C/EBP alpha and beta mimic retinoic acid activation of IGFBP-5 in neuroblastoma cells by a mechanism independent from binding to their site.
Signal transduction mediated by insulin-like growth factors is implicated in the aggressive behavior of neuroblastoma (NB), a childhood tumor originating from the neural crest. IGFBP-5, a protein that binds IGFs with high affinity, is expressed in many NB cell lines exerting opposite effects, depending on its concentration. We found that IGFBP-5 expression increased during retinoic acid (RA)-mediated differentiation of NB cells. This was due to transcriptional activation as demonstrated by reporter assays carried out in basal and differentiating conditions. We defined the shortest region of the human IGFBP-5 promoter (from nucleotide -83 to +53) which is sensitive to RA. Mutation of a CCAAT enhancer binding protein (C/EBP) element inside this region increased transcription, suggesting a repressive role of this sequence. DNA Affinity Precipitation Assays (DAPA) and chromatin immunoprecipitation demonstrated that the binding of C/EBPalpha and beta to the C/EBP site decreased upon treatment with RA. C/EBPalpha and beta induced an increase in IGFBP-5 transcription in human and murine NB cells similar to that obtained upon RA treatment. Activation by C/EBP alpha and beta did not depend on their binding to the C/EBP site, since they still activated IGFBP-5 promoter carrying a mutation in the C/EBP site. Of interest, we found that both transcription factors were able to interact with the TATA box, but only C/EBPalpha interaction increased during RA-induced differentiation.


Impacting cost and appropriateness of stress ulcer prophylaxis at a university medical center.
To determine the appropriateness and medication cost of stress ulcer prophylaxis before and after a targeted educational intervention. In the preintervention cohort (phase 1), 264 patients were evaluated over 2 months, using stress ulcer prophylaxis guidelines developed by a comprehensive literature search. Targeted educational programs were subsequently used to inform trauma housestaff on appropriate usage of stress ulcer prophylaxis medications with emphasis on using sucralfate. The postintervention cohort (phase 2) involved concurrent evaluation of 279 patients. Length of inappropriate stress ulcer prophylaxis (i.e., did not meet approved guidelines) between phases was compared using a Student's t-test for independent samples (alpha = .05). A 365-bed university medical center. Patients admitted to any of the intensive care units and all patients who were placed on histamine-2-antagonists or sucralfate for stress ulcer prophylaxis. Educational intervention regarding appropriate stress ulcer prophylaxis directed at the trauma service. Patient demographics in the two phases were similar and there was no difference in the number of patient risk factors for stress-induced bleeding. The mean length of inappropriate stress ulcer prophylaxis was 5.78 +/- 4.36 days in phase 1 and 4.66 +/- 3.10 days in phase 2 (p < .05). Eighty-nine patients in phase 1 received inappropriate stress ulcer prophylaxis for a drug cost of $2,272.00 (mean $25.53 +/- 25.52) compared with 90 patients in phase 2 with a drug cost of $1,417.00 (mean $15.75 +/- 13.06). Three patients in each phase had clinically important bleeding (hemodynamic compromise or transfusion); all were receiving ranitidine. The mean total cost (fixed and variable) of hospitalization was $69,288.00 and $74,709.00 for the three patients who bled in each phase compared with $19,850.00 and $15,812.00 for all patients admitted to the intensive care unit in phases 1 and 2, respectively. The mean length of hospital stay was 30.00 days and 29.33 days for the three patients who bled in each phase compared with 11.54 days and 10.27 days for all patients admitted to the intensive care unit in phases 1 and 2, respectively. Cost savings are associated with more appropriate stress ulcer prophylaxis. Clinically important bleeding is uncommon but results in prolonged hospital stays and increased costs.


[The effect of anabasine on mortality and concentration of proinflammatory cytokines in blood of mice at early stage of sepsis].
It was established in experiments on noninbred mice that activation of α-7n acetylcholine receptors (α-7n AChR) by anabasine in single doses of 1.0 and 5.0 mg/kg for 2 h before modeling sepsis (intraperitoneal injection of E. coli) cause a significant dose-dependent reduction of mortality of mice due to a decrease in the amount of proinflammatory cytokines TNF-α, IL-1β, and IL-6 in the blood. Anabasine in single doses of 0.1 mg/kg had no significant impact on the studied parameters.


Dependence on antibody for induction of chemiluminescence in polymorphonuclear leukocytes by herpes simplex virus.
The induction of luminol-dependent chemiluminescence in rabbit polymorphonuclear leukocytes (PMN) by a stromal keratitis causing strain (RE) of herpes simplex virus type 1 (HSV-1) was examined. Virus alone and virus infected rabbit corneal cells were unable to stimulate chemiluminescence. However, when the virus or virus infected cells were incubated in the presence of HSV-1 specific immune serum or purified IgG, a gradual chemiluminescent response was observed. Virus and virus infected cells incubated with normal rabbit serum or IgG produced little or no activity. No impairment of chemiluminescent response was observed in experiments in which rabbit PMN were exposed to HSV prior to the addition of opsonized zymosan or HSV-antibody complexes. Results suggest PMN exert antiviral activity in the presence of specific antibody and may be important factors in the inflammatory process resulting from ocular HSV infection.


Analytical Profile and Antioxidant and Anti-Inflammatory Activities of the Enriched Polyphenol Fractions Isolated from Bergamot Fruit and Leave.
The aim of the study is to compare the qualitative and semi-quantitative profile of the polyphenol fraction purified from the leaf (BLPF) and fruit (BFPF) of bergamot (Citrus bergamia), and to evaluate their antioxidant and anti-inflammatory activity. The analytical qualitative profile was carried out by LC-ESI/MS using three different approaches: targeted (searching analytes already reported in bergamot extract), semi-targeted (a selective search of 3-hydroxy-3-methylglutarate [HMG] derivatives involved in the cholesterol reducing activity of BPF) and untargeted. A total number of 108 compounds were identified by using the three approaches, 100 of which are present in both the extracts thus demonstrating a good qualitative overlapping of polyphenols between the two extracts. The antioxidant activity was higher for BLPF in respect to BFPF but when normalized in respect to the polyphenol content they were almost overlapping. Both the extracts were found to dose dependently inhibit cell inflammation stimulated with IL-1α. In conclusion, the comparison of the qualitative and quantitative profile of polyphenols as well as of the antioxidant and anti-inflammatory activity of bergamot leaf and fruit well indicates that leaf is a valid source of bergamot polyphenol extraction and an even richer source of polyphenol in respect to the fruit.


MSH-producing gastric tumour.
MSH-producing gastric tumour. A case report. A patient with melanosis due to MSH-production by a gastric tumour is described. It is also possible that the tumour was producing gastrin, whereas there was no sign of increased ACTH-production. This is the first patient described with a MSH-producing tumour without concomitant ACTH-production.


Does public report card matter? A 10-year interrupted time series analysis on total knee replacement.
There is a lack of evidence that shows whether a report card can improve health outcomes in terms of infection rates or unscheduled readmission by using rigorous methods to evaluate its impact. We used the National Health Insurance Administration's claims database from 1 January 2004 to 30 December 2013 and a time series analysis to evaluate the impact of the quality report card initiative on three negative outcomes of total knee replacement for each quarter of the year, including the rates of superficial infection of a knee replacement, deep infection of knee arthroplasty and unplanned readmissions for surgical site infection. These negative outcomes (original scale) do not show significant decreases in terms of superficial infection (-0.05‰, -0.63 to 0.53‰, P = 0.87), deep infection (-0.003‰, -0.19 to 0.18‰, P = 0.97) and unscheduled readmission (0.02‰, -0.21 to 0.25‰, P = 0.88). The total knee replacement public report card initiative did not improve the rate of infection and unscheduled readmission for surgical site infection. This report card in Taiwan should involve physicians' participation in the design and be tailored to be suitable for reading by patients in order to further enhance the chance of improvement in these negative outcomes.


[Air humidity solubility assay].
Measurement of drug solubility is one of the key elements of compound characterization during the drug discovery and development process. A broad variety of solubility assay methods have been developed, including equilibrium method which requires analysis of the equilibrium composition and kinetic method which monitors the concentration of a compound dynamically at the time when a precipitate first appears or disappears in the solution. Despite the numerous experimental methods, precise drug solubility values are hard to obtain for time-consuming, sample size and manual work. In this article, we reported a new method, namely air humidity solubility assay, which measures the relative humidity of the air in equilibrium with the solution at a given temperature, and then calculates solubility from the relative humidity according to extended-non random two liquid (NRTL) model. NaCl was used as a model drug, and the solubility was measured at the temperature of 20-50 ℃. The results indicate that the solubility of NaCl determined with the new method is generally comparable to that determined by gravimetry that is reported in literature. The new method has a relative error of less than 2%. Although the accuracy is lower than that of gravimetry, air humidity solubility assay is more convenient, practical, operational and universal. This method provides a supplement to the existing methods.


The dexamethasone/corticotropin-releasing factor test in men with major depression: role of childhood trauma.
The dexamethasone/corticotropin-releasing factor (CRF) test is considered to be the most sensitive measure of hypothalamic-pituitary-adrenal (HPA) axis hyperactivity and has been demonstrated to be altered in patients with major depression (MDD). Although childhood trauma is a demonstrated risk factor for MDD and patients with a history of childhood abuse and MDD demonstrate HPA axis hyperactivity, the dexamethasone/CRF test remains unstudied in this population. We determined the impact of childhood trauma on dexamethasone/CRF test results in patients with MDD. Forty-nine healthy men, ages 18-60 years, without mania or psychosis, active substance abuse, or eating disorder and medication-free were recruited into four study groups, including: 1) normal subjects with no childhood abuse history or psychiatric disorder (n = 14); 2) men with childhood abuse histories without current MDD (n = 14); 3) men with childhood abuse histories with current MDD (n = 15); and 4) men with current MDD and no childhood abuse history (n = 6). Plasma adrenocorticotropin (ACTH) and cortisol concentrations were measured in response to dexamethasone/CRF administration. Men with childhood trauma histories exhibited increases in ACTH and cortisol responses to dexamethasone/CRF compared with non-abused men. In particular, abused men with current MDD showed increased responsiveness compared with control subjects and depressed men without childhood abuse experience. Increased response was associated with the severity, duration, and earlier onset of the abuse. The effects were not explained by concurrent posttraumatic stress disorder. Childhood trauma increases HPA axis activity as measured with the dexamethasone/CRF test in adult men with MDD, potentially reflecting environmental risk for developing depression.


