Controlled self-assembly of alginate microgels by rapidly binding molecule pairs.
Controlled self-assembly of cell-encapsulating microscale polymeric hydrogels (microgels) could be advantageous in a variety of tissue engineering and regenerative medicine applications. Here, a method of assembly by chemical modification of alginate polymer with binding pair molecules (BPM) was explored. Alginate was modified with several types of BPM, specifically biotin and streptavidin and click chemistry compounds, and fabricated into 25-30 μm microgels using a microfluidic platform. These microgels were demonstrated to self-assemble under physiological conditions. By combining complementary microgels at a high ratio, size-defined assemblages were created, and the effects of BPM type and assembly method on the number of microgels per assemblage and packing density were determined. Furthermore, a magnetic process was developed to separate assemblages from single microgels, and allow formation of multilayer spheroids. Finally, cells were singly encapsulated into alginate microgels and assembled using BPM-modified alginate, suggesting potential applications in regenerative medicine.


Local public health system performance and community health outcomes.
Public health systems are charged with protecting the health and well-being of the communities they serve. Higher-performing public health systems should be associated with improved community health status. Currently, little research examines the impact of variations in public health system performance on community health outcomes. The current study seeks to provide local public health system partners with evidence regarding the impact of their efforts. It does so through examining the relationship between variations in local public health system performance in providing the core functions of public health (assessment, assurance, and policy development) and community health outcomes. Measures of community health status (dependent variables) and community demographic characteristics, collected from 1994 to 2003, were merged with measures of public health system performance collected from 2002 to 2007 (independent variables). For each dependent variable, Pearson correlations were calculated with all of the independent variables. Multivariate linear regression models were developed for each health outcome. Analysis was completed in 2010. Bivariate analysis found nine associations between local public health system performance and health outcomes. Linear regression models found associations between performance and three of the six outcomes measured. The outcomes associated with performance are those that would seem to be most sensitive to variations in practice. Poverty is significantly associated with all the health outcomes measured. These associations between performance and health outcomes suggest that improving performance of the core functions of public health may improve health.


Comparison of Cancer Yields and Diagnostic Performance of Screening Mammography vs. Supplemental Screening Ultrasound in 4394 Women with Average Risk for Breast Cancer.
The effectiveness of supplemental screening ultrasound (US) was investigated in women ≥ 40 years at average risk for breast cancer regardless of breast parenchymal density. A total of 4394 women at average risk and having previously undergone screening mammography were classified as the mammography group. Of 4394 women, 2005 underwent screening US after a final assessment of category 1 or 2 on screening mammography, and were categorized as the US group. Category 0, 4, and 5 on mammography and 3, 4, and 5 on US were defined as positive. The cancer yields per 1000 women and diagnostic performance of two groups were compared. The total cancer and invasive cancer yields for the mammography group were 3.0 (95 % confidence interval 1.6, 5.1) and 2.0 (95 % CI, 0.9, 3.9) per 1000 women, higher than the US values of 2.0 (0.5, 5.1) and 1.0 (0.1, 3.6), not statistically significant. The specificity, accuracy, and positive predictive value (PPV) for mammography were 88.90 % (87.93, 89.81), 88.85 % (87.88, 89.76), and 2.61 % (1.39, 4.41), significantly higher than the US values of 69.07 % (66.99, 71.09), 69.13 % (67.05, 71.15), and 0.64 % (0.18, 1.64). The short-term follow-up rate of mammography was 5.51 % (4.85, 6.22), significantly lower than the rate of 26.58 (24.66, 28.58) for US. Supplemental screening US in mammographically negative breasts can find additional carcinomas in women at average risk but is not as effective as screening mammography because of the lower cancer yield, invasive cancer yield, specificity, accuracy, PPV and a high short-term follow-up rate.


Human phenol sulfotransferases SULT1A2 and SULT1A1: genetic polymorphisms, allozyme properties, and human liver genotype-phenotype correlations.
Phenol sulfotransferases (PSTs or phenol SULTs) catalyze the sulfate conjugation of phenolic drugs, xenobiotics, and monoamines. Two human PST isoforms have been defined biochemically, a thermostable (TS), or phenol-preferring, and a thermolabile (TL), or monoamine-preferring form. Pharmacogenetic studies showed that levels of both TS PST activity and TS PST thermal stability (an indirect measure of variation in amino acid sequence) in the platelet were regulated by genetic polymorphisms. Subsequent molecular genetic experiments revealed the existence of three human PST genes, two of which, SULT1A1 and SULT1A2, encode proteins with "TS PST-like" activity. We recently reported common nucleotide polymorphisms for SULT1A1 that are associated with variations in platelet TS PST activity and thermal stability. In the present experiments, we set out to determine whether functionally significant DNA polymorphisms also might exist for SULT1A2, to compare the biochemical properties of all common allozymes encoded by SULT1A2 and SULT1A1, and to study phenol SULT genotype-phenotype correlations in the human liver. We phenotyped 61 human liver biopsy samples for TS PST thermal stability and activity. The open reading frames of SULT1A2 and SULT1A1 then were amplified with the polymerase chain reaction and sequenced for each of these hepatic tissue samples. We observed 13 SULT1A2 alleles that encoded 6 allozymes. These alleles were in linkage disequilibrium with alleles for SULT1A1. Biochemical characterization of common allozymes encoded by both genes suggested that SULT1A1 was primarily responsible for "TS PST phenotype" in the human liver. In summary, both SULT1A2 and SULT1A1 have a series of common alleles encoding enzymes that differ functionally and are associated with individual differences in phenol SULT properties in the liver.


