IL28B Genetic Variations in Patients with Recurrent Herpes Simplex Keratitis.
Background and objectives: Recurrent herpes simplex keratitis (HSK) is the most common cause of corneal blindness in the developed world. A relationship between host gene polymorphisms and the recurrence of herpes simplex virus (HSV) infection has previously been proposed. Thus, the aim of this study was to investigate a potential association between the IL28B host genotype and recurrent HSK. Materials and Methods: Eighty patients older than 18 years of age of both genders with a history of recurrent herpes simplex labialis (HSL) were considered for inclusion. Seventy-five of these patients were found to be seropositive for HSV-1 and were subsequently enrolled in the study. Twenty-four of the enrolled patients also had a history of recurrent HSK associated with severe corneal scarring and visual acuity deterioration. Total DNA was isolated from whole blood samples. A single-nucleotide polymorphism (SNP) rs12979860 near the IL28B gene on chromosome 19 was genotyped. Results: A significant association was observed between recurrent HSK and two SNPs of the IL28B genotype (CCrs12979860 and CTrs12979860, p < 0.01). The variation CCrs12979860 showed a significantly greater association with HSK (16 out of 26 patients) compared with CTrs12979860 (8 out of 34 patients). Conclusion: Seropositive individuals with a history of recurrent HSK are likely to have the CC IL28B genotype. This genotype may be related to incomplete control of the infection and more frequent periodical viral shedding along the first nerve branch of the trigeminal ganglion, which clinically manifests as recurrent herpes keratitis. The clinical manifestation of recurrent HSV-1 infection seems to be influenced by polymorphism of the IL28B genotype.


RNA Structure Refinement Using the ERRASER-Phenix Pipeline.
The final step of RNA crystallography involves the fitting of coordinates into electron density maps. The large number of backbone atoms in RNA presents a difficult and tedious challenge, particularly when experimental density is poor. The ERRASER-Phenix pipeline can improve an initial set of RNA coordinates automatically based on a physically realistic model of atomic-level RNA interactions. The pipeline couples diffraction-based refinement in Phenix with the Rosetta-based real-space refinement protocol ERRASER (Enumerative Real-Space Refinement ASsisted by Electron density under Rosetta). The combination of ERRASER and Phenix can improve the geometrical quality of RNA crystallographic models while maintaining or improving the fit to the diffraction data (as measured by R free). Here we present a complete tutorial for running ERRASER-Phenix through the Phenix GUI, from the command-line, and via an application in the Rosetta On-line Server that Includes Everyone (ROSIE).


Towards Predicting the Cytochrome P450 Modulation: From QSAR to Proteochemometric Modeling.
Drug metabolism determines the fate of a drug when it enters the human body and is a critical factor in defining their absorption, distribution, metabolism, excretion and toxicity (ADMET) characteristics. Among the various drug metabolizing enzymes, cytochrome P450s (CYP450) constitute an important protein family that aside from functioning in xenobiotic metabolism, is also responsible for a diverse array of other roles encompassing steroid and cholesterol biosynthesis, fatty acid metabolism, calcium homeostasis, neuroendocrine functions and growth regulation. Although CYP450 typically converts xenobiotics into safe metabolites, there are some situations whereby the metabolite is more toxic than its parent molecule. Computational modeling has been instrumental in CYP450 research by rationalizing the nature of the binding event (i.e. inhibit or induce CYP450s) or metabolic stability of query compounds of interest. A plethora of computational approaches encompassing ligand, structure and systems based approaches have been utilized to model CYP450-ligand interactions. This review provides a brief background on the CYP450 family (i.e. its roles, advantages and disadvantages as well as its modulators) and then discusses the various computational approaches that have been used to model CYP450-ligand interaction. Particular focus was given to the use of quantitative structure-activity relationship (QSAR) and more recent proteochemometric modeling studies. Finally, a perspective on the current state of the art and future trends of the field is also provided.


Correlating Function and Imaging Measures of the Medial Longitudinal Fasciculus.
To test the validity of diffusion tensor imaging (DTI) measures of tissue injury by examining such measures in a white matter structure with well-defined function, the medial longitudinal fasciculus (MLF). Injury to the MLF underlies internuclear ophthalmoparesis (INO). 40 MS patients with chronic INO and 15 healthy controls were examined under an IRB-approved protocol. Tissue integrity of the MLF was characterized by DTI parameters: longitudinal diffusivity (LD), transverse diffusivity (TD), mean diffusivity (MD) and fractional anisotropy (FA). Severity of INO was quantified by infrared oculography to measure versional disconjugacy index (VDI). LD was significantly lower in patients than in controls in the medulla-pons region of the MLF (p < 0.03). FA was also lower in patients in the same region (p < 0.0004). LD of the medulla-pons region correlated with VDI (R = -0.28, p < 0.05) as did FA in the midbrain section (R = 0.31, p < 0.02). This study demonstrates that DTI measures of brain tissue injury can detect injury to a functionally relevant white matter pathway, and that such measures correlate with clinically accepted evaluation indices for INO. The results validate DTI as a useful imaging measure of tissue integrity.


Thermal windows on Brazilian free-tailed bats facilitate thermoregulation during prolonged flight.
The Brazilian free-tailed bat (Tadarida brasiliensis) experiences challenging thermal conditions while roosting in hot caves, flying during warm daylight conditions, and foraging at cool high altitudes. Using thermal infrared cameras, we identified hot spots along the flanks of free-ranging Brazilian free-tailed bats, ventral to the extended wings. These hot spots are absent in syntopic cave myotis (Myotis velifer), a species that forages over relatively short distances, and does not engage in long-distance migration. We hypothesized that the hot spots, or "radiators," on Brazilian free-tailed bats may be adaptations for migration, particularly in this long-distance, high-flying species. We examined the vasculature of radiators on Brazilian free-tailed bats with transillumination to characterize the unique arrangements of arteries and veins that are positioned perpendicular to the body in the proximal region of the wing. We hypothesized that these radiators aid in maintaining heat balance by flushing the uninsulated thermal window with warm blood, thereby dissipating heat while bats are flying under warm conditions, but shunting blood away and conserving heat when they are flying in cooler air at high altitudes. We also examined fluid-preserved specimens representing 122 species from 15 of 18 chiropteran families and radiators appeared present only in species in the family Molossidae, including both sedentary and migratory species and subspecies. Thus, the radiator appears to be a unique trait that may facilitate energy balance and water balance during sustained dispersal, foraging, and long-distance migration.


Norepinephrine overflow in perfused mesenteric arteries of spontaneously hypertensive rats.
We examined the overflow of endogenous norepinephrine with electrical stimulation, the associated pressor response, and rate of initial neuronal uptake of [3H]norepinephrine in perfused mesenteric arteries of 7- and 13-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. The tissues of two rats, a spontaneously hypertensive and a WKY control rat, were simultaneously processed and subjected to the same electrical stimulation. Both absolute and fractional overflow of endogenous norepinephrine during periarterial nerve stimulation (5 and 10 Hz for 1 minute) in the tissue of 7-week-old SHR was significantly greater whereas overflow of 13-week-old SHR was equivalent as compared with that of the age-matched WKY rats. The tissue content of norepinephrine was 20-25% higher in SHR of both ages. There was significantly enhanced [3H]norepinephrine uptake in the tissues of young SHR, but no difference was observed in the older SHR. The pressor response to periarterial nerve stimulation was significantly enhanced in 7-week-old SHR and much more so at the older age as compared with the WKY control rats. Exogenous norepinephrine dose-response curves in the tissues of 7-week-old SHR exhibited a parallel leftward shift, characteristic of a change in sensitivity, whereas that of 13-week-old SHR showed a much steeper slope as compared with the respective WKY control rats. This finding suggests that in addition to smooth muscle supersensitivity, structural alterations had occurred in vasculature of 13-week-old SHR. These data indicate that in SHR both the exocytotic release of norepinephrine and the responsiveness of the vascular smooth muscle cells are enhanced in the developmental stage of hypertension whereas smooth muscle supersensitivity to norepinephrine and nonspecific structural alterations primarily contribute to the maintenance of hypertension at 13 weeks of age.


Infection with Leishmania (Leishmania) infantum of 0 to 18-Month-old children living in a visceral leishmaniasis-endemic area in Brazil.
The diagnosis of Leishmania (Leishmania) infantum infection in children from birth may serve as a reference for the early identification of cases that would progress to classical visceral leishmaniasis (VL) in endemic areas. This study prospectively evaluated newborns of mothers living in the municipality of Paracatu, Minas Gerais, Brazil. The infants were followed up at 6-month intervals by clinical examination, serological tests (immunofluorescence [IIF] and enzyme-linked immunosorbent assay with rK39 [ELISA-rK39]) and polymerase chain reaction (PCR) until they had completed 18 months of age. A total of 166 pregnant women were included to evaluate the possible transfer of antibodies or even congenital transmission. Twenty-two of the women tested positive by IIF, four by ELISA-rK39, and one by PCR. Three infants of the 25 women with some positive test results were also positive in the first test (one by IIF, one by ELISA-rK39, and the third by ELISA-rK39 and PCR). One hundred and sixty infants were included in the study; of these, 43 had at least one positive sample over time. However, agreement between tests was low. Follow-up of children with a positive result in the tests studied revealed no progression to classical disease within a period of 18 months. In contrast, two children with negative IIF, PCR, and ELISA-rK39 results developed classical VL at 9 and 12 months of age. In conclusion, a positive test result was variable and sometimes temporary and agreement between tests was low. Therefore, the early diagnosis of Leishmania infection was not associated with the early identification of cases that would progress to classical VL in the endemic area studied.


How to Recruit the Correct RNA Polymerase? Lessons from snRNA Genes.
Nuclear eukaryotic genomes are transcribed by three related RNA polymerases (Pol), which transcribe distinct gene sets. Specific Pol recruitment is achieved through selective core promoter recognition by basal transcription factors (TFs). Transcription by an inappropriate Pol appears to be rare and to generate mostly unstable products. A collection of short noncoding RNA genes [for example, small nuclear RNA (snRNA) or 7SK RNA genes], which play essential roles in processes such as maturation of RNA molecules or control of Pol II transcription elongation, possess highly similar core promoters, and yet are transcribed for some by Pol II and for others by Pol III as a result of small promoter differences. Here we discuss the mechanisms of selective Pol recruitment to such promoters.


