J4 at sweet 16: a new wrinkle?
Compartmental models provide a major source of insight into the information processing functions of single neurons. Over the past 15 years, one of the most widely used neuronal morphologies has been the cell called "j4," a layer 5 pyramidal cell from cat visual cortex originally described in Douglas, Martin, and Whitteridge (1991). The cell has since appeared in at least 28 published compartmental modeling studies, including several in this journal. In recently examining why we could not reproduce certain in vitro data involving the attenuation of signals originating in distal basal dendrites, we discovered that pronounced fluctuations in the diameter measurements of j4 lead to a bottlenecking effect that increases distal input resistances and significantly reduces voltage transfer between distal sites and the cell body. Upon smoothing these diameter fluctuations, bringing j4 more in line with other reconstructions of layer 5 pyramidal neurons, we found that the attenuation of steady-state voltage signals traveling to the cell body V(distal)/V(soma) was reduced by 60% at some locations in some branches (corresponding to a 2.5-fold increase in the voltage response at the soma for the same distal depolarization) and by 30% on average (corresponding to a 45% increase in somatic response). Changes of this magnitude could lead to different outcomes in some types of compartmental modeling studies. A smoothed version of the j4 morphology is available online at http://lnc.usc.edu/j4-smooth/ .


[INFLUENCE OF THE NUTRITIONAL COMPOSITION OF DIFFERENT FIBER-ENRICHED ENTERAL NUTRITION FORMULAS ON THE ADMINISTRATION TIME BY GRAVITY AND THE RISK OF TUBE FEEDING OBSTRUCTION].
the administration of enteral nutrition by gravity is a very useful method in clinical practice; nevertheless, it may not be very precise. Indeed, this method presents some important limitations, such as the difficulty in establishing a precise dripping rate and the possibility for the dripping rate decrease depending on the formula. assess the administration time and the risk of clogging of 5 fiber-enriched enteral nutrition formulas with different protein concentrations and caloric density, all administered by gravity through nasogastric (NG) tubes of different sizes. Assess the influence of the composition on the dripping rate, by gravity, of the tested formulas. 5 fiber-enriched EN formulas were compared by using nasogastric tubes of the calibers 8, 10 and 12 Fr. The fluidity of these gravity-administered NE formulas was estimated by timing the complete passage of each formula at full speed, thus allowing one to calculate the mean time of free fall (MTFF) and to register any possible obstruction. Subsequently, an in vitro simulation of a 1 500 ml administration was performed for each formula at a particular speed, so that the administration time was 5 hours. Slowing flow and stagnated flow were detected as indicators of the risk of obstruction. the two products that especially differed in MTFF were the ones with the highest energy concentration. The passage time in free fall of these two products through the 8 Fr tube exceeded four hours. For the rest of the products and NG tubes used, this time was less than 2 hours and 5 minutes. No slowing flow or tube obstruction was detected in free fall and at maximum speed. When the dripping was adjusted to be administered in 5 hours, three of the studied products (those with the least caloric concentration and viscosity) showed slowing flow and, in some cases, the dripping stopped completely. The most important factor associated to the MTFF was the lipid content, followed by viscosity, energy and protein content. The MTFF measured was not significantly related to the fiber content of the nutritional formula. all studied products can be administered by gravity via nasogastric tubes in free fall without any risk of obstruction, even though the free fall time was very variable. The lowest caliber tubes, the highest energy content and the viscosity of the EN mixture turn-out to be the limiting factors when fiber-enriched formulas are to be administered by gravity.


Impact of Superoxide Dismutase-Gliadin on Radiation-induced Fibrosis: An Experimental Study.
Radiation-induced fibrosis (RIF) has since long been considered as irreversible. Further understanding of its mechanisms has led to trials investigating RIF treatment and prevention. The effect of superoxide dismutase (SOD)-gliadin, an oral form of SOD that resists gastrointestinal inactivation, on RIF treatment was evaluated in this experimental study. A total of 36 Wistar albino mice were randomly distributed into four groups. According to group, 25 Gy radiation or sham-radiation were performed on day 0. Acute and late reactions were recorded. After 6 months, mice were treated with SOD-gliadin, 10,000 units per kg per day, or placebo. SOD-gliadin and placebo treatments were administered daily for 8 days by oral gavage. Later the mice were sacrificed, dissected and histopathologically analyzed. Accumulated hyaline and collagen at the dermis is an indicator of fibrosis. Therefore measurements of the dermal thickness were used to quantify the degree of RIF. Additionally, the morphological changes were analyzed, and the differences reported. The mean and standard deviation for dermal thickness were 0.45±0.09 mm in the sham-irradiated placebo-treated group, 0.51 mm±0.16 mm in the sham-irradiated SOD-gliadin-treated group, 0.92 mm±0.23 mm in the irradiated placebo-treated group and 0.71 mm±0.17 mm in the irradiated SOD-gliadin-treated group. The difference in mean dermal thickness between irradiated placebo-treated and irradiated SOD-gliadin-treated mice was statistically significant (p=0.002). Quality of life while prolonging survival has an increasing importance in patients with cancer. RIF can be a crucial problem after all radiotherapy modalities. SOD-gliadin has advantageous effects on conditions that call for an increased expression of antioxidant enzymes. The results of our study suggest that oral SOD-gliadin may prevent or ameliorate RIF and patients can benefit from the positive effects of SOD.


Adverse perinatal outcome in patients screen-positive for neural tube defects and fetal Down syndrome.
An association between various abnormal mid-trimester maternal serum analyte values and adverse perinatal outcome has been reported. From an original sample of 14,857 women, we observed five women who were 'screen-positive' for both neural tube defects [maternal serum alpha-fetoprotein (MSAFP) > or = 2.5 multiples of the median] and Down syndrome [risk > or = 1/274 using MSAFP, maternal serum unconjugated oestriol (MSuE3), maternal serum human chorionic gonadotropin (MShCG), and maternal age]. The four patients who elected to undergo amniocentesis all demonstrated both normal karyotype and normal amniotic fluid AFP levels. All five cases were associated with intrauterine growth retardation (IUGR) and abnormal pregnancy outcomes. Two cases exhibiting severe IUGR on ultrasound examination were terminated at 19.1 and 21.2 weeks, respectively; the former also exhibited fetal calcifications and positive maternal serology for toxoplasmosis. In another case, fetal demise occurred at 36 weeks' gestation in a patient who had been treated for syphilis in the second trimester. Neither infection was confirmed in fetal tissue studies. Though resulting in live births, the remaining two cases required operative deliveries; emergency Caesarean sections for fetal distress were performed at 38 and 32 weeks, respectively, the latter case being associated with severe pre-eclampsia. We conclude that elevated mid-trimester MSAFP levels concurrent with maternal serum analyte values associated with increased risk for fetal Down syndrome may presage a poor perinatal outcome, particularly IUGR and possibly congenital infection.


Myocardial infarction in Mexican-Americans and non-Hispanic whites. The San Antonio Heart Study.
Mexican-American men experience reduced cardiovascular mortality compared with non-Hispanic white men. There is no corresponding ethnic difference in cardiovascular mortality in women. The difference in men could result either from a lower incidence of cardiovascular disease or a lower case fatality rate among Mexican-Americans. Although the incidence of cardiovascular disease in Mexican-Americans is unknown, we have collected data on prevalence of myocardial infarction in 5,148 individuals examined in the San Antonio Heart Study, a population-based survey of cardiovascular disease conducted between 1979 and 1988 in Mexican-Americans and non-Hispanic whites aged 25-64 years. Myocardial infarction was assessed by Minnesota-coded electrocardiograms and by a self-reported history of a physician-diagnosed heart attack. For both end points, the age-adjusted prevalence of myocardial infarction was lower in Mexican-American men than in non-Hispanic white men. After adjustment for age and diabetes status (present/absent), the odds of a myocardial infarction, as defined by either criterion, was approximately one third lower in Mexican-American men than in non-Hispanic white men (p = 0.06). In women, the prevalence of both myocardial infarction end points was slightly higher in Mexican-Americans than in non-Hispanic whites, although neither of these differences was significant. Although the ethnic differences in prevalence in this study were not statistically significant, their pattern parallels the pattern in the mortality due to cardiovascular diseases. Therefore, the results support the hypothesis that the reduced cardiovascular mortality rate observed in Mexican-American men reflects a lower incidence of myocardial infarction rather than a reduced case fatality rate because the latter would result in a higher prevalence.


Usability and acceptability of an automated respiratory rate counter to assess childhood pneumonia in Nepal.
Pneumonia is the leading cause of child death after the neonatal period, resulting from late care seeking and inappropriate treatment. Diagnosis involves counting respiratory rate (RR); however, RR counting remains challenging for health workers and miscounting, and misclassification of RR is common. We evaluated the usability of a new automated RR counter, the Philips Children's Respiratory Monitor (ChARM), to Female Community Health Volunteers (FCHVs), and its acceptability to FCHVs and caregivers in Nepal. A cross-sectional study was conducted in Jumla district, Nepal. About 133 FCHVs were observed between September and December 2018 when using ChARM during 517 sick child consultations, 264 after training and 253 after 2 months of routine use of ChARM. Acceptability of the ChARM was explored using semi-structured interviews. FCHV adherence to guidelines after 2 months of using ChARM routinely was 52.8% (95% CI 46.6-58.9). The qualitative findings suggest that ChARM is acceptable to FCHVs and caregivers; however, capacity constraints such as older age and low literacy and impacted device usability were mentioned. Further research on the performance, cost-effectiveness and implementation feasibility of this device is recommended, especially among low-literate CHWs.


