Immunohistochemical detection of changes in growth hormone cells in rat pituitaries in protein deficiency.
1. An immunohistochemical method was used to study the effect of a low-protein diet on growth hormone (GH) cells in the pituitaries of developing rats. The deficient diet (80 g protein/kg) was administered during gestation and lactation, or during the time after weaning until 90 d of age, or during both periods. 2. GH-cell changes were much more striking in males than in females. 3. In males, GH-producing cells were usually reduced in size and number in all treatments. The effect was most intense when protein deprivation occurred throughout gestation and sucking, and continued until 90 d of age, but it was also evident in animals given the low-protein diet only after weaning. Recuperation appeared to be almost complete when offspring of deprived dams were fed on a normal diet after weaning. 4. It is concluded that a low-protein diet reduces the amount of GH in the rat pituitary in a way similar to that with a protein-free diet.


Validation of eGFP fluorescence intensity for testing in vitro cytotoxicity according to ISO 10993-5.
ISO 10993-5 provides one of the accepted standards for testing the biotoxicity of new materials. All of the recommended test procedures rely upon the uptake or metabolism of dye by living cells. Results of direct contact tests can be potentially compromised by interaction or adsorption of the dye or its metabolic products. Therefore, the aim of the current study was to validate the use of the eGFP signal of transfected NIH-3T3 fibroblasts with the results of the MTT test in order to provide a test procedure that is very close to the ISO 10993-5 but has the advantage of not relying on the addition of dye. Our tests show that the MTT assay detects cytotoxicity in the eGFP NIH-3T3 cells at least as well as in the L929 cells. To facilitate the validation, we chose to integrate the fluorescence measurements into the MTT test procedure. To that end, an additional washing step was introduced. Additionally, medium without phenol red was used, resulting in a very high correlation of both measurements. Without these modifications, the fluorescence test was comparable to the MTT test in its ability to detect the cytotoxic potential of substances; however, it did result in slightly elevated IC50 concentrations. As the results of both tests correlated highly, measurement of the eGFP signal appears to present a reliable tool for detecting cytotoxicity of materials in line with the ISO 10993-5 norm with the advantage of avoiding the addition of dyes and the subsequent potential interaction with test materials. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 715-722, 2017.


Differentiation of antemortem pulmonary thromboembolism and postmortem clot with unenhanced MRI: a case report.
We report a case of a 39-year-old woman who died of fulminant pulmonary thromboembolism (PE). Autopsy showed classical findings of fulminant PE with occlusion of the bilateral main stem pulmonary arteries. Ancillary testing revealed inherited thrombophilia (Prothrombin 20,210 G > A and MTHFR 677 C > T mutation). Pre-autopsy postmortem computed tomography was used to test whether virtual imaging studies alone (virtual autopsy) would be sufficient to detect PE. Our studies show that computed tomography (CT) can differentiate antemortem clots from a postmortem clot in certain cases, particularly when combined with magnetic resonance imaging (MRI), which is superior in the assessment of soft tissue. We show that postmortem CT and MRI can aid in the diagnosis of pulmonary embolism by virtual autopsy, especially when used in conjunction.


Myocardial ischemia and reperfusion: the role of oxygen radicals in tissue injury.
Thrombolytic therapy has gained widespread acceptance as a means of treating coronary artery thrombosis in patients with acute myocardial infarction. Although experimental data have demonstrated that timely reperfusion limits the extent of infarction caused by regional ischemia, there is growing evidence that reperfusion is associated with an inflammatory response to ischemia that exacerbates the tissue injury. Ischemic myocardium releases archidonate and complement-derived chemotactic factors, e.g., leukotriene B4 and C5a, which attract and activate neutrophils. Reperfusion of ischemic myocardium accelerates the influx of neutrophils, which release reactive oxygen products, such as superoxide anion and hydrogen peroxide, resulting in the formation of a hydroxyl radical and hypochlorous acid. The latter two species may damage viable endothelial cells and myocytes via the peroxidation of lipids and oxidation of protein sulfhydryl groups, leading to perturbations of membrane permeability and enzyme function. Neutrophil depletion by antiserum and inhibition of neutrophil function by drugs, e.g., ibuprofen, prostaglandins (prostacyclin and PGE1), or a monoclonal antibody, to the adherence-promoting glycoprotein Mo-1 receptor, have been shown to limit the extent of canine myocardial injury due to coronary artery occlusion/reperfusion. Recent studies have challenged the hypothesis that xanthine-oxidase-derived oxygen radicals are a cause of reperfusion injury. Treatment with allopurinol or oxypurinol may exert beneficial effects on ischemic myocardium that are unrelated to the inhibition of xanthine oxidase. Furthermore, the human heart may lack xanthine oxidase activity. Further basic research is needed, therefore, to clarify the importance of xanthine oxidase in the pathophysiology of reperfusion injury.(ABSTRACT TRUNCATED AT 250 WORDS)


Roles of transferrin receptors in erythropoiesis.
Erythropoiesis requires large amounts of iron for hemoglobin synthesis, which is mainly provided by macrophages and the intestines in a transferrin (Tf)-bound form. Bone marrow erythroblasts incorporate Tf through endocytosis, which is mediated by transferrin receptor 1 (TFR1). Recently, human TFR1, aside from its role as a Tf receptor, was also found to be a receptor for the H-subunit of ferritin (FTH). In humans, hematopoietic erythroid precursor cells express high levels of TFR1 and specifically take up the FTH homopolymer (H-ferritin). H-ferritin inhibits the formation of burst forming unit-erythroid colonies in vitro. TFR2, which is also a Tf receptor, is predominantly expressed in hepatocytes and erythroid precursor cells. In the liver, TFR2 forms a complex with HFE, a hereditary hemochromatosis-associated protein, and acts as an iron sensor. In mice, hepatocyte-specific knockout of the TFR2 gene has been shown to cause systemic iron-overload with decreased expression of hepcidin, the central regulator of iron homeostasis. In erythroid cells, TFR2 forms a complex with the erythropoietin receptor and facilitates its trafficking to the cell membrane. Moreover, hematopoietic cell-specific knockout of the TFR2 gene causes microcytic erythrocytosis in mice. This review focuses on the molecular evolution and functions of these TFRs and their ligands.


Microcomplex--a new incomplete heart block pattern.
This article reports a new, transient, but identical electrocardiogram presentation in 2 patients at 2 different institutions. While its mechanism is uncertain, analysis by exclusion suggests that a diffusely prolonged but nonhomogenous myocardial refractoriness may have altered both QRS morphology and amplitude and also may have resulted in periods of 2:1 AV block.


[Intraocular irrigating solutions during phacoemulsification and corneal changes].
This study evaluate the number, density and quality of endothelial cells of the cornea during phacoemulsification when several sterile intraocular irrigating solutions are used.


alpha-Tocopherol, an inhibitor of epidermal lipid peroxidation, prevents ultraviolet radiation from suppressing the skin immune system.
We investigated the involvement of epidermal lipid peroxidation in the induction of ultraviolet radiation (UVR)-induced suppression of the skin immune system. The shaved dorsal skin of C3H/HeJ mice was irradiated with one of two subinflammatory solar-simulated UVR protocols 3 days per week for 4 weeks. Then half of 1 mg, 1, 2.5 or 5 mg alpha-tocopherol in a vehicle of acetone was topically applied to the shaved dorsal skin before UVR, A 5 mg dose of vitamin E gave complete protection against a UVR protocol that induced a 55% reduction in the contact hypersensitivity response to 2,4,6-trinitrochlorobenzene and a 23% reduction in epidermal Langerhans cell density. Lower doses were ineffective. alpha-Tocopherol was unable to protect against a higher UVR protocol. As 5 mg alpha-tocopherol did not prevent postirradiation inflammatory edema it is unlikely that the antioxidant acted as a sunscreen. However, 5 mg alpha-tocopherol inhibited UVR-induced epidermal lipid peroxidation, suggesting that this may be one mechanism by which alpha-tocopherol prevented UVR-induced local immunosuppression. Scavenging of UVR-generated lipid peroxides and reactive oxygen may have inhibited loss of cell membrane integrity preventing depletion of LC numbers, thus protecting from local immunosuppression.


Analysis of the long-term results of living donor liver transplantation in adults.
Living donor liver transplantation (LDLT) is an alternative to conventional transplantation given its excellent results. The aim of this study is to evaluate long-term outcomes in LDLT recipients. 100 consecutive THDV recipients from the Hospital Clínic of Barcelona from March 2000 to October 2015 were included. The main indication for transplantation was end-stage liver disease (58%) followed by hepatocellular carcinoma (41%). 95% of grafts consisted of the right liver of the donor and the 5% of the left liver. After a median follow-up of 65.5 months, patient and graft survival at 1, 3, and 5 years was 93%, 80% and 74% and 90%, 76%, and 71%, respectively. The overall re-transplant rate was 9%. The most common long-term complication was biliary stenosis (40%) with an average time of onset of 13.5±12 months, with repeated admissions and an average of 1.9±2 endoscopic procedures and 3.5±3 Radiological procedures per patient. The definitive treatment was radiological dilation in 40% of cases, surgical intervention in 22.5% and re-transplantation in 7.5%. Given the long-term results, LDLT is confirmed as an alternative to conventional transplantation. However, the high rate of late biliary complications involves repeated admissions and invasive treatments that, while not compromising survival, can affect the patient's quality of life.


A green approach to offset the perturbation action of 1-butyl-3-methylimidazolium iodide on α-chymotrypsin.
Recently, studies have provided evidence for the negative effects of ionic liquids (ILs), "greener solvents," on proteins [Phys. Chem. Chem. Phys., 2014, 16, 5514]. The search to offset the negative effects of ILs on proteins has come into limelight as the maintenance of a "green solvent medium" is a great challenge for chemists and biochemists. As the first step in this search, 1-butyl-3-methylimidazolium bromide ([Bmim][Br]) has been applied to offset the deleterious action of 1-butyl-3-methylimidazolium iodide ([Bmim][I]) on α-chymotrypsin (CT). Fluorescence and circular dichroism (CD) experiments results have indicated that [Bmim][Br] significantly offsets the deleterious action of [Bmim][I] at lower concentrations (0.025 M). Surprisingly, the stabilizing action of [Bmim][Br] turns into a deleterious action for CT at higher concentrations (>0.1 M). On the other hand, [Bmim][I] acted as a destabilizer for CT at all investigated concentrations (0.025-0.6 M). The results obtained from this study lead to valuable insights into the selection of suitable ILs to attenuate the deleterious action of another IL without disturbing the protein structure.


