Ethmoid sinus cancer: results of treatment with surgery and combined therapy.
Ethmoid sinus cancer is a rare paranasal sinus malignancy. Its characteristics include a low incidence rate, a great variety of histopathological types and multiple treatment modalities. Currently, there remains no definite consensus regarding its optimal management. The aim of this study was to examine the outcome of a population of Asian patients with advanced ethmoid sinus cancers that had been treated with surgery plus combined therapy. Between January 1989 and December 2002 inclusive, 19 newly diagnosed patients with ethmoid sinus cancers who had undergone surgical intervention were enrolled, T4 being the principal carcinoma stage (68.4%). All participating cases proved to be node-negative and no evidence of any distant metastasis was detected at the time of diagnosis. The major treatment modality was surgery plus postoperative radiotherapy. All but 2 of the 13 patients with T4 cancer underwent craniofacial resection with pericranial flap reconstruction. The estimated overall and disease-free survival rates 3 years post-treatment were 49.4% and 26.3%, respectively. Local tumor recurrence was more common than regional recurrence and/or distant metastasis. A total of 5/15 T3-T4 patients (33%) developed a neck metastasis, 3 of whom also suffered a distant metastasis. There was no postoperative mortality for the cases treated with craniofacial resection. Ethmoid sinus cancer typically demonstrates a propensity for late diagnosis and poor prognosis. This study confirms that craniofacial resection plus combined associated therapy is the optimal approach for the effective management of extensive ethmoid sinus tumors and is associated with an acceptable morbidity rate. More aggressive disease management featuring prophylactic concurrent chemoradiotherapy including neck or elective neck dissection plus chemotherapy should be considered for T3-T4 patients as opposed to T1-T2 patients.


Changing role for preoperative embolisation in the management of arteriovenous malformations of the brain.
The aim of this study was to analyse the results of the use of preoperative embolisation in the management of arteriovenous malformations of the brain at one institution between 1989 and 1999. Two hundred and fifty consecutive cases of angiographically confirmed arteriovenous malformations underwent surgery by one surgeon. Cases of dural or spinal arteriovenous malformations have been excluded. Forty-five cases underwent preoperative embolisation. Embolisation was mostly by particulate embolic material delivered 4 to 6 days before the intended surgery. The incidence of embolisation declined from 21 cases of the first 50 arteriovenous malformation cases surgically treated to five in the last 50 cases. For arteriovenous malformations of less than 3 cm, only the first two temporal quintiles had embolised cases; six in the first and three in the second. Outcome was measured by the Modified Rankin Scale. By 12 months (or last follow up, if less than this time has elapsed) following surgery, 1.6% of patients had died, 2.4% had a Modified Rankin scale score of 4 or 5, 6.4% had a Modified Rankin scale score of 3, 8.4% had a Modified Rankin scale score of 2, 14.4% had a Modified Rankin scale score of 1, and 66.8% were without neurological deficit. There was no difference in outcomes in each of the temporal quintiles. The four deaths were related to intraoperative haemorrhage, ruptured aneurysm, acute myocardial infarction or unrelated infection. Angiographic cure was achieved in 244 of 246 surviving cases. The two cases with residual arteriovenous malformations underwent focussed irradiation. Permanent morbidity could be attributable to embolisation, intraoperative events (resection functional brain, arteriovenous malformation rupture, aneurysm rupture or myocardial infarction) or postoperative events (arterio-capillary-venous hypertensive syndrome or infection). Of these 29 patients 14 had undergone embolisation. Mortality and major morbidity (Modified Rankin scale score greater than 2 due to treatment) occurred in 8.8% undergoing embolisation compared with 1.9% not embolised. The cause for major morbidity in these four embolised cases was intraoperative or postoperative haemorrhage. These results reflect that cases selected for embolisation were those at most risk from intraoperative haemorrhage. Arteriovenous malformations that are less than 3 cm in maximal diameter should only rarely be considered for preoperative embolisation because of their low surgical morbidity. In the presence of a significant deep perforating artery contribution that cannot be effectively embolised the risks of operative haemorrhage is high irrespective of the effectiveness of embolising ancillary non-perforating arteries.


Laboratory screening in the assessment of human cataract.
Cataracts may clearly arise in association with systemic metabolic disorders, but most ophthalmologists do not request an extended laboratory assessment for patients presenting with cataracts who are otherwise in good health. This approach was not unreasonable in view of the very low yield of unsuspected abnormalities revealed by detailed screening, but should be reconsidered in view of recent advances particularly in the understanding of partial defects of galactose metabolism. Proposals for a laboratory screening policy in relation to metabolic cataracts are discussed.


Arterial spin-labeled perfusion of pediatric brain tumors.
Pediatric brain tumors have diverse pathologic features, which poses diagnostic challenges. Although perfusion evaluation of adult tumors is well established, hemodynamic properties are not well characterized in children. Our goal was to apply arterial spin-labeling perfusion for various pathologic types of pediatric brain tumors and evaluate the role of arterial spin-labeling in the prediction of tumor grade. Arterial spin-labeling perfusion of 54 children (mean age, 7.5 years; 33 boys and 21 girls) with treatment-naive brain tumors was retrospectively evaluated. The 3D pseudocontinuous spin-echo arterial spin-labeling technique was acquired at 3T MR imaging. Maximal relative tumor blood flow was obtained by use of the ROI method and was compared with tumor histologic features and grade. Tumors consisted of astrocytic (20), embryonal (11), ependymal (3), mixed neuronal-glial (8), choroid plexus (5), craniopharyngioma (4), and other pathologic types (3). The maximal relative tumor blood flow of high-grade tumors (grades III and IV) was significantly higher than that of low-grade tumors (grades I and II) (P < .001). There was a wider relative tumor blood flow range among high-grade tumors (2.14 ± 1.78) compared with low-grade tumors (0.60 ± 0.29) (P < .001). Across the cohort, relative tumor blood flow did not distinguish individual histology; however, among posterior fossa tumors, relative tumor blood flow was significantly higher for medulloblastoma compared with pilocytic astrocytoma (P = .014). Characteristic arterial spin-labeling perfusion patterns were seen among diverse pathologic types of brain tumors in children. Arterial spin-labeling perfusion can be used to distinguish high-grade and low-grade tumors.


To grow old: regulatory role of ethylene and jasmonic acid in senescence.
Senescence, the final stage in the development of an organ or whole plant, is a genetically programmed process controlled by developmental and environmental signals. Age-related signals underlie the onset of senescence in specific organs (leaf, flower, and fruit) as well as the whole plant (monocarpic senescence). Rudimentary to most senescence processes is the plant hormone ethylene, a small gaseous molecule critical to diverse processes throughout the life of the plant. The role of ethylene in senescence was discovered almost 100 years ago, but the molecular mechanisms by which ethylene regulates senescence have been deciphered more recently primarily through genetic and molecular studies in Arabidopsis. Jasmonic acid (JA), another plant hormone, is emerging as a key player in the control of senescence. The regulatory network of ethylene and JA involves the integration of transcription factors, microRNAs, and other hormones. In this review, we summarize the current understanding of ethylene's role in senescence, and discuss the interplay of ethylene with JA in the regulation of senescence.


Long-term treatment with 3,4-methylenedioxymethamphetamine caused retina damage in C57BL/6 mice.
As a class of psychostimulant drugs, amphetamines are widely abused for their stimulant, euphoric, and hallucinogenic properties. Many of these effects result from acute increases in dopamine and serotonin neurotransmission. Following the onset of these effects, 3,4 methylenedioxymethamphetamine produces persistent damage to dopamine and serotonin nerve terminals, resulting in long-lasting neurotoxicity. The purpose of this investigation was to assess the effects of treatment with low dose of methylenedioxymethamphetamine on retinal function of C57BL/6 mice and its underlying mechanisms. C57BL/6 mice were divided randomly into two groups (n=10): one group was treated with phosphate buffered saline by intraperitoneal injection daily; the other group was treated with 1 mg/kg methylenedioxymethamphetamine by intraperitoneal injection daily for three months. Electroretinography was used to test retinal function every month. H&E staining and terminal deoxynucleotidyl transferase assay were used to evaluate the retinal morphology and histology. Enzyme-linked immunosorbent assay assays were used to measure markers of oxidative stress and inflammatory factors. Gene and protein expression was detected by real-time PCR and western blot. Three-month treatment with methylenedioxymethamphetamine induced significant retinal dysfunction via photoreceptor cell apoptosis by oxidative stress and inflammatory responses. These results suggest that long-term treatment with methylenedioxymethamphetamine increases inflammatory responses in photoreceptor cells resulting in retinal dysfunction in C57BL/6 mice. Thus, this investigation provides preclinical rationale for the retina damage caused by the methylenedioxymethamphetamine abuse.


42S p48--the most abundant protein in previtellogenic Xenopus oocytes--resembles elongation factor 1 alpha structurally and functionally.
We have undertaken an immunological and biochemical analysis of the most abundant soluble protein of previtellogenic Xenopus oocytes, 42S p48. We show that this protein shares immunological cross-reactivity with elongation factor 1 alpha (EF-1 alpha). Direct assays of both 42S fractions and purified 42S p48 show that this cross-reactivity is of functional significance since 42S p48, like EF-1 alpha, can transfer charged amino acids to ribosomes. We further demonstrate that 42S p48 is degraded soon after the onset of vitellogenesis, while the EF-1 alpha concentration remains essentially unchanged during this transition. These properties of 42S p48 are discussed with regard to its role in oogenesis.


Interest in genetic testing for modest changes in breast cancer risk: implications for SNP testing.
Advances in genomics may eventually lead to 'personalized genetic medicine,' yet the clinical utility of predictive testing for modest changes in risk is unclear. We explored interest in genetic testing for genes related to modest changes in breast cancer risk in women at moderate to high risk for breast cancer. Women (n = 105) with a negative breast biopsy and ≥1 relative with breast or ovarian cancer completed telephone surveys. We measured demographic and psychosocial variables and, following presentation of hypothetical scenarios of genetic tests for lower-penetrance breast cancer gene mutations, assessed interest in willingness to pay for and comprehension of test results. We used logistic regression models with generalized estimating equations to evaluate combinations of risk level, cost and behavioral modifiers. Many women (77%) reported 'definite' interest in genetic testing, with greater interest in tests that conveyed more risk and cost less. Behavioral modifiers of risk (taking a vitamin; diet/exercise), having a regular physician, greater perceived benefits of genetic testing, and greater cancer worry also influenced interest. Most participants (63%) did not understand relative vs. absolute risk. Women with less understanding reported more cancer worry and greater willingness to pay for testing. Interest in genetic testing for mutations related to modest changes in risk was high, modified by both test and psychosocial factors. Findings highlight the need for education about benefits and risks of testing for mutations that convey modest changes in risk, particularly given the current lack of clinical validity/utility and availability of direct-to-consumer genetic testing.


