Temperature dependent charge transport across tunnel junctions of single-molecules and self-assembled monolayers: a comparative study.
In this work we present a comparative study of the temperature behavior of charge current in both single-molecule transistors and self-assembled monolayer-based tunnel junctions with symmetrical molecules of alkanethiolates functionalized with a ferrocene (Fc) unit. The Fc unit is separated from the electrodes with two equal alkyl chains of enough length to ensure weak coupling of the Fc unit with the electrodes. These junctions do not rectify charge current and display exponential dependence with temperature with moderate slopes, which can be directly associated to the thermal broadening of the electronic occupation Fermi distribution in the electrodes. The capability to electrically gate the molecular frontier orbital of the Fc (here the highest occupied molecular orbital, HOMO) in the single-molecule transistor, not possible in the two-terminal SAM-based junctions, allows for a detailed comparative between the two classes of junctions. Our findings demonstrate that, although most transport characteristics are equivalent, collective effects arising from interactions between molecules in the self-assembled monolayer greatly affect the energetics of SAM-based junctions, resulting in a bias-independent tunnel current, contrary to the case of the single-molecule junction and as expected from the thermal broadening of the electronic occupation around the Fermi energy in the electrodes.


NTP Toxicology and Carcinogenesis Studies of Isoprene (CAS No. 78-79-5) in F344/N Rats (Inhalation Studies).
Isoprene, the monomeric unit of natural rubber and naturally occurring terpenes and steroids, is primarily obtained as a by-product of naphtha cracking for ethylene production. It is emitted from plants and trees, has been detected in tobacco smoke and automobile exhaust, and was identified as a major endogenous hydrocarbon in human breath. Isoprene was selected for toxicologic evaluation because of its structural similarity to 1,3-butadiene, a potent, multi-organ, rodent carcinogen, and the potential for human exposure due to its large annual production volume. A previous 26-week inhalation study followed by a 26-week recovery period provided clear evidence of carcinogenic activity of isoprene in male B6C3F1 mice. A similar study in male F344/N rats was inconclusive. Male and female F344/N rats were exposed to isoprene (99% pure) by whole body inhalation for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, mouse bone marrow and peripheral blood cells, and rat lung fibroblasts. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 220, 700, or 7,000 ppm isoprene by inhalation, 6 hours per day, 5 days per week, for 105 weeks. Survival and Body Weights: Survival rates and mean body weights of exposed male and female rats were similar to those of the chamber controls throughout the study. Urinary Vinyl Lactic Acid - Biomarker of Exposure: At 3, 6, 12, and 18 months, the concentrations of vinyl lactic acid normalized to creatinine in the urine increased with increasing exposure concentration in all exposed groups of male and female rats; however, these increases were not proportional to isoprene exposure concentrations, indicating nonlinear metabolism over this range of exposure concentrations. Pathology Findings: Exposure-related increases in the incidences of mam mary gland fibroadenoma were observed in male rats in all groups. The incidences of fibroadenoma in 7,000 ppm males and in all groups of exposed females were significantly greater than those in the chamber control groups. The incidences of fibroadenoma in all exposed groups of males and females and of multiple fibroadenoma in 7,000 ppm males and in all groups of exposed females exceeded the historical control ranges. In addition, the finding of mammary gland carcinoma in exposed male rats was noteworthy because this neoplasm rarely occurs in control male rats. The incidences of renal tubule adenoma in 700 and 7,000 ppm males and of renal tubule hyperplasia in 7,000 ppm males were significantly greater than those in the chamber controls. The severity of kidney nephropathy was slightly increased in 7,000 ppm males when compared to chamber controls. An exposure-related increase in the incidences of interstitial cell adenoma of the testis was observed in male rats. The incidences of bilateral interstitial cell adenoma and of unilateral and bilateral interstitial cell adenoma (combined) of the testis in 700 and 7,000 ppm males were significantly greater than those in the chamber controls. The incidences of interstitial cell adenoma in 700 and 7,000 ppm males exceeded the historical control range. Several rare neoplasms including benign astrocytoma, malignant glioma, malignant medulloblastoma be nign meningeal granular cell tumor, and meningeal sarcoma were observed in the brain of exposed female rats. These neoplasms have seldom or never occurred in historical chamber controls. The incidences of splenic fibrosis in 700 and 7,000 ppm males were significantly greater than that in the chamber control group. GENETIC TOXICOLOGY: Isoprene was not mutagenic in S. typhimurium and did not induce sister chromatid exchanges or chromosomal aberrations in cultured Chinese hamster ovary cells with or without exogenous metabolic activation; however, in mice, isoprene induced increases in the frequency of sister chromatid exchanges in bone marrow cells and in the frequency of micronucleated erythrocytes in peripheral blood. The cell cycle duration of proliferating bone marrow cells of mice f mice exposed to 7,000 ppm isoprene was significantly lengthened. No increases in the frequency of chromosomal aberrations were observed in bone marrow cells of male mice after 12 days of exposure to isoprene, and lung fibroblasts of male and female rats exposed to isoprene for 4 weeks showed no increase in the frequency of micronuclei. CONCLUSIONS: Under the conditions of this 2-year inhalation study, there was clear evidence of carcinogenic activity of isoprene in male F344/N rats based on increased incidences of mammary gland fibroadenoma and carcinoma, renal tubule adenoma, and testicular interstitial cell adenoma. There was some evidence of carcinogenic activity of isoprene in female F344/N rats based on increased incidences and multiplicity of mammary gland fibroadenoma. A low incidence of rare brain neoplasms in exposed female rats may have been due to exposure to isoprene. Exposure to isoprene by inhalation for 2 years resulted in increased incidences of renal tubule hyperplasia and splenic fibrosis in male rats. Synonyms: Isopentadiene; b-methylbivinyl; 2-methyl-1,3-butadiene


Adult acute megakaryocytic leukemia: an analysis of 37 patients treated at M.D. Anderson Cancer Center.
To characterize acute megakaryocytic leukemia (FAB M7 AML), we identified 37 patients with M7 AML treated at M.D. Anderson Cancer Center between 1987 and 2003 and compared them with 1800 patients with non-M7, non-M3 AML treated during the same period. The median age of the M7 AML group was 56 years (range, 21-78 years); 22 patients (59%) had an antecedent hematologic disorder or myelodysplastic syndrome or both, and 7 patients (19%) had previously received chemotherapy for other malignancies. Extensive bone marrow fibrosis was found in 23 patients (62%). Poor cytogenetic characteristics were observed in 49% of patients with M7 AML versus 33% of others (P < .001). Complete remission rates were 43% with M7 AML and 57% with non-M7 AML (P = .089). Median overall survival (OS) was 23 and 38 weeks, respectively (P = .006). Median disease-free survivals were 23 versus 52 weeks, respectively (P < .001). By multivariate analysis, M7 AML was an independent adverse prognostic factor for OS, independent of other factors including cytogenetic abnormalities (hazard ratio 1.51, P = .049). These results confirm the poor prognosis of M7 AML and indicate that other biologic characteristics beyond cytogenetic abnormalities likely play a role in this disease.


Carcinoma ex pleomorphic adenoma with sebaceous differentiation in the mandibular salivary gland of a dog.
A carcinoma ex pleomorphic adenoma was diagnosed in the left mandibular salivary gland of an 8-year-old female spayed dog. The animal presented with a large nonpainful swelling in the left submandibular region. A computed tomography scan detected an irregularly enhancing soft tissue mass that was closely associated with the left external ear canal and extended to the left wing of the atlas. On surgical exploration, the mass was intimately associated with the left mandibular salivary gland. Both the mass and the adjacent gland were removed, and the diagnosis was determined by histopathology. The tumor was comprised of basaloid and low columnar epithelial cells, many glandular units formed by well-differentiated sebocytes, and multifocal regions of necrosis, mineralization, and hemorrhage. Salivary gland tumors with sebaceous differentiation are very rare in animals, with one previously reported case in a cat.


Differential expression of IL-17, 22 and 23 in the progression of colorectal cancer in patients with K-ras mutation: Ras signal inhibition and crosstalk with GM-CSF and IFN-γ.
Recent studies have suggested that aberrant K-ras signaling is responsible for triggering immunological responses and inflammation-driven tumorigenesis. Interleukins IL-17, IL-22, and IL-23 have been reported in various types of malignancies, but the exact mechanistic role of these molecules remains to be elucidated. Given the role of K-ras and the involvement of interleukins in colorectal tumorigenesis, research efforts are reported for the first time, showing that differentially expressed interleukin IL-17, IL-22, and IL-23 levels are associated with K-ras in a stage-specific fashion along colorectal cancer progression. Specifically, a) the effect of K-ras signaling was investigated in the overall expression of interleukins in patients with colorectal cancer and healthy controls, and b) an association was established between mutant K-ras and cytokines GM-CSF and IFN-γ. The results indicate that specific interleukins are differentially expressed in K-ras positive patients and the use of K-ras inhibitor Manumycin A decreases both interleukin levels and apoptosis in Caco-2 cells by inhibiting cell viability. Finally, inflammation-driven GM-CSF and IFN-γ levels are modulated through interleukin expression in tumor patients, with interleukin expression in the intestinal lumen and cancerous tissue mediated by aberrant K-ras signaling. Collectively, the findings a) indicate that interleukin expression is influenced by ras signaling and specific interleukins play an oncogenic promoter role in colorectal cancer, highlighting the molecular link between inflammation and tumorigenesis, and b) accentuate the interwoven molecular correlations as leads to new therapeutic approaches in the future.


Spine Trauma-What Are the Current Controversies?
Although less common than other musculoskeletal injuries, spinal trauma may lead to significantly more disability and costs. During the last 2 decades there was substantial improvement in our understanding of the basic patterns of spinal fractures leading to more reliable classification and injury severity assessment systems but also rapid developments in surgical techniques. Despite these advancements, there remain unresolved issues concerning the management of these injuries. At this moment there is persistent controversy within the spinal trauma community, which can be grouped under 6 headings. First of all there is still no unanimity on the role and timing of medical and surgical interventions for patients with associated neurologic injury. The same is also true for type and timing of surgical intervention in multiply injured patients. In some common injury types like odontoid fractures and burst type (A3-A4) fractures in thoracolumbar spine, there is wide variation in practice between operative versus nonoperative management without clear reasons. Also, the role of different surgical approaches and techniques in certain injury types are not clarified yet. Methods of nonoperative management and care of elderly patients with concurrent complex disorders are also areas where there is no consensus. In this overview article the main reasons for these controversies are reviewed and the possible ways for resolutions are discussed.


