Highly fatal fast-channel syndrome caused by AChR ε subunit mutation at the agonist binding site.
To characterize the molecular basis of a novel fast-channel congenital myasthenic syndrome. We used the candidate gene approach to identify the pathogenic mutation in the acetylcholine receptor (AChR) ε subunit, genetically engineered the mutant AChR into HEK cells, and evaluated the level of expression and kinetic properties of the mutant receptor. An 8-year-old boy born to consanguineous parents had severe myasthenic symptoms since birth. He is wheelchair bound and pyridostigmine therapy enables him to take only a few steps. Three similarly affected siblings died in infancy. He carries a homozygous p.W55R mutation at the α/ε subunit interface of the AChR agonist binding site. The mutant protein expresses well in HEK cells. Patch-clamp analysis of the mutant receptor expressed in HEK cells reveals 30-fold reduced apparent agonist affinity, 75-fold reduced apparent gating efficiency, and strikingly attenuated channel opening probability (P(open)) over a range agonist concentrations. Introduction of a cationic Arg into the anionic environment of α/ε AChR binding site hinders stabilization of cationic ACh by aromatic residues and accounts for the markedly perturbed kinetic properties of the receptor. The very low P(open) explains the poor response to pyridostigmine and the high fatality of the disease.


Focused Ethnographic Examination of Barriers to Use of Long-Acting Injectable Antipsychotics.
The authors designed this project to identify barriers to using long-acting formulations of antipsychotics. The authors used a focused ethnographic approach. Patients, psychiatrists, nurses, therapists and administrators were interviewed about barriers to use of long-acting injectable (LAI) antipsychotics at six facilities in New York State, as were representatives from insurance firms, a pharmaceutical company, and a national professional organization. Interviews were conducted and analyzed by a central team not affiliated with the institutions. Interviews were obtained with 23 patients, 16 psychiatrists, three nurses, 23 therapists, 14 administrators, four insurers, one representative from a pharmaceutical industry, and one representative from a national organization. Major barriers identified from the interviews included restricting discussions about LAI medication to only patients with nonadherence or repeated hospitalizations; inadequate education efforts with patients about LAI antipsychotics; inadequate support for patients making medication decisions; lack of communication within the treatment team about issues relevant to use of LAI formulations by patients; therapists' limited knowledge about LAI antipsychotics, which restricted their role in supporting patients making treatment decisions; psychiatrist concerns about the pharmacologic properties of LAI formulations; lack of clinic infrastructure to support LAI prescriptions; and payer concerns about whether the immediate costs of LAI administration would translate into later potential cost benefits. Effective shared decision making about use of LAI antipsychotics requires that patients receive accurate information and support for their decision making. The training needs and administrative support requirements of all team members should be considered to provide patients with the information and support required.


Secretoneurin--a neuropeptide generated in brain, adrenal medulla and other endocrine tissues by proteolytic processing of secretogranin II (chromogranin C).
Secretogranin II (chromogranin C), originally described as tyrosine sulfated protein of the anterior pituitary, is present in large dense core vesicles of several endocrine cells and neurons. We raised antisera in rabbits to conjugates of two synthetic peptides (bovine secretogranin 133-151 and rat secretogranin 154-186) flanked in the primary structure of secretogranin II by pairs of basic residues and used them to investigate the proteolytic processing of this protein by immunoblotting and a newly developed radioimmunoassay. The sensitivity of this assay was 30 fmol for secretogranin 154-186 and 60 fmol for secretogranin 133-151. The highest degree of processing of secretogranin II (> 90%) occurs in brain. One of the peptides (secretogranin 133-151) is not generated to any significant extent. The other peptide, secretogranin 154-186, however, is formed in vivo, and in brain the free peptide apparently represents the predominant form. The highest concentrations of secretogranin 154-186 are found in the hypothalamus, two- to six-fold lower levels are present in the hippocampus, caudate nucleus, thalamus and brainstem. These concentrations are comparable to those of established neuropeptides. In order to indicate the special relevance of secretogranin II and of this peptide for brain we have named this peptide secretoneurin. The newly developed radioimmunoassay for this peptide will be a useful tool to establish its physiologic role in brain.


CDC4 mutations occur in a subset of colorectal cancers but are not predicted to cause loss of function and are not associated with chromosomal instability.
CDC4/FBXW7 is part of a ubiquitin ligase complex which targets molecules such as cyclin E, c-myc, and c-jun for destruction. CDC4 mutations occur in several cancer types and are best described in colorectal tumors. Knockout of CDC4 in vitro in colorectal cancer cells causes changes suggestive of chromosomal instability (CIN). In p53(+/-) mice, radiation-induced lymphomas show deletion or mutation of one copy of CDC4 and knockdown of CDC4 leads to increased aneuploidy in mouse fibroblasts. We screened 244 colorectal tumors and 40 cell lines for CDC4 mutations and allelic loss. Six percent (18 of 284) of tumors, including near-diploid (CIN-) lesions, harbored CDC4 mutations and there was no association between mutation and CIN (polyploidy). The CDC4 mutation spectrum in colorectal tumors was heavily biased towards C:G > T:A changes, either missense mutations at critical arginine residues or nonsense changes in the 5' half of the gene. The reasons for this odd mutation spectrum were unclear but C:G > T:A changes were not found more often than expected at APC, K-ras, or p53 in the same tumors and we found no specific defects in DNA repair to account for the observations. No colorectal tumor was found to carry two CDC4 mutations predicted to abolish protein function; partial loss of CDC4 function may therefore cause tumorigenesis. The in vitro studies, therefore, did not assess the functional effects of mutant alleles which are found in vivo. CDC4 mutations may be selected primarily to drive progression through the cell cycle although CIN might be an important secondary effect in some cancers.


Reducing prescribing errors: can a well-designed electronic system help?
In this study, the aim was to investigate if an electronic prescribing system designed specifically to reduce errors would lead to fewer errors in prescribing medicines in a secondary care setting. The electronic system was compared with paper prescription charts on 16 intensive care patients to assess any change in the number of prescribing errors. The overall level of compliance with nationally accepted standards was significantly higher with the electronic system (91.67%) compared with the paper system (46.73%). Electronically generated prescriptions were found to contain significantly fewer deviations (28 in 329 prescriptions, 8.5%) than the written prescriptions (208 in 408 prescriptions, 51%). Taking an interdisciplinary approach to work on the creation of a system designed to minimize the risk of error has resulted in a favoured system that significantly reduces the number of errors made.


hnRNP C1/C2 and Pur-beta proteins mediate induction of senescence by oligonucleotides homologous to the telomere overhang.
Experimental disruption of the telomere overhang induces a potent DNA damage response and is the target of newly emerging cancer therapeutics. Introduction of T-oligo, an eleven-base oligonucleotide homologous to the 3'-telomeric overhang, mimics telomere disruption and induces DNA damage responses through activation of p53, p73, p95/Nbs1, E2F1, pRb, and other DNA damage response proteins. ATM (ataxia telangiectasia mutated) was once thought to be the primary driver of T-oligo-induced DNA damage responses; however, recent experiments have highlighted other key proteins that may also play a significant role. To identify proteins associated with T-oligo, MM-AN cells were treated with biotinylated T-oligo or complementary oligonucleotide, cell lysates were run on SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and the protein bands observed after treatment of cells with T-oligo or complementary oligonucleotide were analyzed using mass spectrometry. To study the effect of T-oligo on expression of hnRNP C1/C2 (heterogeneous nuclear ribonucleoprotein C1 and C2) and purine-rich element binding proteins (Pur proteins), cells were treated with T-oligo, and immunoblotting experiments were performed. To determine their role in senescence, cells were treated with shRNA (short hairpin ribonucleic acid) against these proteins, and senescence was studied using the senescence associated beta-galactosidase assay. Using mass spectrometry, RNA-binding hnRNP C1/C2 and DNA-binding Pur proteins were found to associate with T-oligo. hnRNP C1/C2 exhibited increased expression (3.6-12.0-fold) in non-small-cell lung cancer (NSCLC) and in melanoma cells (4.5-5.2-fold), and Pur proteins exhibited increased expression of 2.2-fold in NSCLC and 2.0-fold in melanoma cells after T-oligo treatment. Experimental knockdown of hnRNP C1/C2 and Pur-beta completely abrogated T-oligo induced senescence in both MU melanoma and H358 NSCLC cells. Additionally, knockdown of Pur-beta prevented T-oligo-induced phosphorylation of p53, hypophosphorylation of pRb, and upregulation of E2F1, p21, and p53. These novel findings highlight proteins essential to T-oligo's anticancer effects that may be of interest in telomere biology and cancer therapeutics.


Development and testing of coatings for orbital space radiation environments.
Specific coating processes and materials were investigated in the quest to develop multilayer coatings with greater tolerance to space radiation exposure. Ultraviolet reflection (UVR) and wide-band antireflection (AR) multilayer coatings were deposited on solar cell covers and test substrates and subsequently exposed to simulated space environments and also flown on the Materials International Space Station Experiment-7 (MISSE-7) to determine their space environment stability. Functional solar cells integrated with these coatings underwent simulated UV and MISSE-7 low earth orbit flight exposure. The effects of UV, proton, and atomic oxygen exposure on coatings and on assembled solar cells as related to the implemented deposition processes and material compositions were small. The UVR/AR coatings protected flexible polymer substrate materials that are intended for future flexible multijunction cell arrays to be deployed from rolls. Progress was made toward developing stable and protective coatings for extended space-mission applications. Test results are presented.


Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase.
Characterization of the proteome of organelles and subcellular domains is essential for understanding cellular organization and identifying protein complexes as well as networks of protein interactions. We established a proteomic mapping platform in live Drosophila tissues using an engineered ascorbate peroxidase (APEX). Upon activation, the APEX enzyme catalyzes the biotinylation of neighboring endogenous proteins that can then be isolated and identified by mass spectrometry. We demonstrate that APEX labeling functions effectively in multiple fly tissues for different subcellular compartments and maps the mitochondrial matrix proteome of Drosophila muscle to demonstrate the power of APEX for characterizing subcellular proteomes in live cells. Further, we generate "MitoMax," a database that provides an inventory of Drosophila mitochondrial proteins with subcompartmental annotation. Altogether, APEX labeling in live Drosophila tissues provides an opportunity to characterize the organelle proteome of specific cell types in different physiological conditions.