Nutritional support in acute pancreatitis.
This review explores the role of enteral and parenteral nutrition in severe acute pancreatitis and discusses the potential benefits of glutamine, omega-3 fatty acids, arginine and selenium together with probiotics and prebiotics in these patients. In addition, the method of refeeding during the convalescent period is also examined. A complex picture is emerging in which enteral nutritional support may be important early in the course of the disease with parenteral nutrition being used more as a backup and possibly only after the systemic inflammatory response has peaked. Nasogastric feeding, sometimes supplemented by parenteral nutrition, is as efficacious as nasojejunal feeding. An individualized approach, in which strategies of nutritional support are tailored to patient response, is gaining currency. Data regarding specialized formulae are mixed but the use of prebiotics is showing promise and is worthy of further exploration. In the convalescent period, preliminary data also indicate that the risk of pain developing is no greater if a light diet is instituted rather than clear fluids. Nutritional support in acute pancreatitis remains challenging and controversial with a number of different and unexpected approaches, including the use of nasogastric feeding and dual enteral and parenteral nutrition support, being adopted in recent clinical trials.


Concentrations of trace elements in tissues of red fox (Vulpes vulpes) and stone marten (Martes foina) from suburban and rural areas in Croatia.
Trace elements concentrations (As, Cd, Cu, Pb and Hg) were determined in the liver, kidney and muscle of 28 red fox (Vulpes vulpes) and 16 stone marten (Martes foina) from suburban and rural habitats from Croatia. Rural and suburban habitats affected Cd and Hg levels in the muscle, liver and kidney of red fox. Significant differences in metal concentrations in the muscle, liver and kidney were detected among species. Suburban stone marten accumulated the highest levels of trace elements (mg/kg w.w.): in muscle 0.019 for Hg; in liver 0.161 for Cd, 36.1 for Cu and 0.349 for Pb; in kidney 1.34 for Cd and 0.318 for Pb. Values observed were higher than those found in suburban red fox and therefore, may represent an important bioindicator for the accumulation of toxic metals in urbanized habitats.


Coupling between Chemical and Meteorological Processes under Persistent Cold-Air Pool Conditions: Evolution of Wintertime PM2.5 Pollution Events and N2O5 Observations in Utah's Salt Lake Valley.
The Salt Lake Valley experiences severe fine particulate matter pollution episodes in winter during persistent cold-air pools (PCAPs). We employ measurements throughout an entire winter from different elevations to examine the chemical and dynamical processes driving these episodes. Whereas primary pollutants such as NOx and CO were enhanced twofold during PCAPs, O3 concentrations were approximately threefold lower. Atmospheric composition varies strongly with altitude within a PCAP at night with lower NOx and higher oxidants (O3) and oxidized reactive nitrogen (N2O5) aloft. We present observations of N2O5 during PCAPs that provide evidence for its role in cold-pool nitrate formation. Our observations suggest that nighttime and early morning chemistry in the upper levels of a PCAP plays an important role in aerosol nitrate formation. Subsequent daytime mixing enhances surface PM2.5 by dispersing the aerosol throughout the PCAP. As pollutants accumulate and deplete oxidants, nitrate chemistry becomes less active during the later stages of the pollution episodes. This leads to distinct stages of PM2.5 pollution episodes, starting with a period of PM2.5 buildup and followed by a period with plateauing concentrations. We discuss the implications of these findings for mitigation strategies.


The influence of age on the response of major depression to electroconvulsive therapy: a C.O.R.E. Report.
As part of a C.O.R.E., multi-site longitudinal study comparing continuation electroconvulsive therapy (ECT) vs. continuation pharmacotherapy, the authors determined the response of 253 patients with major depression to acute-phase, bilateral ECT by use of the 24-item Hamilton Rating Scale for Depression. Remission rates for three age-groups, > or =65 years; 46-64 years; and < or =45 years, were 90 percent, 89.8 percent, and 70 percent, respectively. Age, as a continuous variable, positively influenced response to treatment. Bilateral, dose-titrated ECT is a highly effective acute treatment for major depression, and older age confers a greater likelihood of achieving remission.


Comparison of steady-state serum concentrations of digoxin in tablets (Lanoxin) and capsules (Lanoxicaps) in the elderly.
The bioavailability of digoxin in solution (Lanoxicaps, 90-100 percent) is superior to that of Lanoxin tablets (60-80 percent) in young healthy volunteers. The alleged bioequivalence of digoxin in tablets (Lanoxin 0.25, 0.125 mg) and capsules (Lanoxicaps 0.2, 0.1 mg) was studied in 20 elderly inpatients (14 women, 6 men, aged 84 +/- 5 years), treated with digoxin for atrial fibrillation. In a crossover design, steady-state digoxin concentrations were measured in 16 patients after once-daily administration of Lanoxin 0.125 mg or Lanoxicaps 0.1 mg, each for at least ten days. Four other patients took Lanoxin 0.25 mg and Lanoxicaps 0.2 mg successively. The steady-state digoxin concentrations were statistically significantly different, being 1.3 +/- 0.5 nmol/L for Lanoxin and 0.9 +/- 0.4 nmol/L for Lanoxicaps (p = 0.003). Whereas in young healthy volunteers the bioavailability of digoxin from Lanoxicaps (90-100 percent) exceeds that from Lanoxin (80 percent), in elderly patients the absorption appears to be similar. This suggests a lower than expected bioavailability of Lanoxicaps, which may result in subtherapeutic serum concentrations.


A systematic review and economic evaluation of epoetin alpha, epoetin beta and darbepoetin alpha in anaemia associated with cancer, especially that attributable to cancer treatment.
To assess the effectiveness and cost-effectiveness of epoetin alpha, epoetin beta and darbepoetin alpha (referred to collectively in this report as epo) in anaemia associated with cancer, especially that attributable to cancer treatment. Electronic databases were searched from 2000 (1996 in the case of darbepoetin alpha) to September 2004. Using a recently published Cochrane review as the starting point, a systematic review of recent randomised controlled trials (RCTs) comparing epo with best standard was conducted. Inclusion, quality assessment and data abstraction were undertaken in duplicate. Where possible, meta-analysis was employed. The economic assessment consisted of a systematic review of past economic evaluations, an assessment of economic models submitted by the manufacturers of the three epo agents and development of a new individual sampling model (the Birmingham epo model). In total 46 RCTs were included within this systematic review, 27 of which had been included in the Cochrane systematic review. All 46 trials compared epo plus supportive care for anaemia (including transfusions), with supportive care for anaemia (including transfusions), alone. Haematological response (defined as an improvement by 2 g/dl(-1)) had a relative risk of 3.4 [95% confidence interval (CI) 3.0 to 3.8, 22 RCTs] with a response rate for epo of 53%. The trial duration was most commonly 16-20 weeks. There was little statistical heterogeneity in the estimate of haematological response, and there were no important differences between the subgroups examined. Haemoglobin (Hb) change showed a weighted mean difference of 1.63 g/dl(-1) (95% CI 1.46 to 1.80) in favour of epo. Treatment with erythropoietin in patients with cancer-induced anaemia reduces the number of patients who receive a red blood cell transfusion (RBCT) by an estimated 18%. Health-related quality of life (HRQoL) data were analysed using vote counting and qualitative assessment and a positive effect was observed in favour of an improved HRQoL for patients on epo. Published information on side-effects was of poor quality. New trials provided further evidence of side-effects with epo, particularly thrombic events, but it is still unclear whether these could be accounted for by chance alone. The results of the previous Cochrane review had suggested a survival advantage for epo (HR 0.84, 95% CI 0.69 to 1.02), based on 19 RCTs. The update, based on 28 RCTs, suggests no difference (HR 1.03, 95% CI 0.88 to 1.21). Subgroup analysis suggested some explanations for this heterogeneity, but it is difficult to draw firm conclusions without access to the substantial amounts of missing or unpublished data, or more detailed results from some of the trials with heterogeneous patient populations. The conclusions are, however, broadly in line with those of a Food and Drug Administration (FDA) safety briefing, which recommended that patients with a haemoglobin above 12 g/dl(-1) should not be treated; the target rate of rise in Hb should not be too great, and further carefully conducted trials are required to determine which subgroups of patients may be harmed by the use of these products, in particular through the stimulation of tumour activity. Five published economic evaluations identified from the literature had inconsistent results, with estimates ranging from a cost per quality-adjusted life-year (QALY) under pound 10,000 through to epo being less effective and more costly than standard care. The more favourable evaluations assumed a survival advantage for epo. The three company models submitted each relied on assumed survival gains to achieve relatively low cost per QALY, from pound 13,000 to pound 28,000, but generated estimates from pound 84,000 to pound 159,000 per QALY when no survival gain was assumed. Each of these models relied on Hb levels alone driving utility, and each assumed gradual normalisation of Hb in the standard treatment arm after the end of treatment. The Birmingham epo model followed the company models in regard to the relationship between Hb levels and utility, and also assumed normalisation in the base case. With no survival gain, the incremental cost per QALY was pound 150,000, falling to pound 40,000 when the lower, more favourable, confidence interval for survival was used. Epo is effective in improving haematological response and reducing RBCT requirements, and appears to have a positive effect on HRQoL. The incidence of side-effects and effects on survival remains highly uncertain. However, if there is no impact on survival, it seems highly unlikely that epo would be considered a cost-effective use of healthcare resources. The main target for further research should be improving estimates of impact on survival, initially through more detailed secondary research, such as the individual patient data meta-analysis started by the Cochrane group. Further trials may be required, and have been recommended by the FDA, although many trials are in progress, completed but unreported or awaiting mature follow-up. The Birmingham epo model developed as part of this project contains new features that improve its flexibility in exploring different scenarios; further refinement and validation would therefore be of assistance. Finally, further research to resolve uncertainty about other parameters, particularly quality of life, adverse events, and the rate of normalisation, would also be beneficial.