Population Diversity within Isolates of Colletotrichum spp. Causing Glomerella Leaf Spot and Bitter Rot of Apples in Three Orchards in North Carolina.
The population diversity within isolates of Glomerella cingulata and Colletotrichum spp. associated with Glomerella leaf spot and bitter rot of apples was studied in an orchard of cv. Granny Smith located in Wilkes County, NC, and one orchard each of cultivars Granny Smith and Gala located in Lincoln County, NC. Morphological characters and vegetative compatibility groups (VCGs) were used to determine diversity within the species. The relative frequencies of the morphological types found within each species in each orchard were also determined. G. cingulata was the predominant species associated with bitter rot in the three orchards and Glomerella leaf spot in the Gala orchard. In the three orchards, different morphological types were observed within isolates of G. cingulata and Colletotrichum acutatum, but not within isolates of Colletotrichum gloeosporioides. Isolates of C. gloeosporioides were not found in the orchard of cv. Granny Smith in Lincoln County. In the other two orchards, C. gloeosporioides represented the lowest proportion of the population. Three VCGs were found among isolates of G. cingulata (VCG-1, 2, and 6), two among isolates of C. gloeosporioides (VCG-9 and 10), and two among isolates of C. acutatum (VCG-15 and 16). VCGs 2, 6, 9, 10, 15, and 16 were found in the Granny Smith orchard in Wilkes County, VCGs 1, 2, and 6 in the Gala orchard in Lincoln County, and VCGs 2 and 6 in the Granny Smith orchard in Lincoln County. Differences in frequencies among the different morphological types found within the three orchards remained relatively similar throughout the season and from year to year, suggesting that the relative frequencies of G. cingulata, C. gloeosporioides, and C. acutatum remain stable in an orchard once the fungi are established.


Altered cardiovascular reactivity and osmoregulation during hyperosmotic stress in adult rats developmentally exposed to polybrominated diphenyl ethers (PBDEs).
Polybrominated diphenyl ethers (PBDEs) and the structurally similar chemicals polychlorinated biphenyls (PCBs) disrupt the function of multiple endocrine systems. PCBs and PBDEs disrupt the secretion of vasopressin (VP) from the hypothalamus during osmotic activation. Since the peripheral and central vasopressinergic axes are critical for osmotic and cardiovascular regulation, we examined whether perinatal PBDE exposure could impact these functions during physiological activation. Rats were perinatally dosed with a commercial PBDE mixture, DE-71. Dams were given 0 (corn oil control), 1.7 (low dose) or 30.6 mg/kg/day (high dose) in corn oil from gestational day (GD) 6 through postnatal day (PND) 21 by oral gavage. In the male offspring exposed to high dose PBDE plasma thyroxine and triiodothyronine levels were reduced at PND 21 and recovered to control levels by PND 60 when thyroid stimulating hormone levels were elevated. At 14-18 months of age, cardiovascular responses were measured in four groups of rats: Normal (Oil, normosmotic condition), Hyper (Oil, hyperosmotic stress), Hyper PBDE low (1.7 mg/kg/day DE-71 perinatally, hyperosmotic stress), and Hyper PBDE high (30.6 mg/kg/day DE-71 perinatally, hyperosmotic stress). Systolic blood pressure (BP), diastolic BP, and heart rate (HR) were determined using tail cuff sphygmomanometry and normalized to pretreatment values (baseline) measured under basal conditions. Hyperosmotic treatment yielded significant changes in systolic BP in PBDE exposed rats only. Hyper PBDE low and high dose rats showed 36.1 and 64.7% greater systolic BP responses at 3h post hyperosmotic injection relative to pretreatment baseline, respectively. No treatment effects were measured for diastolic BP and HR. Hyper and Hyper PBDE rats showed increased mean plasma osmolality values by 45 min after injection relative to normosmotic controls. In contrast to Hyper rats, Hyper PBDE (high) rats showed a further increase in mean plasma osmolality at 3h (358.3±12.4mOsm/L) relative to 45 min post hyperosmotic injection (325.1±11.4mOsm/L). Impaired osmoregulation in PBDE-treated animals could not be attributed to decreased levels of plasma vasopressin. Our findings suggest that developmental exposure to PBDEs may disrupt cardiovascular reactivity and osmoregulatory responses to physiological activation in late adulthood.


Isoprenylation is required for the processing of the lamin A precursor.
The nuclear lamina proteins, prelamin A, lamin B, and a 70-kD lamina-associated protein, are posttranslationally modified by a metabolite derived from mevalonate. This modification can be inhibited by treatment with (3-R,S)-3-fluoromevalonate, demonstrating that it is isoprenoid in nature. We have examined the association between isoprenoid metabolism and processing of the lamin A precursor in human and hamster cells. Inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase by mevinolin (lovastatin) specifically depletes endogenous isoprenoid pools and inhibits the conversion of prelamin A to lamin A. Prelamin A processing is also blocked by mevalonate starvation of Mev-1, a CHO cell line auxotrophic for mevalonate. Moreover, inhibition of prelamin A processing by mevinolin treatment is rapidly reversed by the addition of exogenous mevalonate. Processing of prelamin A is, therefore, dependent on isoprenoid metabolism. Analysis of the conversion of prelamin A to lamin A by two independent methods, immunoprecipitation and two-dimensional nonequilibrium pH gel electrophoresis, demonstrates that a precursor-product relationship exists between prelamin A and lamin A. Analysis of R,S-[5-3H(N)]mevalonate-labeled cells shows that the rate of turnover of the isoprenoid group from prelamin A is comparable to the rate of conversion of prelamin A to lamin A. These results suggest that during the proteolytic maturation of prelamin A, the isoprenylated moiety is lost. A significant difference between prelamin A processing, and that of p21ras and the B-type lamins that undergo isoprenylation-dependent proteolytic maturation, is that the mature form of lamin A is no longer isoprenylated.


The role of Polycomb in stem cell genome architecture.
Polycomb-group proteins maintain embryonic stem cell identity by repressing genes that encode for developmental regulatory factors. Failure to properly control developmental transcription programs by Polycomb proteins is linked to disease and embryonic lethality. Recent technological advances have revealed that developmentally repressed genes tend to cluster in the three-dimensional space of the nucleus. Importantly, spatial clustering of developmental genes is fundamental for the correct regulation of gene expression during early development. Here, we outline novel insights and perspectives regarding the function of Polycomb complexes in shaping the stem cell genome architecture, and discuss how this function might be required to properly orchestrate transcriptional programs during differentiation.


Long-term outcomes in elderly survivors of in-hospital cardiac arrest.
Little is known about the long-term outcomes in elderly survivors of in-hospital cardiac arrest. We determined rates of long-term survival and readmission among survivors of in-hospital cardiac arrest and examined whether these outcomes differed according to demographic characteristics and neurologic status at discharge. We linked data from a national registry of inpatient cardiac arrests with Medicare files and identified 6972 adults, 65 years of age or older, who were discharged from the hospital after surviving an in-hospital cardiac arrest between 2000 and 2008. Predictors of 1-year survival and of readmission to the hospital were examined. One year after hospital discharge, 58.5% of the patients were alive, and 34.4% had not been readmitted to the hospital. The risk-adjusted rate of 1-year survival was lower among older patients than among younger patients (63.7%, 58.6%, and 49.7% among patients 65 to 74, 75 to 84, and ≥85 years of age, respectively; P<0.001), among men than among women (58.6% vs. 60.9%, P=0.03), and among black patients than among white patients (52.5% vs. 60.4%, P=0.001). The risk-adjusted rate of 1-year survival was 72.8% among patients with mild or no neurologic disability at discharge, as compared with 61.1% among patients with moderate neurologic disability, 42.2% among those with severe neurologic disability, and 10.2% among those in a coma or vegetative state (P<0.001 for all comparisons). Moreover, 1-year readmission rates were higher among patients who were black, those who were women, and those who had substantial neurologic disability (P<0.05 for all comparisons). These differences in survival and readmission rates persisted at 2 years. At 3 years, the rate of survival among survivors of in-hospital cardiac arrest was similar to that of patients who had been hospitalized with heart failure and were discharged alive (43.5% and 44.9%, respectively; risk ratio, 0.98; 95% confidence interval, 0.95 to 1.02; P=0.35). Among elderly survivors of in-hospital cardiac arrest, nearly 60% were alive at 1 year, and the rate of 3-year survival was similar to that among patients with heart failure. Survival and readmission rates differed according to the demographic characteristics of the patients and neurologic status at discharge. (Funded by the American Heart Association and the National Heart, Lung, and Blood Institute.).


General anesthetic neurotoxicity in the young: Mechanism and prevention.
General anesthesia (GA) is usually considered to safely induce a reversible unconscious state allowing surgery to be performed without pain. A growing number of studies, in particular pre-clinical studies, however, demonstrate that general anesthetics can cause neuronal death and even long-term neurological deficits. Herein, we report our literature review and meta-analysis data of the neurological outcomes after anesthesia in the young. We also review available mechanistic and epigenetic data of GA exposure related to cognitive impairment per se and the potential preventive strategies including natural herbal compounds to attenuate those side effects. In summary, anesthetic-induced neurotoxicity may be treatable and natural herbal compounds and other medications may have great potential for such use but warrants further study before clinical applications can be initiated.


POMICS: A Simulation Disease Model for Timing Fungicide Applications in Management of Powdery Mildew of Cucurbits.
A weather-based simulation model, called Powdery Mildew of Cucurbits Simulation (POMICS), was constructed to predict fungicide application scheduling to manage powdery mildew of cucurbits. The model was developed on the principle that conditions favorable for Podosphaera xanthii, a causal pathogen of this crop disease, generate a number of infection cycles in a single growing season. The model consists of two components that (i) simulate the disease progression of P. xanthii in secondary infection cycles under natural conditions and (ii) predict the disease severity with application of fungicides at any recurrent disease cycles. The underlying environmental factors associated with P. xanthii infection were quantified from laboratory and field studies, and also gathered from literature. The performance of the POMICS model when validated with two datasets of uncontrolled natural infection was good (the mean difference between simulated and observed disease severity on a scale of 0 to 5 was 0.02 and 0.05). In simulations, POMICS was able to predict high- and low-risk disease alerts. Furthermore, the predicted disease severity was responsive to the number of fungicide applications. Such responsiveness indicates that the model has the potential to be used as a tool to guide the scheduling of judicious fungicide applications.