Xanthohumol, a Prenylated Flavonoid from Hops, Induces Caspase-Dependent Degradation of Oncoprotein BCR-ABL in K562 Cells.
BCR-ABL oncoprotein drives the initiation, promotion, and progression of chronic myelogenous leukemia (CML). Tyrosine kinase inhibitors are the first choice for CML therapy, however, BCR-ABL mediated drug resistance limits its clinical application and prognosis. A novel promising therapeutic strategy for CML therapy is to degrade BCR-ABL using small molecules. Antioxidant xanthohumol (XN) is a hop-derived prenylated flavonoid with multiple bioactivities. In this study, we showed XN could inhibit the proliferation, induce S phase cell cycle arrest, and stimulate apoptosis in K562 cells. XN degraded BCR-ABL in a concentration- and time-dependent manner, and the involved degradation pathway was caspase activation, while not autophagy induction or ubiquitin proteasome system (UPS) activation. Moreover, we revealed for the first time that XN could inhibit the UPS and autophagy in K562 cells, and the inhibitory effect of XN on autophagy could attenuate imatinib-induced autophagy and enhance the therapeutic efficiency of imatinib in K562 cells. Our present findings identified XN act as a degrader of BCR-ABL in K562 cells, and XN had potential to be developed as an alternate agent for CML therapy.


Hydrogen-assisted step-edge nucleation of MoSe2 monolayers on sapphire substrates.
The fabrication of large-area single crystalline monolayer transition metal dichalcogenides (TMDs) is essential for a range of electric and optoelectronic applications. Chemical vapor deposition (CVD) is a promising method to achieve this goal by employing orientation control or alignment along the crystalline lattice of the substrate such as sapphire. On the other hand, a fundamental understanding of the aligned-growth mechanism of TMDs is limited. In this report, we show that the controlled introduction of H2 during the CVD growth of MoSe2 plays a vital role in the step-edge aligned nucleation on a c-sapphire (0001) substrate. In particular, the MoSe2 domains nucleate along the [112[combining macron]0] step-edge orientation by flowing H2 subsequent to pure Ar. Systematic studies, including the H2 introduction time, flow rate, and substrate temperature, suggest that the step-edge aligned nucleation of MoSe2 can be controlled by the hydrogen concentration on the sapphire substrate. These results offer important insights into controlling the epitaxial growth of 2D materials on a crystalline substrate.


Valency of HER2 Targeting Antibodies Influences Tumor Cell Internalization and Penetration.
T-cell-dependent bispecific antibodies (TDB) have been a major advancement in the treatment of cancer, allowing for improved targeting and efficacy for large molecule therapeutics. TDBs are comprised of one arm targeting a surface antigen on a cancer cell and another targeting an engaging surface antigen on a cytotoxic T cell. To impart this function, the antibody must be in a bispecific format as opposed to the more conventional bivalent format. Through in vitro and in vivo studies, we sought to determine the impact of changing antibody valency on solid tumor distribution and catabolism. A bivalent anti-HER2 antibody exhibited higher catabolism than its full-length monovalent binding counterpart in vivo by both invasive tissue harvesting and noninvasive single photon emission computed tomography/X-ray computed tomography imaging despite similar systemic exposures for the two molecules. To determine what molecular factors drove in vivo distribution and uptake, we developed a mechanistic model for binding and catabolism of monovalent and bivalent HER2 antibodies in KPL4 cells. This model suggests that observed differences in cellular uptake of monovalent and bivalent antibodies are caused by the change in apparent affinity conferred by avidity as well as differences in internalization and degradation rates of receptor bound antibodies. To our knowledge, this is the first study to directly compare the targeting abilities of monovalent and bivalent full-length antibodies. These findings may inform diverse antibody therapeutic modalities, including T-cell-redirecting therapies and drug delivery strategies relying upon receptor internalization.


Resection margins in carcinoma of the head of the pancreas. Implications for radiation therapy.
A retrospective review of the pathology and clinical course of 72 patients undergoing resection of carcinoma of the head of the pancreas was undertaken to identify the frequency of tumor involvement at standard surgical transection margins (stomach, duodenum, pancreas, and bile duct) as well as the peripancreatic soft tissue margin and the potential clinical significance of these findings. Of 72 patients undergoing resection, 37 patients (51%) were found to have tumor extension to the surgical margins. The most commonly involved margin was peripancreatic soft tissue (27 patients) followed by pancreatic transection line (14 patients) and bile duct transection line (4 patients). For 37 patients with tumor present at a resection margin, there were no survivors beyond 41 months. No difference in survival or local control was seen between 14 patients receiving postoperative radiation therapy and 5-fluorouracil (5-FU) compared with 23 patients not receiving additional treatment. In contrast, the 5-year actuarial survival and local control of 35 patients undergoing resection without tumor invasion to a resection margin was 22% and 43%, respectively. The 5-year survival and local control of 16 patients receiving adjuvant radiation therapy and 5-FU was 29% and 42%, respectively, whereas these figures were 18% and 31% for 19 patients not receiving adjuvant therapy (p > 0.10). Because residual local tumor after resection is common, preoperative radiation therapy may be beneficial in this disease. It should minimize the risk of dissemination during operative manipulation and facilitate a curative resection by promoting tumor regression. Because local failure rates approach 60% after resection and adjuvant therapy even in cases having clear resection margins, intraoperative radiation therapy to the tumor bed at the time of resection also might be considered. Protocols evaluating the feasibility and efficacy of preoperative radiation therapy and resection with intraoperative radiation therapy for patients with pancreatic cancer are underway.


Risk factors and clinical features of deterioration in COVID-19 patients in Zhejiang, China: a single-centre, retrospective study.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection swept through Wuhan and spread across China and overseas beginning in December 2019. To identify predictors associated with disease progression, we evaluated clinical risk factors for exacerbation of SARS-CoV-2 infection. A retrospective analysis was used for PCR-confirmed COVID-19 (coronavirus disease 2019)-diagnosed hospitalized cases between January 19, 2020, and February 19, 2020, in Zhejiang, China. We systematically analysed the clinical characteristics of the patients and predictors of clinical deterioration. One hundred patients with COVID-19, with a median age of 54 years, were included. Among them, 49 patients (49%) had severe and critical disease. Age ([36-58] vs [51-70], P = 0.0001); sex (49% vs 77.6%, P = 0.0031); Body Mass Index (BMI) ([21.53-25.51] vs [23.28-27.01], P = 0.0339); hypertension (17.6% vs 57.1%, P < 0.0001); IL-6 ([6.42-30.46] vs [16.2-81.71], P = 0.0001); IL-10 ([2.16-5.82] vs [4.35-9.63], P < 0.0001); T lymphocyte count ([305-1178] vs [167.5-440], P = 0.0001); B lymphocyte count ([91-213] vs [54.5-163.5], P = 0.0001); white blood cell count ([3.9-7.6] vs [5.5-13.6], P = 0.0002); D2 dimer ([172-836] vs [408-953], P = 0.005), PCT ([0.03-0.07] vs [0.04-0.15], P = 0.0039); CRP ([3.8-27.9] vs [17.3-58.9], P < 0.0001); AST ([16, 29] vs [18, 42], P = 0.0484); artificial liver therapy (2% vs 16.3%, P = 0.0148); and glucocorticoid therapy (64.7% vs 98%, P < 0.0001) were associated with the severity of the disease. Age and weight were independent risk factors for disease severity. Deterioration among COVID-19-infected patients occurred rapidly after hospital admission. In our cohort, we found that multiple factors were associated with the severity of COVID19. Early detection and monitoring of these indicators may reduce the progression of the disease. Removing these factors may halt the progression of the disease. In addition, Oxygen support, early treatment with low doses of glucocorticoids and artificial liver therapy, when necessary, may help reduce mortality in critically ill patients.


African Americans with a family history of colorectal cancer: barriers and facilitators to screening.
To explore barriers and facilitators of screening for colorectal cancer (CRC), as well as suggestions for improving screening among African Americans with first-degree relatives with CRC. A qualitative, descriptive approach involving focus groups. A community healthcare clinic in Baltimore, MD. 14 African American men and women aged 40 or older with at least one first-degree family member affected by CRC. In-depth focus groups were conducted until thematic saturation was achieved. Thematic analysis and data reduction were conducted using ATLAS.ti, version 5.0. CRC screening barriers and facilitators. The participants were mostly male, insured, and had a parent with CRC. Commonly reported barriers to CRC screening included fear of serious illness, mistrust of the medical establishment, potential screening discomfort, lack of information on CRC risk factors, lack of healthcare access, absence of symptoms, no knowledge of CRC screening benefits, community reticence about cancer, and CRC myths. Facilitating factors for CRC screening included a belief of personal risk for CRC, physician recommendations, and acknowledgment of age as a risk factor. Suggestions to increase screening rates included distribution of culturally appropriate and community-based efforts (e.g., mobile units, church-based interventions). Participants also suggested ways to increase motivation and provide social support for screening patients. Additional research is needed to identify and test effective screening approaches for this underserved group at increased risk for CRC. Study results suggest that cancer risk and screening education, coupled with screening opportunities in the community, may yield increased screening rates. Lack of knowledge about CRC and CRC screening exists in the study population. Promoting screening across generations, developing and disseminating culturally appropriate educational materials within the community, and encouraging older individuals to screen to take care of their family may be appropriate interventions.


