Structure and stereospecificity of the dehydratase domain from the terminal module of the rifamycin polyketide synthase.
RifDH10, the dehydratase domain from the terminal module of the rifamycin polyketide synthase, catalyzes the stereospecific syn dehydration of the model substrate (2S,3S)-2-methyl-3-hydroxypentanoyl-RifACP10, resulting in the exclusive formation of (E)-2-methyl-2-pentenoyl-RifACP10. RifDH10 does not dehydrate any of the other three diastereomeric, RifACP10-bound, diketide thioester substrates. On the other hand, when EryACP6, from the sixth module of the erythromycin polyketide synthase, is substituted for RifACP10, RifDH10 stereospecifically dehydrates only (2R,3R)-2-methyl-3-hydroxypentanoyl-EryACP6 to give exclusively (E)-2-methyl-2-pentenoyl-EryACP6, with no detectable dehydration of any of the other three diastereomeric, EryACP6-bound, diketides. An identical alteration in substrate diastereospecificity was observed for the corresponding N-acetylcysteamine or pantetheine thioester analogues, regardless of acyl chain length or substitution pattern. Incubation of (2RS)-2-methyl-3-ketopentanoyl-RifACP10 with the didomain reductase-dehydratase RifKR10-RifDH10 yielded (E)-2-methyl-2-pentenoyl-RifACP10, the expected product of syn dehydration of (2S,3S)-2-methyl-3-hydroxypentanoyl-RifACP10, while incubation with the corresponding EryACP6-bound substrate, (2RS)-2-methyl-3-ketopentanoyl-EryACP6, gave only the reduction product (2S,3S)-2-methyl-3-hydroxypentanoyl-EryACP6 with no detectable dehydration. These results establish the intrinsic syn dehydration stereochemistry and substrate diastereoselectivity of RifDH10 and highlight the critical role of the natural RifACP10 domain in chaperoning the proper recognition and processing of the natural ACP-bound undecaketide substrate. The 1.82 Å resolution structure of RifDH10 reveals the atomic-resolution details of the active site and allows modeling of the syn dehydration of the (2S,3S)-2-methyl-3-hydroxyacyl-RifACP10 substrate. These results suggest that generation of the characteristic cis double bond of the rifamycins occurs after formation of the full-length RifACP10-bound acyclic trans-unsaturated undecaketide intermediate, most likely during the subsequent macrolactamization catalyzed by the amide synthase RifF.


The interplay between cyclic AMP, MAPK, and NF-κB pathways in response to proinflammatory signals in microglia.
Cyclic AMP is an important intracellular regulator of microglial cell homeostasis and its negative perturbation through proinflammatory signaling results in microglial cell activation. Though cytokines, TNF-α and IL-1β, decrease intracellular cyclic AMP, the mechanism by which this occurs is poorly understood. The current study examined which signaling pathways are responsible for decreasing cyclic AMP in microglia following TNF-α stimulation and sought to identify the role cyclic AMP plays in regulating these pathways. In EOC2 microglia, TNF-α produced a dramatic reduction in cyclic AMP and increased cyclic AMP-dependent PDE activity that could be antagonized by Rolipram, myristoylated-PKI, PD98059, or JSH-23, implicating a role for PDE4, PKA, MEK, and NF-κB in this regulation. Following TNF-α there were significant increases in iNOS and COX-2 immunoreactivity, phosphorylated ERK1/2 and NF-κB-p65, IκB degradation, and NF-κB p65 nuclear translocation, which were reduced in the presence of high levels of cyclic AMP, indicating that reductions in cyclic AMP during cytokine stimulation are important for removing its inhibitory action on NF-κB activation and subsequent proinflammatory gene expression. Further elucidation of the signaling crosstalk involved in decreasing cyclic AMP in response to inflammatory signals may provide novel therapeutic targets for modulating microglial cell activation during neurological injury and disease.


Marine Resources Offer New Compounds and Strategies for the Treatment of Skin and Soft Tissue Infections.
Bioprospecting of the marine environment for drug development has gained much attention in recent years owing to its massive chemical and biological diversity. Drugs for the treatment of skin and soft tissue infections have become part of the search, mainly with respect to enlarging the number of available antibiotics, with a special focus on multidrug-resistant Gram-positive bacteria, being the major causative agents in this field. Marine resources offer novel natural products with distinct biological activities of pharmaceutical importance, having the chance to provide new chemical scaffolds and new modes of action. New studies advance the field by proposing new strategies derived from an ecosystemic understanding for preventive activities against biofilms and new compounds suitable as disinfectants, which sustain the natural flora of the skin. Still, the development of new compounds is often stuck at the discovery level, as marine biotechnology also needs to overcome technological bottlenecks in drug development. This review summarizes its potential and shows these bottlenecks and new approaches.


Does improving appropriate use of malaria medicines change population beliefs in testing and treatment? Evidence from a randomized controlled trial.
A major puzzle in malaria treatment remains the dual problem of underuse and overuse of malaria medications, which deplete scarce public resources used for subsidies and lead to drug resistance. One explanation is that health behaviour, especially in the context of incomplete information, could be driven by beliefs, pivotal to the success of health interventions. The objective of this study is to investigate how population beliefs change in response to an experimental intervention which was shown to improve access to rapid diagnostic testing (RDT) through community health workers (CHWs) and to increase appropriate use of anti-malaria medications. By collecting data on individuals' beliefs on malaria testing and treatment 12 and 18 months after the experimental intervention started, we find that the intervention increases the belief that a negative test result is correct, and the belief that the first-line anti-malaria drugs (artemisinin-based combination therapies or ACTs) are effective. Using mediation analysis, we also explore some possible mechanisms through which the changes happen. We find that the experience and knowledge about RDT and experience with CHWs explain 62.4% of the relationship between the intervention and the belief that a negative test result is correct. Similarly, the targeted use of ACTs and taking the correct dose-in addition to experience with RDT-explain 96.8% of the relationship between the intervention and the belief that the ACT taken is effective. As beliefs are important determinants of economic behaviour and might guide individuals' future decisions, understanding how they change after a health intervention has important implications for long-term changes in population behaviour.


The insulin-like effect of sodium vanadate on adipocyte glucose transport is mediated at a post-insulin-receptor level.
Sodium vanadate has several insulin-like effects. To determine whether vanadate acts via the insulin receptor, I investigated the effect of vanadate on glucose transport (2-deoxyglucose uptake) in adipocytes that had been treated to decrease the number of insulin receptors. Trypsin (100 micrograms/ml) caused greater than 95% loss of 125I-insulin binding and rendered glucose transport resistant to both insulin and an anti-insulin-receptor antibody. However, vanadate caused an 8-fold increase in the transport rate [EC50 (concn. giving 50% of maximum effect) 0.2 mM] in both control and trypsin-treated cells, demonstrating that the insulin receptor does not have to be intact for vanadate to stimulate glucose transport. Insulin receptors were depleted by treatment of adipocytes with insulin (100 ng/ml) in the presence of Tris (which blocks receptor recycling). A 2 h treatment caused 60% loss of receptors, and a shift to the right in the dose-response curve for insulin stimulation of glucose transport (EC50 0.3 ng of insulin/ml in controls, 1.2 ng/ml in treated cells). The response to vanadate was again unaffected. Treatment with insulin for 4 h caused a 67% decrease in insulin binding and, in addition to the rightward shift in the insulin dose-response curve, a decrease in basal and maximal transport rates (which cannot be explained by decreased insulin receptor number). The EC50 of vanadate was again equal in control and treated cells, but glucose transport in the presence of a maximally effective concentration of vanadate (1 mM) was decreased. I conclude that the effect of vanadate on glucose transport is independent of the insulin receptor. Induction of a post-receptor defect (which may be a decrease in the total number of cellular glucose transporters) by prolonged exposure to insulin decreases the potency of a maximally effective concentration of vanadate. The findings demonstrate that vanadate stimulates glucose transport by an effect at a level distal to the insulin receptor.


Didanosine-loaded poly(epsilon-caprolactone) microparticles by a coaxial electrohydrodynamic atomization (CEHDA) technique.
The goal of this study was to investigate the electrohydrodynamic atomization (EHDA) technology to encapsulate the water-soluble antiretroviral didanosine (ddI) within poly(epsilon-caprolactone) (PCL) particles and stabilize it in the gastric medium where it undergoes fast degradation. A preliminary study employing a one-needle setup enabled the adjustment of the critical process parameters. Then, a configuration of two concentric needles named coaxial electrohydrodynamic atomization (CEHDA) led to the formation of ddI-loaded PCL microcapsules. Scanning electron microscopy analysis showed that the microparticles were spherical and with narrow size distribution. Attenuated total reflectance/Fourier transform infrared spectroscopy confirmed that most of the drug was efficiently encapsulated within the particles, whereas differential scanning calorimetry and X-ray powder diffraction revealed that the drug was preserved mainly in crystalline form. The loading capacity was relatively high (approximately 12% w/w), and the encapsulation efficiency was approximately 100%. In vitro release assays (PBS pH = 7.4) indicated that ddI was released almost completely within 2 h. Moreover, the delayed release was expected to isolate ddI from the biological fluids during the gastric transit. Finally, pharmacokinetics studies in rats showed that ddI-loaded particles lead to a statistically significant increase of the oral bioavailability of almost 4 times and a 2-fold prolongation of the half-life with respect to a ddI aqueous solution, supporting the use of CEHDA as a promising reproducible, scalable and cost-viable technology to encapsulate water-soluble drugs within polymeric particles.


The psychometric properties of the Chinese version of the Interoceptive Awareness Questionnaire (IAQ) among cardiovascular disease.
Interoceptive awareness (perception of body conditions and processes) and heart rate variability are connected physiologically in cardiovascular disease (CVD) patients. At present, there is no specific evaluation model for the perception of the physical state and processes of CVD patients in China. The objective of this study is to examine the reliability and validity of the Chinese Interoceptive Awareness Questionnaire (C-IAQ) for Chinese CVD patients. 160 CVD patients were recruited from a hospital in Hubei province using a convenient sampling method. A standard "forward-backward" translation method was applied to convert the C-IAQ into Mandarin. Split-half reliability and internal consistency were conducted by using reliability tests. Validity testing was conducted on the content, structure, and criterion-related validity. Criterion-related validity was assessed by using the Anxiety Sensitivity Index-III (ASI-III). The research results indicate that the dual factor structure of the original C-IAQ has 19 items, including attention to unpleasant sensations (9 items) and awareness of neutral body sensations (10 items). Moreover, C-IAQ is positively correlated with ASI-III (r = 0.48, P<0.01). The entire scale has a Cronbach's α value of 0.85 and split-half dependability of 0.77. The C-IAQ has favorable psychometric feature. Hence, it can be used to measure the interoceptive awareness of CVD patients.


