The terminal nerve plays a prominent role in GnRH-1 neuronal migration independent from proper olfactory and vomeronasal connections to the olfactory bulbs.
Gonadotropin-releasing hormone-1 (GnRH-1) neurons (GnRH-1 ns) migrate from the developing olfactory pit into the hypothalamus during embryonic development. Migration of the GnRH-1 neurons is required for mammalian reproduction as these cells control release of gonadotropins from the anterior pituitary gland. Disturbances in GnRH-1 ns migration, GnRH-1 synthesis, secretion or signaling lead to varying degrees of hypogonadotropic hypogonadism (HH), which impairs pubertal onset and fertility. HH associated with congenital olfactory defects is clinically defined as Kallmann Syndrome (KS). The association of olfactory defects with HH in KS suggested a potential direct relationship between defective olfactory axonal routing, lack of olfactory bulbs (OBs) and aberrant GnRH-1 ns migration. However, it has never been experimentally proven that the formation of axonal connections of the olfactory/vomeronasal neurons to their functional targets are necessary for the migration of GnRH-1 ns to the hypothalamus. Loss-of-function of the Arx-1 homeobox gene leads to the lack of proper formation of the OBs with abnormal axonal termination of olfactory sensory neurons ( Yoshihara et al., 2005). Our data prove that correct development of the OBs and axonal connection of the olfactory/vomeronasal sensory neurons to the forebrain are not required for GnRH-1 ns migration, and suggest that the terminal nerve, which forms the GnRH-1 migratory scaffold, follows different guidance cues and differs in gene expression from olfactory/vomeronasal sensory neurons.


Safety and Pharmacokinetics of High-Dose TAS-303 in Healthy Japanese Volunteers: A Single-Center, Single-Blind, Randomized, Placebo-Controlled, Parallel-Group, Multiple-Ascending-Dose Study.
Preclinical data of TAS-303 (4-piperidinyl 2,2-diphenyl-2-[propoxy-1,1,2,2,3,3,3-d7 ] acetate hydrochloride), a noradrenaline reuptake inhibitor, show that it increases urethral contraction in rats and may therefore benefit stress urinary incontinence patients. In this single-blind, randomized, placebo-controlled, parallel-group, multiple-ascending-dose phase 1 study, we evaluated the safety and tolerability of once-daily TAS-303 8, 10, 12, 15, or 18 mg administered for 16 days in healthy subjects. In addition, we investigated the pharmacokinetics and inhibitory effect of TAS-303 on hepatic cytochrome P450 (CYP) 3A activity. Rates of adverse events, adverse drug reactions, and pharmacokinetic parameters of TAS-303 were evaluated. Fifty subjects were randomized: 7 subjects each were assigned to receive TAS-303 8-18 mg, and 3 subjects each were assigned to receive placebo at each dose. The overall incidences of adverse events and adverse drug reactions in all subjects administered TAS-303 (n = 35) was 25.7% and 2.9%, respectively, and those for the placebo groups (n = 15) were 46.7% and 0%, respectively. No deaths or serious adverse events occurred. TAS-303 displayed a dose-proportional pharmacokinetic profile across doses of 8-18 mg over the 16-day multiple administration period, and TAS-303 might inhibit hepatic CYP3A activity within this dose range. TAS-303 at a dose of 8-18 mg was confirmed to be safe and tolerable.


Oyster (Crassostrea gigas) ferritin should be a promising Fe2+ nanocarrier.
Ferritin, one of the storage forms of iron, which widely exists in all living organisms and shows its potential as functional nanomaterial. In this study, Fe2+ were added to oyster ferritin (GF1) in vitro until reached experimental saturation state. The structure characterization and monodisperse morphology of ferritin were showed that the protein structure of ferritin did not change significantly after adding Fe2+ and GF1 with 1000 Fe2+ (GF1-1000 Fe2+) was selected for subsequent experiments. The storage stability results showed that 11.27 % of the protein remained cage-like structure when GF1-1000 Fe2+ was stored at 4 °C for 12 days. The thermal treatment results showed that GF1-1000 Fe2+ still maintained certain structure and activity at 80 °C, and 82.8 % of Fe2+ was still retained at 80 °C. These experimental results may provide a theoretical basis for the development of novel iron supplement that are non-toxic and highly stable.


Algal origin of sponge sterane biomarkers negates the oldest evidence for animals in the rock record.
The earliest fossils of animal-like organisms occur in Ediacaran rocks that are approximately 571 million years old. Yet 24-isopropylcholestanes and other C30 fossil sterol molecules have been suggested to reflect an important ecological role of demosponges as the first abundant animals by the end of the Cryogenian period (>635 million years ago). Here, we demonstrate that C30 24-isopropylcholestane is not diagnostic for sponges and probably formed in Neoproterozoic sediments through the geological methylation of C29 sterols of chlorophyte algae, the dominant eukaryotes at that time. These findings reconcile biomarker evidence with the geological record and revert the oldest evidence for animals back into the latest Ediacaran.


High coagulation factor VIII and von Willebrand factor in patients with lymphoma and leukemia.
The risk of venous thromboembolism is increased in patients with lymphoma and leukemia; however, little is known about the potential underlying hereditary or acquired thrombophilia. We prospectively analyzed procoagulant markers and gene mutations in patients with lymphoma (n = 35) and leukemia (n = 10) at diagnosis and over the course of treatment. Global coagulation tests were normal in all patients, as were antithrombin and protein S. Activated protein C resistance caused by the factor V Leiden mutation was found in four patients, one patient had the G20210A mutation of the prothrombin gene, and one patient had protein C deficiency. The most striking findings were sustained very high levels of factor VIII (>150 %) in 30 patients (68 %), which correlated with high von Willebrand factor. An acute phase response in these patients was ruled out by absence of fever and normal IL-6 and -α. Elevated factor VIII is an independent thrombophilic risk factor and may play an etiologic role in thromboembolic complications in patients with malignant lymphoma. Since high von Willebrand factor is most likely caused by endothelial cell injury, an additional, unknown pathophysiological association with malignant lymphoma and acute leukemia is possible.


Deadly crosstalk: Notch signaling at the intersection of EMT and cancer stem cells.
Notch signaling is an evolutionarily conserved pathway involved in cell fate control during development, stem cell self-renewal and postnatal tissue differentiation. Roles for Notch in carcinogenesis, in the biology of cancer stem cells, tumor angiogenesis and epithelial-to-mesenchymal transition (EMT) have been reported. This mini-review describes the role of Notch signaling deregulation in EMT and tumor aggressiveness. We describe how accumulated evidence suggests that Notch inhibition is an attractive strategy for the treatment of several cancers, at least in part because of its potential to reverse or prevent EMT.


Unusual subdural hematoma after SPA use.
We report a case of right tentorium subdural hematoma (SDH) in a 72-year-old man a few days after he visited a sanitas per aqua (SPA) facility. He had directed high-pressure water onto his head and neck. The patient was admitted to the emergency room during his 3rd day of severe throbbing and generalized headache. Cerebral computed tomography revealed a high-density SDH over the right tentorium. This is the first reported case of SPA-related SDH. Neurologists might consider the possibility that SPA use may be a factor in SDH or unexplained neurologic events in otherwise healthy persons.


Lowly expressed lncRNA PVT1 suppresses proliferation and advances apoptosis of glioma cells through up-regulating microRNA-128-1-5p and inhibiting PTBP1.
Glioma is a primary intracranial malignancy with poor prognosis, of which the pathogenesis remains to be elucidated. Therein, the aim of this study is to discuss the impacts of lncRNA plasmacytoma variant translocation 1 (PVT1)/microRNA-128-1-5p (miR-128-1-5p)/polypyrimidine tract-binding protein 1 (PTBP1) axis on the biological characteristics of glioma cells. Glioma tissue samples (72 cases) and normal brain tissue samples (35 cases) were harvested. The expression of PVT1, miR-128-1-5p and PTBP1 in glioma tissues and cells was detected. Glioma cells were transfected with sh-PVT1, miR-128-1-5p mimics or miR-128-1-5p inhibitors to verify the impacts of PVT1 and miR-128-1-5p on DNA damage, cell colony formation, invasion, proliferation, migration and apoptosis of glioma U87 and U251 cells. The growth of transplanted tumor was tested by tumor xenograft in nude mice. The combination of PVT1 and miR-128-1-5p and the targeting relationship between miR-128-1-5p and PTBP1 were verified. PVT1 and PTBP1 expression was enhanced and miR-128-1-5p expression was degraded in glioma tissues and cells. Overexpressed miR-128-1-5p and lowly-expressed PVT1 promoted DNA damage, suppressed colony formation, invasion, proliferation and migration as well as boosted apoptosis of U251 and U87 cells. Up-regulating miR-128-1-5p and down-regulating PVT1 reduced transplanted tumor volume and weight of glioma in mice. Low expression miR-128-1-5p reversed the effect of low expression PVT1 on the biological characteristics of glioma cells. PVT1 specifically bound to miR-128-1-5p and PTBP1 was the target gene of miR-128-1-5p. This study suggests that down-regulated PVT1 or up-regulated miR-128-1-5p boosts apoptosis and attenuates proliferation of glioma cells by inhibiting PTBP1 expression. This study is essential for finding new therapeutic targets for glioma.


Deposition of 1.88-billion-year-old iron formations as a consequence of rapid crustal growth.
Iron formations are chemical sedimentary rocks comprising layers of iron-rich and silica-rich minerals whose deposition requires anoxic and iron-rich (ferruginous) sea water. Their demise after the rise in atmospheric oxygen by 2.32 billion years (Gyr) ago has been attributed to the removal of dissolved iron through progressive oxidation or sulphidation of the deep ocean. Therefore, a sudden return of voluminous iron formations nearly 500 million years later poses an apparent conundrum. Most late Palaeoproterozoic iron formations are about 1.88 Gyr old and occur in the Superior region of North America. Major iron formations are also preserved in Australia, but these were apparently deposited after the transition to a sulphidic ocean at 1.84 Gyr ago that should have terminated iron formation deposition, implying that they reflect local marine conditions. Here we date zircons in tuff layers to show that iron formations in the Frere Formation of Western Australia are about 1.88 Gyr old, indicating that the deposition of iron formations from two disparate cratons was coeval and probably reflects global ocean chemistry. The sudden reappearance of major iron formations at 1.88 Gyr ago--contemporaneous with peaks in global mafic-ultramafic magmatism, juvenile continental and oceanic crust formation, mantle depletion and volcanogenic massive sulphide formation--suggests deposition of iron formations as a consequence of major mantle activity and rapid crustal growth. Our findings support the idea that enhanced submarine volcanism and hydrothermal activity linked to a peak in mantle melting released large volumes of ferrous iron and other reductants that overwhelmed the sulphate and oxygen reservoirs of the ocean, decoupling atmospheric and seawater redox states, and causing the return of widespread ferruginous conditions. Iron formations formed on clastic-starved coastal shelves where dissolved iron upwelled and mixed with oxygenated surface water. The disappearance of iron formations after this event may reflect waning mafic-ultramafic magmatism and a diminished flux of hydrothermal iron relative to seawater oxidants.