Strategies of survival and resource exploitation in the Antarctic fellfield ecosystem.
Antarctic fellfields present organisms with a heterogeneous habitat characterised by a wide variety of environmental stresses. These include low temperatures, limited moisture availability, frequent and often rapid freeze-thaw and hydration-dehydration cycles, exposure to high photosynthetic photon flux density and ultraviolet (uv) irradiance, seasonal snow cover, high winds, cryoturbation and, depending on location south of the Antarctic Circle, considerable daylight in summer. Most of these factors vary both predictably and unpredictably in spatial and temporal planes. In response to this adverse environment, fellfield organisms have developed a variety of strategies to overcome physiological stress and to exploit the limited resources available during the short austral growing season. A high degree of synchronisation exists, so that investment in non-essential activity and adaptations is minimised. Here, we review the combined suites of co-adapted traits used by different fellfield taxa to achieve energy acquisition, growth and reproduction under adverse levels of two principal limiting factors: low temperatures and the scarcity of water. To this end, a detailed characterisation of the Antarctic fellfield microenvironment is followed by a synthesis of available data on the morphology, physiology, life history and behaviour of successful Antarctic flora and fauna. Tolerance of low temperatures by fellfield organisms is achieved by elevation of standard metabolism, production and accumulation of cryoprotectants, supercooling, melanic pigmentation, behavioural avoidance, compact growth forms and synchronised reproduction and extended life cycles. Low moisture conditions are overcome by dehydration resistance, anhydrobiosis, development of resting stages and by behavioural avoidance of desiccating conditions. Occupancy of the Antarctic fellfield habitat is considered to require the ability to respond rapidly to ephemeral resources and to tolerate severe environmental stresses. During summer, organisms rely on opportunism to maintain a positive energy balance. During winter, resistance adaptations are used to withstand the potentially lethal climate, especially in habitats not protected by snow cover. This deterministic framework has led to the selection of species that are genetically and physiologically pre-adapted for resource acquisition yet sufficiently robust to withstand cold and desiccation stresses. Non-adapted taxa fail to become established. Despite the environmental selection pressures, available evidence suggests that colonisation of the fellfield habitat has not required the evolution of any adaptations, only the refinement of those already possessed to an extent by some temperate forms. This has led to the convergence of survival strategies. It is hypothesised that, in the short term, the majority of Antarctic fellfield biota are able to absorb the predicted effects of a changing climate by their high levels of physiological tolerance and life-cycle flexibility.


Dual Langmuir-probe array for 3D plasma studies in TORPEX.
We have designed and installed a new Langmuir-probe (LP) array diagnostic to determine basic three-dimensional (3D) features of plasmas in TORPEX. The diagnostic consists of two identical LP arrays, placed on opposite sides of the apparatus, which provide comprehensive coverage of the poloidal cross section at the two different toroidal locations. Cross correlation studies of signals from the arrays provide a basic way to extract 3D information from the plasmas, as experiments show. Moreover, the remarkable signal-to-noise performance of the front-end electronics allows us to follow a different approach in which we combine information from all probes in both arrays to reconstruct elementary 3D plasma structures at each acquisition time step. Then, through data analysis, we track the structures as they evolve in time. The LP arrays include a linear-motion mechanism that can displace radially the probes located on the low field side for experiments that require fine-tuning of the probe locations, and for operational compatibility with the recently installed in-vessel toroidal conductor.


MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus.
Prolonged seizures (status epilepticus, SE) may drive hippocampal dysfunction and epileptogenesis, at least partly, through an elevation in neurogenesis, dysregulation of migration and aberrant dendritic arborization of newly-formed neurons. MicroRNA-22 was recently found to protect against the development of epileptic foci, but the mechanisms remain incompletely understood. Here, we investigated the contribution of microRNA-22 to SE-induced aberrant adult neurogenesis. SE was induced by intraamygdala microinjection of kainic acid (KA) to model unilateral hippocampal neuropathology in mice. MicroRNA-22 expression was suppressed using specific oligonucleotide inhibitors (antagomir-22) and newly-formed neurons were visualized using the thymidine analog iodo-deoxyuridine (IdU) and a green fluorescent protein (GFP)-expressing retrovirus to visualize the dendritic tree and synaptic spines. Using this approach, we quantified differences in the rate of neurogenesis and migration, the structure of the apical dendritic tree and density and morphology of dendritic spines in newly-formed neurons.SE resulted in an increased rate of hippocampal neurogenesis, including within the undamaged contralateral dentate gyrus (DG). Newly-formed neurons underwent aberrant migration, both within the granule cell layer and into ectopic sites. Inhibition of microRNA-22 exacerbated these changes. The dendritic diameter and the density and average volume of dendritic spines were unaffected by SE, but these parameters were all elevated in mice in which microRNA-22 was suppressed. MicroRNA-22 inhibition also reduced the length and complexity of the dendritic tree, independently of SE. These data indicate that microRNA-22 is an important regulator of morphogenesis of newly-formed neurons in adults and plays a role in supressing aberrant neurogenesis associated with SE.


Quantification of pravastatin acid, lactone and isomers in human plasma by UHPLC-MS/MS and its application to a pediatric pharmacokinetic study.
An ultra high pressure liquid chromatography-tandem mass spectrometric (UHPLC-MS/MS) method for the simultaneous quantitation of pravastatin and major metabolites, 3'α-hydroxy-pravastatin, pravalactone and 3'α-hydroxy-pravalactone, in human plasma has been developed and validated. Aliquots of (100μL) plasma in EDTA were diluted in pH 4.5 (0.1M buffer) to stabilize the analytes and subjected to hydrophilic lipophilic balance (HLB) solid phase extraction on 96 well μelution plates. Extracted samples were evaporated to dryness and reconstituted in pH 4.5 buffer. Chromatographic separation was performed on a Cortecs™ C18 column (2.1×100mm, 1.8μm), using gradient elution with a blend of acetonitrile and 10mM methylammonium acetate buffer (pH 4.5) at a flow rate of 0.4mL/min. Mass spectrometric detection was performed using multiple reaction monitoring (MRM) switching between positive/negative electrospay ionization (ESI). Pravastatin, 3'α-hydroxy-pravastatin, and internal standards [(2)H3]-pravastatin, and [(2)H3]-3'α-hydroxy-pravastatin were monitored in negative ESI mode at ion transitions m/z 423.2→321.1 and 426.2→321.1, respectively. Positive ESI mode was used for the detection of pravalactone, 3'α-hydroxy-pravalactone, and internal standards [(2)H3]-pravalactone, and [(2)H3]-3'α-hydroxy-pravalactone at ion transitions m/z 438.2→183.1 and 441.2→269.1 respectively. The method was linear for all analytes in the concentration range 0.5-200nM with intra- and inter-day precisions (as relative standard deviation) of ≤5.2% and accuracy (as relative error) of ≤8.0% at all quality control levels. The method was successfully applied to the investigation of pharmacokinetic properties of pravastatin and its metabolites in children after an oral dose of 20-40mg.


The response of the lamb ductus arteriosus to endothelin: developmental changes and influence of light.
Endothelin-1 (ET-1) is a putative messenger of oxygen in the ductus arteriosus. Since the ability of the vessel to contract to oxygen increases with gestation, we wished to ascertain whether ET-1 action is also developmentally regulated. A corollary objective was to assess whether any gestational variation in the ET-1 contraction is due to a change in the ET(A)-mediated action or to a shift in the balance between opposing, contractile (ET(A) - mediated) and relaxant (ET(B)-mediated), actions. Experiments were performed with isolated ductal strips from preterm (0.7 gestation) and near-term fetal lambs. ET-1 contracted the ductus dose-dependently (10(-10)-10(-7) M) at both ages; however, the peak contraction was about double in magnitude at term. Regardless of age, ET-1 contraction was greater with preparations kept in the dark compared to those exposed to light. This effect of light was not seen after removing the endothelium or when treating the intact tissue with the ET(B) antagonist BQ788 (1 microM). In the dark, however, BQ788 did not modify significantly the ET-1 response at either age. We conclude that ET-1 becomes a stronger ductus constrictor with fetal age, conceivably by acting on ET(A) receptors. Hence, the concept of ET-1 mediating the oxygen contraction is further validated. Peculiarly, the ET-1 contraction is curtailed by light through a hitherto undefined ET(B) receptor-linked process.


An overview of lymphatic filariasis lymphedema.
Filariasis is caused by thread-like nematode worms and is classified according to their presence in the vertebrate host. The lymphatic group includes Wuchereria bancrofti, Brugia malayi, and Brugia timori. Lymphatic filariasis, a mosquito-borne disease, has been one of the most prevalent diseases in tropical and subtropical countries and is accompanied by a number of pathological conditions. It is estimated that currently (after 13 years of the MDA programme) there are an estimated 67.88 million LF cases that include 36.45 million microfilaria carriers, 19.43 million hydrocele cases, and 16.68 million lymphedema cases. Adult filarial worms reside in the lymphatics and lymph nodes and induce changes that result in dilatation of lymphatics and thickening of the lymphatic vessel walls. Progressive lymphatic damage and pathology results from the summation of the effect of tissue alterations induced by both living and nonliving adult parasites. In recent years, there has been rapid progress in filariasis research, which has provided new insights into the pathogenesis of filarial disease, diagnosis, chemotherapy, the host-parasite relationship, and the genomics of the parasite. We examined the clinical manifestations of the disease, diagnosis, treatment, immune responses, and management including review of pharmaceutical agents against filariasis. Details on infection, safety profile, and status in clinical practices are also reported.


Detection and typing of human papillomaviruses combining different methods: polymerase chain reaction, restriction fragment length polymorphism, line probe assay and sequencing.
The identification of the etiological factor of many cervical precancerous lesions and cervical cancer, the human papillomavirus (HPV) is widely used. In this study, we evaluated the consensus and type-specific (TS) polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), line probe assay (LiPA, Innogenetics) and sequencing to determine the HPV types in cervical specimens. Out of 690 High-grade Squamous Intraepithelial Lesion (HSIL) samples, 86.7% were HPV positive and 13.3% HPV negative by consensus primers (MY09/MY11, L1C1/L1C2-1/L1C2-2 and/or GP5/6) directed PCR. Out of 598 HPV positive samples, 85.3% were typed by TS-PCR being HPV 6/11, 16, 18, 31 and/or 33, while 14.7% remained untyped. The most prevalent HPV type in the study group was HPV 16, identified in 35.5% cases, while HPV 31 was the second most frequent HPV type with a prevalence of 10.5%. They were followed by HPV types 6/11, 33 and 18 with a prevalence of 7.4%, 6.2% and 4.9%, respectively. Multiple HPV infections with two or more HPV types (6/11, 16, 18, 31 and/or 33) were found in 9.4% cases. A subset of 88 samples was further typed by RFLP and LiPA to determine the rare HPV types in HSIL samples. The most frequent low abundant HPV types in single infections in decreasing order were HPV 53, 58, 66, 56 and 52, while HPV 51 was the most frequent low abundant HPV type found in multiple HPV infections. Multiple HPV infections were mostly found by LiPA in 27.3% cases versus 14.8% cases found by RFLP. The perfect agreement between RFLP and LiPA assay pair was observed only for HPV types 16, 18, 34 and 59 (kappa value of 1). For other HPV types, the inter-assay agreement ranged from very good to no agreement indicating that neither assay is perfect. Sequencing was performed for 33 samples in cases where both RFLP and LiPA were inconclusive. Sequencing was shown to be a very good method in case of single HPV infection but not applicable in case of multiple HPV infections. Both RFLP and LiPA are good assays for epidemiological studies, although RFLP being cumbersome and time-consuming is less applicable than LiPA. Careful consideration has to be made before the implementation of either HPV typing methods in clinical laboratories.