A dynamic model of nutrient uptake by root hairs.
Root hairs are known to be important in the uptake of sparingly soluble nutrients by plants, but quantitative understanding of their role in this is weak. This limits, for example, the breeding of more nutrient-efficient crop genotypes. We developed a mathematical model of nutrient transport and uptake in the root hair zone of single roots growing in soil or solution culture. Accounting for root hair geometry explicitly, we derived effective equations for the cumulative effect of root hair surfaces on uptake using the method of homogenization. Analysis of the model shows that, depending on the morphological and physiological properties of the root hairs, one of three different effective models applies. They describe situations where: (1) a concentration gradient dynamically develops within the root hair zone; (2) the effect of root hair uptake is negligibly small; or (3) phosphate in the root hair zone is taken up instantaneously. Furthermore, we show that the influence of root hairs on rates of phosphate uptake is one order of magnitude greater in soil than solution culture. The model provides a basis for quantifying the importance of root hair morphological and physiological properties in overall uptake, in order to design and interpret experiments in different circumstances.


Increased Hs-CRP/adiponectin ratio is associated with increase carotid intima-media thickness.
High sensitivity C-reactive protein (Hs-CRP) and adiponectin (APN) are two critical cytokines and exert inverse effects on atherosclerosis initiation and progression. The purpose of our study was to investigate the value of Hs-CRP and ANP ratio (Hs-CRP/APN ratio) on evaluating atherosclerosis progression. One hundred sixty consecutive participants underwent carotid intima-media thickness (CIMT) measured by ultrasound were enrolled and drawn fasting blood samples for plasma levels Hs-CRP and APN, serum levels of lipid profiles and fasting blood glucose evaluation. Other anthropometrics and clinical status were collected by questionnaire. All participants were divided into 4 groups according to the baseline Hs-CRP/APN ratio and underwent CIMT measurement every 6 months. CIMT increment and composite cardiovascular endpoints were compared after 24 months' follow-up. At baseline, body mass index (BMI), smoking, diabetic mellitus, usage of statins, Hs-CRP and APN independently correlated with Hs-CRP/APN ratio as analyzed by spearman rank correlation. Smoking, serum level of LDL-C, plasma level of Hs-CRP and Hs-CRP/APN ratio were positively correlated with CIMT while usage of statins and plasma level of APN were negatively correlated with CIMT as analyzed by multiple linear regression analysis. After 24 months' follow-up, the progression of CIMT was the most prominent in the fourth quartile of baseline Hs-CRP/APN ratio. In addition, the incidence of composite cardiovascular endpoint was also higher in the fourth quartile as compared to the other 3 lower quartiles. Hs-CRP/APN ratio was a useful predictor to discriminate subjects who were at increased risk of atherosclerosis progression.


Gliding mutants of Myxococcus xanthus with high reversal frequencies and small displacements.
Myxococcus xanthus cells move on a solid surface by gliding motility. Several genes required for gliding motility have been identified, including those of the A- and S-motility systems as well as the mgl and frz genes. However, the cellular defects in gliding movement in many of these mutants were unknown. We conducted quantitative, high-resolution single-cell motility assays and found that mutants defective in mglAB or in cglB, an A-motility gene, reversed the direction of gliding at frequencies which were more than 1 order of magnitude higher than that of wild type cells (2.9 min-1 for DeltamglAB mutants and 2.7 min-1 for cglB mutants, compared to 0.17 min-1 for wild-type cells). The average gliding speed of DeltamglAB mutant cells was 40% of that of wild-type cells (on average 1.9 micrometers/min for DeltamglAB mutants, compared to 4.4 micrometers/min for wild-type cells). The mglA-dependent reversals and gliding speeds were dependent on the level of intracellular MglA protein: mglB mutant cells, which contain only 15 to 20% of the wild-type level of MglA protein, glided with an average reversal frequency of about 1.8 min-1 and an average speed of 2.6 micrometers/min. These values range between those exhibited by wild-type cells and by DeltamglAB mutant cells. Epistasis analysis of frz mutants, which are defective in aggregation and in single-cell reversals, showed that a frzD mutation, but not a frzE mutation, partially suppressed the mglA phenotype. In contrast to mgl mutants, cglB mutant cells were able to move with wild-type speeds only when in close proximity to each other. However, under those conditions, these mutant cells were found to glide less often with those speeds. By analyzing double mutants, the high reversing movements and gliding speeds of cglB cells were found to be strictly dependent on type IV pili, encoded by S-motility genes, whereas the high-reversal pattern of mglAB cells was only partially reduced by a pilR mutation. These results suggest that the MglA protein is required for both control of reversal frequency and gliding speed and that in the absence of A motility, type IV pilus-dependent cell movement includes reversals at high frequency. Furthermore, mglAB mutants behave as if they were severely defective in A motility but only partially defective in S motility.


Leg strength declines with advancing age despite habitual endurance exercise in active older adults.
Age-associated loss of muscle mass (sarcopenia) and strength (dynapenia) is associated with a loss of independence that contributes to falls, fractures, and nursing home admissions, whereas regular physical activity has been suggested to offset these losses. The purpose of this study was to evaluate the effect of habitual endurance exercise on muscle mass and strength in active older adults. A longitudinal analysis of muscle strength (≈4.8 years apart) was performed on 59 men (age at start of study: 58.6 ± 7.3 years) and 35 women (56.9 ± 8.2 years) who used endurance running as their primary mode of exercise. There were no changes in fat-free mass although body fat increased minimally (1.0-1.5%). Training volume (km·wk, d·wk) decreased in both the men and women. There was a significant loss of both isometric knee extension (≈5% per year) and knee flexion (≈3.6% per year) strength in both the men and women. However, there was no significant change in either isokinetic concentric or eccentric torque of the knee extensors. Our data demonstrated a significant decline in isometric knee extensor and knee flexor strength although there were no changes in body mass in this group of very active older men and women. Our data support newer exercise guidelines for older Americans suggesting resistance training be an integral component of a fitness program and that running alone was not sufficient to prevent the loss in muscle strength (dynapenia) with aging.


Phage-Antibiotic Synergy via Delayed Lysis.
When phages infect bacteria cultured in the presence of sublethal doses of antibiotics, the sizes of the phage plaques are significantly increased. This phenomenon is known as phage-antibiotic synergy (PAS). In this study, the observation of PAS was extended to a wide variety of bacterium-phage pairs using different classes of antibiotics. PAS was shown in both Gram-positive and Gram-negative bacteria. Cells stressed with β-lactam antibiotics filamented or swelled extensively, resulting in an increase in phage production. PAS was also sometimes observed in the presence of other classes of antibiotics with or without bacterial filamentation. The addition of antibiotics induced recA expression in various bacteria, but a recA deletion mutant strain of Escherichia coli also showed filamentation and PAS in the presence of quinolone antibiotics. The phage adsorption efficiency did not change in the presence of the antibiotics when the cell surfaces were enlarged as they filamented. Increases in the production of phage DNA and mRNAs encoding phage proteins were observed in these cells, with only a limited increase in protein production. The data suggest that PAS is the product of a prolonged period of particle assembly due to delayed lysis. The increase in the cell surface area far exceeded the increase in phage holin production in the filamented host cells, leading to a relatively limited availability of intracellular holins for aggregating and forming holes in the host membrane. Reactive oxygen species (ROS) stress also led to an increased production of phages, while heat stress resulted in only a limited increase in phage production.IMPORTANCE Phage-antibiotic synergy (PAS) has been reported for a decade, but the underlying mechanism has never been vigorously investigated. This study shows the presence of PAS from a variety of phage-bacterium-antibiotic pairings. We show that increased phage production resulted directly from a lysis delay caused by the relative shortage of holin in filamented bacterial hosts in the presence of sublethal concentrations of stress-inducing substances, such as antibiotics and reactive oxygen species (ROS).


Testing an empowerment intervention to help parents make homes smoke-free: a randomized controlled trial.
The objective of this randomized controlled trial was to test if parents' participation in an intervention based on an empowerment ideology and participatory experiences decreased the number of cigarettes smoked in homes. Sixty families were randomized to the intervention (n = 30) or control (n = 30) group. The intervention included three weekly group sessions followed by three weekly follow-up telephone calls over six consecutive weeks. During group sessions, parents shared experiences about environmental tobacco smoke, identified personal strengths and resources, and developed action plans. Data were collected in interviewer-administered questionnaires at baseline and 6 months follow-up. Ninety-three percent of the sample consisted of mothers, 77% of whom smoked during pregnancy. Forty-two percent of the total sample reported a household income of <$15,000. The median number of cigarettes smoked in the home daily decreased from 18 to 4 in the total sample however no statistically significant difference was detected between groups at 6 months follow-up. Participation in the study, independent of group, may have resulted in parents decreasing the number of cigarettes smoked in the home. Valuable lessons were learned about recruiting and working with this group of parents, all of whom faced the challenges of tobacco and almost half of whom lived in poverty.


Lyophilized Amorphous Dispersion of Telmisartan in a Combined Carrier-Alkalizer System: Formulation Development and In Vivo Study.
Telmisartan suffers from low oral bioavailability due to its poor water solubility. The research work presents a formulation of solid dispersed (SD) telmisartan formulation as a ternary mixture of a drug, a polymeric carrier (poly(vinylpyrrolidone) (PVP) K30), and an alkalizer (Na2CO3). The preparation method, which was lyophilization of an aqueous solution containing the ingredients, was free from any organic solvent. The developed SD formulations resulted in a significant improvement in in vitro dissolution (>90% drug dissolution in 15 min) compared to pure telmisartan. Solid-state characterization by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and X-ray diffraction (XRD) studies indicated the conversion of crystalline telmisartan into an amorphous form. Fourier transform infrared (FTIR) spectroscopy revealed the drug-polymer interaction that was responsible for reducing the chances of recrystallization. A short-term stability study showed that selected SD formulations were stable in terms of in vitro dissolution and retained their amorphous structure in ambient and accelerated conditions over 2 months. Selected formulations (drug/PVP K30/Na2CO3 as 1:1:2 or 1:2:2 weight ratio) resulted in >2.48 times relative oral bioavailability compared to marketed formulations. It was considered that the incorporation of an alkalizer and a hydrophilic polymer, and amorphization of telmisartan by lyophilization, could enhance in vitro dissolution and improve oral bioavailability.


