Inhibition of calcineurin facilitates the induction of memory for sensitization in Aplysia: requirement of mitogen-activated protein kinase.
The induction of both synaptic plasticity and memory is thought to depend on the balance between opposing molecular regulatory factors, such as protein kinases and phosphatases. Here we show that inhibition of protein phosphatase 2B (calcineurin, CaN) facilitates the induction of intermediate-term memory (ITM) and long-term memory (LTM) for tail shock-induced sensitization in Aplysia without any effect on short-term memory. To identify the molecular cascade underlying the improvement of memory by inhibition of CaN, we examined the role of extracellular signal-regulated kinase 1/2/mitogen-activated protein kinase (MAPK). Molecular experiments revealed that one pulse of serotonin, which by itself does not activate MAPK, leads to significant MAPK activation in the sensory neurons of the pleural ganglia when CaN is inhibited. Extending these observations, behavioral experiments showed that the facilitated induction of ITM and LTM produced by CaN inhibition depends on MAPK activity. These results demonstrate: (i) that CaN acts as an inhibitory constraint in the formation of long-lasting phases of memory, and (ii) that facilitated induction of ITM and LTM by CaN inhibition requires MAPK activity.


Combination therapy in asthma: a review.
Asthma can be defined as a chronic inflammatory disease of the airways that is reversible either spontaneously or by treatment. Despite the exponential increase in asthma research, the prevalence of asthma is on the increase, especially in children and young adults in the western societies. Inhaled therapies are the mainstay of asthma management. This is often in the form of combined therapy using two drugs in a single device to ensure adjustable maintenance dosing. Relevant literature was reviewed using available medical journals, MEDLINE, Pubmed and Science direct via the Internet. The key words employed were: asthma, combination therapy, long acting beta agonists and corticosteroids. British Thoracic Society and The National Heart, Lung and Blood Institute websites were also used in sourcing information. Several studies have shown that combination therapy using long acting beta agonists (LABA) and inhaled corticosteroids (ICS) in a single inhaler device confers complementary and synergistic effect in the management of asthma. It further improves patient compliance and reduces the complexity of treatment and morbidity associated with the disease. Recent studies have shown the combination therapy to serve not only as maintenance but also a reliever therapy with same efficacy as the short acting beta agonists (SABA). This review was able to show the advantages of using combination therapy in asthma patients. This has been a subject of review at both national and international levels as there is no single medication that is effective against both the inflammatory and bronchoconstrictive components of this disorder. Recent studies have shown that Budesonide/formoterol in a single inhaler has been found to be effective maintenance and reliever agent in both adults and children. It has also been found to be safe and more efficacious than fixed-dosing. In addition to convenience and patient compliance, combination devices also help towards individualized approach to asthma management and reduce the complexity of treatment; this appears ideal for adoption by the primary care physician with a view for the patient to effectively achieve control of his own condition.


DNA structure control by polycationic species: polyamine, cobalt ammines, and di-metallo transition metal chelates.
Many polycationic species bind to DNA and induce structural changes. The work reported here is the first phase of a program whose long-term aim is to create a class of simple and inexpensive sequence-selective compounds that will enable enhanced DNA structure control for a wide range of applications. Three classes of molecule have been included in this work: the polyamine spermine (charge: 4(+)) and spermidine (charge: 3(+)) (which are known to induce a wide range of DNA conformational changes but whose binding modes are still not well understood); cobalt (III) amine transition metal complexes as potential polyamine mimics and [Fe(H(2)O)(6)](3+); and the first member of a new class of di-metallo tris-chelated cylinders of helical structure (charge 4(+)). Temperature-dependent absorption, circular dichroism, linear dichroism, gel electrophoresis, and molecular modeling data are presented. The cobalt amines prove to be effective polyamine mimics, although their binding appears to be restricted to backbone and major groove. All the ligands stabilize the DNA, but the 4(+) di-iron tris-chelate does so comparatively weakly and seems to have a preference for single-stranded DNA. All the molecules studied bend the DNA, with the di-iron tris-chelate having a particularly dramatic effect even at very low drug load.


Controlled release of 5-fluorouracil and progesterone from magnetic nanoaggregates.
The potential use of magnetic nanoparticles in biomedical applications has witnessed an exponential growth in recent years. In this study, we used nanoaggregates of magnetic nanoparticles as carriers for controlled drug delivery. The nanoaggregates are formed due to the presence of the block copolymer of polyethylene oxide-polypropylene oxide (Pluronic F-68) and beta-cyclodextrin that surround the magnetic core of the nanoparticles. The administration of the drug carriers occurs by inhalation, and the drug is delivered systemically via the pulmonary route. We tested the delivery of 5-fluorouracil and progesterone, which are used as models of hydrophilic and hydrophobic drugs, respectively. The estimated nanoaggregates' diameters are between 293 nm ± 14.65 nm and 90.2 nm ± 4.51 nm, respectively. In-situ and post-synthesis techniques are two approaches for drug loading. The polymer composition of nanoaggregates and initial drug concentration showed a significant effect on both the drug entrapment efficiency and release kinetics. Average drug entrapment efficiencies ranged between 16.11% and 83.25%. In-situ loaded samples showed significantly slower release rates. The drug release mechanism is investigated by mathematical curve fitting to different drug release kinetics models. In most cases, the Peppas model has shown good correlations (coefficients of correlation, R(2), between 0.85 and 0.99) with the examined release profiles. The estimated release indices are below 0.5, which indicates the Fickian diffusion mechanism. For samples with an initial burst effect, the modified Peppas model can provide a better understanding of the drug release mechanism, both in the samples loaded with progesterone, or those high polymer concentrations. Our work showed prolonged delivery of drugs (5-fluorouracil and progesterone) by diffusion from nanoaggregates, with the potential to reduce dose-related adverse effects.


Correlations between short- and long-time relaxation in colloidal supercooled liquids and glasses.
Spatiotemporal dynamics of short- and long-time structural relaxation are measured experimentally as a function of packing fraction, ϕ, in quasi-two-dimensional colloidal supercooled liquids and glasses. The relaxation times associated with long-time dynamic heterogeneity and short-time intracage motion are found to be strongly correlated and to grow by orders of magnitude with increasing ϕ toward dynamic arrest. We find that clusters of fast particles on the two timescales often overlap, and, interestingly, the distribution of minimum-spatial-separation between closest nonoverlapping clusters across the two timescales is revealed to be exponential with a decay length that increases with ϕ. In total, the experimental observations suggest short-time relaxation events are very often precursors to heterogeneous relaxation at longer timescales in glassy materials.


Two monoclonal antibodies against La Crosse virus show host-dependent neutralizing activity.
Mammalian and arthropod cell cultures were used to assess the neutralizing activity of six monoclonal antibodies specific for the G1 glycoprotein of La Crosse virus. Four antibodies, two neutralizing and two non-neutralizing, showed no host-dependent differences, giving similar results when post-treatment infectivity was determined using either Aedes albopictus cells or BHK-21 cells. For two other antibodies, however, dissimilar activities were observed between the vertebrate and invertebrate cell lines. One of these antibodies was positive when BHK-21 cells were employed as the post-treatment host and negative when mosquito cells were used; the other antibody was the converse. The epitope for this last antibody was present on all California serogroup viruses examined, which suggests that it may have a special significance in the natural life-cycle of the virus.


Metabolic-digestive clinical disorders of lactating dairy cows were associated with alterations of rumination, physical activity, and lying behavior monitored by an ear-attached sensor.
The objective of this observational cohort study was to characterize the pattern of rumination time (RT), physical activity (PA), and lying time (LT) monitored by an automated health monitoring system, based on an ear-attached sensor, immediately before, during, and after clinical diagnosis (CD) of metabolic-digestive disorders. Sensor data were collected from 820 lactating Holstein cows monitored daily from calving up to 21 DIM for detection of health disorders (HD). Cows were grouped retrospectively in the no-clinical health disorder group (NCHD; n = 616) if no HD were diagnosed, or the metabolic-digestive group (METB-DIG; n = 58) if diagnosed with clinical ketosis or indigestion only. Cows with another clinical health disorder within -7 to +7 d of CD of displaced abomasum, clinical ketosis, or indigestion were included in the metabolic-digestive plus one group (METB-DIG+1; n = 25). Daily RT, PA, and LT, and absolute and relative changes within -7 to +7 d of CD were analyzed with linear mixed models with or without repeated measures. Rumination time and PA were smaller, and LT was greater for the METB-DIG and METB-DIG+1 group than for cows in the NCHD group for most days from -7 to +7 d of CD of HD. In general, daily RT, PA, and LT differences were larger between the METB-DIG+1 and NCHD groups than between the METB-DIG and NCHD groups. In most cases, RT and PA decreased to a nadir and LT increased to a peak immediately before or after CD of HD, with a return to levels similar to the NCHD group within 7 d of CD. Absolute values and relative changes from 5 d before CD to the day of the nadir for RT and PA or peak for LT were different for cows in the METB-DIG and METB-DIG+1 group than for the NCHD group. For PA, the METB-DIG+1 group had greater changes than the METB-DIG group. For cows affected by metabolic-digestive disorders, RT, PA, and LT on the day of CD and resolution of clinical signs were different than for cows in the NCHD group, but an increase in RT and PA or a decrease in LT was observed from the day of CD to the day of resolution of clinical signs. We conclude that dairy cows diagnosed with metabolic-digestive disorders including displaced abomasum, clinical ketosis, and indigestion presented substantial alterations in the pattern of RT, PA, and LT captured by an ear-attached sensor. Thus, automated health monitoring systems based on ear-attached sensors might be used as an aid for identifying cows with metabolic-digestive disorders. Moreover, RT, PA, and LT changes after CD might be positive indicators of recovery from metabolic-digestive disorders.