Mouse Cntnap2 and Human CNTNAP2 ASD Alleles Cell Autonomously Regulate PV+ Cortical Interneurons.
Human mutations in CNTNAP2 are associated with an array of neuropsychiatric and neurological syndromes, including speech and language disorders, epilepsy, and autism spectrum disorder (ASD). We examined Cntnap2's expression and function in GABAergic cortical interneurons (CINs), where its RNA is present at highest levels in chandelier neurons, PV+ neurons and VIP+ neurons. In vivo functions were studied using both constitutive Cntnap2 null mice and a transplantation assay, the latter to assess cell autonomous phenotypes of medial ganglionic eminence (MGE)-derived CINs. We found that Cntnap2 constitutive null mutants had normal numbers of MGE-derived CINs, but had reduced PV+ CINs. Transplantation assays showed that Cntnap2 cell autonomously regulated the physiology of parvalbumin (PV)+, fast-spiking CINs; no phenotypes were observed in somatostatin+, regular spiking, CINs. We also tested the effects of 4 human CNTNAP2 ASD missense mutations in vivo, and found that they impaired PV+ CIN development. Together, these data reveal that reduced CNTNAP2 function impairs PV+ CINs, a cell type with important roles in regulating cortical circuits.


Effect of chronic renal insufficiency on outcomes of carotid endarterectomy.
Conflicting data exist regarding the effect of chronic renal insufficiency (CRI) on carotid endarterectomy (CEA) outcomes. A large database was used to analyze the effect of CRI, defined by glomerular filtration rate (GFR), as an independent risk factor of CEA. Prospectively collected data regarding CEAs performed at 123 Veterans Affairs Medical Centers as part of the National Surgical Quality Improvement Program were retrospectively analyzed. Renal function was used to divide patients into three CRI groups: normal or mild (control; GFR >/=60 mL/min/1.73 m(2)), moderate (GFR 30 to 59), and severe (GFR <30). Bivariate analysis and multivariate logistic regression were used to characterize risk factors and their associations with 30-day morbidity and mortality. Between Jan 1, 1996, and Dec 31, 2003, 22,080 patients underwent CEA. Patients missing creatinine levels, already dialysis-dependent, or in acute renal failure just before surgery were excluded. This left 20,899 available for analysis, of which 13,965 had a GFR of >/=60, 6,423 had a GFR of 30 to 59, and 511 had a GFR of <30. The incidence of neurologic complications did not differ significantly (control, 1.7%; moderate CRI, 1.9%; severe CRI, 2.7%). The moderate CRI group experienced significantly more cardiac events (1.7% vs 0.9% for controls, P < .001). This remained predictive in the multivariate model even adjusting for all other risk factors (adjusted odds ratio [AOR], 1.6; 95% confidence interval [CI], 1.1-2.3; P = .009). The moderate CRI group also had higher rates of pulmonary complications (2.1% vs 1.3% control; P < .001; AOR, 1.3; 95% CI, 1.0-1.7; P = .031) but not 30-day mortality (P = .269). Those with severe CRI had a much higher mortality (3.1% vs 1.0% control, P < .001), which remained significant in the multivariate model (AOR, 2.7; 95% CI, 1.6-4.8; P < .001). Although impaired renal function does not independently increase the risk of neurologic or infectious complications, CRI is a significant negative independent risk factor in predicting other outcomes after CEA. Patients with moderate CRI (GFR, 30-59 mL/min/1.73 m(2)) are at increased risk for cardiac and pulmonary morbidity, but not death, and those with severe CRI (GFR <30 mL/min/1.73 m(2)) have a much higher operative mortality. Patients with CRI should be carefully evaluated before CEA to optimize existing cardiac and pulmonary disease. Understanding this increased risk may assist the surgeon in preoperative counseling and perioperative management.


The embodiment of lyricism in medicine and Homer.
Improving the quality of communication between doctors and their patients and colleagues is of vital importance. Poor communication, especially within and across clinical teams working around patients in pathways of care, leads to avoidable medical error, where an unacceptable number of patients are severely harmed or die each year. The figures from such iatrogenesis have now reached epidemic proportions, constituting one of the major killers of patients worldwide. Despite 30 years' worth of explicit attention to teaching communication skills at undergraduate level, communication in medicine is failing to improve at an acceptable rate. The authors suggest a rather unusual approach to this dilemma of 'communication hypocompetence'—thinking medicine lyrically—as an extension of thinking with Homer's little-discussed lyrical aesthetic. A key part of the problem of communication hypocompetence is the well-researched phenomenon of 'empathy decline' in students, where 'hardening' and cynicism occur as over-determined ego defences. Empathy decline may be a symptom of the repression of the lyrical genre in medicine, where the epic, tragic and dark comic genres dominate. The lyrical genre emphasises coming to know the patient as a person and an individual. Importantly, central to performing the lyric genre is the heightened use of the senses in taking a history, physical examination and diagnostic work. Framing medicine as lyrical work challenges undue emphasis on 'cure' at the expense of humane 'care'.


Transcriptome dynamics of the microRNA inhibition response.
We report a high-resolution time series study of transcriptome dynamics following antimiR-mediated inhibition of miR-9 in a Hodgkin lymphoma cell-line-the first such dynamic study of the microRNA inhibition response-revealing both general and specific aspects of the physiological response. We show miR-9 inhibition inducing a multiphasic transcriptome response, with a direct target perturbation before 4 h, earlier than previously reported, amplified by a downstream peak at ∼32 h consistent with an indirect response due to secondary coherent regulation. Predictive modelling indicates a major role for miR-9 in post-transcriptional control of RNA processing and RNA binding protein regulation. Cluster analysis identifies multiple co-regulated gene regulatory modules. Functionally, we observe a shift over time from mRNA processing at early time points to translation at later time points. We validate the key observations with independent time series qPCR and we experimentally validate key predicted miR-9 targets. Methodologically, we developed sensitive functional data analytic predictive methods to analyse the weak response inherent in microRNA inhibition experiments. The methods of this study will be applicable to similar high-resolution time series transcriptome analyses and provides the context for more accurate experimental design and interpretation of future microRNA inhibition studies.


Remuneration, freedom of choice, professional guinea pig.
The law of December 20, 1988 states that a participant in a clinical test without direct therapeutic benefit is not entitled to receive remuneration. Professionalism may be the strongest reason for participating. The law prohibits some participants from receiving an indemnity, not on the basis of healthy vs sick discrimination but when a situation of particular vulnerability or dependence has been established.


Efficiency of the health extension programme in Tigray, Ethiopia: a data envelopment analysis.
Since 2004, the government of Ethiopia has made a bold decision to strengthen and expand its primary health care system by launching the Health Extension Program (HEP). While the scaling up of the HEP is necessary to achieve the aim of universal access to primary health care, close attention should be paid to the performance of the program. Using a data envelopment analysis this study aimed at (i) to estimate the technical efficiency of a sample of health posts in rural Tigray, ii) to identify those factors which might be explaining the efficiency results. Efficiency was measured using a data envelopment analysis model. A Tobit model was performed to identify factors associated with efficiency. Seven rural districts (out of 35) were purposely chosen. Input/output information was collected from the database of the Tigray Health Bureau during July 2007-June 2008. Information was also collected on environmental factors that might influence the efficiency outcomes through a structured questionnaire from the correspondent district health officers. Analysis was based on data from 60 health posts. The mean scores for technical and scale efficiency were 0.57 (SD = 0.32) and 0.95 (SD = 0.11) respectively. Out of the 60 health posts, 15 (25.0%) were found to be technically efficient constituting the best practice frontier. Thirty eight (63.3%) were operating at their most productive scale size. In the regression analysis, none of the variables was significantly associated with the efficiency outcome. There is a need to review the management of the health information system in the region. The findings have also revealed that only a quarter of the health posts are working efficiently and pointed the need for improvement. A closer monitoring of the health extension programme is required in order to achieve the best possible performance.


Older people living alone at home with terminal cancer.
This study describes the lived experiences of older people coping with terminal cancer and living alone, focusing on how they face challenges of the biographical life changes from their disease progression. Face-to-face semi-structured interviews were conducted in two phases with palliative care clients of a community-based service in Western Australia (2009-2011): Brief interviews with 43 cancer patients who live alone and then in-depth interviews with 8 of them. Using biographical disruption as the analytical framework for interpreting the qualitative data, four main themes emerged: Biographical disruption: adjusting to change; Biographical continuity: preserving normality; Biographical reconstruction: redefining normality; and Biographical closure: facing the end. Biographical disruption was a suitable framework for analysis, permitting identification of the biographical disruptions of the individual's world and the reframing that is undertaken by the individual to maintain autonomy and independence while acknowledging and accepting their closeness to death. Understanding the factors associated with the individual's need to maintain their own identity will enable nurses working with this population to tailor support plans that meet the individuals' needs while maintaining or restoring the person's sense of self. Interventions that directly address end-of-life suffering and bolster sense of dignity and personhood need to be considered.


Cobinamide-based cyanide analysis by multiwavelength spectrometry in a liquid core waveguide.
A novel cyanide analyzer based on sensitive cobinamide chemistry relies on simultaneous reagent and sample injection and detection in a 50 cm liquid core waveguide (LCW) flow cell illuminated by a white light emitting diode. The transmitted light is read by a fiber-optic charge coupled device (CCD) spectrometer. Alkaline cobinamide (orange, lambda(max) = 510 nm) changes to violet (lambda(max) = 583 nm) upon reaction with cyanide. Multiwavelength detection permits built-in correction for artifact responses intrinsic to a single-line flow injection system and corrects for drift. With optimum choice of the reaction medium, flow rate, and mixing coil length, the limit of detection (LOD, S/N = 3) is 30 nM and the linear dynamic range extends to 10 microM. The response base width for 1% carryover is <95 s, permitting a throughput of 38 samples/h. The relative standard deviations (rsd) for repetitive determinations at 0.15, 0.5, and 1 microM were 7.6% (n = 5), 3.2% (n = 7), and 1.7% (n = 6), respectively. Common ions at 250-80,000x concentrations do not interfere except for sulfide. For the determination of 2 microM CN(-), the presence of 2, 5, 10, 20, 100, and 1000 microM HS(-) results in 22, 27, 48, 58, 88, and 154% overestimation of cyanide. The sulfide product actually has a different characteristic absorption, and in those samples where significant presence is likely, this can be corrected for. We demonstrate applicability by analyzing the hydrolytic cyanide extract of apple and pear seeds with orange seeds as control and also measure HCN in breath air samples. Spike recoveries in these sample extracts ranged from 91 to 108%.