Associations between migration experience and perceived mental health in optimal ageing: Evidence from the Sardinian Blue Zone.
The effect of migration on perceived mental health has not been examined in older migrants after their return to their places of origin. This study was mainly aimed at evaluating the perceived mental health of older people who experienced migration and permanent resident peers living in the Sardinian Blue Zone (i.e. one of the four areas of exceptional longevity in the world). Forty-eight community-based older participants (32 males and 16 females) with and without a migration experience were recruited in two villages of the Sardinian Blue Zone and completed a battery of self-report inventories assessing psychological well-being, negative mood, and ego resilience. Older individuals who experienced migration reported higher ego resilience and exhibited greater resources used to manage positive emotionality (i.e. openness to life experiences). Moreover, compared to the normative data, both the groups reported higher psychological well-being and fewer depressive symptoms. Finally, no significant associations were found between the length of migration and each mental health index. In conclusion, resilience seems to represent a psychological trait that helps to manage stressful events and contributes to the preservation of perceived mental health in late adulthood.


Antistress Effects of Rosa rugosa Thunb. on Total Sleep Deprivation-Induced Anxiety-Like Behavior and Cognitive Dysfunction in Rat: Possible Mechanism of Action of 5-HT6 Receptor Antagonist.
Our previous results suggest that the Rosa rugosa Thunb. (family Rosaceae) alleviates endurance exercise-induced stress by decreasing oxidative stress levels. This study aimed to screen and identify the physiological antistress effects of an extract of R. rugosa (RO) on sleep deprivation-induced anxiety-like behavior and cognitive tests (in vivo) and tested for hippocampal CORT and monoamine levels (ex vivo), corticosterone (CORT)-induced injury, N-methyl-d-aspartate (NMDA) receptor, and serotonin 6 (5-hydroxytryptamine 6, 5-HT6) receptor activities (in vitro) in search of active principles and underlying mechanisms of action. We confirmed the antistress effects of RO in a sleep-deprived stress model in rat and explored the underlying mechanisms of its action. In conclusion, an R. rugosa extract showed efficacy and potential for use as an antistress therapy to treat sleep deprivation through its antagonism of the 5-HT6 receptor and resulting inhibition of cAMP activity.


A comparison of concentrations of lead in human tissues.
This postmortem study of lead concentrations in the tissues of 129 subjects is an extension to a report by Barry and Mossman (1970). Lead concentrations in bone greatly exceeded the concentrations in soft tissues and were highest in the dense bones. Bone lead concentrations increased with age in both sexes, more especially in male subjects and in dense bone, varying between mean values of 2-16 ppm in the ribs of children to over 50 ppm in the dense petrous temporal bones of elderly male adults. Male adults contained over 30% more lead in their bones than females. Mean concentrations of lead in the soft tissues varied from less than 0-1 ppm in organs such as muscle and heart to over 2 ppm in the aorta. In most tissues with lead values in excess of 0-2 ppm the male concentrations exceeded female values by about 30%. With the exception of the aorta, spleen, lung, and prostate, lead concentrations did not increase with age in the soft tissues of either sex after about the second decade of life. Children showed concentrations of lead in their soft tissues comparable to female adults, but the concentrations in bone were much lower. It is suggested that children do not possess the same capacity as adults to retain lead in bone. In male adults occupationally exposed to lead the concentrations of lead in bone exceeded the concentrations in unexposed male adults within the same age group by two-to three-fold. Soft tissue lead concentrations between the two groups were less divergent. An assessment of the total body burden of lead revealed higher levels in adult male subjects than in females at mean values of 164-8 mg compared to 103-6 mg, respectively. Over 90% of the total body burden of lead in adults was in bone, of which over 70% was in dense bone. Male adults occupationally exposed to lead had mean total body burdens of 566-4 mg Pb, of which 97% was in bone. The release of lead from bone in conjunction with calcium was not considered to be of physiological significance. Lead concentrations of hair and nails were higher than soft tissue lead concentrations and varied widely. Hair lead measurements were not considered to provide a reliable assessment of lead absorption. The concentrations of lead in tissues of a mixed group of subjects with no known occupational exposure to lead have been shown to be comparable to the findings in earlier studies. Present levels of lead in the environment are not considered to be a hazard to the health of the population in general.


Evolution of tolerance in an invasive weed after reassociation with its specialist herbivore.
The interaction between the European wild parsnip Pastinaca sativa and its coevolved florivore the parsnip webworm Depressaria pastinacella, established in North America for over 150 years, has resulted in evolution of local chemical phenotype matching. The recent invasion of New Zealand by webworms, exposing parsnips there to florivore selection for the first time, provided an opportunity to assess rates of adaptive response in a real-time experiment. We planted reciprocal common gardens in the USA and NZ with seeds from (1) US populations with a long history of webworm association; (2) NZ populations that had never been infested and (3) NZ populations infested for 3 years (since 2007) or 6 years (since 2004). We measured impacts of florivory on realized fitness, reproductive effort and pollination success and measured phenotypic changes in infested NZ populations relative to uninfested NZ populations to determine whether rapid adaptive evolution in response to florivory occurred. Irrespective of country of origin or location, webworms significantly reduced plant fitness. Webworms reduced pollination success in small plants but not in larger plants. Although defence chemistry remained unchanged, plants in infested populations were larger after 3-6 years of webworm florivory. As plant size is a strong predictor of realized fitness, evolution of large size as a component of florivore tolerance may occur more rapidly than evolution of enhanced chemical defence.


Intraperitoneal hyaluronan administration in conscious rats: absorption, metabolism, and effects on peritoneal fluid dynamics.
Hyaluronan (HA) is a major component of interstitial tissue that participates in fluid homeostasis, response to inflammation, and wound healing. Previous studies have shown that intraperitoneal administration of HA can affect peritoneal fluid transport during short peritoneal dialysis exchanges in anesthetized rats. We sought to investigate the effect of high molecular weight HA on peritoneal permeability in conscious rats during dialysis exchanges up to 8 hours in duration. In addition, we sought to investigate the absorption of HA from the peritoneal cavity, its accumulation in peritoneal tissues, and its metabolism in normal and uremic rats. Experiments were performed on male Wistar rats infused with 30 mL peritoneal dialysis solution (Dianeal, Baxter Healthcare; Castelbar, Ireland) containing 10 mg/dL HA or with Dianeal alone (control). Peritoneal fluid removal (net ultrafiltration), permeability to glucose, creatinine, and total proteins, and tissue and blood levels of HA were determined in separate groups of rats at 1,2, 4, 6, and 8 hours after intraperitoneal infusion. Hyaluronan appearance and disappearance from plasma were also studied for 24 hours in separate groups of normal and uremic rats. Net ultrafiltration was significantly greater (27%) in rats infused with HA at 4, 6, and 8 hours (p < 0.01) compared to controls. Transperitoneal equilibration of protein was reduced by 27% (p < 0.001) at 4 hours and by 30% (p < 0.01) at 8 hours. During the 8-hour exchange, peritoneal clearance of creatinine increased by 27% (p < 0.01), whereas the clearance of total protein decreased by 27% (p < 0.005). After 8 hours, 25.7% +/- 3.1% of the administered HA was absorbed from the peritoneal cavity, peritoneal tissue HA concentration was increased by 117% (p < 0.001), and plasma HA levels increased by 435% (p < 0.001). Plasma HA levels returned to normal within 24 hours after intraperitoneal administration in both healthy and uremic rats. Hyaluronan added to dialysis fluid is absorbed from the peritoneal cavity and accumulates in peritoneal tissues. Hyaluronan supplementation produces changes in peritoneal permeability, leading to higher net ultrafiltration and peritoneal creatinine clearance, whereas total protein clearance decreases. The HA that is absorbed from the peritoneal cavity appears to be rapidly metabolized in both healthy and uremic rats.


Transversus thoracis muscle plane block in cardiac surgery: a pilot feasibility study.
Cardiac surgery patients often experience significant pain after median sternotomy. The transversus thoracis muscle plane (TTP) block is a newly developed, single-shot nerve block technique that provides analgesia for the anterior chest wall. In this double-blind pilot study, we assessed the feasibility of performing this novel block as an analgesic adjunct. All patients aged 18-90 undergoing elective cardiac surgery were randomized to the block or standard care control group on admission to the intensive care unit after surgery. Under ultrasound guidance, patients in the block group received the TTP block with 20 mL of either 0.3% or 0.5% ropivacaine bilaterally, based on weight. The control group did not receive any injections. All blocks were performed by a single anesthesiologist, and data collection was performed by blinded assessors. The primary feasibility outcomes were rate of recruitment, adherence, and adverse events. The rate of recruitment was defined as the ratio of patients giving informed consent to the number of eligible patients who were approached to participate. Secondary outcomes included 12-hour and 24-hour Numeric Rating Scale (NRS) pain scores, 24-hour hydromorphone and acetaminophen requirements, time to extubation, time to first opioid administration, and patient satisfaction (on a yes/no questionnaire) at 24 hours. Twenty patients were approached for this study and 19 were enrolled. Eight patients received the intended intervention in each group. The recruitment rate was 95% of all approached eligible patients, and the adherence rate to treatment group was 94%. There were no block-related adverse events. The mean (SD) NRS pain scores at rest were 3.3 (3.2) in the block group vs 5.6 (3.2) in the control group at 12 hours. At 24 hours, the pain scores were 4.1 (3.9) vs 4.1 (3.3) in the block and control group, respectively. The mean (SD) 24-hour hydromorphone administration was 1.9 (1.1) mg in the block group vs 1.8 (0.9) mg in the control group. The TTP block is a novel pain management strategy poststernotomy. The results reveal a high patient recruitment, adherence, and satisfaction rate, and provide some preliminary data supporting safety. NCT03128346.


