Generation and analysis of a mouse intestinal metatranscriptome through Illumina based RNA-sequencing.
With the advent of high through-put sequencing (HTS), the emerging science of metagenomics is transforming our understanding of the relationships of microbial communities with their environments. While metagenomics aims to catalogue the genes present in a sample through assessing which genes are actively expressed, metatranscriptomics can provide a mechanistic understanding of community inter-relationships. To achieve these goals, several challenges need to be addressed from sample preparation to sequence processing, statistical analysis and functional annotation. Here we use an inbred non-obese diabetic (NOD) mouse model in which germ-free animals were colonized with a defined mixture of eight commensal bacteria, to explore methods of RNA extraction and to develop a pipeline for the generation and analysis of metatranscriptomic data. Applying the Illumina HTS platform, we sequenced 12 NOD cecal samples prepared using multiple RNA-extraction protocols. The absence of a complete set of reference genomes necessitated a peptide-based search strategy. Up to 16% of sequence reads could be matched to a known bacterial gene. Phylogenetic analysis of the mapped ORFs revealed a distribution consistent with ribosomal RNA, the majority from Bacteroides or Clostridium species. To place these HTS data within a systems context, we mapped the relative abundance of corresponding Escherichia coli homologs onto metabolic and protein-protein interaction networks. These maps identified bacterial processes with components that were well-represented in the datasets. In summary this study highlights the potential of exploiting the economy of HTS platforms for metatranscriptomics.


Topical delivery of Avastin to the posterior segment of the eye in vivo using annexin A5-associated liposomes.
Effective delivery to the retina is presently one of the most challenging areas in drug development in ophthalmology, due to anatomical barriers preventing entry of therapeutic substances. Intraocular injection is presently the only route of administration for large protein therapeutics, including the anti-Vascular Endothelial Growth Factors Lucentis (ranibizumab) and Avastin (bevacizumab). Anti-VEGFs have revolutionised the management of age-related macular degeneration and have increasing indications for use as sight-saving therapies in diabetes and retinal vascular disease. Considerable resources have been allocated to develop non-invasive ocular drug delivery systems. It has been suggested that the anionic phospholipid binding protein annexin A5, may have a role in drug delivery. In the present study we demonstrate, using a combination of in vitro and in vivo assays, that the presence of annexin A5 can significantly enhance uptake and transcytosis of liposomal drug carrier systems across corneal epithelial barriers. This system is employed to deliver physiologically significant concentrations of Avastin to the posterior of the rat eye (127 ng/g) and rabbit retina (18 ng/g) after topical application. Our observations provide evidence to suggest annexin A5 mediated endocytosis can enhance the delivery of associated lipidic drug delivery vehicles across biological barriers, which may have therapeutic implications.


Prevalence of hyperprolactinemia in schizophrenia: association with typical and atypical antipsychotic treatment.
To evaluate the prevalence and severity of hyperprolactinemia among a large sample of patients with schizophrenia and related psychotic disorders treated with typical and atypical antipsychotic medications. Three electronic databases (general medical, psychiatric, and pharmacologic) containing the census data from November 2002 through March 2003 for a state-funded, inpatient hospital serving the chronically mentally ill were merged (N = 470). This database was purged of patient names, while the unique hospital identification number and demographic variables in each record were retained. These records were then screened to exclude patients with medications (except neuroleptics) or medical conditions known to elevate or suppress prolactin, leaving an overall sample (N = 422) in which to evaluate the prevalence of hyperprolactinemia. The sample was composed of patients with DSM-IV schizophrenia (N = 213), other related psychotic disorders (N = 131), mood disorders (N = 44), and other disorders (N = 34). For the overall sample (N = 422), which combined men and women, the mean serum prolactin level was 41.5 ng/mL; 290 of 422 patients were above the normal range. For women (N = 133), the mean serum prolactin level was 57.9 ng/mL, and 67% had levels above normal. For men (N = 289), the mean level was 33.9 ng/mL, with a 70% prevalence of hyperprolactinemia. While age did not influence the prevalence of elevated prolactin among men, age (reflecting reproductive status) was a significant variable in women; older age was associated with lower prolactin levels. For the study sample, a highly significant correlation was observed between neuroleptic dose (chlorpromazine equivalent) and serum prolactin level; however, this relationship was not determined on a medication-by-medication basis. Medications known to elevate prolactin were associated with higher prevalence rates of hyperprolactinemia, and "prolactin-sparing" medications had lower prevalence rates. However, when they were used in combination, the prolactin-elevating medication overwhelmed the effects of prolactin-sparing medication. This study suggested that neuroleptic treatment of schizophrenia is strongly associated with hyperprolactinemia and showed important differences between prolactin-sparing and prolactin-elevating medications.


On thin ice: barriers to adoption of surveillance for patients with stage I testicular seminoma. Survey of US radiation oncologists.
Most men with stage I testicular seminoma are cured with surgery alone, which is a preferred strategy per national guidelines. The current pattern of practice among US radiation oncologists (ROs) is unknown. We surveyed practicing US ROs via an online questionnaire. Respondent's characteristics, self-rated knowledge, perceived patient compliance rates with observation were analyzed for association with treatment recommendations. We received 353 responses from ROs, of whom 23% considered themselves experts. A vast majority (84%) recommend observation as a default strategy, however this rate drops to 3% if the patient is believed to be noncompliant. 33% of respondents believe that survival is jeopardized in case of disease recurrence, and among these respondents only 5% support observation. 22% of respondents over-estimate the likelihood of noncompliance with observation to be in the 50-80% range. Responders with a higher perceived noncompliance rate are more likely to recommend adjuvant therapy (Fisher's exact p<0.01). Only 7% of respondents recommend observation for stage IS seminoma and 45% administer adjuvant RT in patients with elevated pre-orchiectomy alpha-fetal protein levels. Many US ROs over-estimate the likelihood that stage I testicular seminoma patients will be noncompliant with surveillance and incorrectly believe that overall survival is jeopardized if disease recurs on surveillance. Observation is quickly dismissed for patients who are not deemed to be compliant with observation, and is generally not accepted for patients with stage IS disease. There is clearly an opportunity for improved physician education on evidence-based management of stage I testicular seminoma.


The influence of connective tissue growth factor on rabbit ligament injury repair.
The injured anterior cruciate ligament (ACL) is deemed to exhibit an impaired healing response and attempts at surgical repair have not been successful. Connective tissue growth factor (CTGF) is reported to be associated with wound healing, probably through transforming growth factor beta1 (TGF-β1). A rabbit ACL injury model was used to study the effect of CTGF on ligament recovery. Quantitative real-time PCR was performed for detection of changes in RNA levels of TGF-β1, type 1 collagen (COL-I), type 2 collagen (COL-II), SRY-related high mobility group-box gene9 (Sox9), metalloproteinase-1 (TIMP-1) as well as matrix metallopeptidase 13 (MMP-13). And expression of related proteins was detected by western blotting. The current study showed that CTGF could promote the recovery of inured anterior cruciate ligament. It can up-regulate the mRNA and expression of TGF-β1, COL-I, COL-II, Sox9, as well as the tissue inhibitor of TIMP-1, and down-regulated the mRNA and expression of MMP-13, suggesting the curative effect of CTGF on injured rabbit ligament is through regulating these cellular factors. This finding revealed the mechanism of CTFG's healing role in injured tissues and provided new possibilities of treating injured tissues and wound healing by using CTFG.


Cisapride treatment for gastro-oesophageal reflux in children.
Gastro-oesophageal reflux (GOR) is an extremely common and usually self-limiting condition in infants. When treatment is required, Cisapride, a pro-kinetic agent, has been commonly prescribed for the symptomatic management of GOR. There have been recent reports of possibly serious adverse events, e.g. an increased QTc interval, cardiac arrhythmias, and death, associated with the use of Cisapride. To determine the effectiveness of Cisapride for symptoms of GOR compared with placebo or any other non-surgical treatments. Searches were conducted of the Cochrane Central Trials Register and the specialised Trials register of the Cochrane Upper Gastrointestinal and Pancreatic Diseases Group, MEDLINE and Embase up till April 2002. Reference lists of relevant review articles and identified trials were scrutinised and forward citation searches were performed in the Science Citation Index on all trials identified. The search was re-run in August 2003 and no new trials were found. Randomised controlled trials that compared oral Cisapride therapy with placebo or with other non-surgical treatments for children with a diagnosis of GOR were included. Only studies in which Cisapride was administered orally for a minimum of one week and which documented at least one of the primary outcomes were included. We excluded trials in which the majority of participants were aged less than 28 days. The primary outcomes were defined as a change in symptoms at the end of treatment, presence of adverse events, occurrence of clinical complications, and weight gain. The secondary outcomes included physiological measures of GOR or histological evidence of oesophagitis. We dichotomised symptoms into 'same or worse' vs 'improved' and calculated summary odds ratios. Continuous measures of GOR (e.g. reflux index) were summarised as a weighted mean difference. All outcomes were analysed using a random effects method. Searches identified nine trials which met the inclusion criteria. Eight trials compared Cisapride with placebo, of which seven (236 participants) reported data on symptoms of gastro-oesophageal reflux, and one reported data on the QTc interval (49 patients). The odds ratio for 'same or worse' vs 'improved symptoms' at the end of treatment of 0.34 (95%CI 0.10, 1.19) did not show a statistically significant difference between the two interventions. There was significant heterogeneity between the studies and the funnel plot suggested publication bias. In a sensitivity analysis, the definition of outcomes was changed to 'any symptoms' vs 'no symptoms'. This resulted in the exclusion of three trials (one of them the largest, best quality trial). The resulting pooled odds ratio showed a significant effect of Cisapride (OR 0.19, 95%CI 0.08, 0.44). Five studies reported adverse events. Four reported adverse events (mainly diarrhoea) but the difference was not statistically significant (OR 1.80, 95%CI 0.87, 3.70). One trial found no difference in the QTc after 3 to 8 weeks of treatment. Cisapride was associated with a statistically significant reduction in the reflux index (weighted mean difference -6.49, 95%CI -10.13, -2.85), but as reflux index and clinical symptoms are poorly correlated, the clinical importance of this finding is uncertain. Other measures of oesophageal pH monitoring did not reach significance. One included study compared Cisapride with Gaviscon (or Gaviscon and Carobel). The odds ratio for 'same or worse' vs 'improvement' in the Cisapride group compared with Gaviscon was 3.26 (95%CI 0.93-11.38). We found no clear evidence that Cisapride reduces symptoms of GOR. The results suggested substantial publication bias favouring studies showing a positive effect of Cisapride. This finding is supported by the report of one unpublished multi-centre study of 134 patients, which was reported to show no evidence of a significant effect of Cisapride. Due to reports of fatal cardiac arrhythmias or sudden death, from July Due to reports of fatal cardiac arrhythmias or sudden death, from July 2000, cisapride was restricted to a limited access programme supervised by a paediatric gastrologist in the USA and in Europe, to patients treated within a clinical trial or safety study or registry programme.