High brain myo-inositol levels in the predementia phase of Alzheimer's disease in adults with Down's syndrome: a 1H MRS study.
An extra portion of chromosome 21 in Down's syndrome leads to a dementia in later life that is phenotypically similar to Alzheimer's disease. Down's syndrome therefore represents a model for studying preclinical stages of Alzheimer's disease. Markers that have been investigated in symptomatic Alzheimer's disease are myoinositol and N-acetyl-aspartate. The authors investigated whether abnormal brain levels of myo-inositol and other metabolites occur in the preclinical stages of Alzheimer's disease associated with Down's syndrome. The authors used 1H magnetic resonance spectroscopy (MRS) with external standards to measure absolute brain metabolite concentrations in 19 nondemented adults with Down's syndrome and 17 age- and sex-matched healthy comparison subjects. Concentrations of myoinositol and choline-containing compounds were significantly higher in the occipital and parietal regions of the adults with Down's syndrome than in the comparison subjects. Within the Down's syndrome group, older subjects (42-62 years, N = 11) had higher myo-inositol levels than younger subjects (28-39 years, N = 8). Older subjects in both groups had lower N-acetylaspartate levels than the respective younger subjects, although this old-young difference was not greater in the Down's syndrome group. The approximately 50% higher level of myo-inositol in Down's syndrome suggests a gene dose effect of the extra chromosome 21, where the human osmoregulatory sodium/myo-inositol cotransporter gene is located. The even higher myoinositol level in older adults with Down's syndrome extends to the predementia phase earlier findings of high myoinositol levels in symptomatic Alzheimer's disease.


Review: Structure, function and evolution of GnIH.
Neuropeptides that possess the Arg-Phe-NH2 motif at their C-termini (i.e., RFamide peptides) have been characterized in the nervous system of both invertebrates and vertebrates. In vertebrates, RFamide peptides make a family and consist of the groups of gonadotropin-inhibitory hormone (GnIH), neuropeptide FF (NPFF), prolactin-releasing peptide (PrRP), kisspeptin (kiss1 and kiss2), and pyroglutamylated RFamide peptide/26RFamide peptide (QRFP/26RFa). It now appears that these vertebrate RFamide peptides exert important neuroendocrine, behavioral, sensory, and autonomic functions. In 2000, GnIH was discovered as a novel hypothalamic RFamide peptide inhibiting gonadotropin release in quail. Subsequent studies have demonstrated that GnIH acts on the brain and pituitary to modulate reproductive physiology and behavior across vertebrates. To clarify the origin and evolution of GnIH, the existence of GnIH was investigated in agnathans, the most ancient lineage of vertebrates, and basal chordates, such as tunicates and cephalochordates (represented by amphioxus). This review first summarizes the structure and function of GnIH and other RFamide peptides, in particular NPFF having a similar C-terminal structure of GnIH, in vertebrates. Then, this review describes the evolutionary origin of GnIH based on the studies in agnathans and basal chordates.


Functional properties of ovotransferrin from chicken egg white and its derived peptides: a review.
With emerging trends in the food and pharmaceutical industries, potential applications of egg-derived bioactive compounds were recognized. Ovotransferrin is a major egg white functional protein responsible for multiple bioactivities. The objectives of this review are to provide scientific evidence of the functional properties of chicken ovotransferrin and its derived peptides and to identify future research approaches and applications. Various easy, economical, and non-toxic methods have been reported to produce ovotransferrin with high yield and purity, and chemical and enzymatic approaches have been employed to release bioactive peptides. The native ovotransferrin is known to have antimicrobial, antioxidant, anticancer, and immunomodulatory activities. The peptides produced from ovotransferrin also are reported to have antioxidant, antimicrobial, antihypertensive, and anticancer properties. However, little or no application of these compounds in the food and pharmaceutical areas is available yet. Therefore, the practical application of OTF in nutraceutical and pharmaceutical areas are among the emerging areas of research.


