Genetic structure and evolution of natural populations of viruses causing the tomato yellow leaf curl disease in Spain.
The population structure and genetic variation of two begomoviruses: tomato yellow leaf curl Sardinia virus (TYLCSV) and tomato yellow leaf curl virus (TYLCV) in tomato crops of Spain were studied from 1997 until 2001. Restriction digestion of a genomic region comprised of the CP coat protein gene (CPR) of 358 TYLC virus isolates enabled us to classify them into 14 haplotypes. Nucleotide sequences of two genomic regions: CPR, and the surrounding intergenic region (SIR) were determined for at least two isolates per haplotype. SIR was more variable than CPR and showed multiple recombination events whereas no recombination was detected within CPR. In all geographic regions except Murcia, the population was, or evolved to be composed of one predominant haplotype with a low genetic diversity (<0.0180). In Murcia, two successive changes of the predominant haplotype were observed in the best studied population. Phylogenetic analysis showed that the TYLCSV sequences determined clustered with sequences obtained from the GenBank of other TYLCSV Spanish isolates which were clearly separated from TYLCSV Italian isolates. Most of our TYLCV sequences were similar to those of isolates from Japan and Portugal, and the sequences obtained from TYLCV isolates from the Canary island of Lanzarote were similar to those of Caribbean TYLCV isolates.


Review of spinal epidural cavernous hemangioma.
The characteristics of spinal epidural cavernous hemangioma without primary origin in the vertebral bone were evaluated in 54 patients including our new case. The 36 male and 18 female patients were aged 5 to 78 years (mean 47 years). Most lesions were in the thoracic spine (80%) and on the dorsal side of the spinal cord (93%). The clinical course was mostly slowly progressive, with myelopathy in 33% at onset and 83% at admission. The lesion appeared isointense to the spinal cord on T1-weighted imaging, and isointense or slightly hypointense to the cerebrospinal fluid on T2-weighted imaging. Lesion without hemorrhage showed prominent homogeneous enhancement after administration of gadolinium-diethylenetriaminepenta-acetic acid because of the sinusoidal channel structure. Heterogeneous enhancement was caused by hematoma and/or post-hemorrhagic degeneration. The differential diagnosis of this disease includes metastatic tumor, Ewing's sarcoma, chordoma, eosinophilic granuloma, sarcoidosis, lipoma, hypertrophy of the posterior longitudinal ligament or the ligamentum flavum, meningioma, and neurinoma. The relationships between clinical course and surgery or outcome suggest that early diagnosis and total removal of the lesion before massive lesional bleeding occurs are necessary for a good outcome.


Population level inference for multivariate MEG analysis.
Multivariate analysis is a very general and powerful technique for analysing Magnetoencephalography (MEG) data. An outstanding problem however is how to make inferences that are consistent over a group of subjects as to whether there are condition-specific differences in data features, and what are those features that maximise these differences. Here we propose a solution based on Canonical Variates Analysis (CVA) model scoring at the subject level and random effects Bayesian model selection at the group level. We apply this approach to beamformer reconstructed MEG data in source space. CVA estimates those multivariate patterns of activation that correlate most highly with the experimental design; the order of a CVA model is then determined by the number of significant canonical vectors. Random effects Bayesian model comparison then provides machinery for inferring the optimal order over the group of subjects. Absence of a multivariate dependence is indicated by the null model being the most likely. This approach can also be applied to CVA models with a fixed number of canonical vectors but supplied with different feature sets. We illustrate the method by identifying feature sets based on variable-dimension MEG power spectra in the primary visual cortex and fusiform gyrus that are maximally discriminative of data epochs before versus after visual stimulation.


Safety and efficacy of Avatec® 150G (lasalocid A sodium) as a feed additive for chickens for fattening and chickens reared for laying.
Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of Avatec® 150G (lasalocid A sodium) for chickens for fattening and chickens reared for laying. In 2017, the FEEDAP Panel was not able to conclude on a safe dose for chickens for fattening and chickens reared for laying and no conclusion could be drawn on the anticoccidial efficacy of the additive at the lowest proposed used level (75 mg lasalocid A sodium/kg feed) in these species. In the present assessment, the applicant submitted new tolerance and efficacy studies in chickens for fattening to address the concerns identified by the FEEDAP Panel in its former opinion. In addition, the applicant proposed to decrease the maximum of the dose range from 125 to 100 mg lasalocid A sodium/kg complete feed. Based on the additional information, the FEEDAP Panel concludes that no safe level of lasalocid A sodium from Avatec® 150G in feed for chickens for fattening can be identified. The FEEDAP Panel is not in the position to conclude on the coccidiostatic efficacy of Avatec® 150G for chickens for fattening at the lowest proposed dose level of 75 mg lasalocid A sodium/kg complete feed due to the insufficient number of studies with positive results. The conclusions are extended to chickens reared for laying.


Respiratory arterial pH and PCO2 oscillations in patients with chronic obstructive airways disease.
1. Arterial pH oscillations have been monitored in vivo in patients with well defined chronic obstructive bronchitis, asthma and clinical emphysema. 2. The patients with clinical emphysema were shown to differ from those with chronic obstructive bronchitis on the basis of a number of clinical and physiological criteria. 3. Patients with asthma showed least attenuation of their pH oscillations as a group, in contrast to emphysematous patients who showed most attenuation. In patients with clinical emphysema the attenuation was relatively homogeneous. The patients with chronic obstructive bronchitis showed the full range from normal oscillations (zero attenuation) to zero (complete attenuation). 4. The amplitude and approximate rate of change of upslope of the PaCO2 oscillations in vivo were calculated, from measured pH oscillation amplitudes, using buffer slope values in vitro from Siggaard Anderson [(1962, 1963) Scandinavian Journal of Clinical and Laboratory Investigation, 14, 598-604; 15, 211-217], then dividing the PaCO2 amplitude by half the respiratory period. 5. Mean arterial PCO2 in vitro showed a very strong correlation with the downslope of the pH oscillation in vivo (calculated as for PaCO2 upslope) in patients without clinical emphysema. This correlation would be expected to some extent, owing to the logarithmic relationship of PaCO2 oscillations to pH oscillations. However, the mean arterial PCO2 also showed a very strong correlation with the upslope of the calculated PaCO2 oscillations, again excluding patients with clinical emphysema.


Can the gateway hypothesis, the common liability model and/or, the route of administration model predict initiation of cannabis use during adolescence? A survival analysis--the TRAILS study.
There is substantial research linking tobacco and alcohol use to subsequent cannabis use, yet the specificity of this relationship is still under debate. The aim of this study was to examine which substance use model--the gateway hypothesis, the common liability (CL) model and/or the route of administration model--best explains the relationship between early onset of tobacco and alcohol use and subsequent cannabis use initiation. We used data from 2,113 (51% female) Dutch adolescents who participated in three consecutive assessment waves (mean age: 11.09, 13.56, and 16.27 years, respectively) of the TRacking Adolescents' Individual Lives Survey study. (Pre)adolescent cannabis, tobacco and alcohol use was assessed using the Youth Self-Report and a TRacking Adolescents' Individual Lives Survey developed questionnaire. We found that, during adolescence, early onset of tobacco use does not pose a significantly higher risk of initiating cannabis use than early onset alcohol use. Therefore, we can rule out the route of administration model. Moreover, we found that adolescents who reported early onset comorbid use of both tobacco and alcohol have a higher likelihood to initiate cannabis use than adolescents who have tried either tobacco or alcohol. The gateway hypothesis is not broad enough to explain this finding. Therefore, the CL model best predicts our findings. Future research on adolescent cannabis initiation should focus on testing the robustness of the CL model. Furthermore, identifying adolescents who use both tobacco and alcohol, before the age of 13, may help to curtail the onset of cannabis use.


Delivering high-resolution landmarks using inkjet micropatterning for spatial monitoring of leaf expansion.
Inkjet micropatterning is a versatile deposition technique with broad applications in numerous fields. However, its application in plant science is largely unexplored. Leaf expansion is one of the most important parameters in the field of plant science and many methods have been developed to examine differential expansion rates of different parts of the leaf lamina. Among them, methods based on the tracking of natural landmarks through digital imaging require a complicated setup in which the leaf must remain fixed and under tension. Furthermore, the resolution is limited to that of the natural landmarks, which are often difficult to find, particularly in young leaves. To study the fine scale expansion dynamics of the leaf lamina using artificial landmarks it is necessary to place small, noninvasive marks on a leaf surface and then recover the location of those marks after a period of time. To monitor leaf expansion in two dimensions, at very fine scales, we used a custom designed inkjet micropatterning system to print a grid composed of c. 0.19 mm2 cells on small developing leaves of ivy (Hedera helix) using 40 μm dots at a spacing of c. 91 μm. The leaves in different growing stages were imaged under magnification to extract the coordinates of the marks which were then used in subsequent computer-assisted leaf expansion analyses. As an example we obtained quantified global and local expansion information and created expansion maps over the entire leaf surface. The results reveal a striking pattern of fine-scale expansion differences over short periods of time. In these experiments, the base of the leaf is a "cold spot" for expansion, while the leaf sinuses are "hot spots" for expansion. We have also measured a strong shading effect on leaf expansion. We discuss the features required to build an inkjet printing apparatus optimized for use in plant science, which will further maximize the range of tissues that can be printed at these scales. To apply inkjet micropatterning to plant studies, we have successfully delivered landmarks on ivy leaf surfaces and achieved high-resolution, two-dimensional monitoring of leaf expansion at different growing stages. The measurement is capable of reliably identifying the fine scale changes during plant growth. As well as delivering landmarks, this technology may be used to deliver microscale targeted biological components such as growth hormones, and possibly be used to pattern sensors directly on the leaves.