Cytoplasmic RNA from normal and malignant human cells shows homology to the DNAs of Epstein-Barr virus and human adenoviruses.
Cytoplasmic RNA prepared from several human cell lines and tissues was hybridised to DNA from Epstein-Barr virus, human adenovirus types 2, 3 and 12 and human papovaviruses BK and JC. RNA from all the cells, regardless of whether or not they were virally infected, hybridised to specific regions of the Epstein-Barr virus or adenovirus genomes but not to papovavirus DNA. The cellular cross-hybridising species appear to be repetitive sequences which are conserved in higher eukaryotes. Mismatch estimations indicate a high degree of homology between the viral and host sequences. Detailed analysis of selected regions of viral DNA failed to reveal any primary-structural peculiarities.


Simultaneous trace determination of acidic non-steroidal anti-inflammatory drugs in purified water, tap water, juice, soda and energy drink by hollow fiber-based liquid-phase microextraction and ultra-high pressure liquid chromatography coupled to tandem mass spectrometry.
In this study, a two-phase hollow fiber liquid-phase microextraction (HF-LPME) coupling with ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed for determination of four non-steroidal anti-inflammatory (NSAIDs)-salicylic acid, ibuprofen, naproxen and diclofenac in real water samples. The influencing parameters of HF-LPME sample preparation method, such as organic solvents (acceptor phase), pH of sample solution (donor phase), extraction time, stirring speed, extraction temperature and ionic strength were systematically optimized. Through the developed determination method, high enrichment factors (195-346) were achieved for the four drugs. The instrumental calibration curves of salicylic acid, naproxen, diclofenac, and ibuprofen show good linear relations (R>0.998) in the concentration range of 1-500, 5-2500, 10-5000 and 5-2500 μg L(-1), respectively. The average recoveries of the four drugs in the low, medium and high spiked concentration levels (20-200, 50-500 and 100-1000 μg L(-1)) were between 98-115% with relative standard deviation (RSD) values were less than 12% (n=6). Limits of detection (LOD) of salicylic acid, naproxen, diclofenac, and ibuprofen in water were 0.5, 0.5, 1.0, and 1.25 μg L(-1), respectively. The determination method has been applied for the real samples (purified water, tap water, juice, soda and energy drinks), and the results show that salicylic acid was detected in tap water and soda, the concentrations were 2.85 μg L(-1) and 61.22 μg L(-1) separately, the RSD values were less than 9% (n=6). Salicylic acid and diclofenac were detected in energy drink, the concentrations were 44.62 μg L(-1) and 8.31 μg L(-1), the RSD values were less than 11% (n=6).


Inflammation, adiposity, and progression of arterial stiffness in adolescents with type 1 diabetes: The SEARCH CVD Study.
We examined the association between inflammation and progression of arterial stiffness in a population of youth with type 1 diabetes (T1D). A total of 287 youth with T1D (median age 13 years) from SEARCH CVD, an ancillary study to the SEARCH for Diabetes in Youth, were included. Markers of inflammation (CRP, IL-6, fibrinogen, leptin, and adiponectin) and measures of pulse wave velocity (PWV) of the arm (PWV-R), trunk (PWV-T), and lower extremity (PWV-LE) were measured at baseline. Measures of PWV were repeated approximately five years later. PWV-R (0.50 m/s), PWV-T (0.65 m/s), and PWV-LE (1.0 m/s) significantly increased over the follow-up (p < 0.001 for each). A significant interaction was found between waist circumference and fibrinogen (p = 0.036) on the progression of PWV-T, suggesting that fibrinogen is more strongly associated with PWV progression in lean participants. Improved understanding of adiposity, inflammation, and functional changes in the vascular system in patients with T1D is crucial.


The origin of the muscle fasciculation caused by funnel-web spider venom.
The origin of the fasciculation of skeletal muscle produced by funnel-web spider venom (FSV) has been examined in mouse phrenic nerve hemi-diaphragm preparations, FSV from male spiders at concentrations greater than 10(-6) g/ ml invariably produced muscle fasciculation which could be prevented by d-tubocurarine (14micron), tetrodotoxin (0.3 micron) or by increasing the external magnesium concentration or calcium concentration. Diphenyl hydantoin (3-6 X 10(-5) M) was able to reduce these fasciculations in some experiments. In curarized preparations, multiple end plate potentials (EPPs) in response to single stimuli and bursts of spontaneous EPPs were seen in the presence of FSV (10(-5) g/ml). Extracellular recordings from phrenic nerves in the presence of FSV (10(-5) g/ml) revealed additional components in compound action potentials elicited by single stimuli, and "spontaneous" electrical activity was observed in unstimulated nerves. This spontaneous activity was abolished by raising the divalent cation concentration in the bathing solution. These results suggest that a primary site of action of FSV is the surface membrane of nerve fibres and that muscle fasciculation arises as a consequence of spontaneous action potentials produced by the venom in motor nerves.


A fluorometric assay for acetylcholinesterase activity and inhibitor detection based on DNA-templated copper/silver nanoclusters.
A novel label-free, rapid, cost-effective, and highly sensitive fluorometric sensor has been constructed for the detection of acetylcholinesterase (AChE) activity and its inhibitor based on the fluorescence quenching of DNA-templated copper/silver nanoclusters (DNA-Cu/AgNCs). In this assay, AChE catalyzes the hydrolysis of acetylthiocholine (ATCh) to form thiocholine which induces fluorescence quenching of DNA-Cu/AgNCs. The AChE activity could be detected as low as 0.05mU/mL and with a linear range from 0.05 to 2.0mU/mL. This assay offers a very convenient "mix and detect" approach for AChE activity. On the other hand, tacrine and organophosphorus pesticides (OPPs) were employed to inhibit the hydrolysis of ATCh, which could eliminate the fluorescence quenching of DNA-Cu/AgNCs. The IC50 of tacrine and methamidophos were estimated to be 16.9nM and 0.075mg/L, respectively. This method was also used to detect spiked OPPs in agricultural products successfully. The present work may expand the use of DNA-Cu/AgNCs to the field of enzyme sensors.


Impact of Two Different Reference Measurement Procedures on Apparent System Accuracy of 18 CE-Marked Current-Generation Blood Glucose Monitoring Systems.
Measurement accuracy has been assessed for many different blood glucose monitoring systems (BGMS) over the years by different study groups. However, the choice of the comparison measurement procedure may impact the apparent level of accuracy found in such studies. Measurement accuracy of 18 different BGMS was assessed in a setting based on ISO 15197 using two different comparison methods in parallel: a glucose oxidase (GOD)-based and a hexokinase (HK)-based method. Accuracy limits of ISO 15197 were applied, and additional analyses were performed, including bias, linear regression, and mean absolute relative difference (MARD) to assess the impact of possible differences between comparison methods on the apparent level of accuracy. While ≈80% of BGMS met the accuracy criteria of ISO 15197 when compared with the respective manufacturers' reference measurement procedure, only two-thirds did so against both comparison methods. The mean relative bias ranged from -6.6% to +5.7% for the analysis against the GOD-based method and from -11.1% to +1.3% for the analysis against the HK-based method, whereas MARD results ranged from 3.7% to 9.8% and from 2.3% to 10.5%, respectively. Results of regression analysis showed slopes between 0.85 and 1.08 (GOD-based method) and between 0.81 and 1.01 (HK-based method). The results of this study indicate that there are systematic differences between the reference measurement procedures used for BGMS calibration as well as for system accuracy assessment. Because of the potential impact on therapy of patients with diabetes resulting from these differences, further steps toward harmonization of the measurement procedures' results are important.


Type 2 diabetes and pre-diabetic abnormalities in patients with bipolar disorders.
Abnormalities in the glucose metabolism cause nervous and organic damage and are a cardiovascular risk factor. They could be a main cause for the increased morbidity and mortality rates found in patients with bipolar disorders. The exact prevalence of diabetes and pre-diabetic abnormalities, however, is not clear. 85 euthymic outpatients with bipolar disorders from two university hospitals in Germany underwent an oral glucose tolerance test, laboratory screening and clinical measurements. Socio-demographic data, medication, severity of illness, global functioning and life quality were assessed. Diabetes mellitus was found in 7% of the patients, pre-diabetic abnormalities in 27%. The group of patients with abnormalities in the glucose metabolism had significantly lower quality of life and global functioning. Higher BMI, leptin, triglycerides and CRP levels significantly increased the likelihood for pre-diabetes/diabetes. The low sample size did only allow limited assessment of impact of medication on the results. No healthy controls were assessed. One-third of the patients with bipolar disorders showed abnormalities in the glucose metabolism and this was associated with impaired global functioning and lower quality of life. Early detection and intervention strategies fitting the needs of patient with bipolar disorder are needed to improve both physical and mental health.


Microbial community acclimatization for enhancement in the methane productivity of anaerobic co-digestion of fats, oil, and grease.
The methane productivity and long chain fatty acids (LCFAs) degradation capability of unacclimatized seed sludge (USS) and acclimatized seed sludge (ASS) at different substrate ratios of fats oil and grease (FOG) and mixed sewage sludge were investigated in this study. Biogas produced in ASS in initial phase of anaerobic digestion had higher methane content (65-76%) than that in USS (26-73%). The degradation of major LCFAs in the ASS was 22-80%, 33-191%, and 7-64% higher for the substrate ratios of 100:10, 100:20, and 100:30, respectively, as compared to the LCFAs' degradation in USS. Microbial acclimatization increased the population of Firmicutes (40%), Bacteroidetes (32%), Synergistetes (10%), and Euryarchaeota (8%) in ASS, which supported the faster rate of LCFAs degradation for its later conversion to methane. The significant abundance of Syntrophomonas and Methanosarcina genera in ASS supported faster generation rate of methane in an obligatory syntrophic relationship.


An investigation into whether nurse teachers take into account the individual learning styles of their students when formulating teaching strategies.
Many factors have an effect on ways students learn, but differences in learning styles may not be reflected in the way they are taught. A small study of the learning styles of 119 students and 13 nurse teachers was carried out as part of a Certificate of Education Course. In addition a short questionnaire was used to determine teachers preferred teaching styles. The data collected demonstrated that the groups sampled did not express a very strong preference for any one learning style, but for a combination of styles as did the teachers sampled. The author concludes that nurse teachers should examine more closely the reasons why they favour one strategy over another. This in turn should make learning more challenging and rewarding for the students and teachers.