β-Propeller protein-associated neurodegeneration: a new X-linked dominant disorder with brain iron accumulation.
Neurodegenerative disorders with high iron in the basal ganglia encompass an expanding collection of single gene disorders collectively known as neurodegeneration with brain iron accumulation. These disorders can largely be distinguished from one another by their associated clinical and neuroimaging features. The aim of this study was to define the phenotype that is associated with mutations in WDR45, a new causative gene for neurodegeneration with brain iron accumulation located on the X chromosome. The study subjects consisted of WDR45 mutation-positive individuals identified after screening a large international cohort of patients with idiopathic neurodegeneration with brain iron accumulation. Their records were reviewed, including longitudinal clinical, laboratory and imaging data. Twenty-three mutation-positive subjects were identified (20 females). The natural history of their disease was remarkably uniform: global developmental delay in childhood and further regression in early adulthood with progressive dystonia, parkinsonism and dementia. Common early comorbidities included seizures, spasticity and disordered sleep. The symptoms of parkinsonism improved with l-DOPA; however, nearly all patients experienced early motor fluctuations that quickly progressed to disabling dyskinesias, warranting discontinuation of l-DOPA. Brain magnetic resonance imaging showed iron in the substantia nigra and globus pallidus, with a 'halo' of T1 hyperintense signal in the substantia nigra. All patients harboured de novo mutations in WDR45, encoding a beta-propeller protein postulated to play a role in autophagy. Beta-propeller protein-associated neurodegeneration, the only X-linked disorder of neurodegeneration with brain iron accumulation, is associated with de novo mutations in WDR45 and is recognizable by a unique combination of clinical, natural history and neuroimaging features.


How health professionals facilitate parents' involvement in decision-making at the hospital: A parental perspective.
In many western countries, parents have a legal right to influence and be involved in decision-making (DM) surrounding their children's healthcare. This ensures that the healthcare is customized as far as possible to meet the children's and families' needs and preferences. However, parental involvement in such DM is not sufficiently implemented and the parental role during hospitalizations has become demanding. More knowledge is required to inform health professionals (HPs) about how to improve parental involvement in DM from a health-promoting perspective. The aims of this study were to explore parents' experiences of how HPs facilitate their involvement in the DM surrounding their child's healthcare and to identify how HPs can improve parental involvement at the hospital. This was an explorative, descriptive qualitative study within a constructivist research paradigm, comprising a purposive sample of 12 parents participating in individual semi-structured interviews. Qualitative content analysis was performed. The findings showed that HPs' sensitivity to parents' capacity, resources and needs was essential in order to facilitate the latter's involvement in DM. HPs' sensitivity also seemed to influence the quality of communication and the HP-parent relationship. Moreover, these factors appeared to affect parents' coping ability during their children's hospitalization.


Latanoprost could exacerbate the progression of presbyopia.
Prostaglandin analogues (PG) reduce intra-ocular pressure by enhancing uveoscleral flow at the ciliary body, which controls accommodation via the ciliary muscle. We investigated the effect of PG on accommodation and presbyopia progression in glaucoma patients. We conducted a clinic-based, retrospective, cross-sectional study. Inclusion criteria were bilateral phakic patients aged 40-69 years with best corrected visual acuity better than 20/30. Exclusion criteria were any disease affecting vision other than glaucoma and history of ocular surgery. Subjects with no prescription or vision-affecting disease served as controls (n = 260). The glaucoma patients were prescribed eye drops containing 0.005% latanoprost for more than six months (n = 23). We measured the binocular near add power at a distance of 30 cm in both groups and compared the results using Kaplan-Meier analysis. The mean age (± SD) of the control subjects was 51.5 ± 5.2 years and 39% were male. Similarly, the glaucoma patients had a mean age of 51.0 ± 7.2 years and 39% were male. There were no significant differences in age, gender, intra-ocular pressure, spherical equivalent, astigmatism, or anisometropia between groups. Survival analysis indicated that the glaucoma patients in this study reached the endpoint (near add power of +3.00 D) significantly earlier than control patients (P = 0.0001; generalized Wilcoxon test). Exacerbation of presbyopia progression in glaucoma patients is a potential side effect of latanoprost eyedrops.


Development of Streptomyces sp. FR-008 as an emerging chassis.
Microbial-derived natural products are important in both the pharmaceutical industry and academic research. As the metabolic potential of original producer especially Streptomyces is often limited by slow growth rate, complicated cultivation profile, and unfeasible genetic manipulation, so exploring a Streptomyces as a super industrial chassis is valuable and urgent. Streptomyces sp. FR-008 is a fast-growing microorganism and can also produce a considerable amount of macrolide candicidin via modular polyketide synthase. In this study, we evaluated Streptomyces sp. FR-008 as a potential industrial-production chassis. First, PacBio sequencing and transcriptome analyses indicated that the Streptomyces sp. FR-008 genome size is 7.26 Mb, which represents one of the smallest of currently sequenced Streptomyces genomes. In addition, we simplified the conjugation procedure without heat-shock and pre-germination treatments but with high conjugation efficiency, suggesting it is inherently capable of accepting heterologous DNA. In addition, a series of promoters selected from literatures was assessed based on GusA activity in Streptomyces sp. FR-008. Compared with the common used promoter ermE*-p, the strength of these promoters comprise a library with a constitutive range of 60-860%, thus providing the useful regulatory elements for future genetic engineering purpose. In order to minimum the genome, we also target deleted three endogenous polyketide synthase (PKS) gene clusters to generate a mutant LQ3. LQ3 is thus an "updated" version of Streptomyces sp. FR-008, producing fewer secondary metabolites profiles than Streptomyces sp. FR-008. We believe this work could facilitate further development of Streptomyces sp. FR-008 for use in biotechnological applications.


Detection of moniliformin in maize using capillary zone electrophoresis.
Moniliformin is a mycotoxin produced by certain fungi pathogenic to maize. It is capable of causing disease in domestic animals, possibly through inhibition of pyruvate dehydrogenase. Testing for MON commonly involves extraction of maize, isolation of moniliformin using solid-phase extraction columns and detection with high-performance liquid chromatography (HPLC) or gas chromatography. A capillary zone electrophoresis-diode array detection (CZE-DAD) method for determination of moniliformin in maize is reported. The extraction and isolation procedures are similar to those of a commonly used HPLC method, while the detection step requires only 10 min. Sixty-three samples of maize were tested by an established HPLC method using absorbance at 229 nm (HPLC-ultraviolet light) and by the CZE-DAD method. The limit of detection of the CZE-DAD method was 0.1 microg MON g(-1) maize compared with 0.05 microg g(-1) for the HPLC-ultraviolet light method. The CZE-DAD method gave good agreement with the HPLC-ultraviolet light method for samples tested at levels up to 1500 microg g(-1), with a linear regression of r(2) = 0.996.


Monocyte-derived macrophages exhibit distinct and more restricted HIV-1 integration site repertoire than CD4(+) T cells.
The host genetic landscape surrounding integrated HIV-1 has an impact on the fate of the provirus. Studies analysing HIV-1 integration sites in macrophages are scarce. We studied HIV-1 integration site patterns in monocyte-derived macrophages (MDMs) and activated CD4(+) T cells derived from seven antiretroviral therapy (ART)-treated HIV-1-infected individuals whose cells were infected ex vivo with autologous HIV-1 isolated during the acute phase of infection. A total of 1,484 unique HIV-1 integration sites were analysed. Their distribution in the human genome and genetic features, and the effects of HIV-1 integrase polymorphisms on the nucleotide selection specificity at these sites were indistinguishable between the two cell types, and among HIV-1 isolates. However, the repertoires of HIV-1-hosting gene clusters overlapped to a higher extent in MDMs than in CD4(+) T cells. The frequencies of HIV-1 integration events in genes encoding HIV-1-interacting proteins were also different between the two cell types. Lastly, HIV-1-hosting genes linked to clonal expansion of latently HIV-1-infected CD4(+) T cells were over-represented in gene hotspots identified in CD4(+) T cells but not in those identified in MDMs. Taken together, the repertoire of genes targeted by HIV-1 in MDMs is distinct from and more restricted than that of CD4(+) T cells.


Fingolimod protects against neonatal white matter damage and long-term cognitive deficits caused by hyperoxia.
Cerebral white matter injury is a leading cause of adverse neurodevelopmental outcome in prematurely born infants involving cognitive deficits in later life. Despite increasing knowledge about the pathophysiology of perinatal brain injury, therapeutic options are limited. In the adult demyelinating disease multiple sclerosis the sphingosine-1-phosphate (S1P) receptor modulating substance fingolimod (FTY720) has beneficial effects. Herein, we evaluated the neuroprotective potential of FTY720 in a neonatal model of oxygen-toxicity, which is associated with hypomyelination and impaired neuro-cognitive outcome. A single dose of FTY720 (1mg/kg) at the onset of neonatal hyperoxia (24h 80% oxygen on postnatal day 6) resulted in improvement of neuro-cognitive development persisting into adulthood. This was associated with reduced microstructural white matter abnormalities 4 months after the insult. In search of the underlying mechanisms potential non-classical (i.e. lymphocyte-independent) pathways were analysed shortly after the insult, comprising modulation of oxidative stress and local inflammatory responses as well as myelination, oligodendrocyte degeneration and maturation. Treatment with FTY720 reduced hyperoxia-induced oxidative stress, microglia activation and associated pro-inflammatory cytokine expression. In vivo and in vitro analyses further revealed that oxygen-induced hypomyelination is restored to control levels, which was accompanied by reduced oligodendrocyte degeneration and enhanced maturation. Furthermore, hyperoxia-induced elevation of S1P receptor 1 (S1P1) protein expression on in vitro cultured oligodendrocyte precursor cells was reduced by activated FTY720 and protection from degeneration is abrogated after selective S1P1 blockade. Finally, FTY720s' classical mode of action (i.e. retention of immune cells within peripheral lymphoid organs) was analysed demonstrating that FTY720 diminished circulating lymphocyte counts independent from hyperoxia. Cerebral immune cell counts remained unchanged by hyperoxia and by FTY720 treatment. Taken together, these results suggest that beneficial effects of FTY720 in neonatal oxygen-induced brain injury may be rather attributed to its anti-oxidative and anti-inflammatory capacity acting in concert with a direct protection of developing oligodendrocytes than to a modulation of peripheral lymphocyte trafficking. Thus, FTY720 might be a potential new therapeutic option for the treatment of neonatal brain injury through reduction of white matter damage.