[Hypothesis: phagocytosis of aberrant cells protects long life span vertebrate species against tumors].
Probable mechanisms of resistance to carcinogenic impacts and spontaneous tumours of long life span vertebrate species have been discussed. It has been suggested that this compatible with long life span mechanism is likely phagocyte elimination with involving DNA-protenkinase-dependent, wt p53 or/and wt p53-dependent signaling pathways also ligands for "Scavenger receptor" and "Toll-like receptors". The approaches to the experimental evidence of this hypothesis have been produced.


Feeding frequency does not interact with BPA exposure to influence metabolism or behaviour in zebrafish (Danio rerio).
Resource limitation can constrain energy (ATP) production, and thereby affect locomotion and behaviour such as exploration of novel environments and boldness. Consequently, ecological processes such as dispersal and interactions within and between species may be influenced by food availability. Energy metabolism, and behaviour are regulated by endocrine signalling, and may therefore be impacted by endocrine disrupting compounds (EDCs) including bisphenol A (BPA) derived from plastic manufacture and pollution. It is important to determine the impacts of these novel environmental contexts to understand how human activity alters individual physiology and behaviour and thereby populations. Our aim was to determine whether BPA exposure interacts with feeding frequency to alter metabolism and behaviour. In a fully factorial experiment, we show that low feeding frequency reduced zebrafish (Danio rerio) mass, condition, resting metabolic rates, total distance moved and speed in a novel arena, as well as anxiety indicated by the number of times fish returned to a dark shelter. However, feeding frequency did not significantly affect maximal metabolic rates, aerobic scope, swimming performance, latency to leave a shelter, or metabolic enzyme activities (citrate synthase and lactate dehydrogenase). Natural or anthropogenic fluctuation in food resources can therefore impact energetics and movement of animals with repercussions for ecological processes such as dispersal. BPA exposure reduced LDH activity and body mass, but did not interact with feeding frequency. Hence, behaviour of adult fish is relatively insensitive to disruption by BPA. However, alteration of LDH activity by BPA could disrupt lactate metabolism and signalling and together with reduction in body mass could affect size-dependent reproductive output. BPA released by plastic manufacture and pollution can thereby impact conservation and management of natural resources.


Versatile chelating behavior of benzil bis(thiosemicarbazone) in zinc, cadmium, and nickel complexes.
Reactions of benzil bis(thiosemicarbazone), LH(6), with M(NO(3))(2).nH(2)O (M = Zn, Cd, and Ni), in the presence of LiOH.H(2)O, show the versatile behavior of this molecule. The structure of the ligand, with the thiosemicarbazone moieties on opposite sides of the carbon backbone, changes to form complexes by acting as a chelating molecule. Complexes of these metal ions with empirical formula [MLH(4)] were obtained, although they show different molecular structures depending on their coordinating preferences. The zinc complex is the first example of a crystalline coordination polymer in which a bis(thiosemicarbazone) acts as bridging ligand, through a nitrogen atom, giving a 1D polymeric structure. The coordination sphere is formed by the imine nitrogen and sulfur atoms, and the remaining position, in a square-based pyramid, is occupied by an amine group of another ligand. The cadmium derivative shows the same geometry around the metal ion but consists of a dinuclear structure with sulfur atoms acting as a bridge between the metal ions. However, in the nickel complex LH(6) acts as a N(2)S(2) ligand yielding a planar structure for the nickel atom. The ligand and its complexes have been characterized by X-ray crystallography, microanalysis, mass spectrometry, IR, (1)H, and (13)C NMR spectroscopies and for the cadmium complex by (113)Cd NMR in solution and in the solid state.


Sesquiterpene lactones downregulate G2/M cell cycle regulator proteins and affect the invasive potential of human soft tissue sarcoma cells.
Soft tissue sarcomas (STS) represent a rare group of malignant tumors that frequently exhibit chemotherapeutic resistance and increased metastatic potential. Many studies have demonstrated the great potential of plant-derived agents in the treatment of various malignant entities. The present study investigates the effects of the sesquiterpene lactones costunolide and dehydrocostus lactone on cell cycle, MMP expression, and invasive potential of three human STS cell lines of various origins. Both compounds reduced cell proliferation in a time- and dose-dependent manner. Dehydrocostus lactone significantly inhibited cell proliferation, arrested the cells at the G2/M interface and caused a decrease in the expression of the cyclin-dependent kinase CDK2 and the cyclin-dependent kinase inhibitor p27(Kip1). In addition, accumulation of cells at the G2/M phase transition interface resulted in a significant decrease in cdc2 (CDK1) together with cyclin B1. Costunolide had no effect on the cell cycle. Based on the fact that STS tend to form daughter cell nests and metastasize, the expression levels of matrix metalloproteinases (MMPs), which play a crucial role in extracellular matrix degradation and metastasis, were investigated by Luminex® technology and real-time RT-PCR. In the presence of costunolide, MMP-2 and -9 levels were significantly increased in SW-982 and TE-671 cells. Dehydrocostus lactone treatment significantly reduced MMP-2 and -9 expression in TE-671 cells, but increased MMP-9 level in SW-982 cells. In addition, the invasion potential was significantly reduced after treatment with both sesquiterpene lactones as investigated by the HTS FluoroBlock™ insert system.


ZnO quantum dots modified bioactive glass nanoparticles with pH-sensitive release of Zn ions, fluorescence, antibacterial and osteogenic properties.
Zinc (Zn)-containing materials have osteogenic and antibacterial activities while bioactive glass nanoparticles (BGN) show bone-bonding ability, as well as osteoconductive and osteoinductive properties. Zn-containing BGN are therefore considered to be promising materials for various biomedical applications, particularly in bone regeneration. In this study, we report a convenient method to prepare Zn-containing BGN by coating ZnO quantum dots (QDs) on BGN via electrostatic interactions. The synthesized ZnO-BGN nanocomposite particles are spherical and highly dispersed, and exhibit a unique fluorescence behavior under UV excitation, emitting three wavelengths in the violet, blue and green range. ZnO-BGN showed apatite-forming ability upon immersion in simulated body fluid, but their apatite formation was delayed compared to BGN. Interestingly, ZnO-BGN showed a rapid release of Zn ions at pH 4 but a far slower release at pH 7.4. ZnO-BGN also exhibited antibacterial effects on both Gram-positive and Gram-negative bacteria at the concentrations of 1, 0.1, and 0.01 mg mL-1. Higher concentrations could lead to stronger antibacterial effects. The LDH and live/dead assays indicated that ZnO-BGN had no significant cytotoxicity towards human mesenchymal stem cells (hMSC) at concentration of 0.1 and 0.01 mg mL-1, but ZnO-BGN inhibited the relative proliferation of hMSC compared to BGN and the control according to the MTT assay. Notably ZnO-BGN improved the osteogenic differentiation of hMSC as indicated by the determination of the alkaline phosphatase activity. In conclusion, coating quantum dots on BGN is a promising strategy to produce Zn-containing BGN. The synthesized ZnO-BGN are potential materials for bone regeneration, considering their apatite-forming ability, unique ion-release behavior, effective antibacterial activity, non-cytotoxicity, and osteogenic potential.


Development of a Brazilian Portuguese version of the Oswestry Disability Index: cross-cultural adaptation, reliability, and validity.
Translation, revision, back-translation, and 3-way validity were performed. The objective of this study was to translate a version of the Oswestry Disability Index (ODI) into Brazilian Portuguese and evaluate its reliability. Reports in the literature have identified a need for internationally standardized and reliable measurements to analyze back pain. The ODI has become one of the principal outcome measurements used in the management of spinal disorders. The cross-cultural adaptation was performed according to the internationally recommended methodology, using the following guidelines: translation, back-translation; revision by a committee, and pretesting. The psychometric properties were evaluated by administering the questionnaire to 120 subjects with back pain. Reliability was estimated through stability and homogeneity assessment. The validity was tested comparing ODI scores with the following measurements: Roland-Morris Disability Questionnaire, SF-36, and a Numerical Pain Scale. Good internal consistency was found (Cronbach's alpha = 0.87). Intraclass correlation coefficient for test-retest reliability was 0.99. The ODI showed moderate correlation with pain measurement (r = 0.66). Relatively high correlation was also found between the ODI and the Roland-Morris scores (r = 0.81). There was significant correlation (P < 0.001) between ODI scores and the 8 scales of the SF-36. The data showed that the cultural adaptation process was successful and that the adapted instrument demonstrated having excellent psychometric properties, reliable in the Brazilian culture.


A Systematic Review and Meta-analysis of the Effort-Reward Imbalance Model of Workplace Stress and Hypothalamic-Pituitary-Adrenal Axis Measures of Stress.
The association between effort-reward imbalance (ERI) and various health outcomes has been well documented over the past 20 years, but the mechanisms responsible for this association remain unclear. The present meta-analysis assessed the associations of ERI and overcommitment (OC) in the workplace with hypothalamic-pituitary-adrenal (HPA) axis measures. Electronic databases were searched with the phrase "effort*reward*imbalance," which yielded 319 studies leading to 56 full-text studies being screened. Thirty-two studies within 14 articles met inclusion criteria and were meta-analyzed using mixed and random effects models. Greater ERI was associated with increased HPA axis activity (r = .09, p < .001, k = 14, N = 2541). The cortisol awakening response (r = .14, p < .001, k = 9, N = 584) and cortisol waking concentrations (r = .12, p = .01, k = 6, N = 493) were the only HPA measures associated with ERI. OC was also associated with greater HPA axis activity (r = .06, p < .01, k = 10, N = 1918). Cortisol (PM) (r = .13, p = .02, k = 3, N = 295) was the only HPA measure associated with OC. ERI and OC were similarly related with HPA responsivity. However, because OC moderated the relationship between ERI and HPA axis markers, the importance of OC should not be overlooked. Because OC is likely more malleable than ERI to intervention, this may be a promising avenue for future research.


Robust scalable synthesis of a bis-urea derivative forming thixotropic and cytocompatible supramolecular hydrogels.
Synthetic hydrogels address a need for affordable, industrially scalable scaffolds for tissue engineering. Herein, a novel low molecular weight gelator is reported that forms self-healing supramolecular hydrogels. Its robust synthesis can be performed in a solvent-free manner using ball milling. Strikingly, encapsulated cells spread and proliferate without specific cell adhesion ligands in the nanofibrous material.