How deep is the potential well confining a protein in a specific conformation? A single-molecule study on temperature dependence of conformational change between 5 and 18 K.
The fluorescence excitation spectrum of a single chromophore molecule in a photosynthetic pigment-protein complex is known to change in time at liquid helium temperature. The spectral change reflects a conformational change of the protein to which the chromophore binds. This work follows the temporal behavior of the spectrum of a single chromophore in the temperature range between 5 adn 18 K. The temperature dependence reveals two types of conformational change of the protein, i.e., thermally activated motions over a potential barrier of ca. 0.1 kJ/mol and temperature-independent motions of tunneling of a proton.


Neither glutamate nor alanine but arginine sensitizes BY-2 cells to arsenate.
Arsenic is toxic for plants. Our previous results showed that the application of proline enhanced the sensitivity of tobacco BY-2 cells to arsenate. In order to clarify the enhancement mechanism, we investigated the effects of other amino acids on the arsenate-stressed BY-2 cells. Glutamate at up to 10 m m did not affect the cell growth in the absence or presence of arsenate. Arginine at up to 10 m m did not affect the growth in the absence of arsenate but arginine at 10 m m enhanced the inhibition of the cell growth by arsenate. Alanine at up to 10 m m did not affect the cell growth under nonstressed condition but alanine at 10 m m significantly improved the cell growth under arsenate stress. These results suggest that alanine mitigates arsenate stress in BY-2 cells and that arginine like proline enhances the sensitivity of BY-2 cells to arsenate.


Provincial mortality in South Africa, 2000--priority-setting for now and a benchmark for the future.
Cause-of-death statistics are an essential component of health information. Despite improvements, underregistration and misclassification of causes make it difficult to interpret the official death statistics. To estimate consistent cause-specific death rates for the year 2000 and to identify the leading causes of death and premature mortality in the provinces. Total number of deaths and population size were estimated using the Actuarial Society of South Africa ASSA2000 AIDS and demographic model. Cause-of-death profiles based on Statistics South Africa's 15% sample, adjusted for misclassification of deaths due to ill-defined causes and AIDS deaths due to indicator conditions, were applied to the total deaths by age and sex. Age-standardised rates and years of life lost were calculated using age weighting and discounting. Life expectancy in KwaZulu-Natal and Mpumalanga is about 10 years lower than that in the Western Cape, the province with the lowest mortality rate. HIV/AIDS is the leading cause of premature mortality for all provinces. Mortality due to pre-transitional causes, such as diarrhoea, is more pronounced in the poorer and more rural provinces. In contrast, non-communicable disease mortality is similar across all provinces, although the cause profiles differ. Injury mortality rates are particularly high in provinces with large metropolitan areas and in Mpumalanga. The quadruple burden experienced in all provinces requires a broad range of interventions, including improved access to health care; ensuring that basic needs such as those related to water and sanitation are met; disease and injury prevention; and promotion of a healthy lifestyle. High death rates as a result of HIV/AIDS highlight the urgent need to accelerate the implementation of the treatment and prevention plan. In addition, there is an urgent need to improve the cause-of-death data system to provide reliable cause-of-death statistics at health district level.


Akt2 knockout mitigates chronic iNOS inhibition-induced cardiomyocyte atrophy and contractile dysfunction despite persistent insulin resistance.
Increased levels of inducible nitric oxide synthase (iNOS) during cardiac stress such as ischemia-reperfusion, sepsis and hypertension may display both beneficial and detrimental roles in cardiac contractile performance. However, the precise role of iNOS in the maintenance of cardiac contractile function remains elusive. This study was designed to determine the impact of chronic iNOS inhibition on cardiac contractile function and the underlying mechanism involved with a special focus on the NO downstream signaling molecule Akt. Male C57 or Akt2 knockout [Akt2(-/-)] mice were injected with the specific iNOS inhibitor 1400W (2 mg/kg/d) or saline for 7 days. Both 1400W and Akt2 knockout dampened glucose and insulin tolerance without additive effects. Treatment of 1400W decreased heart and liver weights as well as cardiomyocyte cross-sectional area in C57 but not Akt2 knockout mice. 1400W but not Akt2 knockout compromised cardiomyocyte mechanical properties including decreased peak shortening and maximal velocity of shortening/relengthening, prolonged relengthening duration, reduced intracellular Ca(2+) release and decay rate, the effects of which were ablated or attenuated by Akt2 knockout. Akt2 knockout but not 1400W increased the levels of intracellular Ca(2+) regulatory proteins including SERCA2a and phospholamban phosphorylation. 1400W reduced the level of anti-apoptotic protein Bcl-2, the effect of which was unaffected by Akt2 knockout. Neither 1400W nor Akt2 knockout significantly affected ER stress, autophagy, the post-insulin receptor signaling Akt, GSK3β and AMPK, as well as the stress signaling IκB, JNK, ERK and p38 with the exception of elevated IκB phosphorylation with jointed effect of 1400W and Akt2 knockout. Taken together, these data indicated that an essential role of iNOS in the maintenance of cardiac morphology and function possibly through an Akt2-dependent mechanism.


An evaluation of the contouring abilities of medical dosimetry students for the anatomy of a prostate cancer patient.
Prostate cancer is one of the most common diseases treated in a radiation oncology department. One of the major predictors of the treatment outcome and patient side effects is the accuracy of the anatomical contours for the treatment plan. Therefore, the purpose of this study was to determine which anatomical structures are most often contoured correctly and incorrectly by medical dosimetry students. The author also wanted to discover whether a review of the contouring rules would increase contouring accuracy. To achieve this, a male computed tomography dataset consisting of 72 transverse slices was sent to students for contouring. The students were instructed to import this dataset into their treatment planning system and contour the following structures: skin, bladder, rectum, prostate, penile bulb, seminal vesicles, left femoral head, and right femoral head. Upon completion of the contours, the contour file was evaluated against a "gold standard" contour set using StructSure software (Standard Imaging, Inc). A review of the initial contour results was conducted and then students were instructed to contour the dataset a second time. The results of this study showed significant differences between contouring sessions. These results and the standardization of contouring rules should benefit all individuals who participate in the treatment planning of cancer patients.


The cholinergic innervation of the rat cerebral cortex shows two distinct phases in development.
The cholinergic innervation of the rat cerebral cortex was examined in pre- and postnatal life using immunohistochemistry with a monoclonal antibody directed against choline acetyltransferase (ChAT). Our observations show that there are two separate phases in the development of the cholinergic innervation of the rat neocortex. The first, a transient phase, occurs in the late stages of gestation and in the perinatal period. During this time, ChAT-labelled cells (neuroblasts, as well as immature pyramidal and non-pyramidal neurons) are present throughout the entire rostro-caudal extent of the primordial cortex. The fate of these cells, which are not visible shortly after birth, is unknown as is their functional role in the developing cortex. The second phase in the development of the cholinergic innervation begins in the middle of the second postnatal week. At this stage only a few faintly stained neurons and fibres appear in the cortex. Their numbers and staining intensity increase gradually until the fifth postnatal week when ChAT-labelled neurons and axonal arbours appear indistinguishable from their adult counterparts. The pattern of development observed in the second phase parallels closely that shown in a recent analysis of cortical ChAT activity during postnatal life.


Plasma transforming growth factor beta1, metalloproteinase-1 and tissue inhibitor of metalloproteinases-1 in acute viral hepatitis type B.
Antiproliferative, pro-apoptotic and immunosuppressive activity effects suggest crucial role of transforming growth factor (TGF)-beta1, metalloproteinase (MMP)-1 and its tissue inhibitor (TIMP)-1 in the pathogenesis of acute liver injury that in some patients precede development of chronic liver diseases and fibrogenesis. The aim of this study was to evaluate effect of acute HBV infection on plasma TGF-beta1, MMP-1 and TIMP-1 levels. TGF-beta1, MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 39 patients with acute viral hepatitis type B. Baseline measurement was performed within the first week of jaundice and then weekly up to the fourth week of the disease. Results were compared to baseline and normal values and to liver function tests. Plasma concentrations of TGF-beta1, TIMP-1 and MMP-1 were significantly elevated in the first week of acute viral B hepatitis in comparison to normal. Analysis of individual values demonstrated significant positive correlation between plasma concentrations of TGF-beta1 and TIMP-1. There was no correlation between MMP-1 and TGF-beta1 or TIMP-1. Significant correlation was demonstrated between both TGF-beta1 and ALT or AST as well as between TIMP-1 and ALT, AST or bilirubin. Elevated baseline levels of both TGF-beta1 and TIMP-1 decreased gradually in consecutive weeks of the disease. TGF-beta1 but not TIMP-1 plasma concentrations were significantly lower in 3rd and 4th week than baseline values. MMP-1 concentration remained on baseline level in the 2nd week of the disease. However in the 3rd week its values increased suddenly but the significant difference in comparison to baseline was observed only in 4th week. These results indicate important role of TGF-beta1, TIMP-1 and MMP-1 in acute viral hepatitis, that seems to be connected first of all with hepatocytes damage. Their role in extracellular matrix metabolism during acute liver injury needs further evaluation.


Transluminal angioplasty of isolated crural arterial lesions in diabetics with critical limb ischemia.
We investigated the long-term clinical results of transluminal angioplasty of isolated infrapopliteal arteries in diabetic patients with severe ischaemic foot lesions and the influence of different parameters on primary success, the long-term outcome and the survival rate. Between January 2001 and May 2004 we performed 50 transluminal angioplasties in 52 (10 female, 42 male, mean age 69.3 years) diabetics with isolated lesions of crural arteries (tibio-peroneal trunk 41, anterior tibial 13, posterior tibial 5, peroneal artery 14) and limb threatening ischaemia (rest pain 12, tissue loss 40). In two patients the dilatation could not be performed due to vessel perforation and impossibility to catheter localization. The morphological classification of infrapopliteal lesions was determined according to guidelines provided by TransAtlantic Inter-Society Consensus (TASC) (17 type A, 16 type B, 13 type C, 4 type D). Initial technical success after angioplasty of crural arteries could be obtained in 96%. Kaplan-Meier analysis showed a cumulative limb salvage of 92%, 85% and 68,9% after 6 months, one and two years, respectively. Patients falling in TASC group C had significantly higher risk of amputation. Overall survival at I year was calculated at 85% and at 2 years at 65%. Our results suggest that depending on the extent of lesions transluminal angioplasty of infrapopliteal artery stenoses and occlusions is considered as an effective and save therapy modality to avoid limb loss in diabetics with critical ischemia.


