, contusion, tumor, hygroma, hydrocephalus or pneumatocephalus, surgical treatment is indicated. In the absence of a surgically treatable condition, ICP may be controlled by correcting the patient's position, temperature, ventilation or hemodynamics. If intracranial hypertension persists, drainage of CSF via external drainage is most effective. Other first-tier options include induced hypocapnea (hyperventilation; paCO2 < 35 mmHg), hyperosmolar therapy (mannitol, hypertonic saline) and induced arterial hypertension (CPP concept). When autoregulation of cerebral blood flow is compromised, hyperoncotic treatment aimed at reducing vasogenic edema and intracranial blood volume may be applied. When intracranial hypertension persists, second-tier treatments may be indicated. These include 'forced hyperventilation' (paCO2 < 25 mmHg), barbiturate coma or experimental protocols such as tris buffer, indomethacin or induced hypothermia. The last resort is emergent bilateral decompressive craniectomy; once taken into consideration, it should be performed without undue delay. PMID- 18286836 TI - Prevention and treatment of homeostatic disorders after central neurosurgical procedures. AB - Prevention and treatment of homeostatic disorders after central neurosurgical procedures requires a coordinated effort by the entire medical team caring for the patient. The goal of management is to optimize physiologic and metabolic variables so that patient outcome is improved. This chapter reviews current knowledge and clinical approaches to prevention and treatment of general homeostatic disorders that commonly complicate the postoperative course of neurosurgical patients after general anesthesia. Practice recommendations based on current clinical trials and experience will be made on the following topics: therapeutic approaches to optimal hemoglobin, cerebral blood flow and hemorrheology; prophylaxis and treatment of thrombosis; temperature management including rewarming and the control of shivering; use and side effects of anticonvulsants and corticosteroids; and therapeutic approaches to optimal serum glucose levels. PMID- 18286837 TI - Pain management after craniotomy. AB - Fear of the side effects of analgesic drugs frequently leads to the under treatment of post-craniotomy pain. Nevertheless, this pain continues to be commonly observed, is frequently severe, and, if unrelieved, may cause distress for the neurosurgical patient and serious complications for the operative brain. We review recent evidence-based data on pain therapy after intracranial surgery. Especially when performed at the end of surgery, local anaesthetic scalp infiltration provides adequate, short-term postoperative pain relief. Opioids, such as morphine or oxycodone, may be used in the early period after craniotomy. If titrated properly, opioids do not increase serious side effects as compared with codeine. The non-narcotics ketoprofen, tramadol, and paracetamol may be useful as supplemental, opioid-sparing drugs. There is a need for larger trials to delineate safety and efficacy of analgesic therapies with a focus on short- and long-term outcomes. PMID- 18286838 TI - Prevention and control of postoperative nausea and vomiting in post-craniotomy patients. AB - Postoperative nausea and vomiting (PONV) are the most frequent side-effects in the postoperative period, impairing subjective well-being and having economic impact due to delayed discharge. However, emetic symptoms can also cause major medical complications, and post-craniotomy patients may be at an increased risk. A review and critical appraisal of the existing literature on PONV in post craniotomy patients, and a comparison of these findings with the current knowledge on PONV in the general surgical population, leads to the following conclusions: (1) Despite the lack of a documented case of harm caused by retching or vomiting in a post-craniotomy patient, the potential risk caused by arterial hypertension and high intra-abdominal/intra-thoracic pressure leading to high intracranial pressure, forces to avoid PONV in these patients. (2) There is unclarity about a specifically increased (or decreased) risk for PONV in post craniotomy patients compared with other surgical procedures. (3) The decision whether or not to administer an antiemetic should not be based primarily on risk scores for PONV but on the likelihood for potential catastrophic consequences of PONV. If such a risk cannot be ruled out, a multimodal antiemetic approach should be considered regardless of the individual risk. (4) Randomized controlled trials with antiemetics in post-craniotomy patients are limited with respect to sample size and methodological quality. This also impacts upon the meaning of meta analyses performed with trials that showed marked heterogeneity and inconclusive results. (5) No studies on the treatment of established PONV are available. This highlights the need to transfer knowledge about PONV treatment from other surgical procedures. (6) Despite the possibility that PONV in post-craniotomy patients can be triggered by specific conditions (e.g. surgery near the area postrema at the floor of the fourth ventricle with the vomiting centre located nearby), recommendations based on trials in post-craniotomy patients may be flawed. Thus, general knowledge on prevention and treatment of PONV must adopted for craniotomy settings. PMID- 18286839 TI - Nurse liability considerations in hospital acquired infection (HAI). PMID- 18286840 TI - HIV case finding in settings providing care for women: a key role for nursing in the HIV epidemic. PMID- 18286841 TI - Reducing urinary tract infection: a nurse-initiated program. PMID- 18286842 TI - Act 169: Pennsylvania's new advance directive law: implications for health care providers. PMID- 18286843 TI - Strategies to prevent and control Clostridium difficile: one hospital's account. PMID- 18286844 TI - Drive-through flu clinic meets high expectations. PMID- 18286845 TI - Catheter associated blood stream infections. PMID- 18286846 TI - Hospital acquired infections: HAI. PMID- 18286847 TI - Building a strong foundation for Pennsylvania nursing. PMID- 18286848 TI - Management of comorbid insomnia. AB - Many patients with primary medical or psychiatric disorders may be at risk for the development of comorbid insomnia. Research now suggests that effective treatment and management of the comorbid insomnia may be as important as the primary disorder--especially when the relationship between the two disorders is complex. Advanced practice nurses, in particular nurse practitioners and even professional nurses are well positioned by the nature of their educational preparation to assess for sleep disorders. Insomnia is an insidious illness. It deserves to be properly treated and every individual and patient, no matter their health state, deserves a good night's rest. PMID- 18286850 TI - Massachusetts health reform forges ahead. PMID- 18286849 TI - Senators propose new framework for health system. PMID- 18286851 TI - Resilience in disaster's wake. PMID- 18286852 TI - Electing a reformer in chief. PMID- 18286853 TI - A universal lesson from Europe. PMID- 18286854 TI - Medical homes: the prescription to save primary care? PMID- 18286855 TI - Biking for better health. PMID- 18286856 TI - A surgeon opens up, and not with a scalpel. Interview by Alex Berenson. PMID- 18286857 TI - Nursing research: what is it and how can dermatology nurses use it? AB - The International Council of Nurses (2007) defines nursing research as a systemic inquiry that seeks to obtain new nursing knowledge for the benefit of patients, families, and communities and includes all aspects of health that are of interest to nursing including health promotion, disease prevention, and care during illness and recovery or towards a peaceful death. There is an increased amount of clinical research available to nurses and now we are challenged to utilize this knowledge to improve patient care. Just like Florence Nightingale, nurses today need to make observations, collect data, and use the analysis of that data to support and guide current nursing care. PMID- 18286858 TI - Literature review of theory-based empirical studies examining adolescent tanning practices. AB - Lifetime exposure to ultraviolet radiation is a major risk factor for all types of skin cancer. The purpose of this manuscript is to examine theory-guided empirical studies examining adolescent tanning practices. PMID- 18286859 TI - Herbal remedies for psoriasis: what are our patients taking? AB - The objective of this study was to review and explore the top 15 currently used and the historically used herbal remedies in the treatment of psoriasis. Articles, press releases, message boards, product marketing sites, and patient education lines through the National Library of Medicine (www.pubmed.gov), National Psoriasis Foundation (www.psoriasis.org), Google (www.google. com), and Yahoo (www.yahoo.com) were reviewed. Despite widespread use of complementary and alternative medications, specifically herbals, there is limited scientific data regarding their benefits and interactions. Studies on the efficacy and side effect profiles of these remedies are needed. Additionally, both providers and patients need to be cognizant of both potential benefit distortion and adulteration of the herbal products. PMID- 18286860 TI - What's your assessment? Blue rubber bleb nevus syndrome. PMID- 18286861 TI - Pediatric skin care: guidelines for assessment, prevention, and treatment. AB - The review of the literature suggests the pediatric population is at risk for skin breakdown and therefore pressure ulcer development. The literature reveals limited information on pediatric skin care issues in comparison to the adult population. The prevention and treatment of pressure ulcers and maintenance of skin integrity in the pediatric population often is not a high priority, especially in the critically ill child. Research has demonstrated that children differ from adults in the anatomical sites of skin breakdown; however, treatment remains the same. It is important to have an understanding of the underlying physiology of ulcer formation, the factors responsible for ulcer development, and the factors that put infants and children at risk for developing pressure ulcers. Accurate assessment, documentation, prevention, and treatment are all key factors. PMID- 18286862 TI - Phytophotodermatitis. PMID- 18286863 TI - Methods to improve employee effectiveness and productivity. PMID- 18286864 TI - Staff engagement: it starts with the leader. AB - Engagement can happen if you can take care of yourself first and respect the need for recovery times. It's very hard to find an engaged staff when the leader/manager is disengaged. Engagement is contagious. As leaders model engagement and connection, the staff will follow. Unfortunately, disengagement is also contagious and can create disastrous results. PMID- 18286865 TI - Pediatric dermatology: fungal/yeast infections, viral infections, acne, and hemangiomas. PMID- 18286866 TI - Personality and depression: a commentary. PMID- 18286867 TI - Neurobiological factors linking personality traits and major depression. AB - OBJECTIVE: To examine the neurobiological basis of personality and depression. METHOD: We examined preclinical and clinical studies related to neuroanatomy, neuroendocrine, molecular, and genetic alterations in depressed patients. We considered whether common neurobiological factors might be shared between personality and depression. RESULTS: Preclinical studies provide insights into the neurobiological mechanisms underlying the pathophysiology of depression including neuroendocrine alterations in hypothalamic-pituitary-adrenal (HPA) function, neuroanatomical alterations in key brain regions, and alterations in neurotrophin and serotonergic signalling systems. Clinical studies show similar alterations in depressed patients. Evidence suggests that neuroendocrine alterations in HPA function may contribute to personality traits. Brain regions implicated in depression, including the hippocampus and the anterior cingulate cortex, might play a role in personality. Key molecules implicated in depression have been extensively studied with reference to personality traits, particularly neuroticism. To date, physiological measures (serum and positron emission tomography) provide the strongest evidence implicating brain-derived neurotrophic factor and serotonin in personality, while genetic evidence is less convincing. CONCLUSIONS: A neurobiological link exists between personality and depression; however, more work is needed to provide an understanding of the nature of this relation and to link this work with clinical studies examining the influence of personality factors on depression. PMID- 18286868 TI - Personality and depression. AB - OBJECTIVE: To examine the implications of the association between personality and depression for the understanding, assessment, and treatment of major depression. METHOD: A broad range of peer-reviewed manuscripts relevant to personality and depression was reviewed. Particular emphasis was placed on etiology, stability, diagnosis, and treatment implications. RESULTS: Personality features in depressed samples reliably differ from those of healthy samples. The associations between personality and depression are consistent with a variety of causal models; these models can best be compared through longitudinal research. Research demonstrates that attention to personality features can be useful in diagnosis and treatment. Indeed, personality information has been on the forefront of recent efforts to advance the current diagnostic classification system. Moreover, personality dimensions have shown recent promise in the prediction of differential treatment outcome. For example, neuroticism is associated with preferential response to pharmacotherapy rather than psychotherapy. CONCLUSIONS: Consideration of personality features is crucial to the understanding and management of major depression. PMID- 18286869 TI - Validation of the alcohol use disorders identification test and the drug abuse screening test in first episode psychosis. AB - OBJECTIVE: To determine the validity and reliability of the Alcohol Use Disorders Identification Test (AUDIT) and Drug Abuse Screening Test (DAST) for detecting alcohol and drug use disorders, respectively, in a population with first-episode psychosis (FEP). METHOD: Subjects with FEP completed the AUDIT and DAST and were divided into groups according to the presence or absence of a Structured Clinical Interview for DSM-IV (SCID) diagnosis of either current alcohol or drug misuse. The data were analyzed to see whether AUDIT and DAST scores were predictive of SCID diagnosis. RESULTS: Patients with alcohol-related SCID diagnoses and those with drug-related SCID diagnoses scored significantly higher on the AUDIT and DAST, respectively, than the group without the respective SCID diagnosis (P < 0.001 in both cases). The AUDIT functioned best with a problem drinking cut-off score of 10 (sensitivity, 85%; specificity, 91%). The DAST functioned best with a problem drug use cut-off score of 3 (sensitivity, 85%; specificity, 73%). The area under the receiver operating characteristic curve was 0.86 for the AUDIT and 0.83 for the DAST. CONCLUSION: The DAST and AUDIT may reliably identify FEP patients with substance abuse. PMID- 18286870 TI - Monitoring and management of metabolic risk factors in outpatients taking antipsychotic drugs: a controlled study. AB - OBJECTIVE: To evaluate the screening, monitoring, and management of metabolic risk factors and diseases in long-term antipsychotic users in relation to current practice guidelines and current standards of care as represented by a control group from an HIV clinic. METHODS: We undertook a retrospective chart review of mental health clinic outpatients taking antipsychotic drugs long-term (cases) and HIV outpatients prescribed highly active antiretroviral therapy (control subjects). RESULTS: We included 99 mental health clinic patients and 98 HIV patients in the analysis. According to information available in the outpatient clinic chart, the 10-year coronary artery disease risk was computable for 28% of the mental health clinic patients (mean risk 11.9%) and for 90% of the HIV patients (mean risk 9.5%) (chi2 = 77.0, P < 0.001). Metabolic risk factors were less frequently documented in mental health clinic charts. All HIV clinic patients were screened for hypertension and diabetes, and 90% were screened for dyslipidemia, whereas this information was missing for 30%, 39%, and 60% of mental health clinic patients, respectively (P < 0.001 for all). Disease monitoring was also more comprehensive in HIV clinic charts (for example, 100% of HIV patients were monitored for lipids, compared with 71% of mental health clinic patients; P = 0.001). CONCLUSIONS: Improved efforts are needed in the somatic care of patients with bipolar disorder and schizophrenia who are taking antipsychotics, given that they typically have moderate-to-high risk for metabolic diseases. PMID- 18286871 TI - [Symptomatology and psychosocial adaptation in adolescents with depressive disorder and comorbid disruptive behaviour disorder]. AB - OBJECTIVE: To describe symptomatology and specific psychological, social, and academic adaptation in adolescents with depressive disorder and comorbid disruptive behaviour disorder, as well as their family situation. METHOD: Using binomial logistic regressions, this study compares adolescents with depressive disorder and comorbid disruptive behaviour disorder (n=25) with adolescents with the same behaviour problems but no comorbid depressive disorder (n=99). Sex specific interaction impacts are examined. RESULTS: While both groups have several similar characteristics, youth with a dual diagnosis have more oppositional symptoms and poorer self-esteem. Analyses show no interaction impact from sex variable. CONCLUSIONS: Adolescents in both groups would benefit from similar interventions regarding disruptive behaviour disorders and some related problems, such as using psychoactive drugs, socializing with delinquent peers, and difficulty functioning in school. Adolescents with a comorbid depressive disorder need special attention, given the more significant oppositional symptomatology and the poorer self-esteem. PMID- 18286872 TI - Stability and progression of disordered gambling: lessons from longitudinal studies. AB - OBJECTIVE: Few studies have explicitly examined the stability (that is, the tendency for individuals to stay at one diagnostic level as opposed to moving to another improved or worsened level) or progression of disordered gambling; however, conventional wisdom holds that disordered gambling is intractable and escalating. The objective of this study was to examine these assumptions. METHOD: We reviewed 5 recent prospective studies of gambling behaviour among nontreatment samples for evidence related to the stability and progression of disordered gambling. RESULTS: Our review found no evidence to support the assumption that individuals cannot recover from disordered gambling (that is, the persistence assumption), no evidence to support the assumption that individuals who have more severe gambling problems are less likely to improve than individuals who have less severe gambling problems (that is, the selective-stability assumption), and no evidence to support the assumption that individuals who have some gambling problems are more likely to worsen than individuals who do not have gambling problems (that is, the progression assumption). CONCLUSION: Contrary to professional and conventional wisdom suggesting that gambling problems are always progressive and enduring, this review demonstrates instability and multidirectional courses in disordered gambling. PMID- 18286873 TI - Collaborative mental health care for the homeless: the role of psychiatry in positive housing and mental health outcomes. AB - OBJECTIVE: Factors associated with positive outcomes for homeless men referred to a shelter-based collaborative mental health care team were examined. METHOD: A chart review of 73 clients referred over 12 months was completed. Two outcome measures were examined, clinical status and housing status, 6 months after their referral to the program. RESULT: Among the referred clients, the prevalence of severe and persistent mental illness and substance use disorders was 76.5% and 48.5%, respectively. At 6 months, 24 clients (35.3%) had improved clinically, and 33 (48.5%) were housed. Logistic regression identified 2 factors associated with clinical improvement: the number of visits with a psychiatrist and treatment adherence. The same 2 factors were associated with higher odds of housing, and presence of substance use disorder was associated with lower odds of housing at 6 month follow-up. CONCLUSION: Care by a mental health specialist is positively associated with improved outcomes. Strategies to improve treatment adherence, access to mental health specialists, and innovative approaches to treatment of substance use disorders should be considered for this population. Having a psychiatrist as a member of a shelter-based collaborative care team is one possible way of addressing the complex physical and mental health needs of homeless individuals. PMID- 18286874 TI - Re: Preferential aggregation of obsessive-compulsive spectrum disorders in schizophrenia patients with OCD. PMID- 18286875 TI - Re: The social causes of psychosis in North American psychiatry: a review of a disappearing literature. PMID- 18286876 TI - Pectus excavatum and carinatum: new concepts in the correction of congenital chest wall deformities in the pediatric age group. AB - A wide range of congenital chest wall deformities have been described over the years. The spectrum of severity may range from life threatening deformities such as cordis ectopia to those which pose functional limitations as growth and maturity approach adulthood. Until recently, pectus excavatum (PE) and carinatum(PC) malformations have generally been considered as primarily cosmetic abnormalities. "Open" surgical procedures to correct PE and PC involved extensive resection of cartilage and bone to remove the bony deformity often with lasting growth restriction of the chest wall. Minimally invasive surgery has recently been reported as an effective technique in correcting PE without removing healthy chest wall structures. Nonoperative bracing has been effectively applied to PC eliminating traditional surgical methods. This report presents the experience at the Connecticut Children's Medical Center (CCMC) in the treatment of these two common deformities using new, less invasive methods with excellent long-term results. PMID- 18286877 TI - Army health care operations in Iraq. AB - Four years of warfare in the urban environment of Iraq have produced fundamental changes in the Army's health-care system. First, improved communications and air evacuation have streamlined the transport of the wounded soldierfrom the battlefield to stateside medical centers. Second, individual ballistic armor has decreased the number of U.S. troops killed while the number of wounded soldiers has increased. Third, battling an unseen enemy has produced a marked increase in acute stress disorder, post-traumatic stress disorder and traumatic brain injury. Deployment of soldiers with chronic mental health disorders such as anxiety, attention deficit disorder, and depression is problematic. The stress of long combat tours has doubled the incidence of abuse and neglect in children of deployed service members. Comparedto active-componentsoldiers, the prevalence ofmental health disorders is twice as great in soldiers of the Army Reserve and Army National Guard. Finally, the difficulty in determining friend vs. foe in Iraq results in the incarceration of thousands of Iraqis creating both medical and ethical challenges for Army physicians. PMID- 18286878 TI - Lead aVR ST-segment elevation in acute proximal aortic dissection. PMID- 18286879 TI - Cost effective management of postoperative nausea and vomiting (PONV): a multimodal therapeutic approach. PMID- 18286880 TI - Limb loss in war: historical perspective. PMID- 18286881 TI - Reflecting upon a community hospital internship program, 1963-1975. PMID- 18286882 TI - Reducing healthcare disparities: cultural diversity training for primary care physician office practice through continuing medical education. PMID- 18286884 TI - The quest for perfection. PMID- 18286883 TI - Re: "Who should care for the patients?". PMID- 18286885 TI - Wellness and prevention. Taking wellness efforts to employers' doors. PMID- 18286886 TI - Nursing. Sun may set on LPNs as care gets more complex. PMID- 18286887 TI - Other voices. AARP aims to unify 'champions' of nursing. Interview by Matthew Weinstock. PMID- 18286888 TI - Design & construction. The quake is fake, but the lessons are real. PMID- 18286889 TI - Finance. Quality of care now a bigger actor in credit. PMID- 18286890 TI - Management. New Jersey hospitals beat GM to the punch in creating a VEBA. PMID- 18286891 TI - Technology. Speech recognition software wins converts. PMID- 18286892 TI - Storyboard. How University Hospital sliced door-to-balloon time. PMID- 18286893 TI - Quality update. Engage employees to improve staff and patient satisfaction. PMID- 18286894 TI - New perspectives. The refrain may have changed--'no mission, no margin'--but the goals remain the same. PMID- 18286895 TI - Working through the frustrations of clinical integration. AB - Everybody--hospitals, physicians, payers and even government regulators--agree that if all of a patient's providers coordinate her care, the whole process would be simpler, safer and more effective. But federal laws, the cost of information technology, physician reluctance to team with hospitals and other factors keep getting in the way. Could those obstadcles to clinical integration finally be melting away? PMID- 18286896 TI - A litmus test for critical access. AB - Skeptics say the federal critical access hospital program is too expensive and that too much of the funding is spent on construction projects. But leaders of CAHs and other proponents say upgrading aging facilities is just one step in bringing new technology and improved services to their rural communities. PMID- 18286897 TI - The profit potential of hospital labs. AB - Up to 90 percent of American hospitals have some type of laboratory outreach program, performing a test here or there for affiliated doctors in a nearby clinic. But a few hospitals have found that a more aggressive outreach program, with a relatively modest investment, can turn a cost center into a reliable profit center. PMID- 18286898 TI - The aging workforce. AB - With staffing shortages intensifying, hospitals must find ways to attract and keep older workers on the job. This foldout examines 10 strategies that workforce experts say can help hospitals recruit and retain older workers, and explains specifically how four leading health systems implement those strategies. PMID- 18286899 TI - Data page: Health care proves costly for Medicare beneficiaries. PMID- 18286900 TI - On the record. PMID- 18286901 TI - Computed tomography angiography in abdominal aortic endoleaks: what is the optimal protocol? AB - OBJECTIVE: To evaluate the roles of noncontrast and delayed phases of computed tomography angiography (CTA) for optimization of the CTA protocol in endoleak detection. METHODS: CTAs of patients who underwent abdominal aortic endovascular stent-graft placement were retrospectively reviewed. CTA comprised noncontrast, arterial, and delayed phase (2 minutes postcontrast). The arterial phase was compared with a combined arterial and delayed phase and followed by the noncontrast phase to see whether additional information was obtained. RESULTS: Thirty-eight CTA studies demonstrated endoleak. In 30 studies (79%; 95%CI 64% to 89%), endoleak was detected in the arterial phase. Eight studies (21%; 95%CI 11% to 36%) demonstrated an endoleak only in the delayed phase. No additional information was obtained in the noncontrast phase when compared with a combined reading of the arterial and delayed phases (95%CI 0% to 9%). CONCLUSION: Delayed phase imaging is necessary for endoleak detection and obviates a noncontrast phase. Identical parameters should be used for arterial and delayed phases. PMID- 18286902 TI - Implementation of a new undergraduate radiology curriculum: experience at the University of British Columbia. AB - OBJECTIVES: The purpose of this study was to review and revise the undergraduate radiology curriculum at the University of British Columbia to improve radiology education to medical students and to meet the needs of a medical program with province-wide distribution. METHODS: We identified the radiology content of the curriculum from the Curriculum Management and Information Tool online database, from personal interviews with curriculum heads, and from published information. Undergraduates' and recent graduates' opinions were solicited by means of surveys. Information on radiology curricula at medical schools across Canada was gathered from email surveys and personal contacts with members of the Canadian Heads of Academic Radiology (CHAR). RESULTS: Review of our curriculum indicated that lack of a unified syllabus resulted in redundant content, gaps in knowledge, and lack of continuity in the curriculum. Results from the survey of programs across Canada indicated that most schools also lacked a formal radiology curriculum for medical students. By adapting the guidelines from the Association of Medical Student Education in Radiology, we revised our undergraduate radiology curriculum to emphasize integration and self-learning. The modified curriculum includes a combination of instructional technology, focused lectures in preclinical years, and in-context seminars in clerkship rotations. CONCLUSION: Most medical schools in Canada do not have a formal radiology curriculum for medical students. A structured curriculum is required to improve the quality of radiology teaching for medical students. PMID- 18286903 TI - The adult tarsal navicular: why it matters. AB - The adult tarsal navicular is an integral part of the midfoot. Many unique pathological conditions affect its function, and while subtle in their presentation, they can be significant sources of pain and deformity. Recognition of these abnormalities in imaging is essential to obtaining prompt diagnoses and instigating early management interventions. In this pictorial essay, accessory ossicles, stress fractures, avascular necrosis, and tarsal coalitions involving the navicular will be discussed in terms of their characteristic imaging findings. Normal anatomy and biomechanical functional aspects of the navicular will be reviewed and multiple images from plain film, ultrasound, bone scan, computed tomography, and magnetic resonance imaging will be presented. Improved familiarity with the adult tarsal navicular and its various abnormalities will enable radiologists to recognize its importance in the differential diagnosis of midfoot pathology. PMID- 18286904 TI - Detection of suspicious malignant cervical lymph nodes of unknown origin: diagnostic accuracy of ultrasound-guided fine-needle aspiration biopsy with nodal size and central necrosis correlate. AB - OBJECTIVE: To assess the role of ultrasound-guided fine-needle aspiration biopsy (US-guided FNAB) in the diagnostic workup of suspicious malignant cervical lymph nodes in patients with palpable neck masses and without known primary cancer. The diagnostic accuracy of imaging morphologic criteria, including sizes and central necrosis for assessing suspicious malignant nodes, were also examined. METHODS: This is a retrospective study of 426 patients with palpable neck masses from an outpatient department evaluated with computed tomography, magnetic resonance imaging, or US. US-guided FNABs were performed in 102 patients with suspicious malignant cervical lymph nodes at a single institution. Cytologically positive lymph nodes (n = 12) were further validated with excisional biopsy. Negative lymph nodes (n = 90) were either excised (n = 10) or followed up by imaging studies for at least one year (n = 80). The diagnostic accuracy of the FNABs along with the imaging findings of nodal sizes and presence of central necrosis, which were classified by a consensus of 2 radiologists, were assessed. RESULTS: Twelve malignant nodes were detected with US-guided FANB with one false-positive and one false-negative result. The overall sensitivity, specificity, and accuracy for FNAB were 91.7%, 98.9%, and 98.0%, respectively. The sensitivity, specificity, and accuracy were 66.7%, 30.0%, and 34.3% for size criterion and 75.0%, 83.3%, and 82.3% for central necrosis criterion. CONCLUSIONS: The size of cervical lymph node does not appear to be an important imaging criterion for assessing suspicious malignant lymph nodes, compared with the criterion of central necrosis. US-guided FNAB is highly specific and sensitive in the diagnostic workup of suspicious malignant cervical lymph nodes in patients without known primary cancers. PMID- 18286905 TI - Answer to case of the month #124. Metallosis. Metal synovitis (periprosthetic metallosis). PMID- 18286906 TI - Answer to case of the month #125. Multiple biliary hamartomas (von Meyenburg complexes). PMID- 18286907 TI - Answer to case of the month #126. Pelvic lipomatosis in association with cystitis glandularis. PMID- 18286908 TI - Smiling hydatid. PMID- 18286909 TI - Discipline decisions raise questions. PMID- 18286910 TI - On being a provincial councillor. PMID- 18286912 TI - Panel responds to urgent questions on low birth weight. PMID- 18286911 TI - Ensuring safe and efficient communication of medication prescriptions in community and ambulatory settings. PMID- 18286913 TI - RN manager tackles issue with promising results. PMID- 18286914 TI - Students making a difference. PMID- 18286915 TI - Heroes. PMID- 18286916 TI - RN shortage needs a balanced approach. PMID- 18286917 TI - The Joint Commission adds new hospital outpatient performance measures. PMID- 18286918 TI - The Joint Commission recipients of 2007 Ernest Amory Codman Awards. National health care award for performance measurement. PMID- 18286919 TI - Clarification: details of elimination of four-month track record requirements decision. PMID- 18286920 TI - Revision: note for Standard EC.4.12, Element of Performance 6. PMID- 18286921 TI - Approved: revisions to fire-safety equipment maintenance Standard EC.5.40, Element of Performance 14. PMID- 18286922 TI - Correction: home care 2007 Update 2. PMID- 18286923 TI - Correction: "Table of Changes" for critical access hospital 2007 Update 2. PMID- 18286924 TI - Assuring quality: roles of appraisers and peer reviewers. AB - Nurses seeking continuing education opportunities have a right to know that their educational choices are well advised. Within the accreditation process, appraisers and peer reviewers play a significant role in determining that standardized criteria for continuing nursing education activities, providers, and approvers are met. PMID- 18286925 TI - Incorporating the use of thromboelastographic studies when assessing bleeding patients. AB - This column discusses the use of thromboelastographic studies when assessing patients with bleeding disorders. PMID- 18286926 TI - Difficult questions in difficult situations. AB - This column discusses effective ways to handle difficult questions posed by participants or students. PMID- 18286927 TI - Shine on: achieving career satisfaction as a registered nurse. AB - This phenomenological study focuses on the experience of career satisfaction among registered nurses. Potential participants were asked, "Do you love your work as a nurse?" A random sample of eight nurses who answered yes to this question was questioned further during semistructured conversations. Conversations were recorded and transcribed. Data collected were in the form of descriptions of times during the participants' careers when they felt most professionally fulfilled. Through narrative and poetic analysis, themes of "upholding the vulnerable," "going the extra mile," and "attending to the essential ordinary" were identified. Nurse educators play an important role facilitating career satisfaction for registered nurses. Practical implications for continuing education for educators and practicing nurses are addressed. PMID- 18286928 TI - Exploring critical thinking in critical care nursing education: a pilot study. AB - BACKGROUND: Critical care nurses process vast amounts of information and require well developed critical-thinking skills to make clinical decisions. METHOD: Using a pretest posttest design, the critical-thinking skills of 31 postgraduate nurses were evaluated using the California Critical Thinking Skills Test (CCTST). RESULTS: For the total group, mean critical-thinking scores improved slightly over time. The CCTST revealed a mean pretest score of 18.5 and a mean posttest score of 19.7, both of which were higher than the established norms for the test. Overall, no significant difference was observed between pretest and posttest. However, statistical significance was observed posttest for nurses whose critical thinking scores improved (p < .000). CONCLUSION: Despite the small sample, the majority of the postgraduate nurses improved their critical-thinking skills during the course of their study. PMID- 18286929 TI - The keys to successful online continuing education programs for nurses. AB - Asynchronous online tutorials that award continuing education units without cost and provide knowledge about computers and nursing informatics were made available to registered nurses in Southern California. Four hundred seventy-three nurses enrolled; 52% (246) completed tutorials. Nonsignificant differences in the number of tutorials completed were found across characteristics of participants, meaning that nurses were similarly disposed to participate regardless of age, educational preparation, experience, practice setting, or ethnicity. They tended to overestimate their computer capabilities at the time of enrollment and abandoned the tutorials when they encountered technical problems. Nurses need live workshops teaching computer basics, Internet skills, and how to enroll in and run asynchronous programs. Marketing of online programs should be multifaceted, including live and electronic strategies. PMID- 18286930 TI - A new type of rural nurse residency. AB - The Rural Nurse Internship program is a distance education-based nurse residency designed to meet the needs of rural hospitals across the country. Nurses learn to perform the generalist role by practicing crisis assessment and management in six subnursing specialties. The collaborative yearlong residency provides preceptors, mentors, monthly seminars, and just-in-time information to novice nurses in their own hospitals using instructional technologies. Expert rural nurses teach novice employees using a standardized curriculum. Hospitals individualize the program to meet employee and hospital needs. PMID- 18286931 TI - Reconcilable differences. PMID- 18286932 TI - Reclaiming lost revenue. PMID- 18286934 TI - Superior scheduling. PMID- 18286933 TI - Scrubbing Reimbursement Rates clean. PMID- 18286935 TI - Take me to the pilot. PMID- 18286936 TI - Mississippi Medicaid and disaster preparednes. PMID- 18286937 TI - Pursuing practice excellence. PMID- 18286938 TI - Best care, shorter stays. PMID- 18286939 TI - Beginning at the end. PMID- 18286940 TI - HIE's impact on healtcare. PMID- 18286941 TI - [Image of the month. Asherman syndrome]. PMID- 18286942 TI - [Image of the month. Small intestine perforation after an unknown ingestion of a foreign body]. PMID- 18286943 TI - [Clinical case of the month. Atrial flutter with rapid ventricular response (1:1 atrioventricular conduction) caused by flecainide]. AB - We report a case of 1:1 flutter in a patient taking flecainide for atrial fibrillation. We discuss the mechanism of the arrhythmia, its treatment and the preventive attitude to be adopted. PMID- 18286944 TI - [Clinical case of the month. Pulmonary aspergillosis: chimic necrotizing aspergillosis]. AB - Pulmonary aspergillosis are uncommon diseases. The clinical expression of which are depending on the immunological status. The spectrum of pulmonary aspergillosis goes from hypersensitization to the invasive and disseminated aspergillosis. We report here the clinical history of a patient with severe COPD and receiving regular oral corticoids who had a persistent lung infiltration. The infiltration became cavitary and proved to be a chronic necrotizing pneumonia due to Aspegillus fumigatus. The outcome was favourable thank to a treatment with voriconazole. PMID- 18286945 TI - [Clinical case of the month. Histological discovery of a lung cancer in an enormous emphysematous bulla]. AB - The development of a lung cancer in young patients with emphysematous bullae is a classical, albeit rare, event. These patients can however be operated upon for the resection of a giant bulla causing respiratory distress. We report the case of a 41 year old male patient who was submitted to the resection of a giant right apical emphysematous bulla. The histopathological examination of the fragment revealed an undifferentiated lung carcinoma the diameter of which was less than one centimeter. This observation underlines the need for a systematic examination of the resected material and for a careful radiological follow-up of such patients. PMID- 18286946 TI - [Dapsone: an opportunity to spare corticoid use for Henoch-Schonlein treatment? A case report]. AB - Dapsone is a sulfone essentially used to treat leprae. We describe, here, a somewhat rare indication of dapsone: corticoid sparing in the treatment of particular forms of Henoch-Schonlein. PMID- 18286947 TI - [Gait disorders in the elderly]. AB - Falls are frequent in the elderly and are responsible for an increased morbi mortality. The elderly who has a tendency to fall is at higher risk of repeated hospitalisation and of institutionalisation. A link between cognitive and gait disorders is put forward by several recent studies. This paper is concerned with the various gait abnormalities encountered in the elderly, especially seriale gait, and the means to detect them. PMID- 18286948 TI - [How I explore...pruritus in a pregnant woman]. AB - Any pruritus occurring in pregnant women may represent a sensorial manifestation unrelated to pregnancy, but it may represent the consequence of a pregnancy specific dermatosis. This latter group encompass pruritus gravidarum with or without intrahepatic cholestasis, pemphigoid gestationis, polymorphic eruption of pregnancy, prurigo gestationis, acute folliculitis of pregnancy, impetigo herpetiformis and the progesterone auto-immune dermatitis. Fetal risk of morbidity is recognized for pruritus gravidarum with intrahepatic cholestasis, pemphigoid gestationis and impetigo herpetiformis. PMID- 18286949 TI - [Evaluation of thymopoiesis: clinical applications]. AB - In the precedent article, we have described how T-cell generation in the thymus (thymopoiesis) may be currently evaluated through quantification by PCR of T-cell receptor excision circles (TREC) generated by intrathymic random recombination of the gene segments coding for variable parts of T-cell receptor for antigen (TCR). In hematology, TREC methodology helps in a better understanding of immune reconstitution after graft of hematopoietic stem cells: first there is a proliferation of mature T cells present in the graft, then a differentiation of naive T cells. In geriatrics, the homeostasis of the peripheral T-cell repertoire is maintained through proliferation of peripheral memory T cells rather than through thymic generation of naive T cells. In addition, TREC quantification constitutes a novel major tool for deciphering the tight control of thymopoiesis by the neuroendocrine system. PMID- 18286950 TI - [Borderline personality disorder: clinic approach of a psychopathology "without limits"]. AB - The borderline personality disorder is a common psychiatric entity, which is characterized by a varied symptomatology and a significant comorbidity. It seems that its etiologic is heterogeneous, comprising psychosocial factors as much as biological and genetic. So the best approach of the borderline personality disorder would be a multidisciplinary bio-psycho-social one. Through a clinical case, we illustrate this disorder and its treatment in outpatient clinic. PMID- 18286951 TI - [Endodontic biofilms and secondary infection of total hip arthroplasty]. AB - Biofilms are well recognised in different settings. Endodontic biofilms containing bacteria responsible for bacteraemia and secondary infection of total hip arthroplasties (THA) represent a diagnostic challenge because they are often asymptomatic. Their treatment is difficult and inadequate management of such unrecognised sources of systemic infection can lead to the devastating situation of an infected arthroplasty necessitating implant removal, long term antibiotic treatment and secondary implantation of a new prosthesis. Since about ten percent of secondary infected THA can have a dental origin, this article is intended to give guidelines to all intervening care providers: general practitioner, orthopaedic surgeon and dentist. PMID- 18286952 TI - Ocular and oculomotor findings of Joubert syndrome. PMID- 18286953 TI - Estimated cost of an ROP locum tenens program. PMID- 18286954 TI - Diplopia in adults. PMID- 18286956 TI - Ectopia lentis in childhood. PMID- 18286955 TI - What's your diagnosis? Congenital pupillary-iris-lens membrane syndrome. PMID- 18286957 TI - Long-term results of lensectomy in children with ectopia lentis. AB - PURPOSE: To report the clinical features and long-term visual results in children with ectopia lentis after lensectomy. METHODS: A retrospective analysis was performed with the records of 78 eyes of 42 patients operated on and observed for at least 3 years. RESULTS: Mean age at diagnosis was 4.6 years, mean age at surgery was 5.7 years, and follow-up period was an average of 7.1 years. Of 42 patients exhibiting bilateral ectopia lentis, 6 received monocular surgery. Mean age at diagnosis was significantly higher and mean postoperative best-corrected visual acuity was significantly lower in the monocular surgery group than in the binocular surgery group (P < .05). All eyes exhibited reduced hyperopia for 5 years postoperatively, but the reduction rate of hyperopia in the monocular group was significantly lower than inthe binocular group (P < .05). No complications occurred during operations and 1 eye (1.3%) in a patient with Marfan's syndrome exhibited retinal detachment 9 years after surgery. CONCLUSIONS: Surgical treatment of ectopia lentis was safe and promising with regard to good visual outcomes after 7.1 years of follow-up. However, the possibility of amblyopia must be considered due to differences in the degree of dislocation between the two eyes. PMID- 18286958 TI - The relationship between tumor cell differentiation and age at diagnosis in retinoblastoma. AB - PURPOSE: To determine the relationship between tumor cell differentiation and age at diagnosis in retinoblastoma. METHODS: Medical records of 170 patients with retinoblastoma treated by enucleation were reviewed retrospectively. Age at diagnosis and histopathological features were analyzed. RESULTS: Well differentiated tumors presented earlier than poorly differentiated tumors. The frequency of presentation was highest in the first year of age for well differentiated tumors and in the third year of age for poorly differentiated tumors (P < .0001). Bilateral well-differentiated tumors presented earlier than bilateral poorly differentiated tumors. Similarly, unilateral well-differentiated tumors presented earlier than unilateral poorly differentiated tumors. CONCLUSIONS: Differentiated tumors present earlier than poorly differentiated tumors, irrespective of laterality in retinoblastoma. PMID- 18286959 TI - Fundus fluorescein angiographic characterization of diffuse choroidal hemangiomas. AB - PURPOSE: To evaluate the clinical usefulness of fundus fluorescein angiography (FFA) in characterizing diffuse choroidal hemangiomas in Sturge-Weber syndrome. METHODS: A retrospective case review of children attending a university hospital clinic with a diagnosis of Sturge-Weber syndrome was undertaken. The FFA findings of seven patients who had undergone angiography were reviewed. RESULTS: FFA confirmed the presence of choroidal hemangioma in each of the seven cases and delineated the extent of the choroidal lesions. No adverse effects of fluorescein injection were encountered. CONCLUSIONS: FFA is effective in diagnosing and characterizing diffuse choroidal hemangiomas, and may be performed safely in children. PMID- 18286960 TI - The effect of surgical correction of epiblepharon on astigmatism in children. AB - PURPOSE: To evaluate the characteristics of astigmatism in epiblepharon and postoperative astigmatic changes in children. METHODS: The authors undertook a retrospective chart review of 56 patients who underwent an epiblepharon operation. Preoperative astigmatism was compared with postoperative astigmatism according to age at surgery, the presence of amblyopia, and unilateral epiblepharon. RESULTS: Astigmatism decreased from a preoperative mean of 1.34 +/- 1.25 D with-the-rule astigmatism to a postoperative mean of 1.10 +/- 1.14 D with the-rule astigmatism in all patients (P = .055), and this reduction was statistically significant in the 5- to 7-year age group (P = .038), amblyopic eyes (P = .021), and affected eyes in cases of unilateral epiblepharon (P = .004). CONCLUSIONS: Epiblepharon operations were found to change astigmatism. The results obtained suggest that an epiblepharon operation should be considered in patients with amblyopia and epiblepharon prior to the general treatment of amblyopia. PMID- 18286961 TI - Role of intravenous fluorescein angiography in early detection and regression of retinopathy of prematurity. AB - PURPOSE: To study the role of intravenous fluorescein angiography in early detection and regression of retinopathy of prematurity. METHODS: The Retcam 120 (Massie Research Laboratories, Dublin, CA) was used to perform intravenous fluorescein angiography in 40 eyes of 20 children diagnosed as having treatable retinopathy of prematurity as per Early Treatment for Retinopathy of Prematurity Cooperative Group guidelines and treated with laser photocoagulation. RESULTS: Fluorescein angiography enabled easy visualization of the retinal vasculature and helped to identify early flat neovascularization in zone 1/2 retinopathy of prematurity, detect previously missed areas of active retinopathy of prematurity in the peripheral retina, and identify skip areas following laser photocoagulation, which resulted in good management outcomes. There were no side effects related to sodium fluorescein or the procedure. CONCLUSIONS: Retcam assisted intravenous fluorescein angiography is safe and helps in the early diagnosis, prompt management, and complete regression of retinopathy of prematurity. PMID- 18286962 TI - Anterior segment ischemia after two-muscle surgery in a patient with radiation treated nasopharyngeal carcinoma. AB - Anterior segment ischemia after strabismus surgery is a rare occurrence. Risk factors include single-stage surgery of three or more muscles in one eye, dysthyroid eye disease, and advanced age. The authors report a case of anterior segment ischemia after single-stage surgery of two horizontal muscles without previous reported risk factors. However, the patient had received radiotherapy for nasopharyngeal carcinoma more than 20 years ago. The case was successfully treated with systemic steroids but with reduced best-corrected visual acuity. Previous radiotherapy in the related orbital field may be a new risk factor for this condition. PMID- 18286963 TI - Bilateral necrotizing scleritis following strabismus surgery for thyroid ophthalmopathy. AB - A case of a bilateral necrotizing scleritis following strabismus surgery for thyroid ophthalmopathy is described. A 71-year-old man was operated on for esotropia due to prior and equilibrated Grave's disease. Two weeks after surgery, he had a scleral necrosis that was located on the surgery sites (insertion of the two medial rectus muscles). There was no infectious or autoimmune etiology. The pathogenic mechanism was ischemic and inflammatory, doubtless facilitated by the thyroid ophthalmopathy. The outcome was favorable after 3 weeks of antibiotic and anti-inflammatory treatment. To the authors' knowledge, this is the first bilateral case of this complication. PMID- 18286964 TI - Pigmented free-floating vitreous cyst in a child. AB - An 8-year-old asymptomatic boy was referred for a pigmented fundus lesion in his left eye that was suspected to be a possible choroidal melanoma. Visual acuity was 20/20 in each eye. Ocular examination in the upright position revealed a pigmented lesion measuring 4 mm in diameter and 3 mm in thickness located inferior to the fovea. Following supine positioning, the mass floated to the superior macular region. UI-trasonography confirmed a cystic vitreous mass. Observation of the benign vitreous cyst was advised. A free-floating pigmented cyst can resemble a pigmented intraocular tumor, particularly if it is immediately preretinal. PMID- 18286965 TI - A rare case of retinocytoma occurring in a 12-year-old child. AB - A 12-year-old girl with a translucent, partially calcified retinal mass in the posterior pole surrounded by a rim of pigment epithelial changes and chorioretinal atrophy was diagnosed as having retinocytoma. The patient received a single treatment of transpupillary thermotherapy at the slit lamp to prevent malignant transformation. The dimensions of the lesion and the visual acuity remained unchanged at 3.5 years of follow-up with no further treatment. Retinocytoma is a benign retinal tumor that carries the same genetic implications as retinoblastoma. Regular follow-up is necessary because the tumor can demonstrate malignant transformation. PMID- 18286966 TI - Isolated orbital pseudotumor as the presenting sign of systemic lupus erythematosus. AB - A 9-year-old girl presented with an orbital pseudotumor a few years prior to developing clinical features of systemic lupus erythematosus. Because the few similar cases previously reported were all adults, this patient is the first pediatric case described. The authors also review the relevant literature, especially in the pediatric age group. PMID- 18286967 TI - Ultrasound biomicroscopy of eyelid lymphangioma in a child. AB - A 13-month-old girl developed a superficial, multinodular, blue-grey upper eyelid mass shortly after birth. This was clinically suspected to be a lymphangioma. Ultrasound biomicroscopy revealed a subcutaneous mass with a multicystic internal structure and fluid levels consistent with lymphangioma. Follow-up ultrasound biomicroscopy 2 years later showed stable findings. PMID- 18286968 TI - Abnormal head posture in a patient with normal ocular motility: Sandifer syndrome. AB - The authors describe an unusual presentation of abnormal head posture in a 5-year old boy. Sandifer syndrome is associated with torticollis and gastroesopheageal reflux. Treating physicians should be aware of non-ocular causes of abnormal head posture in the absence of ocular manifestations. PMID- 18286969 TI - Acute retinal necrosis secondary to herpes simplex virus type 2 with preexisting chorioretinal scarring. AB - Acute retinal necrosis in children is a devastating disease that requires early diagnosis and treatment. The authors describe a rarely reported case of bilateral acute retinal necrosis in a child caused by neonatal herpes simplex virus type 2, where the presence of previous chorioretinal scarring made diagnosis challenging. PMID- 18286970 TI - Posterior synechiae secondary to uveitis. PMID- 18286971 TI - Giving feedback: context is everything. A conversation with Niki Gjere, MA, MS, RN, APRN, BC. Interviewed by Marty Lewis-Hunstiger. PMID- 18286972 TI - Building resilient work environments: the role of communication. PMID- 18286973 TI - Practices of effective end-of-life communication between nurses and patients/families in two care settings. PMID- 18286974 TI - Communicating delegation--why is it so difficult? PMID- 18286975 TI - Managing defensive communication: excerpts from Taking the War out of our Words. PMID- 18286976 TI - Commitment to my Co-Workers: a brief history. PMID- 18286977 TI - Study suggests SNPs could serve disabled seniors in capitated Medicare plans. PMID- 18286978 TI - Commercial, Medicaid HMO members receive unequal care. PMID- 18286979 TI - Use benchmarks to allocate time in primary care practice. PMID- 18286980 TI - Building healthy communities through community drum circles. PMID- 18286981 TI - Clinical nurse specialists in a Web-based practice environment. AB - The clinical nurse specialists working with the Canadian Virtual Hospice (CVH) are web-based, advanced practice palliative care nursing consultants. They deal with the complexities and challenges of online communication, particularly as it relates to the sharing of information about palliative and end-of-life care with lay people and health-care professionals alike. These nurses possess a number of highly developed skills--in communication, use of evidence-based research, teaching and understanding health-care systems--that enable them to respond to questions posed by visitors to the "Ask A Professional" section of the CVH website. Although the website is administratively based in Winnipeg, Manitoba, the services of the CNSs are available to people across Canada. In this article, the authors describe how the elements of advanced practice operate within this unique delivery model. PMID- 18286982 TI - Why is it so hard to talk about spirituality? AB - Although there is recognition of the importance of spirituality in the nursing literature and in nursing theory, many nurses find it difficult to talk about this sensitive area with people for whom they provide care. In this article, the authors discuss why spirituality is integral to nursing care and explore why nurses don't talk about spiritual concerns with their clients. The authors examine the meaning of spirituality and the factors that contribute to the reluctance of nurses to discuss spirituality with others: not having the right words, lack of education, a view that spiritual care is someone else's responsibility, influences of secularism and diversity in society, and the current health-care context. Openness to learning about the spiritual beliefs of individuals and attending to their nursing needs in a holistic way will enhance nursing care. PMID- 18286983 TI - Daniel Savoie. Interview by Tara Tosh-Kennedy. PMID- 18286984 TI - Hydromorphone: handle with care. PMID- 18286985 TI - Greater integration expected in 2008. PMID- 18286986 TI - Going mobile. PMID- 18286987 TI - What's next for Medicare? PMID- 18286988 TI - Pharmacy program with DM component targets CKD. AB - Diplomat Specialty Pharmacy's CKD Navigator catches chronic kidney disease patients before they need dialysis. PMID- 18286989 TI - Program brings hypertensive patients within BP ranges. AB - A recent study focusing on hypertensive elderly patients showed telephony backed by nurses saved money. PMID- 18286990 TI - Hot topic number one: dialogal phenomenology, nursing rounds, reflective practice, and improved patient care. PMID- 18286991 TI - E-Clinic: an innovative approach to complex symptom management for allogeneic blood and stem cell transplant patients. AB - The allogeneic blood and stem cell program (ABSCP) at Princess Margaret Hospital, Toronto, performs 75 transplants annually. Many patients live greater than 100 kilometres from the centre and require frequent visits to the hospital for posttransplant care. The weekly travel to clinic, combined with complex symptom issues and the overwhelming desire to be cared for in their home community, is a major burden to patients and care providers. Our team of oncology health professionals, led by the nurse practitioner on service, sought to determine whether telehealth videoconferencing would be a viable option as a care delivery model to meet the complex needs of our remote patients and care partners. We introduced telehealth into the ambulatory clinic as a pilot project in early 2005. Patients were selected based upon symptoms, therapeutic plan and geographical remoteness. Patient progress was monitored with a goal of transitioning patients from posttransplant hospital-based care to partnered self care in their home communities. The purpose of this article is to illustrate the ABSCP telehealth program development using a patient case study, and to detail the clinical process improvements and overall program successes that have led to the integration of telehealth into everyday clinical practice as a viable service delivery option for patient-centred symptom management and treatment compliance with a geographically remote patient population. PMID- 18286992 TI - The lived experience of women with cancer: phenomenological findings expressed through poetry. AB - Cancer rates for Canadian women between the ages of 22 and 44 are increasing. Improved survival times and more treatment choices, however create new challenges. Little research has been done to uncover the lived experience of long term survival. This pilot study describes the meaning of living with cancer for three Canadian women who were diagnosed more than four years ago. The process of inquiry was Giorgi's descriptive phenomenological method for analysis-synthesis of a general structural description (the meaning of the experience). The findings have been interpreted creatively through poetry in an effort to enhance understanding of the experience of living with cancer Each section of the poem is discussed in relation to the literature to encourage nurses and other health professionals to consider the importance of understanding patients' lived experiences and the meanings they ascribe, in order to provide quality, holistic, and individualized care. PMID- 18286993 TI - Breast cancer information dissemination strategies--finding out what works. AB - This study aimed to identify the best strategies for dissemination of information about breast cancer In November 2004, 28 breast cancer survivors were interviewed. Three themes emerged from these discussions: the shock of diagnosis; the onus being on the patient to search for information; and the different types of information that breast cancer survivors want. To learn multiple viewpoints, 12 focus groups were held with breast cancer survivors (n = 127) and three focus groups were conducted with information providers (n = 25) in the spring of 2005. Participants validated the themes and identified two programs using "best practices" to provide information for women dealing with breast cancer. This article highlights the study findings, including implications for practice, education, and research. PMID- 18286994 TI - What's in a name? Student, researcher, leader, mom. Interview by Anne Remington. PMID- 18286995 TI - Hospice nursing: a final gift. AB - Hospice care remains one of the best kept secrets out there. Surveys have shown that if people could choose the circumstances for their death, they would be pain free, die at home, and be among family and friends. Yet in our country there seems to be a reluctance to allow people to let go and complete life's circle. Understandably, there is a tendency for physicians to do one more thing or try one more option--a tendency to fight the inevitable. Many health care professionals seem to feel that offering hospice is tantamount to taking away hope. They view hospice as a last resort rather than a last gift: the experience of allowing a fellow human being to have a "good death". PMID- 18286996 TI - Holistic nursing: a way of being, a way of living, a way of practice! PMID- 18286997 TI - Global nursing: as close as your own back yard or halfway 'round the world. PMID- 18286998 TI - Travel nursing 101: get the facts. PMID- 18286999 TI - Spinal cord injury nursing: an extraordinary specialty. PMID- 18287000 TI - Rehabilitation nursing: is it a fit for you? PMID- 18287001 TI - Imaging the impact on cuprate superconductivity of varying the interatomic distances within individual crystal unit cells. AB - Many theoretical models of high-temperature superconductivity focus only on the doping dependence of the CuO(2)-plane electronic structure. However, such models are manifestly insufficient to explain the strong variations in superconducting critical temperature, T(c), among cuprates that have identical hole density but are crystallographically different outside of the CuO(2) plane. A key challenge, therefore, has been to identify a predominant out-of-plane influence controlling the superconductivity, with much attention focusing on the distance d(A) between the apical oxygen and the planar copper atom. Here we report direct determination of how variations in interatomic distances within individual crystalline unit cells affect the superconducting energy-gap maximum Delta of Bi(2)Sr(2)CaCu(2)O(8+delta). In this material, quasiperiodic variations of unit cell geometry occur in the form of a bulk crystalline "supermodulation." Within each supermodulation period, we find approximately 9 +/- 1% cosinusoidal variation in local Delta that is anticorrelated with the associated d(A) variations. Furthermore, we show that phenomenological consistency would exist between these effects and the random Delta variations found near dopant atoms if the primary effect of the interstitial dopant atom is to displace the apical oxygen so as to diminish d(A) or tilt the CuO(5) pyramid. Thus, we reveal a strong, nonrandom out-of-plane effect on cuprate superconductivity at atomic scale. PMID- 18287002 TI - Retraction for Holweg and Nick: Arabidopsis myosin XI mutant is defective in organelle movement and polar auxin transport. PMID- 18287003 TI - Splicing promotes rapid and efficient mRNA export in mammalian cells. AB - The numerous steps in protein gene expression are extensively coupled to one another through complex networks of physical and functional interactions. Indeed, >25 coupled reactions, often reciprocal, have been documented among such steps as transcription, capping, splicing, and polyadenylation. Coupling is usually not essential for gene expression, but instead enhances the rate and/or efficiency of reactions and, physiologically, may serve to increase the fidelity of gene expression. Despite numerous examples of coupling in gene expression, whether splicing enhances mRNA export still remains controversial. Although splicing was originally reported to promote export in both mammalian cells and Xenopus oocytes, it was subsequently concluded that this was not the case. These newer conclusions were surprising in light of the observations that the mRNA export machinery colocalizes with splicing factors in the nucleus and that splicing promotes recruitment of the export machinery to mRNA. We therefore reexamined the relationship between splicing and mRNA export in mammalian cells by using FISH, in combination with either transfection or nuclear microinjection of plasmid DNA. Together, these analyses indicate that both the kinetics and efficiency of mRNA export are enhanced 6- to 10-fold (depending on the construct) for spliced mRNAs relative to their cDNA counterparts. We conclude that splicing promotes mRNA export in mammalian cells and that the functional coupling between splicing and mRNA export is a conserved and general feature of gene expression in higher eukaryotes. PMID- 18287004 TI - fMRI mapping of a morphed continuum of 3D shapes within inferior temporal cortex. AB - Here, we mapped fMRI responses to incrementally changing shapes along a continuous 3D morph, ranging from a head ("face") to a house ("place"). The response to each shape was mapped independently by using single-stimulus imaging, and stimulus shapes were equated for lower-level visual cues. We measured activity in 2-mm samples across human inferior temporal cortex from the fusiform face area (FFA) (apparently selective for faces) to the parahippocampal place area (PPA) (apparently selective for places), testing for (i) incremental changes in the topography of FFA and PPA (predicted by the continuous-mapping model) or (ii) little or no response to the intermediate morphed shapes (predicted by the category model). Neither result occurred; instead, we found approximately linearly graded changes in the response amplitudes to graded-shape changes, without changes in topography-similar to visual responses in different lower-tier cortical areas. PMID- 18287005 TI - Discovery of a lysophospholipid acyltransferase family essential for membrane asymmetry and diversity. AB - All organisms consist of cells that are enclosed by a cell membrane containing bipolar lipids and proteins. Glycerophospholipids are important not only as structural and functional components of cellular membrane but also as precursors of various lipid mediators. Polyunsaturated fatty acids comprising arachidonic acid or eicosapentaenoic acid are located at sn-2 position, but not at sn-1 position of glycerophospholipids in an asymmetrical manner. In addition to the asymmetry, the membrane diversity is important for membrane fluidity and curvature. To explain the asymmetrical distribution of fatty acids, the rapid turnover of sn-2 position was proposed in 1958 by Lands [Lands WE (1958) Metabolism of glycerolipides: A comparison of lecithin and triglyceride synthesis. J Biol Chem 231:883-888]. However, the molecular mechanisms and biological significance of the asymmetry remained unknown. Here, we describe a putative enzyme superfamily consisting mainly of three gene families, which catalyzes the transfer of acyl-CoAs to lysophospholipids to produce different classes of phospholipids. Among them, we characterized three important enzymes with different substrate specificities and tissue distributions; one, termed lysophosphatidylcholine acyltransferase-3 (a mammalian homologue of Drosophila nessy critical for embryogenesis), prefers arachidonoyl-CoA, and the other two enzymes incorporate oleoyl-CoAs to lysophosphatidylethanolamine and lysophosphatidylserine. Thus, we propose that the membrane diversity is produced by the concerted and overlapped reactions with multiple enzymes that recognize both the polar head group of glycerophospholipids and various acyl-CoAs. Our findings constitute a critical milestone for our understanding about how membrane diversity and asymmetry are established and their biological significance. PMID- 18287007 TI - Peptide adsorption on a hydrophobic surface results from an interplay of solvation, surface, and intrapeptide forces. AB - The hydrophobic effect, i.e., the poor solvation of nonpolar parts of molecules, plays a key role in protein folding and more generally for molecular self assembly and aggregation in aqueous media. The perturbation of the water structure accounts for many aspects of protein hydrophobicity. However, to what extent the dispersion interaction between molecular entities themselves contributes has remained unclear. This is so because in peptide folding interactions and structural changes occur on all length scales and make disentangling various contributions impossible. We address this issue both experimentally and theoretically by looking at the force necessary to peel a mildly hydrophobic single peptide molecule from a flat hydrophobic diamond surface in the presence of water. This setup avoids problems caused by bubble adsorption, cavitation, and slow equilibration that complicate the much-studied geometry with two macroscopic surfaces. Using atomic-force spectroscopy, we determine the mean desorption force of a single spider-silk peptide chain as F = 58 +/- 8 pN, which corresponds to a desorption free energy of approximately 5 k(B)T per amino acid. Our all-atomistic molecular dynamics simulation including explicit water correspondingly yields the desorption force F = 54 +/- 15 pN. This observation demonstrates that standard nonpolarizable force fields used in classical simulations are capable of resolving the fine details of the hydrophobic attraction of peptides. The analysis of the involved energetics shows that water-structure effects and dispersive interactions give contributions of comparable magnitude that largely cancel out. It follows that the correct modeling of peptide hydrophobicity must take the intimate coupling of solvation and dispersive effects into account. PMID- 18287006 TI - Gating at the selectivity filter in cyclic nucleotide-gated channels. AB - By opening and closing the permeation pathway (gating) in response to cGMP binding, cyclic nucleotide-gated (CNG) channels serve key roles in the transduction of visual and olfactory signals. Compiling evidence suggests that the activation gate in CNG channels is not located at the intracellular end of pore, as it has been established for voltage-activated potassium (K(V)) channels. Here, we show that ion permeation in CNG channels is tightly regulated at the selectivity filter. By scanning the entire selectivity filter using small cysteine reagents, like cadmium and silver, we observed a state-dependent accessibility pattern consistent with gated access at the middle of the selectivity filter, likely at the corresponding position known to regulate structural changes in KcsA channels in response to low concentrations of permeant ions. PMID- 18287008 TI - On the evolutionary significance and metal-binding characteristics of a monolobal transferrin from Ciona intestinalis. AB - Transferrins are a family of proteins that bind and transport Fe(III). Modern transferrins are typically bilobal and are believed to have evolved from an ancient gene duplication of a monolobal form. A novel monolobal transferrin, nicatransferrin (nicaTf), was identified in the primitive ascidian species Ciona intestinalis that possesses the characteristic features of the proposed ancestral Tf protein. In this work, nicaTf was expressed in Pichia pastoris. Extensive solution studies were performed on nicaTf, including UV-vis, fluorescence, CD, EPR and NMR spectroscopies, and electrospray time-of-flight mass spectrometry. The expressed protein is nonglycosylated, unlike the protein isolated from the organism. This property does not affect its ability to bind Fe(III). However, Fe(III)-bound nicaTf displays important spectral differences from other Fe(III) bound transferrins, which are likely the consequence of differences in metal coordination. Coordination differences could also account for the weaker affinity of nicaTf for Fe(III) (log K = 18.5) compared with bilobal human serum transferrin (HsTf) (log K = 22.5 and 21.4). The Fe-nicaTf complex is not labile, as indicated by slow metal removal kinetics by the high-affinity chelator tiron at pH 7.4. The protein alternatively binds up to one equivalent of Ti(IV) or V(V), which suggests that it may transport nonferric metals. These solution studies provide insight into the structure and function of the primitive monolobal transferrin of C. intestinalis for comparison with higher order bilobal transferrins. They suggest that a major advantage for the evolution of modern transferrins, dominantly of bilobal form, is stronger Fe(III) affinity because of cooperativity. PMID- 18287009 TI - Profile of Randy Schekman: reflections on his first year as PNAS editor-in-chief. PMID- 18287010 TI - Acid-sensing ion channels interact with and inhibit BK K+ channels. AB - Acid-sensing ion channels (ASICs) are neuronal non-voltage-gated cation channels that are activated when extracellular pH falls. They contribute to sensory function and nociception in the peripheral nervous system, and in the brain they contribute to synaptic plasticity and fear responses. Some of the physiologic consequences of disrupting ASIC genes in mice suggested that ASIC channels might modulate neuronal function by mechanisms in addition to their H(+)-evoked opening. Within ASIC channel's large extracellular domain, we identified sequence resembling that in scorpion toxins that inhibit K(+) channels. Therefore, we tested the hypothesis that ASIC channels might inhibit K(+) channel function by coexpressing ASIC1a and the high-conductance Ca(2+)- and voltage-activated K(+) (BK) channel. We found that ASIC1a associated with BK channels and inhibited their current. Reducing extracellular pH disrupted the association and relieved the inhibition. BK channels, in turn, altered the kinetics of ASIC1a current. In addition to BK, ASIC1a inhibited voltage-gated Kv1.3 channels. Other ASIC channels also inhibited BK, although acidosis-dependent relief of inhibition varied. These results reveal a mechanism of ion channel interaction and reciprocal regulation. Finding that a reduced pH activated ASIC1a and relieved BK inhibition suggests that extracellular protons may enhance the activity of channels with opposing effects on membrane voltage. The wide and varied expression patterns of ASICs, BK, and related K(+) channels suggest broad opportunities for this signaling system to alter neuronal function. PMID- 18287011 TI - The PD-1/PD-L1 complex resembles the antigen-binding Fv domains of antibodies and T cell receptors. AB - Signaling through the programmed death 1 (PD-1) inhibitory receptor upon binding its ligand, PD-L1, suppresses immune responses against autoantigens and tumors and plays an important role in the maintenance of peripheral immune tolerance. Release from PD-1 inhibitory signaling revives "exhausted" virus-specific T cells in chronic viral infections. Here we present the crystal structure of murine PD-1 in complex with human PD-L1. PD-1 and PD-L1 interact through the conserved front and side of their Ig variable (IgV) domains, as do the IgV domains of antibodies and T cell receptors. This places the loops at the ends of the IgV domains on the same side of the PD-1/PD-L1 complex, forming a surface that is similar to the antigen-binding surface of antibodies and T cell receptors. Mapping conserved residues allowed the identification of residues that are important in forming the PD-1/PD-L1 interface. Based on the structure, we show that some reported loss-of binding mutations involve the PD-1/PD-L1 interaction but that others compromise protein folding. The PD-1/PD-L1 interaction described here may be blocked by antibodies or by designed small-molecule drugs to lower inhibitory signaling that results in a stronger immune response. The immune receptor-like loops offer a new surface for further study and potentially the design of molecules that would affect PD-1/PD-L1 complex formation and thereby modulate the immune response. PMID- 18287012 TI - Requirement of protein kinase D1 for pathological cardiac remodeling. AB - The adult heart responds to biomechanical stress and neurohormonal signaling by hypertrophic growth, accompanied by fibrosis, diminished pump function, and activation of a fetal gene program. Class II histone deacetylases (HDACs) suppress stress-dependent remodeling of the heart via their association with the MEF2 transcription factor, an activator of heart disease. Protein kinase D (PKD) is a stress-responsive kinase that phosphorylates class II HDACs, resulting in their dissociation from MEF2 with consequent activation of MEF2 target genes. To test whether PKD1 is required for pathological cardiac remodeling in vivo, we generated mice with a conditional PKD1-null allele. Mice with cardiac-specific deletion of PKD1 were viable and showed diminished hypertrophy, fibrosis, and fetal gene activation as well as improved cardiac function in response to pressure overload or chronic adrenergic and angiotensin II signaling. We conclude that PKD1 functions as a key transducer of stress stimuli involved in pathological cardiac remodeling in vivo. PMID- 18287013 TI - MicroRNAs and the advent of vertebrate morphological complexity. AB - The causal basis of vertebrate complexity has been sought in genome duplication events (GDEs) that occurred during the emergence of vertebrates, but evidence beyond coincidence is wanting. MicroRNAs (miRNAs) have recently been identified as a viable causal factor in increasing organismal complexity through the action of these approximately 22-nt noncoding RNAs in regulating gene expression. Because miRNAs are continuously being added to animalian genomes, and, once integrated into a gene regulatory network, are strongly conserved in primary sequence and rarely secondarily lost, their evolutionary history can be accurately reconstructed. Here, using a combination of Northern analyses and genomic searches, we show that 41 miRNA families evolved at the base of Vertebrata, as they are found and/or detected in lamprey, but not in either ascidians or amphioxus (or any other nonchordate taxon). When placed into temporal context, the rate of miRNA acquisition and the extent of phenotypic evolution are anomalously high early in vertebrate history, far outstripping any other episode in chordate evolution. The genomic position of miRNA paralogues in humans, together with gene trees incorporating lamprey orthologues, indicates that although GDEs can account for an increase in the diversity of miRNA family members, which occurred before the last common ancestor of all living vertebrates, GDEs cannot account for the origin of these novel families themselves. We hypothesize that lying behind the origin of vertebrate complexity is the dramatic expansion of the noncoding RNA inventory including miRNAs, rather than an increase in the protein-encoding inventory caused by GDEs. PMID- 18287014 TI - Unfolded protein response and cell death after depletion of brefeldin A-inhibited guanine nucleotide-exchange protein GBF1. AB - Guanine nucleotide-exchange factors (GEFs) activate ADP-ribosylation factor (ARF) GTPases that recruit coat proteins to membranes to initiate transport vesicle formation. Three mammalian GEFs are inhibited by brefeldin A (BFA). GBF1, predominantly associated with cis-Golgi membranes, functions early in the secretory pathway, whereas BIG1 and BIG2 act in trans-Golgi or later sites. Perturbation of endoplasmic reticulum (ER) functions can result in accumulation of unfolded or misfolded proteins that causes ER stress and unfolded protein response (UPR), with accumulation of ER stress response element (ERSE) gene products. BFA treatment of cells causes accumulation of proteins in the ER, ER stress, and ultimately apoptosis. To assess involvement of BFA-sensitive GEFs in the damage resulting from prolonged BFA treatment, HepG2 cells were selectively depleted of BIG1, BIG2, or GBF1 by using specific siRNA. Only GBF1 siRNA dramatically slowed cell growth, led to cell-cycle arrest in G(0)/G(1) phase, and caused dispersion of Golgi markers beta-COP and GM130, whereas ER structure appeared intact. GBF1 depletion also significantly increased levels of ER proteins calreticulin and protein disulfide isomerase (PDI). Proteomic analysis identified ER chaperones involved in the UPR that were significantly increased in amounts in GBF1-depleted cells. Upon ER stress, transcription factor ATF6 translocates from the ER to Golgi, where it is sequentially cleaved by site 1 and site 2 proteases, S1P and S2P, to a 50-kDa form that activates transcription of ERSE genes. Depletion of GBF1, but not BIG1 or BIG2, induced relocation of S2P from Golgi to ER with proteolysis of ATF6 followed by up-regulation of ER chaperones, mimicking a UPR response. PMID- 18287015 TI - WAVE1 controls neuronal activity-induced mitochondrial distribution in dendritic spines. AB - Mitochondrial fission and trafficking to dendritic protrusions have been implicated in dendritic spine development. Here, we show that Wiskott-Aldrich syndrome protein (WASP)-family verprolin homologous protein 1 (WAVE1) controls depolarization-induced mitochondrial movement into dendritic spines and filopodia and regulates spine morphogenesis. Depolarization-induced degradation of the p35 regulatory subunit of cyclin-dependent kinase 5 (Cdk5), with the resultant decreased inhibitory phosphorylation on WAVE1, depend on NMDA receptor activation. Thus, WAVE1 dephosphorylation and activation are likely associated with mitochondrial redistribution and spine morphogenesis. PMID- 18287016 TI - Amino acid polymorphisms in Arabidopsis phytochrome B cause differential responses to light. AB - Plants have a sophisticated system for sensing and responding to their light environment. The light responses of populations and species native to different habitats show adaptive variation; understanding the mechanisms underlying photomorphogenic variation is therefore of significant interest. In Arabidopsis thaliana, phytochrome B (PHYB) is the dominant photoreceptor for red light and plays a major role in white light. Because PHYB has been proposed as a candidate gene for several quantitative trait loci (QTLs) affecting light response, we have investigated sequence and functional variation in Arabidopsis PHYB. We examined PHYB sequences in 33 A. thaliana individuals and in the close relative Arabidopsis lyrata. From 14 nonsynonymous polymorphisms, we chose 5 for further study based on previous QTL studies. In a larger collection of A. thaliana accessions, one of these five polymorphisms, I143L, was associated with variation in red light response. We used transgenic analysis to test this association and confirmed experimentally that natural PHYB polymorphisms cause differential plant responses to light. Furthermore, our results show that allelic variation of PHYB activity is due to amino acid rather than regulatory changes. Together with earlier studies linking variation in light sensitivity to photoreceptor genes, our work suggests that photoreceptors may be a common target of natural selection. PMID- 18287017 TI - CD4 T cells, lymphopenia, and IL-7 in a multistep pathway to autoimmunity. AB - There are many inhibitory mechanisms that function at the cellular and molecular levels to maintain tolerance. Despite this, self-reactive clones escape regulatory mechanisms and cause autoimmunity in certain circumstances. We hypothesized that the same mechanisms that permit T cells to expand during homeostatic proliferation may inadvertently promote autoimmunity under certain conditions. One major homeostatic cytokine is IL-7, and studies have linked it or its receptor to the development of multiple sclerosis and other autoimmune diseases. We show in a model of beta-islet cell self-reactivity that the transfer of activated autoreactive CD4 T cells can prime and expand endogenous autoreactive CD8 T cells in a CD28- and CD40-dependent manner through the licensing of dendritic cells. Despite this, mice do not develop diabetes. However, the provision of exogenous IL-7 or the physiological production of IL-7 associated with lymphopenia was able to profoundly promote the expansion of self reactive clones even in the presence of regulatory T cells. Autoimmune diabetes rapidly ensued with CD4 help and the subsequent activation of CD8 T cells, which contributed to disease progression. With the advent of many biologicals targeting TNFalpha, IL-6, and IL-1 and their effective use in the treatment of autoimmune diseases, we propose that IL-7 and its receptor may be promising targets for biological agents in the treatment of autoimmunity. PMID- 18287018 TI - Collagen fibril architecture, domain organization, and triple-helical conformation govern its proteolysis. AB - We describe the molecular structure of the collagen fibril and how it affects collagen proteolysis or "collagenolysis." The fibril-forming collagens are major components of all mammalian connective tissues, providing the structural and organizational framework for skin, blood vessels, bone, tendon, and other tissues. The triple helix of the collagen molecule is resistant to most proteinases, and the matrix metalloproteinases that do proteolyze collagen are affected by the architecture of collagen fibrils, which are notably more resistant to collagenolysis than lone collagen monomers. Until now, there has been no molecular explanation for this. Full or limited proteolysis of the collagen fibril is known to be a key process in normal growth, development, repair, and cell differentiation, and in cancerous tumor progression and heart disease. Peptide fragments generated by collagenolysis, and the conformation of exposed sites on the fibril as a result of limited proteolysis, regulate these processes and that of cellular attachment, but it is not known how or why. Using computational and molecular visualization methods, we found that the arrangement of collagen monomers in the fibril (its architecture) protects areas vulnerable to collagenolysis and strictly governs the process. This in turn affects the accessibility of a cell interaction site located near the cleavage region. Our observations suggest that the C-terminal telopeptide must be proteolyzed before collagenase can gain access to the cleavage site. Collagenase then binds to the substrate's "interaction domain," which facilitates the triple-helix unwinding/dissociation function of the enzyme before collagenolysis. PMID- 18287019 TI - A test of the chromosomal theory of ecotypic speciation in Anopheles gambiae. AB - The role of chromosomal inversions in speciation has long been of interest to evolutionists. Recent quantitative modeling has stimulated reconsideration of previous conceptual models for chromosomal speciation. Anopheles gambiae, the most important vector of human malaria, carries abundant chromosomal inversion polymorphism nonrandomly associated with ecotypes that mate assortatively. Here, we consider the potential role of paracentric inversions in promoting speciation in A. gambiae via "ecotypification," a term that refers to differentiation arising from local adaptation. In particular, we focus on the Bamako form, an ecotype characterized by low inversion polymorphism and fixation of an inversion, 2Rj, that is very rare or absent in all other forms of A. gambiae. The Bamako form has a restricted distribution by the upper Niger River and its tributaries that is associated with a distinctive type of larval habitat, laterite rock pools, hypothesized to be its optimal breeding site. We first present computer simulations to investigate whether the population dynamics of A. gambiae are consistent with chromosomal speciation by ecotypification. The models are parameterized using field observations on the various forms of A. gambiae that exist in Mali, West Africa. We then report on the distribution of larvae of this species collected from rock pools and more characteristic breeding sites nearby. Both the simulations and field observations support the thesis that speciation by ecotypification is occurring, or has occurred, prompting consideration of Bamako as an independent species. PMID- 18287020 TI - Quantifying changes in the thiol redox proteome upon oxidative stress in vivo. AB - Antimicrobial levels of reactive oxygen species (ROS) are produced by the mammalian host defense to kill invading bacteria and limit bacterial colonization. One main in vivo target of ROS is the thiol group of proteins. We have developed a quantitative thiol trapping technique termed OxICAT to identify physiologically important target proteins of hydrogen peroxide (H(2)O(2)) and hypochlorite (NaOCl) stress in vivo. OxICAT allows the precise quantification of oxidative thiol modifications in hundreds of different proteins in a single experiment. It also identifies the affected proteins and defines their redox sensitive cysteine(s). Using this technique, we identified a group of Escherichia coli proteins with significantly (30-90%) oxidatively modified thiol groups, which appear to be specifically sensitive to either H(2)O(2) or NaOCl stress. These results indicate that individual oxidants target distinct proteins in vivo. Conditionally essential E. coli genes encode one-third of redox-sensitive proteins, a finding that might explain the bacteriostatic effect of oxidative stress treatment. We identified a select group of redox-regulated proteins, which protect E. coli against oxidative stress conditions. These experiments illustrate that OxICAT, which can be used in a variety of different cell types and organisms, is a powerful tool to identify, quantify, and monitor oxidative thiol modifications in vivo. PMID- 18287021 TI - Dysregulation of IL-32 in myelodysplastic syndrome and chronic myelomonocytic leukemia modulates apoptosis and impairs NK function. AB - TNFalpha levels are elevated in the marrows of patients with myelodysplastic syndrome (MDS) and are associated with high rates of apoptosis, which contributes to hematopoietic failure. We observed that exposure of human marrow stroma cell lines HS5 and HS27a to TNFalpha increases levels of IL-32 mRNA. IL-32, in turn, induces TNFalpha. Marrow stroma from patients with MDS expressed 14- to 17-fold higher levels of IL-32 mRNA than healthy controls. In contrast, cells from patients with chronic myelomonocytic leukemia (CMML) expressed only one tenth the level of IL-32 measured in healthy controls. Human KG1a leukemia cells underwent apoptosis when cocultured with HS5 stromal cells, but knockdown of IL-32 in the stromal cells by using siRNA abrogated apoptosis in the leukemia cells. IL-32 knockdown cells also showed dysregulation of VEGF and other cytokines. Furthermore, CD56(+) natural killer cells from patients with MDS and CMML expressed IL-32 at lower levels than controls and exhibited reduced cytotoxic activity, which was unaffected by IL-2 treatment. We propose that IL-32 is a marrow stromal marker that distinguishes patients with MDS and CMML. Furthermore, IL-32 appears to contribute to the pathophysiology of MDS and may be a therapeutic target. PMID- 18287022 TI - The stumpy gene is required for mammalian ciliogenesis. AB - Cilia are present on nearly all cell types in mammals and perform remarkably diverse functions. However, the mechanisms underlying ciliogenesis are unclear. Here, we cloned a previously uncharacterized highly conserved gene, stumpy, located on mouse chromosome 7. Stumpy was ubiquitously expressed, and conditional loss in mouse resulted in complete penetrance of perinatal hydrocephalus (HC) and severe polycystic kidney disease (PKD). We found that cilia in stumpy mutant brain and kidney cells were absent or markedly deformed, resulting in defective flow of cerebrospinal fluid. Stumpy colocalized with ciliary basal bodies, physically interacted with gamma-tubulin, and was present along ciliary axonemes, suggesting that stumpy plays a role in ciliary axoneme extension. Therefore, stumpy is essential for ciliogenesis and may be involved in the pathogenesis of human congenital malformations such as HC and PKD. PMID- 18287023 TI - A virocidal amphipathic {alpha}-helical peptide that inhibits hepatitis C virus infection in vitro. AB - An amphipathic alpha-helical peptide (C5A) derived from the membrane anchor domain of the hepatitis C virus (HCV) NS5A protein is virocidal for HCV at submicromolar concentrations in vitro. C5A prevents de novo HCV infection and suppresses ongoing infection by inactivating both extra- and intracellular infectious particles, and it is nontoxic in vitro and in vivo at doses at least 100-fold higher than required for antiviral activity. Mutational analysis indicates that C5A's amphipathic alpha-helical structure is necessary but not sufficient for its virocidal activity, which depends on its amino acid composition but not its primary sequence or chirality. In addition to HCV, C5A inhibits infection by selected flaviviruses, paramyxoviruses, and HIV. These results suggest a model in which C5A destabilizes viral membranes based on their lipid composition, offering a unique therapeutic approach to HCV and other viral infections. PMID- 18287024 TI - Ca2+ oscillation frequency decoding in cardiac cell hypertrophy: role of calcineurin/NFAT as Ca2+ signal integrators. AB - The role of Ca(2+) signaling in triggering hypertrophy was investigated in neonatal rat cardiomyocytes in vitro. We show that an increase in cell size and sarcomere reorganization were elicited by receptor agonists such as Angiotensin II, aldosterone, and norepinephrine and by a small rise in medium KCl concentration, a treatment devoid of direct effects on receptor functions. All these treatments increased the frequency of spontaneous [Ca(2+)] transients, caused nuclear translocation of transfected NFAT(GFP), and increased the expression of a NFAT-sensitive reporter gene. There was no increase in Ca(2+) spark frequency in the whole cell or in the perinuclear region under these conditions. Hypertrophy and NFAT translocation but not the increased frequency of [Ca(2+)] transients were inhibited by the calcineurin inhibitor cyclosporine A. Hypertrophy by the different stimuli was insensitive to inhibition of myofilament contraction. We concluded that calcineurin-NFAT can act as integrators of the contractile Ca(2+) signal, and that they can decode alterations in the frequency even of rapid Ca(2+) oscillations. PMID- 18287025 TI - JNK MAP kinase activation is required for MTOC and granule polarization in NKG2D mediated NK cell cytotoxicity. AB - Interaction of the activating receptor NKG2D with its ligands is a major stimulatory pathway for cytotoxicity of natural killer (NK) cells. Here, the signaling pathway involved after NKG2D ligation is examined. Either incubation of the NKG2D-bearing human NKL tumor cell line with K562 target cells or cross linking with NKG2D mAb induced strong activation of the mitogen-activated protein (MAP) kinases. Selective inhibition of JNK MAP kinase with four different means of inhibition greatly reduced NKG2D-mediated cytotoxicity toward target cells and furthermore, blocked the movement of the microtubule organizing center (MTOC), granzyme B (a component of cytotoxic granules), and paxillin (a scaffold protein) to the immune synapse. NKG2D-induced activation of JNK kinase was also blocked by inhibitors of Src protein tyrosine kinases and phospholipase PLCgamma, upstream of JNK. Similarly, a second MAP kinase pathway through ERK was previously shown to be required for NK cell cytotoxicity. Thus, activation of two MAP kinase pathways is required for cytotoxic granule and MTOC polarization and for cytotoxicity of human NK cells when NKG2D is ligated. PMID- 18287026 TI - Allelic mutant series reveal distinct functions for Arabidopsis cycloartenol synthase 1 in cell viability and plastid biogenesis. AB - Sterols have multiple functions in all eukaryotes. In plants, sterol biosynthesis is initiated by the enzymatic conversion of 2,3-oxidosqualene to cycloartenol. This reaction is catalyzed by cycloartenol synthase 1 (CAS1), which belongs to a family of 13 2,3-oxidosqualene cyclases in Arabidopsis thaliana. To understand the full scope of sterol biological functions in plants, we characterized allelic series of cas1 mutations. Plants carrying the weak mutant allele cas1-1 were viable but developed albino inflorescence shoots because of photooxidation of plastids in stems that contained low amounts of carotenoids and chlorophylls. Consistent with the CAS1 catalyzed reaction, mutant tissues accumulated 2,3 oxidosqualene. This triterpenoid precursor did not increase at the expense of the pathway end products. Two strong mutations, cas1-2 and cas1-3, were not transmissible through the male gametes, suggesting a role for CAS1 in male gametophyte function. To validate these findings, we analyzed a conditional CRE/loxP recombination-dependent cas1-2 mutant allele. The albino phenotype of growing leaf tissues was a typical defect observed shortly after the CRE/loxP induced onset of CAS1 loss of function. In the induced cas1-2 seedlings, terminal phenotypes included arrest of meristematic activity, followed by necrotic death. Mutant tissues accumulated 2,3-oxidosqualene and contained low amounts of sterols. The vital role of sterols in membrane functioning most probably explains the requirement of CAS1 for plant cell viability. The observed impact of cas1 mutations on a chloroplastic function implies a previously unrecognized role of sterols or triterpenoid metabolites in plastid biogenesis. PMID- 18287027 TI - Antisera induced by infusions of autologous Ad-CD154-leukemia B cells identify ROR1 as an oncofetal antigen and receptor for Wnt5a. AB - We examined the sera of six patients before and after i.v. infusions of autologous chronic lymphocytic leukemia (CLL) cells transduced ex vivo with an adenovirus encoding CD154 (Ad-CD154). Five patients made high-titer antibodies against adenovirus and three made IgG reactive with a leukemia-associated surface antigen, which we identified as ROR1. Anti-ROR1 antibodies were not detected in the sera of untreated patients. We generated anti-ROR1 mAbs and found they reacted specifically with the CLL cells of all patients, but not with nonleukemic leukocytes, a wide variety of normal adult tissues, or blood mononuclear cells, including CD5(+) B cells of healthy adults. ROR1 could bind Wnt5a, which induced activation of NF-kappaB when coexpressed with ROR1 in HEK293 cells and enhanced the survival of CLL cells in vitro, an effect that could be neutralized by posttreatment anti-ROR1 antisera. We conclude that patients with CLL can break immune tolerance to ROR1, which is an oncofetal surface antigen and survival signaling receptor in this neoplastic disease. PMID- 18287028 TI - Natural selection and cultural rates of change. AB - It has been claimed that a meaningful theory of cultural evolution is not possible because human beliefs and behaviors do not follow predictable patterns. However, theoretical models of cultural transmission and observations of the development of societies suggest that patterns in cultural evolution do occur. Here, we analyze whether two sets of related cultural traits, one tested against the environment and the other not, evolve at different rates in the same populations. Using functional and symbolic design features for Polynesian canoes, we show that natural selection apparently slows the evolution of functional structures, whereas symbolic designs differentiate more rapidly. This finding indicates that cultural change, like genetic evolution, can follow theoretically derived patterns. PMID- 18287029 TI - Discovery of a selective inhibitor of oncogenic B-Raf kinase with potent antimelanoma activity. AB - BRAF(V600E) is the most frequent oncogenic protein kinase mutation known. Furthermore, inhibitors targeting "active" protein kinases have demonstrated significant utility in the therapeutic repertoire against cancer. Therefore, we pursued the development of specific kinase inhibitors targeting B-Raf, and the V600E allele in particular. By using a structure-guided discovery approach, a potent and selective inhibitor of active B-Raf has been discovered. PLX4720, a 7 azaindole derivative that inhibits B-Raf(V600E) with an IC(50) of 13 nM, defines a class of kinase inhibitor with marked selectivity in both biochemical and cellular assays. PLX4720 preferentially inhibits the active B-Raf(V600E) kinase compared with a broad spectrum of other kinases, and potent cytotoxic effects are also exclusive to cells bearing the V600E allele. Consistent with the high degree of selectivity, ERK phosphorylation is potently inhibited by PLX4720 in B Raf(V600E)-bearing tumor cell lines but not in cells lacking oncogenic B-Raf. In melanoma models, PLX4720 induces cell cycle arrest and apoptosis exclusively in B Raf(V600E)-positive cells. In B-Raf(V600E)-dependent tumor xenograft models, orally dosed PLX4720 causes significant tumor growth delays, including tumor regressions, without evidence of toxicity. The work described here represents the entire discovery process, from initial identification through structural and biological studies in animal models to a promising therapeutic for testing in cancer patients bearing B-Raf(V600E)-driven tumors. PMID- 18287030 TI - Natal dispersal driven by environmental conditions interacting across the annual cycle of a migratory songbird. AB - Natal dispersal, the process through which immature individuals permanently depart their natal area in search of new sites, is integral to the ecology and evolution of animals. Insights about the underlying causes of natal dispersal arise mainly from research on species whose short dispersal distances or restricted distributions make them relatively easy to track. However, for small migratory animals, the causes of natal dispersal remain poorly understood because individuals are nearly impossible to track by using conventional mark-recapture approaches. Using stable-hydrogen isotope ratios in feathers of American redstarts (Setophaga ruticilla) captured as immature birds and again as adults, we show that habitat use during the first tropical nonbreeding season appears to interact with latitudinal gradients in spring phenology on the temperate breeding grounds to influence the distance traveled on the initial spring migration and the direction of natal dispersal. In contrast, adult redstarts showed considerable site fidelity between breeding seasons, indicating that environmental conditions did not affect dispersal patterns after the first breeding attempt. Our findings suggest that habitat occupancy during the first nonbreeding season helps determine the latitude at which this species of Neotropical-Nearctic migratory bird breeds throughout its life and emphasize the need to understand how events throughout the annual cycle interact to shape fundamental biological processes. PMID- 18287031 TI - CNK and HYP form a discrete dimer by their SAM domains to mediate RAF kinase signaling. AB - RAF kinase functions in the mitogen-activated protein kinase (MAPK) pathway to transmit growth signals to the downstream kinases MEK and ERK. Activation of RAF catalytic activity is facilitated by a regulatory complex comprising the proteins CNK (Connector enhancer of KSR), HYP (Hyphen), and KSR (Kinase Suppressor of Ras). The sterile alpha-motif (SAM) domain found in both CNK and HYP plays an essential role in complex formation. Here, we have determined the x-ray crystal structure of the SAM domain of CNK in complex with the SAM domain of HYP. The structure reveals a single-junction SAM domain dimer of 1:1 stoichiometry in which the binding mode is a variation of polymeric SAM domain interactions. Through in vitro and in vivo mutational analyses, we show that the specific mode of dimerization revealed by the crystal structure is essential for RAF signaling and facilitates the recruitment of KSR to form the CNK/HYP/KSR regulatory complex. We present two docking-site models to account for how SAM domain dimerization might influence the formation of a higher-order CNK/HYP/KSR complex. PMID- 18287032 TI - Fine structure of the promoter-sigma region 1.2 interaction. AB - We recently proposed that a nontemplate strand base in the discriminator region of bacterial promoters, the region between the -10 element and the transcription start site, makes sequence-specific contacts to region 1.2 of the sigma subunit of Escherichia coli RNA polymerase (RNAP). Because rRNA promoters contain sequences within the discriminator region that are suboptimal for interaction with sigma1.2, these promoters have the kinetic properties required for regulation by the RNAP-binding factors DksA and ppGpp. Here, we use zero-length cross-linking and mutational, kinetic, and footprinting studies to map RNAP interactions with the nontemplate strand bases at the junction of the -10 element and the discriminator region in an unregulated rRNA promoter variant and in the lambdaP(R) promoter. Our studies indicate that nontemplate strand bases adjacent to the -10 element bind within a 9-aa interval in sigma1.2 (residues 99-107). We also demonstrate that the downstream-most base on the nontemplate strand of the 10 hexamer cross-links to sigma region 2, and not to sigma1.2. Our results refine models of RNAP-DNA interactions in the promoter complex that are crucial for regulation of transcription initiation. PMID- 18287033 TI - TRIMCyp expression in Old World primates Macaca nemestrina and Macaca fascicularis. AB - Primates have evolved a variety of restriction factors that prevent retroviral replication. One such factor, TRIM5alpha, mediates a postentry restriction in many Old World primates. Among New World primates, Aotus trivirgatus exerts a similar early restriction mediated by TRIMCyp, a TRIM5-cyclophilin A (CypA) chimera resulting from a CypA retrotransposition between exons 7 and 8 of the TRIM5 gene. Macaca nemestrina do not express TRIM5alpha; therefore, we asked whether these animals and related Old World primates express TRIMCyp. RT-PCR of total RNA from M. nemestrina and Macaca fascicularis yielded three TRIMCyp amplification products, one of which is predicted to encode a TRIMCyp chimera containing a full-length CypA. Unlike A. trivirgatus, genomic sequencing of M. nemestrina and M. fascicularis identifies a CypA retrotransposition in the 3' untranslated region of the TRIM5 locus. There is approximately 78% homology between the predicted protein sequences of Old World and New World primate TRIMCyp, with most of the differences found in the TRIM5-derived sequence. Notably, exon 7 is absent from both M. nemestrina and M. fascicularis TRIMCyp. Neither M. nemestrina nor M. fascicularis TRIMCyp could restrict HIV-1 or simian immunodeficiency virus SIVmac in an in vitro infectivity assay. The discovery of TRIMCyp in both M. nemestrina and M. fascicularis indicates that TRIMCyp expression may be more common among Old World primates than previously believed. Convergent evolution of TRIMCyp in both Old World and New World primates suggests that TRIMCyp may have provided evolutionary advantages. PMID- 18287034 TI - Independent genesis of chimeric TRIM5-cyclophilin proteins in two primate species. AB - The host range of retroviruses is influenced by antiviral proteins such as TRIM5, a restriction factor that recognizes and inactivates incoming retroviral capsids. Remarkably, in Owl monkeys (omk), a cyclophilin A (CypA) cDNA has been transposed into the TRIM5 locus, resulting in the expression of a TRIM5-CypA fusion protein (TRIMCyp) that restricts retroviral infection based on the retroviral capsid binding specificity of CypA. Here, we report that the seemingly improbable genesis of TRIMCyp has, in fact, occurred twice, and pigtailed macaques (pgt) express an independently generated TRIMCyp protein. The omkTRIMCyp and pgtTRIMCyp proteins restrict infection by several lentiviruses, but their specificities are distinguishable. Surprisingly, pgtTRIMCyp cannot bind to or restrict HIV-1 capsids as a consequence of a point mutation close to the Cyp:capsid-binding interface that was acquired during or after transposition of pgtCypA. However, the same mutation confers on pgtTRIMCyp the ability to restrict FIV in the presence of cyclosporin A, a drug that normally abolishes the interaction between pgtTRIMCyp or omkTRIMCyp and lentiviral capsids. Overall, an intuitively unlikely evolutionary event has, in fact, occurred at least twice in primates and represents a striking example of convergent evolution in divergent species. PMID- 18287035 TI - Independent evolution of an antiviral TRIMCyp in rhesus macaques. AB - The antiretroviral restriction factor TRIM5 has recently emerged as an important mediator of innate immunity and species-specific inhibition of retroviral replication in mammals. Selection pressure from pathogenic infection has driven rapid evolution of TRIM5 genes, leading to the antiviral specificities we see today. Remarkably, the New World owl monkey (Aotus trivirgatus) encodes a TRIM5 protein in which the antiviral determinants in the B30.2 domain have been replaced by cyclophilin A (CypA) encoded by a retrotransposed cDNA. The owl monkey TRIMCyp protein restricts infection by a subset of lentiviruses that recruit CypA to their capsids, including HIV-1 and feline immunodeficiency virus. Here, we show that the Old World monkey, rhesus macaque (Macaca mulatta), also encodes a TRIMCyp protein that has arisen independently from that in owl monkeys. The rhesus TRIMCyp is encoded by a single, but common, allele (Mamu7) of the rhesus TRIM5 gene, among at least six further alleles that encode full-length TRIM5 proteins with no homology to CypA. The antiviral specificity of the rhesus TRIMCyp is distinct, restricting infection of HIV-2 and feline immunodeficiency virus but not HIV-1. Restriction by rhesus TRIMCyp is before reverse transcription and inhibited by blocking CypA binding, with cyclosporine A, or by mutation of the capsid CypA binding site. These observations suggest a mechanism of restriction that is conserved between TRIMCyp proteins. The lack of activity against HIV-1 suggests that Mamu7 homozygous animals will be null for TRIM5 mediated restriction of HIV-1 and could contribute to improved animal models for HIV/AIDS. PMID- 18287036 TI - 6-Ethynylthieno[3,2-d]- and 6-ethynylthieno[2,3-d]pyrimidin-4-anilines as tunable covalent modifiers of ErbB kinases. AB - Analysis of the x-ray crystal structure of mono-substituted acetylenic thienopyrimidine 6 complexed with the ErbB family enzyme ErbB-4 revealed a covalent bond between the terminal carbon of the acetylene moiety and the sulfhydryl group of Cys-803 at the solvent interface. The identification of this covalent adduct suggested that acetylenic thienopyrimidine 6 and related analogs might also be capable of forming an analogous covalent adduct with EGFR, which has a conserved cysteine (797) near the ATP binding pocket. To test this hypothesis, we treated a truncated, catalytically competent form of EGFR (678 1020) with a structurally related propargylic amine (8). An investigation of the resulting complex by mass spectrometry revealed the formation of a covalent complex of thienopyrimidine 8 with Cys-797 of EGFR. This finding enabled us to readily assess the irreversibility of various inhibitors and also facilitated a structure-activity relationship understanding of the covalent modifying potential and biological activity of a series of acetylenic thienopyrimidine compounds with potent antitumor activity. Several ErbB family enzyme and cell potent 6-ethynyl thienopyrimidine kinase inhibitors were found to form covalent adducts with EGFR. PMID- 18287037 TI - Peptoids that mimic the structure, function, and mechanism of helical antimicrobial peptides. AB - Antimicrobial peptides (AMPs) and their mimics are emerging as promising antibiotic agents. We present a library of "ampetoids" (antimicrobial peptoid oligomers) with helical structures and biomimetic sequences, several members of which have low-micromolar antimicrobial activities, similar to cationic AMPs like pexiganan. Broad-spectrum activity against six clinically relevant BSL2 pathogens is also shown. This comprehensive structure-activity relationship study, including circular dichroism spectroscopy, minimum inhibitory concentration assays, hemolysis and mammalian cell toxicity studies, and specular x-ray reflectivity measurements shows that the in vitro activities of ampetoids are strikingly similar to those of AMPs themselves, suggesting a strong mechanistic analogy. The ampetoids' antibacterial activity, coupled with their low cytotoxicity against mammalian cells, make them a promising class of antimicrobials for biomedical applications. Peptoids are biostable, with a protease-resistant N-substituted glycine backbone, and their sequences are highly tunable, because an extensive diversity of side chains can be incorporated via facile solid-phase synthesis. Our findings add to the growing evidence that nonnatural foldamers will emerge as an important class of therapeutics. PMID- 18287038 TI - ESCRT factors restrict mycobacterial growth. AB - Nearly 1.7 billion people are infected with Mycobacterium tuberculosis. Its ability to survive intracellularly is thought to be central to its success as a pathogen, but how it does this is poorly understood. Using a Drosophila model of infection, we identify three host cell activities, Rab7, CG8743, and the ESCRT machinery, that modulate the mycobacterial phagosome. In the absence of these factors the cell no longer restricts growth of the non-pathogen Mycobacterium smegmatis. Hence, we identify factors that represent unique vulnerabilities of the host cell, because manipulation of any one of them alone is sufficient to allow a nonpathogenic mycobacterial species to proliferate. Furthermore, we demonstrate that, in mammalian cells, the ESCRT machinery plays a conserved role in restricting bacterial growth. PMID- 18287039 TI - BH3-only protein Puma contributes to death of antigen-specific T cells during shutdown of an immune response to acute viral infection. AB - During acute T cell immune responses to viral infection, antigen-specific T cells first proliferate and differentiate into effector cells, but after pathogen clearance most are deleted by apoptosis. The developmentally programmed death of antigen-specific T cells during shutdown of a T cell response is mediated by the Bcl-2-regulated apoptotic pathway and partly depends on the proapoptotic BH3-only protein Bim. However, loss of Bim enhanced survival of antigen-activated T cells to a lesser extent than Bcl-2 overexpression, indicating that other proapoptotic factors must contribute to T cell killing. In this study, we investigated the contributions of several BH3-only proteins to the shutdown of an acute T cell immune response in vivo. After infection with human herpes simplex virus (HSV-1), mice lacking Noxa, Bid, or Bad had a normal increase and subsequent decline in the numbers of antigen-specific CD8(+) T cells. In contrast, Puma-deficient mice showed an abnormally prolonged persistence of antigen-specific CD8(+) T cells in the spleen, associated with enhanced in vitro survival of these cells in the absence of cytokines. Puma was dispensable for viral clearance and also did not play a role in proliferation or activation of HSV-1-specific CD8(+) T cells in vivo. Collectively, these findings show that Puma contributes to the death of antigen-specific T cells during shutdown of an immune response. PMID- 18287040 TI - HSP90/70 chaperones are required for rapid nucleosome removal upon induction of the GAL genes of yeast. AB - Induction of transcription of the GAL genes of yeast by galactose is a multistep process: Galactose frees the activator Gal4 of its inhibitor, Gal80, allowing Gal4 to recruit proteins required to transcribe the GAL genes. Here, we show that deletion of components of either the HSP90 or the HSP70 chaperone machinery delays this induction. This delay remains when the galactose-signaling pathway is bypassed, and it cannot be explained by a chaperone requirement for DNA binding by Gal4. Removal of promoter-bound nucleosomes is delayed in a chaperone mutant, and our findings suggest an involvement of HSP90 and HSP70 in this early step in gene induction. PMID- 18287041 TI - Arabidopsis LEAFY COTYLEDON2 induces maturation traits and auxin activity: Implications for somatic embryogenesis. AB - LEAFY COTYLEDON2 (LEC2) is a central regulator of embryogenesis sufficient to induce somatic cells to form embryos when expressed ectopically. Here, we analyze the cellular processes induced by LEC2, a B3 domain transcription factor, that may underlie its ability to promote somatic embryogenesis. We show auxin responsive genes are induced after LEC2 activation in seedlings. Genes encoding enzymes involved in auxin biosynthesis, YUC2 and YUC4, are activated within 1 h after induction of LEC2 activity, and YUC4 appears to be a direct transcriptional target of LEC2. We also show ectopic LEC2 expression induces accumulation of seed storage protein and oil bodies in vegetative and reproductive organs, events that normally occur during the maturation phase of embryogenesis. Furthermore, LEC2 activates seed protein genes before an increase in RNAs encoding LEC1 or FUS3 is observed. Thus, LEC2 causes rapid changes in auxin responses and induces cellular differentiation characteristic of the maturation phase. The relevance of these changes to the ability of LEC2 to promote somatic embryogenesis is discussed. PMID- 18287042 TI - Single mutations convert an outward K+ channel into an inward K+ channel. AB - Shaker-type K(+) channels in plants display distinct voltage-sensing properties despite sharing sequence and structural similarity. For example, an Arabidopsis K(+) channel (SKOR) and a tomato K(+) channel (LKT1) share high amino acid sequence similarity and identical domain structures; however, SKOR conducts outward K(+) current and is activated by positive membrane potentials (depolarization), whereas LKT1 conducts inward current and is activated by negative membrane potentials (hyperpolarization). The structural basis for the "opposite" voltage-sensing properties of SKOR and LKT1 remains unknown. Using a screening procedure combined with random mutagenesis, we identified in the SKOR channel single amino acid mutations that converted an outward-conducting channel into an inward-conducting channel. Further domain-swapping and random mutagenesis produced similar results, suggesting functional interactions between several regions of SKOR protein that lead to specific voltage-sensing properties. Dramatic changes in rectifying properties can be caused by single amino acid mutations, providing evidence that the inward and outward channels in the Shaker family from plants may derive from the same ancestor. PMID- 18287043 TI - Acute regulation of aquaporin-2 phosphorylation at Ser-264 by vasopressin. AB - By phosphoproteome analysis, we identified a phosphorylation site, serine 264 (pS264), in the COOH terminus of the vasopressin-regulated water channel, aquaporin-2 (AQP2). In this study, we examined the regulation of AQP2 phosphorylated at serine 264 (pS264-AQP2) by vasopressin, using a phospho specific antibody (anti-pS264). Immunohistochemical analysis showed pS264-AQP2 labeling of inner medullary collecting duct (IMCD) from control mice, whereas AQP2 knockout mice showed a complete absence of labeling. In rat and mouse, pS264 AQP2 was present throughout the collecting duct system, from the connecting tubule to the terminal IMCD. Immunogold electron microscopy, combined with double labeling confocal immunofluorescence microscopy with organelle-specific markers, determined that the majority of pS264 resides in compartments associated with the plasma membrane and early endocytic pathways. In Brattleboro rats treated with [deamino-Cys-1, d-Arg-8]vasopressin (dDAVP), the abundance of pS264-AQP2 increased 4-fold over controls. Additionally, dDAVP treatment resulted in a time dependent change in the distribution of pS264 from predominantly intracellular vesicles, to both the basolateral and apical plasma membranes. Sixty minutes after dDAVP exposure, a proportion of pS264-AQP2 was observed in clathrin-coated vesicles, early endosomal compartments, and recycling compartments, but not lysosomes. Overall, our results are consistent with a dynamic effect of AVP on the phosphorylation and subcellular distribution of AQP2. PMID- 18287044 TI - NEMO recognition of ubiquitinated Bcl10 is required for T cell receptor-mediated NF-kappaB activation. AB - The mechanism by which the Carma1-Bcl10-MALT1 (CBM) complex couples T cell antigen receptor (TCR) signaling to IkappaB kinase (IKK) and NF-kappaB activation is not known. Here, we show that Bcl10 undergoes K63-linked polyubiquitination in response to T cell activation and subsequently binds NEMO, the regulatory subunit of IKK. This interaction requires the ubiquitin-binding activity of NEMO. The sites of Bcl10 ubiquitination were mapped to K31 and K63. Mutation of these residues did not affect TCR signaling-induced CBM complex assembly but prevented Bcl10 ubiquitination, NEMO binding, and NF-kappaB activation. Therefore, the regulated ubiquitination of Bcl10 and its recognition by NEMO are a critical link between the CBM complex, IKK recruitment, and NF-kappaB activation. PMID- 18287045 TI - Dynamics of Pseudomonas aeruginosa genome evolution. AB - One of the hallmarks of the Gram-negative bacterium Pseudomonas aeruginosa is its ability to thrive in diverse environments that includes humans with a variety of debilitating diseases or immune deficiencies. Here we report the complete sequence and comparative analysis of the genomes of two representative P. aeruginosa strains isolated from cystic fibrosis (CF) patients whose genetic disorder predisposes them to infections by this pathogen. The comparison of the genomes of the two CF strains with those of other P. aeruginosa presents a picture of a mosaic genome, consisting of a conserved core component, interrupted in each strain by combinations of specific blocks of genes. These strain-specific segments of the genome are found in limited chromosomal locations, referred to as regions of genomic plasticity. The ability of P. aeruginosa to shape its genomic composition to favor survival in the widest range of environmental reservoirs, with corresponding enhancement of its metabolic capacity is supported by the identification of a genomic island in one of the sequenced CF isolates, encoding enzymes capable of degrading terpenoids produced by trees. This work suggests that niche adaptation is a major evolutionary force influencing the composition of bacterial genomes. Unlike genome reduction seen in host-adapted bacterial pathogens, the genetic capacity of P. aeruginosa is determined by the ability of individual strains to acquire or discard genomic segments, giving rise to strains with customized genomic repertoires. Consequently, this organism can survive in a wide range of environmental reservoirs that can serve as sources of the infecting organisms. PMID- 18287046 TI - Dual regulation by the Hunchback gradient in the Drosophila embryo. AB - The regulation of segmentation gene expression is investigated by computational modeling using quantitative expression data. Previous tissue culture assays and transgene analyses raised the possibility that Hunchback (Hb) might function as both an activator and repressor of transcription. At low concentrations, Hb activates gene expression, whereas at high concentrations it mediates repression. Under the same experimental conditions, transcription factors encoded by other gap genes appear to function as dedicated repressors. Models based on dual regulation suggest that the Hb gradient can be sufficient for establishing the initial Kruppel (Kr) expression pattern in central regions of the precellular embryo. The subsequent refinement of the Kr pattern depends on the combination of Hb and the Giant (Gt) repressor. The dual-regulation models developed for Kr also explain some of the properties of the even-skipped (eve) stripe 3+7 enhancer. Computational simulations suggest that repression results from the dimerization of Hb monomers on the DNA template. PMID- 18287047 TI - PolIVb influences RNA-directed DNA methylation independently of its role in siRNA biogenesis. AB - DNA-dependent RNA polymerase (Pol)IV in Arabidopsis exists in two isoforms (PolIVa and PolIVb), with NRPD1a and NRPD1b as their respective largest subunits. Both isoforms are implicated in production and activity of siRNAs and in RNA directed DNA methylation (RdDM). Deep sequence analysis of siRNAs in WT Arabidopsis flowers and in nrpd1a and nrpd1b mutants identified >4,200 loci producing siRNAs in a PolIV-dependent manner, with PolIVb reinforcing siRNA production by PolIVa. Transposable element identity and pericentromeric localization are both features that predispose a locus for siRNA production via PolIV proteins and determine the extent to which siRNA production relies on PolIVb. Detailed analysis of DNA methylation at PolIV-dependent loci revealed unexpected deviations from the previously noted association of PolIVb-dependent siRNA production and RdDM. Notably, PolIVb functions independently in DNA methylation and siRNA generation. Additionally, we have uncovered siRNA-directed loss of DNA methylation, a process requiring both PolIV isoforms. From these findings, we infer that the role of PolIVb in siRNA production is secondary to a role in chromatin modification and is influenced by chromatin context. PMID- 18287048 TI - Asymmetric segregation of protein aggregates is associated with cellular aging and rejuvenation. AB - Aging, defined as a decrease in reproduction rate with age, is a fundamental characteristic of all living organisms down to bacteria. Yet we know little about the causal molecular mechanisms of aging within the in vivo context of a wild type organism. One of the prominent markers of aging is protein aggregation, associated with cellular degeneracy in many age-related diseases, although its in vivo dynamics and effect are poorly understood. We followed the appearance and inheritance of spontaneous protein aggregation within lineages of Escherichia coli grown under nonstressed conditions using time-lapse microscopy and a fluorescently tagged chaperone (IbpA) involved in aggregate processing. The fluorescent marker is shown to faithfully identify in vivo the localization of aggregated proteins, revealing their accumulation upon cell division in cells with older poles. This accretion is associated with >30% of the loss of reproductive ability (aging) in these cells relative to the new-pole progeny, devoid of parental inclusion bodies, that exhibit rejuvenation. This suggests an asymmetric strategy whereby dividing cells segregate damage at the expense of aging individuals, resulting in the perpetuation of the population. PMID- 18287049 TI - Tpl2 and ERK transduce antiproliferative T cell receptor signals and inhibit transformation of chronically stimulated T cells. AB - The protein kinase encoded by the Tpl2 protooncogene plays an obligatory role in the transduction of Toll-like receptor and death receptor signals in macrophages, B cells, mouse embryo fibroblasts, and epithelial cells in culture and promotes inflammatory responses in animals. To address its role in T cell activation, we crossed the T cell receptor (TCR) transgene 2C, which recognizes class I MHC presented peptides, into the Tpl2(-/-) genetic background. Surprisingly, the TCR2C(tg/tg)/Tpl2(-/-) mice developed T cell lymphomas with a latency of 4-6 months. The tumor cells were consistently TCR2C(+)CD8(+)CD4(-), suggesting that they were derived either from chronically stimulated mature T cells or from immature single positive (ISP) cells. Further studies showed that the population of CD8(+) ISP cells was not expanded in the thymus of TCR2C(tg/tg)/Tpl2(-/-) mice, making the latter hypothesis unlikely. Mature peripheral T cells of Tpl2(-/ ) mice were defective in ERK activation and exhibited enhanced proliferation after TCR stimulation. The same cells were defective in the induction of CTLA4, a negative regulator of the T cell response, which is induced by TCR signals via ERK. These findings suggest that Tpl2 functions normally in a feedback loop that switches off the T cell response to TCR stimulation. As a result, Tpl2, a potent oncogene, functions as a tumor suppressor gene in chronically stimulated T cells. PMID- 18287050 TI - DNA barcodes: genes, genomics, and bioinformatics. PMID- 18287051 TI - Ominous trends in nature recreation. PMID- 18287052 TI - miR-17-92 cluster accelerates adipocyte differentiation by negatively regulating tumor-suppressor Rb2/p130. AB - Adipogenesis involves cell proliferation and differentiation, both of which have been shown to be regulated by micro (mi)RNA. During mouse preadipocyte 3T3L1 cell differentiation, we found that miR-17-92, a miRNA cluster that promotes cell proliferation in various cancers, was significantly up-regulated at the clonal expansion stage of adipocyte differentiation. Stable transfection of 3T3L1 cells with miR-17-92 resulted in accelerated differentiation and increased triglyceride accumulation after hormonal stimulation. By using a luciferase reporter assay, we demonstrated that miR-17-92 directly targeted the 3' UTR region of Rb2/p130, accounting for subsequently reduced Rb2/p130 mRNA and protein quantities at the stage of clonal expansion. siRNA-mediated knock-down of Rb2/p130 at the same stage of clonal expansion recapitulated the phenotype of overexpression of miR-17 92 in the stably transfected 3T3L1 cells. These data indicate that miR-17-92 promotes adipocyte differentiation by targeting and negatively regulating Rb2/p130. PMID- 18287053 TI - Inhibition of proinflammatory and innate immune signaling pathways by a cytomegalovirus RIP1-interacting protein. AB - TNFalpha is an important cytokine in antimicrobial immunity and inflammation. The receptor-interacting protein RIP1 is an essential component of the TNF receptor 1 signaling pathway that mediates the activation of NF-kappaB, MAPKs, and programmed cell death. It also transduces signals derived from Toll-like receptors and intracellular sensors of DNA damage and double-stranded RNA. Here, we show that the murine CMV M45 protein binds to RIP1 and inhibits TNFalpha induced activation of NF-kappaB, p38 MAPK, and caspase-independent cell death. M45 also inhibited NF-kappaB activation upon stimulation of Toll-like receptor 3 and ubiquitination of RIP1, which is required for NF-kappaB activation. Hence, M45 functions as a viral inhibitor of RIP1-mediated signaling. The results presented here reveal a mechanism of viral immune subversion and demonstrate how a viral protein can simultaneously block proinflammatory and innate immune signaling pathways by interacting with a central mediator molecule. PMID- 18287054 TI - A folding space odyssey. PMID- 18287055 TI - Synapse elimination accompanies functional plasticity in hippocampal neurons. AB - A critical component of nervous system development is synapse elimination during early postnatal life, a process known to depend on neuronal activity. Changes in synaptic strength in the form of long-term potentiation (LTP) and long-term depression (LTD) correlate with dendritic spine enlargement or shrinkage, respectively, but whether LTD can lead to an actual separation of the synaptic structures when the spine shrinks or is lost remains unknown. Here, we addressed this issue by using concurrent imaging and electrophysiological recording of live synapses. Slices of rat hippocampus were cultured on multielectrode arrays, and the neurons were labeled with genes encoding red or green fluorescent proteins to visualize presynaptic and postsynaptic neuronal processes, respectively. LTD inducing stimulation