Validity and reliability of a new instrument to measure cancer-related fatigue in adolescents.
Adolescents undergoing treatment for cancer rate fatigue as their most prevalent and intense cancer- and treatment-related effect. Parents and staff rate it similarly. Despite its reported prevalence, intensity, and distressing effects, cancer-related fatigue in adolescents is not routinely assessed during or after cancer treatment. We contend that the insufficient clinical attention is primarily due to the lack of a reliable and valid self-report instrument with which adolescent cancer-related fatigue can be measured. Our aim was to determine the reliability and construct validity of a new instrument and its ability to measure change in fatigue over time. Initial testing involved 64 adolescents undergoing curative treatment of cancer who completed the Fatigue Scale-Adolescent (FS-A) at two to four key points in treatment in one of four studies. Internal consistency estimates ranged from 0.67 to 0.95. Validity estimates involving the FS-A with the parent version ranged from 0.13 to 0.76; estimates involving the staff version and the Reynolds Depression Scale were 0.27 and 0.87, respectively. Additional validity findings included significant fatigue differences between anemic and nonanemic patients (P=0.042) and the emergence of four factors in an exploratory factor analysis. Findings further indicate that the FS-A can be used to measure change over time (t=2.55, P<0.01). In summary, the FS-A has moderate to strong reliability and impressive validity coefficients for a new research instrument.


Influence of damaging and wilting red clover on lipid metabolism during ensiling and in vitro rumen incubation.
This paper describes the relationship between protein-bound phenols in red clover, induced by different degrees of damaging before wilting and varying wilting duration, and in silo lipid metabolism. The ultimate effect of these changes on rumen biohydrogenation is the second focus of this paper. For this experiment, red clover, damaged to different degrees (not damaged (ND), crushing or frozen/thawing (FT)) before wilting (4 or 24 h) was ensiled. Different degrees of damaging and wilting duration lead to differences in polyphenol oxidase (PPO) activity, measured as increase in protein-bound phenols. Treatment effects on fatty acid (FA) content and composition, lipid fractions (free FAs, membrane lipids (ML) and neutral fraction) and lipolysis were further studied in the silage. In FT, red clover lipolysis was markedly lower in the first days after ensiling, but this largely disappeared after 60 days of ensiling, regardless of wilting duration. This suggests an inhibition of plant lipases in FT silages. After 60 days of ensiling no differences in lipid fractions could be found between any of the treatments and differences in lipolysis were caused by reduced FA proportions in ML of wilted FT red clover. Fresh, wilted (24 h) after damaging (ND or FT) and ensiled (4 or 60 days; wilted 24 h; ND or FT) red clover were also incubated in rumen fluid to study the biohydrogenation of C18:3n-3 and C18:2n-6 in vitro. Silages (both 60 days and to a lower degree 4 days) showed a lower biohydrogenation compared with fresh and wilted forages, regardless of damaging. This suggests that lipids in ensiled red clover were more protected, but this protection was not enhanced by a higher amount of protein-bound phenols in wilted FT compared with ND red clover. The reduction of rumen microbial biohydrogenation with duration of red clover ensiling seems in contrast to what is expected, namely a higher biohydrogenation when a higher amount of FFA is present. This merits further investigation in relation to strategies to activate PPO toward the embedding of lipids in phenol-protein complexes.


Absence of circulating adrenal autoantibodies in adult-onset X-linked adrenoleukodystrophy.
X-linked adrenoleukodystrophy (ALD) is a frequent cause of adrenal insufficiency in young-adult patients with Addison's disease. As the contribution of an autoimmune process in the destruction of steroid cells in ALD is unclear, the aim of the present study was to evaluate the occurrence of adrenal-, thyroid- and islet-specific and non organ-specific autoantibodies in adult ALD patients. In all 5 patients, Addison's disease was the first manifestation of ALD. None of the ALD patients were positive for adrenal cortex autoantibodies in an indirect immunofluorescence assay, or for 21-hydroxylase autoantibodies in a radiobinding assay with recombinant human antigen. Similarly, we found neither non-organ specific autoantibodies (such as anti-nuclear, anti-ribosomal, anti-mitochondria, anti-smooth-muscle, anti-liver/kidney microsomal or anti-reticulin autoantibodies), nor islet-cell antibodies or glutamic acid decarboxylase (GAD65 or GAD67) autoantibodies, nor thyroglobulin autoantibodies in the sera of the 5 ALD patients. Two out of five patients were positive for thyroid microsomal autoantibodies. One of the two latter thyroid antibody-positive patients had clinical symptoms of hypothyroidism, and the other presented high levels of circulating TSH but no clinical signs or symptoms of hypothyroidism. Our study demonstrates that adult ALD is not immediately associated with the presence of adrenal autoantibodies and suggests that adrenal insufficiency is not mediated by an autoimmune process in adult ALD patients.


Generic method for attaching biomolecules via avidin-biotin complexes immobilized on films of regenerated and nanofibrillar cellulose.
We investigated the adsorption and chemical conjugation of avidin and its deglycosylated form, neutravidin, on films of regenerated and nanofibrillar cellulose. The dynamics and extent of biomolecular attachment were monitored in situ by quartz crystal microbalance microgravimetry and ex situ via surface analyses with atomic force microscopy and X-ray photoelectron spectroscopy. The installation of carboxyl groups on cellulose after modification with carboxymethylated cellulose (CMC) or TEMPO-oxidation significantly increases physisorption of avidins, which can be then covalently conjugated by using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride/N-hydroxysuccinimide (EDS/NHS) coupling chemistries. The developed cellulose-avidin biointerfaces are able to scavenge biotinylated molecules from solution as demonstrated by successful surface complexation of biotinylated bovine serum albumin (Biotin-BSA) and antihuman immunoglobulin G (Biotin-anti-hIgG). Finally, we show that cellulose substrates carrying immobilized anti-hIgG are effective in detecting human immunoglobulin G (hIgG) from fluid matrices.


The role of excitotoxic injury in post-traumatic syringomyelia.
Fifty percent of patients with neurological deterioration from post-traumatic syringomyelia do not respond to treatment. Treatment failure is due in part to an incomplete understanding of the underlying aetiology. An animal model that mimics the human disease is required to investigate underlying pathophysiology and treatment options. A previous study was designed to mimic trauma-induced effects on the spinal cord that result in syringomyelia, combining an excitotoxic insult with kaolin-induced arachnoiditis. In this excitotoxic model, syringes were produced in 82% of animals. The aims of the current study were to improve the model to produce syringes in all animals treated, to examine the relative influences of excitotoxic injury and neuronal loss on syrinx formation, and to use magnetic resonance imaging (MRI) to examine syringes non-invasively. A temporal and dose profile of intraparenchymal quisqualic acid (QA) and subarachnoid kaolin was performed in Sprague Dawley rats. MRI was used to study four syrinx and six control animals. In one subgroup of animals surviving for 6 weeks, 100% (eight of eight) developed syringes. Syrinx formation and enlargement occurred in a dose and time dependent manner, whilst significant neuronal loss was only dose dependent. Animal syrinx histology closely resembled human post-traumatic syringomyelia. Axial T2-weighted MR images demonstrated syrinx presence. The results suggest that the formation of an initial cyst predisposes to syrinx formation in the presence of subarachnoid adhesions.


A fast gas chromatography/mass spectrometry method for the determination of stimulants and narcotics in urine.
A fast method has been developed for the simultaneous determination of 52 stimulants and narcotics excreted unconjugated in urine by gas chromatography/mass spectrometry (GC/MS). The procedure involves the liquid/liquid extraction of the analytes from urine at strong alkaline pH and the injection of the extract into a GC/MS instrument with a fast GC column (10 m x 0.18 mm i.d.); the short column allows the complete separation of the 52 analytes in a chromatographic run of 8 min. The method has been fully validated giving lower limits of detection (LLODs) satisfactory for its application to antidoping analysis as well as to forensic toxicology. The repeatability of the concentrations and the retention times are good both for intra- and for inter-day experiments (%CV of concentrations always lower than 15 and %CV of retention times lower than 0.6). In addition, the analytical bias is satisfactory (A% always >15%). The method proposed here would be particularly useful whenever there are time constraints and the analyses have to be completed in the shortest possible time.


Control of the arteriolar myogenic response by transvascular fluid filtration.
Mechanisms of the myogenic response have not been completely established. We hypothesized that transvascular fluid filtration from plasma across smooth muscle cells (SMC) and into the surrounding interstitium helps regulate arteriolar myogenic tone. Arteriolar diameters in the rat mesentery were monitored before and following vascular occlusion with a glass micropipette. Arteriolar occlusion not only gave an increase in hydrostatic pressure that initiated myogenic constriction upstream of the pipette, but also allowed measurement of fluid filtration rate by monitoring the movement of vascular red blood cells. A statistically significant correlation (P < 0.001) existed between basal myogenic tone and fluid filtration. Additionally, the myogenic response was attenuated by 47% +/- 7% (N = 10) when an osmotic solution of albumin or albumin plus Ficoll was infused into the bloodstream to decrease fluid filtration by 53% +/- 3%. Moreover, the same inhibition of myogenic tone was found in isolated, cannulated rat soleus muscle arterioles when filtration was osmotically attenuated by intravascular dextran. Taken together, these results are consistent with the hypothesis that shear stress on arteriolar smooth muscle, induced by transvascular fluid filtration, is a contributing factor that helps control myogenic tone.


Comparative evaluation of meniscal pathology: MRI vs arthroscopy.
The meniscal pathology of the knee is one of the clinical realities the orthopedic surgeon must daily confront with. The diagnosis is generally both clinical and instrumental; among the different diagnostic imaging techniques, Magnetic Resonance Imaging (MRI) appears to be the most accurate method regarding sensitivity and specificity for the study of meniscal fibrocartilages and articular cartilage. In an attempt to clarify the roles of MRI and diagnostic knee arthroscopy, we performed a retrospective comparative study of the two methods to assess their sensitivity and specificity in the diagnosis of meniscal pathology. We evaluated 105 consecutive patients with a clinical diagnosis of intra-articular knee pathology who were subjected to MRI examination and subsequently to surgical arthroscopy, recording on a graphic card the surgical and radiographic findings expressed by a blinded expert radiologist. Comparison of MRI and arthroscopy data showed, for the internal meniscus, values of 98.5% sensitivity, 94.7% specificity and 93.8% "K" index for MRI compared to arthroscopy, and of 90%, 98.6% and 90.5% for the external meniscus. These results allow us to state that the diagnostic capacity of MRI appears to be very high and therefore crucial in the planning of the correct surgical treatment of individual patients, thanks to its ability to highlight even small changes affecting intra-articular structures.