Provision of high-protein supplement for patients recovering from hip fracture.
We compared clinical outcomes with a standard (Ensure) or a high-protein (Boost HP) liquid nutritional supplement for older adults recovering from hip fracture surgery in a rehabilitation hospital. This randomized, double-blind, parallel-group study compared the clinical effectiveness of a standard (Ensure) with a high-protein (Boost HP) liquid nutritional supplement among patients (n = 46) 60 y or older who recently underwent surgical repair of a hip fracture. Patients were encouraged to drink at least two 8-oz cans (17.8 g/d protein for Ensure versus 30 g/d protein for Boost HP) per day for 28 d. Study measurements included change in Functional Independence Measure between rehabilitation admission and discharge, length of rehabilitation stay, laboratory measures (i.e., serum albumin, prealbumin, and C-reactive protein), physical activity energy expenditure by 7-d triaxial accelerometry, and dietary intake by three random, telephonic, 24-h dietary recalls. There were no significant group differences with respect to age, sex, acute hospital days, hip fracture assessment parameters, or surgical treatment. Consumption of supplement (260 oz/28 d of Ensure versus 239 oz/28 d of Boost HP) was comparable. There were no differences in complication or adverse event rates during the study. The Boost HP group consumed more protein than the Ensure group (63 versus 50 g, P < 0.048) and had a greater improvement in serum albumin over the 28-d supplementation period (+0.7 versus +0.2 g/dL, P < 0.019). The Boost HP group also consumed more fiber (12 versus 8 g), calcium (821 versus 639 mg), vitamin K (66 versus 45 microg), and phosphorus (1035 versus 833 mg) than did the Ensure group. Rehabilitation length of stay was shorter in the Boost HP than in the Ensure group, although this trend did not reach statistical significance (23 versus 28 d, P = 0.27). Outcome differences were not detected in the Functional Independence Measure. Supplementation was well tolerated in this population and contributed significantly to total dietary intake. Consumption of a high-protein liquid nutritional supplement may offer some benefits by improving visceral protein status.


Urban trails and physical activity: a natural experiment.
The built environment in which a person lives and works is thought to have a strong influence on his or her level of physical activity. However, this belief is largely based on cross-sectional studies underlining the need for prospective studies using natural experiments. This study adopted a quasi-experimental research design with multiple control neighborhoods and was conducted between 2005 and 2007. Data were analyzed in 2008. The subjects were children, adolescents, and adults in free-living conditions within one experimental and two control neighborhoods. An urban greenway/trail was retrofitted in a neighborhood that lacked connectivity of the residential pedestrian infrastructure to nonresidential destinations. The main outcomes were 2-hour counts of directly observed physical activity in the general neighborhood and, at the school level, active transport to school. At the neighborhood level, the 2-hour counts of physical activity significantly increased between 2005 and 2007 (p=0.000) in the intervention neighborhood, with a median increase of 8.0 counts. The control neighborhoods had a significant decrease in counts (p=0.000). The pre- and post-intervention changes between experimental and control neighborhoods were significantly different for total physical activity (p=0.001); walking (p=0.001); and cycling (p=0.038). There was no noted change over time for active transport to school in either the intervention or control neighborhoods. Changes to the pedestrian connectivity of the built environment infrastructure may lead to greater levels of physical activity. However, this positive effect was limited to physical activity at the neighborhood level and not to active transport to school.


A tale of two trails: exploring different paths to success.
This comparative case study investigates 2 successful community trail initiatives, using the Active Living By Design (ALBD) Community Action Model as an analytical framework. The model includes 5 strategies: preparation, promotion, programs, policy, and physical projects. Key stakeholders at 2 sites participated in in-depth interviews (N=14). Data were analyzed for content using Atlas Ti and grouped according to the 5 strategies. PREPARATION: Securing trail resources was challenging, but shared responsibilities facilitated trail development. PROMOTIONS: The initiatives demonstrated minimal physical activity encouragement strategies. Community stakeholders did not coordinate programmatic opportunities for routine physical activity. POLICY: Trails' inclusion in regional greenway master plans contributed to trail funding and development. Policies that were formally institutionalized and enforced led to more consistent trail construction and safer conditions for users. PHYSICAL PROJECTS: Consistent standards for wayfinding signage and design safety features enhanced trail usability and safety. Communities with different levels of government support contributed unique lessons to inform best practices of trail initiatives. This study revealed a disparity between trail development and use-encouragement strategies, which may limit trails' impact on physical activity. The ALBD Community Action Model provided a viable framework to structure cross-disciplinary community trail initiatives.


Na+,K(+)-ATPase activity and hormones in single nephron segments from nephrotic rats.
1. In the nephrotic syndrome the kidneys retain salt and water, which leads to oedema formation. The site of this sodium retention has been localized in the cortical collecting tubule by micropuncture studies. Whether or not this phenomenon is an intrinsic renal problem or is the consequence of changes in hormonal activities is still a matter of discussion. 2. Using the model of puromycin aminonucleoside-induced nephrotic syndrome in the rat, we measured Na+,K(+)-ATPase activity in nephron segments from control and nephrotic rats and investigated the regulatory role of aldosterone and endogenous-ouabain-displacing factor. 3. Nephrotic animals had a twofold increase in Na+,K(+)-ATPase activity in the cortical collecting tubule only (control versus nephrotic: 1065 +/- 68 versus 2081 +/- 274 pmol h-1 mm-1, P = 0.036), which was not modified by adrenalectomy and was independent of the kidney content of endogenous ouabain-displacing factor. Na+,K(+)-ATPase activity in the cortical collecting tubule correlated with the sodium balance in both control and nephrotic rats. 4. The data are consistent with the view that sodium retention in this model of the nephrotic syndrome is a primary event, i.e. an increase in sodium transport throughout the cortical collecting tubule expressed as a twofold increase in Na+,K(+)-ATPase activity which is no longer under hormonal regulation (aldosterone and endogenous ouabain-displacing factor).


Treatment-related myelodysplastic syndrome and acute myelogenous leukemia in patients treated with ibritumomab tiuxetan radioimmunotherapy.
To investigate the incidence of treatment-related myelodysplastic syndrome (t-MDS) and treatment-related acute myelogenous leukemia (t-AML) after treatment with ibritumomab tiuxetan radioimmunotherapy. Analysis of the incidence of t-MDS and t-AML in 746 patients with non-Hodgkin's lymphoma (NHL) treated with the ibritumomab tiuxetan regimen in registration and compassionate-use trials between 1996 and 2002. Nineteen patients (2.5%) developed t-MDS or t-AML at a median follow-up of 4.4 years (range, 0 to 9.3). These malignancies were diagnosed at a median of 5.6 years (range, 1.4 to 13.9) after the diagnosis of NHL and 1.9 years (range, 0.4 to 6.3) after radioimmunotherapy. The annualized rates were 0.3% per year after the diagnosis of NHL and 0.7% per year after treatment. Most patients with t-MDS or t-AML had multiple cytogenetic aberrations, commonly on chromosomes 5 and 7, suggesting an association with previous exposure to chemotherapy. Analysis of data from patients in registration and compassionate-use trials suggests that the annualized incidences of t-MDS and t-AML are consistent with that expected in patients with NHL who have had extensive previous chemotherapy treatment and do not appear to be increased after treatment with the ibritumomab tiuxetan regimen. Cytogenetic testing before treatment with radioimmunotherapy may identify existing chromosomal abnormalities in previously treated patients, particularly those who have been treated with alkylating agents and purine nucleoside analogs and would be at higher risk for t-MDS or t-AML.


Extra-adrenal myelolipoma: report of two cases.
Myelolipomas are benign tumors composed of an admixture of mature adipose tissue and normal hematopoietic cells. The vast majority occur within the adrenal glands, but several extra-adrenal myelolipomas (EAMLs) have been reported. The typical EAML is a solitary, well-defined mass within the abdomen, most commonly in the retroperitoneal presacral area. EAMLs may produce symptoms related to their mass effect, but they are occasionally incidental findings. Most commonly, the patient is older than 40 years and has no hematologic abnormalities. It is important to distinguish EAMLs from other soft tissue tumors, in particular liposarcomas, myxoid malignant fibrous histiocytomas, and extramedullary hematopoietic tumors. We discuss two cases of EAML. The first was in the retroperitoneum of a 76-year-old woman. It is the largest EAML ever reported, measuring 26 cm x 15 cm x 11 cm. The second, a presacral mass in a 68-year-old man, was diagnosed preoperatively by percutaneous computed tomography-guided fine needle aspiration biopsy. Preoperative diagnosis facilitated surgical management in this case. We discuss the clinical, radiologic, and pathologic characteristics and the differential diagnosis of this rare entity.


Analysis of HIV-1 replication block due to substitutions at F61 residue of reverse transcriptase reveals additional defects involving the RNase H function.
We reported previously that substitutions F61L, F61W, F61Y and F61A in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase affect strand displacement synthesis [T. S. Fisher, T. Darden and V. R. Prasad (2003) J. Mol. Biol., 325, 443-459]. We have now determined the effect of these mutations on HIV replication. All mutant viruses were replication defective. Measuring replication intermediates in infected cells did not reveal a specific block as all mutants displayed reduced DNA synthesis (wild-type>F61L>F61W>F61Y>F61A). Analysis of 2-LTR circle junctions revealed that F61W and F61Y mutants generated increased aberrant circle junctions. Circle junctions corresponding to F61Y included 3'-PPT insertions suggesting ribonuclease H defect. In vitro assays mimicking PPT primer generation indicated that F61L, F61W and F61Y mutant RTs were unaffected, while F61A mutant cleaved both at PPT/U3 junction and at +6 with similar efficiencies. In assays measuring cleavage at the RNA/DNA junction to remove the PPT primer, all mutants were significantly affected with F61Y and F61A being most severely impaired. Our results show that (i) replication block of most mutants is due to more than one biochemical defect; (ii) mutations in polymerase domain can affect the function of a distal domain; and (iii) virological analyses of RT mutations can yield insight into structure-function relationship that is otherwise not obvious.


An Optical Fiber-Based Data-Driven Method for Human Skin Temperature 3-D Mapping.
Human skin temperature mapping provides abundant information of physiological conditions of human body, which provides supplementary or alternative indicators for disease monitoring or diagnosis. The existing models of temperature mapping or temperature field distribution of human skin are generally established by finite element method. Due to the complexity of biological systems, it is challenging to achieve high accuracy mathematical models of temperature field of human skin. The goal of this study is to establish human skin temperature three-dimensional (3-D) mapping platform by integrating optical fibers and improved genetic algorithm-back propagation (GA-BP) neural network. The proposed data-driven method is capable of acquiring entire human skin temperature 3-D mapping by simply measuring a few points on human skin. Multiple experiments were conducted to validate the proposed method on different areas of human skin in different ambient environments. In each experiment setting, the measured data and the model output data were compared. The mean absolute error in all the validation experiments is 0.11 °C, which is lower than that in the state of the art using physical modeling for skin temperature prediction and more close to clinical accuracy. The results show that the proposed approach is accurate and reliable, which may provide a platform technology for human skin temperature mapping that can be used in both medical and scientific studies as well as home monitoring.