Infection-stimulated infraosseus inflammation and bone destruction is increased in P-/E-selectin knockout mice.
Infections of the dental pulp commonly result in infraosseus inflammation and bone destruction. However, the role of phagocytic leucocytes in the pathogenesis of pulpal infections has been uncertain. In this work we used P/E-/- selectin-deficient mice, which lack rolling adhesion of leucocytes to endothelium and mimic the human syndrome, leucocyte adhesion deficiency II (LAD-II), to test the hypothesis that phagocytic leucocytes protect against pulpal infection and subsequent periapical infraosseus bone resorption. P/E-/- mice and P/E+/+ wild-type controls were subjected to surgical pulp exposure, and both groups were infected with a mixture of pulpal pathogens including Prevotella intermedia, Fusobacterium nucleatum, Peptostreptococcus micros and Streptococcus intermedius. Animals were killed after 20 days, and the extent of infraosseus bone destruction was quantified by histomorphometry. In two separate experiments, P/E-/- mice had significantly greater bone resorption than P/E+/+ controls. The increased bone destruction correlated with a twofold decrease in polymorphonuclear (PMN) infiltration into periapical inflammatory tissues of P/E-/- mice. P/E-/- mice had higher tissue levels of the bone resorptive cytokine, interleukin (IL)-1alpha. Tissue levels of IL-2, IL-4, IL-10, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) were all higher in P/E-/- mice, but the increases were not statistically significant. Only IL-12 was higher in P/E+/+ mice, possibly reflecting a greater number of infiltrating monocytes in wild-type mice. These findings demonstrate that phagocytic leucocytes are protective in this model, and suggest that elevated expression of inflammatory cytokines is responsible for the observed bone destruction.


Metabolism of the vacuolar pathogen Legionella and implications for virulence.
Legionella pneumophila is a ubiquitous environmental bacterium that thrives in fresh water habitats, either as planktonic form or as part of biofilms. The bacteria also grow intracellularly in free-living protozoa as well as in mammalian alveolar macrophages, thus triggering a potentially fatal pneumonia called "Legionnaires' disease." To establish its intracellular niche termed the "Legionella-containing vacuole" (LCV), L. pneumophila employs a type IV secretion system and translocates ~300 different "effector" proteins into host cells. The pathogen switches between two distinct forms to grow in its extra- or intracellular niches: transmissive bacteria are virulent for phagocytes, and replicative bacteria multiply within their hosts. The switch between these forms is regulated by different metabolic cues that signal conditions favorable for replication or transmission, respectively, causing a tight link between metabolism and virulence of the bacteria. Amino acids represent the prime carbon and energy source of extra- or intracellularly growing L. pneumophila. Yet, the genome sequences of several Legionella spp. as well as transcriptome and proteome data and metabolism studies indicate that the bacteria possess broad catabolic capacities and also utilize carbohydrates such as glucose. Accordingly, L. pneumophila mutant strains lacking catabolic genes show intracellular growth defects, and thus, intracellular metabolism and virulence of the pathogen are intimately connected. In this review we will summarize recent findings on the extra- and intracellular metabolism of L. pneumophila using genetic, biochemical and cellular microbial approaches. Recent progress in this field sheds light on the complex interplay between metabolism, differentiation and virulence of the pathogen.


The argonaut shell: gas-mediated buoyancy control in a pelagic octopus.
Argonauts (Cephalopoda: Argonautidae) are a group of rarely encountered open-ocean pelagic octopuses with benthic ancestry. Female argonauts inhabit a brittle 'paper nautilus' shell, the role of which has puzzled naturalists for millennia. The primary role attributed to the shell has been as a receptacle for egg deposition and brooding. Our observations of wild argonauts have revealed that the thin calcareous shell also functions as a hydrostatic structure, employed by the female argonaut to precisely control buoyancy at varying depths. Female argonauts use the shell to 'gulp' a measured volume of air at the sea surface, seal off the captured gas using flanged arms and forcefully dive to a depth where the compressed gas buoyancy counteracts body weight. This process allows the female argonaut to attain neutral buoyancy at depth and potentially adjust buoyancy to counter the increased (and significant) weight of eggs during reproductive periods. Evolution of this air-capture strategy enables this negatively buoyant octopus to survive free of the sea floor. This major shift in life mode from benthic to pelagic shows strong evolutionary parallels with the origins of all cephalopods, which attained gas-mediated buoyancy via the closed-chambered shells of the true nautiluses and their relatives.


Feasibility study evaluating a veterinary point-of-care urine culture system for diagnosing septic peritonitis.
To evaluate the use of a veterinary point-of-care urine culture system (POCUCS) for the diagnosis of septic peritonitis. Prospective feasibility study performed between August 2017 and April 2018. Private referral hospital. Twenty samples of naturally occurring canine peritoneal effusion collected via aseptic abdominocentesis. Point-of-care urine culture systems were inoculated and incubated according to manufacturer's instructions. The presence of bacterial growth, estimation of colony-forming units/mL of bacteria, and organism identification were recorded. Bacterial growth and organism identification on POCUCS were compared to an aerobic culture performed at a commercial microbiology laboratory. Serial dilution and subsequent culture on a POCUCS of a confirmed Escherichia coli infected peritoneal effusion and negative control sample were performed to determine the lowest concentration of bacteria detectable. There were 10 septic and 10 aseptic samples of peritoneal effusion confirmed by aerobic laboratory culture. Of the 10 culture-positive samples, 8 were culture-positive on the POCUCS. The sensitivity and specificity of the POCUCS for the detection of bacteria in peritoneal effusion were 80.0% and 100%, respectively. The POCUCS lowest limit of detectable bacteria in peritoneal effusion was 1000 CFUs/mL. The POCUCS evaluated in this study was less sensitive and less rapid for diagnosing septic peritonitis than blood glucose to peritoneal effusion glucose ratio and plasma lactate to peritoneal effusion glucose ratio. This POCUCS is not recommended as a tool for diagnosing septic peritonitis.


International Brainstorming Meeting on Etiologic and Risk Factors of Peri-implantitis, Montegrotto (Padua, Italy), August 2014.
The emerging literature has recently reported an alarming increase in peri-implantitis. This disease is typically described as the result of an imbalance between host response and bacterial load, supported by gram-negative anaerobic microflora. The current literature on the prevention and treatment of peri-implantitis does not allow for the extraction of applicable clinical information. In fact, the lack of efficacy of the current treatment methods may be a result of insufficient understanding of the biology. The aim of this position paper was to try to reevaluate the etiopathogenesis of peri-implantitis, highlighting the principal clinically induced triggering factors of the disease. The consensus conference provided strong evidence to suggest that a different microbiologic flora (slightly different from that collected around teeth affected by periodontitis) could support peri-implantitis. However, the evidence to support a consensus statement regarding clinically triggering factors (surgical, prosthetic, and biomechanical) for peri-implantitis is only of moderate strength (cohort studies or consistent results from long-term, well-populated case series). Expert opinion led the consensus group to support the following: rectifying the number of peri-implant inflammatory situations caused by surgical, restorative, or material complications may lower the number of infections to a more realistic figure and may suggest different and more appropriate treatment plans. At the same time, it can be stated that implant material, shape and surface characteristics, procedures and biomaterials used for bone augmentation, and incorrect prosthetic procedures and biomechanical plans could also be risk factors for the occurrence and progression of periimplantitis.


Progressive kyphoscoliosis associated with tethered cord treated by posterior vertebral column resection: a case report.
STUDY DESIGN.: A case report. OBJECTIVES.: To report a case of progressive kyphoscoliosis associated with a tethered cord that was corrected by posterior vertebral column resection after complicated untethering surgery. SUMMARY OF BACKGROUND DATA.: There have been few clinical reports on posterior vertebral column resection conducted for severe deformity associated with a tethered cord. METHODS.: A patient with progressive kyphoscoliosis associated with a tethered cord first underwent untethering surgery, resulting in neurologic deterioration. Posterior vertebral column resection was performed to correct the kyphoscoliosis while shortening the spinal column to prevent the spinal cord from stretch injury. RESULTS.: Good correction of kyphoscoliosis was obtained without further neurologic deterioration. The Cobb angles of scoliosis was 103 degrees before surgery and 25 degrees after surgery (correction rate; 75.7%), and that of kyphosis was 90 degrees and 36 degrees , respectively (correction rate; 60.0%). CONCLUSION.: Correction of progressive kyphoscoliosis associated with a tethered cord can be achieved successfully by posterior vertebral column resection even after complicated untethering surgery.


The cryptic African wolf: Canis aureus lupaster is not a golden jackal and is not endemic to Egypt.
The Egyptian jackal (Canis aureus lupaster) has hitherto been considered a large, rare subspecies of the golden jackal (C. aureus). It has maintained its taxonomical status to date, despite studies demonstrating morphological similarities to the grey wolf (C. lupus). We have analyzed 2055 bp of mitochondrial DNA from C. a. lupaster and investigated the similarity to C. aureus and C. lupus. Through phylogenetic comparison with all wild wolf-like canids (based on 726 bp of the Cytochrome b gene) we conclusively (100% bootstrap support) place the Egyptian jackal within the grey wolf species complex, together with the Holarctic wolf, the Indian wolf and the Himalayan wolf. Like the two latter taxa, C. a. lupaster seems to represent an ancient wolf lineage which most likely colonized Africa prior to the northern hemisphere radiation. We thus refer to C. a. lupaster as the African wolf. Furthermore, we have detected C. a. lupaster individuals at two localities in the Ethiopian highlands, extending the distribution by at least 2,500 km southeast. The only grey wolf species to inhabit the African continent is a cryptic species for which the conservation status urgently needs assessment.


Management of human papillomavirus-related head and neck cancer.
Human papillomavirus (HPV)-related head and neck malignancies (HNMs) have become a serious health risk over the past 20 years. Despite decreases in non-HPV-related HNMs, the incidence of HPV-related HNMs has skyrocketed, and a new form of tumorigenesis is developing. HPV type 16 is the primary offender, and the majority of these tumors present in the oropharynx, with a smaller proportion in the larynx and oral cavity. While traditionally treated with surgery, the paradigm has shifted to more of a nonoperative chemoradiation therapy approach, with the hope of improving vital functions after therapy. Unfortunately, we continue to see significant dysphagia in these patients after treatment, and work is being done to improve outcomes. With the advent of transoral robotic surgery, we have again been able to reconsider treatment options for these patients, although it has been met with some skepticism and resistance. Here we discuss the scope of HPV-related HNMs, the treatment options and prognosis for the disease, and finally touch upon psychosocial issues related to HPV-related HNMs.