Micropatterned biofilm formations by laminar flow-templating.
We present a microfluidic device capable of patterning linear biofilm formations using a flow templating approach. We describe the design considerations and fabrication methodology of a two level flow-templating micro-bioreactor (FT-μBR), which generates a biofilm growth stream surrounded on 3 sides by a growth inhibiting confinement stream. Through a combination of experiments and simulations we comprehensively evaluate and exploit control parameters to manipulate the biofilm growth template stream dimensions. The FT-μBR is then used to grow biofilm patterns with controllable dimensions. A proof-of-principle study using the device demonstrates its utility in conducting biofilm growth rate measurements under different shear stress environments. This opens the way for quantitative studies into the effects of the local shear environment on biofilm properties and for the synthesis of a new generation of functional biomaterials with controllable properties.


Application of comparative phylogenomics to study the evolution of Yersinia enterocolitica and to identify genetic differences relating to pathogenicity.
Yersinia enterocolitica, an important cause of human gastroenteritis generally caused by the consumption of livestock, has traditionally been categorized into three groups with respect to pathogenicity, i.e., nonpathogenic (biotype 1A), low pathogenicity (biotypes 2 to 5), and highly pathogenic (biotype 1B). However, genetic differences that explain variation in pathogenesis and whether different biotypes are associated with specific nonhuman hosts are largely unknown. In this study, we applied comparative phylogenomics (whole-genome comparisons of microbes with DNA microarrays combined with Bayesian phylogenies) to investigate a diverse collection of 94 strains of Y. enterocolitica consisting of 35 human, 35 pig, 15 sheep, and 9 cattle isolates from nonpathogenic, low-pathogenicity, and highly pathogenic biotypes. Analysis confirmed three distinct statistically supported clusters composed of a nonpathogenic clade, a low-pathogenicity clade, and a highly pathogenic clade. Genetic differences revealed 125 predicted coding sequences (CDSs) present in all highly pathogenic strains but absent from the other clades. These included several previously uncharacterized CDSs that may encode novel virulence determinants including a hemolysin, a metalloprotease, and a type III secretion effector protein. Additionally, 27 CDSs were identified which were present in all 47 low-pathogenicity strains and Y. enterocolitica 8081 but absent from all nonpathogenic 1A isolates. Analysis of the core gene set for Y. enterocolitica revealed that 20.8% of the genes were shared by all of the strains, confirming this species as highly heterogeneous, adding to the case for the existence of three subspecies of Y. enterocolitica. Further analysis revealed that Y. enterocolitica does not cluster according to source (host).


[Action-oriented information systems: subsidies for activities by workers' health programs at the local level].
Over the course of recent decades, Brazil has become an industrialized society. From an epidemiological perspective and considering changes that have been occurring in the Brazilian population, particularly concerning demographic and epidemiological transitions, this paper discusses the need to adapt the epidemiological surveillance system in order to incorporate the surveillance of non-transmissible diseases. The authors analyze the information system for work-related accidents in the Workers' Health Program, under SUDS (the Unified Decentralized Health System) for the Mandaqui Region (in the Northern part of the city of São Paulo), based on data from a local metalworking industry identified by the information system as a high-risk site for work-related accidents. The article also focuses on the importance of organized workers participating in actions to control work-related accidents. Although such participation is not always possible, the health sector must incorporate the workers' own concrete experience in order to develop more effective actions to prevent accidents at the workplace.


Association between FGF23, α-Klotho, and Cardiac Abnormalities among Patients with Various Chronic Kidney Disease Stages.
Several experimental studies have demonstrated that fibroblast growth factor 23 (FGF23) may induce myocardial hypertrophy via pathways independent of α-Klotho, its co-factor in the induction of phosphaturia. On the other hand, few studies have clearly demonstrated the relationship between FGF23 level and left ventricular hypertrophy among subjects without chronic kidney disease (CKD; i.e., CKD stage G1 or G2). To investigate the data from 903 patients admitted to the cardiology department with various degrees of renal function, including 234 patients with CKD stage G1/G2. Serum levels of full-length FGF23 and α-Klotho were determined by enzyme immunoassay. After adjustment for sex, age, and estimated glomerular filtration rate (eGFR), the highest FGF23 tertile was significantly associated with left ventricular hypertrophy among patients with CKD stage G1/G2 and those with CKD stage G3a/G3b/G4 as compared with the lowest FGF23 tertile, and the association retained significance after further adjustment for serum levels of corrected calcium, inorganic phosphate, and C-reactive protein, as well as diuretic use, history of hypertension, and systolic blood pressure. FGF23 was also associated with low left ventricular ejection fraction among patients with CKD stage G1/G2 and those with CKD stage G3a/G3b/G4 after adjusting for age, sex, eGFR, corrected calcium, and inorganic phosphate. On the other hand, compared with the highest α-Klotho tertile, the lowest α-Klotho tertile was associated with left ventricular hypertrophy and systolic dysfunction only among patients with CKD stage G3b and stage G3a, respectively. An association between FGF23 and cardiac hypertrophy and systolic dysfunction was observed among patients without CKD as well as those with CKD after multivariate adjustment. However, the association between α-Klotho and cardiac hypertrophy and systolic dysfunction was significant only among patients with CKD G3b and G3a, respectively.


Three-dimensional scaffold-free microtissues engineered for cardiac repair.
Cardiovascular diseases, including myocardial infarction (MI), persist as the leading cause of mortality and morbidity worldwide. The limited regenerative capacity of the myocardium presents significant challenges specifically for the treatment of MI and, subsequently, heart failure (HF). Traditional therapeutic approaches mainly rely on limiting the induced damage or the stress on the remaining viable myocardium through pharmacological regulation of remodeling mechanisms, rather than replacement or regeneration of the injured tissue. The emerging alternative regenerative medicine-based approaches have focused on restoring the damaged myocardial tissue with newly engineered functional and bioinspired tissue units. Cardiac regenerative medicine approaches can be broadly categorized into three groups: cell-based therapies, scaffold-based cardiac tissue engineering, and scaffold-free cardiac tissue engineering. Despite significant advancements, however, the clinical translation of these approaches has been critically hindered by two key obstacles for successful structural and functional replacement of the damaged myocardium, namely: poor engraftment of engineered tissue into the damaged cardiac muscle and weak electromechanical coupling of transplanted cells with the native tissue. To that end, the integration of micro- and nanoscale technologies along with recent advancements in stem cell technologies have opened new avenues for engineering of structurally mature and highly functional scaffold-based (SB-CMTs) and scaffold-free cardiac microtissues (SF-CMTs) with enhanced cellular organization and electromechanical coupling for the treatment of MI and HF. In this review article, we will present the state-of-the-art approaches and recent advancements in the engineering of SF-CMTs for myocardial repair.


Set theoretic compression with an application to image coding.
We show that the complete information that is available after an image has been encoded is not just an approximate quantized image version, but a whole set of consistent images that contains the original image by necessity. From this starting point, we develop a set of tools to design a new class of encoders for image compression, based on a set decomposition and recombination of the image features. As an initial validation, we show the results of an experiment where these tools are used to modify the encoding process of block discrete cosine transform (DCT) coding in order to yield less blocking artifacts.


Rapid Visible Light-Mediated Controlled Aqueous Polymerization with In Situ Monitoring.
We report a simple procedure for rapid, visible light-mediated, controlled radical polymerization in aqueous solutions. Based on the photoelectron transfer reversible addition-fragmentation chain transfer (PET-RAFT) polymerization, fast chain propagation at room temperature in water was achieved in the presence of reductant and without prior deoxygenation. A systematic study correlating irradiation intensity and polymerization kinetics, enabled by in situ nuclear magnetic resonance spectroscopy, provided optimized reaction conditions. The versatility of this procedure was demonstrated through a rapid triblock copolymer synthesis, and incorporation of water-labile activated esters for direct conjugation of hydrophilic small molecules and proteins. In addition, this technique boasts excellent temporal control and provides a wide range of macromolecular materials with controlled molecular weights and narrow molecular weight distributions.


Elastic membrane heterogeneity of living cells revealed by stiff nanoscale membrane domains.
Many approaches have been developed to characterize the heterogeneity of membranes in living cells. In this study, the elastic properties of specific membrane domains in living cells are characterized by atomic force microscopy. Our data reveal the existence of heterogeneous nanometric scale domains with specific biophysical properties. We focused on glycosylphosphatidylinositol (GPI)-anchored proteins, which play an important role in membrane trafficking and cell signaling under both physiological and pathological conditions and which are known to partition preferentially into cholesterol-rich microdomains. We demonstrate that these GPI-anchored proteins reside within domains that are stiffer than the surrounding membrane. In contrast, membrane domains containing the transferrin receptor, which does not associate with cholesterol-rich regions, manifest no such feature. The heightened stiffness of GPI domains is consistent with existing data relating to the specific condensation of lipids and the slow diffusion rates of lipids and proteins therein. Our quantitative data may forge the way to unveiling the links that exist between membrane stiffness, molecular diffusion, and signaling activation.