Budget Impact Analysis of Dalbavancin in the Treatment of Acute Bacterial Skin and Skin Structure Infections in Three European Countries.
Acute bacterial skin and skin structure infections (ABSSSIs) have been defined by the US Food and Drug Administration (FDA) in 2013 to include a subset of complicated skin and skin structure infections commonly treated with parenteral antibiotic therapy. Inpatient treatment of ABSSSIs involves a significant economic burden on the healthcare system. This study aimed to evaluate the economic impact on the National Health System associated with the management of non-severe ABSSSIs treated in hospitals with innovative long-acting dalbavancin compared to standard antibiotic therapy in Italy, Spain, and Austria. A budget impact analysis was developed to evaluate the direct costs associated with the management of ABSSSI from the national public health system perspective. The model considered the possibility of early discharge of patients directly from the Emergency Department (ED), after 1 night in the hospital, or after two or three nights in the hospital. A scenario with Standard of Care was compared with a dalbavancin scenario, where patients had the possibility of being discharged early. The epidemiological and cost parameters were extrapolated from national administrative databases and from a systematic literature review for each country. The analysis was conducted in a 3-year time horizon. A one-way deterministic sensitivity analysis was conducted to examine the robustness of the results. The model estimated an average annual number of patients with non-severe ABSSSI in Italy, Spain, and Austria equal to 5396, 7884, and 1788, respectively. A total annual expenditure of about €9.9 million, €13.5 million, and €3.4 million was estimated for treating the full set of ABSSSI patients in Italy, Spain, and Austria, respectively. Dalbavancin reduced the in-hospital length of stay in each country. In the first year of its introduction, dalbavancin significantly reduced the total economic burden in Italy and Spain (- €352,252 and - €233,991, respectively), while it increased the total economic burden in Austria (€80,769, 0.7% of the total expenditure for these patients); in the third year of its introduction, dalbavancin reduced the total economic burden in each Country (- €1.1 million, - €810,650, and - €70,269, respectively). The introduction of dalbavancin in a new patient pathway to treat non-severe ABSSSI could generate a significant reduction in hospitalized patients and the overall patient length of stay in hospital.


Comparative Genomics of Escherichia coli Sequence Type 219 Clones From the Same Patient: Evolution of the IncI1 blaCMY-Carrying Plasmid in Vivo.
This study investigates the evolution of an Escherichia coli sequence type 219 clone in a patient with recurrent urinary tract infection, comparing isolate EC974 obtained prior to antibiotic treatment and isolate EC1515 recovered after exposure to several β-lactam antibiotics (ceftriaxone, cefixime, and imipenem). EC974 had a smooth colony morphology, while EC1515 had a rough colony morphology on sheep blood agar. RAPD-PCR analysis suggested that both isolates belonged to the same clone. Antimicrobial susceptibility tests showed that EC1515 was more resistant to piperacillin/tazobactam, cefepime, cefpirome, and ertapenem than EC974. Comparative genomic analysis was used to investigate the genetic changes of EC974 and EC1515 within the host, and showed three plasmids with replicons IncI1, P0111, and IncFII in both isolates. P0111-type plasmids pEC974-2 and pEC1515-2, contained the antibiotic resistance genes aadA2, tetA, and drfA12. IncFII-type plasmids pEC974-3 and pEC1515-3 contained the antibiotic resistance genes blaTEM-1, aadA1, aadA22, sul3, and inuF. Interestingly, blaCMY-111 and blaCMY-4 were found in very similar IncI1 plasmids that also contained aadA22 and aac(3)-IId, from isolates EC974 (pEC974-1) and EC1515 (pEC1515-1), respectively. The results showed in vivo amino acid substitutions converting blaCMY-111 to blaCMY-4 (R221W and A238V substitutions). Conjugation experiments showed a high frequency of IncI1 and IncFII plasmid co-transference. Transconjugants and DH5α cells harboring blaCMY-4 or blaCMY-111 showed higher levels of resistance to ampicillin, amoxicillin, cefazolin, cefuroxime, cefotaxime, cefixime, and ceftazidime, but not piperacillin/tazobactam, cefpime, or ertapenem. All known genes (outer membrane proteins and extended-spectrum AmpC β-lactamases) involved in ETP resistance in E. coli were identical between EC974 and EC1515. This is the first study to identify the evolution of an IncI1 plasmid within the host, and to characterize blaCMY-111 in E. coli.


The central nucleus of the amygdala contributes to the production of morphine antinociception in the rat tail-flick test.
Current models of endogenous pain control circuitry emphasize neural substrates within the brainstem and spinal cord. We have recently shown, however, that the central nucleus of the amygdala (Ce) contributes to morphine-induced suppression of formalin-induced nociceptive behaviors. In the four experiments reported here, we investigated the possibility that the Ce also contributes to morphine-induced suppression of simple, spinally mediated nociceptive reflexes. Bilateral N-methyl-D-aspartate (NMDA)-induced lesions of the rat Ce, but not bilateral lesions centered on either the basolateral or medial amygdaloid nucleus, abolished the antinociception produced by 2.5 mg/kg morphine sulfate in the noxious heat-evoked tail-flick test. Bilateral Ce lesions also abolished the antinociception produced by 2 or 4 mg/kg morphine sulfate, but a relatively large dose of morphine sulfate (10 mg/kg, s.c.) resulted in partial reinstatement of antinociception. It is unlikely that these effects were due to secondary, seizure-induced damage following NMDA injection (e.g., to areas outside the amygdala) since bilateral inactivation of the Ce with the local anesthetic lidocaine also reliably attenuated morphine antinociception. It is also unlikely that these effects were artifacts of lesion-induced hyperalgesia, since Ce lesions failed to result in reliable thermal hyperalgesia, even at baseline tail-flick latencies of 10-12 sec. These data are the first to provide direct evidence that systemically administered morphine requires the integrity of a forebrain area in order to suppress spinally mediated nociceptive reflexes. It is argued that the present results, together with recent evidence linking the Ce to the production of several forms of conditioned and unconditioned environmentally induced antinociception, warrant incorporation of the Ce into current models of endogenous pain control circuitry.