Dietary lipids modulate the expression of miR-107, a miRNA that regulates the circadian system.
The increased prevalence of cardiovascular diseases (CVDs) has been hypothesized to be the result of an increased exposure to a host of atherogenic environmental factors, paramount among them being unhealthy dietary habits. Long-chain n-3 polyunsaturated fatty acids (PUFAs) have been shown to have cardio protective effects, partially due to their ability to regulate gene expression. In this regard, increasing attention has been devoted to the role of miRNAs as regulators of multiple metabolic pathways whose deregulation has been associated with CVD risk. In this work we investigated whether miRNA expression was regulated by docosahexanoic acid, conjugated linoleic acid and cholesterol in Caco-2 cells. Among the modulated miRNAs, miR-107 was differentially expressed by all treatments and this modulation was independent of its hosting gene, panthothenate kinase 1, possibly through its own promoter, which contains binding sites for metabolically relevant transcription factors. Among the putative target genes of miR-107, we found some genes with key roles in circadian rhythm. Specifically, we demonstrated that binding of miR-107 to the circadian locomotor output cycles kaput gene results in the deregulation of the circadian rhythm of the cells. Since chronodisruption has been linked to metabolic disorders such as type 2 diabetes, atherosclerosis, obesity, and CVD, our findings suggest that miR-107 could represent a new approach for pharmacological treatment of these diseases.


Agrimonia eupatoria L. (Agrimony) Extract Alters Liver Health in Subjects with Elevated Alanine Transaminase Levels: A Controlled, Randomized, and Double-Blind Trial.
Agrimonia eupatoria L. has been shown to protect against liver injury due to its lipid lowering and antioxidant activities. The aim of this research was to evaluate the effect of A. eupatoria L. aqueous extract (AEE) on 80 subjects with elevated alanine transaminase (ALT) levels in a randomized, double-blind, placebo-controlled, 8-week study. This trial was conducted between January 2013 and July 2013 at the Oriental Medical Hospital (Jecheon) of Semyung University. The trial included subjects aged 20 years or older who were diagnosed with mildly to moderately elevated ALT levels (between 45 and 135 IU/L). Subjects received two capsules of placebo or AEE twice a day for 8 weeks. Adverse events were recorded. Eighty subjects were randomized to placebo or AEE groups who had similar baseline characteristics. During the 8 weeks of treatment, 11 subjects were excluded from the analysis for protocol violation or consent withdrawal; efficacy of treatment was, therefore, evaluated in 69 subjects (placebo = 35, AEE = 34). The AEE group showed a significant reduction in ALT and serum triglyceride (TG) at 8 weeks compared with the placebo group (ALT P = .044, TG P = .020). Significant group and time interactions were found in ALT (P = .038), aspartate aminotransferase (P = .040), and TG (P = .010). Alkaline phosphatase, total bilirubin, and gamma-glutamyl transferase levels were not different between the two groups. There were no reported severe adverse events during this study, and total protein, albumin, blood urea nitrogen, creatine, and total cholesterol levels were normal in both groups. AEE consumption was safe and generally well tolerated without severe adverse events.


Appearance of early and late components of Epstein-Barr virus-associated membrane antigen in Daudi cells superinfected with P3HR-1 virus.
This study investigated the synthesis of membrane antigen (MA) as well as virus capsid antigen (VCA) and early antigen (EA) in Daudi cells which had been superinfected with the P3HR-1 strain of Epstein-Barr virus (EBV) and then treated with trypsin to remove initially absorbed MA-positive material from the cell surface. Synthesis of MA, VCA and EA was completely inhibited by puromycin. A marked reduction in the frequency of MA positive cells was observed in superinfected cells cultured in the presence of either cytosine arabinoside (Ara-C) or phosphonoacetate (PA); however, a small fraction of MA synthesis occurred, suggesting an inhibitor insensitive component in MA, A differential absorption of EBV antibody-positive human serum with the Ara-C treated or untreated infected cells detected two antigenically different components in MA: early (Ara-C insensitive) and late (Ara-C sensitive) MA.


Opioid prescribing patterns in emergency departments and future opioid use in adolescent patients.
Evidence suggests that exposure to opioids in adolescence increases risk of future opioid use. We evaluate if exposure to high versus low intensity opioid prescribers in the Emergency Department (ED) influences the risk of future opioid use in adolescents. Retrospective study of opioid-naïve patients 10 to 17 years seen in one of 14 EDs between January 2013 and December 2014. We categorized ED providers into quartiles according to the proportion of encounters resulting in opioid prescriptions. Primary outcome was use of opioids in the subsequent 12 months. Analysis adjusted for patient characteristics and compared future use of opioids for patients seen by the lowest versus the highest prescribing quartiles. We included 9,688 patient encounters evaluated by the lowest opioid prescribing physician quartile versus 9,467 in the highest. The highest quartile gave opioid prescriptions to 14.9% of their patients compared to 2.8% for the lowest quartile. No association with future opioid use was found for patients evaluated by low versus high prescriber quartiles (OR 0.99, 95% CI 0.90-1.08). Patients with increasing age (OR 2.15, 95% CI 1.92-2.42) and white versus Hispanic ethnicity (OR 1.55, 95% CI 1.33-1.80) were associated with recurrent opioid use. We found no association between high intensity opioid prescribers and recurrent 12 month use of opioids in opioid-naïve adolescents seen in the ED. This likely reflects various factors that put adolescents at risk for recurrent opioid use and may indicate the importance of the second prescription from primary care after initial exposure to opioids.


Neonatal bronchopulmonary dysplasia predicts abnormal pulmonary HRCT scans in long-term survivors of extreme preterm birth.
There is an increasing understanding that extreme preterm birth carries a risk of long-term pulmonary sequelae. A study was undertaken to investigate if, and in what way, neonatal factors were associated with subsequent abnormalities on pulmonary high-resolution CT (HRCT) scanning and if pulmonary function was related to these abnormalities. HRCT scanning and pulmonary function tests were performed less than 2 weeks apart in 74/86 eligible subjects (86%) born at a gestational age of < or =28 weeks or with a birth weight of < or =1000 g within a defined area in Western Norway in 1982-5 (n = 42) or 1991-2 (n = 32). Mean age at examination was 18 and 10 years, respectively. HRCT scans were interpreted by a paediatric radiologist blinded to the clinical data using a structured system allowing scores from 0 to 50. Lung parenchymal abnormalities were found in 64 subjects (86%), the median (interquartile range) score being 3.0 (1.75-5.0) points. Prolonged neonatal requirement for oxygen treatment predicted poor outcome, and an increase of 100 days increased the average HRCT score by 3.8 points (p<0.001). There was also a positive association of the severity of pulmonary function abnormalities with the extent of HRCT abnormalities, exemplified by the relation between forced expiratory volume in 1 s and total HRCT score (beta = -0.090; p<0.001). In area-based cohorts of long-term survivors of extremely preterm birth, prolonged neonatal requirements for oxygen treatment predicted subsequent structural abnormalities on HRCT scans and in pulmonary function, and these two outcome measures were interrelated.


The unusual suspects--innate lymphoid cells as novel therapeutic targets in IBD.
Innate lymphoid cells (ILCs) are a family of immune cells that selectively accumulate in mucosal tissues serving as sentinels at the vanguard of host protective immunity. However, they are also implicated as cellular mediators of immune-mediated diseases, most notably IBD. ILCs are subdivided into distinct lineages based on the expression of effector cytokines and master transcription factors that programme their differentiation and inflammatory behaviour. Strikingly, these subsets closely resemble CD4(+) T-cell lineages, including T helper (TH)1, TH2 and TH17 cells that are similarly implicated in immune-mediated diseases. However, ILCs that promote the maintenance of intestinal epithelial cells, mostly through production of IL-22, also exist. ILCs rapidly respond to environmental cues, including cytokines, metabolic signals and luminal bacteria. They are potent and immediate producers of key cytokines linked to IBD pathogenesis, including TNF, IL-17, IL-22 and IFN-γ. Some subsets are implicated as mediators of chronic intestinal inflammation, whereas others might provide protective functions. They are present in the gut of patients with IBD and, intriguingly, closer scrutiny of IBD susceptibility loci shows that many of these genes are either expressed by, or are intimately linked to, ILC function. Looking forward, targeting ILCs could represent a new IBD treatment paradigm.


Modulation of excited-state intramolecular proton transfer by viscosity in protic media.
3-Hydroxyquinolones undergo excited-state intramolecular proton transfer (ESIPT), resulting in a dual emission highly sensitive to H-bonding perturbations. Here, we report on the strong effect of viscosity on the dual emission of 2-(2-thienyl)-3-hydroxyquinolone in protic solvents. An increase in viscosity significantly decreases the formation of the ESIPT product, thus changing dramatically the ratio of the two emission bands. Time-resolved studies suggest the presence of solvated species characterized by decay times close to the solvent relaxation times in viscous media. The intramolecular H bond in this species is probably disrupted by the solvent, and therefore, its ESIPT requires a reorganization of the solvation shell for restoring this intramolecular H bond. Thus, the ESIPT reaction of this dye and its dual emission depend on solvent relaxation rates and, therefore, on viscosity. The present results suggest a new physical principle for the fluorescence ratiometric measurement of local viscosity.


Hyperbaric oxygenation and antioxidant vitamin combination reduces ischemia-reperfusion injury in a rat epigastric island skin-flap model.
Reperfusion injury, caused by free oxygen radicals, is a chain of events that occurs in tissues exposed to a constant period of ischemia. The antioxidant vitamins E and C (VEC) and hyperbaric oxygenation (HBO) have beneficial effects in treating ischemic tissues following skin flap operations. In our study, we aimed to compare the effects of VEC and/or HBO in ischemia-reperfusion injury induced by free oxygen radicals in an experimental rat epigastric island skin-flap model. Eight hours of ischemia was provided by clamping the inferior epigastric pedicle following the flap elevation. The flap survey was determined to be 28.6% in controls, 59.2% in HBO group, 66.3% in VEC + HBO group, and 82% in VEC group (p < 0.05). We conclude that although HBO and/or VEC increased the flap viability significantly by reducing ischemia-reperfusion injury, the most promising results were obtained in the antioxidant vitamins group.


RNA-Pareto: interactive analysis of Pareto-optimal RNA sequence-structure alignments.
Incorporating secondary structure information into the alignment process improves the quality of RNA sequence alignments. Instead of using fixed weighting parameters, sequence and structure components can be treated as different objectives and optimized simultaneously. The result is not a single, but a Pareto-set of equally optimal solutions, which all represent different possible weighting parameters. We now provide the interactive graphical software tool RNA-Pareto, which allows a direct inspection of all feasible results to the pairwise RNA sequence-structure alignment problem and greatly facilitates the exploration of the optimal solution set.