Psychometric Validation of the English and French Versions of the Posttraumatic Stress Disorder Checklist for DSM-5 (PCL-5).
The purpose of this study is to assess the psychometric properties of a French version of the Posttraumatic Stress Disorder Checklist for DSM-5 (PCL-5), a self-report measure of posttraumatic stress disorder (PTSD) symptoms, and to further validate the existing English version of the measure. Undergraduate students (n = 838 English, n = 262 French) completed the PCL-5 as well as other self-report symptom measures of PTSD and depression online. Both the English and French versions PCL-5 total scores demonstrated excellent internal consistency (English: α = .95; French: α = .94), and strong convergent and divergent validity. Strong internal consistency was also observed for each of the four subscales for each version (α's > .79). Test-retest reliability for the French version of the measure was also very good (r = .89). Confirmatory factor analysis indicated that the four-factor DSM-5 model was not a good fit of the data. The seven-factor hybrid model best fit the data in each sample, but was only marginally superior to the six-factor anhedonia model. The French version of the PCL-5 demonstrated the same psychometric qualities as both the English version of the same measure and previous versions of the PCL. Thus clinicians serving French-speaking clients now have access to this highly used screening instrument. With regards to the structural validity of the PCL-5 and of the new PTSD diagnostic structure of the DSM-5, additional research is warranted. Replication of our results in clinical samples is much needed.


Acquired N-Linked Glycosylation Motifs in B-Cell Receptors of Primary Cutaneous B-Cell Lymphoma and the Normal B-Cell Repertoire.
Primary cutaneous follicle center lymphoma (PCFCL) is a rare mature B-cell lymphoma with an unknown etiology. PCFCL resembles follicular lymphoma (FL) by cytomorphologic and microarchitectural criteria. FL B cells are selected for N-linked glycosylation motifs in their B-cell receptors (BCRs) that are acquired during continuous somatic hypermutation. The stimulation of mannosylated BCR by lectins on the tumor microenvironment is therefore a candidate driver in FL pathogenesis. We investigated whether the same mechanism could play a role in PCFCL pathogenesis. Full-length functional variable, diversity, and joining gene sequences of 18 PCFCL and 8 primary cutaneous diffuse large B-cell lymphoma, leg-type were identified by unbiased Anchoring Reverse Transcription of Immunoglobulin Sequences and Amplification by Nested PCR and BCR reconstruction from RNA sequencing data. Low BCR variation demonstrated negligible ongoing somatic hypermutation in PCFCL and primary cutaneous diffuse large B-cell lymphoma, leg-type, and indicated that the PCFCL microarchitecture does not act as a functional germinal center. Similar to FL but in contrast to primary cutaneous diffuse large B-cell lymphoma, leg-type, BCR genes of 15 PCFCLs (83%) had acquired N-linked glycosylation motifs. These motifs were located at the BCR positions converted to N-linked glycosylation motifs in normal B-cell repertoires with low prevalence but mostly at different positions than those found in FL. The cutaneous localization of PCFCL might suggest a role for lectins from commensal skin bacteria in PCFCL lymphomagenesis.


Focus issue: systems analysis of protein phosphorylation.
Kinases and phosphatases are key regulatory molecules that participate in most cell signaling pathways. Systems-level analyses are providing new insights into phosphorylation sites and kinase specificity, and phosphoproteomic analyses are creating not just a wealth of data, but are also revealing rich revelations about cellular behavior, cellular responses to changing environmental conditions, and mechanisms of disease. Phosphatases are not merely signal terminators, but play active roles in signal transduction and are especially important in redox signaling. In this Focus Issue, Science Signaling highlights protein phosphorylation as the center of the signaling universe.


Development and application of an indirect competitive enzyme-linked immunosorbent assay for the detection of p,p'-DDE in human milk and comparison of the results against GC-ECD.
1,1-Dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p'-DDE) is the major metabolite of insecticide 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane (p,p'-DDT) and a persistent organic pollutant (POPs) with concerns regarding its bioaccumulation and persistence in the environment and food chain. In the present study, an indirect competitive enzyme-linked immunosorbant assay (ic-ELISA) specific for the detection of p,p'-DDE is described. In hapten synthesis, 2,2'-bis(4-chlorophenyl)ethanol and glutaric anhydride were used as precursor and spacer arm, respectively. The hapten was then conjugated to bovine serum albumin (BSA) as immunogen for mouse immunization and also conjugated to ovalbumin as coating antigen for ELISA. The developed ic-ELISA was used for detecting p,p'-DDE in human milk samples and validated against the results from conventional gas chromatography-electron capture detection (GC-ECD). Coefficients of variation (%CV) of ELISA were 5.7-10.4% for intra-assay and 10.6-19.6% for interassay variations. The Pearson correlation coefficient of p,p'-DDE concentrations between ic-ELISA and GC-ECD was r = 0.766, which was in an acceptable range. The results indicate that the developed assay could be an alternative analytical tool for monitoring p,p'-DDE in lipimic matrices such as human milk.


Identification of a Rice stripe necrosis virus resistance locus and yield component QTLs using Oryza sativa x O. glaberrima introgression lines.
Developing new population types based on interspecific introgressions has been suggested by several authors to facilitate the discovery of novel allelic sources for traits of agronomic importance. Chromosome segment substitution lines from interspecific crosses represent a powerful and useful genetic resource for QTL detection and breeding programs. We built a set of 64 chromosome segment substitution lines carrying contiguous chromosomal segments of African rice Oryza glaberrima MG12 (acc. IRGC103544) in the genetic background of Oryza sativa ssp. tropical japonica (cv. Caiapó). Well-distributed simple-sequence repeats markers were used to characterize the introgression events. Average size of the substituted chromosomal segments in the substitution lines was about 10 cM and covered the whole donor genome, except for small regions on chromosome 2 and 4. Proportions of recurrent and donor genome in the substitution lines were 87.59% and 7.64%, respectively. The remaining 4.78% corresponded to heterozygotes and missing data. Strong segregation distortion was found on chromosomes 3 and 6, indicating the presence of interspecific sterility genes. To illustrate the advantages and the power of quantitative trait loci (QTL) detection using substitution lines, a QTL detection was performed for scored traits. Transgressive segregation was observed for several traits measured in the population. Fourteen QTLs for plant height, tiller number per plant, panicle length, sterility percentage, 1000-grain weight and grain yield were located on chromosomes 1, 3, 4, 6 and 9. Furthermore, a highly significant QTL controlling resistance to the Rice stripe necrosis virus was located between SSR markers RM202-RM26406 (44.5-44.8 cM) on chromosome 11. Development and phenotyping of CSSL libraries with entire genome coverage represents a useful strategy for QTL discovery. Mapping of the RSNV locus represents the first identification of a genetic factor underlying resistance to this virus. This population is a powerful breeding tool. It also helps in overcoming hybrid sterility barriers between species of rice.


Endothelin receptor blockade prevents augmented extracellular matrix component mRNA expression and capillary basement membrane thickening in the retina of diabetic and galactose-fed rats.
Endothelins (ETs) are a family of vasoactive peptides that have mitogenic properties and have also been associated with altered long-term nuclear signaling. We have previously shown that mRNA levels for ET-1, ET-3, and their receptors are upregulated under hyperhexosemic conditions. In this study, an endothelin antagonist was used to assess the effects of endothelin blockage on the production of two basement membrane transcripts, fibronectin and collagen alpha1 (IV). The microvascular basement membranes were analyzed using ultrastructural morphometry. Streptozotocin-induced diabetic rats, galactose-fed rats (30% galactose in diet), and nondiabetic, non-galactose-fed control rats were studied after 1-month and 6-month follow-up. Simultaneously, similar animal groups were treated with a general ET receptor blocker (bosentan, 100 mg x kg(-1) x day(-1)) and investigated. Semiquantitative reverse transcription-polymerase chain reaction for fibronectin and collagen alpha1 (IV) was conducted after 1 month of follow-up with comparison to beta-actin housekeeping gene using slot blot hybridization and densitometry. Basement membrane thickness was assessed after 6 months of follow-up in diabetic rats, using the orthogonal intercept method. After 1 month of follow-up, increased fibronectin and collagen alpha1 (IV) mRNA were present in the retina of diabetic and galactosemic animals, and the bosentan-treated groups exhibited mRNA levels similar to the control animals. After 6 months of follow-up, diabetes and galactose feeding induced basement membrane thickening, which was partially prevented by bosentan treatment. The above findings indicate that increased production of extracellular matrix proteins leading to thickening of microvascular basement membrane, secondary to hyperhexosemia, may be mediated via augmented ET production.


Co-occurrence of personality disorders in persons with kleptomania: a preliminary investigation.
This study was conducted to examine the co-occurrence of personality disorders in a group of persons with kleptomania. Twenty-eight subjects with DSM-IV kleptomania were administered the Structured Clinical Interview for DSM-III-R Personality Disorders and a semistructured interview to assess demographics and clinical characteristics. Twelve subjects with kleptomania (42.9%) met criteria for at least one personality disorder. The most common were: paranoid (n = 5; 17.9%), schizoid (n = 3; 10.7%), and borderline (n = 3; 10.7%). Subjects with kleptomania combined with personality disorders had an earlier age of onset of stealing behavior (13.4 +/- 5.6 years compared with 27.4 +/- 14.2 years in those who had kleptomania only; t = 3.225; df = 26; p = .006). Severity of kleptomania symptoms did not differ among the Axis II comorbidities. Persons with kleptomania appear to have a high prevalence of personality disorders. Further studies are needed to understand the relationship of kleptomania to personality.