Ultrasound-triggered, flight-gated evasive maneuvers in the praying mantis Parasphendale agrionina. I. Free flight.
Free-flying male praying mantises Parasphendale agrionina (Gerst.) perform evasive maneuvers when stimulated by ultrasound and when attacked by hunting, echolocating bats. They do not, however, respond in any way when standing on a substratum. The maneuvers are graded in intensity with distance from the sound source: far from the source they are simple turns, whereas close to the source they are steep diving turns or spirals. The maneuvers are made under power, and the male's velocity doubles to almost 4 m s-1 by the end of a steep dive. The mantis does not show any directional preference. The behavioral threshold of 64 dB SPL and minimum latency to course change of 125 ms indicate that these mantises should have adequate time to evade bats using calls of greater than 85-90 dB SPL (at 10 cm). In field experiments with wild, hunting bats, P. agrionina successfully evaded capture in all five attacks to which they responded with evasive maneuvers. Out of three attacks on P. aeruginosa and three on a normally non-responding mantis, Miomantis paykullii Stål, in which there were no evasive maneuvers, the mantis was captured in five cases.


Bio-Ecological Features Update on Eleven Rare Cartilaginous Fish in the Central-Western Mediterranean Sea as a Contribution for Their Conservation.
Cartilaginous fish are commonly recognized as key species in marine ecosystems for their fundamental ecological role as top predators. Nevertheless, effective management plans for cartilaginous fish are still missing, due to the lack of knowledge on their abundance, distribution or even life-history. In this regard, this paper aims at providing new information on the life-history traits, such as age, maturity, reproductive period, in addition to diet characteristics of eleven rare cartilaginous fish inhabiting the Central-Western Mediterranean Sea belonging to the orders Chimaeriformes (Chimaera monstrosa), Hexanchiformes (Heptranchias perlo and Hexanchus griseus), Myliobatiformes (Aetomylaeus bovinus and Myliobatis aquila), Rajiformes (Dipturus nidarosiensis and Leucoraja circularis), Squaliformes (Centrophorus uyato, Dalatias licha and Oxynotus centrina) and Torpediniformes (Tetronarce nobiliana), useful for their assessment and for future management actions. Particularly, the present paper provides for the first time the age estimation of D. nidarosienis and L. circularis which were both found capable of becoming older than ten years. In addition, the present study updates the sizes of first maturity of C. uyato and D. licha, which appeared to be capable of reproducing earlier than what was previously hypothesized, representing very valuable information for a better understanding of these rare species populations status and, eventually, their conservation. On the basis of the stomach content analysis, it was possible to identify five different predator groups.


NAFLD fibrosis score: a prognostic predictor for mortality and liver complications among NAFLD patients.
To study whether the severity of liver fibrosis estimated by the nonalcoholic fatty liver disease (NAFLD) fibrosis score can predict all-cause mortality, cardiac complications, and/or liver complications of patients with NAFLD over long-term follow-up. A cohort of well-characterized patients with NAFLD diagnosed during the period of 1980-2000 was identified through the Rochester Epidemiology Project. The NAFLD fibrosis score (NFS) was used to separate NAFLD patients with and without advanced liver fibrosis. We used the NFS score to classify the probability of fibrosis as < -1.5 for low probability, > -1.5 to < 0.67 for intermediate probability, and > 0.67 for high probability. Primary endpoints included all-cause death and cardiovascular- and/or liver-related mortality. From the 479 patients with NAFLD assessed, 302 patients (63%) greater than 18 years old were included. All patients were followed, and medical charts were reviewed until August 31, 2009 or the date when the first primary endpoint occurred. By using a standardized case record form, we recorded a detailed history and physical examination and the use of statins and metformin during the follow-up period. A total of 302/479 (63%) NAFLD patients (mean age: 47 ± 13 year) were included with a follow-up period of 12.0 ± 3.9 year. A low probability of advanced fibrosis (NFS < -1.5 at baseline) was found in 181 patients (60%), while an intermediate or high probability of advanced fibrosis (NSF > -1.5) was found in 121 patients (40%). At the end of the follow-up period, 55 patients (18%) developed primary endpoints. A total of 39 patients (13%) died during the follow-up. The leading causes of death were non-hepatic malignancy (n = 13/39; 33.3%), coronary heart disease (CHD) (n = 8/39; 20.5%), and liver-related mortality (n = 5/39; 12.8%). Thirty patients had new-onset CHD, whereas 8 of 30 patients (27%) died from CHD-related causes during the follow-up. In a multivariate analysis, a higher NFS at baseline and the presence of new-onset CHD were significantly predictive of death (OR = 2.6 and 9.2, respectively; P < 0.0001). Our study showed a significant, graded relationship between the NFS, as classified into 3 subgroups (low, intermediate and high probability of liver fibrosis), and the occurrence of primary endpoints. The use of metformin or simvastatin for at least 3 mo during the follow-up was associated with fewer deaths in patients with NAFLD (OR = 0.2 and 0.03, respectively; P < 0.05). Additionally, the rate of annual NFS change in patients with an intermediate or high probability of advanced liver fibrosis was significantly lower than those patients with a low probability of advanced liver fibrosis (0.06 vs 0.09, P = 0.004). The annual NFS change in patients who died was significantly higher than those in patients who survived (0.14 vs 0.07, P = 0.03). At the end of the follow-up, we classified the patients into 3 subgroups according to the progression pattern of liver fibrosis by comparing the NFS at baseline to the NFS at the end of the follow-up period. Most patients were in the stable-fibrosis (60%) and progressive-fibrosis (37%) groups, whereas only 3% were in the regressive fibrosis. A higher NAFLD fibrosis score at baseline and a new onset of CHD were significantly predictive of death in patients with NAFLD.


A convergent functional architecture of the insula emerges across imaging modalities.
Empirical evidence increasingly supports the hypothesis that patterns of intrinsic functional connectivity (iFC) are sculpted by a history of evoked coactivation within distinct neuronal networks. This, together with evidence of strong correspondence among the networks defined by iFC and those delineated using a variety of other neuroimaging techniques, suggests a fundamental brain architecture detectable across multiple functional and structural imaging modalities. Here, we leverage this insight to examine the functional organization of the human insula. We parcellated the insula on the basis of three distinct neuroimaging modalities - task-evoked coactivation, intrinsic (i.e., task-independent) functional connectivity, and gray matter structural covariance. Clustering of these three different covariance-based measures revealed a convergent elemental organization of the insula that likely reflects a fundamental brain architecture governing both brain structure and function at multiple spatial scales. While not constrained to be hierarchical, our parcellation revealed a pseudo-hierarchical, multiscale organization that was consistent with previous clustering and meta-analytic studies of the insula. Finally, meta-analytic examination of the cognitive and behavioral domains associated with each of the insular clusters obtained elucidated the broad functional dissociations likely underlying the topography observed. To facilitate future investigations of insula function across healthy and pathological states, the insular parcels have been made freely available for download via http://fcon_1000.projects.nitrc.org, along with the analytic scripts used to perform the parcellations.


Induction of cytochrome P-450 isozyme 3a (P-450IIE1) in rabbit olfactory mucosa by ethanol and acetone.
Cytochrome P-450 isozyme 3a, the alcohol-inducible form of cytochrome P-450 (P-450IIE1), was previously identified in rabbit nasal microsomes with the use of immunochemical techniques; the occurrence of this cytochrome in the nasal mucosa was subsequently confirmed through RNA hybridization experiments. However, in contrast to the well established inducibility of isozyme 3a in liver and kidney by alcohol treatment of the animals, no induction was observed in the nasal tissue with the use of a polyclonal anti-3a antibody for immunochemical quantitation. Recently, two new P-450 isozymes, designated NMa and NMb, were identified in rabbit nasal microsomes, and were found to have overlapping substrate specificity with isozyme 3a. Moreover, the two new cytochromes cross-react with the polyclonal anti-3a antibody that was used in the earlier study for quantitation of nasal isozyme 3a. These recent findings invalidate our previous conclusion that isozyme 3a is not induced by ethanol treatment of rabbits. In the present study, immunoblot quantitation of isozyme 3a was performed with a monoclonal anti-3a antibody that does not recognize either NMa or NMb, and the nasal microsomal metabolism of butanol was examined at various substrate concentrations. We have found that the level of isozyme 3a protein in nasal mucosa is elevated about 2-fold after treatment of the animals with ethanol and about 6-fold after treatment with acetone. Furthermore, corresponding increases in the rate of microsomal butanol oxidation were observed at low substrate concentrations. Thus, we conclude that P-450 isozyme 3a is, in fact, inducible in the nasal tissues by ethanol or acetone treatment of rabbits.


A dual acting compound releasing nitric oxide (NO) and ibuprofen, NCX 320, shows significant therapeutic effects in a mouse model of muscular dystrophy.
A resolutive therapy for muscular dystrophies, a heterogeneous group of genetic diseases leading to muscular degeneration and in the severe forms to death, is still lacking. Since inflammation and defects in nitric oxide generation are recognized key pathogenic events in muscular dystrophy, we have analysed the effects of a derivative of ibuprofen, NCX 320, belonging to the class of cyclooxygenase inhibiting nitric oxide donator (CINOD), in the α-sarcoglycan null mice, a severe mouse model of dystrophy. NCX 320 was administered daily in the diet for 8months starting 1month from weaning. Muscle functional recovery was evaluated by free wheel and treadmill tests at 8months. Serum creatine kinase activity, as well as the number of diaphragm inflammatory infiltrates and necrotic fibres, was measured as indexes of skeletal muscle damage. Muscle regeneration was evaluated in diaphragm and tibialis anterior muscles, measuring the numbers of centronucleated fibres and of myogenic precursor cells. NCX 320 mitigated muscle damage, reducing significantly serum creatine kinase activity, the number of necrotic fibres and inflammatory infiltrates. Moreover, NCX 320 stimulated muscle regeneration increasing significantly the number of myogenic precursor cells and regenerating fibres. All these effects concurred in inducing a significant improvement of muscle function, as assessed by both free wheel and treadmill tests. These results describe the properties of a new compound incorporating nitric oxide donation together with anti-inflammatory properties, showing that it is effective in slowing muscle dystrophy progression long term. Of importance, this new compound deserves specific attention for its potential in the therapy of muscular dystrophy given that ibuprofen is well tolerated in paediatric patients and with a profile of safety that makes it suitable for chronic treatment such as the one required in muscular dystrophies.