[Association of pulmonar tuberculosis with HLA system antigens in Northeastern Mexico].
Genetic susceptibility to pulmonary tuberculosis (PTb) has been associated with the HLA (Antigens of the Human Leukocytes) system of the MHC (Major Histocompatibility Complex), mainly with HLA-DR and-DQ antigens. Based on this assumption we carried out a case control study to determine the association of PTb with the HLA-DR and-DQ antigens among a sample of patients attending a medical unit belonging to the Mexican Social Security System (IMSS). HLA system phenotypes from cases (n=50) and controls (n=417), were defined serologically using a complement dependent microlymphocytotoxic assay. B lymphocytes were obtained using immunobeads. The allele and haplotype frequencies were determined using the Arlequin version 3.01 computer software. Relative risk (RR) was calculated with the Epimax Table Calculator. The alelles HLA-DR11(5), -DR16(2) and -DQ7(3) and haplotypes /DR11(5)-DQ7(3), /DR14(6)-DQS(1) and /DR16(2)-DQ7(3) had a higher frequency in cases than in controls (RR>1, p<0.05). The HLA-DR17(3) and DQ8(3) alelles and /DR17(3)-DQ2 and /DR4-DQ8(3) haplotypes had a higher frequency among controls than among cases (RR<1, p<.05). These results indicate an association between PTb with the HLA-DR and -DQ antigens in a Mexican sample. Our results are similar to those found in the international literature.


The Lycopodium alkaloids.
Lycopodium alkaloids are quinolizine, or pyridine and alpha-pyridone type alkaloids. Some Lycopodium alkaloids are potent inhibitors of acetylcholinesterase (AChE). Huperzine A (HupA) is reported to increase efficiency for learning and memory in animals, and it shows promise in the treatment of Alzheimer's disease (AD). 201 Lycopodium alkaloids from 54 species of Lycopodium (sensu lato) have been reported so far. This review is intended to to cover the chemical, pharmacological and clinical research on Lycopodium alkaloids reported in the literature from the spring of 1993 to August 2004. Structures of 81 new Lycopodium alkaloids are presented, classified and analyzed. The structural characters and biogenetic relationships of the four major Lycopodium alkaloid groups (lycopodine, lycodine, fawcettimine and miscellaneous) are discussed. Bioactivities of Lycopodium alkaloids, especially HupA, are summarized. In particular, the effect of HupA and other cholinesterase inhibitors (anti-AD drugs) on acetylcholine esterase (AChE) activity in the rat cortex and butylcholine esterase activity are compared. Structure-activity relationships and structure modifications of HupA and its analogs are described. Information on clinical trials with HupA and its derivative ZT-1 is presented. The state of HupA availability and recent advances in in vitro propagation of HupA producing plants are outlined. Finally, hypotheses about Lycopodium alkaloid biosynthetic pathways are discussed.


[The level of blood insulin and the glycolytic activity of the blood in patients with ischemic heart disease].
Disorders of the glucose tolerance test (diabetic and borderline glycemic curves) were revealed in 49% of patients with ischemic heart disease. These patients showed hyperinsulinemia. The glycolytic activity of the blood, especially in patients with circulatory insufficiency, was increased evidencing lagging behind of the aerobic phase of carbohydrate oxidation from the glycolytic splitting. Disorders of the carbohydrate metabolism in these patients is caused by relative insulin insufficiency and tissue hypoxia.


Changes in midlife death rates across racial and ethnic groups in the United States: systematic analysis of vital statistics.
To systematically compare midlife mortality patterns in the United States across racial and ethnic groups during 1999-2016, documenting causes of death and their relative contribution to excess deaths. Trend analysis of US vital statistics among racial and ethnic groups. United States, 1999-2016. US adults aged 25-64 years (midlife). Absolute changes in mortality measured as average year-to-year change during 1999-2016 and 2012-16; excess deaths attributable to increasing mortality; and relative changes in mortality measured as relative difference between mortality in 1999 versus 2016 and the nadir year versus 2016, and the slope of modeled mortality trends for 1999-2016 and for intervals between joinpoints. During 1999-2016, all cause mortality in midlife increased not only among non-Hispanic (NH) whites but also among NH American Indians and Alaskan Natives. Although all cause mortality initially decreased among NH blacks, Hispanics, and NH Asians and Pacific Islanders, this trend ended in 2009-11. Drug overdoses were the leading cause of increased mortality in midlife in each population, but mortality also increased for alcohol related conditions, suicides, and organ diseases involving multiple body systems. Although midlife mortality among NH whites increased across a multitude of conditions, a similar trend affected non-white populations. Absolute (year-to-year) increases in midlife mortality among non-white populationsoften matched or exceeded those of NH whites, especially in 2012-16, when the rate of increase intensified for many causes of death. During 1999-2016, NH American Indians and Alaskan Natives experienced large increases in midlife mortality from 12 causes, not only drug overdoses (411.4%) but also hypertensive diseases (269.3%), liver cancer (115.1%), viral hepatitis (112.1%), and diseases of the nervous system (99.8%). NH blacks experienced increased midlife mortality from 17 causes, including drug overdoses (149.6%), homicides (21.4%), hypertensive diseases (15.5%), obesity (120.7%), and liver cancer (49.5%). NH blacks also experienced retrogression: after a period of stable or declining midlife mortality early in 1999-2016, death rates increased for alcohol related liver disease, chronic lower respiratory tract disease, suicides, diabetes, and pancreatic cancer. Among Hispanics, midlife mortality increased across 12 causes, including drug overdoses (80.0%), hypertensive diseases (40.6%), liver cancer (41.8%), suicides (21.9%), obesity (106.6%), and metabolic disorders (60.0%). Retrogression also occurred in this population; after a period of declining mortality, death rates increased for alcohol related liver disease, mental and behavioral disorders involving psychoactive substances, and homicides. NH Asians and Pacific Islanders were least affected by this trend but also experienced increases in midlife mortality from drug overdoses (300.6%), alcohol related liver disease (62.9%), hypertensive diseases (28.3%), and brain cancer (56.6%). The suicide rate in this group increased by 29.7% after 2001. The relative increase in US midlife mortality differed by sex and geography. For example, the relative increase in fatal drug overdoses was greater among women than among men. Although the relative increase in midlife mortality was generally greater in non-metropolitan (ie, rural) areas, the relative increase in drug overdoses among NH whites and Hispanics was greatest in suburban fringe areas of large cities, and among NH blacks was greatest in small cities. Mortality in midlife in the US has increased across racial-ethnic populations for a variety of conditions, especially in recent years, offsetting years of progress in lowering mortality rates. This reversal carries added consequences for racial groups with high baseline mortality rates, such as for NH blacks and NH American Indians and Alaskan Natives. That death rates are increasing throughout the US population for dozens of conditions signals a systemic cause and warrants prompt action by policy makers to tackle the factors responsible for declining health in the US.


Do Patients Taking Warfarin Experience Delays to Theatre, Longer Hospital Stay, and Poorer Survival After Hip Fracture?
Patients sustaining a fractured neck of the femur are typically of advanced age with multiple comorbidities. As a consequence, the proportion of these patients receiving warfarin therapy is approximately 10%. There are currently few studies investigating outcomes in this subset of patients. The purpose of this study was to assess the association between warfarin therapy and time to surgery, length of hospital stay, and survival in patients sustaining a fractured neck of the femur. Data for 2036 patients admitted to our center between July 2009 and July 2014 with a fractured neck of the femur were extracted from the National Hip Fracture Database. Fifty-seven patients received no surgical treatment and were excluded from analysis. Multivariable ordinary least squares regression was performed to test the association between warfarin treatment on time to surgery and length of stay, and Cox proportional hazards to test followup survival. Variables included in the regression model were age, sex, American Society of Anesthesiologists (ASA) score, admission Abbreviated Mental Test Score (AMTS), fracture type, operation type, and premorbid Work Ability Index (WAI). One hundred fifty-two of 1979 surgically treated patients (8%) were receiving warfarin therapy at the time of admission. After controlling for age, sex, ASA score, AMTS, fracture type, operation type, and WAI, we found that patients taking warfarin were less likely to go to surgery by 36 hours (odds ratio [OR], 0.20; 95% CI, 0.14-0.30), and less likely to go to surgery by 48 hours (OR, 0.17; 95% CI, 0.11-0.24). Patients taking warfarin had a longer length of stay (median, 15 days; interquartile range [IQR], 12-22 days) compared with patients not taking warfarin (median, 13 days; IQR, 9-20 days; p < 0.001). Survival analysis to June 2015 showed a higher mortality for patients taking warfarin (12-month survival, 66% vs 76%; hazard ratio, 1.57; 95% CI, 1.21-2.04; p < 0.001). After controlling for multiple prognostic factors such as age, ASA score, AMTS, and WAI, warfarin therapy at the time of injury is associated with increased time to surgery, length of stay, and decreased survival. This study highlights the need to view warfarin therapy as a 'red flag' in patients presenting with a fractured neck of the femur. Preoperatively, prompt warfarin reversal together with adequate investigation and optimization of the patient should ensure timely, safe surgery. Early involvement of the anesthesia team should ensure an appropriate level of postoperative care for these patients. Level III, therapeutic study.


Rpd3L Contributes to the DNA Damage Sensitivity of Saccharomyces cerevisiae Checkpoint Mutants.
DNA replication forks that are stalled by DNA damage activate an S-phase checkpoint that prevents irreversible fork arrest and cell death. The increased cell death caused by DNA damage in budding yeast cells lacking the Rad53 checkpoint protein kinase is partially suppressed by deletion of the EXO1 gene. Using a whole-genome sequencing approach, we identified two additional genes, RXT2 and RPH1, whose mutation can also partially suppress this DNA damage sensitivity. We provide evidence that RXT2 and RPH1 act in a common pathway, which is distinct from the EXO1 pathway. Analysis of additional mutants indicates that suppression works through the loss of the Rpd3L histone deacetylase complex. Our results suggest that the loss or absence of histone acetylation, perhaps at stalled forks, may contribute to cell death in the absence of a functional checkpoint.