Event-related potential study of dynamic neural mechanisms of semantic organizational strategies in verbal learning.
Neuroimaging and neuropsychological data indicate that the frontal regions are implicated in semantic organizational strategies in verbal learning. Whereas these approaches tend to adopt a localizationist view, we used event-related potentials (ERPs) to investigate the dynamic neural mechanisms involved in these strategies. We recorded ERPs using a 128-channel system in 12 young adults (23.75+/-3.02 years) during 3 encoding conditions that manipulated the levels of semantic organization demands. In the Unrelated condition, the words to encode did not share any semantic attributes. For both Spontaneous and Guided conditions, the words in each list were drawn from four semantic categories. In the Spontaneous condition, participants were not informed about the semantic relationship between items. In contrast, in the Guided condition, participants were instructed to improve their subsequent recall by mentally regrouping related items with the aid of category labels. Results indicated that the P200 amplitude increased with the greater organizational demand of semantic strategies. In contrast, the late positive component (LPC) amplitude was larger in both encoding conditions with semantic related words regardless of their instructions as compared to the Unrelated condition. Finally, there was greater right frontal sustained activity in the Spontaneous condition than in the Unrelated condition. Thus, our data indicate that the P200 is sensitive to attentional processes that increase with the organizational semantic demand. The LPC indexes associative processes voluntarily involved in linking related items together. Finally, the right frontal region appears to play an important role in the self-initiation of semantic organizational strategies.


Enhanced ocular hypotensive response to epinephrine with prior dexamethasone treatment.
In 12 patients with ocular hypertension and similar pressures in each eye, one eye was randomly selected in a masked manner for pretreatment with three applications of topical 0.01% dexamethasone sodium phosphate drops prepared in artificial tears. The other eye was treated with three applications of drops of artificial tears alone. Both eyes were then treated with 0.1% epinephrine hydrochloride drops. Ocular pressures were measured for the next 2 hours at 20-minute intervals. When compared with eyes treated with epinephrine drops alone, eyes pretreated with dexamethasone drops showed a significantly greater decrease in pressure (mean [+/- SD] maximum difference, 3.1 +/- 2.1 mm Hg). Five patients had a difference of 1.0 to 4.0 mm Hg and five patients had a difference of 4.0 to 6.0 mm Hg. Two patients showed no response. Younger age was associated with the greater response. Adrenergic-glucocorticoid synergism resulting in the lowering of ocular pressure may offer a potential for therapeutic use in ocular hypertension and glaucoma.


Evaluating change in virtual reality adoption for brain injury rehabilitation following knowledge translation.
To evaluate the impact of knowledge translation (KT) on factors influencing virtual reality (VR) adoption and to identify support needs of therapists. Intervention will be associated with improvements in therapists' perceived ease of use and self-efficacy, and an associated increase in intentions to use VR. Single group mixed-methods pre-test-post-test evaluation of convenience sample of physical, occupational and rehabilitation therapists (n=37) from two brain injury rehabilitation centres. ADOPT-VR administered pre/post KT intervention, consisting of interactive education, clinical manual, technical and clinical support. Increases in perceived ease of use (p=0.000) and self-efficacy (p=0.001), but not behavioural intention to use VR (p=0.158) were found following KT, along with decreases in the frequency of perceived barriers. Post-test changes in the frequency and nature of perceived facilitators and barriers were evident, with increased emphasis on peer influence, organisational-level supports and client factors. Additional support needs were related to clinical reasoning, treatment programme development, technology selection and troubleshooting. KT strategies hold potential for targeting therapists' perceptions of low self-efficacy and ease of use of this technology. Changes in perceived barriers, facilitators and support needs at post-test demonstrated support for repeated evaluation and multi-phased training initiatives to address therapists' needs over time. Implications for Rehabilitation Therapists' learning and support needs in integrating virtual reality extend beyond technical proficiency to include clinical decision-making and application competencies spanning the entire rehabilitation process. Phased, multi-faceted strategies may be valuable in addressing therapists' changing needs as they progress from novice to experienced virtual reality users. The ADOPT-VR is a sensitive measure to re-evaluate the personal, social, environmental, technology-specific and system-level factors influencing virtual reality adoption over time.


Regulation of molecular chaperone gene transcription involves the serine phosphorylation, 14-3-3 epsilon binding, and cytoplasmic sequestration of heat shock factor 1.
Heat shock factor 1 (HSF1) regulates the transcription of molecular chaperone hsp genes. However, the cellular control mechanisms that regulate HSF1 activity are not well understood. In this study, we have demonstrated for the first time that human HSF1 binds to the essential cell signaling protein 14-3-3 epsilon. Binding of HSF1 to 14-3-3 epsilon occurs in cells in which extracellular signal regulated kinase (ERK) is activated and blockade of the ERK pathway by treatment with the specific ERK pathway inhibitor PD98059 in vivo strongly suppresses the binding. We previously showed that ERK1 phosphorylates HSF1 on serine 307 and leads to secondary phosphorylation by glycogen synthase kinase 3 (GSK3) on serine 303 within the regulatory domain and that these phosphorylation events repress HSF1. We show here that HSF1 binding to 14-3-3 epsilon requires HSF1 phosphorylation on serines 303 and 307. Furthermore, the serine phosphorylation-dependent binding of HSF1 to 14-3-3 epsilon results in the transcriptional repression of HSF1 and its sequestration in the cytoplasm. Leptomycin B, a specific inhibitor of nuclear export receptor CRM1, was found to reverse the cytoplasmic sequestration of HSF1 mediated by 14-3-3 epsilon, suggesting that CRM1/14-3-3 epsilon directed nuclear export plays a major role in repression of HSF1 by the ERK/GSK3/14-3-3 epsilon pathway. Our experiments indicate a novel pathway for HSF1 regulation and suggest a mechanism for suppression of its activity during cellular proliferation.


Effects of adsorbed and templated nanosilver in mesoporous calcium-silicate nanoparticles on inhibition of bacteria colonization of dentin.
Mesoporous calcium-silicate nanoparticles (MCSNs) are advanced biomaterials for controlled drug delivery and mineralization induction. Nanosilver-incorporated MCSNs (Ag-MCSNs) were prepared in the present study using both the adsorption and template methods. Both versions of Ag-MCSNs showed characteristic morphology of mesoporous materials and exhibited sustained release of ions over time. In antibacterial testing against planktonic Enterococcus faecalis, Ag-MCSNs showed significantly better antibacterial effects when compared with MCSNs (P<0.05). The Ag-MCSNs aggregated on the dentin surface of root canal walls and infiltrated into dentinal tubules after ultrasound activation, significantly inhibiting the adherence and colonization of E. faecalis on dentin (P<0.05). Despite this, Ag-MCSNs with templated nanosilver showed much lower cytotoxicity than Ag-MCSNs with adsorbed nanosilver (P<0.05). The results of the present study indicated that nanosilver could be incorporated into MCSNs using the template method. The templated nanosilver could release silver ions and inhibit the growth and colonization of E. faecalis both in the planktonic form and as biofilms on dentin surfaces as absorbed nanosilver. Templated Ag-MCSNs may be developed into a new intracanal disinfectant for root canal disinfection due to their antibacterial ability and low cytotoxicity, and as controlled release devices for other bioactive molecules to produce multifunctional biomaterials.


Characterization of protein complexes using targeted proteomics.
Biological systems are not only controlled by the abundance of individual proteins, but also by the formation of complexes and the dynamics of protein-protein interactions. The identification of the components of protein complexes can be obtained by shotgun proteomics using affinity purification coupled to mass spectrometry. Such studies include the analyses of several samples and experimental controls in order to discriminate true specific interactions from unspecific interactions and contaminants. However, shotgun proteomics have limited quantification capabilities for low abundant proteins on large sample sets due to the undersampling and the stochastic precursor ion selection. In this context, targeted proteomics constitutes a powerful analytical tool to systematically detect and quantify peptides in multiple samples, for instance those obtained from affinity purification experiments. Hypothesis-driven strategies have mainly relied on the selected reaction monitoring (SRM) technique performed on triple quadrupole instruments, which enables highly selective and sensitive measurements of peptides, acting as surrogates of the pre-selected proteins, over a wide range of concentrations. More recently, novel quantitative methods based on high resolution instruments, such as the parallel reaction monitoring (PRM) technique implemented on the quadrupole-orbitrap instrument, have arisen and provided alternatives to perform quantitative analyses with enhanced selectivity.The application of targeted proteomics to protein-protein interaction experiments from plasma and other physiological fluid samples and the inclusion of parallel reaction monitoring (PRM), combined with other recent technology developments opens a vast area for clinical application of proteomics. It is anticipated that it will reveal valuable information about specific, individual, responses against drugs, exogenous proteins or pathogens.


Cooperative nanomaterial system to sensitize, target, and treat tumors.
A significant barrier to the clinical translation of systemically administered therapeutic nanoparticles is their tendency to be removed from circulation by the mononuclear phagocyte system. The addition of a targeting ligand that selectively interacts with cancer cells can improve the therapeutic efficacy of nanomaterials, although these systems have met with only limited success. Here, we present a cooperative nanosystem consisting of two discrete nanomaterials. The first component is gold nanorod (NR) "activators" that populate the porous tumor vessels and act as photothermal antennas to specify tumor heating via remote near-infrared laser irradiation. We find that local tumor heating accelerates the recruitment of the second component: a targeted nanoparticle consisting of either magnetic nanoworms (NW) or doxorubicin-loaded liposomes (LP). The targeting species employed in this work is a cyclic nine-amino acid peptide LyP-1 (Cys-Gly-Asn-Lys-Arg-Thr-Arg-Gly-Cys) that binds to the stress-related protein, p32, which we find to be upregulated on the surface of tumor-associated cells upon thermal treatment. Mice containing xenografted MDA-MB-435 tumors that are treated with the combined NR/LyP-1LP therapeutic system display significant reductions in tumor volume compared with individual nanoparticles or untargeted cooperative system.