led to a reduction in the synaptic green and red colocalization, and, in many cases, it induced a complete separation of the presynaptic bouton from the dendritic spine. This type of synapse loss was associated with smaller initial spine size and greater synaptic depression but not spine shrinkage during LTD. All cases of synapse separation were observed without an accompanying loss of the spine during this period. We suggest that repeated low-frequency stimulation simultaneous with LTD induction is capable of restructuring synaptic contacts. Future work with this model will be able to provide critical insight into the molecular mechanisms of activity- and experience-dependent refinement of brain circuitry during development. PMID- 18287056 TI - Protein area occupancy at the center of the red blood cell membrane. AB - In the Fluid Mosaic Model for biological membrane structure, proposed by Singer and Nicolson in 1972, the lipid bilayer is represented as a neutral two dimensional solvent in which the proteins of the membrane are dispersed and distributed randomly. The model portrays the membrane as dominated by a membrane lipid bilayer, directly exposed to the aqueous environment, and only occasionally interrupted by transmembrane proteins. This view is reproduced in virtually every textbook in biochemistry and cell biology, yet some critical features have yet to be closely examined, including the key parameter of the relative occupancy of protein and lipid at the center of a natural membrane. Here we show that the area occupied by protein and lipid at the center of the human red blood cell (RBC) plasma membrane is at least approximately 23% protein and less than approximately 77% lipid. This measurement is in close agreement with previous estimates for the RBC plasma membrane and the recently published measurements for the synaptic vesicle. Given that transmembrane proteins are surrounded by phospholipids that are perturbed by their presence, the occupancy by protein of more than approximately 20% of the RBC plasma membrane and the synaptic vesicle plasma membrane implies that natural membrane bilayers may be more rigid and less fluid than has been thought for the past several decades, and that studies of pure lipid bilayers do not fully reveal the properties of lipids in membranes. Thus, it appears to be the case that membranes may be more mosaic than fluid, with little unperturbed phospholipid bilayer. PMID- 18287057 TI - Retinoic acid deficiency alters second heart field formation. AB - Retinoic acid (RA), the active derivative of vitamin A, has been implicated in various steps of cardiovascular development. The retinaldehyde dehydrogenase 2 (RALDH2) enzyme catalyzes the second oxidative step in RA biosynthesis and its loss of function creates a severe embryonic RA deficiency. Raldh2(-/-) knockout embryos fail to undergo heart looping and have impaired atrial and sinus venosus development. To understand the mechanism(s) producing these changes, we examined the contribution of the second heart field (SHF) to pharyngeal mesoderm, atria, and outflow tract in Raldh2(-/-) embryos. RA deficiency alters SHF gene expression in two ways. First, Raldh2(-/-) embryos exhibited a posterior expansion of anterior markers of the SHF, including Tbx1, Fgf8, and the Mlc1v nlacZ-24/Fgf10 reporter transgene as well as of Islet1. This occurred at early somite stages, when cardiac defects became irreversible in an avian vitamin A deficiency model, indicating that endogenous RA is required to restrict the SHF posteriorly. Explant studies showed that this expanded progenitor population cannot differentiate properly. Second, RA up-regulated cardiac Bmp expression levels at the looping stage. The contribution of the SHF to both inflow and outflow poles was perturbed under RA deficiency, creating a disorganization of the heart tube. We also investigated genetic cross-talk between Nkx2.5 and RA signaling by generating double mutant mice. Strikingly, Nkx2.5 deficiency was able to rescue molecular defects in the posterior region of the Raldh2(-/-) mutant heart, in a gene dosage-dependent manner. PMID- 18287058 TI - A systems biology approach identifies the biochemical mechanisms regulating monoterpenoid essential oil composition in peppermint. AB - The integration of mathematical modeling and experimental testing is emerging as a powerful approach for improving our understanding of the regulation of metabolic pathways. In this study, we report on the development of a kinetic mathematical model that accurately simulates the developmental patterns of monoterpenoid essential oil accumulation in peppermint (Mentha x piperita). This model was then used to evaluate the biochemical processes underlying experimentally determined changes in the monoterpene pathway under low ambient light intensities, which led to an accumulation of the branchpoint intermediate (+)-pulegone and the side product (+)-menthofuran. Our simulations indicated that the environmentally regulated changes in monoterpene profiles could only be explained when, in addition to effects on biosynthetic enzyme activities, as yet unidentified inhibitory effects of (+)-menthofuran on the branchpoint enzyme pulegone reductase (PR) were assumed. Subsequent in vitro analyses with recombinant protein confirmed that (+)-menthofuran acts as a weak competitive inhibitor of PR (K(i) = 300 muM). To evaluate whether the intracellular concentration of (+)-menthofuran was high enough for PR inhibition in vivo, we isolated essential oil-synthesizing secretory cells from peppermint leaves and subjected them to steam distillations. When peppermint plants were grown under low-light conditions, (+)-menthofuran was selectively retained in secretory cells and accumulated to very high levels (up to 20 mM), whereas under regular growth conditions, (+)-menthofuran levels remained very low (<400 muM). These results illustrate the utility of iterative cycles of mathematical modeling and experimental testing to elucidate the mechanisms controlling flux through metabolic pathways. PMID- 18287059 TI - Elevated globotriaosylsphingosine is a hallmark of Fabry disease. AB - Fabry disease is an X-linked lysosomal storage disease caused by deficiency of alpha-galactosidase A that affects males and shows disease expression in heterozygotes. The characteristic progressive renal insufficiency, cardiac involvement, and neuropathology usually are ascribed to globotriaosylceramide accumulation in the endothelium. However, no direct correlation exists between lipid storage and clinical manifestations, and treatment of patients with recombinant enzymes does not reverse several key signs despite clearance of lipid from the endothelium. We therefore investigated the possibility that globotriaosylceramide metabolites are a missing link in the pathogenesis. We report that deacylated globotriaosylceramide, globotriaosylsphingosine, and a minor additional metabolite are dramatically increased in plasma of classically affected male Fabry patients and plasma and tissues of Fabry mice. Plasma globotriaosylceramide levels are reduced by therapy. We show that globotriaosylsphingosine is an inhibitor of alpha-galactosidase A activity. Furthermore, exposure of smooth muscle cells, but not fibroblasts, to globotriaosylsphingosine at concentrations observed in plasma of patients promotes proliferation. The increased intima-media thickness in Fabry patients therefore may be related to the presence of this metabolite. Our findings suggest that measurement of circulating globotriaosylsphingosine will be useful to monitor Fabry disease and may contribute to a better understanding of the disorder. PMID- 18287060 TI - Ultrasonic vocalization impairment of Foxp2 (R552H) knockin mice related to speech-language disorder and abnormality of Purkinje cells. AB - Previous studies have demonstrated that mutation in the forkhead domain of the forkhead box P2 (FOXP2) protein (R553H) causes speech-language disorders. To further analyze FOXP2 function in speech learning, we generated a knockin (KI) mouse for Foxp2 (R552H) [Foxp2 (R552H)-KI], corresponding to the human FOXP2 (R553H) mutation, by homologous recombination. Homozygous Foxp2 (R552H)-KI mice showed reduced weight, immature development of the cerebellum with incompletely folded folia, Purkinje cells with poor dendritic arbors and less synaptophysin immunoreactivity, and achieved crisis stage for survival 3 weeks after birth. At postnatal day 10, these mice also showed severe ultrasonic vocalization (USV) and motor impairment, whereas the heterozygous Foxp2 (R552H)-KI mice exhibited modest impairments. Similar to the wild-type protein, Foxp2 (R552H) localized in the nuclei of the Purkinje cells and the thalamus, striatum, cortex, and hippocampus (CA1) neurons of the homozygous Foxp2 (R552H)-KI mice (postnatal day 10), and some of the neurons showed nuclear aggregates of Foxp2 (R552H). In addition to the immature development of the cerebellum, Foxp2 (R552H) nuclear aggregates may further compromise the function of the Purkinje cells and cerebral neurons of the homozygous mice, resulting in their death. In contrast, heterozygous Foxp2 (R552H)-KI mice, which showed modest impairment of USVs with different USV qualities and which did not exhibit nuclear aggregates, should provide insights into the common molecular mechanisms between the mouse USV and human speech learning and the relationship between the USV and motor neural systems. PMID- 18287061 TI - Functional interactions among Orai1, TRPCs, and STIM1 suggest a STIM-regulated heteromeric Orai/TRPC model for SOCE/Icrac channels. AB - Receptor-operated Ca(2+) entry (ROCE) and store-operated Ca(2+) entry (SOCE) into cells are functions performed by all higher eukaryotic cells, and their impairment is life-threatening. The main molecular components of this pathway appear to be known. However, the molecular make-up of channels mediating ROCE and SOCE is largely unknown. One hypothesis proposes SOCE channels to be formed solely by Orai proteins. Another proposes SOCE channels to be composed of both Orai and C-type transient receptor potential (TRPC) proteins. Both hypotheses propose that the channels are activated by STIM1, a sensor of the filling state of the Ca(2+) stores that activates Ca(2+) entry when stores are depleted. The role of Orai in SOCE has been proven. Here we show the TRPC-dependent reconstitution of Icrac, the electrophysiological correlate to SOCE, by expression of Orai1; we also show that R91W-Orai1 can inhibit SOCE and ROCE and that Orai1 and STIM1 expression leads to functional expression of Gd-resistant ROCE. Because channels that mediate ROCE are accepted to be formed with the participation of TRPCs, our data show functional interaction between ROCE and SOCE components. We propose that SOCE/Icrac channels are composed of heteromeric complexes that include TRPCs and Orai proteins. PMID- 18287062 TI - Immunologic and clinical effects of antibody blockade of cytotoxic T lymphocyte associated antigen 4 in previously vaccinated cancer patients. AB - Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) functions as a negative regulator of endogenous and vaccine-induced antitumor immunity. The administration of fully human anti-CTLA-4 blocking monoclonal antibodies to advanced-cancer patients increases immune-mediated tumor destruction in some subjects. Nonetheless, patients that respond also frequently manifest serious inflammatory pathologies, raising the possibility that the therapeutic and toxic effects of CTLA-4 blockade might be linked. Here we show that periodic infusions of anti-CTLA-4 antibodies after vaccination with irradiated, autologous tumor cells engineered to secrete GM-CSF (GVAX) generate clinically meaningful antitumor immunity without grade 3 or 4 toxicity in a majority of metastatic melanoma patients. The application of this sequential immunotherapy to advanced ovarian carcinoma patients also revealed that tumor destruction and severe inflammatory pathology could be dissociated, although further refinements are required to increase clinical responses and to minimize toxicity in this population. The extent of therapy-induced tumor necrosis was linearly related to the natural logarithm of the ratio of intratumoral CD8(+) effector T cells to FoxP3(+) regulatory T cells (Tregs) in posttreatment biopsies. Together, these findings help clarify the immunologic and clinical effects of CTLA-4 antibody blockade in previously vaccinated patients and raise the possibility that selective targeting of antitumor Tregs may constitute a complementary strategy for combination therapy. PMID- 18287063 TI - HLA alleles determine differences in human natural killer cell responsiveness and potency. AB - Epidemiological studies have associated certain human disease outcomes with particular killer cell Ig-like receptor (KIR) and HLA genotypes. However, the functional explanation for these associations is poorly understood, because the KIRs were initially described as natural killer (NK) cell inhibitory receptors with specificity for HLA molecules on their cellular targets. Yet resolution of infections is often associated with genotypic pairing of inhibitory KIRs with their cognate HLA ligands. Recent studies in mice indicate a second role for MHC specific inhibitory receptors, i.e., self-MHC recognition confers functional competence on the NK cell to be triggered through their activation receptors, a process termed licensing. As a result, licensed NK cells with self-MHC-specific receptors are more readily activated as compared with unlicensed NK cells without self-MHC-specific receptors. Such results predict that human NK cells may undergo a similar process. Here, we examined the human NK cell subset expressing KIR3DL1, the only known KIR specific for HLA-Bw4 alleles. The KIR3DL1(+) subset in normal donors with two HLA-B-Bw4 genes displayed increased responsiveness to tumor stimulation compared with the KIR3DL1(+) subset from individuals with only one or no Bw4 genes. By contrast, NK cells lacking KIR3DL1 showed no differences. Therefore, these data indicate that particular KIR and HLA alleles are associated with more responsive NK cells, strongly suggesting that human NK cells are also subjected to NK cell licensing, and providing a potential functional explanation for the influence of KIR and HLA genes in disease as well as interindividual differences in NK cell potency. PMID- 18287064 TI - HSP90-buffered genetic variation is common in Arabidopsis thaliana. AB - HSP90 is a protein chaperone particularly important in the maturation of a diverse set of proteins that regulate key steps in a multitude of biological processes. Alterations in HSP90 function produce altered phenotypes at low penetrance in natural populations. Previous work has shown that at least some of these phenotypes are due to genetic variation that remains phenotypically cryptic until it is revealed by the impairment of HSP90 function. Exposure of such "buffered" genetic polymorphisms can also be accomplished by environmental stress, linking the appearance of new phenotypes to defects in protein homeostasis. Should such polymorphisms be widespread, natural selection may be more effective at producing phenotypic change in suboptimal environments. In evaluating this hypothesis, a key unknown factor is the frequency with which HSP90-buffered polymorphisms occur in natural populations. Here, we present Arabidopsis thaliana populations suitable for genetic mapping that have constitutively reduced HSP90 levels. We employ quantitative genetic techniques to examine the HSP90-dependent polymorphisms affecting a host of plastic plant life history traits. Our results demonstrate that HSP90-dependent natural variation is present at high frequencies in A. thaliana, with an expectation that at least one HSP90-dependent polymorphism will affect nearly every quantitative trait in progeny of two different wild lines. Hence, HSP90 is likely to occupy a central position in the translation of genotypic variation into phenotypic differences. PMID- 18287065 TI - HSP90 affects the expression of genetic variation and developmental stability in quantitative traits. AB - Modulation of the activity of the molecular chaperone HSP90 has been extensively discussed as a means to alter phenotype in many traits and organisms. Such changes can be due to the exposure of cryptic genetic variation, which in some instances may also be accomplished by mild environmental alteration. Should such polymorphisms be widespread, natural selection may be more effective at producing phenotypic change in suboptimal environments. However, the frequency and identity of buffered polymorphisms in natural populations are unknown. Here, we employ quantitative genetic dissection of an Arabidopsis thaliana developmental response, hypocotyl elongation in the dark, to detail the underpinnings of genetic variation responsive to HSP90 modulation. We demonstrate that HSP90 dependent alleles occur in continuously distributed, environmentally responsive traits and are amenable to quantitative genetic mapping techniques. Furthermore, such alleles are frequent in natural populations and can have significant effects on natural phenotypic variation. We also find that HSP90 modulation has both general and allele-specific effects on developmental stability; that is, developmental stability is a phenotypic trait that can be affected by natural variation. However, effects of revealed variation on trait means outweigh effects of decreased developmental stability, and the HSP90-dependent trait alterations could be acted on by natural selection. Thus, HSP90 may centrally influence canalization, assimilation, and the rapid evolutionary alteration of phenotype through the concealment and exposure of cryptic genetic variation. PMID- 18287066 TI - The Foldback-like element Galileo belongs to the P superfamily of DNA transposons and is widespread within the Drosophila genus. AB - Galileo is the only transposable element (TE) known to have generated natural chromosomal inversions in the genus Drosophila. It was discovered in Drosophila buzzatii and classified as a Foldback-like element because of its long, internally repetitive, terminal inverted repeats (TIRs) and lack of coding capacity. Here, we characterized a seemingly complete copy of Galileo from the D. buzzatii genome. It is 5,406 bp long, possesses 1,229-bp TIRs, and encodes a 912 aa transposase similar to those of the Drosophila melanogaster 1360 (Hoppel) and P elements. We also searched the recently available genome sequences of 12 Drosophila species for elements similar to Dbuz?Galileo by using bioinformatic tools. Galileo was found in six species (ananassae, willistoni, peudoobscura, persimilis, virilis, and mojavensis) from the two main lineages within the Drosophila genus. Our observations place Galileo within the P superfamily of cut and-paste transposons and extend considerably its phylogenetic distribution. The interspecific distribution of Galileo indicates an ancient presence in the genus, but the phylogenetic tree built with the transposase amino acid sequences contrasts significantly with that of the species, indicating lineage sorting and/or horizontal transfer events. Our results also suggest that Foldback-like elements such as Galileo may evolve from DNA-based transposon ancestors by loss of the transposase gene and disproportionate elongation of TIRs. PMID- 18287068 TI - Quantitative biochemical rationale for differences in transmissibility of 1918 pandemic influenza A viruses. AB - The human adaptation of influenza A viruses is critically governed by the binding specificity of the viral surface hemagglutinin (HA) to long (chain length) alpha2 6 sialylated glycan (alpha2-6) receptors on the human upper respiratory tissues. A recent study demonstrated that whereas the 1918 H1N1 pandemic virus, A/South Carolina/1/1918 (SC18), with alpha2-6 binding preference transmitted efficiently, a single amino acid mutation on HA resulted in a mixed alpha2-3 sialylated glycan (alpha2-3)/alpha2-6 binding virus (NY18) that transmitted inefficiently. To define the biochemical basis for the observed differences in virus transmission, in this study, we have developed an approach to quantify the multivalent HA glycan interactions. Analysis of the molecular HA-glycan contacts showed subtle changes resulting from the single amino acid variations between SC18 and NY18. The effect of these changes on glycan binding is amplified by multivalency, resulting in quantitative differences in their long alpha2-6 glycan binding affinities. Furthermore, these differences are also reflected in the markedly distinct binding pattern of SC18 and NY18 HA to the physiological glycans present in human upper respiratory tissues. Thus, the dramatic lower binding affinity of NY18 to long alpha2-6 glycans, as against a mixed alpha2-3/6 binding, correlates with its inefficient transmission. In summary, this study establishes a quantitative biochemical correlate for influenza A virus transmission. PMID- 18287069 TI - Single gene reassortants identify a critical role for PB1, HA, and NA in the high virulence of the 1918 pandemic influenza virus. AB - The 1918 influenza pandemic was exceptionally severe, resulting in the death of up to 50 million people worldwide. Here, we show which virus genes contributed to the replication and virulence of the 1918 influenza virus. Recombinant viruses, in which genes of the 1918 virus were replaced with genes from a contemporary human H1N1 influenza virus, A/Texas/36/91 (Tx/91), were generated. The exchange of most 1918 influenza virus genes with seasonal influenza H1N1 virus genes did not alter the virulence of the 1918 virus; however, substitution of the hemagglutinin (HA), neuraminidase (NA), or polymerase subunit PB1 genes significantly affected the ability of this virus to cause severe disease in mice. The 1918 virus virulence observed in mice correlated with the ability of 1918 recombinant viruses to replicate efficiently in human airway cells. In a second series of experiments, eight 1918 1:7 recombinants were generated, in which each Tx/91 virus gene was individually replaced by a corresponding gene from 1918 virus. Replication capacity of the individual 1:7 reassortant viruses was assessed in mouse lungs and human airway cells. Increased virus titers were observed among 1:7 viruses containing individual 1918 HA, NA, and PB1 genes. In addition, the 1918 PB1:Tx/91 (1:7) virus showed a distinctly larger plaque size phenotype than the small plaque phenotype of the 1918 PA:Tx/91 and 1918 PB2:Tx/91 1:7 reassortants. These results highlight the importance of the 1918 HA, NA, and PB1 genes for optimal virus replication and virulence of this pandemic strain. PMID- 18287070 TI - Quorum size of Pseudomonas syringae is small and dictated by water availability on the leaf surface. AB - The paradigm of bacterial quorum sensing (QS), which mediates cell-density dependent gene expression, usually has been studied in high-cell-density planktonic liquid cultures or in biofilms in which signal concentrations accumulate to sufficiently high levels to induce QS. Presumably under conditions with restricted diffusion of the signal molecule, smaller population sizes could achieve such a state of QS induction. The plant-pathogenic bacterium Pseudomonas syringae, in which QS controls traits involved in epiphytic fitness and virulence, occurs on leaf surfaces in aggregates of various sizes. Because leaves often harbor limited surface water, we investigated the size of aggregates that would permit QS in a nonsaturated environment. QS induction was visualized via dual fluorescence of P. syringae cells harboring a transcriptional fusion of mRFP1 with ahlI, which exhibits N-acyl homoserine lactone-dependent transcriptional activity, and a constitutive GFP marker to account for all P. syringae cells on a leaf. Confocal microscopy revealed that, on wet leaves, no QS induction was evident within 2 days after inoculation, but it increased rapidly with increasing aggregate sizes >40 and 22 cells per aggregate by 3 and 4 days, respectively. In contrast, QS induction was common in aggregates >33 cells by 2 days after inoculation on dry leaves and increased rapidly with increasing aggregate sizes >35 and 13 cells after 3 and 4 days, respectively. These observations demonstrate that small groups of cells experience QS conditions on dry leaves where signal diffusion is restricted. Quorum size of bacteria in non water-saturated environments such as on leaves is small, and QS induction may be commonly operative. PMID- 18287072 TI - PDC-TREM, a plasmacytoid dendritic cell-specific receptor, is responsible for augmented production of type I interferon. AB - Type I interferons (IFNs) derived from plasmacytoid dendritic cells (PDCs) are critical for antiviral responses; however, the mechanisms underlying their production remain unclear. We have identified a receptor, PDC-TREM, which is associated with Plexin-A1 (PlxnA1) on the PDC cell surface and is preferentially expressed after TLR-stimulation. Limited TLR signals induced PDC-TREM expression but failed to induce IFN-alpha production. However, when coupled with Sema6D, a ligand for Plexin-A1, limited TLR-stimulation resulted in PDC-TREM-mediated DAP12 dependent phosphorylation of phosphoinositide 3-kinase (PI3K) and extracellular regulated kinase (Erk) 1/2 at 6-9 h, and IFN-alpha was produced. Inhibition of PDC-TREM expression by pdctrem-shRNA, blocking of PDC-TREM-binding with PlxnA1 by PDC-TREM mAb, and DAP12 deficiency all resulted in greatly reduced PDC-TREM dependent activation of signaling molecules and IFN-alpha production. Thus, PDC TREM is responsible for IFN-alpha production, whereas TLR signals are essential for PDC-TREM expression. PMID- 18287073 TI - A systems approach to delineate functions of paralogous transcription factors: role of the Yap family in the DNA damage response. AB - Duplication of genes encoding transcription factors plays an essential role in driving phenotypic variation. Because regulation can occur at multiple levels, it is often difficult to discern how each duplicated factor achieves its regulatory specificity. In these cases, a "systems approach" may distinguish the role of each factor by integrating complementary large-scale measurements of the regulatory network. To explore such an approach, we integrate growth phenotypes, promoter binding profiles, and gene expression patterns to model the DNA damage response network controlled by the Yeast-specific AP-1 (YAP) family of transcription factors. This analysis reveals that YAP regulatory specificity is achieved by at least three mechanisms: (i) divergence of DNA-binding sequences into two subfamilies; (ii) condition-specific combinatorial regulation by multiple Yap factors; and (iii) interactions of Yap 1, 4, and 6 with chromatin remodeling proteins. Additional microarray experiments establish that Yap 4 and 6 regulate gene expression through interactions with the histone deacetylase, Hda1. The data further highlight differences among Yap paralogs in terms of their regulatory mode of action (activation vs. repression). This study suggests how other large TF families might be disentangled in the future. PMID- 18287074 TI - Regulation of excitability and plasticity by endocannabinoids and PKA in developing hippocampus. AB - The activity-dependent strengthening and weakening of synaptic transmission are hypothesized to be the basis of not only memory and learning but also the refinement of neural circuits during development. Here we report that, in the developing CA1 area of the hippocampus, endocannabinoid (eCB)-mediated heterosynaptic long-term depression (LTD) of glutamatergic excitatory synaptic transmission is associated with PKA-mediated homosynaptic long-term potentiation (LTP). This form of LTD was dominant at postnatal days 2-10 (P2-P10), attenuated during development, and finally disappeared in the mature hippocampus. Heterosynaptic LTD of excitatory postsynaptic currents in the developing hippocampus was expressed presynaptically, spread to neighboring neurons, and was mediated by eCBs. Heterosynaptic LTD of field excitatory postsynaptic potentials was associated with a decrease in fiber volley amplitude with a similar time course. Depression of fiber volleys was blocked by K(+) channel blockers, suggesting the involvement of the decrease in presynaptic excitability in heterosynaptic LTD. In the P2-P5 hippocampus, eCBs also attenuate LTP and fiber volleys in homosynaptic pathways and help to prevent too much excitability in the neonatal hippocampus where the GABAergic system is poorly developed and even excitatory. In the hippocampus older than P6 (P > 6), however, LTP is protected from eCB-mediated depression by PKA activated at presynaptic sites by high frequency stimulation, serving to highlight PKA-mediated LTP by weakening inactive synapses even in adjacent cells. Thus, eCBs and PKA make synapses plastic without changing excitability homeostasis in the developing hippocampus. PMID- 18287075 TI - Structure of a protein-DNA complex essential for DNA protection in spores of Bacillus species. AB - The DNA-binding alpha/beta-type small acid-soluble proteins (SASPs) are a major factor in the resistance and long-term survival of spores of Bacillus species by protecting spore DNA against damage due to desiccation, heat, toxic chemicals, enzymes, and UV radiation. We now report the crystal structure at 2.1 A resolution of an alpha/beta-type SASP bound to a 10-bp DNA duplex. In the complex, the alpha/beta-type SASP adopt a helix-turn-helix motif, interact with DNA through minor groove contacts, bind to approximately 6 bp of DNA as a dimer, and the DNA is in an A-B type conformation. The structure of the complex provides important insights into the molecular details of both DNA and alpha/beta-type SASP protection in the complex and thus also in spores. PMID- 18287076 TI - A giant frog with South American affinities from the Late Cretaceous of Madagascar. AB - Madagascar has a diverse but mainly endemic frog fauna, the biogeographic history of which has generated intense debate, fueled by recent molecular phylogenetic analyses and the near absence of a fossil record. Here, we describe a recently discovered Late Cretaceous anuran that differs strikingly in size and morphology from extant Malagasy taxa and is unrelated either to them or to the predicted occupants of the Madagascar-Seychelles-India landmass when it separated from Africa 160 million years ago (Mya). Instead, the previously undescribed anuran is attributed to the Ceratophryinae, a clade previously considered endemic to South America. The discovery offers a rare glimpse of the anuran assemblage that occupied Madagascar before the Tertiary radiation of mantellids and microhylids that now dominate the anuran fauna. In addition, the presence of a ceratophryine provides support for a controversial paleobiogeographical model that posits physical and biotic links among Madagascar, the Indian subcontinent, and South America that persisted well into the Late Cretaceous. It also suggests that the initial radiation of hyloid anurans began earlier than proposed by some recent estimates. PMID- 18287077 TI - Generation of human induced pluripotent stem cells from dermal fibroblasts. AB - The generation of patient-specific pluripotent stem cells has the potential to accelerate the implementation of stem cells for clinical treatment of degenerative diseases. Technologies including somatic cell nuclear transfer and cell fusion might generate such cells but are hindered by issues that might prevent them from being used clinically. Here, we describe methods to use dermal fibroblasts easily obtained from an individual human to generate human induced pluripotent stem (iPS) cells by ectopic expression of the defined transcription factors KLF4, OCT4, SOX2, and C-MYC. The resultant cell lines are morphologically indistinguishable from human embryonic stem cells (HESC) generated from the inner cell mass of a human preimplantation embryo. Consistent with these observations, human iPS cells share a nearly identical gene-expression profile with two established HESC lines. Importantly, DNA fingerprinting indicates that the human iPS cells were derived from the donor material and are not a result of contamination. Karyotypic analyses demonstrate that reprogramming of human cells by defined factors does not induce, or require, chromosomal abnormalities. Finally, we provide evidence that human iPS cells can be induced to differentiate along lineages representative of the three embryonic germ layers indicating the pluripotency of these cells. Our findings are an important step toward manipulating somatic human cells to generate an unlimited supply of patient specific pluripotent stem cells. In the future, the use of defined factors to change cell fate may be the key to routine nuclear reprogramming of human somatic cells. PMID- 18287078 TI - SOX2-expressing progenitor cells generate all of the major cell types in the adult mouse pituitary gland. AB - The pituitary gland adapts the proportion of each of its endocrine cell types to meet differing hormonal demands throughout life. There is circumstantial evidence that multipotent adult progenitor cells contribute to this plasticity, but these cells have not been identified. Here, we describe a small (<0.05%) population of progenitor cells in the adult pituitary gland. We show that these cells express SOX2, a marker of several early embryonic progenitor and stem cell types, and form "pituispheres" in culture, which can grow, form secondary spheres, and differentiate to all of the pituitary endocrine cell types, as well as folliculostellate cells. Differentiation of cells in the pituispheres was associated with the expression of nestin, SOX9, and S100. Cells expressing SOX2 and E-cadherin are found throughout Rathke's pouch (RP) in embryos but persist in the adult gland, mostly in a narrow zone lining the pituitary cleft, but also are scattered throughout the pituitary. However, unlike in embryonic RP, most of these SOX2(+) cells in the adult gland also express SOX9 and S100. We suggest that this SOX2(+)/SOX9(+) population represents transit-amplifying cells, whereas the SOX2(+)/SOX9(-) cells we identify are multipotent progenitor/stem cells persisting in the adult pituitary. PMID- 18287079 TI - Gambling for global goods. PMID- 18287080 TI - Zero-field remote detection of NMR with a microfabricated atomic magnetometer. AB - We demonstrate remote detection of nuclear magnetic resonance (NMR) with a microchip sensor consisting of a microfluidic channel and a microfabricated vapor cell (the heart of an atomic magnetometer). Detection occurs at zero magnetic field, which allows operation of the magnetometer in the spin-exchange relaxation free (SERF) regime and increases the proximity of sensor and sample by eliminating the need for a solenoid to create a leading field. We achieve pulsed NMR linewidths of 26 Hz, limited, we believe, by the residence time and flow dispersion in the encoding region. In a fully optimized system, we estimate that for 1 s of integration, 7 x 10(13) protons in a volume of 1 mm(3), prepolarized in a 10-kG field, can be detected with a signal-to-noise ratio of approximately 3. This level of sensitivity is competitive with that demonstrated by microcoils in 100-kG magnetic fields, without requiring superconducting magnets. PMID- 18287081 TI - The collective-risk social dilemma and the prevention of simulated dangerous climate change. AB - Will a group of people reach a collective target through individual contributions when everyone suffers individually if the target is missed? This "collective-risk social dilemma" exists in various social scenarios, the globally most challenging one being the prevention of dangerous climate change. Reaching the collective target requires individual sacrifice, with benefits to all but no guarantee that others will also contribute. It even seems tempting to contribute less and save money to induce others to contribute more, hence the dilemma and the risk of failure. Here, we introduce the collective-risk social dilemma and simulate it in a controlled experiment: Will a group of people reach a fixed target sum through successive monetary contributions, when they know they will lose all their remaining money with a certain probability if they fail to reach the target sum? We find that, under high risk of simulated dangerous climate change, half of the groups succeed in reaching the target sum, whereas the others only marginally fail. When the risk of loss is only as high as the necessary average investment or even lower, the groups generally fail to reach the target sum. We conclude that one possible strategy to relieve the collective-risk dilemma in high-risk situations is to convince people that failure to invest enough is very likely to cause grave financial loss to the individual. Our analysis describes the social window humankind has to prevent dangerous climate change. PMID- 18287082 TI - A biodegradable and biocompatible gecko-inspired tissue adhesive. AB - There is a significant medical need for tough biodegradable polymer adhesives that can adapt to or recover from various mechanical deformations while remaining strongly attached to the underlying tissue. We approached this problem by using a polymer poly(glycerol-co-sebacate acrylate) and modifying the surface to mimic the nanotopography of gecko feet, which allows attachment to vertical surfaces. Translation of existing gecko-inspired adhesives for medical applications is complex, as multiple parameters must be optimized, including: biocompatibility, biodegradation, strong adhesive tissue bonding, as well as compliance and conformability to tissue surfaces. Ideally these adhesives would also have the ability to deliver drugs or growth factors to promote healing. As a first demonstration, we have created a gecko-inspired tissue adhesive from a biocompatible and biodegradable elastomer combined with a thin tissue-reactive biocompatible surface coating. Tissue adhesion was optimized by varying dimensions of the nanoscale pillars, including the ratio of tip diameter to pitch and the ratio of tip diameter to base diameter. Coating these nanomolded pillars of biodegradable elastomers with a thin layer of oxidized dextran significantly increased the interfacial adhesion strength on porcine intestine tissue in vitro and in the rat abdominal subfascial in vivo environment. This gecko-inspired medical adhesive may have potential applications for sealing wounds and for replacement or augmentation of sutures or staples. PMID- 18287083 TI - Nrf2 mediates cancer protection but not prolongevity induced by caloric restriction. AB - Caloric restriction (CR) is the most potent intervention known to both protect against carcinogenesis and extend lifespan in laboratory animals. A variety of anticarcinogens and CR mimetics induce and activate the NF-E2-related factor 2 (Nrf2) pathway. Nrf2, in turn, induces a number of antioxidative and carcinogen detoxifying enzymes. Thus, Nrf2 offers a promising target for anticarcinogenesis and antiaging interventions. We used Nrf2-disrupted (KO) mice to examine its role on the biological effects of CR. Here, we show that Nrf2 is responsible for most of the anticarcinogenic effects of CR, but is dispensable for increased insulin sensitivity and lifespan extension. Nrf2-deficient mice developed tumors more readily in response to carcinogen exposure than did WT mice, and CR was ineffective in suppressing tumors in the KO mice. However, CR extended lifespan and increased insulin sensitivity similarly in KO and WT mice. These findings identify a molecular pathway that dissociates the prolongevity and anticarcinogenic effects of CR. PMID- 18287084 TI - Arterial calcifications and increased expression of vitamin D receptor targets in mice lacking TIF1alpha. AB - Calcification of arteries is a major risk factor for cardiovascular mortality in humans. Using genetic approaches, we demonstrate here that the transcriptional intermediary factor 1alpha (TIF1alpha), recently shown to function as a tumor suppressor in murine hepatocytes, also participates in a molecular cascade that prevents calcifications in arterioles and medium-sized arteries. We further provide genetic evidence that this function of TIF1alpha is not exerted in hepatocytes. The sites of ectopic calcifications in mutant mice lacking TIF1alpha resemble those seen in mice carrying an activating mutation of the calcium sensor receptor (Casr) gene and, in TIF1alpha-deficient kidneys, Casr expression is increased together with that of many other vitamin D receptor (VDR) direct target genes, namely Car2, Cyp24a1, Trpv5, Trpv6, Calb1, S100g, Pthlh, and Spp1. Thus, our data indicate that TIF1alpha represses the VDR pathway in kidney and suggest that an up-regulation of Casr expression in this organ could account for ectopic calcifications generated upon TIF1alpha deficiency. Interestingly, the calcifying arteriopathy of TIF1alpha-null mutant mice shares features with the human age related Monckeberg's disease and, overall, the TIF1alpha-null mutant pathological phenotype supports the hypothesis that aging is promoted by increased activity of the vitamin D signaling pathway. PMID- 18287085 TI - Reanalyses of Gulf of Mexico fisheries data: landings can be misleading in assessments of fisheries and fisheries ecosystems. AB - We used two high profile articles as cases to demonstrate that use of fishery landings data can lead to faulty interpretations about the condition of fishery ecosystems. One case uses the mean trophic level index and its changes, and the other uses estimates of fishery collapses. In earlier analyses by other authors, marine ecosystems in the Gulf of Mexico (GOM) and U.S. Atlantic Ocean south of Chesapeake Bay were deemed to be severely overfished and the food webs badly deteriorated using these criteria. In our reanalyses, the low mean trophic level index for the GOM actually resulted from large catches of two groups of low trophic level species, menhaden and shrimp, and the mean trophic level was slowly increasing rather than decreasing. Commercial targeting and high landings of shrimps and menhaden, especially in the GOM, drove the index as previously calculated. Reanalyses of fishery collapses incorporating criteria that included targeting, variability in fishing effort, and market forces discovered many false cases of collapse based simply upon a decline of catches to 10% of previous maximum levels. Consequently, we suggest that the low mean trophic level index calculated in the earlier article for the GOM did not reflect the overall condition of the fishery ecosystem, and that the 10% rule for collapse should not be interpreted out of context in the GOM or elsewhere. In both cases, problems lay in the assumption that commercial landings data alone adequately reflect the fish populations and communities. PMID- 18287086 TI - Electrolyte concentration during haemodialysis and QT interval prolongation in uraemic patients. AB - AIMS: To assess the effect of different combinations of potassium and calcium concentrations on QT interval in the dialysis bath in uraemic patients. METHODS AND RESULTS: Sixteen haemodialysis (HD) patients underwent a 24 h Holter recording before and during HD sessions with six randomized combinations of electrolytes concentrations of the dialysis bath (K(+), 2 and 3 mmol/L; Ca(2+), 1.25, 1.5, and 1.75 mmol/L). The effect of different dialysis baths on QT interval was significant (P < 0.05). The longest mean QTc was observed with the lowest K(+) (2 mmol/L) and Ca(2+) concentrations (1.25 mmol/L), whereas the shortest mean QTc was observed with the highest K(+) (3 mmol/L) and Ca(2+) concentrations (1.75 mmol/L). QTc was >440 ms in 9 of 16 patients (56%) at the lowest Ca(2+) and K(+) concentrations, and in 3 of 16 patients (18%) at the highest electrolytes level. Changes in QTc during the HD sessions were inversely correlated with that in total Ca and Ca(2+) plasma concentrations (P < 0.0001). CONCLUSION: Changes in ventricular repolarization duration associated with HD largely depend on the concentrations of Ca(2+) and K(+) in the dialysis bath. These findings may have important implications for the choice of the electrolytes concentration of the dialysis bath during the HD session. PMID- 18287087 TI - Long-term follow-up after radiofrequency catheter ablation for atrial fibrillation. AB - AIMS: Data on long-term follow-up of patients who have undergone catheter ablation for atrial fibrillation (AF) are very limited. This report aimed at presenting clinical outcome and AF-free survival after pulmonary vein (PV) isolation over an extended (>3 years) follow-up period. METHODS AND RESULTS: Thirty-nine patients subjected to PV isolation for paroxysmal AF were followed-up for at least 3 years according to a strict protocol. Fourteen patients (35.8%) had one, 19 patients (50%) had two, and 6 patients (15.4%) had three ablation procedures. At end of follow-up (42.2 +/- 6.0 months), 17 patients (43.5%) were completely free of AF or other atrial arrhythmia, and 26 patients (66.6%) had symptomatic improvement. The long-term success rate was 21.4% for patients subjected to a single ablation procedure, 52.6% for patients subjected to two catheter ablation procedures, and 66.7% for patients who underwent three ablation procedures (P = 0.094). There was also a trend for patients who underwent a combination of different ablation procedures (ostial, antral, and/or circumferential) to have a higher AF-free survival when compared with patients subjected to the same procedure (P-value for log-rank test = 0.036). CONCLUSION: Catheter ablation does not eliminate paroxysmal AF in up to 56% of patients in the long term, despite the use of two or three ablation procedures in two-thirds of them. However, it confers symptomatic improvement in 67% of treated patients. PMID- 18287088 TI - Initial location of the RNA-dependent RNA polymerase in the bacteriophage Phi6 procapsid determined by cryo-electron microscopy. AB - The RNA-dependent RNA polymerases (RdRPs) of Cystoviridae bacteriophages, like those of eukaryotic viruses of the Reoviridae, function inside the inner capsid shell in both replication and transcription. In bacteriophage Phi6, this inner shell is first assembled as an icosahedral procapsid with recessed 5-fold vertices that subsequently undergoes major structural changes during maturation. The tripartite genome is packaged as single-stranded RNA molecules via channels on the 5-fold vertices, and transcripts probably exit the mature capsid by the same route. The RdRP (protein P2) is assembled within the procapsid, and it was thought that it should be located on the 5-fold axes near the RNA entry and exit channels. To determine the initial location of the RdRP inside the procapsid of bacteriophage Phi6, we performed cryo-electron microscopy of wild type and mutant procapsids and complemented these data with biochemical determinations of copy numbers. We observe ring-like densities on the 3-fold axes that are strong in a mutant that has approximately 10 copies of P2 per particle; faint in wild type, reflecting the lower copy number of approximately 3; and completely absent in a P2-null mutant. The dimensions and shapes of these densities match those of the known crystal structure of the P2 monomer. We propose that, during maturation, the P2 molecules rotate to occupy positions closer to adjacent 5-fold vertices where they conduct replication and transcription. PMID- 18287089 TI - Transforming growth factor-beta1 (TGFbeta1) stimulates connective tissue growth factor (CCN2/CTGF) expression in human gingival fibroblasts through a RhoA independent, Rac1/Cdc42-dependent mechanism: statins with forskolin block TGFbeta1-induced CCN2/CTGF expression. AB - Regulation of connective tissue growth factor (CCN2/CTGF) in gingival fibroblasts is unique and may provide therapeutic opportunities to treat oral fibrotic diseases. RhoA was previously implicated in mediating the expression of CCN2/CTGF. We now present evidence that Rho family GTPases Rac1 and Cdc42 are the principal mediators of the transforming growth factor-beta1 (TGFbeta1)-stimulated expression of CCN2/CTGF in primary human gingival fibroblasts. TGFbeta1 does not stimulate RhoA activation in gingival fibroblasts, and the overexpression of dominant-negative RhoA does not reduce CCN2/CTGF expression in response to TGFbeta1. In contrast, the overexpression of dominant-negative forms of Cdc42 or Rac1 results in a dramatic reduction of CCN2/CTGF protein levels. Lovastatin and a geranylgeranyltransferase inhibitor reduce the TGFbeta1-stimulated levels of CCN2/CTGF protein by approximately 75 and 100%, respectively. We previously demonstrated that JNK1 phosphorylation by TGFbeta1 is also critical for TGFbeta1 induced CCN2/CTGF expression, and forskolin partially reduces levels of phosphorylated JNK1. Inhibition of geranylgeranyltransferase has no effect on levels of JNK phosphorylation in response to TGFbeta1 suggesting Rho-GTPases act independently of JNK1. The combination of lovastatin and forskolin results in a greater inhibitory effect than each agent alone and reduces CCN2/CTGF mRNA and protein expression by greater than 90%. This novel combination has additive inhibitory effects on the TGFbeta1-stimulated expression of CCN2/CTGF in human gingival fibroblasts through the simultaneous disruption of Rho- and JNK1 mediated pathways, respectively. This combination of available therapeutic compounds may therefore be useful in designing treatment strategies for oral fibrotic conditions in which gingival CCN2/CTGF is elevated. PMID- 18287090 TI - Cdc28 and Cdc14 control stability of the anaphase-promoting complex inhibitor Acm1. AB - The anaphase-promoting complex (APC) regulates the eukaryotic cell cycle by targeting specific proteins for proteasomal degradation. Its activity must be strictly controlled to ensure proper cell cycle progression. The co-activator proteins Cdc20 and Cdh1 are required for APC activity and are important regulatory targets. Recently, budding yeast Acm1 was identified as a Cdh1 binding partner and APC(Cdh1) inhibitor. Acm1 disappears in late mitosis when APC(Cdh1) becomes active and contains conserved degron-like sequences common to APC substrates, suggesting it could be both an inhibitor and substrate. Surprisingly, we found that Acm1 proteolysis is independent of APC. A major determinant of Acm1 stability is phosphorylation at consensus cyclin-dependent kinase sites. Acm1 is a substrate of Cdc28 cyclin-dependent kinase and Cdc14 phosphatase both in vivo and in vitro. Mutation of Cdc28 phosphorylation sites or conditional inactivation of Cdc28 destabilizes Acm1. In contrast, inactivation of Cdc14 prevents Acm1 dephosphorylation and proteolysis. Cdc28 stabilizes Acm1 in part by promoting binding of the 14-3-3 proteins Bmh1 and Bmh2. We conclude that the opposing actions of Cdc28 and Cdc14 are primary factors limiting Acm1 to the interval from G(1)/S to late mitosis and are capable of establishing APC-independent expression patterns similar to APC substrates. PMID- 18287091 TI - Ikaros directly represses the notch target gene Hes1 in a leukemia T cell line: implications for CD4 regulation. AB - Ikaros and Notch1, two regulators of gene transcription, are critically important at many stages of T cell development. Deregulation of Ikaros and Notch activities cooperate to promote T cell leukemogenesis, providing evidence that they function in converging pathways in developing T cells. In this report, a mechanism for Ikaros:Notch cooperativity is described, revealing a non-redundant role for Ikaros in regulating expression of the Notch target gene Hes1 in a leukemia T cell line. We provide evidence that Ikaros directly represses Hes1 in concert with the transcriptional repressor, RBP-Jkappa, allowing for cross-talk between Notch and Ikaros that impacts regulation of CD4 expression. Taken together, these data describe a potential mechanism for Ikaros' function during T cell development and define Ikaros as an obligate repressor of Hes1. PMID- 18287092 TI - Human neutrophils convert the sebum-derived polyunsaturated fatty acid Sebaleic acid to a potent granulocyte chemoattractant. AB - Sebaleic acid (5,8-octadecadienoic acid) is the major polyunsaturated fatty acid in human sebum and skin surface lipids. The objective of the present study was to investigate the metabolism of this fatty acid by human neutrophils and to determine whether its metabolites are biologically active. Neutrophils converted sebaleic acid to four major products, which were identified by their chromatographic properties, UV absorbance, and mass spectra as 5-hydroxy-(6E,8Z) octadecadienoic acid (5-HODE), 5-oxo-(6E,8Z)-octadecadienoic acid (5-oxo-ODE), 5S,18-dihydroxy-(6E,8Z)-octadecadienoic acid, and 5-oxo-18-hydroxy-(6E,8Z) octadecadienoic acid. The identities of these metabolites were confirmed by comparison of their properties with those of authentic chemically synthesized standards. Both neutrophils and human keratinocytes converted 5-HODE to 5-oxo ODE. This reaction was stimulated in neutrophils by phorbol myristate acetate and in keratinocytes by oxidative stress (t-butyl-hydroperoxide). Both treatments dramatically elevated intracellular levels of NADP(+), the cofactor required by 5 hydroxyeicosanoid dehydrogenase. In keratinocytes, this was accompanied by a rapid increase in intracellular GSSG levels, consistent with the involvement of glutathione peroxidase. 5-Oxo-ODE stimulated calcium mobilization in human neutrophils and induced desensitization to 5-oxo-6,8,11,14-eicosatetraenoic acid but not leukotriene B(4), indicating that this effect was mediated by the OXE receptor. 5-Oxo-ODE and its 8-trans isomer were equipotent with 5-oxo-6,8,11,14 eicosatetraenoic acid in stimulating actin polymerization and chemotaxis in human neutrophils, whereas 5-HODE, 5-oxo-18-hydroxy-(6E,8Z)-octadecadienoic acid, and 5S,18-dihydroxy-(6E,8Z)-octadecadienoic acid were much less active. We conclude that neutrophil 5-lipoxygenase converts sebaleic acid to 5-HODE, which can be further metabolized to 5-oxo-ODE by 5-hydroxyeicosanoid dehydrogenase in neutrophils and keratinocytes. Because of its chemoattractant properties, sebum derived 5-oxo-ODE could be involved in neutrophil infiltration in inflammatory skin diseases. PMID- 18287093 TI - MEK signaling is required for phosphorylation of eIF2alpha following amino acid limitation of HepG2 human hepatoma cells. AB - The mammalian amino acid response (AAR) pathway is up-regulated by protein or amino acid depletion. This pathway involves detection of uncharged tRNA by the GCN2 kinase, phosphorylation of the translation initiation factor eIF2alpha (eukaryotic initiation factor 2alpha), and, through subsequent translational control, enhanced de novo synthesis of the transcription factor ATF4. The present studies demonstrate that inhibition of MEK activation in HepG2 human hepatoma cells by PD98059 or U0126 blocked the increased phosphorylation of eIF2alpha and ATF4 synthesis triggered by amino acid limitation, showing that the AAR requires activation of the MEK-ERK pathway. Inhibitors of the JNK or p38 MAPK pathways were ineffective. Consequently, inhibition of MEK activation blocked transcriptional induction of ATF4 target genes, but the induction was rescued by overexpression of ATF4 protein. Furthermore, the enhanced ERK phosphorylation following amino acid deprivation required GCN2 kinase activity and eIF2alpha phosphorylation. Inhibition of protein phosphatase 1 action on phospho-eIF2alpha by knockdown of GADD34 did not block the sensitivity to PD98059, suggesting that MEK functions to enhance GCN2-dependent eIF2alpha phosphorylation rather than suppressing dephosphorylation. Collectively, these results document a critical interdependence between the MEK-ERK MAPK signaling pathway and the amino acid stress-activated pathway. PMID- 18287094 TI - Suppression of interferon (IFN)-inducible genes and IFN-mediated functional responses in BCR-ABL-expressing cells. AB - The interferons (IFNs) are cytokines that play key roles in host defense against viral infections and immune surveillance against cancer. We report that BCR-ABL transformation of hematopoietic cells results in suppression of IFN-dependent responses, including transcription of IFN-inducible genes and generation of IFN mediated antiviral effects. BCR-ABL transformation suppresses expression of several IFN-regulated genes containing IFN-sensitive response element (ISRE) or GAS elements in their promoters, including Isg15, Irf1, Irf9, and Ifit2 (interferon-induced protein with tetratricopeptide repeats 2). Suppression of transcription of ISRE-containing genes is also seen in cells expressing various BCR-ABL kinase domain mutants, including T315I, H396P, Y253F, and E255K, but not kinase-defective BCR-ABL. Such effects are associated with impaired IFN-dependent phosphorylation of Stat1 on Tyr(701) and Stat3 on Tyr(705) and defective binding of Stat complexes to ISRE or GAS elements. Beyond suppression of Stat activities, BCR-ABL inhibits IFN-inducible phosphorylation/activation of the p38 MAPK, suggesting a dual mechanism by which this abnormal fusion protein blocks IFN transcriptional responses. The inhibitory activities of BCR-ABL ultimately result in impaired IFNalpha-mediated protection against encephalomyocarditis virus infection and reversal of IFN-dependent growth suppression. Altogether, our data provide evidence for a novel mechanism by which BCR-ABL impairs host defenses and promotes malignant transformation, involving dual suppression of IFN-activated signaling pathways. PMID- 18287095 TI - ISG15 inhibits Nedd4 ubiquitin E3 activity and enhances the innate antiviral response. AB - Interferons regulate diverse immune functions through the transcriptional activation of hundreds of genes involved in anti-viral responses. The interferon inducible ubiquitin-like protein ISG15 is expressed in cells in response to a variety of stress conditions like viral or bacterial infection and is present in its free form or is conjugated to cellular proteins. In addition, protein ubiquitination plays a regulatory role in the immune system. Many viruses modulate the ubiquitin (Ub) pathway to alter cellular signaling and the antiviral response. Ubiquitination of retroviral group-specific antigen precursors and matrix proteins of the Ebola, vesicular stomatitis, and rabies viruses by Nedd4 family HECT domain E3 ligases is an important step in facilitating viral release. We found that Nedd4 is negatively regulated by ISG15. Free ISG15 specifically bound to Nedd4 and blocked its interaction with Ub-E2 molecules, thus preventing further Ub transfer from E2 to E3. Furthermore, overexpression of ISG15 diminished the ability of Nedd4 to ubiquitinate viral matrix proteins and led to a decrease in the release of Ebola VP40 virus-like particles from the cells. These results point to a mechanistically novel function of ISG15 in the enhancement of the innate anti-viral response through specific inhibition of Nedd4 Ub-E3 activity. To our knowledge, this is the first example of a Ub-like protein with the ability to interfere with Ub-E2 and E3 interaction to inhibit protein ubiquitination. PMID- 18287096 TI - Unexpected complexity in the mechanisms that target assembly of the spectrin cytoskeleton. AB - The spectrin cytoskeleton assembles within discrete regions of the plasma membrane in a wide range of animal cell types. Although recent studies carried out in vertebrate systems indicate that spectrin assembly occurs indirectly through the adapter protein ankyrin, recent studies in Drosophila have established that spectrin can also assemble through a direct ankyrin-independent mechanism. Here we tested specific regions of the spectrin molecule for a role in polarized assembly and function. First, we tested mutant beta-spectrins lacking ankyrin binding activity and/or the COOH-terminal pleckstrin homology (PH) domain for their assembly competence in midgut, salivary gland, and larval brain. Remarkably, three different assembly mechanisms operate in these three cell types: 1) neither site was required for assembly in salivary gland; 2) only the PH domain was required in midgut copper cells; and 3) either one of the two sites was sufficient for spectrin assembly in larval brain. Further characterization of the PH domain revealed that it binds strongly to lipid mixtures containing phosphatidylinositol 4,5-bisphosphate (PIP(2)) but not phosphatidylinositol 3,4,5 trisphosphate. A K8Q mutation in the lipid binding region of the PH domain eliminated the PIP(2) interaction in vitro, yet the mutant protein retained full biological function in vivo. Reporter gene studies revealed that PIP(2) and the spectrin PH domain codistribute with one another in cells but not with authentic wild type alphabeta-spectrin. Thus, it appears that the PH domain imparts membrane targeting activity through a second mechanism that takes precedence over its PIP(2) binding activity. PMID- 18287097 TI - An amphipathic helical region of the N-terminal barrel of phospholipid transfer protein is critical for ABCA1-dependent cholesterol efflux. AB - Phospholipid lipid transfer protein (PLTP) mimics high-density lipoprotein apolipoproteins in removing cholesterol and phospholipids from cells through the ATP-binding cassette transporter A1 (ABCA1). Because amphipathic alpha-helices are the structural determinants for ABCA1 interactions, we examined the ability of synthetic peptides corresponding to helices in PLTP to remove cellular cholesterol by the ABCA1 pathway. Of the seven helices tested, only one containing PLTP residues 144-163 (p144), located at the tip of the N-terminal barrel, promoted ABCA1-dependent cholesterol efflux and stabilized ABCA1 protein. Mutating methionine 159 (Met-159) in this helix in PLTP to aspartate (M159D) or glutamate (M159E) nearly abolished the ability of PLTP to remove cellular cholesterol and dramatically reduced PLTP binding to phospholipid vesicles and its phospholipid transfer activity. These mutations impaired PLTP binding to ABCA1-generated lipid domains and PLTP-mediated stabilization of ABCA1 but increased PLTP binding to ABCA1. PLTP interactions with ABCA1 also mimicked apolipoproteins in activating Janus kinase 2; however, the M159D/E mutants were also able to activate this kinase. Structural analyses showed that the M159D/E mutations had only minor effects on PLTP conformation. These findings indicate that PLTP helix 144-163 is critical for removing lipid domains formed by ABCA1, stabilizing ABCA1 protein, interacting with phospholipids, and promoting phospholipid transfer. Direct interactions with ABCA1 and activation of signaling pathways likely involve other structural determinants of PLTP. PMID- 18287098 TI - A molecular switch for targeting between endoplasmic reticulum (ER) and mitochondria: conversion of a mitochondria-targeting element into an ER-targeting signal in DAKAP1. AB - dAKAP1 (AKAP121, S-AKAP84), a dual specificity PKA scaffold protein, exists in several forms designated as a, b, c, and d. Whether dAKAP1 targets to endoplasmic reticulum (ER) or mitochondria depends on the presence of the N-terminal 33 amino acids (N1), and these N-terminal variants are generated by either alternative splicing and/or differential initiation of translation. The mitochondrial targeting motif, which is localized between residues 49 and 63, is comprised of a hydrophobic helix followed by positive charges ( Ma, Y., and Taylor, S. (2002) J. Biol. Chem. 277, 27328-27336 ). dAKAP1 is located on the cytosolic surface of mitochondria outer membrane and both smooth and rough ER membrane. A single residue, Asp(31), within the first 33 residues of dAKAP1b is required for ER targeting. Asp(31), which functions as a separate motif from the mitochondrial targeting signal, converts the mitochondrial-targeting signal into a bipartite ER targeting signal, without destroying the mitochondria-targeting signal. Therefore dAKAP1 possesses a single targeting element capable of targeting to both mitochondria and ER, with the ER signal overlapping the mitochondria signal. The specificity of ER or mitochondria targeting is determined and switched by the availability of the negatively charged residue, Asp(31). PMID- 18287099 TI - Phospholamban oligomerization, quaternary structure, and sarco(endo)plasmic reticulum calcium ATPase binding measured by fluorescence resonance energy transfer in living cells. AB - Phospholamban (PLB) oligomerization, quaternary structure, and sarco(endo)plasmic reticulum calcium ATPase (SERCA) binding were quantified by fluorescence resonance energy transfer (FRET) in an intact cellular environment. FRET between cyan fluorescent protein-PLB and yellow fluorescent protein-PLB in AAV-293 cells showed hyperbolic dependence on protein concentration, with a maximum efficiency of 45.1 +/- 1.3%. The observed FRET corresponds to a probe separation distance of 58.7 +/- 0.5A(,) according to a computational model of intrapentameric FRET. This is consistent with models of the PLB pentamer in which cytoplasmic domains fan out from the central bundle of transmembrane helices. An I40A mutation of PLB did not alter pentamer conformation but increased the concentration of half-maximal FRET (K(D)) by >4-fold. This is consistent with the previous observation that this putatively monomeric mutant still oligomerizes in intact membranes but forms more dynamic pentamers than wild type PLB. PLB association with SERCA, measured by FRET between cyan fluorescent protein-SERCA and yellow fluorescent protein PLB, was increased by the I40A mutation without any detectable change in probe separation distance. The data indicate that the regulatory complex conformation is not altered by the I40A mutation. A naturally occurring human mutation (L39Stop) greatly reduced PLB oligomerization and SERCA binding and caused mislocalization of PLB to the cytoplasm and nucleus. Overall, the data suggest that the PLB pentamer adopts a "pinwheel" shape in cell membranes, as opposed to a more compact "bellflower" conformation. I40A mutation decreases oligomerization and increases PLB binding to SERCA. Truncation of the transmembrane domain by L39Stop mutation prevents anchoring of the protein in the membrane, greatly reducing PLB binding to itself or its regulatory target, SERCA. PMID- 18287100 TI - A novel function for hydroxyproline oxidase in apoptosis through generation of reactive oxygen species. AB - Proline and hydroxyproline are metabolized by distinct pathways. Proline is important for protein synthesis, as a source of glutamate, arginine, and tricarboxylic acid cycle intermediates, and for participating in a metabolic cycle that shuttles redox equivalents between mitochondria and cytosol. Hydroxyproline, in contrast, is not reutilized for protein synthesis. The first steps in the degradation of proline and hydroxyproline are catalyzed by proline oxidase (POX) and hydroxyproline oxidase (OH-POX), respectively. Because it is well documented that POX is induced by p53 and plays a role in apoptosis, we considered whether OH-POX also participates in the response to cytotoxic stress. In LoVo and RKO cells, which respond to adriamycin with a p53-mediated induction of POX and generation of reactive oxygen species, we found that adriamycin also induced OH-POX gene expression and markedly increased OH-POX catalytic activity, and this increase in activity was not observed in the cell lines HT29 and HCT15, which do not have a functional p53. We also observed an increase in reactive oxygen species generation and activation of caspase-9 with adriamycin in a hydroxyproline-dependent manner. Therefore, we hypothesize that OH-POX plays a role analogous to POX in growth regulation, ROS generation, and activation of the apoptotic cascade. PMID- 18287101 TI - The ATPase cycle of the mitochondrial Hsp90 analog Trap1. AB - Hsp90 is an ATP-dependent molecular chaperone whose mechanism is not yet understood in detail. Here, we present the first ATPase cycle for the mitochondrial member of the Hsp90 family called Trap1 (tumor necrosis factor receptor-associated protein 1). Using biochemical, thermodynamic, and rapid kinetic methods we dissected the kinetics of the nucleotide-regulated rearrangements between the open and the closed conformations. Surprisingly, upon ATP binding, Trap1 shifts predominantly to the closed conformation (70%), but, unlike cytosolic Hsp90 from yeast, this process is rather slow at 0.076 s(-1). Because reopening (0.034 s(-1)) is about ten times faster than hydrolysis (k(hyd) = 0.0039 s(-1)), which is the rate-limiting step, Trap1 is not able to commit ATP to hydrolysis. The proposed ATPase cycle was further scrutinized by a global fitting procedure that utilizes all relevant experimental data simultaneously. This analysis corroborates our model of a two-step binding mechanism of ATP followed by irreversible ATP hydrolysis and a one-step product (ADP) release. PMID- 18287102 TI - Crystal structure of the ligand-bound glucagon-like peptide-1 receptor extracellular domain. AB - The glucagon-like peptide-1 receptor (GLP-1R) belongs to Family B1 of the seven transmembrane G protein-coupled receptors, and its natural agonist ligand is the peptide hormone glucagon-like peptide-1 (GLP-1). GLP-1 is involved in glucose homeostasis, and activation of GLP-1R in the plasma membrane of pancreatic beta cells potentiates glucose-dependent insulin secretion. The N-terminal extracellular domain (nGLP-1R) is an important ligand binding domain that binds GLP-1 and the homologous peptide Exendin-4 with differential affinity. Exendin-4 has a C-terminal extension of nine amino acid residues known as the "Trp cage", which is absent in GLP-1. The Trp cage was believed to interact with nGLP-1R and thereby explain the superior affinity of Exendin-4. However, the molecular details that govern ligand binding and specificity of nGLP-1R remain undefined. Here we report the crystal structure of human nGLP-1R in complex with the antagonist Exendin-4(9-39) solved by the multiwavelength anomalous dispersion method to 2.2A resolution. The structure reveals that Exendin-4(9-39) is an amphipathic alpha-helix forming both hydrophobic and hydrophilic interactions with nGLP-1R. The Trp cage of Exendin-4 is not involved in binding to nGLP-1R. The hydrophobic binding site of nGLP-1R is defined by discontinuous segments including primarily a well defined alpha-helix in the N terminus of nGLP-1R and a loop between two antiparallel beta-strands. The structure provides for the first time detailed molecular insight into ligand binding of the human GLP-1 receptor, an established target for treatment of type 2 diabetes. PMID- 18287103 TI - Public health, the environment and the 21st century: a wider field of vision. PMID- 18287104 TI - Alcohol-related adverse consequences: cross-cultural variations in attribution process among young adults. AB - BACKGROUND: Social norms around what is culturally accepted in terms of alcohol consumption and drunken comportment appear important regarding the acceptance of alcohol-related adverse consequences; however, investigations often neglect to consider differences in terms of attribution. This study aims at assessing cross cultural differences in the reporting of alcohol-related adverse consequences. It also considers differences across consequences that might explain which type of consequences (mainly acute or mainly chronic) are most affected by an attribution process. METHODS: Conditional regression models were estimated based on data from eight European countries participating in the Gender, Alcohol and Culture--An International Study (GENACIS) project. Cases were matched to controls based on usual drinking patterns in order to control for average volume of alcohol and frequency of 'risky single occasion drinking' (RSOD). RESULTS: Differences among the patterns of associations between countries and consequences were evident. The distinction between Nordic and other European countries was persistent. A higher variability of associations was observed for some consequences, namely the mainly acute instances. Finally, the Isle of Man and Switzerland showed specific trends with associations across consequences. CONCLUSION: Reporting of alcohol-related adverse consequences seemed strongly affected by cultural norms. The latter may be exemplified by viewing drinking as 'time-out' behaviour. Respondents in countries with a stereotypical history of being 'dry' or with a stereotyped 'binge' drinking culture were more likely to attribute consequences to their alcohol consumption than people in 'wet' countries. This was particularly true for consequences that related to episodic 'time-out' heavy drinking. PMID- 18287105 TI - Photodynamic ablation of a selected rat embryo: a model for the treatment of extrauterine pregnancy. AB - BACKGROUND: To test the feasibility of photodynamic therapy (PDT)-based ablation of rat embryos as a model for PDT of extrauterine pregnancy (EUP) in humans. METHODS: A controlled pre-clinical study. Selected rat embryos [one per litter, n = 30, embryonic day 14 (E14)] were subjected to placental injection of a Palladium-bacteriochlorophyll derivative and illuminated to achieve selective photo-ablation. Histopathology studies were performed 48 h after treatment (E16). Parturition (E21) and breeding (approximately 12 weeks) after treatment were also evaluated. RESULTS: Using direct placental injection, nearly 80% of the treated rat embryos were selectively photo-ablated, leaving the remaining litter unharmed to achieve normal parturition. Treated animals retained fertility and normally implanted in both treated and untreated uterine horns attesting to the confined toxicity inherent to this approach. CONCLUSIONS: Although requiring respective adaptation to clinical application in terms of treatment protocols and designated hardware, photodynamic interventions using novel bacteriochlorophyll-based photosensitizers may prove applicable to treatment of EUP, as well as other gynecological pathologies and malignancies in a safe, minimally invasive manner. PMID- 18287106 TI - The presence of heparan sulfate in the mammalian oocyte provides a clue to human sperm nuclear decondensation in vivo. AB - BACKGROUND: Previous results from our laboratory have led us to propose heparan sulfate (HS) as a putative protamine acceptor during human sperm decondensation in vivo. The aim of this paper was to investigate the presence of glycosaminoglycans in the mammalian oocyte in an effort to better support this contention. METHODS: Two experimental approaches are used: oocyte labeling to identify the presence of HS and analysis of sperm decondensing ability of fresh oocytes in the presence or absence of specific glycosidases. RESULTS: Staining of mouse zona-intact oocytes with the fluorescent cationic dye, Rubipy, at pH 1.5 allowed for the detection of sulfate residues in the ooplasm by confocal microscopy. HS was detected in the ooplasm by immunocytochemistry. A sperm decondensation microassay using heparin and glutathione was successfully developed. The same level of sperm decondensation could be attained when heparin was replaced by mouse zona-free oocytes. Addition of heparinase to the oocyte/glutathione mixture significantly reduced sperm decondensation (P = 0.0159), while there was no effect following addition of either chondroitinase ABC or hyaluronidase. CONCLUSIONS: The results presented in this paper demonstrate for the first time that HS is present in the mammalian oocyte and show that HS is necessary for fresh oocytes to express their sperm decondensing ability in vitro. PMID- 18287107 TI - Sleep difficulties in infants at risk for developmental delays: a longitudinal study. AB - OBJECTIVES: We compared the sleep of infants at risk for neuromotor delays to that of infants without such risks, and examined the predictive validity of risk indicators to the development of sleep problems. METHODS: Conveniently recruited infants (n = 142) were assessed for neuromotor achievements and sleep behaviors at 4-6 months and 10-12 months of age. Assessment tools were the Harris Infant Neuromotor Test and Morrell's Infant Sleep Questionnaire. Based on a cumulative risk index, three groups were defined: higher risk (n = 28), lower risk (n = 42), and no risk (n = 72). RESULTS: At both ages, the sleep scores were similar among the groups. In the no risk and lower risk group, sleep difficulties decreased with age, while for infants in the higher risk group, more difficulties were reported over time. Overall, the neuromotor attainments were not related to sleep fragmentation or settling difficulties. CONCLUSIONS: In a diverse sample of infants, with and without risks for developmental delays, overall, sleep patterns were similar. It appears that the neuromotor achievements are not associated with sleep-wake regulation, as measured by caregivers' report. PMID- 18287108 TI - An integration of parents' and best friends' smoking, smoking-specific cognitions, and nicotine dependence in relation to readiness to quit smoking: a comparison between adolescents with and without asthma. AB - OBJECTIVE: To study the impact of parents' and best friends' smoking, nicotine dependence, and craving on smoking-specific cognitions, and readiness to quit in adolescents with and without asthma. METHODS: Structural equation analyses were applied to data from a sample of 1,120 daily smoking adolescents, 83 of whom had asthma. RESULTS: Adolescents with asthma felt more ready to quit, and cognitions were more strongly related to readiness to quit among adolescents with asthma than among adolescents without asthma. Moreover, best friends' smoking seemed more relevant to the cognitions of adolescents with asthma. Nicotine dependence and craving were strongly related to cognitions, and to readiness to quit in both groups. The relation between craving and readiness to quit, however, was stronger among participants with asthma. CONCLUSIONS: Reduction of nicotine dependence and craving is essential for both groups. Youth with asthma may benefit even more from cognitive-based cessation services than healthy youth. The finding that adolescents with asthma are relatively more ready to quit, and that their cognitions are more easily affected can be turned into advantages in asthma specific cessation services. PMID- 18287109 TI - Assessment of parental psychological stress in pediatric cancer: a review. AB - OBJECTIVES: We present an overview of the literature between 1997 and 2007 on parental stress reactions following the diagnosis of childhood cancer and we evaluate methodological strengths and weaknesses of the studies. METHODS: PubMed, PsychInfo, and Cinahl databases were used. Sixty-seven were included in the review. RESULTS: The conceptualization of parental stress and timing of assessment varies considerably between the studies, which makes comparison difficult. Most emotional stress reactions are seen around the time of diagnosis, with mothers reporting more symptoms than fathers. As a group, parents seem relatively resilient, although a subset of parents reports continuing stress even up to 5 years or more postdiagnosis. CONCLUSIONS: The authors recommend clear definitions of parental stress, fixed points in time to assess parental stress, and an approach that highlights both parental strengths and weaknesses. Improved assessment can contribute to tailoring psychological care to those parents most in need. PMID- 18287110 TI - Godoy & Godoy technique of cervical stimulation in the reduction of edema of the face after cancer treatment. PMID- 18287111 TI - Repeat thrombolysis or conservative therapy vs. rescue percutaneous coronary intervention for failed thrombolysis: systematic review and meta-analysis. AB - BACKGROUND: Despite proven advantages of primary percutaneous coronary intervention (PCI), thrombolysis remains the first line treatment for ST elevation myocardial infarction (STEMI) worldwide. Management of patients with failed thrombolysis is still debated, and data from existing randomized controlled trials are conflicting. AIM: To compare the risk/benefit profile of repeat thrombolysis (RT) vs. rescue PCI in patients with failed thrombolysis. METHODS: Search of BioMedCentral, CENTRAL, mRCT and PubMed for randomized controlled trials comparing rescue PCI vs. conservative therapy and/or RT vs. conservative therapy. Outcomes of interest assessed by adjusted indirect meta analysis: major adverse events (MAE, defined as the composite of overall mortality and re-infarction), stroke, congestive heart failure (CHF), major bleeds (MB), and minor bleeds. Overall mortality and re-infarction have been also analysed individually. RESULTS: Eight trials were included (1318 patients). Follow-up ranged from 'in-hospital' to 6 months. No significant difference was found for the risk of MAE [OR 0.93(0.26-3.35), P = 0.4], overall mortality [OR 1.01(0.52-1.95), P = 0.15], stroke [OR 5.03(0.64-39.1), P = 0.58] and CHF [OR 0.74(0.28-1.96), P = 0.6]. Compared with conservative therapy, rescue PCI was associated with a 70% reduction in the risk of re-infarction [OR 0.32(0.14-0.74), P = 0.008], number needed to treat 17. No difference in terms of MB was found [OR 0.5(0.1-2.5), P = 0.09], while a greater risk of minor bleeds was observed with rescue PCI [OR 2.48(1.08-5.7), P = 0.04], number needed to harm 50. CONCLUSION: Although the observed benefit is modest, these data support the use of PCI after failed thrombolysis. PMID- 18287112 TI - A pilot study of combination anti-cytokine and anti-lymphocyte biological therapy in rheumatoid arthritis. AB - BACKGROUND: Immunological tolerance in humans using anti-T-cell monoclonal antibodies (mAbs) may be hampered by a pro-inflammatory microenvironment. All clinical trials of such therapies in rheumatoid arthritis (RA), however, have selected patients with active disease at baseline. Concurrent neutralization of inflammation with a TNFalpha antagonist should maximize the potential of anti-T cell mAbs to induce tolerance in RA. AIM: To evaluate the safety of combining a TNFalpha antagonist and CD4 mAb in RA. DESIGN: An iterative pilot study focused on the safety of such combination therapy. METHODS: Eight poor prognosis, seropositive RA patients were treated with combined CD4 and TNFalpha blockade. Prolonged CD4 blockade was achieved with a humanized mAb, and TNFalpha blockade with a p55 TNF receptor fusion protein. RESULTS: There was a low incidence of classical first-dose reactions to the CD4 mAb, possibly reflecting concomitant TNFalpha blockade. An unusual anaphylactoid reaction was seen, however, and one patient developed a probable allergic reaction after several infusions. Skin rashes were common, as previously reported with CD4 mAb monotherapy. No serious infections were documented during follow-up, despite CD4+ lymphopenia in some patients. Most patients appeared to demonstrate improved RA disease control after the study. After 17-49 months after therapy, one patient was in remission, one remained off disease modifying anti-rheumatic drugs and five had stable disease, three on previously ineffective doses of methotrexate. CONCLUSION: We report, for the first time in man, immunotherapy with a combination of an anti-cytokine and an anti-T-cell reagent. We witnessed an unusual first-dose reaction but there were no significant infectious complications. PMID- 18287113 TI - Markers of exposure to spotted fever rickettsiae in patients with chronic illness, including fatigue, in two Australian populations. AB - BACKGROUND: Some investigators believe that a proportion of chronically unwell patients, many with fatigue, have an underlying rickettsial disease. AIM: To investigate the prevalence of markers of rickettsial infection in patients with chronic illnesses. DESIGN: Observational study. METHODS: A 526 patient cohort with chronic illnesses from Melbourne, Australia and 400 control patients from Newcastle, Australia were assessed using serology, culture and PCR for the detection of rickettsiae. Rickettsial serology was performed on another cohort of 581 chronically unwell patients (and 34 non-fatigued patients from the same practice) from Adelaide, Australia. RESULTS: Of the Melbourne patient cohort, 14/526 (3%) were real-time PCR positive for rickettsial DNA compared to none of the 400 control patients (P < 0.001). Of these 14 patients, Rickettsia honei strain 'marmionii' was detected in 5 and isolated from 2. Rickettsaemia was seasonal, with more in winter (8/145; P < 0.03) and less in spring (0/143; P < 0.03). Positive rickettsial serology titres of >or=1:256 were seen in 206 (39%) patients. Of the Adelaide patient cohort, 238/581 (41%) had positive rickettsial antibodies titres. Of the 34 control sera, 5 (15%) were serologically positive (P < 0.002). Both Melbourne and Adelaide patient cohorts had significantly higher seropositivity than the Newcastle control cohort (3/399; P < 0.0001). CONCLUSION: In patients with chronic illness, rickettsial DNA in peripheral blood and/or rickettsial seropositivity may represent exposure to rickettsiae or underlying rickettsial diseases. It is not known whether the presence of rickettsiae is causally related to the patients' chronic illnesses, or reactivation of a latent rickettsial infection. PMID- 18287114 TI - The U.S. economy and changes in health insurance coverage, 2000-2006. AB - The number of uninsured Americans increased by 3.4 million between 2004 and 2006, despite improving economic conditions. In the first four years of the decade, during a period of economic recession, the number increased by 6.0 million. The dominant factor in both periods was a decline in employer-sponsored insurance coverage. Although the recent decline was less than that experienced from 2000 to 2004, growth in public coverage was small, and the number of uninsured people increased by 1.0 million children and 2.4 million adults. Employer coverage declined most for self-employed or small-firm workers, in the South, and among noncitizens. PMID- 18287115 TI - PROMALS3D: a tool for multiple protein sequence and structure alignments. AB - Although multiple sequence alignments (MSAs) are essential for a wide range of applications from structure modeling to prediction of functional sites, construction of accurate MSAs for distantly related proteins remains a largely unsolved problem. The rapidly increasing database of spatial structures is a valuable source to improve alignment quality. We explore the use of 3D structural information to guide sequence alignments constructed by our MSA program PROMALS. The resulting tool, PROMALS3D, automatically identifies homologs with known 3D structures for the input sequences, derives structural constraints through structure-based alignments and combines them with sequence constraints to construct consistency-based multiple sequence alignments. The output is a consensus alignment that brings together sequence and structural information about input proteins and their homologs. PROMALS3D can also align sequences of multiple input structures, with the output representing a multiple structure based alignment refined in combination with sequence constraints. The advantage of PROMALS3D is that it gives researchers an easy way to produce high-quality alignments consistent with both sequences and structures of proteins. PROMALS3D outperforms a number of existing methods for constructing multiple sequence or structural alignments using both reference-dependent and reference-independent evaluation methods. PMID- 18287116 TI - Empirical comparison of ab initio repeat finding programs. AB - Identification of dispersed repetitive elements can be difficult, especially when elements share little or no homology with previously described repeats. Consequently, a growing number of computational tools have been designed to identify repetitive elements in an ab initio manner, i.e. without using prior sequence data. Here we present the results of side-by-side evaluations of six of the most widely used ab initio repeat finding programs. Using sequence from rice chromosome 12, tools were compared with regard to time requirements, ability to find known repeats, utility in identifying potential novel repeats, number and types of repeat elements recognized and compactness of family descriptions. The study reveals profound differences in the utility of the tools with some identifying virtually their entire substrate as repetitive, others making reasonable estimates of repetition, and some missing almost all repeats. Of note, even when tools recognized similar numbers of repeats they often showed marked differences in the nature and number of repeat families identified. Within the context of this comparative study, ReAS and RepeatScout showed the most promise in analysis of sequence reads and assembled genomic regions, respectively. Our results should help biologists identify the program(s), if any, that is best suited for their needs. PMID- 18287117 TI - Magnitude and direction of DNA bending induced by screw-axis orientation: influence of sequence, mismatches and abasic sites. AB - DNA-bending flexibility is central for its many biological functions. A new bending restraining method for use in molecular mechanics calculations and molecular dynamics simulations was developed. It is based on an average screw rotation axis definition for DNA segments and allows inducing continuous and smooth bending deformations of a DNA oligonucleotide. In addition to controlling the magnitude of induced bending it is also possible to control the bending direction so that the calculation of a complete (2-dimensional) directional DNA bending map is now possible. The method was applied to several DNA oligonucleotides including A(adenine)-tract containing sequences known to form stable bent structures and to DNA containing mismatches or an abasic site. In case of G:A and C:C mismatches a greater variety of conformations bent in various directions compared to regular B-DNA was found. For comparison, a molecular dynamics implementation of the approach was also applied to calculate the free energy change associated with bending of A-tract containing DNA, including deformations significantly beyond the optimal curvature. Good agreement with available experimental data was obtained offering an atomic level explanation for stable bending of A-tract containing DNA molecules. The DNA-bending persistence length estimated from the explicit solvent simulations is also in good agreement with experiment whereas the adiabatic mapping calculations with a GB solvent model predict a bending rigidity roughly two times larger. PMID- 18287118 TI - Surgical repair of aortico-left ventricular tunnel arising from the left aortic sinus. AB - Aortico-left ventricular tunnel is a rare congenital cardiac defect, which bypasses the aortic valve via the paravalvar connection from the left ventricle to the aorta. In most of the cases, the tunnel arises from the right aortic sinus. We herein report a case of aortico-left ventricular tunnel, of which the aortic orifice was arising from the left aortic sinus, requiring special attention for avoiding left coronary artery injury at the time of surgical repair. PMID- 18287119 TI - Does video-assisted thoracoscopic pleurectomy result in better outcomes than open pleurectomy for primary spontaneous pneumothorax? AB - The question addressed by a best evidence topic approach using a structured protocol was whether pleurectomy using video-assisted thoracoscopic surgery (VATS) resulted in better outcomes than open pleurectomy for primary spontaneous pneumothorax. Altogether 45 relevant papers were identified of which nine papers represented the best evidence to answer the question. The author, journal, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses were tabulated. We conclude that VATS pleurectomy has been shown to be comparable to open pleurectomy in the treatment of spontaneous pneumothorax, with a meta-analysis and several RCTs showing reductions in length of hospital stay and analgesic requirements. Postoperative pulmonary dysfunction has also been shown to be reduced after VATS pleurectomy in two RCTs, although a third study found no significant difference. A concern may be a four-fold increase in the recurrence of pneumothorax following VATS pleurectomy as compared to open pleurectomy reported in a recent meta-analysis of four randomised and 25 non-randomised studies performed in 2007 and published in the Lancet, although a second meta-analysis of only the randomised trials did not show this difference. PMID- 18287121 TI - John Ruskin's relapsing encephalopathy. AB - John Ruskin (1819-1900) is chiefly remembered for his works on painting and architecture, and for his powerful and original prose style. In middle age, he suffered recurring episodes of delirium with visual hallucinations and delusions. At about the same time, his writing developed a disjointed polemical character, with cryptic and intemperate elements that disorientated some readers. The nature of Ruskin's 'madness' is a key to understanding his later writing career but the psychiatric explanations given by many of his literary biographers seem unsatisfactory. Ruskin left numerous clues about the illness in his diaries, correspondence and publications. It is likely that he had a relapsing-progressive neurological disorder with neuropsychiatric manifestations. It could have been a fluctuating metabolic or immunological encephalopathy, but the diagnosis that best fits the time course of his illness and the prior history of mood disorder and of migraine with aura is Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL). Whatever the pathology, its first effects on frontal lobe function may have actually enhanced Ruskin's creative energy for a long time before stepwise cognitive impairment degraded his ability to write. PMID- 18287122 TI - Music listening enhances cognitive recovery and mood after middle cerebral artery stroke. AB - We know from animal studies that a stimulating and enriched environment can enhance recovery after stroke, but little is known about the effects of an enriched sound environment on recovery from neural damage in humans. In humans, music listening activates a wide-spread bilateral network of brain regions related to attention, semantic processing, memory, motor functions, and emotional processing. Music exposure also enhances emotional and cognitive functioning in healthy subjects and in various clinical patient groups. The potential role of music in neurological rehabilitation, however, has not been systematically investigated. This single-blind, randomized, and controlled trial was designed to determine whether everyday music listening can facilitate the recovery of cognitive functions and mood after stroke. In the acute recovery phase, 60 patients with a left or right hemisphere middle cerebral artery (MCA) stroke were randomly assigned to a music group, a language group, or a control group. During the following two months, the music and language groups listened daily to self selected music or audio books, respectively, while the control group received no listening material. In addition, all patients received standard medical care and rehabilitation. All patients underwent an extensive neuropsychological assessment, which included a wide range of cognitive tests as well as mood and quality of life questionnaires, one week (baseline), 3 months, and 6 months after the stroke. Fifty-four patients completed the study. Results showed that recovery in the domains of verbal memory and focused attention improved significantly more in the music group than in the language and control groups. The music group also experienced less depressed and confused mood than the control group. These findings demonstrate for the first time that music listening during the early post-stroke stage can enhance cognitive recovery and prevent negative mood. The neural mechanisms potentially underlying these effects are discussed. PMID- 18287123 TI - Coverage and yield of entry and follow-up screening for tuberculosis among new immigrants. AB - The aim of the present study was to determine the effectiveness of entry screening for tuberculosis and biannual follow-up screening among new immigrants in The Netherlands. To achieve this, the present authors analysed screening, prevalence and incidence data of 68,122 immigrants, who were followed for 29 months. Patients diagnosed within 5 months and 6-29 months after entry screening were considered to be detected at entry and during the follow-up period, respectively. Coverage of the second to fifth screening rounds was 59, 46, 36 and 34%, respectively. Yield of entry screening was 119 per 100,000 individuals, and prevalence at entry was 131 per 100,000. Average yield of follow-up screening was highest among immigrants with abnormalities on chest radiography (CXR) at entry (902 per 100,000 individuals). When excluding these, yield of follow-up screening was 9, 37 and 97 per 100,000 screenings for immigrants from countries with tuberculosis incidences of <100, 100-200 and >200 per 100,000, respectively. The incidence during follow-up in individuals with a normal CXR was 11, 58 and 145 per 100,000 person-yrs follow-up in these groups. The proportion of cases detected through screening declined per screening round from 91 to 31%. Yield of entry screening was high. Overall coverage and yield of follow-up screening was low. Follow-up screening of immigrants with a normal chest radiograph from countries with an incidence of <200 per 100,000 individuals was therefore discontinued. PMID- 18287124 TI - Adrenal function in patients with community-acquired pneumonia. AB - Adrenal insufficiency is believed to occur frequently in severe sepsis and septic shock. The aim of the present study was to determine whether adrenal function is also related to the severity of community-acquired pneumonia (CAP). In total, 64 Japanese patients with CAP were consecutively enrolled in the present study, which was carried out during 2005-2006. Serum adrenocorticotropic hormone (ACTH) and cortisol were measured in each subject, as was the response of cortisol secretion when 250 mug of cosyntropin was administered. Analyses were performed comparing these values with the score calculated according to the Pneumonia Patient Outcomes Research Team (PORT) cohort study, the number of in-hospital deaths and the length of hospital stay. As the PORT score increased, serum ACTH and cortisol also increased, while the response of cortisol secretion to the administration of cosyntropin decreased. In the analysis by receiver operating characteristic curves, adrenal dysfunction was related significantly to both the number of in-hospital deaths and the length of hospital stay. Adrenal dysfunction was shown to correlate with the Pneumonia Patient Outcomes Research Team score and the clinical outcomes, while adrenal insufficiency defined by the cosyntropin stimulation test was rare in the present study. PMID- 18287125 TI - Children at high altitude have less nocturnal periodic breathing than adults. AB - Although children commonly travel to high altitudes, their respiratory adaptation to hypoxia remains elusive. Therefore, in the present study respiratory inductive plethysmography, pulse oximetry (S(p,O(2))) and end-tidal CO(2) tension (P(ET,CO(2))) were recorded in 20 pre-pubertal children (aged 9-12 yrs) and their fathers during 1 night in Zurich (490 m) and 2 nights at the Swiss Jungfrau-Joch research station (3,450 m) following ascent by train within <3 h. In children, mean+/-sd nocturnal S(p,O(2)) fell from 98+/-1% at 490 m to 85+/-4 and 86+/-4% at 3,450 m (nights 1 and 2, respectively); P(ET,CO(2)) decreased significantly from 37+/-6 to 32+/-3 and 33+/-4 mmHg (3,450 versus 490 m). In adults, changes in nocturnal S(p,O(2)) and P(ET,CO(2)) at 3,450 m were similar to those in children. Children spent less time in periodic breathing at 3,450 m during night 1 and 2 (8+/-11 and 9+/-13%, respectively) than adults (34+/-24 and 22+/-17%, respectively), and their apnoea threshold for CO(2) was lower compared with adults (27+/-2 and 30+/-2 mmHg, respectively, both nights). S(p,O(2)), P(ET,CO(2)) and time in periodic breathing at altitude were not correlated between children and their fathers. In conclusion, children revealed a similarly reduced nocturnal O(2) saturation and associated hyperventilation at high altitude as adults but their breathing pattern was more stable, possibly related to a lower apnoea threshold for CO(2). PMID- 18287126 TI - Adenosine 5'-monophosphate in asthma: gas exchange and sputum cellular responses. AB - Adenosine 5'-monophosphate (AMP) bronchoprovocation reproduces the lung function abnormalities that occur spontaneously during acute asthma and detects peripheral airway inflammation better than direct bronchoconstrictive agents. Pulmonary gas exchange disturbances may reflect changes in small airways related to airway inflammation rather than bronchoconstriction alone. The present authors investigated whether AMP induced a greater imbalance in the ventilation/perfusion ratio than methacholine (MCh), at an equivalent degree of bronchoconstriction, with and without salbutamol pre-medication. In total, 36 asthmatics were studied in three randomised, double-blind, crossover studies: 1) before and after AMP or MCh; 2) before and 30 min after salbutamol or placebo, followed by AMP; or 3) MCh challenge. Sputum was collected before and 4 h post-challenge. Compared with MCh, AMP provoked similar pulmonary gas exchange abnormalities at an equivalent degree of intense bronchoconstriction (forced expiratory volume in one second decrease of 28-44%). While salbutamol blocked AMP- or MCh-induced bronchoconstriction, arterial oxygen tension (P(a,O(2))) and alveolar-arterial oxygen tension difference (P(A-a,O(2))) disturbances induced by AMP and MCh were only partially blocked (P(a,O(2)) by 46 and 42%, respectively; P(A-a,O(2)) by 58 and 57%, respectively). Compared with MCh, AMP increased the percentage of neutrophils (mean+/-se increased from 28+/-4% to 38+/-4%), but this increase did not occur after salbutamol pre-treatment. Both adenosine 5'-monophosphate and methacholine induced similar peripheral airway dysfunction. The fully inhibited adenosine 5' monophosphate-induced neutrophilia with residual hypoxaemia observed after salbutamol treatment is probably related to beta(2)-agonists acting on both bronchial and pulmonary circulation. PMID- 18287127 TI - High levels of Epstein-Barr virus in COPD. AB - Latent viral infection has been implicated in the pathophysiology of chronic obstructive pulmonary disease (COPD). Epstein-Barr virus (EBV) is known to be important in pulmonary fibrosis. The current authors hypothesised that EBV is associated with the pathogenesis of COPD. Sputum samples were collected from patients both during exacerbations of COPD and when stable. A control group of smokers who did not have airways obstruction also had their sputum examined. The presence of EBV DNA was established and quantified using a real-time nucleic acid amplification assay. A total of 136 patients with COPD were recruited during an acute exacerbation and a total of 68 when stable. EBV was detected in 65 (48%) exacerbation cases and 31 (46%) stable patients. In the comparison group of 16 nonobstructed smokers, EBV was demonstrated in only one (6%) case. Risk of COPD in patients with EBV and who are smokers confers an odds ratio of 12.6. Epstein Barr virus DNA is more frequently identified in the respiratory tract of chronic obstructive pulmonary disease patients in comparison with unaffected smokers. It is present both during exacerbation and when stable, suggesting that infection is persistent. Smokers who do not develop chronic obstructive pulmonary disease rarely have Epstein-Barr virus in their sputum. This finding may be of importance in the pathogenesis of chronic obstructive pulmonary disease. PMID- 18287128 TI - How to quantify coughing: correlations with quality of life in chronic cough. AB - Different methods are used for quantifying coughing in sound recordings, but as yet no method has been shown to be more valid than any other. In the present study, the relationships between three different units of cough were examined and their ability to predict subjective ratings of cough and cough-related quality of life were evaluated. In total, 70 subjects (mean+/-SD age 55+/-11.7 yrs, 51 (73%) females) with chronic unexplained cough (median duration 4.8 yrs, interquartile range 2.5-10.1 yrs) performed fully ambulatory 24-h sound recordings, which were manually counted by trained observers and quantified by 1) explosive phases, 2) cough seconds and 3) cough epochs. Subjects also completed cough visual analogue scales (VAS) and the Leicester Cough Questionnaire (LCQ). All units of cough were strongly correlated; explosive phases and cough seconds correlated slightly more strongly than cough seconds with cough epochs or explosive phases with cough epochs. LCQ scores correlated moderately with explosive phases and seconds; epochs correlated slightly less well. Cough VAS scores showed a similar pattern. Explosive phases and seconds are interchangeable units of cough, moderately related to subjective measures and cough-related quality of life; epochs are a less satisfactory alternative. PMID- 18287129 TI - Outcome of community-acquired pneumonia: influence of age, residence status and antimicrobial treatment. AB - Community-acquired pneumonia remains a major cause of mortality in developed countries. There is much discrepancy in the literature regarding factors influencing the outcome in the elderly population. Data were derived from a multicentre prospective study initiated by the German Competence Network for Community-Acquired Pneumonia. Patients with community-acquired pneumonia (n = 2,647; 1,298 aged < 65 yrs and 1,349 aged > or = 65 yrs) were evaluated, of whom 72.3% were hospitalised and 27.7% treated in the community. Clinical history, residence status, course of disease and antimicrobial treatment were prospectively documented. Microbiological investigations included cultures and PCR of respiratory samples and blood cultures. Factors related to mortality were included in multivariate analyses. The overall 30-day mortality was 6.3%. Elderly patients exhibited a significantly higher mortality rate that was independently associated with the following: age; residence status; confusion, urea, respiratory frequency and blood pressure (CURB) score; comorbid conditions; and failure of initial therapy. Increasing age remained predictive of death in the elderly. Nursing home residents showed a four-fold increased mortality rate and an increased rate of gram-negative bacillary infections compared with patients dwelling in the community. The CURB score and cerebrovascular disease were confirmed as independent predictors of death in this subgroup. Age and residence status are independent risk factors for mortality after controlling for comorbid conditions and disease severity. Failure of initial therapy was the only modifiable prognostic factor. PMID- 18287130 TI - Ciclesonide improves measures of small airway involvement in asthma. AB - Ciclesonide is delivered as a small-particle inhaled corticosteroid and improves lung function and airway hyperresponsiveness. The objective of the present study was to assess whether ciclesonide can specifically improve small airway function in asthma. A total of 16 mild-to-moderate asthma patients (seven males; median (range) age 39 (19-56) yrs and forced expiratory volume in one second (FEV(1)) 89 (62-120)% predicted) were randomised to 5 weeks' treatment with placebo or 320 mug ciclesonide once daily. The following small airway parameters were assessed: mean forced expiratory flow between 25 and 75% of forced vital capacity (FVC), percentage fall in FVC at provocative dose of adenosine-5'-monophosphate and of methacholine (MCh) causing a 20% fall in FEV(1), expiratory lung volume on computed tomography (CT) scan after MCh challenge, single-breath nitrogen closing volume and alveolar exhaled nitric oxide (eNO). Seven subjects received placebo and nine received ciclesonide. Both alveolar eNO and CT measurements of expiratory lung volume after MCh challenge decreased significantly with ciclesonide (median (range) decrease 4.4 (54.8-1.4) ppb and 59 (1,569- -117) mL, respectively), and compared with placebo (-0.4 (7.3- -3.4) ppb and -121 (20- 236) mL respectively). Ciclesonide did not significantly improve other small airways parameters. Inflammation and patency of small airways, reflected by alveolar exhaled nitric oxide and air trapping on computed tomography scan, both improve with ciclesonide even in this small number of patients. This indicates that ciclesonide exerts anti-inflammatory effects on small airways. PMID- 18287131 TI - Detection of somatic quantitative genetic alterations by multiplex polymerase chain reaction for the prediction of outcome in diffuse large B-cell lymphomas. AB - BACKGROUND: Genomic gains and losses play a crucial role in the development of diffuse large B-cell lymphomas. High resolution array comparative genomic hybridization provides a comprehensive view of these genomic imbalances but is not routinely applicable. We developed a polymerase chain reaction assay to provide information regarding gains or losses of relevant genes and prognosis in diffuse large B-cell lymphomas. DESIGN AND METHODS: Two polymerase chain reaction assays (multiplex polymerase chain reaction of short fluorescent fragments, QMPSF) were designed to detect gains or losses of c-REL, BCL6, SIM1, PTPRK, MYC, CDKN2A, MDM2, CDKN1B, TP53 and BCL2. Array comparative genomic hybridization was simultaneously performed to evaluate the sensitivity and predictive value of the QMPSF assay. The biological and clinical relevance of this assay were assessed. RESULTS: The predictive value of the QMPSF assay for detecting abnormal DNA copy numbers ranged between 88-97%, giving an overall concordance rate of 92% with comparative genomic hybridization results. In 77 cases of diffuse large B-cell lymphomas, gains of MYC, CDKN1B, c-REL and BCL2 were detected in 12%, 40%, 27% and 29%, respectively. TP53 and CDKN2A deletions were observed in 22% and 36% respectively. BCL2 and CDKN2A allelic status correlated with protein expression. TP53 mutations were associated with allelic deletions in 45% of cases. The prognostic value of a single QMPSF assay including TP53, MYC, CDKN2A, SIM1 and CDKN1B was predictive of the outcome independently of the germinal center B-cell like/non-germinal center B-cell like subtype or the International Prognostic Index. CONCLUSIONS: QMPSF is a reliable and flexible method for detecting somatic quantitative genetic alterations in diffuse large B-cell lymphomas and could be integrated in future prognostic predictive models. PMID- 18287132 TI - High response rate and improved graft-versus-host disease following bortezomib as salvage therapy after reduced intensity conditioning allogeneic stem cell transplantation for multiple myeloma. AB - We describe the results of 37 myeloma patients who received bortezomib following reduced intensity allogeneic stem cell transplantation (RIC-allo-SCT). Grade 1-2 peripheral neuropathy (35%), mild thrombocytopenia (24%) and fatigue (19%) were the most frequent adverse events, while there was no worsening of graft-vs-host disease symptoms. Twenty-seven patients (73%; 95% CI, 59-87%) achieved an objective response. With a median follow-up of 9 months from bortezomib initiation, the estimate of overall survival was 65% at 18 months while this was significantly higher (p=0.002) in the 27 patients achieving an objective response, suggesting that bortezomib is a safe and efficient option for myeloma patients after RIC-allo-SCT. PMID- 18287133 TI - Autoimmune thrombocytopenia in non-Hodgkin's lymphomas. AB - Autoimmune thrombocytopenia is a common immunehematologic complication in non Hodgkin's lymphomas and may complicate the treatment. We analyzed an original series from our institute as well as published cases of non-Hodgkin's lymphomas (excluding chronic lymphocytic leukemia) associated with autoimmune thrombocytopenia with regard to demographic factors, prevalence in non-Hodgkin's lymphoma subtypes and treatment outcome. The male/female ratio is 1.75. Half of the cases occurred prior to diagnosis of lymphoma. Chemotherapy is the best treatment in many non-Hodgkin's lymphomas patients with autoimmune thrombocytopenia compared with standard treatment of autoimmune thrombocytopenia. Splenectomy is effective in splenic marginal zone lymphoma. Autoimmune thrombocytopenia in patients with non-Hodgkin's lymphomas is potentially life threatening and difficult to treat. PMID- 18287134 TI - CD4+CD25+FOXP3+ T regulatory cells reconstitute and accumulate in the bone marrow of patients with multiple myeloma following allogeneic stem cell transplantation. AB - BACKGROUND: Very little is known about the number and function of immunosuppressive CD4(+)CD25(+)FOXP3(+) T regulatory cells (Treg) in the human bone marrow and it is unclear whether bone marrow-residing Treg are capable of regenerating following allogeneic stem cell transplantation. This is particularly surprising since the bone marrow represents a major priming site for T-cell responses and Treg play important roles in the prevention of T-cell-mediated graft-versus-host disease and in promoting tumor escape from T-cell-dependent immunosurveillance. DESIGN AND METHODS: Applying flow cytometry, real-time polymerase chain reaction, and functional assays, we performed the first study on bone marrow and peripheral blood Treg in healthy donors as well as multiple myeloma patients before and after allogeneic stem cell transplantation. RESULTS: We found that, following the allogeneic transplantation, donor-derived CD4(+)CD25(+)FOXP3(+) Treg expanded faster than conventional CD4(+) T cells, leading to an accumulation of Treg in the bone marrow of transplanted patients who lack relevant thymic function. The reconstituted bone marrow-residing CD4(+)CD25(+)FOXP3(+) Treg of myeloma patients after allogeneic stem cell transplantation consisted preferably of CD45RA(-)CCR7(-) memory T-cells and contained low numbers of T-cell receptor excision cycles, indicating that Treg had indeed expanded outside the thymus. Importantly, bone marrow-residing Treg of newly diagnosed and myeloma patients after allogeneic stem cell transplantation expressed high levels of transforming growth factor beta and cytotoxic T lymphocyte antigen 4, and showed a strong inhibitory function. CONCLUSIONS: We suggest that allogeneic stem cell transplantation provides a short but significant window of opportunity for CD8(+) T cells before an exuberant regeneration of immunosuppressive Treg sets in. Later after transplantation, bone marrow-residing Treg probably contribute to suppressing graft-versus-host disease but may also undermine persistent immune control of multiple myeloma. PMID- 18287135 TI - Normalization of the serum angiopoietin-1 to angiopoietin-2 ratio reflects response in refractory/resistant multiple myeloma patients treated with bortezomib. AB - Neoangiogenesis is involved in the pathophysiology of multiple myeloma and angiopoietins possibly contribute to myeloma-induced neovascularization. Bortezomib's antineoplastic potential includes an anti-angiogenic effect. We determined serum levels of angiopoietin-1 and angiopoietin-2 with ELISA pre- and post-bortezomib administration in 35 patients with relapsed/refractory multiple myeloma. Pre-bortezomib, serum angiopoietin-1 levels did not differ in patients and in healthy individuals, while serum angiopoietin-2 levels were elevated. Corresponding serum angiopoietin-1/angiopoietin-2 ratio was reduced in patients compared with controls. After treatment, serum angiopoietin-1 levels increased, while serum angiopoietin-2 levels decreased, therefore the angiopoietin 1/angiopoietin-2 ratio increased and normalized. This increase was significant in patients who responded to treatment. In conclusion, angiopoietin-1/angiopoietin-2 ratio normalization reflected response to bortezomib. PMID- 18287136 TI - Neurodegenerative central nervous system disease as late sequelae of Langerhans cell histiocytosis. Report from the Japan LCH Study Group. AB - Clinical features, brain magnetic resonance imaging findings and EDSS scores of 11 patients with neurodegenerative central nervous system Langerhans cell histiocytosis were analyzed in Japan. All patients initially had multi-system type Langerhans cell histiocytosis; 8 at 1-2 years of age and 3 at a later age. Neurodegenerative central nervous system Langerhans cell histiocytosis disease developed after a median time interval of 3.9 years from initial diagnosis. With a median follow-up of 4.5 years, 6 patients showed progression of disease with an EDSS score >3. This study demonstrates the importance of early detection of neurodegenerative central nervous system Langerhans cell histiocytosis by brain magnetic resonance imaging, particularly in the follow-up of patients who developed multi-system-type Langerhans cell histiocytosis in early infancy. PMID- 18287137 TI - Evaluation of the serum-free light chain test in untreated patients with AL amyloidosis. AB - We evaluated the Serum Free Light Chain (FLC) test in a series of 133 untreated patients with systemic AL amyloidosis. The FLC test detected the monoclonal gammopathy in 87% compared with 92% for immunofixation of serum and urine in combination. However, both tests proved complementary. The FLC test was also a valuable tool in patients with advanced renal failure in spite of uninvolved light chain retention. Higher FLC levels were associated with higher bone marrow plasmocytosis, poorer Karnofsky index and heart involvement, and therefore reflected disease severity. PMID- 18287138 TI - Clonal heterogeneity in chronic lymphocytic leukemia cells: superior response to surface IgM cross-linking in CD38, ZAP-70-positive cells. AB - BACKGROUND: Patients with chronic lymphocytic leukemia whose cells express CD38 and ZAP-70 and utilize unmutated Ig VH region genes have a very poor prognosis. We studied whether cells expressing CD38 and ZAP-70 are more susceptible to stimulation through B-cell receptors than are cells that do not express CD38 and ZAP-70. DESIGN AND METHODS: CD38-positive and CD38-negative leukemic cells were separated from single cases and compared for their response to B-cell receptor cross-linking and ZAP-70 expression. Cohort studies were also carried out by measuring the apoptotic response to surface immunoglobulin M (IgM) cross-linking in 82 patients with chronic lymphocytic leukemia and the protein tyrosine phosphorylation induced by surface IgM in 21 patients. RESULTS: CD38-positive cells, isolated from cases of chronic lymphocytic leukemia classified as CD38 positive or CD38-negative, expressed more ZAP-70 than the corresponding CD38 negative cells, exhibited more robust protein tyrosine phosphorylation and had a greater tendency to apoptosis upon B-cell receptor cross-linking. In the cohort studies, surface IgM-induced protein tyrosine phosphorylation correlated significantly with CD38 and ZAP-70 expression and with the absence of Ig VH gene mutations. Apoptosis induced by surface IgM cross-linking correlated significantly only with the proportion of CD38-positive cells. Difficulties in finding more definitive correlations were probably related to imprecision in the in vitro test system and in the definition of cases as positive or negative. CONCLUSIONS: Collectively, these data indicate that CD38-positive, ZAP-70 positive cells have a greater capacity for signaling through the B-cell receptor and suggest a function for B-cell receptor signaling in promoting chronic lymphocytic leukemia cell expansion, especially within the CD38-positive fraction of the leukemic clone. PMID- 18287139 TI - Vascular laboratory surveillance after intervention. Guest editorial. PMID- 18287140 TI - Duplex ultrasound surveillance can be worthwhile after arterial intervention. AB - A surveillance program based on duplex ultrasound testing after peripheral arterial intervention can increase long-term patency by identifying and by repairing clinical significant lesions. Its successful application requires numerous conditions regarding pathobiology of arterial repair failure and its consequences, arterial testing expertise, and durability of secondary procedures used to repair duplex-detected lesions. The methodology of surveillance should be tailored to the type of arterial intervention. Clinical reports on the efficacy of duplex ultrasound surveillance have supported its routine use, but controversy of cost-effectiveness remains. Duplex surveillance will decrease procedural primary patency, but when successful, the primary-assisted and secondary patency rates should be significantly higher than the rates when no surveillance was performed. An examination of the primary, primary-assisted, and secondary patency rates of an arterial procedure will indicate the benefit (or lack of benefit) of a surveillance program and the appropriateness of the threshold criteria used for secondary interventions. PMID- 18287142 TI - The application of duplex surveillance after carotid intervention. AB - Carotid duplex ultrasound testing provides a safe and accurate method to detect and grade the severity of atherosclerotic internal carotid artery stenosis both before and following carotid intervention. Testing after surgical endarterectomy or stent angioplasty allows assessment of the technical success by excluding residual stenosis. The focus of duplex surveillance after carotid intervention is to identify recurrent stenosis, repair site occlusion, and progression of contralateral internal carotid artery disease. Patients who develop a neurologic event or a duplex-detected >75% diameter-reducing internal carotid artery stenosis with a peak systolic velocity >300 cm/s and end-diastolic velocity >125 cm/s should be further evaluated by angiographic imaging and should be considered for reintervention if an appropriate lesion is confirmed. Duplex surveillance allows the vascular surgeon to evaluate patency of the rendered intervention, its stenosis-free durability, and its effectiveness in stroke prevention. PMID- 18287144 TI - Duplex surveillance of lower extremity endovascular interventions. AB - Endovascular techniques have been rapidly implemented into the surgical armamentarium for the treatment of infrainguinal arterial occlusive disease. Despite high initial technical success rates, all endovascular interventions in the femoropopliteal- tibial arterial distribution are associated with a significant restenosis and failure rate. Strategies to improve the durability of endovascular therapy need to be developed and should include medical therapy and surveillance testing to detect hemodynamic failure that typically precedes thrombotic occlusion. Duplex ultrasound can accurately identify the abnormal, stenosed angioplasty site, but the threshold criteria that predict thrombosis have not been defined. Duplex ultrasound surveillance protocols are designed to identify residual stenosis, which is associated with lower patency rates, and by serial testing, the development of a progressive, high-grade stenosis. The decision to reintervene and correct the angioplasty site lesion should be individualized based on patient characteristics, the clinical implication of angioplasty failure, and efficacy of secondary interventions in prolonging functional patency. PMID- 18287146 TI - Surveillance after lower extremity arterial bypass. AB - The elements of a surveillance program after lower limb bypass grafting to enhance graft patency continue to evolve. Graft evaluation should include clinical assessment for new or changes in limb ischemia symptoms, measurement of ankle or toe systolic pressure, or both, and duplex ultrasound imaging of the bypass graft, which in the early postoperative period is predictive of the subsequent need for bypass graft revision. The natural history of moderate graft stenosis is known, and these lesions can be safely monitored using serial testing to identify progression. The testing frequency should be individualized to the patient, type of arterial bypass, and duplex scan findings. Graft surveillance should focus on the identification and repair of critical stenosis (peak systolic velocity exceeding 300 cm/s, and peak systolic velocity ratio across the stenosis exceeding 3.5) correlating with more than 70% diameter-reducing stenosis. A graft surveillance program should result in a graft failure rate of less than 3% per year. PMID- 18287148 TI - Visceral duplex scanning: evaluation before and after artery intervention for chronic mesenteric ischemia. AB - Color duplex ultrasound testing has evolved to be a clinically useful modality to diagnose chronic mesenteric ischemia caused by visceral artery origin atherosclerosis. Testing requires expertise in ultrasound imaging, visceral artery hemodynamics, and duplex scan interpretation. Patient can be accurately screened for severe stenosis or occlusion involving celiac, superior mesenteric, or inferior mesenteric arteries. Duplex testing can also evaluate functional patency following visceral bypass grafting procedures or endovascular stent angioplasty. The focus of duplex surveillance after visceral artery intervention is to identify severe repair site stenosis, which can develop with symptoms of gut ischemia. Visceral duplex testing of a bypass graft or stent-angioplasty site that shows peak systolic velocities >300 cm/s with end-diastolic velocities >50 to 70 cm/s, or a decreased graft velocity peak systolic velocity <40 cm/s should be considered for interrogation using angiography to confirm or exclude severe (>70%) stenosis. Duplex testing after surgical or endovascular visceral interventions is a screening study, which compliments clinical follow-up by aiding the vascular surgeon in timely identification of visceral repairs that have developed a progressive, high-grade stenosis. PMID- 18287150 TI - Surveillance after endovascular abdominal aortic aneurysm repair. AB - (EVAR) Endovascular Aneurysm Repair of abdominal aortic aneurysms have mandated the need for surveillance imaging and other follow-up testing to minimize the complications of endograft failure and potential for abdominal aortic aneurysms rupture. The 2-dimensional and 3-dimensional contrast-enhanced computed tomography imaging currently serves as the gold standard for serial (EVAR) Endovascular Aneurysm Repair assessment, but this recommendation is being modified by successful clinical experience using duplex ultrasound, magnetic resonance imaging, and implantable wireless sac pressure sensors. Nearly all stent graft devices and abdominal aortic aneurysm-related complications after (EVAR) Endovascular Aneurysm Repair can be detected using available surveillance modalities assuming a compliant patient during follow-up. Alternating complementary surveillance techniques (contrast and noncontrast computed tomography, ultrasound, and pressure measures) can be used to reduce contrast dye and radiation exposure and can be tailored to patients with chronic renal insufficiency. PMID- 18287152 TI - Complications of carotid artery stenting are largely preventable: a retrospective error analysis. AB - Procedure-related complications with carotid artery stenting must be minimized for it to be a valid treatment for carotid stenosis. Failure analysis was done for 207 carotid stent procedures. All complications were reviewed and technical errors were identified. The procedure-related stroke rate was 2.9%, technical failure rate was 1.9%, and no patients died. Two strokes resulted from protocol deviations. A third stroke occurred while crossing a long, irregular lesion with a protection device instead of establishing flow reversal. Excessive instrumentation of the aortic arch resulted in 2 strokes. Distal embolization occurred after open-cell stenting a friable lesion in 1 patient. The majority of neurologic events and technical complications that occur during carotid stenting are preventable. By adhering to technical protocols, avoiding excessive instrumentation in the aortic arch, using flow reversal in selected lesions, and matching the appropriate anatomy and stent, results of carotid artery stenting can be exceptional. PMID- 18287153 TI - Commentary: has evidence changed practice? Appropriateness of carotid endarterectomy after the clinical trials. Halm EA, Tuhrim S, Wang JJ, Rojas M, Hannan EL, Chassin MR. Neurology. 2007;68:187-194. AB - available at http://www.neurology.org/cgi/ content/abstract/68/3/187. PMID- 18287154 TI - Commentary: risk factors for progression of peripheral arterial disease in large and small vessels. Aboyans V, Criqui MH, Denenberg JO, Knoke JD, Ridker PM, Fronek A. Circulation. 2006;113:2623-2629. AB - available at http://www.circ.ahajournals.org/cgi/ content/abstract/113/22/2623. PMID- 18287155 TI - COmmentary: assessment of renal artery stenosis severity by pressure gradient measurements. De Bruyne B, Manoharan G, Pijls NH, et al. J Am Coll Cardiol. 2006;48:1851-1855. AB - available at http://content.onlinejacc.org/cgi/ content/abstract/48/9/1851. PMID- 18287156 TI - Commentary: Nylaende M, Abdelnoor M, Stranden E, et al. The Oslo Balloon Angioplasty versus Conservative Treatment Study (OBACT)--the 2-years results of a single centre, prospective, randomised study in patients with intermittent claudication. Eur J Vasc Endovasc Surg. 2007;33:3-12. AB - available at http://www.medscape.com/medline/ abstract/17055756. PMID- 18287157 TI - Commentary: hyperglycemia independently increases the risk of perioperative stroke, myocardial infarction, and death after carotid endarterectomy. McGirt MJ, Woodworth GF, Brooke BS, et al. Neurosurgery. 2006;58:1066-1073. AB - available at http://www.medscape.com/medline/ abstract/16723885. PMID- 18287158 TI - Commentary: one-year cardiovascular event rates in outpatients with atherothrombosis. Steg PG, Bhatt DL, Wilson PW, et al; REACH Registry Investigators. JAMA. 2007;297: 1197-1206. AB - available at http://jama.ama-assn.org/cgi/ content/full/297/11/1197. PMID- 18287159 TI - Commentary: quality improvement in symptoms, functioning, and quality of life after peripheral endovascular revascularization. Safley DM, House JA, Laster SB, Daniel WC, Spertus JA, Marso SP. Circulation. 