Transcatheter closure of ruptured sinus of valsalva aneurysm.
Percutaneous transcatheter closure of ruptured sinus of valsalva aneurysm was attempted in eight patients between January 1995 and March 2003 as an alternative strategy to surgery as this technique at present is an accepted therapeutic modality for various intracardiac defects. The age range was 14-35 years, all were male, seven in symptomatic class III and one in class IV on medical treatment. Two-dimensional and color Doppler echocardiography revealed rupture of an aneurysm of right coronary sinus into right ventricle in five and noncoronary sinus into right atrium in three and none had associated ventricular septal defect. The echo estimated size of the defect was 7-12 mm. On cardiac catheterization left ventricular end-diastolic pressure ranged from 20 to 40 mmHg and the calculated Qp/Qs ratio was 2-3.5. In all patients the defect was crossed retrogradely from the aortic side and over an arterio-venous wire loop after balloon sizing, devices were successfully deployed by antegrade venous approach (Rashkind umbrella device in two and Amplatzer occluders in six, which included Amplatzer duct occluder in five and Amplatzer septal occluder in one). One patient who had residual shunt developed hemolysis on the next day and was taken up for reintervention. That patient continued to have intermittent hemolysis and was sent for surgical repair. On follow-up (2-96 months), there was no device embolization, infective endocarditis, and aortic regurgitation. One patient died of progressive congestive heart failure while other six are asymptomatic. These data highlight that transcatheter closure is feasible and effective, especially safe with the available Amplatzer devices. Definitely, it has the advantage of obviating open heart surgery but complete occlusion is mandatory to prevent hemolysis and infective endocarditis.


Quantitative sequence-function relationships in proteins based on gene ontology.
The relationship between divergence of amino-acid sequence and divergence of function among homologous proteins is complex. The assumption that homologs share function--the basis of transfer of annotations in databases--must therefore be regarded with caution. Here, we present a quantitative study of sequence and function divergence, based on the Gene Ontology classification of function. We determined the relationship between sequence divergence and function divergence in 6828 protein families from the PFAM database. Within families there is a broad range of sequence similarity from very closely related proteins--for instance, orthologs in different mammals--to very distantly-related proteins at the limit of reliable recognition of homology. We correlated the divergence in sequences determined from pairwise alignments, and the divergence in function determined by path lengths in the Gene Ontology graph, taking into account the fact that many proteins have multiple functions. Our results show that, among homologous proteins, the proportion of divergent functions decreases dramatically above a threshold of sequence similarity at about 50% residue identity. For proteins with more than 50% residue identity, transfer of annotation between homologs will lead to an erroneous attribution with a totally dissimilar function in fewer than 6% of cases. This means that for very similar proteins (about 50 % identical residues) the chance of completely incorrect annotation is low; however, because of the phenomenon of recruitment, it is still non-zero. Our results describe general features of the evolution of protein function, and serve as a guide to the reliability of annotation transfer, based on the closeness of the relationship between a new protein and its nearest annotated relative.


Impaired learning in adulthood following neonatal delta9-THC exposure.
Cannabis is one of the most commonly used illicit drugs during pregnancy, but little is known about the lasting effects of early-life exposure to this drug. In this study, male Wistar rat pups were treated daily with (-)-delta9-tetrahydrocannabinol (THC; 5 mg/kg, s.c.) or its vehicle between postnatal days (PND) 4 and 14. Drug administration during this early postnatal period in rats is analogous to the third trimester of gestation in humans, which is a major period of synaptogenesis. Rats were subsequently tested drug-free during young adulthood (PND 56) using a two-component food-motivated double Y-maze test. Each trial included distinct spatial discrimination and delayed alternation components, which permitted the simultaneous assessment of reference memory and working memory. Rats were tested for 30 trials/day, 5 days/week for 5 weeks. Results revealed no significant differences between THC- and vehicle-treated rats in the spatial discrimination task. However, compared to vehicle-treated rats, THC-treated rats committed significantly more errors, and required significantly longer to obtain 80% correct over two consecutive days in the delayed alternation task. Results suggest that neonatal THC exposure leads to a specific and lasting deficit in learning in adulthood, which is likely due to impaired working memory function.


Dual impact of elevated temperature on plant defence and bacterial virulence in Arabidopsis.
Environmental conditions profoundly affect plant disease development; however, the underlying molecular bases are not well understood. Here we show that elevated temperature significantly increases the susceptibility of Arabidopsis to Pseudomonas syringae pv. tomato (Pst) DC3000 independently of the phyB/PIF thermosensing pathway. Instead, elevated temperature promotes translocation of bacterial effector proteins into plant cells and causes a loss of ICS1-mediated salicylic acid (SA) biosynthesis. Global transcriptome analysis reveals a major temperature-sensitive node of SA signalling, impacting ~60% of benzothiadiazole (BTH)-regulated genes, including ICS1 and the canonical SA marker gene, PR1. Remarkably, BTH can effectively protect Arabidopsis against Pst DC3000 infection at elevated temperature despite the lack of ICS1 and PR1 expression. Our results highlight the broad impact of a major climate condition on the enigmatic molecular interplay between temperature, SA defence and function of a central bacterial virulence system in the context of a widely studied susceptible plant-pathogen interaction.


Cost-assessment of alternative management strategies for achalasia.
Achalasia is a primary oesophageal motor disorder characterised by the abnormal relaxation of the lower oesophageal sphincter (LES) and absent oesophageal peristalsis. It is a rare disease, with an estimated incidence of approximately 1/100,000 and a prevalence close to 10/100,000 [1]. Its exact aetiology remains unknown. Autoimmune, infectious, degenerative and hereditary processes have all been proposed as factors that lead to a chronic inflammatory response in the myenteric plexus, thus resulting in selective loss of inhibitory neurons [2] and failure of the LES to relax and aperistalsis in the body of the oesophagus. The most common symptoms of achalasia are dysphagia for solids and liquids, regurgitation, chest pain, weight loss and heartburn in > 90 approximately 75, 40 - 50, approximately 60, approximately 40%, respectively [3,4]. The diagnosis is based on symptoms, barium swallow and manometry. A barium oesophagram typically shows a dilated oesophagus that tapers into a 'bird-beak' at the gastro-oesophageal junction with lack of normal peristalsis on fluoroscopic evaluation. The characteristic manometric features of achalasia are abnormal LES relaxation and aperistalsis; additionally, the LES pressure is frequently high, but can also be normal. Current practice of medicine is faced with rising healthcare costs and limited budgets [5]. We are therefore confronted with an increasing demand to justify the value of our therapeutic interventions, not only from the risk/benefit standpoint but also from the cost perspective [6,7].


Blue and Red Light-Evoked Pupil Responses in Photophobic Subjects with TBI.
Photophobia is a common symptom in individuals suffering from traumatic brain injury (TBI). Recent evidence has implicated blue light-sensitive intrinsically photosensitive retinal ganglion cells (ipRGCs) in contributing to the neural circuitry mediating photophobia in migraine sufferers. The goal of this work is to test the hypothesis that ipRGC function is altered in TBI patients with photophobia by assessing pupillary responses to blue and red light. Twenty-four case participants (mean age 43.3; 58% female), with mild TBI and self-reported photophobia, and 12 control participants (mean age 42.6; 58% female) were in this study. After 10 minutes of dark adaptation, blue (470 nm, 1 × 10 phots/s/cm) and red (625 nm, 7 × 10 phots/s/cm) flashing (0.1 Hz) light stimuli were delivered for 30 seconds to the dilated left eye while the right pupil was recorded. The amplitude of normalized pupil fluctuation (constriction and dilation) was quantified using Fourier fast transforms. In both case and control participants, the amplitude of pupil fluctuation was significantly less for the blue light stimuli as compared to the red light stimuli, consistent with a contribution of ipRGCs to these pupil responses. There was no significant difference in the mean pupil fluctuation amplitudes between the two participant groups, but case participants displayed greater variability in their pupil responses to the blue stimulus. Case and control participants showed robust ipRGC-mediated components in their pupil responses to blue light. The results did not support the hypothesis that ipRGCs are "hypersensitive" to light in TBI participants with photophobia. However, greater pupil response variability in the case subjects suggests that ipRGC function may be more heterogeneous in this group.


Budget impact analysis: review of the state of the art.
A budget impact analysis for a new pharmaceutical product provides estimates of the likely impact of the new drug on a healthcare decision maker's short- and longer-term annual budgets. Budget impact analysis are an essential part of a comprehensive economic assessment of a new pharmaceutical product and are increasingly required, along with cost-effectiveness analyses, before national or local formulary approval or reimbursement. Standards for the development, content and presentation of the results of a budget impact analysis have recently been proposed but detailed guidance on alternative methods for estimating budget impacts are not yet available. Methods for performing these analyses require either static or dynamic analysis techniques depending on the type of health condition and health impact of the new drug. Several factors, that are not generally needed for cost-effectiveness analysis, should be part of a comprehensive budget impact analysis including the size of the treated population, second-order costs, market diffusion rates for the new drug, and off-label use. A review of the recent literature indicates that there is only a limited number of published budget impact analyses and these vary greatly in the methods used. Instead of publication in peer-reviewed journals, budget impact analyses are more frequently presented directly to decision makers as interactive computer programs designed to compute the budget impact for specific health plans using plan-specific inputs. It is recommend that a comprehensive approach to budget impact estimation be adopted, with the results being presented from both a societal perspective as well as from more limited perspectives depending on the needs of the decision maker.