[Comparative morphological investigations of human ovarian follicles and their oocytes (author's transl)].
During gynaecological operations on 27 women aged 20 to 52 years, all visible follicles were punctured and 42 oocytes removed for investigation partly by phase-contrast microscope and partly by semi-thin slides. specimens of the walls of the follicles WERE REMOVED AND EXAMINED HIstologically. Furthermore, the activity of steroid-3 beta-ol dehydrogenase was demonstrated histochemically in 14 cases. Comparative investigations of the walls of the follicles and their oocytes gave the following results: 1. In non-ovulatory tertiary follicles (3 to 12 mm in diameter) with no, or very little luteinization of the theca interna and a granulosa of 3 to 10 cell layers, steroid-3 beta-ol dehydrogenase activity was only found in the theca interna. In 5 out of 23 follicles, the oocytes showed early or advanced signs of degeneration. Thus, atresia of the follicles probably occurs after degeneration of the oocytes. 2. 12 follicles (8 to 30 mm in diameter) showed distinct luteinization of the theca and steroid-3 beta-ol dehydrogenase activity in the theca interna, as well as in the granulosa. 4 of these follicles (all at least 20 mm in diameter) showed oocytes with typical pre-ovulatory changes. In 3 other cases with a similar morphological picture of the follicular wall specimens, the oocytes showed no preovulatory changes. The conclusion is drawn that the functional differentiation of the wall of the follicle induces the preparation of the oocyte for ovulation. On the other hand, 5 similar follicles already had degenerating oocytes. It remains doubtful whether such follicles can reach ovulation. 3. In follicles with early or advanced atresia, the oocytes were completely degenerated.


Benchmark of correlation matrix renormalization method in molecule calculations.
We report benchmark calculations of the correlation matrix renormalization (CMR) approach for 23 molecules in the well-established G2 molecule set. This subset represents molecules with spin-singlet ground state in a variety of chemical bonding and coordination environments. The QUAsi-atomic minimal basis-set orbitals (QUAMBOs) are used as local orbitals in both CMR and full configuration interaction (FCI) calculations for comparison. The results obtained from the calculations are also compared with available experimental data. It is shown that the CMR method produces binding and dissociation energy curves in good agreement with the QUAMBO-FCI calculations as well as experimental results. The CMR benchmark calculations yield a standard deviation of 0.09 Å for the equilibrium bond length and 0.018 Hartree/atom for the formation energy, with a gain of great computational efficiency which scales like Hartree-Fock method.


Development of a care guidance index based on what matters to patients.
Targeting resources for a designated higher-risk subgroup is a strategy for chronic care management. However, risk-designation has several limitations: it is inaccurate, seldom helpful for care guidance, and potentially misallocates care away from many patients. To address limitations of risk designation, we tested a "what matters index" (WMI) in 19,593 adult patients with chronic conditions. The WMI contains five binary measures: insufficient confidence to manage health problems, level of pain, emotional problems, polypharmacy, and adverse medication effects. We examined its sum for association with patient-reported quality of life and prior emergency or hospital use. We compared its accuracy to a prototypic risk-designation model. The WMI was a good indicator for quality of life and in three diverse test populations it was strongly associated with the use of hospital and emergency services. For example, a sum of WMI ≥2 was associated with twice as many average uses as none; for ≥3, uses were three times higher. However, since relatively few patients use costly care, both the WMI and a prototypic risk-designation model had comparably low-positive predictive values. The WMI uses the patient voice to identify needs strongly associated with quality of life. Akin to risk designation models, the WMI can be used to place patients into groups associated with levels of costly services, but neither is likely to forecast costly service use for individuals. However, unlike risk-designation models, the WMI is based on measures that will immediately guide care for every patient.


Prospects of nano-material in breast cancer management.
Breast cancer evaluation and early diagnosis are core complexity worldwide and an ambiguity for scientists till date. Nano-materials are innovative tools for rapid diagnosis and therapy, which may induce an immense result in the field of oncology. Their exceptional size-dependent properties make them special and superior materials and quite indispensable in several fields of the human activities. The major obstacle in finding cure for malignant breast cancer is to increase in development of resistances for tumors to the therapeutic treatments. The widespread mammo-graph particle is being developed by nations to diagnosis disease in primitive stage to decline the mortality rates caused by breast carcinoma. The advancement of nano-particle based diagnostic tools facilitates in evaluation and provides encouraging development in breast cancer therapeutics. In this compact review, efforts have been made to compose the current advancements in the area of functional nano-particles. Furthermore, in vivo and in vitro applications of nano-materials in breast cancer management are also discussed.


Involvement of oxidative and L-arginine-NO pathways in the neurotoxicity of drugs of abuse in vitro.
1. Inhibitors of nitric oxide (NO) formation or ADP-ribosylation attenuate methamphetamine (METH)- and methylenedioxymetamphetamine (MDMA)-induced neurotoxicity on dopaminergic and serotonergic cells in primary cultures. 2. They also prevent METH-induced reactive gliosis in dopaminergic cultures. 3. Overexpression of superoxide dismutase (SOD) in cells obtained from SOD-transgenic mice also attenuates drug-induced toxicity. 4. These data indicate a role for oxygen-based and NO free radicals in the mechanisms of cell death associated with drugs of abuse in vitro.


Formation by the uterus of a peripheral layer of the sheath in microfilariae of Litomosoides carinii and Brugia malayi.
The eggshells of young developmental stages in the uterus are rather thin and homogenous. In the brezel stage of Brugia malayi they are 35 nm thick and 20 nm in Litomosoides carinii. In young developmental stages up to brezel stages the eggshells bind the lectins WGA, DBA and PNA labelled with colloidal gold. This shows that GlcNAc, GalNAc and Gal residues are present at the surface of the sheath. In intrauterine microfilariae of B. malayi the original sheath is reduced to a thickness of 7 nm. It is reinforced by secretions from a specialized area of the epithelium of the uterus which do not appear as a homogeneous layer but look like a string of pearls. This layer may be called the "uterine layer". It has a thickness of 40-80 nm. In the microfilaria of L. carinii, the thickness of the original sheath is reduced to 2-3 nm and the uterine layer has a thickness of 7 nm. The uterine layer does not react with any of the lectins, which shows that the surface lacks N-acetylglucosamine, N-acetylgalactosamine and galactose residues. The uterine layer appears to be an ancestral (plesiomorphic) feature which is present in free-living nematodes and the highly specialized bloodforms of filariae. The uterine layer seems to protect and disguise the original sheath against the immune reactions of the host.


Adenylate cyclase activation determines the effect of thromboxane synthase inhibitors on platelet aggregation in vitro. Comparison of platelets from responders and nonresponders.
The effect of five thromboxane-synthase inhibitors (UK-37248, UK-38485, UK-34787, CGS-13080 and OKY-1581) on arachidonic acid-induced platelet aggregation has been studied in vitro on platelets from 30 different healthy volunteers. The sensitivity of their platelets to adenylate cyclase stimulators or to dibutyryl cyclic AMP has been evaluated contemporarily. In 4 of the 30 volunteers tested no inhibition of platelet aggregation was obtained with any of the five thromboxane synthase inhibitors: these subjects were defined nonresponders; in 13 volunteers inhibition was observed with all the five drugs (responders). Significantly higher amounts of prostaglandin (PG)D2, prostacyclin and adenosine were required to suppress arachidonic acid-induced aggregation of platelets from nonresponders in vitro. No differences were instead observed between responders and nonresponders concerning platelet sensitivity to forskolin or dibutyryl cyclic AMP. The cyclic AMP rise obtained with exogenous prostacyclin was lower in platelets from nonresponders than in those from responders. PGE2 added in vitro to platelets from nonresponders exerted always a proaggregatory effect whereas this PG was antiaggregatory in most of the nonresponders. PGE2 blunted the antiaggregatory activity of PGD2 and limited the cyclic AMP increase induced by PGD2 in all the subjects tested. These data indicate that the unequal functional response of platelets from different subjects to thromboxane synthase inhibition depends essentially on adenylate cyclase function: a relative insensitivity of this enzyme to activating stimuli and the accumulation of substances (PGE2, PG endoperoxides, etc.) reducing the activity of adenylate cyclase may lead to continued platelet activation in some subjects despite the suppression of the synthesis of thromboxane A2.


Recurrence of Pilonidal Disease: Our Best is Not Good Enough.
Recurrence rates of pilonidal disease have been reported to be high as 30%. Patients with recurrent pilonidal disease often develop chronic wounds and draining sinuses that incur long-term morbidity, disability, and decreased quality of life. The aim of this study was to characterize rates of recurrence in patients with pilonidal disease treated by pediatric surgeons. A single center retrospective review of patients with pilonidal disease evaluated by pediatric surgeons from 2010 to 2015 was performed. Recurrence of pilonidal disease was defined as an episode of active pilonidal disease that required medical or surgical intervention >30 days from the preceding treatment. Repeated events proportional hazards regression modeling was performed to identify factors associated with time to recurrence. Among 307 patients treated for pilonidal disease, nearly 50% were male, and the median age at initial evaluation was 16 years (IQR 15-17). Approximately 45% were obese (BMI ≥ 95th percentile). The initial treatment during the study period was surgical excision in two-thirds and incision and drainage and/or antibiotics in one-third. The overall recurrence rate was 33%, with the majority of recurrences (80%) occurring within the first year. On multivariable analysis, obese BMI was the only factor independently associated with time to disease recurrence. Pilonidal disease has a substantial recurrence rate even after surgical excision. Future studies investigating treatments that can prevent disease recurrence are needed.


Group I mGluR5 metabotropic glutamate receptors regulate proliferation of neuronal progenitors in specific forebrain developmental domains.
Major classical neurotransmitters including GABA and glutamate play novel morphogenic roles during development of the mammalian CNS. During forebrain neurogenesis, glutamate regulates neuroblast proliferation in different germinal domains using receptor subtype-specific mechanisms. For example, ionotropic N-methyl-D-aspartate (NMDA) or alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) glutamate receptors mediate distinct proliferative effects in ventral or dorsal forebrain germinal domains, and regulate the correct number of neurons that populate the striatum or cerebral cortex. Recent work suggests metabotropic receptors may also mediate glutamate's proliferative effects. Group I mGluR5 receptor subtypes are highly expressed in forebrain germinal zones. Using in vitro and in vivo methods, we demonstrate mGluR5 receptor activation plays an important role in neuroblast proliferation in the ventral telencephalon, and helps determine the complement of striatum projection neurons. mGluR5 receptor-mediated effects on striatal neuronal progenitors are restricted mainly to early cycling populations in the ventricular zone, with little effect on secondary proliferative populations in the subventricular zone. In contrast to proliferative effects in the ventral telencephalon, mGluR5 receptors do not modulate proliferation of dorsal telencephalon-derived cortical neuroblasts. Heterogeneous domain-specific proliferative effects of glutamate-mediated by specific receptor subtypes provide an important developmental mechanism allowing generation of the correct complement of neuronal subtypes that populate the mammalian forebrain.