2007;115:550-552. AB - available at http://circ.ahajournals.org/cgi/ content/abstract/115/5/569. PMID- 18287161 TI - O-(beta-hydroxyethyl)-rutosides (Paroven; Venoruton): new studies, improved safety, and novel cardiovascular applications. PMID- 18287162 TI - O-(beta-hydroxyethyl)-rutosides systemic and local treatment in chronic venous disease and microangiopathy: an independent prospective comparative study. AB - O-(beta-hydroxyethyl)-rutosides (HR) is used to treat chronic venous disease and signs and symptoms of chronic venous insufficiency (CVI), varicose veins, and deep venous disease. This independent prospective controlled trial (a registry study) evaluates how the efficacy of HR at the local level (perimalleolar region) can be increased by the administration of a topical HR gel. The study is based on evaluation of microcirculatory variables in patients with severe CVI (ambulatory venous pressure, > 56 mm Hg) and venous microangiopathy. Patients are treated using 1 of the following 3 regimens: oral treatment with 1 g sachets of HR (2 g/d total) plus topical HR 2% gel applied 3 times daily at the internal perimalleolar region; oral treatment only (same dosage), or light elastic compression stockings. Laser Doppler skin flux at rest, skin flux at the perimalleolar region, and transcutaneous PO2 and PCO2 are measured at baseline and at the end of the treatment period. A comparable group of healthy individuals without treatment is observed for 8 weeks. In the treatment groups, flux is increased, PO2 is decreased, and PCO2 is increased compared with normal skin. At 4 and 8 weeks, the improvement in skin flux (which is decreased by all measurements), the increase in PO2, and the decrease in PCO2 (indicating microcirculatory improvement) are statistically significantly greater in the combined oral plus topical treatment group (P < .05). No adverse effects, tolerability problems, or compliance issues are noted. These results indicate an important role of HR in the treatment and control of CVI and venous microangiopathy. PMID- 18287163 TI - 5-Year control and treatment of edema and increased capillary filtration in venous hypertension and diabetic microangiopathy using O-(beta-hydroxyethyl) rutosides: a prospective comparative clinical registry. AB - This independent prospective controlled trial evaluates the efficacy of O-(beta hydroxyethyl)-rutosides (HR) during 5 years of administration against signs and symptoms and further degeneration of microcirculatory disturbances. The protective effect of HR in preventing end-point complications such as venous ulceration is evaluated. This study is based on evaluation of edema and the capillary filtration rate (CFR) in association with a clinical score scale. Patients having a severe degree of chronic venous insufficiency (CVI) and venous microangiopathy and completing at least 5 years of treatment are included. The following 4 groups are considered: group A (patients with CVI but without diabetes mellitus, receiving 1500 mg/d of HR), group B (patients with CVI and diabetes mellitus, receiving 2 g/d of HR), group C (control subjects receiving no pharmacologic or compression treatment), and group D (patients using elastic compression stockings only). All patients received the "best" available treatment. No adverse effects or intolerance is noted, with good compliance (>85%). In group A, there is a statistically significant decrease in the CFR during 5 years of follow-up. In group B, the decrease in the CFR is greater than that in group A. Reductions in edema, swelling, and the CFR during 5 years are notable, and values approach normal levels. During 5 years, HR is effective in treating venous edema and hypertension and in preventing deterioration of the distal venous system. The prevention of ulcerations with HR is another important observation. The effects of HR seem to be partially dose related, and tolerability and compliance are good. PMID- 18287164 TI - Efficacy of electroconvulsive therapy in the treatment of nondepressed psychiatric illness in elderly patients: a review of the literature. AB - Electroconvulsive therapy (ECT) is well established as a safe and effective treatment for several psychiatric disorders. Responsiveness to ECT does not abate with age, and data indicate that the use of ECT in the treatment of psychiatric disorders in the elderly persons has increased in recent decades. Special consideration must be given to the baseline cognitive abilities of an elderly patient prior to treatment with ECT. Much of the literature on the use of ECT in the elderly persons has focused on the treatment of mood disorders, whereas less research has been devoted to its use in the treatment of other psychiatric conditions. Although depressive syndromes remain the most common indication for ECT in the elderly persons, clinicians treating elderly patients should remain aware of the safety and efficacy of this treatment modality with other psychiatric disorders. This review examines the literature on the use of ECT in elderly patients with some common neuropsychiatric disorders including catatonia, bipolar mania, schizophrenia, dementia with behavioral disturbance, and Parkinson's disease. PMID- 18287165 TI - Lack of association between apolipoprotein E polymorphism and vascular dementia in Koreans. AB - To investigate an association of vascular dementia (VD) with the apolipoprotein E (APOE) polymorphism, the APOE polymorphism of 100 VD patients, 100 age- and gender-matched Alzheimer disease (AD) patients, and 200 age- and gender-matched nondemented control (NC) subjects was genotyped. The distribution of APOE polymorphism was compared. Neither the APOE epsilon4 allele nor the APOE epsilon2 allele was more prevalent in the VD patients compared with the NC subjects (P > .1 by the chi 2 test), which was the case when both men and women were analyzed separately (P > .1 by the chi2 test) and when young patients (75 years old or less) and old patients (more than 75 years old) were analyzed separately (P > .1 by the chi2 test). The estimated statistical power was over 0.80 when the odds ratios (OR) for VD conferred to the APOE epsilon4 are assumed to be higher than 2.2 and the type I error probability is set at 0.05, which is much higher than the power of the previous studies on the VD/APOE association. In conclusion, the results suggested that APOE epsilon4 allele does not confer the risk for VD, and even if it does, it does so very modestly. PMID- 18287166 TI - Computerized maze navigation and on-road performance by drivers with dementia. AB - This study examined the ability of computerized maze test performance to predict the road test performance of cognitively impaired and normal older drivers. The authors examined 133 older drivers, including 65 with probable Alzheimer disease, 23 with possible Alzheimer disease, and 45 control subjects without cognitive impairment. Subjects completed 5 computerized maze tasks employing a touch screen and pointer as well as a battery of standard neuropsychological tests. Parameters measured for mazes included errors, planning time, drawing time, and total time. Within 2 weeks, subjects were examined by a professional driving instructor on a standardized road test modeled after the Washington University Road Test. Road test total score was significantly correlated with total time across the 5 mazes. This maze score was significant for both Alzheimer disease subjects and control subjects. One maze in particular, requiring less than 2 minutes to complete, was highly correlated with driving performance. For the standard neuropsychological tests, highest correlations were seen with Trail Making A (TrailsA) and the Hopkins Verbal Learning Tests Trial 1 (HVLT1). Multiple regression models for road test score using stepwise subtraction of maze and neuropsychological test variables revealed significant independent contributions for total maze time, HVLT1, and TrailsA for the entire group; total maze time and HVLT1 for Alzheimer disease subjects; and TrailsA for normal subjects. As a visual analog of driving, a brief computerized test of maze navigation time compares well to standard neuropsychological tests of psychomotor speed, scanning, attention, and working memory as a predictor of driving performance by persons with early Alzheimer disease and normal elders. Measurement of maze task performance appears to be useful in the assessment of older drivers at risk for hazardous driving. PMID- 18287167 TI - Retention rates on RBANS memory subtests in elderly adults. AB - Identification of memory impairment is important for neuropsychological diagnostic and research applications, and retention rates on verbal and visual memory tests can provide useful information when characterizing a variety of neurological and psychiatric disorders. Although the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) is becoming a popular screening battery for cognitive functions, normative data on retention rates are not available. The retention rates of verbal and visual material were evaluated in a sample of clinical patients (n = 109) compared to a healthy control group (n = 718). Individual subtest retention rates were converted to age-corrected scaled scores based on the cumulative distribution of raw scores obtained by an elderly community-dwelling sample. Compared with the healthy normative sample, the percent retention found for the clinical group was significantly lower on all 3 RBANS memory subtests. These preliminary data suggest that retention rates of the RBANS memory subtests may add to the clinical utility of this test as a neuropsychological diagnostic and research tool. PMID- 18287168 TI - Neuropsychological correlates of self-reported depression and self-reported cognition among patients with mild cognitive impairment. AB - Patients with mild cognitive impairment (MCI) frequently experience significant depressive symptomatology and report cognitive disturbances. To date, no studies have examined the relationship between MCI patients' neuropsychologic functioning, self-reported depressive symptoms, and self-reported cognitive difficulties. In this study, 82 MCI patients completed a comprehensive neuropsychological evaluation that included the Beck Depression Inventory (BDI). A subset of 41 patients and informants also completed a measure of cognitive difficulties. Poorer memory functioning was associated with fewer self-reported depressive symptoms and fewer cognitive complaints. PMID- 18287169 TI - Stroke, depression, and functional health outcomes among adults in the community. AB - The objective was to determine the association between stroke and depression, the co-occurrence of stroke and depression, and functional health outcomes among adults in the US population. Data were drawn from the Midlife Development in the United States Study (MIDUS) (N = 3032), a nationally representative sample of adults age 25 to 74. Multiple logistic regression analyses were used to determine the association between stroke (past 12-month prevalence), depression (past 12 month prevalence), and functional health outcomes (past 12 months) and to determine whether there is an interaction between depression and stroke in predicting impairment in functioning. Almost one third (29.2%) of adults with stroke in the past year also had depression in the past year (odds ratio 3.5, 95% confidence interval 1.4, 8.9). The co-occurrence of stroke and depression was associated with significantly greater limitations in walking and climbing stairs and poorer general physical functioning than that associated with either without the other. There was evidence of interaction between depression and stroke in predicting limitations, specifically with a statistically significant effect in walking short distances (P = .045). These findings are consistent with previous clinical data and extend those data by suggesting that the co-occurrence of depression and stroke is associated with even poorer functional health outcomes, compared with each alone, and that there may be synergism between the two disorders related to this effect. PMID- 18287170 TI - Patterns of depression in Spanish- and English-speaking patients with Alzheimer's disease. AB - The purpose of this study was to evaluate whether distinct subtypes of depression could be identified in patients with Alzheimer's disease and, if so, to evaluate the patients in these subgroups. Ratings on the Cornell Scale for Depression in Dementia (CSDD) of 306 patients with Alzheimer's disease, 129 of whom were Spanish- and 177 English-speaking, were subjected to latent class analysis. Four subgroups were identified based on CSDD symptoms. These included an asymptomatic group, groups with mild and more severe typical depression, and a group characterized by prominent anxiety and irritability in addition to sadness. Group differences on demographic, cognitive, clinical, and functional status measures were explored via chi-square tests and analyses of variance. Results show that for some patients with Alzheimer's disease, patterns of symptoms of depression are similar to those in younger adult populations. A distinct subtype may exist, however, with prominent anxiety and irritability. PMID- 18287171 TI - Subclinical anxiety symptoms, sleep, and daytime dysfunction in older adults with primary insomnia. AB - "Both Insomnia complaints and anxiety-related distress are common in older adults, and are associated with poor daytime functioning. We investigated whether subclinical levels of anxiety were associated with sleep disturbance and daytime functioning in older adults who met diagnostic criteria for primary insomnia, and therefore but did not meet criteria for depression or an anxiety disorder. After adjustment for depressive symptoms, elevated state anxiety was associated with higher levels of wake after sleep onset (measured by both actigraphy and sleep log) and shorter sleep sleep onset latency (measured by sleep log). Higher levels of trait anxiety were associated with greater wake after sleep onset (measured by sleep log). Elevated state and trait anxiety were associated with worse and social functioning, and higher levels of trait anxiety were associated with worse role functioning. Thus, subclinical anxiety symptoms may be an important target for clinical intervention to improve sleep and functioning in older adults with primary insomnia." PMID- 18287172 TI - Posttraumatic stress and aggression among veterans in long-term care. AB - Because stresses associated with long-term care settings may exacerbate distress and aggression related to past trauma, we investigated self-report and staff reports of posttraumatic stress disorder symptoms and staff observations of verbal and physical aggression in 32 elderly males who were patients in a long term care unit for veterans. Feelings of anger and irritability were reported by 47% of patients; levels of anger and irritability were significantly correlated with observed aggressive behaviors (r = 0.43, P < .02); and observed aggressive behaviors were significantly more frequent among those reporting past traumatic stressors (t = 2.84, P < .008). Patient-reported posttraumatic stress disorder symptoms were significantly correlated with the frequency of past traumatic stressors (r = 0.48, P < .006). Observer-reported posttraumatic stress disorder symptoms and patient reports of anger were strongly correlated (r = 0.73, P < .001). No patient or staff reports were related to level of cognitive function. These findings are consistent with the hypothesis that posttraumatic symptoms can contribute to aggressive behaviors in elderly, medically ill, and cognitively impaired patients. PMID- 18287173 TI - Association between quality of life of demented patients and professional knowledge of care workers. AB - Professional knowledge about dementia and care methods is necessary for the appropriate care of demented people. In this study, the quality of life of 91 demented people staying at 12 care institutions was evaluated with the Quality of Life Questionnaire for Dementia (QOL-D). The amount of professional knowledge was evaluated in 140 staff members who took care of the patients using the professional knowledge test (PKT), and the mean PKT score was calculated for each institution (professional knowledge of institution). A positive significant correlation was observed between the QOL-D score and the professional knowledge of institution. The correlation remained significant when age, sex, and severity of dementia of demented patients were partialled out. This result indicated that the quality of life of demented people was better at institutions with staff members having more professional knowledge, indicating the importance of education of staff members and the possibility that such education improves the quality of life of the demented people. PMID- 18287174 TI - CytoPred: a server for prediction and classification of cytokines. AB - Cytokines are messengers of immune system. They are small secreted proteins that mediate and regulate the immune system, inflammation and hematopoiesis. Recent studies have revealed important roles played by the cytokines in adjuvants as therapeutic targets and in cancer therapy. In this paper, an attempt has been made to predict this important class of proteins and classify further them into families and subfamilies. A PSI-BLAST+Support Vector Machine-based hybrid approach is adopted to develop the prediction methods. CytoPred is capable of predicting cytokines with an accuracy of 98.29%. The overall accuracy of classification of cytokines into four families and further classification into seven subfamilies is 99.77 and 97.24%, respectively. It has been shown by comparison that CytoPred performs better than the already existing CTKPred. A user-friendly server CytoPred has been developed and available at http://www.imtech.res.in/raghava/cytopred. PMID- 18287175 TI - Thermostability promotes the cooperative function of split adenylate kinases. AB - Proteins can often be cleaved to create inactive polypeptides that associate into functional complexes through non-covalent interactions, but little is known about what influences the cooperative function of the ensuing protein fragments. Here, we examine whether protein thermostability affects protein fragment complementation by characterizing the function of split adenylate kinases from the mesophile Bacillus subtilis (AKBs) and the hyperthermophile Thermotoga neapolitana (AKTn). Complementation studies revealed that the split AKTn supported the growth of Escherichia coli with a temperature-sensitive AK, but not the fragmented AKBs. However, weak complementation occurred when the AKBs fragments were fused to polypeptides that strongly associate, and this was enhanced by a Q16L mutation that thermostabilizes the full-length protein. To examine how the split AK homologs differ in structure and function, their catalytic activity, zinc content, and circular dichroism spectra were characterized. The reconstituted AKTn had higher levels of zinc, greater secondary structure, and >10(3)-fold more activity than the AKBs pair, albeit 17 fold less active than full-length AKTn. These findings provide evidence that the design of protein fragments that cooperatively function can be improved by choosing proteins with the greatest thermostability for bisection, and they suggest that this arises because hyperthermophilic protein fragments exhibit greater residual structure compared to their mesophilic counterparts. PMID- 18287176 TI - An improved prediction of catalytic residues in enzyme structures. AB - The protein databases contain a huge number of function unknown proteins, including many proteins with newly determined 3D structures resulted from the Structural Genomics Projects. To accelerate experiment-based assignment of function, de novo prediction of protein functional sites, like active sites in enzymes, becomes increasingly important. Here, we attempted to improve the prediction of catalytic residues in enzyme structures by seeking and refining different encodings (i.e. residue properties) as well as employing new machine learning algorithms. In particular, considering that catalytic residues can often reveal specific network centrality when representing enzyme structure as a residue contact network, the corresponding measurement (i.e. closeness centrality) was used as one of the most important encodings in our new predictor. Meanwhile, a genetic algorithm integrated neural network (GANN) was also employed. Thanks to the above strategies, our GANN predictor demonstrated a high accuracy of 91.2% in the prediction of catalytic residues based on balanced datasets (i.e. the 1:1 ratio of catalytic to non-catalytic residues). When the GANN method was optimally applied to real enzyme structures, 73.9% of the tested structures had the active site correctly located. Compared with two existing methods, the proposed GANN method also demonstrated a better performance. PMID- 18287177 TI - A novel random mutagenesis approach using human mutagenic DNA polymerases to generate enzyme variant libraries. AB - The in vitro MutaGen procedure is a new random mutagenesis method based on the use of low-fidelity DNA polymerases. In the present study, this technique was applied on a 2 kb gene encoding amylosucrase, an attractive enzyme for the industrial synthesis of amylose-like polymers. Mutations were first introduced during a single replicating step performed by mutagenic polymerases pol beta and pol eta. Three large libraries (>10(5) independent clones) were generated (one with pol beta and two with pol eta). The sequence analysis of randomly chosen clones confirmed the potential of this strategy for the generation of diversity. Variants generated by pol beta were 4-7-fold less mutated than those created with pol eta, indicating that our approach enables mutation rate control following the DNA polymerase employed for mutagenesis. Moreover, pol beta and pol eta provide different and complementary mutation spectra, allowing a wider sequence space exploration than error-prone PCR protocols employing Taq polymerase. Interestingly, some of the variants generated by pol eta displayed unusual modifications, including combinations of base substitutions and codon deletions which are rarely generated using other methods. By taking advantage of the mutation bias of naturally highly error-prone DNA polymerases, MutaGen thus appears as a very useful tool for gene and protein randomisation. PMID- 18287178 TI - Aerobic reserve and physical functional performance in older adults. AB - BACKGROUND: older adults can be limited in their performance of daily tasks due to an inadequate aerobic capacity. Aerobic capacity below minimum physiological thresholds required to maintain independence leaves older adults with little, or no, aerobic reserve. OBJECTIVE: the aim of this study was to measure functional performance and aerobic reserve in older adults during the serial performance of daily tasks. SUBJECTS: twenty-nine (n = 29) men and women (n = 23 females) 70-92 years of age participated in this study. METHODS: performance based physical function was assessed using the Continuous-Scale Physical Functional Performance test (CS-PFP). A Cosmed K4b(2) portable metabolic system was used to measure VO(2PEAK) and oxygen uptake during the serial performance of a battery of daily tasks (VO(2PFP)). Aerobic reserve was calculated as the difference between VO(2PEAK) and VO(2PFP). RESULTS: the correlation coefficient between aerobic reserve and functional performance was r = 0.50(P = 0.006). Participants utilized 32.2 +/- 8.1%, 42.7 +/- 10.8%, and 50.3 +/- 12.3% of VO(2PEAK) for the low, moderate, and high workloads of the CS-PFP, respectively. CONCLUSIONS: light housework and carrying groceries require 40 to 50% of peak oxygen consumption. This information can be used by clinicians and health professionals working with older adults as a guide to how much aerobic fitness is needed to perform ADLs and maintain independence. These can then be used as guides for assessment and for setting training goals in older adults. PMID- 18287179 TI - Association between global leukocyte DNA methylation, renal function, carotid intima-media thickness and plasma homocysteine in patients with stage 2-4 chronic kidney disease. AB - BACKGROUND: Patients with chronic kidney disease (CKD) have an increased risk of cardiovascular disease (CVD). Preliminary evidence suggests a role for global DNA hypomethylation in the pathogenesis of atherosclerotic complications in CKD. The aims of this study in patients with stage 2-4 CKD were (1) to assess the association between renal function and DNA methylation, (2) to assess the association between DNA methylation and two markers of atherosclerosis [common carotid intima-media thickness (CCA-IMT)] and brachial artery endothelium dependent, flow-mediated dilatation (BA-FMD) and (3) to examine the effect of a multi-step treatment strategy on DNA methylation. METHODS: In the Anti-Oxidant Therapy In Chronic Renal Insufficiency study (ATIC-study), 93 patients with stage 2-4 CKD were included. In a randomized, double-blind, placebo-controlled design, the treatment group received pravastatin to which vitamin E was added after 6 months and homocysteine-lowering B-vitamin therapy after another 6 months. DNA methylation was assessed using tandem mass spectrometry. CCA-IMT and BA-FMD were assessed using B-mode ultrasonography. RESULTS: At baseline, global DNA methylation was not associated with the estimated glomerular filtration rate (P = 0.32) or with CCA-IMT (P = 0.62) or BA-FMD (P = 0.51). No effect of the treatment strategy including B-vitamin on global DNA methylation was found either in the total study group or within separate strata of homocysteine concentration and renal function. CONCLUSION: In patients with stage 2-4 CKD, global DNA methylation is not associated with renal function or with CCA-IMT or BA-FMD. A treatment strategy that includes B-vitamins did not alter global DNA methylation in these patients. These data do not support the role of DNA hypomethylation in CKD-associated vascular disease in patients with stage 2-4 CKD. PMID- 18287180 TI - What we know (and do not know) about raising children with complex continuing care needs. PMID- 18287181 TI - Understanding the views of parents of children with special needs about the nursing care their child receives when in hospital: a qualitative study. AB - This article reports a qualitative study designed to explore parents' views on how their child with additional needs had been cared for by hospital nursing staff, focusing on how well their own and their child's needs had been identified and met. Twelve interviews with parents of children with additional needs and a thematic analysis of the interview data was conducted. Four themes were developed to provide an insight into parents' views about their experiences: their prior experiences of hospital care, including the process of being told the original diagnosis; communication with staff; nurse-parent relationships; and perceptions of nurses and nursing care. It concludes that parents experience some difficulties in developing a trusting relationship with the nurses caring for their child with additional needs. Parents perceive that nurses are not always able to recognize and respond to their needs when caring for their child. Failure to address these needs can interfere with the development of effective nurse parent relationships. PMID- 18287182 TI - Using participant observation in pediatric health care settings: ethical challenges and solutions. AB - Participant observation strategies may be particularly effective for research involving children and their families in health care settings. These techniques, commonly used in ethnography and grounded theory, can elicit data and foster insights more readily than other research approaches, such as structured interviews or quantitative methods. This article outlines recommendations for the ethical conduct of participant observation in pediatric health care settings. This involves a brief overview of the significant contributions that participant observation can bring to our understanding of children and families in health care settings; an examination of the elements of participant observation that are necessary conditions for its effective conduct; an outline of contemporary ethical norms in Canada, the United Kingdom and the United States for research in pediatric health care settings; and a discussion of how participant observation research should be operationalized in order to comply with these norms. PMID- 18287183 TI - Children and young people's preference of thematic design and colour for their hospital environment. AB - In this innovative project, the views of children and young people were explored regarding their preference of thematic design and colour for their hospital environment in a new children's unit. The novelty of the approach was that it was driven by the preferred choices of children and young people through the use of 'child-friendly' interviews and questionnaires. Informing the study was the development of a group of children and young people who underwent research training, and with support, developed all data collection tools and helped to verify data analysis. A two-phased sequential study was undertaken. During phase 1, 40 interviews were performed with children and young people, including 10 with additional learning needs and physical disabilities while 140 questionnaires were analysed for phase 2 of the study. Notable issues emerged about preferred thematic designs of walls, doors and floors, while new findings were revealed regarding colour preferences for wards, entrances and outpatient areas. PMID- 18287184 TI - Gender assignment surgery on children with disorders of sex development: a case report and discussion from South Africa. AB - This case report highlights the dilemma faced by staff with regard to the timing of surgery on a child with a disorder of sex development living in a large, lower socio-economic class, South African, urban township. In this community, children with disorders of sex development can sometimes become an object of interest and ridicule or are thought to be bewitched. Many parents of children with such disorders find it difficult to protect their offspring from the marginalization and rejection that is the consequence of such curiosity and transparency. Current research and theory pertaining to the biological and social bases of gender identity and behaviour are reviewed and their capacity to guide decisions is explored. The absence of a support group to assist these children and their parents, and the paucity of information available in the public domain, compounds an already challenging problem. PMID- 18287185 TI - The needs of parents of hospitalized children in Australia. AB - The aim of this study was to compare the perceptions of needs held by parents of hospitalized children with those held by the staff caring for them. Family centred care is a central tenet of paediatrics and should encompass all aspects of the child and family's experience of hospitalization. Important to this are the needs of parents when their children are hospitalized. A widely used and validated tool was used with a convenience sample in paediatric facilities in a children's hospital in Australia. Some differences were found between parents and staff for scores for perceived importance of the 51 needs included in the questionnaire, and whether or not they were being met satisfactorily during children's hospital admission, although there were no consistent patterns. Parents declared themselves more independent than the staff perceived them to be. These findings facilitate improvements in communication between parents and staff and can be included in education programmes for both. PMID- 18287186 TI - Prospective study of peer victimization and social-psychological adjustment in children with endocrine disorders. AB - This article evaluates the relations between peer victimization and child and parent reports of social-psychological variables 1.5 years later. Thirty-six children diagnosed with endocrine disorders and their parents completed questionnaires regarding social-psychological functioning. Peer victimization at time 2 was significantly related to concurrent depression, loneliness, externalizing and internalizing symptoms. Hierarchical linear regression equations indicated that peer victimization at baseline was not a significant predictor of time 2 social-psychological functioning when baseline levels of each variable were controlled. Significant correlations were found between baseline and time 2 levels of social anxiety, loneliness, externalizing and internalizing symptoms, with medium to large effect sizes. Peer victimization, social anxiety, loneliness, internalizing and externalizing behavior problems are relatively stable across time. Peer victimization is related to concurrent adjustment problems but may not contribute to the development of new problems. Early identification and intervention is imperative. Medical visits are an opportunity to assess and refer for treatment. PMID- 18287187 TI - Quantification of risk-benefit in interventional radiology. AB - The increased use of interventional radiological procedures brings with it an increased risk of cancer induction due to the possible high radiation levels used. This risk must be balanced against any viable alternatives and should take into account the individual risks and benefits. The purpose of this study is to quantify this risk benefit in terms of quality-adjusted life years (QALYs) of a procedure and its alternatives, including the risk due to the radiation. Uterine fibroid embolisation was chosen as a model study with data collected both on the outcomes of procedures in terms of complications and quality of life in the long term and on radiation dose delivered to the patient. Dosimetry data were obtained from University Hospital Galway. The risk was then calculated using the BEIR VII model of dose-response for low linear energy transfer (LET) radiation. In this case, it was found that the radiological technique has distinct advantages over hysterectomy and myomectomy in terms of QALYs. The increased risk due to radiation from the procedure is small when compared with the overall benefit to the patient. Probability of stochastic effects based on the average dose per procedure carries an attributable risk (fatal and non-fatal cancer and hereditary effects) of 0.4% or a relative risk of 1.09%. The method described here can readily be adapted to other interventional procedures. PMID- 18287188 TI - Staff dosimetry in interventional cardiology: survey on methods and level of exposure. AB - In interventional cardiac procedures, staff operates near the patient in a non uniformly scattered radiation field. Consequently, workers may receive, over a period, relatively high radiation doses. The measurement of individual doses to personnel becomes critical due to the use of protective devices and, as a consequence of the large number of methods proposed to assess the effective dose, great variability in monitoring programmes is expected among European countries. SENTINEL consortium has conducted a survey on staff dosimetry methods and on the level of staff exposure in 12 European cardiac centres demonstrating the urgent need to harmonise dosimetry methods. From the dosimetry survey, constraint annual effective dose of 1.4 mSv and Hp(0.07) over the protective apron of 14 mSv are proposed for the optimisation the exposure the most-exposed operator. PMID- 18287189 TI - Patient dose in interventional radiology: a European survey. AB - Patient doses for a few common fluoroscopy-guided procedures in interventional radiology (IR) (excluding cardiology) were collected from a few radiological departments in 13 European countries. The major aim was to evaluate patient doses for the basis of the reference levels. In total, data for 20 procedures for about 1300 patients were collected. There were many-fold variations in the number of IR equipment and procedures per population, in the entrance dose rates, and in the patient dose data (total dose area product or DAP, fluoroscopy time and number of frames). There was no clear correlation between the total DAP and entrance dose rate, or between the total DAP and fluoroscopy time, indicating that a number of parameters affect the differences. Because of the limited number of patients, preliminary reference levels were proposed only for a few procedures. There is a need to improve the optimisation of IR procedures and their definitions and grouping, in order to account for their different complexities. PMID- 18287190 TI - Proposal for a patient database on cardiac interventional exposures for epidemiological studies. AB - Relatively high organ doses absorbed by patients in interventional cardiology suggest the opportunity to define these patients as a cohort to be followed forward in time in an epidemiological study of the cancer risks associated with exposure to low-dose ionising radiation. In this paper, the UNSCEAR 2000 Report risk estimates for the most exposed organs/tissues in cardiac interventional procedures are reviewed, as well as the critical features of such an epidemiological study that is anticipated to have an intrinsically low statistical power because of the low levels of risk and possible confounding factors. To overcome these limitations, data collected in different institutions can be combined provided that a common design and conduct are used for dose assessment. A minimum dataset to be collected on a patient basis is proposed that can be implemented routinely in most facilities. This data should be linked to the local patient information system in order to retrieve all the exposures of a given patient. PMID- 18287191 TI - Effects of radiologists' skill and experience on patient doses in interventional examinations. AB - In this study, effects of radiologists' skill and experience on patient doses were investigated. Dose-area product and entrance surface doses of two groups of patients, one examined by a number of junior radiologists and another one by a senior radiologist, have been compared for the diagnostic interventional examinations of cerebral and lower limbs. Collimation of the X-ray beam and shortening the fluoroscopy times by the senior radiologist considerably reduced the patient doses for interventional cerebral examinations. PMID- 18287192 TI - Patient dosimetry in paediatric diagnostic radiology. AB - A study of patient exposure in paediatric diagnostic radiology was conducted in three dedicated paediatric departments. The entrance surface dose was assessed by three methods: direct measurement by thermoluminescent dosemeters, calculation from the dose-area product and from the tube output. The results obtained by the these methods were compared, proving that all of them are applicable for the assessment of paediatric doses. Subsequently, the accumulated data were compared with the mean values from other similar studies and to the reference dose levels set by the Commission of European Communities, which clearly demonstrated the need for further investigation of the paediatric dose levels in Bulgaria and for optimisation of the radiological practice. PMID- 18287193 TI - Improving evaluation of primary gastric malignancies by distending the stomach with milk immediately before 18F-FDG PET scanning. AB - This study was designed to investigate the feasibility and effectiveness of a modified protocol for (18)F-FDG PET that was proposed to improve the identification of primary gastric malignancies. METHODS: In the modified protocol, patients were asked to drink 300-500 mL of cow milk to distend the stomach immediately before PET scans instead of fasting all along. For investigation of the influence of ingested milk on (18)F-FDG distributions, 43 nondiabetic patients without documented gastric diseases underwent both empty- and distended-stomach PET scans (79 and 72 scans, respectively) in their serial follow-up studies. For the evaluation of proven gastric malignancies, 24 patients who underwent distended-stomach PET scans were compared with 17 patients who underwent conventional empty-stomach examinations. RESULTS: Ingestion of milk nearly 1 h after (18)F-FDG injection had no significant influence on distributions to the heart (P = 0.16), mediastinum (P = 0.50), and liver (P = 0.49), whereas the percentages of intense and moderate uptake in the stomach changed from 38.0% and 59.5% to 0% and 11.1%, respectively. With the normal gastric wall distended, malignant lesions were observed with higher contrast and clearer outlines, and some of them were detected at a small size (1.2 cm) at an early stage and with mild uptake. CONCLUSION: Gastric distention with milk just before (18)F-FDG PET is a simple and effective method for improving the evaluation of primary gastric malignancies. PMID- 18287194 TI - Optimized energy window configuration for 201Tl imaging. AB - A poor signal-to-noise ratio attributable to a low injected dose of thallium and the presence of scattered photons are the major impediments in the use of thallium as an imaging agent. Thallium decays in a complicated way and emits photons in a wide range of energies (68-82 keV). To increase the ratios of primary photons to scatter photons (primary-to-scatter ratios) and possibly increase system sensitivity, a new energy window for thallium was investigated. METHODS: The NCAT phantom was used to simulate the distribution of activity and the attenuation coefficient in a typical patient torso. The phantom was imaged with a SPECT simulator in different energy window configurations. The energy spectra for primary photons and scatter photons were generated, and the most suitable energy windows were investigated. To evaluate the results of the simulation study, a physical phantom was imaged in different energy windows with a SPECT system. The images of the physical phantom were analyzed for the best quality image and the corresponding window setting. To evaluate the windows determined in the simulation and phantom studies, SPECT images of 7 patients who had angiographically confirmed myocardial defects were acquired in different energy windows. The images were quantitatively compared on the basis of the calculated contrast, scatter-to-noise ratio, and sensitivity. The images were also qualitatively evaluated independently by 4 nuclear medicine specialists. RESULTS: The simulation study showed that the conventional window setting (68 +/- 10% keV) is not the most suitable window configuration for (201)Tl imaging and that the optimum energy window is 77 +/- 15% keV. The images acquired in the latter window configuration yielded higher primary-to-scatter ratios, higher sensitivity (total counts), and better contrast than the images acquired in the conventional window configuration. The phantom study confirmed the results of the simulation study. In the clinical study, the images acquired in the suggested window showed a considerable increase in myocardium-to-defect contrast (1.541 +/- 0.368) and myocardium-to-cavity contrast (1.171 +/- 0.099) than those acquired in the conventional window configuration. CONCLUSION: The energy window configuration of 77 +/- 15% keV yields higher-quality images than the conventional window configuration. PMID- 18287195 TI - Review of common occupational hazards and safety concerns for nuclear medicine technologists. AB - The purpose of this article is to address common occupational hazards and safety concerns of nuclear medicine technologists. There are many possible occupational hazards, but this review is intended to concentrate on common hazards and safety concerns. These include radiation safety issues and concerns about the possibility of developing latent diseases, such as eye cataracts or cancer; pregnant workers and radiation safety issues; biohazard concerns associated with patient body fluids; possible low-back pain from moving heavy equipment and performing patient transfers; and possible repetitive trauma disorders, such as carpal tunnel syndrome, from computer work. Suggestions are made regarding how to identify potential hazards and avoid them. After reading this article, nuclear medicine technologists should be able to explain the importance of the as-low-as reasonably-achievable concept, discuss the possible effects of ionizing radiation on the adult and the developing fetus, list several basic principles to avoid injury to the back, list and describe the more common repetitive trauma disorders or injuries and how to avoid them, and list and describe the biohazard safety issues that nuclear medicine technologists face and how to develop policy to minimize exposure risk. PMID- 18287196 TI - SPECT/CT physical principles and attenuation correction. AB - Using nuclear medicine techniques, physiologic activity and processes can be identified in a way that is unique from other modalities. Oftentimes it is helpful to know the exact location of the physiologic uptake that is visualized on a scan. Knowing the exact location can sometimes help to distinguish normal from abnormal physiologic uptake. When an abnormality has been identified, knowing the exact location can then be helpful in treatment planning. The ability to provide precise localization of physiologic data from nuclear medicine studies is now possible with hybrid SPECT/CT systems. Additionally, these systems provide an accurate attenuation correction of the nuclear medicine image data. After reading this article, the technologist will be able to list and describe the inherent problems associated with SPECT image acquisition and reconstruction, briefly explain how data acquired from the CT scanner are used to provide attenuation correction data for SPECT and anatomic information for diagnostic purposes, list and briefly describe the different types of clinical SPECT/CT systems, and discuss the importance of accurate CT and SPECT image registration. PMID- 18287197 TI - Consensus recommendations for gastric emptying scintigraphy: a joint report of the American Neurogastroenterology and Motility Society and the Society of Nuclear Medicine. AB - This consensus statement from the members of the American Neurogastroenterology and Motility Society and the Society of Nuclear Medicine recommends a standardized method for measuring gastric emptying (GE) by scintigraphy. A low fat, egg-white meal with imaging at 0, 1, 2, and 4 h after meal ingestion, as described by a published multicenter protocol, provides standardized information about normal and delayed GE. Adoption of this standardized protocol will resolve the lack of uniformity of testing, add reliability and credibility to the results, and improve the clinical utility of the GE test. PMID- 18287198 TI - Cardiac CT: indications and limitations. AB - Cardiac CT (CCT) is rapidly evolving as a noninvasive imaging modality. Newer technologic developments in CCT allow the comprehensive assessment of cardiovascular anatomy, including the coronary arteries. There are special considerations regarding obtaining and accurately interpreting CCT studies. On completion of this article, the reader should be able to describe the issues related to adequate patient preparation and acquisition and interpretation of CCT studies, recognize specific limitations that impair image quality and subsequently the accuracy of diagnosis, and list the current indications and future potential applications of this technique. PMID- 18287199 TI - Silencing of Smed-betacatenin1 generates radial-like hypercephalized planarians. AB - Little is known about the molecular mechanisms responsible for axis establishment during non-embryonic processes such as regeneration and homeostasis. To address this issue, we set out to analyze the role of the canonical Wnt pathway in planarians, flatworms renowned for their extraordinary morphological plasticity. Canonical Wnt signalling is an evolutionarily conserved mechanism to confer polarity during embryonic development, specifying the anteroposterior (AP) axis in most bilaterians and the dorsoventral (DV) axis in early vertebrate embryos. beta-Catenin is a key element in this pathway, although it is a bifunctional protein that is also involved in cell-cell adhesion. Here, we report the characterization of two beta-catenin homologs from Schmidtea mediterranea (Smed betacatenin1/2). Loss of function of Smed-betacatenin1, but not Smed betacatenin2, in both regenerating and intact planarians, generates radial-like hypercephalized planarians in which the AP axis disappears but the DV axis remains unaffected, representing a unique example of a striking body symmetry transformation. The radial-like hypercephalized phenotype demonstrates the requirement for Smed-betacatenin1 in AP axis re-establishment and maintenance, and supports a conserved role for canonical Wnt signalling in AP axis specification, whereas the role of beta-catenin in DV axis establishment would be a vertebrate innovation. When considered alongside the protein domains present in each S. mediterranea beta-catenin and the results of functional assays in Xenopus embryos demonstrating nuclear accumulation and axis induction with Smed betacatenin1, but not Smed-betacatenin2, these data suggest that S. mediterranea beta-catenins could be functionally specialized and that only Smed-betacatenin1 is involved in Wnt signalling. PMID- 18287200 TI - Mesenchyme-dependent BMP signaling directs the timing of mandibular osteogenesis. AB - To identify molecular and cellular mechanisms that determine when bone forms, and to elucidate the role played by osteogenic mesenchyme, we employed an avian chimeric system that draws upon the divergent embryonic maturation rates of quail and duck. Pre-migratory neural crest mesenchyme destined to form bone in the mandible was transplanted from quail to duck. In resulting chimeras, quail donor mesenchyme established significantly faster molecular and histological programs for osteogenesis within the relatively slower-progressing duck host environment. To understand this phenotype, we assayed for changes in the timing of epithelial mesenchymal interactions required for bone formation and found that such interactions were accelerated in chimeras. In situ hybridization analyses uncovered donor-dependent changes in the spatiotemporal expression of genes, including the osteo-inductive growth factor Bmp4. Mesenchymal expression of Bmp4 correlated with an ability of quail donor cells to form bone precociously without duck host epithelium, and also relied upon epithelial interactions until mesenchyme could form bone independently. Treating control mandibles with exogenous BMP4 recapitulated the capacity of chimeras to express molecular mediators of osteogenesis prematurely and led to the early differentiation of bone. Inhibiting BMP signaling delayed bone formation in a stage-dependent manner that was accelerated in chimeras. Thus, mandibular mesenchyme dictates when bone forms by temporally regulating its interactions with epithelium and its own expression of Bmp4. Our findings offer a developmental mechanism to explain how neural crest-derived mesenchyme and BMP signaling underlie the evolution of species-specific skeletal morphology. PMID- 18287201 TI - An Arabidopsis F-box protein acts as a transcriptional co-factor to regulate floral development. AB - Plants flower in response to both environmental and endogenous signals. The Arabidopsis LEAFY (LFY) transcription factor is crucial in integrating these signals, and acts in part by activating the expression of multiple floral homeotic genes. LFY-dependent activation of the homeotic APETALA3 (AP3) gene requires the activity of UNUSUAL FLORAL ORGANS (UFO), an F-box component of an SCF ubiquitin ligase, yet how this regulation is effected has remained unclear. Here, we show that UFO physically interacts with LFY both in vitro and in vivo, and this interaction is necessary to recruit UFO to the AP3 promoter. Furthermore, a transcriptional repressor domain fused to UFO reduces endogenous LFY activity in plants, supporting the idea that UFO acts as part of a transcriptional complex at the AP3 promoter. Moreover, chemical or genetic disruption of proteasome activity compromises LFY-dependent AP3 activation, indicating that protein degradation is required to promote LFY activity. These results define an unexpected role for an F-box protein in functioning as a DNA associated transcriptional co-factor in regulating floral homeotic gene expression. These results suggest a novel mechanism for promoting flower development via protein degradation and concomitant activation of the LFY transcription factor. This mechanism may be widely conserved, as homologs of UFO and LFY have been identified in a wide array of plant species. PMID- 18287202 TI - Ascl1 is required for oligodendrocyte development in the spinal cord. AB - Development of oligodendrocytes, myelin-forming glia in the central nervous system (CNS), proceeds on a protracted schedule. Specification of oligodendrocyte progenitors (OLPs) begins early in development, whereas their terminal differentiation occurs at late embryonic and postnatal periods. How these distinct steps are controlled remains unclear. Our previous study demonstrated an important role of the helix-loop-helix (HLH) transcription factor Ascl1 in early generation of OLPs in the developing spinal cord. Here, we show that Ascl1 is also involved in terminal differentiation of oligodendrocytes late in development. Ascl1-/- mutant mice showed a deficiency in differentiation of myelin-expressing oligodendrocytes at birth. In vitro culture studies demonstrate that the induction and maintenance of co-expression of Olig2 and Nkx2-2 in OLPs, and thyroid hormone-responsive induction of myelin proteins are impaired in Ascl1 /- mutants. Gain-of-function studies further showed that Ascl1 collaborates with Olig2 and Nkx2-2 in promoting differentiation of OLPs into oligodendrocytes in vitro. Overexpression of Ascl1, Olig2 and Nkx2-2 alone stimulated the specification of OLPs, but the combinatorial action of Ascl1 and Olig2 or Nkx2-2 was required for further promoting their differentiation into oligodendrocytes. Thus, Ascl1 regulates multiple aspects of oligodendrocyte development in the spinal cord. PMID- 18287204 TI - CDC-25.1 stability is regulated by distinct domains to restrict cell division during embryogenesis in C. elegans. AB - Cdc25 phosphatases are key positive cell cycle regulators that coordinate cell divisions with growth and morphogenesis in many organisms. Intriguingly in C. elegans, two cdc-25.1(gf) mutations induce tissue-specific and temporally restricted hyperplasia in the embryonic intestinal lineage, despite stabilization of the mutant CDC-25.1 protein in every blastomere. We investigated the molecular basis underlying the CDC-25.1(gf) stabilization and its associated tissue specific phenotype. We found that both mutations affect a canonical beta-TrCP phosphodegron motif, while the F-box protein LIN-23, the beta-TrCP orthologue, is required for the timely degradation of CDC-25.1. Accordingly, depletion of lin-23 in wild-type embryos stabilizes CDC-25.1 and triggers intestinal hyperplasia, which is, at least in part, cdc-25.1 dependent. lin-23(RNAi) causes embryonic lethality owing to cell fate transformations that convert blastomeres to an intestinal fate, sensitizing them to increased levels of CDC-25.1. Our characterization of a novel destabilizing cdc-25.1(lf) intragenic suppressor that acts independently of lin-23 indicates that additional cues impinge on different motifs of the CDC-25.1 phosphatase during early embryogenesis to control its stability and turnover, in order to ensure the timely divisions of intestinal cells and coordinate them with the formation of the developing gut. PMID- 18287203 TI - Lrig3 regulates neural crest formation in Xenopus by modulating Fgf and Wnt signaling pathways. AB - Leucine-rich repeats and immunoglobulin-like domains 3 (Lrig3) was identified by microarray analysis among genes that show differential expression during gastrulation in Xenopus laevis. Lrig3 was expressed in the neural plate and neural crest (NC) at neurula stages, and in NC derivatives and other dorsal structures during tailbud stages. A prominent consequence of the morpholino induced inhibition of Lrig3 expression was impaired NC formation, as revealed by the suppression of marker genes, including Slug, Sox9 and Foxd3. In the NC induction assay involving Chordin plus Wnt3a-injected animal caps, Lrig3 morpholino inhibited expression of Slug, Sox9 and Foxd3, but not of Pax3 and Zic1. In line with this, Lrig3 knockdown prevented NC marker induction by Pax3 and Zic1, suggesting that Lrig3 acts downstream of these two genes in NC formation. Injection of Lrig3 and Wnt3a led to low-level induction of NC markers and enhanced induction of Fgf3, Fgf4 and Fgf8 in animal caps, suggesting a positive role for Lrig3 in Wnt signaling. Lrig3 could attenuate Fgf signaling in animal caps, did interact with Fgf receptor 1 in cultured cells and, according to context, decreased or increased the induction of NC markers by Fgf. We suggest that Lrig3 functions in NC formation in Xenopus by modulating the Wnt and Fgf signaling pathways. PMID- 18287205 TI - Mutation of DNA primase causes extensive apoptosis of retinal neurons through the activation of DNA damage checkpoint and tumor suppressor p53. AB - Apoptosis is often observed in developing tissues. However, it remains unclear how the apoptotic pathway is regulated during development. To clarify this issue, we isolated zebrafish mutants that show extensive apoptosis of retinal cells during their development. pinball eye (piy) is one such mutant, in which retinal stem cells proliferate normally but almost all retinal neurons undergo apoptosis during differentiation. We found that a missense mutation occurred in the small subunit of DNA primase (Prim1) in the piy mutant. DNA primase is essential for DNA replication; however, this mutation does not affect cell proliferation but rather induces neuronal apoptosis. RNA synthesis catalyzed by Prim1 is important for the activation of the DNA damage response, which may activate Ataxia telangiectasia mutated (ATM), Checkpoint kinase 2 (Chk2) and the tumor suppressor p53. We found that the apoptosis induced by the prim1 mutation depends on the ATM Chk2-p53 apoptotic pathway. These data suggest that the surveillance system of genome integrity strongly influences the cell fate decision between differentiation and apoptosis during retinal neurogenesis in zebrafish. PMID- 18287206 TI - The growing catalog of small RNAs and their association with distinct Argonaute/Piwi family members. AB - Several distinct classes of small RNAs, some newly identified, have been discovered to play important regulatory roles in diverse cellular processes. These classes include siRNAs, miRNAs, rasiRNAs and piRNAs. Each class binds to distinct members of the Argonaute/Piwi protein family to form ribonucleoprotein complexes that recognize partially, or nearly perfect, complementary nucleic acid targets, and that mediate a variety of regulatory processes, including transcriptional and post-transcriptional gene silencing. Based on the known relationship of Argonaute/Piwi proteins with distinct classes of small RNAs, we can now predict how many new classes of small RNAs or silencing processes remain to be discovered. PMID- 18287207 TI - Substrate-dependent effects of human ABCB1 coding polymorphisms. AB - One of the many obstacles to effective drug treatment is the efflux transporter P glycoprotein (P-gp), which can restrict the plasma and intracellular concentrations of numerous xenobiotics. Variable drug response to P-gp substrates suggests that genetic differences in ABCB1 may affect P-gp transport. The current study examined how ABCB1 variants alter the P-gp-mediated transport of probe substrates in vitro. Nonsynonymous ABCB1 variants and haplotypes with an allele frequency >/=2% were transiently expressed in HEK293T cells, and the transport of calcein acetoxymethyl ester and 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY-FL)-paclitaxel was measured in the absence or presence of the P-gp inhibitor cyclosporin A. The A893S, A893T, and V1251I variants and the N21D/1236C>T/A893S/3435C>T haplotype altered intracellular accumulation compared with reference P-gp in a substrate-dependent manner. It is interesting that certain variants showed altered sensitivity to cyclosporin A inhibition that was also substrate-specific. These functional data demonstrate that nonsynonymous polymorphisms in ABCB1 may selectively alter P-gp transport and drug-drug interactions in a substrate- and inhibitor-dependent manner. PMID- 18287208 TI - Cytokeratin-RNA cross-linking mediated by the antitumor aminoflavone, 5-amino-2,3 fluorophenyl-6,8-difluoro-7-methyl-4H-1-benzopyran-4-one. AB - Aminoflavone (AF) is an anticancer drug in early clinical trials, and its antiproliferative activity involves the induction of DNA-protein cross-links. To identify the proteins cross-linked to nucleic acids, cesium chloride (CsCl) gradient centrifugation was used to isolate proteins tightly bound to nucleic acids in AF-treated human breast carcinoma MCF-7 cells. The identified proteins included structural proteins (several cytokeratins), transcription regulators, and stress response proteins. The identification of the cytokeratins was validated using direct immunoblotting of the high-density CsCl (nucleic acid) fractions isolated from AF-treated cells. Ribonuclease A pretreatment caused the cytokeratin signal in the heaviest CsCl fractions to disappear, suggesting that AF mediates RNA-cytokeratin cross-links. Additional experiments using radiolabeled AF showed that AF formed adducts with total RNA and mRNA with similar affinity to that of DNA. Moreover, 18S RNA was selectively pulled down using an anti-cytokeratin antibody after AF treatment. Consistent with the formation of these adducts, we found that AF inhibits RNA and protein synthesis in a dose- and time-dependent manner. This study provides evidence for the formation of AF-mediated cytokeratin-RNA cross-links and the presence of cytokeratin-RNA complexes. Thus, in addition to its anticancer activity, AF might be a useful molecular probe to study the potential role of cytokeratins in the subcellular localization and metabolism of RNA. PMID- 18287209 TI - Cardioprotective and survival benefits of long-term combined therapy with beta2 adrenoreceptor (AR) agonist and beta1 AR blocker in dilated cardiomyopathy postmyocardial infarction. AB - We have reported therapeutic effectiveness of pharmacological stimulation of beta2 adrenoreceptors (ARs) to attenuate the cardiac remodeling and myocardial infarction (MI) expansion in a rat model of dilated cardiomyopathy (DCM) post-MI. Furthermore, the combination of beta2 AR stimulation with beta1 AR blockade exceeded the therapeutic effectiveness of beta1 AR blockade. However, these studies were relatively short (6 weeks). In this study, in the same experimental model, we compared different effects, including survival benefit, of combined therapy with the beta1 AR blocker, metoprolol, plus the beta2 AR agonist, fenoterol (beta1-beta2+), and either therapy alone (beta1- or beta2+) during the 1-year study. Therapy was started 2 weeks after permanent ligation of the left coronary artery. Cardiac remodeling, MI expansion, and left ventricular function were assessed by serial echocardiography and compared with untreated animals (nT). Sixty-seven percent mortality in nT was reduced to 33% in the beta1-beta2+ (p < 0.01). Progressive cardiac remodeling observed in nT and beta1- was significantly attenuated in beta1-beta2+ during the first 6 months of treatment. In beta1-beta2+, MI expansion was completely prevented, and functional decline was significantly attenuated during the entire year. Myocardial apoptosis was significantly reduced in both beta1-beta2+ and beta1-. A reduction of cardiac beta1 AR density and decreases in chronotropic and contractile responses to beta2 AR-specific stimulation in the absence of a reduction of beta2 AR density in nT were precluded in rats receiving combined therapy. The results demonstrate the cardioprotective and survival benefit of long-term combination therapy of beta2 AR agonists and beta1 AR blockers in a model of DCM. PMID- 18287210 TI - MF498 [N-{[4-(5,9-Diethoxy-6-oxo-6,8-dihydro-7H-pyrrolo[3,4-g]quinolin-7-yl)-3 methylbenzyl]sulfonyl}-2-(2-methoxyphenyl)acetamide], a selective E prostanoid receptor 4 antagonist, relieves joint inflammation and pain in rodent models of rheumatoid and osteoarthritis. AB - Previous evidence has implicated E prostanoid receptor 4 (EP4) in mechanical hyperalgesia induced by subplantar inflammation. However, its role in chronic arthritis remains to be further defined because previous attempts have generated two conflicting lines of evidence, with one showing a marked reduction of arthritis induced by a collagen antibody in mice lacking EP4, but not EP1-EP3, and the other showing no impact of EP4 antagonism on arthritis induced by collagen. Here, we assessed the effect of a novel and selective EP4 antagonist MF498 [N-{[4-(5,9-diethoxy-6-oxo-6,8-dihydro-7H-pyrrolo[3,4-g]quinolin-7-yl)-3 methylbenzyl]sulfonyl}-2-(2-methoxyphenyl)acetamide] on inflammation in adjuvant induced arthritis (AIA), a rat model for rheumatoid arthritis (RA), and joint pain in a guinea pig model of iodoacetate-induced osteoarthritis (OA). In the AIA model, MF498, but not the antagonist for EP1, MF266-1 [1-(5-{3-[2-(benzyloxy)-5 chlorophenyl]-2-thienyl}pyridin-3-yl)-2,2,2-trifluoroethane-1,1-diol] or EP3 MF266-3 [(2E)-N-[(5-bromo-2-methoxyphenyl)sulfonyl]-3-[5-chloro-2-(2 naphthylmethyl)phenyl]acrylamide], inhibited inflammation, with a similar efficacy as a selective cyclooxygenase 2 (COX-2) inhibitor MF-tricyclic. In addition, MF498 was as effective as an nonsteroidal anti-inflammatory drug, diclofenac, or a selective microsomal prostaglandin E synthase-1 inhibitor, MF63 [2-(6-chloro-1H-phenanthro[9,10-d]imidazol-2-yl)isophthalonitrile], in relieving OA-like pain in guinea pigs. When tested in rat models of gastrointestinal toxicity, the EP4 antagonist was well tolerated, causing no mucosal leakage or erosions. Lastly, we evaluated the renal effect of MF498 in a furosemide-induced diuresis model and demonstrated that the compound displayed a similar renal effect as MF-tricyclic [3-(3,4-difluorophenyl)-4-(4-(methylsulfonyl)phenyl)-2 (5H)-furanone], reducing furosemide-induced natriuresis by approximately 50%. These results not only suggest that EP4 is the major EP receptor in both RA and OA but also provide a proof of principle to the concept that antagonism of EP4 may be useful for treatment of arthritis. PMID- 18287211 TI - Modeling the binding and function of metabotropic glutamate receptors. AB - A mathematical model for the binding and function of metabotropic glutamate receptors was developed, with the aim to gain new insights into the functioning of these complex receptors. These receptors are homodimers, and each subunit is composed of a ligand binding [Venus flytrap (VFT)] domain and a heptahelical domain (HD) responsible for G-protein activation. Our mechanistic model integrates all structural information available so far: the various states of the VFT dimer (open-open, closed-open, and closed-closed), as well as the fact that a single HD is active at a time. To provide the model with parameters with biological meaning, two published experimental studies were reanalyzed. The first one reports a negative cooperativity in agonist binding (J Biol Chem 279:35526 35534, 2004), whereas the other indicates a positive cooperativity in agonist mediated response (Nat Struct Mol Biol 11:706-713, 2004). The former study allowed us to explain the mechanistic features associated with VFT recognition by agonists and antagonists integrating a negative allosteric interaction for agonist binding. The second study helped us to quantitatively describe the functional dynamics of transduction of the VFT occupation into functional response, confirming a putative positive cooperativity at the level of receptor coupling efficacy. This model will help both to better understand the functioning of these receptors and to characterize the mechanism of action of various types of allosteric modulators. Moreover, this model may be of general utility for oligomeric systems in which the ligand binding and effector domains correspond to distinct structural domains. PMID- 18287212 TI - In vivo activation of peroxisome proliferator-activated receptor-delta protects the heart from ischemia/reperfusion injury in Zucker fatty rats. AB - Peroxisome proliferator-activated receptor (PPAR)-delta is a transcription factor that belongs to the PPAR family. PPAR-delta is abundantly expressed in the heart, and its role in the heart is largely unknown. We tested whether pharmacological activation of PPAR-delta protects the heart from ischemia/reperfusion (I/R) injury in male Zucker fatty rats, a rodent model of obesity and dyslipidemia. A highly selective PPAR-delta agonist, [4-[[[2-[3-fluoro-4-(trifluoromethyl)phenyl] 4-methyl-5-thiazolyl]methyl] thio]-2-methylphenoxy]acetic acid (GW0742), was administered for 7 days at 10 mg/kg/day (p.o., once a day). Ischemic injury was produced by occlusion of the left anterior descending artery for 30 min followed by reperfusion for up to 24 h. Treatment with GW0742 reduced serum levels of cardiac troponin-I and infarct size by 63% (p < 0.01) and 32% (p < 0.01), respectively, and improved left ventricular function. Treatment with GW0742 up regulated gene expression involved in cardiac fatty acid oxidation, increased fat use in the heart, and reduced serum levels of free fatty acids. The enhanced cardiac expression of interleukin (IL)-6, IL-8, intercellular adhesion molecule 1, and monocyte chemoattractant protein-1 induced by I/R were significantly attenuated by GW0742. Treatment with GW0742 also reduced apoptotic cardiomyocytes by 34% and cardiac caspase-3 activity by 61% (both p < 0.01 versus vehicle). GW0742 differentially regulated Bcl family members, favoring cell survival, and attenuated I/R-induced cardiac mitochondrial damage. In addition, GW0742 treatment augmented the cardiac Akt signaling pathway, as reflected by enhanced phospho-3-phosphoinositide-dependent kinase-1 and p-Akt. The results indicate that activation of PPAR-delta protected the heart from I/R injury in Zucker fatty rats, and multiple mechanisms including amelioration of lipotoxicity, anti inflammation, and up-regulation of prosurvival signaling contribute together to the cardioprotection. PMID- 18287213 TI - Delivery of acid sphingomyelinase in normal and niemann-pick disease mice using intercellular adhesion molecule-1-targeted polymer nanocarriers. AB - Type B Niemann-Pick disease (NPD) is a multiorgan system disorder caused by a genetic deficiency of acid sphingomyelinase (ASM), for which lung is an important and challenging therapeutic target. In this study, we designed and evaluated new delivery vehicles for enzyme replacement therapy of type B NPD, consisting of polystyrene and poly(lactic-coglycolic) acid polymer nanocarriers targeted to intercellular adhesion molecule (ICAM)-1, an endothelial surface protein up regulated in many pathologies, including type B NPD. Real-time vascular imaging using intravital microscopy and postmortem imaging of mouse organs showed rapid, uniform, and efficient binding of fluorescently labeled ICAM-1-targeted ASM nanocarriers (anti-ICAM/ASM nanocarriers) to endothelium after i.v. injection in mice. Fluorescence microscopy of lung alveoli actin, tissue histology, and 125I albumin blood-to-lung transport showed that anti-ICAM nanocarriers cause neither detectable lung injury, nor abnormal vascular permeability in animals. Radioisotope tracing showed rapid disappearance from the circulation and enhanced accumulation of anti-ICAM/125I-ASM nanocarriers over the nontargeted naked enzyme in kidney, heart, liver, spleen, and primarily lung, both in wild-type and ASM knockout mice. These data demonstrate that ICAM-1-targeted nanocarriers may enhance enzyme replacement therapy for type B NPD and perhaps other lysosomal storage disorders. PMID- 18287214 TI - Preclinical characterization of selective phosphodiesterase 10A inhibitors: a new therapeutic approach to the treatment of schizophrenia. AB - We have recently proposed the hypothesis that inhibition of the cyclic nucleotide phosphodiesterase (PDE) 10A may represent a new pharmacological approach to the treatment of schizophrenia (Curr Opin Invest Drug 8:54-59, 2007). PDE10A is highly expressed in the medium spiny neurons of the mammalian striatum (Brain Res 985:113-126, 2003; J Histochem Cytochem 54:1205-1213, 2006; Neuroscience 139:597 607, 2006), where the enzyme is hypothesized to regulate both cAMP and cGMP signaling cascades to impact early signal processing in the corticostriatothalamic circuit (Neuropharmacology 51:374-385, 2006; Neuropharmacology 51:386-396, 2006). Our current understanding of the physiological role of PDE10A and the therapeutic utility of PDE10A inhibitors derives in part from studies with papaverine, the only pharmacological tool for this target extensively profiled to date. However, this agent has significant limitations in this regard, namely, relatively poor potency and selectivity and a very short exposure half-life after systemic administration. In the present report, we describe the discovery of a new class of PDE10A inhibitors exemplified by TP-10 (2-{4-[-pyridin-4-yl-1-(2,2,2-trifluoro-ethyl)-1H-pyrazol-3-yl] phenoxymethyl}-quinoline succinic acid), an agent with greatly improved potency, selectivity, and pharmaceutical properties. These new pharmacological tools enabled studies that provide further evidence that inhibition of PDE10A represents an important new target for the treatment of schizophrenia and related disorders of basal ganglia function. PMID- 18287216 TI - Chronic intermittent hypoxia reduces ventilatory long-term facilitation and enhances apnea frequency in newborn rats. AB - Ventilatory long-term facilitation (LTF; defined as gradual increase of minute ventilation following repeated hypoxic exposures) is well described in adult mammals and is hypothesized to be a protective mechanism against apnea. In newborns, LTF is absent during the first postnatal days, but its precise developmental pattern is unknown. Accordingly, this study describes this pattern of postnatal development. Additionally, we tested the hypothesis that chronic intermittent hypoxia (CIH) from birth alters this development. LTF was estimated in vivo using whole body plethysmography by exposing rat pups at postnatal days 1, 4, and 10 (P1, P4, and P10) to 10 brief hypoxic cycles (nadir 5% O2) and respiratory recordings during the following 2 h (recovery, 21% O2). Under these conditions, ventilatory LTF (gradual increase of minute ventilation during recovery) was clearly expressed in P10 rats but not in P1 and P4. In a second series of experiments, rat pups were exposed to CIH during the first 10 postnatal days (6 brief cyclic exposures at 5% O2 every 6 min followed by 1 h under normoxia, 24 h a day). Compared with P10 control rats, CIH enhanced hypoxic ventilatory response (estimated during the hypoxic cycles) specifically in male rat pups. Ventilatory LTF was drastically reduced in P10 rats exposed to CIH, which was associated with higher apnea frequency during recovery. We conclude that CIH from birth enhances hypoxic chemoreflex and disrupts LTF development, thus likely contributing to increase apnea frequency. PMID- 18287217 TI - Sex and the renin-angiotensin system: inequality between the sexes in response to RAS stimulation and inhibition. AB - The purpose of this review is to examine sex differences in response to stimulation and inhibition of the renin-angiotensin system (RAS). The RAS plays a prominent role in the development of chronic renal disease, and there are known sex differences not only in the expression level of components of the RAS but also in how males and females respond to perturbations of the RAS. In men, renal injury increases in parallel with increased activation of the RAS, while in women, increases in ANG II do not necessarily translate into increases in renal injury. Moreover, both epidemiological and experimental studies have noted sex differences in the therapeutic benefits following angiotensin-converting enzyme inhibitor and angiotensin receptor blocker treatment. Despite these differences, RAS inhibitors are the most commonly prescribed drugs for the treatment of chronic renal disease, irrespective of sex. This review will examine how males and females respond to stimulation and inhibition of the RAS, with a focus on renal disease. PMID- 18287215 TI - Effect of chronic endothelin receptor antagonism on cerebrovascular function in type 2 diabetes. AB - Diabetes increases the risk of stroke and contributes to poor clinical outcomes in this patient population. Myogenic tone of the cerebral vasculature, including basilar arteries, plays a key role in controlling cerebral blood flow. Increased myogenic tone is ameliorated with ET receptor antagonism in Type 1 diabetes. However, the role of endothelin-1 (ET-1) and its receptors in cerebrovascular dysfunction in Type 2 diabetes, a common comorbidity in stroke patients, remains poorly elucidated. Therefore, we hypothesized that 1) cerebrovascular dysfunction occurs in the Goto-Kakizaki (GK) model of Type 2 diabetes, and 2) pharmacological antagonism of ETA receptors ameliorates, while ETB receptor blockade augments vascular dysfunction. GK or control rats were treated with antagonists to either ETA (atrasentan, 5 mg.kg(-1).day(-1)) or ETB (A-192621, 15 or 30 mg.kg(-1).day( 1)) receptors for 4 wk and vascular function of basilar arteries was assessed using a wire myograph. GK rats exhibited increased sensitivity to ET-1. ET(A) receptor antagonism caused a rightward shift, indicating decreased sensitivity in diabetes, while it increased sensitivity to ET-1 in control rats. Endothelium dependent relaxation was impaired in diabetes. ETA receptor blockade restored relaxation to control values in the GK animals with no significant effect in Wistar rats and ETB blockade with 30 mg.kg(-1).day(-1) A-192621 caused paradoxical constriction in diabetes. These studies demonstrate that cerebrovascular dysfunction occurs and may contribute to altered regulation of myogenic tone and cerebral blood flow in diabetes. While ETA receptors mediate vascular dysfunction, ETB receptors display differential effects. These results underscore the importance of ETA/ETB receptor balance and interactions in cerebrovascular dysfunction in diabetes. PMID- 18287218 TI - Sleep deprivation can inhibit adult hippocampal neurogenesis independent of adrenal stress hormones. AB - Sleep deprivation (SD) can suppress cell proliferation in the hippocampal dentate gyrus of adult male rodents, suggesting that sleep may contribute to hippocampal functions by promoting neurogenesis. However, suppression of cell proliferation in rats by the platform-over-water SD method has been attributed to elevated corticosterone (Cort), a potent inhibitor of cell proliferation and nonspecific correlate of this procedure. We report here results that do not support this conclusion. Intact and adrenalectomized (ADX) male rats were subjected to a 96-h SD using multiple- and single-platform methods. New cells were identified by immunoreactivity for 5-bromo-2'-deoxyuridine (BrdU) or Ki67 and new neurons by immunoreactivity for BrdU and doublecortin. EEG recordings confirmed a 95% deprivation of rapid eye movement (REM) sleep and a 40% decrease of non-REM sleep. Cell proliferation in the dentate gyrus was suppressed by up to 50% in sleep-deprived rats relative to apparatus control or home cage control rats. This effect was also observed in ADX rats receiving continuous low-dose Cort replacement via subcutaneous minipumps but not in ADX rats receiving Cort replacement via drinking water. In these latter rats, Cort intake via water was reduced by 60% during SD; upregulation of cell proliferation by reduced Cort intake may obscure inhibitory effects of sleep loss on cell proliferation. SD had no effect on the percentage of new cells expressing a neuronal phenotype. These results demonstrate that the Cort replacement method is critical for detecting an effect of SD on cell proliferation and support a significant role for sleep in adult neurogenesis. PMID- 18287219 TI - Osmoregulatory fluid intake but not hypovolemic thirst is intact in mice lacking angiotensin. AB - Water intakes in response to hypertonic, hypovolemic, and dehydrational stimuli were investigated in mice lacking angiotensin II as a result of deletion of the angiotensinogen gene (Agt-/- mice), and in C57BL6 wild-type (WT) mice. Baseline daily water intake in Agt-/- mice was approximately threefold that of WT mice because of a renal developmental disorder of the urinary concentrating mechanisms in Agt-/- mice. Intraperitoneal injection of hypertonic saline (0.4 and 0.8 mol/l NaCl) caused a similar dose-dependent increase in water intake in both Agt-/- and WT mice during the hour following injection. As well, Agt-/- mice drank appropriate volumes of water following water deprivation for 7 h. However, Agt-/- mice did not increase water or 0.3 mol/l NaCl intake in the 8 h following administration of a hypovolemic stimulus (30% polyethylene glycol sc), whereas WT mice increased intakes of both solutions during this time. Osmoregulatory regions of the brain [hypothalamic paraventricular and supraoptic nuclei, median preoptic nucleus, organum vasculosum of the lamina terminalis (OVLT), and subfornical organ] showed an increased number of neurons exhibiting Fos-immunoreactivity in response to intraperitoneal hypertonic NaCl in both Agt-/- mice and WT mice. Polyethylene glycol treatment increased Fos-immunoreactivity in the subfornical organ, OVLT, and supraoptic nuclei in WT mice but only increased Fos immunoreactivity in the supraoptic nucleus in Agt-/- mice. These data show that brain angiotensin is not essential for the adequate functioning of neural pathways mediating osmoregulatory thirst. However, angiotensin II of either peripheral or central origin is probably necessary for thirst and salt appetite that results from hypovolemia. PMID- 18287220 TI - Experimental autoimmune prostatitis induces chronic pelvic pain. AB - Pain is the hallmark of patients with chronic prostatitis (CP) and chronic pelvic pain syndrome (CPPS). Despite numerous hypotheses, the etiology and pathogenesis remain unknown. To better understand CP/CPPS, we used a murine experimental autoimmune prostatitis model to examine the development, localization, and modulation of pelvic pain. Pelvic pain was detected 5 days after antigen instillation and was sustained beyond 30 days, indicating the development of chronic pain. The pain was attenuated by lidocaine treatment into the prostate, but not into the bladder or the colon, suggesting that pain originated from the prostate. Experimental autoimmune prostatitis histopathology was confined to the prostate with focal periglandular inflammatory infiltrates in the ventral, dorsolateral, and anterior lobes of the mouse prostate. Inflammation and pelvic pain were positively correlated and increased with time. Morphologically, the dorsolateral prostate alone showed significantly increased neuronal fiber distribution, as evidenced by increased protein gene product 9.5 expression. Pelvic pain was attenuated by treatment with the neuromodulator gabapentin, suggesting spinal and/or supraspinal contribution to chronic pain. These results provide the basis for identifying mechanisms that regulate pelvic pain and the testing of therapeutic agents that block pain development in CP/CPPS. PMID- 18287221 TI - Weight regain after sustained weight reduction is accompanied by suppressed oxidation of dietary fat and adipocyte hyperplasia. AB - A dual-tracer approach (dietary 14C-palmitate and intraperitoneal 3H-H2O) was used to assess the trafficking of dietary fat and net retention of carbon in triglyceride depots during the first 24 h of weight regain. Obesity-prone male Wistar rats were allowed to mature under obesogenic conditions for 16 wk. One group was switched to ad libitum feeding of a low-fat diet for 10 wk (Obese group). The remaining rats were switched to an energy-restricted, low-fat diet for 10 wk that reduced body weight by 14% and were then assessed in energy balance (Reduced group), with free access to the low-fat diet (Relapse-Day1 group), or with a provision that induced a minor imbalance (+10 kcal) equivalent to that observed in obese rats (Gap-Matched group). Fat oxidation remained at a high, steady rate throughout the day in Obese rats, but was suppressed in Reduced, Gap-Matched, and Relapse-Day1 rats though 9, 18, and 24 h, respectively. The same caloric excess in Obese and Gap-Matched rats led to less fat oxidation over the day and greater trafficking of dietary fat to visceral depots in the latter. In addition to trafficking nutrients to storage, Relapse-Day1 rats had more small, presumably new, adipocytes at the end of 24 h. Dietary fat oxidation at 24 h was related to the phosphorylation of skeletal muscle acetyl-CoA carboxylase and fatty acid availability. These observations provide evidence of adaptations in the oxidation and trafficking of dietary fat that extend beyond the energy imbalance, which facilitate rapid, efficient regain during the relapse to obesity. PMID- 18287223 TI - Simultaneous pulmonary trunk and pulmonary arterial wave intensity analysis in fetal lambs: evidence for cyclical, midsystolic pulmonary vasoconstriction. AB - The physiological basis of a characteristically low blood flow to the fetal lungs is incompletely understood. To determine the potential role of pulmonary vascular interaction in this phenomenon, simultaneous wave intensity analysis (WIA) was performed in the pulmonary trunk (PT) and left pulmonary artery (LPA) of 10 anesthetized late-gestation fetal sheep instrumented with PT and LPA micromanometer catheters to measure pressure (P) and transit-time flow probes to obtain blood velocity (U). Studies were performed at rest and during brief complete occlusion of the ductus arteriosus to augment pulmonary vasoconstriction (n = 4) or main pulmonary artery to abolish wave transmission from the lungs (n = 3). Wave intensity (dI(W)) was calculated as the product of the P and U rates of change. Forward and backward components of dI(W) were determined after calculation of wave speed. PT and LPA WIA displayed an early systolic forward compression wave (FCW(is)) increasing P and U, and a late systolic forward expansion wave decreasing P and U. However, a marked midsystolic fall in LPA U to near-zero was related to an extremely prominent midsystolic backward compression wave (BCW(ms)) that arose approximately 5 cm distal to the LPA, was threefold larger than the PT BCW(ms) (P < 0.001), of similar size to FCW(is) at rest (P > 0.6), larger than FCW(is) following ductal occlusion (P < 0.05) and abolished after main pulmonary artery occlusion. These findings suggest that the absence of pulmonary arterial midsystolic forward flow which accompanies a low fetal lung blood flow is due to a BCW(ms) generated in part by cyclical vasoconstriction within the pulmonary microcirculation. PMID- 18287222 TI - Endogenous sodium pump inhibitors and age-associated increases in salt sensitivity of blood pressure in normotensives. AB - Factors that mediate increases in salt sensitivity of blood pressure with age remain to be clarified. The present study investigated 1) the effects of high NaCl intake on two Na pump inhibitors, endogenous ouabain (EO) and marinobufagenin (MBG), in middle-aged and older normotensive Caucasian women; and 2) whether individual differences in EO and MBG are linked to variations in sodium excretion or salt sensitivity. A change from 6 days of a lower (0.7 mmol.kg(-1).day(-1))- to 6 days of a higher (4 mmol.kg(-1).day(-1))-NaCl diet elicited a sustained increase in MBG excretion that directly correlated with an increase in the fractional Na excretion and was inversely related to age and to an age-dependent increase in salt sensitivity. In contrast, EO excretion increased only transiently in response to NaCl loading and did not vary with age or correlate with fractional Na excretion or salt sensitivity. A positive correlation of both plasma and urine levels of EO and MBG during salt loading may indicate a casual link between two Na pump inhibitors in response to NaCl loading, as observed in animal models. A linear mixed-effects model demonstrated that age, dietary NaCl, renal MBG excretion, and body mass index were each independently associated with systolic blood pressure. Thus, a sustained increase in MBG in response to acutely elevated dietary NaCl is inversely linked to salt sensitivity in normotensive middle-aged and older women, and a relative failure of MBG elaboration by these older persons may be involved in the increased salt sensitivity with advancing age. PMID- 18287224 TI - Medullary pathways mediating the parasubthalamic nucleus depressor response. AB - The parasubthalamic nucleus (PSTN) projects extensively to the nucleus of the solitary tract (NTS); however, the function of PSTN in cardiovascular regulation is unknown. Experiments were done in alpha-chloralose anesthetized, paralyzed, and artificially ventilated rats to investigate the effect of glutamate (10 nl, 0.25 M) activation of PSTN neurons on mean arterial pressure (MAP), heart rate (HR), and renal sympathetic nerve activity (RSNA). Glutamate stimulation of PSTN elicited depressor (-20.4 +/- 0.7 mmHg) and bradycardia (-26.0 +/- 1.0 beats/min) responses and decreases in RSNA (67 +/- 17%). Administration (intravenous) of atropine methyl bromide attenuated the bradycardia response (46%), but had no effect on the MAP response. Subsequent intravenous administration of hexamethonium bromide blocked both the remaining bradycardia and depressor responses. Bilateral microinjection of the synaptic blocker CoCl(2) into the caudal NTS region attenuated the PSTN depressor and bradycardia responses by 92% and 94%, respectively. Additionally, prior glutamate activation of neurons in the ipsilateral NTS did not alter the magnitude of the MAP response to stimulation of PSTN, but potentiated HR response by 35%. Finally, PSTN stimulation increased the magnitude of the reflex bradycardia to activation of arterial baroreceptors. These data indicate that activation of neurons in the PSTN elicits a decrease in MAP due to sympathoinhibition and a cardiac slowing that involves both vagal excitation and sympathoinhibition. In addition, these data suggest that the PSTN depressor effects on circulation are mediated in part through activation of NTS neurons involved in baroreflex function. PMID- 18287225 TI - Long-term hypoxia modulates expression of key genes regulating adipose function in the late-gestation ovine fetus. AB - A major function of abdominal adipose in the newborn is nonshivering thermogenesis. Uncoupling protein (UCP) UCP1 and UCP2 play major roles in thermogenesis. The present study tested the hypothesis that long-term hypoxia (LTH) modulates expression of UCP1 and UCP2, and key genes regulating expression of these genes in the late-gestation ovine fetus. Ewes were maintained at high altitude (3,820 m) from 30 to 138 days gestation (dG); perirenal adipose tissue was collected from LTH and age-matched, normoxic control fetuses at 139-141 dG. Quantitative real-time PCR was used to analyze mRNA for UCP1, UCP2, 11beta hydroxysteroid dehydrogenase type 1 (HSD11B1) and 2 (HSD11B2), glucocorticoid receptor (GR), beta3 adrenergic receptor (beta3AR), deiodinase type 1 (DIO1) and DIO2, peroxisome proliferator activated receptor (PPAR) alpha and gamma and PPARgamma coactivator 1 (PGC1alpha). Concentrations of mRNA for UCP1, HSD11B1, PPARgamma, PGC1, DIO1, and DIO2 were significantly higher in perirenal adipose of LTH compared with control fetuses, while mRNA for HSD11B2, GR, or PPARalpha in perirenal adipose did not differ between control and LTH fetuses. The increased expression of UCP1 is likely an adaptive response to LTH, assuring adequate thermogenesis in the event of birth under oxygen-limiting conditions. Because both glucocorticoids and thyroid hormone regulate UCP1 expression, the increase in HSD11B1, DIO1, and DIO2 implicate increased adipose capacity for local synthesis of these hormones. PPARgamma and its coactivator may provide an underlying mechanism via which LTH alters development of the fetal adipocyte. These findings have important implications regarding fetal/neonatal adipose tissue function in response to LTH. PMID- 18287227 TI - Rapid adaptive amplification of preexisting variation in an RNA virus. AB - The amount and nature of preexisting variation in a population of RNA viruses is an important determinant of the virus's ability to adapt rapidly to a changed environment. However, direct quantification of this preexisting variation may be cumbersome, because potentially beneficial alleles are typically rare, and isolation of a large number of subclones is required. Here, we propose a simpler method. We infer the initial population structure of vesicular stomatitis virus (VSV) by fitting a mathematical model of asexual evolution to an extensive set of measurements of VSV fitness dynamics under various conditions, including new and previously published data. The inferred variation of fitness in the initial population agrees very well with the results of direct experiments with subclone fitness quantification. From the same procedure, we also estimate the mean fitness effect of beneficial mutations (selection coefficient s), the percentage of sites in the genome that are under moderate positive or negative selection, and the percentage of sites where beneficial mutations may potentially occur. For VSV strain MARM U evolving in BHK-21 cells, the three parameters have values of 0.39, 9%, and 0.06%, respectively. The method can be generalized and applied easily to other rapidly evolving microbes, including both asexual microorganisms and those with recombination. PMID- 18287226 TI - NF-kappaB-mediated activation of the chemokine CCL22 by the product of the human cytomegalovirus gene UL144 escapes regulation by viral IE86. AB - The product of the human cytomegalovirus (HCMV) gene UL144, expressed at early times postinfection, is located in the UL/b' region of the viral genome and is related to members of the tumor necrosis factor receptor superfamily, but it does not bind tumor necrosis factor superfamily ligands. However, UL144 does activate NF-kappaB, resulting in NF-kappaB-mediated activation of the cellular chemokine CCL22. Consistent with this finding, isolates of HCMV lacking the UL/b' region show no such activation of CCL22. Recently, it has been suggested that activation of NF-kappaB is repressed by the product of the viral gene IE86: IE86 appears to block NF-kappaB binding to DNA but not nuclear translocation of NF-kappaB. Intriguingly, IE86 is detectable throughout an infection with the virus, so how UL144 is able to activate NF-kappaB in the presence of continued IE86 expression is unclear. Here we show that although IE86 does repress the UL144-mediated activation of a synthetic NF-kappaB promoter, it is unable to block UL144 mediated activation of the CCL22 promoter, and this lack of responsiveness to IE86 appears to be regulated by binding of the CREB transcription factor. PMID- 18287228 TI - Theiler's virus-induced intrinsic apoptosis in M1-D macrophages is Bax mediated and restricts virus infectivity: a mechanism for persistence of a cytolytic virus. AB - Theiler's murine encephalomyelitis virus (TMEV), a member of the Cardiovirus genus in the family Picornaviridae, is a highly cytolytic virus that produces necrotic death in rodent cells except for macrophages, which undergo apoptosis. In the present study we have analyzed the kinetics of BeAn virus infection in M1 D cells, in order to temporally relate virus replication to the apoptotic signaling events. Apoptosis was associated with early exponential virus growth from 1 to 12 h postinfection (p.i.); however, >/=80% of peak infectivity was lost by 16 to 24 h p.i. The pan-caspase inhibitor qVD-OPh led to significantly higher virus yields, while zVAD-fmk completely inhibited virus replication until 10 h p.i., precluding its assessment in apoptosis. In contrast, while zVAD-fmk significantly inhibited BeAn virus replication in BHK-21 cells at 12 and 16 h p.i., virus replication at these time points was not altered by qVD-OPh. Bax translocation into mitochondria, efflux of cytochrome c into the cytoplasm, and activation of caspases 9 and 3 between approximately 8 and 12 h p.i. (all hallmarks of the intrinsic apoptotic pathway) were transiently inhibited by expression of Bcl-2, which is not expressed in M1-D cells. Thus, BeAn virus infection in M1-D macrophages, which restricts virus replication, provides a potential mechanism for modulating TMEV neurovirulence during persistence in the mouse central nervous system. PMID- 18287229 TI - Vaccinia virus E2L null mutants exhibit a major reduction in extracellular virion formation and virus spread. AB - The vaccinia virus E2L (VACWR058) gene is conserved in all sequenced chordopoxviruses and is predicted to encode an 86-kDa protein with no recognizable functional motifs or nonpoxvirus homologs. Although the region immediately upstream of the open reading frame lacked optimal consensus promoter motifs, expression of the E2 protein occurred after viral DNA replication. Transfection studies, however, indicated that the promoter was weak compared to well-characterized intermediate and late promoters. The E2 protein was present in mature virions purified from infected cells but was more abundant in extracellular enveloped forms. Despite the conservation of the E2L gene in chordopoxviruses, deletion mutants could be isolated from both the WR and IHD-J strains of vaccinia virus. These null mutants produced very small plaques in all cell lines tested, reduced amounts of mature infectious virions, and very low numbers of extracellular virions. Nevertheless, viral protein synthesis appeared qualitatively and quantitatively normal. The defect in extracellular virus formation was corroborated by electron microscopy, which also showed some aberration in the wrapping of virions by cisternal membranes. Extracellular virions that did form, however, were able to induce actin tail formation. PMID- 18287230 TI - Cytokine responses in porcine respiratory coronavirus-infected pigs treated with corticosteroids as a model for severe acute respiratory syndrome. AB - The effectiveness and potential immunosuppressive effects of anti-inflammatory glucocorticoids in the lungs of severe acute respiratory syndrome (SARS) patients are undefined. We treated porcine respiratory coronavirus (PRCV)-infected conventional pigs with the corticosteroid dexamethasone (DEX) as a model for SARS. Innate and Th1 cytokines in bronchoalveolar lavage (BAL) and serum were elevated in PRCV-infected pigs compared to controls, but were decreased after DEX treatment in the PRCV-infected, DEX-treated (PRCV/DEX) pigs. Although decreased in BAL, Th2 cytokine levels were higher in serum after DEX treatment. Levels of the proinflammatory cytokine interleukin-6 in BAL and serum were decreased in PRCV/DEX pigs early but increased later compared to those in phosphate-buffered saline-treated, PRCV-infected pigs, corresponding to a similar trend for lung lesions. PRCV infection increased T-cell frequencies in BAL, but DEX treatment of PRCV-infected pigs reduced frequencies of T cells; interestingly B and SWC3a(+) (monocytes/macrophages/granulocytes) cell frequencies were increased. DEX reduced numbers of PRCV-stimulated Th1 gamma interferon-secreting cells in spleen, tracheobroncheolar lymph nodes, and blood. Our findings suggest that future glucocorticoid treatment of SARS patients should be reconsidered in the context of potential local immunosuppression of immune responses in lung and systemic Th1 cytokine-biased suppression. PMID- 18287231 TI - Inhibition of CD1 antigen presentation by human cytomegalovirus. AB - The betaherpesvirus human cytomegalovirus (HCMV) encodes several molecules that block antigen presentation by the major histocompatibility complex (MHC) proteins. Humans also possess one other family of antigen-presenting molecules, the CD1 family; however, the effect of HCMV on CD1 expression is unknown. The majority of CD1 molecules are classified on the basis of homology as group 1 CD1 and are present almost exclusively on professional antigen-presenting cells such as dendritic cells, which are a major target for HCMV infection and latency. We have determined that HCMV encodes multiple blocking strategies targeting group 1 CD1 molecules. CD1 transcription is strongly inhibited by the HCMV interleukin-10 homologue cmvIL-10. HCMV also blocks CD1 antigen presentation posttranscriptionally by the inhibition of CD1 localization to the cell surface. This function is not performed by a known HCMV MHC class I-blocking molecule and is substantially stronger than the blockage induced by herpes simplex virus type 1. Antigen presentation by CD1 is important for the development of the antiviral immune response and the generation of mature antigen-presenting cells. HCMV present in antigen-presenting cells thus blunts the immune response by the blockage of CD1 molecules. PMID- 18287232 TI - Alveolar macrophages are a major determinant of early responses to viral lung infection but do not influence subsequent disease development. AB - Macrophages are abundant in the lower respiratory tract. They play a central role in the innate response to infection but may also modulate excessive inflammation. Both macrophages and ciliated epithelial cells respond to infection by releasing soluble mediators, leading to the recruitment of innate and adaptive effector cells. To study the role of lung macrophages in acute respiratory viral infection, we depleted them by the inhalation of clodronate liposomes in an established mouse model of respiratory syncytial virus (RSV) disease. Infection caused an immediate local release of inflammatory cytokines and chemokines, peaking on day 1, which was virtually abolished by clodronate liposome treatment. Macrophage depletion inhibited the activation (days 1 to 2) and recruitment (day 4) of natural killer (NK) cells and enhanced peak viral load in the lung (day 4). However, macrophage depletion did not affect the recruitment of activated CD4 or CD8 T cells, weight loss, or virus-induced changes in lung function. Therefore, lung macrophages play a central role in the early responses to viral infection but have remarkably little effect on the adaptive response occurring at the time of peak disease severity. PMID- 18287233 TI - Identification of a GP64 subdomain involved in receptor binding by budded virions of the baculovirus Autographica californica multicapsid nucleopolyhedrovirus. AB - Enveloped virus entry into host cells is typically initiated by an interaction between a viral envelope glycoprotein and a host cell receptor. For budded virions of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus, the envelope glycoprotein GP64 is involved in host cell receptor binding, and GP64 is sufficient to mediate low-pH-triggered membrane fusion. To better define the role of GP64 in receptor binding, we generated and characterized a panel of antisera against subdomains of GP64. Eight subdomain specific antisera were generated, and their reactivities with GP64 proteins and neutralization of virus infectivity and binding were examined. Antibodies directed against the N-terminal region of GP64 (amino acids 21 to 159) showed strong neutralization of infectivity and effectively inhibited binding of (35)S labeled budded virions to Sf9 cells. In addition, we generated virions displaying truncated GP64 constructs. A construct displaying the N-terminal 274 amino acids (residues 21 to 294) of the ectodomain was sufficient to mediate virion binding. Additional studies of antisera directed against small subdomains revealed that an antiserum against a 40-amino-acid region (residues 121 to 160) neutralized virus infectivity. Site-directed mutagenesis was subsequently used for functional analysis of that region. Recombinant viruses expressing GP64 proteins with single amino acid substitutions within amino acids 120 to 124 and 142 to 148 replicated to high titers, suggesting that those amino acids were not critical for receptor binding or other important GP64 functions. In contrast, GP64 proteins with single amino acid substitutions of residues 153 and 156 were unable to substitute for wild-type GP64 and did not rescue a gp64 knockout virus. Further analysis showed that these substitutions substantially reduced binding of recombinant virus to Sf9 cells. Thus, the amino acid region from positions 21 to 159 was identified as a putative receptor binding domain, and amino acids 153 and 156 appear to be important for receptor binding. PMID- 18287234 TI - Measles virus infects both polarized epithelial and immune cells by using distinctive receptor-binding sites on its hemagglutinin. AB - Measles is one of the most contagious human infectious diseases and remains a major cause of childhood morbidity and mortality worldwide. The signaling lymphocyte activation molecule (SLAM), also called CD150, is a cellular receptor for measles virus (MV), presumably accounting for its tropism for immune cells and its immunosuppressive properties. On the other hand, pathological studies have shown that MV also infects epithelial cells at a later stage of infection, although its mechanism has so far been unknown. In this study, we show that wild type MV can infect and produce syncytia in human polarized epithelial cell lines independently of SLAM and CD46 (a receptor for the vaccine strains of MV). Progeny viral particles are released exclusively from the apical surface of these polarized epithelial cell lines. We have also identified amino acid residues on the MV attachment protein that are likely to interact with a putative receptor on epithelial cells. All of these residues have aromatic side chains and may form a receptor-binding pocket located in a different position from the putative SLAM- and CD46-binding sites on the MV attachment protein. Thus, our results indicate that MV has an intrinsic ability to infect both polarized epithelial and immune cells by using distinctive receptor-binding sites on the attachment protein corresponding to each of their respective receptors. The ability of MV to infect polarized epithelial cells and its exclusive release from the apical surface may facilitate its efficient transmission via aerosol droplets, resulting in its highly contagious nature. PMID- 18287235 TI - Autographa californica multiple nucleopolyhedrovirus nucleocapsid protein BV/ODV C42 mediates the nuclear entry of P78/83. AB - Autographa californica multiple nucleopolyhedrovirus (AcMNPV) BV/ODV-c42 (orf101; c42), which encodes a 41.5-kDa viral nucleocapsid protein with a putative nuclear localization signal (NLS) motif at the C terminus, is a highly conserved gene among members of the Baculoviridae family. C42 is demonstrated to be essential for AcMNPV propagation and can bind to nucleocapsid protein P78/83, a viral activator for the actin-related protein 2/3 (ARP2/3) complex to initiate nuclear actin polymerization, which is essential for viral nucleocapsid morphogenesis during AcMNPV infection. Here, we report the identification of a novel pathway through which c42 functions in nucleocapsid assembly. Cotransfection of Sf9 cells with c42 and p78/83 plasmids demonstrated that C42 was capable of recruiting P78/83 to the nuclei of uninfected cells and that the NLS motif of C42 was essential for this process. To validate this nuclear relocation mode in bacmid transfected cells, a c42-disrupted bacmid (vAc(c42ko-gfp)) and rescued bacmids with wild-type c42 (vAc(c42res-gfp)) or with NLS coding sequence-mutated c42 (vAc(c42nls-gfp)) were prepared. By immuno-staining, P78/83 was found to be localized in the cytoplasm of either vAc(c42ko-gfp)- or vAc(c42nls-gfp) transfected cells, whereas P78/83 was relocated to the nuclei of vAc(c42res-gfp) transfected cells. Furthermore, F-actin-specific staining confirmed that there was no actin polymerization activity in the nuclei of either vAc(c42ko-gfp)- or vAc(c42nls-gfp)-transfected cells, which might be attributed to the absence of nuclear P78/83, an activator of the ARP2/3 complex to initiate nuclear actin polymerization. We therefore hypothesize a mode of action where C42 binds to P78/83 in the cytoplasm to form a protein complex and cotransports to the nucleus under the direction of the NLS motif in C42 during AcMNPV infection. PMID- 18287236 TI - The GLN family of murine endogenous retroviruses contains an element competent for infectious viral particle formation. AB - Several families of endogenous retroviruses (ERVs) have been identified in the mouse genome, in several instances by in silico searches, but for many of them it remains to be determined whether there are elements that can still encode functional retroviral particles. Here, we identify, within the GLN family of highly reiterated ERVs, one, and only one, copy that encodes retroviral particles prone to infection of mouse cells. We show that its envelope protein confers an ecotropic host range and recognizes a receptor different from mCAT1 and mSMIT1, the two previously identified receptors for other ecotropic mouse retroviruses. Electron microscopy disclosed viral particle assembly and budding at the cell membrane, as well as release of mature particles into the extracellular space. These particles are closely related to murine leukemia virus (MLV) particles, with which they have most probably been confused in the past. This study, therefore, identifies a new class of infectious mouse ERVs belonging to the family Gammaretroviridae, with one family member still functional today. This family is in addition to the two MLV and mouse mammary tumor virus families of active mouse ERVs with an extracellular life cycle. PMID- 18287238 TI - XBP-1, a novel human T-lymphotropic virus type 1 (HTLV-1) tax binding protein, activates HTLV-1 basal and tax-activated transcription. AB - X-box binding protein 1 (XBP-1), a basic leucine zipper transcription factor, plays a key role in the cellular unfolded protein response (UPR). There are two XBP-1 isoforms in cells, spliced XBP-1S and unspliced XBP-1U. XBP-1U has been shown to bind to the 21-bp Tax-responsive element of the human T-lymphotropic virus type 1 (HTLV-1) long terminal repeat (LTR) in vitro and transactivate HTLV 1 transcription. Here we identify XBP-1S as a transcription activator of HTLV-1. Compared to XBP-1U, XBP-1S demonstrates stronger activating effects on both basal and Tax-activated HTLV-1 transcription in cells. Our results show that both XBP 1S and XBP-1U interact with Tax and bind to the HTLV-1 LTR in vivo. In addition, elevated mRNA levels of the gene for XBP-1 and several UPR genes were detected in the HTLV-1-infected C10/MJ and MT2 T-cell lines, suggesting that HTLV-1 infection may trigger the UPR in host cells. We also identify Tax as a positive regulator of the expression of the gene for XBP-1. Activation of the UPR by tunicamycin showed no effect on the HTLV-1 LTR, suggesting that HTLV-1 transcription is specifically regulated by XBP-1. Collectively, our study demonstrates a novel host-virus interaction between a cellular factor XBP-1 and transcriptional regulation of HTLV-1. PMID- 18287237 TI - Viral modulation of T-cell receptor signaling. PMID- 18287239 TI - Primer binding site-dependent restriction of murine leukemia virus requires HP1 binding by TRIM28. AB - TRIM28 is a transcriptional corepressor which is required for primer binding site (PBS)-dependent restriction of murine leukemia virus (MLV) replication in embryonic stem and embryonic carcinoma (EC) cells. PBS-dependent restriction of MLV leads to transcriptional silencing of the integrated provirus and has been shown to correlate with TRIM28-mediated recruitment of HP1 to the silenced loci. Here we show, using a cell line with a point mutation in the HP1 binding domain of TRIM28, that interaction with HP1 is absolutely required for the PBS-dependent restriction of MLV in the F9 EC cell line. PMID- 18287240 TI - High-capacity adenovirus vector-mediated anti-glioma gene therapy in the presence of systemic antiadenovirus immunity. AB - Gene therapy is proposed as a novel therapeutic strategy for treating glioblastoma multiforme (GBM), a devastating brain cancer. In the clinic, antivector immune responses pose formidable challenges. Herein we demonstrate that high-capacity adenovirus vectors (HC-Ads) carrying the conditional cytotoxic gene herpes simplex virus type 1-thymidine kinase (TK) induce tumor regression and long-term survival in an intracranial glioma model, even in the presence of systemic antiadenovirus immunity, as could be encountered in patients. First generation Ad-TK failed to elicit tumor regression in this model. These results pave the way for implementing HC-Ad-TK-mediated gene therapy as a powerful adjuvant for treating GBM. PMID- 18287241 TI - Cell surface expression of the vaccinia virus complement control protein is mediated by interaction with the viral A56 protein and protects infected cells from complement attack. AB - The vaccinia virus (VACV) complement control protein (VCP) is the major protein secreted from VACV-infected cells. It has been reported that VCP binds to the surfaces of uninfected cells by interacting with heparan sulfate proteoglycans (HSPGs). In this study, we show that VCP is also expressed on the surfaces of infected cells and demonstrate that surface localization occurs independently of HSPGs. Since VCP does not contain a transmembrane domain, we hypothesized that VCP interacts with a membrane protein that localizes to the infected-cell surface. We show that the VACV A56 membrane protein is necessary for the cell surface expression of VCP and demonstrate that VCP and A56 interact in VACV infected cells. Since the surface expression of VCP was abrogated by reducing agents, we examined the contribution of an unpaired cysteine residue on VCP to VCP surface expression and VCP's interaction with A56. To do this, we mutated the unpaired cysteine in VCP and generated a recombinant virus expressing the altered form of VCP. Following the infection of cells with the mutant virus, VCP was neither expressed on the cell surface nor able to interact with A56. Importantly, the cell surface expression of VCP was found to protect infected cells from complement-mediated lysis. Our findings suggest a new function for VCP that may be important for poxvirus pathogenesis and impact immune responses to VACV-based vaccines. PMID- 18287242 TI - Calibration of multiple poliovirus molecular clocks covering an extended evolutionary range. AB - We have calibrated five different molecular clocks for circulating poliovirus based upon the rates of fixation of total substitutions (K(t)), synonymous substitutions (K(s)), synonymous transitions (A(s)), synonymous transversions (B(s)), and nonsynonymous substitutions (K(a)) into the P1/capsid region (2,643 nucleotides). Rates were determined over a 10-year period by analysis of sequences of 31 wild poliovirus type 1 isolates representing a well-defined phylogeny derived from a common imported ancestor. Similar rates were obtained by linear regression, the maximum likelihood/single-rate dated-tip method, and Bayesian inference. The very rapid K(t) [(1.03 +/- 0.10) x 10(-2) substitutions/site/year] and K(s) [(1.00 +/- 0.08) x 10(-2)] clocks were driven primarily by the A(s) clock [(0.96 +/- 0.09) x 10(-2)], the B(s) clock was approximately 10-fold slower [(0.10 +/- 0.03) x 10(-2)], and the more stochastic K(a) clock was approximately 30-fold slower [(0.03 +/- 0.01) x 10(-2)]. Nonsynonymous substitutions at all P1/capsid sites, including the neutralizing antigenic sites, appeared to be constrained by purifying selection. Simulation of the evolution of third-codon positions suggested that saturation of synonymous transitions would be evident at 10 years and complete at approximately 65 years of independent transmission. Saturation of synonymous transversions was predicted to be minimal at 20 years and incomplete at 100 years. The rapid evolution of the K(t), K(s), and A(s) clocks can be used to estimate the dates of divergence of closely related viruses, whereas the slower B(s) and K(a) clocks may be used to explore deeper evolutionary relationships within and across poliovirus genotypes. PMID- 18287243 TI - Proteomic studies reveal coordinated changes in T-cell expression patterns upon infection with human immunodeficiency virus type 1. AB - We performed an extensive two-dimensional differential in-gel electrophoresis proteomic analysis of the cellular changes in human T cells upon human immunodeficiency virus type 1 (HIV-1) infection. We detected 2,000 protein spots, 15% of which were differentially expressed at peak infection. A total of 93 proteins that changed in relative abundance were identified. Of these, 27 were found to be significantly downregulated and 66 were upregulated at peak HIV infection. Early in infection, only a small group of proteins was changed. A clear and consistent program of metabolic rerouting could be seen, in which glycolysis was downregulated and mitochondrial oxidation enhanced. Proteins that participate in apoptotic signaling were also significantly influenced. Apart from these changes, the virus also strongly influenced levels of proteins involved in intracellular transport. These and other results are discussed in light of previous microarray and proteomic studies regarding the impact of HIV-1 infection on cellular mRNA and protein content. PMID- 18287244 TI - Adenovirus E3/19K promotes evasion of NK cell recognition by intracellular sequestration of the NKG2D ligands major histocompatibility complex class I chain related proteins A and B. AB - The adenovirus (Ad) early transcription unit 3 (E3) encodes multiple immunosubversive functions that are presumed to facilitate the establishment and persistence of infection. Indeed, the capacity of E3/19K to inhibit transport of HLA class I (HLA-I) to the cell surface, thereby preventing peptide presentation to CD8(+) T cells, has long been recognized as a paradigm for viral immune evasion. However, HLA-I downregulation has the potential to render Ad-infected cells vulnerable to natural killer (NK) cell recognition. Furthermore, expression of the immediate-early Ad gene E1A is associated with efficient induction of ligands for the key NK cell-activating receptor NKG2D. Here we show that while infection with wild-type Ad enhances synthesis of the NKG2D ligands, major histocompatibility complex class I chain-related proteins A and B (MICA and MICB), their expression on the cell surface is actively suppressed. Both MICA and MICB are retained within the endoplasmic reticulum as immature endoglycosidase H sensitive forms. By analyzing a range of cell lines and viruses carrying mutated versions of the E3 gene region, E3/19K was identified as the gene responsible for this activity. The structural requirements within E3/19K necessary to sequester MICA/B and HLA-I are similar. In functional assays, deletion of E3/19K rendered Ad-infected cells more sensitive to NK cell recognition. We report the first NK evasion function in the Adenoviridae and describe a novel function for E3/19K. Thus, E3/19K has a dual function: inhibition of T-cell recognition and NK cell activation. PMID- 18287245 TI - Alveolar macrophages are indispensable for controlling influenza viruses in lungs of pigs. AB - Alveolar macrophages constitutively reside in the respiratory tracts of pigs and humans. An in vivo role of alveolar macrophages in defending against influenza viruses in mice infected with a reassorted influenza virus, 1918 HA/NA:Tx/91, was reported, but there has been no report on an in vivo role of alveolar macrophages in a natural host such as a pig using currently circulating human influenza virus. Here we show that in vivo depletion of alveolar macrophages in pigs by dichloromethylene diphosphonate (MDPCL2) treatment results in 40% mortality when pigs are infected with currently circulating human H1N1 influenza viruses, while none of the infected control pigs died. All infected pigs depleted of alveolar macrophages suffered from more severe respiratory signs than infected control pigs. Induction of tumor necrosis factor alpha in the infected pigs depleted of alveolar macrophages was significantly lower than that in the lungs of infected control pigs, and the induction of interleukin-10, an immunosuppressive cytokine, significantly increased in the lungs of infected pigs depleted of alveolar macrophages compared to infected control pigs. When we measured antibody titers and CD8(+) T lymphocytes expressing gamma interferon (IFN-gamma), lower antibody titers and a lower percentage of CD8(+) T lymphocytes expressing IFN-gamma were detectable in MDPCL2-treated infected pigs than in phosphate-buffered saline- and liposome-treated and infected pigs. Taken together, our findings suggest that alveolar macrophages are essential for controlling H1N1 influenza viruses in pigs. PMID- 18287246 TI - Deletion of Epstein-Barr virus BFLF2 leads to impaired viral DNA packaging and primary egress as well as to the production of defective viral particles. AB - Previous genetic and biochemical studies performed with several members of the Alphaherpesvirus subfamily have shown that the UL31 and UL34 proteins are essential components of the molecular machinery that mediates the primary egress of newly assembled capsids across the nuclear membrane. Further, there is substantial evidence that BFLF2 and BFRF1, the respective positional homologs of UL31 and UL34 in the Epstein-Barr virus (EBV) genome, are also their functional homologs, i.e., that the UL31/UL34 pathway is common to distant herpesviruses. However, the low degree of protein sequence identity between UL31 and BFLF2 would argue against such a hypothesis. To further clarify this issue, we have constructed a recombinant EBV strain devoid of BFLF2 (DeltaBFLF2) and show that BFLF2 is crucial for efficient virus production but not for lytic DNA replication or B-cell transformation. This defective phenotype could be efficiently restored by trans complementation with a BFLF2 expression plasmid. Detailed analysis of replicating cells by electron microscopy revealed that, as expected, DeltaBFLF2 viruses not only failed to egress from the nucleus but also showed defective DNA packaging. Nonfunctional primary egress did not, however, impair the production and extracellular release of enveloped but empty viral particles that comprised L particles containing tegument-like structures and a few virus-like particles carrying empty capsids. The DeltaBFLF2 and DeltaUL31 phenotypes therefore only partly overlap, from which we infer that BFLF2 and UL31 have substantially diverged during evolution to fulfil related but distinct functions. PMID- 18287247 TI - Regulation of Smad-mediated gene transcription by RGS3. AB - Regulator of G protein signaling (RGS) proteins are united into a family by the presence of the homologous RGS domain that binds the alpha subunits of heterotrimeric G proteins and accelerates their GTPase activity. A member of this family, RGS3 regulates the signaling mediated by G(q) and G(i) proteins by binding the corresponding Galpha subunits. Here we show that RGS3 interacts with the novel partners Smad2, Smad3, and Smad4-the transcription factors that are activated through a transforming growth factor-beta (TGF-beta) receptor signaling. This interaction is mediated by the region of RGS3 outside of the RGS domain and by Smad's Mad homology 2 domain. Overexpression of RGS3 results in inhibition of Smad-mediated gene transcription. RGS3 does not affect TGF-beta induced Smad phosphorylation, but it prevents heteromerization of Smad3 with Smad4, which is required for transcriptional activity of Smads. This translates to functional inhibition of TGF-beta-induced myofibroblast differentiation by RGS3. In conclusion, this study identifies a novel, noncanonical role of RGS3 in regulation of TGF-beta signaling through its interaction with Smads and interfering with Smad heteromerization. PMID- 18287248 TI - Flavopiridol suppresses tumor necrosis factor-induced activation of activator protein-1, c-Jun N-terminal kinase, p38 mitogen-activated protein kinase (MAPK), p44/p42 MAPK, and Akt, inhibits expression of antiapoptotic gene products, and enhances apoptosis through cytochrome c release and caspase activation in human myeloid cells. AB - Although flavopiridol, a semisynthetic flavone, was initially thought to be a specific inhibitor of cyclin-dependent kinases, it has now been shown that flavopiridol mediates antitumor responses through mechanism(s) yet to be defined. We have shown previously that flavopiridol abrogates tumor necrosis factor (TNF) induced nuclear factor-kappaB (NF-kappaB) activation. In this report, we examined whether this flavone affects other cellular responses activated by TNF. TNF is a potent inducer of activator protein-1 (AP-1), and flavopiridol abrogated this activation in a dose- and time-dependent manner. Flavopiridol also suppressed AP 1 activation induced by various carcinogens and inflammatory stimuli. When examined for its effect on other signaling pathways, flavopiridol inhibited TNF induced activation of various mitogen-activated protein kinases, including c-Jun NH(2)-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), and p44/p42 MAPK. It is noteworthy that this flavone also suppressed TNF-induced activation of Akt, a cell survival kinase, and expression of various antiapoptotic proteins, such as IAP-1, IAP-2, XIAP, Bcl-2, Bcl-xL, and TRAF-1. Flavopiridol also inhibited the TNF-induced induction of intercellular adhesion molecule-1, c-Myc, and c-Fos, all known to mediate tumorigenesis. Moreover, TNF induced apoptosis was enhanced by flavopiridol through activation of the bid cytochrome-caspase-9-caspase-3 pathway. Overall, our results clearly suggest that flavopiridol interferes with the TNF cell-signaling pathway, leading to suppression of antiapoptotic mechanisms and enhancement of apoptosis. PMID- 18287249 TI - Risk for nephrogenic systemic fibrosis with gadoteridol (ProHance) in patients who are on long-term hemodialysis. AB - BACKGROUND AND OBJECTIVES: Recent studies strongly link nephrogenic systemic fibrosis to gadolinium administration for magnetic resonance imaging. In a recent advisory, the Food and Drug Administration stated that all gadolinium-containing chelates are potentially associated with nephrogenic systemic fibrosis; however, most reported cases are linked to gadodiamide (Omniscan) and gadopentetate dimeglumine (Magnevist). Given the severe consequences of nephrogenic systemic fibrosis, it is critical to define the risks associated with each gadolinium containing chelate. The purpose of this study was to examine nephrogenic systemic fibrosis risk in a hemodialysis population exposed to gadoteridol (ProHance). DESIGN, SETTING, PARTICIPANTS, & OBJECTIVES: Appointment logs were used to generate a database of all long-term hemodialysis patients at the Dallas Veterans Affairs hospital since August 2001. These patients were then examined in the Veterans Affair's electronic medical record system for gadolinium exposure during magnetic resonance imaging from 2000 through 2007, a period during which gadoteridol was the sole contrast agent used. RESULTS: A total of 141 patients were identified with 198 gadoteridol exposures. No cases of nephrogenic systemic fibrosis were identified. The observed frequency of nephrogenic systemic fibrosis was compared with the expected frequency (2.4%) using one-way chi(2) and binomial analysis, yielding a P < 0.05, indicating that the result was not explained by chance alone. CONCLUSIONS: It is concluded that the risk for nephrogenic systemic fibrosis with gadoteridol in patients who are on long-term hemodialysis may be lower than with gadodiamide and gadopentetate dimeglumine. PMID- 18287250 TI - Management of cardiovascular disease in renal transplant recipients. AB - Cardiovascular disease is a major cause of graft loss and the leading cause of death in renal transplant recipients. Although there are robust data on the frequency of risk factors and their contributions to cardiovascular disease in this population, few trials have demonstrated the benefit of modifying these risk factors to reduce cardiovascular events. Nevertheless, it is widely accepted that the clinical acumen filtered through the best available studies in the general population be used to treat individual renal transplant recipients given their high cardiovascular mortality. Transplant task forces and the Kidney Disease Outcomes Quality Initiative have created guidelines for this purpose. This review examines the data available for prevention and treatment of major risk factors contributing to cardiovascular disease in renal transplant recipients. The contribution of immunosuppressive agents to each risk factor and the evidence to support lifestyle modification as well as drug therapy are examined. Reducing cardiovascular risk factors requires an integrative approach that is best accomplished by a team of health care professionals. It creates a significant challenge but one that must be met if allograft survival is to improve. PMID- 18287251 TI - A framework and key research questions in AKI diagnosis and staging in different environments. AB - BACKGROUND AND OBJECTIVES: Acute Kidney Injury (AKI) is common worldwide, and associated with significant morbidity, mortality, and resource utilization. The RIFLE system of staging AKI correlates with survival in AKI in several settings. A similar AKI definition and staging system that also incorporates lesser degrees of serum creatinine elevation was proposed at the inaugural Acute Kidney Injury Network (AKIN) meeting in 2005. At the Second AKIN meeting in Vancouver, Canada in September 2006, our group developed a research agenda that would test the utility of these diagnostic and staging criteria to predict patient outcomes in a variety of clinical settings and patient groups. DESIGN, SETTING, PARTICIPANTS & MEASUREMENTS: Three-day, international, consensus conference. A multidisciplinary stakeholder committee was divided into work groups. Recommendations for clinical practice and for future research were developed by the committee as an iterative process. This procedure consisted of a literature review phase and focus group interactions with presentations to the entire committee. RESULTS: We first proposed a conceptual framework of disease that describes a series of AKI stages, antecedents and outcomes, and allows a description of research recommendations based on transition between AKI stages. We further proposed methods for testing of the definition and development of research questions to establish the utility of new biomarkers for the diagnosis and staging of AKI and associated illnesses. CONCLUSIONS: Retrospective studies should be conducted to initiate the process of validating the AKIN definition of AKI, followed by comprehensive prospective studies that incorporate sampling for emerging AKI biomarkers. PMID- 18287252 TI - Necrotizing and crescentic lupus nephritis with antineutrophil cytoplasmic antibody seropositivity. AB - BACKGROUND AND OBJECTIVES: Lupus nephritis is a classic immune complex glomerulonephritis. In contrast, antineutrophil cytoplasmic antibodies are associated with necrotizing and crescentic glomerulonephritis, in the absence of significant immune deposits. Antineutrophil cytoplasmic antibodies are detected by indirect immunofluorescence in 20% of patients with systemic lupus erythematosus. We report 10 cases of necrotizing and crescentic lupus nephritis with antineutrophil cytoplasmic antibody seropositivity. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Ten patients with systemic lupus erythematosus, antineutrophil cytoplasmic antibody positivity, and renal biopsy findings of lupus nephritis and antineutrophil cytoplasmic antibody-associated glomerulonephritis were identified. The clinical features, pathologic findings, and outcomes are described. RESULTS: The cohort consisted of eight women and two men with a mean age of 48.4 yr. Perinuclear antineutrophil cytoplasmic antibody was detected by indirect immunofluorescence in nine patients. Four of the nine patients and the single remaining patient were found to have myeloperoxidase antineutrophil cytoplasmic antibodies by enzyme-linked immunosorbent assay. Clinical presentation included proteinuria, hematuria, and acute renal insufficiency, with mean creatinine of 7.1 mg/dl. All biopsies exhibited prominent necrosis and crescents with absent or rare subendothelial deposits and were interpreted as lupus nephritis and antineutrophil cytoplasmic antibody associated glomerulonephritis. All patients received cyclophosphamide and prednisone. Three patients died of infectious complications. Among the remaining seven patients, five achieved a complete or near-complete remission, one had a remission with subsequent relapse, and one had no response to therapy. CONCLUSION: Antineutrophil cytoplasmic antibody-associated necrotizing and crescentic glomerulonephritis may occur superimposed on lupus nephritis. In patients with lupus nephritis and biopsy findings of prominent necrosis and crescent formation in the absence of significant endocapillary proliferation or subendothelial deposits, antineutrophil cytoplasmic antibody testing by enzyme linked immunosorbent assay is recommended. PMID- 18287253 TI - Screening for chronic kidney disease: where does Europe go? AB - This review discusses various screening approaches for chronic kidney disease that are used in Europe. The criterion for defining chronic kidney disease in the various programs differs but is frequently limited to estimated glomerular filtration rate, thus offering only data on chronic kidney disease stages 3 and higher; however, screening should not be limited to measuring only estimated glomerular filtration rate but should also include a measure of microalbuminuria, because this will offer identification of chronic kidney disease stages 1 and 2. Defining these earlier stages is of importance because the risk for developing end-stage renal disease that is associated with stages 1 and 2 is nearly equal to the risk that is associated with stage 3. Moreover, the risk for cardiovascular events in stages 1 and 2 is equal to that in stage 3. Various reports argue that costs of screening programs in general practitioner or outpatient offices are high and that they are cost-effective only for preventing end-stage renal disease when they are limited to target groups, such as patients with diabetes or hypertension and elderly. The benefits of screening programs, however, should not be evaluated only with respect to the prevention of renal events but should also include the benefits of preventing cardiovascular events. The use of preselection based on either an impaired estimated glomerular filtration rate or on protein dipstick positivity or elevated albuminuria in a morning urine void has been found effective in various European countries as an alternative for targeted screening. PMID- 18287254 TI - Serum albumin level and risk for mortality and hospitalization in adolescents on hemodialysis. AB - BACKGROUND AND OBJECTIVES: National Kidney Foundation Dialysis Outcomes Quality Initiative practice guidelines recommend serum albumin > or = 4.0 g/dl for adults who are on hemodialysis. There is no established pediatric target for albumin and little evidence to support use of adult guidelines. This study examined the association between albumin and risk for death and hospitalization in adolescents who are on hemodialysis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This retrospective cohort study linked data on patients aged 12 to 18 yr in 1999 and 2000 from the Centers for Medicare and Medicaid Services' End Stage Renal Disease Clinical Performance Measures Project with 4-yr hospitalization and mortality records in the United States Renal Data System. Albumin was categorized as < 3.5/3.2, > or = 3.5/3.2 and < 4.0/3.7, and > or = 4.0/3.7 g/dl. RESULTS: Of 675 adolescents, 557 were hospitalized and 50 died. Albumin > or = 4.0/3.7 g/dl was associated with male gender, Hispanic ethnicity, and higher hemoglobin level. Those with albumin > or = 4.0/3.7 g/dl had fewer deaths per 100 patient-years and fewer hospitalizations per time at risk. In multivariate analysis, patients with albumin > or = 4.0/3.7 g/dl had 57% decreased risk for death. Poisson regression showed progressive decrease in hospitalization risk as albumin level increased; however, confidence intervals were similar between albumin > or = 4.0/3.7 g/dl and albumin > or = 3.5/3.2 and < 4.0/3.7 g/dl. CONCLUSIONS: This study demonstrates decreased mortality and hospitalization risk with albumin > or = 3.5/3.2 g/dl and suggests that adolescent hemodialysis patients who are able to achieve serum albumin > or = 4.0/3.7 g/dl may have the lowest mortality risk. PMID- 18287255 TI - C.E.R.A. corrects anemia in patients with chronic kidney disease not on dialysis: results of a randomized clinical trial. AB - BACKGROUND AND OBJECTIVES: This study examined the efficacy of C.E.R.A., a continuous erythropoietin receptor activator, for correcting anemia in patients who had chronic kidney disease (CKD) and were not on dialysis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In this open-label, randomized, parallel-group, Phase III study, 324 adult patients with CKD not on dialysis nor receiving treatment with erythropoiesis-stimulating agents (ESAs) were randomly assigned (1:1) to receive subcutaneous C.E.R.A. once every 2 wk or darbepoetin alfa once weekly during an 18-wk correction period and a 10-wk evaluation period. Thereafter, patients receiving C.E.R.A. were randomly assigned to C.E.R.A. once every 2 wk or once monthly, and patients receiving darbepoetin alfa could receive darbepoetin alfa once weekly or once every 2 wk for a 24-wk extension period. Dosage was adjusted to achieve a hemoglobin (Hb) response and to maintain Hb +/-1 g/dl of the response level and 11 to 13 g/dl. Primary end points were Hb response rate during correction and evaluation and change in Hb concentration between baseline and evaluation. RESULTS: Hb response rates were 97.5% for C.E.R.A. and 96.3% for darbepoetin alfa. Adjusted mean changes in Hb from baseline to evaluation were 2.15 g/dl (C.E.R.A.) and 2.00 g/dl (darbepoetin alfa). Analysis showed that C.E.R.A. once every 2 wk was as effective as darbepoetin alfa once weekly for correcting anemia. Hb levels remained stable in all groups during the extension period. C.E.R.A. and darbepoetin alfa were well tolerated. CONCLUSIONS: Subcutaneous C.E.R.A. once every 2 wk corrects anemia in ESA-naive patients who are not on dialysis. PMID- 18287256 TI - Racial disparities in chronic kidney disease: tragedy, opportunity, or both? PMID- 18287257 TI - Pregnancy in women on dialysis: is success a matter of time? PMID- 18287258 TI - What have we learned about optimal induction therapy for lupus nephritis (III through V) from randomized, controlled trials? PMID- 18287259 TI - Superovulation alters the expression of imprinted genes in the midgestation mouse placenta. AB - Imprinted genes play important roles in embryonic growth and development as well as in placental function. Many imprinted genes acquire their epigenetic marks during oocyte growth, and this period may be susceptible to epigenetic disruption following hormonal stimulation. Superovulation has been shown to affect growth and development of the embryo, but an effect on imprinted genes has not been shown in postimplantation embryos. In the present study, we examined the effect of superovulation/in vivo development or superovulation/3.5dpc (days post-coitum) embryo transfer on the allelic expression of Snrpn, Kcnq1ot1 and H19 in embryos and placentas at 9.5 days of gestation. Superovulation followed by in vivo development resulted in biallelic expression of Snrpn and H19 in 9.5dpc placentas while Kcnq1ot1 was not affected; in the embryos, there was normal monoallelic expression of the three imprinted genes. We did not observe significant DNA methylation perturbations in the differentially methylated regions of Snrpn or H19. Superovulation followed by embryo transfer at 3.5dpc resulted in biallelic expression of H19 in the placenta. The expression of an important growth factor closely linked to H19, Insulin-like growth factor-II, was increased in the placenta following superovulation with or without embryo transfer. These results show that both maternally and paternally methylated imprinted genes were affected, suggesting that superovulation compromises oocyte quality and interferes with the maintenance of imprinting during preimplantation development. Our findings contribute to the evidence that mechanisms for maintaining imprinting are less robust in trophectoderm-derived tissues, and have clinical implications for the screening of patients following assisted reproduction. PMID- 18287260 TI - 1-11C-methyl-4-piperidinyl-N-butyrate radiation dosimetry in humans by dynamic organ-specific evaluation. AB - Deficits of cholinergic neurotransmission contribute to various neurologic and psychiatric conditions. The neurotransmitter acetylcholine is hydrolyzed in the synaptic clefts by 2 enzymes, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). 