Flare-productive active regions.
Strong solar flares and coronal mass ejections, here defined not only as the bursts of electromagnetic radiation but as the entire process in which magnetic energy is released through magnetic reconnection and plasma instability, emanate from active regions (ARs) in which high magnetic non-potentiality resides in a wide variety of forms. This review focuses on the formation and evolution of flare-productive ARs from both observational and theoretical points of view. Starting from a general introduction of the genesis of ARs and solar flares, we give an overview of the key observational features during the long-term evolution in the pre-flare state, the rapid changes in the magnetic field associated with the flare occurrence, and the physical mechanisms behind these phenomena. Our picture of flare-productive ARs is summarized as follows: subject to the turbulent convection, the rising magnetic flux in the interior deforms into a complex structure and gains high non-potentiality; as the flux appears on the surface, an AR with large free magnetic energy and helicity is built, which is represented by δ -sunspots, sheared polarity inversion lines, magnetic flux ropes, etc; the flare occurs when sufficient magnetic energy has accumulated, and the drastic coronal evolution affects magnetic fields even in the photosphere. We show that the improvement of observational instruments and modeling capabilities has significantly advanced our understanding in the last decades. Finally, we discuss the outstanding issues and future perspective and further broaden our scope to the possible applications of our knowledge to space-weather forecasting, extreme events in history, and corresponding stellar activities.


Alcohol use disorders after bariatric surgery.
The increased incidence of alcohol use disorders (AUD) after bariatric surgery has been proposed despite limited empirical support. We sought to determine the prevalence of current and lifetime AUD and other Axis I diagnoses in patients who have undergone bariatric surgery, and to test the hypothesis that greater weight loss is associated with a higher incidence of AUD following surgery. Individuals who underwent bariatric surgery between 2004 and 2007 were recruited for inclusion in the study. The diagnosis of current and lifetime AUD and other Axis I disorders was assessed using the Structured Clinical Interview for DSM-IV. A total of 51 individuals were included. The prevalence of lifetime and current AUD was 35.3% and 11.8%, respectively. No associations were found between weight loss following surgery and the development of an AUD or other Axis I diagnoses. Significantly more current AUD was reported in (1) individuals with a lifetime history of AUD compared to those without a lifetime AUD (p < 0.05), and (2) individuals undergoing Roux-en-Y gastric bypass (RYGB) compared to those undergoing the laparoscopic adjustable gastric banding (LAGB) surgery (p < 0.05). Individuals undergoing bariatric surgery were found to have a lifetime prevalence of AUD comparable to the general population. Although weight loss was not associated with the development of an AUD following surgery, individuals with a lifetime history of AUD may be at increased risk for relapsing to alcohol use after surgery. All instances of current AUD were identified in individuals undergoing RYGB as opposed to LAGB.


Grasses use an alternatively wired bHLH transcription factor network to establish stomatal identity.
Stomata, epidermal valves facilitating plant-atmosphere gas exchange, represent a powerful model for understanding cell fate and pattern in plants. Core basic helix-loop-helix (bHLH) transcription factors regulating stomatal development were identified in Arabidopsis, but this dicot's developmental pattern and stomatal morphology represent only one of many possibilities in nature. Here, using unbiased forward genetic screens, followed by analysis of reporters and engineered mutants, we show that stomatal initiation in the grass Brachypodium distachyon uses orthologs of stomatal regulators known from Arabidopsis but that the function and behavior of individual genes, the relationships among genes, and the regulation of their protein products have diverged. Our results highlight ways in which a kernel of conserved genes may be alternatively wired to produce diversity in patterning and morphology and suggest that the stomatal transcription factor module is a prime target for breeding or genome modification to improve plant productivity.


Evaluation and the NIH clinical and translational science awards: a "top ten" list.
Since 2006, a total of 61 Clinical and Translational Science Institutes (CTSAs) have been funded by the National Institutes of Health (NIH), with the aim of reducing translation time from a bench discovery to when it impacts patients. This special issue of Evaluation & the Health Professions focuses on evaluation within and across the large, complex system of the CTSA Program of NIH. Through insights gained by reading the articles in this special edition and the experience of the authors, a "top ten" list of lessons learned and insights gained is presented. The list outlines issues that face those who evaluate the influence of the CTSA Program, as they work to anticipate what will be needed for continuing success. Themes include (1) considering the needs of stakeholders, (2) the perspective of the evaluators, (3) the importance of service improvement, (4) the importance of teams and people, (5) costs and return on investments, (6) methodology considerations to evaluate the CTSA enterprise, (7) innovation in evaluation, (8) defining the transformation of research, (9) evaluating the long-term impact of the CTSAs on public health, and (10) contributing to science policy formulation and implementation. The establishment of the CTSA Program, with its mandated evaluation component, has not only influenced the infrastructure and nature of translational research but will continue to impact policy and management in science.


Boosting the free fatty acid synthesis of Escherichia coli by expression of a cytosolic Acinetobacter baylyi thioesterase.
Thioesterases remove the fatty acyl moiety from the fatty acyl-acyl carrier proteins (ACPs), releasing them as free fatty acids (FFAs), which can be further used to produce a variety of fatty acid-based biofuels, such as biodiesel, fatty alcohols and alkanes. Thioesterases play a key role in the regulation of the fatty acid synthesis in Escherichia coli. Therefore, exploring more promising thioesterases will contribute to the development of industrial microbial lipids production. We cloned and expressed a cytosolic Acinetobacter baylyi thioesterase ('AcTesA) in E. coli by deleting its leader sequence. Protein sequence alignment, structure modeling and site-directed mutagenesis demonstrated that Ser10, Gly48, Asn77, Asp158 and His161 residues composed the active centre of 'AcTesA. The engineered strain that overexpressed 'AcTesA achieved a FFAs titer of up to 501.2 mg/L in shake flask, in contrast to only 20.5 mg/L obtained in wild-type E. coli, demonstrating that the expression of 'AcTesA indeed boosted the synthesis of FFAs. The 'AcTesA exhibited a substrate preference towards the C8-C16 acyl groups, with C14:0, C16:1, C12:0 and C8:0 FFAs being the top four components. Optimization of expression level of 'AcTesA made the FFAs production increase to 551.3 mg/L. The FFAs production further increased to 716.1 mg/L by optimization of the culture medium. Fed-batch fermentation was also carried out to evaluate the FFAs production in a scaleable process. Finally, 3.6 g/L FFAs were accumulated within 48 h, and a maximal FFAs yield of 6.1% was achieved in 12-16 h post induction. For the first time, an A. baylyi thioesterase was cloned and solubly expressed in the cytosol of E. coli. This leaderless thioesterase ('AcTesA) was found to be capable of enhancing the FFAs production of E. coli. Without detailed optimization of the strain and fermentation, the finally achieved 3.6 g/L FFAs is encouraging. In addition, 'AcTesA exhibited different substrate specificity from other thioesterases previously reported, and can be used to supply the fatty acid-based biofuels with high quality of FFAs. Altogether, this study provides a promising thioesterase for FFAs production, and is of great importance in enriching the library of useful thioesterases.


Clinical characteristics of 26 human cases of highly pathogenic avian influenza A (H5N1) virus infection in China.
While human cases of highly pathogenic avian influenza A (H5N1) virus infection continue to increase globally, available clinical data on H5N1 cases are limited. We conducted a retrospective study of 26 confirmed human H5N1 cases identified through surveillance in China from October 2005 through April 2008. Data were collected from hospital medical records of H5N1 cases and analyzed. The median age was 29 years (range 6-62) and 58% were female. Many H5N1 cases reported fever (92%) and cough (58%) at illness onset, and had lower respiratory findings of tachypnea and dyspnea at admission. All cases progressed rapidly to bilateral pneumonia. Clinical complications included acute respiratory distress syndrome (ARDS, 81%), cardiac failure (50%), elevated aminotransaminases (43%), and renal dysfunction (17%). Fatal cases had a lower median nadir platelet count (64.5 x 10(9) cells/L vs 93.0 x 10(9) cells/L, p = 0.02), higher median peak lactic dehydrogenase (LDH) level (1982.5 U/L vs 1230.0 U/L, p = 0.001), higher percentage of ARDS (94% [n = 16] vs 56% [n = 5], p = 0.034) and more frequent cardiac failure (71% [n = 12] vs 11% [n = 1], p = 0.011) than nonfatal cases. A higher proportion of patients who received antiviral drugs survived compared to untreated (67% [8/12] vs 7% [1/14], p = 0.003). The clinical course of Chinese H5N1 cases is characterized by fever and cough initially, with rapid progression to lower respiratory disease. Decreased platelet count, elevated LDH level, ARDS and cardiac failure were associated with fatal outcomes. Clinical management of H5N1 cases should be standardized in China to include early antiviral treatment for suspected H5N1 cases.


Prevalence of inflammatory back pain in a cohort of patients with anterior uveitis.
To determine the prevalence of inflammatory back pain in an anterior uveitis cohort. Retrospective cohort study. Patients with anterior uveitis were recruited from the clinic of an ophthalmologist to complete a survey between March and December 2008. Patients were classified with inflammatory back pain if they had ≥2 positive responses to 4 validated inflammatory back pain questions: presence of morning stiffness >30 minutes in duration; improvement in back pain with exercise but not with rest; awakening from back pain during the second half of the night only; and presence of alternating buttock pain. Disease activity was assessed using the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). The impact of disease on quality of life was measured using the EuroQOL (EQ-5D) questionnaire. Twenty-five patients underwent further rheumatologic examination. One hundred forty-one of 167 patients (84.4%) completed the survey. Sixty-six of 141 patients (46.8%) were classified to have inflammatory back pain. Mean BASDAI (4.2, SD 2.41) and EQ-5D scores (0.73, SD 0.21) were lower than patients with no inflammatory back pain (0.82, SD 0.16, P = .0048). In the subgroup that underwent rheumatologic assessment, a classification of inflammatory back pain was 92% sensitive and 67% specific for a diagnosis of inflammatory back pain. The prevalence of inflammatory back pain in a cohort of anterior uveitis patients was found to be 46.8%. Patients with inflammatory back pain had worse quality of life than those without. Ophthalmologists may use these questions on back pain to select patients classified to have inflammatory back pain to refer for early rheumatologic assessment.