Network analysis and percolation transition in hydrogen bonded clusters: nitric acid and water extracted by tributyl phosphate.
Extraction of polar molecules by amphiphilic species results in a complex variety of clusters whose topologies and energetics control phase behavior and efficiency of liquid-liquid separations. A computational approach including quantum mechanical vibrational frequency calculations and molecular dynamics simulation with intermolecular network theory is used to provide a robust assessment of extractant and polar solute association through hydrogen bonding in the tributyl phosphate (TBP)/HNO3/H2O/dodecane system for the first time. The distribution of local topologies of the TBP/HNO3/H2O hydrogen bonded clusters is shown to be consistent with an equilibrium binding model. Mixed TBP/HNO3/H2O clusters are predicted that have not been previously observable in experiment due to limitations in scattering and spectroscopic resolution. Vibrational frequency calculations are compared with experimental data to validate the experimentally observed TBP-HNO3-HNO3 Chain structure. At high nitric acid and water loading, large hydrogen-bonded clusters of 20 to 80 polar solutes formed. The cluster sizes were found to be exponentially distributed, consistent with a constant average solute association free energy in that size range. Due to the deficit of hydrogen bond donors in the predominantly TBP/HNO3 organic phase, increased water concentrations lower the association free energy and enable growth of larger cluster sizes. For sufficiently high water concentrations, changes in the cluster size distribution are found to be consistent with the formation of a percolating cluster rather than reverse micelles, as has been commonly assumed for the occurrence of an extractant-rich third phase in metal-free solvent extraction systems. Moreover, the compositions of the large clusters leading to percolation agrees with the 1 : 3 TBP : HNO3 ratio reported in the experimental literature for TBP/HNO3/H2O third phases. More generally, the network analysis of cluster formation from atomic level interactions could allow for control of phase behavior in multi-component solutions of species with a variety of hydrogen bond types.


SENSITIVITY OF ANTARCTIC UROSPORA PENICILLIFORMIS (ULOTRICHALES, CHLOROPHYTA) TO ULTRAVIOLET RADIATION IS LIFE-STAGE DEPENDENT(1).
The sensitivity of different life stages of the eulittoral green alga Urospora penicilliformis (Roth) Aresch. to ultraviolet radiation (UVR) was examined in the laboratory. Gametophytic filaments and propagules (zoospores and gametes) released from filaments were separately exposed to different fluence of radiation treatments consisting of PAR (P = 400-700 nm), PAR + ultraviolet A (UVA) (PA, UVA = 320-400 nm), and PAR + UVA + ultraviolet B (UVB) (PAB, UVB = 280-320 nm). Photophysiological indices (ETRmax , Ek , and α) derived from rapid light curves were measured in controls, while photosynthetic efficiency and amount of DNA lesions in terms of cyclobutane pyrimidine dimers (CPDs) were measured after exposure to radiation treatments and after recovery in low PAR; pigments of propagules were quantified after exposure treatment only. The photosynthetic conversion efficiency (α) and photosynthetic capacity (rETRmax ) were higher in gametophytes compared with the propagules. The propagules were slightly more sensitive to UVB-induced DNA damage; however, both life stages of the eulittoral inhabiting turf alga were not severely affected by the negative impacts of UVR. Exposure to a maximum of 8 h UVR caused mild effects on the photochemical efficiency of PSII and induced minimal DNA lesions in both the gametophytes and propagules. Pigment concentrations were not significantly different between PAR-exposed and PAR + UVR-exposed propagules. Our data showed that U. penicilliformis from the Antarctic is rather insensitive to the applied UVR. This amphi-equatorial species possesses different protective mechanisms that can cope with high UVR in cold-temperate waters of both hemispheres and in polar regions under conditions of increasing UVR as a consequence of further reduction of stratospheric ozone.


The effect of the work environment on future sleep disturbances: a systematic review.
Workers often attribute poor sleep to factors at work. Despite the large number of workers with sleep disturbances, there is a lack of consensus on the relationship between the work environment and sleep. The purpose of this systematic review therefore was to conduct a comprehensive evaluation. To this end, we employed standardized methods to systematically locate, review, and tabulate the results of prospective or randomized studies of the impact of work factors on sleep disturbances. From the 7981 articles located in five databases, 24 fulfilled our inclusion criteria and formed the base of the review including meta-analyses of the effect sizes. Results showed that the psychosocial work variables of social support at work, control, and organizational justice were related to fewer sleep disturbances, while high work demands, job strain, bullying, and effort-reward imbalance were related to more future sleep disturbances. Moreover, working a steady shift was associated with disturbances while exiting shift work was associated with less disturbed sleep. We conclude that psychosocial work factors and the scheduling of work have an impact on sleep disturbances and this might be utilized in the clinic as well as for planning work environments. Future research needs to employ better methodology and focus on underlying mechanisms.


Health care utilization & costs for cystic fibrosis patients with pulmonary infections.
To examine patterns of health care utilization and costs among cystic fibrosis (CF) patients with pulmonary infections. Retrospective administrative claims database analysis. We used administrative claims data (including both medical and pharmacy claims) to examine health care utilization and costs among CF patients with pulmonary infections over one year. We conducted a subgroup analysis in which we examined selected outcome measures among patients with tobramycin for inhalation (TIS) prescriptions by the number of TIS prescriptions filled. Among 1,064 CF patients identified with pulmonary infections, 80% had at least one CF-related office visit, 34% had a CF-related hospital stay, and 95% filled at least one prescription over one year. Total annual CF-related health care costs averaged $29,000 plus $20,000 for prescription drugs. In the subgroup analysis, there was a trend towards longer lengths of stay and higher inpatient costs with fewer numbers of TIS prescriptions filled. CF patients with pulmonary infections have substantial levels of health care utilization and costs.


Isolation and Characterization of a 2-Oxoglutarate Dependent Dioxygenase Involved in the Second-to-Last Step in Vindoline Biosynthesis.
Young leaves from Catharanthus roseus plants contain the enzymes which convert the monoterpenoid indole alkaloid, tabersonine by three hydroxylations, two methylations, and one acetylation step to vindoline. A novel direct enzyme assay has been developed for a hydroxylase involved in vindoline biosynthesis, which catalyzes the C4-hydroxylation of 2,3-dihydro-3-hydroxy-N(1)-methyltabersonine to the 3,4-dihydroxy derivative. The enzyme showed an absolute requirement for 2-oxoglutarate and enzymatic activity was enhanced by ascorbate, establishing it as a 2-oxoglutarate-dependent dioxygenase (EC 1.14.11.-). The hydroxylase exhibited specificity for position 4 of various alkaloid substrates. The enzyme exhibited a pH optima between 7 and 8 and an apparent molecular weight of 45,000. The appearance of 4-hydroxylase activity was developmentally regulated and was shown to be inducible by light treatment of seedlings. Substrate specificity studies of this enzyme for indole alkaloid substrate suggested that hydroxylation at position 3 and N-methylation occur prior to hydroxylation at position 4. This is in agreement with previous studies which suggest that C4-hydroxylation is the second to last step in vindoline biosynthesis in Catharanthus roseus.


Primary Immune Deficiency Treatment Consortium (PIDTC) report.
The Primary Immune Deficiency Treatment Consortium (PIDTC) is a network of 33 centers in North America that study the treatment of rare and severe primary immunodeficiency diseases. Current protocols address the natural history of patients treated for severe combined immunodeficiency (SCID), Wiskott-Aldrich syndrome, and chronic granulomatous disease through retrospective, prospective, and cross-sectional studies. The PIDTC additionally seeks to encourage training of junior investigators, establish partnerships with European and other International colleagues, work with patient advocacy groups to promote community awareness, and conduct pilot demonstration projects. Future goals include the conduct of prospective treatment studies to determine optimal therapies for primary immunodeficiency diseases. To date, the PIDTC has funded 2 pilot projects: newborn screening for SCID in Navajo Native Americans and B-cell reconstitution in patients with SCID after hematopoietic stem cell transplantation. Ten junior investigators have received grant awards. The PIDTC Annual Scientific Workshop has brought together consortium members, outside speakers, patient advocacy groups, and young investigators and trainees to report progress of the protocols and discuss common interests and goals, including new scientific developments and future directions of clinical research. Here we report the progress of the PIDTC to date, highlights of the first 2 PIDTC workshops, and consideration of future consortium objectives.


Acute respiratory distress syndrome and acute lung injury.
Acute respiratory distress syndrome (ARDS) is a life threatening respiratory failure due to lung injury from a variety of precipitants. Pathologically ARDS is characterised by diffuse alveolar damage, alveolar capillary leakage, and protein rich pulmonary oedema leading to the clinical manifestation of poor lung compliance, severe hypoxaemia, and bilateral infiltrates on chest radiograph. Several aetiological factors associated with the development of ARDS are identified with sepsis, pneumonia, and trauma with multiple transfusions accounting for most cases. Despite the absence of a robust diagnostic definition, extensive epidemiological investigations suggest ARDS remains a significant health burden with substantial morbidity and mortality. Improvements in outcome following ARDS over the past decade are in part due to improved strategies of mechanical ventilation and advanced support of other failing organs. Optimal treatment involves judicious fluid management, protective lung ventilation with low tidal volumes and moderate positive end expiratory pressure, multi-organ support, and treatment where possible of the underlying cause. Moreover, advances in general supportive measures such as appropriate antimicrobial therapy, early enteral nutrition, prophylaxis against venous thromboembolism and gastrointestinal ulceration are likely contributory reasons for the improved outcomes. Although therapies such as corticosteroids, nitric oxide, prostacyclins, exogenous surfactants, ketoconazole and antioxidants have shown promising clinical effects in animal models, these have failed to translate positively in human studies. Most recently, clinical trials with β2 agonists aiding alveolar fluid clearance and immunonutrition with omega-3 fatty acids have also provided disappointing results. Despite these negative studies, mortality seems to be in decline due to advances in overall patient care. Future directions of research are likely to concentrate on identifying potential biomarkers or genetic markers to facilitate diagnosis, with phenotyping of patients to predict outcome and treatment response. Pharmacotherapies remain experimental and recent advances in the modulation of inflammation and novel cellular based therapies, such as mesenchymal stem cells, may reduce lung injury and facilitate repair.