1-[(11)C]-Methyl-4-piperidinyl-N-butyrate ((11)C MP4B) is a radioligand for the assessment study of BuChE activity in human brain with PET. In the present study the radiation-absorbed doses of the (11)C-MP4B were estimated in humans according to the guidelines of the International Commission on Radiological Protection. Two different data acquisition protocols dynamic organ-specific evaluation (DOSE) and whole-body scanning-were compared. Both methods are widely used for evaluation of radiation burden of (11)C-labeled PET tracers. METHODS: Fixed-bed PET on the upper neck, thorax, abdomen, or pelvic region was performed on 7 healthy subjects after injection of 707 +/- 34 MBq (mean +/- SD) of (11)C-MP4B. Brain input was derived from our previous studies on 18 healthy control subjects and 10 patients with Alzheimer's disease. Regions of interest were drawn on transverse images of all visible organs. Radiation dose estimates were calculated from organ residence times using the MIRDOSE3 software. Urine samples were collected after imaging to estimate tracer extraction. To compare the estimates for absorbed doses between the whole-body scan approach and the DOSE method, we simulated whole-body data acquisition methods used in (11)C dosimetry studies with our fixed-bed data. RESULTS: The organs with the highest radiation-absorbed doses were the liver, urinary bladder, kidneys (renal cortex), upper large intestine, trabecular bone, salivary glands, and heart wall. Up to 60% of the injected dose was excreted via the urinary pathway, and the clearance was relatively rapid, as 30% of the radioactivity was excreted within 60 min after injection. With a 2-h voiding interval the effective dose was 4.2 microSv/MBq. CONCLUSION: (11)C-MP4B causes less radiation burden than previously studied (11)C-labeled PET tracers. No intolerably high absorbed doses were observed in critical organs. With 740 MBq of injected radioactivity, the radiation burden is equivalent to 3.11 mSv. This would allow multiple PET examinations per year to be performed on the same subject. The DOSE method and the simulated whole-body imaging approach produced similar results. PMID- 18287262 TI - Molecular imaging of bcl-2 expression in small lymphocytic lymphoma using 111In labeled PNA-peptide conjugates. AB - The bcl-2 gene is overexpressed in non-Hodgkin's lymphoma (NHL), such as small lymphocytic lymphoma (SLL), and many other cancers. Noninvasive imaging of bcl-2 expression has the potential to identify patients at risk for relapse or treatment failure. The purpose of this study was to synthesize and evaluate radiolabeled peptide nucleic acid (PNA)-peptide conjugates targeting bcl-2 gene expression. An (111)In-labeled PNA complementary to the translational start site of bcl-2 messenger RNA was attached to Tyr(3)-octreotate for somatostatin receptor-mediated intracellular delivery. METHODS: DOTA-anti-bcl-2-PNA-Tyr(3) octreotate (1) and 3 control conjugates (DOTA-nonsense-PNA-Tyr(3)-octreotate (2), DOTA-anti-bcl-2-PNA-Ala[3,4,5,6]-substituted congener (3), and DOTA-Tyr(3) octreotate (4) [DOTA is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid]) were synthesized by standard solid-phase 9-fluorenylmethoxycarbonyl (Fmoc) chemistry. In vitro studies were performed in Mec-1 SLL cells, which express both bcl-2 messenger RNA and somatostatin receptors. Biodistributions and microSPECT/CT studies were performed in Mec-1-bearing SCID (severe combined immunodeficiency) mice, a new animal model of human SLL. RESULTS: (111)In-Labeled conjugate 1 was taken up by Mec-1 cells through a somatostatin receptor-mediated mechanism. Biodistribution studies showed specific tumor uptake of conjugate 1, the somatostatin analog 4, and the PNA nonsense conjugate 2, but not of the mutant peptide conjugate 3. Mec-1 tumors could be detected by microSPECT/CT using (111)In-labeled DOTA-Tyr(3)-octreotate (4) and the targeted anti-bcl-2 conjugate (1), but not using the 2 negative control conjugates 2 and 3. CONCLUSION: A new (111)In-labeled antisense PNA-peptide conjugate demonstrated proof of principle for molecular imaging of bcl-2 expression in a new mouse model of human SLL. This imaging agent may be useful for identifying NHL patients at risk for relapse and conventional treatment failure. PMID- 18287261 TI - Characterization of osteolytic, osteoblastic, and mixed lesions in a prostate cancer mouse model using 18F-FDG and 18F-fluoride PET/CT. AB - The combination of small-animal PET/CT scans and conventional imaging methods may enhance the evaluation of in vivo biologic interactions of murine models in the study of prostate cancer metastasis to bone. METHODS: Small-animal PET/CT scans using (18)F-fluoride ion and (18)F-FDG coregistered with high-resolution small animal CT scans were used to longitudinally assess the formation of osteoblastic, osteolytic, and mixed lesions formed by human prostate cancer cell lines in a severe combined immunodeficient (SCID) mouse tibial injection model. These scans were correlated with plain radiographs, histomorphometry, and soft-tissue measurements. RESULTS: Small-animal PET/CT scans were able to detect biologic activity of cells that induced an osteoblastic lesion 2 wk earlier than on plain radiographs. Furthermore, both the size and the activity of the lesions detected on PET/CT images significantly increased at each successive time point (P < 0.05). (18)F-FDG lesions strongly correlated with soft-tissue measurements, whereas (18)F-fluoride ion activity correlated with bone volume measured on histomorphometric analysis (P < 0.005). Osteolytic lesions were successfully quantified using small-animal CT, whereas lesion sizes measured on (18)F-FDG PET scans also strongly correlated with soft-tissue tumor burden (P < 0.05). In contrast, for mixed lesions, (18)F-fluoride ion and (18)F-FDG PET/CT scans detected only minimal activity. CONCLUSION: (18)F-FDG and (18)F-fluoride ion PET/CT scans can be useful tools in characterizing pure osteolytic and osteoblastic lesions induced by human prostate cancer cell lines. The value of this technology needs further evaluation to determine whether these studies can be used effectively to detect more subtle responses to different treatment regimens in animal models. PMID- 18287263 TI - Staging pathways in recurrent colorectal carcinoma: is contrast-enhanced 18F-FDG PET/CT the diagnostic tool of choice? AB - (18)F-FDG PET/CT has gained wide acceptance for evaluation of recurrent colorectal carcinoma. However in clinical practice, contrast-enhanced CT (ceCT) is still the first-line restaging tool. The aim of this study was to investigate the value of contrast-enhanced PET/CT (cePET/CT) as a first-line restaging tool with a special focus on the importance of the use of intravenous contrast. METHODS: Fifty-four patients (17 women, 37 men; mean age, 60.3 y), referred for restaging of colorectal carcinoma, were examined with cePET/CT. Retrospective analysis was performed by 2 experienced readers by consensus: first, ceCT alone; second, non-cePET/CT; and third, cePET/CT. The number, localization, and diagnostic certainty of lesions were evaluated. Additionally, the therapeutic impact of the findings was determined. In 29 patients, histology, clinical imaging, and clinical follow-up served as the reference standard. In 25 patients, clinical follow-up and imaging served as the reference standard. RESULTS: Overall, non-cePET/CT delivered correct additional information to the ceCT findings in 27 of 54 patients (50%). This occurred in (a) 20 of 30 patients, where ceCT was found to be inconclusive, and in (b) 7 of 24 patients with conclusive ceCT findings, where non-cePET/CT found additional lesions, leading to a therapy modification in 5 patients. Compared with non-cePET/CT, cePET/CT revealed additional information in 39 of 54 patients (72%), with therapeutic relevance in 23 patients. This large number was primarily due to correct segmental localization of liver metastases, which is crucial for surgical therapy planning. CONCLUSION: On the basis of its higher accuracy and therapeutic impact compared with ceCT, our data suggest that cePET/CT might be considered as the first-line diagnostic tool for restaging in patients with colorectal cancer. PMID- 18287264 TI - Canine anatomic phantom for preclinical dosimetry in internal emitter therapy. AB - The majority of investigational studies of new diagnostic and therapeutic radiopharmaceuticals use murine animal models for preclinical assessments of pharmacokinetics and organ radiation dosimetry. Although mice and rats are widely available and relatively inexpensive, their smaller organ anatomy relative to that of humans can lead to considerable differences in organ dosimetry, thus complicating extrapolations of dose-response relationships to human patients. Nonhuman primates circumvent these problems in many respects but are increasingly becoming expensive and limited because of ethical considerations. With the recent completion of the dog genome project and the recognition of many similarities between canine and human cancers, dogs are increasingly being considered in cancer research and drug development. The main objective of this study was to construct a 3-dimensional computational phantom of a large dog on the basis of whole-body multislice CT data. METHODS: A female hound cross underwent whole-body contrast-enhanced CT at a 2-mm slice thickness. On completion of the scan, the dog was euthanized, and the entire skeleton was harvested for a subsequent microCT investigation. The CT data were imported into a computational software program and used to create a polygon-mesh phantom of the entire animal. All of the major organs and bones were semiautomatically segmented and tagged to the CT slices. The phantom data were imported into a second software program and transformed to a nonuniform rational basis-spline surface phantom, allowing easy alteration of the phantom to simulate dogs of smaller or larger statures. A voxel based version of the canine phantom was created by use of an in-house routine for subsequent import into the EGSnrc radiation transport code for photon and beta particle organ dosimetry. RESULTS: The resulting voxel-based version of the canine phantom had a total body mass of 26.0 kg and a total body tissue mass (exclusive of wall organ content) of 24.5 kg. Although this University of Florida (UF) canine phantom displayed a total body mass intermediate between those of the Oak Ridge National Laboratory (ORNL) 5-y and 10-y stylized human phantoms of the MIRDOSE and OLINDA software codes, considerable differences were noted in organ photon cross-doses. For example, ratios of the specific absorbed fraction Phi(lungs <-- liver)(UF Dog) to Phi(lungs <-- liver)(ORNL 5-y) ranged from approximately 30 at 10 keV to approximately 3.5 at 1 MeV. Corresponding ratios of Phi(lungs <-- liver)(UF Dog) to Phi(lungs <-- liver)(ORNL 10-y) ranged from approximately 6 at 10 keV to approximately 1.3 at 1 MeV. Conversely, values of Phi(kidneys <-- spleen) and Phi(liver <-- spleen) were noted to be much lower (factors of 2-4) and much higher (factors of 2-15), respectively, in the canine phantom than in the ORNL human phantoms. These differences were attributed more to organ shape and position within the torso than to organ mass, because many of the canine organs closely approximated their counterparts volumetrically in the stylized pediatric human phantoms. CONCLUSION: The use of canine models, particularly in spontaneously occurring malignancies such as osteosarcoma, for preclinical testing of antineoplastic agents offers significant advantages over current murine models. However, the development of canine-specific technology is critical to the optimization of these studies. The UF canine dosimetry phantom described here aims to solve problems that could stem from the use of current human dosimetry models during radiopharmaceutical research. PMID- 18287265 TI - Dynamic small-animal PET imaging of tumor proliferation with 3'-deoxy-3'-18F fluorothymidine in a genetically engineered mouse model of high-grade gliomas. AB - 3'-Deoxy-3'-(18)F-fluorothymidine ((18)F-FLT), a partially metabolized thymidine analog, has been used in preclinical and clinical settings for the diagnostic evaluation and therapeutic monitoring of tumor proliferation status. We investigated the use of (18)F-FLT for detecting and characterizing genetically engineered mouse (GEM) high-grade gliomas and evaluating the pharmacokinetics in GEM gliomas and normal brain tissue. Our goal was to develop a robust and reproducible method of kinetic analysis for the quantitative evaluation of tumor proliferation. METHODS: Dynamic (18)F-FLT PET imaging was performed for 60 min in glioma-bearing mice (n = 10) and in non-tumor-bearing control mice (n = 4) by use of a dedicated small-animal PET scanner. A 3-compartment, 4-parameter model was used to characterize (18)F-FLT kinetics in vivo. For compartmental analysis, the arterial input was measured by placing a region of interest over the left ventricular blood pool and was corrected for partial-volume averaging. The (18)F FLT "trapping" and tissue flux model parameters were correlated with measured uptake (percentage injected dose per gram [%ID/g]) values at 60 min. RESULTS: (18)F-FLT uptake values (%ID/g) at 1 h in brain tumors were significantly greater than those in control brains (mean +/- SD: 4.33 +/- 0.58 and 0.86 +/- 0.22, respectively; P < 0.0004). Kinetic analyses of the measured time-activity curves yielded independent, robust estimates of tracer transport and metabolism, with compartmental model-derived time-activity data closely fitting the measured data. Except for tracer transport, statistically significant differences were found between the applicable model parameters for tumors and normal brains. The tracer retention rate constant strongly correlated with measured (18)F-FLT uptake values (r = 0.85, P < 0.0025), whereas a more moderate correlation was found between net (18)F-FLT flux and (18)F-FLT uptake values (r = 0.61, P < 0.02). CONCLUSION: A clinically relevant mouse glioma model was characterized by both static and dynamic small-animal PET imaging of (18)F-FLT uptake. Time-activity curves were kinetically modeled to distinguish early transport from a subsequent tracer retention phase. Estimated (18)F-FLT rate constants correlated positively with %ID/g measurements. Dynamic evaluation of (18)F-FLT uptake offers a promising approach for noninvasively assessing cellular proliferation in vivo and for quantitatively monitoring new antiproliferation therapies. PMID- 18287266 TI - Quantitative measurements of bone remodeling using 99mTc-methylene diphosphonate bone scans and blood sampling. AB - Quantitative studies of bone using (99m)Tc-methylene diphosphonate ((99m)Tc-MDP) have a potentially valuable role in investigating the treatment of patients with metabolic bone disease. In this study we compared 3 different methods of measuring whole-skeleton (99m)Tc-MDP plasma clearance (K(bone)) in 12 osteoporotic postmenopausal women (mean age, 67.3 y) before participation in a clinical trial of an osteoporosis therapy. The aim was to compare the consistency and accuracy of the 3 methods before their use in evaluating the subjects' response to treatment. METHODS: Subjects were injected with 600 MBq (99m)Tc-MDP and 3 MBq (51)Cr-ethylenediaminetetraacetic acid ((51)Cr-EDTA) and whole-body bone scan images were acquired at 10 min, 1, 2, 3, and 4 h. Two-minute static images of the thighs were acquired immediately after the 1- to 4-h whole-body scans. Six blood samples were taken between 5 min and 4 h, and free (99m)Tc-MDP was measured using ultrafiltration. The glomerular filtration rate (GFR) was estimated from the (51)Cr-EDTA plasma curve. The methods used to evaluate K(bone) were (a) the area-under-the-curve (AUC) method, in which the GFR measurement was subtracted from the total (bone plus renal) clearance (K(total)) measured from the free (99m)Tc-MDP plasma curve; (b) the modified Brenner method, in which (99m)Tc-MDP renal clearance estimated from the whole-body counts was subtracted from the total clearance measured from the rate of elimination of tracer from soft tissue; and (c) the Patlak plot method, which was also used to derive regional values of K(bone) for the skull, spine, pelvis, arms, and legs. RESULTS: There was good agreement between the 3 methods of measuring K(bone). (mean K(bone) +/- SD: AUC method, 30.3 +/- 6.4 mL x min(-1); Brenner method, 31.1 +/- 5.8 mL x min(-1); Patlak method, 35.7 +/- 5.8 mL x min(-1)). The correlation coefficients between the methods varied from r = 0.767 (P = 0.004) to r = 0.805 (P = 0.002). Regional measurements of (99m)Tc-MDP clearance gave the following percentages of the whole-skeleton clearance: skull, 13.3%; spine, 16.6%; pelvis, 17.2%; arms, 11.1%; legs, 23.7%. CONCLUSION: The 3 methods gave consistent and accurate measurements of K(bone). The Patlak method can be used to study regional as well as total-skeleton values of K(bone). PMID- 18287267 TI - Adjuvant intraarterial injection of 131I-labeled lipiodol after resection of hepatocellular carcinoma: progress report of a case-control study with a 5-year minimal follow-up. AB - Recurrences after resection of hepatocellular carcinoma are frequent. A single postoperative injection of (131)I-labeled lipiodol in the hepatic artery was shown in 1999 by Lau and colleagues to be an effective adjuvant treatment, and those results were strengthened by our experience with a case-control study, reported in 2003. The goal of this paper is to update the 2003 results for a minimal follow-up of 5 y. METHODS: Between January 1999 and September 2001, 38 patients were given an adjuvant postoperative intraarterial injection of (131)I lipiodol and were matched (for Okuda group and tumor size) with 38 patients who had undergone resection between January 1997 and January 1999 without postoperative treatment. The 2 groups were similar. RESULTS: There were 28 recurrences in the control group and 22 in the (131)I-lipiodol group (not statistically significant), and the mean time of recurrence was 21 and 26.5 mo, respectively, after surgery (statistically significant). The number of recurrences was lower in the first 2 y in the (131)I-lipiodol group (statistically significant). Disease-free survival was better (P < 0.03) in the (131)I-lipiodol group than in the control group (2-, 3-, and 5-y rates [+/-95% confidence interval] of 77% +/- 7%, 63% +/- 8%, and 42% +/- 8.5%, respectively, for the (131)I-lipiodol group vs. 47% +/- 8%, 34% +/- 8%, and 27% +/- 8%, respectively, for the control group). Overall survival did not differ between the 2 groups (P = 0.09), even though there was a trend toward better survival in the (131)I-lipiodol group (2-, 3-, and 5-y rates of 76% +/- 7%, 68% +/- 7.5%, and 51% +/- 9%, respectively, vs. 68% +/- 7.5%, 53% +/- 8%, and 39% +/- 8%, respectively, in the control group). CONCLUSION: With a longer follow-up, the results of this retrospective case-control study still favor a single postoperative injection of (131)I-lipiodol. These retrospective findings point out the need for a large scale, prospective, randomized study. PMID- 18287268 TI - Quantitative imaging of estrogen receptor expression in breast cancer with PET and 18F-fluoroestradiol. AB - The PET compound (18)F-fluoroestradiol ((18)F-FES) has been developed and tested as an agent for the imaging of estrogen receptor (ER) expression in vivo. (18)F FES uptake has been shown to correlate with ER expression assayed in vitro by radioligand binding; however, immunohistochemistry (IHC) rather than radioligand binding is used most often to measure ER expression in clinical practice. We therefore compared (18)F-FES uptake with ER expression assayed in vitro by IHC with both qualitative and semiquantitative measures. METHODS: Seventeen patients with primary or metastatic breast cancer were studied with dynamic (18)F-FES PET; cancer tissue samples, collected close to the time of imaging, were assayed for ER expression by IHC. For each tumor, partial-volume-corrected measures of (18)F FES uptake were compared with ER expression measured by 3 different ER scoring methods: qualitative scoring (0-3+), the Allred score (0-10), and a computerized IHC index. RESULTS: There was excellent agreement (r = 0.99) between observers using IHC as well as the different methods of measuring ER content (P < 0.001). ER-negative tumors had (18)F-FES partial-volume-corrected standardized uptake values of less than 1.0, whereas ER-positive tumors had values above 1.1. Correlation coefficients for the different measures of ER content and the different measures of (18)F-FES uptake ranged from 0.57 to 0.73, with the best correlation being between the computerized IHC index and (18)F-FES partial-volume corrected standardized uptake values. CONCLUSION: Our results showed good agreement between (18)F-FES PET and ER expression measured by IHC. (18)F-FES imaging may be a useful tool for aiding in the assessment of ER status, especially in patients with multiple tumors or for tumors that are difficult to biopsy. PMID- 18287269 TI - Benefit of time-of-flight in PET: experimental and clinical results. AB - Significant improvements have made it possible to add the technology of time-of flight (TOF) to improve PET, particularly for oncology applications. The goals of this work were to investigate the benefits of TOF in experimental phantoms and to determine how these benefits translate into improved performance for patient imaging. METHODS: In this study we used a fully 3-dimensional scanner with the scintillator lutetium-yttrium oxyorthosilicate and a system timing resolution of approximately 600 ps. The data are acquired in list-mode and reconstructed with a maximum-likelihood expectation maximization algorithm; the system model includes the TOF kernel and corrections for attenuation, detector normalization, randoms, and scatter. The scatter correction is an extension of the model-based single scatter simulation to include the time domain. Phantom measurements to study the benefit of TOF include 27-cm- and 35-cm-diameter distributions with spheres ranging in size from 10 to 37 mm. To assess the benefit of TOF PET for clinical imaging, patient studies are quantitatively analyzed. RESULTS: The lesion phantom studies demonstrate the improved contrast of the smallest spheres with TOF compared with non-TOF and also confirm the faster convergence of contrast with TOF. These gains are evident from visual inspection of the images as well as a quantitative evaluation of contrast recovery of the spheres and noise in the background. The gains with TOF are higher for larger objects. These results correlate with patient studies in which lesions are seen more clearly and with higher uptake at comparable noise for TOF than with non-TOF. CONCLUSION: TOF leads to a better contrast-versus-noise trade-off than non-TOF but one that is difficult to quantify in terms of a simple sensitivity gain improvement: A single gain factor for TOF improvement does not include the increased rate of convergence with TOF nor does it consider that TOF may converge to a different contrast than non-TOF. The experimental phantom results agree with those of prior simulations and help explain the improved image quality with TOF for patient oncology studies. PMID- 18287271 TI - MRI-based correction for partial-volume effect improves detectability of intractable epileptogenic foci on 123I-iomazenil brain SPECT images. AB - (123)I-Iomazenil brain SPECT has been used for the detection of epileptogenic foci, especially when surgical intervention is considered. Although epileptogenic foci exhibit a decrease in (123)I-iomazenil accumulation, normal cerebral cortices often exhibit similar findings because of thin cortical ribbons, gray matter atrophy, or pathologic brain structures. In the present study, we created (123)I-iomazenil SPECT images corrected for gray matter volume using MRI and tested whether the detectability of the epileptogenic foci improved. METHODS: Seven patients (1 male patient and 6 female patients; mean age +/- SD, 34 +/- 17 y) with intractable epilepsy were surgically treated by resecting the cerebral cortex after surface electroencephalography. Histopathologic examination of the resected specimens and a good outcome after surgery indicated that the resected lesions were epileptogenic foci. These patients underwent (123)I-iomazenil SPECT and 3-dimensional T1-weighted MRI examinations before their operations. Each SPECT image was coregistered to the corresponding MR image, and its partial volume effect (PVE) was corrected on a voxel-by-voxel basis with a smoothed gray matter distribution image. Four nuclear medicine physicians visually evaluated the (123)I-iomazenil SPECT images with and without the PVE correction. The SPECT count ratio of the suspected focus to the contralateral cerebral cortex was evaluated as an asymmetry index (%) based on the volume of interest. RESULTS: The sensitivity, specificity, and accuracy of focus detection by visual assessment were higher after PVE correction (88%, 99%, and 98%, respectively) than before correction (50%, 92%, and 87%, respectively). The mean asymmetry index for the surgically resected lesions was significantly higher on the PVE-corrected SPECT images (22%) than on the PVE-uncorrected ones (16%) (P = 0.006). CONCLUSION: MRI based PVE correction for (123)I-iomazenil brain SPECT improves the sensitivity and specificity of the detection of cortical epileptogenic foci in patients with intractable epilepsy. PMID- 18287270 TI - Multicenter standardized 18F-FDG PET diagnosis of mild cognitive impairment, Alzheimer's disease, and other dementias. AB - This multicenter study examined (18)F-FDG PET measures in the differential diagnosis of Alzheimer's disease (AD), frontotemporal dementia (FTD), and dementia with Lewy bodies (DLB) from normal aging and from each other and the relation of disease-specific patterns to mild cognitive impairment (MCI). METHODS: We examined the (18)F-FDG PET scans of 548 subjects, including 110 healthy elderly individuals ("normals" or NLs), 114 MCI, 199 AD, 98 FTD, and 27 DLB patients, collected at 7 participating centers. Individual PET scans were Z scored using automated voxel-based comparison with generation of disease-specific patterns of cortical and hippocampal (18)F-FDG uptake that were then applied to characterize MCI. RESULTS: Standardized disease-specific PET patterns were developed that correctly classified 95% AD, 92% DLB, 94% FTD, and 94% NL. MCI patients showed primarily posterior cingulate cortex and hippocampal hypometabolism (81%), whereas neocortical abnormalities varied according to neuropsychological profiles. An AD PET pattern was observed in 79% MCI with deficits in multiple cognitive domains and 31% amnesic MCI. (18)F-FDG PET heterogeneity in MCI with nonmemory deficits ranged from absent hypometabolism to FTD and DLB PET patterns. CONCLUSION: Standardized automated analysis of (18)F FDG PET scans may provide an objective and sensitive support to the clinical diagnosis in early dementia. PMID- 18287272 TI - Virtual-pinhole PET. AB - We proposed and tested a novel geometry for PET system design analogous to pinhole SPECT called the virtual-pinhole PET (VP-PET) geometry to determine whether it could provide high-resolution images. METHODS: We analyzed the effects of photon acolinearity and detector sizes on system resolution and extended the empiric formula for reconstructed image resolution of conventional PET proposed earlier to predict the resolutions of VP-PET. To measure the system resolution of VP-PET, we recorded coincidence events as a (22)Na point source was stepped across the coincidence line of response between 2 detectors made from identical arrays of 12 x 12 lutetium oxyorthosilicate crystals (each measuring 1.51 x 1.51 x 10 mm(3)) separated by 565 mm. To measure reconstructed image resolution, we built 4 VP-PET systems using 4 types of detectors (width, 1.51-6.4 mm) and imaged 4 point sources of (64)Cu (half-life = 12.7 h to allow a long acquisition time). Tangential and radial resolutions were measured and averaged for each source and each system. We then imaged a polystyrene plastic phantom representing a 2.5-cm thick cross-section of isolated breast volume. The phantom was filled with an aqueous solution of (64)Cu (713 kBq/mL) in which the following were imbedded: 4 spheric tumors ranging from 1.8 to 12.6 mm in inner diameter (ID), 6 micropipettes (0.7- or 1.1-mm ID filled with (64)Cu at 5x, 20x, or 50x background), and a 10.0-mm outer-diameter cold lesion. RESULTS: The shape and measured full width at half maximum of the line spread functions agree well with the predicted values. Measured reconstructed image resolution (2.40-3.24 mm) was +/-6% of the predicted value for 3 of the 4 systems. In one case, the difference was 12.6%, possibly due to underestimation of the block effect from the low resolution detector. In phantom experiments, all spheric tumors were detected. Small line sources were detected if the activity concentration is at least 20x background. CONCLUSION: We have developed and characterized a novel geometry for PET. A PET system following the VP-PET geometry provides high-resolution images for objects near the system's high-resolution detectors. This geometry may lead to the development of special-purpose PET systems or resolution-enhancing insert devices for conventional PET scanners. PMID- 18287273 TI - Recommendations on the use of 18F-FDG PET in oncology. AB - The rationale was to develop recommendations on the use of (18)F-FDG PET in breast, colorectal, esophageal, head and neck, lung, pancreatic, and thyroid cancer; lymphoma, melanoma, and sarcoma; and unknown primary tumor. Outcomes of interest included the use of (18)F-FDG PET for diagnosing, staging, and detecting the recurrence or progression of cancer. METHODS: A search was performed to identify all published randomized controlled trials and systematic reviews in the literature. An additional search was performed to identify relevant unpublished systematic reviews. These publications comprised both retrospective and prospective studies of varied methodologic quality. The anticipated consequences of false-positive and false-negative tests when evaluating clinical usefulness, and the impact of (18)F-FDG PET on the management of cancer patients, were also reviewed. RESULTS AND CONCLUSION: (18)F-FDG PET should be used as an imaging tool additional to conventional radiologic methods such as CT or MRI; any positive finding that could lead to a clinically significant change in patient management should be confirmed by subsequent histopathologic examination because of the risk of false-positive results. (18)F-FDG PET should be used in the appropriate clinical setting for the diagnosis of head and neck, lung, or pancreatic cancer and for unknown primary tumor. PET is also indicated for staging of breast, colon, esophageal, head and neck, and lung cancer and of lymphoma and melanoma. In addition, (18)F-FDG PET should be used to detect recurrence of breast, colorectal, head and neck, or thyroid cancer and of lymphoma. PMID- 18287274 TI - 18F-labeled BBN-RGD heterodimer for prostate cancer imaging. AB - Both bombesin (BBN) analogs and cyclic RGD peptides have been suitably radiolabeled for prostate cancer imaging. However, the limited expression of gastrin-releasing peptide receptor (GRPR) and integrin alpha(v)beta(3) as well as unfavorable in vivo kinetics limited further applications of these imaging agents. We hypothesize that a peptide ligand recognizing both GRPR and integrin will be advantageous because of its dual-receptor-targeting ability. METHODS: A BBN-RGD heterodimer was synthesized from bombesin(7-14) and c(RGDyK) through a glutamate linker and then labeled with (18)F via the N-succinimidyl-4-(18)F fluorobenzoate ((18)F-SFB) prosthetic group. The receptor-binding characteristics and tumor-targeting efficacy of (18)F-FB-BBN-RGD were tested in vitro and in vivo. RESULTS: FB-BBN-RGD had comparable integrin alpha(v)beta(3)-binding affinity with c(RGDyK) and comparable GRPR-binding affinity with BBN(7-14). (18)F FB-BBN-RGD had significantly higher tumor uptake compared with monomeric RGD and monomeric BBN peptide tracer analogs at all time points examined. The PC-3 tumor uptake of (18)F-FB-BBN-RGD was inhibited only partially in the presence of an excess amount of unlabeled BBN(7-14) or c(RGDyK) but was blocked completely in the presence of both BBN(7-14) and c(RGDyK). Compared with (18)F-FB-BBN and (18)F FB-RGD, (18)F-FB-BBN-RGD also had improved pharmacokinetics, resulting in a significantly higher imaging quality. CONCLUSION: Dual integrin alpha(v)beta(3) and GRPR recognition showed significantly improved tumor-targeting efficacy and pharmacokinetics compared with (18)F-labeled RGD and BBN analogs. The same heterodimeric ligand design may also be applicable to other receptor system combinations and other imaging modalities. PMID- 18287275 TI - Whole-body biodistribution and radiation dosimetry of the human cannabinoid type 1 receptor ligand 18F-MK-9470 in healthy subjects. AB - The cannabinoid type-1 (CB1) receptor is one of the most abundant G-coupled protein receptors in the human body and is responsible for signal transduction of both endogenous and exogenous cannabinoids. The endocannabinoid system is strongly implicated in regulation of homeostasis and several neuropsychiatric disorders, obesity, and associated comorbidities, such as dyslipidemia and metabolic syndrome. We have used whole-body PET/CT to characterize the biodistribution and dosimetry of a novel high-affinity, subtype-selective radioligand, (18)F-MK-9470, in healthy male and female subjects. METHODS: Eight nonobese subjects (5 men, 3 women; age, 22-54 y) underwent serial whole-body PET/CT for 6 h after a bolus injection of 251 +/- 25 MBq (18)F-MK-9470 (N-[2-(3 cyano-phenyl)-3-(4-(2-(18)F-fluorethoxy)phenyl)-1-methylpropyl]-2-(5-methyl-2 pyridyloxy)-2-methylproponamide). Source organs were delineated 3-dimensionally using the combined morphologic and functional data. Residence times were derived from time-activity profiles using both the trapezoid rule and curve fitting. Individual organ doses and effective doses were determined using the OLINDA software package, with different approaches for gastrointestinal and urinary excretion modeling. RESULTS: (18)F-MK-9470 is taken up slowly in the brain, reaching a plateau at approximately 90-120 min after bolus injection and is excreted predominantly through the hepatobiliary system. The gallbladder, upper large intestine, small intestine, and liver are the organs with the highest absorbed dose (average: 159, 98, 87, and 86 microGy/MBq, respectively). The mean effective dose (ED) was 22.8 +/- 4.3 microSv/MBq, indicating relatively low intersubject variability and a mean value in the range of many commercially available (18)F-labeled radiopharmaceuticals. Brain uptake was relatively high compared with that of existing central nervous system ligands for other receptors, between 3.2% and 4.9% of the injected dose. CONCLUSION: The estimated radiation burden of (18)F-MK-9470 for PET CB1 receptor imaging shows relatively low variability between subjects and has an acceptable ED, which allows multiple serial cerebral scans of good image quality, while remaining within the risk category class II-b defined by the World Health Organization and the International Commission for Radiation Protection for a standard injected activity (185-370 MBq). PMID- 18287276 TI - An intramolecular FRET system monitors fingers subdomain opening in Klentaq1. AB - A major goal of polymerase research is to determine the mechanism through which a nucleotide complementary to a templating DNA base is selected and delivered to the polymerase active site. Structural evidence suggests a large open-to-closed conformational change affecting the fingers subdomain as being crucial to the process. We previously designed a FRET system capable of measuring the rate of fingers subdomain closure in the presence of correct nucleotide. However, this FRET system was limited in that it could not directly measure the rate of fingers subdomain opening by FRET after polymerization or in the absence of DNA. Here we report the development of a new system capable of measuring both fingers subdomain closure and reopening by FRET, and show that the rate of fingers subdomain opening is limited only by the rate of polymerization. We anticipate that this system will scale down to the single molecule level, allowing measurement of fingers subdomain movements in the presence of incorrect nucleotide and in the absence of DNA. PMID- 18287277 TI - Rational design of a novel calcium-binding site adjacent to the ligand-binding site on CD2 increases its CD48 affinity. AB - Electrostatic interactions are important for molecular recognition processes including Ca2+-binding and cell adhesion. To understand these processes, we have successfully introduced a novel Ca2+-binding site into the non-Ca2+-dependent cell adhesion protein CD2 using our criteria that are specifically tailored to the structural and functional properties of the protein environment and charged adhesion surface. This designed site with ligand residues exclusively from the beta-sheets selectively binds to Ca2+ and Ln3+ over other mono- and divalent cations. While Ca2+ and Ln3+ binding specifically alters the local environment of the designed Ca2+-binding site, the designed protein undergoes a significantly smaller conformation change compared with those observed in naturally occurring Ca2+-binding sites that are composed of at least part of the flexible loop and helical regions. In addition, the CD2-CD48-binding affinity increased approximately threefold after protein engineering, suggesting that the cell adhesion of CD2 can be modulated by altering the local electrostatic environment. The study provides site-specific information for regulating cell adhesion within CD2 and gives insight into the structural factors required for Ca2+-modulated biological processes. PMID- 18287278 TI - Crystal structure of LpxC from Pseudomonas aeruginosa complexed with the potent BB-78485 inhibitor. AB - The cell wall in Gram-negative bacteria is surrounded by an outer membrane comprised of charged lipopolysaccharide (LPS) molecules that prevent entry of hydrophobic agents into the cell and protect the bacterium from many antibiotics. The hydrophobic anchor of LPS is lipid A, the biosynthesis of which is essential for bacterial growth and viability. UDP-3-O-(R-3-hydroxymyristoyl)-N acetylglucosamine deacetylase (LpxC) is an essential zinc-dependant enzyme that catalyzes the conversion of UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine to UDP-3-O-(R-3-hydroxymyristoyl)glucosamine and acetate in the biosynthesis of lipid A, and for this reason, LpxC is an attractive target for antibacterial drug discovery. Here we disclose a 1.9 A resolution crystal structure of LpxC from Pseudomonas aeruginosa (paLpxC) in a complex with the potent BB-78485 inhibitor. To our knowledge, this is the first crystal structure of LpxC with a small molecule inhibitor that shows antibacterial activity against a wide range of Gram negative pathogens. Accordingly, this structure can provide important information for lead optimization and rational design of the effective small-molecule LpxC inhibitors for successful treatment of Gram-negative infections. PMID- 18287279 TI - Engineering and characterization of a single chain surrogate light chain variable domain. AB - The surrogate light chain (SLC) is a key regulator of B cell development in the bone marrow, resulting in mature B cells that produce antibodies that are capable of interacting with antigens. The SLC comprises two noncovalently interacting proteins: VpreB and 14.1. We engineered a construct to represent the complete immunoglobulin-like domain of the SLC variable domain in a single protein chain that could be bacterially expressed. In this construct, the incomplete immunoglobulin domain of VpreB (residues 1-102) was linked to the J-segment of 14.1 (residues 40-53), which provided one beta-strand to complete the V-like domain (VpreBJ). Because VpreBJ has the interface to VH chains, but lacks the unique region of 14.1, which is important for SLC signaling, we predict that a properly folded VpreBJ would have the potential to act as a dominant negative mutant of the surrogate light chain. X-ray crystallography of VpreBJ at 2.0 A resolution showed that the engineering was successful. With its two beta-pleated sheets, packed face-to-face, the single chain VpreBJ resembles a mature light chain immunoglobulin V-domain (VL). The surface that would normally interact with the VH chain interacts with a crystallographically related VpreBJ molecule. The presence of dimeric species in solution was verified by analytical ultracentrifugation. VpreBJ is easily overexpressed in bacteria, while retaining the native conformation of an immunoglobulin domain, and thus may serve as an important reagent for future studies in B-cell development. PMID- 18287280 TI - Prediction of reversibly oxidized protein cysteine thiols using protein structure properties. AB - Protein cysteine thiols can be divided into four groups based on their reactivities: those that form permanent structural disulfide bonds, those that coordinate with metals, those that remain in the reduced state, and those that are susceptible to reversible oxidation. Physicochemical parameters of oxidation susceptible protein thiols were organized into a database named the Balanced Oxidation Susceptible Cysteine Thiol Database (BALOSCTdb). BALOSCTdb contains 161 cysteine thiols that undergo reversible oxidation and 161 cysteine thiols that are not susceptible to oxidation. Each cysteine was represented by a set of 12 parameters, one of which was a label (1/0) to indicate whether its thiol moiety is susceptible to oxidation. A computer program (the C4.5 decision tree classifier re-implemented as the J48 classifier) segregated cysteines into oxidation-susceptible and oxidation-non-susceptible classes. The classifier selected three parameters critical for prediction of thiol oxidation susceptibility: (1) distance to the nearest cysteine sulfur atom, (2) solvent accessibility, and (3) pKa. The classifier was optimized to correctly predict 136 of the 161 cysteine thiols susceptible to oxidation. Leave-one-out cross validation analysis showed that the percent of correctly classified cysteines was 80.1% and that 16.1% of the oxidation-susceptible cysteine thiols were incorrectly classified. The algorithm developed from these parameters, named the Cysteine Oxidation Prediction Algorithm (COPA), is presented here. COPA prediction of oxidation-susceptible sites can be utilized to locate protein cysteines susceptible to redox-mediated regulation and identify possible enzyme catalytic sites with reactive cysteine thiols. PMID- 18287281 TI - Unfolding the fold of cyclic cysteine-rich peptides. AB - We propose a method to extensively characterize the native state ensemble of cyclic cysteine-rich peptides. The method uses minimal information, namely, amino acid sequence and cyclization, as a topological feature that characterizes the native state. The method does not assume a specific disulfide bond pairing for cysteines and allows the possibility of unpaired cysteines. A detailed view of the conformational space relevant for the native state is obtained through a hierarchic multi-resolution exploration. A crucial feature of the exploration is a geometric approach that efficiently generates a large number of distinct cyclic conformations independently of one another. A spatial and energetic analysis of the generated conformations associates a free-energy landscape to the explored conformational space. Application to three long cyclic peptides of different folds shows that the conformational ensembles and cysteine arrangements associated with free energy minima are fully consistent with available experimental data. The results provide a detailed analysis of the native state features of cyclic peptides that can be further tested in experiment. PMID- 18287282 TI - Structural constraints on autoprocessing of the human nucleoporin Nup98. AB - Nucleoporin Nup98, a 98-kDa protein component of the nuclear pore complex, plays an important role in both protein and RNA transport. During its maturation process, Nup98 undergoes post-translational autoproteolysis, which is critical for targeting to the NPC. Here we present high-resolution crystal structures of the C-terminal autoproteolytic domains of Nup98 (2.3 A for the wild type and 1.9 A for the S864A precursor), and propose a detailed autoproteolysis mechanism through an N-O acyl shift. Structural constraints are found at the autocleavage site, and could thus provide a driving force for autocleavage at the scissile peptide bond. Such structural constraints appear to be generated, at least in part, by anchoring a conserved phenylalanine side chain into a highly conserved hydrophobic pocket at the catalytic site. Our high-resolution crystal structures also reveal that three highly conserved residues, Tyr866, Gly867, and Leu868, provide most of the interactions between the autoproteolytic domain and the C terminal tail. These results suggest that Nup98 may represent a new subtype of protein that utilizes autoprocessing to control biogenesis pathways and intracellular translocation. PMID- 18287283 TI - The structure of the lipid-embedded potassium channel voltage sensor determined by double-electron-electron resonance spectroscopy. AB - A four-pulse electron paramagnetic resonance experiment was used to measure long range inter-subunit distances in reconstituted KvAP, a voltage-dependent potassium (Kv) channel. The measurements have allowed us to reach the following five conclusions about the native structure of the voltage sensor of KvAP. First, the S1 helix of the voltage sensor engages in a helix packing interaction with the pore domain. Second, the crystallographically observed antiparallel helix turn-helix motif of the voltage-sensing paddle is retained in the membrane embedded voltage sensor. Third, the paddle is oriented in such a way as to expose one face to the pore domain and the opposite face to the membrane. Fourth, the paddle and the pore domain appear to be separated by a gap that is sufficiently wide for lipids to penetrate between the two domains. Fifth, the critical voltage sensing arginine residues on the paddle appear to be lipid exposed. These results demonstrate the importance of the membrane for the native structure of Kv channels, suggest that lipids are an integral part of their native structure, and place the voltage-sensing machinery into a complex lipid environment near the pore domain. PMID- 18287284 TI - Intersubunit linker length as a modifier of protein stability: crystal structures and thermostability of mutant TRAP. AB - The ability of proteins to self-assemble into complex, functional nanoscale structures is expected to become of significant use in the manufacture of artificial nanodevices with a wide range of novel applications. The bacterial protein TRAP has potential uses as a nanoscale component as it is ring-shaped, with a central, modifiable cavity. Furthermore, it can be engineered to make a ring of 12-fold symmetry, which is advantageous for packing into two-dimensional arrays. The 12mer form of TRAP is made by linking multiple subunits together on the same polypeptide, but the usefulness of the 12mers described to date is limited by their poor stability. Here we show that, by altering the length of the peptide linker between subunits, the thermostability can be significantly improved. Since the subunit interfaces of the different 12mers are essentially identical, stabilization arises from the reduction of strain in the linkers. Such a simple method of controlling the stability of modular proteins may have wide applications, and demonstrates the lack of absolute correlation between interactions observable by crystallography and the internal energy of a complex. PMID- 18287286 TI - Is lipopolysaccharide a factor in infectivity of Chlamydia trachomatis? AB - Lipopolysaccharide (LPS) is a major surface component of Chlamydia trachomatis, as with all Gram-negative bacteria. The effect of C. trachomatis LPS on C. trachomatis infectivity of human epithelial cells was investigated. C. trachomatis LPS and C. trachomatis LPS antibody significantly reduced infectivity, mostly in a dose-dependent manner. As the structure of LPS in C. trachomatis is simple and consists only of lipid A and 3-deoxy-D-manno octulosonic acid (Kdo), we investigated whether lipid A or Kdo was inhibitory to chlamydial infectivity. Polymyxin B, as a lipid A inhibitor, and Kdo considerably reduced C. trachomatis infectivity. With all the LPS inhibitors used, there was greater inhibition against serovar E than serovar LGV. These results suggest a role for LPS in chlamydial infectivity. Elucidation of how LPS acts in infectivity and identification of host-cell receptors would help in understanding pathogenicity. PMID- 18287285 TI - The solution structure of ZNF593 from Homo sapiens reveals a zinc finger in a predominantly unstructured protein. AB - Here, we report the solution structure of ZNF593, a protein identified in a functional study as a negative modulator of the DNA-binding activity of the Oct-2 transcription factor. ZNF593 contains a classic C(2)H(2) zinc finger domain flanked by about 40 disordered residues on each terminus. Although the protein contains a high degree of intrinsic disorder, the structure of the zinc finger domain was resolved by NMR spectroscopy without a need for N- or C-terminal truncations. The tertiary structure of the zinc finger domain is composed of a beta-hairpin that positions the cysteine side chains for zinc coordination, followed by an atypical kinked alpha-helix containing the two histidine side chain ligands. The structural topology of ZNF593 is similar to a fragment of the double-stranded RNA-binding protein Zfa and the C-terminal zinc finger of MBP-1, a human enhancer binding protein. The structure presented here will provide a guide for future functional studies of how ZNF593 negatively modulates the DNA binding activity of Oct-2, a POU domain-containing transcription factor. Our work illustrates the unique capacity of NMR spectroscopy for structural analysis of folded domains in a predominantly disordered protein. PMID- 18287287 TI - Differences in virulence attributes between cytolethal distending toxin positive and negative Campylobacter jejuni strains. AB - Campylobacter jejuni is a common gastrointestinal bacterial pathogen. Although cytolethal distending toxin (CDT) is proposed to be an important virulence determinant of this pathogen, how CDT(+) and CDT(-) strains differ in their biological properties remains largely unknown. The virulence properties of CDT(+) and CDT(-) strains were studied on HeLa cells and in the suckling mouse model. Presence of the cdtB gene in Campylobacter species was determined by PCR. Five each of CDT(+) and CDT(-) C. jejuni strains were subjected to adherence, invasion and cytotoxicity assay on the HeLa cell line. Bacterial culture supernatants with and without CDT activity were inoculated intragastrically into 2-day-old suckling mice. The mice were sacrificed within 48 h. Histopathological examination of stomach, jejunum, ileum and colon was performed by haematoxylin/eosin staining. cdtB was detected in 88 % and 14 % of C. jejuni and Campylobacter coli strains, respectively. CDT(+) C. jejuni strains adhered to and invaded HeLa cells in significantly higher numbers than CDT(-) strains [CDT(+) vs CDT(-), adherence 2.7 x 10(4)+/-3.5 x 10(4) vs 2.7 x 10(2)+/-1.9 x 10(2); invasion 1.0 x 10(3)+/-1.3 x 10(3) vs 1.4 x 10(1)+/-3.1 x 10(1); P<0.01]. Culture supernatants of all CDT(+) strains demonstrated CDT activity on HeLa cells. Mice inoculated with supernatant containing CDT activity had moderate to severe pathology in different parts of their gastrointestinal tract, with the colon being the major target. Mice inoculated with supernatant lacking CDT activity showed no significant pathology in the gastrointestinal tract. The results demonstrate that CDT(+) C. jejuni strains adhere to and invade epithelial cells more efficiently than CDT(-) strains. CDT is responsible for intestinal pathology and the colon is the major target. PMID- 18287288 TI - Recognition of pneumococcal isolates by antisera raised against PspA fragments from different clades. AB - Pneumococcal surface protein A (PspA) is an important vaccine candidate against pneumococcal infections, capable of inducing protection in different animal models. Based on its structural diversity, it has been suggested that a PspA based vaccine should contain at least one fragment from each of the two major families (family 1, comprising clades 1 and 2, and family 2, comprising clades 3, 4 and 5) in order to elicit broad protection. This study analysed the recognition of a panel of 35 pneumococcal isolates bearing different PspAs by antisera raised against the N-terminal regions of PspA clades 1 to 5. The antiserum to PspA clade 4 was found to show the broadest cross-reactivity, being able to recognize pneumococcal strains containing PspAs of all clades in both families. The cross reactivity of antibodies elicited against a PspA hybrid including the N-terminal region of clade 1 fused to a shorter and more divergent fragment (clade-defining region, or CDR) of clade 4 (PspA1-4) was also tested, and revealed a strong recognition of isolates containing clades 1, 4 and 5, and weaker reactions with clades 2 and 3. The analysis of serum reactivity against different PspA regions further revealed that the complete N-terminal region rather than just the CDR should be included in an anti-pneumococcal vaccine. A PspA-based vaccine is thus proposed to be composed of the whole N-terminal region of clades 1 and 4, which could also be expressed as a hybrid protein. PMID- 18287289 TI - Glycosyltransferase: a specific marker for the discrimination of Bacillus anthracis from the Bacillus cereus group. AB - Bacillus anthracis, the aetiological agent of anthrax, has been taxonomically classified with the Bacillus cereus group, which comprises B. cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus pseudomycoides and Bacillus weihenstephanensis. Although the pathogenesis and ecological manifestations may be different, B. anthracis shares a high degree of DNA sequence similarity with its group member species. As a result, the discrimination of B. anthracis from its close relatives in the B. cereus group is still quite difficult. Suppression subtractive hybridization (SSH) was performed to search for genomic differences between a B. anthracis Korean isolate CR and the most closely related B. cereus type strain KCTC 3624(T). Two-hundred and five B. anthracis CR clones obtained by SSH underwent Southern hybridization, and comparative sequences were analysed using the blast program from the National Center for Biotechnology Information (NCBI). Subsequently, primer sets based on the glycosyltransferase group 1 family protein gene specific to B. anthracis were designed from the sequences of subtracted clones, and their specificities were evaluated using eight B. anthracis, 33 B. cereus, 10 B. thuringiensis, six B. mycoides, one B. pseudomycoides, one B. weihenstephanensis and 19 strains from 11 other representative Bacillus species. PCR primers specific for the glycosyltransferase group 1 family protein gene did not amplify the desired products from any of the Bacillus strains under examination, except B. anthracis alone. These findings may be useful in the future development of efficient diagnostic tools for the rapid identification of B. anthracis from other members of the B. cereus group. PMID- 18287290 TI - Assessment of the clinical utility of serial beta-D-glucan concentrations in patients with persistent neutropenic fever. AB - The performance of the Fungitell assay was investigated in 100 patients with haematological malignancy undergoing chemotherapy who developed antibiotic unresponsive neutropenic fever (AUNF). Serum beta-D-glucan (BG) concentrations were significantly elevated on the first day of AUNF and all subsequent alternate days to day 10 in 38 patients who developed an invasive fungal infection (IFI) compared to 42 patients remaining free of such infections. The mean and median values of BG were 171.9+/-29.6 and 95.8 pg ml(-1), respectively, for patients with IFI and 64.4+/-17.1 and 32.9 pg ml(-1) for patients with only AUNF (P<0.0001). The differences remained significant over the 10 days despite antifungal therapy. The occurrence of > or =2 sequential concentrations of > or =80 pg ml(-1) ('positive' test) was found to give the best overall option for diagnosis, with an accuracy of 81.3%, sensitivity of 86.8%, positive predictive value of 76.7% and negative predictive value of 86.5%. Of the patients with an IFI, 78% developed a positive test at or before the clinical diagnosis was made - this occurred at a mean (range) of 1.25 (-14 to +14) days prior to the IFI diagnosis. By starting sampling of blood from the first day of neutropenia rather than from the first day of AUNF, 50% of the patients with subsequent IFI would have been identified 5 days earlier. Increasing sampling to daily from alternate day frequency did not further improve this earlier timing of an IFI diagnosis. A greater proportion of patients with persistent high levels of BG without overt IFI had severe enterocyte damage or mucositis than those with lower levels of BG without IFI (P=0.002). If the results of the initial BG test had been acted on to change antifungal therapy, discontinuation would have been inappropriate in 30% of patients and would have delayed definitive antifungal therapy. Although the findings for the cohort of patients studied are very useful, there is inter patient variability in the test's performance. An holistic diagnostic approach is therefore necessary to interpret the test results optimally. Future studies should address this in further detail as well as the impact of empirical antifungal drug use and patient outcome. PMID- 18287291 TI - Improving molecular detection of Candida DNA in whole blood: comparison of seven fungal DNA extraction protocols using real-time PCR. AB - The limitations of classical diagnostic methods for invasive Candida infections have led to the development of molecular techniques such as real-time PCR to improve diagnosis. However, the detection of low titres of Candida DNA in blood from patients with candidaemia requires the use of extraction methods that efficiently lyse yeast cells and recover small amounts of DNA suitable for amplification. In this study, a Candida-specific real-time PCR assay was used to detect Candida albicans DNA in inoculated whole blood specimens extracted using seven different extraction protocols. The yield and quality of total nucleic acids were estimated using UV absorbance, and specific recovery of C. albicans genomic DNA was estimated quantitatively in comparison with a reference (Qiagen kit/lyticase) method currently in use in our laboratory. The extraction protocols were also compared with respect to sensitivity, cost and time required for completion. The TaqMan PCR assay used to amplify the DNA extracts achieved high levels of specificity, sensitivity and reproducibility. Of the seven extraction protocols evaluated, only the MasterPure yeast DNA extraction reagent kit gave significantly higher total nucleic acid yields than the reference method, although nucleic acid purity was highest using either the reference or YeaStar genomic DNA kit methods. More importantly, the YeaStar method enabled C. albicans DNA to be detected with highest sensitivity over the entire range of copy numbers evaluated, and appears to be an optimal method for extracting Candida DNA from whole blood. PMID- 18287292 TI - A comparative study of three different PCR assays for detection of Mycoplasma genitalium in urogenital specimens from men and women. AB - The aim of this study was to compare conventional 16S rRNA gene PCR, real-time 16S rRNA gene PCR and real-time Mycoplasma genitalium adhesin protein (MgPa) gene PCR as detection methods for M. genitalium infection. The study also determined the prevalence of M. genitalium in male and female patients attending a sexually transmitted infections clinic in a rural area in the west of Sweden. First void urine (FVU) and/or urethral swabs were collected from 381 men, and FVU and/or cervical swabs and/or urethral swabs were collected from 298 women. A total of 213 specimens were used in the PCR comparative study: 98 consecutively sampled specimens from patients enrolled in the prevalence study, 36 consecutively sampled specimens from patients with symptoms of urethritis and 79 specimens from patients positive for M. genitalium by real-time MgPa gene PCR in the prevalence study. A true-positive M. genitalium DNA specimen was defined as either a specimen positive in any two PCR assays or a specimen whose PCR product was verified by DNA sequencing. The prevalence of M. genitalium infection in men and women was 27/381 (7.1 %) and 23/298 (7.7 %), respectively. In the PCR comparative study, M. genitalium DNA was detected in 61/76 (80.3 %) of true-positive specimens by conventional 16S rRNA gene PCR, in 52/76 (68.4 %) by real-time 16S rRNA gene PCR and in 74/76 (97.4 %) by real-time MgPa gene PCR. Real-time MgPa gene PCR thus had higher sensitivity compared with conventional 16S rRNA gene PCR and had considerably increased sensitivity compared with real-time 16S rRNA gene PCR for detection of M. genitalium DNA. Real-time MgPa gene PCR is well suited for the clinical diagnosis of M. genitalium. PMID- 18287293 TI - Performance of chromID ESBL, a chromogenic medium for detection of Enterobacteriaceae producing extended-spectrum beta-lactamases. AB - The chromogenic agar medium chromID ESBL (bioMerieux) was compared with BLSE agar medium (AES) for selective isolation and presumptive identification of extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae from clinical samples. A total of 765 samples (468 rectal swabs, 255 urine samples and 42 pulmonary aspirations) obtained from 547 patients was processed. All bacterial strains isolated on either medium were further characterized using biochemical tests, and ESBL producers were confirmed by synergy testing. Genetic characterization of ESBL genes was determined by PCR. A total of 33 ESBL producing Enterobacteriaceae strains [Escherichia coli (n=16), Klebsiella pneumoniae (n=8), Enterobacter spp. (n=3), Citrobacter spp. (n=5) and Proteus mirabilis (n=1)] was recovered. The sensitivity after 24 h incubation was 88 % for chromID ESBL and 85 % for BLSE agar. At 48 h, the sensitivity of chromID ESBL increased to 94 % and was higher than that obtained with BLSE agar. The positive predictive value at 24 h for chromID ESBL was 38.7 % [95 % confidence interval (95 % CI) 28.3 -50.2 %)], which was significantly higher than that for BLSE agar [15.4 %, 95 % CI 10.1 -21.5 %]. On both media, false-positive results were mostly due to Pseudomonas aeruginosa and to Enterobacteriaceae overproducing chromosomal cephalosporinase (Enterobacter spp.) or a chromosomal penicillinase (Klebsiella oxytoca). This study showed that chromID ESBL, a ready-to-use chromogenic selective medium, is sensitive and specific for rapid, presumptive identification of ESBL-producing Enterobacteriaceae. Its chromogenic properties and its selectivity are particularly useful in specimens containing resident associated flora. PMID- 18287294 TI - The TUBEX test detects not only typhoid-specific antibodies but also soluble antigens and whole bacteria. AB - TUBEX (IDL Biotech) is a 5 min semiquantitative colorimetric test for typhoid fever, a widely endemic disease. TUBEX detects anti-Salmonella O9 antibodies from a patient's serum by the ability of these antibodies to inhibit the binding between an indicator antibody-bound particle and a magnetic antigen-bound particle. Herein, we report that TUBEX could also be used to specifically detect soluble O9 lipopolysaccharide in antigen-spiked buffer by the ability of the antigen to inhibit the same binding between the particles. Sensitivity of antigen detection was improved (8-31 mug ml(-1)) by using a modified protocol in which the test sample was mixed with the indicator particles first, rather than with the magnetic particles as for antibody detection. The antigen was also detectable in spiked serum and urine samples, albeit less well (2-4-fold) than in buffer generally. However, no antigen was detected from six typhoid sera examined, all of which had anti-O9 antibodies. In addition, whole organisms of Salmonella Typhi (15 strains) and Salmonella Enteritidis (6 strains) (both O9(+) Salmonella), grown in simulated blood broths or on MacConkey agar, were also detectable by TUBEX when suspended at >9 x 10(8) organisms ml(-1). Expectedly, Salmonella Paratyphi A (7 strains), Salmonella Typhimurium (1 strain) and Escherichia coli (2 strains) were negative in the test. Thus, the same TUBEX kit may be used in several ways both serologically and microbiologically for the rapid diagnosis of typhoid fever. However, validation of the newer applications will require the systematic examination of real patient and laboratory materials. PMID- 18287295 TI - A quadruplex real-time PCR assay for the detection of Yersinia pestis and its plasmids. AB - Yersinia pestis, the aetiological agent of the plague, causes sporadic disease in endemic areas of the world and is classified as a National Institute of Allergy and Infectious Diseases Category A Priority Pathogen because of its potential to be used as a bioweapon. Health departments, hospitals and government agencies need the ability to rapidly identify and characterize cultured isolates of this bacterium. Assays have been developed to perform this function; however, they are limited in their ability to distinguish Y. pestis from Yersinia pseudotuberculosis. This report describes the creation of a real-time PCR assay using Taqman probes that exclusively identifies Y. pestis using a unique target sequence of the yihN gene on the chromosome. As with other Y. pestis PCR assays, three major genes located on each of the three virulence plasmids were included: lcrV on pCD1, caf1 on pMT1 and pla on pPCP1. The quadruplex assay was validated on a collection of 192 Y. pestis isolates and 52 near-neighbour isolates. It was discovered that only 72 % of natural plague isolates from the states of New Mexico and Utah harboured all three virulence plasmids. This quadruplex assay proved to be 100 % successful in differentiating Y. pestis from all near neighbours tested and was able to reveal which of the three virulence plasmids a particular isolate possessed. PMID- 18287296 TI - High-level carbapenem resistance in a Citrobacter freundii clinical isolate is due to a combination of KPC-2 production and decreased porin expression. AB - An imipenem-resistant isolate of Citrobacter freundii ZJ163 (MIC 256 microg ml( 1)) isolated from a Chinese hospital was investigated. The C. freundii ZJ163 isolate exhibited high-level resistance to carbapenems, penicillins, cephalosporins, cefoxitin, aztreonam, quinolones and aminoglycosides. Isoelectric focusing (IEF) demonstrated three beta-lactamases with pIs of 5.4 (TEM-1), 6.7 (KPC-2) and 7.9 (CTX-M-14). Two different transconjugants (types A and B) were obtained by conjugation studies. The type A transconjugant exhibited reduced susceptibility or resistance to penicillins, cephalosporins and aztreonam, but was susceptible to carbapenems, quinolones and aminoglycosides. The antimicrobial susceptibility patterns of the type B transconjugant were similar to that of type A, except for its significantly reduced carbapenem susceptibility (imipenem MIC 2 microg ml(-1)). IEF, specific PCRs and DNA sequence analysis indicated that the type A transconjugant produced CTX-M-14 beta-lactamase with a pI of 7.9, that the type B transconjugant produced KPC-2 beta-lactamase with a pI of 6.7 and that the beta-lactamase with a pI of 5.4 was TEM-1. PCR analysis and sequencing confirmed the presence of the ampC gene in the chromosomal DNA from C. freundii ZJ163, although no activity of AmpC beta-lactamase was detected by IEF. Urea/SDS-PAGE analysis of outer-membrane proteins revealed that the levels of the 41 and 38 kDa porins were decreased in C. freundii ZJ163. It was concluded that production of KPC-2 combined with decreased expression of porins contributes to high-level resistance to carbapenems in C. freundii ZJ163. PMID- 18287297 TI - PorA types in Neisseria meningitidis serogroup B isolated in Argentina from 2001 to 2003: implications for the design of an outer membrane protein-based vaccine. AB - Identification of Neisseria meningitidis PorA types remains important, as the PorA protein is a major immunogenic component of several meningococcal vaccines under development. In this study, 191 N. meningitidis serogroup B isolates collected in Argentina through active laboratory-based surveillance from 2001 to 2003 were serosubtyped. Nucleotide sequences of the porA variable region 1 (VR1) and VR2 regions were determined in 52 non-serosubtypeable isolates. A substantial number of distinct VR types were identified, and a new VR2 variant from the P1.16 family was described. This is the first report describing PorA types in N. meningitidis serogroup B isolates in Argentina. Furthermore, the wide diversity of subtypes detected by serosubtyping and genosubtyping reveals the difficulty in designing a useful outer-membrane vaccine applicable in this country. A possible mechanism responsible for altered PorA expression was analysed in two PorA types. PMID- 18287298 TI - Isolates of Clostridium perfringens recovered from Costa Rican patients with antibiotic-associated diarrhoea are mostly enterotoxin-negative and susceptible to first-choice antimicrobials. AB - To assess the prevalence of enterotoxigenic Clostridium perfringens among adults suffering from antibiotic-associated diarrhoea in a Costa Rican hospital, faecal samples were analysed from 104 patients by a cultivation approach. The 29 strains obtained, which accounted for an isolation frequency of 28 %, were genotyped and investigated with regard to their in vitro susceptibility to penicillin, imipenem, cefotaxime, chloramphenicol and metronidazole using an agar-dilution method. A multiplex PCR for detection of the toxins alpha, beta and epsilon predictably classified all faecal isolates as biotype A. An agglutination assay revealed that only one isolate synthesized detectable amounts of enterotoxin (detection rate 3 %). This result was confirmed by a PCR targeting the cpe gene. The spores of the only CPE(+) isolate did not germinate after incubation for 30 min at temperatures above 80 degrees C. Most isolates were susceptible to first choice antimicrobials. However, unusual MICs for penicillin (16 microg ml(-1)) and metronidazole (512 microg ml(-1)) were detected in one and three isolates, respectively. The low incidence of enterotoxigenic strains suggests that C. perfringens was not a major primary cause of antibiotic-associated diarrhoea in this hospital during the sampling period. PMID- 18287299 TI - Molecular typing of nasal carriage isolates of Staphylococcus aureus from an Irish university student population based on toxin gene PCR, agr locus types and multiple locus, variable number tandem repeat analysis. AB - Forty-eight Staphylococcus aureus isolates collected from a young, healthy, Irish university student population from 1995 to 2004 were screened for 16 enterotoxin (SE) and enterotoxin-like (SEl) genes (sea-see, seg-sei, selj-selo, selq, selu), and for the toxic shock toxin syndrome toxin-1 gene, tst. All of the isolates harboured at least one SE or SEl gene and 66.7 % possessed a classical SE gene (sea, seb, sec), the commonest being the seb gene. Most of the isolates (85.4 %) had a complete egc locus (selo, selm, sei, seln, seg). The intergenic sei-seln region of the egc locus was typed by PCR-RFLP in 34 isolates, 15 possessing pseudogenes psient1 and psient2 and 19 having the selu gene. The seh and sell genes, the selk-selq gene combination, and the tst gene were each found in <15 % of isolates. The agr genotype distribution was agr type III, 37.5 %; agr type I, 35.4 %; agr type II, 25 %; and agr type IV, 2.1 %. There was no association between SE-SEl genotype and agr type. All tst gene-positive isolates were of agr type III and harboured a classical SE gene. Multiple locus, variable number tandem repeat analysis (MLVA) produced 47 different patterns. While the sdr locus was present in all isolates, half of them lacked one or two of the sdr gene amplimers. Twenty isolates harboured the bbp gene, its presence being associated with agr type III, but not with the SE-SEl gene profile. The agr types of isolates were associated with MLVA subclusters. Selective MLST analysis revealed seven novel sequence types and a new aroE allele. Five clonal clusters (CCs), including CCs comprising major pandemic clones CC30, CC5 and CC22 and minor lineages CC6 and CC9, and three singletons were identified. PMID- 18287300 TI - Age-specific prevalence of diffusely adherent Escherichia coli in Brazilian children with acute diarrhoea. AB - In a prospective study between February 2003 and June 2004, stool specimens of children less than 2 years of age with diarrhoea (n=218) and without diarrhoea (n=86), living in Vitoria, Espirito Santo, Brazil, were examined for the presence of diarrhoeagenic Escherichia coli. E. coli isolates were tested by colony blot hybridization with specific DNA probes designed to detect EPEC, ETEC, EIEC, EAEC, DAEC and EHEC/STEC. Diarrhoeagenic E. coli strains were detected as the sole pathogen in stools of 92 (30.3 %) children, including 72 (33.0 %) with diarrhoea and 20 (23.2 %) without diarrhoea. DAEC was the most frequent pathotype and was found significantly more often from patients (18.3 %) than from controls (8.1 %) (P<0.05), particularly among children more than 1 year of age (P=0.01). Atypical EPEC and EAEC isolates were isolated from both patients (5.5 % and 4.6 %, respectively) and controls (6.9 % and 6.9 %, respectively). ETEC was more frequently isolated from patients (3.2 %) than controls (1.2 %). Typical EPEC (0.9 %) and EIEC (0.4 %) isolates were detected only in children with diarrhoea. In conclusion, our data suggest that DAEC should be considered potential pathogens in the region of Brazil studied. PMID- 18287301 TI - Analysis of meticillin-susceptible and meticillin-resistant biofilm-forming Staphylococcus aureus from catheter infections isolated in a large Italian hospital. AB - Several characteristics were analysed in 37 Staphylococcus aureus isolates from nosocomial catheter infections: the PFGE profile after SmaI digestion of chromosomal DNA, the ability to form a biofilm on a polystyrene surface, antibiotic susceptibility patterns (penicillin, oxacillin, erythromycin, tetracycline, clindamycin, telithromycin, gentamicin, ciprofloxacin, quinupristin/dalfopristin, rifampicin, vancomycin and linezolid), and the presence of genetic determinants of antibiotic resistance and biofilm formation. All strains but three (92 %) were able to grow on a plastic surface as a biofilm. An almost complete association was found between phenotypes and genotypic traits of antibiotic resistance, whilst PFGE profiling showed the highly polyclonal composition of the set of strains under study. Sixteen isolates (43 %) were meticillin-resistant and were subjected to staphylococcal cassette chromosome mec (SCCmec) and cassette chromosome recombinase (ccr) complex type determination by multiplex PCR. Only a subgroup of six strains belonged to the archaic clone PFGE type and bore the SCCmec/ccrAB type I structure. Among the remaining strains some presented small rearrangements of the SCCmec/ccrAB genetic locus, whilst others could barely be traced back to a known structural type. These observations suggest that, at the local level and at a particular site of infection, S. aureus may show great genetic variability and escape the general rule of expansion of the S. aureus pandemic clones. PMID- 18287302 TI - Streptococcus equi subsp. zooepidemicus meningitis. AB - A 72-year-old woman was hospitalized for Streptococcus equi subsp. zooepidemicus meningitis. The same organism was cultured from her two horses. She denied contact with horses, but had a practice of consuming unpasteurized milk from a cow. The cow was in the same stable as the horses, and the ill woman's son milked the cow. PMID- 18287303 TI - Neisseria elongata endocarditis complicated by brain embolism and abscess. AB - We report a case of Neisseria elongata endocarditis with thalamic septic embolization and subsequent brain abscess formation, which to the best of our knowledge has never been reported in the literature. The brain abscess completely resolved after a surgical repair of the infected mitral valve and an additional 4 weeks of antimicrobial therapy. Based on a review of all previous reports of N. elongata endocarditis, including ours, this will remind physicians that invasive N. elongata infections should be managed and followed up cautiously, as surgical intervention is often required. PMID- 18287304 TI - Stomatococcus mucilaginosus meningitis in a healthy 2-month-old child. AB - Stomatococcus mucilaginosus is a Gram-positive, catalase-variable organism considered part of the normal human oral and upper respiratory tract flora. Although traditionally believed to be an organism of low virulence, Stomatococcus mucilaginosus has been reported to be an opportunistic pathogen in immunocompromised patients. We describe what we believe is the first reported case of Stomatococcus mucilaginosus meningitis in a healthy child. The isolate was multidrug-resistant, susceptible only to vancomycin. The patient was treated successfully with vancomycin after initial trials with amikacin and cefotaxime. PMID- 18287305 TI - Legionnaires' disease in immunocompromised patients: a case report of Legionella longbeachae pneumonia and review of the literature. AB - In addition to Legionella pneumophila, about 20 Legionella species have been documented as human pathogens. The majority of infections by non-pneumophila Legionella species occur in immunocompromised and splenectomized patients. Here, we report a case of 'classical' lobar pneumonia caused by Legionella longbeachae in a splenectomized patient receiving corticosteroids for chronic immune thrombocytopenia. Tests for Legionella antigen were negative. L. longbeachae was immediately detected in bronchoalveolar fluid by PCR and subsequently confirmed by culture on legionella-selective media. The features of Legionnaires' disease in immunocompromised patients with special emphasis on significance and detection of non-pneumophila species are reviewed. PMID- 18287306 TI - Co-infection with two different strains of Bordetella pertussis in an infant. AB - We report co-infection with two phenotypically and genotypically distinct strains of Bordetella pertussis in an infant male hospitalized with a 2-week history of cough, paroxysms and vomiting. Colonies from the two B. pertussis phenotypes were isolated and evaluated by PFGE profile analysis, gene sequence typing and PCR RFLP of a portion of the 23S rRNA gene. These results demonstrated simultaneous infection with two different strains of B. pertussis. PMID- 18287307 TI - Identification of Acanthamoeba genotype T4 and Paravahlkampfia sp. from two clinical samples. AB - In this study, two free-living amoebae strains, Acanthamoeba genotype T4 and Paravahlkampfia sp., which were isolated from keratitis cases are presented. While the Acanthamoeba strain was isolated as a single agent, the Paravahlkampfia strain was found together with herpes simplex virus. Neither of the patients were contact lens wearers, but they did have a history of minor corneal trauma. Amoebae were detected on non-nutrient agar covered with Escherichia coli. Based on PCR-amplified 18S rRNA-gene analysis the first isolate was identified as Acanthamoeba genotype T4 and the second as Paravahlkampfia sp. In thermotolerance tests, the maximum temperature at which trophozoites continued to divide was determined as 37 degrees C for this Acanthamoeba strain and 35 degrees C for the Paravahlkampfia strain. To the best of our knowledge, the Acanthamoeba strain described herein is the second molecularly identified Acanthamoeba strain in an Acanthamoeba keratitis patient in Turkey. However, the Paravahlkampfia isolate is believed to be the first strain that has been isolated from a keratitis patient and has been molecularly differentiated from Vahlkampfia. PMID- 18287308 TI - Low recovery rates of high-level aminoglycoside-resistant enterococci could be attributable to restricted usage of aminoglycosides in Indian settings. PMID- 18287309 TI - Authors' reply to 'Misidentification of Bordetella bronchiseptica as Bordetella pertussis using a newly described RT-PCR targeting the pertactin gene'. PMID- 18287310 TI - Validation of virulence and epidemiology DNA microarray for identification and characterization of Staphylococcus aureus isolates. AB - The human pathogen Staphylococcus aureus is isolated and characterized using traditional culture and sensitivity methodologies that are slow and offer limited information on the organism. In contrast, DNA microarray technology can provide detailed, clinically relevant information on the isolate by detecting the presence or absence of a large number of virulence-associated genes simultaneously in a single assay. We have developed and validated a novel, cost effective multiwell microarray for the identification and characterization of Staphylococcus aureus. The array comprises 84 gene targets, including species specific, antibiotic resistance, toxin, and other virulence-associated genes, and is capable of examining 13 different isolates simultaneously, together with a reference control strain. Analysis of S. aureus isolates whose complete genome sequences have been determined (Mu50, N315, MW2, MRSA252, MSSA476) demonstrated that the array can reliably detect the combination of genes known to be present in these isolates. Characterization of a further 43 S. aureus isolates by the microarray and pulsed-field gel electrophoresis has demonstrated the ability of the array to differentiate between isolates representative of a spectrum of S. aureus types, including methicillin-susceptible, methicillin-resistant, community acquired, and vancomycin-resistant S. aureus, and to simultaneously detect clinically relevant virulence determinants. PMID- 18287311 TI - Supplementation of growth media with Zn2+ facilitates detection of VIM-2 producing Pseudomonas aeruginosa. PMID- 18287312 TI - Acute infantile gastroenteritis associated with human enteric viruses in Tunisia. AB - This prospective study, conducted from January 2003 to June 2005, investigated the incidence and the clinical role of various enteric viruses responsible for infantile gastroenteritis in 632 Tunisian children presenting in dispensaries (380 children) or hospitalized (252 children) for acute diarrhea. At least one enteric virus was found in each of 276 samples (43.7%). A single pathogen was observed in 234 samples, and mixed infections were found in 42 samples. In terms of frequency, rotavirus and norovirus were detected in 22.5 and 17.4% of the samples, respectively, followed by astrovirus (4.1%), Aichi virus (3.5%), adenovirus types 40 and 41 (2.7%), and sapovirus (1.0%). The seasonal distribution of viral gastroenteritis showed a winter peak but also an unusual peak from May to September. The severity of the diarrhea was evaluated for hospitalized infants. No significant differences were observed between rotavirus and norovirus infections with regard to the incidence and the clinical severity of the disease, especially in dehydration. PMID- 18287313 TI - Prevalence of Streptococcus invasive locus (sil) and its relationship with macrolide resistance among group A Streptococcus strains. PMID- 18287314 TI - Population structure of invasive and colonizing strains of Streptococcus agalactiae from neonates of six U.S. Academic Centers from 1995 to 1999. AB - The purpose of this study was to describe the population structure of group B streptococci (GBS) isolated from infected and colonized neonates during a prospective active-surveillance study of early-onset disease in six centers in the United States from July 1995 to June 1999 and to examine its relationship to bovine strains of GBS. The phylogenetic lineage of each GBS isolate was determined by multilocus sequence typing, and isolates were clustered into clonal complexes (CCs) using the eBURST software program. A total of 899 neonatal GBS isolates were studied, of which 129 were associated with invasive disease. Serotype Ia, Ib, and V isolates were highly clonal, with 92% to 96% of serotype Ia, Ib, and V isolates being confined to single clonal clusters. In contrast, serotype II and III isolates were each comprised of two major clones, with 39% of serotype II and 41% of serotype III isolates in CC 17 and 41% of serotype II and 54% of serotype III isolates in CC 19. Further analysis demonstrates that the CC 17 serotype II and III GBS are closely related to a previously described "ancestral" lineage of bovine GBS. While 120 (93%) of invasive GBS were confined to the same lineages that colonized neonates, 9 (7%) of the invasive GBS isolates were from rare lineages that comprised only 2.7% of colonizing lineages. These results are consistent with those for other geographic regions that demonstrate the highly clonal nature of GBS infecting and colonizing human neonates. PMID- 18287315 TI - Impact of human bocavirus on children and their families. AB - This study was planned to investigate the prevalence and clinical features of the illnesses associated with human bocavirus (hBoV) in children with acute disease. We prospectively enrolled all subjects aged less than 15 years attending an emergency room in Milan, Italy, on Wednesdays and Sundays between 1 November 2004 and 31 March 2005 for any acute medical reason, excluding surgical diseases and trauma. Nasopharyngeal swabs were collected at admission to detect hBoV; influenza A and B viruses; respiratory syncytial virus; human metapneumovirus; parainfluenza viruses 1, 2, 3, and 4; rhinovirus; adenovirus; and coronaviruses 229E, OC43, NL63, and HKU1 by real-time PCR. Among the 1,332 enrolled children, hBoV was the fifth most frequently detected virus (7.4%). The rate of hBoV coinfections with other viruses was significantly higher than for the other viruses (50.5% versus 27.5%; P < 0.0001). Eighty-nine of the 99 hBoV-positive children (89.9%) had a respiratory tract infection, and 10 (10.1%) had gastroenteritis. hBoV coinfections had a significantly greater clinical and socioeconomic impact on the infected children and their households than hBoV infection alone. In conclusion, these findings show that the role of hBoV infection alone seems marginal in children attending an emergency room for acute disease; its clinical and socioeconomic importance becomes relevant only when it is associated with other viruses. PMID- 18287316 TI - Fatal levofloxacin failure in treatment of a bacteremic patient infected with Streptococcus pneumoniae with a preexisting parC mutation. AB - The fatal outcome of levofloxacin treatment in a patient with bacteremic pneumonia caused by Streptococcus pneumoniae with a preexisting parC mutation is reported. Failure was due to the emergence of a gyrA mutation after 4 days of therapy. Problems encountered in detecting first-step mutation isolates are discussed. PMID- 18287317 TI - Prevalence of virulence genes and cytolethal distending toxin production in Campylobacter jejuni isolates from diarrheal patients in Bangladesh. AB - From 300 stool samples, 58 Campylobacter strains were isolated by standard microbiological and biochemical methods. Of these, 40 strains were identified as Campylobacter jejuni and 5 as Campylobacter coli. The presence of flaA (100%), cadF (100%), racR (100%), dnaJ (100%), pldA (100%), ciaB (95%), virB11 (0%), ceuE (82.5%), cdtA (97.5%), cdtB (97.5%), cdtC (97.5%), and wlaN (7.5%) genes was detected in C. jejuni by PCR. All C. jejuni strains but one produced cytolethal distending toxin in a HeLa cell assay. PMID- 18287318 TI - Increased sporulation rate of epidemic Clostridium difficile Type 027/NAP1. AB - Clostridium difficile PCR ribotype 027 comprised 0.2% of a collection of Swedish isolates in 1997-2001 (3 of 1,325 isolates). These isolates had lower moxifloxacin MICs than the epidemic type 027 isolates, but they had the same tcdC sequence and toxin yield. Type 027 produced 3- to 13-fold more toxin than did major Swedish types. One epidemic strain (027/NAP1a) sporulated more than did other type 027 isolates, a feature that should contribute to its survival and spread. PMID- 18287319 TI - Performance of the Cobas AmpliPrep/Cobas TaqMan real-time PCR assay for hepatitis B virus DNA quantification. AB - Hepatitis B virus (HBV) DNA quantification is used to establish the prognosis of chronic HBV-related liver disease, to identify those patients who need to be treated, and to monitor the virologic response and resistance to antiviral therapies. Real-time PCR-based assays are gradually replacing other technologies for routine quantification of HBV DNA in clinical practice. The goal of this study was to evaluate the intrinsic characteristics and clinical performance of the real-time PCR Cobas AmpliPrep/Cobas TaqMan (CAP/CTM) platform for HBV DNA quantification. Specificity was satisfactory (95% confidence interval, 98.1 to 100%). Intra-assay coefficients of variation ranged from 0.22% to 2.68%, and interassay coefficients of variation ranged from 1.31% to 4.13%. Quantification was linear over the full dynamic range of quantification of the assay (1.7 to 8.0 log(10) IU/ml) and was not affected by dilution. The assay was accurate regardless of the HBV genotype. Samples containing HBV DNA levels above 4.5 log(10) IU/ml were slightly underestimated relative to another accurate assay based on branched-DNA technology, but this is unlikely to have noteworthy clinical implications. Thus, the CAP/CTM HBV DNA assay is sensitive, specific, and reproducible, and it accurately quantifies HBV DNA levels in patients chronically infected by HBV genotypes A to F. Samples with HBV DNA concentrations above the upper limit of quantification need to be diluted and then retested. Broad use of fully automated real-time PCR assays should improve the management of patients with chronic HBV infection. PMID- 18287320 TI - Comparative analysis of human and canine Campylobacter upsaliensis isolates by amplified fragment length polymorphism. AB - Human (n = 33) and canine (n = 53) Campylobacter upsaliensis isolates from seven countries were genotyped by a new amplified fragment length polymorphism method. We observed 100% typeability and high overall diversity. The majority of human strains (23/33) clustered separately from canine strains, indicating that dogs may not be the main source of human infection. PMID- 18287321 TI - Comparison of reverse hybridization, microarray, and sequence analysis for genotyping hepatitis B virus. AB - Hepatitis B virus (HBV) genotyping has become important in epidemiological and clinical diagnoses, given the relationship between the viral genotype and the progression of disease or the appearance of antiviral resistance. Since genotyping by sequence and phylogenetic analyses is not convenient in the clinical setting, we evaluated InnoLipa HBV genotyping (Innogenetics, Belgium) and an HBV DNA-Chip (bioMerieux, France) prototype assay and compared their sequencing of the gold standard S gene, using a cohort of 275 individual patient samples. All but two samples, belonging to distant and individual subgroups within a single genotype, were detected by InnoLipa HBV assay. Four samples with dual infections belonging to genotypes A and G were identified only by InnoLipa HBV assay. Using an HBV DNA-Chip assay, one sample could not be amplified due to a low viral load. Four samples were identified as genotype C and two as genotype D by sequencing but were classified as genotype A (two samples) and D (two samples) and as A (one sample) and G (one sample) by the DNA-Chip assay. In conclusion, the InnoLipa HBV genotyping strip assay detected dual infections and was an easy and quick tool for genotyping, with a sensitivity of 99.3% and a specificity of 100% compared to sequence analysis. HBV DNA-Chip assay showed a sensitivity and specificity of 97.5 and 97.8%, respectively. PMID- 18287322 TI - Relationship between Mycobacterium tuberculosis genotype and the clinical phenotype of pulmonary and meningeal tuberculosis. AB - We used large sequence polymorphisms to determine the genotypes of 397 isolates of Mycobacterium tuberculosis from human immunodeficiency virus-uninfected Vietnamese adults with pulmonary (n = 235) or meningeal (n = 162) tuberculosis. We compared the pretreatment radiographic appearances of pulmonary tuberculosis and the presentation, response to treatment, and outcome of tuberculous meningitis between the genotypes. Multivariate analysis identified variables independently associated with genotype and outcome. A higher proportion of adults with pulmonary tuberculosis caused by the Euro-American genotype had consolidation on chest X-ray than was the case with disease caused by other genotypes (P = 0.006). Multivariate analysis revealed that meningitis caused by the East Asian/Beijing genotype was independently associated with a shorter duration of illness before presentation and fewer cerebrospinal fluid (CSF) leukocytes. Older age, fewer CSF leukocytes, and the presence of hemiplegia (but not strain lineage) were independently associated with death or severe disability, although the East Asian/Beijing genotype was strongly associated with drug-resistant tuberculosis. The genotype of M. tuberculosis influenced the presenting features of pulmonary and meningeal tuberculosis. The association between the East Asian/Beijing lineage and disease progression and CSF leukocyte count suggests the lineage may alter the presentation of meningitis by influencing the intracerebral inflammatory response. In addition, increased drug resistance among bacteria of the East Asian/Beijing lineage might influence the response to treatment. This study suggests the genetic diversity of M. tuberculosis has important clinical consequences. PMID- 18287323 TI - Prosthetic hip infection caused by Tropheryma whipplei. AB - We report a case of prosthetic hip infection due to Tropheryma whipplei in a 74 year-old man not previously known to have Whipple's disease. Diagnosis was based on systematic 16S rRNA gene amplification and sequencing of samples obtained during revision hip arthroplasty. PMID- 18287324 TI - Improved detection of circulating Aspergillus antigen by use of a modified pretreatment procedure. AB - Detection of circulating galactofuranose (galf) antigens, including galactomannan (GM), by the Platelia Aspergillus (PA) enzyme-linked immunosorbent assay (ELISA) is an important tool in the early diagnosis of invasive aspergillosis (IA). We used a modified pretreatment technique (MT) on consecutive negative PA ELISA plasma samples from IA patients in order to improve the detection of the fungal components present. Plasma samples (52) were collected from healthy donors, and 174 plasma samples with a galactomannan index (GMI) below 0.5 were collected from 25 unclassifiable and 23 IA patients. The PA ELISA reactivity of pretreated samples was determined before (conventional technique [CT]) and after (MT) filtration using a Microcon filter with a 50-kDa cutoff (Millipore). For the MT, the sensitivity of the PA ELISA increased from 42.9% (CT) to 78.6% (MT) using a cutoff for the GMI of 1.5 in the probable and proven group, whereas specificity slightly decreased from 98.7% to 96.1% in the control group. The 10-fold concentration step increased the GMI as high as 121-fold. The MT resulted not only in positive reactivity in samples that tested negative with the CT but also in the earlier detection of antigen by 2 to 17 days. PMID- 18287325 TI - Rapid and accurate identification of Candida albicans isolates by use of PNA FISHFlow. AB - We developed the simple, rapid (1 h), and accurate PNA FISH(Flow) method for the identification of Candida albicans. The method exploits unique in solution in situ hybridization conditions under which the cells are simultaneously fixed and hybridized. This method facilitates the accurate identification of clinical yeast isolates using two scoring techniques: flow cytometry and fluorescence microscopy. PMID- 18287326 TI - Etiological analysis of initial colonization of periodontal pathogens in oral cavity. AB - It is unclear when the initial colonization by periodontal pathogens occurs in the oral cavity. Therefore, we report here the association between specific age groups and the time when the initial colonization by periodontal pathogens occurs in the oral cavity in such groups. Findings are based on an epidemiological analysis of the prevalence of five periodontal pathogens in the oral cavities of a wide range of age populations, from newborn to elderly, who were randomly selected in a geographic region of Brazil. These periodontal pathogens include Campylobacter rectus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, and Tannerella forsythia and were analyzed in the bacterial samples isolated from gingival sulcus, the dorsum of the tongue, and cheek mucosa of diverse age groups, using a bacterial DNA specific PCR method. Results indicated that there are distinct age-related groups where initial colonization by the five periodontal pathogens examined in this study can be detected and that the presence of teeth is a permissive factor for colonization by P. gingivalis, P. intermedia, and T. forsythia. Although it remains unclear exactly how or when target pathogens colonize healthy subjects, an understanding of age-related groups does provide a potentially useful tool in the early detection and prevention of periodontitis in healthy individuals. PMID- 18287327 TI - The evolution of unintentional injury mortality among elderly in Europe. AB - OBJECTIVE: To compare cause-specific unintentional injury mortality trends among elderly (65+) in the European Union over a 10-year period. METHOD: Overall and cause-specific data for 23 out of the 29 EU and European Free Trade Association countries with population >/= 1,000,000 were retrieved from the World Health Organization (WHO), and age-standardized mortality rates for the first and last 3 available years of the study period were calculated. Proportional mortality changes were estimated through linear regression. RESULTS: Circa 1993, country specific rates varied widely (>fourfold), but this gap is closing and a statistically significant downward trend in overall mortality is noted circa 2002, in about half of the countries. Rates from falls were reduced by 4.3%, from motor vehicle traffic by 3.1%, and from smoke, fire, and flames by 3.1%. DISCUSSION: A large proportion of EU countries enjoys steady declining trends by major unintentional injury mortality category. Success factors and barriers underlying these benchmarking patterns should be further explored to accelerate the process of injury reduction. PMID- 18287328 TI - Self-care and professionally guided care in osteoarthritis: racial differences in a population-based sample. AB - OBJECTIVE: The aim of this study was to examine the prevalence of self-management practices among older White and African American persons with osteoarthritis. Self-management was defined broadly to include all behaviors adopted to reduce morbidity, whether recommended by physicians or not. METHODS: A population-based sample of Medicare beneficiaries (N = 551) was recruited. An expanded set of self management behaviors using structured and open-ended inquiry, along with use of arthritis-specific medications was elicited. RESULTS: Few differences in self care behaviors between race groups were found. However, older African American persons were significantly less likely to have prescriptions for nonsteroidal anti-inflammatory agents (NSAIDs) and more likely to use over-the-counter nonprescription analgesics. DISCUSSION: Older White and African American persons made similar use of self-care strategies to reduce disease morbidity. African Americans without access to prescription pain relievers substituted nonprescription analgesics. A broader view of self-management is valuable for assessing the ways people may move between professionally guided care and self care. PMID- 18287329 TI - The relationship of older adults' activities and body mass index. AB - OBJECTIVE: To investigate the relationship between Body Mass Index (BMI) and older adults' hours of participation in 31 activities. METHODS: Descriptive statistics are presented for the relationships between BMI and sociodemographics, health behaviors, and health characteristics. Linear regression was used to model the number of hours of participation in each activity. RESULTS: The hypothesis that being overweight or obese is associated with older adults' activities was supported. For example, compared to those of normal weight, obese older adults spend fewer hours walking, exercising, praying and meditating, house cleaning, and engaging in personal grooming. And, compared to normal weight older adults, obese older adults spend a greater number of hours watching television. DISCUSSION: These results substantiate prior findings of a negative relationship between physical activity and excess weight, and the lack of a relationship between social activity and BMI. PMID- 18287331 TI - Selective binding of RGMc/hemojuvelin, a key protein in systemic iron metabolism, to BMP-2 and neogenin. AB - Juvenile hemochromatosis is a severe and rapidly progressing hereditary disorder of iron overload, and it is caused primarily by defects in the gene encoding repulsive guidance molecule c/hemojuvelin (RGMc/HJV), a recently identified protein that undergoes a complicated biosynthetic pathway in muscle and liver, leading to cell membrane-linked single-chain and heterodimeric species, and two secreted single-chain isoforms. RGMc modulates expression of the hepatic iron regulatory factor, hepcidin, potentially through effects on signaling by the bone morphogenetic protein (BMP) family of soluble growth factors. To date, little is known about specific pathogenic defects in disease-causing RGMc/HJV proteins. Here we identify functional abnormalities in three juvenile hemochromatosis linked mutants. Using a combination of approaches, we first show that BMP-2 could interact in biochemical assays with single-chain RGMc species, and also could bind to cell-associated RGMc. Two mouse RGMc amino acid substitution mutants, D165E and G313V (corresponding to human D172E and G320V), also could bind BMP-2, but less effectively than wild-type RGMc, while G92V (human G99V) could not. In contrast, the membrane-spanning protein, neogenin, a receptor for the related molecule, RGMa, preferentially bound membrane-associated heterodimeric RGMc and was able to interact on cells only with wild-type RGMc and G92V. Our results show that different isoforms of RGMc/HJV may play unique physiological roles through defined interactions with distinct signaling proteins and demonstrate that, in some disease-linked RGMc mutants, these interactions are defective. PMID- 18287330 TI - VE-cadherin and beta-catenin binding dynamics during histamine-induced endothelial hyperpermeability. AB - Beta-catenin plays an important role in the regulation of vascular endothelial cell-cell adhesions and barrier function by linking the VE-cadherin junction complex to the cytoskeleton. The purpose of this study was to evaluate the effect of beta-catenin and VE-cadherin interactions on endothelial permeability during inflammatory stimulation by histamine. We first assessed the ability of a beta catenin binding polypeptide known as inhibitor of beta-catenin and T cell factor (ICAT) to compete beta-catenin binding to VE-cadherin in vitro. We then overexpressed recombinant FLAG-ICAT in human umbilical vein endothelial cells (HUVECs) to study its impact on endothelial barrier function controlled by cell cell adhesions. The binding of beta-catenin to VE-cadherin was quantified before and after stimulation with histamine along with measurements of transendothelial electrical resistance (TER) and apparent permeability to albumin (P(a)) under the same conditions. The results showed that ICAT bound to beta-catenin and competitively inhibited binding of the VE-cadherin cytoplasmic domain to beta catenin in a concentration-dependent manner. Overexpression of FLAG-ICAT in endothelial cell monolayers did not affect their basal permeability properties, as indicated by unaltered TER and P(a); however, the magnitude and duration of histamine-induced decreases in TER were significantly augmented. Likewise, the increase in P(a) in the presence of histamine was exacerbated. Overexpression of FLAG-ICAT also significantly decreased the level of beta-catenin-associated VE cadherin following histamine stimulation. Taken together, these data suggest that inflammatory agents like histamine cause a transient and reversible disruption of binding between beta-catenin and VE-cadherin, during which endothelial permeability is elevated. PMID- 18287332 TI - Role of extracellular superoxide in neutrophil activation: interactions between xanthine oxidase and TLR4 induce proinflammatory cytokine production. AB - Reactive oxygen species (ROS) contribute to neutrophil activation and the development of acute inflammatory processes in which neutrophils play a central role. However, there is only limited information concerning the mechanisms through which extracellular ROS, and particularly cell membrane-impermeable species, such as superoxide, enhance the proinflammatory properties of neutrophils. To address this issue, neutrophils were exposed to superoxide generating combinations of xanthine oxidase and hypoxanthine or lumazine. Extracellular superoxide generation induced nuclear translocation of nuclear factor-kappaB (NF-kappaB) and increased neutrophil production of the NF-kappaB dependent cytokines tumor necrosis factor-alpha (TNF-alpha) and macrophage inhibitory protein-2 (MIP-2). In contrast, there were no changes in TNF-alpha or MIP-2 expression when neutrophils lacking Toll-like receptor-4 (TLR4) were exposed to extracellular superoxide. Immunoprecipitation, confocal microscopy, and fluorescence resonance energy transfer (FRET) studies demonstrated association between TLR4 and xanthine oxidase. Exposure of neutrophils to heparin attenuated binding of xanthine oxidase to the cell surface as well as interactions with TLR4. Heparin also decreased xanthine oxidase-induced nuclear translocation of NF-kappaB as well as production of proinflammatory cytokines. These results demonstrate that extracellular superoxide has proinflammatory effects on neutrophils, predominantly acting through an TLR4-dependent mechanism that enhances nuclear translocation of NF-kappaB and increases expression of NF kappaB-dependent cytokines. PMID- 18287333 TI - A cystine-cysteine shuttle mediated by xCT facilitates cellular responses to S nitrosoalbumin. AB - We have shown previously that extracellular cysteine is necessary for cellular responses to S-nitrosoalbumin. In this study we have investigated mechanisms involved in accumulation of extracellular cysteine outside vascular smooth muscle cells and characterized the role of cystine-cysteine release in transfer of nitric oxide (NO)-bioactivity. Incubation of cells with cystine led to cystine uptake, reduction, and cysteine release. The process was inhibitable by extracellular glutamate, suggesting a role for system x(c)(-) amino acid transporters. Smooth muscle cells express this transporter constitutively and induction of the light chain component (xCT) by either diethyl maleate or 3 morpholino-sydnonimine (SIN-1) led to glutamate-inhibitable cystine uptake and an increased rate of cysteine release from cells. Likewise, overexpression of xCT in smooth muscle cells or endothelial cells led to glutamate-inhibitable cysteine release. The resulting extracellular cysteine was found to be required for transfer of NO from extracellular S-nitrosothiols into cells via system L transporters leading to formation of cellular S-nitrosothiols. Cysteine release coupled to cystine uptake was also found to be required for cellular responses to S-nitrosoalbumin and facilitated S-nitrosoalbumin-mediated inhibition of epidermal growth factor signaling. These data show that xCT expression can constitute a cystine-cysteine shuttle whereby cystine uptake drives cysteine release. Furthermore, we show that extracellular cysteine provided by this shuttle mechanism is necessary for transfer of NO equivalents and cellular responses to S-nitrosoablumin. PMID- 18287334 TI - Characterization of oxidized low-density lipoprotein-induced hormesis-like effects in osteoblastic cells. AB - Epidemiological studies indicate that patients suffering from atherosclerosis are predisposed to develop osteoporosis. Atherogenic determinants such as oxidized low-density lipoprotein (oxLDL) particles have been shown both to stimulate the proliferation and promote apoptosis of bone-forming osteoblasts. Given such opposite responses, we characterized the oxLDL-induced hormesis-like effects in osteoblasts. Biphasic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reductive activity responses were induced by oxLDL where low concentrations (10-50 microg/ml) increased and high concentrations (from 150 microg/ml) reduced the MTT activity. Cell proliferation stimulation by oxLDL partially accounted for the increased MTT activity. No alteration of mitochondria mass was noticed, whereas low concentrations of oxLDL induced mitochondria hyperpolarization and increased the cellular levels of reactive oxygen species (ROS). The oxLDL-induced MTT activity was not related to intracellular ROS levels. OxLDL increased NAD(P)H-associated cellular fluorescence and flavoenzyme inhibitor diphenyleneiodonium reduced basal and oxLDL-induced MTT activity, suggesting an enhancement of NAD(P)H-dependent cellular reduction potential. Low concentrations of oxLDL reduced cellular thiol content and increased metallothionein expression, suggesting the induction of compensatory mechanisms for the maintenance of cell redox state. These concentrations of oxLDL reduced osteoblast alkaline phosphatase activity and cell migration. Our results indicate that oxLDL particles cause hormesis-like response with the stimulation of both proliferation and cellular NAD(P)H-dependent reduction potential by low concentrations, whereas high concentrations lead to reduction of MTT activity associated with the cell death. Given the effects of low concentrations of oxLDL on osteoblast functions, oxLDL may contribute to the impairment of bone remodeling equilibrium. PMID- 18287335 TI - Expression and function of the rat vesicular monoamine transporter 2. AB - The vesicular monoamine transporters (VMATs) are essential proteins, involved in the storage of monoamines in the central nervous system and in endocrine cells, in a process that involves exchange of 2H(+) with one substrate molecule. The VMATs interact with various native substrates and clinically relevant drugs and display the pharmacological profile of multidrug transporters. Vesicular transporters suffer from a lack of biochemical and structural data due to the difficulties in their expression. In this work we present the high-level expression of rat VMAT2 (rVMAT2) in a stable a human embryonic kidney cell line (HEK293), generated using the resistance to the neurotoxin 1-methyl-4 phenylpyridinium (MPP(+)) conferred by the protein. In addition, we describe novel procedures for the solubilization and purification of active protein, and its reconstitution into proteoliposomes. The partially purified protein in detergent binds the inhibitor tetrabenazine and, after reconstitution, displays high levels of Deltamu(H+)-driven electrogenic transport of serotonin. The reconstituted purified rVMAT2 has wild-type affinity for serotonin, and its turnover rate is approximately 0.4 substrate molecule/s. PMID- 18287336 TI - Dynamics of single potassium channel proteins in the plasma membrane of migrating cells. AB - Cell migration is an important physiological process among others controlled by ion channel activity. Calcium-activated potassium channels (K(Ca)3.1) are required for optimal cell migration. Previously, we identified single human (h)K(Ca)3.1 channel proteins in the plasma membrane by means of quantum dot (QD) labeling. In the present study, we tracked single-channel proteins during migration to classify their dynamics in the plasma membrane of MDCK-F cells. Single hK(Ca)3.1 channels were visualized with QD- or Alexa488-conjugated antibodies and tracked at the basal cell membrane using time-lapse total internal reflection fluorescence (TIRF) microscopy. Analysis of the trajectories allowed the classification of channel dynamics. Channel tracks were compared with those of free QD-conjugated antibodies. The size of the label has a pronounced effect on hK(Ca)3.1 channel diffusion. QD-labeled channels have a (sub)diffusion coefficient D(QDbound) = 0.067 microm(2)/s(alpha), whereas that of Alexa488 labeled channels is D(Alexa) = 0.139 microm(2)/s. Free QD-conjugated antibodies move much faster: D(QDfree) = 2.163 microm(2)/s(alpha). Plotting the mean squared distances (msd) covered by hK(Ca)3.1 channels as a function of time points to the mode of diffusion. Alexa488-labeled channels diffuse normally, whereas the QD label renders hK(Ca)3.1 channel diffusion anomalous. Free QD-labeled antibodies also diffuse anomalously. Hence, QDs slow down diffusion of hK(Ca)3.1 channels and change the mode of diffusion. These results, referring to the role of label size and properties of the extracellular environment, suggest that the pericellular glycocalyx has an important impact on labels used for single molecule tracking. Thus tracking fluorescent particles within the glycocalyx opens up a possibility to characterize the pericellular nanoenvironment. PMID- 18287337 TI - Brain and leptomeningeal metastases from cutaneous melanoma: survival outcomes based on clinical features. AB - Brain metastases (BM) are among the most devastating and debilitating complications of melanoma. This retrospective study was conducted to gain a better understanding of patient and disease characteristics that have the greatest impact on overall survival in melanoma patients with BM; therapeutic interventions were also assessed. The records of all patients diagnosed with cutaneous melanoma and BM who were seen at Memorial Sloan-Kettering Cancer Center between 1991 and 2001 were retrospectively reviewed. A variety of factors, including age at diagnosis of stage IV disease, gender, race, disease stage at diagnosis, presence of BM at diagnosis of stage IV disease, neurologic symptoms, radiographic findings, number of BM, status and site(s) of extracranial metastasis, and treatment modalities, were analyzed for correlation with overall survival using univariate and multivariate Cox regression models. The records of 355 patients with BM were included in the analysis. On univariate analysis, seven patient and disease characteristics were significantly associated with poorer survival: age > 65 years, extracranial metastases, BM at stage IV diagnosis, neurologic symptoms, four or more BM, hydrocephalus, and leptomeningeal metastases. Of these, age, extracranial metastasis, neurologic symptoms, and number of BM were significantly associated with poorer survival in a multivariate analysis. Multivariate analysis of treatment modalities suggested that patients who had surgery, radiosurgery, or chemotherapy with temozolomide had improved survival outcomes, although this analysis has limitations. The prognostic factors identified in this retrospective study should be considered when making treatment decisions for patients with BM and used as stratification factors in future clinical trials. PMID- 18287338 TI - The utility of body FDG PET in staging primary central nervous system lymphoma. AB - (18)F-Fluorodeoxyglucose (FDG) PET has become an important tool in the management of non-Hodgkin's lymphoma (NHL), but its role in the evaluation of primary CNS lymphoma (PCNSL) has not been established. We investigated the ability of body FDG PET to detect systemic disease in the staging and restaging of PCNSL. The records of 166 PCNSL patients seen at Memorial Sloan-Kettering Cancer Center were examined. Forty-nine patients who underwent body FDG PET for staging of PCNSL were identified. Clinical data were reviewed to determine FDG PET results and their influence on therapy. Body FDG PET disclosed a systemic site of malignancy in 15% of patients. NHL was found in 11% of all patients, 7% of patients at diagnosis, and 27% of patients at CNS relapse. Four percent had a second systemic neoplasm. Workup with conventional staging did not reveal systemic disease, and in 8% of patients, body FDG PET was the only abnormal diagnostic exam suggestive of lymphoma. FDG PET findings altered patient treatment and resulted in additional chemotherapy, surgery, or radiotherapy. Our findings suggest that FDG PET may be more sensitive than conventional body staging and may disclose higher rates of concomitant systemic disease at PCNSL diagnosis. Body FDG PET may be an important noninvasive adjunct to conventional PCNSL staging, and its utility should be evaluated prospectively. PMID- 18287339 TI - A pilot study: 131I-antitenascin monoclonal antibody 81c6 to deliver a 44-Gy resection cavity boost. AB - The purpose of this study was to determine the feasibility and assess the efficacy and toxicity, among newly diagnosed malignant glioma patients, of administering (131)I-labeled murine antitenascin monoclonal antibody 81C6 ((131)I 81C6) into a surgically created resection cavity (SCRC) to achieve a patient specific, 44-Gy boost to the 2-cm SCRC margin. A radioactivity dose of (131)I 81C6 calculated to achieve a 44-Gy boost to the SCRC was administered, followed by conventional external beam radiotherapy (XRT) and chemotherapy. Twenty-one patients were enrolled in the study: 16 with glioblastoma multiforme (GBM) and 5 with anaplastic astrocytoma. Twenty patients received the targeted 44-Gy boost (+/-10%) to the SCRC. Attributable toxicity was mild and limited to reversible grade 3 neutropenia or thrombocytopenia (n = 3; 14%), CNS wound infections (n = 3; 14%), and headache (n = 2; 10%). With a median follow-up of 151 weeks, median overall survival times for all patients and those with GBM are 96.6 and 90.6 weeks, respectively; 87% of GBM patients are alive at 1 year. It is feasible to consistently achieve a 44-Gy boost dose to the SCRC margin with patient-specific dosing of (131)I-81C6. Our study regimen ((131)I-81C6 + XRT + temozolomide) was well tolerated and had encouraging survival. To determine if selection of good prognosis patients affects outcome associated with this approach, the U.S. Food and Drug Administration has approved a trial randomizing newly diagnosed GBM patients to either our study regimen or standard XRT plus temozolomide. PMID- 18287340 TI - Malignant pineal germ-cell tumors: an analysis of cases from three tumor registries. AB - The exact incidence of pineal germ-cell tumors is largely unknown. The tumors are rare, and the number of patients with these tumors, as reported in clinical series, has been limited. The goal of this study was to describe pineal germ-cell tumors in a large number of patients, using data from available brain tumor databases. Three different databases were used: Surveillance, Epidemiology, and End Results (SEER) database (1973-2001); Central Brain Tumor Registry of the United States (CBTRUS; 1997-2001); and National Cancer Data Base (NCDB; 1985 2003). Tumors were identified using the International Classification of Diseases for Oncology, third edition (ICD-O-3), site code C75.3, and categorized according to histology codes 9060-9085. Data were analyzed using SAS/STAT release 8.2, SEER*Stat version 5.2, and SPSS version 13.0 software. A total of 1,467 cases of malignant pineal germ-cell tumors were identified: 1,159 from NCDB, 196 from SEER, and 112 from CBTRUS. All three databases showed a male predominance for pineal germ-cell tumors (>90%), and >72% of patients were Caucasian. The peak number of cases occurred in the 10- to 14-year age group in the CBTRUS data and in the 15- to 19-year age group in the SEER and NCDB data, and declined significantly thereafter. The majority of tumors (73%-86%) were germinomas, and patients with germinomas had the highest survival rate (>79% at 5 years). Most patients were treated with surgical resection and radiation therapy or with radiation therapy alone. The number of patients included in this study exceeds that of any study published to date. The proportions of malignant pineal germ cell tumors and intracranial germ-cell tumors are in range with previous studies. Survival rates for malignant pineal germ-cell tumors are lower than results from recent treatment trials for intracranial germ-cell tumors, and patients that received radiation therapy in the treatment plan either with surgery or alone survived the longest. PMID- 18287341 TI - New therapeutic approach for brain tumors: Intranasal delivery of telomerase inhibitor GRN163. AB - The blood-brain barrier is a substantial obstacle for delivering anticancer agents to brain tumors, and new strategies for bypassing it are greatly needed for brain-tumor therapy. Intranasal delivery provides a practical, noninvasive method for delivering therapeutic agents to the brain and could provide an alternative to intravenous injection and convection-enhanced delivery. We treated rats bearing intracerebral human tumor xenografts intranasally with GRN163, an oligonucleotide N3'-->P5'thio-phosphoramidate telomerase inhibitor. 3'-Fuorescein isothiocyanate (FITC)-labeled GRN163 was administered intranasally every 2 min as 6 microl drops into alternating sides of the nasal cavity over 22 min. FITC labeled GRN163 was present in tumor cells at all time points studied, and accumulation of GRN163 peaked at 4 h after delivery. Moreover, GRN163 delivered intranasally, daily for 12 days, significantly prolonged the median survival from 35 days in the control group to 75.5 days in the GRN163-treated group. Thus, intranasal delivery of GRN163 readily bypassed the blood-brain barrier, exhibited favorable tumor uptake, and inhibited tumor growth, leading to a prolonged lifespan for treated rats compared to controls. This delivery approach appears to kill tumor cells selectively, and no toxic effects were noted in normal brain tissue. These data support further development of intranasal delivery of tumor specific therapeutic agents for brain tumor patients. PMID- 18287342 TI - Effect of phenytoin on celecoxib pharmacokinetics in patients with glioblastoma. AB - Cyclooxygenase-2 (COX-2) expression has been linked to the prognosis, angiogenesis, and radiation sensitivity of many malignancies. Celecoxib, a selective COX-2 inhibitor, is predominantly eliminated by hepatic metabolism. This study was conducted to determine the effects of hepatic enzyme-inducing antiseizure drugs (EIASDs) on the pharmacokinetics of celecoxib. The safety of celecoxib administered with radiation for glioblastoma and the effect of the combined treatment on survival were also evaluated. Patients were stratified based on concomitant use of EIASDs. Celecoxib (400) mg was administered orally twice a day until tumor progression or dose-limiting toxicity. Standard radiation was administered without adjuvant chemotherapy. Sampling was performed to define the plasma concentration/time profile for the initial dose of celecoxib and steady-state trough concentrations. Thirty-five patients (22 +EIASD, 13 -EIASD) were enrolled. There were no significant differences in age, performance status, extent of surgery, or Mini Mental State Exam scores between the two cohorts. The treatment was well tolerated. All patients in the +EIASD arm were taking phenytoin. There were no significant differences in any celecoxib pharmacokinetic parameters between 15 +EIASD and 12 -EIASD patients. With 31 of 35 patients deceased, estimated median survival time for all patients was 12 months (+EIASD, 11.5 months; - EIASD, 16 months; p = 0.11). The pharmacokinetics of celecoxib is not significantly affected by the concomitant administration of phenytoin. Celecoxib administered during and after radiation is well tolerated. The potential difference in survival between the +EIASD and -EIASD groups deserves further evaluation. PMID- 18287343 TI - Birth weight relates to salt sensitivity of blood pressure in healthy adults. AB - The association between birth weight and blood pressure is well established but at present unexplained. According to the Borst-Guyton concept, chronic hypertension can occur only with a shift in the renal pressure-natriuresis relationship resulting in increased salt sensitivity of blood pressure. We assessed salt sensitivity of blood pressure in a group of 27 healthy adults whose birth weight was available. Birth weight was ascertained from birth certificates or announcements. Salt sensitivity of blood pressure was determined as difference in mean arterial pressure (MAP) between a 1-week high-salt ( approximately 235 mmol NaCl/d) versus low-salt diet ( approximately 55 mmol NaCl/d). Creatinine clearance was estimated according to the formula of Cockcroft and Gault. Birth weight was negatively associated with salt sensitivity of blood pressure (r= 0.60, P=0.002). The creatinine clearance was positively associated with birth weight (r=0.53; P=0.008) but did not influence the association between birth weight and salt sensitivity of blood pressure. Birth weight is associated with salt sensitivity of blood pressure, and this may play a role in the maintenance of elevated blood pressure in individuals with a low birth weight. PMID- 18287344 TI - Are children doomed by what they eat and drink? PMID- 18287345 TI - Salt intake is related to soft drink consumption in children and adolescents: a link to obesity? AB - Dietary salt is a major determinant of fluid intake in adults; however, little is known about this relationship in children. Sugar-sweetened soft drink consumption is related to childhood obesity, but it is unclear whether there is a link between salt and sugar-sweetened soft drink consumption. We analyzed the data of a cross-sectional study, the National Diet and Nutrition Survey for young people in Great Britain. Salt intake and fluid intake were assessed in 1688 participants aged 4 to 18 years, using a 7-day dietary record. There was a significant association between salt intake and total fluid, as well as sugar-sweetened soft drink consumption (P<0.001), after adjusting for potential confounding factors. A difference of 1 g/d in salt intake was associated with a difference of 100 and 27 g/d in total fluid and sugar-sweetened soft drink consumption, respectively. These results, in conjunction with other evidence, particularly that from experimental studies where only salt intake was changed, demonstrate that salt is a major determinant of fluid and sugar-sweetened soft drink consumption during childhood. If salt intake in children in the United Kingdom was reduced by half (mean decrease: 3 g/d), there would be an average reduction of approximately 2.3 sugar-sweetened soft drinks per week per child. A reduction in salt intake could, therefore, play a role in helping to reduce childhood obesity through its effect on sugar-sweetened soft drink consumption. This would have a beneficial effect on preventing cardiovascular disease independent of and additive to the effect of salt reduction on blood pressure. PMID- 18287346 TI - Metabolic effects of dietary fiber consumption and prevention of diabetes. AB - A high dietary fiber (DF) intake is emphasized in the recommendations of most diabetes and nutritional associations. It is accepted that viscous and gel forming properties of soluble DF inhibit macronutrient absorption, reduce postprandial glucose response, and beneficially influence certain blood lipids. Colonic fermentation of naturally available high fiber foods can also be mainly attributed to soluble DF, whereas no difference between soluble and insoluble DF consumption on the regulation of body weight has been observed. However, in prospective cohort studies, it is primarily insoluble cereal DF and whole grains, and not soluble DF, that is consistently associated with reduced diabetes risk, suggesting that further, unknown mechanisms are likely to be involved. Recent research indicates that DF consumption contributes to a number of unexpected metabolic effects independent from changes in body weight, which include improvement of insulin sensitivity, modulation of the secretion of certain gut hormones, and effects on various metabolic and inflammatory markers that are associated with the metabolic syndrome. In this review, we briefly summarize novel findings from recent interventions and prospective cohort studies. We discuss concepts and potential mechanisms that might contribute to the further understanding of the involved processes. PMID- 18287347 TI - Oleanolic acid induces prostacyclin release in human vascular smooth muscle cells through a cyclooxygenase-2-dependent mechanism. AB - Oleanolic acid is a triterpenoid that may contribute to the cardio-protective effects of olive oil. Our goal was to assess whether oleanolic acid could modulate eicosanoid biosynthesis and to determine the mechanism involved in this effect. Human coronary smooth muscle cells (SMC) were treated with oleanolic acid, erythrodiol, or hydroxytyrosol and eicosanoid release was measured by enzyme immunoassay. Cyclooxygenase (Cox)-1 and Cox-2 protein and messenger sRNA levels were analyzed by Western blot and real-time PCR, respectively. Mitogen activated protein kinase (MAPK) pathways were assessed using specific antibodies. Oleanolic acid induced prostaglandin I2 (PGI2) release by human coronary SMC, an effect that was prevented by celecoxib (a specific inhibitor of Cox-2). The increased PGI2 was time-and dose-dependent and was associated to the up regulation of Cox-2. No effects were observed on thromboxane A2. Erythrodiol but not hydroxytyrosol upregulated Cox-2 expression and induced PGI2 synthesis. Oleanolic acid induced an early phosphorylation of p38 MAPK and p42/44 MAPK but not c-Jun N-terminal kinase-1 (JNK-1). SB203580 (p38MAPK inhibitor) and U0126 (MAPK kinase1/2 inhibitor) abrogated the upregulation of Cox-2 and PGI2 release induced by oleanolic acid. A peptide inhibitor of JNK-1 (L-JNKI1) did not produce any effect. The induction of Cox-2 was preceded by an early activation of cAMP regulatory element-binding protein, a key transcription factor involved in Cox-2 transcriptional upregulation. Therefore, oleanolic acid contributes to vascular homeostasis by inducing PGI2 release in a Cox-2-dependent manner. Oleanolic acid could be regarded as a bioactive molecule that may contribute to the beneficial effects of the Mediterranean diet. PMID- 18287348 TI - Conjugated linoleic acid reduces body fat accretion and lipogenic gene expression in neonatal pigs fed low- or high-fat formulas. AB - Childhood obesity is an increasing problem and may predispose children to adult obesity. Weight gain during infancy has been linked to excessive weight later in life. Conjugated linoleic acids (CLA) have been shown to reduce fat gain and body fat mass in animal models and in humans. The effects of CLA in a piglet model of human infancy have not been determined. The objective of this experiment was to examine the regulation of body composition and lipid metabolism in pigs fed low- and high-fat milk formulas supplemented with CLA. Twenty-four piglets were fed low- (3%) or high-fat (25%) diets with or without 1% CLA in a 2 x 2 factorial design. Formulas were fed for 16-17 d. Piglet body weight gains did not differ, although pigs fed the low-fat diets consumed greater amounts of diet. Piglets fed the high-fat formula accreted 50% more body fat during the feeding period than low-fat fed piglets and CLA reduced body fat accretion regardless of dietary fat content. Liver and muscle in vitro oxidation of palmitate was not influenced by dietary treatments. Adipose tissue expression of acetyl-CoA carboxylase-alpha and lipoprotein lipase were significantly reduced by CLA treatment. Overall, CLA reduced body fat accretion without influencing daily gain in a piglet model of human infancy. Results indicate that inhibition of fatty acid uptake and synthesis by adipose tissue, and not increased fatty acid oxidation in liver or muscle, were involved in reducing body fat gain. PMID- 18287349 TI - Trans-10, cis-12 conjugated linoleic acid antagonizes ligand-dependent PPARgamma activity in primary cultures of human adipocytes. AB - We previously demonstrated that trans-10, cis-12 (10,12) conjugated linoleic acid (CLA) causes human adipocyte delipidation, insulin resistance, and inflammation in part by attenuating PPARgamma target gene expression. We hypothesized that CLA antagonizes the activity of PPARgamma in an isomer-specific manner. 