Transcriptome signature identifies distinct cervical pathways induced in lipopolysaccharide-mediated preterm birth.
With half a million babies born preterm each year in the USA and about 15 million worldwide, preterm birth (PTB) remains a global health issue. Preterm birth is a primary cause of infant morbidity and mortality and can impact lives long past infancy. The fact that there are numerous, and many currently unidentified, etiologies of PTB has hindered development of tools for risk evaluation and preventative therapies. Infection is estimated to be involved in nearly 40% of PTBs of known etiology; therefore, understanding how infection-mediated inflammation alters the cervical milieu and leads to preterm tissue biomechanical changes are questions of interest. Using RNA-seq, we identified enrichment of components involved in inflammasome activation and unique proteases in the mouse cervix during lipopolysaccharide (LPS)-mediated PTB and not physiologically at term before labor. Despite transcriptional induction of inflammasome components, there was no evidence of functional activation based on assessment of mature IL1B and IL18 proteins. The increased transcription of proteases that target both elastic fibers and collagen and concentration of myeloid-derived cells capable of protease synthesis in the cervical stroma support the structural disruption of elastic fibers as a functional output of protease activity. The recent demonstration that elastic fibers contribute to the biomechanical function of the pregnant cervix suggests their protease-induced disruption in the infection model of LPS-mediated PTB and may contribute to premature loss of mechanical competency and preterm delivery. Collectively, the transcriptomics and ultrastructural data provide new insights into the distinct mechanisms of premature cervical remodeling in response to infection.


Structural basis for the interference between nicorandil and sulfonylurea action.
Nicorandil is a new antianginal agent that potentially may be used to treat the cardiovascular side effects of diabetes. It is both a nitric oxide donor and an opener of ATP-sensitive K(+) (K(ATP)) channels in muscle and thereby causes vasodilation of the coronary vasculature. The aim of this study was to investigate the domains of the K(ATP) channel involved in nicorandil activity and to determine whether nicorandil interacts with hypoglycemic sulfonylureas that target K(ATP) channels in pancreatic beta-cells. K(ATP) channels in muscle and beta-cells share a common pore-forming subunit, Kir6.2, but possess alternative sulfonylurea receptors (SURs; SUR1 in beta-cells, SUR2A in cardiac muscle, and SUR2B in smooth muscle). We expressed recombinant K(ATP) channels in Xenopus oocytes and measured the effects of drugs and nucleotides by recording macroscopic currents in excised membrane patches. Nicorandil activated Kir6.2/SUR2A and Kir6.2/SUR2B but not Kir6.2/SUR1 currents, consistent with its specificity for cardiac and smooth muscle K(ATP) channels. Drug activity depended on the presence of intracellular nucleotides and was impaired when the Walker A lysine residues were mutated in either nucleotide-binding domain of SUR2. Chimeric studies showed that the COOH-terminal group of transmembrane helices (TMs), especially TM 17, is responsible for the specificity of nicorandil for channels containing SUR2. The splice variation between SUR2A and SUR2B altered the off-rate of the nicorandil response. Finally, we showed that nicorandil activity was unaffected by gliclazide, which specifically blocks SUR1-type K(ATP) channels, but was severely impaired by glibenclamide and glimepiride, which target both SUR1 and SUR2-type K(ATP) channels.


Antifungal activity of Lavandula angustifolia essential oil against Candida albicans yeast and mycelial form.
The antifungal activity of the essential oil of Lavandula angustifolia Mill. (lavender oil) and its main components, linalool and linalyl acetate, was investigated against 50 clinical isolates of Candida albicans (28 oropharyngeal strains, 22 vaginal strains) and C. albicans ATCC 3153. Growth inhibition, killing time and inhibition of germ tube formation were evaluated. The chemical composition of the essential oil was determined by gas chromatography and mass spectrometry. Lavender oil inhibited C. albicans growth: mean minimum inhibitory concentration (MIC) of 0.69% (vol./vol.) (vaginal strains) and 1.04% (oropharyngeal strains); mean MFC of 1.1% (vaginal strains) and 1.8% (oropharyngeal strains). Linalool was more effective than essential oil: mean MIC of 0.09% (vaginal strains) and 0.29% (oropharyngeal strains); mean MFC of 0.1% (vaginal strains) and 0.3% (oropharyngeal strains). Linalyl acetate was almost ineffective. Lavender oil (2%) killed 100% of the C. albicans ATCC 3153 cells within 15 min; linalool (0.5%) killed 100% of the cells within 30 s. The essential oil inhibited germ tube formation (mean MIC of 0.09%), as did the main components (MIC of 0.11% for linalool and 0.08% for linalyl acetate). Both the essential oil and its main components inhibited hyphal elongation of C. albicans ATCC 3153 (about 50% inhibition at 0.016% with each substance). Lavender oil shows both fungistatic and fungicidal activity against C. albicans strains. At lower concentrations, it inhibits germ tube formation and hyphal elongation, indicating that it is effective against C. albicans dimorphism and may thus reduce fungal progression and the spread of infection in host tissues.


Osteoradionecrosis of the mandible: a review.
Osteoradionecrosis (ORN) is a complex metabolic and tissue homeostatic deficiency created by radiation-induced tissue injury. Although infection was initially felt to play an important role in its pathogenesis, the role of microorganisms has been relegated to that of a contaminant. Trauma in some cases may provide a possible initiating factor for the process, or in the case of spontaneous ORN, the use of radioactive implants and the higher doses of radiation therapy as practised in the megavoltage era, may have an influence. The role of prevention cannot be over-emphasized. Dental evaluation is an important part of the initial patient assessment. Preservation of good or average teeth is recommended with subsequent regular follow-up and dental care. Post-treatment dental extraction should be minimized and delayed if possible until at least nine-12 months after completion of radiation therapy. Management of ORN requires assessment of the extent, both clinically and radiologically. Conservative management is recommended unless the disease is advanced or progressive. Radiologic evidence of fracture requires surgery. Modern surgical techniques offer satisfactory reconstruction. The role of hyperbaric oxygen remains controversial and limited to centers where hyperbaric techniques exist. The treatment is complex and time-consuming and results are confounded by the concurrent use of local antiseptic/antibiotic measures and surgery.


No evidence for temporal decay in working memory.
What drives forgetting in working memory? Recent evidence suggests that in a complex-span task in which an irrelevant processing task alternates with presentation of the memoranda, recall declines when the time taken to complete the processing task is extended while holding the time for rehearsal in between processing steps constant (Portrat, Barrouillet, & Camos, 2008). This time-based forgetting was interpreted in support for the role of time-based decay in working memory. In this article, we argue the contrary position by (a) showing in an experiment that the processing task in Portrat et al.'s (2008) study gave rise to uncontrolled post-error processes that occupied the attentional bottleneck, thus preventing restorative rehearsal, and (b) showing that when those post-error processes are statistically controlled, there is no evidence for temporal decay in Portrat et al.'s study. We conclude that currently there exists no direct evidence for temporal decay in the complex-span paradigm.


Oto-endoscopy: a reliable and validated technique for phenotyping otitis media in the mouse.
The mouse is a widely used model for investigating the pathophysiological and genetic bases of otitis media (OM). It has proven a valuable tool for investigating the multifactorial bases of OM including the role of pathogens, anatomical factors, inflammatory mediators and susceptibility loci. However, straightforward and robust phenotyping tools for identifying murine otitis media are lacking, which has precluded for example the identification of mice with OM in genetic screens without resorting to time-consuming histopathology. We have set out to develop a phenotyping platform for the detection of OM in mice utilizing oto-endoscopy. We have applied the technique to a cohort of mice genetically susceptible to chronic otitis media. We show that oto-endoscopy is a safe, reliable and valid method for detecting otitis media in the mouse and discuss its utility in screens to identify novel genes involved with susceptibility to OM.


Genome Mining-Mediated Discovery of a New Avermipeptin Analogue in Streptomyces actuosus ATCC 25421.
Streptomyces actuosus ATCC 25421 was famous for producing thiopeptide nosiheptide, which has widely been used as a feed additive for the promotion of animal growth. Herein, we report the complete genome sequence of S. actuosus ATCC 25421, which consists of an 8,145,579-bp circular chromosome with a G+C content of 72.53 % containing 7 536 protein-coding genes. The antiSMASH 3.0 program was used to identify 49 biosynthetic gene clusters for putative secondary metabolites, including a putative lantipeptide gene cluster that showed 85 % similarity to the reported informatipeptin biosynthetic gene cluster, indicating that the putative lantipeptide gene cluster has the ability to generate the informatipeptin analogue. Compared with avermipeptin, the lantipeptide precursor peptide (termed avermipeptin B) from S. actuosus ATCC 25421 contains a 14-aa leader peptide and a 24-aa core peptide, in which Ile15 was different from Val15 in avermipeptin. We also deduced the structure and the biosynthetic mechanism of avermipeptin B. Heterologous expression of the avermipeptin B biosynthetic gene cluster in S. lividans TK24 was characterized by high-resolution mass spectrometry (ESI-MS/MS). Finally, we found that avermipeptin B displayed strong activity against Gram-positive strains. The genome sequence reported here can encourage us to mine novel secondary metabolites and investigate their biosynthetic mechanism in the future.


Genome-wide association meta-analysis highlights light-induced signaling as a driver for refractive error.
Refractive errors, including myopia, are the most frequent eye disorders worldwide and an increasingly common cause of blindness. This genome-wide association meta-analysis in 160,420 participants and replication in 95,505 participants increased the number of established independent signals from 37 to 161 and showed high genetic correlation between Europeans and Asians (>0.78). Expression experiments and comprehensive in silico analyses identified retinal cell physiology and light processing as prominent mechanisms, and also identified functional contributions to refractive-error development in all cell types of the neurosensory retina, retinal pigment epithelium, vascular endothelium and extracellular matrix. Newly identified genes implicate novel mechanisms such as rod-and-cone bipolar synaptic neurotransmission, anterior-segment morphology and angiogenesis. Thirty-one loci resided in or near regions transcribing small RNAs, thus suggesting a role for post-transcriptional regulation. Our results support the notion that refractive errors are caused by a light-dependent retina-to-sclera signaling cascade and delineate potential pathobiological molecular drivers.