Relapsing polychondritis.
Relapsing polychondritis is a progressive multisystem disorder that predominantly affects cartilage, producing an inflammatory reaction. We present case histories from four patients having mainly respiratory symptoms in whom we measured the coronal diameters of the trachea and compared them with the tracheal measurements from radiographs of patients with no respiratory symptoms. The results showed that the disease produced diffuse narrowing of the main airway in all four patients. Early detection of tracheal narrowing is important, as patients presenting with respiratory tract disease have a worse prognosis than those who develop it later in the course of their illness. Other features of the disease are also discussed with reference to the cases presented.


Cytoskeleton structure, pattern of mitochondrial activity and ultrastructure of frozen or vitrified sheep embryos.
Even though sheep embryo cryopreservation is a commonly used procedure the survival and pregnancy outcomes can vary greatly. This study investigated whether cryopreservation was causing subtle changes in ultrastructure, mitochondrial activity or cytoskeletal integrity. Sheep embryos were either slow cooled in 1.5 M EG (n = 22), or vitrified in 20% EG + 20% DMSO with 0.5 M sucrose in Open Pulled Straws (OPS) (n = 24). One hour after warming the cryopreserved embryos differed from control embryos in that they had no mitochondrial activity combined with cytoskeletal disorganization and large vesicles. Vitrified embryos also showed many points of cytoskeleton disruption. Ultrastructural alterations resulting from actin filaments disorganization were observed in both cryopreserved groups. This includes areas presenting no cytoplasmic organelles, Golgi complex located far from the nucleus and a decrease of specialized intercellular junctions. Additionally, large vesicles were observed in vitrified morulae and early blastocysts. The alterations after cryopreservation were proportional to embryo quality as assessed using the stereomicroscope. Even in the absence of mitochondrial activity, grade I and II cryopreserved embryos contained mitochondria with normal ultrastructure. Embryos classified as grade I or II in the stereomicroscope revealed mild ultrastructural alterations, meaning that this tool is efficient to evaluate embryos after cryopreservation.


Relationship between short wavelength perimetry and central corneal thickness values in ocular hypertensive subjects.
To evaluate the results of short wavelength perimetry (SWAP) of ocular hypertensive (OHT) patients and correlate these findings with central corneal thickness (CCT) measurements. Thirty-seven OHT patients with a mean age of 50.2+/-8.2 (SD) years and 30 control subjects with a mean age of 50.3+/-8.5 (SD) years were included in this study. A questionnaire was applied to patients to evaluate the demographic risk factors that may predict glaucoma development. After a detailed ophthalmologic examination, achromatic and short wavelength perimetries and ultrasonic pachymetry were performed and the results were compared between the two groups with Student t test and Mann-Whitney U test. A p value<0.05 is considered as statistically significant. Mean CCT was higher in the OHT group (right eye; 558.13+/-28.39 microm and left eye; 558.94+/-27.30 microm) when compared with the control subjects (524.66+/-30.53 microm and 525.86+/-30.46 microm, respectively) (p<0.01). A significant positive correlation was found between CCT measurements and intraocular pressure (r=0.5, p<0.001). Four right eyes (10.8%) and five left eyes (13.5%) of OHT patients had defects in SWAP. OHT patients with SWAP abnormalities had significantly lower CCT measurements in right (527.25+/-17.34 microm) and left eye (528.80+/-13.60 microm) when compared with OHT patients without SWAP defects (561.87+/-27.29 microm and 563.65+/-25.92 microm, respectively) (p<0.05). Significant correlations were found between CCT and SWAP MD, PSD, and CPSD (p<0.05). OHT patients with SWAP abnormalities had significantly lower CCT measurements than those without. CCT is considered as a risk factor for the development of glaucomatous damage in OHT patients.


Uncleaved legumin in developing maize endosperm: identification, accumulation and putative subcellular localization.
While identifying proteins present in the cytoskeleton and protein body fractions from maize (Zea mays L.) endosperm, a 51 kDa protein was discovered in a fraction containing small (approximately 200 nm in diameter) protein bodies. Based on partial amino acid sequences of V8 protease fragments, degenerate primers were made and fragments of cDNA encoding these partial sequences were cloned. Using 3' and 5' PCR, a full-length cDNA encoding this 51 kDa protein was obtained, which was identified as legumin-1. In other plants, this protein is generally cleaved into 20 and 35 kDa subunits after synthesis. However, SDS-PAGE of both the native and denatured protein indicates that cleavage does not occur in corn endosperm, even though the cleavage site (asparagine) is conserved. The lack of cleavage is presumably because the canonical cleavage sequence downstream from the cleavage site is almost totally absent. levels of transcript and encoded protein were compared in all three varieties and it was shown that both are more abundant in wild-type maize than in opaque-2 or sweet corn. Finally, using TEM, it was shown that the protein apparently occurs in morphologically distinct protein bodies, very similar to the protein bodies in legumes.


Identification of novel sources of host plant resistance to known soybean aphid biotypes.
While soybean cultivars with resistance to the soybean aphid (Aphis glycines Matsumura) have been commercially released, the presence of virulent biotypes capable of overcoming plant resistance threatens the durability of host plant resistance as a stable management tactic. Novel sources of host plant resistance are needed to combat rapid biotype evolution. In this study, we screened 1,061 soybean plant introductions (PIs) for resistance to three known biotypes of A. glycines. Based on a series of growth chamber and field screenings, we identified 11 PIs that showed resistance to biotype 1 of A. glycines. Among these 11 PIs, 7 PIs were resistant to biotype 2 and 5 PIs were resistant to biotype 3. Further, two PIs (PI 606390A from Vietnam and PI 340034 from South Korea) showed resistance to all three biotypes of A. glycines. We also identified 11 PIs that were potentially tolerant to A. glycines, illustrated by no adverse impact on plant quality because of A. glycines infestation. As resistant PIs identified in this study belong to maturity group II-IV, they can be readily crossed to early maturing cultivars adapted to north-central states of the United States, where A. glycines is a major pest. The genetic characterization of resistance in these PIs and incorporation of novel resistant genes into elite soybean cultivars will broaden the defense against multiple biotypes of A. glycines.


Role of alpha adrenergic blocker in the management of posterior urethral valves.
To evaluate the effect of Terazosin (alpha1 adrenergic blocker) on bladder emptying in children with posterior urethral valves. Forty-two children with significant post void residual urine after valves ablation were placed on Terazosin ranging from 0.25 to 2 mg. Post void urine at the commencement and at follow up was monitored with abdominal ultrasound. Post void residual urine significantly reduced in 40 patients (95%) who were put on Terazosin. Mean pretreatment PVR was 15.7 ml and mean PVR at the last follow up was 2.4 ml (P = 0.000). This was a reduction of 85% in the pretreatment post void residual urine volume. All the patients had improvement in urinary stream. One patient reacted to Terazosin with hypotension necessitating its withdrawal. Mean follow up was 17 months. Terazosin has proved to be safe and results in significant improvement in bladder emptying in our patients with posterior urethral valves. Randomized controlled trial and long-term follow up are necessary to further define the role of alpha1 adrenergic blocker therapy in children with posterior urethral valves. This study will become the justification for such a study.


Spontaneous outgrowth of Epstein-Barr virus-positive B-cell lines from circulating human B cells of different buoyant densities.
Epstein-Barr virus (EBV) has potent cell-growth-transforming activity for human B lymphocytes in vitro, yet appears to persist in the circulating B-cell pool of virus carriers in vivo as a largely asymptomatic (i.e., non-growth-transforming) infection. The true nature of this infection, and the identity of the cells involved, remain to be determined. Studies of Lewin et al. (1987) have suggested (i) that the frequency of virus-infected cells in the circulating B-cell pool differs in different buoyant density fractions, being most abundant in the low-density population, and (ii) that rare virus-infected cells with the capacity for direct in vitro outgrowth to EBV-transformed cell lines are segregated within the high-density population. We have repeated this work using B-cell fractions from a much larger panel of asymptomatic virus carriers and find (i) that the incidence of virus-infected B cells is not significantly different between high- and low-density fractions, and (ii) that virus-infected cells from both fractions give rise to EBV-transformed cell lines in culture predominantly through a 2-step mechanism of virus replication and secondary infection rather than by direct outgrowth.


Bacterial Colonization in the Marginal Region of Ceramic Restorations: Effects of Different Cement Removal Methods and Polishing.
This study evaluated the effects of excess cement removal techniques, with or without subsequent polishing, on biofilm formation and micromorphology in the marginal region of the tooth/restoration. From bovine teeth, 96 dentin blocks (4 × 8 × 2 mm) were produced, molded, and reproduced in type IV gypsum, on which 96 pressed ceramic blocks (Vita PM9, Vita Zahnfabrik; 4 × 8 × 2 mm) were produced via the lost wax technique. The dentin blocks and their respective ceramic blocks were cemented with a self-adhesive resin cement (RelyX U200, 3M ESPE), and cement excess was removed from the margin using four different techniques, followed or not by polishing with silicone rubber tips: MBr, removal with microbrush and photoactivation; MBr-Pol, MBr + polishing; Br, removal with brush and photoactivation; Br-Pol, Br + polishing; Photo-Expl, 5 seconds of initial photoactivation, removal with explorer, and final curing; Photo-Expl-Pol, Photo-Expl + polishing; Photo-SB, 5 seconds of initial photoactivation, removal with scalpel, and final curing; and Photo-SB-Pol, Photo-SB + polishing. After 24 hours, the roughness in the marginal region was analyzed using a profilometer (three measurements on each sample). Micromorphological analyses of the region were performed by stereomicroscope and scanning electron microscopy (SEM). Then the samples were contaminated with sucrose broth standardized suspension with Streptococcus mutans , Staphylococcus aureus , and Candida albicans and incubated for a period of 48 hours. The samples were quantitatively analyzed for bacterial adherence in the marginal region by confocal laser scanning microscopy and counting of colony-forming units (CFUs/mL) and qualitatively analyzed using SEM. Roughness data (Ra) were submitted to two-way analysis of variance, Tukey test at a confidence level of 95%, and Student t-tests. CFU, biomass, and biothickness data were analyzed by Kruskal-Wallis, Mann-Whitney, and Dunn tests. The removing technique statistically influenced Ra (MBr, p=0.0019; Br, p=0.002; Photo-Expl, p=0.0262; Photo-SB, p=0.0196) when comparing the polished and unpolished groups. The MBr and MBr-Pol technique differed significantly for CFU/mL values (p=0.010). There was no significant difference in the amounts of biomass and biothickness comparing polished and unpolished groups and when all groups were compared (p>0.05). Different morphological patterns were observed (more regular surface for polished groups). We conclude that margin polishing after cementation of feldspar/pressed ceramic restorations is decisive for achieving smoother surfaces, as the excess cement around the edges can increase the surface roughness in these areas, influencing bacterial adhesion.