Hydrostatic Pressure Regulates MicroRNA Expression Levels in Osteoarthritic Chondrocyte Cultures via the Wnt/β-Catenin Pathway.
Mechanical loading and hydrostatic pressure (HP) regulate chondrocytes' metabolism; however, how mechanical stimulation acts remain unclear. MicroRNAs (miRNAs) play an important role in cartilage homeostasis, mechanotransduction, and in the pathogenesis of osteoarthritis (OA). This study investigated the effects of a cyclic HP (1-5 MPa), in both normal and OA human chondrocytes, on the expression of miR-27a/b, miR-140, miR-146a/b, and miR-365, and of their target genes (MMP-13, ADAMTS-5, IGFBP-5, and HDAC-4). Furthermore, we assessed the possible involvement of Wnt/β-catenin pathway in response to HP. Chondrocytes were exposed to HP for 3h and the evaluations were performed immediately after pressurization, and following 12, 24, and 48 h. Total RNA was extracted and used for real-time PCR. β-catenin was detected by Western blotting analysis and immunofluorescence. In OA chondrocytes, HP induced a significant increase (p < 0.01) of the expression levels of miR-27a/b, miR-140, and miR-146a, and a significant reduction (p < 0.01) of miR-365 at all analyzed time points. MMP-13, ADAMTS-5, and HDAC-4 were significantly downregulated following HP, while no significant modification was found for IGFBP-5. β-catenin levels were significantly increased (p < 0.001) in OA chondrocytes at basal conditions and significantly reduced (p < 0.01) by HP. Pressurization did not cause any significant modification in normal cells. In conclusion, in OA chondrocytes, HP restores the expression levels of some miRNAs, downregulates MMP-13, ADAMTS-5, and HDAC-4, and modulates the Wnt/β-catenin pathway activation.


Divorce between theory and practice: the system of public health training in the United States.
Many analysts have complained about the severe disconnect between public health as it is taught in schools of public health and public health as it is practiced in health departments. At least in the United States, few faculty members teaching in schools of public health have ever worked in public health departments. By the same token, few of those working in public health departments have degrees from schools of public health; most receive on-the-job training. This history traces the roots of this disconnect or "divorce between theory and practice." It finds that the 1930s were the prime years of community-based public health education, when the pressure of the Depression and the funding newly made available from the federal government by New Deal legislation encouraged practical training programs linked to local communities and health departments. The "divorce" began in the post-war period as an unintended consequence of the system for funding medical education and research at a time of general unpopularity of public health during the McCarthy era. Schools of public health were generally ignored in the 1950s and they began to adapt the strategy that continues today, of using research grants, primarily from the National Institutes of Health, to grow their faculty and facilities.


Social factors and private benefits influence landholders' riverine restoration priorities in tropical Australia.
Private land conservation is an essential component of conservation that requires organizing both protection and restoration actions accordingly. Yet private land conservation programs are often formulated to generate public benefits, with inadequate consideration of costs or benefits to private landholders. Landholders' willingness to participate in conservation programs depends on a complex set of social factors, and the benefits they expect from participation. However, these two attributes are commonly evaluated independent of one another. We addressed this limitation through interviews aimed at determining landholders': 1) willingness to participate in restoration programs; 2) barriers to participation; 3) prioritization of proposed riverine restoration actions; 4) expected public or private benefits for undertaking proposed riverine restoration actions; and 5) most preferred incentive for undertaking proposed restoration actions on their land. Our results revealed four main findings. First, landholders stated that biases towards ecological rather than production outcomes, impractical programs, and government mistrust (structural factors) were the major barriers that prevented them from participating in riverine restoration on their land. Second, private benefits influenced landholders' willingness to engage riverine restoration. Third, 'a sense of stewardship and improved landscape aesthetics' (an internal factor) was the most commonly reported private benefit. Fourth, the most preferred incentives for high priority restoration actions were cash for on-ground works, extension and community recognition. We highlight the importance of designing private land conservation programs that align with landholders' priorities and deliver public benefits.


Facial skin mapping: from single point bio-instrumental evaluation to continuous visualization of skin hydration, barrier function, skin surface pH, and sebum in different ethnic skin types.
Dry skin is one of the most important concerns of consumers worldwide. Despite huge efforts over several decades, the personal care industry still does not offer a perfect solution to satisfy the unmet needs of consumers for moisturising treatments in different ethnic groups. The paucity of data for the underlying cellular and biochemical problems in, and the effects of moisturisers on photodamaged facial skin may partly explain this. Mainly, single point measurements are used to understand the effects of products on skin physiology even on surrogate skin sites such as the non-photodamaged volar forearm. Some groups have developed discontinuous facial maps of skin biophysical properties, however, in 2014 a continuous facial analysis of bio-instrumental evaluations was developed using a heat map approach. These maps enabled a continuous visualization of features that not only revealed an unexpected complexity of facial skin but also indicated that use of surrogate skin sites for facial skin is inappropriate. We have demonstrated that remarkable gradients of skin hydration, TEWL, skin surface pH and sebum exist within short distances across the face and the gradients are distinctive among different ethnic groups. In addition, these studies have demonstrated that darkly-pigmented individuals do not necessarily have a better skin barrier function than their less-pigmented counterparts and that Caucasians have a lower facial skin surface pH compared with more pigmented subjects. Overall, there are no correlations between capacitance, TEWL and skin surface pH including individual topology angle values. Novel 3D camera approaches have also been used to facilitate a more precise assignment of measurement sites and visualisation. The 3D facial colour mappings illustrated precisely the local moisturising effects of a moisturising cream. There were subtle ethnic differences in efficacy that may be related to underlying skin biochemistry and/or ethnic differences in product application. A placebo-controlled study using conductance measurements in Chinese subjects is also reported. Finally, a new whole face statistical approach has been taken to prove differences in skin parameters but also of moisturiser treatment that adds further to our understanding of the ethnic differences in skin physiology and product application. This paper reviews the background of the development and application of this methodology.


Recombinant vaccinia virus: immunization against multiple pathogens.
The coding sequences for the hepatitis B virus surface antigen, the herpes simplex virus glycoprotein D, and the influenza virus hemagglutinin were inserted into a single vaccinia virus genome. Rabbits inoculated intravenously or intradermally with this polyvalent vaccinia virus recombinant produced antibodies reactive to all three authentic foreign antigens. In addition, the feasibility of multiple rounds of vaccination with recombinant vaccinia virus was demonstrated.


Induction of thymidylate synthase as a 5-fluorouracil resistance mechanism.
Thymidylate synthase (TS) is a key enzyme in the de novo synthesis of 2'-deoxythymidine-5'-monophosphate (dTMP) from 2'-deoxyuridine-5'-monophosphate (dUMP), for which 5,10-methylene-tetrahydrofolate (CH(2)-THF) is the methyl donor. TS is an important target for chemotherapy; it is inhibited by folate and nucleotide analogs, such as by 5-fluoro-dUMP (FdUMP), the active metabolite of 5-fluorouracil (5FU). FdUMP forms a relatively stable ternary complex with TS and CH(2)THF, which is further stabilized by leucovorin (LV). 5FU treatment can induce TS expression, which might bypass dTMP depletion. An improved efficacy of 5FU might be achieved by increasing and prolonging TS inhibition, a prevention of dissociation of the ternary complex, and prevention of TS induction. In a panel of 17 colon cancer cells, including several variants with acquired resistance to 5FU, sensitivity was related to TS levels, but exclusion of the resistant variants abolished this relation. For antifolates, polyglutamylation was more important than the intrinsic TS level. Cells with low p53 levels were more sensitive to 5FU and the antifolate raltitrexed (RTX) than cells with high, mutated p53. Free TS protein down-regulates its own translation, but its transcription is regulated by E2F, a cell cycle checkpoint regulator. Together, this results in low TS levels in stationary phase cells. Although cells with a low TS might theoretically be more sensitive to 5FU, the low proliferation rate prevents induction of DNA damage and 5FU toxicity. TS levels were not related to polymorphisms of the TS promoter. Treatment with 5FU or RTX rapidly induced TS levels two- to five-fold. In animal models, 5FU treatment resulted in TS inhibition followed by a two- to three-fold TS induction. Both LV and a high dose of 5FU not only enhanced TS inhibition, but also prevented TS induction and increased the antitumor effect. In patients, TS levels as determined by enzyme activity assays, immunohistochemistry and mRNA expression, were related to a response to 5FU. 5FU treatment initially decreased TS levels, but this was followed by an induction, as seen with an increased ratio of TS protein over TS-mRNA. The clear retrospective relation between TS levels and response now forms the basis for a prospective study, in which TS levels are measured before treatment in order to determine the treatment protocol.


Global DNA methylation screening of liver in piperonyl butoxide-treated mice in a two-stage hepatocarcinogenesis model.
Disruptive epigenetic gene control has been shown to be involved in carcinogenesis. To identify key molecules in piperonyl butoxide (PBO)-induced hepatocarcinogenesis, we searched hypermethylated genes using CpG island (CGI) microarrays in non-neoplastic liver cells as a source of proliferative lesions at 25 weeks after tumor promotion with PBO using mice. We further performed methylation-specific polymerase chain reaction (PCR), real-time reverse transcription PCR, and immunohistochemical analysis in PBO-promoted liver tissues. Ebp4.1, Wdr6 and Cmtm6 increased methylation levels in the promoter region by PBO promotion, although Cmtm6 levels were statistically non-significant. These results suggest that PBO promotion may cause altered epigenetic gene regulation in non-neoplastic liver cells surrounding proliferative lesions to allow the facilitation of hepatocarcinogenesis. Both Wdr6 and Cmtm6 showed decreased expression in non-neoplastic liver cells in contrast to positive immunoreactivity in the majority of proliferative lesions produced by PBO promotion. These results suggest that both Wdr6 and Cmtm6 were spared from epigenetic gene modification in proliferative lesions by PBO promotion in contrast to the hypermethylation-mediated downregulation in surrounding liver cells. Considering the effective detection of proliferative lesions, these molecules could be used as detection markers of hepatocellular proliferative lesions and played an important role in hepatocarcinogenesis.