Cleavage of territrems by alkaline hydrogen peroxide: Isolation and characterization of the aromatic moiety of territrems.
The chemical reaction of cleavaging territrem B to give 3,4,5-trimethoxy benzoic acid by alkaline hydrogen peroxide was investigated. The method was applied for confirmation of the chemical structure of the aromatic moiety of territrem A, A', B, and B'. The physicochemical properties of the aromatic cleavage product of territrem Aindicated the structure as 3,4-methylendioxy, 5-methoxy benzoic acid (or 4-methoxy, 6-carboxy, 1, 3-benzodioxole). The experiment also gave the evidences that territrem A and A', on the other hand territrem B and B' have the identical aromatic moieties on their structures.


Replication regions of Sinorhizobium meliloti plasmids.
The replication (rep) regions of small plasmids from three Sinorhizobium meliloti strains were cloned by marker rescue. Two unique replication regions were identified, one of which was common to two different strains. Plasmid pBB83 carried a 7.2 kbp rep region from a 42 kbp plasmid, and pBB84 carried a 4.5 kbp rep region from a 36 kbp plasmid. The cloned rep regions were of different compatibility types, and were capable of displacing their parent plasmids from S. meliloti. Neither could function in a PolA- strain of Escherichia coli. The cloned replication regions were less stable in S. meliloti than their parent plasmids. The rep genes for each plasmid were localized to less than 2.5 kbp segments. Sequencing data revealed that the pBB83 Rep protein is uncommon, with partial identity to a protein encoded by a plasmid from S. meliloti GR4 [Mercado-Blanco, J., Olivares, J., 1994. The large nonsymbiotic plasmid pRmeGR4a of Rhizobium meliloti GR4 encodes a protein involved in replication that has homology with the RepC protein of Agrobacterium plasmids. Plasmid 32, 75-79]. However, the cloned DNA fragment also contains a truncated segment of the common repABC genes, suggesting that the parent plasmid contained two sets of replication genes. Other genes and an IS-element within the insert are most closely related to sequences derived from the Rhizobiaceae family, suggesting that the plasmid has a limited host range. In contrast, the pBB84 rep region contained genes similar to those associated with several broad host-range plasmids, and its Rep protein is related to that of a Pseudomonas aeruginosa broad host-range plasmid, pVS1 [Heeb, S., Itoh, Y., Nishijyo, T., Schnider, U., Keel, C., Wade, J., Walsh, U., O'Gara, F., Haas, D., 2000. Small, stable shuttle vectors based on the minimal pVS1 replicon for use in gram-negative, plant-associated bacteria. Mol. Plant-Microbe Interact. 13, 232-237]. The pBB84 rep region also includes a probable origin of replication, consisting of DNA boxes flanking a series of direct repeats and an AT-rich sequence.


BAF180 is a critical regulator of p21 induction and a tumor suppressor mutated in breast cancer.
Screening for tumor suppressor genes in breast cancer revealed multiple truncating mutations of PB1, which encodes the BAF180 subunit of the PBAF chromatin remodeling complex. Mutation was associated with loss of heterozygosity of the wild-type allele. BAF180 complementation of BAF180-mutant tumor cells caused G(1) arrest that was dependent on increased expression of the cyclin/cyclin-dependent kinase inhibitor p21/WAF1/CIP1. Endogenous wild-type BAF180 bound to the p21 promoter and was required for proper p21 expression and G(1) arrest after transforming growth factor-beta and gamma-radiation treatment. BAF180 thus functions on two tumor suppressor signaling pathways as a physiologic mediator of p21 expression. We conclude that BAF180 suppresses tumorigenesis, at least in part, through its ability to regulate p21.


Objective and subjective diagnostic parameters in the fellow eye of unilateral keratoconus.
Keratoconus (KC) is usually a bilateral corneal ectatic disease. For significant asymmetric presentation (so called unilateral KC), the fellow eye has the mildest and earliest form of the disease, which is typically called forme fruste keratoconus. The aim of this study was to evaluate the sensitivity and specificity of parameters derived from a Scheimpflug imaging system (Pentacam) as well as the changes in the quality of mesopic vision in the apparently normal fellow eye (forme fruste) to detect the earliest and most sensitive parameters. Patients with clinical keratoconus in one eye and forme fruste keratoconus in the fellow eye were compared to subjects with normal eyes. The patients were examined using a rotating Scheimpflug imaging system (Pentacam).The following parameters were evaluated: keratometry, minimum corneal thickness, pachymetry progression index (PPI), Ambrósio relational thickness (ART), posterior elevation, back difference elevation (BDE) and multimetric D index(D index). Receiver operating characteristic (ROC) curves were analyzed by evaluating the area under the curve (AUC) to detect the sensitivity and specificity of each parameter. Mesopic vision evaluations were performed by contrast sensitivity and glare tests for each group. A total of 48 patients with clinical keratoconus in one eye and forme fruste keratoconus in the fellow eye and 72normal subjects were evaluated. In the clinical keratoconus eyes, the mean K, back difference elevation (BDE), pachymetric progression index maximum(PPI max), and multimetric D were significantly higher compared to the normal subjects, whereas the corneal pachymetry and Ambrósio relational thickness maximum (ART max) were significantly lower. In the forme fruste eyes, the ROC analysis showed that the AUC values of the mean K, thinnest pachymetry, ARTmax, BDE, D index, and PPI max were 0.82, 0.61, 0.88, 0. 67, and 0.64, respectively. The contrast sensitivity and glare tests were significantly affected in the forme fruste cases. In forme fruste keratoconus eyes, the ART max is considered a highly sensitive objective parameter. Contrast sensitivity and glare is an important subjective test, which is affected in forme fruste patients.


LC and LC-MS methods for the investigation of polypills for the treatment of cardiovascular diseases. Part 1. Separation of active components and classification of their interaction/degradation products.
"Polypill" is a fixed-dose combination (FDC) containing three or more drugs in a single pill. The same is under development for the treatment and prevention of cardiovascular diseases. In the present study, gradient LC methods were developed for simultaneous determination of the possible components of a polypill, i.e., lisinopril, aspirin and one each among atenolol/hydrochlorothiazide and atorvastatin/simvastatin/pravastatin, in the presence of a total of 13 major interaction/degradation products. The drugs and the products were well separated using a reversed-phase (C-8) column and a mobile phase comprising of acetonitrile: phosphate buffer (pH 2.3). Other HPLC parameters were flow rate, 1 ml/min; detection wavelength, 210 nm; column oven temperature, 60 degrees C; and injection volume, 5 microl. The methods were validated for linearity, precision, accuracy, and specificity. These were further modified to make them compatible for LC-MS studies by removal of the phosphate buffer and adjustment of pH by formic acid. The suitability of the methods for LC-MS studies was established by matching the theoretical mass values of the drugs with those obtained experimentally. These methods were used to determine mass values of the major interaction/degradation products, which helped to know the source of their origin.


Altered expression of cyclooxygenase-2, presenilins and oxygen radical scavenging enzymes in a rat model of global perinatal asphyxia.
Cyclooxygenase-2 (COX-2) and presenilin (PSEN) 1 and 2 genes are regulated during development as well as in pathological conditions associated to hypoxia. In this study, we investigated their patterns of expression during the first 2 weeks of postnatal life in a rat model of moderate global perinatal asphyxia that we have previously reported to be characterized by early oxidative stress and delayed behavioral alterations. In the hippocampus, global perinatal asphyxia induced an early up-regulation COX-2 mRNA (postnatal day, pnd, 1), which preceded those of PSEN 1 and 2 genes, observed at pnd 4. At pnd 11, the expression of all three genes was decreased compared to control animals. In addition, we analyzed the expression of the scavenging enzymes catalase, copper/zinc- and manganese-superoxide dismutase. As for COX-2, the three enzymes were up-regulated in the hippocampus at pnd 1 and returned at or below baseline by pnd 4. In the cortex, only PSEN 1 and 2 showed a moderate up-regulation at pnd 1 followed by a down-regulation at pnd 11. These findings suggest that moderate perinatal hypoxic episodes are associated with a dysregulation of the several genes involved in brain development and anti-oxidant defenses, which follows a rapid and transient oxidative stress. Such alterations are particularly evident in the hippocampus and could represent an adaptive response to the hypoxic condition. The delayed down-regulation of the scavenging enzymes could set the ground for brain damage and delayed behavioral alterations and further support the potential benefit of early anti-oxidant treatments in a short therapeutic window soon after birth.


Myoadenylate deaminase deficiency: absence of correlation with exercise intolerance in 452 muscle biopsies.
A histochemical assay was routinely performed of myoadenylate deaminase (MAD) in muscle biopsy specimens. MAD was absent in 13 cases, i.e. 2.9% of the specimens. In 10 cases the deficiency was confirmed biochemically. The diagnoses in the 13 patients were: polyneuropathy (n = 5), infantile spinal muscular atrophy (n = 3), congenital myopathy with type 2 fibre atrophy, facioscapulohumeral myopathy, polymyositis, myotonic dystrophy and hyperornithinaemia with gyrate atrophy of the retina. In contrast, 35 unrelated patients presenting with exercise-related muscle cramps or pains showed normal histochemical MAD activity. The biopsy specimens in all of these patients were essentially normal and in none of them was the diagnosis of a neuromuscular disease made. The results failed to confirm the association of MAD deficiency with aches, cramps and pains or exertional myalgia.


Lethal oxidative damage to human immunodeficiency virus by human recombinant myeloperoxidase.
Human recombinant myeloperoxidase was evaluated in a cell-free system for its inactivation properties on the replication of human immunodeficiency virus, HTLV-IIIB. In the presence of a hydrogen peroxide generating system (glucose and glucose oxidase) and sodium thiocyanate, the recombinant enzyme inhibited virus-induced syncytium formation and viral replication without causing any cytopathic effects on SupT1 reporter cells. In addition, U937 monocytoid cells, chronically infected with HIV1, were exposed to recombinant myeloperoxidase (10 U/ml) and monitored during 48 h for the accumulation of intracellular p24 viral antigen. Under these conditions, the recombinant enzyme significantly reduced intracellular viral replication without affecting cell viability.


Light transmission in negative curvature hollow core fiber in extremely high material loss region.
In this paper we demonstrate the light transmission in a spectral range of 2.5 to 7.9 µm through a silica negative curvature hollow core fiber (NCHCF) with a cladding consisting of eight capillaries. A separation between the cladding capillaries was introduced to remove the additional resonances in the transmission bands. The measured optical loss at 3.39 µm was about 50 dB/km under a few modes waveguide regime.