Neuropeptide S promotes wakefulness through the inhibition of sleep-promoting ventrolateral preoptic nucleus neurons.
The regulation of sleep-wake cycles is crucial for the brain's health and cognitive skills. Among the various substances known to control behavioral states, intraventricular injection of neuropeptide S (NPS) has already been shown to promote wakefulness. However, the NPS signaling pathway remains elusive. In this study, we characterized the effects of NPS in the ventrolateral preoptic nucleus (VLPO) of the hypothalamus, one of the major brain structures regulating non-rapid eye movement (NREM) sleep. We combined polysomnographic recordings, vascular reactivity, and patch-clamp recordings in mice VLPO to determine the NPS mode of action. We demonstrated that a local infusion of NPS bilaterally into the anterior hypothalamus (which includes the VLPO) significantly increases awakening and specifically decreases NREM sleep. Furthermore, we established that NPS application on acute brain slices induces strong and reversible tetrodotoxin (TTX)-sensitive constriction of blood vessels in the VLPO. This effect strongly suggests that the local neuronal network is downregulated in the presence of NPS. At the cellular level, we revealed by electrophysiological recordings and in situ hybridization that NPSR mRNAs are only expressed by non-Gal local GABAergic neurons, which are depolarized by the application of NPS. Simultaneously, we showed that NPS hyperpolarizes sleep-promoting neurons, which is associated with an increased frequency in their spontaneous IPSC inputs. Altogether, our data reveal that NPS controls local neuronal activity in the VLPO. Following the depolarization of local GABAergic neurons, NPS indirectly provokes feed-forward inhibition onto sleep-promoting neurons, which translates into a decrease in NREM sleep to favor arousal.


Associations between mass media exposure and birth preparedness among women in southwestern Uganda: a community-based survey.
Exposure to mass media provides increased awareness and knowledge, as well as changes in attitudes, social norms and behaviors that may lead to positive public health outcomes. Birth preparedness (i.e. the preparations for childbirth made by pregnant women, their families, and communities) increases the use of skilled birth attendants (SBAs) and hence reduces maternal morbidity and mortality. The aim of this study was to explore the association between media exposure and birth preparedness in rural Uganda. A total of 765 recently delivered women from 120 villages in the Mbarara District of southwest Uganda were selected for a community-based survey using two-stage cluster sampling. Univariate and multivariate logistic regression was performed with generalized linear mixed models using SPSS 21. We found that 88.6% of the women surveyed listened to the radio and 33.9% read newspapers. Birth preparedness actions included were money saved (87.8%), identified SBA (64.3%), identified transport (60.1%), and purchased childbirth materials (20.7%). Women who had taken three or more actions were coded as well birth prepared (53.9%). Women who read newspapers were more likely to be birth prepared (adjusted OR 2.2, 95% CI 1.5-3.2). High media exposure, i.e. regular exposure to radio, newspaper, or television, showed no significant association with birth preparedness (adjusted OR 1.3, 95% CI 0.9-2.0). Our results indicate that increased reading of newspapers can enhance birth preparedness and skilled birth attendance. Apart from general literacy skills, this requires newspapers to be accessible in terms of language, dissemination, and cost.


Effectiveness of intraoperative bimanual palpation in metastatic tumors of lung.
In this study, we aimed to compare effectiveness of thoracic computed tomography versus intraoperative bimanual palpation in the detection of number of nodules in patients undergoing thoracotomy. Between January 2011 and January 2019, a total of 157 patients (63 males, 94 females; mean age: 46.6±11.2 years; range, 13 to 77 years) who underwent pulmonary metastasectomy in our institution were retrospectively analyzed. Metastatic nodules evaluated using thoracic computed tomography were compared with nodules detected by intraoperative palpation. A total of 226 muscle-sparing thoracotomy was performed in 157 patients. The time between the preoperative thoracic computed tomography and operation ranged from 3 to 24 days. Metastasectomy with muscle-sparing thoracotomy was performed in 41 (26%) patients two times, in eight (5%) patients three times, and in four (2.5%) patients four times due to bilateral lung metastasis or re-metastasectomy. The thoracic computed tomography could detect 476 metastatic nodules, while 1,218 nodules were palpated and resected intraoperatively. Of these nodules, 920 were pathologically evaluated as metastatic. Our study results showed that the number of nodules reported as pathologically malignant after resection was 1.9 times higher than those reported by thoracic computed tomography. This finding indicates that intraoperative bimanual examination significantly increases the possibility of complete resection. This situation raises the need for more caution for the thoracoscopic metastasectomy procedure in which there is no possibility of intraoperative bimanual palpation.


In vitro activity of levofloxacin against recent Gram-negative nosocomial pathogens.
The objective of our study was to evaluate the in vitro activity of levofloxacin in comparison with other antibiotics (coamoxiclav, ceftriaxone, ceftazidime, meropenem, aztreonam, ciprofloxacin and netilmicin) against about 500 Gram-negative nosocomial microorganisms, isolated during 2003. The bactericidal activities (minimal bactericidal activity and killing curve) of levofloxacin and the other antibiotics were also evaluated. The results confirm the excellent in vitro activity of levofloxacin against multiresistant nosocomial Gram-negative pathogens, including the 73 extended spectrum beta-lactamase producer strains (90% of Escherichia coli, Enterobacter cloacae and Klebsiella pneumoniae were inhibited at 0.5 mg/l). Of particular interest is its susceptibility and bactericidal activity against Stenotrophomonas maltophilia strains. In conclusion, due to the favorable pharmacokinetics and pharmacodynamics of the molecule, associated with its safety, levofloxacin may represent a valid therapeutic option in the treatment of severe Gram-negative nosocomial infections.


[Health, public health, society--results from the past, responsibilities for the future].
Communication is the basic precondition of a global society. At the same time, however, communication is a basic cause for today's health hazards and diseases. This is an experience of our time, it is a historical experience of social change as well. If a global society wants to survive, the existing hazards to life and health have to be understood and treated on a global scale. Apart from a "new morbidity" and chronic diseases also infectious diseases again require more and more attention even in industrialised countries. The history of public health offers an overwhelming expertise of well-established concepts for every imaginable risk, to handle infectious diseases which are either not yet or no longer controllable on an individual therapeutical level. History of medicine also shows, that the effectiveness of hygienic concepts is based on their scientific validity and the resulting predictability of their interventions. This implies, that modern hygiene has to incorporate actively the concept of molecular medicine. Every single discipline involved, i.e. hygiene/sanitation, toxicology, microbiology, bacteriology, immunology, and infectiology, has to join in interdisciplinary scientific and interventional efforts.


Identification of bacterial protease domains that cleave human IgM.
Immunoglobulin-degrading proteases are secreted by pathogenic bacteria to weaken the host immune response, contributing to immune evasion mechanisms during an infection. Proteases specific to IgG and IgA immunoglobulin classes have previously been identified and characterized, and only a single report exists on a porcine specific IgM-degrading enzyme. It is unclear whether human pathogens also produce enzymes that can break down human IgM. Here, we have identified four novel IgM-degrading proteases from different genera of human-infecting bacterial pathogens. All four protease domains cleave human IgM at a conserved and unique site in the constant region of IgM. These human IgM proteases may be a useful biochemical tool for the study of early immune responses and have therapeutic potential in IgM-mediated disease.


Characterization of an outer membrane mannanase from Bacteroides ovatus.
Bacteroides ovatus utilizes guar gum, a high-molecular-weight branched galactomannanan, as a sole source of carbohydrate. No extracellular activity was detectable. Approximately 30% of the total cell-associated mannanase activity partitioned with cell membranes. When inner and outer membranes of B. ovatus were separated on sucrose gradients, the mannanase activity was associated mainly with fractions containing outer membranes. Enzyme activity was solubilized by 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or by Triton X-100 at a detergent-to-protein ratio of 1:1. The enzyme was stable for only 4 h at 37 degrees C and for 50 to 60 h at 4 degrees C. Analysis of the products of the CHAPS-solubilized mannanase on Bio-Gel A-5M and Bio-Gel P-10 gel filtration columns indicated that the enzyme breaks guar gum into high-molecular-weight fragments. The CHAPS-solubilized mannanase was partially purified by chromatography on a FPLC Mono Q column. The partially purified mannanase preparation contained three major polypeptides (Mr 94,500, 61,000, and 43,000) and several minor ones. High mannanase activity was seen only when B. ovatus was grown on guar gum. Cross-absorbed antiserum detected two other guar gum-associated outer membrane proteins: a CHAPS-extractable 49,000-dalton polypeptide and a 120,000-dalton polypeptide that was not solubilized by CHAPS. Neither of these polypeptides was detectable in the partially purified mannanase preparation. These results indicate that there are at least two guar gum-associated outer membrane polypeptides other than the mannanase.


The effects of oxotremorine, epibatidine, atropine, mecamylamine and naloxone in the tail-flick, hot-plate, and formalin tests in the naked mole-rat (Heterocephalus glaber).
The naked mole-rat (Heterocephalus glaber) is a promising animal model for the study of pain mechanisms, therefore a thorough characterization of this species is essential. The aim of the present study was to establish the naked mole-rat as a model for studying the cholinergic receptor system in antinociception by investigating the involvement of muscarinic, nicotinic and opioid receptors in nociceptive tests in this species. The effects of systemic administration of the muscarinic receptor agonist oxotremorine and the nicotinic receptor agonist epibatidine were investigated in the tail-flick, the hot-plate, and the formalin tests. The effects of co-administration of the muscarinic receptor antagonist atropine, the nicotinic receptor antagonist mecamylamine, and the opioid receptor antagonist naloxone were also investigated. Oxotremorine and epibatidine induced a significant, dose-dependent antinociceptive effect in the tail-flick, hot-plate, and formalin tests, respectively. The effects of oxotremorine and epibatidine were blocked by atropine and mecamylamine, respectively. In all three nociceptive tests, naloxone in combination with oxotremorine or epibatidine enhanced the antinociceptive effects of the drugs. The present study demonstrated that stimulation of muscarinic and nicotinic receptors produces antinociceptive effects in the naked-mole rat. The reversal effect of atropine and mecamylamine suggests that this effect is mediated by cholinergic receptors. As naloxone increases the antinociceptive effects of cholinergic agonists, it is suggested that the cholinergic antinociception acts via a gateway facilitated by opioid receptor blockage; however, the precise interaction between these receptor systems needs further investigation.