Seasonal variations in the histology of the pituitary gland of Cirrhinus mrigala (Ham.) an Indian freshwater major carp, in relation to gonadal activity.
1. Seasonal variations in the histology of the pituitary gland of Cirrhinus mrigala (Ham.), an Indian freshwater major carp, in relation to gonadal activity has been studied during a one year period commencing, January, 1971. 2. The proximal pars distalis undergoes a great deal of seasonal variation in size, weight and nature of pituicytes. 3. The percentage composition of acidophils and cyanophils in the proximal pars distalis show a reciprocal relationship during the year. The acidophils predominate during the restitution phase of the gonads while the cyanophils preponderate among the different cell types during the breeding season. 4. The chromophobes do not show any significant fluctuation in numerical abundance in relation to gonadal activity. 5. Cyanophils of the proximal pars distalis undergo changes with change in gonadal activity Concentration of glycoproteinaceous material is extremely low during the restitution phase but high during the maturation phase. The cyanophils also undergo hypertrophy and hyperplasia during the spawning period. The latter discharge their contents and become vacuolated during the post spawning period. 6. The number and size of the globules in the cyanophils are related to gonadal activity. 7. Acidophils present in the rostral pars distalis and pars intermedia do not appear to be related to gonadal activity in C. mrigala. 8. The average weight of the gland is directly related to the state of maturation of the gonads, an observation being recorded here for the first time.


Radiation combines with immune checkpoint blockade to enhance T cell priming in a murine model of poorly immunogenic pancreatic cancer.
Radiation has been a pillar of cancer therapy for decades. The effects of radiation on the anti-tumour immune response are variable across studies and have not been explicitly defined in poorly immunogenic tumour types. Here, we employed combination checkpoint blockade immunotherapy with stereotactic body radiation therapy and examined the effect on tumour growth and immune infiltrates in subcutaneous and orthotopic mouse models of pancreatic cancer. Although immune checkpoint blockade and radiation were ineffective alone, their combination produced a modest growth delay in both irradiated and non-irradiated tumours that corresponded with significant increases in CD8+ T cells, CD4+ T cells and tumour-specific T cells as identified by IFNγ ELISpot. We conclude that radiation enhances priming of tumour-specific T cells in poorly immunogenic tumours and that the frequency of these T cells can be further increased by combination with immune checkpoint blockade.


The association between childhood trauma and lifetime suicide attempts among a sample of male prisoners: A pilot observational study.
Prisoners have higher rates of suicide attempts compared with general population. A history of childhood trauma (CT) is common among incarcerated subjects and it is a well-known risk factor for lifetime suicide attempts. Therefore, the purpose of the study was to investigate whether lifetime suicide attempts may be related to the exposition to CT among male prisoners. We conducted a cross sectional study recruiting newly arrived inmates in an Italian jail, between January 2017 and June 2018. Prisoners were interviewed to collect socio-demographic and clinical information. Moreover, inmates completed the Childhood Trauma Questionnaire. We excluded prisoners unable to speak or read Italian, with learning disabilities or current severe psychiatric symptoms. A total of 215 consecutive male inmates were included. Fifty-one prisoners (23.7%) had a history of attempted suicide. The most reported CT was physical neglect. Multivariate logistic regression analysis showed that a history of childhood sexual abuse, emotional neglect and psychiatric diagnosis significantly increased the likelihood of lifetime suicide attempt. A previous history of suicide attempt is highly prevalent among inmates. In agreement with previous findings, lifetime suicide attempts seem to be associated with the presence of CT and psychiatric diagnosis. Therefore, CT should be considered as a relevant variable to improve the programs for the prevention of suicide in prison.


Making sense of how physician preceptors interact with medical students: discourses of dialogue, good medical practice, and relationship trajectories.
Work based learning and teaching in health care settings are complex and dynamic. Sociocultural theory addresses this complexity by focusing on interaction between learners, teachers, and their environment as learners develop their professional identity. Although social interaction between doctors and students plays a crucial role in this developmental process, socio-cultural research from the perspective of doctors is scarce. We performed discourse analysis on seven general practitioners' audio diaries during a 10-week general practice clerkship to study how they gave shape to their interaction with their students. Examination of 61 diary-entries revealed trajectories of developing relationships. These trajectories were initiated by the way respondents established a point of departure, based on their first impression of the students. It continued through the development of dialogue with their student and through conceptualizations of good medical practice. Such conceptualizations about what was normal in medical and educational practice enabled respondents to recognize qualities in the student and to indirectly determine students' desired learning trajectory. Towards the end, discursive turns in respondents' narratives signaled development within the relationship. This became evident in division of roles and positions in the context of daily practice. Although respondents held power in the relationships, we found that their actions depended strongly on what the students afforded them socially. Our findings address a gap in literature and could further inform theory and practice, for example by finding out how to foster constructive dialogue between doctors and students, or by exploring different discourses among learners and teachers in other contexts.


Characteristics and Outcomes of Patients with Hematological Malignancies Admitted for Intensive Care - a Single Centre Experience
Background and Purpose: Patients with hematological malignancies admitted to an intensive care unit (ICU) generally have a high mortality rate. The aim of our study was to assess the characteristics and outcomes of such patients and to identify factors predicting ICU mortality. Material and Methods: This retrospective chart review was conducted in the intensive care unit (ICU) of Shaukat Khanum Memorial Cancer Hospital and Research Centre over a period of 5 years, from January 2010 to January 2015. Results: Characteristics :A total of 213 patients were included in this study. There were 150 (70.4%) males and 63 (29.6%) females with the median age of 36 years (18-88 years). Main diagnosis was non- Hodgkin lymphoma in 127 (59.6%) followed by Hodgkin's disease in 27 (12.7%) and acute myeloid leukemia in 16 (7.5%). Most of the patients 154 (72.3%) were on active chemotherapy at the time of admission to ICU, while 28 patients (13.1%) had newly diagnosed disease and 22 (10.3%) featured either relapsed or progressive disease. The most common reason for admission to ICU was a combination of respiratory failure with septic shock (29.6%) followed by septic shock alone (19.7%) and acute respiratory failure (13.1%). Other causes included acute renal failure, alone (7.5%) or in combination with respiratory or circulatory collapse (10.8%) and central nervous system involvement (5.6%). The majority of admissions to ICU occurred between days one and five of admission to a ward (46.5%, n=99) whereas 49 (23%) were taken directly to the ICU. Mainstay of treatment in 38.5% of patients included both invasive ventilation and vasopressor support along with other supportive care like fluids and antibiotics. 23.5% received only supportive management. Duration of stay for 150 (70.4%) patients was between one to seven days. Outcomes: A total of 119 (55.9 %) patients expired while in ICU, while 14 (6.6%) died in hospital after being transferred out of ICU. ICU survival was 44.1% whereas hospital survival was 37.5%. After discharge from hospital in a stable condition, 18 (8.5%) patients were lost to follow up and 62 (29%) patients were alive after thirty days. A total of 33 (15.4 %) of patients survived for at least one year after ICU admission. Some 21 (9.8%) are still alive and healthy after a minimum median follow up of one and a half years. Predictors of Mortality: Overall, mechanical ventilation was required in 61% of patients. Out of the patients who expired, 92.4% required intubation, in contrast to 21.3% for those who survived the ICU stay. Involvement of three or more organs was apparent in 12.8% of improved patients and 70.6% of those who died during ICU stay. Neutropenia did not appear to be a major discriminatory factor, with 33% of improved and 42.9% of expired patients being neutropenic at the time of admission to ICU. The majority of patients from both the improved and expired group required intubation and vasopressors from day one onwards. Conclusions: Admission of patients with hematological malignancies to the intensive care unit is associated with poor outcome and high mortality. Identifying the patients who can benefit from aggressive care and prolonged ICU support is important especially when it comes to countries like ours with limited resources and major financial restraints. Multi-organ damage and requirement of invasive ventilation are two main predictors of increased mortality. Neutropenia is also associated with adverse outcome; however, the difference is not as significant as for the other two factors.


Hamstring Injuries: Critical Analysis Review of Current Nonoperative Treatments.
Hamstring injuries are common among active and athletic individuals, especially those involved in high-speed running, distance running, or sports requiring sudden directional changes. Acute hamstring strains often occur as an eccentric strain during running or a stretch-type injury caused by simultaneous hip flexion and knee extension. Proximal hamstring tendinopathy is an overuse injury of the hamstring tendon as a result of chronic cicatrization of the musculotendinous unit. Repeated stress to the hamstring tendon leads to increased cellularity of tendon fibers, disruption of collagen, and subsequent microinjury of the tissue that attaches the tendon to bone. Management of hamstring injuries generally begins with nonoperative modalities consisting of eccentric rehabilitative exercise programs. Although various other treatment modalities are available, the comparative efficacy of these supportive measures is not well differentiated at this time. In this article, we review the current literature with regard to the nonoperative treatment of hamstring injuries, specifically focusing on acute hamstring muscle strains and proximal hamstring tendinopathy in order to provide supplementary insight on the effectiveness of current modalities.


Impact of OATP1B1, MDR1, and CYP3A4 expression in liver and intestine on interpatient pharmacokinetic variability of atorvastatin in obese subjects.
Individual variability in expression and function of organic anion-transporting polypeptide 1B1 (OATP1B1), multidrug resistance protein 1 (MDR1), and/or cytochrome P450 3A4 (CYP3A4) may impact the clinical response of many drugs. We investigated the correlation between expression of these proteins and pharmacokinetics of atorvastatin, a substrate of all three, in 21 obese patients with paired biopsies from liver and intestinal segments. The patients were also screened for the SLCO1B1 c.521T→C variant alleles. Approximately 30% (r(2) = 0.28) of the variation in oral clearance (CL/F) of atorvastatin was explained by hepatic OATP1B1 protein expression (P = 0.041). Patients carrying the SLCO1B1 c.521C variant allele (homozygous, n = 4; heterozygous, n = 2) exhibited 45% lower CL/F of atorvastatin than the c.521TT carriers (P = 0.067). No association between hepatic and intestinal expression of MDR1 or CYP3A4 and atorvastatin pharmacokinetics was found (P > 0.149). In conclusion, this study suggests that OATP1B1 phenotype is more important than CYP3A4 and MDR1 phenotypes for the individual pharmacokinetic variability of atorvastatin.


[Prehistory of Parkinson's disease].
Parkinson's disease (PD) was described according to conventional history in 1817. However, PD has been known and partially described since the Biblical times. There are descriptions of parkinsonian symptoms in Indian and Egyptian texts. Galen also paid attention to rest tremor. Parkinsonian symptoms were also mentioned by several authors since the XVII century including Tulp, Silvio de la Boe, Johannes Baptiste Sagar, Gaubius, Boissier de Sauvages, Hunter and Chomel. Certainly James Parkinson deserves to be recalled after his text devoted to shaking palsy, but he also recognized he contribution of previous authors.