Genetic investigations in childhood deafness.
Permanent childhood sensorineural hearing loss, is one of the most common birth defects in developed countries. It is important to identify the aetiology of hearing loss for many reasons, as there may be important health surveillance implications particularly with syndromic causes. Non-syndromic sensorineural hearing loss is a highly heterogeneous genetic condition, meaning that it may be caused by any one of numerous genes, with very few phenotypic distinctions between the different genetic types. This has previously presented significant challenges for genetic testing. However, the introduction of new technologies should enable more comprehensive testing in the future, bringing significant benefits to more affected children and their families.


Rab1A over-expression prevents Golgi apparatus fragmentation and partially corrects motor deficits in an alpha-synuclein based rat model of Parkinson's disease.
Although the overabundance of human alpha-synuclein in nigral dopaminergic neurons is considered to play a pathogenic role in Parkinson's disease (PD), it remains unclear how alpha-synuclein leads to neuronal degeneration and motor symptoms. Here, we explored the effect of human alpha-synuclein in the rat substantia nigra following AAV-mediated gene delivery inducing a moderate loss of dopaminergic neurons together with motor impairments. A significant fraction of the surviving nigral neurons were found to express human αSyn and displayed a pathological fragmentation of the Golgi apparatus. This observation prompted further investigation on the role of the secretory pathway, in particular at the ER/Golgi level, in alpha-synuclein toxicity. To address this question, we co-expressed human alpha-synuclein with Rab1A, a regulator of ER-to-Golgi vesicular trafficking, and found a significant reduction of Golgi fragmentation. Rab1A did not protect the dopaminergic neurons from the alpha-synuclein-induced degeneration that occurred within several months following vector injection. However, we observed in animals co-expressing Rab1A an improvement of motor behavior that correlates with the rescue of normal Golgi morphology in alpha-synuclein-expressing dopaminergic neurons. The non-prenylable mutant Rab1A-DeltaCC did not produce any of the effects observed with the wild-type form of Rab1A, linking the protective role of Rab1A with its activity in ER-to-Golgi vesicular trafficking. In conclusion, Rab1A can rescue the Golgi fragmentation caused by the overabundance of alpha-synuclein in nigral dopaminergic neurons, improving the ability of the surviving neurons to control motor function in hemiparkinsonian animals.


Excited state proton transfer based fluorescent molecular probes and their application in studying lipid bilayer membranes.
Molecules with ionizable protons, with different proton transfer efficiencies in the excited and ground states, show excited state prototropism (ESPT). In suitable proton donating/accepting environments, ESPT of a molecule can result in the observation of emissions from different prototropic species, each characterized by different emission spectra and different emission lifetimes. In condensed media, the immediate environment around the ESPT molecule can significantly influence the emission spectral parameters of different prototropic species. This forms the basis of ESPT based fluorescence sensing. The concept of ESPT has been widely used for probing dynamical and structural information of micro-heterogeneous media like micelles, polymers, lipid bilayer membranes, etc. ESPT molecules like naphthol and intra-molecular ESPT (ESIPT) molecules like hydroxyflavones etc., are said to be good multi-state fluorescent molecular probes if (i) the partitioning of these molecules to a micro-heterogeneous organized medium is more efficient, and (ii) the molecules possess distinct excitation and emission wavelengths corresponding to their different prototropic forms. The fluorescence of different prototropic forms shows a sensitive response towards the change in the local environment around the micro-heterogeneous organized medium concerning the physical properties, local structure, and dynamics. This review mainly comprises the work carried out on ESPT fluorescence molecular probing of biomimetic liposomes/lipid bilayer membranes from 1990 onwards.


The Progression of the Stargardt Disease Type 4 (ProgStar-4) Study: Design and Baseline Characteristics (ProgStar-4 Report No. 1).
To describe the design and baseline characteristics of patients enrolled in the multicenter, prospective natural history study of Stargardt disease type 4. Fifteen eligible patients aged 6 years and older at baseline, harboring disease-causing variants in the PROM1 gene, and with specified ocular lesions were enrolled. They were examined at baseline using a standard protocol, with 6 monthly follow-up visits for a 2-year period including best-corrected ETDRS visual acuity, spectral-domain optical coherence tomography, fundus autofluorescence (FAF), mesopic and scotopic microperimetry (MP). Areas of definitely decreased FAF (DDAF) and questionably decreased FAF were outlined and quantified on FAF images. Amongst the 15 patients (29 eyes) that were enrolled at 5 centers in the USA and Europe, 10 eyes (34.5%) had areas of DDAF with an average lesion area of 3.2 ± 3.5 mm2 (range 0.36-10.39 mm2) at baseline. The mean retinal sensitivity of the posterior pole derived from mesopic MP was 8.8 ± 5.8 dB. Data on disease progression in PROM1-related retinopathy from this study will contribute to the characterization of the natural history of disease and the exploration of the utility of several modalities to track progression and therefore to potentially be used in future interventional clinical trials.


Alcohol use during the great recession of 2008-2009.
The aim of this study was to assess changes in alcohol use in the USA during the Great Recession. Drinking participation, drinking frequency, drinking intensity, total alcohol consumption and frequency of binge drinking were assessed in a nationally representative sample of 2,050,431 US women and men aged 18 and older, interviewed between 2006 and 2010. The prevalence of any alcohol use significantly declined during the economic recession, from 52.0% in 2006-2007 to 51.6% in 2008-2009 (P < 0.05), corresponding to 880,000 fewer drinkers (95% confidence interval [CI] 140,000 to 1.6 million). There was an increase, however, in the prevalence of frequent binging, from 4.8% in 2006-2007 to 5.1% in 2008-2009 (P < 0.01), corresponding to 770,000 more frequent bingers (95% CI 390,000 to 1.1 million). Non-Black, unmarried men under 30 years, who recently became unemployed, were at highest risk for frequent binging. During the Great Recession there was an increase in abstention from alcohol and a rise in frequent binging.


Arguments in favour of compulsory treatment of opioid dependence.
Twelve agencies of the United Nations, including the World Health Organization, have issued a joint statement that calls on Member States to replace the compulsory detention of people who use opioids in treatment centres with voluntary, evidence-informed and rights-based health and social services. The arguments in favour of this position fall into three broad categories: Compulsory treatment centres infringe on an individual's liberty, they put human beings at risk of harm, and evidence of their effectiveness against opioid dependence has not been generated. The United Nations statement underscores that although countries apply different criteria for sending individuals to compulsory treatment centres, detention often takes place without due process, legal safeguards or judicial review. This clearly violates internationally recognized human rights standards. Furthermore, people who are committed to these centres are often exposed to physical and sexual violence, forced labour and sub-standard living conditions. They are often denied health care, despite their heightened vulnerability to HIV infection and tuberculosis. Finally, there is no evidence, according to the statement, that these centres offer an environment that is conducive to recovery from opioid dependence or to the rehabilitation of commercial sex workers or of children who have suffered sexual exploitation, abuse or lack of care and protection. The author of this paper sets forth several arguments that counter the position taken by the United Nations and argues in favour of compulsory treatment within a broader harm reduction strategy aimed at protecting society as well as the individual concerned.


Risk Denial and Socio-Economic Factors Related to High HIV Transmission in a Fishing Community in Rakai, Uganda: A Qualitative Study.
In Kasensero fishing community, home of the first recorded case of HIV in Uganda, HIV transmission is still very high with an incidence of 4.3 and 3.1 per 100 person-years in women and men, respectively, and an HIV prevalence of 44%, reaching up to 74% among female sex workers. We explored drivers for the high HIV transmission at Kasensero from the perspective of fishermen and other community members to inform future policy and preventive interventions. 20 in-depth interviews including both HIV positive and HIV negative respondents, and 12 focus-group discussions involving a total of 92 respondents from the Kasensero fishing community were conducted during April-September 2014. Content analysis was performed to identify recurrent themes. The socio-economic risk factors for high HIV transmission in Kasensero fishing community cited were multiple and cross-cutting and categorized into the following themes: power of money, risk denial, environmental triggers and a predisposing lifestyle and alcoholism and drug abuse. Others were: peer pressure, poor housing and the search for financial support for both the men and women which made them vulnerable to HIV exposure and or risk behavior. There is a need for context specific combination prevention interventions in Kasensero that includes the fisher folk and other influential community leaders. Such groups could be empowered with the knowledge and social mobilization skills to fight the negative and risky behaviors, perceptions, beliefs, misconceptions and submission attitudes to fate that exposes the community to high HIV transmission. There is also need for government/partners to ensure effective policy implementation, life jackets for all fishermen, improve the poor housing at the community so as to reduce overcrowding and other housing related predispositions to high HIV rates at the community. Work place AIDS-competence teams have been successfully used to address high HIV transmission in similar settings.


Absence of mannose-binding lectin prevents hyperglycemic cardiovascular complications.
Diabetes, stress, pharmaceuticals, surgery, and physical trauma can lead to hyperglycemic conditions. A consistent relationship has been found between chronic inflammation and the cardiovascular complications of hyperglycemia. We hypothesized that cardiomyopathy and vasculopathy resulting from acute hyperglycemia are dependent on mannose-binding lectin (MBL) and lectin complement pathway activation. Hyperglycemia was induced in wild-type (WT) C57BL/6 and MBL-null mice after streptozotocin administration. Echocardiographic data and tissue samples were collected after 4, 7, or 14 days of acute hyperglycemia. Hyperglycemic WT mice demonstrated dilated cardiomyopathy with significantly increased short and long axis area measurements during systole and diastole compared to hyperglycemic MBL-null mice. The EC(50) for acetylcholine-induced relaxation of mesenteric arterioles in WT mice after 4 days of hyperglycemia demonstrated a significant loss of nitric oxide-mediated relaxation compared to normoglycemic WT or hyperglycemic MBL-null mice. Myocardial histochemistry and Western blot analysis revealed a significant influx of macrophages, altered morphology, and increased elastin and collagen deposition in hyperglycemic WT hearts compared to MBL-null hearts. Serum transforming growth factor-β1 levels were significantly lower in hyperglycemic MBL-null compared to WT mice, suggesting decreased profibrotic signaling. Together, these data suggest that MBL and the lectin complement pathway play a significant role in vascular dysfunction and cardiomyopathy after acute hyperglycemia.