An innovative approach to reducing medical care utilization and expenditures.
In a retrospective study, we assessed the impact on medical utilization and expenditures of a multicomponent prevention program, the Maharishi Vedic Approach to Health (MVAH). We compared archival data from Blue Cross/Blue Shield Iowa for MVAH (n = 693) with statewide norms for 1985 through 1995 (n = 600,000) and with a demographically matched control group (n = 4,148) for 1990, 1991, 1994, and 1995. We found that the 4-year total medical expenditures per person in the MVAH group were 59% and 57% lower than those in the norm and control groups, respectively; the 11-year mean was 63% lower than the norm. The MVAH group had lower utilization and expenditures across all age groups and for all disease categories. Hospital admission rates in the control group were 11.4 times higher than those in the MVAH group for cardiovascular disease, 3.3 times higher for cancer, and 6.7 times higher for mental health and substance abuse. The greatest savings were seen among MVAH patients older than age 45, who had 88% fewer total patients days compared with control patients. Our results confirm previous research supporting the effectiveness of MVAH for preventing disease. Our evaluation suggests that MVAH can be safely used as a cost-effective treatment regimen in the managed care setting.


A single [3H]thymidine-based limiting dilution analysis to determine HTLp and CTLp frequencies for bone marrow donor selection.
Histocompatibility between recipient and donor is a critical factor in allogeneic BMT which, to a large extent, determines the incidence of GVHD after BMT. Functional histocompatibility assays, such as the helper T lymphocyte precursor frequency assay (HTLp) and the cytotoxic T lymphocyte precursor frequency assay (CTLp), have proved to be helpful tools in facilitating donor selection procedures. However, a major drawback of these assays is that they are laborious and require large numbers of cells. We therefore adapted a [3H]thymidine-based assay, the 'JAM' test, as a read-out for CTLp frequencies, to replace the more cumbersome 51Cr-release assay. Furthermore, we applied an experimental setup that enables the assessment of HTLp and CTLp frequencies from a single limiting dilution assay to reduce the number of cells needed. The newly developed assay is relatively easy to perform and has the advantage that different subsets of T cells can be quantified in a single ongoing alloreaction. When the combined assay was applied in unrelated donor selection it proved to be a sensitive method that enables differentiation in suitability of distinct donors for a single patient. Therefore, the combined HTLp/CTLp assay appears to be a practical and sensitive method for identifying functional histocompatibility in related and unrelated donor/recipient combinations.


Multicompartmental lipid-protein nanohybrids for combined tretinoin/herbal lung cancer therapy.
Aim: Multicompartmental lipid-protein nanohybrids (MLPNs) were developed for combined delivery of the anticancer drugs tretinoin (TRE) and genistein (GEN) as synergistic therapy of lung cancer. Materials & methods: The GEN-loaded lipid core was first prepared and then coated with TRE-loaded zein shell via nanoprecipitation. Results: TRE/GEN-MLPNs demonstrated a size of 154.5 nm. In situ ion pair formation between anionic TRE and the cationic stearyl amine improved the drug encapsulation with enhanced stability of MLPNs. TRE/GEN-coloaded MLPNs were more cytotoxic against A549 cancer cells compared with combined free GEN/TRE. In vivo, lung cancer bearing mice treated with TRE/GEN-MLPNs displayed higher apoptotic caspase activation compared with mice-treated free combined GEN/TRE. Conclusion: TRE/GEN-MLPNs might serve as a promising parenteral nanovehicles for lung cancer therapy.


Enhanced frequency of spontaneous B cell lines from Epstein-Barr virus (EBV) seropositive donors 80 years and older.
Based on the well-recognized decline in immunocompetence which develops with advancing age, we have evaluated the effect of age on the frequency of development of spontaneous Epstein-Barr virus (EBV)-infected B cell lines. Blood mononuclear cells were isolated from 38 clinically healthy seropositive donors. The cells were maintained in vitro according to routine culture conditions for lymphocytes. Eight spontaneously EBV-infected B lymphoblastoid cell lines (LCL) were isolated. The LCL developed in 12.5, 14.3, or 6.3% of the samples derived from donors in the three age groups 20-39, 40-59, 60-79, respectively. In contrast, samples from five of seven (71%) donors 80 years and older yielded LCL. Although the reason(s) for the increased frequency of occurrence of spontaneous LCL from the older adults is yet to be explored, the possible role of the virus-specific T lymphocytes as a contributing factor is discussed.


Long-term safety and feasibility of three-vessel multimodality intravascular imaging in patients with ST-elevation myocardial infarction: the IBIS-4 (integrated biomarker and imaging study) substudy.
We assessed the feasibility and the procedural and long-term safety of intracoronary (i.c) imaging for documentary purposes with optical coherence tomography (OCT) and intravascular ultrasound (IVUS) in patients with acute ST-elevation myocardial infarction (STEMI) undergoing primary PCI in the setting of IBIS-4 study. IBIS4 (NCT00962416) is a prospective cohort study conducted at five European centers including 103 STEMI patients who underwent serial three-vessel coronary imaging during primary PCI and at 13 months. The feasibility parameter was successful imaging, defined as the number of pullbacks suitable for analysis. Safety parameters included the frequency of peri-procedural complications, and major adverse cardiac events (MACE), a composite of cardiac death, myocardial infarction (MI) and any clinically-indicated revascularization at 2 years. Clinical outcomes were compared with the results from a cohort of 485 STEMI patients undergoing primary PCI without additional imaging. Imaging of the infarct-related artery at baseline (and follow-up) was successful in 92.2% (96.6%) of patients using OCT and in 93.2% (95.5%) using IVUS. Imaging of the non-infarct-related vessels was successful in 88.7% (95.6%) using OCT and in 90.5% (93.3%) using IVUS. Periprocedural complications occurred <2.0% of OCT and none during IVUS. There were no differences throughout 2 years between the imaging and control group in terms of MACE (16.7 vs. 13.3%, adjusted HR1.40, 95% CI 0.77-2.52, p = 0.27). Multi-modality three-vessel i.c. imaging in STEMI patients undergoing primary PCI is consistent a high degree of success and can be performed safely without impact on cardiovascular events at long-term follow-up.


Dual-energy compared to single-energy CT in pediatric imaging: a phantom study for DECT clinical guidance.
Dual-energy CT technology is available on scanners from several vendors and offers significant advantages over classic single-energy CT technology in multiple clinical applications. Many studies have detailed dual-energy CT applications in adults and several have evaluated the relative radiation dose performance of dual-energy CT in adult imaging. However, little has been published on dual-energy CT imaging in the pediatric population, and the relative dose performance of dual-energy CT imaging in the pediatric population is not well described. When evaluating dual-energy CT technology for implementation into a routine clinical pediatric imaging practice, the radiation dose implications must be considered, and when comparing relative CT dose performance, image quality must also be evaluated. Therefore the purpose of this study is to develop dual-energy CT scan protocols based on our optimized single-energy scan protocols and compare the dose. We scanned the head, chest and abdomen regions of pediatric-size anthropomorphic phantoms with contrast inserts, using our optimized single-energy clinical imaging protocols on a Siemens Flash® CT scanner. We then scanned the phantoms in dual-energy mode using matching image-quality reference settings. The effective CT dose index volume (CTDIvol) of the scans was used as a surrogate for relative dose in comparing the single- and dual-energy scans. Additionally, we evaluated image quality using visual assessment and contrast-to-noise ratio. Dual-energy CT scans of the head and abdomen were dose-neutral for all three phantoms. Dual-energy CT scans of the chest showed a relative dose increase over the single-energy scan for 1- and 5-year-old child-based age-equivalent phantoms, ranging 11-20%. Quantitative analysis of image quality showed no statistically significant difference in image quality between the single-energy and dual-energy scans. There was no clinically significant difference in image quality by visual assessment. Dual-energy CT is dose-neutral in imaging the head and abdomen in children. It is not dose-neutral in chest imaging of very small children. With a better understanding of the dose consequences of converting single-energy protocols to dual-energy protocols we can begin to implement clinical dual-energy CT and utilize its unique capabilities in pediatric imaging.


A response to "Likelihood ratio as weight of evidence: A closer look" by Lund and Iyer.
Recently, Lund and Iyer (L&I) raised an argument regarding the use of likelihood ratios in court. In our view, their argument is based on a lack of understanding of the paradigm. L&I argue that the decision maker should not accept the expert's likelihood ratio without further consideration. This is agreed by all parties. In normal practice, there is often considerable and proper exploration in court of the basis for any probabilistic statement. We conclude that L&I argue against a practice that does not exist and which no one advocates. Further we conclude that the most informative summary of evidential weight is the likelihood ratio. We state that this is the summary that should be presented to a court in every scientific assessment of evidential weight with supporting information about how it was constructed and on what it was based.


Semiconductor quantum templates: bottom-up design of optical devices and photonic circuits using sub-nanoscale high monolayer features and quantum optics.
We propose an alternative bottom-up technique for designing, fabricating and monolithically integrating optical components, including functional distributed feedback lasers, modulators, waveguides, etc in a single semiconductor wafer without any need for etching or post-processing epitaxial growth. The proposed technique is based on the formation of semiconductor quantum templates at the well/barrier interfaces of quantum well structures. Such templates are responsible for changing the thickness of a quantum well in designated regions by adding one extra monolayer of the well material in those regions during the growth process. We show that, using a control laser field, these templates or sub-nanoscale high monolayer features allow us to spatially control the formation of electromagnetically induced transparency, gain without inversion, and coherent enhancement/suppression of the refractive index along the plane of the quantum well. We demonstrate that this can lead to bottom-up design capabilities for functional optical devices and their monolithic integration using a single epitaxial growth process.