The role of vascular laboratory criteria in the selection of patients for lower extremity amputation.
We evaluated clinical and vascular laboratory data on 126 patients with below-knee or forefoot amputation. Vascular laboratory examination included Doppler systolic blood pressure and arterial wave form analysis using the segmental plethysmograph. Fifty-four patients had below-knee amputation. A calf systolic pressure greater than 70 torr was associated with 97% (33/34) success (p less than 0.005), an ankle systolic pressure greater than 30 torr yielded 91% (39/43) success (p less than 0.025), and an ankle systolic pressure greater than zero yielded an 87% success (p less than 0.005). In the absence of each of the above criteria, the predictive value of a negative test was only 32%, 40%, and 52%, respectively. The presence of a popliteal pulse was associated with 97% success (p less than 0.025); however, 88% of those with an absent popliteal pulse also achieved successful healing of below-the-knee amputations. Prior vascular reconstructive surgery was detrimental to healing of below-knee amputations. with 33% failure rate (p less than 0.025). For the 72 forefoot amputations, an ankle systolic pressure greater than 70 torr yielded a 65% success (p less than 0.025). The sensitivity of an ankle systolic pressure greater than 70 torr was 80% (32/40) and an ankle systolic greater than 35 yielded a sensitivity of 95% (38/40). The specificity was low for both of these reference values. Clinical and vascular laboratory criteria can identify patients who will have a successful below-knee amputation; however, because of the high false negative rate, patients should not be denied below-knee amputation solely on the basis of Doppler systolic pressure. Vascular laboratory criteria for predicting healing of forefoot amputations are also limited by the high rate of false positive and false negative results.


Quality of life and patient satisfaction in patients with atrial fibrillation on stable vitamin K antagonist treatment or switched to a non-vitamin K antagonist oral anticoagulant during a 1-year follow-up: A PREFER in AF Registry substudy.
Non-vitamin K antagonist oral anticoagulants (NOACs) are being introduced for stroke prevention in non-valvular Atrial Fibrillation (AF), and promise to be accepted better than Vitamin K Antagonists (VKAs) by patients, improving their Quality of Life (QoL). To assess to what extent patient-related factors influence decisions to switch from a VKA to a NOAC. The PREFER in AF Registry collected data at baseline in 2012 - at the beginning of NOAC prescriptions - and at 1-year follow-up, in 6412 patients in seven Western European countries. QoL and patient satisfaction questionnaires (EQ-5D-5L and/or PACT-Q2) were completed in 3777 patients at both visits. Data were compared across categories of patients on stable treatment with a VKA (i.e. continuously over the previous 12 months) (n=2102) or recently switched (within 12 months) from a VKA to a NOAC (n=213) during a 1-year follow-up, allowing a snapshot of factors influencing the switch at a time when NOACs were being introduced into the market. Compared to patients on stable treatment with a VKA, switched patients were similar in terms of age, sex, body mass index and other risk factors, but had lower prevalences of hypertension and heart valve dysfunction, and a lower rate of use of concomitant treatment with antiplatelet/anti-inflammatory agents; they also had a lower CHA2DS2-VASc score. Among 25 features investigated, switched patients more often reported bruising or bleeding, complained about bruising, were dissatisfied with the anticoagulant treatment, and reported mobility problems and anxiety/depressive traits. At the beginning of NOAC prescriptions, European doctors tended to switch from VKAs to NOACs those patients at lower risk than "non-switchers". Complaints about bruising or bleeding, dissatisfaction with treatment, mobility problems and anxiety/depression traits appear to be related to - and may have influenced - the choice to switch from a VKA to a NOAC.


Far Red-Shifted CdTe Quantum Dots for Multicolour Stimulated Emission Depletion Nanoscopy.
Stimulated emission depletion (STED) nanoscopy is a widely used nanoscopy technique. Two-colour STED imaging in fixed and living cells is standardised today utilising both fluorescent dyes and fluorescent proteins. Solutions to image additional colours have been demonstrated using spectral unmixing, photobleaching steps, or long-Stokes-shift dyes. However, these approaches often compromise speed, spatial resolution, and image quality, and increase complexity. Here, we present multicolour STED nanoscopy with far red-shifted semiconductor CdTe quantum dots (QDs). STED imaging of the QDs is optimized to minimize blinking effects and maximize the number of detected photons. The far-red and compact emission spectra of the investigated QDs free spectral space for the simultaneous use of fluorescent dyes, enabling straightforward three-colour STED imaging with a single depletion beam. We use our method to study the internalization of QDs in cells, opening up the way for future super-resolution studies of particle uptake and internalization.


Gibbon ape leukaemia virus RNA in leukaemic T-lymphoid cell lines: expression of a novel RNA transcript.
Fibroblast cell lines infected in vitro with different strains of gibbon ape leukaemia virus or the related woolly monkey virus (SSAV) synthesized two RNA species of approximately 8.4 kb and 2.9 kb. The former, a complete RNA, represents the gag-pol mRNA, while the latter is a spliced transcript lacking gag and pol, and represents the env mRNA. In contrast, RNA from one T-lymphoid cell line derived from a gibbon ape T-lymphocytic leukaemia (UCD-144) expressed a viral mRNA in addition to gag-pol and env mRNA. This RNA is 6.4 kb and lacks at least 3.0 kb of sequences derived from the internal region of the viral genome, including most or all of the pol gene. These data, as well as data from Southern blots of UCD-144 DNA, suggest that the 6.4 kb mRNA could represent a transcript from a defective recombinant provirus and may contain cell-derived sequences.


Diffuse-illumination holographic optical coherence tomography.
Holographic optical coherence tomography (OCT) is a powerful imaging technique, but its ability to reveal low-reflectivity features is limited. In this study, we performed holographic OCT by incoherently averaging volumes with changing diffuse illumination of numerical aperture (NA) equal to the detection NA. While the reduction of speckle from singly scattered light is only modest, we discovered that speckle from multiply scattered light can be arbitrarily reduced, resulting in substantial improvements in image quality. This technique also offers the advantage of suppressing noises arising from spatial coherence, and can be implemented with a partially spatially incoherent light source for further mitigation of multiple scattering. Finally, we show that although holographic reconstruction capabilities are increasingly lost with decreasing spatial coherence, they can be retained over an axial range sufficient to standard OCT applications.


Targeted delivery of anticancer drugs to tumor vessels by use of liposomes modified with a peptide identified by phage biopanning with human endothelial progenitor cells.
As tumor angiogenic vessels are critical for tumor growth and express different molecules on their surface from those on normal vessels, these vessels are expected to be an ideal target for anticancer drug delivery systems. It was previously reported that endothelial progenitor cells (EPCs) are involved in angiogenesis, tumor growth, and metastasis, and that EPCs show gene expression patterns similar to those of tumor endothelial cells. In the present study, a tumor vessel-targeting peptide, ASSHN, was identified from a phage-display peptide library by in vitro biopanning with human EPCs (hEPCs) and in vivo biopanning using angiogenesis model mice prepared by the dorsal air sac method. Phage clones displaying ASSHN peptide showed a marked affinity for hEPCs in vitro, and also for tumor vessels in vivo. PEGylated liposomes modified with the ASSHN peptide (ASSHN-Lip) were designed and prepared for the delivery of anticancer agents. Confocal images showed that ASSHN-Lip clearly bound to hEPCs in vitro and tumor vessels, and also showed extravasation from the vessels. The administration of doxorubicin-encapsulated ASSHN-Lip into Colon26 NL-17-bearing mice significantly suppressed tumor growth compared with doxorubicin-encapsulated PEGylated liposomes. These results suggest that the delivery of anticancer agents with ASSHN-Lip could be useful for targeted cancer therapy.


Protective effects of hydroxyethyl starch-deferoxamine in early sepsis.
The protective effects of hydroxyethyl starch-conjugated deferoxamine (HES-DFO), a macromolecular iron chelator, on the initial pathophysiological cascade in septic shock were evaluated following cecal ligation puncture (CLP) in rats. Animals were given an intravenous dose of 3.0 mL of either vehicle (HES) or HES-DFO immediately following completion of the CLP procedure. Animals were sacrificed 30, 60, 120, and 240 min following CLP, and samples of lung, kidney, bowel, and liver were collected for subsequent analysis of glutathione, myeloperoxidase, and evidence for lipid peroxidation based on measurement of thiobarbituric acid reactive substances and conjugated dienes. In addition, the endotoxin levels were determined in the plasma and histomorphological examination was conducted on tissue samples collected at each time point. At almost all time points, a reduction in lipid peroxidation was noted in the HES-DFO-treated rats (p < .05). Glutathione and myoloperoxidase levels were less affected. Lung tissue from animals receiving HEs demonstrated marked microatelectases, septal destruction, and splicing of basal membranes, which were greatly attenuated in animals having received HES-DFO. Similarly, tubulotoxic and mitochondrial damages observed in kidney samples from HES-treated animals were noticeably reduced in the animals having received the chelator. Liver and gut samples demonstrated unspecific inflammatory injury in both groups of animals. In summary, oxygen radical-mediated tissue damage occurs rapidly following CLP-induced sepsis. Based on histological and biochemical endpoints, treatment with the polymeric iron chelator, HES-DFO, significantly attenuates systemic oxidant injury, the degree of protection being most impressive in the lung and kidney.


Human babesiosis--an unrecorded reality. Absence of formal registry undermines its detection, diagnosis and treatment, suggesting need for immediate mandatory reporting.
Human babesiosis, caused by parasitic protozoa of erythrocytes, has escaped usual associates--lower mammals. Thriving in tick guts, it has spread inland from the coasts of America, adopting mankind as a host. Babesia spp. threaten life quality of unsuspecting humans in quickly expanding territories worldwide, including the state of Pennsylvania, USA. The causative spirochetes of Lyme disease often similarly co-exist in ticks. Singly or together they may, by causing persistent and chronic infections, duplicate any symptom in the medical literature--including depression and hypochondriasis. Physicians practicing throughout Pennsylvania have identified patients with symptomatic babesiosis, but without governmental surveillance or health registries that require doctors to consider and report babesiosis, these cases have not prompted epidemiological concern. Misunderstandings such as, "Isn't that an obscure tropical disease?" are usual responses when doctors are asked about babesiosis, inadvertently trivializing patients and disease. Mandatory reporting of babesiosis should now be considered a medical necessity.