Evidence of widespread binding of HLA class I molecules to peptides.
We have tested the binding of HLA class I proteins to peptides using a solid-phase binding assay. We tested 102 peptides, mostly derived from the HIV gag and HIV pol sequences. Most peptides did not bind to any class I protein tested. The pattern of binding among the three class I proteins tested, HLA-A2, -B27, and -B8, was approximately 85% concordant. Further, all five of the known HIV-1 gag T cell epitopes detected by human CTL bound at least one class I protein. Binding of class I to the peptides could be detected either by directly iodinated class I proteins, or indirectly using monoclonal antibodies specific for class I. The binding to the plates could be blocked with MA2.1, which binds in the alpha 1 region of A2, but not by W6/32, which binds elsewhere. The data presented here show that binding of class I to peptides is specific, but that many peptides bind to more than a single class I protein.


Duplex scanning of the portal vein and portasystemic shunts.
This article describes the preliminary findings of a Duplex ultrasound technique that enables the patency of the portal circulation and portosystemic shunt patency to be determined. In addition, the equipment allows the noninvasive measurement of quantitative rates of blood flow in these vessels. Portal vein flow was 952 +/- 273 ml/min in 10 normal subjects after an overnight fast and increased by 50% at 30 minutes in response to a standard 660-calorie liquid meal. Thirteen portosystemic shunts were scanned, and hemodynamic information was obtained from 10. Three of the four patients who had a Warren shunt performed 3 to 4 years earlier had portal vein occlusion. There is evidence of an increase in flow through the Warren shunt on feeding, suggesting a hemodynamic connection to the mesenteric side of the circulation. The apparatus, technique of examination, and the characteristics and difficulties with particular types of shunts are described.


How vocabulary size in two languages relates to efficiency in spoken word recognition by young Spanish-English bilinguals.
Research using online comprehension measures with monolingual children shows that speed and accuracy of spoken word recognition are correlated with lexical development. Here we examined speech processing efficiency in relation to vocabulary development in bilingual children learning both Spanish and English (n=26 ; 2 ; 6). Between-language associations were weak: vocabulary size in Spanish was uncorrelated with vocabulary in English, and children's facility in online comprehension in Spanish was unrelated to their facility in English. Instead, efficiency of online processing in one language was significantly related to vocabulary size in that language, after controlling for processing speed and vocabulary size in the other language. These links between efficiency of lexical access and vocabulary knowledge in bilinguals parallel those previously reported for Spanish and English monolinguals, suggesting that children's ability to abstract information from the input in building a working lexicon relates fundamentally to mechanisms underlying the construction of language.


Hypoxia index in the prediction of abnormal CTG at delivery in uncomplicated pregnancies.
The evaluation of hypoxia index (HI) in the prediction of abnormal fetal heart rate at delivery in uncomplicated pregnancies. The study group included 148 pregnant patients at term (69 patients with and 79 without brain sparing effect). The value of C/U ratio and HI was determined. Next, its value in predicting abnormal fetal heart rate during labor was evaluated. Then the predictive value of the HI index, C/U and last abnormal values of PI and RI in the MCA and the UA were compared in relation to the analyzed parameters. Evaluation included signs of fetal distress in CTG and abnormal fetal outcome. Then selected parameters, characterizing pregnancy course and fetal outcome with abnormal Doppler indices, were compared. The designated value of hypoxia index characterized by abnormal neonatal outcome was >10 for sHI and >14 for HI. Low prognostic value of MCA PI and RI, and UA PI and RI has been shown. The highest predictive value was marked by C/U and HI. There were no statistically significant differences in prediction of abnormal fetal heart rate during labor between C/U ratio, HI and sHI. The C/U ratio showed the highest sensitivity in the prediction of fetal abnormal heart rate. The C/U ratio, as a easier test, should be recommended as a first-line test in the prediction of abnormal CTG recordings in uncomplicated pregnancies.


Functional morphology and bite performance of raptorial chelicerae of camel spiders (Solifugae).
Solifugae are an understudied group of relatively large arachnids with well over 1000 species distributed on almost all major continents. These highly active predators utilize their large chelicerae for feeding, defense, burrowing and mating. We investigated the differences in cheliceral morphology and performance of two ecologically divergent species from North Africa; the cursorial Galeodes sp. and the burrowing Rhagodes melanus. Morphological data show differences in aspect ratio between the two species. Bite force measurements show Rhagodes (N=11) to be a much stronger biter than Galeodes (N=8), in terms of both absolute maximum force (Rhagodes 5.63 N, Galeodes 2.12 N) and force relative to cheliceral size. Synchrotron microtomographs of one specimen for each species reveal large differences in physiological cross-sectional area (PCSA) and estimated muscle stress, resulting in a much higher muscle stress in Rhagodes. This species also showed a longer muscle fiber length. Muscle volume and PCSA were found to differ between the two chelicerae in the two scanned specimens. Whereas Rhagodes reflects this morphological asymmetry in having a higher bite force in the right chelicera, Galeodes shows no such bias.