Antimyelin antibodies as a predictor of clinically definite multiple sclerosis after a first demyelinating event.
Most patients with multiple sclerosis initially present with a clinically isolated syndrome. Despite the fact that clinically definite multiple sclerosis will develop in up to 80 percent of these patients, the course of the disease is unpredictable at its onset and requires long-term observation or repeated magnetic resonance imaging (MRI). We investigated whether the presence of serum antibodies against myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) in patients with a clinically isolated syndrome predicts the interval to conversion to clinically definite multiple sclerosis. A total of 103 patients with a clinically isolated syndrome, positive findings on cerebral MRI, and oligoclonal bands in the cerebrospinal fluid were studied. At base line, serum samples were collected to test for anti-MOG and anti-MBP antibodies with Western blot analysis, and the lesions detected by cerebral MRI were quantified. Neurologic examinations for relapse or disease progression (defined as conversion to clinically definite multiple sclerosis) were performed at base line and subsequently every three months. Patients with anti-MOG and anti-MBP antibodies had relapses more often and earlier than patients without these antibodies. Only 9 of 39 antibody-seronegative patients (23 percent) had a relapse, and the mean (+/-SD) time to relapse was 45.1+/-13.7 months. In contrast, 21 of 22 patients (95 percent) with antibodies against both MOG and MBP had a relapse within a mean of 7.5+/-4.4 months, and 35 of 42 patients (83 percent) with only anti-MOG antibodies had a relapse within 14.6+/-9.6 months (P<0.001 for both comparisons with antibody-seronegative patients). The adjusted hazard ratio for the development of clinically definite multiple sclerosis was 76.5 (95 percent confidence interval, 20.6 to 284.6) among the patients who were seropositive for both antibodies and 31.6 (95 percent confidence interval, 9.5 to 104.5) among the patients who were seropositive only for anti-MOG antibodies, as compared with the seronegative patients. Analysis of antibodies against MOG and MBP in patients with a clinically isolated syndrome is a rapid, inexpensive, and precise method for the prediction of early conversion to clinically definite multiple sclerosis. This finding may be important for the counseling and care of patients with a first demyelinating event suggestive of multiple sclerosis.


Endoscopic guided intubation of long nasointestinal decompression tube.
Surgeons have long been faced with the problem of intubating long nasointestinal tubes through the pylorus of the stomach when intestinal obstructions are present. The procedure described herein provides a direct view of intubation, endoscopically, rather than a blind approach fluoroscopically. This is achieved in every instance through the use of a guided wire inserted through an endoscope with a sleeve type, long nasogastrointestinal decompression tube. The procedure has proved to be advantageous because of the time saved by providing for rapid passage and prompt decompression which also eliminates trips and calls to the radiology department. Also, the improved condition of the patient prior to operation and the decreased amount of radiation that the patient is exposed to are advantages.


A fully implantable device for intraperitoneal drug delivery refilled by ingestible capsules.
Creating fully implantable robots that replace or restore physiological processes is a great challenge in medical robotics. Restoring blood glucose homeostasis in patients with type 1 diabetes is particularly interesting in this sense. Intraperitoneal insulin delivery could revolutionize type 1 diabetes treatment. At present, the intraperitoneal route is little used because it relies on accessing ports connecting intraperitoneal catheters to external reservoirs. Drug-loaded pills transported across the digestive system to refill an implantable reservoir in a minimally invasive fashion could open new possibilities in intraperitoneal delivery. Here, we describe PILLSID (PILl-refiLled implanted System for Intraperitoneal Delivery), a fully implantable robotic device refillable through ingestible magnetic pills carrying drugs. Once refilled, the device acts as a programmable microinfusion system for precise intraperitoneal delivery. The robotic device is grounded on a combination of magnetic switchable components, miniaturized mechatronic elements, a wireless powering system, and a control unit to implement the refilling and control the infusion processes. In this study, we describe the PILLSID prototyping. The device key blocks are validated as single components and within the integrated device at the preclinical level. We demonstrate that the refilling mechanism works efficiently in vivo and that the blood glucose level can be safely regulated in diabetic swine. The device weights 165 grams and is 78 millimeters by 63 millimeters by 35 millimeters, comparable with commercial implantable devices yet overcoming the urgent critical issues related to reservoir refilling and powering.


An adjunctive method of radiofrequency volumetric tissue reduction of the tongue for OSAS.
Temperature-controlled radiofrequency volumetric reduction (TCRF), a minimally invasive procedure, has been used to treat tongue base obstruction in Obstructive Sleep Apnea Syndrome (OSAS). An adjunctive method was objectively evaluated. A prospective, nonrandomized clinical study was undertaken on 20 consecutive OSAS patients with isolated tongue base obstruction. Under local anesthesia, multiple lesions of the ventral tongue (genioglossus insertion) and dorsal tongue were given at each treatment session. A visual analog scale was used to assess changes in speech and swallowing. Polysomnography and Epworth Sleepiness Scale (ESS) were used to assess outcome. Patients were maintained on nasal continuous positive airway pressure after each treatment. Patients received a mean 4.6 +/- 0.6 treatments for a mean total of 7915 +/- 1152 joules. There was no significant change in speech or swallowing at 3 months after completion of treatment. Patients reported a significant decrease in sleepiness with a mean change in ESS from 12.4 +/- 2.9 to 7.3 +/- 3.0 (P < 0.001). Mean apnea/hypopnea index decreased from 35.1 +/- 18.1 to 15.1 +/- 17.4 (P < 0.001). Transient mild to moderate pain and swelling occurred after each treatment. There were no significant complications (ulceration, paresthesia, infection). TCRF can successfully treat the OSAS patient with tongue base obstruction. Combined treatment of the ventral (genioglossus insertion) and dorsal tongue appears safe and may improve outcome with less total energy when compared with traditional dorsal-only applications.


Prevention of cerebral vasospasm by a humanized anti-CD11/CD18 monoclonal antibody administered after experimental subarachnoid hemorrhage in nonhuman primates.
Leukocyte-endothelial cell interactions occurring in the first hours after subarachnoid hemorrhage (SAH) initiate changes in the endothelium and vessel wall that lead to an influx of leukocytes and the development of chronic vasospasm days later. Upregulation of intercellular adhesion molecule-1 (ICAM-1), also called CD54, appears to be a crucial step in this process. There is increasing experimental evidence that blocking the interaction between ICAM-1, which is expressed on endothelium, and integrins such as lymphocyte function-associated antigen-1 (CD11a/CD18) and macrophage antigen-1 (complement receptor 3, CD11b/CD18), which are expressed on the surface of leukocytes,prevents not only inflammation of vessel walls but also chronic vasospasm. The authors extend their previous work with monoclonal antibody (mAb) blockade of leukocyte migration to a nonhuman primate model of chronic, posthemorrhagic cerebral vasospasm. Before surgery was performed, six young adult male cynomolgus monkeys underwent baseline selective biplane common carotid and vertebrobasilar artery cerebral angiography via a transfemoral route. On Day 0, a right frontosphenotemporal craniectomy was performed with arachnoid microdissection and placement of 2 to 3 ml of clotted autologous blood in the ipsilateral basal cisterns. The animals were given daily intravenous infusions of 2 mg/kg of either a humanized anti-CD11/CD18 or a placebo mAb beginning 30 to 60 minutes postoperatively. The monkeys were killed on Day 7 after a repeated selective cerebral angiogram was obtained. The area of contrast-containing vessels observed in each hemisphere on anteroposterior angiographic views was calculated for the angiograms obtained on Day 7 and expressed as a percentage of the area on baseline angiograms (percent control areal fraction). Review of flow cytometry and enzyme immunoassay data confirmed the presence of the anti-CD11/CD18 antibody in the serum and bound to leukocytes in the peripheral blood of treated animals. Comparisons of the groups revealed 53 +/- 4.8% control vascular areal fraction in the placebo group (two animals) and 95.8 +/- 9.4% in the anti-CD11/CD18-treated group (three animals), a statistically significant difference (p = 0.043, t-test). These results show that blockade of leukocyte migration into the subarachnoid space by an anti-CD11/CD18 mAb is effective in preventing experimental cerebral vasospasm in nonhuman primates, despite the unaltered presence of hemoglobin in the subarachnoid space. These experimental data support the hypothesis that inflammation plays a role in cerebral vasospasm after SAH.


Effectiveness, Safety, and Barriers to Early Mobilization in the Intensive Care Unit.
Patients admitted to the intensive care unit (ICU) are generally confined to bed leading to limited mobility that may have detrimental effects on different body systems. Early mobilization prevents or reduces these effects and improves outcomes in patients following critical illness. The purpose of this review is to summarize different aspects of early mobilization in intensive care. Electronic databases of PubMed, Google Scholar, ScienceDirect, and Scopus were searched using a combination of keywords. Full-text articles meeting the inclusion criteria were selected. Fifty-six studies on various aspects such as the effectiveness of early mobilization in various intensive care units, newer techniques in early mobilization, outcome measures for physical function in the intensive care unit, safety, and practice and barriers to early mobilization were included. Conclusion: Early mobilization is found to have positive effects on various outcomes in patients with or without mechanical ventilation. The newer techniques can be used to facilitate early mobilization. Scoring systems-specific to the ICU-are available and should be used to quantify patients' status at different intervals of time. Early mobilization is not commonly practiced in many countries. Various barriers to early mobilization have been identified, and different strategies can be used to overcome them.


Sympathetic nerve activity in the congenital long-QT syndrome.
Patients with congenital long-QT syndrome (LQTS) are susceptible to life-threatening arrhythmias. The sympathetic nervous system may have an important triggering role for cardiovascular events in LQTS. We therefore examined measurements of sympathetic neural traffic in patients with LQTS and matched control subjects. Twelve patients with congenital LQTS and 12 healthy volunteers matched for age, sex, and body mass index were studied. Heart rate, respiration, blood pressure, and sympathetic nerve activity to the skeletal muscle blood vessels (muscle sympathetic nerve activity) and to the skin (skin sympathetic nerve activity) were monitored and recorded continuously. Resting heart rate (P=0.03), muscle sympathetic nerve activity burst rate (P=0.008), and burst incidence (P=0.02) were lower in patients with LQTS than in control subjects. However, skin sympathetic nerve activity was very similar in patients with LQTS and control subjects. Spectral analysis of RR variability showed a decreased low-frequency component, an increased high-frequency component, and a decrease in the ratio of the low-frequency component to the high-frequency component in patients with LQTS (P=0.01). LQTS is associated with a selective reduction in sympathetic drive to muscle blood vessels and perhaps also to the heart.