Photosensitive seizures provoked while viewing "pocket monsters," a made-for-television animation program in Japan.
To describe the recent epidemic of photosensitive seizure that occurred in relation to an episode of the television animation program "Pocket Monsters," we report four patients who experienced seizures while watching the episode in question. We also report some technical aspects of the program episode. We investigated the clinical symptoms of the four patients and performed routine EEGs with intermittent photic stimulation (IPS). If IPS provoked no photoparoxysmal response (PPR) during the routine EEG examination, a second EEG was performed with the photic stimulator placed 10 cm from the patient's eyes. In addition, we reviewed the "Pocket Monsters" episode, focusing our attention on the visual techniques used with reference to the Independent Television Commission (ITC) guidelines. One patient who had myoclonic jerks before the convulsion in question was diagnosed as having juvenile myoclonic epilepsy, and the diagnosis of another patient was pure photosensitive epilepsy. The remaining two patients had their first seizures, which could be occasional seizures, and we therefore could not reach a diagnosis of epilepsy. In our four patients, only one showed PPR on the routine EEG. Two patients revealed PPR on the second EEG, and the remaining patient showed no PPR. Rapid changes in color are believed to be responsible for the photosensitive seizures because all four patients had seizures at around 18:50, when seconds of deep red and bright blue flashes, alternating at a frequency of 12 Hz, were shown. Regulations for technical aspects of children's programming, including the use of colors, are urgently needed in Japan to prevent a repeated incident. In addition, the IPS procedure needs to be standardized, especially for patients who are suspected to have photosensitivity.


Physical Examination Sensitivity for Skull Fracture in Pediatric Patients With Blunt Head Trauma: A Secondary Analysis of the National Emergency X-Radiography Utilization Study II Head Computed Tomography Validation Study.
We evaluated the emergency department (ED) providers' ability to detect skull fractures in pediatric patients presenting with blunt head trauma. This was a secondary analysis of the National Emergency X-Radiography Utilization Study (NEXUS) Head computed tomography (CT) validation study. Demographics and clinical characteristics were analyzed for pediatric patients. Radiologist interpretations of head CT imaging were abstracted and cataloged. Detection of skull fractures was evaluated through provider response to specific clinical decision instrument criteria (NEXUS or Canadian head CT rules) at the time of initial patient evaluation. The presence of skull fracture was determined by formal radiologist interpretation of CT imaging. Between April 2006, and December 2015, 1,018 pediatric patients were enrolled. One hundred twenty-eight (12.5%) children had a notable injury reported on CT head. Skull fracture was present in most (66.4%) children with intracranial injuries. The sensitivity and specificity of provider physical examination to detect skull fractures was 18.5% (95% confidence interval 10.5% to 28.7%) and 96.6% (95.3% to 97.7%), respectively. The most common injuries associated with skull fractures were subarachnoid hemorrhage (27%) and subdural hematoma (22.3%). Skull fracture is common in children with intracranial injury after blunt head trauma. Despite this, providers were found to have poor sensitivity for skull fractures in this population, and these injuries may be missed on initial emergency department assessment.


Minor histocompatibility antigens are developmentally regulated on murine embryonal carcinoma cells and their early differentiated derivatives.
Differences in the expression of minor histocompatibility (Hm) alloantigens on two mouse embryonal carcinoma (EC) cell lines and the PYS-2 and T.D.M.-1 differentiated derivatives have been demonstrated by their ability to elicit a cytolytic T lymphocyte response. Experiments involving the use of various responder-target strain combinations and recombinant inbred mice strains have shown that: (1) there are major differences in Hm expression on EC cells compared with differentiated derivatives whose Hm expression appears more like that of adult splenocytes; (2) although both EC cell lines show reduced Hm immunogenicity compared with adult splenocytes, major differences in the expression and possible presentation of Hm between the F9 and PCC3 EC cell lines can be detected by in vivo priming and by in vitro cold competition target experiments. These observations are discussed in relation to the differences in allograft rejection patterns observed with PCC3 and F9 and to possible differences in developmental staging of these cell lines.


Determination of hydrogen sulfide and volatile thiols in air samples by mercury probe derivatization coupled with liquid chromatography-atomic fluorescence spectrometry.
A new procedure is proposed for the sampling and storage of hydrogen sulphide (H2S) and volatile thiols (methanethiol or methyl mercaptan, ethanethiol and propanethiol) for their determination by liquid chromatography. The sampling procedure is based on the trapping/pre-concentration of the analytes in alkaline aqueous solution containing an organic mercurial probe p-hydroxymercurybenzoate, HO-Hg-C6H4-COO- (PHMB), where they are derivatized to stable PHMB complexes based on mercury-sulfur covalent bonds. PHMB complexes are separated on a C18 reverse phase column, allowing their determination by liquid chromatography coupled with sequential non-selective UV-vis (DAD) and mercury specific (chemical vapor generation atomic fluorescence spectrometry, CVGAFS) on-line detectors. PHMB complexes, S(PHMB)2CH3S-PHMB, C2H5S-PHMB and C3H7S-PHMB, are stable alt least for 12 h at room temperature and for 3 months if stored frozen (-20 degrees C). The best analytical figures of merits in the optimized conditions were obtained by CVGAFS detection, with detection limits (LODc) of 9.7 microg L(-1) for H2S, 13.7 microg L(-1) for CH(3)SH, 17.7 microg L(-1) for C2H5SH and 21.7 microg L(-1) for C3H7SH in the trapping solution in form of RS-PHMB complexes, the relative standard deviation (R.S.D.) ranging between 1.0 and 1.5%, and a linear dynamic range (LDR) between 10 and 9700 microg L(-1). Conventional UV absorbance detectors tuned at 254 nm can be employed as well with comparable R.S.D. and LDR, but with LODc one order of magnitude higher than AFS detector and lower specificity. The sampling procedure followed by LC-DAD-CVGAFS analysis has been validated, as example, for H2S determination by a certified gas permeation tube as a source of 3.071+/-0.154 microg min(-1) of H2S, giving a recovery of 99.8+/-7% and it has been applied to the determination of sulfur compounds in real gas samples (biogas and the air of a plant for fractional distillation of crude oil).


Combination therapy with isosorbide dinitrate: current status and the future.
The excellent safety and predictable efficacy of isosorbide dinitrate (ISDN) have been demonstrated repeatedly during the past 25 years in a number of studies in which the agent has been used alone or in combination with other antianginal agents. Clinical studies to investigate the additive or synergistic effect of ISDN have been difficult to conduct because of the complexity of protocol design and length of studies required. However, combination therapy is well accepted in the clinical practice of medicine and cardiology and is used to obtain additive therapeutic effects while minimizing the side effects. The addition of ISDN not only to other standard and proven antianginal agents but also to calcium antagonists should prove to be a fruitful area for further clinical research benefiting patients with angina pectoris (caused by either coronary artery spasm or occlusive coronary artery disease), hypertension, and congestive heart failure. Noncardiovascular uses of ISDN may include the treatment of hyperspasticity of other smooth muscle beds, such as esophageal spasm and achalasia.


[Effect of prolonged cold storage and subsequent heat treatment on the fractional composition of fish muscle proteins].
The effect of prolonged preservation at -18 degrees C and subsequent culinary heat treatment of the fractional composition of sarcoplasmatic and myofibrillar proteins in the muscles of mirror carp, pike and sheat-fish were studied by the method of gel-filtration on Sephadex G-100. Cold storage of the dish at -18 degrees C for 9 months was found to produce market changes in the qualitative and quantitative composition of muscle proteins. After culinary heat treatment the finished products have lost much of their original quality.


Safety of anticoagulation use for treatment of portal vein thrombosis in liver cirrhosis and its effect on hospital-based outcomes: an insight from a US nationwide database.
Anticoagulation use for portal vein thrombosis (PVT) in patients with advanced liver disease is controversial. We investigated the effect of anticoagulation on outcomes in patients with PVT with cirrhosis. We reviewed National Inpatient Sample data from 2016 to 2018 to identify patients with PVT. Our outcomes were in-hospital mortality, variceal bleeding, hepatic encephalopathy, acute kidney injury (AKI), hepatorenal syndrome (HRS), spontaneous bacterial peritonitis (SBP), sepsis and hospital resource utilisation. We included 60 505 patients with PVT, out of whom 6.63% (4015) were on anticoagulation. The overall mortality in the anticoagulation group was 2.12% compared with 9.72% in the no anticoagulation group. The adjusted odds of mortality were low in the anticoagulation group (adjusted OR (AOR) 0.27, 95% CI 0.15 to 0.46, p<0.001). Patients on anticoagulation had 29% lower odds of variceal bleeding (AOR 0.71, 95% CI 0.53 to 0.96, p=0.03). Lower odds of HRS (AOR 0.56, 95% CI 0.37 to 0.85, p=0.01) and AKI (AOR 0.57, 95% CI 0.48 to 0.69, p<0.001) were also seen in the anticoagulation group. Patients in the anticoagulation group also showed lower odds of SBP (AOR 0.62, 95% CI 0.43 to 0.89, p=0.01) and sepsis (AOR 0.57, 95% CI 0.35 to 0.93, p=0.03). Anticoagulation use resulted in shorter hospital stay by 1.15 days (adjusted length of stay -1.15, 95% CI -1.51 to -0.79, p<0.001). The mean difference in total hospital charges between the anticoagulation and the no anticoagulation group was -$20 034 (95% CI -$27 077 to -$12 991, p<0.001). Our analysis found that anticoagulation use is safe and associated with better outcomes in patients with PVT with advanced liver disease.


Economic and resource status of the chiropractic profession in Ontario, Canada: a challenge or an opportunity.
Chiropractic is one of the most frequently sought nonphysician provider groups. Despite its apparent recognition, the profession faces numerous challenges, including the economic reality of an increasing supply within a market of questionable demand. This paper evaluates the chiropractic manpower status in Ontario, Canada. Data collected from administrative and education databases, insurance billing data, and population health survey data between 1990 and 2004 were analyzed. Between 1990 and 2004, the total number of chiropractic registrants in Ontario doubled, with an average annual rate of growth of about 5.4%; however, recent data suggest that the number of nonpracticing chiropractors is increasing, whereas the number of new registrants is decreasing. The rate of applications to a chiropractic institution rose sharply and peaked in 1996-1997, thereafter declining but leveling off in 2002-2003. Despite the continued growth in the number of practicing chiropractors, the utilization of chiropractic services among the Ontario population has remained relatively stable, resulting in a decline in the average net annual incomes adjusted for inflation to 2002 dollars. Our results support previous reports projecting an oversupply of chiropractors and suggest that the chiropractic profession in Ontario is in long-run oversupply. Competition from other providers, changing population demographics, and the recent loss of public funding for services may present significant future challenges to current practitioners. Opportunities related to participation in multidisciplinary environments and accessing unmet population health needs may contribute to influencing the demand for chiropractic services. A concerted effort by professional and educational institutions is required.