A metabolomic comparison of mouse models of the Neuronal Ceroid Lipofuscinoses.
The Neuronal Ceroid Lipofuscinoses (NCL) are a group of fatal inherited neurodegenerative diseases in humans distinguished by a common clinical pathology, characterized by the accumulation of storage body material in cells and gross brain atrophy. In this study, metabolic changes in three NCL mouse models were examined looking for pathways correlated with neurodegeneration. Two mouse models; motor neuron degeneration (mnd) mouse and a variant model of late infantile NCL, termed the neuronal ceroid lipofuscinosis (nclf) mouse were investigated experimentally. Both models exhibit a characteristic accumulation of autofluorescent lipopigment in neuronal and non neuronal cells. The NMR profiles derived from extracts of the cortex and cerebellum from mnd and nclf mice were distinguished according to disease/wildtype status. In particular, a perturbation in glutamine and glutamate metabolism, and a decrease in γ-amino butyric acid (GABA) in the cerebellum and cortices of mnd (adolescent mice) and nclf mice relative to wildtype at all ages were detected. Our results were compared to the Cln3 mouse model of NCL. The metabolism of mnd mice resembled older (6 month) Cln3 mice, where the disease is relatively advanced, while the metabolism of nclf mice was more akin to younger (1-2 months) Cln3 mice, where the disease is in its early stages of progression. Overall, our results allowed the identification of metabolic traits common to all NCL subtypes for the three animal models.


Active intervention in hospital test request panels pays.
Ordering laboratory tests by means of test panels is a convenient way of requesting tests, preventing necessary tests from being forgotten. However, it also leads to redundant test ordering, as not all tests in a given panel are required for each patient. As test panels proposed by doctors may contain redundant, overlapping or infrequently used tests, the active involvement of knowledgeable laboratory staff in the organisation of test panels is advisable to promote efficient test use. Laboratory staff initiated an intervention in the organisation of test panels at our hospital in 2009. After a review of the existing panels and the proposals for new panels, we established a total of 60 panels (down from 171 previously). We also stipulated that the laboratory is to be involved with all proposals for new test panels in the future. The reorganisation reduced the number of tests in the test panels by 17.7% (n=60), which theoretically should have resulted in 4.5% fewer tests being ordered. However, as an estimated 14% of the tests removed were then ordered individually in addition to the panels, 3.9% fewer tests were ordered, yielding an annual saving of about €58,000 (4.5% of the costs of all tests ordered in test panels). The savings amount to 7-8% if the frequently ordered metabolic panel (which was left unchanged) was excluded from the survey. Active intervention by the laboratory in the organisation of test panels results in a reduction in the use of tests and in interesting savings.


In vitro and in vivo tests for the biological evaluation of candidate orthopedic materials: Benefits and limits.
The development of reliable experimental models for the clinical use of biomaterials and in predicting implant success or failure is becoming increasingly important in attaining adequate health and safety conditions. This paper summarizes the main benefits and limits of in vitro and in vivo biological tests taking into account general pathophysiological considerations regarding the skeletal tissue. Current findings demonstrate the need to adopt both in vitro and in vivo methodologies and to improve test quality with particular attention to the ethical issues associated with in vivo experiments. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 155-63).


A de novo expression profiling of Anopheles funestus, malaria vector in Africa, using 454 pyrosequencing.
Anopheles funestus is one of the major malaria vectors in Africa and yet there are few genomic tools available for this species compared to An. gambiae. To start to close this knowledge gap, we sequenced the An. funestus transcriptome using cDNA libraries developed from a pyrethroid resistant laboratory strain and a pyrethroid susceptible field strain from Mali. Using a pool of life stages (pupae, larvae, adults: females and males) for each strain, 454 sequencing generated 375,619 reads (average length of 182 bp). De novo assembly generated 18,103 contigs with average length of 253 bp. The average depth of coverage of these contigs was 8.3. In total 20.8% of all reads were novel when compared to reference databases. The sequencing of the field strain generated 204,758 reads compared to 170,861 from the insecticide resistant laboratory strain. The contigs most differentially represented in the resistant strain belong to the P450 gene family and cuticular genes which correlates with previous studies implicating both of these gene families in pyrethroid resistance. qPCR carried out on six contigs indicates that these ESTs could be suitable for gene expression studies such as microarray. 31,000 sites were estimated to contain Single Nucleotide Polymorphisms (SNPs) and analysis of SNPs from 20 contigs suggested that most of these SNPs are likely to be true SNPs. Gene conservation analysis confirmed the close phylogenetic relationship between An. funestus and An. gambiae. This study represents a significant advance for the genetics and genomics of An. funestus since it provides an extensive set of both Expressed Sequence Tags (ESTs) and SNPs which can be readily adopted for the design of new genomic tools such as microarray or SNP platforms.


Immunoelectrophoresis of Mycobacterium tuberculosis antigens. Comparative analysis of cell extract and culture filtrate antigens.
Immunoelectrophoretic analyses were performed on a cell extract and culture filtrate of Mycobacterium tuberculosis, H37Rv strain using homologous polyvalent goat antisera. General similarity between the cell extract and culture filtrate antigens was found. Significant variation between two anti-culture filtrate antisera was observed. These antigens and antisera also differed significantly from previously described reference reagents. These findings demonstrate the necessity of using reference materials for precise identification of mycobacterial antigens in comparative studies.


Trends in the incidence of cutaneous malignant melanoma in New South Wales, 1983-1996.
The incidence of cutaneous malignant melanoma (CMM) has been rising in fair-skinned populations throughout the world for decades. The upward trend may, however, finally be slowing in some of these populations. Recent (1983-1996) CMM incidence trends for a high incidence area (New South Wales, Australia) have been examined according to gender, age group, body site and tumour thickness. Despite continuing upward trends in older age groups, particularly among men (e.g., 7.20% increase per year in men aged 75+), incidence for younger ages is stabilizing (in men) or declining (in women): average annual percentage changes of -3.03 and -0.88 were observed for women aged 15-34 and 35-54, respectively. Patterns suggest a birth-cohort effect, with those born since 1945 or 1950 having lower (females) or similar (males) rates to those born earlier. For each gender, all-ages incidence rose by a similar amount for each of the main body sites except the leg in women, where incidence fell by 0.49% per year. In men, the incidence of both thin (</=75 mm) and thick (>75 mm) melanomas increased (significantly, by 2.63% per year and non-significantly, by 0.93% per year, respectively) between 1989 and 1996. In women, incidence remained stable for both thickness subgroups. These data are consistent with a stabilization or reduction in either total sun exposure or intermittency of exposure among New South Wales cohorts born since about 1950. Because incidence rates are still much higher than they were a few decades ago, however, efforts to reduce sun exposure, particularly in children and youth, must continue.


Carthamus Tınctorius L. (Safflower) extracts inhibit expression of metastatic genes of MDA-MB-231 breast cancer cells.
Breast cancer is the most common type of cancer in women and the second cause of cancer-related death after lung cancer. Although the common methods used in the treatment of breast cancer are chemotherapy, radiotherapy and surgery, the search for alternative treatments continues. The leading alternative treatments are medicinal plants which actually inspire the production of many cancer drugs. In this study, the proliferative and metastatic effects of Carthamus tinctorius L., known for its many therapeutic properties, on metastatic breast cancer were investigated. Here, intending to evaluate the the content and actions of different extracts of safflower leaves extracts were prepared by extracting in water, alcohol and oil and analysed by FTIR. Their antioxidant effect was tested and then the extracts were applied to metastatic breast cancer cells. FTIR spectrums of all three extracts have revealed the presence of organic compounds. It is found that all extracts but mostly the oil extract has antioxidant property. MTT assay, wound healing assay and gene expression analysis were performed to assess the antiproliferative and anti metastatic effects of the extracts on breast cancer cells. It is found that, there is no significant antiproliferative effect of extracts on MDA-MB-231 cells except the alcohol extract. However, all safflower extracts, especially the oil extract, significantly reduced the metastatic potential of breast cancer cells. It is concluded that safflower contents are potent chemicals which inhibit the cellular mechanisms underlying the spreading of cancer cells and further analysis may lead to new initiatives in drug design research.


One year survival of HIV-infected veterans with CD4 < 100 cells/mm3: data from a veteran cohort.
Survival among HIV-infected patients markedly improved with the introduction of highly active antiretroviral therapy (HAART). Easier to take and more effective HAART options have improved the one-year virologic success rate among naive patients. Numerous studies have shown that initiating HAART and restoration of CD4 cells positively impact survival. There are only a few evaluations that have been carried out on the changes in survival among patients who are severely immunosuppressed. We evaluated survival among a cohort of veterans with CD4<100 cells/mm(3) (CD4 < 100) in three time periods reflecting early, mid, and recent HAART. Using the HIV clinic database, all patients with CD4 < 100 seen between 1996 and 2004 were identified (n=394). Patients entered Cohorts 1 (n=219), 2 (n=72), and 3 (n=103) in 1996-1998, 1999-2001, and 2002-2004, respectively. Data on demographics, AIDS-defining illnesses, co-morbidities, treatment, CD4, and viral load (VL) were abstracted. Survival analysis controlling for the above variables was performed and odds ratios with 95% confidence intervals were calculated. Rate of virologic suppression was higher for Cohort 2 when compared to Cohort 1 (63% vs. 46%, p<0.05), but lower for Cohort 3 when compared to Cohort 2 (49%, p<0.05). Survival at one year was high for Cohorts 1 and 2 (92-95%), but significantly lower in Cohort 3 (80%). On logistic regression analysis and for the whole cohort, HAART use, achieving a CD4 > 200 and VL<400 were independent predictors of survival. Older age at cohort entry and having a diagnosis of lymphoma, Mycobacterium avium complex infection, coronary artery disease, or renal insufficiency were negative predictors. In the most recent HAART period 2002-2004, one year survival after CD4 < 100 significantly decreased in spite of availability of specialized HIV clinical and support services and antiretrovirals. Our results suggest that more than better drugs are needed for improving survival among certain patient populations with advanced immunosuppression.