Central nervous system stem cell transplantation for children with neuronal ceroid lipofuscinosis.
Infantile and late-infantile neuronal ceroid lipofuscinoses (NCLs) are invariably fatal lysosomal storage diseases associated with defects in lysosomal enzyme palmitoyl-protein thioesterase 1 (PPT-1) or tripeptidyl peptidase 1 (TPP1) activity. Previous preclinical studies have demonstrated that human CNS stem cells (HuCNS-SCs) produce both PPT-1 and TPP1 and result in donor cell engraftment and reduced accumulation of storage material in the brain when tested in an NCL mouse model. HuCNS-SC transplantation was tested in an open-label dose-escalation Phase I clinical trial as a potential treatment for infantile and late-infantile NCL. Study design included direct neurosurgical transplantation of allogeneic HuCNS-SCs into the cerebral hemispheres and lateral ventricles accompanied by 12 months of immunosuppression. Six children with either the infantile or late-infantile forms of NCL underwent low- (3 patients) and high- (3 patients) dose transplantation of HuCNS-SCs followed by immunosuppression. The surgery, immunosuppression, and cell transplantation were well tolerated. Adverse events following transplantation were consistent with the underlying disease, and none were directly attributed to the donor cells. Observations regarding efficacy of the intervention were limited by the enrollment criteria requiring that patients be in advanced stages of disease. This study represents the first-in-human clinical trial involving transplantation of a purified population of human neural stem cells for a neurodegenerative disorder. The feasibility of this approach and absence of transplantation-related serious adverse events support further exploration of HuCNS-SC transplantation as a potential treatment for select subtypes of NCL, and possibly for other neurodegenerative disorders.


Effects of polyamines on Vibrio cholerae virulence properties.
Vibrio cholerae is the causative agent of the severe enteric disease cholera. To cause cholera the bacterium must be able to synthesize both cholera toxin (CT) and toxin-coregulated pilus (TCP) which mediates autoagglutination and is required for colonization of the small intestine. Only a few environmental signals have been shown to regulate V. cholerae virulence gene expression. Polyamines, which are ubiquitous in nature, and have been implicated in regulating virulence gene expression in other bacteria, have not been extensively studied for their effect on V. cholerae virulence properties. The objective of this study was to test the effect of several polyamines that are abundant in the human intestine on V. cholerae virulence properties. All of the polyamines tested inhibited autoagglutination of V. cholerae O1 classical strain in a concentration dependent manner. Putrescine and cadaverine decreased the synthesis of the major pilin subunit, TcpA, spermidine increased its production, and spermine had no effect. Putrescine and spermidine led to a decrease and increase, respectively, on the relative abundance of TCP found on the cell surface. Spermine led to a small reduction in cholera toxin synthesis whereas none of the other polyamines had an effect. The polyamines did not affect pili bundling morphology, but caused a small reduction in CTXφ transduction, indicating that the TCP present on the cell surface may not be fully functional. We hypothesize the inhibition of autoagglutination is likely to be caused by the positively charged amine groups on the polyamines electrostatically disrupting the pili-pili interactions which mediate autoagglutination. Our results implicate that polyamines may have a protective function against colonization of the small intestine by V. cholerae.


Increases of intracellular magnesium promote glycodeoxycholate-induced apoptosis in rat hepatocytes.
Retention of bile salts by the hepatocyte contributes to liver injury during cholestasis. Although cell injury can occur by one of two mechanisms, necrosis versus apoptosis, information is lacking regarding apoptosis as a mechanism of cell death by bile salts. Our aim was to determine if the bile salt glycodeoxycholate (GDC) induces apoptosis in rat hepatocytes. Morphologic assessment included electron microscopy and quantitation of nuclear fragmentation by fluorescent microscopy. Biochemical studies included measurements of DNA fragmentation, in vitro endonuclease activity, cytosolic free Ca2+ (Cai2+), and cytosolic free Mg2+ (Mgi2+). Morphologic studies demonstrated typical features of apoptosis in GDC (50 microM) treated cells. The "ladder pattern" of DNA fragmentation was also present in DNA obtained from GDC-treated cells. In vitro endonuclease activity was 2.5-fold greater with Mg2+ than Ca2+. Although basal Cai2+ values did not change after addition of GDC, Mgi2+ increased twofold. Incubation of cells in an Mg(2+)-free medium prevented the rise in Mgi2+ and reduced nuclear and DNA fragmentation. In conclusion, GDC induces apoptosis in hepatocytes by a mechanism promoted by increases of Mgi2+ with stimulation of Mg(2+)-dependent endonucleases. These data suggest for the first time that changes of Mgi2+ may participate in the program of cellular events culminating in apoptosis.


Development. A degrading way to make an organ.
Working out how organs form during embryonic development is a fascinating area of research. In a witty Perspective, Jeff Hardin describes new findings (Nishiwaki et al.) that reveal the many intricate steps needed for gonads to form in the worm C. elegans. Two key players, GON-1 and MIG-17, are metalloproteases that may help migration of distal tip cells by degrading extracellular matrix components.


Photographic gradient singlets.
Previous investigations involving the design of gradient lenses have been based mostly on third-order aberrations. The present paper discusses design techniques based on total aberrations. In particular, an algebraic formula is found for the total meridional curvature of radial gradient singlets. The formula obtained holds for a large class of refractive-index profiles.