Comparison of spinal anesthesia with combined sciatic-femoral nerve block for outpatient knee arthroscopy.
To compare spinal anesthesia and combined sciatic-femoral nerve block for outpatient knee arthroscopy. Prospective, randomized, controlled study. Postoperative recovery area at a university-affiliated medical center. 50 ASA physical status I and II adult outpatients undergoing arthroscopic knee surgery. Study subjects were equally divided (n = 25 each) into spinal and sciatic-femoral groups. Spinal group patients received spinal anesthesia with 7.5 mg of 0.5% hyperbaric bupivacaine. Sciatic-femoral group patients received combined sciatic-femoral nerve blocks using a mixture of 20 mL of lidocaine 2% plus 20 mL of bupivacaine 0.5%. Times including that from arrival in the operating room to readiness for surgery, duration of surgery, recovery time, and patient satisfaction were recorded. Analgesia and occurrence of adverse events also were recorded. No significant differences between the two groups were found for any of the study measurements of recovery. After discharge, postoperative pain differed significantly between groups only at 6 hours (P < 0.002). Patient satisfaction was high with both techniques. Combined sciatic-femoral nerve block for outpatient arthroscopic knee surgery offers satisfactory anesthesia, with a clinical profile similar to that of low-dose spinal anesthesia. Sciatic-femoral nerve blocks are associated with significantly lower pain scores during the first 6 postoperative hours.


Cerebral ischaemia in pituitary disorders--more common than previously thought: two case reports and literature review.
Vasospasm and consequent cerebral ischaemia in aneurysmal subarachnoid haemorrhage are well-described. The development of cerebral ischaemia following pituitary tumour surgery is under-appreciated, and can be attributed to mainly cerebral vasospasm or internal carotid artery compression. We report on two patients with pituitary tumours who developed delayed cerebral ischaemia after transsphenoidal and transcranial pituitary macroadenoma decompression. The patients had vasospasm of intracranial vessels demonstrable on magnetic resonance angiogram. One recovered neurologically following nimodipine and hypertensive-hypervolaemia therapy while the other developed fulminant cerebral infarction. We discuss the complex multi-factorial mechanisms of cerebral ischaemia in pituitary disorders, as well as the management strategies and their limitations.


Structure and function of the transcription elongation factor GreB bound to bacterial RNA polymerase.
Bacterial GreA and GreB promote transcription elongation by stimulating an endogenous, endonucleolytic transcript cleavage activity of the RNA polymerase. The structure of Escherichia coli core RNA polymerase bound to GreB was determined by cryo-electron microscopy and image processing of helical crystals to a nominal resolution of 15 A, allowing fitting of high-resolution RNA polymerase and GreB structures. In the resulting model, the GreB N-terminal coiled-coil domain extends 45 A through a channel directly to the RNA polymerase active site. The model leads to detailed insights into the mechanism of Gre factor activity that explains a wide range of experimental observations and points to a key role for conserved acidic residues at the tip of the Gre factor coiled coil in modifying the RNA polymerase active site to catalyze the cleavage reaction. Mutational studies confirm that these positions are critical for Gre factor function.


Effect of systemic photobiomodulation in the course of acute lung injury in rats.
Acute lung injury (ALI) is a severe, multifactorial lung pathology characterized by diffuse alveolar injury, inflammatory cell infiltration, alveolar epithelial barrier rupture, alveolar edema, and impaired pulmonary gas exchange, with a high rate of mortality; and sepsis is its most common cause. The mechanisms underlying ALI due to systemic inflammation were investigated experimentally by systemic lipopolysaccharide (LPS) administration. Photobiomodulation (PBM) has been showing good results for several inflammatory diseases, but there are not enough studies to support the real benefits of its use, especially systemically. Considering that ALI is a pathology with high morbidity and mortality, we studied the effect of systemic PBM with red light-emitting diode (LED) (wavelength 660 nm; potency 100 mW; energy density 5 J/cm; total energy 15 J; time 150 s) in the management of inflammatory parameters of this disease. For this, 54 male Wistar rats were submitted to ALI by LPS injection (IP) and treated or not with PBM systemically in the tail 2 and 6 h after LPS injection. Data were analyzed by one-way ANOVA followed by Student's Newman-Keuls. Our results point to the beneficial effects of systemic PBM on the LPS-induced ALI, as it reduced the number of neutrophils recruited into the bronchoalveolar lavage, myeloperoxidase activity, and also reduced interleukins (IL) 1β, IL-6, and IL-17 in the lung. Even considering the promising results, we highlight the importance of further studies to understand the mechanisms involved, and especially the dosimetry, so that in near future, we can apply this knowledge in clinical practice.


DNA barcoding reveals ongoing immunoediting of clonal cancer populations during metastatic progression and immunotherapy response.
Cancers evade the immune system through the process of cancer immunoediting. While immune checkpoint inhibitors are effective for reactivating tumour immunity in some cancer types, many other solid cancers, including breast cancer, remain largely non-responsive. Understanding how non-responsive cancers evade immunity and whether this occurs at the clonal level will improve immunotherapeutic design. Here we use DNA barcoding to track murine mammary cancer cell clones during immunoediting and determine clonal transcriptional profiles that allow immune evasion following anti-PD1 plus anti-CTLA4 immunotherapy. Clonal diversity is significantly restricted by immunotherapy treatment in both primary tumours and metastases, demonstrating selection for pre-existing breast cancer cell populations and ongoing immunoediting during metastasis and treatment. Immunotherapy resistant clones express a common gene signature associated with poor survival of basal-like breast cancer patient cohorts. At least one of these genes has an existing small molecule that can potentially be used to improve immunotherapy response.


A PCR survey for posterior Hox genes in amphibians.
Hox genes encode transcription factors that play a key role in specifying the body plan in metazoans and are therefore essential in explaining patterns of evolutionary diversity. As an ancient tetrapod group with diverse limb types, amphibians are important for understanding the origin and diversification of limbs in land vertebrates. We conducted a PCR survey in two species of each amphibian order to identify Hox-9 to Hox-13, known to function in limb development. Fifteen distinct posterior Hox genes and one retro-pseudogene were identified, and the former confirm the existence of four Hox clusters in each amphibian order. Some genes expected to occur in all tetrapods, based on the posterior Hox complement of mammals, fishes and coelacanth, were not recovered from our survey, and may have been lost. Hoxd-12 is absent in frogs and possibly other amphibians. Considering its function in autopodial development, the loss of this gene may be related to the absence of the fifth finger in frogs and salamanders.


Inhibition of rat heart and liver microsomal lipid peroxidation by nifedipine.
Lipid peroxidation in rat heart and liver microsomes was induced by an NADPH-generating system or by ascorbate in the presence of an ADP-iron complex. Microsomal lipid peroxidation, as measured by malonaldehyde formation, was inhibited by nifedipine over a wide range of concentrations (47 microM to 6 mM). Nifedipine also decreased the oxygen consumption of cardiac and hepatic microsomes in a concentration-dependent manner. These results indicate that nifedipine may perturb microsomal electron transport systems. Nifedipine may have the potential to alter the sensitivity of cardiac and hepatic membranes to peroxidative damage.


Complementarity of colestipol, niacin, and lovastatin in treatment of severe familial hypercholesterolemia.
To compare the effectiveness of the ternary-drug combination of colestipol, niacin, and lovastatin with binary combinations of those drugs in treating patients with familial hypercholesterolemia. An open sequential study of serum lipoprotein responses in patients receiving diet alone (mean duration, 4 months); colestipol and niacin with diet (mean duration, 9 months); and colestipol, niacin, and lovastatin with diet (mean duration, 15 months). Metabolic ward and lipid clinic of a university medical center. Twenty-two patients with clinical characteristics of familial hypercholesterolemia (low-density-lipoprotein cholesterol, greater than 8.48 mmol/L; 21 of 22 with tendon xanthomas). Diet: less than 200 mg/d of cholesterol and less than 8% of total calories from saturated fat; colestipol, 30 g/d; lovastatin, 40 to 60 mg/d; and niacin, 1.5 to 7.5 g/d. Mean total serum cholesterol and low-density-lipoprotein cholesterol levels of 4.86 +/- 0.62 mmol/L (188 +/- 24 mg/dL SD) and 2.89 +/- 0.54 mmol/L (112 +/- 21 mg/dL SD), respectively, were significantly lower during ternary-drug treatment than during colestipol-niacin treatment (p less than 0.003) or during treatment in which other possible binary combinations were given. The cholesterol content of very low-density-lipoproteins was lower and high-density-lipoprotein cholesterol levels higher during this phase than during the colestipol-niacin phase. Colestipol, lovastatin, and niacin are mutually complementary in treating hypercholesterolemia. This regimen produces reductions in serum cholesterol levels similar to those associated with regression of atheromatous plaques in animal studies.


Epigenetic regulation of myofibroblast differentiation and extracellular matrix production in nasal polyp-derived fibroblasts.
Nasal polyposis is a multi-factorial disease associated with chronic inflammatory condition of the paranasal sinuses. Myofibroblast differentiation and extracellular matrix (ECM) accumulation are involved in the pathogenesis of nasal polyposis. The aim of this study was to study the effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on transforming growth factor (TGF)-β1-induced myofibroblast differentiation and ECM accumulation in nasal polyp-derived fibroblasts (NPDFs). Nasal polyp-derived fibroblasts were isolated from nasal polyps of patients who have chronic rhinosinusitis with nasal polyp. TSA was treated in TGF-β1-induced NPDFs. Expression levels of HDAC2, α-smooth muscle actin (SMA), TGF-β1, collagen type I, acetylated Histone H3, acetylated Histone H4, phosphorylated Smad2/3 and Smad7 were determined by RT-PCR, western blot and/or immunofluorescent staining. The total collagen amount production was analysed by Sircol soluble collagen assay and contractile activity was measured by collagen gel contraction assay. HDAC2 inhibition by TSA or HDAC2 silencing was established by RT-PCR and western blot. The epigenetic effect on α-SMA gene inactivation was examined by chromatin immunoprecipitation assay. Proliferation was determined by Ki67-positive cell staining and cytotoxicity was assessed by 3-(4,5- dimethylthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The expression levels of HDAC2, α-SMA and TGF-β1 were increased in nasal polyp tissues compared to normal inferior turbinate tissues. TSA and HDAC2 silencing inhibited expression levels α-SMA, collagen and HDAC2. TSA induced hyperacetylation of histone and suppressed opening of α-SMA gene promoter in TGF-β1-induced NPDFs. TSA inhibited TGF-β1-induced Smad 2/3 and rescued TGF-β1-suppressed Smad7 signalling pathway. Finally, TSA blocked proliferation in TGF-β1-induced NPDFs and has no cytotoxic effect in NPDFs. These results suggest that HDAC inhibition is associated with myofibroblast differentiation and extracelluar matrix accumulation in nasal polyposis. TSA may be useful as an inhibitor of nasal polyp growth, and thus has potential to be used as a novel treatment option for nasal polyposis.