Union Transitions Following the Birth of a Child to Cohabiting Parents.
Despite a growing interest in the family trajectories of unmarried women, there has been limited research on union transitions among cohabiting parents. Using data from the 2002 National Survey of Family Growth, we examined how family complexity (including relationship and fertility histories), as well as characteristics of the union and birth, were associated with transitions to marriage or to separation among 1,105 women who had a birth in a cohabiting relationship. Cohabiting parents had complex relationship and fertility histories, which were tied to union transitions. Having a previous nonmarital birth was associated with a lower relative risk of marriage and a greater risk of separation. In contrast, a prior marriage or marital birth was linked to union stability (getting married or remaining cohabiting). Characteristics of the union and birth were also important. Important racial/ethnic differences emerged in the analyses. Black parents had the most complex family histories and the lowest relative risk of transitioning to marriage. Stable cohabitations were more common among Hispanic mothers, and measures of family complexity were particularly important to their relative risk of marriage. White mothers who began cohabiting after conception were the most likely to marry, suggesting that ''shot-gun cohabitations'' serve as a stepping-stone to marriage.


Antifeedant and insecticidal effects of mandelic acid on the brown planthopper Nilaparvata lugens Stål.
To study the effects of mandelic acid (MA) on the brown planthopper (BPH), Nilaparvata lugens, the survival rate and behaviour of BPH fed on an artificial diet with different dosages of MA was observed. The survival rate of BPH decreased with the increase of the MA concentration and feeding time. In contrast to the control, the survival rate of BPH 72 h after feeding decreased significantly. Electrical penetration graph (EPG) data indicated that MA absorbed by the rice plant from Kimura B solution significantly affected the feeding behaviour of BPH. At the concentrations of 0.1, 0.5, and 1.0 mg/ml, duration of the phloem ingestion of BPH decreased from 115.34 min (control) to 30.41, 7.63, and 0.36 min, respectively. Periods of xylem ingestion of MA-treated BPH were significantly shorter than those of the control (50.44 min). Moreover, BPH spent more time walking around or being at rest on MA-treated rice plants, as well as in stylet activities. The GST (glutathione S-transferase) activity of BPH increased with the increasing MA concentration, while the GPX (glutathione peroxidases) activity did not change significantly. The results indicate that MA has an antifeedant and insecticidal effect on BPH.


Role for actin in the polarized release of rotavirus.
Rotaviruses are characterized by polarized release from the apical side of infected enterocytes, and the rotavirus VP4 spike protein specifically binds to the actin network at the apical pole of differentiated enterocytic cells. To determine the functional consequences of this VP4-actin interaction, fluorescence recovery after photobleaching experiments were carried out to measure the diffusional mobility of VP4 associated with the microfilaments. Results show that VP4 binds to barbed ends of microfilaments by using actin treadmilling. Actin treadmilling inhibition results in the loss of rotavirus apical preferential release, suggesting a major role for actin in polarized rotavirus release.


Identification of a varicella-zoster virus origin of DNA replication and its activation by herpes simplex virus type 1 gene products.
We have identified and characterized an origin of DNA replication in the genome of the human herpesvirus, varicella-zoster virus (VZV). This origin of replication (VZV ORIS) is located within the major inverted repeats in a position equivalent to that occupied by one of the herpes simplex virus type 1 (HSV-1) replication origins. Products encoded by both VZV and HSV-1 activate cloned copies of VZV ORIs, generating high molecular weight molecules consisting of tandem duplications of the input plasmid. The VZV ORIS region contains a tract of alternating A and T residues located at the centre of symmetry of an almost perfect palindrome of 45 bp, and the use of plasmid deletion mutants has demonstrated that this tract is an important functional element of the origin. Two sequences common to the VZV ORIS region and the regions specifying the two HSV-1 origins (ORIS, located within the TRS/IRS regions, and ORIL, located within the UL region) were identified and these may represent important recognition sites. One is an 11 bp sequence (CGTTCGCACTT), and the other is represented by the tract of alternating A and T residues. VZV does not appear to contain an origin of replication in a position equivalent to that of HSV-1 ORIL.


Silicon-based surface plasmon resonance sensing with two surface plasmon polariton modes.
Surface plasmon resonance (SPR) sensing on a silicon-based platform is considered. We have studied properties of SPR in a combined silicon-dielectric layer-gold film-sample medium structure and established conditions of the simultaneous excitation of two plasmon polariton modes that provide narrow and well-separated minima of the reflected intensity. It has been shown that the external mode over the gold-sample medium interface demonstrates a highly sensitive response to a change in the refractive index of the sample medium, whereas the internal mode over the dielectric-gold interface is almost insensitive to medium parameters. We propose that the internal mode can be used as an effective reference zero point for miniature and portable SPR-based systems designed for field and multichannel sensing.


The acetabular rim syndrome. A clinical presentation of dysplasia of the hip.
The acetabular rim syndrome is a pathological entity which we illustrate by reference to 29 cases. The syndrome is a precursor of osteoarthritis of the hip secondary to acetabular dysplasia. The symptoms are pain and impaired function. All our cases were treated by operation which consisted in most instances of re-orientation of the acetabulum by peri-acetabular osteotomy and arthrotomy of the hip. In all cases, the limbus was found to be detached from the bony rim of the acetabulum. In several instances there was a separated bone fragment, or 'os acetabuli' as well. In acetabular dysplasia, the acetabular rim is subject to abnormal stress which may cause the limbus to rupture, and a fragment of bone to separate from the adjacent bone margin. Dysplastic acetabuli may be classified into two radiological types. In type I there is an incongruent shallow acetabulum. In type II the acetabulum is congruent but the coverage of the femoral head is deficient.


Targeting the PDGF signaling pathway in the treatment of non-malignant diseases.
Platelet-derived growth factor (PDGF) is a family of mesenchymal mitogens with important functions during the embryonal development and in the control of tissue homeostasis in the adult. The PDGF isoforms exert their effects by binding to α-and β-tyrosine kinase receptors. Overactivity of PDGF signaling has been linked to the development of certain malignant and non-malignant diseases, including atherosclerosis and various fibrotic diseases. Different types of PDGF antagonists have been developed, including inhibitory monoclonal antibodies and DNA aptamers against PDGF isoforms and receptors, and receptor tyrosine kinase inhibitors. Beneficial effects have been recorded using such inhibitors in preclinical models and in patients with certain malignant as well as non-malignant diseases. The present communication summarizes the use of PDGF antagonists in the treatment of non-malignant diseases.


Focal Type 2 Autoimmune Pancreatitis Mimicking Pancreatic Cancer: Treatment with Neoadjuvant Chemotherapy and Surgery.
A 51-year-old man was referred to our hospital for the further examination of main pancreatic duct interruption. Imaging findings showed a 25-mm-diameter mass lesion located in the pancreatic head. Endoscopic ultrasonography (EUS)-guided fine-needle aspiration (FNA) was performed on the mass. Cytology suggested adenocarcinoma, but the histological diagnosis was not confirmed. We made a comprehensive diagnosis of resectable pancreatic cancer. The mass shrank after preoperative adjuvant chemotherapy, and the patient underwent surgery. The final pathological diagnosis was type 2 autoimmune pancreatitis (AIP). Two years after surgery, AIP had not recurred in the remaining pancreas.


The translational oscillation in oocyte and early embryo development.
Translation is critical for development as transcription in the oocyte and early embryo is silenced. To illustrate the translational changes during meiosis and consecutive two mitoses of the oocyte and early embryo, we performed a genome-wide translatome analysis. Acquired data showed significant and uniform activation of key translational initiation and elongation axes specific to M-phases. Although global protein synthesis decreases in M-phases, translation initiation and elongation activity increases in a uniformly fluctuating manner, leading to qualitative changes in translation regulation via the mTOR1/4F/eEF2 axis. Overall, we have uncovered a highly dynamic and oscillatory pattern of translational reprogramming that contributes to the translational regulation of specific mRNAs with different modes of polysomal occupancy/translation that are important for oocyte and embryo developmental competence. Our results provide new insights into the regulation of gene expression during oocyte meiosis as well as the first two embryonic mitoses and show how temporal translation can be optimized. This study is the first step towards a comprehensive analysis of the molecular mechanisms that not only control translation during early development, but also regulate translation-related networks employed in the oocyte-to-embryo transition and embryonic genome activation.


Pulse Laser Deposition Fabricating Gold Nanoclusters on a Glassy Carbon Surface for Nonenzymatic Glucose Sensing.
A One-step technique for depositing gold nanoclusters (GNCs) onto the surface of a glassy carbon (GC) plate was developed by using pulse laser deposition (PLD) with appropriate process parameters. The method is simple and clean without using any templates, surfactants, or stabilizers. The experimental factors (pulse laser number and the pressure of inert gas (Ar)) that affect the morphology and structure of GNCs, and thus affect the electrocatalytic oxidation performance towards glucose were systematically investigated by means of transmission electron microscopy (TEM) and electrochemical methods (cyclic voltammograms (CV) and chronoamperometry methods). The GC electrode modified by GNCs exhibited a rapid response time (about 2 s), a broad linear range (0.1 to 20 mM), and good stability. The sensitivity was estimated to be 31.18 μA cm(-2) mM(-1) (vs. geometric area), which is higher than that of the Au bulk electrode. It has a good resistance to the common interfering species, such as ascorbic acid (AA), uric acid (UA) and 4-acetaminophen (AP). Therefore, this work has demonstrated a simple and effective sensing platform for the nonenzymatic detection of glucose, and can be used as a new material for a novel non-enzymatic glucose sensor.


Topical dual-probe staining using quantum dot-labeled antibodies for identifying tumor biomarkers in fresh specimens.
Rapid, intra-operative identification of tumor tissue in the margins of excised specimens has become an important focus in the pursuit of reducing re-excision rates, especially for breast conserving surgery. Dual-probe difference specimen imaging (DDSI) is an emerging approach that uses the difference in uptake/clearance kinetics between a pair of fluorescently-labeled stains, one targeted to a biomarker-of-interest and the other an untargeted isotype, to reveal receptor-specific images of the specimen. Previous studies using antibodies labeled with either enhanced Raman particles or organic fluorophores have shown promising tumor vs. normal diagnostic performance. Yet, the unique properties of quantum dot-labeled antibody complexes (QDACs), which provide spectrally-distinct fluorescence emission from a common excitation source, make them ideal candidates for this application. Herein, we evaluate the diagnostic performance of QDAC-based DDSI in excised xenografts. Excised fresh specimens of normal tissue and human tumor xenografts with elevated expression of HER2 were stained with a HER2-targeted QDAC and an untargeted QDAC isotype. Stained specimens were imaged on a custom hyperspectral imaging system capable of spectrally separating the quantum dot signatures, and images processed using the DDSI approach. The diagnostic performance of this technique under different incubation temperatures and probe concentrations was evaluated using receiver-operator characteristic analysis. HER2-targeted QDAC-DDSI was able to distinguish HER2(+) tumors from normal tissue with reasonably high diagnostic performance; however, this performance was sensitive to temperature during the staining procedure. Area under the curve values were 0.61 when staining at room temperature but increased to over 0.81 when staining at 37 °C. Diagnostic performance was not affected by increasing stain concentration. This study is the first to report dual-probe difference imaging of specimens using QDACs and hyperspectral imaging. Our results show promising diagnostic performance under certain conditions, and compel further optimization and evaluation of this intra-operative margin assessment technique.