Coronary heart disease and stroke disease burden attributable to fruit and vegetable intake in Japan: projected DALYS to 2060.
Fruit and vegetable consumption was considered a protective effect against cardiovascular and cerebrovascular diseases (CVDs). This study aimed to project the reduction in the CVD burden under different scenarios of increased fruit and vegetable intake in Japan by 2060. Population attributable fractions (PAF) were calculated by gender and age in 2015. The projection considered five scenarios for 2015, 2030, 2045, and 2060: 1) a baseline of no changes in intake; 2) a moderate increase in fruit intake (extra 50 g/day or 1/2 serving); 3) an high increase in fruit intake (extra 100 g/day or 1 serving); 4) a moderate increase in vegetable intake (extra 70 g/day or 1 serving); and 5) an high increase in vegetable intake (extra 140 g/day or 2 servings). Potentially preventable disability-adjusted life years (DALYs) for CVDs were estimated for each scenario. Monte Carlo simulations were performed to calculate the 95% confidence intervals of the estimates. Across all age groups, men had a higher daily vegetable intake than women (292.7 g/d > 279.3 g/d) but a lower daily fruit intake (99.3 g/d < 121.0 g/d). Comparing with recommended intake level (350 g/d of vegetable and 200 g/d of fruit), the total CVD burden was estimated to be 302,055 DALYs attributable to inadequate fruit consumption in 2015, which accounted for 12.6% of the total CVD burden (vegetable: 202,651 DALYs; 8.5%). In 2060, the percentage of the CVD burden due to insufficient intake of fruit is estimated to decrease to 7.9% under the moderate increase scenario and to decrease to 4.5% under the high increase scenario (vegetable: 5.4%; 2.4%). The study suggested that a relevantly large percentage of the CVD burden can be alleviated by promoting even modest increases in fruit and vegetable consumption in Japan.


1H NMR and UV-visible data fusion for determining Sudan dyes in culinary spices.
Two data fusion strategies (variable and decision level) combined with a multivariate classification approach (Partial Least Squares-Discriminant Analysis, PLS-DA) have been applied to get benefits from the synergistic effect of the information obtained from two spectroscopic techniques: UV-visible and (1)H NMR. Variable level data fusion consists of merging the spectra obtained from each spectroscopic technique in what is called "meta-spectrum" and then applying the classification technique. Decision level data fusion combines the results of individually applying the classification technique in each spectroscopic technique. Among the possible ways of combinations, we have used the fuzzy aggregation connective operators. This procedure has been applied to determine banned dyes (Sudan III and IV) in culinary spices. The results show that data fusion is an effective strategy since the classification results are better than the individual ones: between 80 and 100% for the individual techniques and between 97 and 100% with the two fusion strategies.


Peptide transport and animal growth: the fish paradigm.
Protein digestion products are transported from the intestinal lumen into the enterocyte both in the form of free amino acids (AAs), by a large variety of brush border membrane AA transporters, and in the form of di/tripeptides, by a single brush border membrane transporter known as PEPtide Transporter 1 (PEPT1). Recent data indicate that, at least in teleost fish, PEPT1 plays a significant role in animal growth by operating, at the gastrointestinal level, as part of an integrated response network to food availability that directly supports body weight. Notably, PEPT1 responds to both fasting and refeeding and is involved in a phenomenon known as compensatory growth (a phase of accelerated growth when food levels are restored after a period of growth depression). In particular, PEPT1 expression decreases during fasting and increases during refeeding, which is the opposite of what observed so far in mammals and birds. These findings in teleost fish document, to our knowledge, for the first time in a vertebrate model, a direct correlation between the expression of an intestinal transporter, such as PEPT1, primarily involved in the uptake of dietary protein degradation products and animal growth.


EP receptor-mediated inhibition by prostaglandin E(1) of cardiac L-type Ca(2+) current of rabbits.
Prostaglandin E(1) (PGE(1)) has cardioprotective effects on the ischemic-reperfused heart. To clarify the mechanisms underlying the protective action of PGE(1) on myocardium, we examined the effect of PGE(1) on the L-type Ca(2+) current (I(Ca)) using single atrial cells from rabbits. PGE(1) did not show a significant effect on basal I(Ca) but inhibited the I(Ca) prestimulated by isoproterenol (Iso, 30 nM). This inhibition was concentration dependent (EC(50) = 0.027 microM). Both sulprostone, a specific PGE receptor subtype (EP(1) and EP(3)) agonist, and 11-deoxy-PGE(1), an EP(3) agonist, inhibited the Iso-stimulated I(Ca), similar to PGE(1). Pretreatment with pertussis toxin (PTX) abolished the PGE(1) inhibition of I(Ca). Both the application of forskolin plus IBMX and intracellular dialysis with 8-bromoadenosine 3',5'-cyclic monophosphate eliminated the effect of PGE(1). PGE(1) did not show any further inhibition of I(Ca) when the effect of Iso was almost fully antagonized by acetylcholine. Methylene blue (guanylate cyclase inhibitor), KT-5823 (cGMP-dependent protein kinase inhibitor), and erythro-9-(2-hydroxy-3-nonyl)adenine (type II phosphodiesterase inhibitor) did not significantly change the inhibitory effect of PGE(1). These findings suggest that 1) PGE(1) inhibits Iso-stimulated I(Ca) by binding to the EP(3) receptor and 2) the PTX-sensitive and cAMP-dependent pathway is involved in the PGE(1) inhibition of I(Ca), but the nitric oxide-cGMP-dependent pathway is not. The PGE(1)-induced antiadrenergic effect shown in this study may contribute to the PGE(1) protection of myocardium against ischemia.


The preemptive analgesic effect of ketorolac and propacetamol for adenotonsillectomy in pediatric patients.
Both ketorolac and propacetamol are used postoperatively to control mild to moderate pain. This study compared the analgesic efficacy of ketorolac and propacetamol delivered either preoperatively or postoperatively, and assessed the preemptive analgesic effect of ketorolac and propacetamol for adenotonsillectomy. One hundred and two pediatric patients were divided randomly into four groups. The K1 and P1 groups received ketorolac 1 mg/kg or propacetamol 30 mg/kg after induction, respectively, whereas the K2 and P2 groups received each drug at the end of the operation, respectively. After adenotonsillectomy, we measured the NRS (Numerical Rating Scale), FPS (Faces Pain Scale) and OPS (Objective Pain scale) at 15, 30 and 60 min after arriving at the postanesthesia care unit. There were no significant differences in the NRS, FPS and OPS between K1 and K2 and between P1 and P2 for 60 min after operation at the postanesthesia care unit. These results suggest that both ketorolac (1 mg/kg) and propacetamol (30 mg/kg) have no preemptive analgesic effects during 1 hour after adenotonsillectomy.


Phenylalanine utilization by the gut and liver measured with intravenous and intragastric tracers in pigs.
To investigate intestinal and hepatic metabolism of phenylalanine, four conscious pigs (7.5 kg), bearing arterial, venous, and hepatic portal catheters, were fasted for 12 h and infused with [phenyl-2H5]phenylalanine via a peripheral vein and [carboxyl-13C]phenylalanine via the stomach. During the first 6 h of the infusion, the pigs remained fasted and received only the intravenous tracer. During the second 6 h, they received an intragastric infusion of milk replacer and both tracers. In the fasted state, the portal-drained viscera extracted 10% (P < 0.025) of the arterial [2H5]phenylalanine flow of the pigs. In the fed state, the splanchnic tissues metabolized 45% of the enteral tracer and intestinal metabolism accounted for 76% of the total splanchnic extraction. The tracer-to-tracee ratio of both tracers in apolipoprotein B-100 (apo B-100) phenylalanine was twofold (P < 0.001) higher than that of hepatic free phenylalanine. The ratios of the two tracers in portal (13C/2H; 1.66) and apo B-100 (1.76) phenylalanine were similar but higher (P < 0.05) than that of arterial phenylalanine (1.29). We conclude that intestinal metabolism dominates the splanchnic extraction of enteral phenylalanine and that in the fed state, the hepatic protein synthetic precursor pool derives from portal phenylalanine.


A highly specific antibody against the core fucose of the N-glycan in IgG identifies the pulmonary diseases and its regulation by CCL2.
Glycan structure is often modulated in disease or predisease states, suggesting that such changes might serve as biomarkers. Here, we generated a monoclonal antibody (mAb) against the core fucose of the N-glycan in human IgG. Notably, this mAb can be used in Western blotting and ELISA. ELISA using this mAb revealed a low level of the core fucose of the N-glycan in IgG, suggesting that the level of acore fucosylated (noncore fucosylated) IgG was increased in the sera of the patients with lung cancer, chronic obstructive pulmonary disease, and interstitial pneumonia compared to healthy subjects. In a coculture analysis using human lung adenocarcinoma A549 cells and antibody-secreting B cells, the downregulation of the FUT8 (α1,6 fucosyltransferase) gene and a low level of core fucose of the N-glycan in IgG in antibody-secreting B cells were observed after coculture. A dramatic alteration in gene expression profiles for cytokines, chemokines, and their receptors were also observed after coculturing, and we found that the identified C-C motif chemokine 2 was partially involved in the downregulation of the FUT8 gene and the low level of core fucose of the N-glycan in IgG in antibody-secreting B cells. We also developed a latex turbidimetric immunoassay using this mAb. These results suggest that communication with C-C motif chemokine 2 between lung cells and antibody-secreting B cells downregulate the level of core fucose of the N-glycan in IgG, i.e., the increased level of acore fucosylated (noncore fucosylated) IgG, which would be a novel biomarker for the diagnosis of patients with pulmonary diseases.