Skeletal muscle telomere length in healthy, experienced, endurance runners.
Measuring the DNA telomere length of skeletal muscle in experienced endurance runners may contribute to our understanding of the effects of chronic exposure to endurance exercise on skeletal muscle. This study compared the minimum terminal restriction fragment (TRF) length in the vastus lateralis muscle of 18 experienced endurance runners (mean age: 42 +/- 7 years) to those of 19 sedentary individuals (mean age: 39 +/- 10 years). The runners had covered almost 50,000 km in training and racing over 15 years. Minimum TRF lengths measured in the muscle of both groups were similar (P = 0.805) and within the normal range. Minimum TRF length in the runners, however, was inversely related to their years spent running (r = -0.63, P = 0.007) and hours spent training (r = -0.52, P = 0.035). Therefore, since exposure to endurance running may influence minimum TRF length, and by implication, the proliferative potential of the satellite cells, chronic endurance running may be seen as a stressor to skeletal muscle.


Mitogens and oncogenes can block the induction of specific voltage-gated ion channels.
The mechanisms underlying the ontogeny of voltage-gated ion channels in muscle are unknown. Whether expression of voltage-gated channels is dependent on mitogen withdrawal and growth arrest, as is generally true for the induction of muscle-specific gene products, was investigated in the BC3H1 muscle cell line by patch-clamp techniques. Differentiated BC3H1 myocytes expressed functional Ca2+ and Na+ channels that correspond to those found in T tubules of skeletal muscle. However, Ca2+ and Na+ channels were first detected after about 5 days of mitogen withdrawal. In order to test whether cellular oncogenes, as surrogates for exogenous growth factors, could prevent the expression of ion channels whose induction was contingent on mitogen withdrawal, BC3H1 cells were modified by stable transfection with oncogene expression vectors. Expression vectors containing v-erbB, or c-myc under the control of the SV40 promoter, delayed but did not prevent the appearance of functional Ca2+ and Na+ channels. In contrast, transfection with a Val12 c-H-ras vector, or cotransfection of c-myc together with v-erbB, suppressed the formation of functional Ca2+ and Na+ channels for greater than or equal to 4 weeks. Potassium channels were affected neither by mitogenic medium nor by transfected oncogenes. Thus, the selective effects of certain oncogenes on ion channel induction corresponded to the suppressive effects of mitogenic medium.


[Precision diagnostics and therapy in hematological malignancies].
Precision diagnostics and therapy have been implemented rather early in clinical hematology due to the easy accessibility of blood and bone marrow, allowing not only for consecutive genetic analysis at diagnosis, remission and relapse, but also for culturing these cells and testing new drugs in vitro. One contributing factor has also been the relatively low number of »driver« mutations in hematologic malignancies and that some of them are gain of function mutations that are relatively easy to target by drugs. Examples of this development are ABL1-, JAK2-, and FLT3-inhibitors for the treatment of chronic myeloid leukemia, myeloproliferative neoplasms, and acute myeloid leukemia, respectively. More recently, gene panel sequencing has been introduced in clinical routine to identify genetic alterations with diagnostic, prognostic and predictive impact, and more sensitive techniques to monitor minimal residual disease are emerging. Whole genome and transcriptome sequencing are currently evaluated as the next diagnostic tool. Finally, a large number of targeted therapies are currently under development and/or undergoing clinical trials.


Purification and characterization of neutral and alkaline invertase from carrot.
Neutral and alkaline invertase were identified in cells of a suspension culture of carrot (Daucus carota L.) and purified to electrophoretic homogeneity. Neutral invertase is an octamer with a molecular mass of 456 kD and subunits of 57 kD, whereas alkaline invertase is a tetramer with a molecular mass of 504 kD and subunits of 126 kD. Both enzymes had sharp pH profiles, with maximal activities at pH 6.8 for neutral invertase and pH 8.0 for alkaline invertase, and both hydrolyzed sucrose with typical hyperbolic kinetics and similar Km values of about 20 mM at pH 7.5. Neutral invertase also hydrolyzed raffinose and stachyose and, therefore, is a beta-fructofuranosidase. In contrast, alkaline invertase was highly specific for sucrose. Fructose acted as a competitive inhibitor of both enzymes, with Ki values of about 15 mM. Glucose was a noncompetitive inhibitor of both neutral and alkaline invertase, with a Ki of about 30 mM. Neither enzyme was inhibited by HgCl2. Alkaline invertase was markedly inhibited by CaCl2, MgCl2, and MnCl2, and neutral invertase was not. In contrast to alkaline invertase, neutral invertase was inhibited by the nucleotides ATP, CTP, GTP, and UTP.


Is 3D ultrasound reliable for the evaluation of carotid disease? A systematic review and meta-analysis.
Studies assessing the use of 3D ultrasound (3DUS) for the evaluation of carotid disease reported varying views among observers about its reliability vis-à-vis 2DUS or angiography; ratings provided ranged from poor to excellent. Thisstudy aims to systematically review and analyze the reliability of 3DUS for the evaluation of carotid disease. The PubMed database was searched for studies that evaluated carotid disease (i.e. plaque measurements and characteristics and degree of stenosis) using 3DUS. Sixteen studies comprising a total of 918 stenosed carotids were reviewed and meta-analyzed. Data on intra- and inter-observer reproducibility and inter-method agreement (i.e. 3DUS vs 2D and 3DUS vs angiography) were analyzed. Overall analysis showed excellent intra- and inter-observer reproducibility (intraobserver: correlation coefficient r=0.88, 95% confidence intervals (CI) 0.84-0.92; intra-observer: r=0.91, 95% CI 0.87-0.95). The analysis also showed excellent agreement between 3DUS and 2DUS (r=0.89, 95% CI 0.83-0.95) and between 3DUS and angiography (r=0.73, 95% CI 0.44-0.1). 3DUS has excellent intra- and inter-observer reproducibility and excellent agreement with 2DUS and angiography for the evaluation of carotid disease. Further studies assessing the reliability ofcarotid plaque characteristics using 3DUS in symptomatic and asymptomatic patients are required.


[Hematopoietic changes in mice with transplantable Lewis lung carcinoma].
The cell composition of bone marrow, spleen, and peripheral blood as well as the content of early granulocytic and macrophagal precursor cells forming colonies in agar cultures in diffusion chambers in vivo (CFU-DC) were studied in C57B1/6 mice during the Lewis carcinoma growth after subcutaneous transplantation of tumour cells. It has been shown that in peripheral blood of such mice neutrophilic leucocytosis develops and the content of CFU-DC increases considerably. The spleen has become a source of surplus production of granulocytic and macrophagal cells from the early precursors (CFU-DC) to the mature cell forms with their delivery to the peripheral blood. No essential changes in the cell composition and CFU-DC content were found in the bone marrow of femur.


Quality of perioperative AEP--variability of expert ratings.
Previous studies suggest that auditory evoked potentials (AEP) may be used to monitor anaesthetic depth. However, during surgery and anaesthesia, the quality of AEP recordings may be reduced by artefacts. This can affect the interpretation of the data and complicate the use of the method. We assessed differences in expert ratings of the signal quality of perioperatively recorded AEPs. Signal quality of 180 randomly selected AEP, recorded perioperatively during a European multicentre study, was rated independently by five experts as 'invalid' (0), 'poor' (1), or 'good' (2). Average (n=5) quality rating was calculated for each signal. Differences between quality ratings of the five experts were calculated for each AEP: inter-rater variability (IRV) was calculated as the difference between the worst and best classification of a signal. Average signal quality of 57% of the AEPs was rated as 'invalid', 39% as 'poor', and only 4% as 'good'. IRV was 0 in only 6%, 1 in 62%, and 2 in 32% of the AEP, that is in 32% one expert said signal quality was good, whereas a different expert thought the identical signal was invalid. There is poor agreement between experts regarding the signal quality of perioperatively recorded AEPs and, as a consequence, results obtained by one expert may not easily be reproduced by a different expert. This limits the use of visual AEP analysis to indicate anaesthetic depth and may affect the comparability of AEP studies, where waveforms were analysed by different experts. An objective automated method for AEP analysis could solve this problem.


Conformational Shift of a β-Hairpin Peptide upon Complex Formation with an Oligo-proline Peptide Studied by Mass Spectrometry.
So-called super-secondary structures such as the β-hairpin, studied here, form an intermediate hierarchy between secondary and tertiary structures of proteins. Their sequence-derived 'pure' peptide backbone conformation is combined with 'remote' interstrand or interresidue contacts reminiscent of the 3D-structure of full-length proteins. This renders them ideally suited for studying potential nucleation sites of protein folding reactions as well as intermolecular interactions. But β-hairpins do not merely serve as model systems; their unique structure characteristics warrant a central role in structural studies on their own. In this study we applied photo cross-linking in combination with high-resolution mass spectrometry and computational modeling as well as with ion mobility-mass spectrometry to elucidate these structural properties. Using variants of a known β-hairpin representative, the so-called trpzip peptide and its ligands, we found evidence for a conformational transition of the β-hairpin and its impact on ligand binding.


Altered profile of mRNA expression in atrioventricular node of streptozotocin‑induced diabetic rats.
Prolonged action potential duration, reduced action potential firing rate, upstroke velocity and rate of diastolic depolarization have been demonstrated in atrioventricular node (AVN) cells from streptozotocin (STZ)‑induced diabetic rats. To further clarify the molecular basis of these electrical disturbances, the mRNA profiles encoding a variety of proteins associated with the generation and conduction of electrical activity in the AVN, were evaluated in the STZ‑induced diabetic rat heart. Expression of mRNA was measured in AVN biopsies using reverse transcription‑quantitative polymerase chain reaction techniques. Notable differences in mRNA expression included upregulation of genes encoding membrane and intracellular Ca2+ transport, including solute carrier family 8 member A1, transient receptor potential channel 1, ryanodine receptor 2/3, hyperpolarization‑activated cyclic‑nucleotide 2 and 3, calcium channel voltage‑dependent, β2 subunit and sodium channels 3a, 4a, 7a and 3b. In addition to this, potassium channels potassium voltage‑gated channel subfamily A member 4, potassium channel calcium activated intermediate/small conductance subfamily N α member 2, potassium voltage‑gated channel subfamily J members 3, 5, and 11, potassium channel subfamily K members 1, 2, 3 and natriuretic peptide B (BNP) were upregulated in AVN of STZ heart, compared with controls. Alterations in gene expression were associated with upregulation of various proteins including the inwardly rectifying, potassium channel Kir3.4, NCX1 and BNP. The present study demonstrated notable differences in the profile of mRNA encoding proteins associated with the generation, conduction and regulation of electrical signals in the AVN of the STZ‑induced diabetic rat heart. These data will provide a basis for a substantial range of future studies to investigate whether variations in mRNA translate into alterations in electrophysiological function.