10,12 CLA, but not cis-9, trans-11 (9,11) CLA, suppressed ligand-stimulated activation of a peroxisome proliferator response element-luciferase reporter. This decreased activation of PPARgamma by 10,12 CLA was accompanied by an increase in PPARgamma and extracellular signal-related kinase (ERK)1/2 phosphorylation, followed by decreased PPARgamma protein levels. To investigate if 10,12 CLA-mediated delipidation was preventable with a PPARgamma ligand (BRL), cultures were treated for 1 wk with 10,12 CLA or 10,12 CLA + BRL and adipogenic gene and protein expression, glucose uptake, and triglyceride (TG) were measured. BRL cosupplementation completely prevented 10,12 CLA suppression of adipocyte fatty acid-binding protein, lipoprotein lipase, and perilipin mRNA levels without preventing reductions in PPARgamma or insulin-dependent glucose transporter 4 (GLUT4) expression, glucose uptake, or TG. Lastly, we investigated the impact of CLA withdrawal in the absence or presence of BRL for 2 wk. CLA withdrawal did not rescue CLA-mediated reductions in adipogenic gene and protein expression. In contrast, BRL supplementation for 2 wk following CLA withdrawal rescued mRNA levels of PPARgamma target genes. However, the levels of PPARgamma and GLUT4 protein and TG were only partially rescued by BRL. Collectively, we demonstrate for the first time, to our knowledge, that 10,12 CLA antagonizes ligand-dependent PPARgamma activity, possibly via PPARgamma phosphorylation by ERK. PMID- 18287350 TI - Soy protein ameliorates metabolic abnormalities in liver and adipose tissue of rats fed a high fat diet. AB - Chronic consumption of high-fat or -carbohydrate diets is associated with the development of obesity; however, it is not well established whether dietary protein plays a role in the development of abnormalities of lipid metabolism that occur during obesity. To determine the effect of different types of protein during diet-induced obesity on hepatic and adipocyte lipid metabolism, rats were fed casein (CAS) or soy (SOY) protein diets with 5% fat or high-fat diets with 25% fat (HF-CAS and HF-SOY) for 180 d. Rats fed soy diets had lower hepatic sterol regulatory element binding protein-1 (SREBP-1) expression and higher SREBP 2 expression than those fed casein diets, leading to less hepatic lipid deposition. On the other hand, long-term HF-SOY consumption prevented hyperleptinemia in comparison with rats fed HF-CAS. Rats fed soy protein diet showed higher adipocyte perilipin mRNA expression and smaller adipocyte area than those fed casein diets, which was associated with a lower body fat content. Furthermore, the lipid droplet area in brown adipose tissue was significantly lower in rats fed soy diets than in those fed casein diets and it was associated with higher uncoupling protein-1 (UCP-1) expression. As a result, rats fed the soy diets gained less weight than those fed the casein diets, in part due to an increase in the thermogenic capacity mediated by UCP-1. These results suggest that the type of protein consumed and the presence of fat in the diet modulate lipid metabolism in adipose tissue and liver. PMID- 18287351 TI - Carrageenan induces cell cycle arrest in human intestinal epithelial cells in vitro. AB - Multiple studies in animal models have shown that the commonly used food additive carrageenan (CGN) induces inflammation and intestinal neoplasia. We performed the first studies to determine the effects of CGN exposure on human intestinal epithelial cells (IEC) in tissue culture and tested the effect of very low concentrations (1-10 mg/L) of undegraded, high-molecular weight CGN. These concentrations of CGN are less than the anticipated exposure of the human colon to CGN from the average Western diet. In the human colonic epithelial cell line NCM460 and in primary human colonic epithelial cells that were exposed to CGN for 1-8 d, we found increased cell death, reduced cell proliferation, and cell cycle arrest compared with unexposed control cells. After 6-8 d of CGN exposure, the percentage of cells reentering G0-G1 significantly decreased and the percentages of cells in S and G2-M phases significantly increased. Increases in activated p53, p21, and p15 followed CGN exposure, consistent with CGN-induced cell cycle arrest. Additional data, including DNA ladder, poly ADP ribose polymerase Western blot, nuclear DNA staining, and activities of caspases 3 and 7, indicated no evidence of increased apoptosis following CGN exposure and were consistent with CGN-induced necrotic cell death. These data document for the first time, to our knowledge, marked adverse effects of low concentrations of CGN on survival of normal human IEC and suggest that CGN exposure may have a role in development of human intestinal pathology. PMID- 18287352 TI - Lipid extract of Nostoc commune var. sphaeroides Kutzing, a blue-green alga, inhibits the activation of sterol regulatory element binding proteins in HepG2 cells. AB - Nostoc commune var. sphaeroides Kutzing (N. commune), a blue-green alga, has been used as both a food ingredient and in medicine for centuries. To determine the effect of N. commune on cholesterol metabolism, N. commune lipid extract was incubated at increasing concentrations (25-100 mg/L) with HepG2 cells, a human hepatoma cell line. The addition of N. commune lipid extract markedly reduced mRNA abundance of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and LDL receptor (LDLR) (P < 0.05), with a concomitant decrease in their protein expression (P < 0.001). Reduced HMGR activity by 90% with N. commune lipid extract confirmed the inhibitory role of N. commune in cholesterol synthesis (P < 0.006). To elucidate a molecular mechanism underlying the repression of HMGR and LDLR by N. commune lipid extract, expression of sterol regulatory element binding protein 2 (SREBP-2) was assessed. Whereas mRNA for SREBP-2 remained unchanged, SREBP-2 mature protein was reduced by N. commune (P < 0.009). In addition, N. commune lipid extract also decreased SREBP-1 mature protein by approximately 30% (P < 0.002) and reduced the expression of SREBP-1-responsive genes such as fatty acid synthase and stearoyl CoA desaturase 1 (SCD-1) (P < 0.05). Therefore, our results demonstrate that N. commune lipid extract inhibits the maturation process of both SREBP-1 and -2, resulting in a decrease in expression of genes involved in cholesterol and fatty acid metabolism. PMID- 18287353 TI - In vitro micellarization and intestinal cell uptake of cis isomers of lycopene exceed those of all-trans lycopene. AB - The ratio of cis and all-trans lycopene (LYC) in human and animal tissues exceeds that in foods. The basis for this difference remains unknown, although differences in their stability, transport, and metabolism have been suggested. Here, we systematically compared the digestive stability, efficiency of micellarization, and uptake and intracellular stability of cis and all-trans isomers of LYC and carotenes using the coupled in vitro digestion and Caco-2 human intestinal cell model. Aril and oil from the carotenoid-rich gac fruit (Momordica cochinchinensis Spreng) were cooked with rice to provide a natural source of LYC and carotenes. The ratio of cis:trans isomers of LYC and beta carotene was similar before and after simulated gastric and small intestinal digestion with recovery of total carotenoids in the digesta exceeding 70%. Micellarization of cis isomers of LYC during digestion of meals with both gac aril and oil was significantly greater than that of the all-trans isomer but less than for the carotenes. Uptake of cis isomers of LYC by Caco-2 cells was similar to that of carotenes and significantly greater than all-trans LYC. Micellarized carotenoids were relatively stable in micelles incubated in the cell culture environment and after accumulation in Caco-2 cells. These data suggest that the greater bioaccessibility of cis compared with all-trans isomers of LYC contributes to the enrichment of the cis isomers in tissues and that gac fruit is an excellent source of bioaccessible LYC and provitamin A carotenoids. PMID- 18287354 TI - Feeding status regulates the polyubiquitination step of the ubiquitin-proteasome dependent proteolysis in rainbow trout (Oncorhynchus mykiss) muscle. AB - In mammals, the ubiquitin-proteasome proteolytic pathway is a major route of protein degradation and has been shown to be regulated by the feeding status via the protein kinase B (PKB)-Forkehead box-O transcription factor signaling pathway mediated transcription regulation of atrophy-related ubiquitin ligases, atrogin1 and muscle RING finger 1. In contrast, in rainbow trout (Oncorhynchus mykiss), the activity of the proteasome in muscle was not affected during starvation induced muscle degradation. The aim of this study was therefore to explore the molecular basis for this lack of induction of this proteolytic route during starvation. In this study, rainbow trout were food deprived for 7 and 14 d, refed ad libitum, and the effect of the nutritional status was assessed on the different steps involved in the regulation of the ubiquitin-proteasome system in muscle. We observed that starvation reduced the phosphorylation of PKB and enhanced the expression of atrogin1 in muscle, whereas refeeding led to the opposite effects. The level of polyubiquitinated proteins in muscle increased to over 2 times the initial value on d 0 after 14 d of starvation and decreased significantly at 12 h after refeeding, but there were no major changes in the activity of the main proteasomal peptidases (chymotrypsin-like and trypsin-like). Altogether, these results indicate that in rainbow trout muscle, the polyubiquitination step of the ubiquitin-proteasome route is regulated by the feeding status similarly to what is observed in mammals. PMID- 18287355 TI - Age and dietary form of vitamin K affect menaquinone-4 concentrations in male Fischer 344 rats. AB - Phylloquinone, the primary dietary form of vitamin K, is converted to menaquinone 4 (MK-4) in certain tissues. MK-4 may have tissue-specific roles independent of those traditionally identified with vitamin K. Fischer 344 male rats of different ages (2, 12, and 24 mo, n = 20 per age group) were used to compare the conversion of phylloquinone to MK-4 with an equivalent dose of another dietary form of vitamin K, 2',3'-dihydrophylloquinone. Rats were age- and diet-group pair-fed phylloquinone (198 +/- 9.0 microg/kg diet) or dihydrophylloquinone (172 +/- 13.0 microg/kg diet) for 28 d. MK-4 was the primary form of vitamin K in serum, spleen, kidney, testes, bone marrow, and brain myelin fractions, regardless of age group. MK-4 concentrations were significantly lower in kidney, heart, testes, cortex (myelin), and striatum (myelin) in the dihydrophylloquinone diet group compared with the phylloquinone diet group (P < 0.05). The MK-4 concentrations in 2-mo-old rats were lower in liver, spleen, kidney, heart, and cortex (myelin) but higher in testes compared with 24-mo-old rats (P < 0.05). However, there were no age-specific differences in MK-4 concentrations among the rats fed the 2 diets. These data suggest that dihydrophylloquinone, which differs from phylloquinone in its side phytyl chain, is absorbed but its intake results in less MK-4 in certain tissues. Dihydrophylloquinone may be used in models for the study of tissue specific vitamin K deficiency. PMID- 18287356 TI - Mammalian sirtuin 1 is involved in the protective action of dietary saturated fat against alcoholic fatty liver in mice. AB - This study was undertaken to elucidate the mechanism underlying the protective effect of a high saturated fat (HSF) diet against the development of alcoholic fatty liver in mice. We tested the effects of a HSF diet on the ethanol-mediated increase in hepatic sterol regulatory element binding protein 1 (SREBP-1) activity. Thirty-two male mice were divided into 4 groups and fed liquid diets consisting of either a high polyunsaturated fat (40% of energy from corn oil) or a HSF (40% of energy from cocoa butter) diet with or without ethanol for 4 wk. In the ethanol-containing diets, ethanol was substituted for an equivalent amount of carbohydrate to provide 27.5% of the total energy. Control mice were pair-fed the same volume of liquid diets as the ethanol-fed mice. The HSF diet suppressed the increase in mature SREBP-1 protein and prevented increased mRNA of the SREBP-1 regulated lipogenic enzymes in the ethanol-fed mice (P < 0.05). Sirtuins 1 (SIRT1), a NAD+-dependent class III histone deacetylase, was upregulated by ethanol administration in mice fed the HSF diet (P < 0.05). The HSF diet blocked histone H3 at lysine 9 (lys9) hyperacetylation and attenuated association of acetylated histone H3-Lys9 with the promoters of mitochondrial glycerol-3 phosphate acyltransferase and stearoyl-CoA desaturase 1 in the livers of the ethanol-fed mice. These results suggest that the protective effects of HSF diet against the development of alcoholic liver steatosis may occur via regulation of the hepatic SIRT1-SREBP-1-histone H3 axis, suppressing the expression of genes encoding lipogenic enzymes and slowing the synthesis of hepatic fatty acids. PMID- 18287357 TI - Nonstarch polysaccharide hydrolysis products of soybean and canola meal protect against enterotoxigenic Escherichia coli in piglets. AB - Infectious diarrhea is a major problem in both children and piglets. Enterotoxigenic Escherichia coli (ETEC) infection results in fluid and electrolyte losses in the small intestine. We investigated the effect of nonstarch polysaccharide (NSP) hydrolysis products of soybean meal (SBM) and canola meal (CM) on net absorption of fluid and solutes during ETEC infection. Products were generated by incubating SBM and CM with a blend of carbohydrase enzymes. Following incubation, slurries were centrifuged and the supernatants mixed with absolute ethanol to produce 2 product types: 80% ethanol-soluble (ES) and 80% ethanol-insoluble (EI). Products from SBM and CM were studied in 2 independent experiments in which 2 factors were investigated: product type (EI vs. ES) and time of ETEC infection (before vs. after perfusion). Pairs of small intestine segments, one noninfected and the other ETEC infected, were perfused simultaneously with different products for 7.5 h. Net absorption of fluid and solutes were determined. In both experiments, ETEC-infected segments perfused with saline control had lower (P < or = 0.05) net fluid and solute absorption compared with SBM and CM products. The interaction (P < or = 0.05) between product type and time of infection on fluid absorption was only evident for SBM, in which case perfusing ES products before infection resulted in higher fluid absorption (735 +/- 22 microL/cm2) compared with ETEC infection before perfusion (428 +/- 34 microL/cm2). In conclusion, NSP hydrolysis products of SBM and CM, particularly ES from SBM, were beneficial in maintaining fluid balance during ETEC infection, suggesting potential for controlling ETEC-induced diarrhea in piglets. PMID- 18287358 TI - A diet rich in conjugated linoleic acid and butter increases lipid peroxidation but does not affect atherosclerotic, inflammatory, or diabetic risk markers in healthy young men. AB - Intake of conjugated linoleic acid (CLA) has been demonstrated to beneficially affect risk markers of atherosclerosis and diabetes in rats. CLA is naturally found in milk fat, especially from cows fed a diet high in oleic acid, and increased CLA intake can occur concomitantly with increased milk fat intake. Our objective was to investigate the effect of CLA as part of a diet rich in butter as a source of milk fat on risk markers of atherosclerosis, inflammation, diabetes type II, and lipid peroxidation. A total of 38 healthy young men were given a diet with 115 g/d of CLA-rich fat (5.5 g/d CLA oil, a mixture of 39.4% cis9, trans11 and 38.5% trans10, cis12) or of control fat with a low content of CLA in a 5-wk double-blind, randomized, parallel intervention study. We collected blood and urine before and after the intervention. The fatty acid composition of plasma triacylglycerol, cholesterol esters, and phospholipids reflected that of the intervention diets. The CLA diet resulted in increased lipid peroxidation measured as an 83% higher 8-iso-prostaglandin F2alpha concentration compared with the control, P < 0.0001. We observed no other significant differences in the effect of the interventions diets. In conclusion, when given as part of a diet rich in butter, a mixture of CLA isomers increased lipid peroxidation but did not affect risk markers of cardiovascular disease, inflammation, or fasting insulin and glucose concentrations. PMID- 18287359 TI - Lipid metabolism in diet-induced obese rabbits is similar to that of obese humans. AB - Whereas diet-induced obese rabbits have been used to study various aspects of obesity, alterations of lipid metabolism in this model have not been clarified. This study aimed to compare plasma nonesterified fatty acid (NEFA) and triglyceride (TG) kinetics in obese and lean rabbits by means of U-(13)C16 palmitate infusion. Young female rabbits consumed either a high-fat diet (49% energy from fat) ad libitum to develop obesity (n = 6) or a normal diet (7.9% energy from fat) as lean control (n = 5). After 10 wk of feeding, the body weight of obese rabbits (5.33 +/- 0.05 kg) was greater (P < 0.001) than that of lean rabbits (3.89 +/- 0.07 kg). The obese rabbits had higher concentrations of plasma NEFA and TG and a greater rate of fatty acid (FA) turnover. Whereas the fractional secretion rates of hepatic TG did not differ, 100% of hepatic secretory TG was synthesized from plasma NEFA in the lean rabbits compared to 59% in the obese rabbits (P < 0.001). In the lean rabbits, hepatic lipase-mediated hydrolysis of lipoprotein TG did not contribute to the FA pool for synthesis of secretory TG, consistent with the naturally occurring deficit in hepatic lipase in this species. We conclude that lipid metabolism in diet-induced obese rabbits is similar to that in obese humans. The deficiency in hepatic lipase in rabbits simplifies the quantitation of hepatic lipid kinetics. PMID- 18287360 TI - Chronic treatment with red wine polyphenol compounds mediates neuroprotection in a rat model of ischemic cerebral stroke. AB - In this study, we investigated the in vivo effects of red wine polyphenol compounds (RWPC) in rats that were submitted to middle cerebral occlusion as an experimental model of stroke. Male Wistar rats were given RWPC [30 mg/(kg x d) dissolved in drinking water] or water for 1 wk before being subjected to transient middle cerebral artery occlusion followed by reperfusion. Sham-operated rats were subjected to transient occlusion in which the filament was not completely introduced. The release of amino acids and energy metabolites were monitored by intracerebral microdialysis. The volume of the ischemic lesion was assessed 24 h after reperfusion. Proteomic analysis of brain tissue was performed to study the effects of ischemia and RWPC on specific protein expression. Treatment with RWPC completely prevented the burst of excitatory amino acids that occurred in response to ischemia in untreated rats and significantly reduced brain infarct volumes. Rats chronically treated with RWPC, however, had lower basal concentrations of energy metabolites, including glucose and lactate in the brain parenchyma, compared with untreated rats. Chronic RWPC treatment significantly enhanced the residual cerebral blood flow during occlusion and reperfusion in rats subjected to transient occlusion compared with untreated rats. This effect resulted from arterial vasodilatation, as the internal diameters of several arteries were significantly enlarged after RWPC treatment. Proteomic studies revealed the modulation by RWPC of the expression of proteins involved in the maintenance of neuronal caliber and axon formation, in the protection against oxidative stress, and in energy metabolism. These findings provide an experimental basis for the beneficial effects of RWPC on the neurovascular unit during stroke. PMID- 18287361 TI - Cruciferous vegetables reduce morphological markers of colon cancer risk in dimethylhydrazine-treated rats. AB - Consumption of cruciferous vegetables has been associated with reduced colon cancer risk in human populations. However, little experimental evidence exists to support this association. Here, we report the effects of diets containing cruciferous vegetables on colon cancer risk. In Expt. 1, rats were fed a vegetable-free (basal) diet or diets containing different lyophilized cruciferous vegetables in concentrations between 4 and 10%. In Expt. 2, rats were fed the basal diet or diets containing 10-22.6% fresh cruciferous vegetables. Diets were fed for 2 wk (Expt. 1) or 3 wk (Expt. 2) before and 7 wk (Expt. 1) or 12 wk (Expt. 2) after administration of the colon carcinogen 1,2-dimethylhydrazine. Rats fed fresh vegetables were also injected with a low dose of carcinogen 18-24 h prior to termination. Groups fed lyophilized vegetables did not differ in aberrant crypt foci (ACF), sialomucin-producing foci, or mucin-depleted foci (MDF) numbers. However, all fresh vegetable diets significantly decreased ACF (approximately 40%) and MDF numbers. Activities of the hepatic phase I enzyme CYP2E1 did not differ among groups in either experiment. Hepatic glutathione S transferase (GST) and quinone reductase activities did not differ among groups fed fresh vegetables, whereas the lyophilized cabbage diets decreased GST activity compared with the basal diet. Groups did not differ in apoptosis and cell proliferation labeling indices in colonic mucosa. This study indicates that fresh but not lyophilized cruciferous vegetables reduce colon cancer risk in rats. These results do not support changes in hepatic carcinogen metabolism or colonic crypt cytokinetics as a mechanism. PMID- 18287362 TI - Dietary plasma proteins modulate the immune response of diffuse gut-associated lymphoid tissue in rats challenged with Staphylococcus aureus enterotoxin B. AB - We have previously shown that plasma protein supplementation prevents the activation of lymphocyte populations of Peyer's patches and mesenteric lymph nodes, which is known as organized gut-associated lymphoid tissue (GALT). Here, we examined the effects of spray-dried plasma proteins (SDAP) and Ig concentrate (IgC) supplements on lamina propria and intraepithelial lymphocytes (diffuse GALT) in a model of mild intestinal inflammation induced by the intraperitoneal administration of Staphylococcus aureus enterotoxin B (SEB). Wistar-Lewis rats were fed diets supplemented with SDAP (8% wt:wt), IgC (1.5% wt:wt), or bovine milk proteins (control diet) from weaning (d 21) to d 34 after birth. On d 30 and 33, rats were given SEB (0.5 mg/kg body weight) or PBS (control). Experimental groups were designated control, SEB, SEB-SDAP, and SEB-IgC. Lymphocyte populations were analyzed by immunohistochemistry. In lamina propria, SEB increased the cytotoxic lymphocyte populations of T-gammadelta cells (38%; P < 0.001) and natural killer cells (59%; P < 0.05) and the number of activated T lymphocytes (148%; P < 0.001). Both SDAP and IgC decreased the effects of SEB on these lymphocyte subsets (P < 0.05). In the epithelium, SEB induced a 117% increase in intraepithelial-activated lymphocytes that was reduced by SDAP supplementation (P < 0.01). The effects of plasma supplements on intestinal lymphocyte populations suggest that oral plasma proteins can modulate the degree of activation of diffuse GALT. PMID- 18287363 TI - Lycopene inhibits experimental metastasis of human hepatoma SK-Hep-1 cells in athymic nude mice. AB - Lycopene has been shown to inhibit tumor metastasis in vitro, but it is unclear whether lycopene is antimetastatic in vivo. Here, nude mice were orally supplemented 2 times per week for 12 wk with a low or high dose of lycopene [1 or 20 mg/kg body weight (BW)] or with beta-carotene (20 mg/kg BW). Two weeks after the beginning of supplementation, mice were injected once with human hepatoma SK Hep-1 cells via the tail vein. Plasma levels of matrix metalloproteinase (MMP)-2 and vascular endothelial growth factor (VEGF) increased gradually in tumor injected mice (tumor controls) following tumor injection but were markedly lowered by lycopene or beta-carotene supplementation. Ten weeks after tumor injection, mice were killed and tumor metastasis was found to be confined to the lungs. Compared with the tumor controls, high-lycopene supplementation lowered the mean number of tumors from 14 +/- 8 to 3 +/- 5 (P < 0.05) and decreased tumor cross-sectional areas by 62% (P < 0.05). High-lycopene supplementation also decreased the positive rate of proliferating cellular nuclear antigen (PCNA), the level of VEGF, and protein expressions of PCNA, MMP-9, and VEGF in lung tissues. However, high-lycopene increased the protein expression of nm23-H1 (an antimetastatic gene) by 133% (P < 0.001). For most variables measured, effects of lycopene were dose dependent and the effect of beta-carotene was between those of high-dose and low-dose lycopene. These results show that lycopene supplementation reduces experimental tumor metastasis in vivo and suggest that such an action is associated with attenuation of tumor invasion, proliferation, and angiogenesis. PMID- 18287364 TI - White button mushroom enhances maturation of bone marrow-derived dendritic cells and their antigen presenting function in mice. AB - Mushrooms have been shown to enhance immune response, which contributes to their antitumor property. White button mushrooms (Agaricus bisporus) (WBM) constitute 90% of the total mushrooms consumed in the United States; however, the health benefit of this strain in general is not well studied. Furthermore, little is known about WBM's immunologic effects. Dendritic cells (DC) are the most potent antigen presenting cells and play a pivotal role in immune response by linking innate and adaptive immune responses. In this study, we investigated the effect of in vitro supplementation with WBM on maturation of bone marrow-derived DC (BMDC) of C57BL mice. BMDC were differentiated in the presence of whole mushroom concentrate at 50, 100, or 200 mg/L. Results showed that mushroom supplementation dose dependently increased the expression of maturation markers CD40, CD80, CD86, and major histocompatibility complex-II. Consistent with the changes in the phenotypic markers, functional assay for DC maturation showed that mushroom supplementation decreased DC endocytosis and increased intracellular interleukin (IL)-12 levels. Furthermore, using a syngeneic T cell activation model, we found that WBM-supplemented DC from BALB/c mice presented ovalbumin antigen to T cells from DO11.10 mice more efficiently as demonstrated by increased T cell proliferation and IL-2 production. In conclusion, WBM promote DC maturation and enhance their antigen-presenting function. This effect may have potential in enhancing both innate and T cell-mediated immunity leading to a more efficient surveillance and defense mechanism against microbial invasion and tumor development. PMID- 18287365 TI - Lowering plasma homocysteine concentrations of older men and women with folate, vitamin B-12, and vitamin B-6 does not affect the proportion of (n-3) long chain polyunsaturated fatty acids in plasma phosphatidylcholine. AB - There is evidence to suggest that folate, homocysteine, or both affect the (n-3) long chain PUFA composition of tissues; however, this evidence is derived largely from experiments with animals and small observational studies in humans. Results from randomized controlled trials are needed. The objective of this study was to determine whether homocysteine lowering with a B vitamin supplement affects the proportion of (n-3) long-chain PUFA in plasma phosphatidylcholine. We conducted a double-blind, placebo-controlled, randomized clinical trial involving 253 participants, 65 y or older, with plasma homocysteine concentrations of at least 13 micromol/L. Participants in the vitamin group (n = 127) took a daily supplement containing 1000 microg folate, 500 microg vitamin B-12, and 10 mg vitamin B-6 for 2 y. The fatty acid composition of plasma phosphatidylcholine was measured at baseline and at 2 y. Plasma homocysteine concentrations during the course of the study were 4.4 micromol/L lower in the vitamin group than in the placebo group. The proportions of eicosapentaenoic, docosapentaenoic, and docosahexaenoic acids in plasma phosphatidylcholine did not differ between the vitamin and placebo groups at 2 y; the mean differences after adjusting for baseline values and sex were -0.03 (99% CI: -0.22, 0.16), 0.03 (99% CI: -0.03, 0.09), and -0.02 (99% CI: -0.27, 0.24) mol%, respectively. Lowering plasma homocysteine concentrations of older men and women with folate, vitamin B-12, and vitamin B-6 had no effect on the proportion of (n-3) long-chain PUFA in plasma phosphatidylcholine. PMID- 18287366 TI - A casein hydrolysate does not enhance gut endogenous protein flows compared with intact casein when fed to growing rats. AB - We studied the effect of dietary free peptides vs. peptides released naturally during digestion on gut endogenous nitrogen (N) flow (ENFL) and amino acid (AA) flow (EAAFL). Semisynthetic diets containing 110 g/kg diet of the same casein, intact (C) or hydrolyzed (HC), were formulated with TiO2 as a dietary marker. Sprague-Dawley rats were fed the diets hourly (0800-1500 h) for 10 min each hour for 7 d. Rats received unlabeled diets for 6 d and 15N-labeled diets on d 7, whereby they were killed and digesta sampled (6 observations per group) along the intestinal tract. EAAFL and ENFL were determined by 15N-isotope dilution (ID) for C or by ID or after centrifugation and ultrafiltration (UF) for HC. Ileal EAAFL and ENFL (ID) were not enhanced with diet HC compared with diet C. The AA compositions (g/16 g N) of ileal ENFL did not differ between rats fed HC and C except for Asp, Phe, Tyr, and Ser, for which contributions were relatively lower (P < 0.05) for rats fed C. Ileal EAAFL and ENFL (HC) were considerably lower (P < 0.05) with ID than with UF, but flows of Gly, Phe, and His were similar. There was no stimulatory effect of dietary peptides from HC on endogenous ileal protein flow compared with C, but the result is tentative given the high degree of dietary N recycled within endogenous protein and which could have occurred at a differential rate between rats fed diets C and HC. PMID- 18287367 TI - Dietary intake and major food sources of polyphenols in Finnish adults. AB - Phenolic acids, flavonoids, proanthocyanidins, and ellagitannins are polyphenols that may have beneficial effects on human health and provide protection against chronic diseases. To date, limited data exist on quantitative intake of polyphenols. The aims of this study were to estimate the quantitative intakes of polyphenols by using analyzed concentrations together with individual food consumption records and to determine major dietary sources. Analyzed concentrations of phenolic acids, anthocyanidins, and other flavonoids, proanthocyanidins, and ellagitannins (44 total polyphenol compounds) were entered into the national food composition database, Fineli. The absolute intakes of the polyphenols and the corresponding food sources were calculated on the basis of 48 h dietary recalls of 2007 Finnish adults. The mean total intake of polyphenols was 863 +/- 415 mg/d. Phenolic acids comprised the dominant group of polyphenols (75% of total intake) followed by proanthocyanidins (14%) and anthocyanidins and other flavonoids (10%). Due to their high consumption and high concentrations of phenolic acids, coffee and cereals were the main contributors to total polyphenol intake. Berries and berry products were the main source for anthocyanidins, ellagitannins, and proanthocyanidins, and fruits were the main source for flavonols, flavones, and flavanones. The results give additional support to the recommendations for a varied diet with fruits, berries, cereals, and vegetables. PMID- 18287368 TI - Validation of a food frequency questionnaire for assessing dietary soy isoflavone intake among midlife Chinese women in Hong Kong. AB - Substantial evidence suggests that soy isoflavones may protect against certain chronic diseases. This study aims to assess the reproducibility and validity of a 47-item semiquantitative soy FFQ (SFFQ) designed to measure the usual intake of soy isoflavones among 145 Chinese mid-life women in Hong Kong. Reproducibility of the SFFQ was assessed by the stability of dietary intake obtained at baseline (SFFQ0) and at 13-mo follow-up (SFFQ1). Relative validity was measured by comparing data collected from SFFQ1 with those derived from 23-d, 24-h dietary recalls (DR) collected during the same 1-y validation period. Isoflavone intake was calculated using analytical values from the Chinese University of Hong Kong Soy Isoflavone Database. A total of 3,217 DR were successfully obtained. The median (interquartile range) absolute intake estimated by the SFFQ1 was 0.91 ( 2.7 to 6.8) mg aglucon equivalents/d higher than the 6.3 (3.7-10.7) mg aglucon equivalents/d measured by the DR (P < 0.0057; Wilcoxon's Signed Rank test). Bland Altman analysis further demonstrated the presence of significant proportional bias between methods among Cantonese women with above-median intake (Spearman correlation coefficient; r = 0.44; P = 0.0005). Nonetheless, the intraclass and Pearson correlation coefficients, respectively, were 0.84 and 0.72 for non Cantonese and 0.65 and 0.50 for Cantonese, showing moderate to good levels of reproducibility and validity of the SFFQ (difference between 2 intraclass correlation coefficients, P = 0.09; difference between 2 Pearson r, P = 0.16). The unadjusted and BMI-adjusted correlations were of similar magnitude. The SFFQ is a reasonably valid instrument for assessing dietary soy isoflavone exposure in Hong Kong Chinese mid-life women. PMID- 18287369 TI - Vegetable but not fruit consumption reduces the risk of type 2 diabetes in Chinese women. AB - We examined associations between fruit and vegetable intake and the incidence of type 2 diabetes (T2D) in a population-based prospective study of 64,191 women with no history of T2D or other chronic diseases at study recruitment and with valid dietary information. Dietary intake was assessed by in-person interviews using a validated FFQ. During 297,755 person-years of follow-up, 1608 new cases of T2D were documented. We used a Cox regression model to evaluate the association of fruit and vegetable intake (g/d) with the risk of T2D. Quintiles of vegetable intake and T2D were inversely associated. The relative risk for T2D for the upper quintile relative to the lower quintile of vegetable intake was 0.72 (95%CI: 0.61-0.85; P < 0.01) in multivariate analysis. Individual vegetable groups were all inversely and significantly associated with the risk of T2D. Fruit intake was not associated with the incidence of diabetes in this population. Our data suggest that vegetable consumption may protect against the development of T2D. PMID- 18287371 TI - Development and validation of measure of household food insecurity in urban Costa Rica confirms proposed generic questionnaire. AB - Interest in household food insecurity (FI) within scientific and policy groups has motivated efforts to develop methods for measuring it. Questionnaires asking about FI experiences have been shown to be valid in the contexts in which they were created. The issue has arisen as to whether such questionnaires need be developed from the ground up or if a generic questionnaire can be adapted to a particular context. This study aimed to gain an in-depth understanding of household FI in urban Costa Rica, develop and validate a questionnaire for its measurement, and inform the choice between the 2 methods of development. The study was conducting using qualitative and quantitative methods provided in the Food and Nutrition Technical Assistance (FANTA) guidelines. In-depth interviews were conducted with 49 low-middle-income urban women using a semistructured interview guide. A 14-item FI questionnaire was developed based on results from these interviews. A field study was conducted in 213 households. The results show that the developed questionnaire provides valid measurement of household FI in urban Costa Rica and is simple and quick to apply in the household setting. FANTA developed a guide during the period that this research was completed that provides a generic questionnaire that can be adapted for use in various countries, rather than building the questionnaire from the ground up. This study provides evidence that careful attention to the procedures in this guide will likely yield a questionnaire suitable for assessing household FI in middle-income countries. PMID- 18287370 TI - Zinc, gravida, infection, and iron, but not vitamin B-12 or folate status, predict hemoglobin during pregnancy in Southern Ethiopia. AB - The etiology of anemia during pregnancy in rural Southern Ethiopia is uncertain. Intakes of animal-source foods are low and infections and bacterial overgrowth probably coexist. We therefore measured the dietary intakes of a convenience sample of Sidama women in late pregnancy who consumed either maize (n = 68) or fermented enset (Enset ventricosum) (n = 31) as their major energy source. Blood samples were analyzed for a complete blood count, vitamin B-12 and folate status, plasma ferritin, retinol, zinc, albumin, and C-reactive protein (CRP). The role of infection and gravida was also examined. Dietary intakes were calculated from 1-d weighed records. No cellular animal products were consumed. Of the women, 29% had anemia, 13% had iron deficiency anemia, 33% had depleted iron stores, and 74 and 27% had low plasma zinc and retinol, respectively. Only 2% had low plasma folate (< 6.8 nmol/L) and 23% had low plasma vitamin B-12 (< 150 pmol/L), even though 62% had elevated plasma methylmalonic acid (MMA) (> 271 nmol/L). None had elevated plasma cystathionine or total homocysteine (tHcys). Women with enset based diets had higher (P = 0.052) plasma vitamin B-12 concentration and lower (P < 0.05) cell volume, plasma cystathionine, and retinol than women consuming maize based diets, but mean hemoglobin, plasma ferritin, MMA, tHcys, and folate did not differ. Plasma zinc, followed by CRP (< or = 5 mg/L), gravida (< or = 4), and plasma ferritin (> or = 12 microg/L) status were major positive predictors of hemoglobin. Despite some early functional vitamin B-12 deficiency, there was no macrocytic anemia. Consumption of fermented enset may have increased vitamin B-12 levels in diet and plasma. PMID- 18287372 TI - An energy-dense complementary food is associated with a modest increase in weight gain when compared with a fortified porridge in Malawian children aged 6-18 months. AB - Poor complementary feeding practices are associated with stunting and growth faltering throughout the developing world. The objective was to compare the effect of using peanut-/soy-based fortified spread (FS) and corn porridge fortified with fish powder (FP) as complementary foods on growth in rural Malawian children. A total of 240 children were enrolled at the age of 6 mo and randomized to receive FS or FP. Both complementary foods provided 836 kJ/d from 6 to 9 mo of age and 1254 kJ/d from 9 to 18 mo of age. Children were followed monthly for anthropometry and fortnightly for the symptoms of fever, cough, or diarrhea until they were 18 mo old. Zn and Se status were assessed at 6 and 12 mo. The primary outcomes were the rates of weight and length gain from 6-12 mo and from 12-18 mo. Children who received FS gained 110 g more (95% CI 220 to 10) from 6-12 mo of age than children receiving FP. Weight gain did not differ between children receiving FS and FP between 12 and 18 mo of age, nor did statural growth from 6 to 12 mo or 12 to 18 mo. A total of 23% of all children were Zn deficient at 6 mo of age and this increased to 37% at 12 mo of age. Neither FS nor FP was associated with significantly improved Zn status. FS was associated with better weight gain from 6-12 mo of age and may be useful in conjunction with additional interventions to improve infant growth in the developing world. PMID- 18287373 TI - Breast-feeding patterns, time to initiation, and mortality risk among newborns in southern Nepal. AB - Initiation of breast-feeding within 1 h after birth has been associated with reduced neonatal mortality in a rural Ghanaian population. In South Asia, however, breast-feeding patterns and low birth weight rates differ and this relationship has not been quantified. Data were collected during a community based randomized trial of the impact of topical chlorhexidine antisepsis interventions on neonatal mortality and morbidity in southern Nepal. In-home visits were conducted on d 1-4, 6, 8, 10, 12, 14, 21, and 28 to collect longitudinal information on timing of initiation and pattern of breast-feeding. Multivariable regression modeling was used to estimate the association between death and breast-feeding initiation time. Analysis was based on 22,838 breast-fed newborns surviving to 48 h. Within 1 h of birth, 3.4% of infants were breast-fed and 56.6% were breast-fed within 24 h of birth. Partially breast-fed infants (72.6%) were at higher mortality risk [relative risk (RR) = 1.77; 95% CI = 1.32 2.39] than those exclusively breast-fed. There was a trend (P = 0.03) toward higher mortality with increasing delay in breast-feeding initiation. Mortality was higher among late (> or = 24 h) compared with early (< 24 h) initiators (RR = 1.41; 95% CI = 1.08-1.86) after adjustment for low birth weight, preterm birth, and other covariates. Improvements in breast-feeding practices in this setting may reduce neonatal mortality substantially. Approximately 7.7 and 19.1% of all neonatal deaths may be avoided with universal initiation of breast-feeding within the first day or hour of life, respectively. Community-based breast-feeding promotion programs should remain a priority, with renewed emphasis on early initiation in addition to exclusiveness and duration of breast-feeding. PMID- 18287374 TI - Food insecurity is associated with nutrient inadequacies among Canadian adults and adolescents. AB - Household food insecurity constrains food selection, but whether the dietary compromises associated with this problem heighten the risk of nutrient inadequacies is unclear. The objectives of this study were to examine the relationship between household food security status and adults' and children's dietary intakes and to estimate the prevalence of nutrient inadequacies among adults and children, differentiating by household food security status. We analyzed 24-h recall and household food security data for persons aged 1-70 y from the 2004 Canadian Community Health Survey (cycle 2.2). The relationship between adults' and children's nutrient and food intakes and household food security status was assessed using regression analysis. Estimates of the prevalence of inadequate nutrient intakes by food security status and age/sex group were calculated using probability assessment methods. Poorer dietary intakes were observed among adolescents and adults in food-insecure households and many of the differences by food security status persisted after accounting for potential confounders in multivariate analyses. Higher estimated prevalences of nutrient inadequacy were apparent among adolescents and adults in food insecure households, with the differences most marked for protein, vitamin A, thiamin, riboflavin, vitamin B-6, folate, vitamin B-12, magnesium, phosphorus, and zinc. Among children, few differences in dietary intakes by household food security status were apparent and there was little indication of nutrient inadequacy. This study indicates that for adults and, to some degree, adolescents, food insecurity is associated with inadequate nutrient intakes. These findings highlight the need for concerted public policy responses to ameliorate household food insecurity. PMID- 18287375 TI - The influence and benefits of controlling for inflammation on plasma ferritin and hemoglobin responses following a multi-micronutrient supplement in apparently healthy, HIV+ Kenyan adults. AB - Hemoglobin and ferritin are important biomarkers of iron status but are both altered by inflammation. We used the inflammation biomarkers C-reactive protein (CRP) and alpha1-acid glycoprotein (AGP) to adjust hemoglobin and ferritin concentrations to clarify interpretation of iron status. Apparently healthy adults who tested positive twice for HIV but who had not reached stage IV or clinical AIDS were randomly allocated to receive a food supplement (n = 17 and 21) or the food plus a micronutrient capsule (MN; 10 men and 34 women, respectively) containing 30 mg iron/d. Hemoglobin, ferritin, CRP, and AGP concentrations were measured at baseline and 3 mo and subjects were divided into 4 groups (reference, no inflammation; incubating, raised CRP; early convalescence, raised AGP and CRP; and late convalescence, raised AGP). Correction factors (the ratios of the median for the reference group over each inflammatory group) improved the consistency of the ferritin but not the hemoglobin results. After correction, ferritin (but not hemoglobin) increased in both men (48 microg/L; P = 0.02) and women (12 microg/L; P = 0.04) who received MN but not in the food-only group. However, hemoglobin did improve in subjects who showed no inflammation both at baseline and mo 3 (P = 0.019), but ferritin did not increase in this group. In conclusion, ferritin concentrations were more closely linked to current inflammation than hemoglobin; hence, correction by inflammation biomarkers improved data consistency. However, low hemoglobin concentrations were the consequence of long-term chronic inflammation and improvements in response to MN supplements were only detected in subjects with no inflammation. PMID- 18287376 TI - Neighborhood socioeconomic deprivation and minority composition are associated with better potential spatial access to the ground-truthed food environment in a large rural area. AB - Little is known about spatial inequalities and potential access to the food environment in rural areas. In this study, we assessed the food environment in a 6-county rural region of Texas (11,567 km2) through ground-truthed methods that included direct observation and on-site Global Positioning System technology to examine the relationship between neighborhood inequalities (e.g., socioeconomic deprivation and minority composition) and network distance from all 101 rural neighborhoods to the nearest food store (FS). Neighborhood deprivation was determined from socioeconomic characteristics using 2000 census block group (CBG) data. Network distances were calculated from the population-weighted center of each CBG to the nearest supermarket, grocery, convenience, and discount store. Multiple regression models examined associations among deprivation, minority composition, population density, and network distance to the nearest FS. The median distance to the nearest supermarket was 14.9 km one way (range 0.12 to 54.0 km). The distance decreased with increasing deprivation, minority composition, and population density. The worst deprived neighborhoods with the greatest minority composition had better potential spatial access to the nearest FS. For >20% of all rural residents, their neighborhoods were at least 17.7 km from the nearest supermarket or full-line grocery or 7.6 km from the nearest convenience store. This makes food shopping a challenge, especially in rural areas that lack public transportation and where many have no vehicular access. Knowledge of potential access to the food environment is essential for combining environmental approaches and health interventions so that families, especially those in rural areas, can make healthier food choices. PMID- 18287377 TI - From efficacy to public health impact: a call for research on program delivery and utilization in nutrition. AB - Despite the high potential of nutrition interventions to save lives and to improve the quality of life of those who survive, programs are not being implemented effectively or at scale in most regions of the world. Research on delivery and utilization is urgently needed to fully unleash the potential of nutrition programs. The symposium sought to explore knowledge on determinants of delivery and utilization, demonstrate approaches for studying them, and highlight future research needs. PMID- 18287378 TI - A results framework serves both program design and delivery science. AB - Health programmers and researchers must collaborate despite different mandates and technical languages. A results framework is a simple model that both disciplines can use to understand complexity, clarify assumptions and hypotheses, design programs, and ask questions to inform action research. Typically, a health program's results framework has 3 tiers and 6 boxes: a base of 4 health service intermediate results (access, quality, demand, and environment), which lead to a midlevel strategic objective (use of life-saving intervention), which leads to the goal (improved health). A situation analysis directly informs intervention selection; more difficult is selecting strategies to deliver the interventions, especially in settings of health system weakness. We propose menus and submenus of strategies to achieve each intermediate result, illustrate the use of the results framework in a program design and in clarifying research questions, and begin to propose a research agenda for "delivery scientists" responsible for recommending optimal investments to maximize use of interventions by those who need them most. PMID- 18287379 TI - Assessing supervisory and motivational factors in the context of a program evaluation in rural Haiti. AB - Staff supervisory and motivational factors were assessed in the context of an ongoing program evaluation in Haiti comparing 2 models of targeting an integrated health and nutrition program. The study objectives were to 1) understand and improve supervisory and motivational factors influencing program implementation and 2) compare these factors between the 2 program models being evaluated. Qualitative methods (focus group discussions and semistructured interviews) were used to understand factors related to supervision and motivation. Quantitative measures of supervisory and motivational factors were designed, and factor analysis was used, to develop summary scales of motivational factors and supervision. T-tests were used to compare mean scores on the scales between the 2 program models. Results from the qualitative research were discussed with program management and staff to help develop solutions to implementation bottlenecks. Staff at all levels of the program seemed motivated and generally well supervised. Constraints to motivation included perceived inadequacy of wages (before changes were made to salaries), heavy workloads, and logistical constraints. We found no salient differences between the 2 program models that could contribute to differential implementation or differences in impact. This lack of salient differences between the program models suggested that supervisory and motivational factors were unlikely to contribute to differences in impact. Assessing supervisory and motivational factors was feasible and desirable in the context of this evaluation and deepened understanding of the program context and constraints to implementation. PMID- 18287380 TI - Improving enrollment and utilization of the Oportunidades program in Mexico could increase its effectiveness. AB - Oportunidades, Mexico's most important antipoverty program, currently with 5 million enrolled households in all regions of the country, has been shown to significantly contribute to improving the nutrition, health, and education of the poor. Because the program has used different enrollment strategies in rural and urban areas and has both obligatory (e.g., health and nutrition education) and nonobligatory components (e.g., nutrition supplements for children younger than 2 y of age), it provides an excellent opportunity to study program enrollment and utilization of different program components. In urban areas enrollment was more complex, and hence enrollment was much lower then in rural areas where the process was quasiautomatic, and nearly all eligible households enrolled. Enrollment in urban areas was not associated with having a child younger than 2 y of age. Utilization was notably higher with the obligatory than with the nonobligatory program components, illustrated by the inadequate consumption of the nutrition program's supplement as compared with near-universal compliance with well-baby visits. Innovative approaches, some of which are currently being tested, are needed to further increase the program's impact. PMID- 18287381 TI - Differential community response to introduction of zinc for childhood diarrhea and combination therapy for malaria in southern Mali. AB - Developing effective, affordable, and sustainable delivery strategies for the isolated low-income populations that stand to gain the most from micronutrient interventions has proven difficult. We discuss our experience with implementation of zinc as treatment for diarrhea in children less than 5 y of age over the course of 3 operational research studies in rural Sikasso Region, Mali, West Africa. The initial formative research study highlighted how malaria affects perceptions of diarrhea and its causes and that malaria and diarrhea are not necessarily viewed as distinct conditions. The second-phase pilot introduction demonstrated that, in introducing zinc treatment in malaria-endemic regions, it is especially important that both community- and facility-level providers be trained to manage sick children presenting with multiple symptoms. The third phase study on large-scale implementation detected that the experience with implementation of new treatments for malaria is distinct from that of diarrhea. To some extent zinc treatment is the solution to a problem that communities may not recognize at all. Interventions to improve case management of sick children must be integrated across diseases and nutritional problems at both the facility and community levels. Operational research can identify points where integration should occur and how it should be carried out. Programs targeting single diseases or single nutritional problems can have a variety of deleterious effects on health systems, no matter how well they are planned. PMID- 18287382 TI - Improving results for nutrition: a commentary on an agenda and the need for implementation research. AB - Research and implementation often exist in separate worlds. To improve results for nutrition, the nutrition research community needs to go beyond "what" works to understand "how" it works. If they do not, nutrition research risks becoming irrelevant to the needs of those who actually make policies and implement programs. Researchers must prioritize research on effectiveness of policies and programs. They should incorporate knowledge and tools of social sciences, including economics, sociology, political science, and management into their work. They should pay greater attention to environmental and institutional variables and understand change strategies, knowledge utilization, and policy processes. Fundamentally, research on implementation should use a systematic approach to produce generalizable evidence and conceptual models, tools, and methods that are communicated effectively to policymakers and programmers. Nutrition researchers need not expand far beyond their disciplinary comfort zone to do this, but they do need to build bridges with other fields to have greater success in addressing nutritional challenges. PMID- 18287383 TI - Nav1.9, G-proteins, and nociceptors. PMID- 18287384 TI - Gerbils can tune in. PMID- 18287385 TI - GABAA receptor subunit specificity: a tonic for the excited brain. PMID- 18287386 TI - Evidence of a physiological role for use-dependent inactivation of NaV1.8 sodium channels. PMID- 18287387 TI - Cancer statistics, 2008. AB - Each year, the American Cancer Society estimates the number of new cancer cases and deaths expected in the United States in the current year and compiles the most recent data on cancer incidence, mortality, and survival based on incidence data from the National Cancer Institute, Centers for Disease Control and Prevention, and the North American Association of Central Cancer Registries and mortality data from the National Center for Health Statistics. Incidence and death rates are age-standardized to the 2000 US standard million population. A total of 1,437,180 new cancer cases and 565,650 deaths from cancer are projected to occur in the United States in 2008. Notable trends in cancer incidence and mortality include stabilization of incidence rates for all cancer sites combined in men from 1995 through 2004 and in women from 1999 through 2004 and a continued decrease in the cancer death rate since 1990 in men and since 1991 in women. Overall cancer death rates in 2004 compared with 1990 in men and 1991 in women decreased by 18.4% and 10.5%, respectively, resulting in the avoidance of over a half million deaths from cancer during this time interval. This report also examines cancer incidence, mortality, and survival by site, sex, race/ethnicity, education, geographic area, and calendar year, as well as the proportionate contribution of selected sites to the overall trends. Although much progress has been made in reducing mortality rates, stabilizing incidence rates, and improving survival, cancer still accounts for more deaths than heart disease in persons under age 85 years. Further progress can be accelerated by supporting new discoveries and by applying existing cancer control knowledge across all segments of the population. PMID- 18287388 TI - Publication ethics and scientific misconduct: the role of authors. PMID- 18287389 TI - A case of anterior open bite with severely narrowed maxillary dental arch and hypertrophic palatine tonsils. AB - This case report describes an adolescent patient with an open bite and severely narrowed maxillary dentition and hypertrophic palatine tonsils, treated efficiently with rapid maxillary expansion (RME) and subsequent orthodontic tooth alignment using fixed appliances. The treatment demonstrates that RME can be effective for the correction of a severely narrowed maxillary arch, as well as, in this case, the correction of an anterior open bite in an adolescent patient where no substantial vertical skeletal discrepancy existed. PMID- 18287390 TI - Beware the solitary maxillary median central incisor. AB - The incidence of a solitary maxillary median central incisor (SMMCI) tooth in the general population is low, in either the primary or secondary dentition. The most common cause of a missing maxillary central incisor is trauma, or more rarely hypodontia. However, SMMCI is also a recognized genetic anomaly and affected individuals can be carriers for a potentially more serious condition affecting midline development of the brain and face, holoprosencephaly (HPE). The presence of an SMMCI of unknown aetiology is therefore considered a risk factor for HPE, even in the absence of any other clinical signs. The orthodontist may be responsible for diagnosing cases of SMMCI with no obvious cause, and in these subjects due consideration should be given to referral for the appropriate genetic testing and counselling. PMID- 18287391 TI - The unerupted maxillary second molar, due to an overlying and malformed upper third molar: treatment and follow-up. AB - This retrospective study presents the treatment and follow-up of 20 young patients with 23 impacted upper second molars, due to overlying, impacted upper third molars. The third molars were removed surgically under local anaesthesia. After removal of these palatally obstructing teeth, radiographic and clinical follow-up was performed. The purpose of this study was to evaluate the eruption progress of the upper second molars after surgery. Radiological and/or clinical follow-up showed complete eruption of 19 (83%) of the upper second molars. For those cases treated before the age of 12 years and 4 months (the mean eruption age), all the upper second molars erupted completely. For those cases where surgical removal was undertaken after the mean eruption age, four (17%) of the upper second molars did not completely erupt. It was concluded that early treatment of impacted upper second molars, due to overlying third molars, may lead to more rapid eruption. Further prospective research is necessary to develop guidelines for the removal of palatally obstructing third molars to avoid eruption problems. PMID- 18287392 TI - A randomized control clinical trial investigating orthodontic bond failure rates when using Orthosolo universal bond enhancer compared to a conventional bonding primer. AB - OBJECTIVE: This study assessed the in vivo bond failure rates of orthodontic brackets bonded using Orthosolo universal bond enhancer and compared it with the conventional bonding primer, Transbond XT. DESIGN: This was a single centre randomized controlled clinical study. SETTING: Department of Child Dental Health, Bristol Dental Hospital, Bristol, UK. MATERIALS AND METHODS: Thirty-three consecutive patients undergoing fixed orthodontic appliance therapy were included in this study. Using a split-mouth design, diagonally opposite quadrants were randomly allocated a primer, either Orthosolo universal bond enhancer (Ormco, Orange, CA, USA) or Transbond XT primer (3M Unitek, Monrovia, CA, USA). A total of 555 teeth were bonded using a conventional acid-etch technique. 277 received Orthosolo as their primer and 278 received the Transbond XT primer. Bond failures and their positions were recorded at six months. RESULTS: There was an overall bond failure rate of 1.26%. Four brackets failed in the Orthosolo group (0.72%) and three failed in the Transbond XT group (0.54%). CONCLUSION: There was no clinical or statistically significant difference in the in vivo bond failure rates between orthodontic brackets bonded using either Orthosolo universal bond enhancer or the conventional Transbond XT primer. PMID- 18287393 TI - Space conditions and prevalence of anterior spacing and crowding among nine-year old schoolchildren. AB - OBJECTIVE: To give detailed insight into the space conditions and prevalence of spacing and crowding in schoolchildren at nine years of age. The reliability and validity of screening methods was also assessed. DESIGN: Epidemiological survey. SETTING: South-western part of Germany (Rhein-Neckar-Kreis); elementary schools in a region with a low orthodontic care rate. SAMPLE AND METHODS: 494 nine-year old German schoolchildren (237 males, 257 females, median age 9 years) were examined orthodontically in cooperation with the local community dentistry service. A special measurement ruler was developed to enhance validity and reliability of space measurements on location in the schools. RESULTS AND CONCLUSIONS: Concerning space conditions in the incisor segments: in the maxilla the variety encountered comprised both space excess and moderate to severe crowding. This was considerably larger in the maxilla than in the mandible. Severe crowding (> 5 mm) was found more often in the maxilla than in the mandible and affected around 2-3% of the maxillary dentitions. Also severe contact point displacements (IOTN Grade 4) were mainly restricted to the maxilla (prevalence approximately 3%). In the canine-premolar segments, the arch segments in males were in general around 0.5 mm larger than among the females, also within each dental stage. In general, the maxilla was more often affected by posterior crowding than the mandible. Anterior crowding seemed to be more prevalent than posterior crowding. With respect to the screening methods used in the present study, reliable and valid measurements were also found to be possible in schools. This may open up further opportunities for orthodontic screening by community dentistry services or similar organizations. PMID- 18287394 TI - Effect of sandblasting on the retention of orthodontic brackets: a controlled clinical trial. AB - OBJECTIVE: To study the effect of chairside sandblasting of the bases upon the retention of mesh backed orthodontic brackets. DESIGN: Prospective controlled clinical trial. METHODS: Brackets were bonded to 60 successive patients who were treated at an orthodontic practice in Amman, Jordan. Using a crossover system of allocation, quadrants were bonded using either sandblasted or non-sandblasted brackets respectively. Rely-A-Bond adhesive was used throughout. Bond failures were monitored over one year. RESULTS: 1112 brackets were assessed. The overall failure rate was 4.0% and the failure rates for non-sandblasted and sandblasted brackets were 4.7 and 3.4% respectively. The odds ratio for at least one bracket failure within the control quadrant compared with the experimental quadrant was 0.50 (95% CI 0.185 to 1.238), which was not statistically significant. CONCLUSION: Sandblasting did not significantly improve the retention of mesh based orthodontic brackets in this study. PMID- 18287397 TI - Choosing a pre-adjusted orthodontic appliance prescription for anterior teeth. PMID- 18287398 TI - Increased renal renin content in mice lacking the Na+/H+ exchanger NHE2. AB - Macula densa (MD) cells express the Na(+)/H(+) exchanger (NHE) isoform NHE2 at the apical membrane, which may play an important role in tubular salt sensing through the regulation of cell volume and intracellular pH. These studies aimed to determine whether NHE2 participates in the MD control of renin synthesis. Renal renin content and activity and elements of the MD signaling pathway were analyzed using wild-type (NHE2(+/+)) and NHE2 knockout (NHE2(-/-)) mice. Immunofluorescence studies indicated that NHE2(-/-) mice lack NHE3 at the MD apical membrane, so the other apical NHE isoform has not compensated for the lack of NHE2. Importantly, the number of renin-expressing cells in the afferent arteriole in NHE2(-/-) mice was increased approximately 2.5-fold using renin immunohistochemistry. Western blotting confirmed approximately 20% higher renal cortical renin content in NHE2(-/-) mice compared with wild type. No-salt diet for 1 wk significantly increased renin content and activity in NHE2(+/+) mice, but the response was blunted in NHE2(-/-) mice. Renal tissue renin activity and plasma renin concentration were elevated three- and twofold, respectively, in NHE2(-/-) mice compared with wild type. NHE2(-/-) mice also exhibited a significantly increased renal cortical cyclooxygenase-2 (COX-2) and microsomal prostaglandin E synthase (mPGES) expression, indicating MD-specific mechanisms responsible for the increased renin content. Significant and chronic activation of ERK1/2 was observed in MD cells of NHE2(-/-) kidneys. Removal of salt or addition of NHE inhibitors to cultured mouse MD-derived (MMDD1) cells caused a time-dependent activation of ERK1/2. In conclusion, the NHE2 isoform appears to be important in the MD feedback control of renin secretion, and the signaling pathway likely involves MD cell shrinkage and activation of ERK1/2, COX-2, and mPGES, all well-established elements of the MD-PGE(2)-renin release pathway. PMID- 18287399 TI - Transcriptional control of terminal nephron differentiation. AB - Terminal differentiation of epithelial cells into more specialized cell types is a critical step in organogenesis. Throughout the process of terminal differentiation, epithelial progenitors acquire or upregulate expression of renal function genes and cease to proliferate, while expression of embryonic genes is repressed. This exquisite coordination of gene expression is accomplished by signaling networks and transcription factors which couple the external environment with the new functional demands of the cell. While there has been much progress in understanding the early steps involved in renal epithelial cell differentiation, a major gap remains in our knowledge of the factors that control the steps of terminal differentiation. A number of signaling molecules and transcription factors have been recently implicated in determining segmental nephron identity and functional differentiation. While some of these factors (the p53 gene family, hepatocyte nuclear factor-1beta) promote the terminal epithelial differentiation fate, others (Notch, Brn-1, IRX, KLF4, and Foxi1) tend to regulate differentiation of specific nephron segments and individual cell types. This review summarizes current knowledge related to these transcription factors and discusses how diverse cellular signals are integrated to generate a transcriptional output during the process of terminal differentiation. Since these transcriptional processes are accompanied by profound changes in nuclear chromatin structure involving the genes responsible for creating and maintaining the differentiated cell phenotype, future studies should focus on identifying the nature of these epigenetic events and factors, how they are regulated temporally and spatially, and the chromatin environment they eventually reside in. PMID- 18287400 TI - The impact of aging on kidney repair. AB - The process of normal aging affects organ homeostasis as well as responses to acute and chronic injury. In view of the rapid growth in the elderly population, it is increasingly important for us to develop a mechanistic understanding of how these age-dependent changes can impact the susceptibility and response of the kidney to injurious stimuli. In this overview, we focus on the current understanding of those mechanisms by reviewing how cellular changes in the aging kidney might lead to a diminished proliferative reserve, an increased tendency for apoptosis, alterations in growth factor profiles, and changes in potential progenitor and immune cell functions. A better understanding of these processes may help us to define new targets for studying kidney repair and could ultimately lead to new therapeutic strategies that are specifically tailored for treatment of the elderly population. PMID- 18287401 TI - Nicotine-activated descending facilitation on spinal NMDA-dependent reflex potentiation from pontine tegmentum in rats. AB - This study was conducted to investigate the possible neurotransmitter that activates the descending pathways coming from the dorsolateral pontine tegmentum (DPT) to modulate spinal pelvic-urethra reflex potentiation. External urethra sphincter electromyogram (EUSE) activity in response to test stimulation (TS, 1/30 Hz) and repetitive stimulation (RS, 1 Hz) on the pelvic afferent nerve of 63 anesthetized rats were recorded with or without microinjection of nicotinic cholinergic receptor (nAChR) agonists, ACh and nicotine, to the DPT. TS evoked a baseline reflex activity with a single action potential (1.00 +/- 0.00 spikes/stimulation, n = 40), whereas RS produced a long-lasting reflex potentiation (16.14 +/- 0.96 spikes/stimulation, n = 40) that was abolished by d 2-amino-5-phosphonovaleric acid (1.60 +/- 0.89 spikes/stimulation, n = 40) and was attenuated by 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo (F) quinoxaline (7.10 +/- 0.84 spikes/stimulation, n = 40). ACh and nicotine microinjections to DPT both produced facilitation on the RS-induced reflex potentiation (23.57 +/- 2.23 and 28.29 +/- 2.36 spikes/stimulation, P < 0.01, n = 10 and 20, respectively). Pretreatment of selective nicotinic receptor antagonist, chlorisondamine, reversed the facilitation on RS-induced reflex potentiation caused by nicotine (19.41 +/- 1.21 spikes/stimulation, P < 0.01, n = 10) Intrathecal WAY-100635 and spinal transection at the T(1) level both abolished the facilitation on reflex potentiation resulting from the DPT nicotine injection (12.86 +/- 3.13 and 15.57 +/- 1.72 spikes/stimulation, P < 0.01, n = 10 each). Our findings suggest that activation of nAChR at DPT may modulate N-methyl-d-aspartic acid-dependent reflex potentiation via descending serotonergic neurotransmission. This descending modulation may have physiological/pathological relevance in the neural controls of urethral closure. PMID- 18287402 TI - Preeclamptic sera induce nephrin shedding from podocytes through endothelin-1 release by endothelial glomerular cells. AB - In preeclampsia (PE), proteinuria has been associated with a reduced expression of nephrin by podocytes. In the present study, we investigated in vitro on human cultured podocytes the mechanism responsible for nephrin loss in PE. Sera from patients with PE did not directly downregulate the expression of nephrin. In contrast, conditioned medium obtained from glomerular endothelial cells incubated with PE sera induced loss of nephrin and synaptopodin, but not of podocin, from podocytes. Nephrin loss was related to a rapid shedding of the protein from the cell surface due to cleavage of its extracellular domain by proteases and to cytoskeleton redistribution. The absence of nephrin mRNA downregulation together with nephrin reexpression within 24 h confirm that the loss of nephrin was not related to a reduced synthesis. Studies with an endothelin-1 (ET-1) receptor antagonist that abrogated the loss of nephrin triggered by glomerular endothelial conditioned medium of PE sera indicated that ET-1 was the main effector of nephrin loss. Indeed, ET-1 was synthesized and released from glomerular endothelial cells when incubated with PE sera, and recombinant ET-1 triggered nephrin shedding from podocytes. Moreover, VEGF blockade induced ET-1 release from endothelial cells, and in turn the conditioned medium obtained triggered nephrin loss. In conclusion, the present study identifies a potential mechanism of nephrin loss in PE that may link endothelial injury with enhanced glomerular permeability. PMID- 18287403 TI - Role of angiotensin II in the enhancement of ammonia production and secretion by the proximal tubule in metabolic acidosis. AB - Acidosis and angiotensin II stimulate ammonia production and transport by the proximal tubule. We examined the modulatory effect of the type 1 angiotensin II receptor blocker losartan on the ability of metabolic acidosis to stimulate ammonia production and secretion by mouse S2 proximal tubule segments. Mice given NH(4)Cl for 7 days developed metabolic acidosis (low serum bicarbonate concentration) and increased urinary excretion of ammonia. S2 tubule segments from acidotic mice displayed higher rates of ammonia production and secretion compared with those from control mice. However, when losartan was coadministered in vivo with NH(4)Cl, both the acidosis-induced increase in urinary ammonia excretion and the adaptive increase in ammonia production and secretion of microperfused S2 segments were largely blocked. In renal cortical tissue, losartan blocked the acid-induced increase in brush-border membrane NHE3 expression but had no effect on the acid-induced upregulation of phosphate dependent glutaminase or phosphoenolpyruvate carboxykinase 1 in cortical homogenates. Addition of angiotensin II to the microperfusion solution enhanced ammonia secretion and production rates in tubules from NH(4)Cl-treated and control mice in a losartan-inhibitable manner. These results demonstrate that a 7 day acid challenge induces an adaptive increase in ammonia production and secretion by the proximal tubule and suggest that during metabolic acidosis, angiotensin II signaling is necessary for adaptive enhancements of ammonia excretion by the kidney and ammonia production and secretion by S2 proximal tubule segments, as mediated, in part, by angiotensin receptor-dependent enhancement of NHE3 expression. PMID- 18287405 TI - Robert L. Metzenberg, June 11, 1930-July 15, 2007: geneticist extraordinaire and "model human". PMID- 18287406 TI - Recessive black is allelic to the yellow plumage locus in Japanese quail and associated with a frameshift deletion in the ASIP gene. AB - The recessive black plumage mutation in the Japanese quail (Coturnix japonica) is controlled by an autosomal recessive gene (rb) and displays a blackish-brown phenotype in the recessive homozygous state (rb/rb). A similar black coat color phenotype in nonagouti mice is caused by an autosomal recessive mutation at the agouti locus. An allelism test showed that wild type and mutations for yellow, fawn-2, and recessive black in Japanese quail were multiple alleles (*N, *Y, *F2, and *RB) at the same locus Y and that the dominance relationship was Y*F2 > Y*Y > Y*N > Y*RB. A deletion of 8 bases was found in the ASIP gene in the Y*RB allele, causing a frameshift that changed the last six amino acids, including a cysteine residue, and removed the normal stop codon. Since the cysteine residues at the C terminus are important for disulphide bond formation and tertiary structure of the agouti signaling protein, the deletion is expected to cause a dysfunction of ASIP as an antagonist of alpha-MSH in the Y*RB allele. This is the first evidence that the ASIP gene, known to be involved in coat color variation in mammals, is functional and has a similar effect on plumage color in birds. PMID- 18287407 TI - Characterization of Japanese quail yellow as a genomic deletion upstream of the avian homolog of the mammalian ASIP (agouti) gene. AB - ASIP is an important pigmentation gene responsible for dorsoventral and hair cycle-specific melanin-based color patterning in mammals. We report some of the first evidence that the avian ASIP gene has a role in pigmentation. We have characterized the genetic basis of the homozygous lethal Japanese quail yellow mutation as a >90-kb deletion upstream of ASIP. This deletion encompasses almost the entire coding sequence of two upstream loci, RALY and EIF2B, and places ASIP expression under control of the RALY promoter, leading to the presence of a novel transcript. ASIP mRNA expression was upregulated in many tissues in yellow compared to wild type but was not universal, and consistent differences were not observed among skins of yellow and wild-type quail. In a microarray analysis on developing feather buds, the locus with the largest downregulation in yellow quail was SLC24A5, implying that it is regulated by ASIP. Finally, we document the presence of ventral skin-specific isoforms of ASIP mRNA in both wild-type quails and chickens. Overall, there are remarkable similarities between yellow in quail and lethal yellow in mouse, which involve a deletion in a similar genomic position. The presence of ventral-specific ASIP expression in birds shows that this feature is conserved across vertebrates. PMID- 18287408 TI - Completely distinguishing individual A-genome chromosomes and their karyotyping analysis by multiple bacterial artificial chromosome - fluorescence in situ hybridization. AB - Twenty bacterial artificial chromosome (BAC) clones that could produce bright signals and no or very low fluorescence in situ hybridization (FISH) background were identified from Gossypium arboreum cv. JLZM, and G. hirsutum accession (acc.) TM-1 and 0-613-2R. Combining with 45S and 5S rDNA, a 22-probe cocktail that could identify all 13 G. arboreum chromosomes simultaneously was developed. According to their homology with tetraploid cotton, the G. arboreum chromosomes were designated as A1-A13, and a standard karyotype analysis of G. arboreum was presented. These results demonstrated an application for multiple BAC-FISH in cotton cytogenetic studies and a technique to overcome the problem of simultaneous chromosome recognition in mitotic cotton cells. PMID- 18287409 TI - Anatomic features of metatarsal heads that simulate erosive disease: cadaveric study using CT, radiography, and dissection with special emphasis on cross sectional characterization of osseous anatomy. AB - OBJECTIVE: The purpose of our study was to characterize and describe normal osseous contours of the metatarsal heads that may simulate erosive changes. MATERIALS AND METHODS: CT of the metatarsal heads was performed in six human cadaveric feet, and 3D reformatted images were analyzed. Subsequently, five feet were sectioned in planes corresponding to those at imaging for anatomic correlation, and one foot was anatomically dissected for correlation. The normal anatomic osseous contours of the metatarsal heads were analyzed and described. RESULTS: Normal osseous anatomy of the greater (first) and lesser (second through fifth) metatarsal heads was identified. The normal osseous landmarks that simulate erosive changes were identified. Variations in the normal anatomic osseous contours, including those related to the medial and lateral condyles, were present in all specimens. The lateral condyle of the first metatarsal head was found to be more prominent than the medial condyle. In addition, anatomic variations in the normal osseous concavities of the lateral and medial aspects of each metatarsal head were measured. An intersesamoidal ridge, present on the plantar surface of the first metatarsal head, was identified in all specimens. CONCLUSION: The normal anatomic contours of the metatarsal head are a potential major source for diagnostic error when viewing sectional CT and MR images in patients with suspected erosive arthritis. These normal variations, although common and varied, produce characteristic findings that can be differentiated from bone erosions. PMID- 18287410 TI - Normal anatomy and strains of the deep musculotendinous junction of the proximal rectus femoris: MRI features. AB - OBJECTIVE: The MRI features of the proximal rectus femoris musculotendinous junction have scarcely been described in the literature. The purpose of our study, based on a review of 50 asymptomatic and 20 symptomatic MRI studies, was to define the normal MRI anatomy and MRI features of intrasubstance injury of deep musculotendinous tears of the proximal rectus femoris. CONCLUSION: Axial and coronal MR images are optimal for visualizing the direct and indirect heads, the conjoined tendon, and the deep musculotendinous junction of the proximal rectus femoris. Tears of the deep musculotendinous junction are longitudinal, involving a long segment of the muscle. MRI features include a "bull's-eye" sign, longitudinal scar, retraction, pseudocyst, and hematoma. PMID- 18287411 TI - Combined percutaneous radiofrequency ablation and ethanol injection for hepatocellular carcinoma in high-risk locations. AB - OBJECTIVE: The purpose of this study was to investigate whether combining percutaneous ethanol injection (PEI) with radiofrequency ablation in the management of hepatocellular carcinoma (HCC) in high-risk locations improves treatment outcomes. SUBJECTS AND METHODS: We compared the outcome of management of high-risk tumors with PEI and radiofrequency ablation (n = 50) or radiofrequency ablation alone (n = 114) with the outcome of radiofrequency ablation of non-high-risk tumors (n = 44). We also compared the survival rates of patients with and those without high-risk HCC. PEI was performed into the part of the tumor closest to a blood vessel or vital structure before radiofrequency ablation. RESULTS: The study included 142 patients with 208 HCCs managed with radiofrequency ablation. Despite larger tumor sizes (2.8 +/- 1 cm vs 1.9 +/- 0.7 cm vs 2.5 +/- 0.1 cm for the high-risk radiofrequency plus PEI, non-high-risk radiofrequency, and high-risk radiofrequency groups, respectively; p < 0.001), the primary effectiveness rate of high-risk radiofrequency ablation and PEI (92%) was similar to that of non-high-risk radiofrequency ablation (96%). The primary effectiveness rate of high-risk radiofrequency ablation and PEI was slightly higher (p = 0.1) than that of high-risk radiofrequency ablation (85%). The local tumor progression rates (21% vs 33% vs 24% at 18 months) of the three respective groups were not statistically different (p = 0.91). Patients with and those without high-risk tumors had equal survival rates (p = 0.42) after 12 (87% vs 100%) and 24 (77% vs 80%) months of follow-up. Independent predictors of primary effectiveness were a tumor size of 3 cm or less (p = 0.01) and distinct tumor borders (p = 0.009). Indistinct borders (p = 0.033) and non-treatment-naive status of HCC (p = 0.002) were associated with higher local tumor progression rates. The only predictor of survival was complete ablation of all index tumors (p = 0.001). CONCLUSION: The combination of radiofrequency ablation and PEI in the management of HCC in high-risk locations has a slightly higher primary effectiveness rate than does radiofrequency ablation alone. A randomized controlled study is warranted. PMID- 18287412 TI - Intraoperative loss of core biopsy clips: clinical implications. AB - OBJECTIVE: The purpose of this study was to report the occurrence of intraoperative loss of metallic marking clips placed during image-guided biopsy and to hypothesize the likely mechanism of this clinical problem. MATERIALS AND METHODS: From January 2003 through December 2004, patients presenting for preoperative mammographic localization and operative excision of biopsy site marking clips were identified. Age, method of image-guided biopsy, number of excised specimens, and tissue diagnosis were determined. Specimen radiographs were used to identify cases of suspected intraoperative clip loss. Clips absent on specimen radiographs and postoperative mammograms were defined as lost intraoperatively. Biopsy site marking clips, surgical clips, and suction device apertures were measured. RESULTS: In 78 surgical procedures performed during the study period, three (3.8%) of the patients experienced clip loss. Specimen radiographs confirmed the absence of clips in all submitted tissues. A median of four (range, three to five) separate biopsy specimens were excised among these three cases. A healing biopsy site from the stereotactic biopsy preceding the clip placement procedure was confirmed in all cases. Absence of the metallic clip was confirmed on postoperative mammograms. The apertures of two types of suction device were four and two times those of the biopsy clips. CONCLUSION: Intraoperative loss of metallic clips placed at the conclusion of image-guided breast biopsy is unusual but can occur during subsequent surgical excision. Repeated inability to locate the clip on specimen radiographs after accurate preoperative localization should raise the suspicion that the target clip has been lost, not missed, during surgery, likely because of inadvertent removal of the clip with the suction device. PMID- 18287413 TI - Differentiating cirrhosis and chronic hepatosplenic schistosomiasis using MRI. AB - OBJECTIVE: The objective of our study was to identify which imaging features may be used to differentiate between cirrhosis and chronic hepatosplenic schistosomiasis and to assess image interpretation agreement for MRI findings. MATERIALS AND METHODS: Retrospective review of 27 patients with alcoholic or virus-induced cirrhosis and 24 patients with chronic hepatosplenic schistosomiasis who underwent MRI (1.5 T) of the abdomen was performed. Images were interpreted independently by two radiologists evaluating the following MRI features: hepatic fissure widening, irregularity of hepatic contours, periportal fibrosis, hepatic parenchyma heterogeneity, and splenic siderotic nodules. Left, right, and caudate hepatic lobe measurements were obtained, and the splenic index was measured. The Fisher's exact test, chi-square test, and Student's t test were used to compare both groups, and regression analysis was performed. Observer agreement was measured using kappa and intraclass correlation tests. RESULTS: Periportal fibrosis, heterogeneity of hepatic parenchyma, and splenic siderotic nodules were more frequent in the group with schistosomiasis (p < 0.05), with periportal fibrosis showing the largest difference in presence and distribution (peripheral greater than central). The transverse diameter of the right hepatic lobe, caudate lobe-right lobe ratio, and splenic index were larger in patients with chronic schistosomiasis (p < 0.001). At multiple regression analysis, splenic siderotic nodules, splenic index, and caudate lobe-right lobe ratio were predictive of schistosomiasis. Observer agreement was substantial or almost perfect for almost all variables analyzed (kappa or r = 0.81-1.00). CONCLUSION: The presence of peripheral periportal fibrosis, heterogeneity of hepatic parenchyma, and splenic siderotic nodules, and the splenic index and caudate lobe right lobe ratio are useful features for differentiating alcoholic or virus induced cirrhosis from chronic schistosomiasis using MRI. PMID- 18287414 TI - Algorithm for immediate cytologic diagnosis of hepatic tumors. AB - OBJECTIVE: Immediate cytologic assessment of hepatic lesions can help determine the adequacy of specimens and may yield a preliminary diagnosis. The purpose of this study was to analyze the diagnostic accuracy of on-site cytologic assessment compared with definitive cytologic examination in the detection and correct subtyping of malignant hepatic lesions. SUBJECTS AND METHODS: The study population included 472 consecutively registered patients with hepatic nodules who underwent sonographically guided fine-needle aspiration and core biopsies. During on-site cytologic analysis, the pathologist made a preliminary diagnosis of malignancy or negative for malignancy for each nodule. When a malignant lesion was diagnosed, immediate subtyping was attempted. RESULTS: With immediate cytologic analysis, 280 (80.9%) of 346 malignant nodules were correctly identified without false-positive cases. With immediate subtyping, 113 (68.1%) of 166 cases of hepatocellular carcinoma, 28 (77.8%) of 36 cases of cholangiocarcinoma, and 113 (85.0%) of 133 cases of metastasis were detected. CONCLUSION: High diagnostic accuracy for malignancy can be achieved with on-site cytologic evaluation of hepatic tumors. Specific diagnoses of hepatocellular carcinoma, cholangiocarcinoma, and metastasis also can be made on-site in most cases. The absence of false-positive diagnosis of malignancy at on-site cytologic examination may make additional biopsy unnecessary. We propose an algorithm for the cytohistopathologic management of hepatic tumors. PMID- 18287415 TI - Clinical and economic impact of falsely decreased calcium values caused by gadoversetamide interference. AB - OBJECTIVE: Gadolinium is administered as a contrast agent in MRI procedures. Two gadolinium-based contrast agents, gadodiamide and gadoversetamide, interfere with colorimetric total serum calcium methods. The purpose of this prospective observational study was to examine the incidence of calcium interference after gadoversetamide procedures, associated clinical outcomes, and costs 20 months after implementation of quality assurance and physician education programs. MATERIALS AND METHODS: Records of patients who received gadoversetamide from June 24, 2006, to October 7, 2006, were reviewed to determine if a routine calcium test had been performed after the injection. Calcium values were repeated with an alternate method that is less susceptible to gadoversetamide interference. If the difference was > or = 2.0 mg/dL or if the initial test value was < or = 7.0 mg/dL, patient charts were reviewed for any related treatment. Costs associated with this algorithm were tracked. RESULTS: The initial calcium test was performed after gadoversetamide in 766 of 3,439 instances. The alternate test was performed in 633 of 766. One hundred twenty-five of 633 (20%) showed a difference in calcium values that was > or = 0.7 mg/dL, with 16 showing differences of > or = 1.6 mg/dL. Chart review for 56 instances revealed that calcium supplements were administered in 22 of 56 around the time of gadoversetamide injection. However, none appeared to be related to the spurious hypocalcemia. The total additional cost (reagent and technologist) for following this algorithm for just over 3 months was $6,807. CONCLUSION: Approximately 20% of patients receiving gadoversetamide exhibited spurious hypocalcemia. No patients were identified who received inappropriate calcium because of this interference. This may be attributable to the quality assurance and physician education programs. PMID- 18287416 TI - A step-by-step solution for embedding user-controlled cines into educational Web pages. AB - OBJECTIVE: The objective of this article is to introduce a simple method for embedding user-controlled cines into a Web page using a simple JavaScript. Step by-step instructions are included and the source code is made available. This technique allows the creation of portable Web pages that allow the user to scroll through cases as if seated at a PACS workstation. CONCLUSION: A simple JavaScript allows scrollable image stacks to be included on Web pages. With this technique, you can quickly and easily incorporate entire stacks of CT or MR images into online teaching files. This technique has the potential for use in case presentations, online didactics, teaching archives, and resident testing. PMID- 18287417 TI - Focal liver lesions: one more example of discordance between contrast-enhanced sonography and CT pattern of enhancement. PMID- 18287418 TI - Of hypes and hopes: N-acetylcysteine and cystatin C. PMID- 18287419 TI - National health care expenditure update: a new threat or an opportunity? PMID- 18287420 TI - "The roentgen manifestations of unrecognized skeletal trauma in infants"--a commentary. PMID- 18287421 TI - Back to the future. PMID- 18287422 TI - Optimism. PMID- 18287423 TI - MRI of metatarsal head subchondral fractures in patients with forefoot pain. AB - OBJECTIVE: The purpose of our study was to determine the MRI features of metatarsal head subchondral fractures in symptomatic adults. MATERIALS AND METHODS: A retrospective review of foot MRI procedures was performed to detect cases of metatarsal head subchondral fractures over a 6-year period. MR images of selected cases were analyzed by two reviewers for the presence of subchondral fracture, marrow edema-like pattern, metatarsal head flattening, and subchondral sclerosis. Patients with a history of foot surgery, infection, or inflammatory arthritis were excluded. Assessment for coexisting osseous and soft-tissue abnormalities was also performed. RESULTS: Subchondral fractures of the metatarsal heads were seen in 14 patients. All patients were women. The metatarsal head most commonly affected was the second (71%, 10/14) and the dorsal third of the metatarsal articular surface was involved in 79% (11/14). MRI findings of subchondral fracture of the metatarsal head with severe marrow edema like pattern were seen in 71% (10/14), suggesting early stage changes. Metatarsal head collapse with subchondral sclerosis and mild or absent marrow edema-like pattern were seen in 29% (4/14) indicating late-stage changes. Concurrent abnormalities included three patients (21%) with metatarsal shaft fractures and one patient (7%) with an interdigital neuroma. One subject was treated surgically. CONCLUSION: Subchondral fractures of the metatarsal heads can be detected on MR examinations of adults with forefoot pain. A subchondral fracture with associated marrow edema-like pattern is the most common presentation and likely reflects early stages of metatarsal head infraction. PMID- 18287424 TI - The snapping iliopsoas tendon: new mechanisms using dynamic sonography. AB - OBJECTIVE: The purpose of our study was to describe new mechanisms responsible for the snapping iliopsoas tendon using dynamic sonography. MATERIALS AND METHODS: We reviewed the video recordings obtained during dynamic sonography studies used to establish the diagnosis of 18 snapping iliopsoas tendons in 14 patients (nine females and five males; age range, 13-50 years) who presented clinically with either unilateral (n = 10) or bilateral (n = 4) snapping hips. During dynamic imaging, the transducer was positioned in a transverse oblique plane just above the hip joint parallel to the pubic bone. For all patients, the hip movement that generated the snapping consisted of bringing the hip from flexion-abduction-external rotation back to the neutral position. RESULTS: In 14 of 18 hips, the snapping was provoked by the sudden flipping of the iliopsoas tendon around the iliac muscle, allowing abrupt contact of the tendon against the pubic bone and producing an audible snap. Other causes of snapping iliopsoas tendon were bifid tendon heads flipping over one another (n = 3) and iliopsoas tendon impinging over an anterior paralabral cyst (n = 1). CONCLUSION: New mechanisms of snapping iliopsoas tendon have been described using dynamic sonography. Sudden iliopsoas tendon flipping over the iliac muscle was the most common cause of snapping hip. PMID- 18287425 TI - Imaging appearance of diffuse neurofibroma. AB - OBJECTIVE: The purposes of this study were to describe the imaging appearance of diffuse neurofibroma in 10 patients and to summarize demographic data on a large group of patients. MATERIALS AND METHODS: Retrospective review of the pathology and radiology teaching databases at two institutions yielded the cases of 339 patients with a pathologic diagnosis of diffuse neurofibroma. Diagnostic-quality images were available for 10 patients. Images from MRI (n = 8), CT (n = 5), and sonographic (n = 1) examinations were evaluated for lesion location, size, depth of involvement, growth pattern, and intrinsic signal intensity, attenuation, or echogenicity. Demographic information, associated lesions, and tumor location were recorded for all patients. RESULTS: Among 10 patients with images, eight of whom had neurofibromatosis, diffuse neurofibroma involved the skin and subcutaneous tissues (n = 9) and frequently extended to the fascia over muscle (n = 6). Plaquelike (n = 5) and infiltrative (n = 3) growth patterns were most common. One lesion had a mixed growth pattern. Prominent internal vascularity was common (n = 5). MRI signal intensity and CT attenuation were typically nonspecific. Enhancement was intense in all five patients with contrast-enhanced MR examinations. Including patients with and those without images, 349 diffuse neurofibromas were present in 339 patients. The mean patient age was 35.1 years. Lesions involved the extremities (n = 120), trunk (n = 122), head and neck (n = 98), and deep structures (n = 9). CONCLUSION: Diffuse neurofibroma frequently grows as a plaquelike or infiltrative lesion involving the skin and subcutaneous tissues. Prominent internal vascularity is common. There is a much wider soft tissue and age distribution and association with neurofibromatosis than previously reported. PMID- 18287426 TI - Sonography of the teres minor: a study of cadavers. AB - OBJECTIVE: The purpose of our study was to evaluate in cadavers the ability of high-resolution sonography to identify both the normal tendinous insertion and tears of the teres minor. MATERIALS AND METHODS: The teres minor insertion in five cadaveric shoulders was imaged, and methylene blue dye was injected into both the superior and inferior margins of the teres minor insertion by experienced musculoskeletal radiologists using a 10-5-MHz linear array transducer. Afterward, posterior shoulder dissection was performed. In another group of 11 cadaveric shoulders, an artificial tear was created at the teres minor insertion in six shoulders, and a sham procedure was performed in the remaining five shoulders arthroscopically. After arthroscopy, the teres minor insertion of each shoulder was imaged, and the accuracy of sonography for detecting a tear was evaluated. RESULTS: The dye was injected correctly into both the superior and inferior margins of the teres minor insertion in all five cadaveric shoulders. All six artificial tears were successfully detected on sonography. Four of the five specimens with the sham procedure were identified as having a normal teres minor insertion. One was misinterpreted as a tear. CONCLUSION: Sonography can reliably be used to identify the teres minor insertion and to detect tears of the teres minor muscle-tendon unit. PMID- 18287427 TI - Avulsion of the medial epicondyle after ulnar collateral ligament reconstruction: imaging of a rare throwing injury. AB - OBJECTIVE: The objective of this article is to report and describe the clinical and imaging features of an avulsion fracture of the medial epicondyle after ulnar collateral ligament (UCL) reconstruction. CONCLUSION: Avulsion fracture of the medial epicondyle is a rare complication of UCL reconstruction with distinct radiographic and MRI findings. PMID- 18287428 TI - Imaging and percutaneous treatment of secondarily infected hepatic infarctions. AB - OBJECTIVE: The objective of our study was to describe the imaging features and success rate of percutaneously treated infected hepatic infarctions. MATERIALS AND METHODS: Three hundred ninety-two patients had percutaneous liver abscess aspiration and drainage or aspiration and intraoperative debridement at our institution between 1990 and 2003. One hundred fifty-one of these patients underwent CT at least 2 days before the drainage procedure and immediately before the procedure. Retrospective review of the imaging and medical records identified 13 patients with microbiologically documented liver abscesses who had liver lesions consistent with hepatic infarction on the baseline CT. RESULTS: Twenty one hepatic infarctions in 13 patients were documented on baseline CT, 15 of which became secondarily infected. Ten of 15 patients with infected infarctions had undergone either hepatic transplantation or the Whipple procedure. Although the left lobe was slightly more commonly infarcted than the right lobe (54% vs 46%, respectively), right lobe infarctions were more commonly superinfected than left lobe infarctions (61% vs 39%); however, neither of these distinctions was statistically significant. Twelve of 13 patients underwent percutaneous drainage. The duration of catheter drainage was significantly longer in patients in whom catheter drainage was complicated by biliary communication than those without biliary communication (61 vs 19 days, respectively). Eleven of 12 patients (92%) responded to drainage such that they survived to discharge from the hospital. CONCLUSION: Patients with hepatic infarctions are at risk for secondary infection, particularly those patients having undergone surgery involving the porta hepatis. Percutaneous abscess drainage can be performed safely with excellent technical and clinical outcomes in this complex patient population. PMID- 18287429 TI - Chemoembolization of hepatocellular carcinoma: patient status at presentation and outcome over 15 years at a single center. AB - OBJECTIVE: We report the outcome of the care of 209 patients with hepatocellular carcinoma with a focus on relevant scoring systems for predicting overall survival and time to progression and on changes in presentation status and outcome from 1991 to 2006. MATERIALS AND METHODS: Hepatic arterial chemoembolization was performed on 209 patients in 375 sessions. Disease status was evaluated with the Child-Pugh, Okuda, Cancer of the Liver Italian Program, and American Joint Committee on Cancer (AJCC) systems. Changes in status at presentation from 1991 to 2006 and change in overall survival period and time to progression were analyzed. RESULTS: Median and mean overall survival periods for the entire group were 376 and 574 +/- 61 days. Median and mean times to progression were 267 and 409 +/- 54 days. Forty-nine patients underwent liver transplantation a median of 143 days after chemoembolization. The median and mean overall survival times among patients not undergoing transplantations were 466 and 574 +/- 61 days. Okuda score (p < 0.0001) and AJCC stage (p = 0.014) were the best predictors of overall survival and time to progression, respectively. Patients with disease with an Okuda I score and in AJCC stage I or II had median and mean overall survival periods of 667 and 992 +/- 176 days and times to progression of 378 and 589 +/- 110 days. Clinical status at presentation, overall survival period (p = 0.64), and time to progression (p = 0.44) were unchanged from 1991 to 2006. The 30-day mortality was 3.2%. CONCLUSION: Patients treated with hepatic arterial chemoembolization for HCC in Okuda score I and AJCC stage I or II have more durable survival than previously reported in a U.S. population. PMID- 18287430 TI - Imaging-guided percutaneous needle aspiration or catheter drainage of neonatal liver abscesses: 14-year experience. AB - OBJECTIVE: The purpose of our study was to review the clinical aspects and long term outcomes of imaging-guided percutaneous aspiration or drainage of liver abscesses in neonates. MATERIALS AND METHODS: We retrospectively reviewed the clinical and imaging records of eight neonates with liver abscesses referred for imaging-guided percutaneous aspiration or drainage, including one autopsy-proven case in whom the percutaneous aspiration or drainage was not performed. Clinical and imaging features, complications, and long-term follow-up results were assessed. RESULTS: Eight neonates with liver abscesses were referred for imaging guided percutaneous aspiration or drainage (five males, three females; age range, 7-100 days; weight, 610-3,400 g). Six were born prematurely (24-29 weeks of gestation). Six had a history of umbilical catheterization. All were clinically septic. All neonates received long-term i.v. antibiotics. Five neonates had solitary multiloculated abscesses (right lobe [n = 3], straddling both lobes [n = 2]), and three had solitary uniloculated abscesses (right lobe [n = 1] and left lobe [n = 2]). Imaging-guided drainage catheter insertion (n = 4), aspiration (n = 2), and aspiration followed by drainage catheter insertion (n = 1) were performed in seven neonates within 1 day after referral. Coagulase-negative Staphylococcus (4/8) was the most common organism isolated from blood and pus. There were no procedure-related complications. Catheter repositioning was required in one. Serial sonography (mean, 12.5 months) and clinical follow-up (mean, 20.7 months) showed complete clinical remission in seven cases. Three healed with calcification in the previous abscess site. Long-standing left portal vein thrombosis was seen in two cases. CONCLUSION: Neonatal liver abscess is associated with good long-term outcome and minimal complications when imaging guided percutaneous aspiration or drainage is performed in conjunction with long term antibiotic coverage. PMID- 18287431 TI - Which patients benefit from a 3D reconstructed coronal view of the uterus added to standard routine 2D pelvic sonography? AB - OBJECTIVE: The objective of our study was to evaluate whether a 3D reconstructed coronal view of the uterus provides added benefit to standard gynecologic sonography. MATERIALS AND METHODS: Sixty-six consecutive patients underwent standard 2D pelvic sonography followed by 3D sonography. The physician determined whether the reconstructed coronal view of the uterus was helpful to make a diagnosis not possible with the 2D scan, helpful to be more confident of a diagnosis suspected on the basis of the 2D scan, or not helpful. Comparison of the demographic information, sonographic findings, and endometrial thickness was made between the patient groups. RESULTS: The 3D coronal views of the uterus added value to the 2D scan in 16 (24%) of the 66 patients. In five of these 16 patients, the coronal view added information about findings not seen using 2D imaging. In the other 11 patients, the diagnostic findings were more confidently seen using the coronal view. The coronal view added no information in 50 patients. The coronal view was helpful in four (12.5%) of 32 patients with an endometrium < 5 mm, one of six patients whose endometrium was incompletely seen with 2D sonography, and 11 (39%) of 28 patients whose endometrium measured > or = 5 mm. The coronal view did not provide benefit in patients who had normal findings on 2D scanning. In three patients referred because of infertility, uterine shape anomalies were diagnosed using the coronal view. CONCLUSION: The 3D coronal view of the uterus is a valuable adjunct to a 2D pelvic scan, particularly in patients presenting with infertility or suspected endometrial lesions. In addition, the coronal view is helpful in patients with an endometrium > or = 5 mm. PMID- 18287432 TI - Papillary lesions of the breast diagnosed at percutaneous sonographically guided biopsy: comparison of sonographic features and biopsy methods. AB - OBJECTIVE: The objective of our study was to retrospectively assess the potential role of sonographic features and biopsy in the management of patients with papillary lesions diagnosed at percutaneous sonographically guided biopsy. MATERIALS AND METHODS: Surgical or sonographic follow-up (> or = 2 years) was available in 123 women (age range, 21-75 years; mean age, 47 years) with 124 papillary lesions diagnosed at sonographically guided core needle or vacuum assisted biopsy during a 7-year period. Surgical excision results or follow-up sonograms with no change at the 2-year follow-up served as the reference standard. We reviewed the sonographic features, biopsy results, and surgical or sonographic follow-up. Statistical analysis was performed using the Fisher's exact test for the difference of sonographic features. RESULTS: Cancer incidence per BI-RADS category was as follows: category 3, zero (0%) of 21; category 4a, 14 (17%) of 85; category 4b, four (36%) of 11; category 4c, one (20%) of five; and category 5, one (50%) of two. On sonography, two features distinguishing benign from malignant papillary lesions were echo pattern and margins of masses. Core needle biopsy gave a sensitivity of 28% and specificity of 100%, whereas both sensitivity and specificity were 100% with vacuum-assisted biopsy. Of 117 lesions with benign biopsy results, upgrade to malignancy or high-risk lesion was found in 17% of benign papillomas, 24% of atypical papillomas, and 0% of multiple papillomas. CONCLUSION: Vacuum-assisted biopsy was more accurate than core needle biopsy in diagnosing papillary lesions, and both sonographic features and core needle biopsy were not sufficiently accurate. Therefore, surgical excision should be performed for the accurate diagnosis of papillary lesions. PMID- 18287433 TI - Lobular neoplasia at percutaneous breast biopsy: variables associated with carcinoma at surgical excision. AB - OBJECTIVE: The purpose of our study was to better define the rate and variables associated with cancer underestimation when lobular neoplasia is found at minimally invasive breast biopsy. MATERIALS AND METHODS: The records of 32,420 patients who underwent imaging-guided needle biopsy of the breast for mammographic or sonographic abnormalities from 1988 to 2000 were retrospectively reviewed. The 278 cases in which lobular neoplasia was the highest-risk lesion at biopsy were included. Of the 278 cases, 164 proceeded to surgical excision, allowing calculation of rates of underestimation from minimally invasive biopsy. RESULTS: Of the 32,420 minimally invasive breast biopsies, lobular neoplasia was found in 278 (0.9%). One hundred sixty-four of the 278 (59%) continued to surgical excision, where cancer was pathologically confirmed in 38 (23%). No difference was seen in the underestimation rates for lesions diagnosed as lobular carcinoma in situ (25%, 17 of 67 lesions) versus atypical lobular hyperplasia (22%, 21 of 97 lesions). Statistically significant underestimation of carcinoma was found with biopsy of masses (with or without associated microcalcifications) rather than calcifications only, a higher BI-RADS category (p < 0.0001), use of a core biopsy device rather than a vacuum device (p < 0.01), and obtaining fewer specimens (p < 0.0001). CONCLUSION: Significant sampling error occurs regardless of the type of core biopsy device, number of specimens obtained, histologic radiographic concordance, mammographic appearance, and complete excision of the lesion as determined by imaging. For this reason, all patients with lobular neoplasia at core or vacuum-assisted biopsy should undergo surgical excision until further differentiating criteria can be determined. PMID- 18287434 TI - MDCT of acute mild (nonnecrotizing) pancreatitis: abdominal complications and fate of fluid collections. AB - OBJECTIVE: The objective of our study was to describe the occurrence of local complications and the fate of fluid collections in milder forms of acute nonnecrotizing pancreatitis. MATERIALS AND METHODS: Initial MDCT studies of 169 consecutive patients with mild acute pancreatitis and 203 follow-up CT examinations were reviewed. The fate of peripancreatic fluid collections was investigated, and the incidence and type of local complications were recorded and correlated to the CT grading system (A-E). RESULTS: Complications developed in nine of 169 patients, for an incidence of 5.3%. All morbidity occurred in the subgroup of 73 patients with initial fluid collections, for an incidence of 12.3%. Follow-up CT examinations available in 51 of these 73 patients documented rapid fluid resolution in 35 cases (68.6%) and persistence of fluid more than 2 weeks from onset in seven asymptomatic patients (13.7%). Acute, life-threatening complications (hemorrhage, infection, perforation) occurred in five patients, for an incidence of 6.8% among the 73 patients with initial fluid collections, or 3.0% in the entire group of 169 patients. Five patients developed acute pseudocysts. Long-term follow-up studies discovered two patients with chronic pancreatitis and one with groove pancreatitis. CONCLUSION: A small number of acute, life-threatening abdominal complications and chronic complications are expected to occur in patients with milder forms of acute nonnecrotizing pancreatitis presenting with fluid collections. In these patients, clinical monitoring and repeated imaging studies are recommended to document the resolution of fluid or the development of complications. PMID- 18287435 TI - Reversed diastolic flow in the renal transplant: perioperative implications versus transplants older than 1 month. AB - OBJECTIVE: The purpose of our study was to evaluate the causes, waveform morphology, and clinical outcomes of high-resistance reversed diastolic flow in transplanted kidneys. MATERIALS AND METHODS: To identify patients with reversed diastolic flow, we performed a review of 5,089 renal transplant Doppler sonograms obtained over a 10-year period. Waveform morphology was correlated with surgical histologic findings and clinical outcomes. RESULTS: Fifty-nine patients (33 male, 26 female; age range, 14-69 years) with reversed diastolic flow fell into three chronologic groups: acute group (six patients), transplant < 24 hours; perioperative group (34 patients), transplant < or = 30 days; and long-term group (19 patients), transplant > 30 days. Acute reversed diastolic flow was associated with higher likelihood of graft survival (p = 0.001, Fisher's exact test) compared with reversed diastolic flow discovered in the perioperative or long term group. In the acute group, hematoma, acute tubular necrosis, renal vein thrombosis, and vascular kink produced reversed diastolic flow. The causes of reversed diastolic flow for the perioperative group were acute tubular necrosis, rejection, and renal vein thrombosis; for the long-term group, reasons for diastolic reversal were rejection, glomerulosclerosis, low cardiac output, and diabetic nephrosclerosis. The causes of reversed diastolic flow were not differentiated by waveform morphology. CONCLUSION: The causes of reversed diastolic flow cannot be distinguished by waveform morphology. Patients with reversed diastolic flow < 24 hours after transplantation warrant emergent exploration because correction of treatable causes may lead to recovered function. Long-standing renal transplants with reversed diastolic flow are not likely salvageable. PMID- 18287436 TI - Mucinous appendiceal neoplasms: preoperative MR staging and classification compared with surgical and histopathologic findings. AB - OBJECTIVE: The objective of our study was to determine the accuracy of MRI in the preoperative staging and classification of mucinous appendiceal neoplasms and to describe the MRI features that are useful for selecting patients for surgical resection. MATERIALS AND METHODS: Twenty-two patients underwent preoperative MRI including T1-weighted, T2-weighted, immediate gadolinium-enhanced, and delayed gadolinium-enhanced imaging. Two observers reviewed the images for peritoneal tumor at 13 sites, tumor size and distribution, and degree of tumor enhancement. Peritoneal tumor sites were recorded at surgery. Cytoreduction was categorized as complete or suboptimal. Surgical specimens were classified as disseminated peritoneal adenomucinosis tumors, intermediate-grade tumors, or peritoneal mucinous carcinomatosis tumors. RESULTS: Surgery confirmed 232 tumor sites. Delayed gadolinium-enhanced MRI was the most accurate of the MR techniques, with a sensitivity, specificity, and accuracy of 89%, 87%, and 89%, respectively, for observer 1 and 82%, 87%, and 83% for observer 2 (p < 0.001). Surgical cytoreduction was complete in 14 patients and suboptimal in eight. MRI findings predicting suboptimal cytoreduction included a large (> 5 cm) mesenteric mass, which was present in 75% of the patients in the suboptimal cytoreduction group and 0% of those in the complete cytoreduction group; diffuse mesenteric tumor (88% and 0%, respectively); tumor encasement of mesenteric vessels (88% and 0%); or diffuse small-bowel serosal tumor (75% and 0%). Histopathology results showed six disseminated peritoneal adenomucinosis tumors, four intermediate tumors, and 11 peritoneal mucinous carcinomatosis tumors. The specimens for the remaining patient were not available for histopathologic analysis. Qualitatively, the 11 peritoneal mucinous carcinomatosis tumors showed greater enhancement than the liver, whereas six disseminated peritoneal adenomucinosis and the four intermediate tumors showed less enhancement than the liver. Quantitatively, the mean tumor-to-liver contrast for disseminated peritoneal adenomucinosis and intermediate tumors was 0.67 compared with 1.53 for peritoneal mucinous carcinomatosis tumors (p < 0.0001). CONCLUSION: Of the MR techniques evaluated, delayed gadolinium-enhanced MRI was the most accurate for the staging and classification of mucinous appendiceal neoplasms and provided prognostic information useful for patient selection. PMID- 18287437 TI - Anaphylaxis to iodinated contrast material: nonallergic hypersensitivity or IgE mediated allergy? AB - OBJECTIVE: Contrast material is generally well tolerated although approximately 1% of patients who receive low-osmolar nonionic contrast material will develop anaphylaxis symptoms. Because most anaphylactic reactions are mild and nonallergic, clinically mimicking immunoglobulin E (IgE)-mediated allergy, diagnostic skin testing has been discussed controversially in the past and prophylactic pretreatment drug regimens are recommended instead. In the past 6 years, all patients with contrast material-induced anaphylaxis have been subjected to allergologic diagnostic procedures to clearly differentiate allergic and nonallergic anaphylaxis. Thus the purpose of our study was to identify and differentiate IgE-mediated allergy and nonallergic contrast material-induced hypersensitivity. Furthermore, the objective of our diagnostic procedures was not only to identify the culprit contrast material but also to find alternative contrast material for future radiologic interventions. SUBJECTS AND METHODS: We evaluated 96 patients with anaphylaxis symptoms after contrast material application using standardized intradermal skin testing. In patients with positive skin tests, the IgE-mediated allergy was further evaluated with in vitro and challenge tests. RESULTS: In four patients (suffering from anaphylaxis grades 2 and 3) out of the 96 (4.2%), skin tests and basophil activation tests strongly suggested IgE-mediated allergy to the contrast materials iopromide (two patients), iomeprol, and iopentol. In two patients with allergies to iopromide and iomeprol, alternative nonionic monomer contrast materials were tolerated, as identified in controlled challenge tests with iopamidol and iopromide, respectively. CONCLUSION: The evaluation of patients with contrast material induced anaphylaxis (at least those with anaphylaxis > or = grade 2) should always include appropriate skin tests ensuring that patients with an IgE-mediated allergy are not missed. Moreover, allergologic testing may identify a contrast material of the group of nonionic monomers that will be tolerated in future radiologic interventions. PMID- 18287438 TI - Cross-sectional imaging of the anal sphincter in fecal incontinence. AB - OBJECTIVE: Fecal incontinence is a disabling disorder. Cross-sectional imaging techniques can be used to confirm the diagnosis and to clarify the anatomy and function of the anorectal region. CONCLUSION: Cross-sectional imaging has increased the understanding of the sphincter complex, resulting in a more adequate evaluation of fecal incontinence. PMID- 18287439 TI - Benefits of contrast-enhanced sonography for the detection of liver lesions: comparison with histologic findings. AB - OBJECTIVE: The objective of our study was to compare the usefulness of contrast enhanced sonography with baseline sonography in detecting malignant liver lesions. SUBJECTS AND METHODS: This prospective study included 116 patients. All patients underwent a preoperative conventional sonography examination followed by sonography after injection of contrast agent combined with the use of perfusion software (vascular recognition imaging or pulse subtraction imaging). Histopathologic analysis was the reference standard used to compare the diagnostic value of baseline sonography versus contrast-enhanced sonography. RESULTS: Eighty-two patients underwent hepatic surgery, 31 did not because of disseminated lesions, and the remaining three patients did not meet inclusion criteria. Three hundred six surgically proven lesions were taken into account for comparison of the two techniques: 147 were detected on baseline sonography and 177 on contrast-enhanced sonography. Histopathologic analysis revealed 233 malignant and 73 benign lesions. Sensitivity and specificity were improved on contrast-enhanced sonography compared with baseline sonography for the detection of malignant lesions: 68.7% versus 58.8% and 67% versus 50.7%, respectively. Contrast-enhanced sonography detected 23 additional malignant lesions that had been seen as lacuna at the portal venous phase and characterized as 19 benign nodules, thus improving the performance of sonography in 13.7% of the cases. CONCLUSION: Contrast injection improved the sensitivity and specificity of baseline sonography and should be performed in routine practice if hepatic surgery is being considered for the management of liver lesions. PMID- 18287440 TI - Real-time temporal maximum-intensity-projection imaging of hepatic lesions with contrast-enhanced sonography. AB - OBJECTIVE: We sought to perform a preliminary evaluation of temporal maximum intensity projection (MIP) of focal hepatic masses in selected patients. The technique processes real-time contrast-enhanced sonography images by integrating the path of moving bubbles to depict vascular morphology. Following a high intensity ultrasound pulse that disrupts bubbles within the scan plane, MIP images the trajectories of fresh bubbles replenishing the plane and revealing their course. CONCLUSION: Vascular morphology is depicted at a level or detail not seen before with sonography. High-frame-rate sequences of less than one second uniquely show arterial structure in liver lesions. PMID- 18287441 TI - Automated carbon dioxide insufflation for CT colonography: effectiveness of colonic distention in cancer patients with severe luminal narrowing. AB - OBJECTIVE: The objective of our study was to determine the effectiveness of automated CO2 insufflation in colonic distention for CT colonography (CTC) in patients with severe luminal narrowing by colorectal cancer and preliminarily evaluate its safety performed shortly after colonoscopic polypectomy or biopsy. MATERIALS AND METHODS: Seventy-four patients were examined with colonoscopy and subsequent CTC (time interval, 0-8 days) using automated CO2 insufflation. Thirty six patients whose colonoscopy was incomplete due to severe luminal narrowing by cancer that prevented colonoscope passage constituted the stenotic group. The remaining 38 patients constituted the nonstenotic group. Colonic distention was graded by two experienced readers from 1 (worst) to 4 (best) and compared between the two groups. Clinical data and CT images were analyzed for the occurrence of colonic perforation. RESULTS: Distention was not significantly different between the stenotic and nonstenotic groups in any colonic segments in both supine and prone positions. The mean distention grade +/- SD of the colonic segments proximal to the luminal narrowing in the stenotic group (n = 143 segments) was 3.7 +/- 0.7 and 3.8 +/- 0.7 for the supine and prone positions, respectively. Colonic perforation was not noted in any of the 74 patients, including 65 patients who underwent CTC within 24 hours after colonoscopy (62 snare polypectomies, two polypectomies using biopsy forceps, 63 routine mucosal biopsies). CONCLUSION: Automated pressure-controlled CO2 insufflation is as efficient in colonic distention for CTC in colorectal cancer patients with severe luminal narrowing as it is in patients without severe luminal narrowing. PMID- 18287442 TI - Dropped appendicolith: CT findings and implications for management. AB - OBJECTIVE: The aim of this study was to discern the CT features of appendicoliths retained after appendectomy and evaluate the management options. CONCLUSION: Retained, or dropped, appendicolith most often presents as an area of high attenuation less than 1 cm in diameter with an associated abscess close to the cecum or Morison's pouch. CT-guided drainage can be used to manage abscesses associated with dropped appendicolith in selected cases. PMID- 18287443 TI - CT of gastrointestinal complications associated with hematopoietic stem cell transplantation. AB - OBJECTIVE: This article illustrates the characteristic CT findings of bowel damage related to drug toxicity, conditioning regimen, infections, and graft versus-host disease (GVHD) in patients after hematopoietic stem cell transplantation. CONCLUSION: Although some overlap exists in the CT appearances of the different causes of bowel wall inflammation, the findings are frequently unique enough to suggest a specific diagnosis. Awareness of the specific time of occurrence, intensity of clinical symptoms, preferred localization, and extent of accompanying extraintestinal findings help to distinguish the different pathologic entities. PMID- 18287444 TI - Characterization of human renal stones with MDCT: advantage of dual energy and limitations due to respiratory motion. AB - OBJECTIVE: Our aim was to determine, using CT attenuation values, the chemical composition of 241 human renal stones placed in a jelly phantom and to analyze the influence of respiratory motion on the classification. MATERIALS AND METHODS: The stones were placed in a jelly simulating the X-ray attenuation of the kidneys. A dynamic platform was used to apply to the phantom free-breathing motion (sinusoidal motion in z-axis) and motion due to lack of maintenance of a breath-hold (5 mm x s(-1) in z-axis). Determination of the chemical composition was performed with mean CT attenuation values obtained at 80 and 120 kV and with dual-energy CT attenuation values. RESULTS: Two hundred forty-one human urinary stones were classified into six groups: uric acid, cystine, struvite, weddellite (calcium oxalate dihydrate), whewellite (calcium oxalate monohydrate), and brushite. With no motion, the use of dual energy enabled differentiation of all of the types of stones with statistically significant differences. Uric acid (-20 +/- 22 H), cystine (106 +/- 19 H), struvite (271 +/- 16 H), weddellite (323 +/- 5 H), brushite (415 +/- 30 H), and whewellite (510 +/- 17 H) were identified as distinct groups. Motion-induced mean CT attenuation values were significantly different from those obtained with no motion. With motion, dual-energy CT attenuation values did not allow differentiation of all stone types. CONCLUSION: Dual-energy CT attenuation values can be used to predict the chemical composition of stones in vitro. However, when slight motion is applied to renal stones during image acquisition, the values become significantly different from those obtained with no motion. Consequently, confusion arises in differentiating stone types. A perfect breath-hold has to be performed for in vivo use of attenuation value to discern stone type. PMID- 18287445 TI - 16-MDCT cystoscopy in the evaluation of neoplasms of the urinary bladder. AB - OBJECTIVE: The purpose of this study was to evaluate the utility of 16-MDCT cystoscopy in the detection of urinary bladder neoplasms in a high-risk population. SUBJECTS AND METHODS: Fifty patients who presented with hematuria and a recent diagnosis or a history of bladder carcinoma underwent CT cystoscopy. All patients were examined in the supine and prone positions after bladder distention with room air. A detector configuration of 16 x 0.75 mm and a pitch of 1.2 was used. Virtual images were obtained with volume-rendered algorithms. Transverse tomographic slices, multiplanar reformatted images, and virtual images were prospectively evaluated separately and in combination. Conventional cystoscopy was considered the standard of reference for assessing the efficacy of MDCT cystoscopy in the detection of urinary bladder tumors. RESULTS: Fifty-five (96%) of 57 urinary bladder lesions recognized at conventional cystoscopy were detected with MDCT cystoscopy. The size of the lesions ranged from 0.3 to 9.7 cm in diameter, including 18 lesions with a diameter of 0.5 cm or less. Transverse, multiplanar reformatted, and virtual images proved complementary for lesion detection. CONCLUSION: MDCT cystoscopy is an accurate technique for the detection of urinary bladder neoplasms in patients at high risk, yielding satisfactory results in the identification of lesions smaller than 0.5 cm. PMID- 18287446 TI - Nephrogenic systemic fibrosis: a report of 29 cases. AB - OBJECTIVE: The purpose of this study was to determine the incidence of nephrogenic systemic fibrosis and its relation to renal failure and the administration of gadolinium-based contrast material at an academic medical center. MATERIALS AND METHODS: A dermatopathology database was searched to identify patients in whom nephrogenic systemic fibrosis was diagnosed. The medical records of these patients were reviewed. Renal function concurrent with any administration of gadolinium-based contrast material was assessed, as was patient outcome. A database of patients undergoing long-term dialysis was reviewed separately to determine how many had received gadolinium and the frequency of nephrogenic systemic fibrosis among these patients. RESULTS: Twenty nine patients were found to have had nephrogenic systemic fibrosis between November 15, 1999, and December 31, 2006. It was known that gadolinium-based contrast material had been administered to 25 of these patients before diagnosis. All 29 patients had compromised renal function (27 had chronic renal failure, and two had acute renal failure). Determination of the temporal relation between gadolinium-based contrast administration and symptom onset often was difficult. Only eight patients had severe morbidity. Nephrogenic systemic fibrosis developed in 12 (2.9%) of 414 patients undergoing long-term dialysis who received gadolinium-based contrast material. CONCLUSION: We confirm the strong association between nephrogenic systemic fibrosis and gadolinium-based contrast administration. Although the use of high doses of gadolinium and the occurrence of chronic renal failure have been implicated in other reports, several of our patients received standard doses of gadolinium, and two had transient acute renal failure before diagnosis. Most patients had mild or moderate symptoms. Nephrogenic systemic fibrosis developed in 2.9% of patients undergoing long-term dialysis who received gadolinium-based contrast material but in none of the long term dialysis patients who did not receive gadolinium-based contrast material. PMID- 18287447 TI - 64-MDCT angiography of the coronary arteries: nationwide survey of patient preparation practice. AB - OBJECTIVE: The purpose of this study was to evaluate the current practice of patient preparation for 64-MDCT angiography (CTA) of the coronary arteries. MATERIALS AND METHODS: Sites in the United States that perform 64-MDCT coronary