Pharmacokinetic and pharmacodynamic analysis of efavirenz dose reduction using an in vitro-in vivo extrapolation model.
The pharmacokinetics (PK) of efavirenz (EFV) is characterized by marked interpatient variability that correlates with its pharmacodynamics (PD). In vitro-in vivo extrapolation (IVIVE) is a "bottom-up" approach that combines drug data with system information to predict PK and PD. The aim of this study was to simulate EFV PK and PD after dose reductions. At the standard dose, the simulated probability was 80% for viral suppression and 28% for central nervous system (CNS) toxicity. After a dose reduction to 400 mg, the probabilities of viral suppression were reduced to 69, 75, and 82%, and those of CNS toxicity were 21, 24, and 29% for the 516 GG, 516 GT, and 516 TT genotypes, respectively. With reduction of the dose to 200 mg, the probabilities of viral suppression decreased to 54, 62, and 72% and those of CNS toxicity decreased to 13, 18, and 20% for the 516 GG, 516 GT, and 516 TT genotypes, respectively. These findings indicate how dose reductions might be applied in patients with favorable genetic characteristics.


Almotriptan for the treatment of acute migraine: a review of early intervention trials.
Almotriptan is a serotonin (5-hydroxytryptamine)(1B/1D) receptor agonist (triptan) that has shown consistent efficacy in the acute treatment of migraine with excellent tolerability. It is an effective, well-tolerated and cost-effective triptan, as demonstrated by improvement in rigorous, patient-orientated end points, such as 'sustained pain-free without adverse events'. Results from post hoc analyses, observational studies and well-controlled, prospective clinical trials have shown that significant improvements can be achieved if almotriptan 12.5 mg is administered within an hour of migraine onset, particularly when pain is mild, rather than waiting until pain is moderate-to-severe. Benefits were also achieved with early treatment of moderate-to-severe pain. Time-to-treatment was the best predictor of headache duration, whereas initial headache intensity best predicted most other efficacy outcomes. Early administration of almotriptan 12.5 mg not only produced rapid symptomatic relief, it also improved the patient's quality of life and ability to resume normal daily functioning. Furthermore, the efficacy of almotriptan is not significantly affected by allodynia (purported to reduce the efficacy of triptans). Thus, the excellent efficacy and tolerability profile of almotriptan administered early in a migraine attack indicate that it may be a first-line treatment option in this common, underdiagnosed and undertreated disorder.


MMPI inmate profiles: suicide completers, suicide attempters, and non-suicidal controls.
Results from the Minnesota Multiphasic Personality Inventory (MMPI) were compared for three groups of male inmates in federal penitentiaries: 47 suicide completers, 43 suicide attempters and, 123 non-suicidal controls. Analyses show that the groups differed on all 10 clinical scales and on at least 8 after Bonferroni correction. Attempters obtained the highest scores on 4 of 8 scales; they also posted the highest scores on 3 others, but differences were significant only against non-suicidal controls. Completers posted the highest score on only one scale (Mf), but the difference was significant only against non-suicidal controls. All told, completers proved more similar to non-suicidal controls than to attempters. Regarding profiles, completers are more strongly correlated with non-suicidal controls (r=0.95) than with attempters (r=0.86); non-suicidal controls are less strongly correlated with attempters (r=0.88). The fact that attempters seem more pathological than the others can mislead clinicians screening for suicide risk.


Hippocampal volume, PTSD, and alcoholism in combat veterans.
Studies imposing rigorous control over lifetime alcohol intake have usually not found smaller hippocampal volumes in persons with posttraumatic stress disorder. Because the majority of negative studies have used adolescent samples, it has been suggested that chronicity is a necessary condition for such findings. To test the hypothesis that a smaller hippocampus in PTSD is unrelated to comorbid alcoholism or to chronicity, this study estimated hippocampal volume in a relatively large group (N=99) of combat veterans in which PTSD, lifetime alcohol abuse/dependence, and Vietnam versus Gulf War service were crossed. In subjects with histories of alcoholism, unadjusted hippocampal volume was 9% smaller in persons with PTSD than in those without PTSD. In nonalcoholic subjects, the PTSD-related difference in hippocampal volume was 3%. The failure to observe a strong association between PTSD and hippocampal volume in nonalcoholic subjects was not ascribable to younger age, reduced PTSD chronicity, or lower PTSD symptom severity. The possibility that smaller hippocampal volume is limited to groups in which PTSD is compounded by comorbid alcoholism is not necessarily incompatible with results suggesting a smaller hippocampus is predispositional to PTSD. Further examination of the role of alcoholism and other comorbid conditions in studies of brain structure and function in PTSD appears warranted.


[Being a woman with HPV: a cultural approach].
This study was carried out in order to investigate the level of women's knowledge about HPV, to identify beliefs, myths and taboos about HPV, as well as to analyze the influence of these cultural elements on women's behavior. We performed a study based on the transcultural care theory. The research was developed from the testimonies of 15 women who were undergoing treatment for HPV. Through the investigation, it was possible to perceive that HPV is a disease that is still unknown and surrounded with mystery, despite all the sources of information about Sexually Trasmitted Diseases (STD). This ignorance, which interacts with cultural factors, favors the development of wrong concepts, such as beliefs and myths.


Patient-Specific Vascularized Tumor Model: Blocking TAM Recruitment with Multispecific Antibodies Targeting CCR2 and CSF-1R.
Tumor-associated inflammation drives cancer progression and therapy resistance, with the infiltration of monocyte-derived tumor-associated macrophages (TAMs) associated with poor prognosis in diverse cancers. Targeting TAMs holds potential against solid tumors, but effective immunotherapies require testing on immunocompetent human models prior to clinical trials. Here, we develop an in vitro model of microvascular networks that incorporates tumor spheroids or patient tissues. By perfusing the vasculature with human monocytes, we investigate monocyte trafficking into the tumor and evaluate immunotherapies targeting the human tumor microenvironment. Our findings demonstrate that macrophages in vascularized breast and lung tumor models can enhance monocyte recruitment via TAM-produced CCL7 and CCL2, mediated by CSF-1R. Additionally, we assess a novel multispecific antibody targeting CCR2, CSF-1R, and neutralizing TGF-β, referred to as CSF1R/CCR2/TGF-β Ab, on monocytes and macrophages using our 3D models. This antibody repolarizes TAMs towards an anti-tumoral M1-like phenotype, reduces monocyte chemoattractant protein secretion, and effectively blocks monocyte migration. Finally, we show that the CSF1R/CCR2/TGF-β Ab inhibits monocyte recruitment in patient-specific vascularized tumor models. Overall, this vascularized tumor model offers valuable insights into monocyte recruitment and enables functional testing of innovative therapeutic antibodies targeting TAMs in the tumor microenvironment (TME).


Cellulose nanofibrils improve dispersibility and stability of silver nanoparticles and induce production of bacterial extracellular polysaccharides.
Many polymer-stabilized silver nanoparticles (AgNPs) with enhanced antibacterial properties have been synthesized, but very little is known about the fate of these materials and their interactions with microbes in physiological solutions. In this study, we evaluated the role of cellulose nanofibrils (CNFs) in stabilizing AgNPs (CNF-AgNPs) in a bacterial growth medium, determined the antibacterial properties of CNF-AgNPs and assessed the CNF and CNF-AgNP interactions with bacteria. The attachment of AgNPs to CNFs significantly improved the stability and dispersibility of AgNPs in the bacterial growth medium compared to colloidal AgNPs. The CNF-AgNPs exerted a concentration-dependent growth-inhibitory effect on E. coli. An extracellular polysaccharide (EPS)-like structure was formed around the E. coli bacterium when incubated with a sub-lethal CNF-AgNP concentration, which led to the clustering of neighboring bacteria. No EPS-like structure was observed around the bacterium incubated with high concentration of CNF-AgNPs. Overall, this study demonstrated that CNFs significantly improved the stability and dispersibility of AgNPs in physiological medium, validated the antimicrobial potential of CNF-AgNPs, and provided an insight into the physio-chemical interactions between bacteria and CNF-AgNPs in a physiological growth medium.


Accounting for spatial sampling patterns in Bayesian phylogeography.
Statistical phylogeography provides useful tools to characterize and quantify the spread of organisms during the course of evolution. Analyzing georeferenced genetic data often relies on the assumption that samples are preferentially collected in densely populated areas of the habitat. Deviation from this assumption negatively impacts the inference of the spatial and demographic dynamics. This issue is pervasive in phylogeography. It affects analyses that approximate the habitat as a set of discrete demes as well as those that treat it as a continuum. The present study introduces a Bayesian modeling approach that explicitly accommodates for spatial sampling strategies. An original inference technique, based on recent advances in statistical computing, is then described that is most suited to modeling data where sequences are preferentially collected at certain locations, independently of the outcome of the evolutionary process. The analysis of georeferenced genetic sequences from the West Nile virus in North America along with simulated data shows how assumptions about spatial sampling may impact our understanding of the forces shaping biodiversity across time and space.


[Physical therapy in the complex treatment of thyroiditis].
Pathogenetic approaches (clinico-anatomic, structural features of thyroid epithelium, echographic signs, functional activity of the thyroid) served the basis for division of inflammatory diseases into 4 groups. A wide spectrum of physiotherapeutic procedures are suggested for treatment of thyroiditis. There may be different combination of physiotherapy with medication. The response was assessed by ultrasonography and dopplerography evidence. Physiotherapy potentiates the action of drugs and can be also used as monotherapy in dysfunctional thyroid pathology.