Impact of Genetic Variability on Physiological Responses to Caffeine in Humans: A Systematic Review.
Emerging research has demonstrated that genetic variation may impact physiological responses to caffeine consumption. The purpose of the present review was to systematically recognize how select single nucleotide polymorphisms (SNPs) impact habitual use of caffeine as well as the ergogenic and anxiogenic consequences of caffeine. Two databases (PubMed and EBSCO) were independently searched using the same algorithm. Selected studies involved human participants and met at least one of the following inclusion criteria: (a) genetic analysis of individuals who habitually consume caffeine; (b) genetic analysis of individuals who underwent measurements of physical performance with the consumption of caffeine; (c) genetic analysis of individuals who underwent measurements of mood with the consumption of caffeine. We included 26 studies (10 randomized controlled trials, five controlled trials, seven cross-sectional studies, three single-group interventional studies and one case-control study). Single nucleotide polymorphisms in or near the cytochrome P450 (CYP1A2) and aryl hydrocarbon receptor (AHR) genes were consistently associated with caffeine consumption. Several studies demonstrated that the anxiogenic consequences of caffeine differed across adenosine 2a receptor (ADORA2A) genotypes, and the studies that investigated the effects of genetic variation on the ergogenic benefit of caffeine reported equivocal findings (CYP1A2) or warrant replication (ADORA2A).


Pyrvinium Targets CD133 in Human Glioblastoma Brain Tumor-Initiating Cells.
Clonal evolution of cancer may be regulated by determinants of stemness, specifically self-renewal, and current therapies have not considered how genetic perturbations or properties of stemness affect such functional processes. Glioblastoma-initiating cells (GICs), identified by expression of the cell surface marker CD133, are shown to be chemoradioresistant. In the current study, we sought to elucidate the functional role of CD133 in self-renewal and identify compounds that can specifically target this CD133(+) treatment-refractory population. Using gain/loss-of-function studies for CD133 we assessed the in vitro self-renewal and in vivo tumor formation capabilities of patient-derived glioblastoma cells. We generated a CD133 signature combined with an in silico screen to find compounds that target GICs. Self-renewal and proliferation assays on CD133-sorted samples were performed to identify the preferential action of hit compounds. In vivo efficacy of the lead compound pyrvinium was assessed in intracranial GIC xenografts and survival studies. Lastly, microarray analysis was performed on pyrvinium-treated GICs to discover core signaling events involved. We discovered pyrvinium, a small-molecule inhibitor of GIC self-renewal in vitro and in vivo, in part through inhibition of Wnt/β-catenin signaling and other essential stem cell regulatory pathways. We provide a therapeutically tractable strategy to target self-renewing, chemoradioresistant, and functionally important CD133(+) stem cells that drive glioblastoma relapse and mortality. Our study provides an integrated approach for the eradication of clonal populations responsible for cancer progression, and may apply to other aggressive and heterogeneous cancers.


Assessment of DNA damage in ceramic workers.
It is known that ceramic workers are potentially exposed to complex mixture of chemicals such as silica, inorganic lead, lime, beryllium and aluminum that can be associated with an increased risk of several diseases. All operations in the ceramic industries such as mixing, moulding, casting, shaking out and finishing jobs, have been associated with the higher exposure levels and in most of the silica-related industries, average overall exposure exceeded permissible exposure levels for respirable crystalline silica. The aim of this study was to evaluate the possible genotoxic damage in ceramic workers exposed to complex mixture of chemicals mainly crystalline silica. For this purpose, the blood and buccal epithelial cell samples were taken from the ceramic workers (n = 99) and their controls (n = 81). The genotoxicity was assessed by the alkaline comet assay in isolated lymphocytes and whole blood. Micronucleus (MN), binucleated (BN), pyknotic (PYC), condensed chromatin (CC), karyolytic (KYL), karyorrhectic (KHC) and nuclear bud (NBUD) frequencies in buccal epithelial cells and plasma 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) levels were also evaluated. In the study, 38 workers were diagnosed with silicosis, 9 workers were suspected to have silicosis, whereas 52 workers were found to be healthy. DNA damage in blood and lymphocytes; MN, CC + KHC, PYC frequencies in buccal epithelial cells and 8-oxodG levels in plasma were increased in workers compared to their controls. These results showed that occupational chemical mixture exposure in ceramic industry may cause genotoxic damage that can lead to important health problems in the workers.


Aesthetic management of the nasal component of naso-orbital ethmoid fractures.
After studying this article, the participant should be able to: 1. Discuss the critical anatomic structures of the nose that are affected in naso-orbital ethmoid fractures. 2. Discuss the advantages of early, complete nasal reconstruction of these fractures. 3. Apply a clinical algorithm to such nasal reconstruction. 4. List the techniques used in aesthetic reconstruction of the nose in naso-orbital ethmoid fractures. Fractures of the naso-orbital ethmoid complex pose challenging management issues. Although basic treatment principles have been well described, the aesthetic management of the nasal component has not been adequately addressed in the literature. When secondary nasal deformities occur, they are difficult to correct. Optimal primary correction of the nasal deformity is accomplished using the following four principles: (1) rigid fixation of the nasal pyramid and restoration of nasal height and length; (2) restoration of tip projection; (3) septal reduction/reconstruction; and (4) lateral nasal wall augmentation. Successful management of naso-orbital ethmoid fractures is a complex and challenging task. Both the bony and soft-tissue components must be addressed and the extent of the injury must be adequately diagnosed to avoid omission of critical steps in the reconstruction. Inadequate treatment of naso-orbital ethmoid fractures can produce a severe cosmetic deformity that is very difficult to correct secondarily. The authors discuss the nasal component of naso-orbital ethmoid complex injuries and detail the key principles in their algorithm for aesthetic nasal reconstruction.


Transcriptional patterns of the pCD41 (U27) locus of human herpesvirus 6.
Human herpesvirus 6 (HHV-6) is a lymphotropic herpesvirus, and in vitro, it can productively infect many of the same cell types that human immunodeficiency virus (HIV) infects. Simultaneous infection of T cells by HIV and HHV-6 can lead to both activation of the HIV promoter and acceleration of the cytopathic effects. Several HHV-6 genes have been demonstrated to activate HIV promoter expression. Among them is a cDNA clone, pCD41 (U27), which codes for the HHV-6 DNA polymerase accessory protein. We have now further characterized the transcription pattern in the pCD41 locus and identified at least six RNA species, ranging in size from 1.2 to 4.5 kb. Northern (RNA) blot analyses showed no significant difference in RNA patterns between the HHV-6 variant A (GS) and variant B (Z29) viruses. All the RNA species detected by pCD41 are polyadenylated and polyribosome associated, suggesting that they may be actively engaged in protein synthesis. Cycloheximide and phosphonoacetic acid inhibition assay results indicate that all the pCD41 RNA species belong to the herpesviral early-late family. Using primer extension and S1 mapping techniques, the 5' and 3' ends of each transcript were mapped to different positions, and no splicing was observed.


Establishing lignin structure-upgradeability relationships using quantitative 1H-13C heteronuclear single quantum coherence nuclear magnetic resonance (HSQC-NMR) spectroscopy.
Lignin depolymerization could provide an attractive renewable aromatic feedstock for the chemical industry. Past studies have suggested that lignin structural features such as ether content are correlated to lignin's upgradeability. An obstacle to the development of a conclusive causal relationship between lignin structure and upgradeability has been the difficulty to quantitatively measure lignin structural features. Here, we demonstrated that a modified HSQC-NMR method known as HSQC0 can accurately quantify lignin functionalities in extracted lignin using several synthetic polymer models. We then prepared a range of isolated lignin samples with a wide range of ether contents (6-46%). By using a simple ether cleavage model, we were able to predict final depolymerization yields very accurately (<4% error), conclusively demonstrating the direct causal relationship between ether content and lignin activity. The accuracy of this model suggests that, unlike in native lignin, ether linkages no longer appear to be randomly distributed in isolated lignin.


Parental understanding of intoeing gait - A preliminary study.
Evaluation of a child's intoeing gait is one of the most common referrals made to a pediatric orthopedist. Families may have difficulty understanding the often transient and usually benign nature of intoeing. The purpose of this study was to investigate parental perceptions of an intoeing gait both before and after consultation with an orthopedic practitioner. 48 parents of children referred to pediatric orthopedic surgeons for evaluation of intoeing gait completed a 22-item questionnaire that assessed demographics, anxiety, and parental perceptions of intoeing. Questionnaires were administered before and after visits. Statistical analysis was performed using Wilcoxon signed-rank tests and Fisher exact tests. Before their visits, parents reported similar levels of anxiety and understanding regarding intoeing, independent of their age or education. After the visits, anxiety decreased (P < 0.001), and understanding increased (P < 0.001) although younger parents (18-25 years) reported higher post-visit anxiety compared to parents older than 25 (P = 0.014). Similarly, parents with a high school degree or less reported higher post-visit anxiety compared to parents who attended college (P = 0.009). Post-visit understanding was inversely correlated with post-visit anxiety (r = -0.717; P < 0.001). Additionally, parents who reported high anxiety post-visit stated they were more likely to seek additional care (P < 0.001). Younger parents with lower education levels were more likely to leave visits with high anxiety and poor understanding. These parents were more likely to consider seeking further treatment for their child's intoeing, placing additional stress on their child, themselves and an overburdened healthcare system.


A highly efficient Friedel-Crafts reaction of 3-hydroxyoxindoles and aromatic compounds to 3,3-diaryl and 3-alkyl-3-aryloxindoles catalyzed by Hg(ClO4)2·3H2O.
We report a highly efficient Friedel-Crafts reaction of 3-alkyl or 3-aryl 3-hydroxyoxindoles with a variety of aromatic and heteroaromatic compounds to unsymmetrical 3,3-diaryloxindoles or 3-alkyl-3-aryloxindoles, which are interesting medicinal targets and useful building blocks for the synthesis of natural products. Hg(ClO(4))(2)·3H(2)O was identified as a powerful catalyst for this reaction, and is significantly more efficient than other screened metal perchlorate hydrates and Brønsted acids such as HOTf and HClO(4). The high catalytic property of Hg(ClO(4))(2)·3H(2)O originates from the unprecedented dual activation effects of aromatic mercuration, which could generate a strong protic acid to facilitate the generation of a carbocation at the C3-position of oxindoles and simultaneously form the more reactive nucleophilic reaction partner.