Insulin-like growth factor-1 regulates platelet activation through PI3-Kalpha isoform.
Platelets release insulin-like growth factor-1 (IGF-1) from alpha granules upon activation. We have investigated the regulation of IGF-1 in G(i)-dependent pathways leading to Akt activation and the role of IGF-1 in platelet activation. IGF-1 alone failed to induce platelet aggregation, but IGF-1 potentiated 2-MeSADP-induced platelet aggregation in a concentration-dependent manner. IGF-1 triggered platelet aggregation in combination with selective P2Y(1) receptor activation. IGF-1 also caused platelet aggregation without shape change when combined with selective G(z) stimulation by epinephrine, suggesting the role of IGF-1 in platelet aggregation by supplementing G(i) pathways. The potentiating effect of IGF-1 was not affected by intracellular calcium chelation. Importantly, IGF-1 was unable to potentiate platelet aggregation by the phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin, suggesting a critical regulation by PI3-K. Moreover, the potentiating effect of IGF-1 was abolished by the presence of PI3-K p110alpha inhibitor PIK-75. Stimulation of platelets with IGF-1 resulted in phosphorylation of Akt, a downstream effector of PI3-K, which was completely inhibited by wortmannin. IGF-1-induced Akt phosphorylation was abolished by PIK-75 suggesting the contribution of PI3-K p110alpha for activation of Akt by IGF-1. These results demonstrate that IGF-1 plays a role in potentiating platelet aggregation by complementing G(i)- but not G(q)-signaling pathways via PI3-K p110alpha.


The Effect of a Mechanical Compression Device and Supraglottic Airway on Flow Time: A Simulation Study of Out-of-Hospital Cardiac Arrest in a High-Rise Building.
High-rise buildings present unique challenges to providing high-quality CPR. We investigated the effect of using a mechanical compressor and supraglottic airway on flow time and CPR quality in simulated cardiac arrests occurring within a high-rise building. Twelve teams of EMS providers performed CPR according to 4 scenarios: manual compression and ventilation through bag-valve-mask (MAB) or supraglottic airway (MAS); mechanical compression and ventilation through bag-valve-mask (MEB) or supraglottic airway (MES). Chest compression indices did not differ significantly among the groups. The mechanical compression groups had a higher flow time fraction from exiting the elevator until the manikin was loaded into the ambulance than the manual compression groups. The supraglottic airway groups had higher flow time fractions from entering the elevator until the end of the scenario than the bag-valve-mask groups. The total flow time fraction was lowest in the MAB group and was highest in the MEB group (P < 0.001). In simulated cardiac arrest in a high-rise building, the use of a supraglottic airway maintained flow time at a level similar to that observed with the use of a mechanical compressor. Moreover, the use of a mechanical compressor and a supraglottic airway increased the flow time most effectively.


Association between boarding in the emergency department and in-hospital mortality: A systematic review.
Boarding in the emergency department (ED) is a critical indicator of quality of care for hospitals. It is defined as the time between the admission decision and departure from the ED. As a result of boarding, patients stay in the ED until inpatient beds are available; moreover, boarding is associated with various adverse events. The objective of our systematic review was to determine whether ED boarding (EDB) time is associated with in-hospital mortality (IHM). A systematic search was conducted in academic databases to identify relevant studies. Medline, PubMed, Scopus, Embase, Cochrane, Web of Science, Cochrane, CINAHL and PsychInfo were searched. We included all peer-reviewed published studies from all previous years until November 2018. Studies performed in the ED and focused on the association between EDB and IHM as the primary objective were included. Extracted data included study characteristics, prognostic factors, outcomes, and IHM. A search update in PubMed was performed in May 2019 to ensure the inclusion of recent studies before publishing. From the initial 4,321 references found through the systematic search, the manual screening of reference lists and the updated search in PubMed, a total of 12 studies were identified as eligible for a descriptive analysis. Overall, six studies found an association between EDB and IHM, while five studies showed no association. The last remaining study included both ICU and non-ICU subgroups and showed conflicting results, with a positive association for non-ICU patients but no association for ICU patients. Overall, a tendency toward an association between EDB and IHM using the pool random effect was observed. Our systematic review did not find a strong evidence for the association between ED boarding and IHM but there is a tendency toward this association. Further well-controlled, international multicenter studies are needed to demonstrate whether this association exists and whether there is a specific EDB time cut-off that results in increased IHM.


Facets on the psychopathy checklist screening version and instrumental violence in forensic psychiatric patients.
There is a recognised relationship between psychopathy and instrumental violence, but not all violence by people who meet the criteria for psychopathy is instrumental. Our aims were to compare offence types among forensic psychiatric patients with and without the Psychopathy Checklist: Screening Version (PCL: SV) criteria for psychopathy. Our specific questions were whether factor 1 - the interpersonal affective dimension - was related to instrumentality and on severity of the violent crime. Our hypothesis was that the relationship between psychopathy and instrumental violence would be dependent on the severity of the violent crime. Sixty-five male patients at the forensic psychiatric hospital in Sundsvall, all with a violent criminal history, were assessed for psychopathy through interview and records using the PCL: SV. Severity and the instrumentality of their previous violence were coded using the Cornell coding guide for violent incidents. The interpersonal features of psychopathy (the interpersonal facet), and only the interpersonal features were significantly associated with instrumentality and severity of violence. Instrumentality was also significantly related to the severity of the violence, independent of psychopathy score. The results indicated that, at least among forensic psychiatric patients, planning is more likely than not with respect to serious crimes. The specific link between interpersonal features of psychopathy and instrumental and severe violence suggests potential clinical value in recognising subtypes of psychopathy.


Pharmacokinetics of dexmedetomidine in anaesthetized horses following repeated subcutaneous administration and intravenous constant rate infusion.
The inclusion of dexmedetomidine (DEX) within a balanced general anaesthesia protocol is effective in improving the clinical outcome and recovery quality of anaesthesia in horses. This study aimed to determine the pharmacokinetic profile of DEX following repeated subcutaneous (SC) administration at 2 µg/kg every 60 min till the end of the procedure in comparison to intravenous constant rate infusion (CRI) at 1 µg/kg/h in anaesthetized horses undergoing diagnostic procedures up to the end of the diagnostic procedure. In the CRI and SC groups DEX maximum concentrations (Cmax) were 0.83 ± 0.27 ng/mL and 1.14 ± 0.71 ng/mL, respectively, reached at a time (Tmax) of 57.0 ± 13.4 min and 105.5 ± 29.9 min. Mean residence time to the last measurable concentration (MRTlast) was 11.7 ± 6.2 and 55.8 ± 19.7 min for the CRI group and SC groups, respectively. The apparent elimination half-life was 18.0 ± 10.0 min in the CRI group and 94.8 ± 69.8 min for the SC group, whereas the area under the curve (AUC0-last) resulted 67.7 ± 29.3 and 83.2 ± 60.5 min*ng/mL for CRI and SC group, respectively. Clearance was 16.26 ± 8.07 mL/min/kg for the CRI group. No signs of adverse effects were recorded in both groups. The pharmacokinetic profile of DEX following repeated SC administration in anaesthetized horses was comparable to intravenous CRI administration during the intranaesthetic period and beneficial during the recovery phase from general anaesthesia. The SC route could be considered as an alternative to CRI for improving the recovery quality of equine patients undergoing general anaesthesia.


An Exploratory Study on Exemplary Practice of Nurse Consultants.
To examine the exemplary practice of nurse consultants (NCs) and derive a model to illustrate the highest level of advanced nursing practice. A descriptive study was conducted to examine the practice and outcomes of seven NC roles in varied clinical specialties in Hong Kong. Exemplary practice was examined in relation to competencies for advanced practice nursing in Hong Kong and the United Kingdom. Data about NC characteristics and their practices were collected using a structured questionnaire and analyzed using descriptive statistics. Health service documents and clinical notes were analyzed using the framework approach. All NCs demonstrated the competence expected of an advanced practice nurse with impacts on patients, nursing profession, and the organization as identified in the advanced nursing practice framework in Hong Kong. NCs also performed at the highest level of practice delineated by Skills for Health in the United Kingdom. They were involved in diagnostic and therapeutic practice, and identified patient satisfaction and symptom management as key outcomes. This study provides new insight into levels of advanced practice and illustrates the exemplary work of NCs to demonstrate how they have developed and shaped services to bring about positive patient and organizational outcomes. Career laddering that places NCs at the highest level of advanced practice is important for making the best use of nursing expertise to achieve optimal patient and organizational outcomes. This study addresses a knowledge gap to enrich our current understanding of the impact of advanced practice nursing roles by linking NC role practices and competencies to key outcomes.


Chromosome aberrations evaluated by comparative genomic hybridization in B-cell chronic lymphocytic leukemia: correlation with CD38 expression.
B-cell chronic lymphocytic leukemia (B-CLL) results from the accumulation of monoclonal CD5+ B cells. Despite its homogeneity at cellular level, B-CLL is clinically heterogeneous. Clinical studies indicate that CD38+ B-CLL are characterized by a more aggressive clinical course than are CD38- B-CLL. On the basis of these studies and considering the established correlation between specific chromosome aberrations and the clinical course of B-CLL, it is possible that CD38+ B-CLL cases are also characterized by specific subsets of chromosomal alterations. Comparative genomic hybridization (CGH) was performed on purified B-cells from peripheral blood of 52 patients with B-CLL in order to detect chromosome imbalance. The immunophenotype of the patients, including CD38 expression, was also determined by flow cytometry. The results of CGH experiments were then compared with CD38 expression. We found a clear correlation between the presence of chromosomal imbalances and CD38 expression: 13/16 CD38+ cases had chromosome imbalances, most of them (12/13) correlated with a poor prognosis. Among the CD38- B-CLL patients, only 8/36 displayed chromosome imbalances; the only three cases with loss in 13q as a single aberration, considered a good prognostic marker, were in this group. Moreover, we found that cytogenetic alterations were also more complex in the CD38+ B-CLL subset, since 9/10 with two or more aberrations were in the CD38+ group. Collectively, the data reinforce the value of CD38 as a prognostic factor and indicate that genotypic/phenotypic features distinguish B-CLL subsets.


Bcl-2 immunohistochemistry in a surgical series of non-small cell lung cancer patients.
The bcl-2 gene is implicated in oncogenesis by its ability to prolong cell survival through the inhibition of apoptosis, without increasing cell proliferation. An association between immunohistochemical staining for bcl-2 protein and the histological type and prognosis of non-small cell carcinoma was hypothesized by Pezzella et al. (N Engl J Med 329:690-694, 1993). In a case series, we stained formalin-fixed, paraffin-embedded tumor tissue from 106 surgical non-small cell lung cancer patients with an antibody to bcl-2 protein (DAKO clone 124, Carpinteria, CA). The resulting bcl-2 staining data were evaluated for associations with demographic, histological, immunohistochemical, and genetic features, including p53 mutations. Bcl-2 staining was observed in tumors from 29 of 106 (27%) of subjects, but was significantly less frequent in subjects' adenocarcinoma histology (8 of 55, 14.6%) (P = .007). This finding persisted after adjustment for age, gender, stage, grade, smoking history, and disease-free survival. In univariate analyses, no association was seen with age, weight, body mass index, gender, or pack-years smoking; tumor grade, stage, or patient performance status; p53 or c-erbB2 immunohistochemical staining, or p53 mutations. These data agree with earlier reports that bcl-2 staining is less common in adenocarcinomas; however, our data do not support the hypothesis that bcl-2 staining confers a better prognosis overall, in squamous cell carcinoma, or in an older patient population.