Large-Conductance Calcium-Activated Potassium Channels and Voltage-Dependent Sodium Channels in Human Cementoblasts.
Cementum, which is excreted by cementoblasts, provides an attachment site for collagen fibers that connect to the alveolar bone and fix the teeth into the alveolar sockets. Transmembrane ionic signaling, associated with ionic transporters, regulate various physiological processes in a wide variety of cells. However, the properties of the signals generated by plasma membrane ionic channels in cementoblasts have not yet been described in detail. We investigated the biophysical and pharmacological properties of ion channels expressed in human cementoblast (HCEM) cell lines by measuring ionic currents using conventional whole-cell patch-clamp recording. The application of depolarizing voltage steps in 10 mV increments from a holding potential (Vh) of -70 mV evoked outwardly rectifying currents at positive potentials. When intracellular K+ was substituted with an equimolar concentration of Cs+, the outward currents almost disappeared. Using tail current analysis, the contributions of both K+ and background Na+ permeabilities were estimated for the outward currents. Extracellular application of tetraethylammonium chloride (TEA) and iberiotoxin (IbTX) reduced the densities of the outward currents significantly and reversibly, whereas apamin and TRAM-34 had no effect. When the Vh was changed to -100 mV, we observed voltage-dependent inward currents in 30% of the recorded cells. These results suggest that HCEM express TEA- and IbTX-sensitive large-conductance Ca2+-activated K+ channels and voltage-dependent Na+ channels.


Evaluation of the efficacy of cystinosin supplementation through CTNS mRNA delivery in experimental models for cystinosis.
Messenger RNA (mRNA) therapies are emerging in different disease areas, but have not yet reached the kidney field. Our aim was to study the feasibility to treat the genetic defect in cystinosis using synthetic mRNA in cell models and ctns-/- zebrafish embryos. Cystinosis is a prototype lysosomal storage disorder caused by mutations in the CTNS gene, encoding the lysosomal cystine-H+ symporter cystinosin, and leading to cystine accumulation in all cells of the body. The kidneys are the first and the most severely affected organs, presenting glomerular and proximal tubular dysfunction, progressing to end-stage kidney failure. The current therapeutic standard cysteamine, reduces cystine levels, but has many side effects and does not restore kidney function. Here, we show that synthetic mRNA can restore lysosomal cystinosin expression following lipofection into CTNS-/- kidney cells and injection into ctns-/- zebrafish. A single CTNS mRNA administration decreases cellular cystine accumulation for up to 14 days in vitro. In the ctns-/- zebrafish, CTNS mRNA therapy improves proximal tubular reabsorption, reduces proteinuria, and restores brush border expression of the multi-ligand receptor megalin. Therefore, this proof-of-principle study takes the first steps in establishing an mRNA-based therapy to restore cystinosin expression, resulting in cystine reduction in vitro and in the ctns-/- larvae, and restoration of the zebrafish pronephros function.


The distinct functional properties of the nucleotide-binding domain of ATP7B, the human copper-transporting ATPase: analysis of the Wilson disease mutations E1064A, H1069Q, R1151H, and C1104F.
Copper transport by the P(1)-ATPase ATP7B, or Wilson disease protein (WNDP),1 is essential for human metabolism. Perturbation of WNDP function causes intracellular copper accumulation and severe pathology, known as Wilson disease (WD). Several WD mutations are clustered within the WNDP nucleotide-binding domain (N-domain), where they are predicted to disrupt ATP binding. The mechanism by which the N-domain coordinates ATP is presently unknown, because residues important for nucleotide binding in the better characterized P(2)-ATPases are not conserved within the P(1)-ATPase subfamily. To gain insight into nucleotide binding under normal and disease conditions, we generated the recombinant WNDP N-domain and several WD mutants. Using isothermal titration calorimetry, we demonstrate that the N-domain binds ATP in a Mg(2+)-independent manner with a relatively high affinity of 75 microm, compared with millimolar affinities observed for the P(2)-ATPase N-domains. The WNDP N-domain shows minimal discrimination between ATP, ADP, and AMP, yet discriminates well between ATP and GTP. Similar results were obtained for the N-domain of ATP7A, another P(1)-ATPase. Mutations of the invariant WNDP residues E1064A and H1069Q drastically reduce nucleotide affinities, pointing to the likely role of these residues in nucleotide coordination. In contrast, the R1151H mutant exhibits only a 1.3-fold reduction in affinity for ATP. The C1104F mutation significantly alters protein folding, whereas C1104A does not affect the structure or function of the N-domain. Together, the results directly demonstrate the phenotypic diversity of WD mutations within the N-domain and indicate that the nucleotide-binding properties of the P(1)-ATPases are distinct from those of the P(2)-ATPases.


The role of the gene 71 product in the life cycle of equine herpesvirus 1.
Equine herpesvirus type 1 (EHV-1) gene 71 encodes a heavily O-glycosylated 192 kDa protein with no identified herpesvirus homologue. Isolation of a deletion mutant in gene 71 (ED71) demonstrated that its protein product is not essential in vitro. To investigate the role of the gene 71 protein in the virus life cycle, ED71 has been characterized in vitro in terms of cellular adsorption, penetration, egress and transmission compared to wild-type and revertant virus. ED71 virions adsorbed to cells less efficiently than wild-type and revertant virus with a consequential effect on virus penetration; virus egress was significantly impaired and the timing of release was also delayed. The percentage of both full and empty capsids accumulating in the nuclei of ED71-infected cells was significantly higher than in wild-type virus-infected cells but the most notable differences were the low number of particles and the low ratio of enveloped to unenveloped capsids in the cytoplasm. The primary mode of transmission of the mutant virus is by direct cell-to-cell spread and the fact that a neutralizing antiserum did not reduce ED71 plaque size, supported the conclusion that deletion of gene 71 impairs the ability of virus to spread via release and readsorption to uninfected cells. Thus, deletion of EHV-1 gene 71 results in a defect in virus maturation and capsid envelopment. Progeny virus is consequently impaired in adsorption/penetration presumably due to the particles lacking the glycoprotein spikes predicted to be encoded by this gene and hence spreads by direct cell-to-cell contact.


Plasma cytomegalovirus (CMV) DNA load predicts CMV disease and survival in AIDS patients.
In this study, baseline plasma from 619 persons with acquired immunodeficiency syndrome (AIDS) (median CD4+ lymphocyte count -21/microl) who participated in a trial to determine the efficacy of oral ganciclovir for cytomegalovirus (CMV) disease prevention were evaluated for CMV DNA load by qualitative and quantitative polymerase chain reaction (PCR), and correlated with the development of CMV disease and survival. For participants without detectable plasma CMV DNA, the 12-mo Kaplan-Meier CMV disease event rate was 14% and 1% for the placebo and ganciclovir groups, respectively (P < 0.001). For PCR positive participants, CMV disease developed in 43% of placebo and 26% ganciclovir recipients (P < 0.017). Among placebo recipients, CMV PCR positivity was associated with a 3.4-fold increased risk of developing CMV disease (P < 0.001) whereas CD4+ lymphocyte count was not a useful predictor (P = 0.47). A positive plasma CMV DNA PCR was also associated with a 2.5-fold increased risk of death. Each log10 increase in baseline CMV DNA load was associated with a 3.1-fold increase in CMV disease (P < 0.001) and a 2.2-fold increase in mortality (P < 0.001). These data indicate that the risk of developing CMV disease and death in persons with advanced AIDS is directly related to the quantity of CMV DNA in plasma, and is a better predictor than CD4+ lymphocyte count in this population.


Prevalence of Mimics and Severe Comorbidity in Patients with Clinically Suspected Transient Global Amnesia.
Transient global amnesia (TGA) is a syndrome featuring acute anterograde amnesia as the most striking clinical symptom. Its etiology is still a matter of debate. Most neurological guidelines allow the diagnosis on the basis of clinical criteria only; a more extensive evaluation is recommended only for patients with "red flags" like severe headache, nausea or vomiting, or metabolic abnormalities. The aim of our study was to assess the frequency of a severe underlying disease or alternative diagnoses (mimics) in patients fulfilling the clinical criteria. We evaluated the medical records and the imaging data of an unselected consecutive cohort of patients with suspected TGA over a 7-year period. All patients were hospitalized and received a neurological workup including brain imaging, color-coded duplex sonography of the brain supplying arteries, electroencephalography, and laboratory studies of blood and (in selected cases) cerebrospinal fluid. 163 patients with 166 episodes of suspected TGA were hospitalized (3 patients twice). After the workup, the diagnosis of TGA was confirmed in 148/166 (89.2%) episodes ("simple TGA"). Eighteen patients (10.8%) either had an alternative diagnosis or a severe comorbidity that was assumed to have had an impact on the occurrence of the amnestic episode ("complicated TGA/mimic"). The most important differential diagnosis was stroke (11 patients, 6.6% of all TGA suspects and 61.1% of the complicated TGA/mimic group). Other mimics were transient epileptic amnesia (2 patients) and steroid-induced delirium (1 patient). Important comorbidities that had not been obvious at the time of presentation were severe sleep apnea (2 patients), triptan overuse (1 patient), and an involuntary amlodipine intoxication during TGA. As approximately every tenth patient with suspected TGA either had an alternative diagnosis or a severe comorbidity, which had not been obvious at the time of admission, we consider in-patient treatment of all suspected TGA cases as appropriate, preferably in the setting of a stroke unit, as ischemic stroke was the by far most important diagnosis mimicking TGA.


Liquid phase immunoassays utilizing magnetic markers and SQUID magnetometer.
Immunoassays are one main detection system used in the field of clinical chemistry. Recent developments of a new detection method utilizing a magnetic marker and magnetic sensor have enabled rapid and sensitive immunoassay without the need for bound/free (BF) separation. Newly-synthesized conjugated avidin was used as the magnetic marker for quantitative analysis of human interleukin-8 (hIL-8) and immunoglobulin E (hIgE) in several media. A superconducting quantum interference device sensor detected the magnetic fields from markers fixed to antigens by the sandwich method. Magnetic signals from unbound markers were nearly zero due to Brownian rotation. Our magnetic immunoassay could detect four attomoles of model proteins (hIL-8, hIgE) in phosphate buffer without BF separation. Using our standard curve, the range of protein detected ranged from 40 femtomoles to 4 attomoles, and we observed a strong association between protein amounts and magnetic signals from the bound markers. The homogeneous immunoassay could also quantify three hundred cells from the fungus Candida albicans in phosphate buffer. The present study demonstrates the ability of magnetic markers for measuring biological targets without BF separation. This detection system has great potential for use as the next generation's analytical system.