Electrical superior vena cava isolation using photodynamic therapy in a canine model.
With the new era of multi-tip radiofrequency or balloon ablation catheters replacing the point-to-point ablation strategy, we aimed to determine the feasibility of a ring-laser catheter ablation technology to electrically isolate the superior vena cava (SVC) by exploring the advantages of the limitless catheter tip size possibly with the photodynamic therapy (PDT)-mediated ablation. We developed a first-generation prototype of a circular-laser-mapping catheter by fitting a 7 cm plastic optical fibre onto a circular variable-loop Lasso™ mapping catheter. Following SVC venography, both the laser catheter and another ring catheter for monitoring the SVC potentials were placed at the SVC. After the systemic infusion of a photosensitizer (talaporfin sodium), we initiated the irradiation with an output of 1 W in three canines and 0.3 W in four. The creation of electrical isolation as well as occurrence of phrenic nerve injury, sinus node injury, and SVC stenosis were evaluated before, immediately after, and 1 month after the procedure. A PDT-mediated SVC isolation was successfully performed in all seven canines. The isolation was completed with a laser irradiation of 70.4 ± 71.4 J/cm under 30.9 ± 5.0 µg/mL of a photosensitizer without any sinus node injury, phrenic nerve palsy, or SVC stenosis in both the acute and chronic evaluations. The minimum isolation time of 270 s was not correlated with the laser input power or the photosensitizer concentration. The electrical SVC isolation was successfully and instantly achieved using the PDT laser-ring catheter without any complications.


Modulation of Aphid Vector Activity by Potato virus Y on In Vitro Potato Plants.
The effects of the infection of potato (Solanum tuberosum) plants by the nonpersistent Potato virus Y (PVY) were studied on the host plant colonization behavior of different colonizing (Myzus persicae) and noncolonizing (Aphis fabae, Brevicoryne brassicae, and Sitobion avenae) aphid species. The underlying questions of this study were to know how aphids respond when faced with PVY-infected plants and whether plant infection can modify the aphid behavior involved in PVY spread. Short-range orientation behavior was observed using a dual-choice set-up and aphid feeding behavior was monitored using the electrical penetration graph technique. None of the aphid species discriminated between healthy and PVY-infected plants. Nevertheless, most individuals of M. persicae landed on and probed only in one plant whereas noncolonizing aphid species exhibited interplant movements. Study of the aphid feeding behavior showed that PVY infection essentially modified phloem and xylem ingestion. M. persicae and S. avenae exhibited an increased duration of phloem phases on PVY-infected plants whereas A. fabae showed a decreased duration of phloem phases that benefited from an increased duration of xylem ingestion phases. None of these parameters were changed in B. brassicae. These data present evidence that aphids can respond to plants infected by nonpersistent viruses. Such behavioral modifications are discussed within the context of PVY spread in potato crops.


Hopeful struggling for health: Experiences of participating in computerized cognitive training and aerobic training for persons with stress-related exhaustion disorder.
It is important to understand how people with exhaustion disorder (ED) perceive interventions aiming to facilitate cognitive functioning. Therefore, the overall aim of this study was to explore experiences from persons with ED after participating in a 12-week intervention of either computerized cognitive training or aerobic training. Both interventions were performed in addition to a multimodal rehabilitation programme. Thirteen participants, 11 women and 2 men, were interviewed about pros and cons with participating in the training. The interviews were analysed with Qualitative Content Analysis. The analyses resulted in the theme hopeful struggling for health and the categories support, motivation and sensations. It was hard work recovering from ED. Support from others who are in the same situation, family members, and technology and routines for the training were strongly emphasized as beneficial for recovery. Timing, i.e., matching activities to the rehabilitation programme, getting feedback and perceiving joy in the training were important for motivation. Participants in both interventions experienced positive sensations with improved memory performance, everyday life functioning and increased faith in the prospect of recovery. However, it is important to consider various aspects of support and motivation in both computerized cognitive training and aerobic training to enable participants to pursue their participation.


Surgical treatment of 127 anterior choroidal artery aneurysms: a cohort study of resultant ischemic complications.
The anterior choroidal artery (AChA) supplies important areas of the nervous system, particularly the posterior limb of the internal capsule and optic radiation. Treatment of AChA aneurysms poses particular challenges because of the complex anatomy of the aneurysm associated with the relatively small diameter of AChAs, making preservation of the parent vessel during clip ligation or endosaccular coiling challenging. To investigate the incidence and features of ischemia in treatment of AChA aneurysms. A prospectively maintained database of patients who underwent treatment of aneurysms from 1985 to 2011 was queried to find patients with AChA aneurysms. Age, sex, Hunt and Hess grade, treatment modality, and complications were analyzed by use of the unpaired Student t test and Fisher exact test. One hundred twenty-two patients harbored 127 AChA aneurysms, and 67% (82 of 122) had multiple aneurysms. Treatment included 112 microsurgical clip ligations, 8 endosaccular coil embolizations, 5 aneurysmal wrappings, and 2 surgical explorations. Complications developed in 53% (67 of 127) of AChA aneurysms. Postoperative ischemia occurred in 12% (15 of 127) of treated aneurysms. The number of temporary clip applications was most closely associated with postoperative ischemia. Glasgow Outcome Scale scores of 4 or 5 were obtained by 78% at discharge, 89% at 6 months, and 85% at 1 year. The ischemic complication rate from surgical treatment of AChA aneurysms is most closely associated with higher frequency of temporary clip applications for proximal control and may be lower than previously reported. Supplementary intraoperative tools and limitation of vessel manipulation should be used to improve outcomes.


Microscope-based label-free microfluidic cytometry.
A microscope-based label-free microfluidic cytometer capable of acquiring two dimensional light scatter patterns from single cells, pattern analysis of which determines cellular information such as cell size, orientation and inner nanostructure, was developed. Finite-difference time-domain numerical simulations compared favorably with experimental scatter patterns from micrometer-sized beads and cells. The device was capable of obtaining light scattering patterns from the smallest mature blood cells (platelets) and cord blood hematopoietic stem/progenitor cells (CD34 + cells) and myeloid precursor cells. The potential for evaluation of cells using this label-free microfluidic cytometric technique was discussed.


Improving surveillance for pediatric Clostridium difficile infection: derivation and validation of an accurate case-finding tool.
The incidence of Clostridium difficile infection (CDI) is increasing. Multicenter studies of CDI have been limited by the lack of valid case-finding tools. To facilitate pediatric studies of CDI, we constructed a case-finding tool using administrative data. A cross-sectional study was performed using the Pediatric Health Information System database and microbiologic data from 4 member hospitals. Using patients with laboratory-confirmed CDI as the standard, we determined the sensitivity, specificity, positive (PPV), and negative (NPV) predictive value of an ICD-9-CM code for identifying children with laboratory-confirmed CDI. We identified 109 patients with laboratory-confirmed CDI and 119 patients with CDI ICD-9-CM code. The sensitivity, specificity, PPV, and NPV were 80.73%, 99.89%, 73.95%, and 99.92%, respectively, for this comparison. The addition of a billing charge for both C. difficile laboratory test and treatment medication to the ICD-9-CM code increased the specificity and PPV, but resulted in a slight decrease in the sensitivity and NPV. The use of administrative data for identifying pediatric cases of CDI was also compared with that of chart review, and was found to be a stronger surrogate for identifying cases of CDI when compared with microbiology data alone. These results demonstrate that the use of administrative data for CDI is a reliable and accurate method for identifying pediatric patients with CDI. The use of administrative data could facilitate the completion of larger studies due to its greater accessibility and reduced costs.


Not just about sunburn--the ozone hole's profound effect on climate has significant implications for Southern Hemisphere ecosystems.
Climate scientists have concluded that stratospheric ozone depletion has been a major driver of Southern Hemisphere climate processes since about 1980. The implications of these observed and modelled changes in climate are likely to be far more pervasive for both terrestrial and marine ecosystems than the increase in ultraviolet-B radiation due to ozone depletion; however, they have been largely overlooked in the biological literature. Here, we synthesize the current understanding of how ozone depletion has impacted Southern Hemisphere climate and highlight the relatively few documented impacts on terrestrial and marine ecosystems. Reviewing the climate literature, we present examples of how ozone depletion changes atmospheric and oceanic circulation, with an emphasis on how these alterations in the physical climate system affect Southern Hemisphere weather, especially over the summer season (December-February). These potentially include increased incidence of extreme events, resulting in costly floods, drought, wildfires and serious environmental damage. The ecosystem impacts documented so far include changes to growth rates of South American and New Zealand trees, decreased growth of Antarctic mosses and changing biodiversity in Antarctic lakes. The objective of this synthesis was to stimulate the ecological community to look beyond ultraviolet-B radiation when considering the impacts of ozone depletion. Such widespread changes in Southern Hemisphere climate are likely to have had as much or more impact on natural ecosystems and food production over the past few decades, than the increased ultraviolet radiation due to ozone depletion.


Transient structural variations have strong effects on quantitative traits and reproductive isolation in fission yeast.
Large structural variations (SVs) within genomes are more challenging to identify than smaller genetic variants but may substantially contribute to phenotypic diversity and evolution. We analyse the effects of SVs on gene expression, quantitative traits and intrinsic reproductive isolation in the yeast Schizosaccharomyces pombe. We establish a high-quality curated catalogue of SVs in the genomes of a worldwide library of S. pombe strains, including duplications, deletions, inversions and translocations. We show that copy number variants (CNVs) show a variety of genetic signals consistent with rapid turnover. These transient CNVs produce stoichiometric effects on gene expression both within and outside the duplicated regions. CNVs make substantial contributions to quantitative traits, most notably intracellular amino acid concentrations, growth under stress and sugar utilization in winemaking, whereas rearrangements are strongly associated with reproductive isolation. Collectively, these findings have broad implications for evolution and for our understanding of quantitative traits including complex human diseases.


Propagation of spoof surface plasmon on metallic square lattice: bending and splitting of self-collimated beams.
The propagation characteristics of spoof surface plasmon modes are studied in both real and reciprocal spaces. From the metallic square lattice, we obtain constant frequency contours by directly measuring electric fields in the microwave frequency regime. The anisotropy of the measured constant frequency contour supports the presence of the negative refraction and the self-collimation which are confirmed from measured electric fields. Additionally, we demonstrate the spoof surface plasmon beam splitter in which the splitting ratio of the self-collimated beam is controlled by varying the height of rods.