Dynamic construction of a neural network from multiple pattern generators in the lobster stomatogastric nervous system.
In the stomatogastric nervous system (STNS) of the lobster Homarus gammarus, the rhythmic discharge of a pair of identified modulatory neurons (PS cells) is able to construct de novo a functional network from neurons otherwise belonging to other functional networks. The PS interneurons are electrically coupled and possess endogenous oscillatory properties that can be activated synaptically by stimulation of an identified sensory pathway. PS neurons themselves project synaptically onto the three major neural networks (esophageal, gastric mill, and pyloric) of the STNS. When a PS is rhythmically active in vitro, either spontaneously (rarely) or in response to direct stimulation, it dramatically restructures the otherwise independent activity patterns of all three target networks. This functional reconfiguration elicited by a single cell does not rely on changes in neuronal allegiance to pre-existing circuits, or on a simple merger of these different circuits. Rather, PS is responsible for the creation of an entirely new motor rhythm in that, via its widespread synaptic connections, the interneuron is able to subjugate the ongoing activity of the three STNS circuits and selectively appropriate individual elements to its own intrinsic rhythm. In addition, PS excites motor neurons that innervate dilator muscles of a valve situated between the esophagus and the stomach. The reorganization of the regional foregut motor rhythms by the interneuron is therefore coordinated to the opening of this valve, which itself carries sensory receptors that have been found to activate bursting in PS. Our data suggest that the role of PS in massively restructuring stomatogastric output is to generate a unique motor pattern appropriate for swallowing-like behavior. In a wider context, moreover, the results demonstrate that a neural network may not exist as a predefined entity within the CNS, but may be dynamically assembled according to changing behavioral circumstances.


LIECHTENSTEIN VERSUS LAPAROSCOPIC TRANSABDOMINAL PREPERITONEAL (TAPP) HERNIA REPAIR: A PROSPECTIVE COMPARATIVE STUDY FOCUSED ON POSTOPERATIVE OUTCOMES IN A GENERAL SURGERY UNIT.
Three surgical techniques for inguinal hernia repair are currently validated. Few studies have compared results among Lichtenstein and transabdominal preperitoneal (TAPP) laparoscopic approach obtained at an early step of the learning curve. This study aims to compare the early treatment results between the Liechtenstein technique and the laparoscopic TAPP approach to provide a basis for the surgeon's decision-making. Patients were divided into two groups: those who underwent laparoscopic TAPP approach (114 patients), and those who underwent open Lichtenstein repair (35 patients). Data were collected from the medical records during the evolution of the immediate postoperative period and by telephone contact after hospital discharge. For the analysis of the variables, the chi-square test of independence was implemented, with a level of significance set at a p-value of 0.05. There was a strong association between laparoscopy, less postoperative pain, and longer operative time. In addition, a preference for the technique in cases of recurrence, bilaterality, associated umbilical hernia, or obesity was noticed. In this study, the Lichtenstein technique was associated with a shorter time to return to work and was the treatment of choice for elderly patients. TAPP laparoscopic herniorrhaphy should be the first choice in cases of bilaterality, associated umbilical hernia, obesity, and recurrence to a previous anterior repair. The surgical risk is adequate for the procedure, even at early stages of the learning curve.


Spatial analysis of land use impact on ground water nitrate concentrations.
In spatial analyses of causes or health effects of environmental pollutants, small units of analyses are usually preferred for internal environmental homogeneity reasons but can only be done when fine resolution data are available for most units. Objectives of this study were to determine which land use practices were spatially associated with ground water nitrate concentrations across Prince Edward Island (PEI), Canada, and which spatial aggregation is the preferred unit of analyses. Nitrate concentrations were determined for 4855 samples from private wells. Validated field-by-field land use data were available. Average nitrate concentration and percentage of area for the 14 major land use categories in PEI were determined for each of three spatial aggregations: watersheds based on topography and hydrology; freeform polygon boundaries based on similar neighboring nitrate concentrations; and 500-m buffer zones around each well. Results showed that the percentages of potato, grain, and hay coverage were positive predictors of ground water nitrate concentrations. Percentage of blueberry was a marginally significant negative predictor in the watershed and freeform polygon models, and percentage of residential coverage was a positive predictor in the freeform polygon and buffer zone models. Spatial autocorrelation was present in the freeform polygon and buffer zone models even after land use was taken into account. In conclusion, analyses based on watersheds produced the best predictive model with the percentages of land cover of potato, hay, and grain being significantly associated with ground water nitrate concentrations, and the percentages of blueberry, clear-cut woodland, and other agriculture being marginally significant.


The effect of report cards on the coverage of maternal and neonatal health care: a factorial, cluster-randomised controlled trial in Uttar Pradesh, India.
Report cards are a prominent strategy to increase the ability of citizens to express their view, improve public accountability, and foster community participation in the provision of health services in low-income and middle-income countries. In India, social accountability interventions that incorporate report cards and community meetings have been implemented at scale, attracting considerable policy attention, but there is little evidence on their effectiveness in improving health. We aimed to evaluate the effect of report cards, which contain information on village-level indicators of maternal and neonatal health care, and participatory meetings targeted at health providers and community members (including local leaders) on the coverage of maternal and neonatal health care in Uttar Pradesh, India. We conducted a repeated cross-sectional, 2 × 2 factorial, cluster-randomised controlled trial, in which each cluster was a village (rural) or ward (urban). The clusters were randomly assigned to one of four groups: the provider group, in which we shared report cards and held participatory meetings with providers of maternal and neonatal health services; the community group, in which we shared report cards and held participatory meetings with community members (including local leaders); the providers and community group, in which report cards were targeted at both health providers and the community; and the control group, in which report cards were not shared with anyone. We generated these report cards by collating data from household surveys and shared the report cards with the recipients (as determined by their assigned groups) in participatory meetings. The primary outcome was the proportion of women who had at least four antenatal care visits (ie, attended a clinic or were visited at home by a health-care worker) during their last pregnancy. We measured outcomes with cross-sectional household surveys that were taken at baseline, at a first follow-up (after 8 months of the intervention), and at a second follow-up (21 months after the start of the intervention). Analyses were by intention to treat. This trial is registered with ISRCTN, number ISRCTN11070792. We surveyed eligible women for the baseline survey between Jan 13, and Feb 5, 2015. We then randomly assigned 44 clusters to the provider group, 45 clusters to the community group, 45 clusters to the provider and community group, and 44 clusters to the control group. Report cards of collated survey data were provided to recipient groups, as per their random allocation, in October, 2015, and in September, 2016. We ran the first follow-up survey between May 16 and June 10, 2016. We ran the second follow-up survey between June 18 and July 18, 2017. We measured the primary outcome in 3133 women (795 in the provider group, 781 in the community group, 798 in the provider and community group, and 759 in the control group) who gave birth during implementation of the intervention, between Feb 1, 2016, and July 18, 2017 (the end of the second follow-up survey). The report card intervention did not significantly affect the proportion of women who had at least four antenatal care visits (provider vs non-provider: odds ratio 0·85, 95% CI 0·65-1·13; community vs non-community: 0·86, 0·65-1·13). Maternal health report cards containing information on village performance, targeted at either the community or health providers, had no detectable effect on the coverage of maternal and neonatal health care. Future research should seek to understand how the content of information and the delivery of report cards affect the success of this type of social accountability intervention. Merck Sharp and Dohme.


Isolation and structure of repressor-like proteins from the archaeon Sulfolobus solfataricus. Co-purification of RNase A with Sso7c.
The thermostable histone-like protein Sso7c (Sso for Sulfolobus solfataricus) from the archaeon Sulfolobus solfataricus was purified from the supernatant of acid-soluble cell lysates. Reverse phase HPLC of an apparently homogeneous Sso7c protein fraction from Mono S chromatography resulted in resolution of three further peaks. Sequence analysis revealed one of these components to be bovine RNase A, originating from the culture medium and explaining the RNA hydrolyzing activities of Sso7 preparations previously described. Sequence analysis of pure Sso7c showed an epsilon-Lys methylation pattern identical to that of Sso7d and a single Gln --> Glu mutational difference at position 13. The remaining two proteins obtained after HPLC separation were identified as homologues of bacterial repressor-like proteins. Thus, the existence of repressor-like proteins was demonstrated at the protein level in archaea, raising the question of structural and functional consequences of these proteins on the otherwise eukaryotic-like basal transcriptional machinery in archaea.


Integrating HIV services and other health services: A systematic review and meta-analysis.
Integration of HIV services with other health services has been proposed as an important strategy to boost the sustainability of the global HIV response. We conducted a systematic and comprehensive synthesis of the existing scientific evidence on the impact of service integration on the HIV care cascade, health outcomes, and cost-effectiveness. We reviewed the global quantitative empirical evidence on integration published between 1 January 2010 and 10 September 2021. We included experimental and observational studies that featured both an integration intervention and a comparator in our review. Of the 7,118 unique peer-reviewed English-language studies that our search algorithm identified, 114 met all of our selection criteria for data extraction. Most of the studies (90) were conducted in sub-Saharan Africa, primarily in East Africa (55) and Southern Africa (24). The most common forms of integration were (i) HIV testing and counselling added to non-HIV services and (ii) non-HIV services added to antiretroviral therapy (ART). The most commonly integrated non-HIV services were maternal and child healthcare, tuberculosis testing and treatment, primary healthcare, family planning, and sexual and reproductive health services. Values for HIV care cascade outcomes tended to be better in integrated services: uptake of HIV testing and counselling (pooled risk ratio [RR] across 37 studies: 1.67 [95% CI 1.41-1.99], p < 0.001), ART initiation coverage (pooled RR across 19 studies: 1.42 [95% CI 1.16-1.75], p = 0.002), time until ART initiation (pooled RR across 5 studies: 0.45 [95% CI 0.20-1.00], p = 0.050), retention in HIV care (pooled RR across 19 studies: 1.68 [95% CI 1.05-2.69], p = 0.031), and viral suppression (pooled RR across 9 studies: 1.19 [95% CI 1.03-1.37], p = 0.025). Also, treatment success for non-HIV-related diseases and conditions and the uptake of non-HIV services were commonly higher in integrated services. We did not find any significant differences for the following outcomes in our meta-analyses: HIV testing yield, ART adherence, HIV-free survival among infants, and HIV and non-HIV mortality. We could not conduct meta-analyses for several outcomes (HIV infections averted, costs, and cost-effectiveness), because our systematic review did not identify sufficient poolable studies. Study limitations included possible publication bias of studies with significant or favourable findings and comparatively weak evidence from some world regions and on integration of services for key populations in the HIV response. Integration of HIV services and other health services tends to improve health and health systems outcomes. Despite some scientific limitations, the global evidence shows that service integration can be a valuable strategy to boost the sustainability of the HIV response and contribute to the goal of 'ending AIDS by 2030', while simultaneously supporting progress towards universal health coverage.