[The immune system, atherosclerosis and persisting infection].
The paper demonstrates that lymph nodes situated in the vicinity of magistral blood vessels are the source of immune and inflammatory response to LDL as the main pathogenetic factor in atherosclerosis. The activation of T-cell-mediated immunity takes place in them at the very early stages of the disease, resulting in forming of CD4+T-lymphocytes, activated mononuclear cells and immunostabilizing B-lymphocytes. The cell changes in lymph nodes correlate with the severity of atherosclerotic lesions in the vessel intima and closely reflect the peculiarities of immune inflammation development in fatty streaks and atherosclerotic plaques in human atherogenesis. A paradoxical reaction was observed in cases with Chlamydia pneumoniae found in the wall of aorta and paraaortal lymph nodes. No evident immune response on the part of immunocompetent cells took place, but, on the contrary, the function of mononuclear cells, including T-lymphocytes, was suppressed. This phenomenon may be explained by the fact that intracellular localization of Chlamydia pneumoniae hides it from immune system control or by the possible microorganism capability of direct immunosuppressive influence on lymphoid cells both in the blood vessel wall and in regional lymph nodes.


Effects of ganopoly (a Ganoderma lucidum polysaccharide extract) on the immune functions in advanced-stage cancer patients.
Preclinical studies have established that the Ganoderma lucidum polysaccharide (GLPS) fractions have potent anti-tumor activity, which has been associated with the immuno-stimulating effects of GLPS. However, it is unclear whether GLPS has immuno-modulating effects in humans in vivo. This study aimed to investigate the effects of Ganopoly, the polysaccharides fractions extracted from G. lucidum, on the immune function of advanced-stage cancer patients. Thirty-four advance-stage cancer patients were entered onto this study, and treated with 1800 mg Ganopoly, three times daily orally before meals for 12 weeks. Immune parameters (cytokines, T cell subsets, mitotic response to phytohemagglutinin (PHA) and natural killer activity) were compared between baseline and after 12-week treatment. Thirty patients are assessable for their immune functions. Treatment of Ganopoly for 12 weeks resulted in a significant (P < 0.05) increase in the mean plasma concentrations of interleukin (IL-2), IL-6, and interferon (IFN)-gamma, whereas the levels of IL-1 and tumor necrosis factor (TNF-alpha) were significantly (P < 0.05) decreased. A marked variability among patients with advanced-stage cancer was observed in the numbers of each lymphocyte subset at baseline. The mean absolute number of CD56+ cells was significantly (P < 0.05) increased after 12-week treatment of Ganopoly, whereas the numbers of CD3+, CD4+, and CD8+ were just marginally increased compared to baseline levels, with the CD4:CD8 T cell ratios unchanged. PHA responses after 12-week treatment with Ganopoly were enhanced in most patients, when compared to pretreatment baselines (P < 0.05). In addition, Ganopoly treatment resulted in a significant increase (P < 0.05) in the mean NK activity compared to baselines (34.5 +/- 11.8% vs 26.6 +/- 8.3%). The present study indicates that Ganopoly enhanced the immune responses in patients with advanced-stage cancer. Clinical evaluations of response and toxicity are ongoing.


Influence of partial liquid ventilation on bacterial growth and alveolar expansion in newborn rabbits with group B-streptococcal pneumonia.
Partial liquid ventilation (PLV) with perfluorocarbons has been considered as an alternative therapy for severe inflammatory lung disease. The present study was performed to test whether PLV influences bacterial growth and lung histology in a rabbit model of congenital pneumonia caused by group B streptococci. Near-term newborn rabbits were tracheotomized, inoculated via the airways with group B streptococci, and subsequently ventilated for 5 h with either PLV or conventional ventilation. At 30 min after group B streptococci administration, animals in the PLV group (n = 16) received 30 mL/kg body weight of perfluorocarbon (PF 5080) via the tracheal tube. Evaporative losses were substituted with 20 mL/kg perfluorocarbon at hourly intervals. Identical volumes of air were injected in control animals at the same times (n = 15). The number of colony-forming units in left lung homogenate, evaluated at the end of the experiments, tended to be lower in PLV-treated animals than in controls (6.8 x 109 versus 6.4 x 1010 colony-forming units/g body weight; p = 0.06). Comparison of these numbers with the colony-forming units injected at the beginning of the experiments revealed a reduction in bacterial number in the PLV group and proliferation in the controls (-2.2 x 108 versus +5.6 x 1010 colony-forming units/g body weight; p < 0.05). Histologic examination demonstrated less inflammation and more homogeneous lung expansion in PLV-treated animals. Two animals in the PLV group had focal interstitial emphysema. Our results suggest that PLV with PF 5080 reduces bacterial proliferation in experimental group B streptococcal pneumonia.


Detection of Borrelia miyamotoi and other tick-borne pathogens in human clinical specimens and Ixodes scapularis ticks in New York State, 2012-2015.
Borrelia miyamotoi (Bm) is a recently emerging bacterial agent transmitted by several species of ixodid ticks. Diagnosis of Bm infection can be challenging, as the organism is not easily cultivable. We have developed and validated a multiplex real-time PCR to simultaneously identify Bm infection and the agents causing human granulocytic anaplasmosis and human monocytic ehrlichiosis, Anaplasma phagocytophilum and Ehrlichia chaffeensis, respectively. The assay is 100% specific; highly sensitive, detecting 11 gene copies of Bm DNA in both whole blood and cerebral spinal fluid; and provides rapid results in less than two hours. A retrospective study of 796 clinical specimens collected between the years 2012 and 2014 and a prospective study of 366 clinical specimens were performed utilizing this novel assay to evaluate the frequency of Bm infection in New York State (NYS). Eight clinical specimens (1%) were found to be positive for Bm, 216 were positive for A. phagocytophilum, and 10 were positive for E. chaffeensis. Additionally, we tested 411 I. scapularis ticks collected in NYS during 2013 and 2014 in a separate multiplex real-time PCR to determine the prevalence of Bm, A. phagocytophilum, Borrelia burgdorferi s.s., and Borrelia species. Our results indicated rates of 1.5%, 27%, 19.7%, and 8.8% respectively. The ability to monitor both the frequency and geographic distribution of Bm cases and the prevalence and geographic distribution of Bm in ticks will help create a better understanding of this emerging tick-borne pathogen.


Biomarkers and receptor targeted therapies reduce clinical trial risk in non-small-cell lung cancer.
This study analyzed the risk of clinical trial failure during non-small-cell lung cancer (NSCLC) drug development between 1998 and January 2012. We also looked for factors that impacted clinical trial risk in NSCLC. NSCLC drug development was investigated using trial disclosures from http://www.clinicaltrials.gov and other publically available resources. Compounds were excluded from the analysis if they had begun phase I clinical testing before 1998, did not use treatment-relevant endpoints, or if they did not have a completed phase I trial in NSCLC. Analysis was conducted in regard to treatment indication, compound classification, and mechanism of action. Six hundred seventy-six clinical trials that included 199 unique compounds met our inclusion criteria. The likelihood, or cumulative clinical trial success rate, that a new drug would pass all phases of clinical testing and be approved was found to be 11%, which is less than industry aggregate rates. Over half of the biomarkers used in NSCLC have not yet been approved by the Food and Drug Administration in any indication. Biomarker targeted therapies (62%) and receptor targeted therapies (31%) were found to have the highest success rates. The risk-adjusted cost for NSCLC clinical drug development was calculated to be U.S. $1.89 billion. Biomarker use alone in this indication resulted in a sixfold increase in clinical trial success whereas receptor targeted therapies did so by almost threefold. Physicians who enroll patients in NSCLC trials should prioritize their participation in clinical trial programs that use biomarkers and receptor targeted therapies.


Relating the physical properties of Pseudomonas aeruginosa lipopolysaccharides to virulence by atomic force microscopy.
Lipopolysaccharides (LPS) are an important class of macromolecules that are components of the outer membrane of Gram-negative bacteria such as Pseudomonas aeruginosa. P. aeruginosa contains two different sugar chains, the homopolymer common antigen (A band) and the heteropolymer O antigen (B band), which impart serospecificity. The characteristics of LPS are generally assessed after isolation rather than in the context of whole bacteria. Here we used atomic force microscopy (AFM) to probe the physical properties of the LPS of P. aeruginosa strain PA103 (serogroup O11) in situ. This strain contains a mixture of long and very long polymers of O antigen, regulated by two different genes. For this analysis, we studied the wild-type strain and four mutants, ΔWzz1 (producing only very long LPS), ΔWzz2 (producing only long LPS), DΔM (with both the wzz1 and wzz2 genes deleted), and Wzy::GM (producing an LPS core oligosaccharide plus one unit of O antigen). Forces of adhesion between the LPS on these strains and the silicon nitride AFM tip were measured, and the Alexander and de Gennes model of steric repulsion between a flat surface and a polymer brush was used to calculate the LPS layer thickness (which we refer to as length), compressibility, and spacing between the individual molecules. LPS chains were longest for the wild-type strain and ΔWzz1, at 170.6 and 212.4 nm, respectively, and these values were not statistically significantly different from one another. Wzy::GM and DΔM have reduced LPS lengths, at 34.6 and 37.7 nm, respectively. Adhesion forces were not correlated with LPS length, but a relationship between adhesion force and bacterial pathogenicity was found in a mouse acute pneumonia model of infection. The adhesion forces with the AFM probe were lower for strains with LPS mutations, suggesting that the wild-type strain is optimized for maximal adhesion. Our research contributes to further understanding of the role of LPS in the adhesion and virulence of P. aeruginosa.