The nicotinic acid receptor--a new mechanism for an old drug.
Non-esterified fatty acids in plasma originate from adipose tissue. Delivery of fatty acids to the liver provides the substrate for VLDL triglycerides. Insulin-sensitive organs, overburdened by high concentrations of non-esterified fatty acids, may develop resistance to insulin action. In addition, insulin secretion from pancreatic beta-cells may be impaired by long-standing elevation of concentrations of non-esterified fatty acid in plasma. Normally, such concentrations fluctuate over the day depending on the transient suppression of lipolysis from adipose tissue by insulin released after meals. Diurnal concentrations of non-esterified fatty acid are often elevated in obesity, in particular in male-pattern upper-body fat accumulation. Nicotinic acid is the only drug that primarily lowers concentrations of non-esterified fatty acids and thereby lowers VLDL triglycerides. Nicotinic acid, or its analogues, seems to alleviate insulin resistance in the short-term whereas, paradoxically, the long-term effect is often the opposite. Suppression of lipolysis by nicotinic acid gives rise to a prominent rebound and the degree to which this occurs might explain this paradox. The exact cellular mechanism by which nicotinic acid exerts its antilipolytic effects has not been known until the recent discovery of a distinct G-protein coupled receptor. Nicotinic acid is a high affinity ligand, but the endogenous ligand is still unknown. Recently, Tina Rubic and colleagues (Biochem Pharmacol 2004; 67: 411-19) proposed a mechanism in which nicotinic acid stimulates cholesterol mobilisation from macrophages, thereby providing a potential link between regression of atherosclerosis and use of nicotinic acid. Research on signalling through the nicotinic acid receptor might give rise to novel and more effective methods to interfere with fatty-acid metabolism, with insulin resistance, hyperlipidaemia, and atherosclerosis as target diseases.


[A prospective, randomized, open and comparative study to evaluate the safety and efficacy of blue light treatment versus a topical benzoyl peroxide 5% formulation in patients with acne grade II and III].
Many acne patients improve after exposure to sunlight and there are many reports about the efficacy of blue light phototherapy on acne lesions. The purpose of this study was to evaluate efficacy and safety of blue light treatment versus topical benzoyl peroxide 5% formulation in patients with acne grades II and III. Sixty volunteers with facial acne were included and evaluated in 5 visits: the first one for screening, another 3 held on days 7, 14 and 28 of treatment, and the last one after 14 days of the end of treatment. Thirty of them were irradiated with Blue Light (8 times, twice a week) and the other thirty were treated with topical Benzoyl Peroxide 5% formulation, auto-applied twice a day, every day. We assessed the severity of acne by counting the lesions and analyzing the photographs. The improvement achieved by the blue light was the same as the one with benzoyl peroxide, regardless of the type of lesion (p 0.05). Otherwise, the side effects were less frequent in the group treated with blue light. Blue light irradiation was as effective as benzoyl peroxide in acne treatment grades II and III but there were fewer side effects.


Estrogen and cigarette sidestream smoke particulate matter exhibit ERα-dependent tumor-promoting effects in lung adenocarcinoma cells.
Estrogen and secondhand smoke are key risk factors for nonsmoking female lung cancer patients who frequently have lung adenocarcinoma and show tumor estrogen receptor α (ERα) expression. We speculated that estrogen and secondhand smoke might cause harmful effects via ERα signaling. Our results showed that 17β-estradiol (E2), the primary form of endogenous estrogen, exacerbated proliferation, migration, and granzyme B resistance of lung adenocarcinoma cells in an ERα-dependent manner. Cigarette sidestream smoke particulate matter (CSSP), the major component of secondhand smoke, could activate ERα activity dose dependently in human lung adenocarcinoma cells. The estrogenic activity of CSSP was abolished by an ERα-selective antagonist. CSSP regulated the nuclear entry, phosphorylation, and turnover of ERα similarly to E2. Furthermore, CSSP enhanced E2-stimulated ERα activity and Ser118 phosphorylation even when ERα became saturated with E2. Activation of ERα by CSSP required GSK3β activity, but not involving polycyclic aromatic hydrocarbons, reactive oxygen species, calcium, epidermal growth factor receptor, and PI3K/Akt. Although CSSP possessed cytotoxicity, ERα-expressing cells grew and migrated faster than nonexpressing cells on recovery from CSSP exposure as observed in E2-pretreated cells. Knockdown of ERα by siRNA diminished E2- and CSSP-stimulated cell migration. Twenty-one genes, including SERPINB9, were identified to be upregulated by both E2 and CSSP via ERα. Increased SERPINB9 expression was accompanied with increased resistance to granzyme B-mediated apoptosis. This study demonstrates that estrogen has ERα-dependent tumor-promoting activity. CSSP acts like estrogen and shows a potential to enhance estrogen-induced ERα action.


Development of a high-throughput arrayed neural circuitry platform using human induced neurons for drug screening applications.
Proper brain function relies on the precise arrangement and flow of information between diverse neural subtypes. Developing improved human cell-based models which faithfully mimic biologically relevant connectivity patterns may improve drug screening efforts given the limited success of animal models to predict safety and efficacy of therapeutics in human clinical trials. To address this need, we have developed experimental models of defined neural circuitries through the compartmentalization of neuronal cell subtypes in a 96 well plate-based platform where each microwell is divided into two compartments connected by microchannels allowing high-throughput screening (HTS) of small molecules. We demonstrate that we can generate subtype-specific excitatory and inhibitory induced neuronal cells (iNs) from human stem cell lines and that these neurons form robust functional circuits with defined connectivity. Through the use of the genetically encoded calcium indicator GCaMP6f, we monitor calcium ion transients generated during neuronal firing between and within compartments. We further demonstrate functionality of the circuit by perturbing network activity through the addition of glutamate receptor blockers using automated liquid handling. Lastly, we show that we can stimulate network activity in defined neuronal subtypes through the expression of the designer receptor exclusively activated by designer drugs (DREADD) hM3Dq and application of the ligand clozapine-N-oxide (CNO). Our results demonstrate the formation of functional neural circuits in a high-throughput platform that is compatible with compound screening, representing an important step towards developing new screening platforms for studying and ultimately treating psychiatric brain disorders that arise from disordered neural circuit function.


Postsynaptic excitability is necessary for strengthening of cortical sensory responses during experience-dependent development.
Sensory experience is necessary for normal cortical development. This has been shown by sensory deprivation and pharmacological perturbation of the cortex. Because these manipulations affect the cortical network as a whole, the role of postsynaptic cellular properties during experience-dependent development is unclear. Here we addressed the developmental role of somatodendritic excitability, which enables postsynaptic spike timing-dependent forms of plasticity, in rat somatosensory cortex. We used short interfering RNA (siRNA)-based knockdown of Na+ channels to suppress the somatodendritic excitability of small numbers of layer 2/3 pyramidal neurons in the barrel cortex, without altering the ascending sensory pathway. In vivo recordings from siRNA-expressing cells revealed that this manipulation interfered with the normal developmental strengthening of sensory responses. The sensory responsiveness of neighboring cortical neurons was unchanged, indicating that the cortical network was unchanged. We conclude that somatodendritic excitability of the postsynaptic neuron is needed for the regulation of synaptic strength in the developing sensory cortex.


Cardiac Disease after Pregnancy: A Growing Problem.
Pregnancy complications, such as hypertensive disorders or preterm delivery, identify families predisposed to cardiovascular problems at other times in life. Whether the pregnancy complication induces cardiac disease or whether the pregnancy stress unmasks an underlying predisposition remains unclear. However, improved survival following severe pregnancy complications for both the mother and, in particular, the offspring - who is often born preterm - has resulted in a growing cohort of individuals who carry this increased cardiovascular risk. Research to understand the underlying pathological mechanisms that link these conditions might ultimately lead to novel therapeutic or prevention strategies for both cardiovascular and pregnancy disease.


Mitochondrial DNA copy number and incident atrial fibrillation.
Mechanistic studies suggest that mitochondria DNA (mtDNA) dysfunction may be associated with increased risk of atrial fibrillation (AF). The association between mtDNA copy number (mtDNA-CN) and incident AF in the general population, however, remains unknown. We conducted prospective analyses of 19,709 participants from the Atherosclerosis Risk in Communities Study (ARIC), the Multi-Ethnic Study of Atherosclerosis (MESA), and the Cardiovascular Health Study (CHS). mtDNA-CN from the peripheral blood was calculated from probe intensities on the Affymetrix Genome-Wide Human single nucleotide polymorphisms (SNP) Array 6.0 in ARIC and MESA and from multiplexed real-time quantitative polymerase chain reaction (qPCR) in CHS. Incident AF cases were identified through electrocardiograms, review of hospital discharge codes, Medicare claims, and death certificates. The median follow-up time was 21.4 years in ARIC, 12.9 years in MESA, and 11.0 years in CHS, during which 4021 participants developed incident atrial fibrillation (1761 in ARIC, 790 in MESA, and 1470 in CHS). In fully adjusted models, participants with the lowest quintile of mitochondria DNA copy number had an overall 13% increased risk (95% CI 1 to 27%) of incident atrial fibrillation compared to those with the highest quintile. Dose-response spline analysis also showed an inverse association between mitochondria DNA copy number and hazard for atrial fibrillation for all three cohorts. These associations were consistent across subgroups. Mitochondria DNA copy number was inversely associated with the risk of AF independent of traditional cardiovascular risk factors. These findings implicate mitochondria DNA copy number as a novel risk factor for atrial fibrillation. Further research is warranted to understand the underlying mechanisms and to evaluate the role of mitochondria DNA copy number in the management of atrial fibrillation risk.


Reactivity studies of silylene [PhC(NtBu)2](C5Me5)Si - reactions with [M(COD)Cl]2 (M = Rh(i), Ir(i)), S, Se, Te, and BH3.
The reactivity of recently introduced N-heterocyclic silylene [PhC(NtBu)2](C5Me5)Si was evaluated. Three different reaction pathways were studied: (1) coordination to a Lewis acid, (2) oxidation by chalcogenides, and (3) coordination to transition metal complexes. The reaction of the silylene with BH3 led to the adduct [PhC(NtBu)2](C5Me5)Si(BH)3. Oxidation with the elemental chalcogens sulphur, selenium, and tellurium resulted in the corresponding silicon(iv)-chalcogenide compounds [PhC(NtBu)2](C5Me5)SiS, [PhC(NtBu)2](C5Me5)SiSe and [PhC(NtBu)2](C5Me5)SiTe. Treatment of the silylene with dimeric group 9 transition metal complexes [Ir(COD)Cl]2 and [Rh(COD)Cl]2 gave square planar coordinated transition metal silylene complexes [{PhC(NtBu)2}(C5Me5)Si-Rh(COD)(Cl)] and [{PhC(NtBu)2}(C5Me5)Si-Ir(COD)(Cl)].