Pulsed interleaved excitation fluctuation imaging.
Fluorescence fluctuation imaging is a powerful means to investigate dynamics, interactions, and stoichiometry of proteins inside living cells. Pulsed interleaved excitation (PIE) is the method of nanosecond alternating excitation with time-resolved detection and allows accurate, independent, and quasi-simultaneous determination of fluorescence intensities and lifetimes of different fluorophores. In this work, we combine pulsed interleaved excitation with fluctuation imaging methods (PIE-FI) such as raster image correlation spectroscopy (RICS) or number and brightness analysis (N&B). More specifically, we show that quantitative measurements of diffusion and molecular brightness of Venus fluorescent protein (FP) can be performed in solution with PIE-RICS and compare PIE-RICS with single-point PIE-FCS measurements. We discuss the advantages of cross-talk free dual-color PIE-RICS and illustrate its proficiency by quantitatively comparing two commonly used FP pairs for dual-color microscopy, eGFP/mCherry and mVenus/mCherry. For N&B analysis, we implement dead-time correction to the PIE-FI data analysis to allow accurate molecular brightness determination with PIE-NB. We then use PIE-NB to investigate the effect of eGFP tandem oligomerization on the intracellular maturation efficiency of the fluorophore. Finally, we explore the possibilities of using the available fluorescence lifetime information in PIE-FI experiments. We perform lifetime-based weighting of confocal images, allowing us to quantitatively determine molecular concentrations from 100 nM down to <30 pM with PIE-raster lifetime image correlation spectroscopy (RLICS). We use the fluorescence lifetime information to perform a robust dual-color lifetime-based FRET analysis of tandem fluorescent protein dimers. Lastly, we investigate the use of dual-color RLICS to resolve codiffusing FRET species from non-FRET species in cells. The enhanced capabilities and quantitative results provided by PIE-FI make it a powerful method that is broadly applicable to a large number of interesting biophysical studies.


Evidence for redox sensing by a human cardiac calcium channel.
Ion channels are critical to life and respond rapidly to stimuli to evoke physiological responses. Calcium influx into heart muscle occurs through the ion conducting α1C subunit (Cav1.2) of the L-type Ca(2+) channel. Glutathionylation of Cav1.2 results in increased calcium influx and is evident in ischemic human heart. However controversy exists as to whether direct modification of Cav1.2 is responsible for altered function. We directly assessed the function of purified human Cav1.2 in proteoliposomes. Truncation of the C terminus and mutation of cysteines in the N terminal region and cytoplasmic loop III-IV linker did not alter the effects of thiol modifying agents on open probability of the channel. However mutation of cysteines in cytoplasmic loop I-II linker altered open probability and protein folding assessed by thermal shift assay. We find that C543 confers sensitivity of Cav1.2 to oxidative stress and is sufficient to modify channel function and posttranslational folding. Our data provide direct evidence for the calcium channel as a redox sensor that facilitates rapid physiological responses.


Inhibition of porcine liver carboxylesterase by a new flavone glucoside isolated from Deverra scoparia.
An endemic North African Saharan plant from of the Apiaceae family, Deverra scoparia, used locally for medicinal preparations, showed a strong inhibitory effect on porcine liver carboxylesterase. The active compound from the aerial part of the plant was purified by semi-preparative HPLC and photodiode array detection, and structurally determined by (1)H, (13)C NMR and mass spectroscopy methods. This compound was identified as flavone-3,4',7-trihydroxy-3'-methoxy-7-glucoside and it was found to be a powerful competitive inhibitor of porcine liver carboxylesterase with a inhibition constant value of 16 microM. Based on the structural features of the inhibitor and the enzyme active site region, it seems that the flavonoside binds to the surface of the enzyme. The low K(i) value suggests some physiological significance of such inhibitory activity, especially concerning the bio-transformation of xenobiotics.


Protein kinase C δ regulates the release of collagen type I from vascular smooth muscle cells via regulation of Cdc42.
Collagen type I is the most abundant component of extracellular matrix in the arterial wall. Mice knocked out for the protein kinase C δ gene (PKCδ KO) show a marked reduction of collagen I in the arterial wall. The lack of PKCδ diminished the ability of arterial smooth muscle cells (SMCs) to secrete collagen I without significantly altering the intracellular collagen content. Moreover, the unsecreted collagen I molecules accumulate in large perinuclear puncta. These perinuclear structures colocalize with the trans-Golgi network (TGN) marker TGN38 and to a lesser degree with cis-Golgi marker (GM130) but not with early endosomal marker (EEA1). Associated with diminished collagen I secretion, PKCδ KO SMCs exhibit a significant reduction in levels of cell division cycle 42 (Cdc42) protein and mRNA. Restoring PKCδ expression partially rescues Cdc42 expression and collagen I secretion in PKCδ KO SMCs. Inhibition of Cdc42 expression or activity with small interfering RNA or secramine A in PKCδ WT SMCs eliminates collagen I secretion. Conversely, restoring Cdc42 expression in PKCδ KO SMCs enables collagen I secretion. Taken together, our data demonstrate that PKCδ mediates collagen I secretion from SMCs, likely through a Cdc42-dependent mechanism.


Involvement of the beta-endorphin neurons of the hypothalamic arcuate nucleus in ethanol-induced place preference conditioning in mice.
Increasing evidence indicates that mu- and delta-opioid receptors are decisively involved in the retrieval of memories underlying conditioned effects of ethanol. The precise mechanism by which these receptors participate in such effects remains unclear. Given the important role of the proopiomelanocortin (POMc)-derived opioid peptide beta-endorphin, an endogenous mu- and delta-opioid receptor agonist, in some of the behavioral effects of ethanol, we hypothesized that beta-endorphin would also be involved in ethanol conditioning. In this study, we treated female Swiss mice with estradiol valerate (EV), which induces a neurotoxic lesion of the beta-endorphin neurons of the hypothalamic arcuate nucleus (ArcN). These mice were compared to saline-treated controls to investigate the role of beta-endorphin in the acquisition, extinction, and reinstatement of ethanol (0 or 2 g/kg; intraperitoneally)-induced conditioned place preference (CPP). Immunohistochemical analyses confirmed a decreased number of POMc-containing neurons of the ArcN with EV treatment. EV did not affect the acquisition or reinstatement of ethanol-induced CPP, but facilitated its extinction. Behavioral sensitization to ethanol, seen during the conditioning days, was not present in EV-treated animals. The present data suggest that ArcN beta-endorphins are involved in the retrieval of conditioned memories of ethanol and are implicated in the processes that underlie extinction of ethanol-cue associations. Results also reveal a dissociated neurobiology supporting behavioral sensitization to ethanol and its conditioning properties, as a beta-endorphin deficit affected sensitization to ethanol, while leaving acquisition and reinstatement of ethanol-induced CPP unaffected.


A mathematical model of the impact of present and future malaria vaccines.
With the encouraging advent of new malaria vaccine candidates, mathematical modelling of expected impacts of present and future vaccines as part of multi-intervention strategies is especially relevant. The impact of potential malaria vaccines is presented utilizing the EMOD model, a comprehensive model of the vector life cycle coupled to a detailed mechanistic representation of intra-host parasite and immune dynamics. Values of baseline transmission and vector feeding behaviour parameters are identified, for which local elimination is enabled by layering pre-erythrocytic vaccines of various efficacies on top of high and sustained insecticide-treated net coverage. The expected reduction in clinical cases is further explored in a scenario that targets children by adding a pre-erythrocytic vaccine to the EPI programme for newborns. At high transmission, there is a minimal reduction in clinical disease cases, as the time to infection is only slightly delayed. At lower transmission, there is an accelerating community-level protection that has subtle dependences on heterogeneities in vector behaviour, ecology, and intervention coverage. At very low transmission, the trend reverses as many children are vaccinated to prevent few cases. The maximum-impact setting is one in which the impact of increasing bed net coverage has saturated, vector feeding is primarily outdoors, and transmission is just above the threshold where small perturbations from a vaccine intervention result in large community benefits.


Bonding durability of photocured phenyl-P in TEGDMA to smear layer-retained bovine dentin.
The long-term durability of a photocured resin bond to ground bovine dentin with its smear layer retained was investigated. The bonding resin that was employed was composed of 5.0 wt% 2-(methacryloxy)ethyl phenyl hydrogen phosphate (phenyl-P) as a diffusion-promoting monomer, 0.5 wt% camphorquinone as a photosensitizer, and 0.5 wt% N-phenylglycine as a reducing agent in triethyleneglycol dimethacrylate. Following polymerization of the bonding resin, a composite resin was placed and photocured for 60 seconds. Prepared specimens were stored in 37 degrees C water for 1 day, 6 months, and 1 year. Measured tensile bond strengths were 6.7 MPa after 1 day of storage, 4.1 MPa after 6 months of storage, and 2.8 mPa after 1 year of storage. Examinations of the interfaces after fracture under tensile loading suggested that long-term water-immersion weakened the bonds between the photocured adhesive resin and the smear layer-retained dentin because there was insufficient diffusion of the adhesive resin through the retained smear layer.


The glucose signaling network in yeast.
Most cells possess a sophisticated mechanism for sensing glucose and responding to it appropriately. Glucose sensing and signaling in the budding yeast Saccharomyces cerevisiae represent an important paradigm for understanding how extracellular signals lead to changes in the gene expression program in eukaryotes. This review focuses on the yeast glucose sensing and signaling pathways that operate in a highly regulated and cooperative manner to bring about glucose-induction of HXT gene expression. The yeast cells possess a family of glucose transporters (HXTs), with different kinetic properties. They employ three major glucose signaling pathways-Rgt2/Snf3, AMPK, and cAMP-PKA-to express only those transporters best suited for the amounts of glucose available. We discuss the current understanding of how these pathways are integrated into a regulatory network to ensure efficient uptake and utilization of glucose. Elucidating the role of multiple glucose signals and pathways involved in glucose uptake and metabolism in yeast may reveal the molecular basis of glucose homeostasis in humans, especially under pathological conditions, such as hyperglycemia in diabetics and the elevated rate of glycolysis observed in many solid tumors.


Cholelithiasis in adult bearded dragons: retrospective study of nine adult bearded dragons (Pogona vitticeps) with cholelithiasis between 2013 and 2015 in southern Germany.
With an increased number of pet reptiles, many diseases occur due to nutritional disorders. Between 2013 and 2015, irregular gallbladder contents (sludge/choleliths) in adult bearded dragons (Pogona vitticeps) were recorded in many of the routinely conducted necropsies at the reptile rescue station in Munich (Auffangstation für Reptilien, München e.V., Munich, Germany). Nine animals, six from the rescue station and three from an associated veterinary practice (Tierärztliche Praxis für Exoten, Augsburg, Germany), were studied. Gallbladder contents from all animals were analysed at the Institute for Clinical Chemistry in Zurich, Switzerland. In three of nine animals, one cholelith composed of 100% calcium carbonate (CaCO3 ) was detected and it precipitated either as pure calcite or as a calcite:vaterite combination. In the remaining six animals, analyses suggested a protein-based material. The detection of choleliths/sludge was not anticipated at necropsy or surgery in eight of nine animals. The diet of the six animals from the rescue station was retrospectively described as mainly insects, whereas the diet of the three animals from the veterinary practice also contained little plant matter. Fed insect species were mealworm larva (Tenebrio molitor), house cricket (Acheta domestica), migratory locust (Locusta migratoria) and zophobas larva (Zophobas morio), all high in protein and fat. In other species, a nidus must be present for CaCO3 to precipitate. As a protein-based sludge was detected in six gallbladders, it is possible that a high-protein diet could lead to such a nidus and subsequently to cholelith formation. Cholelithiasis seems to be a rising problem in adult bearded dragons and is likely underdiagnosed, as many choleliths were found at necropsy. This rise in cholelithiasis may correlate with an unnatural high-protein, high-fat insect-based diet instead of a balanced plant-based diet.