Androgen-targeted therapy in men with prostate cancer: evolving practice and future considerations.
Androgen deprivation therapy (ADT) is foundational in the management of advanced prostate cancer (PCa) and has benefitted from a recent explosion in scientific advances. These include approval of new therapies that suppress testosterone (T) levels or inactivate its function, improvements in diagnostic and assay technologies, identification of lower therapeutic targets for T, discovery of the relevance of germline genetic mutations and identification of the benefits of sequential and combination therapies. This review discusses the clinical profiles of the most up-to-date options for ADT, best practices for managing patients with advanced PCa and future directions in therapy. Modern assay technologies reveal that bilateral orchiectomy results in a serum T level of approximately 15 ng/dL as compared to the historical definition of castration of T < 50 ng/dL. Evidence shows that lowering T levels to <20 ng/dL improves patient survival and delays disease progression. Routine monitoring of T in addition to prostate-specific antigen throughout treatment is important to ensure continuing efficacy of T suppression. New drugs that inhibit androgen signaling in combination with traditional ADT suppress T activity to near zero and have significantly improved patient survival. When personalizing ADT regimens physicians should consider a number of factors including initiation and duration of ADT, monitoring of T levels and PSA, the possibility of switching monotherapies if a patient does not achieve adequate T suppression, and consideration of intermittent vs. continuous ADT according to patients' lifestyles, comorbidities, risk factors and tolerance to treatment.


Combining lapatinib and pertuzumab to overcome lapatinib resistance due to NRG1-mediated signalling in HER2-amplified breast cancer.
Acquired resistance to lapatinib, an inhibitor of EGFR and HER2 kinases, is common. We found that reactivation of EGFR, HER2 and HER3 occurred within 24 hours of lapatinib treatment after their initial dephosphorylation. This was associated with increased expression of NRG1 in cells treated with lapatinib. Exogenous NRG1 partially rescued breast cancer cells from growth inhibition by lapatinib. In addition, both parental and lapatinib-resistant breast cancer cells were sensitive to SGP1, which inhibits binding of NRG1 and other HER3 ligands. Addition of pertuzumab to lapatinib further inhibited NRG1-induced signalling, which was not fully inhibited by either drug alone. In animal model, a combination of pertuzumab to lapatinib induced a greater tumor regression than either lapatinib or pertuzumab monotherapy. This novel combination treatment may provide a promising strategy in clinical HER2-targeted therapy and may inhibit a subset of lapatinib-resistant breast cancer, although the group of patients that will respond to this therapy requires further stratification.


Implementation of a genomic medicine multi-disciplinary team approach for rare disease in the clinical setting: a prospective exome sequencing case series.
A multi-disciplinary approach to promote engagement, inform decision-making and support clinicians and patients is increasingly advocated to realise the potential of genome-scale sequencing in the clinic for patient benefit. Here we describe the results of establishing a genomic medicine multi-disciplinary team (GM-MDT) for case selection, processing, interpretation and return of results. We report a consecutive case series of 132 patients (involving 10 medical specialties with 43.2% cases having a neurological disorder) undergoing exome sequencing over a 10-month period following the establishment of the GM-MDT in a UK NHS tertiary referral hospital. The costs of running the MDT are also reported. In total 76 cases underwent exome sequencing following triage by the GM-MDT with a clinically reportable molecular diagnosis in 24 (31.6%). GM-MDT composition, operation and rationale for whether to proceed to sequencing are described, together with the health economics (cost per case for the GM-MDT was £399.61), the utility and informativeness of exome sequencing for molecular diagnosis in a range of traits, the impact of choice of sequencing strategy on molecular diagnostic rates and challenge of defining pathogenic variants. In 5 cases (6.6%), an alternative clinical diagnosis was indicated by sequencing results. Examples were also found where findings from initial genetic testing were reconsidered in the light of exome sequencing including TP63 and PRKAG2 (detection of a partial exon deletion and a mosaic missense pathogenic variant respectively); together with tissue-specific mosaicism involving a cytogenetic abnormality following a normal prenatal array comparative genomic hybridization. This consecutive case series describes the results and experience of a multidisciplinary team format that was found to promote engagement across specialties and facilitate return of results to the responsible clinicians.


Molecular physiology and breeding at the crossroads of cold hardiness improvement.
Alfalfa (Medicago sativa L.) is a major forage legume grown extensively worldwide with important agronomic and environmental attributes. Insufficient cold hardiness is a major impediment to its reliable production in northern climates. Improvement of freezing tolerance using conventional breeding approaches is slowed by the quantitative nature of inheritance and strong interactions with the environment. The development of gene-based markers would facilitate the identification of genotypes with superior stress tolerance. Successive cycles of recurrent selection were applied using an indoor screening method to develop populations with significantly higher tolerance to freezing (TF). Bulk segregant analysis of heterogeneous TF populations identified DNA variations that are progressively enriched in frequency in response to selection. Polymorphisms resulting from intragenic variations within a dehydrin gene were identified and could potentially lead to the development of robust selection tools. Our results illustrate the benefits of feedback interactions between germplasm development programs and molecular physiology for a deeper understanding of the molecular and genetic bases of cold hardiness.


Comparison of Conventional, Revascularized, and Bioengineered Methods of Recurrent Laryngeal Nerve Reconstruction.
Damage to the recurrent laryngeal nerve (RLN) is highly detrimental to voice, swallow, and cough. The optimal method for reconstitution of a nerve gap after injury is unknown. To evaluate multiple methods of RLN reconstruction. This study used an established canine model of RLN injury to examine purpose-bred, conditioned, female, 20-kg mongrel hounds at Washington University. A total of 32 dogs were examined, with 63 experiments performed. Surgical transection or excision of the RLN with reconstruction by multiple methods. Six months after injury repair, laryngeal adductor pressures (LAPs), spontaneous and stimulable movement, and graft axon counts by histologic analysis were assessed. Simple RLN transection with direct neurorrhaphy provided a mean (SD) recovery of 55.5% (12.5%) of baseline LAPs (P = .18 for comparison of LAP recovery among cases from the conventional nerve graft [39.4% (22.2%)]; P = .63 for comparison of LAP recovery among cases from the reverse autograft [60.8% (27.5%)]). Revascularized grafts provided a recovery of 54.5% (46.4%) while short and long acellular grafts provided recoveries of 60.4% (NA) and 39.5% (17.0%). Two of 11 polyglycolic acid reconstructions provided a measurable LAP with a mean (SD) recovery of 37.1% (8.9%) of baseline. Reconstruction with a neural conduit in any condition provided no measurable LAP recovery. Conventional nerve grafting resulted in no significant difference in recovery of LAP function compared with simple neurorrhaphy or reverse autograft. Conventional and revascularized nerve grafts provided similar recovery. The use of bioengineered acellular nerve grafts or nerve conduits for reconstruction resulted in poor recovery of function.


Combined treatment of adenosine nucleoside inhibitor NITD008 and histone deacetylase inhibitor vorinostat represents an immunotherapy strategy to ameliorate West Nile virus infection.
West Nile virus (WNV), a member of the Flaviviridae family, is the leading cause of viral encephalitis in the United States. Despite efforts to control the spread of WNV, there has been an increase in the number of outbreaks and clinical cases with neurological problems. There are no antiviral compounds currently in trials for WNV. NITD008 is an adenosine analogue inhibitor that interrupts the RNA-dependent RNA polymerase of flaviviruses. Previous studies demonstrated NITD008 as a potent antiviral for dengue virus, however this drug was associated with preclinical toxicity. The ability of NITD008 to block WNV replication is only shown in Vero cells. Neuroinflammation is also a major cause of the WNV-associated pathology, therefore we evaluated the effect of NITD008 and a newly characterized anti-inflammatory drug vorinostat (SAHA), a histone deacetylase inhibitor, on WNV replication and disease progression in a mouse model. When administered at 10 and 25mg/kg at days 1-6 after WNV infection in C57BL/6 mice, NITD008 conferred complete protection from clinical symptoms and death, which correlated with reduced viral load in the serum and restriction of virus-CNS entry. Delay of NITD008 treatment to days 3-6 and days 5-9 after infection, when WNV replication was high in the periphery and brain, resulted in the gradual loss of protection against WNV infection. However, co-treatment with SAHA and NITD008 during the CNS phase of disease improved disease outcome significantly by reducing inflammation and neuronal death. Our results support potential synergistic effect of combination therapy of NITD008 with SAHA for the treatment of WNV encephalitis.


A novel mechanism of inhibition of high-voltage activated calcium channels by α-conotoxins contributes to relief of nerve injury-induced neuropathic pain.
α-Conotoxins that are thought to act as antagonists of nicotinic acetylcholine receptors (nAChRs) containing α3-subunits are efficacious in several preclinical models of chronic pain. Potent interactions of Vc1.1 with other targets have suggested that the pain-relieving actions of α-conotoxins might be mediated by either α9α10 nAChRs or a novel GABA(B) receptor-mediated inhibition of N-type calcium channels. Here we establish that three α-conotoxins, Vc1.1, AuIB and MII have distinct selectivity profiles for these three potential targets. Their potencies after intramuscular administration were then determined for reversal of allodynia produced by partial nerve ligation in rats. Vc1.1, which potently inhibits α9α10 nAChRs and GABA(B)/Ca(2+) channels but weakly blocks α3β2 and α3β4 nAChRs, produced potent, long-lasting reversal of allodynia that were prevented by pre-treatment with the GABA(B) receptor antagonist, SCH50911. α-Conotoxin AuIB, a weak α3β4 nAChR antagonist, inhibited GABA(B)/Ca(2+) channels but did not act on α9α10 nAChRs. AuIB also produced reversal of allodynia. These findings suggest that GABA(B) receptor-dependent inhibition of N-type Ca(2+) channels can mediate the sustained anti-allodynic actions of some α-conotoxins. However, MII, a potent α3β2 nAChR antagonist but inactive on α9α10 and α3β4 nAChRs and GABA(B)/Ca(2+) channels, was demonstrated to have short-acting anti-allodynic action. This suggests that α3β2 nAChRs may also contribute to reversal of allodynia. Together, these findings suggest that inhibition of α9α10 nAChR is neither necessary nor sufficient for relief of allodynia and establish that α-conotoxins selective for GABA(B) receptor-dependent inhibition of N-type Ca(2+) channels relieve allodynia, and could therefore be developed to manage chronic pain.