Body weight, weight perceptions and food intake patterns. A cross-sectional study among male recruits in the Norwegian National Guard.
Young men tend to have a low intake of vegetables and fruit. Unfortunately, this group is difficult to reach with health information. Furthermore, knowledge about weight perceptions and the relationship to food behaviour among young men is scant. The purpose of this study was to explore the relationship between BMI, health and weight perceptions and food intake patterns among young men in the military. Data were collected with a 4-day food diary among 578 male recruits (age 18-26, mean 19.7) in the Norwegian National Guard (response rate 78%), in addition to a questionnaire, including questions about health and weight perceptions, and food frequency when still living at home. Weight and height were objectively measured. Food patterns were explored with principal component analysis, based on the diary data. A multivariate linear regression analysis determined the association between BMI and food patterns, and attitudes to health and slenderness, adjusting for smoking, physical activity and phase of data collection. Twenty eight percent of the recruits were overweight/obese (BMI > 25 kg/m(2)). Two-thirds meant that it is important for them to be slender, and these recruits reported more of both light (p = 0.025) and hard (p = 0.016) physical activity than the others. It was a positive association between the recruits' food frequency at home, and the amount of intake in the military camp for several food items. A principal component analysis identified three distinct food patterns, loading on 1) plant foods, 2) fast food/soft drinks, 3) milk/cereals. Those who stated that it is important for them to be slender, or to have good health, did not have significantly different food intake patterns than the others. BMI was inversely related to scores on the plant food pattern, and positive attitudes to slenderness. The majority of the recruits find it important to be slender. This orientation had a bearing on their physical activity pattern, but less on the food intake pattern. The data also indicate that subjects with high intakes of plant foods were less likely to have a high BMI than others. It is important to raise awareness of healthy eating in young men.


Increasing the molecular contacts between maurotoxin and Kv1.2 channel augments ligand affinity.
Scorpion toxins interact with their target ion channels through multiple molecular contacts. Because a "gain of function" approach has never been described to evaluate the importance of the molecular contacts in defining toxin affinity, we experimentally examined whether increasing the molecular contacts between a toxin and an ion channel directly impacts toxin affinity. For this purpose, we focused on two scorpion peptides, the well-characterized maurotoxin with its variant Pi1-like disulfide bridging (MTX(Pi1)), used as a molecular template, and butantoxin (BuTX), used as an N-terminal domain provider. BuTX is found to be 60-fold less potent than MTX(Pi1) in blocking Kv1.2 (IC(50) values of 165 nM for BuTX versus 2.8 nM for MTX(Pi1)). Removal of its N-terminal domain (nine residues) further decreases BuTX affinity for Kv1.2 by 5.6-fold, which is in agreement with docking simulation data showing the importance of this domain in BuTX-Kv1.2 interaction. Transfer of the BuTX N-terminal domain to MTX(Pi1) results in a chimera with five disulfide bridges (BuTX-MTX(Pi1)) that exhibits 22-fold greater affinity for Kv1.2 than MTX(Pi1) itself, in spite of the lower affinity of BuTX as compared to MTX(Pi1). Docking experiments performed with the 3-D structure of BuTX-MTX(Pi1) in solution, as solved by (1)H-NMR, reveal that the N-terminal domain of BuTX participates in the increased affinity for Kv1.2 through additional molecular contacts. Altogether, the data indicate that acting on molecular contacts between a toxin and a channel is an efficient strategy to modulate toxin affinity.


Designer DNA-silica/carbon nanotube nanocomposites for traceable and targeted drug delivery.
Due to their unique properties like porosity, high water content, softness and biocompatibility, hydrogels are of great interest for biomedical applications such as tissue engineering and drug delivery. We describe a programmable drug delivery system that is based on highly biocompatible SiNP/CNT-DNA nanocomposites, which can be synthesized in a highly modular fashion from DNA-functionalized carbon nanotubes and silica nanoparticles via enzymatic rolling circle amplification. Specific molecular recognition properties were implemented into the materials by DNA sequence design, as demonstrated by incorporation of GC/CG-rich stem loop and aptamer motifs that enable selective binding of intercalating drugs and cell surface receptors, respectively. In a proof-of-concept study we demonstrate the utility of this approach by targeting nanocomposites loaded with the anthracycline drug doxorubicin to HeLa cancer cells. Our observation that these designer materials work more efficiently than the pure drug alone suggests that further developments of the concept might be useful to selectively trigger more complex cellular pathways.


Biosafety: guidelines for working with pathogenic and infectious microorganisms.
This unit, in conjunction with local and national guidelines and regulations, provides the basic biosafety information needed to perform the procedures detailed in this manual. Topics discussed include routine precautions when working with biohazards, disinfectants, disposal of biohazards, biosafety levels (as established by the U.S. National Institutes of Health, and the Centers for Disease Control and Prevention), animal facilities, and clinical laboratories. In addition, resources for more information are provided in the Literature Cited and Key References sections and in URLs given text and within the Internet Resources section.


Unicompartmental knee arthroplasty survivorship is lower than TKA survivorship: a 27-year Finnish registry study.
Balancing the relative advantages and disadvantages of unicompartmental knee arthroplasties (UKAs) against those for TKAs can be challenging. Survivorship is one important end point; arthroplasty registers repeatedly report inferior midterm survival rates, but longer-term data are sparse. Comparing survival directly by using arthroplasty register survival reports also may be inadequate because of differences in indications, implant designs, and patient demographics in patients having UKAs and TKAs. The aims of this study were to assess the survivorship of UKA in the context of one large, northern European registry, and to compare the rates of survivorship with those of cemented TKAs performed for primary knee osteoarthritis during the same 27-year period. From the Finnish Arthroplasty Register, we obtained the data for 4713 patients undergoing UKAs for primary osteoarthritis (mean age, 63.5 years; minimum followup, 0 years; mean, 6.0 years; range, 0-24 years) who had surgical revision between 1985 and 2011. From this cohort, we calculated the Kaplan-Meier survivorship for revision performed for any reason and compared it with the survivorship of 83,511 patients (mean age, 69.5 years; minimum followup 0 years; mean, 6.4 years; range, 0-27 years) with TKAs treated for primary osteoarthritis during the same period. Data were adjusted for age and sex in a comparative analysis. Kaplan-Meier survivorship of UKAs was 89.4% at 5 years, 80.6% at 10 years, and 69.6% at 15 years; the corresponding rates for TKAs were 96.3%, 93.3%, and 88.7%, respectively. UKAs had inferior long-term survivorship compared with cemented TKAs, even after adjusting for the age and sex of the patients (hazard ratio 2.2, p < 0.001). A UKA offers tempting advantages compared with a TKA; however, the revision frequency for UKAs in widespread use, as measured in a large, national registry, was poorer than that of TKAs. When choosing between a UKA and a TKA, patients should be informed of advantages of both procedures, but they also should be advised about the generally higher revision risk after UKA. Level III, therapeutic study. See the Instructions for Authors for a complete description of levels of evidence.


Measurement of regional neuronal removal of norepinephrine in man.
We describe here and validate an in vivo technique to measure the regional proportionate removal of norepinephrine (NE) by neuronal uptake (Uptake1) in man. The measurement is based on the steady-state arterial and venous concentrations of tritiated NE and tritiated isoproterenol (ISO) during simultaneous infusion of both. The validity of this technique depends on the removal of circulating NE, but not of ISO, by sympathetic nerve endings and on there being no other factor contributing to the net difference in the plasma disappearance of these catecholamines. To test these hypotheses, we compared the removal of NE in the arm with that of ISO in 14 people and the effects of pretreatment with the specific inhibitor of Uptake1, desipramine, in 8 people. In all the subjects a greater percent of NE than of ISO was removed during passage of blood through the forearm (54 vs. 46%, P less than 0.0001). Pretreatment with desipramine decreased significantly the removal of NE to virtually exactly that of ISO. The difference in NE and ISO clearances by arm tissues was therefore completely accounted for by Uptake1. About 15% of infused NE which is removed in the arm is removed by Uptake1. The ability to measure regional Uptake1 should contribute to better understanding of the relationship between circulating levels of plasma NE and sympathetic neural activity and may allow detection of abnormalities of neuronal norepinephrine removal in clinical disease states.


Cutaneous stigmata frequently found in the diabetic limb.
Dr. Brenner provides us with a look at some of the common skin problems seen in patients who have diabetes mellitus. As he points out, several of these skin conditions precede the actual clinical and laboratory diagnosis of diabetes by several months to several years. Podiatrists may well be the first physicians to see these prodromal dermopathies and be able to make an early diagnosis and aid the patients in obtaining early treatment.


Site-specific mutagenesis method which completely excludes wild-type DNA from the transformants.
A highly efficient site-specific mutagenesis method has been devised to exclude wild-type DNA from incorporation into the transformed cells. Two complementary oligonucleotides, corresponding to a target sequence of a DNA molecule and containing an insertion mutation which created an endonuclease restriction site, were synthesized. By using the wild-type DNA molecule flanked by two restriction sites on each side of the target region as a template, the two oligonucleotide primers were extended, enriched, and isolated. The extended products, in turn, were used as templates in a polymerase chain reaction to obtain a mutagenized double-stranded DNA fragment which was conveniently cloned into plasmids by using the flanking restriction sites. Escherichia coli cells transformed by these plasmids were subject to large-scale analysis. One hundred percent of the transformants examined by colony hybridization, restriction enzyme analysis, and DNA sequencing were found to contain the mutant DNA sequence.


Radiation-induced leiomyosarcoma of the maxillofacial region: facial reconstruction with implant-retained prosthesis.
Radiation-induced sarcoma represents an exceptional rarity in the head and neck region. The treatment is surgical excision with negative margins and follow-up. After surgery, reconstruction of the defects can be accomplished either surgically or prosthetically. Implant-retained maxillofacial prostheses may offer an acceptable solution. In this report, a large facial defect resulting from radiation-induced leiomyosarcoma was reconstructed with implant-retained prosthesis. A patient with a large facial defect resulting from tumor surgery presented for reconstructive treatment. The primary tumor was fibrous histiosarcoma. Four years after the patient was treated with surgical resection and subsequent irradiation, leiomyosarcoma was diagnosed in the defect site. Leiomyosarcoma was treated with complete surgical resection, and the defect was reconstructed with implant-retained prosthesis. The patient was evaluated every 6 months. Implants provided secure retention to the prosthesis. The patient was satisfied with the appearance of the prosthesis. At 12 months' examination, functioning implants were successful; however, color fade of the prosthesis was observed. Radiation-induced leiomyosarcoma, which is an uncommon tumor in the head and neck region, was treated by means of large surgical resection. Implant-retained prosthetic rehabilitation of the patient resulted in acceptable functional and cosmetic results, and the patient has resumed social activities.


A dynamic channel assignment policy through Q-learning.
One of the fundamental issues in the operation of a mobile communication system is the assignment of channels to cells and to calls. Since the number of channels allocated to a mobile communication system is limited, efficient utilization of these communication channels by using efficient channel assignment strategies is not only desirable but also imperative. This paper presents a novel approach to solving the dynamic channel assignment (DCA) problem by using a form of realtime reinforcement learning known as Q-learning in conjunction with neural network representation. Instead of relying on a known teacher, the system is designed to learn an optimal channel assignment policy by directly interacting with the mobile communication environment. The performance of the Q-learning-based DCA was examined by extensive simulation studies on a 49-cell mobile communication system under various conditions. Comparative studies with the fixed channel assignment (FCA) scheme and one of the best dynamic channel assignment strategies, MAXAVAIL, have revealed that the proposed approach is able to perform better than the FCA in various situations and capable of achieving a performance similar to that achieved by the MAXIAVIAL, but with a significantly reduced computational complexity.