The selectivity of beta-adrenoceptor antagonists at the human beta1, beta2 and beta3 adrenoceptors.
Beta-adrenoceptor antagonists ("beta-blockers") are one of the most widely used classes of drugs in cardiovascular medicine (hypertension, ischaemic heart disease and increasingly in heart failure) as well as in the management of anxiety, migraine and glaucoma. Where known, the mode of action in cardiovascular disease is from antagonism of endogenous catecholamine responses in the heart (mainly at beta1-adrenoceptors), while the worrisome side effects of bronchospasm result from airway beta2-adrenoceptor blockade. The aim of this study was to determine the selectivity of beta-antagonists for the human beta-adrenoceptor subtypes. (3)H-CGP 12177 whole cell-binding studies were undertaken in CHO cell lines stably expressing either the human beta1-, beta2- or the beta3-adrenoceptor in order to determine the affinity of ligands for each receptor subtype in the same cell background. In this study, the selectivity of well-known subtype-selective ligands was clearly demonstrated: thus, the selective beta1 antagonist CGP 20712A was 501-fold selective over beta2 and 4169-fold selective over beta3; the beta2-selective antagonist ICI 118551 was 550- and 661-fold selective over beta1 and beta3, respectively, and the selective beta3 compound CL 316243 was 10-fold selective over beta2 and more than 129-fold selective over beta1. Those beta2-adrenoceptor agonists used clinically for the treatment of asthma and COPD were beta2 selective: 29-, 61- and 2818-fold for salbutamol, terbutaline and salmeterol over beta1, respectively. There was little difference in the affinity of these ligands between beta1 and beta3 adrenoceptors. The clinically used beta-antagonists studied ranged from bisoprolol (14-fold beta1-selective) to timolol (26-fold beta2-selective). However, the majority showed little selectivity for the beta1- over the beta2-adrenoceptor, with many actually being more beta2-selective. This study shows that the beta1/beta2 selectivity of most clinically used beta-blockers is poor in intact cells, and that some compounds that are traditionally classed as "beta1-selective" actually have higher affinity for the beta2-adrenoceptor. There is therefore considerable potential for developing more selective beta-antagonists for clinical use and thereby reducing the side-effect profile of beta-blockers.


Negative and positive cis-acting elements control the expression of murine alpha 1-protease inhibitor genes.
The alpha 1-protease inhibitor (alpha 1-PI) proteins of mice are encoded by a group of genes whose members are expressed coordinately in a liver-abundant pattern and are regulated primarily at the transcriptional level. To better understand the developmental and tissue-specific regulation of this gene family, one member that is analogous to the human alpha 1-antitrypsin gene was chosen for study. Deletional analysis of the upstream regulatory region of this gene was performed, spanning from -10 kilobases to -80 base pairs relative to the transcriptional start site. Two functional positive cis-acting elements within the 522 bases immediately upstream of the start site for transcription were shown to modulate the level of expression from this promoter when introduced into human or mouse hepatoma cells, and a third region acted as a negative regulatory element in that its deletion resulted in a two- to sixfold increase of expression of a transfected minigene construct. Sequence comparison between the regulatory domains of two mouse alpha 1-PI genes and the human alpha 1-antitrypsin gene showed that the mouse gene contains a novel positive cis-acting element which is absent in human gene and that a specific eight-base-pair difference between species results in a strong positive cis-acting element in the human gene acting as a negative element in the mouse gene. An enhancer located approximately 3,000 base pairs upstream of the major start site for transcription was also identified. This element is position and orientation independent. Several different DNA-protein binding assays were used to demonstrate that each DNA segment with functional significance in transfection assays interacts specifically with proteins found in adult mouse liver nuclei. The major positive-acting element appeared to be specifically recognized by nuclear proteins found only in tissues that express alpha 1-PI, while the negative element binding proteins were ubiquitous. Thus, the distal regulatory domain including bases -3500 to -133 of this murine alpha 1-PI gene family member is more complex than was previously demonstrated. It is composed of a set of at least three additional functional cis-acting regulatory elements besides those which have been mapped by others and has a far upstream enhancer.


Gastric Cancer in the Era of Immune Checkpoint Blockade.
Gastric cancer (GC) is one of the most important malignancies worldwide because of its high incidence and mortality. The very low survival rates are mainly related to late diagnosis and limited treatment options. GC is the final clinical outcome of a stepwise process that starts with a chronic and sustained inflammatory reaction mounted in response to Helicobacter pylori infection. The bacterium modulates innate and adaptive immunity presumably as part of the strategies to survive, which favors the creation of an immunosuppressive microenvironment that ultimately facilitates GC progression. T-cell exhaustion, which is characterized by elevated expression of immune checkpoint (IC) proteins, is one of the most salient manifestations of immunosuppressive microenvironments. It has been consistently demonstrated that the tumor-immune microenvironment(TIME)-exhausted phenotype can be reverted by blocking ICs with monoclonal antibodies. Although these therapies are associated with long-lasting response rates, only a subset of patients derive clinical benefit, which varies according to tumor site. The search for biomarkers to predict the response to IC inhibition is a matter of intense investigation as this may contribute to maximize disease control, reduce side effects, and minimize cost. The approval of pembrolizumab for its use in GC has rocketed immuno-oncology research in this cancer type. In this review, we summarize the current knowledge centered around the immune contexture and recent findings in connection with IC inhibition in GC.


Effects of speech on proofreading: can task-engagement manipulations shield against distraction?
This article reports 2 experiments that examine techniques to shield against the potentially disruptive effects of task-irrelevant background speech on proofreading. The participants searched for errors in texts that were either normal (i.e., written in Times New Roman font) or altered (i.e., presented either in Haettenschweiler font or in Times New Roman but masked by visual noise) in 2 sound conditions: a silent condition and a condition with background speech. Proofreading for semantic/contextual errors was impaired by speech, but only when the text was normal. This effect of speech was completely abolished when the text was written in an altered font (Experiment 1) or when it was masked by visual noise (Experiment 2). There was no functional difference between the 2 ways to alter the text with regard to the way the manipulations influenced the effects of background speech on proofreading. The results indicate that increased task demands, which lead to greater focal-task engagement, may shield against the distracting effects of background speech on proofreading.


M1 macrophages induce PD-L1hi cell-led collective invasion in HPV-positive head and neck squamous cell carcinoma via TNF-α/CDK4/UPS14.
Although the roles of PD-L1 in promoting tumor escape from immunosurveillance have been extensively addressed, its non-immune effects on tumor cells remain unclear. The spatial heterogeneity of PD-L1 staining in human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) tissues was identified by immunohistochemistry. Three-dimensional (3D) specific cell-led invasion assay and 3D cancer spheroid model were used to investigate the roles of PD-L1hileader cells in collective invasion. The impact of M1 macrophages on specific PD-L1 expression in leader cells and its mechanisms were further studied. Finally, the effect of combination therapy of anti-PD-L1 and CDK4 inhibitor on HPV-positive tumors were evaluated on a mice model. Here, we observed a distinctive marginal pattern of PD-L1 expression in HPV-positive HNSCC tissues. By mimicking this spatial pattern of PD-L1 expression in the 3D invasion assay, we found that PD-L1hi cells led the tumor collective invasion. M1 macrophages induced specific PD-L1 expression in leader cells, and depletion of macrophages in tumor-bearing mice abrogated PD-L1hileader cells and collective invasion. Mechanistically, TNF-α secreted by M1 macrophages markedly increased the abundance of PD-L1 via CDK4/ubiquitin-specific peptidase 14-mediated deubiquitination of PD-L1. We also found that suppression of CDK4 enhanced the efficacy of anti-PD-L1 therapy in an E6/E7 murine model. Our study identified TNF-α/CDK4/ubiquitin-specific peptidase 14-mediated PD-L1 stability as a novel mechanism underlying M1 macrophage-induced PD-L1hileader cells and collective tumor invasion, and highlighted the potential of the combination therapy of anti-PD-L1 and CDK4 inhibitor for HPV-positive HNSCC.


Compartmental fasciotomy and isolating a muscle from neighboring muscles interfere with myofascial force transmission within the rat anterior crural compartment.
Muscles within the anterior crural compartment (extensor digitorum longus, EDL; tibialis anterior, TA; and extensor hallucis longus, EHL) and within the peroneal compartment were excited simultaneously and maximally. All muscles were kept at constant length with the exception of EDL, for which muscle length was changed by moving its proximal tendon. Active and passive force was measured at proximal as well as distal EDL tendons and at the combined distal tendons of TA and EHL (TA+EHL). In the initial experimental condition, a difference (F(proximal) > F(distal)) in EDL force, amounting to 0-14% of proximal force, was confirmed for most EDL lengths. This is interpreted as a clear proof of extramuscular myofascial force transmission, as no significant EDL length effects could be shown on TA+EHL force. Repeated measurements were confirmed to cause marked changes of both proximal and distal length-force characteristics, such as a shift of the whole ascending limb of the active curve, including optimum length, to higher lengths without decreasing optimum force, and decreasing active force at low lengths (by approximately 57%). Repeated measurements also lowered proximal and distal EDL passive force (by up to 35%). The proximo-distal difference in passive as well as active EDL force was decreased, but persisted. At most lengths, this difference for active force amounted to a constant fraction (14%) of proximal force. TA+EHL force was not affected significantly. Subsequently, acute effects of experimental surgical alterations were studied: The first manipulation was full lateral fasciotomy of the anterior crural compartment that caused a further decrease in active force at the proximal EDL but not at the distal EDL tendon. Passive forces showed no further significant changes. The proximo-distal EDL active force difference decreased to 0-5% of proximal force. After fasciotomy, TA+EHL force increased by 30%. This was interpreted as evidence of increased intramuscular and decreased extramuscular myofascial force transmission. The second manipulation was full isolation of EDL from TA+EHL, but not from extramuscular connective tissues, which caused a further decrease of the EDL proximo-distal force differences, indicating a stiffening effect of the presence of TA+EHL on the extramuscular matrix. For EDL active force the difference was no longer significantly different from zero. In contrast, for EDL passive force the proximo-distal force difference persisted. It is concluded that extramuscular myofascial force transmission is an important feature of the anterior crural compartment. The magnitude of this force transmission requires that it be considered in analysis of muscular function.