Admission ECG predicts long-term outcome in acute coronary syndromes without ST elevation.
Acute coronary syndromes (ACS) without ST elevation are a frequent cause of hospital admission, myocardial infarction and death. To explore the role of the ECG in stratifying ACS patients. Prospective, centrally-coordinated multicentre registry involving 56 centres throughout the UK. Consecutive patients admitted with ACS without ST elevation on the presenting ECG (n = 1046) were followed for 6 months. A subgroup (n = 653) were flagged with the UK Office for National Statistics and followed-up for death over 4 years. Mean follow-up for the group as a whole was 2.4 years. In the first 6 months, the death rate was 7.3%. Survival at 1 year was 90.8% (95%CI 88.2%-92.8%); at 45 months it was 77.8% (95%CI 74.1%-81.1%). We compared data in those with ST depression or bundle branch block on the admission ECG (n = 304, 29%) with those with T wave inversion, Q waves and minor ST segment changes (n = 576, 55%) and those with a normal ECG (n = 166, 16%). Their respective incidences of death were 15%, 5% and 2% (p < 0.01) at 6 months, and 38%, 22% and 7% (p < 0.01) at 4 years. Rates of adverse events are high in patients admitted to UK hospitals with ACS without ST elevation. The ECG remains a very important and simple discriminator of both short- and long-term risk, enabling more aggressive, proven therapies to be targeted towards those at highest risk.


Comparative effectiveness of injectable paliperidone palmitate versus oral atypical antipsychotics: early postmarketing evidence.
To provide comparative effectiveness evidence for long-acting injectable paliperidone palmitate versus oral atypical antipsychotics. We performed a retrospective, observational cohort study using patient claims data from Missouri Medicaid to compare the likelihood of emergency department (ED) visits and hospitalizations in the year following drug initiation using multivariable logistic regression. Adjusted odds ratios (AOR) for ED visits (AOR: 0.63; 95% CI: 0.47-0.85) and hospitalizations (AOR: 0.85; 95% CI: 0.64-1.13) were lower in paliperidone palmitate patients, although hospitalizations did not achieve statistical significance. Sensitivity analyses examining mental health-related outcomes and using different analytic strategies for patient selection bias showed directionally similar beneficial effects but were not statistically significant. Early evidence for paliperidone palmitate under real-world conditions is encouraging. However, caution should be taken until additional research substantiates the findings with greater certainty.


Microbiological characteristics of acute osteoarticular infections in children.
This study aimed to describe the microbiological characteristics of acute septic arthritis (SA) and osteomyelitis (OM) in children. Cases of children (0-15 years) with SA/OM were identified through a retrospective search of hospital discharge codes over a six-year period. In addition, a systematic literature search and meta-analysis of studies reporting culture results of children with SA/OM was performed. In our retrospective chart review, we identified 65 cases of OM and 46 cases of SA. The most frequently cultured organisms in both conditions were Gram-positive cocci, primarily Staphylococcus aureus. On admission, most patients had a normal white blood cell count (WCC) but elevated C-reactive protein (CRP) and/or erythrocyte sedimentation rate (ESR). Bacteraemia was associated with a longer mean length of hospitalization for both infections. Considering our results and the meta-analysis, we found low rates of culture-positivity in cases of clinically confirmed infection. In SA, articular fluid was culture-positive in 42.49% [95% confidence interval (CI) 28.39-57.23]. In OM, intra-operative samples were culture-positive in 52.65% (95% CI 30.54-74.22). Bacteraemia was detected in 23.91% (95% CI 8.40-44.24) of children with SA and 21.48% (95% CI 10.89-34.47) with OM. Despite appropriate sampling, a positive microbiological diagnosis is often lacking in paediatric acute osteoarticular infection using standard culture-based methods. This highlights the need for validation and use of more sensitive diagnostic methods, such as PCR.


High Contrast Allows the Retina to Compute More Than Just Contrast.
The goal of sensory processing is to represent the environment of an animal. All sensory systems share a similar constraint: they need to encode a wide range of stimulus magnitudes within their narrow neuronal response range. The most efficient way, exploited by even the simplest nervous systems, is to encode relative changes in stimulus magnitude rather than the absolute magnitudes. For instance, the retina encodes contrast, which are the variations of light intensity occurring in time and in space. From this perspective, it is easy to understand why the bright plumage of a moving bird gains a lot of attention, while an octopus remains motionless and mimics its surroundings for concealment. Stronger contrasts simply cause stronger visual signals. However, the gains in retinal performance associated with higher contrast are far more than what can be attributed to just a trivial linear increase in signal strength. Here we discuss how this improvement in performance is reflected throughout different parts of the neural circuitry, within its neural code and how high contrast activates many non-linear mechanisms to unlock several sophisticated retinal computations that are virtually impossible in low contrast conditions.


Treatment actions and treatment failure: case studies in the response to severe childhood febrile illness in Mali.
Appropriate home management of illness is vital to efforts to control malaria. The strategy of home management relies on caregivers to recognize malaria symptoms, assess severity and promptly seek appropriate care at a health facility if necessary. This paper examines the management of severe febrile illness (presumed malaria) among children under the age of five in rural Koulikoro Region, Mali. This research examines in-depth case studies of twenty-five households in which a child recently experienced a severe febrile illness, as well as key informant interviews and focus group discussions with community members. These techniques were used to explore the sequence of treatment steps taken during a severe illness episode and the context in which decisions were made pertaining to pursing treatments and sources of care, while incorporating the perspective and input of the mother as well as the larger household. Eighty-one participants were recruited in 25 households meeting inclusion criteria. Children's illness episodes involved multiple treatment steps, with an average of 4.4 treatment steps per episode (range: 2-10). For 76% of children, treatment began in the home, but 80% were treated outside the home as a second recourse. Most families used both traditional and modern treatments, administered either inside the home by family members, or by traditional or modern healers. Participants' stated preference was for modern care, despite high rates of reported treatment failure (52%, n=12), however, traditional treatments were also often deemed appropriate and effective. The most commonly cited barrier to seeking care at health facilities was cost, especially during the rainy season. Financial constraints often led families to use traditional treatments. Households have few options available to them in moments of overlapping health and economic crises. Public health research and policy should focus on the reducing barriers that inhibit poor households from promptly seeking appropriate health care. Enhancing the quality of care provided at community health facilities and supporting mechanisms by which treatment failures are quickly identified and addressed can contribute to reducing subsequent treatment delays and avoid inappropriate recourse to traditional treatments.


The Effect of Articular Cartilage Focal Defect Size and Location in Whole Knee Biomechanics Models.
Articular cartilage focal defects are common soft tissue injuries potentially linked to osteoarthritis (OA) development. Although several defect characteristics likely contribute to osteoarthritis, their relationship to local tissue deformation remains unclear. Using finite element models with various femoral cartilage geometries, we explore how defects change cartilage deformation and joint kinematics assuming loading representative of the maximum joint compression during the stance phase of gait. We show how defects, in combination with location-dependent cartilage mechanics, alter deformation in affected and opposing cartilages, as well as joint kinematics. Small and average sized defects increased maximum compressive strains by approximately 50% and 100%, respectively, compared to healthy cartilage. Shifts in the spatial locations of maximum compressive strains of defect containing models were also observed, resulting in loading of cartilage regions with reduced initial stiffnesses supporting the new, elevated loading environments. Simulated osteoarthritis (modeled as a global reduction in mean cartilage stiffness) did not significantly alter joint kinematics, but exacerbated tissue deformation. Femoral defects were also found to affect healthy tibial cartilage deformations. Lateral femoral defects increased tibial cartilage maximum compressive strains by 25%, while small and average sized medial defects exhibited decreases of 6% and 15%, respectively, compared to healthy cartilage. Femoral defects also affected the spatial distributions of deformation across the articular surfaces. These deviations are especially meaningful in the context of cartilage with location-dependent mechanics, leading to increases in peak contact stresses supported by the cartilage of between 11% and 34% over healthy cartilage.


Inhibition of DAT function attenuates manganese accumulation in the globus pallidus.
Manganese (Mn) is an essential nutrient, though exposure to high concentrations may result in neurotoxicity characterized by alterations in dopamine neurobiology. To date, it remains elusive how and why Mn targets dopaminergic neurons although recently the role of the dopamine transporter has been suggested. Our primary goal of this study was to examine the potential roles of the monoamine transporters, dopamine transporter (DAT), serotonin transporter (SERT) and norepinephrine transporter (NET), in neuronal Mn transport. Using striatal synaptosomes, we found that only inhibition of DAT significantly decreased Mn accumulation. Furthermore, weanling rats chronically exposed to Mn, significantly accumulated Mn in several brain regions. However, rats receiving the specific DAT inhibitor GBR12909 (1 mg/kg bw, three times/week; four weeks) had significantly lower Mn levels only in the globus pallidus compared to saline-treated rats (p<0.05). Our data show that inhibition of DAT exclusively inhibits Mn accumulation in the globus pallidus during chronic exposure.


Anal sex is a behavioural marker for laboratory-confirmed vaginal sexually transmissible infections and HIV-associated risk among African-American female adolescents.
African-American females are disproportionately affected by HIV and sexually transmissible infections (STIs). The prevalence of anal sex and its association with other sexual risk behaviours is understudied in this population. Participants were 715 African-American females, 15 to 21 years old, who had reported sexual activity in the previous 60 days. Data collection included an audiocomputer assisted self-interview (ACASI) and a self-collected vaginal swab specimen assayed using nucleic acid amplification tests to detect the presence of Chlamydia trachomatis and Neisseria gonorrhoeae, and real-time polymerase chain reaction assay to detect Trichomonas vaginalis. Approximately 10.5% reported anal sex, at least once, during the 60 days before completing the computerised baseline assessment. The prevalence of any STI was significantly greater among adolescents reporting recent anal sex (40% tested positive for at least one of three laboratory-confirmed STIs) relative to those adolescents not reporting anal sex (27.5% STI prevalence). Of the 10 outcomes comprising the sexual risk profile, seven achieved bivariate significance, with each of the differences indicating greater risk for those recently engaging in anal sex. In multivariable controlled analyses, six of the seven measures retained statistical significance. African-American adolescent females who engage in penile-anal sex may experience an elevated risk of vaginally-acquired STIs. The findings suggest that, among those having penile-anal sex, several HIV/STI-associated sexual risk behaviours are significantly more prevalent. Thus, penile-anal sex may be an important proxy of overall sexual risk behaviours and can be readily assessed during paediatrician visits as part of a sexual history.