Ecological and geographical overlap drive plumage evolution and mimicry in woodpeckers.
Organismal appearances are shaped by selection from both biotic and abiotic drivers. For example, Gloger's rule describes the pervasive pattern that more pigmented populations are found in more humid areas. However, species may also converge on nearly identical colours and patterns in sympatry, often to avoid predation by mimicking noxious species. Here we leverage a massive global citizen-science database to determine how biotic and abiotic factors act in concert to shape plumage in the world's 230 species of woodpeckers. We find that habitat and climate profoundly influence woodpecker plumage, and we recover support for the generality of Gloger's rule. However, many species exhibit remarkable convergence explained neither by these factors nor by shared ancestry. Instead, this convergence is associated with geographic overlap between species, suggesting occasional strong selection for interspecific mimicry.


[Re-implantation of the fallopian tubes].
Surgical re-implantation of the Fallopian tubes into the uterine cavity has almost become redundant in view of the improved technique of microsurgical tuboplasty. A series of 10 cases are reported in which the conventional method of re-implantation was employed. The successful pregnancy rate of 60% compares favourably with the best results obtained by microsurgery.


Antitumor, immunomodulatory and anti-HIV effect of mangiferin, a naturally occurring glucosylxanthone.
Mangiferin, a C-glucosylxanthone (1,3,6,7-tetrahydroxyxanthone-C2-beta-D-glucoside) purified from plant sources was shown to have in vivo growth-inhibitory activity against ascitic fibrosarcoma in Swiss mice. Following in vivo or in vitro treatment, it also enhanced tumor cell cytotoxicity of the splenic cells and peritoneal macrophages of normal and tumor-bearing mice. In vitro treatment of the splenic cells of tumor-bearing mice with mangiferin resulted in augmented killing of tumor cells, both resistant and sensitive to natural killer cells. Mangiferin was also found to antagonize in vitro the cytopathic effect of HIV. The drug appears to act as a potent biological response modifier with antitumor and antiviral effect.


Trucking organization and mental health disorders of truck drivers.
There are over 3 million truck drivers employed in the commercial transportation and material moving occupations, one of the largest occupational groups in the United States. Workers in this large and growing occupational segment are at risk for a range of occupational health-induced conditions, including mental health and psychiatric disorders due to high occupational stress, low access and use of health care, and limited social support. The purpose of this study was to explore male truck drivers' mental health risks and associated comorbidities, using a cross-sectional and quantitative design. Data were collected from a random sample of 316 male truckers between the ages of 23 and 76 at a large truck stop located within a 100-mile radius of Greensboro, North Carolina, USA, using a self-administered 82-item questionnaire. Surveyed truckers were found to have significant issues affecting their mental health, such as loneliness (27.9%), depression (26.9%), chronic sleep disturbances (20.6%), anxiety (14.5%), and other emotional problems (13%). Findings have potential to help researchers develop interventions to improve the emotional and occupational health of truck drivers, a highly underserved population. Mental health promotion, assessment, and treatment must become a priority to improve the overall trucking environment for truckers, the transportation industry, and safety on US highways.


Biocompatible Silk/Polymer Energy Harvesters Using Stretched Poly (vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) Nanofibers.
Energy harvested from human body movement can produce continuous, stable energy to portable electronics and implanted medical devices. The energy harvesters need to be light, small, inexpensive, and highly portable. Here we report a novel biocompatible device made of poly (vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) nanofibers on flexible substrates. The nanofibers are prepared with electrospinning followed by a stretching process. This results in aligned nanofibers with diameter control. The assembled device demonstrates high mechanical-to-electrical conversion performance, with stretched PVDF-HFP nanofibers outperforming regular electrospun samples by more than 10 times. Fourier transform infrared spectroscopy (FTIR) reveals that the stretched nanofibers have a higher β phase content, which is the critical polymorph that enables piezoelectricity in polyvinylidene fluoride (PVDF). Polydimethylsiloxane (PDMS) is initially selected as the substrate material for its low cost, high flexibility, and rapid prototyping capability. Bombyx Mori silkworm silk fibroin (SF) and its composites are investigated as promising alternatives due to their high strength, toughness, and biocompatibility. A composite of silk with 20% glycerol demonstrates higher strength and larger ultimate strain than PDMS. With the integration of stretched electrospun PVDF-HFP nanofibers and flexible substrates, this pilot study shows a new pathway for the fabrication of biocompatible, skin-mountable energy devices.


IL-7 signaling imparts polyfunctionality and stemness potential to CD4(+) T cells.
The functional status of CD4(+) T cells is a critical determinant of antitumor immunity. Polyfunctional CD4(+) T cells possess the ability to concomitantly produce multiple Th1-type cytokines, exhibiting a functional attribute desirable for cancer immunotherapy. However, the mechanisms by which these cells are induced are neither defined nor it is clear if these cells can be used therapeutically to treat cancer. Here, we report that CD4(+) T cells exposed to exogenous IL-7 during antigenic stimulation can acquire a polyfunctional phenotype, characterized by their ability to simultaneously express IFNγ, IL-2, TNFα and granzyme B. This IL-7-driven polyfunctional phenotype was associated with increased histone acetylation in the promoters of the effector genes, indicative of increased chromatin accessibility. Moreover, forced expression of a constitutively active (CA) form of STAT5 recapitulated IL-7 in inducing CD4(+) T-cell polyfunctionality. Conversely, the expression of a dominant negative (DN) form of STAT5 abolished the ability of IL-7 to induce polyfunctional CD4(+) T cells. These in-vitro-generated polyfunctional CD4(+) T cells can traffic to tumor and expand intratumorally in response to immunization. Importantly, adoptive transfer of polyfunctional CD4(+) T cells following lymphodepletive chemotherapy was able to eradicate large established tumors. This beneficial outcome was associated with the occurrence of antigen epitope spreading, activation of the endogenous CD8(+) T cells and persistence of donor CD4(+) T cells exhibiting memory stem cell attributes. These findings indicate that IL-7 signaling can impart polyfunctionality and stemness potential to CD4(+) T cells, revealing a previously unknown property of IL-7 that can be exploited in adoptive T-cell immunotherapy.


Resilience in the face of fragile X syndrome.
In this article, we communicate transformative findings from a case study on the resilience of a young woman with fragile X syndrome (FXS), a genetic condition involving mental impairment and physical, emotional, and behavioral challenges. We explored the resilience of "Lucy," a spirited 16-year-old North American, using informal interviews with her, formal interviews with significant adults in her life, and observations (visual and anecdotal) over 20 months. In reporting the information-rich case of Lucy, well supported by her ecology to rise above full-mutation FXS, we encourage a positive perspective of living with FXS. Although we recognize the limitations of a single case study, our findings offer tentative, process-oriented insights into resilience in contexts of genetic disability, previously unreported in conjunction with FXS. We concluded that the processes informing Lucy's resilience were partly her responsibility and partly her social ecology's, and comprised intrapersonal agency, unconditional positive acceptance and belonging, and support toward mastery.


Impact of Overlying Personal Items on CT Dose with Use of Automated Tube Current Modulation-Pilot Investigation.
To determine the incidence and impact of overlying radiopaque personal items (e.g., cellular phones, zippers) on CT dose and image quality with use of automated tube current modulation. Topogram images from 100 consecutive adult outpatient CT abdomen pelvis studies were retrospectively reviewed, and the number and type of overlying radiopaque personal items were recorded. Additionally, an anthropomorphic phantom was imaged with overlying personal items 1) present in topogram and axial images; 2) present in topogram but removed prior to axial acquisition; and 3) present in topogram positioned outside the field of view of the axial acquisition. dose length product (DLP) and CT dose index volume (CTDIvol) were compared to acquisitions performed without overlying personal items. Image noise was evaluated by assessing the standard deviation of Hounsfield units at the level of the overlying personal item. Overlying personal items were visible in topogram images for 55% of CT exams and included underwires (38% of exams), zippers (7%), and cellular phones (1%). DLP increased when a cellular phone was present in the topogram whether or not it was removed before axial image acquisition (3.7% p = 0.002, combined AutomA and SmartmA), and image noise increased (144%, p = 0.002; AutomA). No increase in dose or image noise was observed with overlying zippers or underwires or when any object was visible in the topogram outside the field of view of the axial images. Overlying personal items were observed in the majority of abdominopelvic CT scans. Large overlying radiopaque personal items resulted in increased dose and increased image noise. Removal of all overlying personal items will result in optimized dose and image quality.


Hypertension, cardiac hypertrophy, and sudden death in mice lacking natriuretic peptide receptor A.
Natriuretic peptides, produced in the heart, bind to the natriuretic peptide receptor A (NPRA) and cause vasodilation and natriuresis important in the regulation of blood pressure. We here report that mice lacking a functional Npr1 gene coding for NPRA have elevated blood pressures and hearts exhibiting marked hypertrophy with interstitial fibrosis resembling that seen in human hypertensive heart disease. Echocardiographic evaluation of the mice demonstrated a compensated state of systemic hypertension in which cardiac hypertrophy and dilatation are evident but with no reduction in ventricular performance. Nevertheless, sudden death, with morphologic evidence indicative in some animals of congestive heart failure and in others of aortic dissection, occurred in all 15 male mice lacking Npr1 before 6 months of age, and in one of 16 females in our study. Thus complete absence of NPRA causes hypertension in mice and leads to cardiac hypertrophy and, particularly in males, lethal vascular events similar to those seen in untreated human hypertensive patients.