The aging clock: circadian rhythms and later life.
Circadian rhythms play an influential role in nearly all aspects of physiology and behavior in the vast majority of species on Earth. The biological clockwork that regulates these rhythms is dynamic over the lifespan: rhythmic activities such as sleep/wake patterns change markedly as we age, and in many cases they become increasingly fragmented. Given that prolonged disruptions of normal rhythms are highly detrimental to health, deeper knowledge of how our biological clocks change with age may create valuable opportunities to improve health and longevity for an aging global population. In this Review, we synthesize key findings from the study of circadian rhythms in later life, identify patterns of change documented to date, and review potential physiological mechanisms that may underlie these changes.


Clinical inquiries. What is the best treatment for infants with colic?
Infantile colic, defined as excessive crying in an otherwise healthy baby, is a distressing phenomenon, but there is little evidence to support the many treatments offered. Several small studies report some benefit from use of a hypoallergenic (protein hydrolysate) formula, maternal diet adjustment (focusing on a low-allergen diet), and reduced stimulation of the infant. While dicyclomine has been shown to be effective for colic, there are significant concerns about its safety, and the manufacturer has contraindicated its use in this population. An herbal tea containing chamomile, vervain, licorice, fennel, and balm-mint was also effective in a small RCT, but the volume necessary for treatment limits its usefulness (strength of recommendation: B, inconsistent or limited-quality patient-oriented evidence). The one proven treatment is time, as this behavior tends to dissipate by 6 months of age.


Effects of inhaled corticosteroids on pneumonia severity and antimicrobial resistance.
Limited information is available regarding the impact of prior use of inhaled corticosteroids (ICS) in patients subsequently developing community-acquired pneumonia (CAP). We assessed the effects of prior ICS use on severity of illness and microbiology in CAP hospitalized patients. A retrospective cohort study of subjects with CAP (by the International Classification of Diseases, 9th Revision, Clinical Modification) was conducted over a 4-year period at 2 tertiary teaching hospitals. Subjects were considered to be ICS users if they received ICS prior to admission. Primary outcomes were severity of illness and microbiology at admission. Data were abstracted on 664 subjects: 89 prior ICS users (13.4%) and 575 non-users (86.6%). Prior ICS users had higher severity of illness at admission: mean ± SD Pneumonia Severity Index 100.8 ± 31.4 vs 68.8 ± 33.4, P = .001, and CURB-65 (confusion, urea nitrogen, respiratory rate, blood pressure, ≥ 65 years of age) score 1.56 ± 1.02 vs 1.19 ± 1.02, P = .002. Prior ICS use was independently associated with antimicrobial-resistant pathogens: 11.2% vs 5.9%, odds ratio 2.6, 95% CI 1.1-6.1, P = .04. Prior ICS use was associated with higher severity of illness at admission and higher incidence of antimicrobial-resistant pathogens in CAP hospitalized patients.


[Indirect calorimetry in mechanically ventilated children. 3. Clinical use of a new measurement procedure].
After the new device for measurement of oxygen uptake (VO2) and carbon dioxide elimination (VCO2) was tested in a pneumatic lung model, we investigated its clinical practicability. First the level of VO2 and VCO2 in the early postoperative phase was determined. Secondly we examined alterations in gasexchange caused by various intensive care activities. Whereby main concern was the change from mechanical ventilation to spontaneous CPAP-breathing. Ten children undergoing cardiac surgery (age: 10 days to 10 years) were included in the study. Three children were primarily investigated on the CPAP-system. Data from another six were collected under controlled ventilation as well as under spontaneous CPAP-breathing. One child had to be ventilated for a longer time (greater than 24 h). The change from mechanical ventilation to spontaneous breathing was accompanied with a temporary increase of VO2 and VCO2. As expected, all stressful factors like tracheal suction, pain or anxiety increased these two variables. Also some drugs showed remarkable changes. While marked increases under Normastigmine, Naloxone, and Dopamine could be observed, gas exchange was decreased under Morphine. However, heart rate showed no good correlation with VO2. We conclude that the developed prototype is a reliable device for measuring VO2 and VCO2. Our results suggest that spontaneous CPAP-breathing is not always accompanied with increased gas exchange. If there is an increased gas exchange, it may refer to transient changes during the adaption phase. Therefore, a continuous monitoring of VO2 and VCO2 over a longer period of time is necessary. Consequently the first phase after changing ventilatory support has to be viewed carefully. The heart rate in children, undergone cardiac surgery, is not a good predictor for determination of VO2 and thus for energy expenditure. This device allows to quantify accurately oxygen consumption and carbon dioxide elimination in ventilated children. Furthermore, alterations in metabolism and cardiopulmonary variables caused by intensive care and therapeutic interventions can be examined continuously. It seems to be a valuable tool for metabolic and cardiopulmonary monitoring in critically ill children.


Role of beta-lactam hydrolysis in the mechanism of resistance of a beta-lactamase-constitutive Enterobacter cloacae strain to expanded-spectrum beta-lactams.
Enterobacter cloacae strains producing chromosomally mediated beta-lactamase constitutively show high degrees of resistance to most of the third-generation beta-lactams. It has been proposed that this resistance is due to the nonhydrolytic binding or trapping of beta-lactams by the enzyme. We found that the outer membrane of E. cloacae strain 55M indeed had permeability to cefazolin about 14-fold lower than that of Escherichia coli, and that the number of beta-lactamase molecules produced by this constitutive mutant was exceptionally large (2 X 10(5) per cell). These conditions are expected to produce a low degree of resistance, but could not explain the high resistance level of the mutant. We showed that the beta-lactamase of this strain hydrolyzed third-generation beta-lactams at measurable rates. Although the V max for these compounds was less than 0.01% of that for cefazolin, the enzyme could hydrolyze them at rates comparable to the rate for cefazolin when the substrate concentration was near 0.1 microM, a concentration thought to be physiologically relevant for the inhibition of cell growth, because of the exceptionally high affinity of the enzyme to many third-generation compounds. Calculations based on kinetic parameters of the enzyme, outer membrane permeability, and affinity toward penicillin-binding proteins succeeded in predicting the MICs for several third-generation beta-lactams. The data suggest that hydrolysis may be more important than nonhydrolytic binding for the expression of the resistant phenotype, and that studies on the susceptibility of beta-lactams to beta-lactamases should be carried out at physiologically relevant, very low concentrations of the drug, rather than the customary very high concentrations, such as 100 microM.


Body Mass Index Is an Important Predictor for Suicide: Results from a Systematic Review and Meta-Analysis.
Public health concerns for the independent management of obesity and suicidal behavior are rising. Emerging evidence suggests body weight plays an important role in quantifying the risk of suicide. In light of these findings, we aimed to clarify the association between body mass index (BMI) and suicidal behavior by systematically reviewing and evaluating the literature. Studies were identified by searching MEDLINE, EMBASE, PsycINFO, and CINAHL from inception to January 2015, supplemented by hand and grey literature searches. Study screening, data extraction, and risk of bias assessment were conducted in duplicate. We included 38 observational studies. Meta-analyses supported an inverse association between BMI and completed suicide. Pooled summary estimates demonstrated that underweight was significantly associated with an increased risk of completed suicide (HR = 1.21, 95% CI 1.07 to 1.36, p = .002), and obesity (HR = 0.71, 95% CI 0.56 to 0.89, p = .003) and overweight (HR = 0.78, 95% CI 0.75 to 0.82, p < .0001) were significantly associated with a decreased risk of completed suicide relative to normal weight. A qualitative summary of the literature demonstrated conflicting evidence regarding the association between BMI and attempted suicide and revealed no association between BMI and suicidal ideation. BMI may be used to aid the assessment of suicide risk, especially that of completed suicide. However, unmeasured confounders and systematic biases of individual studies limit the quality of evidence.


Systems biology analysis of the proteomic alterations induced by MPP(+), a Parkinson's disease-related mitochondrial toxin.
Parkinson's disease (PD) is a complex neurodegenerative disease whose etiology has not been completely characterized. Many cellular processes have been proposed to play a role in the neuronal damage and loss: defects in the proteosomal activity, altered protein processing, increased reactive oxygen species burden. Among them, the involvement of a decreased activity and an altered disposal of mitochondria is becoming more and more evident. The mitochondrial toxin 1-methyl-4-phenylpyridinium (MPP(+)), an inhibitor of complex I, has been widely used to reproduce biochemical alterations linked to PD in vitro and its precursor, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP), to induce a Parkinson-like syndrome in vivo. Therefore, we performed a meta-analysis of the literature of all the proteomic investigations of neuronal alterations due to MPP(+) treatment and compared it with our results obtained with a mitochondrial proteomic analysis of SH-SY5Y cells treated with MPP(+). By using open-source bioinformatics tools, we identified the biochemical pathways and the molecular functions mostly affected by MPP(+), i.e., ATP production, the mitochondrial unfolded stress response, apoptosis, autophagy, and, most importantly, the synapse funcionality. Eventually, we generated protein networks, based on physical or functional interactions, to highlight the relationships among the molecular actors involved. In particular, we identified the mitochondrial protein HSP60 as the central hub in the protein-protein interaction network. As a whole, this analysis clarified the cellular responses to MPP(+), the specific mitochondrial proteome alterations induced and how this toxic model can recapitulate some pathogenetic events of PD.


Design and performance of a 24-station high throughput microbioreactor.
Two prototype 24-unit microbioreactors are presented and reviewed for their relative merits. The first used a standard 24-well plate as the template, while the second consisted of 24-discrete units. Both systems used non-invasive optical sensors to monitor pH and dissolved oxygen. The systems were used to cultivate Escherichia coli. Both designs had their merits and the results obtained are presented. In addition, dissolved oxygen control was demonstrated at the milliliter scale and 24 simultaneously monitored fermentations were successfully carried out. These results demonstrated high quality high throughput bioprocessing and provide important insights into operational parameters at small scale.