One-class acoustic characterization applied to blood detection in IVUS.
Intravascular ultrasound (IVUS) is an invasive imaging modality capable of providing cross-sectional images of the interior of a blood vessel in real time and at normal video framerates (10-30 frames/s). Low contrast between the features of interest in the IVUS imagery remains a confounding factor in IVUS analysis; it would be beneficial therefore to have a method capable of detecting certain physical features imaged under IVUS in an automated manner. We present such a method and apply it to the detection of blood. While blood detection algorithms are not new in this field, we deviate from traditional approaches to IVUS signal characterization in our use of 1-class learning. This eliminates certain problems surrounding the need to provide "foreground" and "background" (or, more generally, n-class) samples to a learner. Applied to the blood-detection problem on 40 MHz recordings made in vivo in swine, we are able to achieve approximately 95% sensitivity with approximately 90% specificity at a radial resolution of approximately 600 microm.


Evaluation of copper-1,3,5-benzenetricarboxylate metal-organic framework (Cu-MOF) as a selective sorbent for Lewis-base analytes.
The metal-organic framework copper-1,3,5-benzenetricarboxylate (Cu-BTC) was evaluated for its ability to selectively interact with Lewis-base analytes by examining retention on gas-chromatographic columns packed with Chromosorb W HP that contained 3.0% SE-30 along with various loadings of Cu-BTC. Scanning electron microscopy images of the support material showed the characteristic Cu-BTC crystals embedded in the SE-30 coating on the diatomaceous support. The results indicated that the Cu-BTC-containing stationary phase had limited thermal stability (220 °C) and strong general retention for analytes. Kováts index calculations showed selective retention (amounting to about 300 Kováts units) relative to n-alkanes for many small Lewis-base analytes on a column that contained 0.75% Cu-BTC compared with an SE-30 control. Short columns that contained lower loadings of Cu-BTC (0.10%) allowed elution of nitroaromatics; however, selectivity was not observed for aromatic compounds (including nitroaromatics) or nitroalkanes. Observed retention characteristics are discussed.


Overproduction of soluble trichodiene synthase from Fusarium sporotrichioides in Escherichia coli.
Trichodiene synthase is a sesquiterpene cyclase isolated from various fungal species which catalyzes the cyclization of farnesyl diphosphate (FPP) to trichodiene. The trichodiene synthase gene (Tox5) of Fusarium sporotrichioides has previously been cloned and expressed as 0.05-0.1% of total cell protein in Escherichia coli. We have used polymerase chain reaction to amplify the trichodiene coding sequence carried on the plasmid pTS56-1. The resulting DNA, carrying a BamHI restriction site and the T7 gene 10 ribosome binding site and translational spacer element immediately upstream of the ATG start codon as well as a HindIII site adjacent to the translational stop codon, was inserted into the corresponding sites of the expression vector pLM1. The latter vector carried the promoter and translational leader sequence from T7 gene 10 and the E. coli rmBT1T2 tandem transcription terminator. This construct was cloned into E. coli BL21 (DE3). The resulting transformants, when induced with isopropyl beta-D-thiogalactoside, produced trichodiene synthase as 20-30% of total soluble protein. The recombinant synthase, which could be purified five-fold to homogeneity by ammonium sulfate precipitation, ion-exchange chromatography on Q Sepharose, and gel filtration on Superose 12, was identical to native protein in steady-state kinetic parameters and mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had the expected MENFP N-terminal sequence.


Preoperative diagnosis of 221 consecutive ovarian masses: scoring system and expert evaluation.
The aim of our work was to assess the diagnostic accuracy of a scoring system versus subjective assessment of the risk of malignancy of pelvic masses achieved by gynecologist/sonologists in the preoperative triage of a busy gynecology department. One hundred and eighty-two consecutive patients who underwent surgical removal of ovarian neoplasms were examined. In 39 patients pelvic masses were bilateral. The total number of neoplasms analyzed in this series was 221. Lesions were examined and scored according to the sonographic characteristics. Gynecologist/sonologists also recorded a subjective evaluation of the adnexal masses defining them as "probably benign", and "suspicious/probably malignant". Preoperative ultrasound risk assessment was compared to the final pathologic report and diagnostic accuracy was calculated. CA125 was obtained in all patients and its independent and combined accuracy was calculated. The sensitivity of the scoring system and subjective evaluation was 86% and 95% respectively, specificity was 79% and 91% with a positive predictive value of 41% and 53%. The frequency of positive diagnoses for CA125 was 44% due to the high prevalence of endometrosis in this series (48%). In premenopausal patients (75%) the specificity was 93% and 89% for scoring and subjective evaluation, respectively. The combined use of morphological scoring and CA125 achieved higher specificity and positive predictive values both for the whole series and in premenopausal patients. These results confirmed that the experience of gynecological surgeons with ultrasound skills, outperforms the morphological indexing assessment of ovarian masses. Nonetheless an easy sonographic descriptive scoring system is not significantly lower in accuracy than the expertise achieved by gynecologists with sonographic skills.


Mapping change in human pressure globally on land and within protected areas.
It is widely accepted that the main driver of the observed decline in biological diversity is increasing human pressure on Earth's ecosystems. However, the spatial patterns of change in human pressure and their relation to conservation efforts are less well known. We developed a spatially and temporally explicit map of global change in human pressure over 2 decades between 1990 and 2010 at a resolution of 10 km(2) . We evaluated 22 spatial data sets representing different components of human pressure and used them to compile a temporal human pressure index (THPI) based on 3 data sets: human population density, land transformation, and electrical power infrastructure. We investigated how the THPI within protected areas was correlated to International Union for Conservation of Nature (IUCN) management categories and the human development index (HDI) and how the THPI was correlated to cumulative pressure on the basis of the original human footprint index. Since the early 1990s, human pressure increased 64% of the terrestrial areas; the largest increases were in Southeast Asia. Protected areas also exhibited overall increases in human pressure, the degree of which varied with location and IUCN management category. Only wilderness areas and natural monuments (management categories Ib and III) exhibited decreases in pressure. Protected areas not assigned any category exhibited the greatest increases. High HDI values correlated with greater reductions in pressure across protected areas, while increasing age of the protected area correlated with increases in pressure. Our analysis is an initial step toward mapping changes in human pressure on the natural world over time. That only 3 data sets could be included in our spatio-temporal global pressure map highlights the challenge to measuring pressure changes over time.


Time-Dependent Diffusion in Prostate Cancer.
Prior studies in prostate diffusion-weighted magnetic resonance imaging (MRI) have largely explored the impact of b-value and diffusion directions on estimated diffusion coefficient D. Here we suggest varying diffusion time, t, to study time-dependent D(t) in prostate cancer, thereby adding an extra dimension in the development of prostate cancer biomarkers. Thirty-eight patients with peripheral zone prostate cancer underwent 3-T MRI using an external-array coil and a diffusion-weighted image sequence acquired for b = 0, as well as along 12 noncollinear gradient directions for b = 500 s/mm using stimulated echo acquisition mode (STEAM) diffusion tensor imaging (DTI). For this sequence, 6 diffusion times ranging from 20.8 to 350 milliseconds were acquired. Tumors were classified as low-grade (Gleason score [GS] 3 + 3; n = 11), intermediate-grade (GS 3 + 4; n = 16), and high-grade (GS ≥4 + 3; n = 11). Benign peripheral zone and transition zone were also studied. Apparent diffusion coefficient (ADC) D(t) decreased with increasing t in all zones of the prostate, though the rate of decay in D(t) was different between sampled zones. Analysis of variance and area under the curve analyses suggested better differentiation of tumor grades at shorter t. Fractional anisotropy (FA) increased with t for all regions of interest. On average, highest FA was observed within GS 3 + 3 tumors. There is a measurable time dependence of ADC in prostate cancer, which is dependent on the underlying tissue and Gleason score. Therefore, there may be an optimal selection of t for prediction of tumor grade using ADC. Controlling t should allow ADC to achieve greater reproducibility between different sites and vendors. Intentionally varying t enables targeted exploration of D(t), a previously overlooked biophysical phenomenon in the prostate. Its further microstructural understanding and modeling may lead to novel diffusion-derived biomarkers.


Higher incidence of Gastrospirillum sp. in swine with gastric ulcer of the pars oesophagea.
Gastric ulcer in swine is characterized by an area of acid-peptic digestion, occurs usually in the pars oesophagea of the stomach, and has unknown etiopathogenesis. The present work was carried out to investigate the prevalence of the newly described spiral-shaped microorganism Gastrospirillum sp. ("Gastrospirillum suis") in stomachs of abattoir pigs with and without gastric ulcer. Stomachs were removed from 32 consecutive pigs presenting apparently normal mucosa and from 32 additional consecutive pigs presenting frank, chronic gastric ulcer of the pars oesophagea. Fragments of antral, oxyntic, cardiac and pars oesophagea regions were taken from each stomach and processed for histology and for identification of Gastrospirillum sp. in tissue sections. The microorganisms were identified mainly in the mucous layer and in gastric foveolas of the antral and oxyntic mucosa. Forty pigs (62.5%) were positive for Gastrospirillum sp.; among them, 27 (67.5%) had gastric ulcer, and 13 (32.5%) had no ulcer. Twenty-four pigs (37.5%) were negative for Gastrospirillum sp.; among them, five (20.8%) presented with gastric ulcer, and 19 (79.2%) had no ulcer. There was a significant difference between pigs with and without gastric ulcer in regard to the presence of Gastrospirillum sp. (P < 0.01). The spiral-shaped microorganism Gastrospirillum sp. that inhabits the stomach of pigs should be considered a possible factor connected with the etiopathogenesis of swine gastric ulcer.


Astigmatic keratotomy: a review of basic concepts with case reports.
Despite the use of small incisions, better wound closure, and other technical improvements in cataract and corneal surgery, astigmatism remains a problem for the cataract and refractive surgeon. Case reports illustrate the applicability of astigmatic keratotomy in post-cataract and idiopathic astigmatism. The basic concepts of corneal relaxing incisions are reviewed and the frequently misunderstood coupling phenomenon is explained in the light of the "law of elastic domes." Properly used, astigmatic keratotomy with coupling effectively corrects a wide variety of types of astigmatism.