Treating Perioperative and Acute Pain in Patients on Buprenorphine: Narrative Literature Review and Practice Recommendations.
Opioid use disorder (OUD), a leading cause of morbidity and mortality in the USA, can be effectively treated with buprenorphine. However, the same pharmacologic properties (e.g., high affinity, partial agonism, long half-life) that make it ideal as a treatment for OUD often cause concern among clinicians that buprenorphine will prevent effective management of acute pain with full agonist opioid analgesics. Because of this concern, many patients are asked to stop buprenorphine preoperatively or at the onset of acute pain, placing them at high risk for both relapse and a difficult transition back to buprenorphine after acute pain has resolved. The purpose of this review is to summarize the existing literature for acute pain and perioperative management in patients treated with buprenorphine for OUD and to provide practical management recommendations for generalist practitioners based on evidence and clinical experience. In short, evidence suggests that sufficient analgesia can be achieved with maintenance of buprenorphine and use of both opioid and non-opioid analgesic options for breakthrough pain. We recommend that clinicians (1) continue buprenorphine in the perioperative or acute pain period for patients with OUD; (2) use a multi-modal analgesic approach; (3) pay attention to care coordination and discharge planning when making an analgesic plan for patients with OUD treated with buprenorphine; and (4) use an individualized approach founded upon shared decision-making. Clinical examples involving mild and severe pain are discussed to highlight important management principles.


Restoration of basic hand function by double transpositional digital replantation in five-digit amputations.
In multiple digital amputations with severe crush injury, replantation of digits with a poor prognosis is often fruitless. Those digits with a favorable prognosis should be selectively replanted in positions that provide optimal functional outcome, regardless of their anatomic origins: this is the principle of transpositional replantation. The authors present a case of right-hand crush injury with amputation of all five digits. Basic hand function was restored by double transpositional digital replantations without additional reconstructive procedures. In properly selected cases, such procedures can preserve greater hand function and reduce the necessity for secondary operations. It is suggested that transpositional replantation should be considered when multiple digital amputations are encountered.


Cultural innovations and demographic change.
Demography plays a large role in cultural evolution through its effects on the effective rate of innovation. If we assume that useful inventions are rare, then small isolated societies will have low rates of invention. In small populations, complex technology will tend to be lost as a result of random loss or incomplete transmission (the Tasmanian effect). Large populations have more inventors and are more resistant to loss by chance. If human populations can grow freely, then a population-technology-population positive feedback should occur such that human societies reach a stable growth path on which the rate of growth of technology is limited by the rate of invention. This scenario fits the Holocene to a first approximation, but the late Pleistocene is a great puzzle. Large-brained hominins existed in Africa and west Eurasia for perhaps 150,000 years with, at best, slow rates of technical innovation. The most sophisticated societies of the last glacial period appear after 50,000 years ago and were apparently restricted to west and north-central Eurasia and North Africa. These patterns have no simple, commonly accepted explanation. We argue that increased high-frequency climate change around 70,000-50,000 years ago may have tipped the balance between humans and their competitor-predators, such as lions and wolves, in favor of humans. At the same time, technically sophisticated hunters would tend to overharvest their prey. Perhaps the ephemeral appearance of complex tools and symbolic artifacts in Africa after 100,000 years ago resulted from hunting inventions that allowed human populations to expand temporarily before prey overexploitation led to human population and technology collapse. Sustained human populations of moderate size using distinctively advanced Upper Paleolithic artifacts may have existed in west Eurasia because cold, continental northeastern Eurasia-Beringia acted as a protected reserve for prey populations.


Gastrin, somatostatin, G and D cells of gastric ulcer in rats.
To investigate the relationship among gastrin, somatostatin, G and D cells in gastric ulcer and in its healing process in rats. Fourty-nine Wistar rats were divided into 7 groups. The gastric ulcer model was induced by acetic acid successfully. The gastrin and the somatostatin in rat plasma, gastric fluid and antral tissue were measured by radioimmunoassay(RIA). G and D cells in antral mucosa were analyzed with polyclonal antibody of gastrin and somatostatin by immunohistochemical method and Quantimet 500 image analysis system. In gastric ulcer, the level of gastrin in plasma, gastric fluid, and antral tissue increased, that of somatostatin declined, and the disorder gradually recovered to the normal level in the healing process. Immunohistochemical technique of G and D cells in antral mucosa demonstrated that the number of G cells increased and that of D cells decreased, both areas of G and D cells declined, the ratio of number and area of G/D increased in gastric ulcer, and the disorder gradually recovered in the healing process. In gastric ulcer, the increased gastrin secreted by G cells, the declined somatostatin secreted by D cells, and the disordered G/D cell ratio can lead to gastrointestinal dysfunction.


Implementation of a Wearable Ultrasound Device for the Overnight Monitoring of Tongue Base Deformation during Obstructive Sleep Apnea Events.
Obstructive sleep apnea (OSA) is a breathing disorder characterized by the repeated collapse of the pharyngeal airway during sleep. Previous studies have reported that tongue base deformation may be a major contributing factor. However, overnight monitoring of tongue motion in patients with OSA has previously been impracticable. We developed a wearable ultrasound device for prolonged recording during natural sleep of the changes in tongue base thickness (TBT) in patients with OSA. The maximum TBT was fed into a polysomnography system so that physiologic signals and TBT data were simultaneously monitored. Subject trials revealed that TBT increased significantly during snoring, hypopnea and apnea events during natural sleep in patients with OSA. Moreover, the data revealed that the location of the maximum TBT during normal breathing was significantly different compared with the location during obstructive respiratory events, which implies a posterior or inferior displacement of the tongue base during sleep apnea.


Real-space observation of far- and near-field-induced photolysis of molecular oxygen on an Ag(110) surface by visible light.
Dissociation of molecular oxygen is an important elementary process in heterogeneous catalysis. Here, we report on a real-space observation of oxygen photolysis on the Ag(110) surface at 78 K by far- and near-field excitation in the ultraviolet-near-infrared range using a low-temperature scanning tunneling microscope (STM) combined with wavelength-tunable laser excitation. The photolysis of isolated oxygen molecules on the surface occurs even by visible light with the cross section of ∼10-19 cm2. Time-dependent density functional theory calculations reveal optical absorption of the hybridized O2-Ag(110) complex in the visible and the near-infrared range which is associated with the oxygen photolysis. We suggest that the photolysis mechanism involves a direct charge transfer process. We also demonstrate that the photolysis can be largely enhanced in plasmonic STM junctions, and the cross section is estimated to be ∼10-17 cm-2 in the visible and the near-infrared range, which appears to be an interesting feature of plasmon-induced reactions from the perspective of photochemical conversion with the aid of solar energy.


Adaptive optics stochastic optical reconstruction microscopy (AO-STORM) using a genetic algorithm.
The resolution of Single Molecule Localization Microscopy (SML) is dependent on the width of the Point Spread Function (PSF) and the number of photons collected. However, biological samples tend to degrade the shape of the PSF due to the heterogeneity of the index of refraction. In addition, there are aberrations caused by imperfections in the optical components and alignment, and the refractive index mismatch between the coverslip and the sample, all of which directly reduce the accuracy of SML. Adaptive Optics (AO) can play a critical role in compensating for aberrations in order to increase the resolution. However the stochastic nature of single molecule emission presents a challenge for wavefront optimization because the large fluctuations in photon emission do not permit many traditional optimization techniques to be used. Here we present an approach that optimizes the wavefront during SML acquisition by combining an intensity independent merit function with a Genetic algorithm (GA) to optimize the PSF despite the fluctuating intensity. We demonstrate the use of AO with GA in tissue culture cells and through ~50µm of tissue in the Drosophila Central Nervous System (CNS) to achieve a 4-fold increase in the localization precision.


Replication bypass of the acrolein-mediated deoxyguanine DNA-peptide cross-links by DNA polymerases of the DinB family.
DNA-protein cross-links (adducts) are formed in cellular DNA under a variety of conditions, particularly following exposure to an alpha,beta-unsaturated aldehyde, acrolein. DNA-protein cross-links are subject to repair or damage-tolerance processes. These adducts serve as substrates for proteolytic degradation, yielding DNA-peptide lesions that have been shown to be actively repaired by the nucleotide excision repair complex. Alternatively, DNA-peptide cross-links can be subjected to replication bypass. We present new evidence about the capabilities of DNA polymerases to synthesize DNA past such cross-links. DNAs were constructed with site-specific cross-links, in which either a tetrapeptide or a dodecylpeptide was covalently attached at the N (2) position of guanine via an acrolein adduct, and replication bypass assays were carried out with members of the DinB family of polymerases, human polymerase (pol) kappa, Escherichia coli pol IV, and various E. coli polymerases that do not belong to the DinB family. Pol kappa was able to catalyze both the incorporation and the extension steps with an efficiency that was qualitatively indistinguishable from control (undamaged) substrates. Fidelity was comparable on all of these substrates, suggesting that pol kappa would have a role in the low mutation frequency associated with replication of these adducts in mammalian cells. When the E. coli orthologue of pol kappa, damage-inducible DNA polymerase, pol IV, was analyzed on the same substrates, pause sites were detected opposite and three nucleotides beyond the site of the lesion, with incorporation opposite the lesion being accurate. In contrast, neither E. coli replicative polymerase, pol III, nor E. coli damage-inducible polymerases, pol II and pol V, could efficiently incorporate a nucleotide opposite the DNA-peptide cross-links. Consistent with a role for pol IV in tolerance of these lesions, the replication efficiency of DNAs containing DNA-peptide cross-links was greatly reduced in pol IV-deficient cells. Collectively, these data indicate an important role for the DinB family of polymerases in tolerance mechanisms of N (2)-guanine-linked DNA-peptide cross-links.


The tunnel sponge--an explant for retinal detachment surgery.
The tunnel sponge explant for retinal detachment surgery is a modification of the cylindrical, silastic sponge. A central tunnel exists along the length of the sponge allowing a silicone band to be threaded internally. A high, localized indentation can be achieved as well as encirclement with minimal volume displacement of the globe.