Migration of implanted markers for image-guided lung tumor stereotactic ablative radiotherapy.
The purpose of this study was to quantify postimplantation migration of percutaneously implanted cylindrical gold seeds ("seeds") and platinum endovascular embolization coils ("coils") for tumor tracking in pulmonary stereotactic ablative radiotherapy (SABR). We retrospectively analyzed the migration of markers in 32 consecutive patients with computed tomography scans postimplantation and at simulation. We implanted 147 markers (59 seeds, 88 coils) in or around 34 pulmonary tumors over 32 procedures, with one lesion implanted twice. Marker coordinates were rigidly aligned by minimizing fiducial registration error (FRE), the root mean square of the differences in marker locations for each tumor between scans. To also evaluate whether single markers were responsible for most migration, we aligned with and without the outlier causing the largest FRE increase per tumor. We applied the resultant transformation to all markers. We evaluated migration of individual markers and FRE of each group. Median scan interval was 8 days. Median individual marker migration was 1.28 mm (interquartile range [IQR] 0.78-2.63 mm). Median lesion FRE was 1.56 mm (IQR 0.92-2.95 mm). Outlier identification yielded 1.03 mm median migration (IQR 0.52-2.21 mm) and 1.97 mm median FRE (IQR 1.44-4.32 mm). Outliers caused a mean and median shift in the centroid of 1.22 and 0.80 mm (95th percentile 2.52 mm). Seeds and coils had no statistically significant difference. Univariate analysis suggested no correlation of migration with the number of markers, contact with the chest wall, or time elapsed. Marker migration between implantation and simulation is limited and unlikely to cause geometric miss during tracking.


Some film-noise measurements by diffraction of coherent light.
It is known that the diffraction of coherent light can be used for various spatial-noise measurements including the power-spectrum estimation of photographic emulsions. A number of such measurements are presented. The power spectrum of grain noise is compared with several theoretical models. The spectra of grain and phase noise for overexposed film are obtained. A simple procedure for obtaining the variance of the phase noise of photographic film, assuming gaussian statistics and statistical independence between grain and phase noise, is furnished.


Chromosomal deletion, gene amplification, alternative processing, and autocrine growth factor production in the pathogenesis of human lung cancer.
Molecular and cell biologic studies of a large number of lung cancer cell lines of all histologic types have revealed several mechanisms active in the pathogenesis of these cells. Small cell lung cancer (also called "oat cell" lung cancer) has a deletion involving chromosome region 3p(14-23) that is confirmed by DNA restriction fragment length polymorphisms analysis (studies done in collaboration with Dr. Susan Naylor). Several lung cancers of both small cell and non-small cell type (including adeno- and squamous cell lung cancer) express the proto-oncogenes c-, N-, or L-myc, and in some cases more than one of these family members. N-myc appears restricted in its expression to the small cell lung cancer type while c-myc and L-myc can be expressed in both small cell and non-small cell lung cancers. Many lung cancers of all histologic types also express large amounts of p53, which are not correlated with the amount or type of myc gene product expressed. In small cell lung cancer, high levels of myc gene expression are usually associated with gene amplification, and not uncommonly there is rearrangement of some of the amplified copies. In non-small cell lung cancer, expression without amplification or rearrangement of myc genes is seen. In contrast, high level expression of p53 is not associated with gene amplification in any lung cancer type. In addition, to these proto-oncogenes acting at a presumed nuclear locus, there is increased expression of various ras family members and the c-raf-1 proto-oncogene (in collaboration with Dr. Ulf Rapp). Lung cancer cells in tissue culture can grow in medium without serum and few or no other growth factors added. Thus, it appears that lung cancer cells can produce their own growth factors which can act in an "autocrine" fashion. The best characterized example of this is gastrin releasing peptide (GRP, also called bombesin) produced by small cell lung cancer. In at least some small cell lung cancers, interference with GRP action by specific monoclonal antibodies results in inhibition of tumor cell growth in culture and in nude mouse xenografts. Thus, constitutively expressed GRP gene may function as a cellular oncogene under certain circumstances in small cell lung cancer. Based on these observations we are proposing to test monoclonal anti-GRP antibodies in patients.


[A reflection on services to families in which the mother suffers from a mental disorder].
This article presents results of a research conducted in Québec City between 2004 and 2006. The objective of this study was to examine services to families in which the mother suffers from a mental disorder. The authors examine the experiences and perspectives of parents with a mental disorder, spouses and children on the services offered to families ; the study also examines the organisation, the basis and the context of actions and interactions between stakeholders of various sectors of services to families with a parent with severe mental illness. The authors conclude that despite Quebec's good intentions, members of families in which the mother suffers from a mental disorder do not benefit from a truly comprehensive family-focused approach.


Descriptive study of occupational therapists' participation in early childhood transitions.
The study describes the current role of occupational therapists in the transition of young children with special needs from early intervention to preschool and from preschool to kindergarten. Questionnaires were mailed to a random sample of 500 pediatric occupational therapists who are members of the American Occupational Therapy Association's School System Special Interest Section. The response rate was 44.3%, with 173 usable surveys. Approximately 40% of therapists working in the early-intervention-to-preschool transition and 32% of therapists working in the preschool-to-kindergarten transition reported not participating fully in the process. The primary barrier to full participation was "not enough time." Evaluation was identified as the most frequently used strategy for participation. Only 23% of respondents reported receiving specialized transition training. Although limited by a small sample, the study provides an initial description of occupational therapists' participation in early childhood transitions.


First insights into the biodiversity and biogeography of the Southern Ocean deep sea.
Shallow marine benthic communities around Antarctica show high levels of endemism, gigantism, slow growth, longevity and late maturity, as well as adaptive radiations that have generated considerable biodiversity in some taxa. The deeper parts of the Southern Ocean exhibit some unique environmental features, including a very deep continental shelf and a weakly stratified water column, and are the source for much of the deep water in the world ocean. These features suggest that deep-sea faunas around the Antarctic may be related both to adjacent shelf communities and to those in other oceans. Unlike shallow-water Antarctic benthic communities, however, little is known about life in this vast deep-sea region. Here, we report new data from recent sampling expeditions in the deep Weddell Sea and adjacent areas (748-6,348 m water depth) that reveal high levels of new biodiversity; for example, 674 isopods species, of which 585 were new to science. Bathymetric and biogeographic trends varied between taxa. In groups such as the isopods and polychaetes, slope assemblages included species that have invaded from the shelf. In other taxa, the shelf and slope assemblages were more distinct. Abyssal faunas tended to have stronger links to other oceans, particularly the Atlantic, but mainly in taxa with good dispersal capabilities, such as the Foraminifera. The isopods, ostracods and nematodes, which are poor dispersers, include many species currently known only from the Southern Ocean. Our findings challenge suggestions that deep-sea diversity is depressed in the Southern Ocean and provide a basis for exploring the evolutionary significance of the varied biogeographic patterns observed in this remote environment.


Congenital afibrinogenaemia caused by uniparental isodisomy of chromosome 4 containing a novel 15-kb deletion involving fibrinogen Aalpha-chain gene.
Among rare inherited deficiencies of coagulation factors, congenital afibrinogenaemia is characterised by the lack of fibrinogen in plasma. In the last few years, several genetic defects underlying afibrinogenaemia (mostly point mutations) have been described in the fibrinogen gene cluster. In this study, the molecular basis responsible for afibrinogenaemia in a Thai proband was defined. Point mutation screening was accomplished by directly sequencing the three fibrinogen genes. The impossibility to amplify fibrinogen Aalpha-chain gene (FGA) exons 5 and 6 suggested the presence of a homozygous deletion. A specific long-range PCR assay enabled the identification of a novel 15-kb deletion, representing the largest afibrinogenaemia-causing deletion described so far. Direct sequencing of the deletion junction allowed mapping of the breakpoints in FGA intron 4 and in the intergenic region between Aalpha- and Bbeta-chain genes. Since the mutation was inherited only from the mother and nonpaternity was ruled out, a maternal uniparental disomy (UPD) was hypothesised. UPD test, carried out with markers covering the whole chromosome 4, revealed that maternal isodisomy was responsible for homozygosity of the 15-kb deletion in the proband. The apparently normal phenotype of the proband, except for afibrinogenaemia, suggests that UPD for chromosome 4 is clinically silent. This represents the first case of a documented complete isodisomy of chromosome 4 causing the phenotypic expression of a recessive disorder. In silico analyses of the regions surrounding the breakpoints suggested that the 15-kb deletion might have originated from an inappropriate repair of a double-strand break by the nonhomologous end joining mechanism.


The impact of immigration detention on mental health: a systematic review.
The number of asylum seekers, refugees and internally displaced people worldwide has increased dramatically over the past 5 years. Many countries are continuing to resort to detaining asylum seekers and other migrants, despite concerns that this may be harmful. In light of the considerable body of recent research, this review aims to update and expand on a 2009 systematic review on the mental health consequences of detention on adult, adolescent and child immigration detainees, which found (on the basis on 9 studies) that there was consistent evidence that immigration detention had adverse effects on mental health. Three databases were searched using key terms relating to immigration detention and mental health. Electronic searches were supplemented by reference screening. Studies were included if they were quantitative, included individuals detained for immigration purposes, reported on mental health problems and were published in peer-reviewed journals. Two reviewers independently screened papers for eligibility, and a further two reviewers completed quality appraisals for included studies. Twenty- six studies (21 of which were not included in the 2009 review) reporting on a total of 2099 participants were included in the review. Overall, these studies indicated that adults, adolescents and children experienced high levels of mental health problems. Anxiety, depression and post-traumatic stress disorder were most commonly reported both during and following detention. Higher symptom scores were found in detained compared to non-detained refugees. In addition (and more clearly than was evident in 2009), detention duration was positively associated with severity of mental symptoms. Greater trauma exposure prior to detention was also associated with symptom severity. The literature base reviewed in this paper consistently demonstrated severe mental health consequences amongst detainees across a wide range of settings and jurisdictions. There is a pressing need for the proper consideration of mental health and consequent risk of detention-related harm in decisions surrounding detention as well as for improved care for individuals within detention facilities. Recommendations based on these findings are presented, including increased focus on the identification of vulnerability and on minimising the duration of detention.