Synergistic effect of hyaluronate fragments in retinaldehyde-induced skin hyperplasia which is a Cd44-dependent phenomenon.
CD44 is a polymorphic proteoglycan and functions as the principal cell-surface receptor for hyaluronate (HA). Heparin-binding epidermal growth factor (HB-EGF) activation of keratinocyte erbB receptors has been proposed to mediate retinoid-induced epidermal hyperplasia. We have recently shown that intermediate size HA fragments (HAFi) reverse skin atrophy by a CD44-dependent mechanism. Treatment of primary mouse keratinocyte cultures with retinaldehyde (RAL) resulted in the most significant increase in keratinocyte proliferation when compared with other retinoids, retinoic acid, retinol or retinoyl palmitate. RAL and HAFi showed a more significant increase in keratinocyte proliferation than RAL or HAFi alone. No proliferation with RAL was observed in CD44-/- keratinocytes. HA synthesis inhibitor, 4-methylumbelliferone inhibited the proliferative effect of RAL. HB-EGF, erbB1, and tissue inhibitor of MMP-3 blocking antibodies abrogated the RAL- or RAL- and HAFi-induced keratinocyte proliferation. Topical application of RAL or RAL and HAFi for 3 days caused a significant epidermal hyperplasia in the back skin of wild-type mice but not in CD44-/- mice. Topical RAL and HAFi increased epidermal CD44 expression, and the epidermal and dermal HA. RAL induced the expression of active HB-EGF and erbB1. However, treatment with RAL and HAFi showed a more significant increase in pro-HB-EGF when compared to RAL or HAFi treatments alone. We then topically applied RAL and HAFi twice a day to the forearm skin of elderly dermatoporosis patients. After 1 month of treatment, we observed a significant clinical improvement. Our results indicate that (i) RAL-induced in vitro and in vivo keratinocyte proliferation is a CD44-dependent phenomenon and requires the presence of HA, HB-EGF, erbB1 and MMPs, (ii) RAL and HAFi show a synergy in vitro and in vivo in mouse skin, and (iii) the combination of RAL and HAFi seems to have an important therapeutic effect in dermatoporosis.


Assessing the protein concentration in commercial enzyme preparations.
Although a poor indicator of how a cellulase preparation will perform on biomass, the filter paper unit (FPU) still finds wide use in the literature as an apparent measure of performance efficacy. In actuality, the assessment of commercial enzyme preparation performance in terms of biomass conversion or solubilization of insoluble polysaccharides is largely dependent on the substrate composition, which cannot be easily standardized. Commercial cellulase preparations are evaluated based upon their performance or specific activity. The ability to compare commercial enzyme preparation efficacy across a wide variety of different preparations requires defining the amount of enzyme protein required in milligrams per gram of cellulose to achieve a targeted level of cellulose hydrolysis in a specified timeframe. Since biomass substrates are highly variable, reproducible and accurate protein determination is as important as performance testing to be able to rank order the effectiveness of diverse preparations. This chapter describes a protocol that overcomes many of the difficulties encountered with determining the protein concentration in commercial cellulase preparations.


Sniffing out olfactory reference syndrome.
Olfactory reference syndrome is characterised by the erroneous belief that one emits an unpleasant body odour. This results in significant distress and is often accompanied by repetitive behaviour such as frequent showering in an attempt to camouflage the perceived odour. The body odour concerns may have a delusional quality and do not respond to simple reassurance or counterexample. Herein, we report the case of an olfactory reference disorder (ORD) patient who had received multiple medical interventions and undergone polysurgery prior to an accurate diagnosis being established. ORD may lead to significant disability, yet often goes unrecognised for many years. For many patients, poor insight will contribute to their reluctance to consider psychiatric treatment. This case demonstrated that a multimodal treatment approach comprising judicious medication use, combined with cognitive behavioural therapy, in the context of a therapeutic alliance yielded therapeutic success.


Radiological Patterns of Brain Metastases in Breast Cancer Patients: A Subproject of the German Brain Metastases in Breast Cancer (BMBC) Registry.
Evidence about distribution patterns of brain metastases with regard to breast cancer subtypes and its influence on the prognosis of patients is insufficient. Clinical data, cranial computed tomography (CT) and magnetic resonance imaging (MRI) scans of 300 breast cancer patients with brain metastases (BMs) were collected retrospectively in four centers participating in the Brain Metastases in Breast Cancer Registry (BMBC) in Germany. Patients with positive estrogen (ER), progesterone (PR), or human epidermal growth factor receptor 2 (HER2) statuses, had a significantly lower number of BMs at diagnosis. Concerning the treatment mode, HER2-positive patients treated with trastuzumab before the diagnosis of BMs showed a lower number of intracranial metastases (p < 0.001). Patients with a HER2-positive tumor-subtype developed cerebellar metastases more often compared with HER2-negative patients (59.8% vs. 44.5%, p = 0.021), whereas patients with triple-negative primary tumors had leptomeningeal disease more often (31.4% vs. 18.3%, p = 0.038). The localization of Brain metastases (BMs) was associated with prognosis: patients with leptomeningeal disease had shorter survival compared with patients without signs of leptomeningeal disease (median survival 3 vs. 5 months, p = 0.025). A shorter survival could also be observed in the patients with metastases in the occipital lobe (median survival 3 vs. 5 months, p = 0.012). Our findings suggest a different tumor cell homing to different brain regions depending on subtype and treatment.


Sensory Guillain-Barré syndrome and related disorders: an attempt at systematization.
The possibility that some patients diagnosed with an acute sensory neuropathy could actually have Guillain-Barré syndrome (GBS) has been repeatedly advanced in the literature, but the number of cases reported is small. The reports have shown different clinical presentations and electrophysiological findings and are variously named, thus generating terminological and nosological confusion. We operatively defined sensory GBS as an acute, monophasic, widespread neuropathy characterized clinically by exclusive sensory symptoms and signs that reach their nadir in a maximum of 6 weeks without related systemic disorders and other diseases or conditions. We reviewed the literature through searches of PubMed from 1980 to March 2011 and our own files. On the basis of the size of fibers involved and the possible site of primary damage, we propose tentatively classifying sensory GBS and related disorders into three subtypes: acute sensory demyelinating polyneuropathy; acute sensory large-fiber axonopathy-ganglionopathy; and acute sensory small-fiber neuropathy-ganglionopathy.


The role of conformational heterogeneity in regulating the apoptotic activity of BAX protein.
While activation of BAX is required for initiating mitochondria-mediated apoptosis, the underlying mechanisms remain unsettled. We studied conformations of BAX protein using pressure- and temperature-resolved ESR techniques and obtained the thermodynamic properties of the conformations. We show that inactive BAX is structurally heterogeneous and exists in equilibrium between two major populations of the conformations, UM and UM', of which the former is thermodynamically favored at room temperature. An increase in the population of UM', induced by either pressure or point mutations of BAX, renders BAX susceptible to oligomerization, which leads to cell death. This study uncovers the biological significance of BAX conformations and shows that the pro-apoptotic activity of BAX can be triggered by altering the equilibrium between the two states. It suggests that therapeutic intervention may focus on shifting the balance in the conformational heterogeneity.


Measurement characteristics of two asthma symptom diary scales for use in clinical trials.
The measurement characteristics of two asthma symptom diary scales developed for use as health outcome measures in clinical trials of asthma therapy were investigated. A daytime diary scale was designed to capture the frequency and inconvenience of daytime asthma symptoms and their effects on activities, and a nocturnal asthma symptom diary scale was designed to capture awakenings with asthma symptoms. The internal consistency, reliability, validity and responsiveness of both asthma diary scales were assessed in 346 adult asthma patients in two placebo-controlled clinical trials of an investigational asthma therapy, a leukotriene biosynthesis inhibitor. The daytime symptom scale showed sufficient internal consistency (0.90-0.92), and the daytime and nocturnal symptom scales showed sufficient test retest reliability (0.69-0.87). Construct validity was demonstrated by generally moderate-to-strong correlations for changes in the diary scales with changes in other measures of asthma status, such as forced expiratory volume in one second (FEV1), peak expiratory flow (PEF), and puffs of beta-agonist inhaler. Both scales demonstrated significant responsiveness to change in asthma due to therapy in one of the clinical trials. Based on these results, the daytime and nocturnal asthma symptom diary scales show measurement characteristics appropriate for use as asthma outcome measures in clinical trials of asthma therapy.


Analysis of misalignment-induced measurement error for goniophotometry of light-emitting diode arrays.
The luminous distribution characteristics of light sources can be measured by goniophotometry. In this work, the misalignment of luminary-induced measurement errors as the main factor affecting the measurement accuracy is analyzed. A calculation method for measurement error is proposed. Then, the translational and angular misalignment-induced measurement errors are calculated and analyzed. Results show that the measurement errors induced by misalignments may be great in some cases even if the far-field condition is satisfied. For luminaries with different radiation patterns, the acceptable misalignment tolerances corresponding to measurement error of less than 1% are given.


A global analysis of the impacts of urbanization on bird and plant diversity reveals key anthropogenic drivers.
Urbanization contributes to the loss of the world's biodiversity and the homogenization of its biota. However, comparative studies of urban biodiversity leading to robust generalities of the status and drivers of biodiversity in cities at the global scale are lacking. Here, we compiled the largest global dataset to date of two diverse taxa in cities: birds (54 cities) and plants (110 cities). We found that the majority of urban bird and plant species are native in the world's cities. Few plants and birds are cosmopolitan, the most common being Columba livia and Poa annua. The density of bird and plant species (the number of species per km(2)) has declined substantially: only 8% of native bird and 25% of native plant species are currently present compared with estimates of non-urban density of species. The current density of species in cities and the loss in density of species was best explained by anthropogenic features (landcover, city age) rather than by non-anthropogenic factors (geography, climate, topography). As urbanization continues to expand, efforts directed towards the conservation of intact vegetation within urban landscapes could support higher concentrations of both bird and plant species. Despite declines in the density of species, cities still retain endemic native species, thus providing opportunities for regional and global biodiversity conservation, restoration and education.