Antibody alone does not prevent experimental cytomegalovirus retinitis in mice with retrovirus-induced immunodeficiency (MAIDS).
Passive-transfer studies were performed to assess the ability of antibody alone to reduce the frequency and/or severity of necrotizing retinitis caused by murine cytomegalovirus (MCMV) in C57BL/6 mice with retrovirus-induced immunodeficiency syndrome (MAIDS). Initial experiments showed a gradual decline in the ability of mice to initiate humoral immunity during the evolution of MAIDS so that neither MCMV-specific IgM nor IgG could be detected during late-stage MAIDS. Passively administered hyperimmune MCMV immunoglobulin, however, could be detected within the serum of mice with MAIDS for at least 9 days after intraperitoneal injection and protected these animals in preliminary experiments from systemic MCMV disease and death when administered 24 h prior to intraperitoneal challenge with a lethal dose of virus. Nonetheless, passive transfer of hyperimmune MCMV serum to mice with MAIDS failed to reduce intraocular MCMV titers, frequency of retinitis, or severity of retinitis when administered 24 h prior to subretinal MCMV inoculation. Whereas whole eyes of MAIDS animals that received normal mouse serum and were injected subretinally with MCMV had an ocular MCMV titer of 4.3 log10 and a frequency of retinitis of 89% (severity score = 55%), whole eyes of antibody-treated mice with MAIDS had an ocular MCMV titer of 4.3 log10 and a frequency of retinitis of 87% (severity score = 57 %). Passive transfer of a neutralizing MCMV-specific monoclonal antibody also failed to reduce the frequency or severity of MCMV retinitis when administered to mice with MAIDS prior to subretinal MCMV inoculation. Our findings suggest that antibody immunotherapy alone will not be effective therapeutically for cytomegalovirus retinitis in patients with AIDS.


Investigation on the infection mechanism of the fungus Clonostachys rosea against nematodes using the green fluorescent protein.
The fungus Clonostachys rosea (syn. Gliocladium roseum) is a potential biocontrol agent. It can suppress the sporulation of the plant pathogenic fungus Botrytis cinerea and kill pathogenic nematodes, but the process of nematode pathogenesis is poorly understood. To help understand the underlying mechanism, we constructed recombinant strains containing a plasmid with both the enhanced green fluorescent protein gene egfp and the hygromycin resistance gene hph. Expression of the green fluorescent protein (GFP) was monitored using fluorescence microscopy. Our observations reveal that the pathogenesis started from the adherence of conidia to nematode cuticle for germination, followed by the penetration of germ tubes into the nematode body and subsequent death and degradation of the nematodes. These are the first findings on the infection process of the fungal pathogen marked with GFP, and the developed method can become an important tool for studying the molecular mechanisms of nematode infection by C. rosea.


The Minnesota Multiphasic Personality Inventory-2 Restructured Form in National Guard soldiers screening positive for posttraumatic stress disorder and mild traumatic brain injury.
The Minnesota Multiphasic Personality Inventory-2 Restructured Form (MMPI-2 RF) was administered to 251 National Guard soldiers who had recently returned from deployment to Iraq. Soldiers were also administered questionnaires to identify posttraumatic stress disorder (PTSD) and mild traumatic brain injury (mTBI). On the basis of responses to the screening instruments, the National Guard soldiers who produced a valid MMPI-2 RF were classified into four groups: 21 soldiers who screened positive for PTSD only, 33 soldiers who screened positive for mTBI only, 9 soldiers who screened positive for both conditions, and 166 soldiers who did not screen positive for either condition. Results showed that the MMPI-2 RF was able to differentiate across the groups with the MMPI-2 RF specific problem scale Anxiety adding incrementally to MMPI-2 Restructured Clinical scales in predicting PTSD. Both MMPI-2 RC1 (Somatic Complaints) and MMPI-2 RF head pain complaints predicted mTBI screen but did not add incrementally to each other. Of note, all of the MMPI-2 RF validity scales associated with overreporting, including Symptom Validity-Revised (FBS-r), were not significantly elevated in the mTBI group. These findings support the use of the MMPI-2 RF in assessing PTSD in non-treatment-seeking veterans. This further suggests that a positive screen for mTBI alone is not associated with significant emotional disturbance.


Cellular pH and Na+-H+ exchange activity in lens epithelium of Bufo marinus toad.
Incubation of lenses with the acetoxymethyl ester of 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF-AM) led to selective, prolonged entrapment of the deesterified, pH-sensitive, membrane-impermeable form of the dye (BCECF) in the lens epithelial cells allowing, thereby, for the continuous fluorometric monitoring of epithelial intracellular pH (pHi) in intact lenses. Under anaerobic conditions in physiological solution, the epithelium maintained a pHi of 7.36 +/- 0.14 (mean +/- SD, n = 6 lenses). In the absence of CO2 and HCO3-, the pHi increased to 7.83 +/- 0.38 (n = 55). In this condition, either the removal of Na+ or the addition of amiloride to the medium resulted in a decrease of pHi. This acidification was increased in amiloride-inhibitable fashion when lenses were loaded with Na+ before extracellular Na+ removal. The extent of artificially generated cellular acidosis (induced by the "NH4+ loading" technique) was markedly decreased by the presence of extracellular Na+ or reverted on its addition when the ion was initially absent; the latter reversal was inhibited by amiloride. This inhibition was by-passed by monensin addition. The existence of a Na+-H+ antiport in the epithelial cells is concluded.


[Structural and functional organisation of the spleen lymphoid tissue after exposure to hypergravitation].
Using the stereologic methods and quantitative analysis of the cells of lymphoid series in the splenic white pulp of Wistar rats, the changes were studied after continuous 5-day-long exposure to hypergravitation of 2 G. The injurious effect of hypergravitation was manifested in the reduction of the dimensions of periarterial lymphoid sheaths (PALS) and of lymphoid nodules, marginal zone dilatation and the disappearance of lymphoid nodules with germinal centers. Changes in the cell population of the white pulp are indicative of the suppression of blast-transformation and cell reproductive activity (as demonstrated by the disappearance of blast cells and the mitotic cells) and the reduction of lymphocytopoiesis in the organ lymphoid tissue. The proportion of immature plasma cells is increased. The changes demonstrated are more pronounced in PALS than in lymphoid nodules. The significant cellular reorganization of the marginal zones is the reflection of compensatory processes, which develop in response to the injurious effect of hypergravitation on the lymphoid tissue of the spleen.


[Human ectoparasitoses: current status in Uruguay].
The present status of human ectoparasitoses in Uruguay is analyzed with the main purpose of performing in the future, comparative studies in the region. After classifying ectoparasites in temporary and permanent, comments are made on particular clinical and epidemiological aspects of some of them. For remarking: the high prevalence of pediculosis capitis and scabies; the frequent observation of cutaneous myiasis by Dermatobia hominis with a very interesting modification of the geographical national distribution of the fly and the presence of an endemic area of creeping eruption by Ancylostoma braziliense and A. caninum in the north of the country.


Identification of the type I trimethoprim-resistant dihydrofolate reductase specified by the Escherichia coli R-plasmid R483: comparison with procaryotic and eucaryotic dihydrofolate reductases.
We have isolated and determined the nucleotide sequence of a 1,626-base-pair fragment from R-plasmid R483 which encodes a trimethoprim-resistant dihydrofolate reductase. Analysis of the nucleotide sequence of this fragment revealed the presence of two open reading frames, each sufficient to encode polypeptides of approximately 17,000 daltons. Both open regions are preceded by sequences conforming closely to the canonical description of procaryotic promoters. A 490-base-pair HpaI fragment spanning one of the potential coding regions was inserted into a plasmid vector under the transcriptional control of the trp promoter. Cells transformed with this plasmid were trimethoprim resistant and produced dihydrofolate reductase activity which in vitro was resistant to moderate levels of trimethoprim. Analysis of the predicted amino acid sequence of this protein indicated that the R483-encoded trimethoprim-resistant enzyme was distantly related to the trimethoprim-sensitive bacterial homologs. The conserved amino acids were localized primarily to the region of the enzyme previously shown to comprise the hydrophobic substrate binding pocket.


Genetic disposition to deafness in maternal rubella: fact or myth?
Eleven parental pairs, each having a child deaf from maternal rubella, were evaluated with a battery of specialized audiologic tests. The tests which included measurement of Békésy hearing threshold and the stapedial reflex threshold at 0.5, 1.0, and 2.0 kHz were used to test the theory that these parents have a predisposition to elevated reflex thresholds, Békésy hearing threshold dips, and a history of familial deafness. The present study failed to find any history of familial deafness or Békésy dips. Elevated reflex thresholds were noted in seven couples. This proportion was shown to be significant (p less than 0.03). The relevance of these findings in light of current clinical practice is discussed. A comparison of pure tone hearing levels of the children of parents with and without elevated reflex thresholds, failed to show a significant difference.


Blockade of Ca2+ and K+ currents in bag cell neurons of Aplysia californica by dihydropyridine Ca2+ antagonists.
The effects of dihydropyridine calcium antagonists on whole-cell Ca2+ and K+ currents in the neurosecretory bag cells of the marine mollusc Aplysia californica have been investigated. Nifedipine and nisoldipine blocked bag cell Ca2+ currents with effects similar to those seen previously on Ca2+ currents in cardiac muscle: both compounds appeared to interact with Ca2+ channels when they were closed, open, and inactivated. Also, as seen in cardiac cells, nifedipine apparently binds with higher affinity to Ca2+ channels when they are inactivated than when they are either closed or open. Nifedipine and nisoldipine also inhibited 2 outward K+ currents in bag cells: the "delayed rectifier" (IK) and the "A" (IA) currents. Half-maximal blockade of Ca2+ currents occurred at approximately 1.4 microM nifedipine, compared to approximately 3-5 microM for half-maximal blockade of IK and IA. The effects of these compounds on bag cell Ca2+ and K+ currents are interpreted and discussed here in terms of the "modulated receptor" model of drug action. In contrast, however, no measurable effects of nifedipine or nisoldipine were seen on Ca2+ (and/or K+) currents in several vertebrate neuronal cell types. Our results suggest that there are likely to be structural and/or conformational variations in Ca2+ channels in different cells, tissues, and/or species and also that, in some cells, Ca2+ and K+ channels might be structurally similar. These findings also suggest, therefore, that if dihydropyridine binding is used to identify Ca2+ channels, care should be taken to ensure that binding correlates closely with the Ca2+ channels of interest.