Can performance-based incentives improve motivation of nurses and midwives in primary facilities in northern Ghana? A quasi-experimental study.
Lack of an adequate and well-performing health workforce has emerged as the biggest barrier to scaling up health services provision in sub-Saharan Africa. As the global community commits to the Sustainable Development Goals and universal health coverage, health workforce challenges are critical. In northern Ghana, performance-based incentives (PBIs) were introduced to improve health worker motivation and service quality. The goal of this study was to determine the impact of PBIs on maternal health worker motivation in two districts in northern Ghana. A quasi-experimental study design with pre- and post-intervention measurement was used. PBIs were implemented for 2 years in six health facilities in Kassena-Nankana District with six health facilities in Builsa District serving as comparison sites. Fifty pre- and post-intervention structured interviews and 66 post-intervention in-depth interviews were conducted with health workers. Motivation was assessed using constructs for job satisfaction, pride, intrinsic motivation, timelines/attendance, and organisational commitment. Quantitative data were analysed to determine changes in motivation between intervention and comparison facilities pre- and post-intervention using STATA™ version 13. Qualitative data were analysed thematically using NVivo 10 to explore possible reasons for quantitative findings. PBIs were associated with slightly improved maternal health worker motivation. Mean values for overall motivation between intervention and comparison health workers were 0.6 versus 0.7 at baseline and 0.8 versus 0.7 at end line, respectively. Differences at baseline and end line were 0.1 (p=0.40 and p=0.50 respectively), with an overall 0.01 difference in difference (p=0.90). Qualitative interviews indicated that PBIs encouraged health workers to work harder and be more punctual, increasing reported pride and job satisfaction. The results contribute evidence on the effects of PBIs on motivational constructs among maternal health workers in primary care facilities in northern Ghana. PBIs appeared to improve motivation, but not dramatically, and the long-term and unintended effects of their introduction require additional study.


The construct validity of the work-related flow inventory in a sample of Australian workers.
The purpose of this study was to examine the construct validity of the WOrk-reLated Flow inventory (WOLF; Bakker, 2008). This instrument was administered to 711 men and women who were working in Queensland, Australia. The results from the confirmatory factor analysis showed that the WOLF has moderately acceptable construct validity, with the three-factor model being a borderline fit to the data. Tests of the convergent validity of the WOLF yielded satisfactory results. However, the analysis of the discriminant validity of the WOLF showed that the instrument poorly discriminated between work enjoyment and intrinsic work motivation. Follow-up exploratory factor analysis, using recommended procedures for determining the number of factors to extract, revealed a two-factor solution, with the work enjoyment and intrinsic work motivation items loading on the same factor. Drawing on literature on psychological flow and motivation, as well as the findings of the present study, questions are raised over the adequacy of the conceptual basis of the three-factor model of work-related flow, the discriminant validity of the WOLF subscales, and the appropriateness of the wording of several of this measure's items. Using alternative methods and measures to investigate flow in work settings is recommended.


Anticoagulation management in remote primary care.
To examine anticoagulation management at the Bella Coola Medical Clinic in British Columbia. Charts of all patients in the Bella Coola Valley receiving warfarin were assessed. Data were analyzed using Microsoft Excel. Bella Coola Medical Clinic on the remote central coast of British Columbia. Twenty-one patients at the Bella Coola Medical Clinic who were receiving warfarin. All international normalized ratio (INR) tests over the preceding 12 months were examined for results, time elapsed since previous test, and interval until next scheduled test. An in-range INR rate of 60% is considered acceptable for anticoagulation services. The clinic had performed 406 INR tests on these 21 patients over the last 12 months. We found that 53% of all INR results fell strictly within the recommended therapeutic range. The relative success of anticoagulation management in Bella Coola probably results from several factors. For instance, physicians usually responded to out-of-range INR results with close monitoring: in 71% of cases, follow-up tests were scheduled within 1 week. On average, patients attended 77% of these visits on schedule; 58% of all out-of-range INR results were followed up with retesting within 1 week. Our results suggest that primary care physicians can manage anticoagulation adequately, even in remote settings.


Constructing reduction-sensitive PEGylated NIRF mesoporous silica nanoparticles via a one-pot Passerini reaction for photothermal/chemo-therapy.
In this work, we decorated the surface of MSNs by a one-pot Passerini reaction using near-infrared fluorescence BODIPY and poly(ethylene glycol) with an aldehyde group (PEG-CHO) for the first time. Doxorubicin (Dox) was loaded to demonstrate the reduction response. The formed MSNs neatly integrate chemotherapy, photothermal therapy and near-infrared fluorescence (NIRF) imaging.


Shift workers have similar leisure-time physical activity levels as day workers but are more sedentary at work.
Objective Physical inactivity has been hypothesized as an underlying factor for the association between shift work and adverse health outcomes. We compared leisure-time and occupational physical activity and sedentary behavior between day, night, and non-night shift workers. Methods We identified 612 day workers, 139 night shift workers and 61 non-night shift workers aged 18-65 years (54% men) in two Danish studies: the New method for Objective Measurements of physical Activity in Daily living (NOMAD) and the Danish Physical ACTivity cohort with Objective measurements (DPhacto) between 2011-2013. Sedentary behavior, light, and moderate-to-vigorous physical activity were measured using an accelerometer. Physical activity was expressed as percentage of leisure and work time spent in each activity. Linear regression analyses were used to test differences in physical activity and sedentary behavior between day, night, and non-night shift workers. Results No differences in leisure-time sedentary behavior and physical activity were observed between day and shift workers (P>0.05). Non-night shift workers spent 7.2% [95% confidence interval (95% CI) 2.3-12.1) more time in occupational sedentary behavior than day workers and 5.9% (95% CI -10.1- -1.7) and 1.9% (95% CI -3.7- -0.2) less time in occupational light and moderate-to-vigorous physical activity, respectively. Compared to day workers, night shift workers spent 4.3% (95% CI 2.4-6.1) more time at work in uninterrupted sedentary periods of ≥30 minutes. Conclusions Shift workers had similar leisure-time physical activity patterns as day workers, but were more sedentary at work. Future research should elucidate whether occupational physical inactivity and sedentary behavior contributes to shift work-related adverse health effects.


Role of heme oxygenase-1 and Kupffer cells in the production of bilirubin in the rat liver.
Heme oxygenase (HO)-1, the heme-degrading enzyme in macrophages, plays a key role in bilirubin metabolism. HO-1 is expressed in various tissue macrophages, especially Kupffer cells. This study aimed to examine the roles of macrophages and HO-1 in the modulation of heme catabolism in rat livers. Rats treated with or without liposome-encapsulated dichloromethylene diphosphonate, a macrophage-depleting reagent, were administered with heat-denatured red blood cells (h-RBC), and the time course of the biliary output of bilirubin and the expressions of HO-1 mRNA and protein were monitored. Immunohistochemistry in the control rat liver revealed that Kupffer cells constitute a major cellular component expressing HO-1, while hepatocytes exhibited little expression. The levels of HO-1 expression in Kupffer cells were elevated immediately after injection of h-RBC. In Kupffer cell-depleted livers, however, HO-1-expressing cells were not detected even after h-RBC administration. HO-1 mRNA levels were elevated at 2 h after administration of h-RBC in control rat livers, while they were very low in Kupffer cell-depleted rat livers. The control and Kupffer cell-depleted groups exhibited distinct time courses of biliary bilirubin excretion. In the untreated control rats, total bilirubin excretion increased about two-fold at 5 h after h-RBC administration. In contrast, the Kupffer cell-depleting treatment decreased the level of bilirubin production; administration of h-RBC to Kupffer cell-depleted rats did not accelerate the generation of bilirubin. These results suggest that Kupffer cells serve both as a sensor for scenesent RBC clearance and an effector that upregulates heme-degrading capacity and bilirubin production.


Average volume of alcohol consumption, patterns of drinking, and all-cause mortality: results from the US National Alcohol Survey.
The objective of this study was to investigate the effects of an average volume of alcohol consumption and drinking patterns on all-cause mortality. The sample (n = 5,072) was drawn from the 1984 National Alcohol Survey, representative of the US population living in households. Follow-up time was until the end of 1995, with 532 people deceased during this period. The authors found a significant influence of drinking alcohol on mortality with a J-shaped association for males and an insignificant relation of the same shape for females. When the largest categories of equivalent average volume of consumption were divided into people with and without heavy drinking occasions, serving as an indicator of drinking pattern, this differentiation proved important in predicting mortality. Light to moderate drinkers had higher mortality risks when they reported heavy drinking occasions (defined by either eight drinks per occasion or getting drunk at least monthly). Similarly, when the category of exdrinkers was divided into people who did or did not report heavy drinking occasions in the past, people with heavy drinking occasions had a higher mortality risk. Finally, indicating alcohol problems in the past was related to higher mortality risk. Results emphasized the importance of routinely including measures of drinking patterns into future epidemiologic studies on alcohol-related mortality.


Risk predictive modelling for diabetes and cardiovascular disease.
Absolute risk models or clinical prediction models have been incorporated in guidelines, and are increasingly advocated as tools to assist risk stratification and guide prevention and treatments decisions relating to common health conditions such as cardiovascular disease (CVD) and diabetes mellitus. We have reviewed the historical development and principles of prediction research, including their statistical underpinning, as well as implications for routine practice, with a focus on predictive modelling for CVD and diabetes. Predictive modelling for CVD risk, which has developed over the last five decades, has been largely influenced by the Framingham Heart Study investigators, while it is only ∼20 years ago that similar efforts were started in the field of diabetes. Identification of predictive factors is an important preliminary step which provides the knowledge base on potential predictors to be tested for inclusion during the statistical derivation of the final model. The derived models must then be tested both on the development sample (internal validation) and on other populations in different settings (external validation). Updating procedures (e.g. recalibration) should be used to improve the performance of models that fail the tests of external validation. Ultimately, the effect of introducing validated models in routine practice on the process and outcomes of care as well as its cost-effectiveness should be tested in impact studies before wide dissemination of models beyond the research context. Several predictions models have been developed for CVD or diabetes, but very few have been externally validated or tested in impact studies, and their comparative performance has yet to be fully assessed. A shift of focus from developing new CVD or diabetes prediction models to validating the existing ones will improve their adoption in routine practice.