Deficiency of CD73 activity promotes protective cardiac immunity against Trypanosoma cruzi infection but permissive environment in visceral adipose tissue.
Damaged cells release the pro-inflammatory signal ATP, which is degraded by the ectonucleotidases CD39 and CD73 to the anti-inflammatory mediator adenosine (ADO). The balance between ATP/ADO is known to determine the outcome of inflammation/infection. However, modulation of the local immune response in different tissues due to changes in the balance of purinergic metabolites has yet to be investigated. Here, we explored the contribution of CD73-derived ADO on the acute immune response against Trypanosoma cruzi parasite, which invades and proliferates within different target tissues. Deficiency of CD73 activity led to an enhanced cardiac microbicidal immune response with an augmented frequency of macrophages with inflammatory phenotype and increased CD8+ T cell effector functions. The increment of local inducible nitric oxide (NO) synthase (iNOS)+ macrophages and the consequent rise of myocardial NO production in association with reduced ADO levels induced protection against T. cruzi infection as observed by the diminished cardiac parasite burden compared to their wild-type (WT) counterpart. Unexpectedly, parasitemia was substantially raised in CD73KO mice in comparison with WT mice, suggesting the existence of tissue reservoir/s outside myocardium. Indeed, CD73KO liver and visceral adipose tissue (VAT) showed increased parasite burden associated with a reduced ATP/ADO ratio and the lack of substantial microbicidal immune response. These data reveal that the purinergic system has a tissue-dependent impact on the host immune response against T. cruzi infection.


Formal and Informal Services Used by Women Who Suffer Intimate Partner Violence in Spain.
Intimate partner violence (IPV) is an important public health problem. Female victims of IPV do not always use the institutional resources available to them. We conducted a cross-sectional study using data from the Spanish 2014 Macro-Survey on Violence Against Women. The findings show that 15% of the survey participants suffered from IPV in the last year. The factors associated with a higher probability of suffering IPV are being of non-Spanish nationality, being a student, having a low-educational level, having no income, being the household head, and not having a current partner. Regarding the resources used by women subjected to IPV, almost half resorted to informal sources, such as female friends and/or their own mothers. The use of formal resources was low. Therefore, IPV continues to be a problem in Spain that seems to remain in the private domain. Consequently, it is necessary to increase the availability of and access to legal resources.


[Food allergy in the child: an exploratory study on the impact of the elimination diet on food neophobia].
This study was designed to analyse the impact of an elimination diet in children with food allergy, and its perception by their parents on the later reticence of children to test unknown foods, food neophobia. The degree of food neophobia of children having outgrown their allergy (mean age, 7 years 2 months) was compared to that of a sibling (9 years 5 months) using a standardized scale and a questionnaire of food friendliness. Parents were also asked to fill in a questionnaire on the disease and its burden on the family. Children having outgrown their allergy are more reluctant to test new foods than their non-allergic brother or sister, as shown by their scoring on the food neophobia scale and the number of unknown foods following the cure of the disease. Two factors increase the level of food neophobia, the distressing effect and the duration of the period elapsed until the diagnosis was made, as well as the distressing effect and the lack of variety in the meal preparation. Food neophobia, a normal phase between 2 and 10 years, is worsened by the elimination diet required by food allergy, especially in case of late diagnosis and when the time elapsed before diagnosis and the preparation of meals were perceived as difficult to bear.


Nutritional Status of Children from Women with Previously Bariatric Surgery.
Number of pregnancies has been increasing in women of childbearing age after the gastric bypass. The objective of this study was to evaluate the nutritional status of children of women submitted to gastric bypass. We evaluated anthropometric, breastfeeding and biochemical profile, body composition, and dietary intake indicators of children of both sexes who were born alive after the surgery. For statistical analysis, were performed Shapiro-Wilk and ANOVA test (p < 0.05). The sample consisted of 13 children (61.6% female, mean age of 46 ± 22.3 months, BMI of 18.9 ± 3.3 kg/m2). The classification of BMI index by age showed that 46.1% of the children were normal weight and 30.8% obese. We observed a large percentage of children with deficiency of iron and vitamin A. 7.6 and 30.7% of children presented carbohydrate and lipid, respectively, lower than the recommendation. Fiber intake was inadequate in all children, calcium in 61.5%, vitamin A in 30.7%, and folate in 76.9% of them. Also, 84.6% presented sodium intake higher than the recommendations. The blood glucose levels were lower in children with maternal breastfeeding (65.5 ± 2.1 mg/dL, p < 0.05); furthermore, children breastfed with artificial and breast milk presented lower fat mass (3.8 ± 1.9 kg; p < 0.05). Children from women with previously gastric bypass presented low birth weight; however, they are currently underweight or overweight and present important deficiency of iron and vitamin A and inadequate alimentary intake mainly of sodium and fibers. Breastfeeding may play a protective role in the development of obesity in these children.


Lactobacillus salivarius CECT 5713, a potential probiotic strain isolated from infant feces and breast milk of a mother-child pair.
In this study, Lactobacillus salivarius CECT 5713 was originally isolated from feces of a one-month-old breast-fed infant. Since it has been suggested that the gut microbiota of breast-fed infants reflects that of the maternal breast milk, we investigated if this specific strain was present in breast milk of the respective mother. RAPD and PFGE analysis revealed the presence of the strain L. salivarius CECT 5713 in this biological fluid. To our knowledge, this is the first report of a L. salivarius strain isolated from breast milk. L. salivarius CECT 5713 produced l-lactate, acetate and hydrogen peroxide, which may be responsible for its antimicrobial activity against most of the indicator organisms used in this study; in addition, this strain showed a high survival rate after exposition to conditions simulating those found in the gastrointestinal tract. Finally, it was strongly adhesive to Caco-2 and HT-29 cells did not produce biogenic amines and were unable to degrade gastric mucin in vitro.


Large congenital melanocytic nevus. Light and electron microscopic findings.
The family of a 10-month-old black girl with a large periocular congenital melanocytic nevus (CMN) was given markedly differing advice from experienced physicians regarding how the lesion should be managed. The nevus was eventually removed surgically because of concern about the risk of malignant transformation and because of its cosmetic appearance. Histologically, the lesion was a dermal nevus, but extended deeply into subcutaneous tissue and had neuroid features. The ultrastructural characteristics of melanocytes varied according to the depth at which they resided, but cells did not differ basically from melanocytes found in smaller acquired nevi. Given the limited understanding of the biologic basis for malignant transformation, large size remains the most important risk factor in CMN.


Chronic hepatitis E in an elderly immunocompetent patient who achieved a sustained virologic response with ribavirin treatment.
A woman in her late 70 s was diagnosed with liver injury at a health examination. Despite treatment with ursodeoxycholic acid at a nearby hospital, her transaminase levels elevated in two peaks. She was transferred to our hospital 77 days after the health examination. She weighed 42 kg and had a low body mass index of 19.8 kg/m2. Viral markers, including immunoglobulin A (IgA) against hepatitis E virus (anti-HEV IgA), were negative. Drug-induced liver injury was negligible. We suspected autoimmune hepatitis because of the patient's female gender and positive antinuclear antibody. However, prednisolone and azathioprine failed to completely improve her hepatitis. On day 643, anti-HEV IgA was re-evaluated and found to be positive. She was diagnosed with autochthonous chronic hepatitis E because the virus strains in the preserved serum on day 77 and the serum on day 643 had identical nucleotide sequences (genotype 3a). Following prednisolone and azathioprine discontinuation, ribavirin (RBV) was administered for 3 months. HEV RNA disappeared and remained negative for more than 6 months after the cessation of RBV. The HEV RNA titer of 6.2 log10 copies/mL on day 77 was unusually high 2.5 months after the onset, suggesting that hepatitis E had already been chronic before immunosuppressive treatment for possible autoimmune hepatitis. After getting married at 23 years old, she had been a housewife and had no comorbidities that might deteriorate her immunity. Chronicity should be kept in mind when encountering HEV infection in elderly and underweight patients.


The Role of Therapeutic Drug Monitoring in Children.
The use of biologic therapies has changed the treatment landscape for children with inflammatory bowel disease. While the novel biologics have improved clinical outcomes, there remains a significant gap in achieving endoscopic remission, prolonged steroid-free remission, and drug durability. Contributing to this gap is the paucity of real-world pharmacokinetic studies in children and a failure to dose optimize therapy during induction. Emerging data from a pediatric clinical trial and several observational studies have shown that the combination of proactive therapeutic drug monitoring and achievement of early therapeutic concentrations is effective in achieving improved outcomes. The next steps will be to leverage these past studies to develop more innovative clinical trials to properly assess the safety and effectiveness of proactive therapeutic drug monitoring in children.


[Joint and functional changes in the masticatory system in chronic rheumatoid arthritis in adults. Preliminary study].
A clinical and radiological study has been carried out to highlight signs and symptoms of mandibular dysfunction in a sample of 66 adult patients suffering from chronic rheumatoid arthritis. Data were quantified on the basis of Helkimo indices. Prevalence proved significantly high, particularly as regards joint crepitus, reduced mandibular mobility, pain in the masticatory muscles and macroglossia. It did not seem to be linked to age. Rather it presented a definite predominance of the female sex, confirmed by comparison with a sample of nonrheumatic controls. Contemporaneous X-ray examination showed a bilateral location, in many cases remodelling and reabsorption of the condyle, an increase in the articular space and erosion of the fossa, at times reduction in the space and sclerosis, dependent on a superimposed degenerative arthrosis (usually more painful). Generally speaking, however, there was a coincidence between severity of clinical dysfunction and seriousness of the morphological lesions whereas subjective symptoms were relatively independent. The need for polyspecialistic cooperation and a multidisciplinary therapeutic approach is reiterated.


Silver positivity of the NORs during embryonic development of Xenopus laevis.
Transcriptional activity of ribosomal RNA (rRNA) genes is detectable around blastula-gastrula transition during the embryonic development of amphibians and other non-mammalian systems. The silver staining reaction, known to selectively stain transcriptionally active nucleolus organizer regions (NORs) both in interphase and metaphase chromosomes allowed us to follow the activation of the NORs during the embryonic development of Xenopus laevis.