Reduction of cytosolic p27(Kip1) inhibits cancer cell motility, survival, and tumorigenicity.
We generated a p27(Kip1) mutant (p27deltaNLS) that localized exclusively in cell cytosol. Expression of p27deltaNLS in MCF7 breast cancer cells down-regulated RhoA and increased motility, survival, and Akt levels without an effect on cell cycle distribution. RNA interference of p27 in U87 glioma cells, which express p27 predominantly in the cytoplasm, inhibited motility and survival. Conversely, knockdown of p27 in COS7 cells, with >95% nuclear p27 expression, accelerated proliferation but had no effect on motility or survival. U87 cells in which p27 had been eliminated by RNA interference exhibited lower Akt levels, shorter Akt turnover, and markedly impaired tumorigenicity in vivo. These xenografts were less invasive and exhibited increased apoptosis compared with p27-expressing tumors. Expression of cytosolic p27 in primary human breast carcinomas correlated linearly with Akt content as measured by immunohistochemistry. These data suggest that cytoplasmic p27 can exert oncogenic functions by modulating Akt stability, cell survival, and tumorigenicity.


Prospective study of dietary fiber, whole grain foods, and small intestinal cancer.
Although a number of epidemiologic studies have found dietary fiber and whole grains to be inversely associated with colorectal cancer incidence, studies of dietary and other risk factors for small intestinal cancer have been sparse and all of a case-control design. We conducted a prospective cohort study to determine the relationship between intake of dietary fiber/whole grains and the incidence of small intestinal cancer. We analyzed dietary data collected in 1995 and 1996 from 293,703 men and 198,618 women in the National Institutes of Health-AARP Diet and Health Study. We used multivariate Cox proportional hazards models to estimate relative risk (RR) and 2-sided 95% confidence intervals (CIs) for quintiles of dietary fiber and whole grain intake. Through 2003, 165 individuals developed small intestinal cancers. Dietary fiber/whole grain intake was generally associated with a lower risk of small intestinal cancer. The multivariate RRs (95% CIs; 5th vs 1st intake quintile) were 0.79 (0.43-1.44; P trend, .41) for total dietary fiber, 0.51 (0.29-0.89; P trend, .01) for fiber from grains, and 0.59 (0.33-1.05; P trend, .06) for whole grain foods. Intake of fiber from grains and whole-grain foods was inversely associated with small intestinal cancer incidence; the RR values were consistent with those from the same dietary factors for large bowel cancer in this cohort. In conjunction with the anatomic and physiologic commonalities of the large and small bowel, as well as the mutually increased risks for second cancer for both organs, grain fiber and whole grain foods seem to protect against lower gastrointestinal cancers.


Finger domain mutation affects enzyme activity, DNA replication efficiency, and fidelity of an exonuclease-deficient DNA polymerase of herpes simplex virus type 1.
The catalytic subunit of herpes simplex virus DNA polymerase (Pol), a member of the B family polymerases, possesses both polymerase and exonuclease activities. We previously demonstrated that a recombinant virus (YD12) containing a double mutation within conserved exonuclease motif III of the Pol was highly mutagenic and rapidly evolved to contain an additional leucine-to-phenylalanine mutation at residue 774 (L774F), which is located within the finger subdomain of the polymerase domain. We further demonstrated that the recombinant L774F virus replicated DNA with increased fidelity and that the L774F mutant Pol exhibited altered enzyme kinetics and impaired polymerase activity to extension from mismatched primer termini. In this study, we demonstrated that addition of the L774F mutation to the YD12 Pol did not restore the exonuclease deficiency. However, the polymerase activity of the YD12 Pol to extension from mismatched primer termini and on the nucleotide incorporation pattern was altered upon addition of the L774F mutation. The L774F mutation-containing YD12 Pol also supported the growth of viral progeny and replicated DNA more efficiently and more accurately than did the YD12 Pol. Together, these studies demonstrate that a herpes simplex virus Pol mutant with a highly mutagenic ability can rapidly acquire additional mutations, which may be selected for their survival and outgrowth. Furthermore, the studies demonstrate that the polymerase activity of HSV-1 Pol on primer extension is influenced by sequence context and that herpes simplex virus type 1 Pol may dissociate more frequently at G.C sites during the polymerization reaction. The implications of the findings are discussed.


American exceptionalism: population trends and flight initiation distances in birds from three continents.
All organisms may be affected by humans' increasing impact on Earth, but there are many potential drivers of population trends and the relative importance of each remains largely unknown. The causes of spatial patterns in population trends and their relationship with animal responses to human proximity are even less known. We investigated the relationship between population trends of 193 species of bird in North America, Australia and Europe and flight initiation distance (FID); the distance at which birds take flight when approached by a human. While there is an expected negative relationship between population trend and FID in Australia and Europe, we found the inverse relationship for North American birds; thus FID cannot be used as a universal predictor of vulnerability of birds. However, the analysis of the joint explanatory ability of multiple drivers (farmland breeding habitat, pole-most breeding latitude, migratory habit, FID) effects on population status replicated previously reported strong effects of farmland breeding habitat (an effect apparently driven mostly by European birds), as well as strong effects of FID, body size, migratory habit and continent. Farmland birds are generally declining. Flight initiation distance is related to population trends in a way that differs among continents opening new research possibilities concerning the causes of geographic differences in patterns of anti-predator behavior.


Minority recruitment into clinical trials: experimental findings and practical implications.
Racial and ethnic minorities in the US suffer disproportionately from obesity and related comorbidities, yet remain underrepresented in health research. To date, research on practical strategies to improve minority reach and recruitment into clinical trials is primarily descriptive rather than experimental. Within a randomized behavioral weight management trial for obese women, this recruitment experiment examined whether two characteristics of direct mail letters, an ethnically-targeted statement and personalization, increased the response rate among minority women. The ethnically-targeted statement noted ethnic-specific information about health risks of obesity. Personalized letters included recipients' names/addresses in the salutation and a handwritten signature on high-quality letterhead. Of women sent direct mail letters (N=30,000), those sent letters with the ethnically-targeted statement were more likely to respond than women sent letters with the generic statement, 0.8% (n=121) vs. 0.6% (n=90) respectively, p=.03, a 34.4% increase. Women sent personalized letters were no more likely to respond than women sent non-personalized letters, p=.53. In the weight management trial itself, of 267 women randomized into the trial, 33.7% (n=90) were minorities. Of minority women randomized into the trial, 68.9% (n=62) were recruited by direct mail letters: 75.8% (n=47) of those were sent a letter and 24.2% (n=15) were referred by friends/family who were sent a letter. The results indicate that a simple modification to a standard recruitment letter can have a meaningful impact on minority reach and recruitment rates. Practical implications include using ethnically-targeted, non-personalized direct mail letters and recruiting through friends/family at no additional cost.


Determination of basic strength of aliphatic amines through ion pair formation in some ionic liquid solutions.
To have an evaluation of the basic strength of aliphatic amines in ionic liquid solution, the stability constants relevant to the formation of amine/p-nitrophenol ion pairs were determined in different ionic liquids at 298 K. In particular, aliphatic (pyrrolidinium) and aromatic (imidazolium) ionic liquids were used. Imidazolium ions, bmim(+) and bm(2)im(+), having different hydrogen bond donor abilities were taken into account. Anions were chosen ([BF(4)(-)], [PF(6)(-)], and [NTf(2)(-)]; where NTf(2) = bis(trifluoromethansulfonyl)imide) showing different shape, size, and coordination ability. Several primary, secondary (cyclic or not), and tertiary amines were used to study the effect of amine structure on the ion pair stability. The comparison between data collected in this work and those previously obtained in ordinary molecular solvents evidence that ionic liquids are solvent media able to exalt the amine basicity. Furthermore, the experimental trends obtained in different ionic liquids can only be rationalized considering the whole of parameters affecting their tridimensional structure.


Direct investigation and accurate control of phase profile in liquid-crystal optical-phased array for beam steering.
An experimental setup and simple method were proposed to investigate and control the actual phase profile in a high-spatial-resolution liquid-crystal optical-phased array (LCOPA). A crossed polarizer and high-resolution microscope objective were employed to transform the light distribution out of the liquid-crystal layer into a polarization-interference pattern in which the phase-profile information was wrapped. The polarization-interference pattern was then directly translated into the actual phase profile. Based on this setup, a method was developed to accurately control the actual phase profile, and the steering efficiency at the steering angle of 16 mrad was increased from 80% to 90%. The proposed method also helps in increasing the steering efficiency when disclination lines appear.


The ultimate strength of glass silica nanowires.
In the past decade nanowires have attracted an increase interest because of their extraordinary mechanical strength. In fact, material properties in the nanoregime are extremely different from those found in macroscopic samples: few crystalline materials have shown a tensile strength in excess of 10 GPa in the form of nanowires. Still the length of defect-free crystalline nanowires is limited to a few millimeters and the strength of long nanowires is compromised by defects. The strength of glass nanowires is less affected by single defects. In this paper we present the ultimate strength of glass silica nanowires manufactured by a top-down fabrication technique; this is the highest value reported for glass materials. The measured ultimate strength is in excess of 10 GPa and increases for decreasing nanowire diameters. Scanning electron micrographs of the broken fragments showed a fragile rupture.


Successful Treatment for BK Virus Nephropathy by Leflunomide in a Kidney Transplant Patient: A Case Report.
The immunosuppressant agents in kidney transplantation (KT) may lead to various complications such as opportunistic infections and malignancies. BK virus associated nephropathy is a significant complication following KT, and it can result in graft failure. BK virus causes tubulointerstitial nephritis, ureter stenosis, and even graft failure in KT recipients with impaired immune system. We described a 63-year-old woman, who was a hepatitis C carrier and on dialysis for 22 years before KT, who received cadaveric-donor KT 2 years previously. She reported decreasing urine output and general weakness. The serum creatinine level was slightly increased from 2.94 to 4.38 mg/dL. Immunosuppressant medications including prednisolone, everolimus, cyclosporin, and mycophenolate sodium were continued as maintenance therapy post KT. Kidney biopsy was performed due to deterioration of graft function. The kidney biopsy showed consistent results with early-stage polyomavirus nephropathy, characterized by focal viral cytopathic changes with positive immunohistochemical signals and mesangial proliferative glomerulonephritis, immune-complex-mediated (Fig 1 and Fig 2). Negative C4d staining at peritubular capillary was reported. The dosage of mycophenolate sodium was tapered from 720 to 360 mg daily and that of everolimus increased from 0.5 to 1.0 mg daily due to BK viral infection with BK nephropathy. The serum creatinine level was 2.75 mg/dL after treatment. Early detection of BK nephropathy and decreasing immunosuppressant agents are the mainstay of treatment. Substituting leflunomide for mycophenolate sodium and increasing dosage of everolimus has been proposed to solve BK nephropathy. We presented that the use of leflunomide in such situation is in a timely manner.