Extra-osseous fat fluid level: a specific sign for osteomyelitis.
Osteomyelitis may pose as a diagnostic dilemma on various imaging modalities and may be confused with neoplasms and other pathology. Although a rare finding, extra-osseous fat fluid level, especially when associated with spongy bone destruction, can be considered a specific sign of osteomyelitis. Previously, only two cases of extra-osseous fat fluid level in osteomyelitis have been reported, one on computed tomography (CT) and the other on magnetic resonance imaging (MRI). The former was a case of septic arthritis with intra-articular fat fluid level. A case of osteomyelitis is presented with the demonstration of extra-osseous fat fluid level. Our case is unique in providing exquisite CT and MRI correlation.


Do clinic-based STD data reflect community patterns?
Sexually transmitted disease (STD) interventions in communities are often based on studies conducted in STD clinics. Clinic-based studies are relatively easy to conduct, but they do not include all people at risk for STDs in the community. Although it is widely believed that clinic-based data present a biased picture of the community, the differences between clinical and community perspectives seldom have been quantified. The Sexually Transmitted Epidemic Prevention (STEP) Project included in-depth standardized interviews administered in an STD clinic and the neighbourhoods of the same town. The populations could be quantitatively compared on some questions. The two samples yielded similar estimates of the proportions using condoms, the average age at first intercourse, and the proportion sexually abused as a child. Risky sexual behaviours were more common in the clinic population, however, and the community sample was more socio-economically stable. Clinic-based data may be useful for targeting high risk people in the community but less useful for reaching those at lower risk. The utility of clinic-based studies for community-based interventions may be enhanced by collecting extra information from clinic patients with lower risk profiles.


Treatment adherence and long-term outcomes.
The successful management of schizophrenia is an enormous public health issue. Although antipsychotic medications can be very helpful in reducing rates of relapse and rehospitalization, nonadherence to medication is a frequent cause of exacerbations in psychopathology, psychotic relapse, and rehospitalization. Relapses can have devastating consequences in a variety of clinical and functional domains. Nonadherence can result from a variety of factors that vary from patient to patient and vary over time in individual patients. A number of strategies have been developed to assess and facilitate adherence. The first critical step is clinician awareness of the scope of the problem and consideration of appropriate strategies to address it.


Through which looking glass? Distinct sources of public regard and self-esteem among first- and second-generation immigrants of color.
How we think other people see us has long been believed to influence how we see ourselves, a phenomenon known as reflected appraisal. Among members of ethnic minority groups and immigrant groups, however, inconsistent patterns have been found. This study examined how immigrants' views of their ethnic group and of themselves are tied the appraisals of specific others. We hypothesized that members of the heritage culture and of White Americans are implicated in the reflected appraisal process to differing degrees. Moreover, we hypothesized that immigrants (first-generation) and the children of immigrants (second-generation) are affected differently by the appraisals of White Americans and heritage culture members. Results showed that perceived appraisal by members of the heritage culture was linked to how immigrants felt about their group and to their personal self-esteem. Perceived appraisal of White Americans was not related to how immigrants felt about their group and was only weakly related to personal self-esteem. The results highlight the distinction between public regard from the heritage culture and from White Americans and underscore the importance of the former for immigrants' private regard and self-esteem.


A gel transfer tank for immunoblotting and its application for analysis of nuclear protein antigens.
The design of a gel transfer tank for immunoblotting is described. It is simple and cheap to make, provides a uniform field and uniform transfer over the whole area of the gel, and can easily be adapted for use with any size of gel. It has been used for transfer of proteins from both sodium dodecyl sulfate-polyacrylamide and isoelectric focusing gels to nitrocellulose membranes and its application to the analysis of nuclear proteins is described.


Human herpesvirus-8-encoded signalling ligands and receptors.
Analysis of the genome of human herpesvirus 8 (HHV-8) led to the discovery of several novel genes, unique among the characterized gammaherpesviruses. These include cytokines (interleukin-6 and chemokine homologues), two putative signal-transducing transmembrane proteins encoded by genes K1 and K15 at the genome termini, and an OX-2 (CD200) receptor homologue that had not previously been identified in a gammaherpesvirus. HHV-8 also specifies a diverged version of the gammaherpesvirus-conserved G protein-coupled chemokine receptor (vGCR) and a latently expressed protein unique to HHV-8 specified by open reading frame (ORF) K12. These cytokine and receptor homologues mediate signal transduction or modulate the activities of other endogenous cytokines and receptors to enhance viral productive replication, regulate latent-lytic switching, evade host attack, or mediate cell survival. The viral signalling ligands and receptors are also potential contributors to virus-associated diseases, Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease, and so represent potentially important targets for therapeutic and antiviral drugs. Understanding these proteins' modes of action and functions in viral biology and disease is therefore of considerable importance, and the subject of this review.


The insulin gene is transcribed in the human thymus and transcription levels correlated with allelic variation at the INS VNTR-IDDM2 susceptibility locus for type 1 diabetes.
Type 1, or insulin-dependent diabetes mellitus (IDDM) is an autoimmune disease associated with loss of tolerance to several pancreatic islet cell molecules, including insulin, glutamic acid decarboxylase (GAD), ICA69 and the tyrosine phosphatase IA-2 (refs 1-3). Among several predisposing loci, IDDM2 maps to the insulin gene (INS) VNTR (variable number of tandem repeats) minisatellite on chromosome 11p15 (refs 4-9). Allelic variation at this VNTR locus correlates with steady-state levels of INS mRNA in pancreas and transfected rodent cell lines, but it is difficult to reconcile the association of lower INS mRNA levels in the pancreas with class III VNTRs that are dominantly protective from IDDM. We show that during fetal development and childhood, mRNAs for insulin and other islet cell autoantigens (GAD, ICA69, IA-2) are expressed at low levels in the human thymus. Critically, we also detect proinsulin and insulin protein. VNTR alleles correlate with differential INS mRNA expression in the thymus where, in contrast to the pancreas, protective class III VNTRs are associated with higher steady-state levels of INS mRNA expression. This finding provides a plausible explanation for the dominant protective effect of class III VNTRs, and suggests that diabetes susceptibility and resistance associated with IDDM2 may derive from the VNTR influence on INS transcription in the thymus. Higher levels of (pro)insulin in the thymus may promote negative selection (deletion) of insulin-specific T-lymphocytes which play a critical role in the pathogenesis of type-1 diabetes.


Are hemodynamic parameters predictors of mortality?
This article addresses a question that the authors consider to be somewhat rhetorical: are hemodynamic parameters predictors of mortality? It reviews the specific hemodynamic abnormalities and pathophysiologic consequences distinctive to the patient who has decompensation and addresses the data that implicate abnormal hemodynamics as a treatment target associated with increased mortality. The focus is on patients who have decompensated heart failure, defined as left ventricular systolic dysfunction and an acute, subacute, or gradual worsening of symptoms while receiving optimal medical therapy.


Clinical validation of high-resolution image reconstruction algorithms in brain 18F-FDG-PET: effect of incorporating Gaussian filter, point spread function, and time-of-flight.
Accurate estimation of radiopharmaceutical uptake in the brain is difficult because of count statistics, low spatial resolution, and smoothing filter. The aim of this study was to assess the counting rate performance of PET scanners and the image quality with different combinations of high-resolution image reconstruction algorithms in brain F-2-fluorodeoxy-D-glucose (F-FDG)-PET. Using 23 patient studies, we analyzed the coincidence rates of true and random, random fraction, and the noise equivalent counts per axial length (NECpatient) in brain and liver bed positions. The reconstruction algorithms were combined with baseline ordered subsets expectation maximization, Gaussian filter (GF), point spread function (PSF), and time-of-flight (TOF). The image quality of the brain cortex was quantitatively evaluated with respect to spatial resolution, contrast, and signal-to-noise ratio (SNR). The true coincidence rate in the brain was higher by 1.86 times and the random coincidence rate was lower by 0.61 times compared with that in the liver. In the brain, random fraction was lower and NECpatient was higher than that of the liver. Although GF improved the SNR, spatial resolution and contrast were reduced by 12 and 11%, respectively (P<0.01). PSF improved spatial resolution and SNR by 11 and 53%, respectively (P<0.01), and TOF improved SNR by ∼23% (P<0.01). We have demonstrated that a high-resolution image reconstruction algorithm for brain F-FDG-PET is promising without the use of a GF because of high true coincidence counts and that combined with PSF and TOF is optimal for obtaining a better SNR of the image.


Communication between the cell membrane and the nucleus: Role of protein compartmentalization.
Understanding how the information is conveyed from outside to inside the cell is a critical challenge for all biologists involved in signal transduction. The flow of information initiated by cell-cell and cell-extracellular matrix contacts is mediated by the formation of adhesion complexes involving multiple proteins. Inside adhesion complexes, connective membrane skeleton (CMS) proteins are signal transducers that bind to adhesion molecules, organize the cytoskeleton, and initiate biochemical cascades. Adhesion complex-mediated signal transduction ultimately directs the formation of supramolecular structures in the cell nucleus, as illustrated by the establishment of multi complexes of DNA-bound transcription factors, and the redistribution of nuclear structural proteins to form nuclear subdomains. Recently, several CMS proteins have been observed to travel to the cell nucleus, suggesting a distinctive role for these proteins in signal transduction. This review focuses on the nuclear translocation of structural signal transducers of the membrane skeleton and also extends our analysis to possible translocation of resident nuclear proteins to the membrane skeleton. This leads us to envision the communication between spatially distant cellular compartments (i.e., membrane skeleton and cell nucleus) as a bidirectional flow of information (a dynamic reciprocity) based on subtle multilevel structural and biochemical equilibria. At one level, it is mediated by the interaction between structural signal transducers and their binding partners, at another level it may be mediated by the balance and integration of signal transducers in different cellular compartments. J. Cell. Biochem. Suppls. 30/31:250-263, 1998. © 1998 Wiley-Liss, Inc.


A gene locus controlling a serum protein migrating electrophoretically in the beta region of mice and detected by using a strain derived from the Japanese wild mouse (Mus musculus molossinus).
Antigenic specificities of serum proteins from the MOL-ANJ strain of mice (a strain derived from Japanese wild mice, Mus musculus molossinus) were studied by gel precipitation with alloantisera produced by reciprocal alloimmunization between MOL-ANJ and BALB/c mice. An alloantigen which migrates immunoelectrophoretically in the beta region of serum proteins has been identified. Evidence indicates that this antigenic specificity is controlled by a co-dominant autosomal gene locus designated by the symbol Sas-2. The evidence also suggests that Sas-2 is genetically different from the previously described Sas-1 which controls a serum protein in the mouse. Sas-2 was located by linkage analysis between Idh-1 locus and Akp-1 locus on chromosome 1.