Cadmium accumulation in different pakchoi cultivars and screening for pollution-safe cultivars.
The selection and breeding of pollution-safe cultivars (PSCs) is a practicable and cost-effective approach to minimize the influx of heavy metal to the human food chain. In this study, both pot-culture and field experiments were conducted to identify and screen out cadmium pollution-safe cultivars (Cd-PSCs) from 50 pakchoi (Brassica rapa L. ssp. chinensis) cultivars for food safety. When treated with 1.0 or 2.5 mg/kg Cd, most of the pakchoi cultivars (>70%) showed greater or similar shoot biomass when compared with the control. This result indicates that pakchoi has a considerable tolerance to soil Cd stress. Cd concentrations in the shoot varied significantly (P<0.05) between cultivars: in two Cd treatments (1.0 and 2.5 mg/kg), the average values were 0.074 and 0.175 mg/kg fresh weight (FW), respectively. Cd concentrations in the shoots of 14 pakchoi cultivars were lower than 0.05 mg/kg FW. In pot-culture experiments, both enrichment factors (EFs) and translocation factors (TFs) of six pakchoi cultivars were lower than 1.0. The field studies further confirmed that the Hangzhouyoudonger, Aijiaoheiye 333, and Zaoshenghuajing cultivars are Cd-PSCs, and are therefore suitable for growth in low Cd-contaminated soils (≤1.2 mg/kg) without any risk to food safety.


Targeted cell entry of lentiviral vectors.
Retargeting of lentiviral vector entry to cell types of interest is a key factor in improving the safety and efficacy of gene transfer. In this study we show that the retargetable envelope glycoproteins of measles virus (MV), namely, the hemagglutinin (H) responsible for receptor recognition and the fusion protein (F), can pseudotype human immunodeficiency virus 1 (HIV-1) vectors when their cytoplasmic tails are truncated. We then pseudotyped HIV-1 vectors with MV glycoproteins displaying on H either the epidermal growth factor or a single-chain antibody directed against CD20, but without the ability to recognize their native receptors. Gene transfer into cells that expressed the targeted receptor was several orders of magnitude more efficient than into cells that did not. High-target versus nontarget cell discrimination was demonstrated in mixed cell populations, where the targeting vector selectively eliminated CD20-positive cells after suicide gene transfer. Remarkably, primary human CD20-positive B lymphocytes were transduced more efficiently by the CD20-targeted vector than by a vector pseudotyped with the vesicular stomatitis virus G (VSV-G) protein. In addition, the CD20-targeted vector was able to transduce even unstimulated primary B cells, whereas VSV-G pseudotyped vectors were unable to do so. Because MV enters cells through direct fusion at the cell membrane, this novel targeting system should be widely applicable.


Bicarbonate buffered ultrafiltration leads to a physiologic priming solution in pediatric cardiac surgery.
Pediatric cardiopulmonary bypass (CPB) involves a high ratio of priming blood volume to patient blood volume. The composition of packed red blood cells (RBCs) is very unphysiological in terms of acid-base, electrolyte and metabolite values. Therefore, we tested the hypothesis whether ultrafiltration of the prime and replacement with bicarbonate buffered hemofiltration solution (BB-HS) is sufficient for reducing the metabolic load and reaching a physiologic state. For planned surgery of congenital heart defects with cardiopulmonary bypass, 20 CPB circuits were primed with BB-HS, gelatin and 1 unit of RBCs. The fluid was hemofiltrated using an ultrahemofilter at 300 ml/min until approximately 1000 ml of ultrafiltrate was restored with BB-HS. Blood gas analyses were obtained from the priming blood, once before and once after bicarbonate buffered ultrafiltration (BBUF). The measured substrates decreased significantly (P<0.001) after BBUF (glucose from 13.0+/-2.6 to 6.3+/-1.0 and lactate from 3.8+/-1.5 to 2.3+/-1.0 mmol/l). Acid-base parameters increased (P<0.001) to normal or high normal values (pH from 7.01+/-0.09 to 7.68+/-0.12; HCO(3) from 12.1+/-2.4 to 25.4+/-3.6 mmol/l and BE from -15.4+/-3.6 to -0.8+/-3.7 mmol/l). Even the electrolytes sodium, potassium and calcium changed significantly (P<0.001) toward the physiologic range. BBUF is an efficient method of reducing the metabolic load of priming. After BBUF, even the electrolyte and acid-base balance reached a physiologic state, which is important for minimizing electrolyte and acid-base disturbances after initiation of CPB.


In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae.
The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.


Predictive effects of urinary liver-type fatty acid-binding protein for deteriorating renal function and incidence of cardiovascular disease in type 2 diabetic patients without advanced nephropathy.
To improve prognosis, it is important to predict the incidence of renal failure and cardiovascular disease in type 2 diabetic patients before the progression to advanced nephropathy. We investigated the predictive effects of urinary liver-type fatty acid-binding protein (L-FABP), which is associated with renal tubulointerstitial damage, in renal and cardiovascular prognosis. Japanese type 2 diabetic patients (n = 618) with serum creatinine ≤1.0 mg/dL and without overt proteinuria were enrolled between 1996 and 2000 and followed up until 2011. Baseline urinary L-FABP was measured with an enzyme-linked immunosorbent assay. The primary end points were renal and cardiovascular composites (hemodialysis, myocardial infarction, angina pectoris, stroke, cerebral hemorrhage, and peripheral vascular disease). The secondary renal outcomes were the incidence of a 50% decline in estimated glomerular filtration rate (eGFR), progression to an eGFR <30 mL/min/1.73 m(2), and the annual decline rate in eGFR. During a 12-year median follow-up, 103 primary end points occurred. The incidence rate of the primary end point increased in a stepwise manner with increases in urinary L-FABP. In Cox proportional hazards analysis, the adjusted hazard ratio in patients with the highest tertile of urinary L-FBAP was 1.93 (95% CI 1.13-3.29). This relationship was observed even when analyzed separately in normoalbuminuria and microalbuminuria. Patients with the highest tertile of urinary L-FABP also demonstrated a higher incidence of the secondary renal outcomes. Our results indicate that urinary L-FABP may be a predictive marker for renal and cardiovascular prognosis in type 2 diabetic patients without advanced nephropathy.


Stabilizing ruthenium dioxide with cation-anchored sulfate for durable oxygen evolution in proton-exchange membrane water electrolyzers.
Ruthenium dioxide is the most promising alternative to the prevailing but expensive iridium-based catalysts for the oxygen evolution reaction in proton-exchange membrane water electrolyzers. However, the under-coordinated lattice oxygen of ruthenium dioxide is prone to over-oxidation, and oxygen vacancies are formed at high oxidation potentials under acidic corrosive conditions. Consequently, ruthenium atoms adjacent to oxygen vacancies are oxidized into soluble high-valence derivatives, causing the collapse of the ruthenium dioxide crystal structure and leading to its poor stability. Here, we report an oxyanion protection strategy to prevent the formation of oxygen vacancies on the ruthenium dioxide surface by forming coordination-saturated lattice oxygen. Combining density functional theory calculations, electrochemical measurements, and a suite of operando spectroscopies, we showcase that barium-anchored sulfate can greatly impede ruthenium loss and extend the lifetime of ruthenium-based catalysts during acidic oxygen evolution, while maintaining the activity. This work paves a new way for designing stable and active anode catalysts toward acidic water splitting.


At the limits of bisphosphonio-substituted stannylenes.
Donor stabilization of Sn(II) and Pb(II) halides with 1,1'-ferrocenylene bridged bisphosphanes has been explored for Fe(C5H4P(C6H5)2)2 (dppf), and Fe(C5H4PH(C4H9))2. These bisphosphanes are reacted with SnBr2 and PbCl2 with and without additional Lewis acid (AlCl3) forming acyclic and cyclic donor adducts from which the latter represent bisphosphoniotetrylenes. Since dynamic exchange in solution is observed, characterization includes solution and solid-state NMR in addition to SC-XRD, amended by DFT calculations.


Intraregional differences in renal function in the Northern Netherlands: The Lifelines Cohort Study.
Although the interregional disparity in chronic kidney disease (CKD) prevalence has been reported globally, little is known about differences in CKD prevalence within a region. We aimed to study the intraregional distribution of renal function in the Northern Netherlands and identify determinants of geographical differences in renal function. We included 143,735 participants from the Lifelines population-based cohort in the Northern Netherlands. Spatial analysis was performed to identify regional clusters of lower eGFR (cold spots) and higher eGFR (hot spots) at the postal code level, without and with adjustment for clinical risk factors. Multivariate logistic regression was used to identify the contribution of neighborhood-level health-related behaviors, socioeconomic status, and environmental factors (air pollution parameters, urbanity) to regional clustering of lower eGFR. Significant spatial clustering of renal function was found for eGFR as well as for early stage renal function impairment (eGFR<90 ml/min/1.73 m2), (p<0.001). Spatial clustering persisted after adjustment of eGFR for clinical risk factors. In adjusted cold spots, the aggregate eGFR was lower (mean ± SD: 96.5±4.8 vs. 98.5±4.0 ml/min/1.73 m2, p = 0.001), and the prevalence of early stage renal function impairment (35.8±10.9 vs. 28.7±9.8%, p<0.001) and CKD stages 3-5 was higher (median (interquartile range): 1.2(0.1-2.4) vs 0(0-1.4)%, p<0.001) than in hot spots. In multivariable logistic regression, exposure to NO2 (Odd ratio [OR], 1.45; 95% confidence interval [95% CI], 1.19 to 1.75, p<0.001) was associated with cold spots (lower renal function), whereas proportion of fat intake in the diet (OR, 0.68; 95%CI, 0.48-0.97, P = 0.031) and income (OR, 0.91; 95%CI, 0.86-0.96, p<0.001) for median level income) were inversely related. Significant intraregional clustering of renal function, early renal function impairment and CKD were observed in the Northern Netherlands even after adjustment for renal function-related clinical risk factors. Environmental (air pollution), neighborhood-level socioeconomic factors and diet are determinants of intraregional renal function distribution. Spatial analysis might be a useful adjunct to guide public health strategies for the prevention of CKD.