Challenges of nurse delivery of psychological interventions for long-term conditions in primary care: a qualitative exploration of the case of chronic fatigue syndrome/myalgic encephalitis.
The evidence base for a range of psychosocial and behavioural interventions in managing and supporting patients with long-term conditions (LTCs) is now well-established. With increasing numbers of such patients being managed in primary care, and a shortage of specialists in psychology and behavioural management to deliver interventions, therapeutic interventions are increasingly being delivered by general nurses with limited training in psychological interventions. It is unknown what issues this raises for the nurses or their patients. The purpose of the study was to examine the challenges faced by non-specialist nurses when delivering psychological interventions for an LTC (chronic fatigue syndrome/myalgic encephalomyelitis [CFS/ME]) within a primary care setting. A qualitative study nested within a randomised controlled trial [ISRCTN 74156610] explored the experiences and acceptability of two different psychological interventions (pragmatic rehabilitation and supportive listening) from the perspectives of nurses, their supervisors, and patients. Semi structured in-depth interviews were conducted with three nurse therapists, three supervisors, and 46 patients. An iterative approach was used to develop conceptual categories from the dataset. Analyses identified four sets of challenges that were common to both interventions: (i) being a novice therapist, (ii) engaging patients in the therapeutic model, (iii) dealing with emotions, and (iv) the complexity of primary care. Each challenge had the potential to cause tension between therapist and patient. A number of strategies were developed by participants to manage the tensions. Tensions existed for nurses when attempting to deliver psychological interventions for patients with CFS/ME in this primary care trial. Such tensions should be addressed before implementing psychological interventions within routine clinical practice. Similar tensions may be found for other LTCs. Our findings have implications for developing therapeutic alliances and highlight the need for regular supervision.


Antigen-presenting cells and tolerance induction.
T cell tolerance induction to foreign and self-antigens has occupied research since the beginning of the understanding of the immune system. Much controversy still exists on this question even though new methods became available to investigate immunoregulatory mechanisms. Antigen-presenting cells play a pivotal role in transferring information from the periphery of the organism to lymphoid organs. There, they initiate not only the activation of naive T cells but seem to deliver important signals which result in T cell unresponsiveness with antigen-specific tolerance induction.


Verifying tumor position during stereotactic body radiation therapy delivery using (limited-arc) cone beam computed tomography imaging.
Proof of tumor position during stereotactic body radiotherapy (SBRT) delivery is desirable. We investigated if cone-beam CT (CBCT) scans reconstructed from (collimated) fluoroscopic kV images acquired during irradiation could show the dominant tumor position. Full-arc CBCT scans were reconstructed using FDK filtered back projection from 38kV fluoroscopy datasets (16 patients) continuously acquired during volumetric modulated spine SBRT. CBCT-CT match values were compared to the average spine offset values found using template matching+triangulation of the individual kV images. Multiple limited-arc CBCTs were reconstructed from fluoroscopic images acquired during lung SBRT of an anthropomorphic thorax phantom using 20-180° arc lengths and for 3 breath-hold lung SBRT patients. Differences between 3D CBCT-CT match results and average spine offsets found using template matching+triangulation were 0.1±0.1mm for all directions (range: 0.0-0.5mm). For limited-arc CBCTs of the thorax phantom, the automatic 3D CBCT-CT match results for arc lengths of 80-180° were ≤1mm. 20° CBCT reconstruction still allowed for positional verification in 2D. (Limited-arc) CBCT reconstructions of kV images acquired during irradiation can identify the dominant position of the tumor during treatment delivery.


The Impact Of Academic Medical Center Policies Restricting Direct-To-Physician Marketing On Opioid Prescribing.
Direct-to-physician opioid marketing by pharmaceutical companies is widespread and may contribute to opioid overprescribing, an important driver of the US opioid crisis. Using a difference-in-differences approach and Medicare Part D prescriber data, we examined the effects of academic medical centers' conflict-of-interest policies that restrict direct-to-physician marketing of all drugs on opioid prescribing by physicians at eighty-five centers in the period 2013-16. We examined restrictions on gifts and meals, speaking and consulting engagements, and industry representatives' access to academic medical centers, as well as rules requiring conflict-of-interest disclosures. Bans on sales representatives were associated with a 4.7 percent reduction in the total volume of opioids prescribed and disclosure requirements with a 2.5 percent reduction, while having all four marketing restriction policies was associated with an 8.8 percent reduction. Policies that restrict direct-to-physician pharmaceutical marketing may curb opioid prescribing, but additional patient-level research is needed to understand how such policies affect the delivery of evidence-based treatment for chronic pain.


Production and transport of endogenous trophic activity in a peripheral nerve following target removal.
High concentrations of trophic factor for sympathetic neurones were found in the discrete nerve which innervates the expansor secundariorum muscle of the chicken wing. Mouse nerve growth factor (NGF) was additive with nerve extract in allowing survival of sympathetic neurones. Antiserum to NGF, while inhibiting outgrowth in response to NGF, only partially blocked survival promoted by the nerve extract. In these characteristics, the nerve extract resembled the previously characterized activity of its target organ. However, after surgical removal of the muscle there was no decrease in the concentration of trophic factors in the nerve 7 days later. Likewise sectioning, crushing or removing a piece of nerve 1 cm from the muscle had little effect on trophic levels after muscle removal. Retrograde transport of the activity occurred in the nerve even in the absence of the muscle. Distal to ligatures placed on the nerve stump 24 h earlier, 60% of the trophic activity was inhibited by an antiserum to mouse NGF suggesting that a chicken form of the molecule is present and transported. In culture, cells of the nerve sheath produced trophic factors, and the rate of production increased greatly during the first 24 h of incubation. Two conclusions are made: the major source of trophic activity in the nerve is Schwann cells and at least two molecules are present, one of which is a chicken form of NGF.


The diagnostic value of miR-124a expression in peripheral blood and synovial fluid of patients with rheumatoid arthritis.
Rheumatoid arthritis (RA), a chronic autoimmune disorder, is currently a severe health threat. Previous studies have documented the altered expression of various miRNAs in RA patients. This study determined the expression of miR-124a in RA patients and estimated its diagnostic value for RA. Total 80 RA patients were enrolled as the study subjects and 36 patients with osteoarthritis were included, with another 36 healthy people as the controls. miR-124a expression levels in peripheral blood plasma, peripheral blood mononuclear cells (PBMCs), and synovial fluid were measured using RT-qPCR, followed by Pearson correlation analysis. Additionally, the association between miR-124a and major clinical indicators was assessed, such as rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), and disease activity score of 28 joints (DAS28). The diagnostic efficacy of miR-124a expression in plasma, PBMCs, and synovial fluid for RA was evaluated by the receiver operating characteristic (ROC) curve, and the difference in the area under the curve (AUC) was analyzed. miR-124a was downregulated in RA patients and the expression levels of miR-124a in plasma, PBMCs, and synovial fluid showed a certain degree of positive correlation. miR-124a was inversely linked with RF, ESR, and DAS28. For the diagnosis of RA patients, the AUC of plasma miR-124a was 0.899 and the cut-off value was 0.800, with 68.75% sensitivity and 94.44% specificity; the AUC of miR-124a in PBMCs was 0.937, and the cut-off value was 0.805, with 82.50% sensitivity and 91.67% specificity; the AUC of miR-124a in plasma combined with PBMCs was 0.961, with a higher diagnostic value than independent plasma or PBMCs; the AUC of miR-124a in synovial fluid was 0.929, and the cut-off value was 0.835, with 80.00% sensitivity and 88.89% specificity. miR-124a expression is downregulated in plasma, PBMCs, and synovial fluid of RA patients and has a high diagnostic value for RA.


Functional analysis of V3A and related areas in human visual cortex.
Using functional magnetic resonance imaging (fMRI) and cortical unfolding techniques, we analyzed the retinotopy, motion sensitivity, and functional organization of human area V3A. These data were compared with data from additional human cortical visual areas, including V1, V2, V3/VP, V4v, and MT (V5). Human V3A has a retinotopy that is similar to that reported previously in macaque: (1) it has a distinctive, continuous map of the contralateral hemifield immediately anterior to area V3, including a unique retinotopic representation of the upper visual field in superior occipital cortex; (2) in some cases the V3A foveal representation is displaced from and superior to the confluent foveal representations of V1, V2, V3, and VP; and (3) inferred receptive fields are significantly larger in human V3A, compared with those in more posterior areas such as V1. However, in other aspects human V3A appears quite different from its macaque counterpart: human V3A is relatively motion-selective, whereas human V3 is less so. In macaque, the situation is qualitatively reversed: V3 is reported to be prominently motion-selective, whereas V3A is less so. As in human and macaque MT, the contrast sensitivity appears quite high in human areas V3 and V3A.


Evaluations of bone tissue integration to pure and alloyed titanium implants.
This study was performed for comparisons of tissue integration to commercially pure (CP) and titanium-6-aluminum-4 vanadium (Ti-6-Al-4V) implants using various existing three-dimensional biomechanical and two-dimensional histomorphometrical techniques, and to monitor the loosening torque during in vivo removal torque (RTQ) test with a novel unit not used before in a pilot study in rabbits. The implants were topographically characterized and inserted in femurs and tibiae of five rabbits (in total 40 implants, 20 per group). After 16 weeks, the implant integration was biomechanically evaluated by: (1) resonance frequency test, and (2) peak RTQ test and the graph from the monitoring curve. Biopsies of the implants in situ were processed to undecalcified cut and ground sections followed by light microscopical quantifications. Shear strength calculations were performed. Significantly higher mean value of RTQ (p = .01) and shear strength tests (p = .03) were observed for the CP titanium implants compared to Ti-6-Al-4V implants. The monitoring curve from the RTQ test demonstrated no differences in the shape or form that could provide further information about the differences in the implant-to-bone attachment. The CP titanium implants showed increased RTQ and shear strength values compared to the Ti-6-Al-4V implants. The new tool of monitoring the RTQ curve could not demonstrate differences between the two materials. The exact influence of the implant materials on the surrounding tissues needs to be further investigated.


Protofibrils, pores, fibrils, and neurodegeneration: separating the responsible protein aggregates from the innocent bystanders.
Many neurodegenerative diseases, including Alzheimer's and Parkinson's and the transmissible spongiform encephalopathies (prion diseases), are characterized at autopsy by neuronal loss and protein aggregates that are typically fibrillar. A convergence of evidence strongly suggests that protein aggregation is neurotoxic and not a product of cell death. However, the identity of the neurotoxic aggregate and the mechanism by which it disables and eventually kills a neuron are unknown. Both biophysical studies aimed at elucidating the precise mechanism of in vitro aggregation and animal modeling studies support the emerging notion that an ordered prefibrillar oligomer, or protofibril, may be responsible for cell death and that the fibrillar form that is typically observed at autopsy may actually be neuroprotective. A subpopulation of protofibrils may function as pathogenic amyloid pores. An analogous mechanism may explain the neurotoxicity of the prion protein; recent data demonstrates that the disease-associated, infectious form of the prion protein differs from the neurotoxic species. This review focuses on recent experimental studies aimed at identification and characterization of the neurotoxic protein aggregates.