Marginal and Internal Adaptation of Zirconia Crowns: A Comparative Study of Assessment Methods.
Marginal and internal adaptation is critical for the success of indirect restorations. New imaging systems make it possible to evaluate these parameters with precision and non-destructively. This study evaluated the marginal and internal adaptation of zirconia copings fabricated with two different systems using both silicone replica and microcomputed tomography (micro-CT) assessment methods. A metal master model, representing a preparation for an all-ceramic full crown, was digitally scanned and polycrystalline zirconia copings were fabricated with either Ceramill Zi (Amann-Girrbach) or inCoris Zi (Dentslpy-Sirona), n=10. For each coping, marginal and internal gaps were evaluated by silicone replica and micro-CT assessment methods. Four assessment points of each replica cross-section and micro-CT image were evaluated using imaging software: marginal gap (MG), axial wall (AW), axio-occlusal angle (AO) and mid-occlusal wall (MO). Data were statistically analyzed by factorial ANOVA and Tukey test (a=0.05). There was no statistically significant difference between the methods for MG and AW. For AO, there were significant differences between methods for Amann copings, while for Dentsply-Sirona copings similar values were observed. For MO, both methods presented statistically significant differences. A positive correlation was observed determined by the two assessment methods for MG values. In conclusion, the assessment method influenced the evaluation of marginal and internal adaptation of zirconia copings. Micro-CT showed lower marginal and internal gap values when compared to the silicone replica technique, although the difference was not always statistically significant. Marginal gap and axial wall assessment points showed the lower gap values, regardless of ceramic system and assessment method used.


Human Glycerol 3-Phosphate Dehydrogenase: X-ray Crystal Structures That Guide the Interpretation of Mutagenesis Studies.
Human liver glycerol 3-phosphate dehydrogenase ( hlGPDH) catalyzes the reduction of dihydroxyacetone phosphate (DHAP) to form glycerol 3-phosphate, using the binding energy associated with the nonreacting phosphodianion of the substrate to properly orient the enzyme-substrate complex within the active site. Herein, we report the crystal structures for unliganded, binary E·NAD, and ternary E·NAD·DHAP complexes of wild type hlGPDH, illustrating a new position of DHAP, and probe the kinetics of multiple mutant enzymes with natural and truncated substrates. Mutation of Lys120, which is positioned to donate a proton to the carbonyl of DHAP, results in similar increases in the activation barrier to hlGPDH-catlyzed reduction of DHAP and to phosphite dianion-activated reduction of glycolaldehyde, illustrating that these transition states show similar interactions with the cationic K120 side chain. The K120A mutation results in a 5.3 kcal/mol transition state destabilization, and 3.0 kcal/mol of the lost transition state stabilization is rescued by 1.0 M ethylammonium cation. The 6.5 kcal/mol increase in the activation barrier observed for the D260G mutant hlGPDH-catalyzed reaction represents a 3.5 kcal/mol weakening of transition state stabilization by the K120A side chain and a 3.0 kcal/mol weakening of the interactions with other residues. The interactions, at the enzyme active site, between the K120 side chain and the Q295 and R269 side chains were likewise examined by double-mutant analyses. These results provide strong evidence that the enzyme rate acceleration is due mainly or exclusively to transition state stabilization by electrostatic interactions with polar amino acid side chains.


Rapid and Efficient Isolation of Exosomes by Clustering and Scattering.
Tumor-derived extracellular vesicles (EVs) have become important biomarkers of liquid biopsies for precision medicine. However, the clinical application of EVs has been limited due to the lack of EV isolation practical technology applicable to clinical environments. Here, we report an innovative EV isolation method, which is quick and simple, and facilitates high-yield and high-purity EV isolation from blood. Introducing a cationic polymer in plasma resulted in rapid clustering of anionic EVs and a chaotropic agent can separate EVs from these clusters. Isolated EVs were characterized in terms of size distribution, morphology, surface protein markers, and exosomal RNA. Through performance comparison with various methods, including ultracentrifugation (UC), the present method delivered the highest recovery rate (~20 folds that of UC) and purity ratio (3.5 folds that of UC) of EVs in a short period of time (<20 min). The proposed method is expected to be used in basic and applied research on EV isolation and in clinical applications.


Twiddler's syndrome in a patient with a deep brain stimulation device for generalized dystonia.
Deep brain stimulation (DBS) is the technique of neurostimulation of deep brain structures for the treatment of conditions such as essential tremor, dystonia, Parkinson's disease and chronic pain syndromes. The procedure uses implanted deep brain stimulation electrodes connected to extension leads and an implantable pulse generator (IPG). Hardware failure related to the DBS procedure is not infrequent, and includes electrode migration and disconnection. We describe a patient who received bilateral globus pallidus internus DBS for dystonia with initially good clinical response, but the device eventually failed. Radiographs showed multiple twisting of the extension leads with disconnection from the brain electrodes and a diagnosis of Twiddler's syndrome was made. Twiddler's syndrome was first described in patients with cardiac pacemakers. Patients with mental disability, elderly and obese patients are at increased risk. Twiddler's syndrome should be suspected whenever there is a failure of the DBS device to relieve symptoms previously responsive to stimulation. Surgical correction is usually required.