The Relationship between Resource Utilization, Clinical Risk and Hospital Costs for Emergency Medical Admissions.
There has been little study of the relationship between resource utilisation, clinical risks and hospital costs in acute medicine with the question remaining as to whether current funding models reflect patient acuity. We examined the relationship between resource use for investigations/allied professional and patient episode costs in all emergency medical admissions admitted to our institution during 2008-2013. Univariate estimates were compared with a multivariate model adjusted for major cost predictors. Interestingly, the model adjusted cost estimates changed considerably when compared with univariate analysis. We used both linear and non-linear (quantile regression) methods due to the highly skewed nature of hospital costs. The data suggested that hospital episode costs were predictable and driven by objective measures of clinical complexity. The use of expensive investigations and healthcare professional time was secondary to the clinical acuity. Thus, cost was heavily weighted towards higher complexity, and lower resource utilisation associated with lower risk patient groups. However, the non-linear nature of the costings would caution against simple predictor models and non-linear techniques such as quantile regression may, as we have demonstrated, prove superior in defining the underlying relationships.


[The power of nursing versus minimal nursing data].
This article reports the Belgian experience of the Nursing Minimum Data Set (NMDS), in order to make the nurse work visible. The collected data are forwarded to the Health Ministry and, together with the DRGs, constitute the base for the determination of hospitals financing budget in Belgium.


Proposing a standardized protocol for raw biosignal transmission.
In this paper, we propose a standardized interface called TiA (TOBI interface A) to transmit raw biosignals, supporting multirate and block-oriented transmission of different kinds of signals from various acquisition devices (e.g., EEG, electrooculogram, near-infrared spectroscopy signals, etc.) at the same time. To facilitate a distinction between those kinds of signals, so-called signal types are introduced. TiA is a single-server, multiple-client system, whereby clients can connect to the server at runtime. Information transfer between client and server is divided into control and data connections. The control connections use transmission control protocol (TCP) and transmit extensible-markup-language (XML)-encoded meta information. The data transmission utilizes a user datagram protocol (UDP) or TCP with a binary data stream. A standardized handshaking procedure for the connection setup and a standardized binary data packet has been defined. Thus, a standardized layer, abstracting used hardware devices and facilitating distributed raw data transmission in a standardized way, has been evolved. A cross-platform library, implemented in C ++, is available for download.


Survey sampling issues in primary care research.
Survey research methods are often used to answer questions in primary care medicine. Consequently, investigators must understand and address sampling issues. Probability sampling designs have particular utility for questions that seem most appropriate for survey research. These designs provide each member of the population with a known probability of selection, and if performed properly with an appropriate sample size, they provide a sample with essentially no selection bias. Examples of random and stratified sampling designs are presented. These two probability designs, along with nonprobability sampling techniques (convenience, purposive, quota), are discussed in terms of bias, error, and possible limitations.


Sonic Hedgehog signaling is a positive oligodendrocyte regulator during demyelination.
The morphogen Sonic Hedgehog (Shh) controls the generation of oligodendrocyte (OLs) during embryonic development and regulates OL production in adulthood in the cortex and corpus callosum. The roles of Shh in CNS repair following lesions associated with demyelinating diseases are still unresolved. Here, we address this issue by using a model of focal demyelination induced by lysolecithin in the corpus callosum of adult mice. Shh transcripts and protein were not detected in control animals but were upregulated in a time-dependent manner in the oligodendroglial lineage within the lesion. We report an increased transcription of Shh target genes suggesting a broad reactivation of the Shh pathway. We show that the adenovirus-mediated transfer of Shh into the lesioned brain results in the attenuation of the lesion extent with the increase of OL progenitor cells (OPCs) and mature myelinating OL numbers due to survival, proliferation, and differentiation activities as well as the decrease of astrogliosis and macrophage infiltration. Furthermore, the blocking of Shh signaling during the lesion, using its physiological antagonist, Hedgehog interacting protein, results in a decrease of OPC proliferation and differentiation, preventing repair. Together, our findings identify Shh as a necessary factor playing a positive role during demyelination and indicate that its signaling activation stands as a potential therapeutic approach for myelin diseases.


The most heat-resistant conidia observed to date are formed by distinct strains of Paecilomyces variotii.
Fungi colonize habitats by means of spores. These cells are stress-resistant compared with growing fungal cells. Fungal conidia, asexual spores, formed by cosmopolitan fungal genera like Penicillium, Aspergillus and Peacilomyces are dispersed by air. They are present in places where food products are stored and as a result, they cause food spoilage. Here, we determined the heterogeneity of heat resistance of conidia between and within strains of Paecilomyces variotii, a spoiler of foods such as margarine, fruit juices, canned fruits and non-carbonized sodas. Out of 108 strains, 31 isolates showed a conidial survival >10% after a 10-min-heat treatment at 59°C. Three strains with different conidial heat resistance were selected for further phenotyping. Conidia of DTO 212-C5 and DTO 032-I3 showed 0.3% and 2.6% survival in the screening respectively, while survival of DTO 217-A2 conidia was >10%. The decimal reduction times of these strains at 60°C (D60 value) were 3.7 ± 0.08, 5.5 ± 0.35 and 22.9 ± 2.00 min respectively. Further in-depth analysis revealed that the three strains showed differences in morphology, spore size distributions, compatible solute compositions and growth under salt stress. Conidia of DTO 217-A2 are the most heat-resistant reported so far. The ecological consequences of this heterogeneity of resistance, including food spoilage, are discussed.


Association of health anxiety, fatalism and medication adherence among geriatric clients: An exploratory study.
This study explored the relationship between health anxiety, fatalistic beliefs, and medication adherence among geriatric clients. Also, it determines the extent to which health anxiety and fatalism can predict the variance in medication adherence among the same population of geriatric clients. A cross-sectional analytical survey on 200 eligible participants using the Arabic Version of the Short Health Anxiety Inventory, Fatalism Scale, and Morisky Medication Adherence Scale-8 items. The study found a statistically significant negative relationship between the studied geriatric clients' fatalism and health anxiety and their medication adherence (r = -0.160, - 0.187, and P = 0.024, 0.008), respectively. This study highlights the importance of considering psychological factors such as health anxiety and fatalistic beliefs in addressing medication adherence among geriatric clients. By addressing these factors, healthcare providers can develop more effective strategies to improve medication adherence and ultimately improve the health outcomes of geriatric clients.


Identification of breast tumor mutations in BRCA1 that abolish its function in homologous DNA recombination.
Effects of breast cancer-associated gene 1 (BRCA1) missense mutations on the function of BRCA1 protein in DNA recombination have been little studied. In this report, we adapted a homology-directed recombination (HDR) assay to analyze the effects of BRCA1 mutations on this function. Using a HeLa-derived cell line with a genomically integrated recombination substrate, we expressed an endonuclease creating a double-stranded break in the substrate that the HDR assay scores by generation of green fluorescent protein-positive cells. By combining RNA interference (RNAi) that targets cellular BRCA1 mRNA with expression of RNAi-resistant BRCA1 mutants, we could effectively substitute selected point mutants to test these in the cellular recombination assay. We found that approximately 300 residues at both termini of the BRCA1 protein were essential for HDR. Whereas some mutations analyzed were neutral, mutations that altered any zinc-coordinating residue or generated M18T and T37R alterations were defective for recombination. This study established a robust assay system to analyze the function of BRCA1 in regulating homologous recombination, which is critical for its tumor suppressor function.


Self-organization of spiking neurons using action potential timing.
We propose a mechanism for unsupervised learning in networks of spiking neurons which is based on the timing of single firing events. Our results show that a topology preserving behavior quite similar to that of Kohonen's self-organizing map can be achieved using temporal coding. In contrast to previous approaches, which use rate coding, the winner among competing neurons can be determined fast and locally. Our model is a further step toward a more realistic description of unsupervised learning in biological neural systems. Furthermore, it may provide a basis for fast implementations in pulsed VLSI (very large scale integration).


Does [TcF(CO)5] exist? The crystal and molecular structure of [Tc(CO)3(OH)0.49F0.51]4·[Tc(CO)5(BF4)].
Technetium pentacarbonyl fluoride [TcF(CO)5] was prepared for the first time by reaction of [TcI(CO)5] with solid AgF in a dichloromethane solution at -23 °C. Low temperature crystallization of the resulting compound in a glass vial yielded an unusual complex [Tc(CO)3(OH)0.49F0.51]4·[Tc(CO)5(BF4)] characterized by single crystal XRD.


Vertebrate hosts and vectors of Trypanosoma rangeli in the Amazon Basin of Brazil.
A total of 46 Trypanosoma rangeli stocks were isolated from naturally infected mammals and triatomine vectors. Twenty-two stocks were from the common opossum (Didelphis marsupialis), one from the brown "4-eyed" opossum (Metachirus nudicaudatus), one from the anteater (Tamandua tetradactyla), one from the coati (Nasua nasua), seven from Rhodnius pictipes and 14 from Rhodnius robustus. Two stocks were also isolated from recently fed sandflies (Lutzomyia sp., Shannoni group). The stocks were identified as T. rangeli on the basis of natural or experimental salivary gland infections in Rhodnius, inoculative (anterior station) transmission to mice, morphological parameters in parasitemic mice and comparisons of isozyme profiles with a known stock of T. rangeli isolated from man. Three other trypanosome stocks from D. marsupialis, T. tetradactyla and the three-toed sloth (Bradypus tridactylus) were morphologically similar to T. rangeli in culture but had quite different isozyme profiles and were not identified. It is concluded that T. rangeli is widely distributed in Amazonas, Pará and Rondonia States of Brazil, and probably extends into other regions where R. pictipes and R. robustus are known to occur. R. pictipes is light-attracted into houses and occasionally transmits Chagas' disease to man. It is likely that T. rangeli is also occasionally transmitted to man in the Amazon basin.


Hyperferritinemia and inflammation.
Understanding of ferritin biology has traditionally centered on its role in iron storage and homeostasis, with low ferritin levels indicative of deficiency and high levels indicative of primary or secondary hemochromatosis. However, further work has shown that iron, redox biology and inflammation are inexorably linked. During infection, increased ferritin levels represent an important host defense mechanism that deprives bacterial growth of iron and protects immune cell function. It may also be protective, limiting the production of free radicals and mediating immunomodulation. Additionally, hyperferritinemia is a key acute-phase reactants, used by clinicians as an indication for therapeutic intervention, aimed at controlling inflammation in high-risk patients. One school of thought maintains that hyperferritinemia is an 'innocent bystander' biomarker of uncontrolled inflammation that can be used to gauge effectiveness of intervention. Other schools of thought maintain that ferritin induction could be a protective negative regulatory loop. Others maintain that ferritin is a key mediator of immune dysregulation, especially in extreme hyperferritinemia, via direct immune-suppressive and pro-inflammatory effects. There is a clear need for further investigation of the role of ferritin in uncontrolled inflammatory conditions both as a biomarker and mediator of disease because its occurrence identifies patients with high mortality risk and its resolution predicts their improved survival.