Delusional depression. A distinct clinical entity?
The prognostic importance of delusions in the depressive syndrome had been a major focus of study before somatic therapies were available. Recently, the growing evidence that delusional depressives respond at a significantly lower rate to tricyclic antidepressants than do nondelusional depressives has revived this interest. That evidence is reviewed, and the demographic data and pretreatment clinical phenomenology of a series of hospitalized depressed patients were analyzed to see if differences existed between the delusional and nondelusional groups. Delusional unipolar depressives were less likely to recover while receiving placebo, had significantly more psychomotor retardation, and showed a trend toward fewer previous episodes than nondelusional unipolar depressives.


Intralesional triamcinolone acetonide injection for primary and recurrent chalazia: is it really effective?
To evaluate the safety and efficacy of intralesional triamcinolone acetonide (TA) injection in primary and recurrent chalazia. Retrospective, interventional, consecutive case series. One hundred forty-seven patients with primary or recurrent chalazia (155 cases) treated at the oculoplastic clinic at the Jules Stein Eye Institute between January 1, 2000, and December 31, 2003. Patients received an intralesional injection of 0.1 to 0.2 ml TA (40 mg/ml). Data regarding lesion size, including digital color photography, lesion regression or recurrence, and complete ophthalmic examination, were recorded at the time of injection and at different intervals until resolution or surgical excision. Success was defined as at least an 80% decrease in size with no recurrence. If the lesion recurred or regression was minimal (<50%), further injections were given as needed. Patients who declined injection or who did not respond to 2 to 3 injections were referred for surgical excision and drainage. Lesion size, clinical resolution, time to resolution, recurrence, and complications. Most of the patients received 1 injection (93 patients; 60%) or 2 injections (31 patients; 20%) with resolution of the lesion (more than 80% decrease in size), with an average time to resolution of 2.5 weeks. Patients who did not respond to 2 injections were more likely to fail treatment (minimal or no regression), to respond to further injections, or to undergo surgical excision and drainage (P = 0.0001, chi-square test). Patients with blepharitis required more injections to resolution (2+/-1.3 vs. 1.4+/-1; P = 0.05, independent samples t test). Intraocular pressure and visual acuity remained stable after treatment. No complications, such as visual loss, subcutaneous fat atrophy, or skin depigmentation changes, were noted with steroids injections; assuming a complication rate of 2%, our power was adequate to rule out these complications. Intralesional TA injection in primary and recurrent chalazia is effective in achieving lesion regression. Most cases resolve with an average of 1 to 2 injections. Chalazia that fail to respond to 2 or 3 injections are more likely to benefit from surgical excision. It may be considered as a first treatment in cases where diagnosis is straightforward.


Remodelling sympathetic innervation in rat pancreatic islets ontogeny.
Pancreatic islets are not fully developed at birth and it is not clear how they are vascularised and innervated. Nerve Growth Factor (NGF) is required to guide sympathetic neurons that innervate peripheral organs and also in cardiovascular system and ovary angiogenesis. Pancreatic beta cells of a transgenic mouse that over-expressed NGF in attracts sympathetic hyper-innervation towards them. Moreover, we have previously demonstrated that adult beta cells synthesize and secrete NGF; however, we do not know how is NGF secreted during development, nor if it might be trophic for sympathetic innervation and survival in the pancreas.We analyzed sympathetic innervation and vasculature development in rat pancreatic islets at different developmental stages; foetal (F19), early postnatal (P1), weaning period (P20) and adults. We temporarily correlated these events to NGF secretion by islet cells. Sympathetic fibres reached pancreatic islets in the early postnatal period, apparently following blood vessels. The maximal number of sympathetic fibres (TH immunopositive) in the periphery of the islets was observed at P20, and then fibres entered the islets and reached the core where beta cells are mainly located. The number of fibres decreased from that stage to adulthood. At all stages studied, islet cells secreted NGF and also expressed the high affinity receptor TrkA. Foetal and neonatal isolated islet cells secreted more NGF than adults. TrkA receptors were expressed at all stages in pancreatic sympathetic fibres and blood vessels. These last structures were NGF-immunoreactive only at early stages (foetal and P0). The results suggest that NGF signalling play an important role in the guidance of blood vessels and sympathetic fibres toward the islets during foetal and neonatal stages and could also preserve innervation at later stages of life.


Long-term Plans for Those Selecting Hospital Medicine as an Initial Career Choice.
The proportion of the newly graduated pediatric workforce that becomes hospitalists has been increasing slightly over the past decade. However, it is unknown what proportion of those who accept hospitalist positions as their first job intend to remain in the field longer term. This is important to workforce projections regarding the magnitude of those who will function in this role. The American Board of Pediatrics incorporated a structured questionnaire within the online application process to the General Pediatrics certification application. Respondents identified as residents or chief residents who selected "hospitalist position" as their immediate postresidency plan were the focus this study. We compared survey responses by gender and location of the medical school attended. Since the initiation of the general pediatrics certification examination application survey, 6335 completed the questionnaire. 79% (n=5001) were either in residency training or were a chief resident. Of those, 8% (n=376) reported they planned to work as a pediatric hospitalist immediately after completing residency. Fewer than half (43%; n=161) reported this to be their long-term career plan. This finding varied by both medical school type and by gender. The majority of pediatric residents and chief residents who take hospitalist positions immediately after training do not intend for hospital practice to be the long-term focus of their careers. As the field of hospital medicine continues to develop, understanding career trajectories can help inform current and future efforts regarding the potential for different mechanisms for training and certification.


Efficacy of intramammary tilmicosin and risk factors for cure of Staphylococcus aureus infection in the dry period.
The objective ofthis study was to evaluate the efficacy of intramammary tilmicosin, administered at drying-off, for eliminating Staphylococcus aureus infection, and to identify risk factors for S. aureus cure during the dry period. A total of 219 naturally infected cows, representing 308 quarters, were randomized to receive either one of two treatments at drying-off. Cows received either an intramammary infusion of 500 mg of benzathine cloxacillin, or a sterile solution containing 1,500 mg of tilmicosin. All cows had quarter milk samples taken aseptically three times before dry-off, and at wk 1, 2, and 4 of the subsequent lactation. Overall, 62% of cows and 67.5% of quarters infected with S. aureus cured during the dry period. The cure following administraton of tilmicosin was 67.3 and 72.5% for cows and quarters, respectively. By comparison, the cure achieved with cloxacillin was 56.9 and 62.9% of cows and quarters. Cows receiving tilmicosin were 2.1 times more likely to cure. The cure rate for cows decreased as the linear score on the last DHI test increased, and as the amount of S. aureus being shed increased. Quarters that cultured positive multiple times before drying-off were less likely to cure. Staphylococcus aureus infections located in front quarters of the udder were 2 times more likely to cure than those in hind quarters. Results of this study demonstrate that intramammary tilmicosin at drying-off is efficacious in curing existing S. aureus during the dry period. Risk factors associated with the cure of S. aureus were identified.


Medical misuse of controlled medications among adolescents.
To determine the past-year medical misuse prevalence for 4 controlled medication classes (pain, stimulant, sleeping, and antianxiety) among adolescents, and to assess substance use outcomes among adolescents who report medical misuse. A Web-based survey was self-administered by 2744 secondary school students in 2009-2010. Two southeastern Michigan school districts. The sample had a mean age of 14.8 years and was 51.1% female. The racial/ethnic distribution was 65.0% white, 29.5% African American, 3.7% Asian, 1.3% Hispanic, and 0.5% other. Past-year medical use and misuse of 4 controlled medication classes. Eighteen percent of the sample reported past-year medical use of at least 1 prescribed controlled medication. Among past-year medical users, 22.0% reported misuse of their controlled medications, including taking too much, intentionally getting high, or using to increase alcohol or other drug effects. Medical misusers were more likely than nonmisusers to divert their controlled medications and to abuse other substances. The odds of a positive screening result for drug abuse were substantially higher among medical misusers (adjusted odds ratio, 7.8; 95% confidence interval, 4.3-14.2) compared with medical users who used their controlled medications appropriately. The odds of drug abuse did not differ between medical users who used their controlled medications appropriately and nonusers. Most adolescents who used controlled medications took their medications appropriately. Substance use and diversion of controlled medications were more prevalent among adolescents who misused their controlled medications. Careful therapeutic monitoring could reduce medical misuse and diversion of controlled medications among adolescents.


X-ray diagnosable pneumonia in children with severe malnutrition at Kenyatta National Hospital.
To estimate the prevalence of radiologically evident pneumonia among children with severe malnutrition and to evaluate the diagnostic utility of commonly used clinical indicators of pneumonia among children with severe malnutrition. All children with severe malnutrition and admitted at the then Paediatric Observation Ward without congestive cardiac failure, severe anaemia, or severe dehydration, were clinically evaluated and a posteroanterior chest X-ray taken for each child. Pneumonia was diagnosed on the basis of radiological changes consistent with pneumonia as reported by an experienced radiologist. The performance of the various clinical parameters as diagnostic tests for pneumonia were also evaluated. Kenyatta National Hospital, a tertiary level teaching institution for the University of Nairobi. One hundred and seven children comprising 68 males and 39 females were recruited into the study. Of these children, 38 had kwashiorkor, 40 had marasmus, while 29 had marasmic kwashiorkor. Radiological evidence of pneumonia was found in 58% of children with kwashiorkor, 75% with marasmic kwashiorkor, and 82% with marasmus. All the commonly used clinical parameters performed poorly as diagnostic tests for pneumonia among children with severe malnutrition. Prevalence of pneumonia was very high among children with severe malnutrition. Available clinical parameters, singly or in combination, are poor diagnostic tools for pneumonia in children with severe malnutrition. It is advisable to treat children with severe malnutrition as if they had pneumonia, even in the absence of suggestive clinical signs.