The transmission and control of mastitis in dairy cows: a theoretical approach.
A multi-species model that incorporates the transmission of both major and minor mastitis pathogens as well as the interaction between them via coinfection of a quarter is fitted to data from seven dairy herds. The results suggest that major and minor pathogens can interact, on occasion, in a counter-intuitive way with implications for the control of clinical mastitis. The key finding is that delaying culling of cows with major pathogen infections for more than 100 days post infection could result in a higher prevalence of major pathogen infections, whereas early culling would reduce the levels. A theoretical exploration of current and proposed control strategies is carried out, informed by parameters estimated from the model and data. The results at each stage suggest of areas of further research such as: field-testing of the hypotheses presented; the exploration of a stochastic formulation of the model; analysis of the raw repeated measures data; application of control theory to determine the most effective combination of control strategies; inclusion of economic factors into the modelling framework.


Roles of fimB and fimE in site-specific DNA inversion associated with phase variation of type 1 fimbriae in Escherichia coli.
Evidence obtained with an improved in vivo assay of fimbrial phase variation in Escherichia coli supported a revised understanding of the roles of fimB and fimE in the site-specific DNA rearrangement with which they are associated. A previously proposed model argued that fimB and fimE play antagonistic, unidirectional roles in regulating the orientation of the invertible DNA element located immediately upstream of fimA, the gene encoding the major subunit of type 1 fimbriae. This conclusion, though, is based on an in vivo DNA inversion assay using recombinant plasmid substrates under conditions that, among other things, were incapable of detecting recombination of the fim invertible element from the on to the off orientation. Using a modified system that overcome this and several additional technical problems, we confirmed that fimB acts independently of fimE on the invertible element and that the additional presence of fimE results in the preferential rearrangement of the element to the off orientation. It is now demonstrated that fimE can act in the absence of fimB in this recombination to promote inversion primarily from on to off. In contrast to the previous studies, the effect of fimB on a substrate carrying the invertible element in the on orientation could be examined. It was found that fimB mediates DNA inversion from on to off, as well as from off to on, and that, contrary to prior interpretations, the fimB-associated inversion occurs with only minimal orientational preference to the on phase.


Drug-induced liver injury.
Drug-induced liver injury (DILI) remains an important disease in clinical practice. It is difficult to predict, diagnose and manage. Studies in the peer-reviewed literature in the last 2 years, focusing on the diagnosis, prediction and management of DILI will be reviewed. Antibiotics remain the most common drug causing DILI in the United States and Europe. Expert opinion may still be the better method of diagnosing DILI compared with an objective tool such as the Roussel-Uclaf Causality Assessment Method. Hepatitis E represents an alternative diagnosis to some cases of presumed drug hepatotoxicity. There is ongoing research into the genetics of the pathophysiology and susceptibility of DILI. A genome-wide association study confirmed the association between human leukocyte antigen (HLA) class II and susceptibility to coamoxiclav (amoxicillin-clavulanic acid) induced DILI. There is new information on the protective effect of HLA-DRB1*07 family of alleles. MicroRNAs are a potential marker of DILI. Keratin variants may predict outcome of acute liver failure. N-acetylcysteine may be protective against DILI while taking antituberculosis medication. Recent findings in the genetics of pathophysiology and susceptibility of DILI can help with predicting and avoiding DILI in clinical practice and provide the foundation for ongoing research.


Spnr, a murine RNA-binding protein that is localized to cytoplasmic microtubules.
Previous studies in transgenic mice have established the importance of the 3' untranslated region (UTR) of the spermatid-specific protamine-1 (Prm-1) mRNA in its translational control during male germ cell development. To clone genes that mediate the translational repression or activation of the Prm-1 mRNA, we screened cDNA expression libraries made with RNA from pachytene spermatocytes and round spermatids, with an RNA probe corresponding to the 3' UTR of Prm-1. We obtained six independent clones that encode Spnr, a spermatid perinuclear RNA-binding protein. Spnr is a 71-kD protein that contains two previously described RNA binding domains. The Spnr mRNA is expressed at high levels in the testis, ovary, and brain, and is present in multiple forms in those tissues. Immunolocalization of the Spnr protein within the testis shows that it is expressed exclusively in postmeiotic germ cells and that it is localized to the manchette, a spermatid-specific microtubular array. Although the Spnr protein is expressed too late to be directly involved in the translational repression of Prm-1 specifically, we suggest that the Spnr protein may be involved in other aspects of spermatid RNA metabolism, such as RNA transport or translational activation.


Hepatitis C virus sequence divergence preserves p7 viroporin structural and dynamic features.
The hepatitis C virus (HCV) viroporin p7 oligomerizes to form ion channels, which are required for the assembly and secretion of infectious viruses. The 63-amino acid p7 monomer has two putative transmembrane domains connected by a cytosolic loop, and has both N- and C- termini exposed to the endoplasmic reticulum (ER) lumen. NMR studies have indicated differences between p7 structures of distantly related HCV genotypes. A critical question is whether these differences arise from the high sequence variation between the different isolates and if so, how the divergent structures can support similar biological functions. Here, we present a side-by-side characterization of p7 derived from genotype 1b (isolate J4) in the detergent 6-cyclohexyl-1-hexylphosphocholine (Cyclofos-6) and p7 derived from genotype 5a (isolate EUH1480) in n-dodecylphosphocholine (DPC). The 5a isolate p7 in conditions previously associated with a disputed oligomeric form exhibits secondary structure, dynamics, and solvent accessibility broadly like those of the monomeric 1b isolate p7. The largest differences occur at the start of the second transmembrane domain, which is destabilized in the 5a isolate. The results show a broad consensus among the p7 variants that have been studied under a range of different conditions and indicate that distantly related HCVs preserve key features of structure and dynamics.


Effect of background case characteristics on decisions in the delivery room.
The authors investigated whether obstetricians make different decisions about a medical test case depending on the characteristics of background cases that preceded the test case. Five hypothetical cases were sent to 1247 obstetricians. The outcome of interest was the proportion of physicians who elect to perform a cesarean on a borderline test case, presented with 4 background cases. Participants were randomly assigned to 1 of 3 conditions: 1) pathological background, in which the test case was preceded by abnormal cases, typically requiring cesarean; 2) physiological background, where the test case was preceded by relatively uncomplicated cases, often suggesting a less invasive treatment; and 3) control, where the test case appeared first. A significantly higher proportion of respondents chose a cesarean when the test case was preceded by physiological cases (75.4%) than when it was preceded by pathological cases (52.2%). This tendency was observed among those actively and not actively involved in obstetrics and in physicians with different levels of training. A patient's chances of undergoing cesarean section can be influenced by the immediately prior experience of the physician. This study with hypothetical vignettes found that background cases can influence physicians' decisions. The test case was apparently perceived as more grave when it followed uncomplicated cases as compared to when it was preceded by abnormal cases. Such inconsistencies in decision making are unlikely to be fully resolved by expertise, as suggested by the lack of differences between physicians with different training levels. An understanding of such effects may contribute to more informed consideration of unappreciated influences in making decisions.


Involvement of DNA polymerase beta in repair of ionizing radiation damage as measured by in vitro plasmid assays.
Characteristic of damage introduced in DNA by ionizing radiation is the induction of a wide range of lesions. Single-strand breaks (SSBs) and base damages outnumber double-strand breaks (DSBs). If unrepaired, these lesions can lead to DSBs and increased mutagenesis. XRCC1 and DNA polymerase beta (polbeta) are thought to be critical elements in the repair of these SSBs and base damages. XRCC1-deficient cells display a radiosensitive phenotype, while proliferating polbeta-deficient cells are not more radiosensitive. We have recently shown that cells deficient in polbeta display increased radiosensitivity when confluent. In addition, cells expressing a dominant negative to polbeta have been found to be radiosensitized. Here we show that repair of radiation-induced lesions is inhibited in extracts with altered polbeta or XRCC1 status, as measured by an in vitro repair assay employing irradiated plasmid DNA. Extracts from XRCC1-deficient cells showed a dramatically reduced capacity to repair ionizing radiation-induced DNA damage. Extracts deficient in polbeta or containing a dominant negative to polbeta also showed reduced repair of radiation-induced SSBs. Irradiated repaired plasmid DNA showed increased incorporation of radioactive nucleotides, indicating use of an alternative long-patch repair pathway. These data show a deficiency in repair of ionizing radiation damage in extracts from cells deficient or altered in polbeta activity, implying that increased radiosensitivity resulted from radiation damage repair deficiencies.


10-undecynoic acid is a new anti-adherent agent killing biofilm of oral Streptococcus spp.
In the search for novel agents against oral pathogens in their planktonic and biofilm form, we have focused our attention on 10-undecynoic acid as the representative of the acetylenic fatty acids. Using macro-broth susceptibility testing method we first established MIC value. Next, the MBC value was determined from a broth dilution minimum inhibitory concentration test by sub-culturing it to BHI agar plates that did not contain the test agent. Anti-biofilm efficacy was tested in 96-well plates coated with saliva using BHI broth supplemented with 1% sucrose as a standard approach. Based on obtained results, MIC value for 10-undecynoic acid was established to be 2.5 mg/ml and the MBC value to be 5 mg/ml. The MBIC90 showed to be 2.5 mg/ml, however completed inhibition of biofilm formation was achieved at 5.0 mg/ml. MBBC concentration revealed to be the same as MBC value, causing approximately 30% reduction at the same time in biomass of pre-existing biofilm, whereas application of 7.0 mg/ml of 10-undecynoic acid crossed the 50% eradication mark. Strong anti-adherent effect was observed upon 10-undecynoic acid application at sub-MBC concentrations as well, complemented with suppression of acidogenicity and aciduricity. Thus, we concluded that 10-undecynoic acid might play an important role in the development of alternative or adjunctive antibacterial and anti-biofilm preventive and/or therapeutic approaches.


Interferometric sensing platform with dielectric nanostructured thin films.
A new interferometer-based optical sensing platform with nanostructured thin films of ZrO2 or TiO2 as sensing environment has been developed. With the application of an IC compatible Si(3)N(4) waveguide technology, Mach-Zehnder interferometer devices have been fabricated. The application of the glancing angle deposition technique allowed fabrication of nanostructured thin films as the optical sensing environment. Sensing ability of fabricated devices has been demonstrated through the refractive index measurement of a known gas. The transmission spectra and time response measurements have demonstrated a maximum phase shift of Delta phi=pi/10 and a |Delta P(out)|=0.65 dBm. Devices with TiO2 film on the sensing region performed much better than devices with ZrO2, with sensitivity twice as high.