Serologic Reactivity Reflects Clinical Expression of Ulcerative Colitis in Children.
In contrast to pediatric Crohn's disease (CD), little is known in pediatric ulcerative colitis (UC) about the relationship between disease phenotype and serologic reactivity to microbial and other antigens. The aim of this study was to examine disease phenotype and serology in a well-characterized inception cohort of children newly diagnosed with UC during the PROTECT Study (Predicting Response to Standardized Pediatric Colitis Therapy). Patients were recruited from 29 participating centers. Demographic, clinical, laboratory, and serologic (pANCA, ASCA IgA/IgG, Anti-CBir1, and Anti-OmpC) data were obtained from children 4-17 years old with UC. Sixty-five percent of the patients had positive serology for pANCA, with 62% less than 12 years old and 66% 12 years old or older. Perinuclear anti-neutrophil cytoplasmic antibodies did not correspond to a specific phenotype though pANCA ≥100, found in 19%, was strongly associated with pancolitis (P = 0.003). Anti-CBir1 was positive in 19% and more common in younger children with 32% less than 12 years old as compared with 14% 12 years old or older (P < 0.001). No association was found in any age group between pANCA and Anti-CBir1. Relative rectal sparing was more common in +CBir1, 16% versus 7% (P = 0.02). Calprotectin was lower in Anti-CBir1+ (Median [IQR] 1495 mcg/g [973-3333] vs 2648 mcg/g [1343-4038]; P = 0.04). Vitamin D 25-OH sufficiency was associated with Anti-CBir1+ (P = 0.0009). The frequency of pANCA in children was consistent with adult observations. High titer pANCA was associated with more extensive disease, supporting the idea that the magnitude of immune reactivity may reflect disease severity. Anti-CBir1+ was more common in younger ages, suggesting host-microbial interactions may differ by patient age.


Being in a standstill-of-life: women's experience of being diagnosed with systemic lupus erythematosus: a hermeneutic-phenomenological study.
Systemic lupus erythematosus (SLE) is a highly unpredictable and potentially lethal disease which ultimately challenges identity, future and the meaning of life. In a caring context, the experience of good health is perceived to be a balance between biomedical health and the existential experience of having a good life. This balance is jeopardised in the face of severe chronic illness and leads to extensive suffering if not handled carefully. Research suggests that patients suffering from severe chronic illness need support on an existential level, but also emphasises that, given its elusive nature, caring for the existential dimension is difficult to manage. This paper explores the experience of being diagnosed with SLE as an existential phenomenon. Through repeated phenomenological and hermeneutic interviews with 15 women conducted from 2013 to 2015, data concerning the diagnostic phase of SLE were analysed using Van Manen's phenomenology of practice. The essence was found to be a standstill in life comprehended through three inter-related themes: standing in a swirl of events, standing on uneven ground and standing at a turning point with oneself and others. The paper elucidates how existential life phenomena are lived, during the course of being diagnosed. In conclusion, it provides an ethical awareness of how a standstill in life is lived and of the patients' existential transition during the diagnostic phase. A holistic approach is recommended in caring for patients with SLE.


Human cell responses to ionizing radiation are differentially affected by the expressed connexins.
In multicellular organisms, intercellular communication is essential for homeostatic functions and has a major role in tissue responses to stress. Here, we describe the effects of expression of different connexins, which form gap junction channels with different permeabilities, on the responses of human cells to ionizing radiation. Exposure of confluent HeLa cell cultures to (137)Cs γ rays, 3.7 MeV α particles, 1000 MeV protons or 1000 MeV/u iron ions resulted in distinct effects when the cells expressed gap junction channels composed of either connexin26 (Cx26) or connexin32 (Cx32). Irradiated HeLa cells expressing Cx26 generally showed decreased clonogenic survival and reduced metabolic activity relative to parental cells lacking gap junction communication. In contrast, irradiated HeLa cells expressing Cx32 generally showed enhanced survival and greater metabolic activity relative to the control cells. The effects on clonogenic survival correlated more strongly with effects on metabolic activity than with DNA damage as assessed by micronucleus formation. The data also showed that the ability of a connexin to affect clonogenic survival following ionizing radiation can depend on the specific type of radiation. Together, these findings show that specific types of connexin channels are targets that may be exploited to enhance radiotherapeutic efficacy and to formulate countermeasures to the harmful effects of specific types of ionizing radiation.


Changes in transfusion practice over time in adult patients undergoing liver transplantation.
The aim of this study was to investigate changes in transfusion practice over time in liver transplantation surgery and to evaluate potential causes for changes in practice and report associated perioperative morbidity and mortality. A retrospective cohort study. A single tertiary referral academic hospital. Two cohorts of 100 sequential adult primary liver transplant recipients: Early practice (1990-1991) and recent practice (2005-2006). None. Perioperative transfusion and hemoglobin data were recorded. Mortality and postoperative complications were identified up to 30 days postoperatively. Appropriate intergroup statistical comparisons were made; p ≤ 0.05 was considered statistically significant. Compared with the early group, the recent group had significantly fewer perioperative allogeneic red blood cell transfusions, intraoperative autotransfusions, and transfusions of other blood products. No change in perioperative transfusion triggers was identified. There were no significant alterations in perioperative morbidity or mortality. When compared with patients in the early group, recent cohort patients received significantly fewer blood transfusions. The authors attribute this observation to changes in surgical technique rather than a significant alteration in transfusion triggers over the studied time period.


Neural correlates of error prediction in a complex motor task.
The goal of the study was to quantify error prediction processes via neural correlates in the Electroencephalogram (EEG). Access to such a neural signal will allow to gain insights into functional and temporal aspects of error perception in the course of learning. We focused on the error negativity (Ne) or error-related negativity (ERN) as a candidate index for the prediction processes. We have used a virtual goal-oriented throwing task where participants used a lever to throw a virtual ball displayed on a computer monitor with the goal of hitting a virtual target as often as possible. After one day of practice with 400 trials, participants performed another 400 trials on a second day with EEG measurement. After error trials (i.e., when the ball missed the target), we found a sharp negative deflection in the EEG peaking 250 ms after ball release (mean amplitude: t = -2.5, df = 20, p = 0.02) and another broader negative deflection following the first, reaching from about 300 ms after release until unambiguous visual knowledge of results (KR; hitting or passing by the target; mean amplitude: t = -7.5, df = 20, p < 0.001). According to shape and timing of the two deflections, we assume that the first deflection represents a predictive Ne/ERN (prediction based on efferent commands and proprioceptive feedback) while the second deflection might have arisen from action monitoring.


Experimental approaches to the human renal transcriptome.
The sum of RNA transcripts of a cell, organ structure, or organism can be referred to as transcriptome. An increasing number of studies report on specific and common alterations in the renal transcriptome in human nephropathies. In this review several challenges in transcriptomic analyses of the human kidney are discussed. This includes ways to approach the heterogeneity of the kidney itself as well as the diversity of renal diseases. Conventional and upcoming techniques for transcriptional profiling of minute tissue samples are presented, including so-called next generation sequencing and microRNA detection. Different tools to integrate transcriptomic data in a systematic context are discussed beside the current challenge to combine such results with data sets from other integrative biology technologies.


High turnover of fungal hyphae in incubation experiments.
Soil biological studies are often conducted on sieved soils without the presence of plants. However, soil fungi build delicate mycelial networks, often symbiotically associated with plant roots (mycorrhizal fungi). We hypothesized that as a result of sieving and incubating without plants, the total fungal biomass decreases. To test this, we conducted three incubation experiments. We expected total and arbuscular mycorrhizal (AM) fungal biomass to be higher in less fertilized soils than in fertilized soils, and thus to decrease more during incubation. Indeed, we found that fungal biomass decreased rapidly in the less fertilized soils. A shift towards thicker hyphae occurred, and the fraction of septate hyphae increased. However, analyses of phospholipid fatty acids (PLFAs) and neutral lipid fatty acids could not clarify which fungal groups were decreasing. We propose that in our soils, there was a fraction of fungal biomass that was sensitive to fertilization and disturbance (sieving, followed by incubation without plants) with a very high turnover (possibly composed of fine hyphae of AM and saprotrophic fungi), and a fraction that was much less vulnerable with a low turnover (composed of saprotrophic fungi and runner hyphae of AMF). Furthermore, PLFAs might not be as sensitive in detecting changes in fungal biomass as previously thought.


Shared risk factors for mood-, eating-, and weight-related health outcomes.
Given the overlap among depressive symptoms, disordered eating, and overweight, identifying shared risk factors for these conditions may inform public health interventions. This study aimed to examine cross-sectional and prospective relationships among these 3 conditions, and identify potential shared eating-related and psychosocial variable risk factors (i.e., body dissatisfaction, dieting, teasing experiences). A population-based sample (n = 1,902) self-reported depressive symptoms, disordered eating (binge eating, extreme weight control behaviors), weight status, and several putative risk factors (body satisfaction, dieting frequency, weight-related teasing) at 5-year intervals spanning early/middle adolescence, middle adolescence/early young adulthood, and early/middle young adulthood. There was moderate overlap among depressive symptoms, disordered eating, and overweight at each time point, and moderate stability in each condition over time. Body dissatisfaction and dieting were the most potent shared risk factors for later depressive symptoms, disordered eating, and overweight among males and females (ps < .05). Depressive symptoms, disordered eating, and overweight share several risk factors, including dieting and body dissatisfaction, which may be effective targets for interventions aiming to simultaneously prevent these 3 conditions.


Michigan's fee-for-value physician incentive program reduces spending and improves quality in primary care.
As policy makers and others seek to reduce health care cost growth while improving health care quality, one approach gaining momentum is fee-for-value reimbursement. This payment strategy maintains the traditional fee-for-service arrangement but includes quality and spending incentives. We examined Blue Cross Blue Shield of Michigan's Physician Group Incentive Program, which uses a fee-for-value approach focused on primary care physicians. We analyzed the program's impact on quality and spending from 2008 to 2011 for over three million beneficiaries in over 11,000 physician practices. Participation in the incentive program was associated with approximately 1.1 percent lower total spending for adults (5.1 percent lower for children) and the same or improved performance on eleven of fourteen quality measures over time. Our findings contribute to the growing body of evidence about the potential effectiveness of models that align payment with cost and quality performance, and they demonstrate that it is possible to transform reimbursement within a fee-for-service framework to encourage and incentivize physicians to provide high-quality care, while also reducing costs.