angiography were surveyed by mail in 2006. Information requested included physician specialty; experience level; details about patient preparation, including the use, dose, route, and timing of premedication; and acceptable heart rate and rhythm. A total of 142 surveys were analyzed, with comparison of parameters across specialties (radiology, cardiology, or shared) and experience levels. RESULTS: All facets of the study (premedication, data acquisition, cardiac interpretation) are performed exclusively by radiologists in 49% of sites and by cardiologists in 14%. All sites administer beta-blockers. Target heart rate was reported as < or = 65 beats per minute (bpm) by 89% of responders. Despite most centers aiming for a heart rate of < or = 65 bpm, the maximum allowable heart rate is > 65 bpm in 80% of centers. Patients with arrhythmia are scanned in at least 25% of sites. Most sites (84%) administer nitroglycerin. Significant differences between specialties were noted for experience levels, timing and route of beta-blocker administration, and for target heart rate. The likelihood of scanning in the setting of arrhythmia and beta-blocker timing correlated with experience levels. CONCLUSION: These 64-MDCT coronary artery data from 2006 reveal consensus for a range of patient preparation parameters. Use of beta-blockers and nitroglycerin is routine, and the target heart rate is usually < or = 65 bpm. However, differences were noted for beta-blocker protocols and acceptable heart rate and rhythm, and some differences in practice are associated with experience level and specialty. PMID- 18287448 TI - Identification and quantification of coronary atherosclerotic plaques: a comparison of 64-MDCT and intravascular ultrasound. AB - OBJECTIVE: The purpose of this study was to compare the accuracy of 64-MDCT with that of intravascular ultrasound (IVUS) for the identification and quantitative analysis of coronary atherosclerotic plaques. MATERIALS AND METHODS: Twenty-six patients (17 men, nine women; mean age, 56 years) with suspected coronary atherosclerotic disease were studied using contrast-enhanced 64-MDCT and IVUS. The coronary arteries were divided into 10-mm segments and vascular cross sectional area (CSA), luminal CSA, and plaque burden were measured in each segment. Plaque analysis software was used to automatically detect both plaques and vessel walls on CT images. Two investigators who were blinded to IVUS results independently determined the presence, classification, and quantitative measurement of atherosclerotic plaques on the CT images, which were then compared with the IVUS images. RESULTS: Of 40 coronary arteries, 247 of 263 segments were imaged and analyzed by both contrast-enhanced 64-MDCT and IVUS. Sixteen segments were ruled out because of poor CT image quality. Compared with IVUS, 64-MDCT enabled correct detection in 86 of 89 (96.6%) segments containing noncalcified plaques, 25 of 27 (92.6%) segments containing calcified plaques, and 118 of 131 (90.1%) segments without atherosclerotic plaques. Sensitivity, specificity, positive predictive value, and negative predictive value for the detection of plaques by 64-MDCT were 97.4%, 90.1%, 89.7%, and 97.5%, respectively. Plaque analysis software using predetermined Hounsfield unit ranges for different components of plaque was able to distinguish between fibrous, fibrous-soft, and calcified plaques to a significant degree, but was less able to distinguish between soft and fibrous, and between soft and fibrous-soft plaque. Cohen's kappa coefficient for the sole detection of atherosclerotic segments by observers was 0.91. The correlation coefficients to determine vascular CSA, luminal CSA, and plaque burden were r = 0.85, 0.82, and 0.77, respectively (p < 0.01). CONCLUSION: Compared with IVUS, contrast-enhanced 64-MDCT has a good ability to identify and quantify coronary atherosclerotic plaques. However, the reliable differentiation of the composition of noncalcified plaques is still limited. PMID- 18287449 TI - Screening for lung cancer. AB - OBJECTIVE: The purpose of our review is to discuss the current state of lung cancer screening using CT in the context of defined criteria for effective screening. CONCLUSION: Although there are hopeful developments in lung cancer screening, a number of unresolved issues must be answered before adopting screening on a large scale. Currently no data exist to suggest that lung cancer screening with CT will result in a decrease in lung cancer mortality. PMID- 18287450 TI - Quantitative assessment of air trapping in chronic obstructive pulmonary disease using inspiratory and expiratory volumetric MDCT. AB - OBJECTIVE: The purpose of our study was to determine the attenuation threshold value for the detection and quantification of air trapping using paired inspiratory and expiratory volumetric MDCT scans and to assess whether the densitometric parameter can be used for the quantification of airway dysfunction in chronic obstructive pulmonary disease (COPD) regardless of the degree of emphysema. MATERIALS AND METHODS: This study included 36 patients with COPD who underwent 64-MDCT. The entire lung volume with attenuation between -500 and 1,024 H was segmented as whole lung. The lung volume with attenuation between 500 and -950 H was segmented as limited lung, while the lung volume of less than 950 H was segmented as emphysema and eliminated. The relative volumes for limited lung (relative volume(n-950)) with attenuation values below thresholds (n) ranging from -850 to -950 H, and relative volume for whole lung (relative volume( 15%), relative volume change(860 950) significantly correlated with the results of PFTs, whereas no significant correlations were seen between relative volume change(<-860) and PFTs. In the minimal or mild emphysema group (inspiratory relative volume(<-950) < 15%), all densitometric parameters correlated with PFTs. CONCLUSION: The densitometric parameter of relative volume change calculated on paired inspiratory and expiratory MDCT using the threshold of -860 H in limited lung correlated closely with airway dysfunction in COPD regardless of the degree of emphysema. PMID- 18287451 TI - MR angiography of lower extremities at 3 T: presurgical planning of fibular free flap transfer for facial reconstruction. AB - OBJECTIVE: The purpose of our study was to evaluate the use of preoperative MR angiography of the lower extremities at 3 T in candidates for fibular free flap harvesting, identifying atherosclerotic occlusive disease and congenital anomalies in this population. Our intention was to document the influence of the imaging findings on the surgical approach used. MATERIALS AND METHODS: Twenty nine consecutive adult patients with facial abnormalities necessitating mandibular resection with subsequent osteocutaneous mandibular reconstruction who underwent preoperative MR angiography at 3 T were retrospectively reviewed. Images were evaluated by two observers with regard to image quality and visualization of arterial segments; severity of stenosis; and presence of noise, artifact, or venous contamination. The popliteal artery branching pattern present was also classified. The facial and reconstructive surgeon involved indicated whether the MR angiographic appearances influenced the decision regarding the side or location from which the flap was harvested or the flap design. RESULTS: Arterial segments were visualized with good or excellent image quality in 722 of 725 segments for observer 1 and 721 segments for observer 2. The kappa coefficient indicated good interobserver agreement (kappa = 0.78) with regard to quality of arterial segment depiction and scoring of stenoocclusive disease (kappa = 0.64). No segments had venous contamination, noise, or artifact of a degree sufficient to compromise diagnostic interpretation. Imaging influenced the surgical approach in 16 (55.2%) of 29 patients. CONCLUSION: Trifurcation vessel imaging should be a prerequisite to fibular free flap harvesting. High-spatial resolution MR angiography at 3 T represents a desirable alternative to other invasive or cross-sectional imaging techniques in this regard. PMID- 18287452 TI - Contrast enhancement in cardiovascular MDCT: effect of body weight, height, body surface area, body mass index, and obesity. AB - OBJECTIVE: The purpose of our study was to evaluate the effect of body weight, height, body surface area (BSA), body mass index (BMI), and obesity on aortic contrast enhancement in cardiac MDCT. MATERIALS AND METHODS: Seventy-three consecutive patients underwent cardiac CT angiography on a 64-MDCT scanner. Seventy-five mL of contrast medium (350 mg I/mL) was injected at 4.5 mL/s, followed by a 40-mL saline flush at 4.5 mL/s. The scanning delay of CT was determined with a bolus tracking technique. Aortic attenuation was measured over the aortic-root lumen. BMI and BSA were calculated from the patient's body weight and height. The patients were divided into low-(BMI < 30) and high-(> or = 30) BMI groups. Associations of aortic attenuation with body weight, height, BMI, and BSA were evaluated with regression analysis and the Student's t test. RESULTS: Strong inverse correlations were seen between aortic attenuation and body weight (r = -0.73), height (r = -0.47), BMI (r = -0.63), and BSA (r = -0.74) (p < 0.001 for all). The regression formula of aortic attenuation versus body weight suggests that 1.0 mL/kg of contrast medium would yield a mean aortic attenuation of 355 H. The mean aortic attenuation was significantly higher in the low-BMI (352.6 +/- 59.1 H) than in the high-BMI (286.2 +/- 55.5 H) group. The regression formula for aortic attenuation on body weight was aortic attenuation = 586-3.1 body weight (p < 0.001) for the low-BMI group and aortic attenuation = 485-1.9 body weight (p < 0.001) for the high-BMI group, suggesting that the amount of contrast medium required with increased body weight is less in the high-BMI group. This group difference was less pronounced for the regression of aortic attenuation on BSA. CONCLUSION: To achieve a consistent contrast enhancement in cardiac CT angiography (CTA), contrast-medium dose should be adjusted with the body weight or the BSA (which accounts for both the body weight and height factors) to provide adjustment of iodine dose over a wide range of body sizes. PMID- 18287453 TI - Thin-slice MDCT of the neck: impact on cancer staging. AB - OBJECTIVE: The objective of this study was to compare thin-slice multiplanar evaluation and conventional 3-mm axial evaluation of head and neck MDCT in tumor staging. MATERIALS AND METHODS: Ninety-six patients with histologically proven squamous cell carcinoma were evaluated independently, once using 3-mm axial images and once using 1-mm interactive multiplanar reformation (MPR) images. Tumor stage was assessed with both methods; histology served as the reference. Thirty-seven patients with hypopharyngeal and laryngeal tumors had en bloc resection, allowing direct comparison of tumor infiltration into designated anatomic structures. Two examiners independently assessed the data sets. Interobserver agreement was tested with a modified kappa test. The Wilcoxon signed rank test with continuity correction was applied to test the null hypothesis, which postulates the equality of both methods. The chi-square test was applied to compare the number of correctly classified tumors for the two methods and readers. RESULTS: Interobserver agreement was high (kappa = 0.88 0.91). Both methods allowed accurate tumor staging, and no significant differences between the two methods were found (reader A, p = 0.61; reader B, p = 1). With MPR assessment, more anatomic structures were rated positive for tumor infiltration, but diagnostic accuracy did not differ significantly in the subgroup of patients with histologic correlation from en bloc resection. CONCLUSION: Conventional 3-mm axial evaluation of head and neck MDCT proved to be sufficient in tumor staging. PMID- 18287454 TI - Diagnosis of blunt cerebrovascular injuries with 16-MDCT: accuracy of whole-body MDCT compared with neck MDCT angiography. AB - OBJECTIVE: The objective of our study was to determine whether whole-body 16-MDCT and neck MDCT angiography (MDCTA) can be used to diagnose blunt cerebrovascular injuries with comparable accuracy using angiography as the reference standard. MATERIALS AND METHODS: Retrospective review of radiology reports and prospective clinical observation identified 108 blunt trauma patients examined with either whole-body MDCT or neck MDCTA followed by angiography over a 23-month period. From this group, results from the retrospective interpretations of 77 whole-body MDCT and 48 neck MDCTA examinations were compared with the results extracted from angiography reports to estimate the accuracy of each protocol for detecting blunt cerebrovascular injuries. Fisher's exact test was used to determine any significant difference in the results of those patients scanned with both protocols. RESULTS: Angiography confirmed blunt cerebrovascular injury in 83 patients, with 25 (30%) showing multiple sites of injury. Most injuries were detected in cervical arterial segments. The respective sensitivities of whole body MDCT and neck MDCTA were 69% (36/52) and 64% (16/25) for cervical internal carotid artery injuries, and specificities were 82% (58/71) and 94% (49/52). Respective sensitivities for cervical vertebral artery injuries were 74% (17/23) and 68% (13/19), and specificities were 91% (60/66) and 100% (40/40). In 17 patients scanned with both protocols, the results were not significantly different (carotid arteries, p = 1.00; vertebral arteries, p = 0.68). CONCLUSION: Whole-body 16-MDCT and neck MDCTA can be used to diagnose blunt cerebrovascular injuries with comparable accuracy. Both show high specificities for cervical arterial injury. The sensitivity of whole-body 16-MDCT is sufficiently high to serve as an initial screening examination for blunt cerebrovascular injuries. PMID- 18287455 TI - Percutaneous ethanol injection of hyperfunctioning thyroid nodules: long-term follow-up in 125 patients. AB - OBJECTIVE: The purpose of this study was to assess the long-term efficacy of percutaneous ethanol injection (PEI) for the treatment of hyperfunctioning thyroid nodules. MATERIALS AND METHODS: One hundred twenty-five patients (88 women, 37 men; age range, 17-76 years; mean age, 53 years) with 127 hyperfunctioning thyroid nodules (volume, 1.2-90 mL; mean, 10.3 mL) were treated with PEI. There were 1-11 PEI sessions per patient (average, 3.9) performed, with injection of 1-14 mL of ethanol per session (total injected ethanol per patient, 3-108 mL; mean, 14.0 mL). Efficacy of the treatment was assessed with color Doppler sonography; scintigraphy; and free triiodothyronine (FT3), free thyroxine (FT4), and thyroid-stimulating hormone (TSH) assays. Follow-up (9-144 months; median, 60 months) was performed with TSH and color Doppler sonography every 2 months for 6 months and every 6 months thereafter. RESULTS: Three (2.4%) of 125 patients refused completion of PEI therapy because of pain. Results are reported in 122 patients with 124 nodules. All 122 patients showed posttreatment normal levels of FT3, FT4, and TSH. A complete cure (absent uptake in the nodule and recovery of normal uptake in the thyroid parenchyma) was obtained in 113 (93%) of 122 patients-115 (92.7%) of 124 treated nodules. Residual hyperfunctioning nodular tissue along with decreased thyroid parenchyma uptake (partial cure) was present in nine patients accounting for nine (7.3%) of 124 nodules. Rates of complete cure after PEI were: overall nodules, 115 (92.7%) of 124; nodules < or = 10 mL, 63 (94.0%) of 67; nodules > 10 to < or = 30 mL, 32 (91.4%) of 35; nodules > 30 to < or = 60 mL, 17 (89.5%) of 19; nodules > 60 mL, three (100%) of three. The overall rate of major complications (transient laryngeal nerve damage, two patients; abscess and hematoma, one patient each) was four (3.2%) of 125 patients. Follow-up examinations showed marked shrinkage of 112 treated nodules ranging from 50% to 90% of the pretreatment volume (mean, 66%) and new growth of hyperfunctioning tissue in four patients at color Doppler sonography and scintigraphy at 12, 18, 18, and 48 months' follow-up, respectively. However, all patients remained euthyroid (low or normal TSH and normal FT3 and FT4) during follow-up. CONCLUSION: PEI of hyperfunctioning thyroid nodules seems to be an effective and safe alternative to traditional treatment. It also appears to be effective in patients with hyperfunctioning thyroid nodules larger than 30 mL. PMID- 18287456 TI - Imaging characteristics of atypical teratoid-rhabdoid tumor in children compared with medulloblastoma. AB - OBJECTIVE: The purpose of our study was to compare the imaging characteristics of atypical teratoid-rhabdoid tumor with medulloblastoma and seek distinguishing features that can aid in preoperative diagnosis. MATERIALS AND METHODS: Preoperative MRI examinations of 55 patients (36 medulloblastomas and 19 atypical teratoid-rhabdoid tumors) were analyzed retrospectively. Imaging characteristics of atypical teratoid-rhabdoid tumor and medulloblastoma were assessed with conventional MRI and CT. Diffusion-weighted imaging (DWI) was available in 27 patients (19 medulloblastomas and eight atypical teratoid-rhabdoid tumors). Apparent diffusion coefficient (ADC) values were calculated for 14 medulloblastomas and six atypical teratoid-rhabdoid tumors. RESULTS: Both atypical teratoid-rhabdoid tumors in general and infratentorial atypical teratoid rhabdoid tumors presented at a younger age than medulloblastomas. Eleven of 19 atypical teratoid-rhabdoid tumors were infratentorial. Cerebellopontine angle (CPA) involvement was more frequent (8/11, 72.7%) in atypical teratoid-rhabdoid tumor than in medulloblastoma (4/36, 11.1%) (p < 0.001). Intratumoral hemorrhage was more common in atypical teratoid-rhabdoid tumor (9/19, 47.4%) than in medulloblastoma (2/36, 5.6%) (p < 0.0001). All atypical teratoid-rhabdoid tumors and all medulloblastomas for which DWI was available displayed increased signal intensity on DWI compared with normal brain parenchyma. The mean ADC values for tumor types were not significantly different. CONCLUSION: Atypical teratoid rhabdoid tumor presents at a younger age than medulloblastoma. Although atypical teratoid-rhabdoid tumor and medulloblastoma display similar imaging characteristics on conventional MRI, CPA involvement and intratumoral hemorrhage are more common in atypical teratoid-rhabdoid tumor. If a pediatric posterior fossa mass that displays restricted diffusion is involving the CPA, atypical teratoid-rhabdoid tumor is a more likely consideration than medulloblastoma. PMID- 18287457 TI - Clinical utility of hybrid SPECT-CT in endocrine neoplasia. AB - OBJECTIVE: Imaging of endocrine neoplasms often involves a combination of anatomic and functional techniques including sonography, CT, MRI, and scintigraphy. Recent technologic advances have enabled hybrid imaging using SPECT CT, which combines anatomic and functional techniques to allow accurate localization of tumors, increased detection of recurrent and metastatic disease, and exclusion of physiologic uptake. CONCLUSION: SPECT-CT provides improved specificity and diagnostic confidence helping to guide conventional management and assess suitability for targeted radionuclide therapy. PMID- 18287458 TI - Imaging features of sarcoidosis on MDCT, FDG PET, and PET/CT. AB - OBJECTIVE: The objectives of this article are to discuss the epidemiology and natural history of sarcoidosis; to review the classic imaging features of sarcoidosis on radiography, CT, and 67Ga nuclear medicine scans; and to present clinical examples of sarcoidosis as seen on PET and PET/CT in the chest, abdomen and pelvis, and bones. CONCLUSION: The imaging features of sarcoidosis are diverse and can be seen on a variety of imaging techniques. It is important for radiologists and nuclear medicine physicians to recognize the common imaging features and patterns of sarcoidosis in order to raise the possibility in the appropriate clinical setting. PMID- 18287459 TI - Radiological reasoning: miliary disease, vertebral osteomyelitis, and soft-tissue abscesses. AB - OBJECTIVE: We present a case of disseminated coccidioidomycosis with miliary disease and extrathoracic spread to the breast, the retroperitoneum, the soft tissues of the neck, and multiple vertebrae with spinal cord compression. We discuss the differential diagnosis of the imaging presentation, as well as the specific clinical and imaging features of coccidioidomycosis. CONCLUSION: Disseminated coccidioidomycosis in a nonendemic area can be difficult to diagnose, even with an excellent clinical history, as almost every organ system can be involved. Widespread disease can be seen and spinal involvement can easily be mistaken for malignancy, Pott's disease, or other granulomatous disease. Miliary disease with concomitant breast involvement is a rare presentation of disseminated disease. Detection of specific radiographic patterns of involvement and recognition of travel to or from an endemic area can lead to an accurate diagnosis and earlier treatment. PMID- 18287460 TI - Teaching insights from adult learning theory. AB - How do undergraduate and graduate students learn? How can we use what we know about the learning process to teach more effectively? While the research has yet to connect fundamental processes in the brain to what we do as teachers with certainty, the past half-century of work on adult learning has produced several useful theories that can inform instructional choices. This article provides an overview of three learning models that yield insights into teaching practice- novice/expert behaviors, cognitive development, and learning styles--along with ways in which instructors can draw on these models in course planning and classroom teaching. Application of the theories toward refinement, reduction, and replacement of live animals in the veterinary medical curriculum is also discussed. PMID- 18287461 TI - Teaching as a first, second, or third language. AB - The 2005 National Carl J. Norden/Pfizer Distinguished Teaching Award may bear my name, but clearly it is shared recognition for the institutions, numerous individuals, and varied professional experiences that inspired and influenced a veterinarian who became a teacher whom students seemed to value. My education, practice experience, exposure to some superb educators, and feedback from students have provided a diverse background of knowledge and experiences that have allowed me to adapt to teaching opportunities in courses other than clinical pathology and to develop my own style of instruction, teaching philosophy, and examination strategies and formats. In these times of limited funds and faculty recruitment, teaching programs have been developed to meet our learning objectives and make efficient use of monetary and faculty resources. The description that follows summarizes my thoughts and experiences. PMID- 18287462 TI - Education in veterinary pathology: innovations and global challenges. PMID- 18287463 TI - Veterinary pathology in the United Kingdom: past, present, and future. AB - This article presents a historical perspective on veterinary anatomic pathology in the United Kingdom from the late nineteenth century to the present. Prior to World War II, the specialty was a rather general one that also included bacteriology and parasitology and was only slightly affected by strong Germanic developments in cell and tissue pathology. The few notable figures of this era include John McFadyean, Sidney Gaiger, and J.R.M Innes. The specialty developed strongly in the second half of the twentieth century, led by a small number of individuals, and was greatly aided by the development of specialist colleges and residency training. Key individuals of this era include W.F. Blakemore, Ernest Cotchin, R.J.M. Franklin, W.F.H. Jarrett, A.R. Jennings, and A.C. Palmer. A remarkable feature of this period has been the increased employment of veterinary pathologists in biomedical industry and in private diagnostic laboratories. While standards of pathology practice have benefited from the college initiatives, there are major financial constraints on the availability of funded training posts in the United Kingdom, and there remain considerable shortages in the supply of pathologists trained to contemporary standards. The acknowledged professional and scientific importance of veterinary pathology needs to be translated into effective financial support for the training that underpins competence in this specialty. Further developments seem likely to be dominated by advances in the technology of tissue handling, applications of molecular biology to pathology, and greater use of telepathology in teaching, in quality assurance, and in continuing professional development. PMID- 18287464 TI - Pathology in the medical curriculum in the United Kingdom. AB - In the United Kingdom, the medical students of yesteryear undertook a detailed pathology course, often with its own major examinations. This contributed to the increasing overload of factual information that was common in medical curricula. Following General Medical Council reports on medical education, pathology is now more fully integrated into clinical work and into problem-based learning, albeit with greatly reduced contact time with pathologists. Teaching methods have shifted from didactic presentations, such as lectures, to more varied methods, with emphasis on self-access and interactivity. Technological advances have brought well-illustrated textbooks, computer-assisted learning programs, Web resources, and electronic communication. There has been a decline in the role of museum specimens, microscopy, and, partly following a worldwide trend and partly following adverse media publicity, the traditional autopsy. Assessment methods have evolved, with integration of pathology into wider examinations. The greater drive toward increasing funding through the Research Assessment Exercise has had an adverse effect on teaching throughout universities. Despite these setbacks, the importance of pathology as a clinical discipline will ensure its continued role in medical education. PMID- 18287465 TI - Integrated Case-Based Applied Pathology (ICAP): a diagnostic-approach model for the learning and teaching of veterinary pathology. AB - Integrative Case-Based Applied Pathology (ICAP) cases form one component of learning and understanding the role of pathology in the veterinary diagnostic process at the Faculty of Veterinary Science, University of Sydney. It is a strategy that focuses on student-centered learning in a problem-solving context in the year 3 curriculum. Learning exercises use real case material and are primarily delivered online, providing flexibility for students with differing learning needs, who are supported by online, peer, and tutor support. The strategy relies heavily on the integration of pre-clinical and para-clinical information with the introduction of clinical material for the purposes of a logical three-level, problem-oriented approach to the diagnosis of disease. The focus is on logical diagnostic problem solving, primarily using gross pathology and histopathological material, with the inclusion of microbiological, parasitological, and clinical pathological data. The ICAP approach is linked to and congruent with the problem-oriented approach adopted in veterinary medicine and the case-based format used by one of the authors (PJC) for the teaching and learning of veterinary clinical pathology in year 4. Additionally, final-year students have the opportunity, during a diagnostic pathology rotation, to assist in the development and refinement of further ICAPs, which reinforces the importance of pathology in the veterinary diagnostic process. Evidence of the impact of the ICAP approach, based primarily on student surveys and staff peer feedback collected over five years, shows that discipline-specific learning, vertical and horizontal integration, alignment of learning outcomes and assessment, and both veterinary and generic graduate attributes were enhanced. Areas for improvement were identified in the approach, most specifically related to assistance in the development of generic teamwork skills. PMID- 18287466 TI - An integrative and case-based approach to the teaching of general and systemic pathology. AB - Since 1993, the pre-clinical phase of the professional (DVM) curriculum at Michigan State University College of Veterinary Medicine has included two pathology courses in which both anatomic and clinical pathologists collaborate to teach concepts in general and systemic pathology. Topics such as inflammation, circulatory disturbances, and neoplasia are taught in this collaborative manner in the year 1 General Pathology course, and pathology of the digestive system (including liver and pancreas), urinary system, and lymphoid system are "team taught" in the year 2 Clinical and Systemic Pathology course. We feel that this approach gives students an appreciation of the whole-body dynamics of a disease process as it occurs in bone marrow, peripheral blood, body fluids, and tissues and that it leads to a deep understanding of pathologic processes. In addition, the use of "active learning" instructional strategies grounded in case discussions further enhances students' understanding of important concepts by demonstration of practical applications and serves to generate strong interest in learning the subject matter. Integration of concepts of pathology with those taught concurrently in other courses, such as those in physiology and microbiology, is also an important component of pathology instruction in the pre clinical curriculum. PMID- 18287467 TI - Virtual microscopy in a veterinary curriculum. AB - Teaching faculty in the University of Tennessee College of Veterinary Medicine assist students in their professional education by providing a new way of viewing microscopic slides digitally. Faculty who teach classes in which glass slides are used participate in a program called Virtual Microscopy. Glass slides are digitized using a state-of-the-art integrated system, and a personal computer functions as the "microscope." Additionally, distribution of the interactive images is enhanced because they are available to students online. The digital slide offers equivalent quality and resolution to the original glass slide viewed on a microscope and has several additional advantages over microscopes. Students can choose to examine the entire slide at any of several objectives; they are able to access the slides (called WebSlides) from the college's server, using either Internet Explorer or a special browser developed by Bacus Laboratories, Inc.,(a) called the WebSlide browser, which lets the student simultaneously view a low-objective image and one or two high-objective images of the same slide. The student can "move the slide" by clicking and dragging the image to a new location. Easy archiving, annotation of images, and Web conferencing are additional features of the system. PMID- 18287468 TI - Evolution of a course in veterinary clinical pathology: the application of case based writing assignments to focus on skill development and facilitation of learning. AB - RATIONALE FOR STUDY: To encourage application and critical thinking, case-based writing assignments and grading rubrics were developed for use in a second-year core clinical pathology course. Objectives were to describe how this teaching technique was adapted to a large-class setting and how student perceptions of the learning experience guided modifications of this teaching technique over two presentations of the course and plans for a third presentation. Our goal was to enhance learning by encouraging application of course material to clinical situations and thereby improve students' ability to organize and communicate information. Furthermore, we evaluated the influence of instructor feedback on the learning process. METHODS: With assistance from the University of Minnesota Center for Writing, assignments and grading rubrics were developed. Students completed course evaluation surveys designed to elicit feedback on the impact of the assignments. RESULTS: Increased learner confidence was reflected in larger self-reported increases in understanding of the material and ability to apply information and in increased feelings of preparedness for class and examinations. A large majority of students advocated the use of such assignments in the course in future years, and modifications to make grading and evaluation of assignments more efficient are underway. CONCLUSIONS: Investment of faculty and student time in case-based writing assignments in the veterinary clinical pathology curriculum appears to increase student engagement with material and learner confidence. Future studies should address the impact of this type of assignment more specifically on clinical reasoning and communication skills and on long-term retention of material. PMID- 18287469 TI - Teaching medical pathology in the twenty-first century: virtual microscopy applications. AB - Virtual microscopy (VM) has been implemented and evaluated in the histology and general and systemic pathology courses at the University of Iowa Carver College of Medicine. Advantages of VM over traditional microscopy include accessibility and efficiency of learning and the ability to integrate VM with computer-assisted interactive learning. Advantages of using VM as opposed to digital photomicrographs include the ability to pan and zoom, explore the slide, and make independent observations. Although VM is used in a case-based format for teaching histopathology to medical students at the University of Iowa, VM may also be effectively implemented in other medical-student teaching models, including integrated and problem-based learning curricula and the classical pathology laboratory. Additional Iowa venues and courses using VM teaching include pathology of human disease for bioscience graduate students, cytology education, a comparative pathology research resource, and histology and histopathology for veterinary medicine. This article reviews the history and evolution of VM in medical pathology and its implementation at Iowa. PMID- 18287470 TI - Introduction and evaluation of virtual microscopy in teaching veterinary cytopathology. AB - Virtual microscopy (VM) uses a computer to view digitized slides and is comparable to using a microscope to view glass slides. This technology has been assessed in human medical education for teaching histology and histopathology, but, to the authors' knowledge, no one has evaluated its use in teaching cytopathology in veterinary medical education. We hypothesize that students will respond positively to the use of VM for viewing cytopathology preparations and that the technology can be successfully used for student assessment. To test this hypothesis, we surveyed students regarding their level of satisfaction with features of the VM system, their preference for use of VM in the curriculum, and the potential influence virtual slides may have on student study habits; student performance on a traditional cytopathology practical examination and a similar exam using VM was evaluated. Our results show that student perception of the VM system is generally very positive, with some concerns about resolution and the need for continued exposure to traditional microscopy. Within the curriculum, students indicated a preference for the option of using virtual slides for studying and take-home exercises. Overwhelmingly, students wanted either hybrid laboratory sessions or sessions using glass slides with virtual slides available for study and review. Students identified many VM test-taking features as advantageous compared with traditional glass-slide practical exams as traditionally administered. However, students indicated a strong preference for continued use of traditional microscopy for graded practical exams. Students may be more likely to study slides in preparation for practical examinations if virtual slides are available. Results also indicate that VM can be used successfully for assessment purposes, but students should receive training in using virtual slides if the technology will be used for assessment. PMID- 18287471 TI - Introduction to the Swiss way of teaching veterinary pathology in the twenty first century: application of e-learning modules. AB - The teaching of pathology within the veterinary medical curriculum extends through the entire training period and has a dual role. General pathology deals with principles of disease processes as a basis for understanding the reactions of a multi-cellular organism to adverse effects; organ pathology builds on the principles of general pathology and explains the malfunctions of individual organs. Pathology is heavily image dependent and best taught in a highly interactive manner. The Institute of Veterinary Pathology, University of Zurich (IVPZ), has been actively developing concepts for teaching pathology in the new veterinary curriculum, which demands more active participation from students, with 20% of their study time devoted to individual study using traditional materials (books, articles, etc.) and e-learning modules. The Swiss Virtual Animal Pathologist is designed to cover and support the central elements of the Veterinary Pathology curriculum of the Vetsuisse Faculty. The needs of students and staff of the participating institutions for an interactive learning platform to supplement the existing face-to-face lectures and tutorials are the highest priority of this initiative. PMID- 18287472 TI - Future directions in training veterinarians for careers in toxicological pathology in the United Kingdom. AB - There is currently a global shortage of veterinary pathologists in all sectors of the discipline, and recruitment of toxicological pathologists is a particular problem for the pharmaceutical industry. Efforts to encourage veterinarians to consider alternative career paths to general practice must start at the undergraduate level, with provision of structured career guidance and strong role models from pathology and research disciplines. It is also imperative that both the importance of biomedical research and the role of animal models be clearly understood by both university staff and undergraduates. Traditionally, much post graduate training in toxicological pathology is done "on the job" in the United Kingdom, but completion of a residency and/or PhD program is recognized as a good foundation for a career in industry and for successful completion of professional pathology examinations. New models of residency training in veterinary pathology must be considered in the United Kingdom to enable a more tailored approach to training toward specific career goals. A modular approach to residency training would allow core skills to be maintained, while additional training would target specific training requirements in toxicological pathology. Exposure to laboratory animal pathology, toxicology, research methodology, and management skills would all be of benefit as an introduction to a career in toxicological pathology. However, long-term funding for UK residencies remains a problem that must be resolved if future recruitment needs in veterinary pathology are to be met. PMID- 18287473 TI - North American veterinary pathology residency training programs: an overview of where they are today and where they were five years ago, with an analysis of trends. AB - An e-mail/telephone survey of all active North American residency training programs in veterinary pathology was conducted in September 2005. The purpose of this survey was to determine current numbers of trainees, their program length and type, and salaries; to compare current numbers to five years earlier; and, finally, to gauge interest in expanding current programs. All 41 training institutions contacted responded to the survey. Briefly, the survey found that there are currently 235 veterinary pathology residents, for a mean of 5.7 residents per training program. The number of residents currently in training programs and the number of applicants for these programs has increased compared to five years earlier. There is widespread interest in further expanding capacity in these programs, and the coalition of the American College of Veterinary Pathologists and the Society of Toxicologic Pathology is a well-known source of possible funding for additional residents. This survey report further documents the numbers of combined residency/PhD programs, average starting salaries for new residents, outside sponsorship effects on pathology training programs, and some of the common concerns regarding veterinary pathology training programs voiced by the respondents. While residency training capacity has expanded in the last five years, and there is widespread desire to further expand these training programs, a shortage of veterinary pathologists for future market needs will need to be addressed by increased funding from as yet unspecified sources. PMID- 18287474 TI - Developing and fostering a dynamic program for training in veterinary pathology and clinical pathology: veterinary students to post-graduate education. AB - Recent reports project a deficiency of veterinary pathologists, indicating a need to train highly qualified veterinary pathologists, particularly in academic veterinary medicine. The need to provide high-quality research training for veterinary pathologists has been recognized by the veterinary pathology training program of the Ohio State University (OSU) since its inception. The OSU program incorporates elements of both residency training and graduate education into a unified program. This review illustrates the components and structure of the training program and reflects on future challenges in training veterinary pathologists. Key elements of the OSU program include an experienced faculty, dedicated staff, and high-quality students who have a sense of common mission. The program is supported through cultural and infrastructure support. Financial compensation, limited research funding, and attractive work environments, including work-life balance, will undoubtedly continue to be forces in the marketplace for veterinary pathologists. To remain competitive and to expand the ability to train veterinary pathologists with research skills, programs must support strong faculty members, provide appropriate infrastructure support, and seek active partnerships with private industry to expand program opportunities. Shortages of trained faculty may be partially resolved by regional cooperation to share faculty expertise or through the use of communications technology to bridge distances between programs. To foster continued interest in academic careers, training programs will need to continue to evolve and respond to trainees' needs while maintaining strong allegiances to high-quality pathology training. Work life balance, collegial environments that foster a culture of respect for veterinary pathology, and continued efforts to reach out to veterinary students to provide opportunities to learn about the diverse careers offered in veterinary pathology will pay long-term dividends for the future of the profession. PMID- 18287475 TI - The European College of Veterinary Pathologists (ECVP): the professional body for European veterinary pathologists. AB - The European College of Veterinary Pathologists (ECVP) was established in 1995 with the aim of advancing veterinary pathology and promoting high standards within the specialty in Europe. The ECVP is one of 21 European colleges recognized by the European Board of Veterinary Specialisation (EBVS), which represents a quality-assurance system for European veterinary specialists. Until the ECVP was founded, there was no unified European system recognizing the specialty of pathology, and many European countries followed their own qualification systems, which varied in form and standard. The ECVP provides an annual certifying examination, the passing of which is required to gain membership (diplomate status) in the college. This qualification is now accepted on equal terms by the well-established American College of Veterinary Pathologists (ACVP). In line with EBVS requirements, the ECVP has also established a standard continuing professional development (CPD) and re registration system for its membership. Furthermore, it has promoted and unified European post-graduate training in veterinary pathology by setting up requirements for residency training programs and making registration and monitoring of these programs by the ECVP a prerequisite for approval of an institution as a training facility. The concurrent establishment, together with the European Society of Veterinary Pathology, of an annual summer school that trains residents for the certifying examination has further fostered European post-graduate training. Within 10 years, the ECVP has succeeded in establishing common standards and a unified approach to veterinary pathology throughout Europe. This article describes the evolution and organization of the ECVP. PMID- 18287476 TI - Residency training programs in veterinary clinical pathology: a comparison of experiences at two institutions. AB - Two institutions with different residency training formats in clinical pathology are compared with respect to application procedures, learning and teaching opportunities, learning resources, research training, publication requirements, and assessment methods of the program and trainees. The University of Florida and Purdue University programs are both based on an emphasis in morphologic recognition and interpretation of disease processes as well as training in basic science and applied research principles. The progress of trainees through each program is carefully monitored to meet individual needs as well as to meet the training requirements to allow candidates to sit for the certifying examination in clinical pathology. Periodic mock board exams are a critical tool to assess trainee progress and learning. The differences in format focus on coursework and publication requirements as well as on program assessment tools. While one program provides training in the form of 75% clinical diagnostic service, the other uses a mixture of 50% coursework and 50% clinical diagnostic training. Despite the contrast between a pure residency training program and one combining residency training with an MS degree, both institutions provide a solid program structure, ample learning resources, and adequate faculty mentorship to produce a high pass rate of board-certified specialists, the major focus for both programs. Numbers of post-training employment positions for both institutions are similar for those selecting faculty positions at veterinary schools. During the period studied, however, the combined residency and MS graduate program at Purdue University produced more graduates employed in pharmaceutical and biotechnology companies, while the residency program at the University of Florida produced more graduates employed by diagnostic laboratories. PMID- 18287477 TI - The ECVP/ESVP summer school in veterinary pathology: high-standard, structured training for young veterinary pathologists. AB - This article describes the ECVP/ESVP Summer School in Veterinary Pathology, a new annual two-week European training facility established by the European College of Veterinary Pathologists (ECVP) in collaboration with the European Society of Veterinary Pathology (ESVP). The aim of the Summer Schools is to provide Europe wide, harmonized, top-standard theoretical and practical post-graduate training for veterinarians specializing in veterinary pathology. In particular, it aims to support trainees in veterinary pathology in their individual preparation for the ECVP certifying examination. Ultimately, it aims to provide young pathologists with the skills and knowledge necessary to participate in international, high quality research and the tools for applying international standards to their own research and for independent study for the ECVP certifying examination, even if they do not work in comparable academic environments and do not have the same level of local support and training. The ECVP/ESVP Summer Schools take place in European countries, with local organization from a university department of veterinary pathology. Each event comprises modules provided by internationally recognized specialists in their specific fields of expertise on different organ systems, diseases of specific species, specific techniques, and specific topics relevant to pathology, forming a cycle of four events to cover all major topics. Every two years a mock exam is organized as a tool to monitor individual progress in preparing for the ECVP certifying examination. PMID- 18287478 TI - New approaches for education and training in veterinary public health: the SAPUVET projects. AB - Continued contact between humans and animals, in combination with the ever increasing movement of human and animal populations that is one effect of globalization, contributes to the spread of diseases, often with detrimental effects on public health. This has led professionals involved in both animal health and public health to recognize veterinary public health (VPH) as a key area for their activities to address the human-animal interface. Veterinarians, a profession with major involvement in this field, are in need of specific knowledge and skills to prevent and control public-health problems. As a result, VPH must be directly integrated into veterinary educational programs. At present, only few veterinary schools have specific VPH programs; in most institutions, VPH does not feature as a specific subject in either undergraduate or post-graduate curricula. SAPUVET and SAPUVETNET II are network projects supported by the ALFA program of the European Union (EU). Their main objectives are to reach a common understanding between European and Latin American universities in the definition of the areas in which VPH is important in their respective countries, and to design a harmonized training program for veterinarians in VPH, by making use of new technological applications and innovative teaching methodologies. The elaboration of educational material in combination with case studies presenting real-life problems provides a basis to apply the knowledge acquired on VPH. It is envisaged that the material and modules developed during the two projects will be integrated into the veterinary curricula of the participating universities, as well as in other partner organizations. PMID- 18287479 TI - Interactive computerized learning program exposes veterinary students to challenging international animal-health problems. AB - This article describes a computerized case-based CD-ROM (CD) on international animal health that was developed to give veterinary students an opportunity to "virtually" work alongside veterinarians and other veterinary students as they try to solve challenging disease problems relating to tuberculosis in South African wildlife, bovine abortion in Mexico, and neurologic disease in horses in Rapa Nui, Chile. Each of the three case modules presents, in a highly interactive format, a problem or mystery that must be solved by the learner. As well as acquiring information via video clips and text about the specific health problem, learners obtain information about the different countries, animal-management practices, diagnostic methods, related disease-control issues, economic factors, and the opinions of local experts. After assimilating this information, the learner must define the problem and formulate an action plan or make a recommendation or diagnosis. The computerized program invokes three principles of adult education: active learning, learner-centered education, and experiential learning. A medium that invokes these principles is a potentially efficient learning tool and template for developing other case-based problem-solving computerized programs. The program is accessible on the World Wide Web at . A broadband Internet connection is recommended, since the modules make extensive use of embedded video and audio clips. Information on how to obtain the CD is also provided. PMID- 18287480 TI - Virtual international experiences in veterinary medicine: an evaluation of students' attitudes toward computer-based learning. AB - While many studies have evaluated whether or not factual information can be effectively communicated using computer-aided tools, none has focused on establishing and changing students' attitudes toward international animal-health issues. The study reported here was designed to assess whether educational modules on an interactive computer CD elicited a change in veterinary students' interest in and attitudes toward international animal-health issues. Volunteer veterinary students at seven universities (first-year students at three universities, second-year at one, third-year at one, and fourth-year at two) were given by random assignment either an International Animal Health (IAH) CD or a control CD, ParasitoLog (PL). Participants completed a pre-CD survey to establish baseline information on interest and attitudes toward both computers and international animal-health issues. Four weeks later, a post-CD questionnaire was distributed. On the initial survey, most students expressed an interest in working in the field of veterinary medicine in another country. Responses to the three pre-CD questions relating to attitudes toward the globalization of veterinary medicine, interest in foreign animal disease, and inclusion of a core course on international health issues in the veterinary curriculum were all positive, with average values above 3 (on a five-point scale where 5 represented strong agreement or interest). Almost all students considered it beneficial to learn about animal-health issues in other countries. After students reviewed the IAH CD, we found a decrease at four universities, an increase at one university, and no change at the remaining two universities in students' interest in working in some area of international veterinary medicine. However, none of the differences was statistically significant. PMID- 18287481 TI - OLIVER: an online library of images for veterinary education and research. AB - As part of a strategic move by the University of Sydney toward increased flexibility in learning, the Faculty of Veterinary Science undertook a number of developments involving Web-based teaching and assessment. OLIVER underpins them by providing a rich, durable repository for learning objects. To integrate Web based learning, case studies, and didactic presentations for veterinary and animal science students, we established an online library of images and other learning objects for use by academics in the Faculties of Veterinary Science and Agriculture. The objectives of OLIVER were to maximize the use of the faculty's teaching resources by providing a stable archiving facility for graphic images and other multimedia learning objects that allows flexible and precise searching, integrating indexing standards, thesauri, pull-down lists of preferred terms, and linking of objects within cases. OLIVER offers a portable and expandable Web based shell that facilitates ongoing storage of learning objects in a range of media. Learning objects can be downloaded in common, standardized formats so that they can be easily imported for use in a range of applications, including Microsoft PowerPoint, WebCT, and Microsoft Word. OLIVER now contains more than 9,000 images relating to many facets of veterinary science; these are annotated and supported by search engines that allow rapid access to both images and relevant information. The Web site is easily updated and adapted as required. PMID- 18287482 TI - Unique educational methods to improve the veterinary employment selection process for rural mixed-animal practices. AB - The rural mixed-animal veterinarian is a critical control point for safe, wholesome, affordable food production and security. The population of students entering food-animal practice is decreasing, and future shortages are likely. Veterinary practice owners will continue to struggle to find associates to fill open positions. Identifying and hiring the correct veterinarian for an open position is a challenging proposition for the rural practitioner. Kansas State University hosted a forum to facilitate the hiring process and provide education regarding the mechanism of an effective selection interview. A unique experiential technique known as "speed interviews" was used to facilitate communication between conference participants and to practice newly acquired skills. A survey of participants revealed similar viewpoints toward most job attributes. Veterinary students and prospective employers expressed realistic expectations of job requirements, salaries, and debt load. Students expressed willingness to work and desire to practice in the types of practices defined by the veterinarians. The symposium provided valuable insight for practitioners and students regarding the recruitment process. Appropriate and accurate representation at the time of job/associate selection is critical for long-term success and employee retention. The goal of the event was to provide a service to both prospective employers and students by offering education regarding the employment selection interview process and placing attendees in an environment rich with people who have complimentary goals. PMID- 18287483 TI - Teaching veterinary students beef production medicine with student/producer teams. AB - Veterinary colleges face difficulties in meeting the demand for rural veterinarians with the scope to practice quality production medicine. Increasing population density around veterinary colleges, retaining the interest of students with a background in animal agriculture, and educating students without a farm background requires that veterinary colleges consider innovative ways to not only teach traditional food-animal practice but give future veterinarians the advanced skills the food industry demands. This article describes a three-year elective program, Beef Records Analysis, in which beef production medicine is taught by teaming a student and a beef producer together early in the student's veterinary education. These producer/student teams complete risk assessments, balance rations, collect financial and production information, and evaluate back grounding and feedlot enterprises. Students learn how to evaluate their producers using industry benchmarks and past performance records and how to communicate their findings back to their producer. Producers often make management decisions based on the students' findings, and, because the students maintain their relationships with producers for three years, they can assess the outcomes of the producers who follow or ignore their recommendations and interventions. Students share recommendations and outcomes associated with their herd with the entire class. This allows students to learn how to establish best management practices through objective analysis of outcomes of recommended practices of all herds represented in the class. While a formal assessment of the course is needed, the students rate the program very high on evaluations. PMID- 18287484 TI - The critical need for federal legislation to provide for the expansion of the veterinary workforce. AB - American veterinary medical education stands at a crucial point. To maintain the status quo and to meet ever-increasing societal needs within the United States and globally, it is essential that the veterinary medical profession expand its horizons and capabilities. If it does not, it will lose its current status. The profession faces a crucial shortage in the workforce it needs both to continue to perform its current functions and, more importantly, to meet its growing responsibilities. This is not a new situation, and the profession can learn from its past successes and failures. Action is necessary. The profession has the capability and expertise to meet these challenges, but it must activate these skills in order to succeed. PMID- 18287485 TI - Control of compound leaf development by FLORICAULA/LEAFY ortholog SINGLE LEAFLET1 in Medicago truncatula. AB - Molecular genetic studies suggest that FLORICAULA (FLO)/LEAFY (LFY) orthologs function to control compound leaf development in some legume species. However, loss-of-function mutations in the FLO/LFY orthologs result in reduction of leaf complexity to different degrees in Pisum sativum and Lotus japonicus. To further understand the role of FLO/LFY orthologs in compound leaf development in legumes, we studied compound leaf developmental processes and characterized a leaf development mutant, single leaflet1 (sgl1), from the model legume Medicago truncatula. The sgl1 mutants exhibited strong defects in compound leaf development; all adult leaves in sgl1 mutants are simple due to failure in initiating lateral leaflet primordia. In addition, the sgl1 mutants are also defective in floral development, producing inflorescence-like structures. Molecular cloning of SGL1 revealed that it encodes the M. truncatula FLO/LFY ortholog. When properly expressed, LFY rescued both floral and compound leaf defects of sgl1 mutants, indicating that LFY can functionally substitute SGL1 in compound leaf and floral organ development in M. truncatula. We show that SGL1 and LFY differed in their promoter activities. Although the SGL1 genomic sequence completely rescued floral defects of lfy mutants, it failed to alter the simple leaf structure of the Arabidopsis thaliana plants. Collectively, our data strongly suggest that initiation of lateral leaflet primordia required for compound leaf development involves regulatory processes mediated by the SGL1 function in M. truncatula. PMID- 18287486 TI - Examining the specific contributions of individual Arabidopsis metallothioneins to copper distribution and metal tolerance. AB - Metallothioneins (MTs) are small cysteine-rich proteins found in various eukaryotes. Plant MTs are classified into four types based on the arrangement of cysteine residues. To determine whether all four types of plant MTs function as metal chelators, six Arabidopsis (Arabidopsis thaliana) MTs (MT1a, MT2a, MT2b, MT3, MT4a, and MT4b) were expressed in the copper (Cu)- and zinc (Zn)-sensitive yeast mutants, Deltacup1 and Deltazrc1 Deltacot1, respectively. All four types of Arabidopsis MTs provided similar levels of Cu tolerance and accumulation to the Deltacup1 mutant. The type-4 MTs (MT4a and MT4b) conferred greater Zn tolerance and higher accumulation of Zn than other MTs to the Deltazrc1 Deltacot1 mutant. To examine the functions of MTs in plants, we studied Arabidopsis plants that lack MT1a and MT2b, two MTs that are expressed in phloem. The lack of MT1a, but not MT2b, led to a 30% decrease in Cu accumulation in roots of plants exposed to 30 mum CuSO(4). Ectopic expression of MT1a RNA in the mt1a-2 mt2b-1 mutant restored Cu accumulation in roots. The mt1a-2 mt2b-1 mutant had normal metal tolerance. However, when MT deficiency was combined with phytochelatin deficiency, growth of the mt1a-2 mt2b-1 cad1-3 triple mutant was more sensitive to Cu and cadmium compared to the cad1-3 mutant. Together these results provide direct evidence for functional contributions of MTs to plant metal homeostasis. MT1a, in particular, plays a role in Cu homeostasis in the roots under elevated Cu. Moreover, MTs and phytochelatins function cooperatively to protect plants from Cu and cadmium toxicity. PMID- 18287487 TI - Systemic signaling of the plant nitrogen status triggers specific transcriptome responses depending on the nitrogen source in Medicago truncatula. AB - Legumes can acquire nitrogen (N) from NO(3)(-), NH(4)(+), and N(2) (through symbiosis with Rhizobium bacteria); however, the mechanisms by which uptake and assimilation of these N forms are coordinately regulated to match the N demand of the plant are currently unknown. Here, we find by use of the split-root approach in Medicago truncatula plants that NO(3)(-) uptake, NH(4)(+) uptake, and N(2) fixation are under general control by systemic signaling of plant N status. Indeed, irrespective of the nature of the N source, N acquisition by one side of the root system is repressed by high N supply to the other side. Transcriptome analysis facilitated the identification of over 3,000 genes that were regulated by systemic signaling of the plant N status. However, detailed scrutiny of the data revealed that the observation of differential gene expression was highly dependent on the N source. Localized N starvation results, in the unstarved roots of the same plant, in a strong compensatory up-regulation of NO(3)(-) uptake but not of either NH(4)(+) uptake or N(2) fixation. This indicates that the three N acquisition pathways do not always respond similarly to a change in plant N status. When taken together, these data indicate that although systemic signals of N status control root N acquisition, the regulatory gene networks targeted by these signals, as well as the functional response of the N acquisition systems, are predominantly determined by the nature of the N source. PMID- 18287489 TI - Protein diffusion and macromolecular crowding in thylakoid membranes. AB - The photosynthetic light reactions of green plants are mediated by chlorophyll binding protein complexes located in the thylakoid membranes within the chloroplasts. Thylakoid membranes have a complex structure, with lateral segregation of protein complexes into distinct membrane regions known as the grana and the stroma lamellae. It has long been clear that some protein complexes can diffuse between the grana and the stroma lamellae, and that this movement is important for processes including membrane biogenesis, regulation of light harvesting, and turnover and repair of the photosynthetic complexes. In the grana membranes, diffusion may be problematic because the protein complexes are very densely packed (approximately 75% area occupation) and semicrystalline protein arrays are often observed. To date, direct measurements of protein diffusion in green plant thylakoids have been lacking. We have developed a form of fluorescence recovery after photobleaching that allows direct measurement of the diffusion of chlorophyll-protein complexes in isolated grana membranes from Spinacia oleracea. We show that about 75% of fluorophores are immobile within our measuring period of a few minutes. We suggest that this immobility is due to a protein network covering a whole grana disc. However, the remaining fraction is surprisingly mobile (diffusion coefficient 4.6 +/- 0.4 x 10(-11) cm(2) s(-1)), which suggests that it is associated with mobile proteins that exchange between the grana and stroma lamellae within a few seconds. Manipulation of the protein lipid ratio and the ionic strength of the buffer reveals the roles of macromolecular crowding and protein-protein interactions in restricting the mobility of grana proteins. PMID- 18287488 TI - Genetic dissection of hormonal responses in the roots of Arabidopsis grown under continuous mechanical impedance. AB - We investigated the role of ethylene and auxin in regulating the growth and morphology of roots during mechanical impedance by developing a new growing system and using the model plant Arabidopsis (Arabidopsis thaliana). The Arabidopsis seedlings grown horizontally on a dialysis membrane-covered agar plate encountered adequate mechanical impedance as the roots showed characteristic ethylene phenotypes: 2-fold reduction in root growth, increase in root diameter, decrease in cell elongation, and ectopic root hair formation. The root phenotype characterization of various mutants having altered response to ethylene biosynthesis or signaling, the effect of ethylene inhibitors on mechanically impeded roots, and transcription profiling of the ethylene responsive genes led us to conclude that enhanced ethylene response plays a primary role in changing root morphology and development during mechanical impedance. Further, the differential sensitivity of horizontally and vertically grown roots toward exogenous ethylene suggested that ethylene signaling plays a critical role in enhancing the ethylene response. We subsequently demonstrated that the enhanced ethylene response also affects the auxin response in roots. Taken together, our results provide a new insight into the role of ethylene in changing root morphology during mechanical impedance. PMID- 18287490 TI - DNA-binding study identifies C-box and hybrid C/G-box or C/A-box motifs as high affinity binding sites for STF1 and LONG HYPOCOTYL5 proteins. AB - LONG HYPOCOTYL5 (HY5) is a bZIP (basic leucine zipper) transcription factor that activates photomorphogenesis and root development in Arabidopsis (Arabidopsis thaliana). Previously, STF1 (soybean [Glycine max] TGACG-motif binding factor 1), a homologous legume protein with a RING-finger motif and a bZIP domain, was reported in soybean. To investigate the role of STF1, the phenotypes of transgenic Arabidopsis plants overexpressing STF1 and HY5 were compared. In addition, the DNA-binding properties of STF1 and HY5 were extensively studied using random binding site selection and electrophoretic mobility shift assay. Overexpression of STF1 in the hy5 mutant of Arabidopsis restored wild-type photomorphogenic and root development phenotypes of short hypocotyl, accumulation of chlorophyll, and root gravitropism with partial restoration of anthocyanin accumulation. This supports that STF1 is a homolog of HY5 with a role in light and hormone signaling. The DNA-binding properties of STF1 and HY5 are shown to be similar to each other in recognizing many ACGT-containing elements with a consensus sequence motif of 5'-(G/A)(G/A) TGACGT(C/G/A)(A/T/G)-3'. The motif represents a characteristically strong preference for flanking sequence to TGACGT and a larger sequence than the sequences recognized by the G-box binding factor and TGA protein families. The finding of C-box, hybrid C/G-, and C/A-boxes as high-affinity binding sites over the G-box and parameters associated with HY5 recognition define the criteria of HY5/STF1 protein-DNA interaction in the promoter regions. This study helps to predict the precise in vivo binding sites of the HY5 protein from the vast number of putative HY5 genomic binding sites analyzed by chromatin immunoprecipitation on chip. PMID- 18287491 TI - Transcriptional and metabolic adjustments in ADP-glucose pyrophosphorylase deficient bt2 maize kernels. AB - During the cloning of monogenic recessive mutations responsible for a defective kernel phenotype in a Mutator-induced Zea mays mutant collection, we isolated a new mutant allele in Brittle2 (Bt2), which codes for the small subunit of ADP glucose pyrophosphorylase (AGPase), a key enzyme in starch synthesis. Reverse transcription-polymerase chain reaction experiments with gene-specific primers confirmed a predominant expression of Bt2 in endosperm, of Agpsemzm in embryo, and of Agpslzm in leaf, but also revealed considerable additional expression in various tissues for all three genes. Bt2a, the classical transcript coding for a cytoplasmic isoform, was almost exclusively expressed in the developing endosperm, whereas Bt2b, an alternative transcript coding for a plastidial isoform, was expressed in almost all tissues tested with a pattern very similar to that of Agpslzm. The phenotypic analysis showed that, at 30 d after pollination (DAP), mutant kernels were plumper than wild-type kernels, that the onset of kernel collapse took place between 31 and 35 DAP, and that the number of starch grains was greatly reduced in the mutant endosperm but not the mutant embryo. A comparative transcriptome analysis of wild-type and bt2-H2328 kernels at middevelopment (35 DAP) with the 18K GeneChip Maize Genome Array led to the conclusion that the lack of Bt2-encoded AGPase triggers large-scale changes on the transcriptional level that concern mainly genes involved in carbohydrate or amino acid metabolic pathways. Principal component analysis of (1)H nuclear magnetic resonance metabolic profiles confirmed the impact of the bt2-H2328 mutation on these pathways and revealed that the bt2-H2328 mutation did not only affect the endosperm, but also the embryo at the metabolic level. These data suggest that, in the bt2-H2328 endosperms, regulatory networks are activated that redirect excess carbon into alternative biosynthetic pathways (amino acid synthesis) or into other tissues (embryo). PMID- 18287492 TI - Advanced data-mining strategies for the analysis of direct-infusion ion trap mass spectrometry data from the association of perennial ryegrass with its endophytic fungus, Neotyphodium lolii. AB - Direct-infusion mass spectrometry (MS) was applied to study the metabolic effects of the symbiosis between the endophytic fungus Neotyphodium lolii and its host perennial ryegrass (Lolium perenne) in three different tissues (immature leaf, blade, and sheath). Unbiased direct-infusion MS using a linear ion trap mass spectrometer allowed metabolic effects to be determined free of any preconceptions and in a high-throughput fashion. Not only the full MS(1) mass spectra (range 150-1,000 mass-to-charge ratio) were obtained but also MS(2) and MS(3) product ion spectra were collected on the most intense MS(1) ions as described previously (Koulman et al., 2007b). We developed a novel computational methodology to take advantage of the MS(2) product ion spectra collected. Several heterogeneous MS(1) bins (different MS(2) spectra from the same nominal MS(1)) were identified with this method. Exploratory data analysis approaches were also developed to investigate how the metabolome differs in perennial ryegrass infected with N. lolii in comparison to uninfected perennial ryegrass. As well as some known fungal metabolites like peramine and mannitol, several novel metabolites involved in the symbiosis, including putative cyclic oligopeptides, were identified. Correlation network analysis revealed a group of structurally related oligosaccharides, which differed significantly in concentration in perennial ryegrass sheaths due to endophyte infection. This study demonstrates the potential of the combination of unbiased metabolite profiling using ion trap MS and advanced data-mining strategies for discovering unexpected perturbations of the metabolome, and generating new scientific questions for more detailed investigations in the future. PMID- 18287493 TI - Elongation changes of exploratory and root hair systems induced by aminocyclopropane carboxylic acid and aminoethoxyvinylglycine affect nitrate uptake and BnNrt2.1 and BnNrt1.1 transporter gene expression in oilseed rape. AB - Ethylene is a plant hormone that plays a major role in the elongation of both exploratory and root hair systems. Here, we demonstrate in Brassica napus seedlings that treatments with the ethylene precursor, aminocyclopropane carboxylic acid (ACC) and the ethylene biosynthesis inhibitor, aminoethoxyvinylglycine (AVG), cause modification of the dynamic processes of primary root and root hair elongation in a dose-dependent way. Moreover, restoration of root elongation in AVG-treated seedlings by 1 mm l-glutamate suggested that high concentrations of AVG affect root elongation through nonoverlapping ethylene metabolic pathway involving pyridoxal 5'-P-dependent enzymes of nitrate (N) metabolism. In this respect, treatments with high concentrations of ACC and AVG (10 mum) over 5 d revealed significant differences in relationships between root growth architecture and N uptake capacities. Indeed, if these treatments decreased severely the elongation of the exploratory root system (primary root and lateral roots) they had opposing effects on the root hair system. Although ACC increased the length and number of root hairs, the rate of N uptake and the transcript level of the N transporter BnNrt2.1 were markedly reduced. In contrast, the decrease in root hair length and number in AVG treated seedlings was overcompensated by an increase of N uptake and BnNrt2.1 gene expression. These root architectural changes demonstrated that BnNrt2.1 expression levels were more correlated to the changes of the exploratory root system than the changes of the root hair system. The difference between treatments in N transporters BnNrt1.1 and BnNrt2.1 gene expression is discussed with regard to presumed transport functions of BnNrt1.1 in relation to root elongation. PMID- 18287494 TI - The influences of associative cortices on cross-modal integration in the superior colliculus. PMID- 18287495 TI - Inhibitory gating of vibrissal inputs in the brainstem. AB - Trigeminal sensory nuclei are the first processing stage in the vibrissal system of rodents. They feature separate populations of thalamic projecting cells and a rich network of intersubnuclear connections, so that what is conveyed to the cortex by each of the ascending pathways of vibrissal information depends on local transactions that occur in the brainstem. In the present study, we examined the nature of these intersubnuclear connections by combining electrolytic lesions with electrophysiological recordings, retrograde labeling with in situ hybridization, and anterograde labeling with immunoelectron microscopy. Together, these different approaches provide conclusive evidence that the principal trigeminal nucleus receives inhibitory GABAergic projections from the caudal sector of the interpolaris subnucleus, and excitatory glutamatergic projections from the caudalis subnucleus. These results raise the possibility that, by controlling the activity of intersubnuclear projecting cells, brain regions that project to the spinal trigeminal nuclei may take an active part in selecting the type of vibrissal information that is conveyed through the lemniscal pathway. PMID- 18287496 TI - Calcium- and otoferlin-dependent exocytosis by immature outer hair cells. AB - Immature cochlear outer hair cells (OHCs) make transient synaptic contacts (ribbon synapses) with type I afferent nerve fibers, but direct evidence of synaptic vesicle exocytosis is still missing. We thus investigated calcium dependent exocytosis in murine OHCs at postnatal day 2 (P2)-P3, a developmental stage when calcium current maximum amplitude was the highest. By using time resolved patch-clamp capacitance measurements, we show that voltage step activation of L-type calcium channels triggers fast membrane capacitance increase. Capacitance increase displayed two kinetic components, which are likely to reflect two functionally distinct pools of synaptic vesicles, a readily releasable pool (RRP; tau = 79 ms) and a slowly releasable pool (tau = 870 ms). The RRP size and maximal release rate were estimated at approximately 1200 vesicles and approximately 15,000 vesicles/s, respectively. In addition, we found a linear relationship between capacitance increase and calcium influx, like in mature inner hair cells (IHCs). These results give strong support to the existence of efficient calcium-dependent neurotransmitter release in immature OHCs. Moreover, we show that immature OHCs, just like immature IHCs, are able to produce regenerative calcium-dependent action potentials that could trigger synaptic exocytosis in vivo. Finally, the evoked membrane capacitance increases were abolished in P2-P3 OHCs from mutant Otof-/- mice defective for otoferlin, despite normal calcium currents. We conclude that otoferlin, the putative major calcium sensor at IHC ribbon synapses, is essential to synaptic exocytosis in immature OHCs too. PMID- 18287497 TI - Noradrenergic modulation of electrical coupling in GABAergic networks of the hippocampus. AB - Noradrenergic modulation of cortical circuits is involved in information processing, regulation of higher functions, and prevention of epileptic activity. Here, we studied the effects of noradrenaline on the functional connectivity of GABAergic networks of the hippocampus and show that electrical synapses between interneurons are a novel target of noradrenergic modulation in vitro. Application of noradrenaline or of the selective beta-adrenergic agonist isoproterenol decreased gap junction-based coupling in paired recordings from stratum lacunosum moleculare interneurons by approximately 40%. Similar results were obtained after pharmacological stimulation of the adenylyl cyclase with forskolin. In contrast, the adenylyl cyclase antagonist MDL12330A [cis-N-(2 phenylcyclopentyl)azacyclotridec-1-en-2-amine] or the specific protein kinase A (PKA) inhibitor H89 (N-[2-(p-bromocinnamyl-amino)ethyl]-5-isoquinolinesulfonamide dihydrochloride) enhanced the basal strength of coupling by approximately 30%. In addition, PKA-mediated phosphorylation was critical for both isoproterenol- and forskolin-dependent regulation of coupling, because inclusion of the PKA antagonist KT5720 [(9S,10R,12R)-2,3,9,10,11,12-hexahydro-10-hydroxy-9-methyl-1 oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i][1,6]benzodiazocine 10-carboxylicacid hexyl ester] in the recording pipettes prevented modulation. Lastly, we studied the effects of beta-adrenergic modulation on mixed polysynaptic transmission within the GABAergic network. Isoproterenol depressed propagation of GABA(A) receptor-mediated synaptic currents, but did not change significantly direct GABAergic input, indicating that regulation of electrical coupling adds flexibility to the information flow generated by chemical synapses. In conclusion, activation of beta-adrenergic receptors in stratum lacunosum moleculare GABAergic networks reduces electrical synaptic transmission via a cAMP/PKA signaling cascade, and affects the degree of synaptic divergence within the circuit. We propose that this dynamic modulation and interplay between electrical and chemical synaptic transmission in GABAergic networks contributes to the tuning of memory processes in vivo, and prevents hypersynchronous activity. PMID- 18287498 TI - Prestimulus oscillatory activity in the alpha band predicts visual discrimination ability. AB - Although the resting and baseline states of the human electroencephalogram and magnetoencephalogram (MEG) are dominated by oscillations in the alpha band (approximately 10 Hz), the functional role of these oscillations remains unclear. In this study we used MEG to investigate how spontaneous oscillations in humans presented before visual stimuli modulate visual perception. Subjects had to report if there was a subtle difference in gray levels between two superimposed presented discs. We then compared the prestimulus brain activity for correctly (hits) versus incorrectly (misses) identified stimuli. We found that visual discrimination ability decreased with an increase in prestimulus alpha power. Given that reaction times did not vary systematically with prestimulus alpha power changes in vigilance are not likely to explain the change in discrimination ability. Source reconstruction using spatial filters allowed us to identify the brain areas accounting for this effect. The dominant sources modulating visual perception were localized around the parieto-occipital sulcus. We suggest that the parieto-occipital alpha power reflects functional inhibition imposed by higher level areas, which serves to modulate the gain of the visual stream. PMID- 18287499 TI - Complementary modulation of somatic inhibition by opioids and cannabinoids. AB - Somatic inhibition, which is critical for determining the spike output of principal cells, is mediated by two physiologically distinct classes of GABAergic interneurons called basket cells. In the hippocampus, despite both targeting the somatic membrane of CA1 pyramidal cells, these two classes of basket cells are active at different times. Differential modulation of these two types of basket cells could hence be important for regulating the activity patterns of CA1 pyramidal cells at very specific periods during ongoing activity. Indeed, cannabinoids selectively suppress the output of one class of basket cell. Whether opioids, another major modulator of inhibition in the hippocampus, also selectively suppress somatic inhibition is not known. Here, we show that basket cells are selectively modulated by either opioids or cannabinoids, but not both. We also find that basket cells are integrated into specific inhibitory subnetworks that are themselves under differential control of opioids and cannabinoids. Furthermore, because the two interneuron types are activated at different times, opioids and cannabinoids suppress different epochs of inhibition. This cell-type specific sensitivity to neuromodulators allows for a fine control of the temporal structure of hippocampal activity. PMID- 18287500 TI - An aggregate-inducing peripherin isoform generated through intron retention is upregulated in amyotrophic lateral sclerosis and associated with disease pathology. AB - The neuronal intermediate filament protein peripherin is a component of ubiquitinated inclusions and of axonal spheroids in amyotrophic lateral sclerosis (ALS). Overexpression of peripherin causes motor neuron degeneration in transgenic mice and variations within the peripherin gene have been identified in ALS cases. We have shown previously the abnormal expression of a neurotoxic peripherin splice variant in transgenic mice expressing mutant superoxide dismutase-1. These findings indicated that abnormalities of peripherin splicing may occur in ALS. In the current study, peripherin splice variants were identified by reverse transcription-PCR of human neuronal RNA and comparisons in expression made between control and ALS spinal cord using Western blot analysis and immunocytochemistry. Using this approach we have identified a novel peripherin transcript retaining introns 3 and 4 that results in a 28 kDa splice isoform, designated Per 28. Using an antibody specific to Per 28, we show that this isoform is expressed at low stoichiometric levels from the peripherin gene, however causes peripherin aggregation when its expression is upregulated. Importantly we show an upregulation of Per 28 expression in ALS compared with controls, at both the mRNA and protein levels, and that Per 28 is associated with disease pathology, specifically round inclusions. These findings are the first to establish that peripherin splicing abnormalities occur in ALS, generating aggregation-prone splice isoforms. PMID- 18287501 TI - Activation of the medial septum reverses age-related hippocampal encoding deficits: a place field analysis. AB - When a rat runs through a familiar environment, the hippocampus retrieves a previously stored spatial representation of the environment. When the environment is modified a new representation is seen, presumably corresponding to the hippocampus encoding the new information. The medial septum is hypothesized to modulate whether the hippocampus engages in retrieval or encoding. The cholinergic agonist carbachol was infused into the medial septum, and hippocampal CA1 place cells were recorded in freely moving rats. In a familiar environment, septal activation impaired the retrieval of a previously stored hippocampal place cell representation regardless of age. When the environment was changed, medial septal activation impaired the encoding process in young, but facilitated the encoding of the new information in aged rats. Moreover, the improved encoding was evident during a subsequent exposure to the modified environment 24 h later. The findings support the role the septum plays in modulating hippocampal retrieval/encoding states. Furthermore, our data indicate a mechanism of age related cognitive impairment. PMID- 18287502 TI - Ethanol consumption during early pregnancy alters the disposition of tangentially migrating GABAergic interneurons in the fetal cortex. AB - Consumption of alcohol (ethanol) during pregnancy can lead to developmental defects in the offspring, the most devastating being the constellation of symptoms collectively referred to as fetal alcohol syndrome (FAS). In the brain, a hallmark of FAS is abnormal cerebral cortical morphology consistent with insult during corticogenesis. Here, we report that exposure to a relatively low level of ethanol in utero (average maternal and fetal blood alcohol level of 25 mg/dl) promotes premature tangential migration into the cortical anlage of primordial GABAergic interneurons, including those originating in the medial ganglionic eminence (MGE). This ethanol-induced effect was evident in vivo at embryonic day 14.5 (E14.5) in GAD67 knock-in and BAC-Lhx6 embryos, as well as in vitro in isotypic telencephalic slice cocultures obtained from E14.5 embryos exposed to ethanol in utero. Analysis of heterotypic cocultures indicated that both cell intrinsic and -extrinsic factors contribute to the aberrant migratory profile of MGE-derived cells. In this light, we provide evidence for an interaction between ethanol exposure in utero and the embryonic GABAergic system. Exposure to ethanol in utero elevated the ambient level of GABA and increased the sensitivity to GABA of MGE-derived cells. Our results uncovered for the first time an effect of ethanol consumption during pregnancy on the embryonic development of GABAergic cortical interneurons. We propose that ethanol exerts its effect on the tangential migration of GABAergic interneurons extrinsically by modulating extracellular levels of GABA and intrinsically by altering GABA(A) receptor function. PMID- 18287503 TI - Ca2+/CaM controls Ca2+-dependent inactivation of NMDA receptors by dimerizing the NR1 C termini. AB - Ca2+ influx through NMDA receptors (NMDARs) leads to channel inactivation, which limits Ca2+ entry and protects against excitotoxicity. Extensive functional data suggests that this Ca2+-dependent inactivation (CDI) requires both calmodulin (CaM) binding to the C0 cassette of the NR1 subunit's C terminus (CT) and regulation by alpha-actinin-2, but a molecular understanding of CDI has been elusive. Here we used a number of methods to analyze the molecular nature of the interaction among CaM, alpha-actinin-2, and the NR1 CT. We found that a single CaM binds to two NR1 CTs in a Ca2+-dependent manner and promotes their reversible "dimerization." Expressed NMDARs containing NR1 concatamers in which the NR1 C termini are "uncoupled" display markedly reduced CDI. In contrast to current models, alpha-actinin-2 does not bind to the NR1 CT. We propose a new model for CDI in which the noncanonical Ca2+/CaM-dependent dimerization of the two NR1 subunits inactivates the channel by propagating a conformational change from the short NR1 CT to the nearby channel pore. PMID- 18287504 TI - Recurrent synaptic input and the timing of gamma-frequency-modulated firing of pyramidal cells during neocortical "UP" states. AB - Gamma (gamma) oscillation, a hallmark of cortical activity during sensory processing and cognition, occurs during persistent, self-sustained activity or "UP" states, which are thought to be maintained by recurrent synaptic inputs to pyramidal cells. During neocortical "UP" states, excitatory regular spiking (RS) (pyramidal) cells and inhibitory fast spiking (FS) (basket) cells fire with distinct phase distributions relative to the gamma oscillation in the local field potential. Evidence suggests that gamma-modulated RS --> FS input serves to synchronize the interneurons and hence to generate gamma-modulated FS --> RS drive. How RS --> RS recurrent input shapes both self-sustained activity and gamma-modulated phasic firing, although, is unclear. Here, we investigate this by reconstructing gamma-modulated synaptic input to RS cells using the conductance injection (dynamic clamp) technique in cortical slices. We find that, to show lifelike gamma-modulated firing, RS cells require strongly gamma-modulated, low latency inhibitory inputs from FS cells but little or no gamma-modulation from recurrent RS --> RS connections. We suggest that this demodulation of recurrent excitation, compared with inhibition, reflects several possible effects, including distributed propagation delays and integration of excitation over wider areas of cortex, and maximizes the capacity for representing information by the timing of recurrent excitation. PMID- 18287505 TI - Systemic vesicular stomatitis virus selectively destroys multifocal glioma and metastatic carcinoma in brain. AB - Metastatic tumors and malignant gliomas make up the majority of cancers in the brain. They are invariably fatal and there is currently no cure. From in vitro comparisons of a number of viruses, we selected one that appeared the best in selectively killing glioblastoma cells. This replication-competent virus, the glioma-adapted vesicular stomatis virus strain VSVrp30a, was used for in vivo tests with the underlying view that infection of tumor cells will lead to an increase in the number of viruses subsequently released to kill additional tumor cells. Intravenous injection of VSVrp30a expressing a green fluorescent protein reporter, rapidly targeted and destroyed multiple types of human and mouse tumors implanted in the mouse brain, including glioblastoma and mammary tumors. When tumors were implanted both in the brain and peripherally, emulating systemic cancer metastasis, tumors inside and outside the brain were simultaneously infected. Intranasal inoculation, leading to olfactory nerve transport of the virus into the brain, selectively infected and killed olfactory bulb tumors. Neither control cortical wounds nor transplanted normal mouse or human cells were targeted, indicating viral tumor selectivity. Control viruses, including pseudorabies, adeno-associated, or replication-deficient VSV, did not infect the brain tumor. Confocal laser time-lapse imaging through a cranial window showed that intravenous VSV infects the tumor at multiple sites and kills migrating tumor cells. Disrupted tumor vasculature, suggested by dye leakage, may be the port of entry for intravenously delivered VSV. Quantitative PCR analysis of how VSVrp30a selectively infected tumor cells suggested multiple mechanisms, including cell surface binding and internalization. PMID- 18287506 TI - Release of the styryl dyes from single synaptic vesicles in hippocampal neurons. AB - In small presynaptic boutons in brain, synaptic vesicles are thought not to merge with the plasma membrane when they release transmitter, but instead to close their fusion pores and survive intact for future use (kiss-and-run exocytosis). The strongest evidence for this idea is the slow and incomplete release of the fluorescent membrane marker, FM1-43 [N-(3-triethylammoniumpropyl)-4-(4 (dibutylamino)styryl) pyridinium dibromide], from single vesicles. We investigated the release of FM1-43 from sparse cultures of hippocampal neurons grown on coverslips with no glia. This allowed presynaptic boutons to be imaged at favorable signal-to-noise ratio. Sparingly stained boutons were imaged at high time resolution, while high-frequency electrical stimulation caused exocytosis. The release of FM1-43 was quantal and occurred in abrupt steps, each representing a single fusion event. The fluorescence of vesicle clusters traveling along axons had a distribution with the same quantal size, indicating that a vesicle releases all the dye it contains. In most fusion events, the time constant of dye release was <100 ms, and slower release was rarely observed. After exocytosis, no FM1-43 could be detected in the axon to either side of a bouton, indicating that dye was released before it could spread. Our results are consistent with synaptic vesicles fusing fully with the plasma membrane during high-frequency stimulation. PMID- 18287507 TI - Locomotor deficiencies and aberrant development of subtype-specific GABAergic interneurons caused by an unliganded thyroid hormone receptor alpha1. AB - Thyroid hormone (TH) deficiency during development causes severe and permanent neuronal damage, but the primary insult at the tissue level has remained unsolved. We have defined locomotor deficiencies in mice caused by a mutant thyroid hormone receptor alpha1 (TR alpha1) with potent aporeceptor activity attributable to reduced affinity to TH. This allowed identification of distinct functions that required either maternal supply of TH during early embryonic development or sufficient innate levels of hormone during late fetal development. In both instances, continued exposure to high levels of TH after birth and throughout life was needed. The hormonal dependencies correlated with severely delayed appearance of parvalbumin-immunoreactive GABAergic interneurons and increased numbers of calretinin-immunoreactive cells in the neocortex. This resulted in reduced numbers of fast spiking interneurons and defects in cortical network activity. The identification of locomotor deficiencies caused by insufficient supply of TH during fetal/perinatal development and their correlation with subtype-specific interneurons suggest a previously unknown basis for the neuronal consequences of endemic cretinism and untreated congenital hypothyroidism, and specifies TR alpha1 as the receptor isoform mediating these effects. PMID- 18287508 TI - An input-representing interneuron regulates spike timing and thereby phase switching in a motor network. AB - Despite the importance of spike-timing regulation in network functioning, little is known about this regulation at the cellular level. In the Aplysia feeding network, we show that interneuron B65 regulates the timing of the spike initiation of phase-switch neurons B64 and cerebral-buccal interneuron-5/6 (CBI 5/6), and thereby determines the identity of the neuron that acts as a protraction terminator. Previous work showed that B64 begins to fire before the end of protraction phase and terminates protraction in CBI-2-elicited ingestive, but not in CBI-2-elicited egestive programs, thus indicating that the spike timing and phase-switching function of B64 depend on the type of the central pattern generator (CPG)-elicited response rather than on the input used to activate the CPG. Here, we find that CBI-5/6 is a protraction terminator in egestive programs elicited by the esophageal nerve (EN), but not by CBI-2, thus indicating that, in contrast to B64, the spike timing and protraction-terminating function of CBI-5/6 depends on the input to the CPG rather than the response type. Interestingly, B65 activity also depends on the input in that B65 is highly active in EN-elicited programs, but not in CBI-2-elicited programs independent of whether the programs are ingestive or egestive. Notably, during EN-elicited egestive programs, hyperpolarization of B65 delays the onset of CBI-5/6 firing, whereas in CBI-2-elicited ingestive programs, B65 stimulation simultaneously advances CBI-5/6 firing and delays B64 firing, thereby substituting CBI-5/6 for B64 as the protraction terminator. Thus, we identified a neural mechanism that, in an input-dependent manner, regulates spike timing and thereby the functional role of specific neurons. PMID- 18287509 TI - Nonlinearities and contextual influences in auditory cortical responses modeled with multilinear spectrotemporal methods. AB - The relationship between a sound and its neural representation in the auditory cortex remains elusive. Simple measures such as the frequency response area or frequency tuning curve provide little insight into the function of the auditory cortex in complex sound environments. Spectrotemporal receptive field (STRF) models, despite their descriptive potential, perform poorly when used to predict auditory cortical responses, showing that nonlinear features of cortical response functions, which are not captured by STRFs, are functionally important. We introduce a new approach to the description of auditory cortical responses, using multilinear modeling methods. These descriptions simultaneously account for several nonlinearities in the stimulus-response functions of auditory cortical neurons, including adaptation, spectral interactions, and nonlinear sensitivity to sound level. The models reveal multiple inseparabilities in cortical processing of time lag, frequency, and sound level, and suggest functional mechanisms by which auditory cortical neurons are sensitive to stimulus context. By explicitly modeling these contextual influences, the models are able to predict auditory cortical responses more accurately than are STRF models. In addition, they can explain some forms of stimulus dependence in STRFs that were previously poorly understood. PMID- 18287510 TI - Microstructural correlates of infant functional development: example of the visual pathways. AB - The development of cognitive functions during childhood relies on several neuroanatomical maturation processes. Among these processes is myelination of the white matter pathways, which speeds up electrical conduction. Quantitative indices of such structural processes can be obtained in vivo with diffusion tensor imaging (DTI), but their physiological significance remains uncertain. Here, we investigated the microstructural correlates of early functional development by combining DTI and visual event-related potentials (VEPs) in 15 one to 4-month-old healthy infants. Interindividual variations of the apparent conduction speed, computed from the latency of the first positive VEP wave (P1), were significantly correlated with the infants' age and DTI indices measured in the optic radiations. This demonstrates that fractional anisotropy and transverse diffusivity are structural markers of functionally efficient myelination. Moreover, these indices computed along the optic radiations showed an early wave of maturation in the anterior region, with the posterior region catching up later in development, which suggests two asynchronous fronts of myelination in both the geniculocortical and corticogeniculate fibers. Thus, in addition to microstructural information, DTI provides noninvasive exquisite information on the functional development of the brain in human infants. PMID- 18287511 TI - Vesicle priming and recruitment by ubMunc13-2 are differentially regulated by calcium and calmodulin. AB - Ca2+ regulates multiple processes in nerve terminals, including synaptic vesicle recruitment, priming, and fusion. Munc13s, the mammalian homologs of Caenorhabditis elegans Unc13, are essential vesicle-priming proteins and contain multiple regulatory domains that bind second messengers such as diacylglycerol and Ca2+/calmodulin (Ca2+/CaM). Binding of Ca2+/CaM is necessary for the regulatory effect that allows Munc13-1 and ubMunc13-2 to promote short-term synaptic plasticity. However, the relative contributions of Ca2+ and Ca2+/CaM to vesicle priming and recruitment by Munc13 are not known. Here, we investigated the effect of Ca2+/CaM binding on ubMunc13-2 activity in chromaffin cells via membrane-capacitance measurements and a detailed simulation of the exocytotic machinery. Stimulating secretion under various basal Ca2+ concentrations from cells overexpressing either ubMunc13-2 or a ubMunc13-2 mutant deficient in CaM binding enabled a distinction between the effects of Ca2+ and Ca2+/CaM. We show that vesicle priming by ubMunc13-2 is Ca2+ dependent but independent of CaM binding to ubMunc13-2. However, Ca2+/CaM binding to ubMunc13-2 specifically promotes vesicle recruitment during ongoing stimulation. Based on the experimental data and our simulation, we propose that ubMunc13-2 is activated by two Ca2+-dependent processes: a slow activation mode operating at low Ca2+ concentrations, in which ubMunc13-2 acts as a priming switch, and a fast mode at high Ca2+ concentrations, in which ubMunc13-2 is activated in a Ca2+/CaM dependent manner and accelerates vesicle recruitment and maturation during stimulation. These different Ca2+ activation steps determine the kinetic properties of exocytosis and vesicle recruitment and can thus alter plasticity and efficacy of transmitter release. PMID- 18287512 TI - TGFbeta-Smad2 signaling regulates the Cdh1-APC/SnoN pathway of axonal morphogenesis. AB - Axon growth is critical to the establishment of neuronal connectivity. The E3 ubiquitin ligase Cdh1-anaphase-promoting complex (Cdh1-APC) and its substrate the transcriptional modulator SnoN form a cell-intrinsic pathway that orchestrates axonal morphogenesis in the mammalian brain. How the Cdh1-APC/SnoN pathway is controlled in the nervous system remained unknown. Here, we report that the TGFbeta-regulated signaling protein Smad2 plays a key role in regulating the Cdh1 APC/SnoN pathway in neurons. We find that Smad2 is expressed in primary granule neurons of the developing rat cerebellar cortex. The Smad signaling pathway is basally activated in neurons. Endogenous Smad2 is phosphorylated, localized in the nucleus, and forms a physical complex with endogenous SnoN in granule neurons. Inhibition of Smad signaling by several distinct approaches, including genetic knock-down of Smad2, stimulates axonal growth. Biochemical evidence and genetic epistasis analyses reveal that Smad2 acts upstream of SnoN in a shared pathway with Cdh1-APC in the control of axonal growth. Remarkably, Smad2 knock down also overrides the ability of adult rat myelin to inhibit axonal growth. Collectively, our findings define a novel function for Smad2 in regulation of the Cdh1-APC/SnoN cell-intrinsic pathway of axonal morphogenesis, and suggest that inhibition of Smad signaling may hold therapeutic potential in stimulating axonal growth after injury in the CNS. PMID- 18287513 TI - cAMP response element-binding protein 1 feedback loop is necessary for consolidation of long-term synaptic facilitation in Aplysia. AB - The transcription factor cAMP response element (CRE)-binding protein (CREB) plays an essential role in the induction of many forms of long-term synaptic plasticity. Levels of CREB1, the Aplysia homolog of CREB, show sustained elevations for several hours after the induction of long-term synaptic facilitation (LTF). Furthermore, CREB1 binds to the promoter of its own gene. These results suggest the existence of a CREB1-positive feedback loop that contributes to the consolidation of LTF. In the present study, we provide a detailed, quantitative characterization of the dynamics of CREB1 mRNA and protein as well as CREB1 phosphorylation after LTF induction. Injections of CRE oligonucleotides prevented the increase in CREB1 in response to 5-HT, corroborating the existence of the CREB1 feedback loop. This loop probably sustains CRE-dependent gene transcription, which remains elevated for at least 12 h after LTF induction. LTF is blocked by injection of CREB1 antibody after the induction phase, suggesting that the CREB1-positive feedback is required for consolidation of LTF. PMID- 18287514 TI - Activation of muscarinic receptors in rat bladder sensory pathways alters reflex bladder activity. AB - Antimuscarinic drugs affect bladder sensory symptoms such as urgency and frequency, presumably by acting on muscarinic acetylcholine receptors (mAChRs) located in bladder sensory pathways including primary afferent nerves and urothelium. However, the expression and the function of these receptors are not well understood. This study investigated the role of mAChRs in bladder sensory pathways in vivo in urethane anesthetized rats. Intravesical administration of the mAChR agonist oxotremorine methiodide (OxoM) elicited concentration-dependent excitatory and inhibitory effects on the frequency of voiding. These effects were blocked by intravesical administration of the mAChR antagonist atropine methyl nitrate (5 microM) and were absent in rats pretreated with capsaicin to desensitize C-fiber afferent nerves. Low concentrations of OxoM (5 microM) decreased voiding frequency by approximately 30%, an effect blunted by inhibiting nitric oxide (NO) synthesis with L-NAME (N(omega)-nitro-L-arginine methyl ester hydrochloride; 5 mg/kg; i.v.). High concentrations of OxoM (40 microM) increased voiding frequency by approximately 45%, an effect blunted by blocking purinergic receptors with PPADS (0.1-1 mM; intravesically). mAChR agonists stimulated release of ATP from cultured urothelial cells. Intravenous administration of OxoM (0.01-5 microg/kg) did not mimic the intravesical effects on voiding frequency. These results suggest that activation of mAChRs located near the luminal surface of the bladder affects voiding functions via mechanisms involving ATP and NO release presumably from the urothelium, that in turn could act on bladder C-fiber afferent nerves to alter their firing properties. These findings suggest that the urothelial-afferent nerve interactions can influence reflex voiding function. PMID- 18287515 TI - The good, the bad, and the cell type-specific roles of hypoxia inducible factor-1 alpha in neurons and astrocytes. AB - Hypoxia inducible factor-1alpha (HIF-1alpha) is a key regulator of oxygen homeostasis, because it is responsible for the regulation of genes involved in glycolysis, erythropoiesis, angiogenesis, and apoptosis. In the CNS, HIF-1alpha is stabilized by insults associated with hypoxia and ischemia. Because its many target genes mediate both adaptive and pathological processes, the role of HIF 1alpha in neuronal survival is debated. Although neuronal HIF-1alpha function has been the topic of several studies, the role of HIF-1alpha function in astrocytes has received much less attention. To characterize the role of HIF-1alpha in neurons and astrocytes, we induced loss of HIF-1alpha function specifically in neurons, astrocytes, or both cell types in neuron/astrocyte cocultures exposed to hypoxia. Although loss of HIF-1alpha function in neurons reduced neuronal viability during hypoxia, selective loss of HIF-1 function in astrocytes markedly protected neurons from hypoxic-induced neuronal death. Although the pathological processes induced by HIF-1alpha in astrocytes remain to be defined, induction of inducible nitric oxide synthase likely contributes to the pathological process. This study delineates, for the first time, a cell type-specific action for HIF 1alpha within astrocytes and neurons. PMID- 18287516 TI - Radiologic and nuclear events: contingency planning for hematologists/oncologists. AB - Untoward events involving radioactive material, either accidental or intentional, are potentially devastating. Hematologists and oncologists are uniquely suited to help manage radiation victims, as myelosuppression is a frequent complication of radiation exposure. In the aftermath of a large event, such as a nuclear detonation, there may be a national call for surge capacity that involves hematologists/oncologists across the country in the disaster response. In preparation, the National Marrow Donor Program and American Society for Blood and Marrow Transplantation have established the Radiation Injury Treatment Network (RITN), a voluntary consortium of transplant centers, donor centers, and umbilical cord blood banks. RITN is partnered with the Office of the Assistant Secretary for Preparedness and Response in the United States Department of Health and Human Services to develop treatment guidelines, educate healthcare professionals, coordinate situation response, and provide comprehensive evaluation and care for radiation injury victims. We outline the current plans for event response and describe scenarios, including catastrophic events that would require extensive support from hematologists/oncologists across the country. In addition, we highlight important reference resources and discuss current efforts to develop medical countermeasures against radiation toxicity. Practitioners and institutions across the country are encouraged to become involved and participate in the planning. PMID- 18287517 TI - A functional TNFRSF5 gene variant is associated with risk of lymphoma. AB - CD40 and its ligand, CD154, are major costimulatory molecules whose interactions are important in humoral and cellular immunity. We hypothesized that single nucleotide polymorphisms (SNPs) in TNFRSF5 and TNFSF5 encoding the CD40 and CD154 proteins, respectively, influence lymphoma risk, particularly a functional TNFRSF5 SNP (-1C>T, rs1883832) associated with reduced B-cell CD40 expression. TNFRSF5 and TNFSF5 SNPs were examined in a population-based case-control study of non-Hodgkin lymphoma (376 cases/801 controls with DNA), and compelling findings were followed up in 2 independent populations. Pooled analyses of all 3 case control studies (total N = 1776 non-Hodgkin lymphoma cases, N = 2482 controls) revealed an increased risk of follicular lymphoma (FL) associated with the TNFRSF5 -1TT genotype (odds ratio = 1.6; 95% confidence interval, 1.1-2.4). In addition, among women, an inverse association was found between the variant A allele for a TNFSF5 6809G>A SNP and FL risk (OR = .61; 95% CI, 0.36-0.98). In genotype-phenotype studies, significantly reduced circulating soluble CD40 was observed in TNFRSF5 -1TT compared with -1CC carriers. Further, dendritic cells from those with -1TT versus -1CC genotypes exhibited lower CD40 cell surface expression. These results suggest that the TNFRSF5 -1C>T polymorphism may increase FL susceptibility through mechanisms that hinder cellular immune responses. Further studies are needed to explore these findings. PMID- 18287518 TI - Protein networks supporting AP-3 function in targeting lysosomal membrane proteins. AB - The AP-3 adaptor complex targets selected transmembrane proteins to lysosomes and lysosome-related organelles. We reconstituted its preferred interaction with liposomes containing the ADP ribosylation factor (ARF)-1 guanosine triphosphatase (GTPase), specific cargo tails, and phosphatidylinositol-3 phosphate, and then we performed a proteomic screen to identify new proteins supporting its sorting function. We identified approximately 30 proteins belonging to three networks regulating either AP-3 coat assembly or septin polymerization or Rab7-dependent lysosomal transport. RNA interference shows that, among these proteins, the ARF-1 exchange factor brefeldin A-inhibited exchange factor 1, the ARF-1 GTPase activating protein 1, the Cdc42-interacting Cdc42 effector protein 4, an effector of septin-polymerizing GTPases, and the phosphatidylinositol-3 kinase IIIC3 are key components regulating the targeting of lysosomal membrane proteins to lysosomes in vivo. This analysis reveals that these proteins, together with AP-3, play an essential role in protein sorting at early endosomes, thereby regulating the integrity of these organelles. PMID- 18287519 TI - GEF-H1 couples nocodazole-induced microtubule disassembly to cell contractility via RhoA. AB - The RhoA GTPase plays a vital role in assembly of contractile actin-myosin filaments (stress fibers) and of associated focal adhesion complexes of adherent monolayer cells in culture. GEF-H1 is a microtubule-associated guanine nucleotide exchange factor that activates RhoA upon release from microtubules. The overexpression of GEF-H1 deficient in microtubule binding or treatment of HeLa cells with nocodazole to induce microtubule depolymerization results in Rho dependent actin stress fiber formation and contractile cell morphology. However, whether GEF-H1 is required and sufficient to mediate nocodazole-induced contractility remains unclear. We establish here that siRNA-mediated depletion of GEF-H1 in HeLa cells prevents nocodazole-induced cell contraction. Furthermore, the nocodazole-induced activation of RhoA and Rho-associated kinase (ROCK) that mediates phosphorylation of myosin regulatory light chain (MLC) is impaired in GEF-H1-depleted cells. Conversely, RhoA activation and contractility are rescued by reintroduction of siRNA-resistant GEF-H1. Our studies reveal a critical role for a GEF-H1/RhoA/ROCK/MLC signaling pathway in mediating nocodazole-induced cell contractility. PMID- 18287520 TI - Deletion of many yeast introns reveals a minority of genes that require splicing for function. AB - Splicing regulates gene expression and contributes to proteomic diversity in higher eukaryotes. However, in yeast only 283 of the 6000 genes contain introns and their impact on cell function is not clear. To assess the contribution of introns to cell function, we initiated large-scale intron deletions in yeast with the ultimate goal of creating an intron-free model eukaryote. We show that about one-third of yeast introns are not essential for growth. Only three intron deletions caused severe growth defects, but normal growth was restored in all cases by expressing the intronless mRNA from a heterologous promoter. Twenty percent of the intron deletions caused minor phenotypes under different growth conditions. Strikingly, the combined deletion of all introns from the 15 cytoskeleton-related genes did not affect growth or strain fitness. Together, our results show that although the presence of introns may optimize gene expression and provide benefit under stress, a majority of introns could be removed with minor consequences on growth under laboratory conditions, supporting the view that many introns could be phased out of Saccharomyces cerevisiae without blocking cell growth. PMID- 18287521 TI - DRhoGEF2 and diaphanous regulate contractile force during segmental groove morphogenesis in the Drosophila embryo. AB - Morphogenesis of the Drosophila embryo is associated with dynamic rearrangement of the actin cytoskeleton mediated by small GTPases of the Rho family. These GTPases act as molecular switches that are activated by guanine nucleotide exchange factors. One of these factors, DRhoGEF2, plays an important role in the constriction of actin filaments during pole cell formation, blastoderm cellularization, and invagination of the germ layers. Here, we show that DRhoGEF2 is equally important during morphogenesis of segmental grooves, which become apparent as tissue infoldings during mid-embryogenesis. Examination of DRhoGEF2 mutant embryos indicates a role for DRhoGEF2 in the control of cell shape changes during segmental groove morphogenesis. Overexpression of DRhoGEF2 in the ectoderm recruits myosin II to the cell cortex and induces cell contraction. At groove regression, DRhoGEF2 is enriched in cells posterior to the groove that undergo apical constriction, indicating that groove regression is an active process. We further show that the Formin Diaphanous is required for groove formation and strengthens cell junctions in the epidermis. Morphological analysis suggests that Dia regulates cell shape in a way distinct from DRhoGEF2. We propose that DRhoGEF2 acts through Rho1 to regulate acto-myosin constriction but not Diaphanous-mediated F-actin nucleation during segmental groove morphogenesis. PMID- 18287522 TI - Regulation of mitochondrial morphology by USP30, a deubiquitinating enzyme present in the mitochondrial outer membrane. AB - Recent studies have suggested that ubiquitination of mitochondrial proteins participates in regulating mitochondrial dynamics in mammalian cells, but it is unclear whether deubiquitination is involved in this process. Here, we identify human ubiquitin-specific protease 30 (USP30) as a deubiquitinating enzyme that is embedded in the mitochondrial outer membrane. Depletion of USP30 expression by RNA interference induced elongated and interconnected mitochondria, depending on the activities of the mitochondrial fusion factors mitofusins, without changing the expression levels of the key regulators for mitochondrial dynamics. Mitochondria were rescued from this abnormal phenotype by ectopic expression of USP30 in a manner dependent on its enzymatic activity. Our findings reveal that USP30 participates in the maintenance of mitochondrial morphology, a finding that provides new insight into the cellular function of deubiquitination. PMID- 18287523 TI - mDia2 induces the actin scaffold for the contractile ring and stabilizes its position during cytokinesis in NIH 3T3 cells. AB - mDia proteins are mammalian homologues of Drosophila diaphanous and belong to the formin family proteins that catalyze actin nucleation and polymerization. Although formin family proteins of nonmammalian species such as Drosophila diaphanous are essential in cytokinesis, whether and how mDia proteins function in cytokinesis remain unknown. Here we depleted each of the three mDia isoforms in NIH 3T3 cells by RNA interference and examined this issue. Depletion of mDia2 selectively increased the number of binucleate cells, which was corrected by coexpression of RNAi-resistant full-length mDia2. mDia2 accumulates in the cleavage furrow during anaphase to telophase, and concentrates in the midbody at the end of cytokinesis. Depletion of mDia2 induced contraction at aberrant sites of dividing cells, where contractile ring components such as RhoA, myosin, anillin, and phosphorylated ERM accumulated. Treatment with blebbistatin suppressed abnormal contraction, corrected localization of the above components, and revealed that the amount of F-actin at the equatorial region during anaphase/telophase was significantly decreased with mDia2 RNAi. These results demonstrate that mDia2 is essential in mammalian cell cytokinesis and that mDia2 induced F-actin forms a scaffold for the contractile ring and maintains its position in the middle of a dividing cell. PMID- 18287524 TI - p204 protein overcomes the inhibition of core binding factor alpha-1-mediated osteogenic differentiation by Id helix-loop-helix proteins. AB - Id proteins play important roles in osteogenic differentiation; however, the molecular mechanism remains unknown. In this study, we established that inhibitor of differentiation (Id) proteins, including Id1, Id2, and Id3, associate with core binding factor alpha-1 (Cbfa1) to cause diminished transcription of the alkaline phosphatase (ALP) and osteocalcin (OCL) gene, leading to less ALP activity and osteocalcin (OCL) production. Id acts by inhibiting the sequence specific binding of Cbfa1 to DNA and by decreasing the expression of Cbfa1 in cells undergoing osteogenic differentiation. p204, an interferon-inducible protein that interacts with both Cbfa1 and Id2, overcame the Id2-mediated inhibition of Cbfa1-induced ALP activity and OCL production. We show that 1) p204 disturbed the binding of Id2 to Cbfa1 and enabled Cbfa1 to bind to the promoters of its target genes and 2) that p204 promoted the translocation from nucleus to the cytoplasm and accelerated the degradation of Id2 by ubiquitin-proteasome pathway during osteogenesis. Nucleus export signal (NES) of p204 is required for the p204-enhanced cytoplasmic translocation and degradation of Id2, because a p204 mutant lacking NES lost these activities. Together, Cbfa1, p204, and Id proteins form a regulatory circuit and act in concert to regulate osteoblast differentiation. PMID- 18287525 TI - Localized RanGTP accumulation promotes microtubule nucleation at kinetochores in somatic mammalian cells. AB - Centrosomes are the major sites for microtubule nucleation in mammalian cells, although both chromatin- and kinetochore-mediated microtubule nucleation have been observed during spindle assembly. As yet, it is still unclear whether these pathways are coregulated, and the molecular requirements for microtubule nucleation at kinetochore are not fully understood. This work demonstrates that kinetochores are initial sites for microtubule nucleation during spindle reassembly after nocodazole. This process requires local RanGTP accumulation concomitant with delocalization from kinetochores of the hydrolysis factor RanGAP1. Kinetochore-driven microtubule nucleation is also activated after cold induced microtubule disassembly when centrosome nucleation is impaired, e.g., after Polo-like kinase 1 depletion, indicating that dominant centrosome activity normally masks the kinetochore-driven pathway. In cells with unperturbed centrosome nucleation, defective RanGAP1 recruitment at kinetochores after treatment with the Crm1 inhibitor leptomycin B activates kinetochore microtubule nucleation after cold. Finally, nascent microtubules associate with the RanGTP regulated microtubule-stabilizing protein HURP in both cold- and nocodazole treated cells. These data support a model for spindle assembly in which RanGTP dependent abundance of nucleation/stabilization factors at centrosomes and kinetochores orchestrates the contribution of the two spindle assembly pathways in mammalian cells. The complex of RanGTP, the export receptor Crm1, and nuclear export signal-bearing proteins regulates microtubule nucleation at kinetochores. PMID- 18287526 TI - Organization of the pre-autophagosomal structure responsible for autophagosome formation. AB - Autophagy induced by nutrient depletion is involved in survival during starvation conditions. In addition to starvation-induced autophagy, the yeast Saccharomyces cerevisiae also has a constitutive autophagy-like system, the Cvt pathway. Among 31 autophagy-related (Atg) proteins, the function of Atg17, Atg29, and Atg31 is required specifically for autophagy. In this study, we investigated the role of autophagy-specific (i.e., non-Cvt) proteins under autophagy-inducing conditions. For this purpose, we used atg11Delta cells in which the Cvt pathway is abrogated. The autophagy-unique proteins are required for the localization of Atg proteins to the pre-autophagosomal structure (PAS), the putative site for autophagosome formation, under starvation condition. It is likely that these Atg proteins function as a ternary complex, because Atg29 and Atg31 bind to Atg17. The Atg1 kinase complex (Atg1-Atg13) is also essential for recruitment of Atg proteins to the PAS. The assembly of Atg proteins to the PAS is observed only under autophagy inducing conditions, indicating that this structure is specifically involved in autophagosome formation. Our results suggest that Atg1 complex and the autophagy unique Atg proteins cooperatively organize the PAS in response to starvation signals. PMID- 18287527 TI - Cell cycle-dependent binding of HMGN proteins to chromatin. AB - Throughout the cell cycle, the histones remain associated with DNA, but the repertoire of proteins associated with the chromatin fiber continuously changes. The chromatin interaction of HMGNs, a family of nucleosome binding proteins that modulates the structure and activity of chromatin, during the cell cycle is controversial. Immunofluorescence studies demonstrated that HMGNs are not associated with chromatin, whereas live cell imaging indicated that they are present in mitotic chromosomes. To resolve this controversy, we examined the organization of wild-type and mutated HMGN1 and HMGN2 proteins in the cell nucleus by using immunofluorescence studies, live cell imaging, gel mobility shift assays, and bimolecular fluorescence complementation (BiFC). We find that during interphase, HMGNs bind specifically to nucleosomes and form homodimeric complexes that yield distinct BiFC signals. In metaphase, the nucleosomal binding domain of the protein is inactivated, and the proteins associate with chromatin with low affinity as monomers, and they do not form specific complexes. Our studies demonstrate that the mode of binding of HMGNs to chromatin is cell cycle dependent. PMID- 18287528 TI - The cargo receptors Surf4, endoplasmic reticulum-Golgi intermediate compartment (ERGIC)-53, and p25 are required to maintain the architecture of ERGIC and Golgi. AB - Rapidly cycling proteins of the early secretory pathway can operate as cargo receptors. Known cargo receptors are abundant proteins, but it remains mysterious why their inactivation leads to rather limited secretion phenotypes. Studies of Surf4, the human orthologue of the yeast cargo receptor Erv29p, now reveal a novel function of cargo receptors. Surf4 was found to interact with endoplasmic reticulum-Golgi intermediate compartment (ERGIC)-53 and p24 proteins. Silencing Surf4 together with ERGIC-53 or silencing the p24 family member p25 induced an identical phenotype characterized by a reduced number of ERGIC clusters and fragmentation of the Golgi apparatus without effect on anterograde transport. Live imaging showed decreased stability of ERGIC clusters after knockdown of p25. Silencing of Surf4/ERGIC-53 or p25 resulted in partial redistribution of coat protein (COP) I but not Golgi matrix proteins to the cytosol and partial resistance of the cis-Golgi to brefeldin A. These findings imply that cargo receptors are essential for maintaining the architecture of ERGIC and Golgi by controlling COP I recruitment. PMID- 18287529 TI - The mood stabilizer valproate inhibits both inositol- and diacylglycerol signaling pathways in Caenorhabditis elegans. AB - The antiepileptic valproate (VPA) is widely used in the treatment of bipolar disorder, although the mechanism of its action in the disorder is unclear. We show here that VPA inhibits both inositol phosphate and diacylglycerol (DAG) signaling in Caenorhabditis elegans. VPA disrupts two behaviors regulated by the inositol-1,4,5-trisphosphate (IP(3)): defecation and ovulation. VPA also inhibits two activities regulated by DAG signaling: acetylcholine release and egg laying. The effects of VPA on DAG signaling are relieved by phorbol ester, a DAG analogue, suggesting that VPA acts to inhibit DAG production. VPA reduces levels of DAG and inositol-1-phosphate, but phosphatidylinositol-4,5-bisphosphate (PIP(2)) is slightly increased, suggesting that phospholipase C-mediated hydrolysis of PIP(2) to form DAG and IP(3) is defective in the presence of VPA. PMID- 18287530 TI - Tight junction proteins claudin-2 and -12 are critical for vitamin D-dependent Ca2+ absorption between enterocytes. AB - Ca(2+) is absorbed across intestinal epithelial monolayers via transcellular and paracellular pathways, and an active form of vitamin D(3), 1alpha,25 dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], is known to promote intestinal Ca(2+) absorption. However, the molecules driving the paracellular Ca(2+) absorption and its vitamin D dependency remain obscure. Because the tight junction proteins claudins are suggested to form paracellular channels for selective ions between neighboring cells, we hypothesized that specific intestinal claudins might facilitate paracellular Ca(2+) transport and that expression of these claudins could be induced by 1alpha,25(OH)(2)D(3). Herein, we show, by using RNA interference and overexpression strategies, that claudin-2 and claudin-12 contribute to Ca(2+) absorption in intestinal epithelial cells. We also provide evidence showing that expression of claudins-2 and -12 is up regulated in enterocytes in vitro and in vivo by 1alpha,25(OH)(2)D(3) through the vitamin D receptor. These findings strongly suggest that claudin-2- and/or claudin-12-based tight junctions form paracellular Ca(2+) channels in intestinal epithelia, and they highlight a novel mechanism behind vitamin D-dependent calcium homeostasis. PMID- 18287531 TI - Rab8 regulates basolateral secretory, but not recycling, traffic at the recycling endosome. AB - Rab8 is a monomeric GTPase that regulates the delivery of newly synthesized proteins to the basolateral surface in polarized epithelial cells. Recent publications have demonstrated that basolateral proteins interacting with the mu1 B clathrin adapter subunit pass through the recycling endosome (RE) en route from the TGN to the plasma membrane. Because Rab8 interacts with these basolateral proteins, these findings raise the question of whether Rab8 acts before, at, or after the RE. We find that Rab8 overexpression during the formation of polarity in MDCK cells, disrupts polarization of the cell, explaining how Rab8 mutants can disrupt basolateral endocytic and secretory traffic. However, once cells are polarized, Rab8 mutants cause mis-sorting of newly synthesized basolateral proteins such as VSV-G to the apical surface, but do not cause mis-sorting of membrane proteins already at the cell surface or in the endocytic recycling pathway. Enzymatic ablation of the RE also prevents traffic from the TGN from reaching the RE and similarly results in mis-sorting of newly synthesized VSV-G. We conclude that Rab8 regulates biosynthetic traffic through REs to the plasma membrane, but not trafficking of endocytic cargo through the RE. The data are consistent with a model in which Rab8 functions in regulating the delivery of TGN derived cargo to REs. PMID- 18287532 TI - Cholesterol loss enhances TrkB signaling in hippocampal neurons aging in vitro. AB - Binding of the neurotrophin brain-derived neurotrophic factor (BDNF) to the TrkB receptor is a major survival mechanism during embryonic development. In the aged brain, however, BDNF levels are low, suggesting that if TrkB is to play a role in survival at this stage additional mechanisms must have developed. We here show that TrkB activity is most robust in the hippocampus of 21-d-old BDNF-knockout mice as well as in old, wild-type, and BDNF heterozygous animals. Moreover, robust TrkB activity is evident in old but not young hippocampal neurons differentiating in vitro in the absence of any exogenous neurotrophin and also in neurons from BDNF -/- embryos. Age-associated increase in TrkB activity correlated with a mild yet progressive loss of cholesterol. This, in turn, correlated with increased expression of the cholesterol catabolic enzyme cholesterol 24-hydroxylase. Direct cause-effect, cholesterol loss-high TrkB activity was demonstrated by pharmacological means and by manipulating the levels of cholesterol 24-hydroxylase. Because reduced levels of cholesterol and increased expression of choleseterol-24-hydroxylase were also observed in the hippocampus of aged mice, changes in cellular cholesterol content may be used to modulate receptor activity strength in vivo, autonomously or as a way to complement the natural decay of neurotrophin production. PMID- 18287533 TI - Arp2 links autophagic machinery with the actin cytoskeleton. AB - Macroautophagy involves lysosomal/vacuolar elimination of long-lived proteins and entire organelles from the cytosol. The process begins with formation of a double membrane vesicle that sequesters bulk cytoplasm, or a specific cargo destined for lysosomal/vacuolar delivery. The completed vesicle fuses with the lysosome/vacuole limiting membrane, releasing its content into the organelle lumen for subsequent degradation and recycling of the resulting macromolecules. A majority of the autophagy-related (Atg) proteins are required at the step of vesicle formation. The integral membrane protein Atg9 cycles between certain intracellular compartments and the vesicle nucleation site, presumably to supply membranes necessary for macroautophagic vesicle formation. In this study we have tracked the movement of Atg9 over time in living cells by using real-time fluorescence microscopy. Our results reveal that an actin-related protein, Arp2, briefly colocalizes with Atg9 and directly regulates the dynamics of Atg9 movement. We propose