Controlling gene expression with 2-5A antisense.
Recent work has demonstrated that the activity of a ubiquitous cellular enzyme, ribonuclease L (RNase L), can be harnessed to cleave targeted RNA species. Activation of RNase L is dependent on the presence of 2',5'-linked oligoadenylates (2-5A), usually produced by cells infected with viruses. By conjugating synthetic 2-5A to specific antisense compounds, it is now possible to selectively degrade RNAs in an RNase L-dependent manner, thereby providing an alternative to RNase H-dependent approaches. In this summary, we provide an updated description of the synthesis procedure for constructing these chimeric 2-5A antisense molecules. Examples of successful applications of the 2-5A antisense strategy are described, along with some of the procedures involved in those studies. Several methods are also provided for optimizing compound uptake and analyzing their effects on cells. Finally, we discuss the current body of evidence that supports the contention that RNase L is indeed the primary mediator of 2-5A antisense effects and the possible implications that this has on the future of this therapeutic approach.


Limitations of the fasting proinsulin to insulin ratio as a measure of β-cell health in people with and without impaired glucose tolerance.
The fasting proinsulin to insulin ratio is elevated in people with type 2 diabetes and has been suggested as a marker of β-cell health. However, its utility in discriminating between individuals with varying degrees of β-cell dysfunction is unclear. Proinsulin has a very different half-life to insulin and unlike insulin does not undergo hepatic extraction prior to reaching the systemic circulation. Given these limitations, we sought to examine the relationship between fasting and postprandial concentrations of β-cell polypeptides (proinsulin, insulin and C-peptide) in people with normal and impaired glucose tolerance in differing metabolic environments. Subjects were studied on two occasions in random order while undergoing an oral challenge. During one study day, free fatty acids were elevated (to induce insulin resistance) by infusion of Intralipid with heparin. Proinsulin to insulin and proinsulin to C-peptide ratios were calculated for the 0-, 30-, 60- and 240-minute time points. Insulin action (Si) and β-cell responsivity (Φ) indices were calculated using the oral minimal model. The fasting proinsulin to c-peptide or fasting proinsulin to insulin ratios did not differ between groups and did not predict subsequent β-cell responsivity to glucose during the glycerol or Intralipid study days in either group. Among nondiabetic individuals, the fasting proinsulin to insulin ratio is not a useful marker of β-cell function.


Malignant cell-derived extracellular vesicles express different chromogranin epitopes compared to prostasomes.
Prostasomes are nanosized extracellular vesicles exocytosed by prostate epithelial cells. They have been assigned many roles propitious to sperm in favor of fertilization. Prostatic cancer cells can also produce and secrete extracellular vesicles. We assessed using ELISA, the surface expression of chromogranin proproteins on prostasomes and malignant extracellular vesicles of four different prostate cancer cell-lines, two hormone sensitive and two hormone refractory. We used a panel of chromogranin A and chromogranin B antibodies against peptides in-between hypothetical cleavage sites along the proproteins. A diverging pattern of chromogranin peptides was apparent when comparing prostasomes and malignant extracellular vesicles indicating a phenotypical change. We also compared western blot patterns (prostasomes and malignant extracellular vesicles) for selected antibodies that displayed high absorbances in the ELISA. Western blot analyses revealed various cleavage patterns of those proproteins that were analyzed in prostasomes and extracellular vesicles. Chromogranins are constituents of not only prostasomes but also of malignant prostate cell-derived extracellular vesicles with different amino acid sequences exposed at the membrane surface giving rise to a mosaic pattern. These findings may be of relevance for designing new assays for detection or even possible treatment of prostate cancers.


Types of interventions made by clinical pharmacists in the IMPROVE study. Impact of Managed Pharmaceutical Care on Resource Utilization and Outcomes in Veterans Affairs Medical Centers.
The purpose of this study was to describe and evaluate the activities and interventions provided by ambulatory care clinical pharmacists during the IMPROVE (Impact of Managed Pharmaceutical Care on Resource Utilization and Outcomes in Veterans Affairs Medical Centers) study. A total of 523 patients were randomized into the intervention arm at nine Veterans Affairs medical centers if they were considered to be at high risk for drug-related problems. Patients randomized to the control group had no interventions and they are not reported. Using a standard form, pharmacists were asked to document the length of visit, method of contact, medical conditions addressed, and drug-related problems addressed and resolved during each contact. Seventy-eight ambulatory care clinical pharmacists documented 1855 contacts over 12 months, an average of 3.54 +/- 2.31/patient. The length of visits was 15 minutes or more for 73% of contacts. In-person contacts accounted for 1421 visits (76.6%), with the remainder being telephone contacts. During each contact the average number of drug-related problems addressed and resolved were 1.64 +/- 1.16 and 1.14 +/- 0.98, respectively. More drug-related problems were addressed and resolved when visits were 15 minutes or longer (p=0.001) and when the contact was in person (p=0.001). These data may provide information to clinical pharmacists developing pharmacy-managed clinics for patients at high risk for drug-related problems. The information may be a benchmark for types of interventions that can be made, as well as the time commitments required to make them.


High-level expression and characterization of Galactomyces geotrichum (BT107) lipase I in Pichia pastoris.
The mature lipI gene, encoding the lipase I from Galactomyces geotrichum BT107, was obtained by PCR from genomic DNA, sequenced and cloned into a Pichia pastoris expression vector. Clones containing multiple copies of lipI integrated in their genome were analyzed to achieve high-level expression of the recombinant lipase I. One strain with four or more copies of the expression cassette was able to produce more than 200mg/L of extracellular heterologous protein. The lipase I was partially purified using anion exchange chromatography and its activity on monounsaturated (triolein) and polyunsaturated (triEPA) triglycerides was analyzed by a novel HPLC-MS assay.


Magnetic resonance imaging markers for early diagnosis of Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by selective and progressive degeneration, as well as loss of dopaminergic neurons in the substantia nigra. In PD, approximately 60-70% of nigrostriatal neurons are degenerated and 80% of content of the striatal dopamine is reduced before the diagnosis can be established according to widely accepted clinical diagnostic criteria. This condition describes a stage of disease called "prodromal", where non-motor symptoms, such as olfactory dysfunction, constipation, rapid eye movement behaviour disorder, depression, precede motor sign of PD. Detection of prodromal phase of PD is becoming an important goal for determining the prognosis and choosing a suitable treatment strategy. In this review, we present some non-invasive instrumental approaches that could be useful to identify patients in the prodromal phase of PD or in an early clinical phase, when the first motor symptoms begin to be apparent. Conventional magnetic resonance imaging (MRI) and advanced MRI techniques, such as magnetic resonance spectroscopy imaging, diffusion-weighted and diffusion tensor imaging and functional MRI, are useful to differentiate early PD with initial motor symptoms from atypical parkinsonian disorders, thus, making easier early diagnosis. Functional MRI and diffusion tensor imaging techniques can show abnormalities in the olfactory system in prodromal PD.


High-Altitude Pulmonary Edema Combined with Spontaneous Pneumomediastinum: A Case Report.
High-altitude pulmonary edema (HAPE) is a serious life-threatening disease that occurs after rapid ascent to high altitude; its main early-stage presentations include fatigue, headache, low-grade fever, dyspnea, and cough. X-ray and computed tomography (CT) images show pulmonary shadows and patches, which may be localized (initial right lung field predomination) or generalized to the bilateral lung base. In this report, we present a case of a 25-year-old man diagnosed with HAPE combined with spontaneous pneumomediastinum. After a quick descent and effective medical treatment, this patient made a full recovery. The case may provide helpful information for the prevention and treatment of this disease since an increased number of people, especially young men, currently travel and work at high altitudes. After accurate clinical diagnosis with the help of CT or X-ray, immediate descent and appropriate oxygen supplementation are the most effective treatments for HAPE at high altitude.


Parametric Loop Division for 3D Localization in Wireless Sensor Networks.
Localization in Wireless Sensor Networks (WSNs) has been an active topic for more than two decades. A variety of algorithms were proposed to improve the localization accuracy. However, they are either limited to two-dimensional (2D) space, or require specific sensor deployment for proper operations. In this paper, we proposed a three-dimensional (3D) localization scheme for WSNs based on the well-known parametric Loop division (PLD) algorithm. The proposed scheme localizes a sensor node in a region bounded by a network of anchor nodes. By iteratively shrinking that region towards its center point, the proposed scheme provides better localization accuracy as compared to existing schemes. Furthermore, it is cost-effective and independent of environmental irregularity. We provide an analytical framework for the proposed scheme and find its lower bound accuracy. Simulation results shows that the proposed algorithm provides an average localization accuracy of 0.89 m with a standard deviation of 1.2 m.


Primary weight loss failure after Roux-en-Y gastric bypass is characterized by impaired gut-hormone mediated regulation of food intake.
After Roux-en-Y gastric bypass (RYGB) a subset of patients never obtain excess BMI loss (EBMIL) > 50% and are categorized as having primary weight loss (WL) failure. We hypothesized that postprandial concentrations of glucagon-like peptide 1 (GLP-1) and peptide YY (PYY) would be lower in patients with primary WL failure compared with patients with successfully maintained WL. Furthermore, that inhibition of gut hormone secretions would increase ad libitum food intake less in patients with primary WL failure. Twenty women with primary WL failure (LowEBMIL < 50%) were individually matched to twenty women with successful WL (HighEBMIL > 60%) on age, preoperative BMI and time from RYGB. On separate days performed in a random order, patient-blinded subcutaneous injections of octreotide or saline (placebo) were followed by a fixed breakfast and an ad libitum lunch with blood sampling for appetite regulating hormones and Visual-Analogue-Scale (VAS)-scoring of hunger/satiety. Furthermore, participants underwent gene variant analysis for GLP-1, PYY and their receptors, indirect calorimetry, dual-energy X-ray absorptiometry (DXA)-scans, 4-days at-home food registration and 14-days step counting. On placebo days, postprandial GLP-1, PYY and cholecystokinin (CCK) concentrations were similar between groups after breakfast. Fasting ghrelin was lower in LowEBMIL, but the postprandial suppression was similar. LowEBMIL had lower satiety VAS-scores and less suppression of hunger VAS-scores. Gene variants did not differ between groups. Octreotide diminished GLP-1, PYY, CCK and ghrelin concentrations in both groups. Octreotide did not affect ad libitum food intake in LowEBMIL (-1% [-13, 12], mean [95%CI]), while food intake increased in HighEBMIL (+23% [2,44]). Primary WL failure after RYGB was not characterized by impaired secretions of appetite regulating gut hormones. Interestingly, inhibition of gut hormone secretions with octreotide only increased food intake in patients with successful WL post-RYGB. Thus, an impaired central anorectic response to gut hormones may contribute to primary WL failure after RYGB.