Research on biological materials of human origin. Jurists and scientists face to face. Commentary.
On 3rd October 2016 a convention was held in the Aldo Moro room of the Chamber of Deputies on "Research on biological materials of human origin. Jurists and scientists face to face". The convention was organised by the Bioethics Unit of the Istituto Superiore di Sanità (ISS, Italian National Institute of Health) in conjunction with the Italian Academy of the Internet Code (IAIC) and the Fondazione Centro di Iniziativa Giuridica Piero Calamandrei. The present contribution reports the topics discussed and the key conclusions reached. As a follow-up to the discussion, the scientists, jurists and institutions concerned are resolved to take further steps towards the formulation of operational proposals intended to facilitate research using human biological materials within a framework of precise and strict regulations.


Variation in Brain Morphology of Intertidal Gobies: A Comparison of Methodologies Used to Quantitatively Assess Brain Volumes in Fish.
When correlating brain size and structure with behavioural and environmental characteristics, a range of techniques can be utilised. This study used gobiid fishes to quantitatively compare brain volumes obtained via three different methods; these included the commonly used techniques of histology and approximating brain volume to an idealised ellipsoid, and the recently established technique of X-ray micro-computed tomography (micro-CT). It was found that all three methods differed significantly from one another in their volume estimates for most brain lobes. The ellipsoid method was prone to over- or under-estimation of lobe size, histology caused shrinkage in the telencephalon, and although micro-CT methods generated the most reliable results, they were also the most expensive. Despite these differences, all methods depicted quantitatively similar relationships among the four different species for each brain lobe. Thus, all methods support the same conclusions that fishes inhabiting rock pool and sandy habitats have different patterns of brain organisation. In particular, fishes from spatially complex rock pool habitats were found to have larger telencephalons, while those from simple homogenous sandy shores had a larger optic tectum. Where possible we recommend that micro-CT be used in brain volume analyses, as it allows for measurements without destruction of the brain and fast identification and quantification of individual brain lobes, and minimises many of the biases resulting from the histology and ellipsoid methods.


Spectral and dosimetric characteristics of a D2O-moderated 252Cf calibration facility.
Calculations have been made for a D2O-moderated 252Cf assembly like that being used for the calibration of neutron dosimeters at the U.S. National Bureau of Standards and being proposed by the International Standards Organization. Leakage spectra at various distances from the assembly are given along with variations in dose-equivalent rate, average neutron energy and 235U/237Np fission ratio. The spectral shape changes rapidly near the spherical assembly and the dose-equivalent rate changes more rapidly than would be expected on the basis of the inverse-square dependence. Calibration of neutron dosimeters should therefore be made at distances greater than 15 cm from the surface. At large distances from the source, the dose equivalent per unit fluence for neutrons above 1 eV is 9.3 X 10(-9) rem cm2. The effects of the structural material, recent revisions to nuclear data files and changes in the spectrum of the source neutrons on the external field were investigated. These changes produce only about a 5% change in the neutron-dose equivalent rate. The structural material introduces negligible anisotropy in the radiation field.


The effect of 5A self-management program on medication adherence of epileptic patients during the COVID - 19 pandemic: A randomized controlled trial.
Adherence to medication adherence plays a vital role in controlling the problems and complications of epilepsy. During the COVID - 19 pandemic and limitations of face-to-face education, the use of distance education can play an important role in providing education to patients. The aim of this study is the effect of model 5A self-management training on medication adherence in epileptic patients. In this single-blind randomized controlled trial, 56 epileptic patients referred to Shiraz Namazi Hospital were divided into intervention and control groups using random allocation. Thereafter, 5A self - management training sessions were virtually held in 5 sessions in WhatsApp application for intervention group. The data collection tool was MMAS-8 at two stages of before and two months after intervention. Data analysis was performed using SPSS 21 software. The results of Wilcoxon test showed that in intervention group, the mean drug adherence in patients after intervention significantly increased compared to before intervention (p = 0.005). But in control group, this was not significant (p = 0.909). According to results of Mann-Whitney test, there was no significant difference between intervention and control groups before intervention (p = 0.632). However, by passing two months from intervention, a significant difference was found between intervention and control groups (p = 0.041). Based on results, the implementation of model 5A can be effective on medication adherence in epileptic patients. This program can be considered as a suitable method in epileptic patients during COVID - 19 pandemic.


[Effect of petroleum hydrocarbons on the viability of cyanobacteria in association with oil-oxidizing bacteria].
An important aspect in the problem of interactions between microorganisms in the conditions of oil pollution is how to preserve the viability of phototrophic organisms if active oil-oxidizing microflora is present in the environment. As was illustrated using a closed model ecosystem, the association 'cyanobacteria--oil-oxidizing bacteria' is capable of withstanding the negative effect of oil pollution, but within the range of hydrocarbon concentrations which can be oxidized by oil-oxidizing bacteria during a very short time. The biological equilibrium in the ecosystem was maintained and the number of viable cells of the phototrophic component in the ecosystem increased if the oil-oxidizing bacteria started to function at the same time as toxic compounds commenced to produce their effect on the microorganisms.


Persistence of micronuclei in lymphocytes of cancer patients after radiotherapy.
To verify the applicability of the micronucleus (MN) yield in peripheral blood lymphocytes (PBLs) as a quantitative biodosimeter for monitoring in vivo ionizing radiation damage, we applied the cytokinesis-blocked micronucleus assay in PBLs of cancer patients treated with partial-body radiotherapy. Dosimetric information on these 13 patients represented a wide range in the number of fractions, cumulative tumor dose, total integral dose, and equivalent total-body absorbed dose. We found in PBLs of these patients that (1) the MN yield increased linearly with the equivalent total-body absorbed dose (r = 0.8, P = 0.002), (2) the distributions of the MN yields deviated significantly from Poisson, and (3) there was a general decline in MN yields with increasing length of follow-up, but with considerable variation between individuals. The average rate of decline was found to be linear and was correlated with the equivalent total-body absorbed dose (r = 0.7, P = 0.007). Further, at 19-75 months of follow-up time, seven patients showed higher MN yields than their respective levels before radiotherapy, indicating the persistence of radiation-induced residual cytogenetic damage. Our findings suggest that the MN yield in human PBLs offers a reliable acute and perhaps chronic biodosimeter for in vivo radiation dose estimation. After the completion of radiotherapy, the persistence of elevated MN yield in PBLs is a reflection of the surviving population of radiation-induced genetically aberrant cells.


Assessment of oxygen transport and utilization in dogs with naturally occurring sepsis.
To compare cardiac index (CI), oxygen delivery index (D(O(2))I), oxygen extraction ratio (O(2)ER), oxygen consumption index (V(O(2))I), and systemic vascular resistance index (SVRI) in dogs with naturally occurring sepsis with published values for healthy dogs; compare those variables in dogs with sepsis that did or did not survive; and compare CI and D(O(2))I in dogs with sepsis with values in dogs with nonseptic systemic inflammatory response syndrome (nSIRS). Cohort study. 10 dogs with naturally occurring sepsis and 11 dogs with nSIRS. Over 24 hours, CI, D(O(2))I, O(2)ER, V(O(2))I, and SVRI were measured 4 and 5 times in dogs with sepsis and with nSIRS, respectively. The mean values of each variable in each group were compared over time and between groups; data for dogs with sepsis that did or did not survive were also compared. Mean D(O(2))I was significantly decreased, and mean CI, O(2)ER, V(O(2))I, and SVRI were not significantly different in dogs with sepsis, compared with published values for healthy dogs. Mean CI and D(O(2))I in dogs with sepsis were significantly greater than values in dogs with nSIRS. Among dogs with sepsis that did or did not survive, values of CI, D(O(2)I), O(2)ER, V(O(2))I, and SVRI did not differ significantly. Compared with values in healthy dogs, only D(O(2))I was significantly lower in dogs with sepsis. Values of CI and D(O(2))I were significantly higher in dogs with sepsis than in dogs with nSIRS, suggesting differing degrees of myocardial dysfunction between these groups.


Evidence for diminished visceral pain with aging: studies using graded intraesophageal balloon distension.
Graded intraesophageal balloon distension (IEBD) has been utilized in the past to evaluate esophageal pain thresholds. With use of a technique that we have found to provide reproducible results for pain thresholds, two groups of normal individuals without esophageal symptoms or diabetes were studied. Group 1 included 10 "young" (age < 65 yr) individuals (mean age 27 yr, range 18-57 yr). Group 2 included 17 individuals age 65 yr or greater (mean age 72.5 yr, range 65-87 yr). Catheters with latex balloons (Wilson-Cook) were used in all 27 subjects with the balloon located 10 cm above the lower esophageal sphincter. Sequential inflations of 2-ml increments were performed until a total volume of 2 ml above the point of pain or to a maximum of 30 ml was reached. A series of two sequential inflations were performed on each subject on the day of the testing, and the mean value was taken to indicate pain threshold volumes for all 27 subjects. In the group of elderly volunteers, 5 subjects felt no pain even at the maximum inflatable volume of the balloon (30 ml) and were assigned a maximum threshold value of 30 ml. Mean pain threshold volumes for the young subjects was 17 +/- 0.8 ml of air (+/- SE) and for the elderly subjects was 27 +/- 1.4 ml (P < 0.01 and 95% confidence interval = 7.1-13.3). Our conclusion is that IEBD results in the esophagus indicate an age-related decrease in human visceral pain threshold.


Microwave manipulation of an atomic electron in a classical orbit.
Although an atom is a manifestly quantum mechanical system, the electron in an atom can be made to move in a classical orbit almost indefinitely if it is exposed to a weak microwave field oscillating at its orbital frequency. The field effectively tethers the electron, phase-locking its motion to the oscillating microwave field. By exploiting this phase-locking, we have sped up or slowed down the orbital motion of the electron in excited lithium atoms by increasing or decreasing the microwave frequency between 13 and 19 gigahertz; the binding energy and orbital size change concurrently.


Progressive hyperplastic lesions of the bladder uroepithelium after hormone stimulation in Nb rats.
The bladder epithelium of female Nb rats was markedly stimulated by treatment with pellets of testosterone propionate. Estrone treatment simultaneously reduced the incidence of bladder changes to approximately 50 per cent although estrone alone had no effect on bladder weights. The bladders of male rats were less responsive to hormones and were stimulated only by treatment with estrogen + testosterone. Massive papillary hyperplasia of the uroepithelium that extended into the kidney pelvis could be produced by hormones in the absence of calculi. Transplants of hyperplastic bladder epithelium usually resulted in small pseudo-bladders, but in one case a transplantable squamous cell carcinoma developed.