Collection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae).
Salivary proteins of hematophagous Culicoides spp. are thought to play an important role in pathogen transmission and skin hypersensitivity. Analysis of these proteins, however, has been problematic due to the difficulty in obtaining adequate amounts of secreted Culicoides saliva. In the current study, a collection method for midge saliva was developed. Over a 3-d period, 3- to 5-d-old male and female Culicoides nubeculosus Meigen (Diptera: Ceratopogonidae) were repeatedly placed onto the collection system and allowed to deposit saliva into a filter. Salivary products were eluted from the filters and evaluated by gel electrophoresis and mass spectrometry as well as by intradermal testing and determination of clotting time. Gel electrophoresis revealed approximately 55 protein spots displaying relative molecular masses from 5 to 67 kDa and isoelectric points ranging from 4.5 to 9.8. The majority of molecular species analyzed by mass spectrometry showed high convergence with salivary proteins recently obtained from a cDNA library of Culicoides sonorensis Wirth & Jones, including proteins involved in sugarmeal digestion, defense, and coagulation inhibition as well as members of the D7 family and unclassified salivary proteins. In addition, the proteome analysis revealed a number of peptides that were related to proteins from insect species other than Culicoides. Intradermal injection of the saliva in human skin produced edema, vasodilatation, and pruritus. The anticoagulant activity of the saliva was demonstrated by significantly prolonged clotting times for human platelets. The potential role of the identified salivary proteins in the transmission of pathogens and the induction of allergies is discussed.


Mathematical biology and medical statistics: contributions to the understanding of AIDS epidemiology.
Some of the many ways in which mathematical biology and statistics have been used in investigating the acquired immunodeficiency syndrome (AIDS) epidemic are reviewed. Aspects of the spread of the disease via social and sexual networks are discussed. The different kinds of data involved are critically compared. Some studies of the incubation period are briefly reviewed and some comments made on the role of adherence to therapy.


Assessment of f-actin organization and apical-basal polarity during in vivo cat endothelial wound healing.
To assess the relationships between cytoskeletal changes and apical-basal polarity during healing of mechanical scrape injuries in the cat corneal endothelium. Ten cats (20 eyes) were used in this study. One mechanical scrape injury was created in the corneal endothelium of each eye using a blunt olive tip cannula. Tandem scanning confocal microscopy (TSCM) was performed at sequential time points after injury for in vivo assessment of cell morphology and wound healing rates. In two eyes, scanning electron microscopy was performed to allow verification of TSCM observations. Ten eyes were collected between 6 and 48 hours after wounding for in situ labeling of f-actin, ZO-1, or both. Cat endothelial cell morphology observed using in vivo microscopy was identical to that shown using scanning electron microscopy. During healing, endothelial cells always remained attached to the endothelial sheet, although some showed extensions of lamellipodia into the open wound area. The in situ localization of f-actin also correlated with the TSCM in vivo wound morphology. Quantitative analysis showed that there was a decrease in the intensity of phalloidin-fluorescein isothiocyanate staining at the leading edge of the wound, suggesting a decrease in f-actin; a significant correlation was found between the relative intensity of f-actin staining and the distance from the wound margin (R = 0.98, P < 0.01). At 24 and 48 hours after injury, both ZO-1 and f-actin maintained an apical localization within cells immediately adjacent to the leading edge, despite the considerable distance of movement and dramatic decrease in the intensity of f-actin staining. Overall, these data demonstrate that after scrape injury in the cat, endothelial cells exhibit a pattern of healing in which total intracellular f-actin is reduced, but normal cell connectivity and apical-basal polarity are maintained throughout.


Impaired growth and force production in skeletal muscles of young partially pancreatectomized rats: a model of adolescent type 1 diabetic myopathy?
This present study investigated the temporal effects of type 1 diabetes mellitus (T1DM) on adolescent skeletal muscle growth, morphology and contractile properties using a 90% partial pancreatecomy (Px) model of the disease. Four week-old male Sprague-Dawley rats were randomly assigned to Px (n = 25) or Sham (n = 24) surgery groups and euthanized at 4 or 8 weeks following an in situ assessment of muscle force production. Compared to Shams, Px were hyperglycemic (>15 mM) and displayed attenuated body mass gains by days 2 and 4, respectively (both P<0.05). Absolute maximal force production of the gastrocnemius plantaris soleus complex (GPS) was 30% and 50% lower in Px vs. Shams at 4 and 8 weeks, respectively (P<0.01). GP mass was 35% lower in Px vs Shams at 4 weeks (1.24±0.06 g vs. 1.93±0.03 g, P<0.05) and 45% lower at 8 weeks (1.57±0.12 vs. 2.80±0.06, P<0.05). GP fiber area was 15-20% lower in Px vs. Shams at 4 weeks in all fiber types. At 8 weeks, GP type I and II fiber areas were ∼25% and 40% less, respectively, in Px vs. Shams (group by fiber type interactions, P<0.05). Phosphorylation states of 4E-BP1 and S6K1 following leucine gavage increased 2.0- and 3.5-fold, respectively, in Shams but not in Px. Px rats also had impaired rates of muscle protein synthesis in the basal state and in response to gavage. Taken together, these data indicate that exposure of growing skeletal muscle to uncontrolled T1DM significantly impairs muscle growth and function largely as a result of impaired protein synthesis in type II fibers.


A comprehensive study comparing tack and glue mesh fixation in laparoscopic total extraperitoneal repair for adult groin hernias.
Glue mesh fixation is thought to cause less pain compared to tack mesh fixation during laparoscopic total extraperitoneal inguinal hernia repair (TEP). However, the clinical benefits of glue mesh fixation are still controversial. This study aimed to evaluate the acute pain, chronic pain, and recurrence rate between these two fixation methods. After reviewing all patients in our prospective hernia repair database from February 2008 to December 2017, we identified 583 patients who underwent TEP with tack mesh fixation and 70 patients with glue fixation by a single surgeon. Acute post-operative pain and activity level were evaluated using a Visual Analog Score (VAS) and the modified Medical Outcome Study (MOS) score. The primary endpoint was chronic pain 6 months after TEP. The secondary endpoints were acute pain, activity level, complications, and recurrence. After adjustment for potential confounding factors, the glue mesh fixation had significant lower VAS at 2 h post operation during rest and coughing and on the first day after surgery during coughing (p = 0.005, p < 0.001, and p = 0.011). The modified MOS on the first day was higher in the glue group (p < 0.001). There were no reduced risk of chronic pain or increased risk of recurrence for the glue group compared to the tack group [Odds ratio (OR) = 0.237, p = 0.169; OR = 2.498, p = 0.299]. In the sub-group analysis for recurrent hernia repair, glue fixation is associated with better modified MOS (p = 0.031) on first day and lower VAS on the operative day and first day at rest (p = 0.003 and p = 0.024) after surgery. Glue fixation method was superior to tack fixation method in acute post-operative pain and early post-operative activity level after laparoscopic TEP repair. However, both fixation methods had similar incidence of chronic pain-, recurrence-, and procedure-related complications after laparoscopic TEP repair.


Regulation of hippocampal synaptic plasticity thresholds and changes in exploratory and learning behavior in dominant negative NPR-B mutant rats.
The second messenger cyclic GMP affects synaptic transmission and modulates synaptic plasticity and certain types of learning and memory processes. The impact of the natriuretic peptide receptor B (NPR-B) and its ligand C-type natriuretic peptide (CNP), one of several cGMP producing signaling systems, on hippocampal synaptic plasticity and learning is, however, less well understood. We have previously shown that the NPR-B ligand CNP increases the magnitude of long-term depression (LTD) in hippocampal area CA1, while reducing the induction of long-term potentiation (LTP). We have extended this line of research to show that bidirectional plasticity is affected in the opposite way in rats expressing a dominant-negative mutant of NPR-B (NSE-NPR-BΔKC) lacking the intracellular guanylyl cyclase domain under control of a promoter for neuron-specific enolase. The brain cells of these transgenic rats express functional dimers of the NPR-B receptor containing the dominant-negative NPR-BΔKC mutant, and therefore show decreased CNP-stimulated cGMP-production in brain membranes. The NPR-B transgenic rats display enhanced LTP but reduced LTD in hippocampal slices. When the frequency-dependence of synaptic modification to afferent stimulation in the range of 1-100 Hz was assessed in transgenic rats, the threshold for both, LTP and LTD induction, was shifted to lower frequencies. In parallel, NPR-BΔKC rats exhibited an enhancement in exploratory and learning behavior. These results indicate that bidirectional plasticity and learning and memory mechanism are affected in transgenic rats expressing a dominant-negative mutant of NPR-B. Our data substantiate the hypothesis that NPR-B-dependent cGMP signaling has a modulatory role for synaptic information storage and learning.


Smoking re-initiation after cessation program: Comparison of associated factors between young and older adults.
To evaluate the associated factors in reinitiating the smoking habit among the participants of a smoking cessation program conducted in a tobacco cessation clinic of Manipal University, Manipal. This cross-sectional study was conducted among participants of a smoking cessation program who reinitiated smoking habit. A self-administered questionnaire was used that had information on demographic, habit history, knowledge on harmful effects of smoking behavior related to oral cavity and associated factors due to which individual was unable to quit the habit. A total of 102 males (mean age = 39.91 ± 9.57) constituted the final sample. The results showed that habitual smokers were more likely to be ≥40 years and occasional smokers were all reported to be <93 years (P < 0.001). Cigarette smokers were more likely to be of younger age group while majority of the Beedi and cigarette + Beedi smokers were older adults (P < 0.001). The mean duration of the habit was significantly higher for older adults than young adults (P < 0.001). There was no significant difference in the number of packs between the age groups (P = 0.054). A significantly higher proportion of young adults than older adults were aware about oral cancer (P < 0.001). Significantly higher proportion of older adults than young adults tend to have a closest person to be a smoker (P = 0.05). A significant higher proportion of young adults reason their habit as for pleasure (84.6%) and relaxation (68.8%), while older adults reason it to be as tension (64.1%) or combined factors (70.6%). Peer pressure was almost same in both the age groups (P = 0.006). There were no significant differences in the withdrawal symptoms among young and older adults (P = 0.41). Considerable differences were noticed between younger and older age groups in the factors which might play a role in re-intiating the smoking habit. A structured cessation program focused more on the above characteristics should be planned in public health programs based on the characteristics of the participants.