Kinesin-1 and mitochondrial motility control by discrimination of structurally equivalent but distinct subdomains in Ran-GTP-binding domains of Ran-binding protein 2.
The pleckstrin homology (PH) domain is a versatile fold that mediates a variety of protein-protein and protein-phosphatidylinositol lipid interactions. The Ran-binding protein 2 (RanBP2) contains four interspersed Ran GTPase-binding domains (RBD(n = 1-4)) with close structural homology to the PH domain of Bruton's tyrosine kinase. The RBD2, kinesin-binding domain (KBD) and RBD3 comprise a tripartite domain (R2KR3) of RanBP2 that causes the unfolding, microtubule binding and biphasic activation of kinesin-1, a crucial anterograde motor of mitochondrial motility. However, the interplay between Ran GTPase and R2KR3 of RanBP2 in kinesin-1 activation and mitochondrial motility is elusive. We use structure-function, biochemical, kinetic and cell-based assays with time-lapse live-cell microscopy of over 260,000 mitochondrial-motility-related events to find mutually exclusive subdomains in RBD2 and RBD3 towards Ran GTPase binding, kinesin-1 activation and mitochondrial motility regulation. The RBD2 and RBD3 exhibit Ran-GTP-independent, subdomain and stereochemical-dependent discrimination on the biphasic kinetics of kinesin-1 activation or regulation of mitochondrial motility. Further, KBD alone and R2KR3 stimulate and suppress, respectively, multiple biophysical parameters of mitochondrial motility. The regulation of the bidirectional transport of mitochondria by either KBD or R2KR3 is highly coordinated, because their kinetic effects are accompanied always by changes in mitochondrial motile events of either transport polarity. These studies uncover novel roles in Ran GTPase-independent subdomains of RBD2 and RBD3, and KBD of RanBP2, that confer antagonizing and multi-modal mechanisms of kinesin-1 activation and regulation of mitochondrial motility. These findings open new venues towards the pharmacological harnessing of cooperative and competitive mechanisms regulating kinesins, RanBP2 or mitochondrial motility in disparate human disorders.


The case-control study as data missing by design: estimating risk differences.
There are advantages to viewing the case-control design as a missing-data problem instead of as a sampling problem. In the simplest setup, cases are those members of a population who develop disease; controls can be a small random sample of the large number who do not; and covariates, including exposures and other important variables, are available only for cases and controls and are assumed to be missing at random for the remaining large fraction of the population. This approach allows estimation of the joint distribution of all variables in the population. Thus, when the size of the population is known, analysis is not restricted to logistic and other multiplicative intercept models. Methods based on this approach can obtain estimates and confidence intervals for parameters representing the effect of exposure on disease, with multivariate adjustment for other factors. Thus, case-control data can be used to estimate the risk difference, a parameter with great public health value. The missing-data perspective offers an additional advantage by linking the "study base principle" of control selection with the statistical concept of "missing at random." As an illustration, I use a subset of data from a case-control study to obtain estimates of the difference between annual risk of bladder cancer for various levels of smoking and lifetime non-smokers, adjusted for occupational exposure.


Ligand-field symmetry effects in Fe(II) polypyridyl compounds probed by transient X-ray absorption spectroscopy.
Ultrafast excited-state evolution in polypyridyl Fe(II) complexes is of fundamental interest for understanding the origins of the sub-ps spin-state changes that occur upon photoexcitation of this class of compounds as well as for the potential impact such ultrafast dynamics have on incorporation of these compounds in solar energy conversion schemes or switchable optical storage technologies. We have demonstrated that ground-state and, more importantly, ultrafast time-resolved X-ray absorption methods can offer unique insights into the interplay between electronic and geometric structure that underpins the photo-induced dynamics of this class of compounds. The present contribution examines in greater detail how the symmetry of the ligand field surrounding the metal ion can be probed using these X-ray techniques. In particular, we show that steady-state K-edge spectroscopy of the nearest-neighbour nitrogen atoms reveals the characteristic chemical environment of the respective ligands and suggests an interesting target for future charge-transfer femtosecond and attosecond spectroscopy in the X-ray water window.


Postmortem prolactin as a marker of antemortem stress.
To determine whether raised prolactin concentrations could be identified using postmortem venous blood and whether the level of prolactin correlated with antemortem stress. Blood was obtained from the right femoral vein of 100 random adult necropsy cases, all of whom had been dead less than four days. Prolactin was determined in the samples by microparticle immunoassay. The levels of prolactin obtained were then analysed in relation to sex and cause of death, with particular emphasis on a history of antemortem stress and drug use. Prolactin in all cases of trauma was in the normal range (up to 500 mU/l). In cases of sudden unexpected deaths the mean concentration was 533 mU/l (95% confidence interval (CI), 372 to 694 mU/l). Postoperative deaths or cases with chronic disease had a mean value of 1027 mU/l (95% CI, 735 to 1319 mU/l). Cases of suicide had a mean value of 1398 mU/l. Analysis of the suicides by sex showed a significant difference, the mean in female cases being 2072 mU/l compared with 692 mU/l in male cases. In three of the four female suicides with the highest prolactin, the hyperprolactinaemia might have been attributable to a drug effect, but one case still had unexplained hyperprolactinaemia. It is possible to detect prolactin reliably in postmortem venous blood samples. Prolactin values at necropsy differ according to the cause of death, with markedly higher values in postoperative deaths and in the chronically ill. Hyperprolactinaemia in cases of suicide is likely to result from the effects of the drugs used, but the levels were higher than previously reported.


Regulation of eukaryotic gene expression by transactivating proteins and cis acting DNA elements.
In this review we try to examine some of the recent developments in our understanding of the mechanisms that control gene expression in eukaryotes. We discuss the nature of the positive regulation exerted by adenovirus, herpes virus or papova immediate early proteins. These proteins which can activate homologous promoters can also stimulate transcription from cellular promoters present on transfected DNA. This property of the E1a gene product of Adenovirus e.g. may be related to its immortalizing function. Transcription of cellular genes can be stimulated by viral or cellular short DNA elements named enhancers. These elements acting in cis, can be placed 5' or 3' to the gene and function in both orientations. Some of them show a remarkable cell specificity in their action. Enhancers affect the chromatin structure by creating a local nuclease sensitive region that may serve as an entry site for RNA polymerase II or for factors involved in the process of transcription initiation.


Experimental canine leishmaniasis: clinical, parasitological and serological follow-up.
Canine leishmaniasis (CanL) caused by Leishmania infantum is transmitted by the bite of phlebotomine sand flies and affects millions of dogs in Europe, Asia, North Africa and South America. Canis familiaris is the major host for these parasites, and the main reservoir for human visceral infection. The development of effective molecules for therapy and immunoprophylaxis, would be an important tool in the control of this zoonosis. The aim of this study was to characterize an experimental CanL model in order to determine the best challenge model and which parameters are the most reliable to evaluate the efficacy of new drugs or vaccine candidates against L. infantum infection. The intravenous challenge with purified amastigotes used in this study allowed the development of infection in all animals inoculated (as confirmed by the detection of parasite in the different tissues and organs collected 6 months after inoculation). Molecular and serologic techniques were efficient methods for the follow-up. Lymph node and bone marrow aspirates were suitable clinical samples to detect the presence of Leishmania parasites. Despite ELISA was highly sensitive in detecting specific anti-Leishmania antibodies the use of two tests can improve the sensitivity and specificity of serological diagnosis.


Production and modification of nanofibrillated cellulose using various mechanical processes: a review.
Nanofibrillated cellulose from biomass has recently gained attention owing to their biodegradable nature, low density, high mechanical properties, economic value and renewability. Although they still suffer from two major drawbacks. The first challenge is the exploration of raw materials and its application in nanocomposites production. Second one is high energy consumption regarding the mechanical fibrillation. However, pretreatments before mechanical isolation can overcome this problem. Hydrophilic nature of nano-size cellulose fibers restricts good dispersion of these materials in hydrophobic polymers and therefore, leads to lower mechanical properties. Surface modification before or after mechanical defibrillation could be a solution for this problem. Additionally, drying affects the size of nanofibers and its properties which needs to study further. This review focuses on recent developments in pretreatments, nanofibrillated cellulose production and its application in nanopaper applications, coating additives, security papers, food packaging, and surface modifications and also for first time its drying.


Relating induced changes in EEG signals to orientation of visual stimuli using the ESI-256 machine.
The focus of this study is to investigate the relations that exist between changes in the orientation of simple visual stimuli displayed to a subject and the induced changes in brain activity recorded as EEG signals. These signals are recorded using the Electric Source Imaging with 256 electrodes (ESI-256). The 256-channel EEG signals of four subjects were measured monopolarly. Each subject was stimulated visually for approximately 7.5 minutes. The stimuli consisted of a series of 300 images depicting four basic orientations of a simple graphical element: a white bar on a black background, with each one of the four orientations (horizontal, vertical, +45 degrees and -45 degrees) shown a total of 75 times in a random order. The notion of missing information under certain orientations is not addressed at this juncture. The EEG signals produced by each subject were recorded in a continuous mode using a sampling rate of 1 kHz. Pre-processing of the raw EEG data obtained consisted of epoching, exclusion of faulty electrodes, and reduction of electro-oculogram (EOG) noise due to eye blinks. Topographical maps displaying brain activities and their individual electrode recordings are used as two different means for assessing these changes. It is important to note that the simplicity of the visual stimuli was considered in view of the massive data collected for interpretation. Our goal is to observe and determine new measures that would allow for the quantification and interpretation of such EEG brain activities. Such findings might prove useful for the later use of more complex stimuli and the potential development of size and orientation independent algorithms in image processing.


Subpopulation-proteomics in prokaryotic populations.
Clonal microbial cells do not behave in an identical manner and form subpopulations during cultivation. Besides varying micro-environmental conditions, cell inherent features like cell cycle dependent localization and concentration of regulatory proteins as well as epigenetic properties are well accepted mechanisms creating cell heterogeneity. Another suspected reason is molecular noise on the transcriptional and translational level. A promising tool to unravel reasons for cell heterogeneity is the combination of cell sorting and subpopulation proteomics. This review summarizes recent developments in prokaryotic single-cell analytics and provides a workflow for selection of single cells, low cell number mass spectrometry, and proteomics evaluation. This approach is useful for understanding the dependency of individual cell decisions on inherent protein profiles.