that proteins of the Arp2/3 complex regulate Atg9 transport for specific types of autophagy. PMID- 18287534 TI - FBXO25-associated nuclear domains: a novel subnuclear structure. AB - Skp1, Cul1, Rbx1, and the FBXO25 protein form a functional ubiquitin ligase complex. Here, we investigate the cellular distribution of FBXO25 and its colocalization with some nuclear proteins by using immunochemical and biochemical approaches. FBXO25 was monitored with affinity-purified antibodies raised against the recombinant fragment spanning residues 2-62 of the FBXO25 sequence. FBXO25 protein was expressed in all mouse tissues tested except striated muscle, as indicated by immunoblot analysis. Confocal analysis revealed that the endogenous FBXO25 was partially concentrated in a novel dot-like nuclear domain that is distinct from clastosomes and other well-characterized structures. These nuclear compartments contain a high concentration of ubiquitin conjugates and at least two other components of the ubiquitin-proteasome system: 20S proteasome and Skp1. We propose to name these compartments FBXO25-associated nuclear domains. Interestingly, inhibition of transcription by actinomycin D or heat-shock treatment drastically affected the nuclear organization of FBXO25-containing structures, indicating that they are dynamic compartments influenced by the transcriptional activity of the cell. Also, we present evidences that an FBXO25 dependent ubiquitin ligase activity prevents aggregation of recombinant polyglutamine-containing huntingtin protein in the nucleus of human embryonic kidney 293 cells, suggesting that this protein can be a target for the nuclear FBXO25 mediated ubiquitination. PMID- 18287535 TI - Neuronal death by oxidative stress involves activation of FOXO3 through a two-arm pathway that activates stress kinases and attenuates insulin-like growth factor I signaling. AB - Oxidative stress kills neurons by stimulating FOXO3, a transcription factor whose activity is inhibited by insulin-like growth factor I (IGF-I), a wide-spectrum neurotrophic signal. Because recent evidence has shown that oxidative stress blocks neuroprotection by IGF-I, we examined whether attenuation of IGF-I signaling is linked to neuronal death by oxidative stress, as both events may contribute to neurodegeneration. We observed that in neurons, activation of FOXO3 by a burst of oxidative stress elicited by 50 muM hydrogen peroxide (H(2)O(2)) recruited a two-pronged pathway. A first, rapid arm attenuated AKT inhibition of FOXO3 through p38 MAPK-mediated blockade of IGF-I stimulation of AKT. A second delayed arm involved activation of FOXO3 by Jun-kinase 2 (JNK2). Notably, blockade of IGF-I signaling through p38 MAPK was necessary for JNK2 to activate FOXO3, unveiling a competitive regulatory interplay between the two arms onto FOXO3 activity. Therefore, an abrupt rise in oxidative stress activates p38 MAPK to tilt the balance in a competitive AKT/JNK2 regulation of FOXO3 toward its activation, eventually leading to neuronal death. In view of previous observations linking attenuation of IGF-I signaling to other causes of neuronal death, these findings suggest that blockade of trophic input is a common step in neuronal death. PMID- 18287536 TI - The Rim101 pathway is involved in Rsb1 expression induced by altered lipid asymmetry. AB - Biological membranes consist of lipid bilayers. The lipid compositions between the two leaflets of the plasma membrane differ, generating lipid asymmetry. Maintenance of proper lipid asymmetry is physiologically quite important, and its collapse induces several cellular responses including apoptosis and platelet coagulation. Thus, a change in lipid asymmetry must be restored to maintain "lipid asymmetry homeostasis." However, to date no lipid asymmetry-sensing proteins or any related downstream signaling pathways have been identified. We recently demonstrated that expression of the putative yeast sphingoid long-chain base transporter/translocase Rsb1 is induced when glycerophospholipid asymmetry is altered. Using mutant screening, we determined that the pH-responsive Rim101 pathway, the protein kinase Mck1, and the transcription factor Mot3 all act in lipid asymmetry signaling, and that the Rim101 pathway was activated in response to a change in lipid asymmetry. The activated transcription factor Rim101 induces Rsb1 expression via repression of another transcription repressor, Nrg1. Changes in lipid asymmetry are accompanied by cell surface exposure of negatively charged phospholipids; we speculate that the Rim101 pathway recognizes the surface charges. PMID- 18287537 TI - Paralemmin-1, a modulator of filopodia induction is required for spine maturation. AB - Dendritic filopodia are thought to participate in neuronal contact formation and development of dendritic spines; however, molecules that regulate filopodia extension and their maturation to spines remain largely unknown. Here we identify paralemmin-1 as a regulator of filopodia induction and spine maturation. Paralemmin-1 localizes to dendritic membranes, and its ability to induce filopodia and recruit synaptic elements to contact sites requires protein acylation. Effects of paralemmin-1 on synapse maturation are modulated by alternative splicing that regulates spine formation and recruitment of AMPA-type glutamate receptors. Paralemmin-1 enrichment at the plasma membrane is subject to rapid changes in neuronal excitability, and this process controls neuronal activity-driven effects on protrusion expansion. Knockdown of paralemmin-1 in developing neurons reduces the number of filopodia and spines formed and diminishes the effects of Shank1b on the transformation of existing filopodia into spines. Our study identifies a key role for paralemmin-1 in spine maturation through modulation of filopodia induction. PMID- 18287538 TI - Bap31 is an itinerant protein that moves between the peripheral endoplasmic reticulum (ER) and a juxtanuclear compartment related to ER-associated Degradation. AB - Certain endoplasmic reticulum (ER)-associated degradation (ERAD) substrates with transmembrane domains are segregated from other ER proteins and sorted into a juxtanuclear subcompartment, known as the ER quality control compartment. Bap31 is an ER protein with three transmembrane domains, and it is assumed to be a cargo receptor for ER export of some transmembrane proteins, especially those prone to ERAD. Here, we show that Bap31 is a component of the ER quality control compartment and that it moves between the peripheral ER and a juxtanuclear ER or ER-related compartment distinct from the conventional ER-Golgi intermediate compartment. The third and second transmembrane domains of Bap31 are principally responsible for the movement to and recycling from the juxtanuclear region, respectively. This cycling was blocked by depolymerization of microtubules and disruption of dynein-dynactin function. Overexpression of Sar1p and Arf1 mutants affected Bap31 cycling, suggesting that this cycling pathway is related to the conventional vesicular transport pathways. PMID- 18287540 TI - GRIM-19 is essential for maintenance of mitochondrial membrane potential. AB - GRIM-19 was found to copurify with complex I of mitochondrial respiratory chain and subsequently was demonstrated to be involved in complex I assembly and activity. To further understand its function in complex I, we dissected its functional domains by generating a number of deletion, truncation, and point mutants. The mitochondrial localization sequences were located at the N-terminus. Strikingly, deletion of residues 70-80, 90-100, or the whole C-terminal region (70-144) led to a loss of mitochondrial transmembrane potential (DeltaPsim). However, similar deletions of another two complex I subunits, NDUFA9 and NDUFS3, did not show such effect. We also found that deletion of the last 10 residues affected GRIM-19's ability to be assembled to complex I. We constructed a dominant-negative mutant containing the N-terminal 60 and the last C-terminal 10 residues, which could be assembled into complex I, but failed to maintain normal DeltaPsim. Cells overexpressing this mutant did not spontaneously undergo cell death, but were sensitized to apoptosis induced by cell death agents. Our results demonstrate that GRIM-19 is required for electron transfer activity of complex I, and disruption of DeltaPsim by GRIM-19 mutants enhances the cells' sensitivity to apoptotic stimuli. PMID- 18287539 TI - Yeast ARV1 is required for efficient delivery of an early GPI intermediate to the first mannosyltransferase during GPI assembly and controls lipid flow from the endoplasmic reticulum. AB - Glycosylphosphatidylinositol (GPI), covalently attached to many eukaryotic proteins, not only acts as a membrane anchor but is also thought to be a sorting signal for GPI-anchored proteins that are associated with sphingolipid and sterol enriched domains. GPI anchors contain a core structure conserved among all species. The core structure is synthesized in two topologically distinct stages on the leaflets of the endoplasmic reticulum (ER). Early GPI intermediates are assembled on the cytoplasmic side of the ER and then are flipped into the ER lumen where a complete GPI precursor is synthesized and transferred to protein. The flipping process is predicted to be mediated by a protein referred as flippase; however, its existence has not been proven. Here we show that yeast Arv1p is an important protein required for the delivery of an early GPI intermediate, GlcN-acylPI, to the first mannosyltransferase of GPI synthesis in the ER lumen. We also provide evidence that ARV1 deletion and mutations in other proteins involved in GPI anchor synthesis affect inositol phosphorylceramide synthesis as well as the intracellular distribution and amounts of sterols, suggesting a role of GPI anchor synthesis in lipid flow from the ER. PMID- 18287541 TI - Ste20-related protein kinase LOSK (SLK) controls microtubule radial array in interphase. AB - Interphase microtubules are organized into a radial array with centrosome in the center. This organization is a subject of cellular regulation that can be driven by protein phosphorylation. Only few protein kinases that regulate microtubule array in interphase cells have been described. Ste20-like protein kinase LOSK (SLK) was identified as a microtubule and centrosome-associated protein. In this study we have shown that the inhibition of LOSK activity by dominant-negative mutant K63R-DeltaT or by LOSK depletion with RNAi leads to unfocused microtubule arrangement. Microtubule disorganization is prominent in Vero, CV-1, and CHO-K1 cells but less distinct in HeLa cells. The effect is a result neither of microtubule stabilization nor of centrosome disruption. In cells with suppressed LOSK activity centrosomes are unable to anchor or to cap microtubules, though they keep nucleating microtubules. These centrosomes are depleted of dynactin. Vero cells overexpressing K63R-DeltaT have normal dynactin "comets" at microtubule ends and unaltered morphology of Golgi complex but are unable to polarize it at the wound edge. We conclude that protein kinase LOSK is required for radial microtubule organization and for the proper localization of Golgi complex in various cell types. PMID- 18287542 TI - The clathrin adaptor Gga2p is a phosphatidylinositol 4-phosphate effector at the Golgi exit. AB - Phosphatidylinositol 4-phosphate (PI(4)P) is a key regulator of membrane transport required for the formation of transport carriers from the trans-Golgi network (TGN). The molecular mechanisms of PI(4)P signaling in this process are still poorly understood. In a search for PI(4)P effector molecules, we performed a screen for synthetic lethals in a background of reduced PI(4)P and found the gene GGA2. Our analysis uncovered a PI(4)P-dependent recruitment of the clathrin adaptor Gga2p to the TGN during Golgi-to-endosome trafficking. Gga2p recruitment to liposomes is stimulated both by PI(4)P and the small GTPase Arf1p in its active conformation, implicating these two molecules in the recruitment of Gga2p to the TGN, which ultimately controls the formation of clathrin-coated vesicles. PI(4)P binding occurs through a phosphoinositide-binding signature within the N terminal VHS domain of Gga2p resembling a motif found in other clathrin interacting proteins. These data provide an explanation for the TGN-specific membrane recruitment of Gga2p. PMID- 18287543 TI - Similar properties of transient, persistent, and resurgent Na currents in GABAergic and non-GABAergic vestibular nucleus neurons. AB - Sodium currents in fast firing neurons are tuned to support sustained firing rates >50-60 Hz. This is typically accomplished with fast channel kinetics and the ability to minimize the accumulation of Na channels into inactivated states. Neurons in the medial vestibular nuclei (MVN) can fire at exceptionally high rates, but their Na currents have never been characterized. In this study, Na current kinetics and voltage-dependent properties were compared in two classes of MVN neurons with distinct firing properties. Non-GABAergic neurons (fluorescently labeled in YFP-16 transgenic mice) have action potentials with faster rise and fall kinetics and sustain higher firing rates than GABAergic neurons (fluorescently labeled in GIN transgenic mice). A previous study showed that these neurons express a differential balance of K currents. To determine whether the Na currents in these two populations were different, their kinetics and voltage-dependent properties were measured in acutely dissociated neurons from 24 to 40-day-old mice. All neurons expressed persistent Na currents and large transient Na currents with resurgent kinetics tuned for fast firing. No differences were found between the Na currents expressed in GABAergic and non GABAergic MVN neurons, suggesting that differences in properties of these neurons are tuned by their K currents. PMID- 18287545 TI - What's that sound? Auditory area CLM encodes stimulus surprise, not intensity or intensity changes. AB - High-level sensory neurons encoding natural stimuli are not well described by linear models operating on the time-varying stimulus intensity. Here we show that firing rates of neurons in a secondary sensory forebrain area can be better modeled by linear functions of how surprising the stimulus is. We modeled auditory neurons in the caudal lateral mesopallium (CLM) of adult male zebra finches under urethane anesthesia with linear filters convolved not with stimulus intensity, but with stimulus surprise. Surprise was quantified as the logarithm of the probability of the stimulus given the local recent stimulus history and expectations based on conspecific song. Using our surprise method, the predictions of neural responses to conspecific song improved by 67% relative to those obtained using stimulus intensity. Similar prediction improvements cannot be replicated by assuming CLM performs derivative detection. The explanatory power of surprise increased from the midbrain through the primary forebrain and to CLM. When the stimulus presented was a random synthetic ripple noise, CLM neurons (but not neurons in lower auditory areas) were best described as if they were expecting conspecific song, finding the inconsistencies between birdsong and noise surprising. In summary, spikes in CLM neurons indicate stimulus surprise more than they indicate stimulus intensity features. The concept of stimulus surprise may be useful for modeling neural responses in other higher-order sensory areas whose functions have been poorly understood. PMID- 18287544 TI - Spatial heterogeneity of cortical receptive fields and its impact on multisensory interactions. AB - Investigations of multisensory processing at the level of the single neuron have illustrated the importance of the spatial and temporal relationship of the paired stimuli and their relative effectiveness in determining the product of the resultant interaction. Although these principles provide a good first-order description of the interactive process, they were derived by treating space, time, and effectiveness as independent factors. In the anterior ectosylvian sulcus (AES) of the cat, previous work hinted that the spatial receptive field (SRF) architecture of multisensory neurons might play an important role in multisensory processing due to differences in the vigor of responses to identical stimuli placed at different locations within the SRF. In this study the impact of SRF architecture on cortical multisensory processing was investigated using semichronic single-unit electrophysiological experiments targeting a multisensory domain of the cat AES. The visual and auditory SRFs of AES multisensory neurons exhibited striking response heterogeneity, with SRF architecture appearing to play a major role in the multisensory interactions. The deterministic role of SRF architecture was tightly coupled to the manner in which stimulus location modulated the responsiveness of the neuron. Thus multisensory stimulus combinations at weakly effective locations within the SRF resulted in large (often superadditive) response enhancements, whereas combinations at more effective spatial locations resulted in smaller (additive/subadditive) interactions. These results provide important insights into the spatial organization and processing capabilities of cortical multisensory neurons, features that may provide important clues as to the functional roles played by this area in spatially directed perceptual processes. PMID- 18287546 TI - Proprioceptive and cutaneous representations in the rat ventral posterolateral thalamus. AB - Determining how and where proprioceptive information is represented in the rat ventral posterolateral (VPL) is important in allowing us to further investigate how this sense is utilized during motor control and learning. Here we demonstrate using electrophysiological techniques that the rostral portion of the rat VPL nucleus (rVPL, -2 to -2.5 mm bregma) carries a large amount of proprioceptive information. Caudal to this region is a zone where the cutaneous receptive fields are focal (mVPL for middle VPL, -2.5 to -3.2 mm bregma) with a fine topographic map of the fore- and hindlimbs. The forepaw is represented with digit 1 medial and each subsequent digit increasingly lateral, all of which are dorsal to the pads. The caudal VPL (cVPL, -3.2 to -4.0 mm bregma) has broad receptive fields and is the target of lamina 1 and lamina 2, as well as the dorsal column nuclei, and may represent the flow of nociceptive information through the VPL. Thus we propose that the VPL may be thought of as three subnuclei-the rostral, middle, and caudal VPL-each carrying preferentially a different modality of information. This pattern of information flow through the rat VPL is similar, although apparently rotated, to that of many primates, indicating that these regions in the rat (rVPL, mVPL, and cVPL) have become further differentiated in primates where they are seen as separate nuclei (VPS, VPL, and VPI/VMpo). PMID- 18287547 TI - Differential modulation of neural network and pacemaker activity underlying eupnea and sigh-breathing activities. AB - Many networks generate distinct rhythms with multiple frequency and amplitude characteristics. The respiratory network in the pre-Botzinger complex (pre-Bot) generates both the low-frequency, large-amplitude sigh rhythm and a faster, smaller-amplitude eupneic rhythm. Could the same set of pacemakers generate both rhythms? Here we used an in vitro respiratory brainslice preparation. We describe a subset of synaptically isolated pacemakers that spontaneously generate two distinct bursting patterns. These two patterns resemble network activity including sigh-like bursts that occur at low frequencies and have large amplitudes and eupneic-like bursts with higher frequency and smaller amplitudes. Cholinergic neuromodulation altered the network and pacemaker bursting: fictive sigh frequency is increased dramatically, whereas fictive eupneic frequency is drastically lowered. The data suggest that timing and amplitude characteristics of fictive eupneic and sigh rhythms are set by the same set of pacemakers that are tuned by changes in the neuromodulatory state. PMID- 18287548 TI - Recording temperature affects the excitability of mouse superficial dorsal horn neurons, in vitro. AB - Superficial dorsal horn (SDH) neurons in laminae I-II of the spinal cord play an important role in processing noxious stimuli. These neurons represent a heterogeneous population and are divided into various categories according to their action potential (AP) discharge during depolarizing current injection. We recently developed an in vivo mouse preparation to examine functional aspects of nociceptive processing and AP discharge in SDH neurons and to extend investigation of pain mechanisms to the genetic level of analysis. Not surprisingly, some in vivo data obtained at body temperature (37 degrees C) differed from those generated at room temperature (22 degrees C) in spinal cord slices. In the current study we examine how temperature influences SDH neuron properties by making recordings at 22 and 32 degrees C in transverse spinal cord slices prepared from L3-L5 segments of adult mice (C57Bl/6). Patch-clamp recordings (KCH(3)SO(4) internal) were made from visualized SDH neurons. At elevated temperature all SDH neurons had reduced input resistance and smaller, briefer APs. Resting membrane potential and AP afterhyperpolarization amplitude were temperature sensitive only in subsets of the SDH population. Notably, elevated temperature increased the prevalence of neurons that did not discharge APs during current injection. These reluctant firing neurons expressed a rapid A type potassium current, which is enhanced at higher temperatures and thus restrains AP discharge. When compared with previously published whole cell recordings obtained in vivo (37 degrees C) our results suggest that, on balance, in vitro data collected at elevated temperature more closely resemble data collected under in vivo conditions. PMID- 18287549 TI - Effects of visual and auditory feedback on sensorimotor circuits in the basal ganglia. AB - Previous work using visual feedback has identified two distinct sensorimotor circuits in the basal ganglia (BG): one that scaled with the duration of force and one that scaled with the rate of change of force. The present study compared functional MRI signal changes in the BG during a grip force task using either visual or auditory feedback to determine whether the BG nuclei process auditory and visual feedback similarly. We confirmed the same two sensorimotor circuits in the BG. Activation in the striatum and external globus pallidus (GPe) scaled linearly with the duration of force under visual and auditory feedback conditions, with similar slopes and intercepts across feedback type. The pattern of signal change for the internal globus pallidus (GPi) and subthalamic nucleus (STN) was nonlinear and parameters of the exponential function were altered by feedback type. Specifically, GPi and STN activation decreased exponentially with the rate of change of force. The rate constant and asymptote of the exponential functions for GPi and STN were greater during auditory than visual feedback. In a comparison of the BOLD signal between BG regions, GPe had the highest percentage of variance accounted for and this effect was preserved for both feedback types. These new findings suggest that neuronal activity of specific BG nuclei is affected by whether the feedback is derived from visual or auditory inputs. Also, the data are consistent with the hypothesis that the GPe has a high level of information convergence from other BG nuclei, which is preserved across different sensory feedback modalities. PMID- 18287550 TI - Interactions with compliant loads alter stretch reflex gains but not intermuscular coordination. AB - The human motor system regulates arm mechanics to produce stable postures during interactions with different physical environments. This occurs partly via involuntary mechanisms, including stretch reflexes. Previous single-joint studies demonstrated enhanced reflex sensitivity during interactions with compliant environments, suggesting reflex gain increases to enhance limb stability when that stability is not provided by the environment. This study examined whether similar changes in reflex gain are present throughout the limb following perturbations that simultaneously influence multiple joints. Furthermore, we investigated whether any observed modulation was accompanied by task-specific changes in reflex coordination across muscles, a question that cannot be addressed using single-joint perturbations. Reflexes were elicited during the maintenance of posture by perturbing the arm with a three degrees of freedom robot, configured to have isotropic stiffness of either 10 N/m (compliant) or 10 kN/m (stiff). Perturbation characteristics were matched in both environments. Reflex magnitude was quantified by the average rectified electromyogram, recorded from eight muscles crossing the elbow and shoulder. Reflex coordination was assessed using independent components analysis to compare reflex activation patterns during interactions with stiff and compliant environments. Stretch reflex sensitivity increased significantly in all muscles during interactions with the compliant environment and these changes were not due to changes in background muscle activity. However, there was no significant difference in the reflex coordination patterns observed during interactions with the stiff and compliant environments. These results suggest that reflex modulation occurred through altered use of fixed muscle coordination patterns rather than through a change in reflex coordination. PMID- 18287551 TI - cGMP activates a pH-sensitive leak K+ current in the presumed cholinergic neuron of basal forebrain. AB - In an earlier study, we demonstrated that nitric oxide (NO) causes the long lasting membrane hyperpolarization in the presumed basal forebrain cholinergic (BFC) neurons by cGMP-PKG-dependent activation of leak K+ currents in slice preparations. In the present study, we investigated the ionic mechanisms underlying the long-lasting membrane hyperpolarization with special interest in the pH sensitivity because 8-Br-cGMP-induced K+ current displayed Goldman-Hodgkin Katz rectification characteristic of TWIK-related acid-sensitive K+ (TASK) channels. When examined with the ramp command pulse depolarizing from -130 to -40 mV, the presumed BFC neurons displayed a pH-sensitive leak K+ current that was larger in response to pH decrease from 8.3 to 7.3 than in response to pH decrease from 7.3 to 6.3. This K+ current was similar to TASK1 current in its pH sensitivity, whereas it was highly sensitive to Ba(2+), unlike TASK1 current. The 8-Br-cGMP-induced K+ currents in the presumed BFC neurons were almost completely inhibited by lowering external pH to 6.3 as well as by bath application of 100 microM Ba(2+), consistent with the nature of the leak K+ current expressed in the presumed BFC neurons. After 8-Br-cGMP application, the K+ current obtained by pH decrease from 7.3 to 6.3 was larger than that obtained by pH decrease from pH 8.3 to 7.3, contrary to the case seen in the control condition. These observations strongly suggest that 8-Br-cGMP activates a pH- and Ba(2+)-sensitive leak K+ current expressed in the presumed BFC neurons by modulating its pH sensitivity. PMID- 18287552 TI - Responses to binary taste mixtures in the nucleus of the solitary tract: neural coding with firing rate. AB - The contribution of gustation to the perception of food requires an understanding of how neurons represent mixtures of taste qualities. In the periphery, separate groups of fibers, labeled by the stimulus that evokes the best (largest) response, appear to respond to each component of a mixture. In the brain, identification of analogous groups of neurons is hampered by trial-to-trial variability in response magnitude. In addition, convergence of different fiber types onto central neurons may complicate the classification scheme. To investigate these issues, electrophysiological responses to four tastants: sucrose, NaCl, HCl, and quinine, and their binary mixtures were recorded from 56 cells in the nucleus of the solitary tract (NTS, the 1st synapse in the central gustatory pathway) of the anesthetized rat. For 36 of these cells, all 10 stimuli were repeated at least five times (range: 5-23; median = 10). Results showed that 39% of these cells changed their best stimulus across stimulus repetitions, suggesting that response magnitude (firing rate) on any given trial produces an ambiguous message. Averaged across replicate trials, mixture responses most often approximated the response to the more effective component of the mixture. Cells that responded best to a taste mixture rather than any single-component tastant were identified. These cells were more broadly tuned than were cells that responded best to single-component stimuli and showed evidence of convergence from more than one best stimulus fiber type. Functionally, mixture-best cells may amplify the neural signal produced by unique configurations of basic taste qualities. PMID- 18287553 TI - Role of interneuron diversity in the cortical microcircuit for attention. AB - Receptive fields of neurons in cortical area V4 are large enough to fit multiple stimuli, making V4 the ideal place to study the effects of selective attention at the single-neuron level. Experiments have revealed evidence for stimulus competition and have characterized the effect thereon of spatial and feature based attention. We developed a biophysical model with spiking neurons and conductance-based synapses. To account for the comprehensive set of experimental results, it was necessary to include in the model, in addition to regular spiking excitatory (E) cells, two types of interneurons: feedforward interneurons (FFI) and top-down interneurons (TDI). Feature-based attention was mediated by a projection of the TDI to the FFI, stimulus competition was mediated by a cross columnar excitatory connection to the FFI, whereas spatial attention was mediated by an increase in activity of the feedforward inputs from cortical area V2. The model predicts that spatial attention increases the FFI firing rate, whereas feature-based attention decreases the FFI firing rate and increases the TDI firing rate. During strong stimulus competition, the E cells were synchronous in the beta frequency range (15-35 Hz), but with feature-based attention, they became synchronous in the gamma frequency range (35-50 Hz). We propose that the FFI correspond to fast-spiking, parvalbumin-positive basket cells and that the TDI correspond to cells with a double-bouquet morphology that are immunoreactive to calbindin or calretinin. Taken together, the model results provide an experimentally testable hypothesis for the behavior of two interneuron types under attentional modulation. PMID- 18287554 TI - Frequency modulation during song in a suboscine does not require vocal muscles. AB - The physiology of sound production in suboscines is poorly investigated. Suboscines are thought to develop song innately unlike the closely related oscines. Comparing phonatory mechanisms might therefore provide interesting insight into the evolution of vocal learning. Here we investigate sound production and control of sound frequency in the Great Kiskadee (Pitangus sulfuratus) by recording air sac pressure and vocalizations during spontaneously generated song. In all the songs and calls recorded, the modulations of the fundamental frequency are highly correlated to air sac pressure. To test whether this relationship reflects frequency control by changing respiratory activity or indicates synchronized vocal control, we denervated the syringeal muscles by bilateral resection of the tracheosyringeal nerve. After denervation, the strong correlation between fundamental frequency and air sac pressure patterns remained unchanged. A single linear regression relates sound frequency to air sac pressure in the intact and denervated birds. This surprising lack of control by syringeal muscles of frequency in Kiskadees, in strong contrast to songbirds, poses the question of how air sac pressure regulates sound frequency. To explore this question theoretically, we assume a nonlinear restitution force for the oscillating membrane folds in a two mass model of sound production. This nonlinear restitution force is essential to reproduce the frequency modulations of the observed vocalizations. PMID- 18287555 TI - Modeling GABA alterations in schizophrenia: a link between impaired inhibition and altered gamma and beta range auditory entrainment. AB - The disorganized symptoms of schizophrenia, including severely disordered thought patterns, may be indicative of a problem with the construction and maintenance of cell assemblies during sensory processing and attention. The gamma and beta frequency bands (15-70 Hz) are believed relevant to such processing. This paper addresses the results of an experimental examination of the cortical response of 12 schizophrenia patients and 12 control subjects when presented with auditory click-train stimuli in the gamma/beta frequency band during measurement using magnetoencephalography (MEG), as well as earlier work by Kwon et al. These data indicate that control subjects show an increased 40-Hz response to both 20- and 40-Hz stimulation as compared with patients, whereas schizophrenic subjects show a preference for 20-Hz response to the same driving frequencies. In this work, two computational models of the auditory cortex are constructed based on postmortem studies that indicate cortical interneurons in schizophrenic subjects have decreased GAT-1 (a GABA transporter) and GAD(67) (1 of 2 enzymes responsible for GABA synthesis). The models transition from control to schizophrenic frequency response when an extended inhibitory decay time is introduced; this change captures a possible effect of these GABA alterations. Modeling gamma/beta range auditory entrainment in schizophrenia provides insight into how biophysical mechanisms can impact cognitive function. In addition, the study of dynamics that underlie auditory entrainment in schizophrenia may contribute to the understanding of how gamma and beta rhythms impact cognition in general. PMID- 18287556 TI - Sensitivity of inferior colliculus neurons to interaural time differences in the envelope versus the fine structure with bilateral cochlear implants. AB - Bilateral cochlear implantation seeks to improve hearing by taking advantage of the binaural processing of the central auditory system. Cochlear implants typically encode sound in each spectral channel by amplitude modulating (AM) a fixed-rate pulse train, thus interaural time differences (ITD) are only delivered in the envelope. We investigated the ITD sensitivity of inferior colliculus (IC) neurons with sinusoidally AM pulse trains. ITD was introduced independently to the AM and/or carrier pulses to measure the relative efficacy of envelope and fine structure for delivering ITD information. We found that many IC cells are sensitive to ITD in both the envelope (ITD(env)) and fine structure (ITD(fs)) for appropriate modulation frequencies and carrier rates. ITD(env) sensitivity was generally similar to that seen in normal-hearing animals with AM tones. ITD(env) tuning generally improved with increasing modulation frequency up to the maximum modulation frequency that elicited a sustained response in a neuron (tested C20 constitute <10% of the total fatty acyl-CoAs of RAW264.7 cells versus >50% for MCF7 cells, which somewhat astonishingly contain approximately as much C24:0- and C26:0-CoAs as C16:0- and C18:0-CoAs and essentially equal amounts of C26:1- and C18:1-CoAs. This simple and robust method should facilitate the inclusion of this family of compounds in "lipidomics" and "metabolomics" studies. PMID- 18287619 TI - SAGE and LongSAGE. AB - Serial analysis of gene expression (SAGE) is a high-throughput method for global gene expression analysis that allows the quantitative and simultaneous analysis of a large number of transcripts. SAGE is a digital method and its sensitivity depends only on the number of tags sequenced. Furthermore, SAGE is a powerful tool for finding novel genes that are expressed under certain conditions or in certain tissues. SAGE has been widely used in fields as diverse as cancer research and the development and study of microorganisms. The SAGE method is a series of routine molecular biology procedure and can, at least in principle, be carried out in any laboratory. However, the number of consecutive steps is quite large and in practice, SAGE has been difficult to carry out on a routine basis. PMID- 18287620 TI - Robust-LongSAGE (RL-SAGE): an improved LongSAGE method for high-throughput transcriptome analysis. AB - Serial analysis of gene expression (SAGE) is a powerful technique for large-scale transcriptome analysis in eukaryotes. However, technical difficulties in the SAGE library construction, such as low concatemer cloning efficiency, small concatemer size, and a high level of empty clones, has prohibited its widespread use as a routine technique for expression profiling in many laboratories. We recently improved the LongSAGE library construction method considerably and developed a modified version called Robust-LongSAGE, or RL-SAGE. In RL-SAGE, concatemer cloning efficiency and clone insert size were increased significantly. About 20 PCR reactions are sufficient to make a library with more than 150,000 clones. Using RL-SAGE, we have made 10 libraries of rice, maize, and the rice blast fungus Magnaporthe grisea. PMID- 18287621 TI - aRNA-LongSAGE: SAGE with antisense RNA. AB - In order to generate serial analysis of gene expression (SAGE) libraries from very small samples such as microdissected cells, the starting material must first be amplified via PCR or linear amplification of RNA. In microarray experiments, it has been shown that linear amplification of RNA can be used to generate reliable gene expression profiles and leads to the detection of expression differences that are not seen with nonamplified starting material. As the product of the amplification is amplified antisense RNA (aRNA), linear amplification of RNA cannot be used in combination with the conventional SAGE protocol. The aRNA LongSAGE protocol described herein is an adaptation of the MicroSAGE protocol to the use of aRNA as starting material. PMID- 18287622 TI - SuperSAGE. AB - As a tool for high-throughput, quantitative gene expression analysis, serial analysis of gene expression (SAGE) is one of the most powerful techniques. However, the short size of tags (14 bp) has hindered the application of SAGE to a vast majority of eukaryotes without sufficient genomic resources, including expressed sequence tag and genome sequences. To overcome this problem, we developed SuperSAGE, which is based on 26-bp tags from complementary DNA (cDNA), using EcoP15I as a tagging enzyme. Because longer cDNA fragments can easily be recovered by 3'-rapid amplification of cDNA ends (RACE) PCR using primers corresponding to the 26-bp tag sequences in non-model organisms, SuperSAGE allows the identification of novel genes in all eukaryotic organisms, and recommends itself as a useful platform in various fields of biological studies. Here, we present an updated SuperSAGE protocol, which incorporates several modifications and some recommendations to avoid total failure, particularly in the EcoP15I digestion step. PMID- 18287623 TI - Low-cost-medium throughput Sanger dideoxy sequencing. AB - Serial analysis of gene expression (SAGE) requires the sequencing of DNA. The principal cost of SAGE is largely determined by the cost of sequencing. Therefore, it is important to have access to a robust and affordable sequencing system. Here, we describe such a system based on the sequencing of amplified inserts of concatemer-containing clones. PMID- 18287624 TI - DeepSAGE: higher sensitivity and multiplexing of samples using a simpler experimental protocol. AB - Combining serial analysis of gene expression (SAGE) with pyrophosphatase-based ultra-high-throughput DNA sequencing provides increased sensitivity and cost effective gene expression profiling. The combined techniques obviate the formation and cloning of concatemers and the tedious picking and preparation of sequence templates from bacterial clones that are necessary with SAGE alone. Furthermore, multiplexing of samples or replicates of analysis is included in the experimental design. PMID- 18287625 TI - High-resolution, genome-wide mapping of chromatin modifications by GMAT. AB - One major postgenomic challenge is to characterize the epigenomes that control genome functions. The epigenomes are mainly defined by the specific association of nonhistone proteins with chromatin and the covalent modifications of chromatin, including DNA methylation and posttranslational histone modifications. The in vivo protein-binding and chromatin-modification patterns can be revealed by the chromatin immunoprecipitation assay (ChIP). By combining the ChIP assays and the serial analysis of gene expression (SAGE) protocols, we have developed an unbiased and high-resolution genome-wide mapping technique (GMAT) to determine the genome-wide protein-targeting and chromatin-modification patterns. GMAT has been successfully applied to mapping the target sites of the histone acetyltransferase, Gcn5p, in yeast and to the discovery of the histone acetylation islands as an epigenetic mark for functional regulatory elements in the human genome. PMID- 18287626 TI - 5'- and 3'-RACE from LongSAGE tags. AB - Serial analysis of gene expression (SAGE) studies often yield numerous tags that cannot be mapped to known gene sequences. Intriguingly, these may represent unknown genes, unknown parts of genes, or transcript variants. In order to elucidate the origin of these tags, 3'- and 5'-rapid amplification of complementary DNA ends (RACE) reactions can be performed using primers identical or complementary to SAGE tags. This way, transcript fragments, or indeed the entire uncharacterized transcript, can be cloned and sequenced. PMID- 18287627 TI - Extraction and annotation of SAGE tags using sequence quality values. AB - Data analysis of serial analysis of gene expression (SAGE) tag experiments begins with the extraction of tags from single-pass sequence files of ditag concatemers. When using DNA base quality values generated during base calling, it is possible to control the false-positive discovery rate of unique tags. This chapter describes how to set up a system for generating tag lists from quality associated sequence data. PMID- 18287628 TI - Correction of technology-related artifacts in serial analysis of gene expression. AB - Serial analysis of gene expression (SAGE) is a powerful technique for measuring global gene expression through sampling of transcript tags. SAGE tag collections or libraries serve as a rich data source for differential gene expression analysis, transcriptome mapping, and gene discovery. Transcriptome mapping and gene discovery are facilitated by extensions of SAGE, e.g., Long SAGE, where the transcript tags are elongated by utilization of a different tagging enzyme. SAGE, as a sequencing-based technique, is prone to errors resulting in artifact SAGE tag sequences and erroneous tag numbers. A methodology to pinpoint and correct tag artifacts is necessary to fully exploit the value of large SAGE libraries. SAGEScreen is a tag sequence correction algorithm. The algorithm is a multistep procedure that addresses error rates and performs ditag and tag processing. The error rate estimates are based on a stochastic model of PCR and sequencing related mutations. The ditag processing step is essential for calculation of unbiased tag numbers, and the tag processing step allows for filtration of tag sequence artifacts and adjustment of tag numbers. PMID- 18287629 TI - Duplicate ditag analysis in LongSAGE. AB - The Long serial analysis of gene expression (SAGE) protocol generates ditags from tags with overlapping overhangs, thereby increasing the probability of duplicate ditag formation in LongSAGE. In this chapter, a tool is presented that facilitates the analysis of duplicate ditags in LongSAGE studies to determine whether they should be included or not. PMID- 18287630 TI - Statistical comparison of two or more SAGE libraries: one tag at a time. AB - Several statistical tests have been introduced for the comparison of serial analysis of gene expression (SAGE) libraries to quantitatively analyze the differential expression of genes. As each SAGE library is only one measurement, the necessary information on biological variation or experimental precision is lacking. Therefore, each test includes its own approach to derive such a variance measure from the data set or a theoretical distribution. Because the confidence in tag counts depends on the library size, a test between two or more libraries should be based on original tag counts. When groups of libraries are compared, the test should determine that the proportion of a specific tag in all libraries is the same (null hypothesis), but also offer the possibility to detect specific differences between individual libraries and groups of libraries. The Z-test and the G-test encompass these characteristics and are described for the comparison of two libraries and (two or more) groups of libraries, respectively. PMID- 18287631 TI - Scaling of gene expression data allowing the comparison of different gene expression platforms. AB - Serial analysis of gene expression (SAGE) and microarrays have found a widespread application, but much ambiguity exists regarding the amalgamation of the data resulting from these technologies. Cross-platform utilization of gene expression data from the SAGE and microarray technology could reduce the need for duplicate experiments and facilitate a more extensive exchange of data within the research community. This requires a measure for the correspondence of the results from different gene expression platforms. To date, a number of cross-platform evaluations (including a few studies using SAGE and Affymetrix GeneChips) have been conducted showing a variable, but overall low, concordance using different overall correlation approaches, such as Up/Down classification, contingency tables, and correlation coefficients. Here, we demonstrate an approach to compare two platforms based on the calculation of the difference between expression ratios observed in each platform for each individual transcript. This approach results in a concordance measure per gene, as opposed to the commonly used overall concordance measures between platforms. This between-ratio difference is a filtering-independent measure for between-platform concordance. Moreover, the between-ratio difference per gene can be used to identify transcripts with similar regulation on both platforms. PMID- 18287632 TI - Clustering analysis of SAGE transcription profiles using a Poisson approach. AB - To gain insights into the biological function and relevance of genes using serial analysis of gene expression (SAGE) transcription profiles, one essential method is to perform clustering analysis on genes. A successful clustering analysis depends on the use of effective distance or similarity measures. For this purpose, by considering the specific properties of SAGE technology, we modeled the SAGE data by Poisson statistics and developed two Poisson-based measures to assess similarity of gene expression profiles. By employing these two distances into a K-means clustering procedure, we further developed a software package to perform clustering analysis on SAGE data. The software implementing our Poisson based algorithms can be downloaded from http://genome.dfci.harvard.edu/sager. Our algorithm is guaranteed to converge to a local maximum when Poisson likelihood based measure is used. The results from simulation and experimental mouse retina data demonstrate that the Poisson-based distances are more appropriate and reliable for analyzing SAGE data compared to other commonly used distances or similarity measures. PMID- 18287633 TI - Identifying nonspecific SAGE tags by context of gene expression. AB - Many serial analysis of gene expression (SAGE) tags can be matched to multiple genes, leading to difficulty in SAGE data interpretation and analysis. As only a subset of genes in the human genome are transcribed in a certain type of tissue/cell, we used microarray expression data from different tissue types to define contexts of gene expression and to annotate SAGE tags collected from the same or similar tissue sources. To predict the original transcript contributing a nonspecific SAGE tag collected from a particular tissue, we ranked the corresponding genes by their expression levels determined by microarray. We developed a tissue-specific SAGE tag annotation database based on microarray data collected from 73 normal human tissues and 18 cancer tissues and cell lines. The database can be queried online at: http://www.basic.northwestern.edu/SAGE/. The accuracy of this database was confirmed by experimental data. PMID- 18287634 TI - Methodologies for staining and visualisation of beta-galactosidase in mouse embryos and tissues. AB - This chapter provides information on beta-galactosidase staining of whole mouse embryos, organs, tissue sections, and cultured cells, as well as double staining with horseradish peroxidase and use as a tool for genotyping. Using these protocols, localization of beta-galactosidase can be visualized throughout development and in adult tissues. beta-Galactosidase staining may be used purely as a marker of gene expression and also as a tracer in cell lineage studies. PMID- 18287635 TI - Immunohistochemical detection of beta-galactosidase or green fluorescent protein on tissue sections. AB - With the recent advances in mouse genetics, it is now possible to mark specific cell types genetically in vivo and to study the fate of cells during development and adulthood. Cells are labeled and followed in vivo through the stable expression of reporter genes in particular cell types. The two most commonly used reporter genes are LacZ, which encodes the enzyme beta-galactosidase (beta-gal), and green fluorescent protein (GFP). beta-Gal expression can be detected enzymatically, using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) as a substrate, and GFP can be directly visualized by fluorescence microscopy. However, with single detection of beta-gal or GFP, it is often impossible to determine whether expression of the reporter protein is restricted to a particular cell type. To ascertain the identity of individual cells within a multicellular tissue, beta-gal or GFP proteins must be visualized in conjunction with additional cellular markers. For such experiments, specific antibodies raised against beta-gal or GFP can be used in coimmunofluorescence analyses. Such double-staining analyses on tissue sections are a powerful tool to study transgene expression or to trace cells in multicellular tissues. PMID- 18287636 TI - Detection of reporter gene expression in murine airways. AB - We have shown that to overcome the low levels of expression from gene transfer vector-mediated beta-galactosidase expression in lung, it is essential to replace the cytoplasmic beta-galactosidase gene with a nuclear targeted beta galactosidase gene. We found that lung should be sectioned and fixed prior to staining for beta-galactosidase expression and that en bloc staining of intact lung is inefficient at staining positively transduced cells located deeper in the lung spaces. For GFP fluorescence, it is important to inflate the lungs with fixative prior to freezing and subsequent sectioning. For processing of nasal tissues for beta-galactosidase expression, we expand on a protocol used in previously reported gene transfer studies. PMID- 18287637 TI - Three-dimensional analysis of molecular signals with episcopic imaging techniques. AB - This chapter describes two episcopic imaging methods, episcopic fluorescence image capturing (EFIC) and high-resolution episcopic microscopy (HREM). These allow analysis of molecular signals in a wide variety of biological samples such as tissues or embryos, in their precise anatomical and histological context. Both methods are designed to work with histologically prepared and whole-mount stained material, and both provide high-resolution data sets that lend themselves to 3D visualization and modeling. Specimens are embedded in wax (EFIC) or resin (HREM) and sectioned on a microtome. During the sectioning process, a series of digital images of each freshly cut block surface is captured, using a microscope and CCD camera aligned with the position at which the microtome block holder comes to rest after each cutting cycle. The resulting stacks of serial images retain virtually exact alignment and are readily converted to volume data sets. The two methods differ in how tissue architecture is visualized and hence how specific molecular signals are detected. EFIC uses endogenous, broad-range, tissue autofluorescence to reveal specimen structure. Addition of dyes to the wax embedding medium suppresses detection of any signal except that originating from the block surface. EFIC can be used to detect specific signals (such as LacZ) by virtue of their ability to suppress such fluorescence. In contrast, the plastic embedding medium used in HREM is strongly fluorescent, and tissue architecture is detected at the surface because of the ability of cellular and subcellular structures to suppress this signal. Specific signals generated as a result of chromogenic reactions can be visualized using band-pass filters that suppress the appearance of morphological data. In both methods, the digital volume data show high contrast; for HREM, such data achieve true cellular resolution. Their intrinsic alignment greatly facilitates their use for 3D analysis of transgene activity that can be visualized in the context of complex cellular and tissue morphology. Both methods are relatively simple and can be set up using common laboratory apparatuses. Together, they provide powerful tools for analyzing gene function in embryogenesis or tissue remodeling and for investigating developmental malformations. PMID- 18287638 TI - Fluorescent proteins for cell biology. AB - Through the use of exogenous labels, such as antibodies and synthetic fluorophores, experimenters have been able to readily observe the localization of proteins and organelles within a cell by fluorescence microscopy. The discovery and application of fluorescent proteins spanning a large wavelength range have revolutionized these studies. This chapter attempts to introduce the vast array of these molecules, discuss their characteristics, and assess the advantages and disadvantages that each displays for use in imaging. PMID- 18287640 TI - Autofluorescent proteins for flow cytometry. AB - The unique spectral properties and versatility of autofluorescent proteins have facilitated their widespread use in flow cytometric applications. The ability to analyze heterologous fluorescent protein expression conveniently and noninvasively by individually interrogating cells has facilitated increasingly more sophisticated experimental designs to address important biological questions. Improved multilaser flow cytometers have allowed the fluorescent protein field to flourish by permitting high-speed, multiparametric analysis of biological samples. Fluorescent proteins are well suited for either transient or stable expression analysis. Therefore, achieving efficient gene transfer and expression in cells by transfection or viral transduction is paramount to the optimal use of fluorescent proteins in flow cytometry. The archetypal autofluorescent protein, enhanced green fluorescent protein (eGFP), can be used successfully in combination with other fluorescent protein variants. Two such variants, Cerianthus sp. orange fluorescent protein (cOFP) and a fast maturing variant of Discosoma sp. red protein (DsREDExpress), are well suited for flow cytometric applications in combination with eGFP and do not require special filters for optimal excitation and detection. PMID- 18287639 TI - Detection of GFP during nervous system development in Drosophila melanogaster. AB - Using the vital marker GFP and its spectral variants, it is possible to visualize multiple proteins in individual cells and thereby monitor embryonic development on a cellular and molecular level. In the following chapter we describe how to prepare Drosophila embryos or larvae for live imaging or immunohistochemical staining and provide some guidelines for optimal GFP detection. PMID- 18287641 TI - Fluorescent protein reporter systems for single-cell measurements. AB - The unraveling of the complex dynamic networks that underlie cellular (and, by extension, tissue, organ, and organism) function requires sophisticated mathematical models and, in order to test those models, rich data sets. In addition, even in clonal populations of cells, there is a wide range of variability in cellular function at any given time, even in simple parameters such as the concentration of critical signaling components such as receptors or transcription factors. It remains a matter of conjecture as to whether this is noise, to which the system is inherently robust, or whether the cellular control network can exist in multiple discrete internal states, with indistinguishable input/output characteristics. Fluorescent protein-based methods have two features useful for addressing these issues. First, they can be used to retrieve data from individual cells. Second, in combination with confocal fluorescence microscopy, they can be used nondestructively and can thus follow one or more individual cells in culture or in an intact organism over time. PMID- 18287642 TI - Subcellular imaging of cancer cells in live mice. AB - Dual-color fluorescent cells, with one color in the nucleus and the other in the cytoplasm, enable real-time nuclear-cytoplasmic dynamics to be visualized in living cells in vivo as well as in vitro. To obtain the dual-color cells, red fluorescent protein (RFP) is expressed in the cytoplasm of cancer cells, and green fluorescent protein (GFP) linked to histone H2B is expressed in the nucleus. Nuclear GFP expression allows visualization of nuclear dynamics, whereas simultaneous cytoplasmic RFP expression allows visualization of nuclear cytoplasmic ratios as well as simultaneous cell and nuclear shape changes. This methodology has allowed us to show that the cells and nuclei of cancer cells in the capillaries elongate to fit the width of these vessels. The average length of the major axis of the cancer cells in the capillaries increased to approximately four times their normal length. The nuclei increased their length 1.6 times. Cancer cells in capillaries over 8 microm in diameter were shown to migrate at up to 48.3 microm/h. With the use of dual-color fluorescent cells and the Olympus OV100, a highly sensitive whole-mouse imaging system with both macrooptics and microoptics, it is possible to achieve subcellular real-time imaging of cancer cell trafficking in live mice. Extravasation can also be imaged in real time. Dual-color imaging showed that cytoplasmic processes of cancer cells exited the vessels first, with nuclei following along the cytoplasmic projections. Dual color in vivo cellular imaging was also used to visualize trafficking, nuclear cytoplasmic dynamics, and the viability of cancer cells after their injection into the portal vein of mice. PMID- 18287643 TI - Noninvasive imaging of molecular events with bioluminescent reporter genes in living subjects. AB - Bioluminescence imaging has become a very popular tool for noninvasive monitoring of fundamental biological and molecular processes in small living subjects. Luciferases are light-emitting enzymes that can generate light (known as bioluminescence) after reacting with specific substrates. The emitted light is used as a detection system for luciferase activity, which acts as a "reporter" for the activity of any regulatory elements that control its expression. These enzymes are isolated from various organisms, conveniently modified for expression in mammalian cells, and are extensively used in molecular biology and cell culture experiments. Recent advances in optical technology have opened a new dimension for in vivo application of luciferase enzymes in biomedical research. The most commonly utilized luciferases for in vivo bioluminescence are isolated from two very different sources: firefly luciferase (or beetle luciferase) and renilla luciferase (isolated from sea pansy). Although both these luciferases can produce light following interaction with the substrates, structurally and biochemically they are very different. Here we describe the methods and applications of firefly and renilla luciferases in molecular imaging using small animals. PMID- 18287644 TI - Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation. AB - Green fluorescent protein (GFP) and its variants, small, highly soluble proteins, are routinely used as reporters for patterns of gene expression and the origin of cells in transplantation experiments. When not linked as fusion proteins to other polypeptides, they distribute rapidly in the cytoplasm of a given cell, thus allowing real-time observations on living material. For histological analysis, previous bath fixation of whole organs or tissues seemed obligatory, because, during drop fixation of sections, GFP rapidly leaks from cells whose membrane has been damaged by freezing and/or sectioning. The fluorescence of GFP and its derivatives is retained upon fixation, but most enzyme and antigenic activities of interest will be lost in the whole sample as a consequence of formaldehyde (FA) fixation. We have therefore developed an alternative method to fix GFP in frozen tissue sections by FA vapor. This method prevents leakage and redistribution of GFP and allows any cytochemical method to be applied to unfixed adjacent serial sections. PMID- 18287645 TI - Preparation of avidin conjugates. AB - The high-affinity avidin-biotin system has found applications in different fields of biotechnology, including immunoassays, histochemistry, affinity chromatography, and drug delivery, to name a few. A brief description of avidin and avidin-like molecules, streptavidin, deglycosylated avidin, and NeutraLite avidin is presented in the Chapter 2. With four biotin-binding sites per molecule, the avidin family of proteins is capable of forming tight complexes with one or more biotinylated compounds (1). Typically, the avidin-biotin system is used to prepare signal-amplifying "sandwich" complexes between specificity reagents (e.g., antibodies) and detection reagents (e.g., fluorophores, enzymes, and so on). The specificity and detection reagents are independently conjugated, one with avidin and the other with biotin, or both with biotin, providing synthetic flexibility (2). Avidin conjugates of a wide range of fluorophores, phycobiliproteins, seconday antibodies, microspheres, ferritin, and enzymes commonly used in immunochemistry are available at reasonable prices, making their small-scale preparation impractical and not cost effective (see Note 1). However, conjugations of avidin to specific antibodies, to uncommon enzymes, and to other proteins and peptides are often performed on-site. A general protocol for the conjugation of avidin to enzymes, antibodies, and other proteins is described in this chapter. PMID- 18287646 TI - Coupling of antibodies with biotin. AB - The avidin-biotin bond is the strongest known biological interaction between a ligand and a protein (Kd = 1.3 x 10-15 M at pH 5.0) (1). The affinity is so high that the avidin-biotin complex is extremely resistant to any type of denaturing agent (2). Biotin (see Fig. 1) is a small, hydrophobic molecule that functions as a coenzyme of carboxylases (3). It is present in all living cells. Avidin is a tetrameric glycoprotein of 66,000-68,000 molecular weight, found in egg albumin and in avian tissues. The interaction between avidin and biotin occurs rapidly, and the stability of the complex has prompted its use for in situ attachment of labels in a broad variety of applications, including immunoassays, DNA hybridization (4-6), and localization of antigens in cells and tissues (7). Avidin has an isoelectric point of 10.5. Because of its positively charged residues and its oligosaccharide component, consisting mostly of mannose and glucosamine (8), avidin can interact nonspecifically with negative charges on cell surfaces and nucleic acids, or with membrane sugar receptors. At times, this causes background problems in histochemical and cytochemical applications. Streptavidin, a near-neutral, biotin-binding protein (9) isolated from the culture medium of Streptomyces avidinii, is a tetrameric nonglycosylated analog of avidin with a molecular weight of about 60,000. Like avidin, each molecule of streptavidin binds four molecules of biotin, with a similar dissociation constant. The two proteins have about 33% sequence homology, and tryptophan residues seem to be involved in their biotin-binding sites (10,11). In general, streptavidin gives less background problems than avidin. This protein, however, contains a tripeptide sequence Arg-Tyr-Asp (RYD) that apparently mimics the binding sequence of fibronectin Arg-Gly-Asp (RGD), a universal recognition domain of the extracellular matrix that specifically promotes cell adhesion. Consequently, the streptavidin-cell-surface interaction causes high background in certain applications (12). PMID- 18287647 TI - Optimization of detection and quantification of proteins on membranes in very high and very low abundance using avidin and streptavidin. AB - Numerous methods have been published for the detection of protein using avidin biotin technology. Complications can arise using this system when the protein of interest is in extremely high or low abundance. The ability to successfully detect high- or low-abundance proteins is dependent on the detection system selected. The expression of endogenous biotinylated proteins in rat liver homogenate will be used to illustrate the methods utilized for the detection of high- and low-abundance proteins on a membrane. The advantages and disadvantages of enzymatic and direct fluorescence detection systems are discussed. PMID- 18287648 TI - Use of the avidin (imino)biotin system as a general approach to affinity precipitation. AB - Biospecific interactions are used in many capturing and bioseparation steps. A typical situation is the coupling of a biospecific ligand to a chromatographic stationary phase for affinity chromatography. This approach has two possible drawbacks. The first is that a chromatographic column may be awkward to use in experimental setups; the second is related to the need to develop a dedicated coupling chemistry for any given affinity ligand. In affinity precipitation, the biospecific affinity ligand is instead linked to a stimuli-responsive molecule to yield a so-called affinity macroligand (AML). Upon stimulation, such molecules show abrupt yet reversible precipitation from aqueous solution. Capture by affinity precipitation just requires the addition of the stimuli-responsive AML to the raw target solution followed by selective precipitation of the formed affinity complex via the application of the stimulus. The need for the synthesis of a dedicated AML may be circumvented by the use of an avidin-activated stimuli responsive precursor, to which any biotinylated affinity ligand can be securely linked via the well-known strong avidin-biotin interaction. PMID- 18287649 TI - Capturing biotinylated proteins and peptides by avidin functional affinity electrophoresis. AB - Avidin functional affinity electrophoresis (AFAEP) is a variational method of affinity electrophoresis. In this technique, avidin is immobilized within a small area of the gel matrix by interaction with acrylamide and/or polyacrylamide either directly or through bifunctional linker glutaraldehyde during polymerization. Analytes can be heated with Tris-glycine sodium dodecyl sulfate (SDS) sample buffer so that biotinylated peptides/proteins are negatively charged and migrate electrophoretically towards the cathode through the avidin zone regardless of their isoelectric point (pI) values. Alternatively, if the behavior of the biotinylated analytes is known, the SDS treatment is not required. The polarity of the electrodes is set such that biotinylated analytes migrate electrophoretically through the avidin zone. This technique can work with or without SDS in gel running buffer. The AFAEP method allows the capture and concentration of biotinylated peptides/proteins. The values of this technique stem from a combination of merits of polyacrylamide gel electrophoresis and affinity technology. PMID- 18287650 TI - Functionality screen of streptavidin mutants by non-denaturing SDS-PAGE using biotin-4-fluorescein. AB - Site-directed mutagenesis or directed evolution of proteins often leads to the production of inactive mutants. For streptavidin and related proteins, mutations may lead to the loss of their biotin-binding properties. With high-throughput screening methodologies in mind, it is imperative to detect, prior to the high density protein production, the bacteria that produce non-functional streptavidin isoforms. Based on the incorporation of biotin-4-fluorescein in streptavidin mutants present in Escherichia coli bacterial extracts, we detail a functional screen that allows the identification of biotin-binding streptavidin variants. Bacteria are cultivated in a small volume, followed by a rapid treatment of the cells; biotin-4-fluorescein is added to the bacterial extract and loaded on an Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis (SDS-PAGE) under non denaturing conditions. Revealing is performed using a UV transilluminator. This screen is thus easy to implement, cheap and requires only readily available equipment. PMID- 18287651 TI - Application of biotin-4-fluorescein in homogeneous fluorescence assays for avidin, streptavidin, and biotin or biotin derivatives. AB - Biotin-4-fluorescein (B4F) is a convenient molecular probe for (strept)avidin and for unlabeled biotin in homogeneous fluorescence assays. The primary standard is a 16 microM working solution of d-biotin which is used to titrate an aliquot of a (strept)avidin stock solution while monitoring the tryptophane fluorescence of (strept)avidin. This serves to standardize the (strept)avidin stock solution, an aliquot of which is then titrated with a roughly 16 microM working solution of B4F while monitoring the fluorescence of B4F. Specific binding is accompanied by quenching, but after saturation of all binding sites, the appearance of free ligand causes a sharp rise of intense fluorescence, the beginning of which allows to calculate the effective concentration of B4F in the working solution. Measurement of avidin in a crude sample is exemplified by mixing 8 pmol of B4F with various amounts of diluted egg white in a volume of 1 mL. Hereby, the extent of fluorescence quenching linearly correlates with the concentration of functional avidin. Moreover, a sharp minimum of fluorescence is observed when exactly 2 pmol of avidin is present in the sample. The latter assay has been adapted to measure between 0.5 and 5 pmol of d-biotin in 1 mL of sample by adding 1.9 pmol of avidin and 8 pmol of B4F. This competitive assay correctly measures the small dose of d-biotin in multivitamin tablets (e.g., 150 microg in 5 g of solid) after subtracting the background fluorescence of the colored aqueous solution. PMID- 18287652 TI - Quantitative recovery of biotinylated proteins from streptavidin-based affinity chromatography resins. AB - The strong interaction between streptavidin and biotin is one of the most commonly exploited tools in chemistry and biology. Methods for the facile derivatization of a variety of molecules (in particular, proteins) with biotin have been introduced, in order to allow their efficient recovery, immobilization and detection with streptavidin-based reagents. However, when desired, the release of biotinylated proteins from the streptavidin-based reagents remains a major problem, due to the extraordinary stability of this complex. This chapter presents a protocol developed in our laboratory for the quantitative elution of biotinylated proteins from streptavidin sepharose, featuring harsh elution conditions and competition with free biotin. The usefulness of the method is shown by the recovery of biotinylated proteins from organ homogenates, obtained from mice perfused with a reactive ester derivative of biotin. PMID- 18287653 TI - High-yield production and purification of recombinant T7-tag mature streptavidin in glucose-stressed E. coli. AB - The overexpression of toxic recombinant proteins is often problematic, leading to either low production levels or inclusion bodies. Streptavidin is no exception and thus the highest production level reported to date for streptavidin is 70 mg/L of functional protein. Herein, we report on the production in Escherichia coli and the purification of a recombinant mature streptavidin bearing a T7-tag. Optimization of critical parameters, including the glucose concentration, the pH and the time of induction as well as the use of BL21(DE3)pLysS cell strain, affords up to 120 mg/L functional streptavidin in soluble form. The yield can be further increased by an osmotic stress during the preculture by adding highly concentrated glucose before the inoculation of the culture medium, thus affording reproducibly 230 mg/L of soluble streptavidin. A single denaturing-renaturing step and affinity chromatography afford highly active tetrameric protein with >3.8/4.0 active sites. PMID- 18287654 TI - Endogenous biotin in rat brain: implications for false-positive results with avidin-biotin and streptavidin-biotin techniques. AB - The interaction between avidin and biotin or streptavidin and biotin forms the basis of several widely used immunohistochemical techniques. An assumption inherent to these techniques is that endogenous biotin is not present in the tissue in detectable quantities, as neither avidin nor streptavidin can discriminate between endogenous biotin and biotinylated antibodies. However, biotin is a known cofactor for numerous carboxylases required in oxidative metabolism, raising the possibility for potential false-positive results in many tissues. This issue has been appreciated in liver and kidney tissue, but has received very little attention in nervous tissue. To address this concern, we examined the distribution of biotin throughout the rat central nervous system using avidin- and streptavidin-based detection systems, as well as a monoclonal antibody raised against biotin. Significant levels of endogenous biotin were identified within specific neuronal types, particularly in neurons associated with the cerebellar motor system and the brainstem auditory system. Non-specific (non-biotin) interactions of avidin and streptavidin conjugates with rat brain tissue were further identified and were most pronounced in the lower brainstem. The binding of avidin- and streptavidin-conjugated markers to endogenous biotin and other non-specific interactions with neural tissue were overcome by several methods including the use of blocking kits, prolonged post-fixation of the tissue in paraformaldehyde, or omission of Triton X-100 from the working solution. Without these measures, a reliable estimate of immunolabel may only be achieved in certain brain regions with markers conjugated directly to secondary antibodies. PMID- 18287655 TI - Pseudo-immunolabelling with the avidin-biotin-peroxidase complex due to the presence of endogenous biotin in the retina. AB - Immunodetection techniques are dependent on enzyme-protein conjugates for the visualization of antigen-antibody complexes. One of the most widely used is the avidin-biotin-peroxidase complex (ABC) method. However, treatment of certain tissues with ABC reagents alone may result in high background, which is indicative for the presence of endogenous biotin or biotinylated proteins. In goldfish and salamander retinal sections, we observed a distinct staining pattern, presumably through binding of avidin to endogenous biotin in Muller cells. These findings summon for caution in the application of detection systems based on biotinylated antibodies or biotinylated DNA probes. PMID- 18287656 TI - Use of synthetic peptides for identifying biotinylation sites in human histones. AB - Posttranslational modifications of histones play an important role in the regulation of chromatin structure and, hence, gene regulation. Recently, we have identified a novel modification of histones: binding of the vitamin biotin to lysine residues in histones H2A, H3, and H4. Here, we describe a procedure to identify those amino acids that are targets for biotinylation in histones. Briefly, the following analytical sequence is used to identify biotinylation sites: (i) short peptides (<20 amino acids in length) are synthesized chemically; amino acid sequences in the peptides are based on the sequence in a given region of a given histone; (ii) peptides are incubated with biotinidase or holocarboxylase synthetase to conduct enzymatic biotinylation; and (iii) biotin in peptides are probed using streptavidin peroxidase. Amino acid substitutions (e.g., lysine-to-alanine substitutions) in synthetic peptides can be used to corroborate identification of biotinylation sites. PMID- 18287657 TI - Detection and quantitation of the activity of DNA methyltransferases using a biotin/avidin microplate assay. AB - The biotin-avidin microplate assay is a sensitive method to measure methylation of biotinylated oligonucleotide substrates by DNA methyltransferases (MTases). The methylation reaction is carried out in solution using [methyl-3H]-AdoMet. Afterwards, the oligonucleotides are immobilized on an avidin-coated microplate, where the incorporation of [3H]-labeled methyl groups into the DNA is stopped by addition of unlabeled AdoMet to the binding buffer. Separation of radioactively labeled DNA from unreacted AdoMet and enzyme is performed by washing steps. Subsequently, the radioactivity incorporated into the DNA is released by a nucleolytic digestion of the DNA. By liquid scintillation counting, the amount of DNA methylation can be determined. Advantages of the microplate assay are its high sensitivity which allows the detection of low amounts of DNA methylation, the efficient separation of reaction components resulting in a low background of radioactivity and a high accuracy (+/-10%) and reliability. Furthermore, the assay is very convenient, fast and well suited for automation. PMID- 18287658 TI - Anti-biotin antibodies offer superior organelle-specific labelling of mitochondria over avidin or streptavidin. AB - A number of endogenously biotinylated proteins are found in both cytosol and mitochondria of mammalian cells from many tissues, including liver, spleen, pancreas, kidney, and intestine. Therefore, caution should be taken when using the biotin detection system. Endogenous biotin can interfere with staining systems that employ the use of biotin-avidin- or biotin-streptavidin-based detection systems and may therefore result in high, non-specific background staining. Here, we show that this endogenous biotin reactivity can be deliberately exploited and used as a specific mitochondrial marker in both light and electron microscopy as well as for identifying mitochondrial fractions on Western blot. PMID- 18287659 TI - Mammalian lectin as tool in glycochemistry and histochemistry with relevance for diagnostic procedure. AB - Carbohydrates represent a fundamental building unit of living organisms. Many contemporary results introduce these substances as medium with remarkable data storage capacity (glycocode) that is decoded by special receptor counterpartners, lectins. Animal so-called endogenous lectins are presented here as the biotinylated tools for normal lectin histochemistry in accompany with biotinylated (neo)glycoconjugates employed in reverse lectin histochemistry. Practical lesson how to employ these probes in cell/tissue labeling including multiple cell labeling at the single-cell level is also included. Position of glycocode and endogenous lectins in squamous epithelium biology under physiological condition and after the malignant transformation is shown as example of the employment of mentioned probes in research and diagnostics. PMID- 18287660 TI - A multiplex approach to isotyping antigen-specific antibodies using biotinylated antigen/streptavidin-phycoerythrin. AB - Analytical methods characterizing the immunogenicity of antigens are useful for monitoring, characterizing and predicting antibody responses to therapeutic biologics or vaccines. Distinct Luminex microspheres coupled with protein G, anti human immunoglobulin (Ig)A, anti-human IgM and anti-human IgE were developed for the simultaneous capture of total IgG, IgA, IgM and IgE (IgGAME) antibodies from human or non-human primate serum. The fraction of antigen-specific antibodies captured on the beads was detected using biotinylated antigen/streptavidin phycoerythrin. The method was demonstrated by isotyping antibodies directed against an anti-CD11a antibody therapeutic (RAPTIVA/efalizumab) from the serum of a cynomolgus monkey hyper-immunized with RAPTIVA over a 15-month period. The quantitative range of the antibody measurements, using 5 mul of sample, was determined to be 15 ng/ml to 50 mug/ml in 10% serum. By the use of any biotinylated antigen as a detector, this multiplexed isotyping assay can be broadly applied to human and non-human primate IgGAME immunogenicity studies. PMID- 18287661 TI - Biotin-protein bond: instability and structural modification to provide stability for in vivo applications. AB - Biotinylation of proteins is a powerful tool for investigating biological phenomenon, both in vitro and in vivo. Biotinylating reagents that form covalent bonds with several types of amino acid residues are commercially available. However, most, if not all, of these commercially available biotinylating agents produce biotin-protein bonds that are susceptible to cleavage in human plasma. Here, we describe the use of immunoglobulin G as a model protein for evaluation of biotin-protein bond stability and for the investigation of the mechanism of biotin release. We also describe the synthesis of a biotin-protein bond that is stable in human plasma and a method for evaluation of that stability. PMID- 18287662 TI - Rat carotid artery balloon injury model. AB - Numerous and diverse experimental animal models have been used over the years to examine reactions to various forms of blood vessel disease and/or injury across species and in multiple vascular beds in a cumulative effort to relate these findings to the human condition. In this context, the rat carotid artery balloon injury model is highly characterized and commonly used for investigating gross morphological, cellular, biochemical, and molecular components of the response to experimentally induced arterial injury. The mechanical damage caused by the balloon catheter completely removes the intimal endothelial lining and creates a distending mural injury in the operated vessel. This elicits a reproducible remodeling response characterized by vascular smooth muscle cell (SMC) mitogenesis and migration (through phenotypic switching), SMC apoptosis, partial vascular endothelial cell regeneration, enhanced matrix synthesis, and establishment of an invasive neointima in time-dependent fashion. This multi factorial process allows for investigation of these many important pathophysiological processes and can serve as a valuable "proof-of-concept" tool to verify and substantiate in vitro results; however, inherent anatomical and adaptive constraints of this in vivo model ration comparison to the diseased human system (see Note 1). In this chapter, brief overview of the materials needed and the methodologies commonly employed for successful routine performance of this important experimental animal model is provided. Individual sub-sections will cover animal care and handling, pre-operative and post-operative procedures, and the surgery proper. Protocols for histopathology and morphometry and procedures for data management and interpretation pertinent to the rat carotid artery balloon injury model are discussed in Chapter 2. PMID- 18287663 TI - Histological and morphometric analyses for rat carotid balloon injury model. AB - Experiments aimed at analyzing the response of blood vessels to mechanical injury and ensuing remodeling responses often employ the highly characterized carotid artery balloon injury model in laboratory rats. This approach utilizes luminal insertion of a balloon embolectomy catheter into the common carotid artery with inflation and withdrawal resulting in an injury characterized by vascular endothelial cell (EC) denudation and medial wall distension. The adaptive response to this injury is typified by robust vascular smooth muscle cell (SMC) replication and migration, SMC apoptosis and necrosis, enhanced synthesis and deposition of extracellular matrix (ECM) components, partial vascular EC regeneration from the border zones, luminal narrowing, and establishment of a neointima in time-dependent fashion. Evaluation of these adaptive responses to blood vessel injury can include acute and longer term qualitative and quantitative measures including expression analyses, activity assays, immunostaining for a plethora of factors and signals, and morphometry of neointima formation and gross mural remodeling. This chapter presents a logical continuation of Chapter 1 that offers details for performing the rat carotid artery balloon injury model in a standard laboratory setting by providing commonly used protocols for performing histological and morphometric analyses in such studies. Moreover, procedures, caveats, and considerations included in this chapter are highly relevant for alternative animal vascular physiology/pathophysiology studies and in particular those related to mechanisms of vascular injury and repair. Included in this chapter are specifics for in situ perfusion-fixation, tissue harvesting and processing for both snap-frozen and paraffin-embedded protocols, specimen embedding and sectioning, slide preparation, several standard histological staining steps, and routine morphological assessment. PMID- 18287664 TI - Plaque rupture model in mice. AB - It is widely believed that rupture of a vulnerable atherosclerotic plaque leads to acute coronary events and stroke. However, the exact mechanisms involved in the plaque rupture remain unknown. Pathological animal models are valuable in the research on human disease mechanism, their therapy, and drug development. Recently, we have proposed a novel murine model of atherosclerotic plaque rupture associated with not only intraplaque hemorrhage but also luminal thrombus using combination treatments of ligation and perivascular cuff placement on the carotid artery. PMID- 18287665 TI - Immunostaining of mouse atherosclerotic lesions. AB - Atherosclerotic lesions develop through interactions with diverse cell types whose functions are determined by a complex array of regulators. Immunostaining is now a commonly applied technique to identify these numerous cell types and regulators in lesions. The principle of the technique is that an antibody is incubated with the tissue under conditions that favor a specific interaction with its antigen. This is subsequently visualized most commonly by a chromogenic substrate that produces a colored precipitate at the location of the antigen antibody interaction. When appropriately applied, it is a powerful technique to provide mechanistic insight into the atherogenic process. However, the complexity of atherosclerotic tissue can provide challenges to ensuring that development of the chromogen is due to specific antigen-antibody interaction. Thus, the determination of specific interactions frequently requires the judicious use of appropriate control experiments. PMID- 18287666 TI - Surgical animal model of ventricular hypertrophy. AB - In response to an increased afterload, the myocardium undergoes a complex adaptation by which wall stress is normalized and cardiac output is maintained. Although the consensus suggests that the increase of the myocardial mass is a necessary adaptive process to accommodate the increased workload, there is growing evidence that hypertrophy ultimately results in pathological remodeling and deterioration of cardiac function. Despite intense investigation, our understanding of the cellular mechanisms that are responsible for the initiation and the maintenance of this adaptation is largely incomplete and preventing or regressing left ventricular hypertrophy (LVH) is a major challenge. This chapter provides a detailed description of the procedures necessary to induce LVH by coarctation of the transverse aorta and to analyze the effects of the increased hemodynamic load on cardiac mass, cardiomyocyte size, and cardiac performance. PMID- 18287667 TI - Animal models of hypertension. AB - Hypertension affects approximately 25% of adults and is a major risk factor for cardiovascular disease. Although there are currently adequate therapeutic options for humans with hypertension, the molecular mechanisms underlying hypertension are still relatively unknown. The generation of hypertensive animal models provides an excellent modality to not only study the pathophysiology but also test innovative therapeutics. This chapter describes the detailed methods that utilize the drinking water of rats to develop models of nitric oxide synthase (NOS) inhibition-induced, guanosine triphosphate cyclohydrolase (GTPCH) inhibition-induced, and glucocorticoid-induced hypertension. PMID- 18287668 TI - Rat models of cardiac insulin resistance. AB - Cardiovascular disease is the leading cause of death in the industrialized world. Diabetes is a major risk factor for cardiovascular disease. Unchecked diabetes can also lead to renal failure, blindness, heart attack, stroke, and amputation. The focus of this chapter will be to review the different mechanisms of insulin resistance (IR)/type 2 diabetes and various animal models used to study cardiac changes during these conditions. PMID- 18287669 TI - The isolated, perfused pseudo-working heart model. AB - Increased interest in cardiac safety and renewed interest in drugs for treating myocardial ischemia and congestive heart failure have led to increased use of cardiovascular models. Unfortunately, many molecular or cell-based screens are not perfect predictors of activity in vivo. One rapid means to test "proof of principle" in hearts is the isolated or Langendorff perfused heart system. This allows direct measurement of cardiac contractile function and coronary flow without interference from changes in the systemic circulation. The setup basically is a heart whose coronary arteries are retrogradely perfused through the aortic root with heated and oxygenated buffer solutions. We will describe the constant pressure technique as this is the most commonly used and allows us to directly measure changes in coronary flow without changes in turgor noted with constant flow methods. Instead of a classical working heart preparation, we describe the use of a balloon properly fitted into the left ventricle to measure end diastolic and peak systolic pressure making this a much simpler and less expensive technique without loss of scientific quality. Such a setup can be used to measure effects of compounds or treatments on coronary flow, contractile function either in the normal or pathological states. PMID- 18287670 TI - Altering and analyzing glucose metabolism in perfused hearts of transgenic mice. AB - Glucose metabolism plays an important role in cardiac bioenergetics that changes under various stress conditions including hypertrophy, diabetic cardiomyopathy, and ischemia-reperfusion injury. To understand the role of glycolysis under these conditions, we have altered several steps of the glycolytic pathway specifically in the heart. In this chapter, we describe methods used to produce cardiac targeted transgenic mice and procedures for measuring various glucose metabolites including glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, and glycogen. Also, we describe methods for measuring glucose transport and glycolysis in perfused mouse hearts. Using these methods, we show that mice over expressing cardiac-specific kinase-deficient 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase (Mykd-PFK-2) show reduced glucose transport and reduced glycolysis when compared with control. The metabolites glucose-6-phosphate, fructose-6-phosphate, and glycogen were elevated, whereas fructose-1,6 bisphosphate was reduced in the transgenic Mykd-PFK-2 mouse hearts. PMID- 18287671 TI - Methods in the evaluation of cardiovascular renin angiotensin aldosterone activation and oxidative stress. AB - Renin angiotensin aldosterone system (RAAS) activation plays an essential role in the development of cardiovascular disease (CVD). Multiple pathophysiologic processes are able to activate RAAS, among which hypertension, obesity, diabetes mellitus 2, and chronic kidney disease deserve special attention, because they are the main contributors to CVD. Adding to the well-known effects of RAAS overactivity on the vasculature and water and electrolyte balance, current evidence links abnormal activation of the RAAS to increased production of reactive oxygen species (ROS) and oxidative stress. This association is mediated at least partially through interaction of angiotensin II (Ang II) with its receptor angiotensin receptor 1 (AT1R) in cardiovascular tissue, and subsequent activation of the nicotinamide adenine dinucleotide phosphate (NADPH) enzymatic complex, which finally leads to increased ROS production. This resulting state of enhanced oxidative stress contributes largely to generalized atherosclerosis and finally to CVD. The generation of animal models of increased RAAS and Ang II expression, in particular the Ren2 rodent model, provides important opportunities to better characterize the relationship between this system and the production of ROS. This chapter describes methods to evaluate, characterize, and quantify the activity of the RAAS and NADPH oxidase, as well as the production of ROS production in animal model of RAAS. PMID- 18287672 TI - Detection and quantification of apoptosis in the vasculature. AB - The integral role of apoptosis in the pathogenesis of cardiovascular diseases has been extensively studied and characterized in recent years. The study of cell death in the vasculature has significantly contributed to our knowledge of vascular disease pathology and has played a role in identifying potential therapeutic strategies for these diseases. This chapter describes a number of standard, widely used protocols for detecting and quantifying apoptosis in vessel wall cells and tissue. These techniques include terminal deoxynucleotidyl transferase dUTP nick-end labeling staining for DNA fragmentation, Hoechst staining for chromatin condensation, Annexin V staining, labeling for phosphatidylserine externalization, Western blot assessment of caspase cleavage, immunofluorescence detection of caspase activation, assessment of mitochondrial membrane depolarization and cytochrome c release, and a splenocyte assay for quantifying susceptibility to immune cell-mediated apoptosis. PMID- 18287673 TI - Endothelial progenitor cell and mesenchymal stem cell isolation, characterization, viral transduction. AB - Endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) have emerged as potentially useful substrates for neovascularization and tissue repair and bioengineering. EPCs are a heterogeneous group of endothelial cell precursors originating in the hematopoietic compartment of the bone marrow. MSCs are a rare population of fibroblast-like cells derived from the bone marrow stroma, constituting approximately 0.001-0.01% of the nucleated cells in the marrow. Both cells types have been isolated from the bone marrow. In addition, EPC can be isolated from peripheral blood as well as the spleen, and MSC has also been isolated from peripheral adipose tissue. Several approaches have been used for the isolation of EPC and MSC, including density centrifugation and magnetic bead selection. Phenotypic characterization of both cell types is carried out using immunohistochemical detection and fluorescence-activated cell sorting analysis of cell-surface molecule expression. However, the lack of specific markers for each cell type renders their characterization difficult and ambiguous. In this chapter, we describe the methods that we use routinely for isolation, characterization, and genetic modification of EPC and MSC from human, rabbit, and mouse peripheral blood and bone marrow. PMID- 18287674 TI - Assessment of endothelial damage/dysfunction: a focus on circulating endothelial cells. AB - Endothelial injury represents a major initiating step in the pathogenesis of vascular disease and atherosclerosis. The identification and quantification of circulating endothelial cells (CECs) has evolved as a novel marker of endothelial function. As a technique, it correlates with other markers of endothelial function such as flow-mediated dilation, the measurement of von Willebrand factor, and tissue plasminogen activator. Quantification of CECs is difficult due to low numbers, variable morphology, and a lack of standardization in current techniques used. CECs appear to be a different population of cells to endothelial progenitor cells. Increased CECs have been noted in a number of disease states and is evolving as a novel method of assessment of both disease severity and response to treatment. PMID- 18287675 TI - Evaluation of endothelial function by in vivo microscopy. AB - This chapter describes a method that permits simultaneous measurement of leukocyte- endothelium interactions and endothelial nitric oxide (NO) levels in the microcirculation in vivo. The method is also useful to study the effect of NO replenishing therapy on adhesion of leukocytes to the vascular endothelium in acute and chronic inflammatory states of the cardiovascular system. This research approach requires the combination of two well-established physiology techniques, that is, intravital microscopy and real-time measurement of NO with microelectrodes. Intravital microscopy is considered the method of choice to monitor leukocyte-endothelial cell interactions in intact vascular beds of live animals. In vivo microscopy is currently used to study the endothelial cell phenotype of mice carrying mutations or deletion of targeted genes. Intravital microscopy is also used to study endothelial cell function in acute (e.g., ischemia-reperfusion injury) and chronic (e.g., hypercholesterolemia, hyperglycemia, and diabetes) inflammatory states of the cardiovascular system. NO sensors allow for continuous, amperometric quantification of NO levels in cells and organ tissues. Coupling of NO electrode technology with intravital microscopy has recently permitted to measure NO bioavailability in the normal and inflamed microcirculation. The method described here can be used to study in vivo how acute and chronic inflammatory states of the cardiovascular system alter endothelial function resulting in endothelial cell activation and damage. PMID- 18287676 TI - Haptotaxis of endothelial cell migration under flow. AB - Endothelial cell (EC) migration plays an important role in embryonic vasculogenesis, angiogenesis, and wound healing. EC migration can be regulated by extracellular matrix (ECM) and hemodynamic forces through haptotaxis (induced by an ECM gradient) and mechanotaxis (induced by mechanical forces). Previously, the effects of haptotaxis or mechanotaxis alone on EC migration have been studied; however, the dual effect of haptotaxis and mechanotaxis on EC migration is not known. We developed a micropatterning technique to generate step changes of collagen surface density to monitor haptotactic EC migration. To investigate the crosstalk between haptotaxis and mechanotaxis on EC migration, we used an in vitro flow system to apply a well-defined fluid shear stress on ECs cultured on the micropatterned collagen. The study on the effects of haptotaxis and mechanotaxis on EC migration will provide a rational basis for promoting vascular wound healing, angiogenesis, and vascularization in engineered tissues. PMID- 18287677 TI - Isolation, culture, and functional analysis of adult mouse cardiomyocytes. AB - The pathogenesis of heart disease and the development of myocardial failure are highly dependent upon the cardiomyocyte-the basic contractile cell within the heart. Understanding and elucidating the complex networks that regulate cardiomyocyte function are central to the development of specific target-based therapeutic interventions. The relative recent advances in molecular genetics and generation and routine usage of transgenic and knockout mouse models have further necessitated that previously established cardiomyocyte methods be adapted for the isolation, culture, and study of primary adult murine cardiomyocytes, both freshly isolated and in culture. Such adaptation is based not only upon scalability of established techniques, as the mouse heart is an order of magnitude smaller than the rat heart, but also upon properties unique to the mouse. This chapter therefore describes current methods for the isolation, culture, and functional analysis of adult murine cardiomyocytes. PMID- 18287678 TI - Mechanical measurement of contractile function of isolated ventricular myocytes. AB - Isolation of ventricular myocytes from all species of animals has revolutionized the field of cardiovascular research, allowing the assessment of true cardiac effects of drugs, treatments, and so on. With recent advances in physiology at the cellular level, direct assessment of isolated ventricular myocyte mechanics has become an increasingly powerful and important area of research. This technique provides important information on excitation-contraction coupling of the heart, either when investigating the effect of a drug on the heart or mechanical function under certain pathophysiological states. The goal of this chapter is to provide a detailed list of methods for isolation of ventricular myocytes from both rats and mice, as well as a list of certain "tricks" that have proven useful in enhancing the yield of viable cells from the whole heart. Mastering the isolation of myocytes will provide a much better framework for real time beat-to-beat recording of myocyte contraction as well as intracellular Ca2+ transients. PMID- 18287679 TI - Studying ischemia and reperfusion in isolated neonatal rat ventricular myocytes using coverslip hypoxia. AB - In vitro experimental models designed to study the effects of hypoxia and ischemia typically employ oxygen-depleted media and/or hypoxic chambers. These approaches, however, allow for metabolites to diffuse away into a large volume and may not replicate the local buildup of metabolic byproducts that occur in ischemic myocardium in vivo. Coverslip hypoxia (CSH) is a recently described method for studying hypoxia and ischemia derived from the byproducts and metabolites of contractile ventricular myocytes. Hence, this method is dependent on the purity and contractile activity of the isolated myocytes. We describe herein methods for isolating neonatal rat ventricular myocytes with these characteristics, as well as means for performing CSH, identifying viable and compromised myocytes after coverslipping, and tracking pH changes during CSH. PMID- 18287680 TI - Isolation and functional studies of rat aortic smooth muscle cells. AB - Migration, proliferation, and collagen synthesis by vascular smooth muscle cells are thought to be key events involved in the pathogenesis of cardiovascular disease. Following endothelial injury, smooth muscle cells (SMCs) in the intima of the blood vessels assume a synthetic, promitogenic phenotype resulting in their migration, proliferation, and deposition of extracellular matrix within the neointimal tissue. This chapter describes a method of isolation of SMCs from rat aorta and in vitro assays to characterize these abnormal SMC functions. PMID- 18287682 TI - Assessment of protein glycoxidation in ventricular tissues. AB - Advanced glycation end products are permanently modified protein derivatives formed in the presence of reducing sugars, such as glucose, fructose, hexose phosphates, trioses, and triose-phosphates by non-enzymatic glycation and oxidation ("glycoxidation") reactions and further irreversible rearrangements. Numerous studies have revealed the pivotal role of protein glycoxidation in the pathogeneses of diabetes-related and age-related diseases. Protein glycoxidation is generally recognized both as a hallmark and as a promoter for progression of diabetes-related and age-related ailments, particularly in cardiovascular system such as increased vascular and myocardial stiffness, endothelial dysfunction, altered vascular injury responses, and atherosclerotic plaque formation. An appropriate surveillance on abnormal protein glycoxidation at an early stage of disease progression is of clinical and practical importance to handle diabetes related and age-related cardiovascular complications especially those leading to ventricular dysfunction. PMID- 18287681 TI - Detection of reactive oxygen species and nitric oxide in vascular cells and tissues: comparison of sensitivity and specificity. AB - Reactive oxygen and nitrogen species are thought to contribute to pathogenesis of many cardiovascular diseases including hypertension, atherosclerosis, restenosis, heart failure, and diabetic vascular complications. Some of these reactive oxygen species also play an important role in vascular signaling. In this chapter, we describe various techniques that we have successfully employed to reliably measure superoxide and hydrogen peroxide. Because reactive oxygen species are capable of rapidly inactivating nitric oxide and because endothelial function characterized by nitric oxide bioavailability is an important indicator of vascular health, we have also included novel techniques capable of directly measuring nitric oxide radical from vascular cells and tissues. PMID- 18287683 TI - Assessment of PI-3 kinase and Akt in ischemic heart diseases in diabetes. AB - Diabetes mellitus is the most common disease in Westernized countries in large part because of the rising prevalence of obesity and physical inactivity. In addition, diabetes mellitus is an important risk factor for both heart failure and ischemic heart disease. As insulin resistance is known as an important pathophysiological feature in the cardiac diseases, understanding the mechanisms responsible for altered metabolism and insulin signaling in the diabetic heart may help identify novel targets in these conditions. Phosphatidylinositol (PI)-3 kinase (PI3K) and Akt are key signaling molecules in insulin and insulin-like growth factor-1 (IGF-1), which induce multiple biological effects in the heart such as cell survival and hypertrophy. Here, we have shown several fundamental techniques to study the role of PI3K and Akt in heart diseases. PMID- 18287684 TI - Two-dimensional gel electrophoresis in platelet proteomics research. AB - Proteomics technology allows a comprehensive and efficient analysis of the proteome and has become an indispensable tool in biomedical research. Since the late 80s, advances on mass spectrometry (MS) instrumentation and techniques have revolutionized the way proteins can be analyzed. Such analysis would only be possible with a proper sample preparation and separation ahead of the MS step. Different gel and nongel-based methods are available for protein separation. This chapter will focus on the use of two-dimensional gel electrophoresis (2-DE) in proteomics and its application to platelet research. 2-DE separates proteins according to their isoelectric point (pI) and size (molecular weight) and allows the detection of thousands of proteins at a time. Platelets are enucleated cells that play a critical function in the control of bleeding and wound healing. As platelets do not have a nucleus, proteomics offers a powerful alternative approach to provide data on protein expression in these cells, helping to address their biology. This chapter presents a protocol for an efficient sample preparation, protein separation by 2-DE, and protein digestion ahead of the MS analysis. The experimental approach, already successfully applied to the study of the platelet proteome, includes recommendations for an efficient platelet preparation for proteomics studies. PMID- 18287685 TI - Stem cell therapy in the heart and vasculature. AB - Stem cell therapy is a progressive approach to a pervasive clinical problem; cardiovascular disease is the number 1 killer in the USA and other developed countries, and aspects of it are amenable to stem cell therapy. Many types of stem cells have been used in treating the heart during myocardial infarction, and here, we describe our approach of direct myocardial injection of bone marrow derived mesenchymal stem cells into the infarct of rats. We will also briefly introduce the methods we have used to inject neonatal cardiomyocytes into the aorta as a first step in attempting to produce an external cardiac pump. Proper surgical technique and postoperative care are as important as adequate injection of the cells and will greatly improve the survival of the animal after surgery. By carefully following the methods presented in this chapter, the reader will be able to perform direct myocardial and vascular injection of stem cells into rats. PMID- 18287686 TI - The EMBL Nucleotide Sequence and Genome Reviews Databases. AB - Nucleotide and protein sequence databases are major resources for biological and medical research. This chapter introduces the European Molecular Biology Laboratory (EMBL) Nucleotide Sequence Database, a comprehensive primary data archive for nucleic acid sequences, and Genome Reviews, a secondary database that provides an up-to-date, standardized and comprehensively annotated view of the genomic sequence of selected organisms with completely deciphered genomes. Focusing on plant nucleotide sequences, we demonstrate how these data are accessed, how sequence similarity searches are performed and how we can obtain a wealth of additional information relating to genome sequences using Integr8. PMID- 18287687 TI - Using GenBank. AB - GenBank(R) is a comprehensive database of publicly available DNA sequences for more than 205,000 named organisms and for more than 60,000 within the embryophyta, obtained through submissions from individual laboratories and batch submissions from large-scale sequencing projects. Daily data exchange with the European Molecular Biology Laboratory (EMBL) in Europe and the DNA Data Bank of Japan ensures worldwide coverage. GenBank is accessible through the National Center for Biotechnology Information (NCBI) retrieval system, Entrez, which integrates data from the major DNA and protein sequence databases with taxonomy, genome, mapping, protein structure, and domain information and the biomedical journal literature through PubMed. BLAST provides sequence similarity searches of GenBank and other sequence databases. Complete bimonthly releases and daily updates of the GenBank database are available through FTP. GenBank usage scenarios ranging from local analyses of the data available through FTP to online analyses supported by the NCBI Web-based tools are discussed. To access GenBank and its related retrieval and analysis services, go to the NCBI Homepage at http://www.ncbi.nlm.nih.gov. PMID- 18287688 TI - A collection of plant-specific genomic data and resources at NCBI. AB - The National Center for Biotechnology Information (NCBI) provides a data-rich environment in support of genomic research by collecting the biological data for genomes, genes, gene expressions, gene variation, gene families, proteins, and protein domains and integrating the data with analytical, search, and retrieval resources through the NCBI Web site. Entrez, an integrated search and retrieval system, enables text searches across various diverse biological databases maintained at NCBI. Map Viewer, the genome browser developed at NCBI, displays aligned genetic, physical, and sequence maps for eukaryotic genomes including those of many plants. A specialized plant query page allows maps from all plant genomes available in the Map Viewer to be searched to produce a display of aligned maps from several species. Customized Plant Basic Local Alignment Search Tool (PlantBLAST) allows the user to perform sequence similarity searches in a special collection of mapped plant sequence data and to view the resulting alignments within a genomic context using Map Viewer. In addition, pre-computed sequence similarities, such as those for proteins offered by BLAST Link (BLink), enable fluid navigation from un-annotated to annotated sequences, quickening the pace of discovery. Plant Genome Central (PGC) is a Web portal that provides centralized access to all NCBI plant genome resources. Also, there are links to plant-specific Web resources external to NCBI such as organism-specific databases, genome-sequencing project Web pages, and homepages of genomic bioinformatics organizations. PMID- 18287689 TI - UniProtKB/Swiss-Prot. AB - The Swiss Institute of Bioinformatics (SIB), the European Bioinformatics Institute (EBI), and the Protein Information Resource (PIR) form the Universal Protein Resource (UniProt) consortium. Its main goal is to provide the scientific community with a central resource for protein sequences and functional information. The UniProt consortium maintains the UniProt KnowledgeBase (UniProtKB) and several supplementary databases including the UniProt Reference Clusters (UniRef) and the UniProt Archive (UniParc). (1) UniProtKB is a comprehensive protein sequence knowledgebase that consists of two sections: UniProtKB/Swiss-Prot, which contains manually annotated entries, and UniProtKB/TrEMBL, which contains computer-annotated entries. UniProtKB/Swiss-Prot entries contain information curated by biologists and provide users with cross links to about 100 external databases and with access to additional information or tools. (2) The UniRef databases (UniRef100, UniRef90, and UniRef50) define clusters of protein sequences that share 100, 90, or 50% identity. (3) The UniParc database stores and maps all publicly available protein sequence data, including obsolete data excluded from UniProtKB. The UniProt databases can be accessed online (http://www.uniprot.org/) or downloaded in several formats (ftp://ftp.uniprot.org/pub). New releases are published every 2 weeks. The purpose of this chapter is to present a guided tour of a UniProtKB/Swiss-Prot entry, paying particular attention to the specificities of plant protein annotation. We will also present some of the tools and databases that are linked to each entry. PMID- 18287690 TI - Plant database resources at The Institute for Genomic Research. AB - With the completion of the genome sequences of the model plants Arabidopsis and rice, and the continuing sequencing efforts of other economically important crop plants, an unprecedented amount of genome sequence data is now available for large-scale genomics studies and analyses, such as the identification and discovery of novel genes, comparative genomics, and functional genomics. Efficient utilization of these large data sets is critically dependent on the ease of access and organization of the data. The plant databases at The Institute for Genomic Research (TIGR) have been set up to maintain various data types including genomic sequence, annotation and analyses, expressed transcript assemblies and analyses, and gene expression profiles from microarray studies. We present here an overview of the TIGR database resources for plant genomics and describe methods to access the data. PMID- 18287691 TI - MIPS plant genome information resources. AB - The Munich Institute for Protein Sequences (MIPS) has been involved in maintaining plant genome databases since the Arabidopsis thaliana genome project. Genome databases and analysis resources have focused on individual genomes and aim to provide flexible and maintainable data sets for model plant genomes as a backbone against which experimental data, for example from high-throughput functional genomics, can be organized and evaluated. In addition, model genomes also form a scaffold for comparative genomics, and much can be learned from genome-wide evolutionary studies. PMID- 18287692 TI - HarvEST. PMID- 18287693 TI - The TAIR database. AB - The Arabidopsis Information Resource (TAIR) is a highly sophisticated, extensive, user friendly, Web-based resource for researchers working on the model plant Arabidopsis thaliana. The main gateway to this resource is through TAIR's homepage http://www.arabidopsis.org. It is a repository of large amounts of data including gene, mapping, protein, expression and community data in the form of a relational database. These data can be searched, downloaded and analysed using the tools provided. Here, the simple search (for retrieval of information), Seq Viewer (for the visualization of the five Arabidopsis chromosomes and associated annotations) and AraCyc (database of Arabidopsis biochemical pathways, with a graphical overview onto which large data sets, such as gene expression data, can be overlaid) tools are described with examples. PMID- 18287694 TI - AtEnsEMBL. AB - A comprehensive Arabidopsis genomic resource has been developed at the Nottingham Arabidopsis Stock Centre (NASC) to support the international plant community. This browser, termed AtEnsembl, provides a detailed and user-friendly interface for accessing a wide range of Arabidopsis-based genomic information and post genomic resources using the Ensembl browser. The resource aims to provide the broadest possible range of Ensembl features, including pointers to germplasm as well as representations of gene and protein information, links to Affymetrix gene expression data, and extensive data download capabilities. PMID- 18287695 TI - Accessing integrated Brassica genetic and genomic data using the BASC Server. AB - The BASC system provides tools for integrated mining and browsing of genetic, genomic and phenotypic data. The BASC demonstration server provides access to raw and analysed information for Brassica species and comparative information with Arabidopsis. We can use the tools within the Brassica BASC server to identify candidate genes for traits, conduct genome comparisons with Arabidopsis, identify syntenic regions and view gene expression profiles. The integration of BASC modules allows researchers to gain a comprehensive view of diverse Brassica genetic, genomic and phenotypic information. This chapter demonstrates the application of this resource, through the identification of candidate genes for an observed seed oil quality trait in Brassica napus. PMID- 18287696 TI - Leveraging model legume information to find candidate genes for soybean sudden death syndrome using the legume information system. AB - Comparative genomics is an emerging and powerful approach to achieve crop improvement. Using comparative genomics, information from model plant species can accelerate the discovery of genes responsible for disease and pest resistance, tolerance to plant stresses such as drought, and enhanced nutritional value including production of anti-oxidants and anti-cancer compounds. We demonstrate here how to use the Legume Information System for a comparative genomics study, leveraging genomic information from Medicago truncatula (barrel medic), the model legume, to find candidate genes involved with sudden death syndrome (SDS) in Glycine max (soybean). Specifically, genetic maps, physical maps, and annotated tentative consensus and expressed sequence tag (EST) sequences from G. max and M. truncatula can be compared. In addition, the recently published M. truncatula genomic sequences can be used to identify M. truncatula candidate genes in a genomic region syntenic to a quantitative trait loci region for SDS in soybean. Genomic sequences of candidate genes from M. truncatula can then be used to identify ESTs with sequence similarities from soybean for primer design and cloning of potential soybean disease causing alleles. PMID- 18287697 TI - Legume Resources: MtDB and Medicago.Org. AB - To identify the genes and gene functions that underlie key aspects of legume biology, researchers have selected the cool season legume Medicago truncatula as a model system for legume research. The mission of the M. truncatula Consortium is to promote unrestricted sharing of data and information that are provided by Medicago research groups worldwide. Through integration of a variety of data and tools, the medicago.org site intends to facilitate progress in the fields of structural, comparative, and functional genomics. To this goal, and as a consortium partner, the Center for Computational Genomics and Bioinformatics (CCGB) at the University of Minnesota has developed MtDB2.0, the M. truncatula database version 2.0. The MtDB2.0 database is the first step toward the global integration of M. truncatula genomic, genetic, and biological information. MtDB2.0 is a relational database that integrates M. truncatula transcriptome data and provides a wide range of user-defined data mining options. The database is interrogated through a series of interfaces, with 58 options grouped into two filters. Sequence identifiers from all public M. truncatula sites [e.g., IDs from GenBank, CCGB, The Institute for Genomic Research (TIGR), National Center for Genome Resources (NCGR), and I'Institut National de la Recherche Agronomique (INRA)] are fully cross-referenced to facilitate comparisons between different sites, and hypertext links to the appropriate database records are provided for all queries' results. MtDB's goal is to provide researchers with the means to quickly and independently identify sequences that match specific research interests based on user-defined criteria. MtDB2.0 offers unrestricted access to advanced and powerful querying tools unmatched by any other public databases. Structurad Query Language (SQL)-encoded queries with a Java-based Web user interface, incorporate different filtering that allow sophisticated data mining of the expressed sequence tag sequencing project results, including the CCGB M. truncatula Unigene set generated with the Phrap assembler. The underlying database and query software have been designed for ease of updates and portability to other model organisms. Public access to the database is at http://www.medicago.org/MtDB. PMID- 18287698 TI - BGI-RIS V2. AB - Rice serves as both a staple for over half of the world's population and a model organism for plants of the grass family. Beijing Genomics Institute (BGI) has long been engaged in rice genomic research: sequencing, assembly, information analysis and integration. Such intensive research results in public releases and biological applications. In order to facilitate obtaining and operating on the rice genomic data, as well as to provide a genomic groundwork for comparative, functional or evolutionary research on important cereal crops, BGI has established and updated the Rice Information System (BGI-RIS V2), an integrated information resource and comparative analysis workbench for rice genomes. BGI-RIS V2 offers not only genomic sequences, which combine the genomic data of Oryza sativa L. ssp. indica (by BGI) with Oryza sativa L. ssp. japonica, but also most detailed annotation data, including genetic markers, Bacterial Artificial Chromosome (BAC) end sequences, gene contents, cDNAs, oligos, tiling arrays, repetitive elements, and genomic polymorphisms. As a basic platform, BGI-RIS V2 also offers graphical interfaces and a series of tools and services for gene finding, genomic alignment and genomic assembly. This database is available through the web server (http://rise.genomics.org.cn or http://rice.genomics.org.cn) and the File Transfer Protocol (FTP) server (ftp://ftp.genomics.org.cn/pub/database/rice). PMID- 18287699 TI - GrainGenes. AB - The GrainGenes website hosts a wealth of information for researchers working on Triticeae species, oat and their wild relatives. The website hosts a database encompassing information such as genetic maps, genes, alleles, genetic markers, phenotypic data, quantitative trait loci studies, experimental protocols and publications. The database can be queried by text searches, browsing, Boolean queries, MySQL commands, or by using pre-made queries created by the curators. GrainGenes is not solely a database, but serves as an informative site for researchers and a means to communicate project aims, outcomes and a forum for discussion. This chapter describes the type of information available on the site and database, and the options available to access this data. PMID- 18287700 TI - Gramene. AB - Grasses are one of the largest agricultural crops, providing food, industrial materials and renewable energy sources. Due to their large genome size and the number of the species in the taxa, many of the genomes are not targeted for complete sequencing. Gramene seeks to provide basic researchers, industry and educators with a resource that can be used as a tool for knowledge discovery across grass species. This chapter briefly outlines system requirements for end users and database hosting, outlines data types and basic navigation within Gramene and provides an example of how a maize researcher would use Gramene to leverage rice genome organization and phenotypic information to support targeted experimental research in maize. PMID- 18287701 TI - MaizeGDB. AB - MaizeGDB is the community database for biological information about the crop plant Zea mays. Genetic, genomic, sequence, gene product, functional characterization, literature reference, and person/organization contact information are among the data types stored at MaizeGDB. At the project's website (http://www.maizegdb.org) are standardized custom interfaces enabling researchers to browse data and to seek out specific information matching explicit search criteria. In addition, pre-compiled reports are made available for particular types of data, and bulletin boards are provided to facilitate communication and coordination among members of the community of maize geneticists. PMID- 18287702 TI - BarleyBase/PLEXdb. AB - BarleyBase (http://barleybase.org/) and its successor, PLEXdb (http://plexdb.org/), are public resources for large-scale gene expression analysis for plants and plant pathogens. BarleyBase/PLEXdb provides a unified web interface to support the functional interpretation of highly parallel microarray experiments integrated with traditional structural genomics and phenotypic data. Users can perform hypothesis building queries from multiple interlinked resources, e.g., a particular gene, a protein class, EST entries, and physical or genetic map position-all coupled to highly parallel gene expression, for a variety of crop and model plant species, from a large array of experimental or field conditions. Array data are interlinked to analytical and biological functions (e.g., Gene and Plant Ontologies, BLAST, spliced alignment, multiple alignment, regulatory motif identification, and expression analysis), allowing members of the community to access and analyze comparative expression experiments in conjunction with their own data. PMID- 18287703 TI - Reaping the Benefits of SAGE. AB - Serial analysis of gene expression (SAGE) is a powerful technique which yields a digital measure of gene expression through the sequencing of libraries of specific mRNA-derived fragments, namely SAGE tags. This chapter introduces the methods and software tools that are available for researchers to analyze gene expression through SAGE analysis. A detailed examination of SAGE analysis in Arabidopsis thaliana using the publicly available analysis tool, SaskSAGE, is provided. The use of this software allows the user to maximize the information gained from SAGE experiments in a model system with a fully sequenced genome. PMID- 18287704 TI - Methods for analysis of gene expression in plants using MPSS. AB - Massively parallel signature sequencing (MPSS) is a technology capable of sequencing simultaneously almost all the DNA molecules in a sample. This technology is well suited for deep profiling of mRNA and small RNA by the sequencing of cDNA tags. A series of mRNA MPSS databases has been created from various libraries derived from four different species (Arabidopsis, rice, grape, and Magnaporthe grisea, the rice blast fungus). Our mRNA MPSS databases measure the absolute expression level of most genes in the sample and provide information about potentially novel transcripts (antisense transcripts, alternative splice isoforms, and regulatory intergenic transcripts). In addition to these data, we have recently built an extensive database from MPSS-derived Arabidopsis small RNA samples. All the databases are accessible thorough our Web interface (http://mpss.udel.edu), and the individual pages are equipped with various graphical and analytical tools. Here, we focus on a subset of these tools (e.g., Gene Analysis [GA], Chromosome Viewer [CV], and Library Analysis [LIBAN]) and describe how the users can analyze and interpret our MPSS expression data. PMID- 18287705 TI - Metabolomics data analysis, visualization, and integration. AB - Metabolomics is the large-scale analysis of metabolites and as such requires bioinformatics tools for data analysis, visualization, and integration. This chapter describes the basic composition of chromatographically coupled mass spectrometry (MS) data sets used in metabolomics and describes in detail the steps necessary for extracting large-scale qualitative and quantitative information. This process involves noise filtering, peak picking and deconvolution, peak identification, peak alignment, and the creation of a final data matrix for statistical processing. Multivariate tools for comparative analysis are presented and illustrated using data for Medicago truncatula. Additional tools for visualizing and integrating metabolomics data within a biological context are discussed. Two tables are provided listing current metabolomics data processing and visualization software. Because metabolomics is rapidly maturing, a final section is presented concerning the need for data standardization and current efforts. PMID- 18287706 TI - KEGG bioinformatics resource for plant genomics research. AB - Kyoto Encyclopedia of Genes and Genomes (KEGG) is a bioinformatics resource for understanding biological function from a genomic perspective. It is a multispecies, integrated resource consisting of genomic, chemical, and network information, with cross-references to numerous outside databases and containing a complete set of building blocks (genes and molecules) and wiring diagrams (biological pathways) to represent cellular functions. KEGG consists of a suite of databases: PATHWAY, GENES/Sequence Similarity Database (SSDB), Biomolecular Relations in Information Transmission and Expression (BRITE), and LIGAND, which is a composite database of COMPOUND, DRUG, GLYCAN, REACTION, REPAIR, and ENZYME. Two new databases have been recently added to KEGG: DGENES (for draft genomes) and EGENES (for expressed-sequence tag [EST] data). EGENES is a knowledge base system for efficient analysis of organism-specific ESTs, including publicly available plant ESTs. EGENES links the genomic information with higher order functional information in a single database. The genomic information stored in EGENES is a collection of EST contigs, produced by assembling the public ESTs. In this chapter, we will introduce KEGG and discuss its importance for the plant research community by focusing on EGENES. Because all the resources in KEGG follow the same architecture and design, an appraisal of EGENES should give readers an idea of the available information stored in KEGG and how to use them efficiently. PMID- 18287707 TI - International crop information system for germplasm data management. AB - Passport and phenotypic data on germplasm and breeding lines are available from worldwide sources in various electronic formats. These data can be collated into a single database format to enable strategic interrogation to make the best use of data for effective germplasm use and enhancement. The International Crop Information System (http://www.icis.cgiar.org) is an open-source project under development by a global community of crop researchers and includes applications designed to achieve the storage and interrogation of pedigree and phenotypic data. PMID- 18287708 TI - Automated discovery of single nucleotide polymorphism and simple sequence repeat molecular genetic markers. AB - Molecular genetic markers represent one of the most powerful tools for the analysis of genomes. Molecular marker technology has developed rapidly over the last decade, and two forms of sequence-based markers, simple sequence repeats (SSRs), also known as microsatellites, and single nucleotide polymorphisms (SNPs), now predominate applications in modern genetic analysis. The availability of large sequence data sets permits mining for SSRs and SNPs, which may then be applied to genetic trait mapping and marker-assisted selection. Here, we describe Web-based automated methods for the discovery of these SSRs and SNPs from sequence data. SSRPrimer enables the real-time discovery of SSRs within submitted DNA sequences, with the concomitant design of PCR primers for SSR amplification. Alternatively, users may browse the SSR Taxonomy Tree to identify predetermined SSR amplification primers for any species represented within the GenBank database. SNPServer uses a redundancy-based approach to identify SNPs within DNA sequence data. Following submission of a sequence of interest, SNPServer uses BLAST to identify similar sequences, CAP3 to cluster and assemble these sequences, and then the SNP discovery software autoSNP to detect SNPs and insertion/deletion (indel) polymorphisms. PMID- 18287709 TI - Methods for gene ontology annotation. AB - The Gene Ontology (GO) is an established dynamic and structured vocabulary that has been successfully used in gene and protein annotation. Designed by biologists to improve data integration, GO attempts to replace the multiple nomenclatures used by specialised