Premature delivery influences the immunological composition of colostrum and transitional and mature human milk.
Human breast milk is the ideal nutrition for the newborn, and in addition to its nutritional contribution, necessary for infant growth and development, it contains various immune bioactive factors that confer some of the numerous beneficial effects of breastfeeding. The current study analyzed the concentrations of IgA, growth factors such as epidermal growth factor (EGF), TGFβ1, and TGFβ2, cytokines IL-6, IL-8, IL-10, IL-13, and TNFα, and TNF-receptor I (TNF-RI) in colostrum and transitional and mature milk from mothers with mature, premature, and very premature infants. Human milk samples were collected from mothers delivering at term (T), preterm (PT), and very preterm (VPT). Milk from all the mothers was collected at 3 different time points after delivery corresponding to colostrum and transitional and mature milk. After obtaining milk whey, IgA, EGF, TGFβ1, and TGFβ2 were determined by ELISA and IL-6, IL-8, IL-10, IL-13, TNFα and TNF-RI by cytometric bead array immunoassay. The colostrum of the PT group was extremely rich in most of the factors studied, but higher concentrations than in the T group were only found for IL-6 (P = 0.051), TGFβ1, and TGFβ2 (P < 0.05). Conversely, the colostrum of the VPT group had lower concentrations of IgA, IL-8, IL-10, and TNFα than those in the T group (P < 0.05). Results suggest that maternal lactogenic compensatory mechanisms accelerating the development of immature breast-fed preterm infants may take effect only after wk 30 of gestation.


Multicenter urethroplasty outcomes for urethral stricture disease for patients with neurogenic bladder or bladder dysfunction requiring clean intermittent catheterization.
Our objective is to better comprehend treatment considerations for urethral stricture disease (USD) in patients requiring long-term clean intermittent catheterization (CIC). Patient characteristics, surgical outcomes and complications are unknown in this population. Six members of the Trauma and Urologic Reconstruction Network of Surgeons (TURNS) participated in a prospective (2009 to present) and retrospective (prior to 2009) database recording patient demographics, surgical approach and outcomes. We included all patients undergoing urethroplasty who perform CIC. Descriptive statistics were used to analyze results. A total of 37 patients with 39 strictures were included. Bladder dysfunction was characterized as detrusor failure in 35% and neurogenic etiology in 65%. Median stricture length was 3 cm (IQR: 1.5-5.5) with 28% repaired with dorsal onlay buccal mucosal graft, 26% excision and primary anastomosis, 8% dorsal inlay, 8% ventral and dorsal, 8% flap based 8% non-transecting and 15% other. Functional success was 90%: 4 patients required DVIU or dilation due to recurrence, with 2 of those ultimately requiring repeat urethroplasty. 86% of patients returned to CIC; no patients reported new pad use for urinary leakage after urethroplasty. During a median follow-up period of 3.1 years (IQR: 1.0-5.3), no patients underwent urinary diversion. Urethroplasty is suitable, safe and effective for patients dependent on CIC suffering from USD. The effect of continual CIC on long-term outcomes remains uncertain.


Acute Upper-Limb Complications Following Radial Artery Catheterization for Coronary Angiography.
The radial artery is becoming the vessel of choice for performing cardiac catheterization. Transradial catheterization can impose risks on the upper extremity, and hand surgeons should be aware of the most frequent complications. The purpose of this study was to determine the frequency, timing, and scope of upper-limb complications shortly after transradial catheterization. A retrospective review was conducted of the medical records of patients who underwent catheterization between 2009 and 2016. Complications were assessed for up to 60 days. The Cox model was used to assess risk factors for complications. A total of 10,540 patients were included in the analysis (68.5% male), median age 67 years. There were 79 patients who experienced at least one complication within 60 days (0.84% of procedures; 95% confidence interval, 0.65% to 1.02%). The most common complications were hematoma (n = 39) and radial artery occlusion (n = 28). Other complications included pseudoaneurysm (n = 7), arteriovenous fistula (n = 3), carpal tunnel syndrome (n = 4), arterial perforation (n = 3), persistent vasospasm (n = 2), and compartment syndrome (n = 1). The complications were diagnosed a median of 1 day after catheterization. Female sex was at increased risk for developing a complication. Diabetes, age, body mass index, and catheter size were not associated with an increased risk for developing a complication. Ten patients underwent surgical management of a complication. Reasons for surgery included symptomatic radial artery occlusions, pseudoaneurysm formation, arteriovenous fistulas, and compartment syndrome. No identifiable risk factors were associated with patients who underwent surgical intervention. The frequency of upper-limb complications after radial artery catheterization is small. They include arterial occlusion, bleeding, compartment syndrome, arteriovenous fistula, and pseudoaneurysm. Most complications presented within 1 week of the procedure and occurred more frequently in the female sex. Operative management of complications was infrequent. Therapeutic IV.


Detection of membrane cholesterol by filipin in isolated rat liver coated vesicles is dependent upon removal of the clathrin coat.
We investigated the cholesterol content of highly purified populations of coated vesicles from rat liver by biochemical quantitation and by cytochemical electron microscopy using the polyene antibiotic filipin. Failure of this reagent to elicit its typical response for a cholesterol-containing membrane, i.e., a characteristically corrugated or rippled appearance by thin section analysis, had led to the hypothesis (Montesano, R., A. Perrelet, P. Vassalli, and L. Orci, 1979, Proc. Natl. Acad. Sci. USA., 76:6391-6395) that cholesterol is specifically excluded from the plasma membrane domains associated with coated pit regions. The present electron microscopic results showed that although the response of coated vesicle membranes to filipin was also negative, uncoated vesicles whose clathrin coats had been removed in vitro exhibited a strong filipin-positive response. Quantitated biochemically, the cholesterol-to-phospholipid ratio of the coated vesicles was found to be indistinguishable from that of control preparations of plasma membranes isolated from rat liver. Taken together, the results indicate that the filipin-negative response of coated vesicles (and probably also that of coated pits) is due not to abnormally low cholesterol content, but rather to the stabilizing influence of their enveloping clathrin coats which inhibit the characteristic structural expression of the filipin-cholesterol complexes.


Paclitaxel-loaded chitosan oligosaccharide-stabilized gold nanoparticles as novel agents for drug delivery and photoacoustic imaging of cancer cells.
Polymer nanoparticles have gained significant attention as potential drug carriers for anticancer agents and molecular imaging. Biocompatible gold nanoparticles (AuNPs) were synthesized using chitosan oligosaccharide (COS) as a reducing and stabilizing agent and were subsequently loaded with paclitaxel (PTX) to demonstrate their use in drug delivery and photoacoustic imaging (PAI) of MDA-MB-231 cells. Paclitaxel-loaded chitosan oligosaccharide-stabilized gold nanoparticles (PTX-COS AuNPs) were spherical in shape with an average particle size of 61.86±3.01nm. PTX-COS AuNPs showed sustained and pH-dependent drug release profiles and exhibited strong cytotoxic effect against MDA-MB-231 cells through the induction of apoptosis with improved reactive oxygen species (ROS) generation and altered mitochondrial membrane potential (MMP) level. The cellular internalization of PTX-COS AuNPs was proven by fluorescence microscopy as well as flow cytometry. PTX-COS AuNPs were also evaluated as a new class of optical contrast agents for photoacoustic imaging (PAI). To the best of our knowledge, this is the first report that describes the use of PTX-COS AuNPs as novel agents for drug delivery and PAI of cancer cells. These results exposed the promising potential of PTX-COS AuNPs in the field of drug delivery, molecular imaging, and cancer therapy in the near future.


Volatile organic compounds in the unsaturated zone from radioactive wastes.
Volatile organic compounds (VOCs) are often comingled with low-level radioactive wastes (LLRW), but little is known about subsurface VOC emanations from LLRW landfills. The current study systematically quantified VOCs associated with LLRW over an 11-yr period at the USGS Amargosa Desert Research Site (ADRS) in southwestern Nevada. Unsaturated-zone gas samples of VOCs were collected by adsorption on resin cartridges and analyzed by thermal desorption and GC/MS. Sixty of 87 VOC method analytes were detected in the 110-m-thick unsaturated zone surrounding a LLRW disposal facility. Chlorofluorocarbons (CFCs) were detected in 100% of samples collected. Chlorofluorocarbons are powerful greenhouse gases, deplete stratospheric ozone, and are likely released from LLRW facilities worldwide. Soil-gas samples collected from a depth of 24 m and a horizontal distance 100 m south of the nearest waste-disposal trench contained >60,000 ppbv total VOCs, including >37,000 ppbv CFCs. Extensive sampling in the shallow unsaturated zone (0-2 m deep) identified areas where total VOC concentrations exceeded 5000 ppbv at the 1.5-m depth. Volatile organic compound concentrations exceeded background levels up to 300 m from the facility. Maximum vertical diffusive fluxes of total VOCs were estimated to be 1 g m yr. Volatile organic compound distributions were similar but not identical to those previously determined for tritium and elemental mercury. To our knowledge, this study is the first to characterize the unsaturated zone distribution of VOCs emanating from a LLRW landfill. Our results may help explain anomalous transport of radionuclides at the ADRS and elsewhere.


Effects of iron and carbon monoxide on Lachesis muta muta venom-mediated degradation of plasmatic coagulation.
Hypofibrinogenemia is an important clinical consequence following envenomation by Lachesis muta muta, usually attenuated or prevented by administration of antivenom. The venom of L. m. muta contains both a metalloproteinase fibrinogenase and a serine protease thrombin-like enzyme, and exposure of fibrinogen to iron (Fe) and carbon monoxide (CO) has been demonstrated to decrease its catalysis by such enzymes. Using thrombelastographic analytical techniques, it was determined that this venom displayed weak procoagulant effects combined with fibrinogenolytic effects, and pretreatment of plasma with Fe and CO markedly attenuated venom-mediated effects. Additional experiments involving heparin exposure and varying calcium concentrations demonstrated that modification of fibrinogen with Fe and CO in human plasma rendered fibrinogen not recognizable to the fibrinogenolytic metalloproteinase but did not prevent polymerization by the thrombin-like serine protease. Lastly, when venom was exposed to CO in isolation and then placed in plasma, the fibrinogenase was inhibited but the thrombin-like enzyme was not inhibited. In sum, utilizing relatively facile modifications, we demonstrated with thrombelastography that Fe and/or CO addition can protect human plasmatic coagulation from fibrinogenase activity but not the effects of the thrombin-like activity of L. m. muta venom.