Modified apparatus for collection of free-flow urine from mares.
An apparatus for collection of free-flow urine from female horses is described. This apparatus is a modification of a similar device described previously. It allows the collection of voided urine from adult female horses without confining them to metabolic stables or resorting to invasive procedures like bladder catheterisation. It is relatively easy to use after a short adaptation period.


Transcriptional, Functional, and Mechanistic Comparisons of Stem Cell-Derived Hepatocytes, HepaRG Cells, and Three-Dimensional Human Hepatocyte Spheroids as Predictive In Vitro Systems for Drug-Induced Liver Injury.
Reliable and versatile hepatic in vitro systems for the prediction of drug pharmacokinetics and toxicity are essential constituents of preclinical safety assessment pipelines for new medicines. Here, we compared three emerging cell systems-hepatocytes derived from induced pluripotent stem cells, HepaRG cells, and three-dimensional primary human hepatocyte (PHH) spheroids-at transcriptional and functional levels in a multicenter study to evaluate their potential as predictive models for drug-induced hepatotoxicity. Transcriptomic analyses revealed widespread gene expression differences between the three cell models, with 8148 of 17,462 analyzed genes (47%) being differentially expressed. Expression levels of genes involved in the metabolism of endogenous as well as xenobiotic compounds were significantly elevated in PHH spheroids, whereas genes involved in cell division and endocytosis were significantly upregulated in HepaRG cells and hepatocytes derived from induced pluripotent stem cells, respectively. Consequently, PHH spheroids were more sensitive to a panel of drugs with distinctly different toxicity mechanisms, an effect that was amplified by long-term exposure using repeated treatments. Importantly, toxicogenomic analyses revealed that transcriptomic changes in PHH spheroids were in compliance with cholestatic, carcinogenic, or steatogenic in vivo toxicity mechanisms at clinically relevant drug concentrations. Combined, the data reveal important phenotypic differences between the three cell systems and suggest that PHH spheroids can be used for functional investigations of drug-induced liver injury in vivo in humans.


Effect of low-level BCR-ABL1 kinase domain mutations identified by next-generation sequencing in patients with chronic myeloid leukaemia: a population-based study.
Kinase domain mutations in BCR-ABL1 are associated with resistance to tyrosine kinase inhibitors in patients with chronic myeloid leukaemia. Next-generation sequencing (NGS) allows detection of low-level kinase domain mutations, but its relevance in clinical practice remains debated. We aimed to examine the clinical effects of low-level kinase domain mutations identified using NGS in patients with chronic myeloid leukaemia. In this population-based study, we included consecutive patients newly diagnosed with chronic myeloid leukaemia treated with first-line tyrosine kinase inhibitors, and patients identified at the time of resistance to first-line treatment with imatinib at six institutions (teaching hospitals and district hospitals) in southeast England. We screened patients for BCR-ABL1 kinase domain mutations using NGS, irrespective of patient response to tyrosine kinase inhibitor therapy. When we detected a mutation with NGS, we retrospectively analysed all previous samples to establish the date of first occurrence and subsequent kinetics of the mutant subclone (or subclones). The primary endpoints of this study were progression-free and event-free survival at 5 years. Between Feb 1, 2007, and Dec 31, 2014, we screened 121 patients with chronic myeloid leukaemia for BCR-ABL1 kinase domain mutation. 99 consecutive patients were newly diagnosed, with available sequential RNA stored. The remaining 22 patients were diagnosed between June 1, 1999, and June 30, 2006, and were screened at the time of resistance to first-line treatment with imatinib. Imatinib was the first-line treatment for 111 patients, nilotinib for seven patients, and dasatinib for three patients. We detected a kinase domain mutation in 25 (21%) of 121 patients. Low-level kinase domain mutations were first identified in 17 (68%) of 25 patients with mutation. For patients with a complete cytogenetic response, 13 (14%) of 93 patients screened had a mutation. Five (71%) of the seven patients with a clinically relevant mutation lost complete cytogenetic response compared with 15 (17%) of 86 patients without a clinically relevant mutation (80 patients without mutation and six patients with a tyrosine kinase inhibitor-sensitive mutation, p=0·0031). Patients harbouring a mutant clone had poorer 5-year progression-free survival (65·3% [95% CI 40·5-81·8] vs 86·9% [75·8-93·2]; p=0·0161) and poorer 5-year event-free survival (22·2% [CI 5·6-45·9] vs 62·0% [50·4-71·6]; p<0·0001) than did patients without a mutation. We identified a kinase domain mutation in four (10%) of 41 patients with samples available at 3 months after starting first-line tyrosine kinase inhibitor treatment; all four subsequently progressed to accelerated phase disease compared with only three (8%) of 37 without a mutation (p<0·0001). NGS reliably and consistently detected early appearance of kinase domain mutations that would not otherwise be detected by Sanger sequencing. For the first time, to our knowledge, we report the presence of kinase domain mutations after only 3 months of therapy, which could have substantial clinical implications. NGS will allow early clinical intervention and our findings will contribute to the establishment of new recommendations on the frequency of kinase domain mutation analysis to improve patient clinical care. None.


TRAP/SMCC/mediator-dependent transcriptional activation from DNA and chromatin templates by orphan nuclear receptor hepatocyte nuclear factor 4.
The orphan nuclear receptor hepatocyte nuclear factor 4 (HNF-4) regulates the expression of many liver-specific genes both during development and in the adult animal. Towards understanding the molecular mechanisms by which HNF-4 functions, we have established in vitro transcription systems that faithfully recapitulate HNF-4 activity. Here we have focused on the coactivator requirements for HNF-4, especially for the multicomponent TRAP/SMCC/Mediator complex that has emerged as the central regulatory module of the transcription apparatus. Using a system that has been reconstituted from purified transcription factors, as well as one consisting of unfractionated nuclear extract from which TRAP/SMCC/Mediator has been depleted by specific antibodies, we demonstrate a strong dependence of HNF-4 function on this coactivator. Importantly, we further show a TRAP/SMCC/Mediator-dependence for HNF-4 transcriptional activation from chromatin templates. The latter involves cooperation with the histone acetyltransferase-containing coactivator p300, in accord with a synergistic mode of action of the two divergent coactivators. We also show that HNF-4 and TRAP/SMCC/Mediator can interact physically. This interaction likely involves primary HNF-4 activation function 2 (AF-2)-dependent interactions with the TRAP220 subunit of TRAP/SMCC/Mediator and secondary (AF-2-independent) interactions with TRAP170/RGR1. Finally, recruitment experiments using immobilized templates strongly suggest that the functional consequences of the physical interaction probably are manifested at a postrecruitment step in the activation pathway.


Involvement of Opioid System, TRPM8, and ASIC Receptors in Antinociceptive Effect of Arrabidaea brachypoda (DC) Bureau.
Arrabidaea brachypoda (DC) Bureau is a medicinal plant found in Brazil. Known as "cipó-una", it is popularly used as a natural therapeutic agent against pain and inflammation. This study evaluated the chemical composition and antinociceptive activity of the dichloromethane fraction from the roots of A. brachypoda (DEAB) and its mechanism of action. The chemical composition was characterized by high-performance liquid chromatography, and this fraction is composed only of dimeric flavonoids. The antinociceptive effect was evaluated in formalin and hot plate tests after oral administration (10-100 mg/kg) in male Swiss mice. We also investigated the involvement of TRPV1 (transient receptor potential vanilloid 1), TRPA1 (transient receptor potential ankyrin 1), TRPM8 (transient receptor potential melastatin 8), and ASIC (acid-sensing ion channel), as well as the opioidergic, glutamatergic, and supraspinal pathways. Moreover, the nociceptive response was reduced (30 mg/kg) in the early and late phase of the formalin test. DEAB activity appears to involve the opioid system, TRPM8, and ASIC receptors, clearly showing that the DEAB alleviates acute pain in mice and suggesting the involvement of the TRPM8 and ASIC receptors and the opioid system in acute pain relief.


Multifunctional, Low Friction, Antimicrobial Approach for Biomaterial Surface Enhancement.
Poly(vinyl chloride) (PVC) biomaterials perform a host of life-saving and life-enhancing roles when employed as medical devices within the body. High frictional forces between the device surface and interfacing tissue can, however, lead to a host of complications including tissue damage, inflammation, pain, and infection. We herein describe a versatile surface modification method using multifunctional hydrogel formulations to increase lubricity and prevent common device-related complications. In a clinically relevant model of the urinary tract, simulating the mechanical and biological environments encountered in vivo, coated candidate catheter surfaces demonstrated significantly lower frictional resistance than uncoated PVC, with reductions in coefficient of friction values of more than 300-fold due to hydration of the surface-localized polymer network. Furthermore, this significant lubrication capacity was retained following hydration periods of up to 28 days in artificial urine at pH 6 and pH 9, representing the pH of physiologically normal and infected urine, respectively, and during 200 repeated cycles of applied frictional force. Importantly, the modified surfaces also displayed excellent antibacterial activity, which could be facilely tuned to achieve reductions of 99.8% in adherence of common hospital-acquired pathogens, Staphylococcus aureus and Proteus mirabilis, relative to their uncoated counterparts through incorporation of chlorhexidine in the coating matrix as a model antiseptic. The remarkable, and pH-independent, tribological performance of these lubricious, antibacterial, and highly durable surfaces offers exciting promise for use of this PVC functionalization approach in facilitating smooth and atraumatic insertion and removal of a wide range of medical implants, ultimately maintaining user health and dignity.


[Spatial frequency tuning characteristics of cat primary visual cortex at different topological locations by optical imaging].
Using optical imaging based on intrinsic signals, we studied spatial frequency tuning characteristics of cat primary visual cortex at different visual topological locations. We found that the areas representing the peripheral visual field had null or very weak responses to high spatial frequency grating stimuli, whereas the areas representing the central visual field responded to the stimuli of a wide range of spatial frequencies with greater responses to high spatial frequencies. The more centered the corresponding visual field of the cortical area, the higher the cutoff spatial frequency. The spatial frequency tuning curves also tended to have a smooth shift along the cortical surface. The results above demonstrate that spatial frequency tuning characteristics of the primary visual cortex change according to different visual topological locations of the cortical areas, in addition to the existence of spatial frequency columns. It is suggested that the distribution of spatial frequency columns may be related to visual topology.