Ultrasensitive Seawater pH Measurement by Capacitive Readout of Potentiometric Sensors.
Potentiometric pH probes remain the gold standard for the detection of pH but are not sufficiently sensitive to reliably detect ocean acidification at adequate frequency. Here, potentiometric probes are made dramatically more sensitive by placing a capacitive electronic component in series to the pH probe while imposing a constant potential over the measurement circuit. Each sample change now triggers a capacitive current transient that is easily identified between the two equilibrium states, and is integrated to reveal the accumulated charge. This affords dramatically higher precision than with traditional potentiometric probes. pH changes down to 0.001 pH units are easily distinguished in buffer and seawater samples, at a precision (standard deviation) of 28 μpH and 67 μpH, respectively, orders of magnitude better than what is possible with potentiometric pH probes.


Linac-based on-board imaging feasibility and the dosimetric consequences of head roll in head-and-neck IMRT plans.
Kilovoltage imaging systems on linear accelerators are used for patient localization in many clinics. The purpose of this work is to assess on-board imaging (OBI) detection of systematic setup errors and in particular, the dosimetric consequences of undetected head roll in head-and-neck intensity modulated radiation therapy (IMRT) plans when using these systems. The system used in this study was the Trilogy linear accelerator and associated software (Varian Medical Systems, Palo Alto, CA). Accuracy of OBI localization was evaluated using an anthropomorphic head phantom. The head phantom is rigidly attached to a specially designed positioning device with 5 degrees of freedom, 3 translational and 2 rotational in the axial and coronal planes. Simulated setup errors were 3 degrees and 5 degrees rotations in the axial plane and displacements of 5 mm in the left-right, anterior-posterior, and superior-inferior directions. The coordinates set by the positioning device were compared with the coordinates obtained as measured by using the image matching tools of paired 2-dimensional (2D) orthogonal image matching, and 3D cone-beam computed tomography (CT) volume matching. In addition, 6 physician-approved IMRT plans of nasopharynx and tonsil carcinoma were recalculated to evaluate the impact of undetected 3 degrees and 5 degrees head roll. Application of cone-beam CT (CBCT) for patient localization was superior to 2D matching techniques for detecting rotational setup errors. The use of CBCT allowed the determination of translational errors to within 0.5 mm, whereas kV planar was within 1 to 2 mm. Head roll in the axial plane was not easily detected with orthogonal image sets. Compared to the IMRT plans with no head roll, dose-volume histogram analysis demonstrated an average increase in the maximal spinal cord dose of 3.1% and 6.4% for 3 degrees and 5 degrees angles of rotation, respectively. Dose to the contralateral parotid was unchanged with 3 degrees roll and increased by 2.7% with 5 degrees roll. The results of this study show that volumetric setup verification using CBCT can improve bony anatomy setup detection to millimeter accuracy, and is a reliable method to detect head roll. However, the magnitude of possible dose errors due to undetected head roll suggests that CBCT does not need to be performed on a daily basis but rather weekly or bi-weekly to ensure fidelity of the head position with the immobilization system.


Increased thermogenic capacity of brown adipose tissue under low temperature and its contribution to arousal from hibernation in Syrian hamsters.
Brown adipose tissue (BAT) is thought to play a significant physiological role during arousal when body temperature rises from the extremely low body temperature that occurs during hibernation. The dominant pathway of BAT thermogenesis occurs through the β(3)-adrenergic receptor. In this study, we investigated the role of the β(3)-adrenergic system in BAT thermogenesis during arousal from hibernation both in vitro and in vivo. Syrian hamsters in the hibernation group contained BAT that was significantly greater in overall mass, total protein, and thermogenic uncoupling protein-1 than BAT from the warm-acclimated group. Although the ability of the β(3)-agonist CL316,243 to induce BAT thermogenesis at 36°C was no different between the hibernation and warm-acclimated groups, its maximum ratio over the basal value at 12°C in the hibernation group was significantly larger than that in the warm-acclimated group. Forskolin stimulation at 12°C produced equivalent BAT responses in these two groups. In vivo thermogenesis was assessed with the arousal time determined by the time course of BAT temperature or heart rate. Stimulation of BAT by CL316,243 significantly shortened the time of arousal from hibernation compared with that induced by vehicle alone, and it also induced arousal in deep hibernating animals. The β(3)-antagonist SR59230A inhibited arousal from hibernation either in part or completely. These results suggest that BAT in hibernating animals has potent thermogenic activity with a highly effective β(3)-receptor mechanism at lower temperatures.


Merlin deficiency predicts FAK inhibitor sensitivity: a synthetic lethal relationship.
The goal of targeted therapy is to match a selective drug with a genetic lesion that predicts for drug sensitivity. In a diverse panel of cancer cell lines, we found that the cells most sensitive to focal adhesion kinase (FAK) inhibition lack expression of the neurofibromatosis type 2 (NF2) tumor suppressor gene product, Merlin. Merlin expression is often lost in malignant pleural mesothelioma (MPM), an asbestos-induced aggressive cancer with limited treatment options. Our data demonstrate that low Merlin expression predicts for increased sensitivity of MPM cells to a FAK inhibitor, VS-4718, in vitro and in tumor xenograft models. Disruption of MPM cell-cell or cell-extracellular matrix (ECM) contacts with blocking antibodies suggests that weak cell-cell adhesions in Merlin-negative MPM cells underlie their greater dependence on cell-ECM-induced FAK signaling. This provides one explanation of why Merlin-negative cells are vulnerable to FAK inhibitor treatment. Furthermore, we validated aldehyde dehydrogenase as a marker of cancer stem cells (CSCs) in MPM, a cell population thought to mediate tumor relapse after chemotherapy. Whereas pemetrexed and cisplatin, standard-of-care agents for MPM, enrich for CSCs, FAK inhibitor treatment preferentially eliminates these cells. These preclinical results provide the rationale for a clinical trial in MPM patients using a FAK inhibitor as a single agent after first-line chemotherapy. With this design, the FAK inhibitor could potentially induce a more durable clinical response through reduction of CSCs along with a strong antitumor effect. Furthermore, our data suggest that patients with Merlin-negative tumors may especially benefit from FAK inhibitor treatment.


Electromyography Exposes Heterogeneity in Muscle Co-Contraction following Stroke.
Walking after stroke is often described as requiring excessive muscle co-contraction, yet, evidence that co-contraction is a ubiquitous motor control strategy for this population remains inconclusive. Co-contraction, the simultaneous activation of agonist and antagonist muscles, can be assessed with electromyography (EMG) but is often described qualitatively. Here, our goal is to determine if co-contraction is associated with gait impairments following stroke. Fifteen individuals with chronic stroke and nine healthy controls walked on an instrumented treadmill at self-selected speed. Surface EMGs were collected from the medial gastrocnemius (MG), soleus (SOL), and tibialis anterior (TA) of each leg. EMG envelope amplitudes were assessed in three ways: (1) no normalization, (2) normalization to the maximum value across the gait cycle, or (3) normalization to maximal M-wave. Three co-contraction indices were calculated across each agonist/antagonist muscle pair (MG/TA and SOL/TA) to assess the effect of using various metrics to quantify co-contraction. Two factor ANOVAs were used to compare effects of group and normalization for each metric. Co-contraction during the terminal stance (TSt) phase of gait is not different between healthy controls and the paretic leg of individuals post-stroke, regardless of the metric used to quantify co-contraction. Interestingly, co-contraction was similar between M-max and non-normalized EMG; however, normalization does not impact the ability to resolve group differences. While a modest correlation is revealed between the amount of TSt co-contraction and walking speed, the relationship is not sufficiently strong to motivate further exploration of a causal link between co-contraction and walking function after stroke. Co-contraction does not appear to be a common strategy employed by individuals after stroke. We recommend exploration of alternative EMG analysis approaches in an effort to learn more about the causal mechanisms of gait impairment following stroke.


Dopamine synthesis and dopamine receptor expression are disturbed in recurrent miscarriages.
l-dopa decarboxylase (DDC) is responsible for the synthesis of dopamine. Dopamine, which binds to the D2-dopamine receptor (D2R), plays an important role in the maintenance of pregnancy. Aim of our study was the analysis of DDC and D2R expression in placentas of spontaneous miscarriages (SMs) and recurrent miscarriages (RMs) in comparison to healthy controls. Patients with SM (n = 15) and RM (n = 15) were compared with patients from healthy pregnancies (n = 15) (pregnancy weeks 7-13 each). Placental tissue has been collected from SMs and RMs from the first trimester (Department of Gynaecology and Obstetrics, LMU Munich) and from abruptions (private practice, Munich). Placental cell lines, BeWo- and JEG-3 cells, were stimulated with the trace amines T0AM and T1AM in vitro. Levels of DDC and D2R in trophoblasts and the decidua were lower in RMs in comparison to healthy controls. Stimulation of BeWo cells with T1AM significantly reduced DDC mRNA and protein levels. Via double-immunofluorescence, a DDC-positive cell type beneath decidual stromal cells and foetal EVT in the decidua could be detected. Downregulation of DDC and D2R in trophoblasts of RMs reflects a reduced signal cascade of catecholamines on the foetal side.


Detecting kin selection at work using inclusive fitness.
A recent model shows that altruism can evolve with limited migration and variable group sizes, and the authors claim that kin selection cannot provide a sufficient explanation of their results. It is demonstrated, using a recent reformulation of Hamilton's original arguments, that the model falls squarely within the scope of inclusive fitness theory, which furthermore shows how to calculate inclusive fitness and the relevant relatedness. A distinction is drawn between inclusive fitness, which is a method of analysing social behaviour; and kin selection, a process that operates through genetic similarity brought about by common ancestry, but not by assortation by genotype or by direct assessment of genetic similarity. The recent model is analysed, and it turns out that kin selection provides a sufficient explanation to considerable quantitative accuracy, contrary to the authors' claims. A parallel analysis is possible and would be illuminating for all models of social behaviour in which individuals' effects on each other's offspring numbers combine additively.