and large biological knowledgebases. This chapter describes the methods used by groups to create new GO annotations and how users can apply publicly available GO annotations to enhance their datasets. PMID- 18287710 TI - Gene Structure Annotation at PlantGDB. AB - The accurate identification of exons and introns that comprise a complete plant gene structure can be a time-consuming and challenging task. Novel Web-based tools facilitate the process by providing a convenient interface to current transcript evidence, and portals to relevant bioinformatics software. With a few keystrokes, the user can explore alternative transcript assemblies and, for example, select for annotation those that are clearly supported by transcript evidence and similarity to known genes. The implementation of the tool at the PlantGDB resource also allows immediate communication of the novel annotations to the community through Web display. PMID- 18287711 TI - An Introduction to BioPerl. AB - The BioPerl toolkit provides a library of hundreds of routines for processing sequence, annotation, alignment, and sequence analysis reports. It often serves as a bridge between different computational biology applications assisting the user to construct analysis pipelines. This chapter illustrates how BioPerl facilitates tasks such as writing scripts summarizing information from BLAST reports or extracting key annotation details from a GenBank sequence record. PMID- 18287743 TI - Isolation and characterization of lipopolysaccharides. AB - Lipopolysaccharide (LPS) is the signature glycolipid isolated from almost all Gram-negative bacteria. LPSs are well known for their ability to elicit the release of cytokines from eukaryotic cells including macrophages, neutrophils, and epithelial cells. LPS can be isolated free of contaminating nucleic acids and proteins by various techniques. In this review, we outline approaches for the isolation and preparation of LPSs for structural studies as well as preparation of very highly purified material for biological studies. Methods are also provided for the analysis of the purity and the structural composition of the LPSs. Finally, three methods for the isolation of lipid A are described. PMID- 18287744 TI - Proteomic analysis of proteins secreted by Streptococcus pyogenes. AB - Streptococcus pyogenes secretes various proteins to the extracellular environment. During infection, these proteins interact with human macromolecules and contribute to pathogenesis. We describe a proteomic approach routinely used in our laboratory to characterize culture supernatant proteins using small-format two-dimensional gel electrophoresis. Proteins are collected after overnight growth of the bacteria in broth media. Compounds that inhibit isoelectric focusing, such as salts, are removed by enzymatic treatment and precipitation with trichloroacetic acid and acetone. Following resuspension in denaturing solution, the proteins are separated by isoelectric focusing using a 7-cm immobilized strip with a pH gradient of 4-7. Subsequently, proteins are further separated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) and stained with SYPRO Ruby. The small-gel format requires less time, reagents, and smaller culture volumes compared with large-format approaches, while still resolving and detecting a large proportion of the exoprotein fraction. PMID- 18287745 TI - Proteomic analysis to investigate regulatory networks in Staphylococcus aureus. AB - The analysis of the expression of virulence genes and the elucidation of metabolic and regulatory pathways of Staphylococcus aureus provide us with important information about the interaction between the pathogen and its host, mechanisms by which this organism causes diseases, and the resistance to antibiotics. In order to investigate regulatory networks of S. aureus, we analyze the cytoplasmic and extracellular proteome by using two-dimensional (2D) gel analyses combined with matrix-assisted laser ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Gel-based proteomics is an extremely valuable tool in microbial physiology that can, in combination with various visualization and quantitation software packages, very rapidly provide comparative and quantitative data for multi-sample comparison. PMID- 18287746 TI - Microarray comparative genomic hybridization for the analysis of bacterial population genetics and evolution. AB - Comparative genomic hybridization analyses have contributed greatly to our understanding of bacterial evolution, population genetics, and pathogenesis. Here, we describe a robust protocol for microarray-based comparison of genome content, which could be applied to any bacterial species of interest. PMID- 18287747 TI - Detection and inhibition of bacterial cell-cell communication. AB - Bacteria communicate with other members of their community through the secretion and perception of small chemical cues or signals. The recognition of a signal normally leads to the expression of a large suite of genes, which in some bacteria are involved in the regulation of virulence factors, and as a result, these signaling compounds are key regulatory factors in many disease processes. Thus, it is of interest when studying pathogens to understand the mechanisms used to control the expression of virulence genes so that strategies might be devised for the control of those pathogens. Clearly, the ability to interfere with this process of signaling represents a novel approach for the treatment of bacterial infections. There is a broad range of compounds that bacteria can use for signaling purposes, including fatty acids, peptides, N-acylated homoserine lactones, and the signals collectively called autoinducer 2 (AI-2). This chapter will focus on the latter two signaling systems as they are present in a range of medically relevant bacteria, and here we describe assays for determining whether an organism produces a particular signal and assays that can be used to identify inhibitors of the signaling cascade. Lastly, the signal detection and inhibition assays will be directly linked to the expression of virulence factors of specific pathogens. PMID- 18287748 TI - A system for site-specific genetic manipulation of the relapsing fever spirochete Borrelia hermsii. AB - The lack of a system for genetic manipulation has hindered studies on the molecular pathogenesis of relapsing fever Borrelia. The focus of this chapter is to describe selectable markers, manipulation strategies, and methods to electro transform and clone wild-type infectious Borrelia hermsii. Preliminary studies suggest that the variable tick protein (Vtp) of B. hermsii is involved in tick-to mammal transmission. To address this hypothesis, we have developed a system for genetic manipulation and have constructed clones of a Vtp mutant and an isogenic reconstituted strain. The methods described here are applicable for the inactivation of other loci in B. hermsii and should be adaptable for other species of relapsing fever spirochetes. PMID- 18287749 TI - Transposon mutagenesis of the lyme disease agent Borrelia burgdorferi. AB - Borrelia burgdorferi, the causative agent of Lyme disease, is an obligate parasite that cycles between vertebrate hosts and tick vectors. Attempts to understand the genetic factors that allow B. burgdorferi to sense, adapt to, and survive in different environments have been limited by a relatively low transformation rate. Here, we describe a mariner-based transposon system that achieves saturating levels of random mutagenesis in B. burgdorferi. In comparison with allelic exchange, which targets a single locus, transposon mutagenesis can create libraries of mutants encompassing disruptions of all genes. Suitably designed screens or selections of such a library permit the recovery of mutants exhibiting a desired phenotype. The system described here allows rapid identification of the genetic locus responsible for the mutant phenotype. With appropriate modifications, this mariner-based transposon can be adapted to other spirochetes and bacteria with inefficient genetic transformation methods. PMID- 18287750 TI - The biofilm exopolysaccharide polysaccharide intercellular adhesin--a molecular and biochemical approach. AB - Exopolysaccharides play a crucial role in the formation of biofilms and biofilm resistance to antimicrobials and innate host defense. Here we describe methods to analyze and quantify polysaccharide intercellular adhesin (PIA), a biofilm exopolysaccharide made of N-acetylglucosamine that is found in staphylococci and many other bacterial biofilm-forming pathogens. PMID- 18287751 TI - Analysis of Staphylococcus aureus gene expression during PMN phagocytosis. AB - Staphylococcus aureus is a leading cause of human infections worldwide and causes a variety of diseases ranging in severity from mild to life-threatening. The ability of S. aureus to cause disease is based in part on its ability to subvert the innate immune system. Advances in genome-wide analysis of host-pathogen interactions have provided the necessary tools to investigate molecular factors that directly contribute to S. aureus pathogenesis. This chapter describes methods to analyze gene expression in S. aureus during interaction with human neutrophils. PMID- 18287752 TI - Examining the vector-host-pathogen interface with quantitative molecular tools. AB - We developed PCR assays to detect and quantitate Yersinia pestis, the bacterial agent of plague, in flea vector and mammalian host tissues. Bacterial numbers in fleas, fleabite sites, and infected lymph nodes were determined using real-time PCR with primers and probes for a gene target on a multi-copy plasmid specific to Y. pestis. Tissue-matched standard curves used to determine absolute bacterial numbers in unknown samples were linear over at least five orders of magnitude. The methods were applied to studies of transmission of Y. pestis by the rat flea Xenopsylla cheopis, but should be generally useful to investigate the transmission dynamics of any arthropod-borne disease. PMID- 18287753 TI - Intracellular localization of Brucella abortus and Francisella tularensis in primary murine macrophages. AB - Intracellular bacterial pathogens have evolved sophisticated strategies to survive and proliferate within cells of their hosts. Studying their intracellular life cycle is key to understanding virulence and requires methodologies that can identify the compartments in which they localize and characterize the replicative niche they generate. Here, we describe immunofluorescence-based microscopy techniques applied to the intracellular pathogens Brucella abortus and Francisella tularensis during their respective intracellular cycles inside murine bone marrow-derived macrophages. Standard immunofluorescence techniques are used to define the intracellular localization of the pathogens based on their co localization with specifically labeled macrophage organelles. In addition, we describe an assay to assess the integrity of Francisella-containing phagosomes and bacterial release into the macrophage cytoplasm, which is a hallmark of Francisella intracellular pathogenesis. PMID- 18287754 TI - Rate and extent of Helicobacter pylori phagocytosis. AB - Helicobacter pylori is a Gram-negative bacterium that colonizes the gastric epithelium and plays a causative role in the development of peptic ulcers and gastric cancer. Phagocytosis is an element of innate defense used by macrophages and neutrophils to engulf microorganisms. We and others have shown that strains of H. pylori that contain the cag pathogenicity island actively retard their entry into phagocytes. Consequently, there is a lag of several minutes between bacterial binding and the onset of engulfment, and relative to other particles and microbes, the rate of internalization is slow. Herein, we describe in detail the use of synchronized phagocytosis and indirect immunofluorescence microscopy to quantify the rate and extent of H. pylori phagocytosis. This method is appropriate for primary phagocytes as well as transformed cell lines. More importantly, the effects of opsonins, virulence factors, and other agents on infection can be measured independent of bacterial viability or intracellular locale. PMID- 18287755 TI - Culture, isolation, and labeling of Anaplasma phagocytophilum for subsequent infection of human neutrophils. AB - Anaplasma phagocytophilum is the etiologic agent of granulocytic anaplasmosis, a tick-borne, zoonotic, emerging infectious disease. A. phagocytophilum is an obligate intracellular pathogen that primarily resides within membrane-bound, cytoplasmic vacuoles of host neutrophils. Closely related to Ehrlichial and Rickettsial organisms, A. phagocytophilum is a small, fragile, Gram-negative bacterium that presents unique challenges for culture, isolation, enumeration, and labeling. This chapter delineates pathogen-specific considerations for culture and labeling of this organism for subsequent use in assays to examine mechanisms of host cell-pathogen interactions. PMID- 18287756 TI - Ultrastructural analysis of bacteria-host cell interactions. AB - Electron microscopy of bacterial pathogens and interactions between bacteria and host cells and tissues provides valuable insights into structural and molecular properties and processes involved in pathogenesis. Applications for electron microscopy in bacterial pathogenesis range from discovering etiologic agents and following chronological events during infections by conventional examination of clinical samples to assessing molecular host-cell responses to infection and in situ interactions between receptors and ligands using specific immune-labeling techniques. This chapter focuses on techniques for preparing samples of bacteria and host cells for conventional transmission (TEM) and scanning electron microscopy (SEM) and use of luminescent nanocrystals or "quantum dots" as specific probes for correlative light and electron microscopy. Conventional TEM and SEM are well established tools for high resolution examination of structural effects and chronological events associated with bacterial infections. The recent development of quantum dots as physiological and immunological probes in biology has provided a powerful technique for bridging fluorescent analyses of fixed and live material with preparation and examination by TEM and SEM. PMID- 18287757 TI - Infection of human monocyte-derived macrophages with Coxiella burnetii. AB - Coxiella burnetii, the agent of Q fever, is an obligate intracellular bacterium that has a tropism for cells of the mononuclear phagocyte system. Following internalization, C. burnetii remains in a phagosome that ultimately matures into a vacuole with lysosomal characteristics that supports pathogen replication. Most in vitro investigations of Coxiella - macrophage interactions have employed continuous cell lines. Although these studies have been informative, genetic alterations of immortalized cells may result in attenuated biological responses to infection relative to primary cells. Consequently, primary macrophages are preferred as in vitro model systems. Here, we describe procedures for propagation and isolation of C. burnetii from cell culture and the use of these preparations to infect primary macrophages derived from human peripheral blood monocytes. Both virulent phase I and avirulent phase II C. burnetii productively infect human monocyte-derived macrophages (MDMs) and replicate with approximately the same kinetics, thereby providing a more physiologically relevant in vitro model system to study the infectious process of this pathogen. PMID- 18287758 TI - Infection of epithelial cells with Salmonella enterica. AB - Salmonella enterica serovars cause a variety of diseases ranging from self limiting gastroenteritis to severe systemic infections. Virulence of these facultative intracellular pathogens is dependent on their ability to invade and replicate within non-phagocytic cells, and cultured epithelial cell systems have been used extensively to dissect the molecular mechanisms involved. For efficient invasion in vitro, the bacterial cell growth conditions as critical since the invasion associated type III secretion system (T3SS1) must be expressed and functional. The ability of Salmonella to invade, and replicate within, epithelial cells can be easily assessed using a gentamicin protection assay or immunofluorescence microscopy. Here, the protocols used in our laboratory are described in detail. PMID- 18287759 TI - Determining the cellular targets of reactive oxygen species in Borrelia burgdorferi. AB - The response of Borrelia burgdorferi to the challenge of reactive oxygen species (ROS) is a direct result of its limited biosynthetic capabilities and lack of biologically significant levels of intracellular Fe. In other bacteria, the major target for oxidative damage is DNA as a consequence of the reaction of "free" intracellular with ROS through the Fenton reaction. Therefore, cellular defenses in these bacteria are focused on protecting this essential cellular component. This does not seem to be the case for B. burgdorferi. In this chapter, we describe methods that were used to analyze the potential targets for ROS in B. burgdorferi. Surprisingly, membrane lipids (e.g., linoleic and linolenic acids) derived from host are the major target of ROS in the Lyme disease spirochete. PMID- 18287760 TI - Bioluminescent imaging of bacterial biofilm infections in vivo. AB - Whole body biophotonic imaging (BPI) is a technique that has contributed significantly to the way researchers study bacterial pathogens and develop pre clinical treatments to combat their ensuing infections in vivo. Not only does this approach allow disease profiles and drug efficacy studies to be conducted non-destructively in live animals over the entire course of the disease, but in many cases, it enables investigators to observe disease profiles that could otherwise easily be missed using conventional methodologies. The principles of this technique are that bacterial pathogens engineered to express bioluminescence (visible light) can be readily monitored from outside of the living animal using specialized low-light imaging equipment, enabling their movement, expansion and treatment to be seen completely non-invasively. Moreover, because the same group of animals can be imaged at each time-point throughout the study, the overall number of animals used is dramatically reduced, saving lives, time, and money. Also, as each animal acts as its own control over time, the issues associated with animal-to-animal variation are circumvented, thus improving the quality of the biostatistical data generated. The ability to monitor infections in vivo in a longitudinal fashion is especially appealing to assess chronic infections such as those involving implanted devices. Typically, bacteria grow as biofilms on these foreign bodies and are reputably difficult to monitor with conventional methods. Because of the non-destructive and non-invasive nature of BPI, the procedure can be performed repeatedly in the same animal, allowing the biofilm to be studied in situ without detachment or disturbance. This ability not only allows unique patterns of disease relapse to be seen following termination of antibiotic therapy but also in vivo resistance development during prolonged treatment, both of which are common occurrences with device-related infections. This chapter describes the bioluminescent engineering of both Gram-positive and Gram-negative bacteria and overviews their use in device-associated infections in several anatomical sites in a variety of animal models. PMID- 18287761 TI - The Cotton Rat as a Model for Staphylococcus aureus nasal colonization in humans: cotton rat S. aureus nasal colonization model. AB - Staphylococcus aureus nasal colonization is a well-known risk factor for development of S.aureus infections in humans, but despite this established association, we are only beginning to understand the factors, both host and pathogen, that play a role in the colonization of the nares by S. aureus. The cotton rat is a model for many human respiratory pathogens and has proved its utility as a robust model for S. aureus nasal colonization. In this animal model, S. aureus is instilled in the nostrils of adult cotton rats, the bacteria rapidly colonize, and 7 days later S. aureus nasal colonization is enumerated by surgical removal of the nose and recovery of the colonizing S. aureus. This model is an excellent animal model to allow for the evaluation of the efficacy of various therapies, including semi-solid formulations, for determination of their ability to eradicate S. aureus nasal colonization. Further, the cotton rat model allows for assessment of the ability of defined genetic mutants of S. aureus to colonize mucosal surfaces. Finally, this model has demonstrated its utility for the assessment of various antigens as vaccine candidates to protect against S. aureus nasal colonization. This chapter will discuss in detail the method to establish nasal colonization, treatment and eradication of colonization, and recovery of the colonizing bacteria from the nose. PMID- 18287762 TI - A non-human primate model of acute group a Streptococcus pharyngitis. AB - This chapter describes methods for using non-human primates as a model of group A streptococcal (GAS) pharyngitis. This model has been used successfully to study host-pathogen interactions occurring during pharyngeal GAS infections. The protocol as described will compare two different GAS strains for their ability to cause clinical symptoms of pharyngitis. PMID- 18287763 TI - Target-based antimicrobial drug discovery. AB - The continued increase in antibiotic resistance among bacterial pathogens, coupled with a decrease in infectious disease research among pharmaceutical companies, has escalated the need for novel and effective antibacterial chemotherapies. While current agents have emerged almost exclusively from whole cell screening of natural products and small molecules that cause microbial death, recent advances in target identification and assay development have resulted in a flood of target-driven drug discovery methods. Whether genome-based methodologies will yield new classes of agents that conventional methods have been unable to is yet to be seen. At the end of the day, perhaps a synergy between old and new approaches will harvest the next generation of antibacterial treatments. PMID- 18287764 TI - Sequence analysis of the variable number tandem repeat in Staphylococcus aureus protein A gene: spa typing. AB - The analyses of numerous prokaryotic and eukaryotic genomes have revealed the presence of variable number tandem repeats (VNTRs). VNTR analysis is currently widely used to sub-speciate many bacterial, fungal, and viral pathogens and has facilitated a number of molecular epidemiology studies. In this chapter, we focus on spa typing which is based on sequence analysis of VNTRs in the polymorphic X region of the Staphylococcus aureus protein A gene Staphylococcus aureus. As the specific methods for spa typing, detailed in this chapter, are well-established and routine procedures (e.g., DNA isolation, PCR and DNA sequencing) for most molecular biology laboratories, we highlight the analytic methods used to interpret the genotyping data generated by sequence analysis and their potential applications in local and global epidemiologic investigations. PMID- 18287765 TI - Overview and introduction to clinical proteomics. AB - As the field of clinical proteomics progresses, discovery of disease biomarkers becomes paramount. However, the immediate challenges are to establish standard operating procedures for both clinical specimen handling and reduction of sample complexity and to increase the ability to detect proteins and peptides present in low amounts. The traditional concept of a disease biomarker is shifting toward a new paradigm, namely, that an ensemble of proteins or peptides would be more efficient than a single protein/peptide in the diagnosis of disease. Because clinical proteomics usually requires easy access to well-defined fresh clinical specimens (including morphologically consistent tissue and properly pretreated body fluids of sufficient quantity), biorepository systems need to be established. Here, we address these questions and emphasize the necessity of developing various microdissection techniques for tissue specimens, multidimensional fractionation for body fluids, and other related techniques (including bioinformatics), tools which could become integral parts of clinical proteomics for disease biomarker discovery. PMID- 18287766 TI - Specimen collection and handling: standardization of blood sample collection. AB - Preanalytical variables can alter the analysis of blood-derived samples. Prior to the analysis of a blood sample, multiple steps are necessary to generate the desired specimen. The choice of blood specimens, its collection, handling, processing, and storage are important aspects since these characteristics can have a tremendous impact on the results of the analysis. The awareness of clinical practices in medical laboratories and the current knowledge allow for identification of specific variables that affect the results of a proteomic study. The knowledge of preanalytical variables is a prerequisite to understand and control their impact. PMID- 18287767 TI - Tissue sample collection for proteomics analysis. AB - Successful collection of tissue samples for molecular analysis requires critical considerations. We describe here our procedure for tissue specimen collection for proteomic purposes with emphasis on the most important steps, including timing issues and the procedures for immediate freezing, storage, and microdissection of the cells of interest or "tissue targets" and the lysates for protein isolation for SELDI, MALDI, and 2DGE applications. The pathologist is at the cornerstone of this process and is an invaluable collaborator. In most institutions, pathologists are responsible for "tissue custody," and they closely supervise the tissue bank. In addition, they are optimally trained in histopathology in order to they assist investigators to correlate tissue morphology with molecular findings. In recent years, the advent of the laser capture microscope, a tool ideally designed for pathologists, has tremendously facilitated the efficiency of collecting tissue targets for molecular analysis. PMID- 18287768 TI - Protein profiling of human plasma samples by two-dimensional electrophoresis. AB - Human plasma is regarded the most complex and well-known clinical specimen that can be easily obtained; alterations in the levels of plasma proteins or their corresponding enzyme activities may reflect either a healthy or a diseased state. Given that there is no defined genomic information as to the intact protein components in plasma, protein profiling could be the first step toward its molecular characterization. Several problems exist in the analysis of plasma proteins, however. For example, the widest dynamic range of protein concentrations, the presence of high-abundance proteins, and post-translational modifications need to be considered before proteomic studies are undertaken. In particular, efficient depletion or pre-fractionation of high-abundance proteins is crucial for the identification of low-abundance proteins that may contain potential biomarkers. After the removal of high-abundance proteins, protein profiling can be initiated using two-dimensional electrophoresis (2DE), which has been widely used for displaying the differential proteome under specific physiological conditions. Here, we describe a typical 2DE procedure for plasma proteome under either a healthy or a diseased state (e.g., liver cancer) in which pre-fractionation and depletion are integral steps in the search for disease biomarkers. PMID- 18287769 TI - Analysis of laser capture microdissected cells by 2-dimensional gel electrophoresis. AB - Laser capture microdissection (LCM) is a powerful tool for procuring near-pure populations of targeted cell types from specific microscopic regions of tissue sections, by overcoming problems due to tissue heterogeneity and minimizing intermixture and contamination by other cell types. The combination of LCM with various proteomic technologies has enabled high-throughput molecular analysis of human tumors, and provided critical tools in the search for novel disease markers and therapeutic targets. As an example, we describe the application of LCM in dissecting the tumor cells in breast cancer for macromolecular extraction and subsequent protein separation by 2-dimensional gel electrophoresis (2-D GE). The protocols and the key issues involved in preparing ethanol-fixed paraffin embedded tissue blocks and microscopic sections, microdissecting the cells of interest using the PixCell II LCM system, extracting and separating the cellular proteins by 2-D GE, and preparing selective proteins for peptide mass analysis by mass spectrometry, are discussed. The aim is to provide a practical guide in performing high-throughput microdissection of target cells and gel-based proteomics, which can be adapted to research in cancer formation and growth. PMID- 18287770 TI - Optimizing the difference gel electrophoresis (DIGE) technology. AB - Difference gel electrophoresis (DIGE) technology has been used to provide a powerful quantitative component to proteomics experiments involving 2D gel electrophoresis. DIGE combines spectrally resolvable fluorescent dyes (Cy2, Cy3, and Cy5) with sample multiplexing for low technical variation, and uses an internal standard methodology to analyze replicate samples from multiple experimental conditions with unsurpassed statistical confidence for 2D gel-based differential display proteomics. DIGE experiments can facilely accommodate sufficient independent (biological) replicate samples to control for the large interpersonal variation expected from clinical samples. The use of multivariate statistical analyses can then be used to assess the global variation in a complex set of independent samples, filtering out the noise from technical variation and normal biological variation thereby focusing on the underlying variation that can describe different disease states. This chapter focuses on the design and implementation of the DIGE methodology employing the use of a pooled-sample internal standard in conjunction with the minimal CyDye chemistry. Notes are also provided for the use of the alternative saturation labeling chemistry. PMID- 18287771 TI - MALDI/SELDI protein profiling of serum for the identification of cancer biomarkers. AB - The ability to visualize the full depth of the serum proteome in a high throughput manner is a major goal of clinical proteomics. Methodologies, which combine higher throughput with the ability to observe differential protein expression levels, have been applied to this goal. An example of such a system is the coupling of robotic sample processing to matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF-MS). Within this paradigm is a modification of MALDI-TOF termed surface-enhanced laser desorption/ionization-TOF (SELDI-TOF). Both conventional MALDI and SELDI have been used to generate protein expression profiles reflective of potential peptide changes in serum. This information can be used to identify proteins, which may enable new diagnostic and therapeutic strategies. PMID- 18287772 TI - Urine sample preparation and protein profiling by two-dimensional electrophoresis and matrix-assisted laser desorption ionization time of flight mass spectroscopy. AB - Urine represents the most easily attainable and consequently one of the most common samples in clinical analysis and diagnostics. However, urine is also considered one of the most difficult proteomic samples to work with due to its highly variable contents, as well as the presence of various proteins in low abundance or modified forms. In this chapter, we describe simple protocols and troubleshooting tips for urinary protein preparation and profiling by two dimensional electrophoresis or directly via matrix-assisted laser desorption ionization time of flight mass spectroscopy. Direct dilution, protein precipitation, ultrafiltration, and solid phase extraction in combination to the above profiling technologies serve the means for reliable proteomics analysis of one of the most significant yet very complex biological samples. PMID- 18287773 TI - Combining laser capture microdissection and proteomics techniques. AB - Laser microdissection is an effective technique to harvest pure cell populations from complex tissue sections. In addition to using the microdissected cells in several DNA and RNA studies, it has been shown that the small number of cells obtained by this technique can also be used for proteomics analysis. Combining laser capture microdissection and different types of mass spectrometers opened ways to find and identify proteins that are specific for various cell types, tissues, and their morbid alterations. Although the combination of microdissection followed by the currently available techniques of proteomics has not yet reached the stage of genome wide representation of all proteins present in a tissue, it is a feasible way to find significant differentially expressed proteins in target tissues. Recent developments in mass spectrometric detection followed by proper statistics and bioinformatics enable to analyze the proteome of not more than 100-200 cells. Obviously, validation of result is essential. The present review describes and discusses the various methods developed to target cell populations of interest by laser microdissection, followed by analysis of their proteome. PMID- 18287774 TI - Comparison of protein expression by isotope-coded affinity tag labeling. AB - Isotope-coded affinity tag (ICAT) labeling, in combination with mass spectrometry (MS), has been widely adopted as an effective method for comparing protein abundance levels. This chapter describes the ICAT labeling procedure in search for the celecoxib-regulated proteins in a colon cancer cell line. Celecoxib, a cyclooxygenase-2 (COX-2) specific inhibitor, is used as a colorectal cancer preventative drug in clinical trials. Here, celecoxib is used to inhibit the expression of COX-2 in a colon cancer cell line HT-29. To elucidate the proteomic changes induced by celecoxib, the protein lysates from the treated and control cells are prepared. The cysteine-containing proteins are labeled with the heavy and light ICAT reagents, respectively. The labeled proteins are then combined and digested with trypsin. The ICAT-labeled peptides are subject to the purification through an avidin column and eventually the cleavage of the biotin tags. This chapter focuses on the ICAT labeling procedure itself, because sample preparation is the most critical step of an ICAT-based protein expression comparison experiment. Other related procedures such as the cation exchange high performance liquid chromatography separation of peptides and MS analysis are detailed elsewhere in this book. PMID- 18287775 TI - Analysis of microdissected cells by two-dimensional LC-MS approaches. AB - Laser capture microdissection (LCM) is a powerful tool that enables the isolation of specific cell types from tissue sections, overcoming the problem of tissue heterogeneity and contamination. We combined the LCM with isotope-coded affinity tag (ICAT) technology and two-dimensional liquid chromatography to investigate the qualitative and quantitative proteomes of hepatocellular carcinoma (HCC). The effects of three different histochemical stains on tissue sections have been compared, and toluidine blue stain was proved as the most suitable stain for LCM followed by proteomic analysis. The solubilized proteins from microdissected HCC and non-HCC hepatocytes were qualitatively and quantitatively analyzed with two dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) alone or coupled with cleavable isotope-coded affinity tag (cICAT) labeling technology. A total of 644 proteins were qualitatively identified and 261 proteins were unambiguously quantified. These results showed that the clinical proteomic method using LCM coupled with ICAT and 2D-LC-MS/MS can carry out not only large-scale but also accurate qualitative and quantitative analysis. PMID- 18287776 TI - Label-free LC-MS method for the identification of biomarkers. AB - Pharmaceutical companies and regulatory agencies are pursuing biomarkers as a means to increase the productivity of drug development. Quantifying differential levels of proteins from complex biological samples like plasma or cerebrospinal fluid is one specific approach being used to identify markers of drug action, efficacy, toxicity, etc. Academic investigators are also interested in markers that are diagnostic or prognostic of disease states. We report a comprehensive, fully automated, and label-free approach to relative protein quantification including: sample preparation, proteolytic protein digestion, LCMS/MS data acquisition, de-noising, mass and charge state estimation, chromatographic alignment, and peptide quantification via integration of extracted ion chromatograms. Additionally, we describe methods for transformation and normalization of the quantitative peptide levels in multiplexed measurements to improve precision for statistical analysis. Lastly, we outline how the described methods can be used to design and power biomarker discovery studies. PMID- 18287777 TI - Analysis of the extracellular matrix and secreted vesicle proteomes by mass spectrometry. AB - The extracellular matrix (ECM) and secreted vesicles are unique structures outside of cells that carry out dynamic biological functions. ECM is created by most cell types and is responsible for the three-dimensional structure of the tissue or organ in which they are originated. Many cells also produce or secrete specialized vesicles into the ECM, which are thought to influence the extracellular environment. ECM is not s a physical structure to connect cells in a tissue or organ. The proteins in ECM and secreted vesicles are critical to cell function, differentiation, motility, and cell-to-cell interaction. Although a number of major structural proteins of ECM and secreted vesicles have long been known, an appreciation of the role of less-abundant non-collagenous proteins has just begun to emerge. This chapter outlines a series of methods used to isolate and enrich ECM constituents and secreted vesicles from bone-forming osteoblast cells, enabling comprehensive profiles of their proteomes to be obtained by mass spectrometry. These methods can be easily adapted to study ECM and secreted vesicles in other cell types, primary cell cultures derived from animal models, or tissue specimens. PMID- 18287778 TI - Miniaturized parallelized sandwich immunoassays. AB - This chapter describes the development and use of bead-based miniaturized multiplexed sandwich immunoassays for focused protein profiling. Bead-based protein arrays or suspension microarrays allow simultaneous analysis of a variety of parameters within a single experiment. In suspension microarrays capture antibodies are coupled onto color-coded microspheres. The applications of suspension microarrays are described, which allow to analyze proteins present in different types of body fluids, such as serum or plasma, cerebrospinal, pleural and synovial fluids, as well as cell culture supernatants. The chapter is divided into the generation of suspension microarrays, sample preparation, processing of suspension microarrays, validation of analytical performance, and finally pattern generation using bioinformatics tools. PMID- 18287779 TI - Dissecting cancer serum protein profiles using antibody arrays. AB - Antibody arrays represent one of the high-throughput techniques enabling detection of multiple proteins simultaneously. One of the main advantages of the technology over other proteomic approaches resides on that the identities of the measured proteins are known at front of the experimental design or can be readily characterized, facilitating a biological interpretation of the obtained results. This chapter overviews the technical issues of the main antibody array formats as well as various applications using serum specimens in the context of neoplastic diseases. Clinical applications of antibody arrays vary from biomarker discovery for diagnosis, prognosis, and drug response to characterization of s protein pathways and modification changes associated with disease development and progression. As a high-throughput tool addressing protein levels and post translational modifications, it improves the functional characterization of molecular bases for cancer. Furthermore, the identification and validation of protein expression patterns characteristic of cancer progression and tumor subtypes may enable tailored therapeutic intervention and improvement in the clinical management of cancer patients. Technical requirements such as lower sample volume, antibody concentration, format versatility, and high reproducibility support their increasing impact in cancer research. PMID- 18287780 TI - 2D-PAGE maps analysis. AB - Due to the low reproducibility affecting 2D gel-electrophoresis and the complex maps provided by this technique, the use of effective and robust methods for the comparison and classification of 2D maps is a fundamental tool for the development of automated diagnostic methods. A review of classical and recently developed methods for the comparison of 2D maps is presented here. The methods proposed regard both the analysis of spot volume datasets through multivariate statistical tools (pattern recognition methods, cluster analysis, and classification methods) and the analysis of 2D map images through fuzzy logic, three-way PCA, and the use of moment functions. The theoretical basis of each procedure is briefly introduced, together with a review of the most interesting applications present in recent literature. PMID- 18287781 TI - Finding the significant markers: statistical analysis of proteomic data. AB - After separation through two-dimensional gel electrophoresis (2DE), several hundreds of individual protein abundances can be quantified in a cell population or sample tissue. Both a good experimental setup and a valid statistical approach are essential to get insight into the data and to draw correct conclusions. High throughput 2DE proteomics yield complex and large datasets with a huge disproportion between the hundreds of variables and the restricted number of replicates. However, the most commonly used statistical tests have been designed to cope with a high number of replicates and a restricted number of variables. There is some inconsistency in the proteomics community related to the use of statistics. Two approaches of data analysis can be distinguished: exploratory data analysis and confirmatory data analysis. Currently, most proteomic data are analyzed with the emphasis on confirmatory analysis and do not take into account the exploratory data analysis. This chapter gives an overview of the typical statistical exploratory and confirmatory tools available and suggests case specific guidelines for a reliable statistical approach that can be used for 2DE analysis. Examples are given for an experimental setup based on classical staining methods as well as for the more advanced difference gel electrophoresis. PMID- 18287782 TI - Web-based tools for protein classification. AB - Current proteomics technologies generate large number of data among which the investigator has to identify the promising diagnostic/prognostic biomarkers as well as potential therapeutic targets. For the latter, classification of proteins into meaningful families is needed. Current databases, featuring a high level of interconnectivity (cross referencing), provide the tools necessary to bring various data together, facilitating protein classification and elucidation of protein function and interoperativity. This chapter provides guidelines to explore the informationally rich peptide sequences generated by the application of the proteomics methodologies by the use of web-based tools, with the objective to predict potential protein function. After proper preprocessing (e.g., for internal repeats) of a query protein sequence, known domains can be identified, which aid in dividing the query into smaller meaningful parts. Any unclassified remainder of the protein provides the material for low-level comparative analysis for the discovery of distant homologues or candidate novel domain types to be verified experimentally. PMID- 18287783 TI - Open-source platform for the analysis of liquid chromatography-mass spectrometry (LC-MS) data. AB - The analysis of protein mixtures by liquid chromatography-mass spectrometry (LCMS) requires tools for viewing and navigating LC-MS data, locating peptides in LC-MS data, and eliminating low-quality peptides. msInspect, an open source platform, can carry out these steps for single experiments and can align and normalize peptide features in comparative studies with multiple LC-MS runs. In addition, msInspect can analyze quantitative studies with and without isotopic labels to generate peptide arrays. PMID- 18287784 TI - Pattern recognition approaches for classifying proteomic mass spectra of biofluids. AB - The statistical classification strategy we have developed for magnetic resonance, infrared, and Raman spectra for the analysis of biomedical data is discussed, particularly as it applies to proteomic mass spectra. A general discussion of the current use of pattern recognition methods is given, with caveats and suggestions relevant for clinical applicability. PMID- 18287787 TI - Mental ability in childhood and cognitive aging. AB - BACKGROUND: Identifying the determinants of cognitive aging is a research priority; however, few studies are able to examine the influence of pre-morbid cognitive ability on later changes in cognitive function. OBJECTIVE: To examine the association between childhood cognitive ability and cognitive change from age 79 to 83 in the presence of other demographic and lifestyle indicators. METHODS: The participants took a test of mental ability when aged 11 as part of the Scottish Mental Survey 1932. Cognitive ability based on Raven's Matrices, Verbal Fluency, and Logical Memory was assessed at ages 79 and 83. We used both linear regression and latent variable growth curve modeling to compare methods and results. RESULTS: Using linear regression, childhood mental ability was a significant predictor of cognitive change from 79 to 83, accounting for about 1.4% of the variance. Sex, education, social class, smoking status and alcohol intake were non-significant. In contrast, using latent variable growth curve modeling, there was no association between childhood mental ability and cognitive change. Sex (male), years of education, drinking status (positive), and childhood IQ were associated with better cognitive ability at age 79. The difference in results was due to the inability of linear regression to account completely for test-specific variance. CONCLUSION: Within a group of non-demented older people, greater childhood mental ability was associated with level of cognitive ability at age 79, but not with change in cognitive ability to age 83. To obtain accurate results regarding covariates of change, it is important to use methodology that can appropriately allocate all measured sources of variance. PMID- 18287786 TI - Inhibitory effect of a potent and selective cytosolic phospholipase A2alpha inhibitor RSC-3388 on skin inflammation in mice. AB - Cytosolic phospholipase A2alpha (cPLA2alpha) preferentially hydrolyzes membrane phospholipids containing arachidonic acid, resulting in the biosynthesis of eicosanoids such as prostaglandins and leukotrienes. To examine the contribution of cPLA2alpha to skin inflammation, we evaluated the effect of (E)-N-[(2S,4R)-4 [N-(biphenyl-2-ylmethyl)-N-2-methylpropylamino]-1-[2-(2,4 difluorobenzoyl)benzoyl]pyrrolidin- 2-yl]methyl-3-[4-(2,4-dioxothiazolidin-5 ylidenemethyl) phenyl]acrylamide (RSC-3388), a potent and selective cPLA2alpha inhibitor, on 2,4,6-trinitro-1-chlorobenzene (TNCB)-induced ear inflammation and mite antigen-induced dermatitis in mice. Topical application of RSC-3388 showed a significant inhibitory activity against TNCB-induced ear swelling and eicosanoid production in mice. Comprehensive expression analysis using Gene-Chip technology and subsequent experiments concerning mRNA and protein expression demonstrated that RSC-3388 clearly reduced the levels of interleukin-1beta, macrophage inflammatory protein-1alpha (MIP-1alpha) and MIP-1beta in a TNCB-induced mouse model. In addition, RSC-3388 ointment significantly alleviated atopic dermatitis like skin lesions induced by repeated application of mite antigen. Furthermore, increased expression of cPLA(2)alpha, assessed by anti-phospho-cPLA2alpha antibody, was observed in the skin lesions of mite-antigen-induced dermatitis. These results indicate that cPLA2alpha is involved in the development of skin inflammation in mice, and RSC-3388 is expected to be useful for the treatment of inflammatory skin disorders such as atopic dermatitis. PMID- 18287788 TI - Electromyographic activity of the mandible muscles at the beginning and end of masticatory cycles in patients with complete dentures. AB - The aim of this study was to assess the electrical activity of the masseter and anterior temporal muscles in subjects with severe bone resorption at two different times: (a) the initial period, with the complete dentures they had worn for over 10 years, and (b) the final period, 5 months after having new dentures put into place. Twelve asymptomatic subjects were asked to respond to the questionnaire, according to the research diagnostic criteria for temporomandibular disorders, before denture rehabilitation and 5 months after the new dentures were put in place. The electrical activity of the muscles was recorded during mastication in the initial and final period, using artificial food (Optocal). The operator monitored the 35 chewing cycles that were repeated to grind the artificial food. After wearing the new dentures for 5 months, the right anterior temporal muscle showed a statistical difference before and after denture rehabilitation at the beginning and end of mastication. Muscular capacity and ability reduced the electrical activity in the masseter muscles after rehabilitation. PMID- 18287790 TI - Effect of different concentrations of fluoride in dentifrices on dentin erosion subjected or not to abrasion in situ/ex vivo. AB - This in situ/ex vivo study assessed the effect of different concentrations of fluoride in dentifrices on dentin subjected to erosion or to erosion plus abrasion. Ten volunteers took part in this crossover and double-blind study performed in 3 phases (7 days). They wore acrylic palatal appliances containing 4 bovine dentin blocks divided in two rows: erosion and erosion plus abrasion. The blocks were subjected to erosion by immersion ex vivo in a cola drink (60 s, pH 2.6) 4 times daily. During this step, the volunteers brushed their teeth with one of three dentifrices D (5,000 ppm F, NaF, silica); C (1,100 ppm F, NaF, silica) and placebo (22 ppm F, silica). Then, the respective dentifrice slurry (1:3) was dripped on dentin surfaces. While no further treatment was performed in one row, the other row was brushed using an electric toothbrush for 30 s ex vivo. The appliances were replaced in the mouth and the volunteers rinsed with water. Dentin loss was determined by profilometry and analyzed by 2-way ANOVA/Bonferroni test (a = 0.05). Dentin loss after erosive-abrasive wear was significantly greater than after erosion alone. Wear was significantly higher for the placebo than for the D and C dentifrices, which were not significantly different from each other. It can be concluded that the presence of fluoride concentrations around 1,100 ppm in dentifrices is important to reduce dentin wear by erosion and erosion + abrasion, but the protective effect does not increase with fluoride concentration. PMID- 18287789 TI - Pulse wave velocity is associated with metabolic syndrome components in CAPD patients. AB - BACKGROUND: Overall, the proportion of deaths due to cardiovascular disease (CVD) reached 40-50% in dialysis patients. Pulse wave velocity (PWV) reflects arterial stiffness and may provide an integrated index of vascular status and CVD risk. Individual components of the metabolic syndrome (MetS) are well-established cardiovascular risk factors. Thus we conducted a cross-sectional study in continuous ambulatory peritoneal dialysis (CAPD) patients to explore the association of MetS components with PWV. METHODS: Prevalent CAPD patients (n = 148, 63 M/85 F) were categorized according to the number of traits of the MetS into one of three groups (No MetS, Risk of MetS, MetS). Due to the effect of peritoneal dialysis (PD), waist circumference was not assessed. Aortic stiffness was assessed by carotid-femoral PWV (C-F PWV). RESULTS: Patients' MetS trait numbers were positively associated with C-F PWV (r = 0.301, p < 0.01), pulse pressure (r = 0.256, p < 0.01), systolic blood pressure (r = 0.233, p < 0.01), and serum albumin (r = 0.205, p < 0.05). In a multivariate regression analysis, PWV was independently determined by age (p < 0.01) and MetS score (p = 0.01). Adjusted R(2) of the model was 0.24. CONCLUSION: MetS traits were closely associated with an increased C-F PWV, even after adjustment for confounders. This suggests that commonly recognized MetS criteria are useful also when predicting CVD in CAPD patients. PMID- 18287791 TI - Nanomedicine: application of nanobiotechnology in medical practice. AB - Nanomedicine is the application of nanobiotechnologies to medicine. This article starts with the basics of nanobiotechnology, followed by its applications in molecular diagnostics, nanodiagnostics, and improvements in the discovery, design and delivery of drugs, including nanopharmaceuticals. It will improve biological therapies such as vaccination, cell therapy and gene therapy. Nanobiotechnology forms the basis of many new devices being developed for medicine and surgery such as nanorobots. It has applications in practically every branch of medicine and examples are presented of those concerning cancer (nanooncology), neurological disorders (nanoneurology), cardiovascular disorders (nanocardiology), diseases of bones and joints (nanoorthopedics), diseases of the eye (nanoophthalmology), and infectious diseases. Safety issues of in vivo use of nanomaterials are also discussed. Nanobiotechnology will facilitate the integration of diagnostics with therapeutics and facilitate the development of personalized medicine, i.e. prescription of specific therapeutics best suited for an individual. Many of the developments have already started and within a decade a definite impact will be felt in the practice of medicine. PMID- 18287792 TI - Gender differences in pharmacological and clinical associates of kidney disease. A hospital-based study. AB - OBJECTIVE: The objective of this study was to identify the pharmacological and clinical associates of kidney disease among males and females. SUBJECTS AND METHODS: This is a cross-sectional retrospective study. All patients admitted to the Department of Internal Medicine at Al-Watani Hospital were included in the study. Medical data were obtained from patients' medical records. Kidney disease was defined as a glomerular filtration rate (GFR) less than the published cutoff points. Males with GFR <64.25 ml/min and females with GFR <59.25 ml/min were designated as kidney group and those above were considered the reference group. GFR was estimated using Cockroft-Gault equation; chi(2) and multiple logistic regressions were used to test for significance using statistical package for social sciences program. RESULTS: Of the 340 patients, 185 (54.5%) were males and 155 (45.6%) females; mean age was 60.4 +/- 17 years. Of the 185 males, 58 (31.4%) and of the 155 females, 41 (26.6%) had low GFR kidney disease. Stepwise forward multiple logistic regression in males identified age, hypertension, and diabetes mellitus (DM) as associates of low GFR. In females, DM and angiotensin-converting enzyme inhibitors (ACE-I)/diuretic utilization were identified as significant associates of having low GFR. As the number of clinical conditions present in the patient increased, the odds ratio of having low GFR increased, in a similar and parallel way in both genders. CONCLUSION: Diabetes mellitus, but not hypertension or advancing age, was significantly associated with low GFR in both males and females. Utilization of the ACE-I/diuretic combination was significantly associated with low GFR among females but not males. Patients, regardless of gender, who had multiple clinical conditions, had higher risk of having low GFR. PMID- 18287793 TI - Indications of the mechanisms involved in improved sperm parameters by zinc therapy. AB - OBJECTIVE: To determine possible indications of the mechanisms involved in improved sperm parameters by zinc therapy in asthenozoospermic men. SUBJECTS AND METHODS: Forty-five men with asthenozoospermia (>or=40% immotile sperm) were randomized into four therapy groups: zinc only: n = 11; zinc + vitamin E: n = 12 and zinc + vitamins E + C: n = 14 for 3 months, and non-therapy control group: n = 8. Semen analysis was done according to WHO guidelines. Malone dialdehyde, tumour necrosis factor-alpha (TNF-alpha), total antioxidant capacity, superoxide dismutase (SOD) and glutathione peroxidase were determined in the semen and serum. Antisperm antibodies IgG, IgM and IgA were evaluated by immunobeads. Sperm chromatin integrity was determined by acid denaturation by acridine orange and sperm apoptosis by light and electron microscopy. The effect of zinc on in vitro induced sperm oxidative stress by NADH was evaluated. RESULTS: Asthenozoospermia was significantly associated with oxidative stress with higher seminal malone dialdehyde (8.8 vs. 1.8 mmol/l, p < 0.001) and TNF-alpha (60 vs. 12 pg/l, p < 0.001), and low total antioxidant capacity (1.8 vs. 8.4, p < 0.01), SOD (0.8 vs. 3.1, p < 0.01) and glutathione peroxidase (1.6 vs. 4.2, p < 0.05), compared to normozoospermia. Zinc therapy alone, in combination with vitamin E or with vitamin E + C were associated with comparably improved sperm parameters with less oxidative stress, sperm apoptosis and sperm DNA fragmentation index (DFI). On the whole, there was no difference in the outcome measures between zinc only and zinc with vitamin E and combination of vitamins E + C. In the in vitro experiment zinc supplementation resulted in significantly lower DFI (14-29%, p < 0.05) compared to zinc deficiency. CONCLUSION: Zinc therapy reduces asthenozoospermia through several mechanisms such as prevention of oxidative stress, apoptosis and sperm DNA fragmentation. PMID- 18287794 TI - Effect of chemosensitizers on minimum inhibitory concentrations of fluconazole in Candida albicans. AB - OBJECTIVE: To evaluate the effect of chemosensitizers on the in vitro activity of fluconazole against Candida albicans strains. MATERIALS AND METHODS: Using Clinical Laboratory Standard Institute method, antifungal activity of fluconazole was determined alone and in combination with 16 chemosensitizers that included verapamil, reserpine, quinine, quinidine, gemfibrozil, lansoprazole, tamoxifen, diltiazem, desipramine, nicardipine, cyclosporine, chlorpromazine, prochlorperazine, promethazine, thioridazine, and trifluoperazine. Further studies were done using double combinations of selected chemosensitizers with fluconazole (28 combinations). For testing combinations, half of the minimum inhibitory concentration (MIC) of each agent was selected in order to avoid the effect of the drug alone. One reference strain (ATCC90028) and one clinical isolate of C. albicans were used for testing the in vitro activity. Broth dilution method was used to determine the MICs of fluconazole and chemosensitizers. RESULTS: Of the 16 chemosensitizers tested, 3 exhibited in vitro activity by increasing fluconazole susceptibility to 7-fold. The MICs of the reference strain and clinical isolate for fluconazole were 5.5 and 0.55 microg/ml, respectively, and these were reduced to 0.76 microg/ml by gemfibrozil, 0.83 microg/ml by quinine, and 0.76 microg/ml by chlorpromazine in the reference strain, with MIC reduction to 0.08 microg/ml by all three chemosensitizers in the clinical isolate. Some double combinations reduced the MIC of fluconazole to 10- to 100-fold, even when the chemosensitizers were not effective alone. CONCLUSION: The most effective double combinations were those of chlorpromazine with either reserpine or nicardipine. PMID- 18287795 TI - Aseptic meningitis among children in Kuwait. AB - OBJECTIVE: To describe the epidemiology, clinical features, and the morbidity caused by aseptic meningitis in children in Kuwait. SUBJECTS AND METHOD: A multicenter retrospective study of previously healthy children hospitalized with a diagnosis of aseptic meningitis in the period 2001-2003 was carried out. RESULTS: There were 172 children with the diagnosis of meningitis based on changes in the cerebrospinal fluid. Aseptic meningitis was diagnosed in 86 (50%) of the cases; their mean age was 3.2 +/- 3.8 years. There were two peaks of cases, one during the months of May and June and the other in November/December. Older children (>or=5 years) presented more frequently with signs and symptoms suggestive of meningeal irritation than younger children (<5 years) (p 0.05). CONCLUSION: Our results showed that smoking affected white blood cell count and von Willebrand factor levels in males and not in females, and as such could be potential markers for smoking-induced endothelial damage in asymptomatic Arab male smokers. PMID- 18287801 TI - Primary mediastinal pleomorphic rhabdomyosarcoma: a case report. AB - OBJECTIVE: To report a rare case of pleomorphic rhabdomyosarcoma which occurred in the mediastinum of a 34-year-old man. CLINICAL PRESENTATION AND INTERVENTION: A young male labourer presented with dyspnoea on exertion. A large mediastinal mass was detected on chest CT scan. The chest surgeons advised against open biopsy. His alpha-fetoprotein was 22,000 IU/l; based on this the diagnosis of a germ cell tumour was made and the patient was treated with a bleomycin/etoposide/cisplatin regimen. He left for his native country where an open biopsy from the mediastinum was taken and reported as pleomorphic rhabdomyosarcoma. He was given five courses of chemotherapy with doxorubicin, etoposide, and ifosfamide with mesna protection without much relief. The inoperable disease occupied the whole of the right chest and mediastinum. The enormous size of the radiation field made radiotherapy prohibitive. Finally, the patient opted for symptomatic treatment and left for his native place. CONCLUSION: This case is presented because of its difficulty in management and rarity. PMID- 18287802 TI - Acute coronary stenosis in a young man with Behcet's syndrome. AB - OBJECTIVE: To present a case of myocardial infarction due to Behcet's syndrome. CLINICAL PRESENTATION AND INTERVENTION: A 27-year-old man who was known to have Behcet's syndrome for 1 year presented with retrosternal fluctuating chest pain, which radiated to the epigastrium 5 h prior to admission. Coronary angiography showed total occlusion of the left anterior descending coronary artery, which was successfully treated with coronary stent implantation. CONCLUSION: This case shows that patients with Behcet's syndrome who had acute chest pain should be thoroughly examined for any signs of acute myocardial infarction. PMID- 18287803 TI - Ruptured Meckel's mesodiverticulum and Meckel's diverticulum following blunt abdominal trauma. AB - OBJECTIVES: To present a case of simultaneous rupture of Meckel's diverticulum and mesodiverticulum with abdominal pain following a blunt trauma to the abdomen, sustained during an automobile accident. CLINICAL PRESENTATION: Following a head on automobile collision a 36-year-old man was referred to the emergency room with abdominal pain, guarding and rigidity and was taken to the operating theater with a preoperative impression of peritonitis due to rupture of a hollow viscus. Upon abdominal exploration, ruptured Meckel's diverticulum (from the base) and ileum with active bleeding from mesodiverticulum was found. INTERVENTION: The ileum was repaired in two layers: a segmental bowel resection including Meckel's diverticulum and the gastrointestinal tube anastomosed without any tension. The patient has a normal postoperative course. CONCLUSION: This case shows that a blunt abdominal trauma can tear the mesodiverticulum and rupture the Meckel's diverticular base simultaneously, resulting in hemoperitoneum and chemical peritonitis. Diagnosis and management of this type of injury are basically the same as those of acute surgical abdomen following trauma. PMID- 18287804 TI - Diffuse cystic bronchiectasis associated with left microtia and external auditory canal atresia: a very rare coincidence. AB - OBJECTIVE: To present a case in which diffuse cystic bronchiectasis was associated with left microtia/external auditory canal atresia. CLINICAL PRESENTATION: A 10-year-old girl suffering from cough, fever, dyspnea and sputum for 6 months was transferred to our clinic due to the diagnosis of bronchopneumonia. She had recurrent episodes of bronchopneumonia. On examination, left microtia and left external auditory canal atresia were detected. Thorax CT revealed diffuse cystic bronchiectasis on the left lung. Radiological examination showed atresia of the left external auditory canal. We could not find any etiopathological reason causing bronchiectasis. INTERVENTION: The patient underwent left pneumonectomy and postoperative histopathology was reported as bronchiectasis. CONCLUSION: This report shows a unique case in which an association of diffuse cystic bronchiectasis and left microtia/external auditory canal atresia was observed. Hence in newborns with microtia and/or external auditory canal atresia, the probability of development of bronchiectasis should be borne in mind and such patients should be followed up more carefully regarding this rare association. PMID- 18287805 TI - The use of lithium carbonate in the preparation for definitive therapy in hyperthyroid patients. AB - OBJECTIVE: The aim of this study was to elucidate the effectiveness of lithium carbonate prior to thyroidectomy or radioiodide therapy in patients with thyrotoxicosis. SUBJECTS AND METHODS: Lithium carbonate was used as preoperative preparation or radioiodide therapy in 5 patients with Graves' disease and in 1 patient with toxic multinodular goiter because of side effects of thionamide in 5 patients and ineffectiveness of antithyroid medication in the remaining patient. RESULTS: All 6 patients had a benign course following treatment without thyroid storm. No adverse effects or complications of lithium carbonate were observed. CONCLUSION: This report shows that lithium carbonate can be safely used preoperatively or prior to radioiodide therapy in circumstances where antithyroid medications are contraindicated and are ineffective in obtaining an euthyroid status. PMID- 18287806 TI - Problem-based learning: the right direction for medical teaching? PMID- 18287807 TI - Urinary IL-18 and NGAL as early predictive biomarkers in contrast-induced nephropathy after coronary angiography. AB - BACKGROUND/AIMS: Contrast-induced nephropathy (CIN) is at present the third leading cause of hospital-acquired acute kidney injury (AKI). Traditionally, it is diagnosed by measuring the increase of the serum creatinine concentration. However, in patients with acute changes in their glomerular filtration rate, serum creatinine is an insensitive marker. This clinical study was designed to investigate whether human urinary interleukin-18 (IL-18) and neutrophil gelatinase-associated lipocalin (NGAL) are early predictive markers for AKI after coronary angiography and their correlation with later cardiac events. METHODS: Patients undergoing coronary angiography using low-osmolar contrast medium were enrolled and then followed up for at least 17 months. Urine samples were collected before and 24 h after coronary angiography and IL-18 and NGAL levels measured by using an ELISA kit. RESULTS: CIN was diagnosed in 13 of 150 (8.7%) patients (CIN group); 27 patients without CIN served as control group. At 24 h after the procedure, the urinary IL-18 and NGAL levels were significantly increased in the CIN group, but not in the control group (p < 0.05). The predictable time of AKI onset determined by IL-18 was 24 h earlier than determined by serum creatinine (p < 0.01). Receiver operating characteristic curve analysis showed that both IL-18 and NGAL showed a good performance in early diagnosis of CIN as compared with serum creatinine (p < 0.05). We also found that IL-18 is an independent predictive marker for later major cardiac events: relative risk = 2.09 (p < 0.01). CONCLUSIONS: We conclude that urinary IL-18 or NGAL could be early biomarkers of CIN and that urinary IL-18 is well associated with the later cardiac outcomes in patients after coronary angiography. PMID- 18287808 TI - Novel SLC12A3 mutations in Chinese patients with Gitelman's syndrome. AB - BACKGROUND: Inactivating mutations of the SLC12A3 gene are the most common cause of Gitelman's syndrome (GS), a disorder inherited as an autosomal recessive trait. In a minority of cases, GS-like phenotypes are caused by mutations in the CLCNKB gene. METHODS: We searched for SLC12A3 and CLCNKB gene mutations in 13 Chinese patients (9 males and 4 females, age 35 +/- 14 years) from 8 unrelated families with the clinical and biochemical features of GS. All coding regions, including intron-exon boundaries, were analyzed using PCR followed by direct sequence analysis. RESULTS: We identified 10 mutations distributed throughout the SLC12A3 gene. Seven are novel variants, including 4 missense mutations (Gly196Val, Cys430Gly, Gly439Val and Leu571Pro), 2 deletions (1384delG and 346 353delACTGATGG) and 1 in-frame insertion (997insCys). Three mutations were recurrent, including 2 missense mutations (Thr60Met and Asp486Asn) and 1 deletion (2883-2884delAG). The homozygous or heterozygous mutation Thr60Met was found in 8 of 13 patients. There were no mutations detected in the CLCNKB gene. CONCLUSIONS: Thr60Met may be the most common mutation in Chinese patients with GS. Possible specific genotype-phenotype correlations were difficult to identify. PMID- 18287809 TI - Activation of human T cells induces upregulation of cannabinoid receptor type 1 transcription. AB - OBJECTIVE: Effects of cannabinoids are mediated by CB1 and CB2 receptors. In addition to neuronal effects, cannabinoids are potent modulators of immune functions. In this report, we investigated whether the transcription of these receptors is regulated after activation of T lymphocytes. METHODS: CB1- and CB2 specific mRNA of primary human peripheral blood T cells and cells of the human T cell line Jurkat was measured by quantitative real-time RT-PCR in response to CD3/28. Using the decoy oligonucleotide approach, transcription factors involved in the regulation were determined. A promoter analysis was performed using transient transfection of chloramphenicol acetyl transferase reporter gene constructs in Jurkat cells. RESULTS: Activation of human T cells caused an induction of CB1 mRNA expression in primary human T cells (8-fold) and Jurkat cells (29-fold). In contrast, CB2 transcription was not regulated. The CD3/28 mediated upregulation of CB1 involves the transcription factors AP-1, NF kappaB and NFAT. Furthermore, 2,490 bp of the CB1 promoter mediated inducibility in response to CD3/28. CONCLUSIONS: The upregulation of CB1 in activated T cells, together with the constitutive expression of CB2, enables cellular responses to cannabinoids mediated by both receptor subtypes. It may thus contribute to the understanding of the various modulatory effects of cannabinoids on activated T cells. PMID- 18287810 TI - Stress and prolactin effects on bone marrow myeloid cells, serum chemokine and serum glucocorticoid levels in mice. AB - OBJECTIVE: Current evidence supports the conclusion that prolactin (PRL) is not an obligate immunoregulatory hormone and influences the immune system predominantly during stress conditions. In this study, we examined the impact of PRL on the psychogenic stress-induced responses of myeloid cells. METHODS: Seven week-old PRL+/- (normal) and PRL-/- (deficient) mice were exposed to a predator for 1 h/day on 3 consecutive days. Another group of PRL-deficient mice received either 1 pituitary graft (hyperprolactinemic) or sham surgery at 5 weeks of age, while PRL-normal mice only received sham surgery. Two weeks later, these mice were also subjected to predator exposure. One day after the last predator exposure session, all mice were killed and the bone marrow and blood harvested. RESULTS: Significant differences in the myeloid cells between PRL-normal and PRL deficient mice only occurred in stressed conditions. The median serum corticosterone levels were consistently higher in PRL-deficient mice. The implantation of a pituitary graft lowered the corticosterone levels to those observed in PRL-normal mice. The absolute number of immature neutrophils as well as the numbers of granulocyte macrophage, monocyte/macrophage and granulocyte colonies were significantly higher in the stressed PRL-deficient mice; however, only the increased number of immature neutrophils was reversed by pituitary grafting. CONCLUSIONS: Our findings support previous observations that PRL influences myeloid cells of the bone marrow most profoundly in stressed conditions. However, the mechanism by which PRL influences bone marrow myeloid cells during stress cannot be explained solely by its effect on serum corticosterone. PMID- 18287812 TI - Characterization of the activation domain of the Rad53 checkpoint kinase. AB - Rad53 protein, the yeast orthologue of the human checkpoint kinase Chk2, presents two highly conserved phosphorylatable threonine residues (T354 and T358) in the activation domain, whose phosphorylation is critical to allow the activation of the kinase. In this study we found that Rad53 protein variants in which alanine and/or aspartate replace the threonine residues 354 and/or 358 do not retain kinase activity and do not undergo auto-phosphorylation, leading to defect in the checkpoint response and iper-sensitivity to DNA damage and DNA replication stress agents. Interestingly, we found that the rad53-T358D mutation severely affects the kinase activity and causes accumulation of the S129-phosphorylated isoform of histone H2A, even during an unperturbed cell cycle, thus indicating the accumulation of spontaneous DNA breaks. We further found that the protein level of Sml1, which is the physiological inhibitor of ribonucleotide reductase, remains high during DNA replication in rad53-T358D cells, suggesting that an inadequate pool of dNTPs in checkpoint defective cells causes the accumulation of spontaneous DNA breaks. In conclusion, our results indicate that phosphorylation of both T354 and T358 residues strongly influences the catalytic activity of Rad53 also in unperturbed cell cycles, and support the notion that Rad53 is essential to preserve genome integrity, by controlling the level of Sml1 and the functionality of ribonucleotide reductase. PMID- 18287811 TI - Integrin affinity modulation in angiogenesis. AB - Integrins, transmembrane glycoprotein receptors, play vital roles in pathological angiogenesis, but their precise regulatory functions are not completely understood and remain controversial. This study aims to assess the regulatory functions of individual beta subunits of endothelial integrins in angiogenic responses induced by vascular endothelial growth factor (VEGF). Inhibition of expression of beta(1), beta(3), or beta(5) integrins in endothelial cells resulted in down regulation of EC adhesion and migration on the primary ligand for the corresponding integrin receptor, while no effects on the recognition of other ligands were detected. Although inhibition of expression of each subunit substantially affected capillary growth stimulated by VEGF, the loss of beta(3) integrin was the most inhibitory. PMID- 18287813 TI - Genetic association of LOXL1 gene variants and exfoliation glaucoma in a Utah cohort. AB - Exfoliation glaucoma (XFG) is the commonest identifiable cause of secondary open angle glaucoma worldwide, characterized by the deposition of fibrillar proteins in the anterior segment of the eye. We investigated LOXL1 gene variants previously identified to confer susceptibility to XFG in a Utah Caucasian cohort. After a standard eye examination protocol we genotyped SNPs rs2165241and rs3825942 in 62 XFG or exfoliation syndrome (XFS) patients and 170 normal controls. Genotype frequency distribution, odds ratios (ORs) and population attributable risks were calculated for the risk alleles. The SNP rs2165241 was significantly associated with XFG and XFS (p = 4.13 x 10(-9)) for an additive model, OR(het) = 4.42 (2.30-8.50), OR(hom) = 34.19 (4.48-261.00); T allele: 83.1% in cases versus 52.4% in controls). Significant association was also found for rs3825942: (p = 1.89 x 10(-6)). Our findings confirm genetic association of LOXL1 with XFG and XFS and implicate a potential role of cross linking of elastin in the pathogenesis of XFG. This information will potentially guide glaucoma monitoring efforts by targeting individuals whose genetic profiles put them at higher risk for XFG. PMID- 18287814 TI - Effect of dietary tetradecylthioacetic acid on colon cancer growth studied by dynamic contrast enhanced MRI. AB - PURPOSE: To evaluate the effects of diets supplemented with the modified fatty acid tetradecylthioacetic acid (TTA) and fish protein hydrolysate (FPH) on tumor growth of the human colon cancer cell line SW620, and to investigate the properties of tumor vasculature by dynamic contrast-enhanced MRI in a human tumor xenograft. EXPERIMENTAL DESIGN: SW620 cells were grown in vitro in presence of TTA and palmitic acid and proliferation was measured by thymidine incorporation. The xenograft study in mice was performed with four distinct diets: (a) control diet; (b) diet with TTA; (c) diet with TTA and FPH; and (d) diet with FPH. SW620 cells were injected subcutaneously, and dynamic contrast enhanced (DCE) MRI was performed on a Bruker BioSpec 7T system. The data was analyzed by two-compartment modeling of the contrast enhancement, initial area under the curve (IAUC) and by use of relative signal intensity (RSI) distributions. RESULTS: The in vitro cell studies revealed that TTA reduced tumor cell proliferation as a function of both dose and time. The in vivo tumor growth was significantly reduced for the two groups fed TTA, as compared to the control group. The mean 10(th) percentile RSI, v(e) and IAUC for the TTA group were significant higher than for the control group. CONCLUSIONS: This study confirms the growth inhibitory effects of TTA, both in vitro and in vivo, in a colon cancer model. The analysis of DCE-MRI data showed that TTA influences the vascular properties of the tumor in addition to the growth. PMID- 18287815 TI - Center-of-pressure displacement during postural changes in relation to pressure ulcers in spinal cord-injured patients. AB - OBJECTIVE: To evaluate the center-of-pressure displacement in spinal cord-injured patients, to investigate dynamic sitting stability and its relationship with pressure ulcers. DESIGN: Sixteen spinal cord-injured patients and 18 healthy volunteers were included in the study. For the assessment of dynamic sitting stability, center-of-pressure displacement during maximum unsupported forward, backward, and right- and left-sided trunk leaning were measured with a seat sensor system, which was placed between the subject's buttocks and chair. RESULTS: Center-of-pressure displacements in all directions in spinal-injured patients were smaller than healthy volunteers (P < 0.05). Center-of-pressure displacements for high- and low-thoracic spinal cord-injured patients were not significantly different. History of previous pressure ulcer was not different between high- and low-thoracic spinal cord-injured patients (chi = 0.90, P = 0.62). Mean center-of-pressure displacement during forward and backward leaning were smaller in patients with pressure ulcer history (P = 0.04 and 0.03, respectively). CONCLUSIONS: The results of this study suggest that impaired dynamic sitting stability is associated with pressure ulcer development. PMID- 18287816 TI - Effect of interelectrode distance on sural nerve action potential parameters. AB - OBJECTIVE: To assess differences in amplitude, latency, and duration, using a 3 cm vs. 4-cm distance between the active and reference electrodes when performing sural nerve conduction studies (NCS). Current normative data in lower-limb studies are generally based on 3-cm interelectrode differences, although 4-cm differences have been reported to be optimal in the upper limb. DESIGN: Prospective study comparing the onset latency, peak latency, duration, and amplitude for the sural sensory nerve action potential (SNAP) recording at two interelectrode distances in adult volunteers. RESULTS: Forty-three sural nerves were studied in 22 normal subjects. Peak latencies recorded with a 4-cm interelectrode distance were significantly longer than those recorded with a 3-cm distance (mean difference = 0.06 msecs [SD = 0.09, P = 0.0073]). Duration was significantly longer (mean difference = 0.03 msecs [SD = 0.07, P = 0.0270]), conduction velocities were significantly slower (mean difference = -0.7 msecs [SD 1.0, P = 0.0012]), and onset latency and amplitude were not found to differ significantly. Average differences in peak latencies, duration, velocity, onset latency, and amplitude were not correlated with gender, age, or BMI. CONCLUSIONS: In contrast to studies of upper-limb sensory NCS, sural SNAP parameters obtained with 3- and 4-cm interelectrode distances did not differ for onset latencies and amplitude. Peak latencies, duration, and conduction velocity differences, though statistically significant, were of insufficient magnitude to be clinically meaningful. By using a 4-cm instead of a 3-cm interelectrode difference for sural nerve studies, the small prolongation of 0.06 msecs in peak latency and tiny increment of 0.27 muV observed in our investigation is unlikely to influence the electrodiagnostician's interpretation of the study. PMID- 18287817 TI - Effect of a high-rolling-resistance training method on the success rate and time required to learn the wheelchair wheelie skill: a randomized controlled trial. AB - OBJECTIVE: The primary objective of this study was to test the hypotheses that a wheelie training method that begins in a high-rolling-resistance (RR) setting (1) improves the success rate and (2) reduces the training time. Our secondary objectives were to assess the effects of other factors (e.g., age, gender) on training time and to acquire data on the perceptions of the participants that would assist us in refining our training methods. DESIGN: Randomized controlled study including 48 able-bodied participants with no significant wheelchair experience. Each participant was taught the wheelie skill, using up to five training sessions. Participants in the conventional group did all of their training on a smooth, level, tile surface. The RR group began training in a high RR setting (rear wheels initially prevented from moving at all, progressing to being on 12-cm-thick foam that permitted some movement) before moving to the tile surface. Outcome measures were success rate (%), training time (mins) to achieve wheelie competence (defined as the ability to perform two consecutive 30-sec wheelies within a 1.5-m-diameter circle, assessed at least 2 days after training), and a questionnaire. RESULTS: The success rates for those in the conventional and RR groups were 96% and 100%, respectively (P = 1.000). The mean (+/-SD) training times for the conventional and RR groups were 55.9 mins (+/ 27.1) and 51.8 mins (+/-18.7) (P = 0.549). Training time was not significantly affected by trainer or age, but it was affected by sex, with women requiring an average of 21.4 mins more than men (P = 0.002). Perceptions of participants in both groups about the training were positive. Of the participants in the RR group, 14 (74%) perceived the RR technique as "very effective," and five (26%) perceived it as "moderately effective." CONCLUSIONS: Neither success rate nor training time for wheelie skill acquisition by able-bodied learners are improved by a training method using high RR. Women require more time to learn than men. Learners using the RR technique perceive it to be effective. These results have implications for training practices. PMID- 18287818 TI - Publishing in physical and rehabilitation medicine. AB - This article presents the panel discussion from the "Meet the Editor" symposium held at the 4th World Congress of the International Society of Physical and Rehabilitation Medicine in Seoul in June 2006. It includes contributions by four Editors of international journals in rehabilitation. Some of the topics discussed are of a general nature but will provide useful guidance for the more junior scientific author. Some specific information about the four journals is also presented. Topics discussed include the reasons for publishing in peer-reviewed journals, important considerations in submitting a manuscript, the peer-review process, the effect of electronic publishing (which leads to shorter publication times and the opportunity to preview articles), and the trend toward more open access to journals. The discussion concludes that the field of physical and rehabilitation medicine will continue to expand, with an audience with a broader range of scientific and clinical interests. The International Classification of Functioning, Disability and Health (ICF) may be increasingly used as a framework in reporting. New journals may be started, particularly in regions of the world other than Europe and the United States, despite the fact that journals currently published in these regions are distributed worldwide. PMID- 18287819 TI - Association between the EMG self-assessment examination and later ABEM certification. AB - Our objective was to determine whether performance on the American Association of Neuromuscular and Electrodiagnostic Medicine Training Program Self-Assessment Examination (TPSAE) predicts subsequent performance on the American Board of Electrodiagnostic Medicine (ABEM) certification examination. The TPSAE results were closely associated (r = 0.57) with ranking on the ABEM examination. Those with scores <65% on the TPSAE had <50% chance of ABEM certification, whereas those with scores >84% correct had >90% chance of certification. The TPSAE is a useful predictor of later ABEM certification. PMID- 18287820 TI - Deep gray matter lesion caused by hypoglycemia. PMID- 18287822 TI - Overlap between orofacial pain and vulvar vestibulitis syndrome. AB - OBJECTIVES: To explore the prevalence of orofacial pain (OFP) among patients with vulvar vestibulitis syndrome (VVS) and to examine the relationship between signs and symptoms of OFP and clinical characteristics of women with VVS, we investigated differences in psychologic characteristics and severity of painful intercourse. METHODS: In this cross-sectional exploratory study, 137 women with VVS completed questionnaires that assessed levels of pain, anxiety, somatization, and presence of signs and symptoms suggestive of clinical and subclinical OFP. Demographic data were gathered from medical records. RESULTS: OFP was found to be a highly prevalent (78%) condition among women with VVS. Compared with women who had no OFP symptoms (n=30), those with symptoms (n=64) reported higher levels of anxiety (45.0 vs. 37.8, Bonferroni adjusted P=0.017), somatization (125.2 vs. 96.0, Bonferroni adjusted P<0.001), and psychologic distress (62.8 vs. 56.0, Bonferroni adjusted P=0.002). Although we observed a similar trend among women with subclinical OFP (n=43), this trend only reached statistical significance with respect to somatization. Differences were not detected for demographics, duration of pain, and severity of pain during intercourse across the 3 groups. DISCUSSION: OFP is a common condition among women with VVS. Because severity and duration of painful intercourse did not differ by OFP classification but psychologic characteristics did, we must begin to question a unidimensional focus on vestibular mucosa as a reason for pain and persistent distress. PMID- 18287823 TI - Response to morphine in male and female patients: analgesia and adverse events. AB - BACKGROUND: There is little agreement about a differential response of men and women to opioid analgesics. Some experimental and clinical studies have shown that women have a better response to opioids, others have found no difference, and still others have found opioids to be more effective analgesics for men than women. OBJECTIVES: To assess sex differences in analgesic response to morphine and incidence of adverse events in patients receiving a dose of 0.1 mg intravenous morphine/kg. METHODS: Secondary analysis of the control arms of 6 randomized clinical trials that compared 0.1 mg/kg intravenous morphine with other opioids or other doses of morphine in patients aged 21 to 65 with acute pain. The setting was an academic medical center Emergency Department serving primarily Latino and African-American patients. Change in self-reported pain intensity from baseline to 30 minutes postbaseline on a validated and reproducible 11-point numerical rating scale and count of adverse events were the primary outcomes. RESULTS: The sample consisted of 211 women and 144 men. The mean change in pain from baseline to 30 minutes postbaseline was 3.7 in women, 3.6 men (difference=0.04; 95% confidence interval: -0.52, 0.60). In women without nausea before administration of morphine, the incidence of adverse events was 18.3% versus 10.7% in men without initial nausea (difference=7.6%; 95% confidence interval: -2.0, 17.2). DISCUSSION: Men and women presenting to the Emergency Department did not have a differential response to a single weight-based dose of morphine for alleviation of acute pain. Women without baseline nausea had more adverse events than men. PMID- 18287824 TI - How common is back pain in women with gastrointestinal problems? AB - OBJECTIVE: This study examined the relationship between back pain and gastrointestinal (GI) symptoms in a large scale population study with consideration of possible confounding factors. METHODS: Cross-sectional analysis of survey data from the Australian Longitudinal Study on Women's Health was conducted using multinomial logistic regression to model 4 frequencies of back pain in relation to number of GI symptoms (including constipation, hemorrhoids, and other bowel problems). A total of 38,050 women from 3 age cohorts were included in analysis. RESULTS: After adjustment for confounding factors, the number of GI symptoms was significantly associated with back pain among all age cohorts. Odds ratios for experiencing back pain "rarely," "sometimes," and "often" increased with the number of GI symptoms. Young, mid-age, and older women who experience 2 or 3 GI symptoms had adjusted odds ratios of 3.3 (2.5 to 4.4), 3.0 (2.5 to 3.7) and 2.8 (2.3 to 3.4), respectively, for "often" having back pain. DISCUSSION: This study has identified a strong association between back pain and GI symptoms in women. Possible factors that may account for this relationship include referred pain through viscerosomatic convergence, altered pain perception, increased spinal loading when straining during defecation, or reduced support of the abdominal contents and spine secondary to changes in function of the abdominal muscles. PMID- 18287825 TI - Chronic pain conditions and suicidal ideation and suicide attempts: an epidemiologic perspective. AB - OBJECTIVES: Investigations of the association between chronic pain conditions and suicidal ideation (SI) and suicide attempts (SA) have rarely taken the effect of mental disorders into account and have been limited by nonrepresentative samples. The present study used a large population-based sample to investigate the association between chronic pain conditions and SI and SA. METHODS: Data were from the Canadian Community Health Survey Cycle 1.2 public use file conducted by Statistics Canada from 2001 to 2002 (N=36,984; response rate 77%). Respondents were asked if they had been diagnosed with the following painful conditions: migraine, back problems, arthritis, and fibromyalgia. Respondents were assessed for past 12-month SI and SA. The Composite International Diagnostic Interview was used to assess Diagnostic and Statistical Manual of Mental Disorders-IV. RESULTS: After adjusting for sociodemographics, Axis I mental disorders and comorbidity (3 or more mental disorders), the presence of 1 or more chronic pain conditions was associated with both SI and SA. Among respondents with a mental disorder, comorbidity with 1 or more chronic pain conditions was also associated with SI and SA. In models adjusting for other painful conditions, migraine had the strongest link with SI and SA. DISCUSSION: This is the first study to demonstrate the association between several chronic pain conditions and SI and SA while adjusting for mental disorders in a nationally representative sample. Moreover, this study demonstrates that among individuals with a mental disorder, having a chronic pain condition significantly increased the association with SI and SA. PMID- 18287826 TI - Acupuncture for treatment of persistent arm pain due to repetitive use: a randomized controlled clinical trial. AB - OBJECTIVE: To compare true and sham acupuncture in their abilities to relieve arm pain and improve arm function in individuals with arm pain due to repetitive use. METHODS: Participants with persistent arm pain (N=123) were randomly assigned to true or sham acupuncture groups and received 8 treatments over 4 weeks. The primary outcome was intensity of pain (10-point scale) and secondary outcomes were arm symptoms, arm function, and grip strength. Outcomes were measured during treatment (at 2 and 4 wk) and 1 month after treatment ended. RESULTS: Arm pain scores improved in both groups during the treatment period, but improvements were significantly greater in the sham group than in the true acupuncture group. This difference disappeared by 1 month after treatment ended. The true acupuncture group experienced more side effects, predominantly mild pain at time of treatments. DISCUSSION: Sham acupuncture reduced arm pain more than true acupuncture during treatment, but the difference did not persist after 1 month. Mild side effects from true acupuncture may have blunted any positive treatment effects. Overall, this study did not find evidence to support the effectiveness of true acupuncture in treatment of persistent arm pain due to repetitive use. PMID- 18287827 TI - The Brief Pain Inventory: pain's interference with functions is different in cancer pain compared with noncancer chronic pain. AB - OBJECTIVES: The Brief Pain Inventory (BPI) is a highly recommended and frequently used multidimensional pain assessment tool. The BPI includes 2 dimensions: pain intensity and pain's interference with functions. Our aims were to explore how patients respond to pain interference items by comparing responses from patients who had cancer with patients who had noncancer chronic pain (NCCP), and to explore how different levels of health-related quality of life affect upon pain's interference with functions. METHODS: Three hundred patients with cancer and 286 patients with NCCP were asked to complete the BPI and the European Organization for Research and Treatment of Cancer's Quality of Life Questionnaire (EORTC QLQ C30). The pain interference items were indexed into total interference, interference with physical functions, and interference with psychologic functions. Regression analyses were used to explore differences in pain's interference by group, pain intensity, and a possible interaction effect between them. The analyses were adjusted for age, sex, and all EORTC QLQ-C30 scales. RESULTS: The cancer patients reported higher values of physical interference than NCCP patients with the same level of pain intensity (P<0.001). NCCP patients reported higher values of psychologic interference than cancer patients (P=0.023). For total interference, these effects eliminated each other. When adjusting for age, sex, and the EORTC QLQ-C30 subscales, the results still remained significant except that adjusting for the subscale for physical function made the group effect insignificant for physical interference (P=0.30). DISCUSSION: The results indicate that patients are unable to report isolated pain's interference using the BPI. When reporting pain's interference with physical functioning, the level of physical functioning is more important than the level of pain. Patients' diagnoses have to be taken into account when interpreting reported pain's interference with functions. PMID- 18287828 TI - Carer experience of back pain is associated with adolescent back pain experience even when controlling for other carer and family factors. AB - OBJECTIVES: The study objectives were (1) to determine if adolescent back pain is related to carer back pain; and (2) to examine whether other carer and family factors accounted for any such relationship. METHODS: Back pain experience was collected independently from 1608 fourteen-year-old adolescents and their care givers by questionnaire. Measures of back pain impact on carers were also collected (modified activities, missed work, sought professional help, took medication). Other carer factors examined were age, sex, smoking habits, depression, anxiety, and stress. Familial factors examined included income, stressful events, and family functioning. Odds ratios were calculated to describe familial associations with multivariate modeling used to examine the influence of other carer and family factors. RESULTS: Lifetime prevalence of back pain was 46.5% for adolescents and 72% for their carers. Having a carer with back pain increased the risk of back pain for adolescents, and this was magnified if both carers had back pain (odds ratio 1.61). Adolescent back pain was not associated with primary carer age, sex, depression, anxiety, or stress but was related to family stressful events. However, these other factors did not diminish the association between carer back pain and adolescent back pain. DISCUSSION: Carer experience of back pain is clearly associated with adolescent experience of back pain. Although other carer and family factors are also associated, the carer's back pain experience is not a surrogate measure of these other factors suggesting other genetic and behavioral mechanisms may be important for adolescent back pain development. PMID- 18287829 TI - Pressure-pain threshold algometric measurement in patients with greater trochanteric pain after total hip arthroplasty. AB - BACKGROUND: The evaluation of tenderness associated with greater trochanteric pain (GTP) syndrome is amenable to bias and depends on the examiner's experience. In this study, we tested whether the use of an electronic pressure algometer enhanced the reliability of this evaluation. PATIENTS AND METHODS: Pressure-pain threshold (PPT) was measured with an electronic algometer in 18 patients who developed GTP after total hip arthroplasty and in matched controls. Both groups were evaluated with visual analog scale. RESULTS: The PPT measurements showed large interindividual variability across patients. The correspondence of the PPT measurements in asymptomatic patients was good. We found good validity for the algometer used. The PPT ratio of 0.8 (affected vs. unaffected side) can be used as a cut-off ratio. The PPT measurements at the greater trochanter (local pain) were significantly lower than at the ilio-tibial band (radiated pain). There was no correlation between PPT measurements and visual analog scales. Despite the acceptable sensitivity and specificity of pressure algometer, because of low positive predictive value and large interindividual variability, pressure algometer has a limited value as a screening test. CONCLUSIONS: The examination of tenderness associated with GTP is facilitated by the used algometer. It is the intraindividual body-side PPT differences that yield the most sensitive measurement for the assessment of deep pain. A cut-off value of 0.8 can be used for diagnostic purposes. Interindividual differences might be considerable and could mask pathologic diagnostic findings. PMID- 18287830 TI - Effects of rhythmic embrocation therapy with solum oil in chronic pain patients: a prospective observational study. AB - BACKGROUND: Chronic pain is one of the most frequent physical symptoms worldwide and requires intervention on many levels. Complementary therapies in particular are becoming increasingly relevant and popular in the treatment of chronic pain. The present study evaluates the therapeutic effects of rhythmic embrocation with a lavender/peat extract [Solum Oil (German: Solum Ol)]. MATERIALS AND METHODS: In a prospective observational study 100 patients with chronic pain were treated three times (T1-T3) within 24 days by rhythmic embrocation with Solum Oil. Main active constituents per 10 g of Solum Oil: aqueous peat extract (1:5) 1.96 g, and Lavandulae aetheroleum 0.5 g. The Mood Scale (Bf-S) and Pain Perception Scale (sensory PPS, affective PPS) were measured before the first and after each of the 3 treatments. To measure the therapeutic effects, effect sizes were calculated. RESULTS: Patients were mainly female (89%), mean duration of chronic pain (low back pain): 8.8 years; 1 drop-out due to urticaria. Bf-S was reduced from 25.8 [95% confidence interval (CI); 22.8-28.1] before T1 to 13.3 (95% CI; 11.7-15.0) after T3; sensory PPS from 18.8 (95% CI; 17.7-19.8) to 15.2 (95% CI; 14.1-16.4), affective PPS from 29.8 (95% CI; 27.9-31.7) to 21.3 (95% CI; 19.4-23.0) (all P<0.01). Effect sizes (d): Bf-S: d=0.81; affective PPs: d=0.85; sensory PPS: d=0.55. CONCLUSIONS: The high effect sizes indicate that repeated rhythmic embrocation with Solum Oil may improve mood, pain perception (sensory PPS), and the ability to cope with pain (affective PPS) in patients with chronic low back pain. To further assess the efficacy of this method, a randomized clinical trial is recommended. PMID- 18287831 TI - Rechargeable spinal cord stimulation versus non-rechargeable system for patients with failed back surgery syndrome: a cost-consequences analysis. AB - OBJECTIVES: Spinal cord stimulation (SCS) has been used for almost 40 years to treat refractory neuropathic pain after failed back surgery. Fully implantable non-rechargeable pulse generators have a battery life of between 2 and 5 years. A new SCS system with a rechargeable power source may last 10 to 25 years, or longer. The potential economic implications of longer battery life with a new SCS system has yet to be assessed. The study objective is to estimate the average difference in lifetime costs between rechargeable and non-rechargeable pulse generators used in treatment with SCS for failed back surgery syndrome. METHODS: A generalized state-transition probability framework was used to model costs. Input parameters for the base case analysis were obtained from several data sources including published literature, Medicare fee schedules, Medicare claims data, and expert opinion. RESULTS: A rechargeable SCS system is projected to require from 2.6 to 4.2 fewer battery generator replacements for battery depletion than a non-rechargeable SCS system. The total lifetime savings of a rechargeable system range from $104,000 to $168,833. In all of the one-way sensitivity analyses conducted, a rechargeable system saves money. Among all of the assumptions underlying the analysis, the annual cost after device removal contributes the most uncertainty. CONCLUSIONS: A rechargeable SCS system is projected to save up to $100,000 over a patient's lifetime. Fewer pulse generator replacements will also decrease patient discomfort and morbidity from procedural complications. PMID- 18287832 TI - Prevalence and characteristics of complaints of the arm, neck, and/or shoulder (CANS) in the open population. AB - OBJECTIVE: To study the prevalence of upper extremity disorders (UEDs) and neck as a total and complaints of the arm, neck and/or shoulder (CANS) not caused by acute trauma or any systemic disease as defined in the CANS model in the open population and to assess sociodemographic and health characteristics of chronic symptoms. METHODS: Data were obtained from the DMC3-study, a Dutch questionnaire survey on musculoskeletal conditions (>25 y, n=3664). Data on four anatomic sites were assessed: neck, shoulder, elbow, and wrist. Various health characteristics were measured including the Short Form-36. Rectangle diagrams were used to illustrate cooccurrence of pain in the four anatomic sites. RESULTS: The 12-month prevalence of CANS was 36.8%, the point prevalence was 26.4%, and 19.0% patients reported chronic CANS. Women, aged 45 to 64 years, with the lowest education level and working were the most affected. Within those with UEDs, around 25% of cases were caused by an acute trauma or by some systemic disease. Of those with chronic CANS, 58% reported use of healthcare. Healthcare users scored worse on general health, limitations in daily living, pain, and sickness absence than nonhealthcare users; >43% reported symptoms in more than 1 anatomic site. DISCUSSION: UEDs and CANS frequently occur in the open population. Excluding acute traumas and systemic diseases reduced the prevalence of CANS and resulted in a relatively healthier population. A compound definition of CANS seems indicated because of the large overlap of affected anatomic sites. PMID- 18287833 TI - Needle temperature effect on pain ratings after injection. AB - OBJECTIVES: Little research regarding appropriate immunization administration technique for adults has been carried out. Pain is a leading cause of self deferral from immunizations. The purpose of this study was to determine whether administering intramuscular injections using cold needles would decrease patients' perceived pain and have an effect on the immune response elicited by the vaccine. METHODS: Eighty participants received an injection of influenza vaccine in 1 arm and a saline injection in the other using a cold or room temperature needle in a double-blind fashion assigned at random. Participants rated their pain after each injection using a standard visual pain scale bounded by no pain and most painful injection ever. Vaccine antibody response was measured using hemagglutination inhibition assays. RESULTS: Overall, pain scores after influenza vaccine administration were quite low (34.2+/-2.5 mm). The mean pain score for influenza vaccine was not different using cold or room temperature needles (cold 32.2 mm+/-3.20 vs. room temperature 36.0 mm+/-3.80; t=0.76; P=0.450). The mean pain scores for saline injections did not differ (room temperature 23.7 mm+/-3.19 vs. cold 25.2+/-2.95; t=-0.34; P=0.73). Individuals receiving injections with cold needles had less bruising (0/40 vs. 5/40; P<0.02) at the injection site, but incidences of pain and erythema were similar. The use of cold needles for vaccine administration had no effect on antibody response. CONCLUSIONS: Pain after influenza vaccine administration is generally mild. Use of cold needles may not be worth pursuing for injections associated with mild pain. However, it may be worthwhile to explore using cold needles as an analgesic with more painful injections. PMID- 18287834 TI - Decision analysis for epidural labor analgesia with Multiattribute Utility (MAU) model. AB - OBJECTIVES: Epidural analgesia (EA) is an effective and safe method to relieve labor pain. Little is known about the factors associated with decision on EA. We applied Multiattribute Utility (MAU) theory to ascertain possible factors on which we based to predict whether parturients would receive EA or not (non-EA) during their labor. METHODS: A hierarchical questionnaire on the basis of MAU theory was designed by experts to include individual attributes, knowledge and attitude toward EA and cue factors. Items in the questionnaire were compared between the EA and the non-EA groups. Receiver operating characteristics curve was used to assess predictive validity of the MAU model. RESULTS: Of 167 parturients responding to the questionnaire, 151 participants (75 EA and 76 non EA groups) completed all questions. Parturients in the EA group had significantly higher education level (rate of junior college or above: 88% vs. 67%, P=0.002). There were also more primiparae in the EA group compared with non-EA group (76% vs. 46%, P<0.001). For items in MAU model, 12 out of 20 items revealed significant differences between the 2 groups. Among them, "fear of side effects," "fear of severe complications," and "fear of needle" had the most remarkable differences. The area under receiver operating characteristics equaled to 0.91 (95% confidence interval=0.86, 0.96) for pre-labor decision and 0.83 (95% confidence interval=0.76, 0.89) for final decision. DISCUSSION: These findings suggest that our MAU model can predict pre-labor decision and final decision of parturients by the incorporation of correlates with respect to knowledge and attitude. PMID- 18287835 TI - The role of TNF-alpha in patients with chronic low back pain-a prospective comparative longitudinal study. AB - OBJECTIVES: In this prospective longitudinal clinical study with a matched-pair design, we evaluated the role of tumor necrosis factor-alpha (TNF-alpha) and its clinical relevance in patients with chronic low back pain. METHODS: One hundred twenty patients with chronic low back pain were matched to a healthy control group. Patients and controls were prospectively followed for 6 months. At 4 fixed time points (day 0, day 10, day 20, and 6 mo) blood samples were taken and TNF alpha levels compared in the 2 groups, and correlations with pain level and clinical function were analyzed. RESULTS: At the beginning and at all other time points, there was a significantly higher proportion of TNF-alpha positive participants in the patients group than in the control group. The proportion of TNF-alpha positive patients decreased during the first 10 days of a multidisciplinary therapy in the patient group, but after this initial period, TNF-alpha levels remained constantly high with no further change until the final follow-up. In the healthy control group, the proportion of participants with positive TNF-alpha levels remained constant throughout the entire period. No significant correlation between TNF-alpha levels and pain or clinical function was detected. DISCUSSION: TNF-alpha seems to have a significant role in patients with chronic low back pain. However, the pathophysiology of this process, the clinical relevance of TNF-alpha and, especially, its part in a potential therapy regimen in these patients need to be more closely examined and defined in additional studies. PMID- 18287836 TI - Hepatobiliary scintigraphy in detecting lesser sac bile leak in postcholecystectomy patients: the need to recognize as a separate entity. AB - Cholecystectomy is one of the most commonly performed abdominal surgeries in which bile duct injury and bile leaks are the most important complications. Imaging plays an important role in the prompt diagnosis and management of bile leaks. The more common sites of bile leak are the gallbladder bed, subhepatic, in a bilioma, right paracolic gutter, or diffusely in the peritoneal cavity. Bile leak into the lesser sac (LS) is uncommon but is a special entity posing difficult problems in management. We have described in this study the clinical presentation, imaging findings, and management of 6 patients with biliary leakage into the LS postcholecystectomy. The clinical presentation of this condition was varied, ranging from patients with asymptomatic or with vague complaints resulting in difficulties in clinical suspicion or symptomatic but minimal enough not to be detected by ultrasonogram. Hepatobiliary scintigraphy played an important role in the diagnosis and management, and all patients required definitive therapeutic drainage procedures. Any persistent focal radiotracer activity in the anatomy of the LS, increasing with time and not diffusing into the general peritoneal cavity is diagnostic of bile leak into the LS. PMID- 18287837 TI - Fixed defect on rest/stress Tc-99m sestamibi study underestimates myocardial ischemia: comparison with 24-hour thallium-201 study for short- and intermediate term follow-up. AB - PURPOSE: We assessed whether a same day rest/stress gated Tc-99m sestamibi (MIBI) SPECT myocardial study underestimates reversible ischemia in patients with fixed perfusion defects compared with a 24-hour thallium-201 (Tl-201) study. The short- and intermediate-term outcome with or without Tl-201 reversibility was assessed. METHODS: Forty-nine consecutive patients with fixed MIBI defects received an additional Tl-201 study and were evaluated. Tl-201 was given to patients with a high clinical suspicion of underestimation of reversibility. Images were interpreted semiquantitatively by 3 nuclear medicine physicians using a 17 segment, 5-point model. A summed stress score (SSS) from stress MIBI images, a summed rest score (SRS) from Tl images, and a summed difference (SDS = SSS - SRS) score were calculated. SDS >3 indicated significant Tl-201 redistribution. Composite end points included acute myocardial infarction, unstable angina needing admission, cardiac death, or revascularization within 3 and 6 months. RESULTS: Fifteen of 49 patients showed no Tl-201 redistribution. Thirty-four of 49 (69%) patients had significant Tl-201 redistribution, and these patients had significantly higher cardiac events (CE) at 3 months (29% vs. 7%; P = 0.039), and higher at 6 months (32% vs. 7%; P = 0.027). These patients with CE had a larger amount of Tl-201 redistribution, mean SDS 8.6 vs. 5.3 (P = 0.047). Patients with significant Tl-201 redistribution had a lower left ventricular ejection fraction (mean 37%; P = 0.001). CONCLUSION: With short- and intermediate-term follow-up, our study shows a significant association towards fixed defects on the rest/stress MIBI study underestimating CE risk when compared with a delayed Tl 201 study, especially in patients with a large amount of Tl-201 redistribution. Hence, the addition of a Tl-201 study may be useful in the management of patients with large fixed MIBI defects, especially with a depressed left ventricular ejection fraction. PMID- 18287838 TI - FDG PET/CT findings in recurrent malignant schwannoma. AB - Malignant schwannoma is an uncommon but aggressive sarcoma that most commonly arises in young and middle-aged adults. We present a 28-year-old male patient with a recurrent chest wall malignant schwannoma. An FDG PET/CT was performed to evaluate the management of the patient. The PET/CT images showed hypermetabolic mass lesions in the left upper chest wall and increased FDG uptake in the left axillary region. The pathologic results confirmed the PET findings. We conclude that FDG PET/CT could play an important role in the staging, restaging, and post therapy follow-up of malignant schwannomas. PMID- 18287839 TI - FDG-PET on the trail of an unsuspected primary malignancy in the breast. AB - Proper identification of the primary malignancy can radically alter clinical management for the patient's benefit. This is a report of an unsuspected primary breast cancer in a patient being worked up for presumptive lymphoma. Prior investigation of lymphedema in the left lower extremity found widespread lymphadenopathy on computed tomography imaging, leading to initial biopsy revealing adenocarcinoma of unknown primary. F-18 fluorodeoxyglucose PET/computed tomography altered management by localizing an F-18 fluorodeoxyglucose avid breast nodule, directing breast biopsy with specific immunohistochemical analysis for breast cancer lineage in metastatic adenocarcinoma. The patient responded well to breast cancer-targeted chemotherapy. PMID- 18287840 TI - Cerebrospinal fluid leakage in intracranial hypotension syndrome: usefulness of indirect findings in radionuclide cisternography for detection and treatment monitoring. AB - PURPOSE: To evaluate indirect findings of cerebrospinal fluid (CSF) leakage on radionuclide cisternography and their changes after treatment. MATERIALS AND METHODS: This study was approved by the hospital's institutional review board and informed consent was obtained before each examination. A total of 67 patients who were clinically suspected of spontaneous intracranial hypotension (SIH) syndrome underwent radionuclide cisternography, and 27 patients who had direct findings of CSF leakage on radionuclide cisternography were selected for this evaluation. They were 16 males and 11 females, aged between 26 and 58 years. Sequential images of radionuclide cisternography were acquired at 1, 3, 5, and 24 hours after injection. We assessed the presence or absence of 4 indirect findings; early visualization of bladder activity, no visualization of activity over the brain convexities, rapid disappearance of spinal activity, and abnormal visualization of the root sleeves. Changes of the direct and indirect findings after treatment were also evaluated in 14 patients who underwent epidural blood patch treatment. RESULTS: Early visualization of bladder activity was found in all 27 patients. Seven of 27 (25.9%) patients showed no activity over the brain convexities. Rapid disappearance of spinal activity and abnormal root sleeve visualization were present in 2 (7.4%) and 5 (18.5%) patients, respectively. After epidural blood patch, both direct CSF leakage findings and indirect findings of early visualization of bladder activity had disappeared or improved in 12 of 14 patients (85.7%). The other indirect findings also disappeared after treatment in all cases. CONCLUSION: Indirect findings of radionuclide cisternography, especially early visualization of bladder activity, may be useful in the diagnosis and posttreatment follow-up of CSF leakage. PMID- 18287841 TI - Early detection of recurrent pediatric adrenal cortical carcinoma using FDG-PET. PMID- 18287842 TI - Intestinal duplication mimicking Meckel diverticulum. PMID- 18287843 TI - Increased FDG uptake in bilateral adrenal tuberculosis appearing like malignancy. PMID- 18287844 TI - Costal brown tumor detected by dual-phase parathyroid imaging and SPECT-CT in primary hyperparathyroidism. PMID- 18287845 TI - Tc-99m pertechnetate/sestamibi subtraction scan in a case of parathyroid carcinoma. PMID- 18287846 TI - Tc-99m sestamibi uptake mimicking parathyroid adenoma in a patient with primary hyperparathyroidism and occult multiple myeloma. PMID- 18287847 TI - Elucidation of acute renal failure due to recurrent non-Hodgkin lymphoma by F-18 FDG PET/CT. PMID- 18287848 TI - Primary splenic lymphoma detected by F-18 FDG PET. PMID- 18287849 TI - Methotrexate-induced and Epstein-Barr virus-associated B-cell lymphoma of the spine: MR and PET/CT imaging. PMID- 18287850 TI - F-18 FDG PET/CT assessment of gastrointestinal stromal tumor response to sunitinib malate therapy. PMID- 18287851 TI - Highlighting the role of FDG PET scan in early response assessment of gastrointestinal stromal tumor treated with imatinib mesylate. PMID- 18287852 TI - Apparent posterior misregistration of a liver lesion on PET/CT: pitfall secondary to partial hepatectomy. PMID- 18287853 TI - Uptake of Tc-99m methylene diphosphonate in peritoneal metastases from breast cancer. PMID- 18287854 TI - Imaging of systemic mastocytosis by FDG-PET/CT demonstrates increased activity in cortical bone. PMID- 18287855 TI - Detection of thyroid metastases from nasopharyngeal carcinoma with F-18 FDG PET/CT. PMID- 18287856 TI - Usefulness of lymphoscintigraphy in demonstrating lymphedema in patients with Noonan syndrome. PMID- 18287857 TI - I-123 metaiodobenzylguanidine (MIBG) in sympathetic denervation and reinnervation of salivary glands in a child with cervical neuroblastoma. PMID- 18287858 TI - Focal FDG uptake in the gastric contents: a PET/CT misregistration artifact due to vesicant use and water loading. PMID- 18287859 TI - F-18 FDG PET/CT demonstration of thymoma followed by development of lung cancer. AB - It is well-known that thymoma can be associated with diverse extrathymic malignancies. The authors present a case of a 72-year-old man with myasthenia gravis in which F-18 FDG PET/CT demonstrated high FDG uptake in both an anterior mediastinal mass and a lung nodule. The FDG uptake in the lung nodule was significantly higher than that of the thymic lesion, which might suggest the possibility of a second development of lung cancer in this thymoma patient. Surgical resection was done for the 2 tumors, and histologic examination revealed anterior mediastinal thymoma and primary adenocarcinoma of the lung. This case demonstrates the potential usefulness of PET/CT in detecting and assessing the second extrathymic malignancy in a patient with thymoma. PMID- 18287860 TI - Is it sufficient to quantify aspiration for predicting aspiration pneumonia? PMID- 18287862 TI - Trends in incidence of esophageal and gastric cardia cancer in high-risk areas in China. AB - Incidence rates have risen for gastric cardia adenocarcinoma, whereas rates have remained stable for esophageal squamous cell carcinoma in many western countries. The aim of this study was to describe and analyze trends in incidence rates for esophageal cancer and gastric cardia cancer in Cixian county, which is one of the high-risk areas for esophageal cancer in China as well as in the world. The data were obtained from the Cixian Cancer Registry, which is a population-based registry. All the data were checked and analyzed using SPSS 11.5. Between the years 1988 and 2003 there were 11,183 cases of esophageal cancer in the county. The age-standardized incidence rate was 168.55 per 100,000. In 1988, the age standardized incidence rate for male patients was 250.76/100,000; it declined to 160.05/100,000 in 2003, representing a decrease of 36.2%. The age-standardized incidence rate for female patients was 153.86/100,000 in 1988; it declined to 82.55/100,000 in 2003, showing a decrease of 46.3%. A slight decreasing trend was observed during the period, with a yearly decrease of 3.61% in male patients and 3.39% in female patients. For gastric cardia cancer, there were 1654 cases from 1988 to 2003. The age-standardized incidence rate was 25.58 per 100,000. The age standardized incidence rate for male patients was 13.75/100,000 in 1988; it increased to 28.55/100,000 in 2003, with a yearly increase of 7.65%. The age standardized incidence rate for female patients was 7.12/100,000 in 1988; it increased to 12.91/100,000, with a yearly increase of 5.44%. A statistically significant increasing trend of gastric cardia cancer was observed during the study period. Cixian county is a geographical region with a very high incidence of esophageal cancer and cardia cancer. The trend in the incidence rates of esophageal cancer had decreased slightly; on the other hand, gastric cardia cancer showed a significantly increased trend in the last 16 years. Detailed epidemiological analyses of demographic trends and risk factors will help to guide future cancer control strategies. PMID- 18287863 TI - Association of CYP1A1 Msp1 polymorphism with tobacco-related risk of gallbladder cancer in a north Indian population. AB - Polymorphisms in the gene encoding the metabolic enzyme cytochrome P450 1A1 (CYP1A1) might contribute to the variability in individual susceptibility to gallbladder cancer (GBC). This study comprised 142 consecutive cases of proven GBC and 171 healthy participants of similar ethnicity. CYP1A1 6235 T/C transition was analyzed using polymerase chain reaction-restriction fragment length polymorphism. The CC genotype of CYP1A1 was significantly associated with a risk of GBC [odds ratio (OR) 2.3, 95% confidence interval (CI)=1.1-4.5, P=0.026], and the age- and sex-adjusted OR was 2.0. The CYP1A1 C allele frequency was also higher in GBC patients (OR 1.4, 95% CI=1.0-2.0, P=0.039). After sex stratification, the TC genotype and C allele were significantly higher in men and conferred a risk for GBC (OR 4.2, 95% CI=1.8-9.7, P=0.001; OR 2.2, CI=1.3-3.7, P=0.005). In addition, the TC genotype and C allele frequencies were significantly higher in GBC patients without associated gallstones (OR 2.4, 95% CI=1.2-4.7, P=0.011; OR 1.6, CI=1.0-2.4, P=0.023). The usage of tobacco (smoking or nonsmoking) by GBC patients showed a significant increase in cancer risk with the TC genotype (OR 4.1, 95% CI=1.3-11.9, P=0.012). It seems that the higher risk of the TC genotype in men may be related to tobacco usage. PMID- 18287864 TI - The role of IGFBP3 functional polymorphisms in the risk of gastric cancer in a high-risk Chinese population. AB - Insulin-like growth factors (IGFs) and their receptors play a crucial role in regulating cell proliferation, differentiation, and apoptosis. Insulin-like growth factor-binding protein-3 is the most abundant insulin-like growth factor receptor in the serum and binds the majority of insulin-like growth factors. Studies reported that circulating level of insulin-like growth factor-binding protein-3 was modulated by functional genetic variants of insulin-like growth factor-binding protein-3 and, therefore, maybe associated with the risk of gastric cancer. In this case-control study of 576 gastric cancer cases and 647 cancer-free control participants in a high-risk Chinese population, we tested the hypothesis that functional polymorphisms A-202C and Gly32Ala of insulinlike growth factor-binding protein-3 are associated with risk of gastric cancer. We found that the variant 32Ala allele was associated with a significantly increased risk of gastric cancer (adjusted odds ratio=1.84, 95% confidence interval=1.45 2.33 for 32Gly/Ala and odds ratio=2.39, 95% confidence interval=1.47-3.90 for 32Ala/Ala, respectively), compared with the wild-type homozygote 32Gly/Gly. Although the A-202C variant was not significantly associated with gastric cancer risk in the single locus analysis, we found a significant locus-locus interaction between insulin-like growth factor-binding protein-3 A-202C and Gly32Ala loci on gastric cancer risk (Pint<0.001). These findings suggest that functional variants of insulin-like growth factor-binding protein-3 might be important markers for gastric cancer susceptibility and further studies are warranted to characterize the functional relevance of the locus-locus interaction of this gene. PMID- 18287865 TI - Nonsteroidal anti-inflammatory drug use and breast cancer risk: a Danish cohort study. AB - Epidemiologic studies investigating the effects of nonsteroidal anti-inflammatory drugs (NSAIDs) on breast cancer have yielded conflicting results. We examined the association between use of aspirin and nonaspirin NSAIDs and breast cancer risk among 28 695 women in the Danish Diet, Cancer and Health cohort. Information on NSAID and paracetamol use was obtained from a self-administered questionnaire completed at baseline (1993-1997) and updated through 2003 using a nationwide prescription database. Detailed information on breast cancer incidence and tumour characteristics was obtained from nationwide health registers. Cox proportional hazards regression was used to compute incidence rate ratios (RRs) and 95% confidence intervals (CIs). We identified 847 breast cancer cases over an average follow-up period of 7.5 years. Any NSAID use at baseline was associated with an increased incidence of breast cancer compared with nonuse (RR, 1.27; 95% CI, 1.10 1.45). A similar result was observed for any NSAID use in a combined analysis of baseline and prescription data (1.34; 95% CI, 1.15-1.56). Aspirin-only users experienced a slightly higher breast cancer incidence (RR, 1.38; 95% CI, 1.12 1.69) than exclusive users of nonaspirin NSAIDs (RR, 1.25; 95% CI, 1.04-1.49). Introduction of a lag time of 1 year provided similar results. We found no clear differences in risk estimates with frequency, recency or duration of NSAID use, or by hormone receptor status of the breast tumours. Paracetamol use was unrelated to breast cancer incidence. The increased breast cancer incidence among NSAID users may reflect a noncausal association, but our study provides no evidence of a chemopreventive effect of NSAIDs against breast cancer over the durations studied. PMID- 18287866 TI - Pesticides and prostate cancer: a review of epidemiologic studies with specific agricultural exposure information. AB - Prostate cancer is the most commonly diagnosed cancer in US men, and the second most commonly diagnosed cancer among men worldwide. Although pesticides have been implicated in studies of prostate cancer among farmers, meta-analyses have found heterogeneity across studies, and a number of exposures and lifestyle factors may be unique to farmers. The purpose of this paper is to review the epidemiologic literature to evaluate the hypothesis that agricultural exposure to pesticides is causally associated with prostate cancer risk. We analyzed the eight cohort studies and five case-control studies that quantified and/or evaluated agricultural exposure to particular pesticide classes or chemicals. Despite sporadic positive findings, these studies did not show consistently increased risks to support a causal association between agricultural pesticide use and prostate cancer. Studies using an 'external' comparison group must be interpreted in the context of confounding by differences in prostate-specific antigen screening intensity. Furthermore, most studies did not adjust for potential confounders other than age and time period. It is clearly not possible to exonerate any particular pesticide as a putative cause of prostate cancer - to do so would require an inverse empirical association with an upper confidence limit below the null value. Existing evidence does not point to any pesticide as satisfying widely used guidelines for establishing causation: a strong, exposure dependent and demonstrably unconfounded, unbiased association, documented in several studies. PMID- 18287867 TI - Case-control study of anaplastic thyroid cancer: goiter patients as controls. AB - Anaplastic thyroid cancer is very serious disease with bad prognosis and unknown etiology. The aim of the study was to test some hypotheses about other factors in addition to goiter related to anaplastic thyroid cancer occurrence. A case control study was performed during the period 1993-2005. The case group comprised 126 patients with newly diagnosed anaplastic thyroid cancer. The control group comprised 252 patients who had for the first time goiter operation, and had no malignancy of thyroid gland. Cases and controls were individually matched by age, sex and place of residence (urban/rural). According to conditional multivariate logistic regression analysis, anaplastic thyroid cancer was significantly related to lower education (odds ratio=1.85, 95% confidence interval=1.21-2.82), other malignant tumors in personal history (odds ratio=4.37, 95% confidence interval=1.11-17.31), blood group B (odds ratio=3.69, 95% confidence interval=1.10-12.49), menarche at >or=15 years of age (odds ratio=2.63, 95% confidence interval=1.15-5.88), and first full-term pregnancy before 19 years of age (odds ratio=2.96, 95% confidence interval=1.26-6.96). On the basis of the results obtained, risk factors for anaplastic thyroid cancer are similar to risk factors for differentiated thyroid cancers. PMID- 18287868 TI - Alcohol and laryngeal cancer: an update. AB - Epidemiological studies provide definite evidence that alcohol drinking is an independent risk factor for laryngeal cancer. This risk increases with the amount of alcohol consumed: in a meta-analysis of 20 studies conducted in North America, Europe, Japan and Korea the multivariate relative risks were about 2 for 50 g ( approximately 4 drinks)/day and about 4 for 100 g/day compared to nondrinkers, in the absence of evidence of a threshold. Genetic polymorphisms in the alcohol metabolizing enzyme aldehyde dehydrogenases have been found to be associated with upper aerodigestive tract cancer, including the larynx. Further, the risk increases by concomitant tobacco smoking, each agent approximately multiplying the effect of the other. In the absence of smoking, the relative and absolute risks are small for moderate alcohol consumption, but there is an increased risk for elevated alcohol consumption. After stopping drinking, some fall in risk becomes apparent only in the long term. The supraglottis is more closely related to alcohol consumption, as compared to the glottis/subglottis. In various populations, the most commonly used alcoholic beverage appears to be the one most strongly associated with laryngeal cancer risk, suggesting that no meaningful difference exists for different types of alcoholic beverages. PMID- 18287869 TI - Association of cytochrome P450, glutathione S-transferase and N-acetyl transferase 2 gene polymorphisms with incidence of acute myeloid leukemia. AB - The objective of the paper was to study the association of polymorphisms of phases I and II xenobiotic metabolizing enzyme genes cytochrome P450 (CYP 4501A1*2A, *2B, *2C and *4 alleles, CYP-4502D6*4 allele), glutathione-S transferase (GSTM1 and GSTT1 null genotypes) and N-acetyl transferase 2 (NAT2*6B and *7A alleles) with the incidence of acute myeloid leukemia (AML) in an eastern Indian population. Polymerase chain reaction and restriction fragment length polymorphism of genomic DNA from peripheral blood cells were used to detect CYP 450 and NAT2 gene polymorphisms in 110 AML patients and 144 racially and geographically matched normal controls. Polymerase chain reaction was also applied to detect GST gene polymorphisms in both groups. A statistically significant difference between the AML group and the normal group was observed in the case of glutathione-S-transferase M1 null (odds ratio 3.25, 95% confidence interval 1.9-5.58, P<0.001) and N-acetyl transferase 2*6B (odds ratio 3.04, 95% confidence interval 1.79-5.16, P<0.001) genotypes. Combined deficiency of N acetyl transferase 2 and glutathione-S-transferase M1 genes produced an odds ratio of 11.91 (95% confidence interval 4.06-34.96, P<0.001). The effect of N acetyl transferase 2*6B (P<0.001) is significant only at ages