Selective Androgen Receptor Modulator RAD140 Inhibits the Growth of Androgen/Estrogen Receptor-Positive Breast Cancer Models with a Distinct Mechanism of Action.
Purpose: Steroidal androgens suppress androgen receptor and estrogen receptor positive (AR/ER+) breast cancer cells and were used to treat breast cancer, eliciting favorable response. The current study evaluates the activity and efficacy of the oral selective AR modulator RAD140 in in vivo and in vitro models of AR/ER+ breast cancer.Experimental Design: A series of in vitro assays were used to determine the affinity of RAD140 to 4 nuclear receptors and evaluate its tissue-selective AR activity. The efficacy and pharmacodynamics of RAD140 as monotherapy or in combination with palbociclib were evaluated in AR/ER+ breast cancer xenograft models.Results: RAD140 bound AR with high affinity and specificity and activated AR in breast cancer but not prostate cancer cells. Oral administration of RAD140 substantially inhibited the growth of AR/ER+ breast cancer patient-derived xenografts (PDX). Activation of AR and suppression of ER pathway, including the ESR1 gene, were seen with RAD140 treatment. Coadministration of RAD140 and palbociclib showed improved efficacy in the AR/ER+ PDX models. In line with efficacy, a subset of AR-repressed genes associated with DNA replication was suppressed with RAD140 treatment, an effect apparently enhanced by concurrent administration of palbociclib.Conclusions: RAD140 is a potent AR agonist in breast cancer cells with a distinct mechanism of action, including the AR-mediated repression of ESR1 It inhibits the growth of multiple AR/ER+ breast cancer PDX models as a single agent, and in combination with palbociclib. The preclinical data presented here support further clinical investigation of RAD140 in AR/ER+ breast cancer patients. Clin Cancer Res; 23(24); 7608-20. ©2017 AACR.


Zero or not? Causes and consequences of zero-flow stream gage readings.
Streamflow observations can be used to understand, predict, and contextualize hydrologic, ecological, and biogeochemical processes and conditions in streams. Stream gages are point measurements along rivers where streamflow is measured, and are often used to infer upstream watershed-scale processes. When stream gages read zero, this may indicate that the stream has fully dried; however, zero-flow readings can also be caused by a wide range of other factors. Our ability to identify whether or not a zero-flow gage reading indicates a dry fluvial system has far reaching environmental implications. Incorrect identification and interpretation by the data user can lead to hydrologic, ecological, and/or biogeochemical predictions from models and analyses. Here, we describe several causes of zero-flow gage readings: frozen surface water, flow reversals, instrument error, and natural or human-driven upstream source losses or bypass flow. For these examples, we discuss the implications of zero-flow interpretations. We also highlight additional methodss for determining flow presence, including direct observations, statistical methods, and hydrologic models, which can be applied to interpret causes of zero-flow gage readings and implications for reach- and watershed-scale dynamics. Such efforts are necessary to improve our ability to understand and predict surface flow activation, cessation, and connectivity across river networks. Developing this integrated understanding of the wide range of possible meanings of zero-flows will only attain greater importance in a more variable and changing hydrologic climate.


Prospective evaluation of the self-pressurized air-Q intubating laryngeal airway in children.
To assess the clinical efficacy of the self-pressurized air-Q ILA™ (ILA-SP). The purpose of this prospective audit was to evaluate the feasibility of the ILA-SP in clinical practice and generate data for future comparison trials. The ILA-SP is a new first-generation supraglottic airway for children with a self-adjusting cuff and lack of a pilot balloon. Over a 4-month period, 352 children with an ASA physical status of I-III, newborn to 18 years of age, undergoing various procedures were studied. Data points assessed included insertion success rates, airway leak pressures, quality of ventilation, and perioperative complications associated with the use of this device. In 349 of the 352 patients in this study, the ILA-SP was used successfully as a primary supraglottic airway device in a variety of patients. Three patients required conversion to a standard laryngeal mask airway or a tracheal tube. The mean initial airway leak pressure for all patients was 17.8 ± 5.4 cm H(2)O, and 20.4 ± 5.5 cm H(2)O when re-checked at 10 min, which was statistically significant (P < 0.001). Complications were limited to 14 patients and related to reflex activation of the airway (coughing, laryngospasm, and bronchospasm) (n = 10), sore throat (n = 3), and blood staining on removal of the device (n = 1). There were no episodes of regurgitation, aspiration, or hoarseness. Acceptable clinical performance was demonstrated with the ILA-SP for a variety of procedures in infants and children with spontaneous and positive pressure ventilation. Future studies comparing this device to other supraglottic airways may provide useful information regarding the safety of the ILA-SP in pediatric clinical practice.