[Clinical value of induced sputum test in monitoring airway inflammation in asthma].
To explore the clinical value of induced sputum test in monitoring airway inflammation in asthmatic patients. The induced sputum test was done in 12 normal subjects and 21 asthmatic subjects; The levels of interleukin-8 (IL-8), eosinophil cation protein (ECP), malondialdehyde (MDA) in plasma and induced sputum in all subjects were measured. The levels of IL-8, ECP, MDA in plasma and induced sputum in unstable asthmatic patients were significantly higher than those in stable asthmatic (P < 0.01), in which the levels of IL-8, ECP, MDA in both plasma and induced sputum were significantly higher than those in normal subjects(P < 0.05); In all asthmatic patients, the levels of IL-8, ECP, MDA in induced sputum were significantly higher than those in plasma. The levels of IL-8, ECP, MDA in induced sputum could be used for monitoring airway inflammation in asthmatic patients.


Expression of all six human Torque teno virus (TTV) proteins in bacteria and in insect cells, and analysis of their IgG responses.
Torque teno virus (TTV) is a non-enveloped human virus with a circular ( approximately 3800 nt) ssDNA genome. TTV transcription results in three viral mRNAs and six proteins, the function or antigenicity of which are unknown. The six open reading frames of TTV genotype 6 were expressed in bacteria and insect cells. Expression of the ORF1/1-encoded protein was inefficient, while expression of the others was successful, with ORF1 and ORF1/2 as arginine-rich region depleted. All six recombinant TTV proteins were antigenic. Of healthy adults, 11/25 (44%) showed strong IgG reactivity with one or more proteins. Four subjects, two of whom were genotype-6-DNA positive, were followed. One of the latter showed concurrently a strong IgG response against the ORF1 protein. The other showed appearance of IgG against the ORF2 protein concomitantly with resolution of the genotype-6 viremia. The genotype-6 sequences remained unaltered for years, suggesting that some mechanisms other than amino acid substitutions play a role in TTV immune evasion.


A philosophy for big-bang cosmology.
According to recent developments in cosmology we seem bound to find a model universe like the observed universe, almost independently of how we suppose it started. Such ideas, if valid, provide fresh justification for the procedures of current cosmological theory.


Electrostatic complexes of mitomycin C with nucleic acids and polyanions.
Reductively activated mitomycin C exhibits strong, non-covalent electrostatic binding to polyanions such as polyvinylsulfate and polyphosphate. The protonated C-2 amino group generated by the reduction is most likely responsible for this type of interaction. At moderate drug and salt concentrations only covalent binding to nucleic acids is observable. This is shown to be guanine-specific in DNA for the first time, as well as in synthetic polyribo- and polydeoxyribonucleotides at 10--20 times higher binding levels than previously tested. At higher mitomycin C concentration, however, strong non-covalent electrostatic binding to nucleic acids also occurs, resulting in a binding ratio up to 1 mol drug bound per mol mononucleotide, although this non-specific binding is relatively inhibited compared to polyvinylsulfate. Salts also have an inhibitory effect on the non-specific binding to nucleic acids. A series of mitomycin derivatives were compared for their binding and cross-linking abilities using DNA as substrate, with the following results: (a) the presence of a basic nitrogen . funtion at C-2 promotes binding, both covalent and electrostatic, presumably by kinetically facilitating the approach between positively charged nitrogen and DNA. (b) The aziridine ring is the major covalent binding site, indispensable for crosslinking and determines the guanine-specificity of the binding.


Differential associative behaviour of mitotic and meiotic acrocentric chromosomes.
A comparative study of the association of mitotic acrocentric chromosomes and acrocentric bivalents at the pachytene stage shows that at least two factors can act in the associative behaviour of these chromosomes: (1) Nor activity and (2) the presence of satellite DNA in the short arms of these chromosomes. These factors do not act with the same intensity in the two cell lines studied. In lymphocytes, Nor activity prevails, whereas satellite DNA plays the main role in the association of acrocentric chromosomes in germ cells at the pachytene stage.


Safety of antithrombotic drugs in patients with atrial fibrillation and non-end-stage chronic kidney disease: Meta-analysis and systematic review.
To evaluate the safety of antithrombotic drugs used in patients with both atrial fibrillation (AF) and non-end-stage chronic kidney disease (NECKD). A search was performed for studies on major bleeding outcomes in patients with concurrent AF and NECKD using Medline and Cochrane databases on 19th February, 2015. Fixed- or random-effects meta-analysis was adopted for evaluating pooled effect sizes according to whether heterogeneity existed. Twelve articles were included for analysis. Three studies evaluated AF patients who took warfarin vs. placebo/antiplatelet drugs in the presence of NECKD. No significant difference in major bleeding risk was observed according to the pooled analysis using the random-effects model (RR: 1.05, 95% CI: 0.74-1.36). The risk of a composite of major bleeding outcomes was reduced by 19% in patients randomized to direct oral anticoagulants (DOACs) compared to dose-adjusted warfarin from pooled data of three randomized controlled trials with regard to AF and NECKD (RR: 0.81, 95% CI: 0.75-0.88). This superiority of DOACs to warfarin maintained until the renal function was severely impaired. In patients with AF and NECKD, no significant increase in the incidence of major bleeding outcomes was observed in warfarin use compared with placebo/antiplatelet drugs. DOACs reduced the risk of major bleeding by 19% compared to warfarin and further data-exploration indicated that the risk did not increase as renal function deteriorated during the renal status of mild to moderate impairment.


Peripheral corticotropin-releasing factor induces diarrhea in rats: role of CRF1 receptor in fecal watery excretion.
Systemic injection of corticotropin-releasing factor (CRF) stimulates colonic secretory and motor functions, and CRF receptors play a role in stress-related alterations of colonic functions. Stress has also been reported to induce diarrhea and we investigated if peripheral injection of CRF can mimic this response in conscious rats. Intravenous (i.v.) injection of CRF (3, 10 or 30 microg/kg) caused diarrhea in 13%, 63% and 75% of rats, respectively, and dose dependently increased the fecal fluid content by 5.1-, 8.6- and 10.8-fold, while the dried solid weight was increased by 5.2-, 4.9- and 5.8-fold, respectively, compared to the i.v. saline group. CRF actions were rapid in onset and blocked by the CRF1 receptor, antagonist CP-154,526 (butyl-[2,5-dimethyl-7-(2,4,6-trimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]ethylamine). These results demonstrate that peripheral CRF induces watery diarrhea, primarily through the activation of CRF1 receptor suggesting a possible role for these pathways in colonic responses to stress.


MRI in Lower Extremity Peripheral Arterial Disease: Recent Advancements.
Evaluation of peripheral arterial disease by cardiovascular magnetic resonance imaging continues to develop. Of the clinical diagnostics tests currently available, magnetic resonance angiography is well established as one of the preferred techniques for determining areas of arterial occlusive disease affecting the lower extremities. Despite this, there have been new developments in non-gadolinium based contrast-enhanced studies as well as testing done at higher field strength scanners. In the research arena, magnetic resonance spectroscopy, calf muscle perfusion imaging and atherosclerotic plaque evaluation all have made significant advancements over the last year. These techniques are gaining traction as surrogate endpoints in clinical trials of novel therapeutics aimed at alleviating symptoms in patients with peripheral arterial disease.


Microbial Resistance Mechanisms to the Antibiotic and Phytotoxin Fusaric Acid.
Fusaric acid (FA) produced by Fusarium oxysporum plays an important role in disease development in plants, including cotton. This non-specific toxin also has antibiotic effects on microorganisms. Thus, one expects a potential pool of diverse detoxification mechanisms of FA in nature. Bacteria and fungi from soils infested with Fusarium and from laboratory sources were evaluated for their ability to grow in the presence of FA and to alter the structure of FA into less toxic compounds. None of the bacterial strains were able to chemically modify FA. Highly FA-resistant strains were found only in Gram-negative bacteria, mainly in the genus of Pseudomonas. The FA resistance of the Gram-negative bacteria was positively correlated with the number of predicted genes for FA efflux pumps present in the genome. Phylogenetic analysis of predicted FA resistance proteins (FUSC, an inner membrane transporter component of the efflux pump) revealed that FUSC proteins having high sequence identities with the functionally characterized FA resistance protein FusC or Fdt might be the major contributors of FA resistance. In contrast, most fungi converted FA to less toxic compounds regardless of the level of FA resistance they exhibited. Five derivatives were detected, and the detoxification of FA involved either oxidative reactions on the butyl side chain or reductive reactions on the carboxylic acid group. The production of these metabolites from widely different phyla indicates that resistance to FA by altering its structure is highly conserved. A few FA resistant saprophytic or biocontrol strains of fungi were incapable of altering FA, indicating a possible involvement of efflux transporters. Deployment of both efflux and derivatization mechanisms may be a common feature of fungal FA resistance.


Relationship between serial serum neutrophil-lymphocyte ratio, cardiovascular mortality, and all-cause mortality in Chinese peritoneal dialysis patients.
Background It is believed that the excessive cardiovascular (CV) burden of patients on peritoneal dialysis (PD) is closely associated with chronic inflammation. Neutrophil-lymphocyte ratio (NLR) is an inflammatory marker that was shown to correlate with CV outcomes. However, little is known about the significance of serial monitoring of serum NLR. We aimed to determine the prognostic value of serial NLR on all-cause mortality and CV mortality in PD patients. Methods Serial measurement of NLR was obtained from 225 incident PD patients in a single center, with each measurement one year apart. Patients were divided into two groups ('high' versus 'low') by the median value of NLR. The primary and secondary outcome measure was all-cause and CV mortality respectively. Results After a median of follow up for 43.9 months, patients with lower baseline NLR demonstrated a higher survival rate (p=0.01). Patients with persistently high NLR values on serial measurement had the lowest survival rate (p=0.03). Multivariate Cox regression showed that this group of patients had significantly higher all-cause mortality (HR 1.74, 95%CI 1.09 to 2.79, p=0.02). However, the NLR failed to demonstrate a statistically significant relationship with CV mortality. Conclusions While baseline NLR was an independent predictor of all-cause mortality in PD patients, persistent elevation in NLR appeared to further amplify the risk. Regular monitoring of serial serum NLR may enable early identification of patients who are at risk of adverse outcome.


A biotin-avidin enzyme immunoassay for the quantitation of serum ferritin.
A biotin-avidin enzyme immunoassay (EIA) for measurement of ferritin in human serum is described. This procedure involves the use of specific antibody immobilized on beads, biotin-labeled specific antibody and enzyme-labeled avidin. Reproducible results were achieved within 2 h for ferritin in serum in the range of 5-500 ng/ml. Ferritin levels as low as 0.1 ng/ml can be measured. The biotin-avidin EIA, a radioimmunoassay kit, and an EIA kit were used to determine serum ferritin levels in healthy adults. Good agreement was found between the biotin-avidin EIA and the other methods.