Regional differences in incidence and patient characteristics of moyamoya disease: a systematic review.
Moyamoya disease (MMD) is a rare cause of stroke, initially described in Japan. In other countries, incidences and presenting symptoms may differ from those in Japan. The literature on regional differences in incidence and patient characteristics of MMD was systematically reviewed. Medline, EMBASE and CINAHL were searched for population based studies on MMD published between January 1969 and January 2011. From studies that met predefined inclusion criteria, information was extracted on incidence and patient characteristics. Incidences with corresponding 95% CIs if possible were calculated and descriptive statistics for patient characteristics were used. 8 studies were included: three from Japan, one each from Taiwan and China and three from the USA. Incidences per 100 000 patient years ranged in Japan from 0.35 to 0.94 (95% CI 0.69 to 1.19), in the USA from 0.05 (-0.04 to 0.12) in Iowa to 0.17 (-0.06 to 0.40) in Hawaii and were 0.41 (0.28 to 0.54) in Nanjing, China and 0.02 (0.003 to 0.04) in Taiwan. Female to male ratio ranged from 1.1 (0.9 to 1.5) in Nanjing to 2.8 (1.2 to 6.1) in Iowa. Proportions with intracerebral haemorrhage as the initial presentation were 56% in China, 52% in Taiwan, 29% in Hawaii, 21% in Japan and 10% in Iowa. Patients with childhood onset presented most often with ischaemia (>75%) in all regions. MMD incidence was higher in Japan and China than in Taiwan and North America and presenting symptoms showed regional differences, which are thus far unexplained. Population based data on MMD in Europe are lacking.


Anosognosia for hemiplegia after stroke is a multifaceted phenomenon: a systematic review of the literature.
Anosognosia is the lack of awareness or the underestimation of a specific deficit in sensory, perceptual, motor, affective or cognitive functioning due to a brain lesion. This self-awareness deficit has been studied mainly in stroke hemiplegic patients, who may report no deficit, overestimate their abilities or deny that they are unable to move a paretic limb. In this review, a detailed search of the literature was conducted to illustrate clinical manifestations, pathogenetic models, diagnostic procedures and unresolved issues in anosognosia for motor impairment after stroke. English and French language papers spanning the period January 1990-January 2007 were selected using PubMed Services and utilizing research words stroke, anosognosia, awareness, denial, unawareness, hemiplegia. Papers reporting sign-based definitions, neurological and neuropsychological data and the results of clinical trials or historical trends in diagnosis were chosen. As a result, a very complex and multifaceted phenomenon emerges, whose variable behavioural manifestations often produce uncertainties in conceptual definitions and diagnostic procedures. Although a number of questionnaires and diagnostic methods have been developed to assess anosognosia following stroke in the last 30 years, they are often limited by insufficient discriminative power or a narrow focus on specific deficits. As a consequence, epidemiological estimates are variable and incidence rates have ranged from 7 to 77% in stroke. In addition, the pathogenesis of anosognosia is widely debated. The most recent neuropsychological models have suggested a defect in the feedforward system, while neuro-anatomical studies have consistently reported on the involvement of the right cerebral hemisphere, particularly the prefrontal and parieto-temporal cortex, as well as insula and thalamus. We highlight the need for a multidimensional assessment procedure and suggest some potentially productive directions for future research about unawareness of illness.


Metagenomic evidence for reciprocal particle exchange between the mainstem estuary and lateral bay sediments of the lower Columbia River.
Lateral bays of the lower Columbia River estuary are areas of enhanced water retention that influence net ecosystem metabolism through activities of their diverse microbial communities. Metagenomic characterization of sediment microbiota from three disparate sites in two brackish lateral bays (Baker and Youngs) produced ∼100 Gbp of DNA sequence data analyzed subsequently for predicted SSU rRNA and peptide-coding genes. The metagenomes were dominated by Bacteria. A large component of Eukaryota was present in Youngs Bay samples, i.e., the inner bay sediment was enriched with the invasive New Zealand mudsnail, Potamopyrgus antipodarum, known for high ammonia production. The metagenome was also highly enriched with an archaeal ammonia oxidizer closely related to Nitrosoarchaeum limnia. Combined analysis of sequences and continuous, high-resolution time series of biogeochemical data from fixed and mobile platforms revealed the importance of large-scale reciprocal particle exchanges between the mainstem estuarine water column and lateral bay sediments. Deposition of marine diatom particles in sediments near Youngs Bay mouth was associated with a dramatic enrichment of Bacteroidetes (58% of total Bacteria) and corresponding genes involved in phytoplankton polysaccharide degradation. The Baker Bay sediment metagenome contained abundant Archaea, including diverse methanogens, as well as functional genes for methylotrophy and taxonomic markers for syntrophic bacteria, suggesting that active methane cycling occurs at this location. Our previous work showed enrichments of similar anaerobic taxa in particulate matter of the mainstem estuarine water column. In total, our results identify the lateral bays as both sources and sinks of biogenic particles significantly impacting microbial community composition and biogeochemical activities in the estuary.


Importance of a martian hematite site for astrobiology.
Defining locations where conditions may have been favorable for life is a key objective for the exploration of Mars. Of prime importance are sites where conditions may have been favorable for the preservation of evidence of prebiotic or biotic processes. Areas displaying significant concentrations of the mineral hematite (alpha-Fe2O3), recently identified by thermal emission spectrometry, may have significance in the search for evidence of extraterrestrial life. Since iron oxides can form as aqueous mineral precipitates, the potential exists to preserve microscopic evidence of life in iron oxide-depositing ecosystems. Terrestrial hematite deposits proposed as possible analogs for hematite deposits on Mars include massive (banded) iron formations, iron oxide hydrothermal deposits, iron-rich laterites and ferricrete soils, and rock varnish. We report the potential for long-term preservation of microfossils by iron oxide mineralization in specimens of the approximately 2,100-Ma banded iron deposit of the Gunflint Formation, Canada. Scanning and analytical electron microscopy reveals micrometer-scale rods, spheres, and filaments consisting predominantly of iron and oxygen with minor carbon. We interpret these objects as microbial cells permineralized by an iron oxide, presumably hematite. The confirmation of ancient martian microbial life in hematite deposits will require the return of samples to terrestrial laboratories. A hematite-rich deposit composed of aqueous iron oxide precipitates may thus prove to be a prime site for future sample return.


[Traumatic atlanto-occipital dislocation in a calf].
In a 7-week-old calf neurologic symptoms occurred after an accident in the feeding grid. The calf was alert but in left lateral recumbency. After having been placed on its right side it showed a sideward drift to the left. Following head movement, an asynchronous movement of the eyes could be observed. Lesions of the upper motor neuron system, N. vestibulocochlearis, Nn. oculomotorius, trochlearis and/or abducens as well as N. vagus were suspected. Radiological examination of the occipital region and cervical spine revealed an atlanto-occipital subluxation of the atlas. Based on all these findings, the diagnosis of traumatic atlanto-occipital dislocation was made and the calf was euthanized. The pathological findings confirmed this diagnosis.


Massive upper gastrointestinal bleeding secondary to duodenal metastasis of transitional cell carcinoma of the urinary bladder.
Acute upper gastrointestinal (UGI) bleeding is a common problem in our clinical practice and is often due to peptic ulcer diseases. Occasionally, malignancy may be implicated in these situations. Here we report a rare case of UGI bleeding secondary to metastatic transitional cell carcinoma (TCC) of the urinary bladder. A 62-year-old man with a history of stage IIIb TCC of the urinary bladder presented with hematemesis. Endoscopy showed a large tumor in the second stage of the duodenum that occupied 40% of the duodenal circumference, over 7 cm in length. Biopsies revealed a poorly differentiated malignant neoplasm consistent with metastasis from urothelial carcinoma that was identical to the previous surgical specimen of the urinary bladder. He was treated with supportive therapy and intravenous proton pump inhibitor and was discharged home 2 weeks later. Two weeks after discharge, the patient returned to the hospital with a painful swelling of the floor of his mouth. Biopsy again showed the same cancer type. He had unremitting bleeding from his mouth requiring multiple transfusions and a course of palliative radiation therapy. He progressively deteriorated in his cardiopulmonary and neurological functions and expired with cardiopulmonary arrest one month later.


Canadian Update on Surgical Procedures (CUSP) Urology Group consensus for intraoperative hemostasis during minimally invasive partial nephrectomy.
Partial nephrectomy remains the gold standard in the management of small renal masses. However, minimally invasive partial nephrectomy (MIPN) is associated with a steep learning curve, and optimal, standardized techniques for time-efficient hemostasis are poorly described. Given the relative lack of evidence, the goal was to describe a set of actionable guiding principles, through an expert working panel, for urologists to approach hemostasis without compromising warm ischemia or oncological outcomes. A three-step modified Delphi method was used to achieve expert agreement on the best practices for hemostasis in MIPN. Panelists were recruited from the Canadian Update on Surgical Procedures (CUSP) Urology Group, which represent all provinces, academic and community practices, and fellowship-and non-fellowship-trained surgeons. Thirty-two (round 1) and 46 (round 2) panellists participated in survey questionnaires, and 22 attended the in-person consensus meeting. An initial literature search of 945 articles (230 abstracts) underwent screening and yielded 24 preliminary techniques. Through sequential survey assessment and in-person discussion, a total of 11 strategies were approved. These are temporally distributed prior to tumor resection (five principles), during tumor resection (two principles), and during renorrhaphy (four principles). Given the variability in tumor size, depth, location, and vascularity, coupled with limitations of laparoscopic equipment, achieving consistent hemostasis in MIPN may be challenging. Despite over two decades of MIPN experience, limited evidence exists to guide clinicians. Through a three-step Delphi method and rigorous iterative review with a panel of experts, we ascertained a guiding checklist of principles for newly beginning and practicing urologists to reference.


An unusual cause of fungal pneumonia.
A 65 year old female, known asthmatic on steroids intermittently, with no other co-morbidity presented with fever, breathlessness and cough with mucoid expectoration of ten days duration with bilateral crepts, went for Type II respiratory failure and was intubated followed by tracheostomy in view of prolonged ventilator support. In spite of high end antibiotics as per sputum culture sensitivity, weaning off the ventilator was not possible. Blood investigations revealed leucocytosis with neutrophilic predominance and IgE levels were within normal limits. CT chest showed multiple patchy consolidations of the right upper, middle and lower lobes with ground glass appearance and enlarged mediastinal lymph nodes. Work up for retrovirus, tuberculosis and Sputum for KOH mount was negative. No evidence of sputum and blood eosinophilia. BAL sample grew Curvularia species. Fluconazole 150mg OD was added. Serial imaging of the chest showed resolution of the consolidation and was weaned off the ventilator and was comfortable on room air. Pneumonia caused by Curvularia, in an immune competent patient is very rare. Even in broncho pulmonary involvement these fungi usually occur in allergic conditions as in ABPA than appearing as a solitary cause for lung infection. But if diagnosed and treated early, will respond well to triazoles. This case report highlights a unilateral fungal pneumonia with dramatic clinical improvement post treatment once the rare causative organism was identified.