Cerebrospinal fluid beta-amyloid1-42 and tau in control subjects at risk for Alzheimer's disease: the effect of APOE epsilon4 allele.
Cerebrospinal fluid (CSF) measures of beta-amyloid(1-42) and tau are linked with the known neuropathology of Alzheimer's disease (AD). Numerous lines of evidence have also suggested that individuals with at least one APOE epsilon4 allele on chromosome 19 are at increased risk of developing AD. We tested these CSF markers in groups of subjects with AD and healthy older control subjects, using the absence or presence of the APOE epsilon4 allele as a predictive variable in the search for possible prognostic biomarkers of AD. We assessed the levels of beta-amyloid(1-42) and total tau in the CSF of 292 subjects (142 control subjects and 150 subjects with mild-to-moderate AD), who were research participants at the National Institute of Mental Health. The group of control subjects was enriched with a high percentage of subjects with a positive family history of AD. All subjects underwent extensive global cognitive testing. When divided according to the absence or presence of the APOE epsilon4 allele, the control subjects with at least one epsilon4 allele had significantly lower CSF beta-amyloid(1-42) but not tau levels than control subjects without an APOE epsilon4 allele (p < .01). As expected, the AD patients had lower levels of CSF beta-amyloid(1-42) and higher CSF tau levels than the normal control group (p < .01). The association of APOE epsilon4 allele and lower, more AD-like levels of CSF beta-amyloid(1-42) in older control subjects is consistent with previous studies showing possible neuroimaging and cognitive abnormalities with epsilon4 carriers and suggests that CSF beta-amyloid(1-42) decreases might represent an early biomarker of AD. Longitudinal follow-up is of course required to verify whether this biomarker is indeed predictive of clinical conversion to AD.


Human melanopsin forms a pigment maximally sensitive to blue light (λmax ≈ 479 nm) supporting activation of G(q/11) and G(i/o) signalling cascades.
A subset of mammalian retinal ganglion cells expresses an opsin photopigment (melanopsin, Opn4) and is intrinsically photosensitive. The human retina contains melanopsin, but the literature lacks a direct investigation of its spectral sensitivity or G-protein selectivity. Here, we address this deficit by studying physiological responses driven by human melanopsin under heterologous expression in HEK293 cells. Luminescent reporters for common second messenger systems revealed that light induces a high amplitude increase in intracellular calcium and a modest reduction in cAMP in cells expressing human melanopsin, implying that this pigment is able to drive responses via both Gq and Gi/o class G-proteins. Melanopsins from mouse and amphioxus had a similar profile of G-protein coupling in HEK293 cells, but chicken Opn4m and Opn4x pigments exhibited some Gs activity in addition to a strong Gq/11 response. An action spectrum for the calcium response in cells expressing human melanopsin had the predicted form for an opsin : vitamin A1 pigment and peaked at 479 nm. The G-protein selectivity and spectral sensitivity of human melanopsin is similar to that previously described for rodents, supporting the utility of such laboratory animals for developing methods of manipulating this system using light or pharmacological agents.


[Pathophysiology of ischaemia-reperfusion injury].
Reperfusion of ischaemic tissue provides oxygen and substrates that are necessary for tissue recovery and concurrently removes toxic metabolites. However, reperfusion may induce various detrimental processes that may cause further tissue damage. Such deterioration of tissue function after reperfusion is defined as ischaemia-reperfusion injury. The consequences of ischemia-reperfusion injury vary from reversible cell dysfunction to local and remote tissue destruction, multiple organ failure and death. The pathogenesis of ischaemia-reperfusion injury is complex and includes excessive production of reactive oxygen species, activation of neutrophils, activation of complement, involvement of cytokines and other inflammatory mediators, vasoactive substances NO and endothelin. This review discusses the pathophysiology of ischaemia-reperfusion injury, the mechanisms of reactive oxygen species production, and the role of other factors in the pathogenesis of such injury. Several approaches and procedures used in pre-clinical and clinical studies in order to limit ischaemia-reperfusion injury are also presented.


Validation of a centrifugal microfluidic sample lysis and homogenization platform for nucleic acid extraction with clinical samples.
The applications of microfluidic technologies in medical diagnostics continue to increase, particularly in the field of nucleic acid diagnostics. While much attention has been focused on the development of nucleic acid amplification and detection platforms, sample preparation is often taken for granted or ignored all together. Specifically, little or no consideration is paid to the development of microfluidic systems that efficiently extract nucleic acids from biological samples. Here, a centrifugal microfluidic platform for mechanical sample lysis and homogenization is presented. The system performs sample lysis through a magnetically actuated bead-beating system followed by a centrifugal clarification step. The supernatant is then transferred for extraction using a unique siphon. Several other new microfluidic functions are implemented on this centrifugal platform as well, including sample distribution, a unique hydraulic capillary valve, and self-venting. Additionally, the improved system has features with a small footprint designed specifically for integration with further downstream processing steps. Biological validation of the platform is performed using Bacillus subtilis spores and clinical samples (nasopharyngeal aspirates) for respiratory virus detection. The platform was found to be as efficient as in-tube bead-beating lysis and homogenization for nucleic acid extraction, and capable of processing 4 samples in batch to near PCR-ready products in under 6 min.


The consequences of heterogamy and homogamy on the similarity between spouses.
Humans in many societies are known to mate, or marry, assortatively for a number of characters such as eye colour, height, IQ and place of birth. In this assortment an element of active choice may be involved. It is not known whether this choice is genetic. Two models of human mate choice are examined in which both males and females can express a mating preference. In the first, 'sexual' preferences can be expressed for any phenotype not necessarily one's own; in the second, preferences are only expressed for an individual's own phenotype. The results of the examination indicate how much active choice would be needed to account for the observed correlations between human mates, and suggest whether human mating preferences are more likely to be sexual or assorting.


The efficacy of methotrexate for maintaining remission in inflammatory bowel disease.
The management of patients with inflammatory bowel disease who are resistant to or intolerant of azathioprine remains a challenge. Low-dose methotrexate has been shown to be effective in inducing remission in Crohn's disease. This review was conducted because there are limited long-term follow-up data during and after stopping treatment. There are also limited data on the use of methotrexate in ulcerative colitis. The study was a retrospective review of clinical notes. Remission was defined as minimal bowel symptoms without the need for oral steroids for 3 months. Relapse was defined as bowel symptoms that required steroid treatment or surgery. Seventy patients were reviewed; 48 had Crohn's disease and 22 had ulcerative colitis. The mean duration of treatment was 17.1 months; the mean maintenance dose was 20 mg weekly. Remission was achieved in 34 of 55 patients who completed more than 3 months of treatment (62%). Life-table analysis showed that the chances of remaining in remission at 12, 24 and 36 months (if treatment was continued) were 90%, 73% and 51%, respectively. The chances of remaining in remission after stopping treatment at 6, 12 and 18 months were 42%, 21% and 16%, respectively. The dose of methotrexate (mg/kg) was associated with the induction of remission (P=0.02). Treatment was equally effective for Crohn's disease and ulcerative colitis. Maintenance methotrexate treatment gives acceptable remission rates for treatment periods up to 3 years. After stopping treatment, relapse is frequent and occurs early (usually within 1 year).


Manifestations of early brain recovery associated with abstinence from alcoholism.
Chronic alcohol abuse results in morphological, metabolic, and functional brain damage which may, to some extent, be reversible with early effects upon abstinence. Although morphometric, spectroscopic, and neuropsychological indicators of cerebral regeneration have been described previously, the overall amount and spatial preference of early brain recovery attained by abstinence and its associations with other indicators of regeneration are not well established. We investigated global and local brain volume changes in a longitudinal two-timepoint study with T1-weighted MRI at admission and after short-term (6-7 weeks) sobriety follow-up in 15 uncomplicated, recently detoxified alcoholics. Volumetric brain gain was related to metabolic and neuropsychological recovery. On admission and after short-term abstinence, structural image evaluation using normalization of atrophy (SIENA), its voxelwise statistical extension to multiple subjects, proton MR spectroscopy (1H-MRS), and neuropsychological tests were applied. Upon short-term sobriety, 1H-MRS levels of cerebellar choline and frontomesial N-acetylaspartate (NAA) were significantly augmented. Automatically detected global brain volume gain amounted to nearly two per cent on average and was spatially significant around the superior vermis, perimesencephalic, periventricular and frontal brain edges. It correlated positively with the percentages of cerebellar and frontomesial choline increase, as detected by 1H-MRS. Moreover, frontomesial NAA gains were associated with improved performance on the d2-test of attention. In 10 age- and gender-matched healthy control subjects, no significant brain volume or metabolite changes were observed. Although cerebral osmotic regulations may occur initially upon sobriety, significant increases of cerebellar choline and frontomesial NAA levels detected at stable brain water integrals and creatine concentrations, serum electrolytes and red blood cell indices in our patient sample suggest that early brain recovery through abstinence does not simply reflect rehydration. Instead, even the adult human brain and particularly its white matter seems to possess genuine capabilities for regrowth. Our findings emphasize metabolic as well as regionally distinct morphological capacities for partial brain recovery from toxic insults of chronic alcoholism and substantiate early measurable benefits of therapeutic sobriety. Further understanding of the precise mechanisms of this recovery may become a valuable model of brain regeneration with relevance for other disorders.


Schizophrenia and the parvalbumin-containing class of cortical local circuit neurons.
The purpose of this study was to test the hypothesis that abnormalities in the parvalbumin-containing subclass of local circuit neurons contribute to altered gamma-aminobutyric acid (GABA) neurotransmission in the prefrontal cortex of schizophrenic subjects. Profile counts and somal size measures were made of parvalbumin-immunoreactive neurons in areas 9, 46, and 17 from 15 matched pairs of schizophrenic and normal comparison subjects. No differences in relative density, laminar distribution, or somal size of labeled neurons were found in any region. These findings suggest that altered GABA neurotransmission in schizophrenia is due to either abnormalities in other sub-populations of prefrontal cortical GABA neurons or abnormalities in the parvalbumin-containing subclass that could not be detected in the present study.