[The effect of elevated ozone levels in the ambient air on lung function of children engaged in sports].
In the summer of 1989, ozone levels in the ambient air were regularly found to be increased in The Netherlands. The question was studied whether this affected pulmonary function of children aged 7 to 12 years practising sports in the open air. Before and after sessions of training in the open air, the peak expiratory flow rate was determined using peak flow meters (mini-Wright). This was done on 37 different days with varying ozone concentrations. Measurements were carried out on four or more days in a total of 65 children. No correlation between ozone concentration during training and differences between pre- and post-training peak flows was found. No correlation was found either between the maximal ozone concentration on a given day and the peak flow at the end of training on that day. However, a negative correlation was found between the maximal ozone concentration on the day preceding the pulmonary function tests and the peak flow after the training.


Accuracy of advanced versus strictly conventional 12-lead ECG for detection and screening of coronary artery disease, left ventricular hypertrophy and left ventricular systolic dysfunction.
Resting conventional 12-lead ECG has low sensitivity for detection of coronary artery disease (CAD) and left ventricular hypertrophy (LVH) and low positive predictive value (PPV) for prediction of left ventricular systolic dysfunction (LVSD). We hypothesized that a approximately 5-min resting 12-lead advanced ECG test ("A-ECG") that combined results from both the advanced and conventional ECG could more accurately screen for these conditions than strictly conventional ECG. Results from nearly every conventional and advanced resting ECG parameter known from the literature to have diagnostic or predictive value were first retrospectively evaluated in 418 healthy controls and 290 patients with imaging-proven CAD, LVH and/or LVSD. Each ECG parameter was examined for potential inclusion within multi-parameter A-ECG scores derived from multivariate regression models that were designed to optimally screen for disease in general or LVSD in particular. The performance of the best retrospectively-validated A-ECG scores was then compared against that of optimized pooled criteria from the strictly conventional ECG in a test set of 315 additional individuals. Compared to optimized pooled criteria from the strictly conventional ECG, a 7-parameter A-ECG score validated in the training set increased the sensitivity of resting ECG for identifying disease in the test set from 78% (72-84%) to 92% (88-96%) (P < 0.0001) while also increasing specificity from 85% (77-91%) to 94% (88-98%) (P < 0.05). In diseased patients, another 5-parameter A-ECG score increased the PPV of ECG for LVSD from 53% (41-65%) to 92% (78-98%) (P < 0.0001) without compromising related negative predictive value. Resting 12-lead A-ECG scoring is more accurate than strictly conventional ECG in screening for CAD, LVH and LVSD.


Adaptive postprocessing algorithms for low bit rate video signals.
A new adaptive postprocessing algorithm to enhance the quality of a noisy video sequence is presented. The algorithm recognizes that the visibility of noise depends on local signal characteristics. It therefore classifies the video signal into different classes and uses separate nonlinear filters matched to each class. The most general version of the algorithm employs motion-compensated frame averaging to improve picture quality in a first stage. A classification algorithm subsequently divides subblocks of pixels in the averaged frame into four classes: edge, smooth, nonsmooth with motion and nonsmooth without motion. Spatial algorithms that perform multilevel median filtering, double median filtering, and median filtering are used for pixels belonging to edge, smooth, and nonsmooth with motion categories. Pixels in the nonsmooth, unmoving category are left unfiltered to preserve corresponding image texture. In a simpler version of this four-class system, the motion cues and motion-compensated frame averaging are eliminated, and the purely spatial filtering is based on a three-class algorithm. When used at the output of a 3-D subband coder at 384 kbps, the spatial postfilter was shown to provide a consistent gain in subjectively evaluated picture quality. Twenty-five viewers participated in an experiment involving three coded sequences. In a pairwise comparison of postfiltered and unfiltered sequences, the postfiltered version was judged to be better in 63 out of 75 instances.


Two-dimensional Cu2Si monolayer with planar hexacoordinate copper and silicon bonding.
Two-dimensional (2D) materials with planar hypercoordinate motifs are extremely rare due to the difficulty in stabilizing the planar hypercoordinate configurations in extended systems. Furthermore, such exotic motifs are often unstable. We predict a novel Cu2Si 2D monolayer featuring planar hexacoordinate copper and planar hexacoordinate silicon. This is a global minimum in 2D space which displays reduced dimensionality and rule-breaking chemical bonding. This system has been studied with density functional theory, including molecular dynamics simulations and electronic structure calculations. Bond order analysis and partitioning reveals 4c-2e σ bonds that stabilize the two-dimensional structure. We find that the system is quite stable during short annealing simulations up to 900 K, and predict that it is a nonmagnetic metal. This work opens up a new branch of hypercoordinate two-dimensional materials for study.


[Association between insomnia and rheumatoid arthritis in elderly].
This study aimed to assess symptoms of insomnia in elderly residents in the community and its association with rheumatoid arthritis. Descriptive and cross-sectional study, part of a multicentre research project entitled Fragility in Brazilian elderly . A total number of 689 elderly (68.9% female, average age of 72.2 years) were interviewed using a questionnaire on sociodemographic data and clinical conditions. Two tools to identify symptoms of insomnia (the Nottingham Health profile) and to screen cognitive impairment (Mini Mental State Examination) were also applied. For data analysis, we used descriptive statistical techniques and multiple logistic regression, considering a 5% significance level. The association between insomnia symptoms and rheumatoid arthritis was found to be significant in the multivariate analysis; other associated factors were a very poor subjective evaluation of health, being retired, and the presence of depression. Health professionals should investigate carefully the sleep quality in elderly with rheumatoid arthritis, considering its high prevalence in this population.


MIB-1 Is Required for Spermatogenesis and Facilitates LIN-12 and GLP-1 Activity in Caenorhabditis elegans.
Covalent attachment of ubiquitin to substrate proteins changes their function or marks them for proteolysis, and the specificity of ubiquitin attachment is mediated by the numerous E3 ligases encoded by animals. Mind Bomb is an essential E3 ligase during Notch pathway signaling in insects and vertebrates. While Caenorhabditis elegans encodes a Mind Bomb homolog (mib-1), it has never been recovered in the extensive Notch suppressor/enhancer screens that have identified numerous pathway components. Here, we show that C. elegans mib-1 null mutants have a spermatogenesis-defective phenotype that results in a heterogeneous mixture of arrested spermatocytes, defective spermatids, and motility-impaired spermatozoa. mib-1 mutants also have chromosome segregation defects during meiosis, molecular null mutants are intrinsically temperature-sensitive, and many mib-1 spermatids contain large amounts of tubulin. These phenotypic features are similar to the endogenous RNA intereference (RNAi) mutants, but mib-1 mutants do not affect RNAi. MIB-1 protein is expressed throughout the germ line with peak expression in spermatocytes followed by segregation into the residual body during spermatid formation. C. elegans mib-1 expression, while upregulated during spermatogenesis, also occurs somatically, including in vulva precursor cells. Here, we show that mib-1 mutants suppress both lin-12 and glp-1 (C. elegans Notch) gain-of-function mutants, restoring anchor cell formation and a functional vulva to the former and partly restoring oocyte production to the latter. However, suppressed hermaphrodites are only observed when grown at 25°, and they are self-sterile. This probably explains why mib-1 was not previously recovered as a Notch pathway component in suppressor/enhancer selection experiments.


Permeability of endothelial monolayers to albumin is increased by bradykinin and inhibited by prostaglandins.
Using monolayers of bovine aortic endothelial cells (BAEC) in modified Boyden chambers, we examined the role of prostaglandins (PGs) in the bradykinin (BK)-induced increase of albumin permeability. BK induced a concentration-dependent increase of the permeability of BAEC, which reached 49.9 +/- 1% at the concentration of 10(-8) M. Two inhibitors of the prostaglandin G/H synthase, indomethacin (2.88 microM) and ibuprofen (10 microM), potentiated BK-induced permeability 1.8- and 3.9-fold, respectively. Exogenously administered PGE2 and iloprost, a stable analog of prostacyclin, attenuated the effect of BK in a concentration-dependent manner. Butaprost equally reduced the effect of BK, suggesting the participation of the EP2 receptor in this phenomenon. However, the EP4-selective antagonist AH-23848 did not significantly inhibit the protective effect of PGE2. The inhibitory effect of PGE2 was reversed by the adenylate cyclase inhibitor MDL-12330A (10 microM). These results suggest that BK-induced increase of permeability of BAEC monolayer to (125)I-labeled albumin is negatively regulated by PGs. This postulated autocrine activity of PGs may involve an increase in the intracellular level of cAMP.


Ploidy evolution in a wild yeast is linked to an interaction between cell type and metabolism.
Ploidy is an evolutionarily labile trait, and its variation across the tree of life has profound impacts on evolutionary trajectories and life histories. The immediate consequences and molecular causes of ploidy variation on organismal fitness are frequently less clear, although extreme mating type skews in some fungi hint at links between cell type and adaptive traits. Here, we report an unusual recurrent ploidy reduction in replicate populations of the budding yeast Saccharomyces eubayanus experimentally evolved for improvement of a key metabolic trait, the ability to use maltose as a carbon source. We find that haploids have a substantial, but conditional, fitness advantage in the absence of other genetic variation. Using engineered genotypes that decouple the effects of ploidy and cell type, we show that increased fitness is primarily due to the distinct transcriptional program deployed by haploid-like cell types, with a significant but smaller contribution from absolute ploidy. The link between cell-type specification and the carbon metabolism adaptation can be traced to the noncanonical regulation of a maltose transporter by a haploid-specific gene. This study provides novel mechanistic insight into the molecular basis of an environment-cell type fitness interaction and illustrates how selection on traits unexpectedly linked to ploidy states or cell types can drive karyotypic evolution in fungi.