ATM splicing variants as biomarkers for low dose dexamethasone treatment of A-T.
Ataxia Telangiectasia (AT) is a rare incurable genetic disease, caused by biallelic mutations in the Ataxia Telangiectasia-Mutated (ATM) gene. Treatment with glucocorticoid analogues has been shown to improve the neurological symptoms that characterize this syndrome. Nevertheless, the molecular mechanism underlying the glucocorticoid action in AT patients is not yet understood. Recently, we have demonstrated that Dexamethasone treatment may partly restore ATM activity in AT lymphoblastoid cells by a new ATM transcript, namely ATMdexa1. In the present study, the new ATMdexa1 transcript was also identified in vivo, specifically in the PMBCs of AT patients treated with intra-erythrocyte Dexamethasone (EryDex). In these patients it was also possible to isolate new "ATMdexa1 variants" originating from canonical and non-canonical splicing, each containing the coding sequence for the ATM kinase domain. The expression of the ATMdexa1 transcript family was directly related to treatment and higher expression levels of the transcript in patients' blood correlated with a positive response to Dexamethasone therapy. Neither untreated AT patients nor untreated healthy volunteers possessed detectable levels of the transcripts. ATMdexa1 transcript expression was found to be elevated 8 days after the drug infusion, while it decreased 21 days after treatment. For the first time, the expression of ATM splicing variants, similar to those previously observed in vitro, has been found in the PBMCs of patients treated with EryDex. These findings show a correlation between the expression of ATMdexa1 transcripts and the clinical response to low dose dexamethasone administration.


Subjective Cognitive Decline Is Associated With Altered Default Mode Network Connectivity in Individuals With a Family History of Alzheimer's Disease.
Both subjective cognitive decline (SCD) and a family history of Alzheimer's disease (AD) portend risk of brain abnormalities and progression to dementia. Posterior default mode network (pDMN) connectivity is altered early in the course of AD. It is unclear whether SCD predicts similar outcomes in cognitively normal individuals with a family history of AD. We studied 124 asymptomatic individuals with a family history of AD (age 64 ± 5 years). Participants were categorized as having SCD if they reported that their memory was becoming worse (SCD+). We used extensive neuropsychological assessment to investigate five different cognitive domain performances at baseline (n = 124) and 1 year later (n = 59). We assessed interconnectivity among three a priori defined ROIs: pDMN, anterior ventral DMN, medial temporal memory system (MTMS), and the connectivity of each with the rest of brain. Sixty-eight (55%) participants reported SCD. Baseline cognitive performance was comparable between groups (all false discovery rate-adjusted p values > .05). At follow-up, immediate and delayed memory improved across groups, but the improvement in immediate memory was reduced in SCD+ compared with SCD- (all false discovery rate-adjusted p values < .05). When compared with SCD-, SCD+ subjects showed increased pDMN-MTMS connectivity (false discovery rate-adjusted p < .05). Higher connectivity between the MTMS and the rest of the brain was associated with better baseline immediate memory, attention, and global cognition, whereas higher MTMS and pDMN-MTMS connectivity were associated with lower immediate memory over time (all false discovery rate-adjusted p values < .05). SCD in cognitively normal individuals is associated with diminished immediate memory practice effects and a brain connectivity pattern that mirrors early AD-related connectivity failure.


How I Do It: GreenLight XPS 180W photoselective vaporization of the prostate.
The treatments for benign prostate enlargement (BPE), also known as lower urinary tract symptoms secondary to benign prostatic hypertrophy (BPH-LUTS), have evolved significantly over recent years. Where transurethral resection of the prostate (TURP) has been the gold standard surgery for enlarged prostate glands < 80 grams, newer modalities such as laser technology have proliferated with safe and efficacious results. Notably, for prostates larger than 80-100 grams, the surgical options were an open, simple prostatectomy or perhaps a staged TURP. Both of these surgeries have the potential for bleeding complications, electrolyte abnormalities, and prolonged hospital admissions. Additional demographic and healthcare forces are also at play. Our aging population of men is being increasingly successfully treated for cardiovascular disease. This means more men are on anti-coagulation therapy, many of whom must stay on these drugs to prevent stent clotting or stroke. Hospital resources, especially overnight hospital admissions do add considerable strain to our healthcare systems. Men are also increasingly becoming more savvy consumers when it comes to their health. Many male patients would prefer to take as few medications as possible. Studies of BPH medications in Europe and the United States have shown drug discontinuation rates between 58%-70% at 1 year. Men who are faced with the choice of daily medication for life versus an outpatient procedure will often opt for the latter, which is in keeping with AUA guidelines that still put surgery as a patient choice alongside medications. Being able to offer GreenLight photoselective vaporization (GL-PVP) with the GreenLight XPS 180Watt system addresses all of these concerns. Men with bothersome BPH-LUTS with essentially any sized prostate gland, can be treated as same-day surgery requiring no overnight admission to hospital, while continuing necessary anti-coagulants, with significantly diminished risks of bleeding, erectile dysfunction, TUR-syndrome. Just as there are many ways to perform a TURP, techniques for GL-PVP do vary. The objectives of this article are to breakdown some of the basic steps for the novice user of GL-PVP, as well as impart some 'pearls' for the more experienced user. Nothing can replace hands-on experience for any surgery. The GL-PVP is unique in that there are guides such as this and previous articles, an excellent simulation device (GreenLight SIM), and mentoring programs in place. The success of many surgeries has been the standardization of the procedure. Performing GL-PVP should not be haphazard. A surgical plan based on prostate anatomy and size, cystoscopic appearance, and application of routinized techniques should yield consistent and optimal surgical outcomes.


A plant-derived wound therapeutic for cost-effective treatment of post-surgical scalp wounds with exposed bone.
To evaluate efficacy and cost-effectiveness of a plant-derived wound dressing (ONE), a mixture of hypericum oil (Hypericum perforatum L.) and neem oil (Azadirachta indica A. Juss.), in scalp wounds with exposed bone. All patients with scalp wounds with exposed bone, following the excision of skin tumors, and treated with ONE in 2011 were included. Time of healing, wound size, area of exposed bone, ease of handling, pain, and complications were evaluated. Costs of dressings and nursing time were compared with those cited in literature for other treatment modalities. Fifteen consecutive patients with a mean age of 76.87 ± 10.3 years (59-90 years) were analyzed. The mean wound size was 10.9 ± 6.84 cm(2) (0.4-22.6 cm(2)) with 4.8 ± 5.9 cm(2) (0.3-20.7 cm(2)) of exposed bone. The time of complete healing by secondary intention was 8.1 (4-20) weeks. Rapid formation of granulation tissue was observed which after 4 weeks covered the entire exposed bone surface in 11 of 15 cases (73%). Dressing change was simple with no pain reported; no infections or other complications occurred. Using ONE for a mean healing time of 56.7 days resulted in mean costs of EUR 423.73, which is substantially lower than those published for fascia lata, negative pressure therapy, or collagen matrix followed by skin grafting (EUR 1,612.82, EUR 4,411.80 and EUR 1,503.72, respectively). This retrospective, non-controlled analysis supports ONE as a simple-to-use and safe treatment option for scalp wounds with exposed bone. Treatment costs compare favorably to those published for other treatment modalities.


Early fragmentation in the adaptive voter model on directed networks.
We consider voter dynamics on a directed adaptive network with fixed out-degree distribution. A transition between an active phase and a fragmented phase is observed. This transition is similar to the undirected case if the networks are sufficiently dense and have a narrow out-degree distribution. However, if a significant number of nodes with low out degree is present, then fragmentation can occur even far below the estimated critical point due to the formation of self-stabilizing structures that nucleate fragmentation. This process may be relevant for fragmentation in current political opinion formation processes.


A photon turnstile dynamically regulated by one atom.
Beyond traditional nonlinear optics with large numbers of atoms and photons, qualitatively new phenomena arise in a quantum regime of strong interactions between single atoms and photons. By using a microscopic optical resonator, we achieved such interactions and demonstrated a robust, efficient mechanism for the regulated transport of photons one by one. With critical coupling of the input light, a single atom within the resonator dynamically controls the cavity output conditioned on the photon number at the input, thereby functioning as a photon turnstile. We verified the transformation from a Poissonian to a sub-Poissonian photon stream by photon counting measurements of the input and output fields. The results have applications in quantum information science, including for controlled interactions of single light quanta and for scalable quantum processing on atom chips.


Establishing a health-based recommended occupational exposure limit for nitrous oxide using experimental animal data - A systematic review protocol.
Nitrous oxide (N 2 O) is widely used as inhalation analgesic and anaesthetic in medical, paramedical, and veterinary practice. Previous evaluations resulted in classification of N 2 O as a possible risk factor for adverse reproductive health outcomes based on evidence from animal data. Available human data were considered inadequate, partly due to the possibility that other risk factors, such as co-exposures to other inhalation anaesthetics may have contributed to the adverse outcomes. As no substantial new human evidence has emerged since previous evaluations, this protocol describes a planned systematic review of the evidence obtained from animal studies. The aim is to assess the available evidence on the effects of N 2 O on reproductive and developmental outcomes in animals to inform a health-based recommended occupational exposure limit (OEL) for N 2 O. Comprehensive search strategies were designed to retrieve animal studies addressing N 2 O exposure from PubMed, EMBASE, and Web of Science. Screening of the studies retrieved will be performed by at least two independent reviewers, while discrepancies will be resolved by reaching consensus through repeated review and discussions. Articles will be included according to criteria specified in this protocol. Outcome data relevant for reproduction and development will be extracted and risk of bias will be assessed by two independent reviewers using the SYRCLE's risk of bias tool. Primary reproductive and developmental outcomes of interest will be the number of resorptions, malformations, and birth weight. We will focus on dose-response studies that allow to derive an OEL with the benchmark dose (BMD) approach. Adverse outcomes occurring at doses that are equivalent to the exposures occurring in human occupational settings will be particularly relevant for dose-response modelling. The proposed review has not been performed before. We will follow the procedures specified in this protocol. We will adhere to guidelines of Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), adapted for animal studies. Ethical approval will not be required, as the review will use existing data available in the public domain.