Healing of two-wall intra-bony defects treated with a novel EMD-liquid-A pre-clinical study in monkeys.
To investigate the effect of a novel enamel matrix derivative formulation (EMD-liquid or Osteogain) combined with an absorbable collagen sponge (ACS) on periodontal wound healing in intra-bony defects in monkeys. Chronic two-wall intra-bony defects were created at the distal aspect of eight teeth in three monkeys (Macaca fascicularis). The 24 defects were randomly assigned to one of the following treatments: (i) open flap debridement (OFD) + ACS alone, (ii) OFD + Emdogain + ACS (Emdogain/ACS), (iii) OFD + Osteogain + ACS (Osteogain/ACS) or (iv) OFD alone. At 4 months, the animals were euthanized for histologic evaluation. Osteogain/ACS resulted in more consistent formation of cementum, periodontal ligament and bone with limited epithelial proliferation compared to OFD alone, Emdogain/ACS and OFD + ACS. Among the four treatment groups, the Osteogain/ACS group demonstrated the highest amount of regenerated tissues. However, complete periodontal regeneration was not observed in any of the defects in the four groups. The present findings indicate that in two-wall intra-bony defects, reconstructive surgery with Osteogain/ACS appears to be a promising novel approach for facilitating periodontal wound healing/regeneration, thus warranting further clinical testing.


Uniform motion blur in Poissonian noise: blur/noise tradeoff.
In this paper we consider the restoration of images corrupted by both uniform motion blur and Poissonian noise. We formulate an image formation model that explicitly takes into account the length of the blur point-spread function and the noise level as functions of the exposure time. Further, we present an analysis of the achievable restoration performance by showing how the root mean squared error varies with respect to the exposure time. It turns out that the worst situations are represented by either too short or too long exposure times. In between there exists an optimal exposure time that maximizes the restoration performance, balancing the amount of blur and noise in the observation. We justify such result through a mathematical analysis of the signal-to-noise ratio in Fourier domain; this study is then validated by deblurring synthetic data as well as camera raw data.


Tympanomastoidectomy versus parenteral antibiotic therapy for pediatric otorrhea.
To evaluate the efficacy of tympanomastoidectomy versus parenteral antibiotic therapy for otorrhea as a result of chronic suppurative otitis media (CSOM) without cholesteatoma in the pediatric population. A retrospective review of 221 patients treated for otorrhea at a tertiary academic pediatric hospital was performed to evaluate the impact of tympanomastoidectomy versus parenteral antibiotic therapy on resolution of otorrhea. Inclusion criteria were age 0-18 years, prior treatment with otic and/or oral antibiotic, prior history of tympanostomy tube placement for recurrent otitis media, history of otorrhea, treatment with tympanomastoidectomy or parenteral antibiotic therapy, and follow-up of at least 1 month after intervention. Time to resolution was compared between the two modalities adjusting for age, bilateral ear disease status, and comorbidities using a Cox proportional hazard model. Eighty-three ears from 58 children met the inclusion criteria. Ears that initially underwent tympanomastoidectomy had a significantly shorter time to resolution of symptoms (median time to resolution) 9 months (95 % confidence interval CI: 6.2-14.8) vs. 48.5 months (95 % lower CI 9.4, p = 0.006). On multivariate analysis, however, only bilateral ear disease status was independently associated with time to resolution of symptoms (hazard ratio 0.4, 95 % CI 0.2-0.9, p = 0.03). There was no statistically significant difference in the rate of treatment-related complications when comparing tympanomastoidectomy to parenteral antibiotic therapy (p = 0.37). When adjusting for age, bilateral ear disease status, and comorbidities, there does not appear to be a significant difference in time to resolution of symptoms when comparing parenteral antibiotic therapy to tympanomastoidectomy. An informed discussion regarding risks and benefits of each approach should be employed when deciding on the next step in management for patients with CSOM who have failed more conservative therapies.


Expression of the zinc finger gene fez-like in zebrafish forebrain.
Anterior-posterior (A-P) patterning in the neuroectoderm is established during gastrulation in zebrafish and amphibians. We isolated a novel zinc-finger gene fez-like (fezl) from zebrafish, which displays sequence similarities to Xenopus Fez. The fezl transcripts were detected in the anterior edge of neuroectoderm, the prospective dorsal forebrain, from the late gastrula (80% epiboly stage) to the mid-segmentation period. fezl was also expressed in the ventral forebrain overlying the prechordal plate at these stages. The expression of fezl was enhanced in embryos expressing the Wnt inhibitor Dkk1 and reduced in embryos expressing Wnt8b. The expression in the ventral forebrain was eliminated in the one-eyed pinhead mutant and the antivin RNA-injected embryos, which lack the prechordal plate. Radiation hybrid mapping revealed that the fezl gene is localized to linkage group 11.


No association between major depression with and without childhood adversity and the stress hormone copeptin.
Background: Adverse childhood experiences (ACE) are associated with an increased risk of major depressive disorder (MDD) and hypothalamic-pituitary-adrenal (HPA) axis dysregulation. Within the HPA axis, corticotropin-releasing hormone and vasopressin (AVP) synergistically stimulate the release of adrenocorticotropic hormone, which promotes cortisol release. The cleavage product copeptin is produced during AVP synthesis and is a surrogate marker of AVP release. Children with ACE and young adults with depressive symptoms have higher levels of copeptin than healthy controls. Objective: To uncover the effects of MDD and ACE on copeptin levels in adult females. Methods: We recruited 94 women (mean age: 34.0 ± 3.6 years): 23 with MDD and ACE, 24 with MDD without ACE, 22 with ACE without MDD, and 25 healthy controls. ACE was defined as repeated sexual or physical abuse at least once a month over at least one year before the age of 18 years. MDD was defined by the DSM-IV criteria. Copeptin plasma levels were measured with an immunoluminometric assay. Results: The four groups did not differ in demographic variables. We found a significant negative correlation between body mass index (BMI) and copeptin plasma levels (r = -.21; p = .045). Copeptin plasma levels did not differ between the four groups after controlling for BMI. Conclusion: Neither MDD nor ACE was associated with altered plasma copeptin levels. Thus, copeptin does not seem to play a major role in MDD and ACE in adult females.


A critical synthesis of the role of the pharmacist in oral healthcare and management of medication-related osteonecrosis of the jaw.
To consolidate extant published evidence in relation to the potential of integrating oral healthcare for patients at risk of developing medication-related osteonecrosis of the jaw (MRONJ). A critical synthesis and consolidation of five publications was undertaken. As a mechanism of situating the extant work within the context of primary healthcare provision, the Rainbow Model of Integrated Care was applied as a theoretical lens through which the conceptual findings could be collectively applied to practice. The critical synthesis revealed a thematic emergence relating to both formative and normative integration. The most salient of these were the identification of limited shared clinical records, and disconnection of oral healthcare provision from patients' general medical care. The three levels of the Rainbow Model of Integrated Care reflected a series of issues for address. In the context of collaborative, multi-disciplinary working for patients at risk of development of MRONJ, pharmacists are a professional group which this research reveals to be an underutilised resource. Reduction of oral health inequality at all levels of patient care is a key priority and this research highlights areas for address in relation to requirements for interprofessional education, optimal communication and policies reflective and facilitative of these.


Influence of rate of change of frequency on the overall pitch of frequency-modulated tones.
The mechanism(s) determining pitch may assign less weight to portions of a sound where the frequency is changing rapidly. The present experiments explored the possible effect of this on the overall pitch of frequency-modulated sounds. Pitch matches were obtained between an adjustable unmodulated sinusoid and a sinusoidal carrier that was frequency modulated using a highly asymmetric function with the form of a repeating U or inverted U shaped function. The amplitude was constant during the 400-ms presentation time of each stimulus, except for 10-ms raised-cosine onset and offset ramps. In experiment 1, the carrier level was 50 dB SPL and the geometric mean of the instantaneous frequency of the modulated carrier, fc, was either 0.5, 1, 2, or 8 kHz. The modulation rate (fm) was 5, 10, or 20 Hz. The overall depth (maximum to minimum) of the FM was 8% of fc. For all carrier frequencies, the matched frequency was shifted away from the mean carrier frequency, downwards for the U shaped function stimuli and upwards for the repeated inverted U shaped function stimuli. The shift was typically slightly greater than 1% of fc, and did not vary markedly with fc. The effect of fm was small, but there was a trend for the shifts to decrease with increasing fm for fc = 0.5 kHz and to increase with increasing fm for fc = 2 kHz. In experiment 2, the carrier level was reduced to 20 dB SL and matches were obtained only for fc = 2 kHz. Shifts in matched frequency of about 1% were still observed, but the trend for the shifts to increase with increasing fm no longer occurred. In experiment 3, matches were obtained for a 4-kHz carrier at 50 dB SPL. Shifts of about 1% again occurred, which did not vary markedly with fm. The shifts in matched frequency observed in all three experiments are not predicted by models based on the amplitude- or intensity-weighted average of instantaneous frequency (EWAIF or IWAIF). The shifts (and the pitch shifts observed earlier for two-tone complexes and for stimuli with simultaneous AM and FM) are consistent with a model based on the assumption that the overall pitch of a frequency-modulated sound is determined from a weighted average of period estimates, with the weight attached to a given estimate being inversely related to the short-term rate of change of period and directly related to a compressive function of the amplitude.


Seasonally increasing parasite load is associated with microbiome dysbiosis in wild bumblebees.
The microbiome is increasingly recognized for its complex relationship with host fitness. Bumblebees are host to a characteristic gut microbiome community that is derived and reinforced through social contact between individuals. The bumblebee microbiome is species-poor, and primarily composed from a small number of core taxa that are associated with the greater tribe of corbiculate bees. Experimental findings support a role for the core bumblebee microbiome in resistance to severe infections by a common trypanosomal parasite, Crithidia bombi . However, most studies have been small in scale, often considering just one or two bumblebee species, or making use of commercially-reared bees. To better understand the microbiome diversity of wild populations, we have deeply sampled field populations of ten sympatric species found throughout central and down east Maine in a three-year microbiome field survey. We have used 16S amplicon sequencing to produce microbiome community profiles, and qPCR to screen samples for infections by Crithidia bombi . The breadth of our dataset has enabled us to test for seasonal and interspecific trends in the microbiome community. Controlling for these external sources of variation, we have identified microbial factors associated with infection and parasite load that support the role of the core microbiome in resistance to severe infection.