Effects of acute adult and early-in-life bladder inflammation on bladder neuropeptides in adult female rats.
The purpose of the present study was to determine how acute adult and/or prior early-in life (EIL; P14-P16) exposure to bladder inflammation affects bladder content of calcitonin gene related peptide (CGRP) and substance P (SP). Estrous cycle influences were also studied in the adult-treatment conditions. In Experiment 1, intravesical zymosan or isoflurane anesthesia alone was administered to adult female rats. Bladders and serum were collected 24 hours later during each phase of the estrous cycle. In Experiment 2, zymosan or anesthesia alone was administered EIL and as adults, with bladder tissue collection 24 h later. In general, Experiment 1 showed that bladder content of both CGRP and SP was increased by inflammation. This effect was significant when data were collapsed across all phases of the estrous cycle, but was only significant during proestrus when individual comparisons were made during each phase of estrous. Also, adult bladder inflammation significantly reduced estradiol levels. In Experiment 2, bladder content of CGRP and SP was significantly increased in rats receiving EIL and/or adult inflammation. Bladder weights were also significantly increased by inflammation. These data indicate that bladder CGRP and SP are maximally increased during the proestrus phase of the estrous cycle in inflamed adult female rats. EIL exposure to bladder inflammation alone can also produce an increase in CGRP and SP lasting into adulthood. Therefore, EIL experience with bladder inflammation may predispose an organism to experience a painful bladder disorder as an adult by increasing primary afferent content of CGRP and/or SP.


Drug removal during continuous arteriovenous hemofiltration: theory and clinical observations.
We have discussed the basic principles of pharmacokinetics and convective solute removal in the context of each other. Clinical observations appear to follow the theoretical expectations. For practical purposes plasma and plasma water are not different. In the calculation of drug sieving, venous samples do not contribute enough to warrant their extra costs. We recommend that drug removal in hemofiltration be expressed by the sieving coefficient, UF/A. Drug sieving data in humans undergoing CAVH are tabulated. Recommendations for supplemental dosing are discussed which are applicable to any clinical setting.


Protective effect of liver ischemic preconditioning on liver and lung injury induced by hepatic ischemia-reperfusion in the rat.
This study evaluates whether preconditioning could modulate the injurious effects of tumor necrosis factor (TNF) on liver and lung following hepatic ischemia-reperfusion (I/R) by inhibiting hepatic postischemic TNF release. The inhibition of hepatic TNF release from Kupffer cells with gadolinium chloride (GdCl(3)) previous to ischemia maintained TNF at control levels, attenuating the increases in transaminases, vascular permeability, and edema associated with hepatic I/R injury. TNF addition reverted this beneficial effect, indicating the implication of the TNF released mainly from Kupffer cells in hepatic I/R injury. Preconditioning prevented hepatic TNF increases, thus attenuating the liver injury, while TNF addition abolished the benefits of preconditioning. Inhibition of nitric oxide (NO) synthesis abolished the effect of preconditioning, whereas GdCl(3) addition avoided the injurious effect of NO inhibition. In addition, NO administration before I/R offered similar results to those found in preconditioning, while TNF addition abolished the benefits of NO. Thus, the effect of preconditioning on TNF release after hepatic I/R is mediated by NO. Inhibition of hepatic TNF release from Kupffer cells with GdCl(3) prevented both the increase in plasma TNF and the injurious effect in lung seen after hepatic I/R, and these effects were reverted with TNF addition. Preconditioning resulting in reduced hepatic TNF levels prevented the systemic TNF release, thus reducing the lung damage following hepatic I/R. However, TNF addition abolished the protective effect of preconditioning on lung injury. These findings indicate that preconditioning attenuates hepatic postischemic TNF release from Kupffer cells, thus probably reducing the liver and lung injury following hepatic I/R, and that this effect of preconditioning is mediated by NO.


Single-molecule biomechanics with optical methods.
Single-molecule observation and manipulation have come of age. With the advent of optical tweezers and other methods for probing and imaging single molecules, investigators have circumvented the model-dependent extrapolation from ensemble assays that has been the hallmark of classical biochemistry and biophysics. In recent years, there have been important advances in the understanding of how motor proteins work. The range of these technologies has also started to expand into areas such as DNA transcription and protein folding. Here, recent experiments with rotary motors, linear motors, RNA polymerase, and titin are described.


Biological Assembly of Modular Protein Building Blocks as Sensing, Delivery, and Therapeutic Agents.
Nature has evolved a wide range of strategies to create self-assembled protein nanostructures with structurally defined architectures that serve a myriad of highly specialized biological functions. With the advent of biological tools for site-specific protein modifications and de novo protein design, a wide range of customized protein nanocarriers have been created using both natural and synthetic biological building blocks to mimic these native designs for targeted biomedical applications. In this review, different design frameworks and synthetic decoration strategies for achieving these functional protein nanostructures are summarized. Key attributes of these designer protein nanostructures, their unique functions, and their impact on biosensing and therapeutic applications are discussed.


The Optimal Outcome of Suppressing Ewing Sarcoma Growth in vivo With Biocompatible Bioengineered miR-34a-5p Prodrug.
Being the second most common type of primary bone malignancy in children and adolescents, Ewing Sarcoma (ES) encounters the dilemma of low survival rate with a lack of effective treatments. As an emerging approach to combat cancer, RNA therapeutics may expand the range of druggable targets. Since the genome-derived oncolytic microRNA-34a (miR-34a) is down-regulated in ES, restoration of miR-34a-5p expression or function represents a new therapeutic strategy which is, however, limited to the use of chemically-engineered miRNA mimics. Very recently we have developed a novel bioengineering technology using a stable non-coding RNA carrier (nCAR) to achieve high-yield production of biocompatible miRNA prodrugs, which is a great addition to current tools for the assessment of RNA therapeutics. Herein, for the first time, we investigated the biochemical pharmacology of bioengineered miR-34a-5p prodrug (nCAR/miR-34a-5p) in the control of ES using human ES cells and xenograft mouse models. The bioengineered nCAR/miR-34a-5p was precisely processed to mature miR-34a-5p in ES cells and subsequently suppressed cell proliferation, attributable to the enhancement of apoptosis and induction of G2 cell cycle arrest through downregulation of SIRT-1, BCL-2 and CDK6 protein levels. Furthermore, systemic administration of nCAR/miR-34a-5p dramatically suppressed the ES xenograft tumor growth in vivo while showing biocompatibility. In addition, the antitumor effect of bioengineered nCAR/miR-34a-5p was associated with a lower degree of tumoral cell proliferation and greater extent of apoptosis. These findings demonstrate the efficacy of bioengineered miR-34a-5p prodrug for the treatment of ES and support the development of miRNA therapeutics using biocompatible bioengineered miRNA prodrugs.


Calmodulin-dependent enzymic phosphorylation of dolichol in Tetrahymena pyriformis.
The Protozoan, Tetrahymena pyriformis, is capable of phosphorylating dolichol in the presence of CTP. Other nucleotides (ATP, UTP and GTP) were ineffective. The enzyme was activated independently by the bivalent cations Mg2+, Mn2+ and Ca2+. The Ca2+ stimulation of the enzyme activity was calmodulin-dependent. A substantial increase in the enzyme activity was seen in the presence of UTP. The apparent Km values for CTP and dolichol, as calculated from the Lineweaver-Burk plot, were 3 mM and 70 microM respectively. Although the presence of detergent was essential, at higher concentrations there was a decrease in the enzyme activity. The enzyme had two pH optima (pH 7.4 and 9.0) and at both the activity was Ca2+-calmodulin-dependent.


Human cerebral circuitry related to cardiac control: A neuroimaging meta-analysis.
Serious arrhythmias often follow stroke, highlighting the important role that the cerebral cortex has in neural regulation of the heart. Neuroimaging studies reveal patterns of activation associated with autonomic cardiovascular responses. Using the activation likelihood estimation, a meta-analysis was performed to identify cortical activation patterns related to heart rate and heart rate variability across studies that used differing stimuli of cardiovascular arousal. Consistent associations with heart rate and heart rate variability were observed in the anterior cingulate, amygdala, insula, and prefrontal cortex, among others. Also, evidence is provided of insular lateralization associated with autonomic cardiac responses. Ann Neurol 2016;79:709-716.


Leber hereditary optic neuropathy: current perspectives.
Leber hereditary optic neuropathy (LHON) is one of the most common inherited optic neuropathies causing bilateral central vision loss. The disorder results from point mutations in mitochondrial DNA and subsequent mitochondrial dysfunction. The primary cell type that is lost in LHON is the retinal ganglion cell, which is highly susceptible to disrupted ATP production and oxidative stress. Inheritance of LHON follows that of mitochondrial genetics, and it has a highly variable clinical phenotype, as other genetic and environmental factors also play a role. Although LHON usually presents with isolated vision loss, some patients suffer other neurological sequelae. For ill-defined reasons, male LHON mutation carriers are more affected than females. Most LHON patients remain legally blind, but a small proportion can experience spontaneous partial recovery, often within the first year of symptom onset. Unfortunately, at this time there are no established curative interventions and treatment is largely supportive. Patients should be offered low vision services and counseled on mitigating risk factors for additional vision loss, such as smoking and consuming alcohol. Encouraging treatments currently undergoing investigation includes ubiquinone analogs, such as idebenone, as well as gene therapy and stem cells to restore ATP synthesis and provide neuroprotection to surviving retinal ganglion cells.


Fitting of bone mineral density with consideration of anthropometric parameters.
A new model describing normal values of bone mineral density in children has been evaluated, which includes not only the traditional parameters of age, gender, and race, but also weight, height, percent body fat, and sexual maturity. This model may constitute a better comparative norm for a specific child with given anthropometric values. Previous descriptions of children's bone mineral density (BMD) by age have focused on segmenting diverse populations by race and gender without adjusting for anthropometric variables or have included the effects of anthropometric variables over a relatively homogeneous population. Multivariate semi-metric smoothing (MS(2)) provides a way to describe a diverse population using a model that includes multiple effects and their interactions while producing a result that can be smoothed with respect to age in order to provide connected percentiles. We applied MS(2) to spine BMD data from the Bone Mineral Density in Childhood Study to evaluate which of gender, race, age, height, weight, percent body fat, and sexual maturity explain variations in the population's BMD values. By balancing high adjusted R (2) values and low mean square errors with clinical needs, a model using age, gender, race, weight, and percent body fat is proposed and examined. This model provides narrower distributions and slight shifts of BMD values compared to the traditional model, which includes only age, gender, and race. Thus, the proposed model might constitute a better comparative standard for a specific child with given anthropometric values and should be less dependent on the anthropometric characteristics of the cohort used to devise the model. The inclusion of multiple explanatory variables in the model, while creating smooth output curves, makes the MS(2) method attractive in modeling practically sized data sets. The clinical use of this model by the bone research community has yet to be fully established.