Effects of electrical muscle stimulation on body composition, muscle strength, and physical appearance.
Electrical muscle stimulation devices (EMS) have been advertised to increase muscle strength, to decrease body weight and body fat, and to improve muscle firmness and tone in healthy individuals. This study sought to test those claims. Twenty-seven college-aged volunteers were assigned to either an EMS (n = 16) or control group (n = 11). The EMS group underwent stimulation 3 times per week following the manufacturer's recommendations, whereas the control group underwent concurrent sham stimulation sessions. Bilaterally, the muscles stimulated included the biceps femoris, quadriceps, biceps, triceps, and abdominals (rectus abdominus and obliques). An identical pre- and posttesting battery included measurements of body weight, body fat (via skinfolds), girths, isometric and isokinetic strength (biceps, triceps, quadriceps, hamstrings), and appearance (via photographs from the front, side, and back). EMS had no significant effect on the any of the measured parameters. Thus, claims relative to the effectiveness of EMS for the apparently healthy individual are not supported by the findings of this study.


A specific microRNA profile as predictive biomarker for systemic treatment in patients with metastatic colorectal cancer.
Palliative systemic therapy is currently standard of care for patients with extensive metastatic colorectal cancer (mCRC). A biomarker predicting chemotherapy benefit which prevents toxicity from ineffective treatment is urgently needed. Therefore, a previously developed tissue-derived microRNA profile to predict clinical benefit from chemotherapy was evaluated in tissue biopsies and serum from patients with mCRC. Samples were prospectively collected from patients (N = 132) who were treated with capecitabine or 5-FU/LV with oxaliplatin ± bevacizumab. Response evaluation was performed according to RECIST 1.1 after three or four cycles, respectively. Baseline tissue and serum miRNAs expression levels of miR-17-5p, miR-20a-5p, miR-30a-5p, miR-92a-3p, miR-92b-3p, and miR-98-5p were quantified with RT-qPCR and droplet digital PCR, respectively. Combined predictive performance of selected variables was tested using logistic regression analysis. From 132 patients, 81 fresh frozen tissue biopsies from metastases and 93 serum samples were available. Based on expression levels of miRNAs in tissue, progressive disease could be predicted with an AUC of 0.85 (95% CI:0.72-0.91) and response could be predicted with an AUC of 0.70 (95% CI:0.56-0.80). This did not outperform clinical parameters alone (respectively P = .14 and P = .27). Expression levels of miR-92a-3p and miR-98-5p in serum significantly improved the predictive value of clinical parameters for response to chemotherapy (AUC 0.74, 95% CI:0.64-0.84, P = .003) in this cohort. The additive predictive value to clinical parameters of the tissue-derived six miRNA profile for clinical benefit could not be validated in patients with mCRC treated with first-line systemic therapy. Although miR-92a-3p and miR-98-5p serum levels improved the predictive value of clinical parameters, it remained insufficient for clinical decision-making.


Adrenoleukodystrophy: new approaches to a neurodegenerative disease.
X-linked adrenoleukodystrophy (X-ALD), which was first described in 1923, was viewed until 1976 as a rare and inexorably fatal neurodegenerative disorder that affected boys. The genetic defect and biochemical abnormalities have now been defined. Ongoing research has resulted in new findings: (1) there is a wide range of phenotypic expression. At least half of patients with X-ALD are adults with somewhat milder manifestations, and women who are carriers may become symptomatic. X-ALD is often misdiagnosed as attention-deficit/hyperactivity disorder in boys and as multiple sclerosis in men and women, and is not an uncommon cause of Addison disease; (2) the incidence of X-ALD, estimated to be 1:17,000 in all ethnic groups, approximates that of phenylketonuria; (3) noninvasive and presymptomatic diagnosis and prenatal diagnosis are available; family screening and genetic counseling are key to disease prevention; and (4) new therapies, applied early, show promise. Neonatal screening is likely to become available, and a wider awareness of X-ALD and its various modes of presentation permit new proactive approaches to this distressing disorder.


Many hepatitis C reinfections that spontaneously clear may be undetected: Markov-chain Monte Carlo analysis of observational study data.
Hepatitis C virus (HCV) reinfection rates are probably underestimated due to reinfection episodes occurring between study visits. A Markov model of HCV reinfection and spontaneous clearance was fitted to empirical data. Bayesian post-estimation was used to project reinfection rates, reinfection spontaneous clearance probability and duration of reinfection. Uniform prior probability distributions were assumed for reinfection rate (more than 0), spontaneous clearance probability (0-1) and duration (0.25-6.00 months). Model estimates were 104 per 100 person-years (95% CrI: 21-344), 0.84 (95% CrI: 0.59-0.98) and 1.3 months (95% CrI: 0.3-4.1) for reinfection rate, spontaneous clearance probability and duration, respectively. Simulation studies were used to assess model validity, demonstrating that the Bayesian model estimates provided useful information about the possible sources and magnitude of bias in epidemiological estimates of reinfection rates, probability of reinfection clearance and duration or reinfection. The quality of the Bayesian estimates improved for larger samples and shorter test intervals. Uncertainty in model estimates notwithstanding, findings suggest that HCV reinfections frequently and quickly result in spontaneous clearance, with many reinfection events going unobserved.


Mixed media painting and portraiture.
This paper presents a technique for mixed media non-photorealistic painting and portraiture. The goal of this work is to transform digital images into renderings that approximate the appearance of mixed media artwork, which incorporates two or more traditional visual media. We achieve this by first separating an input image into distinct regions based on the degree of local detail present in the image. Each region is then processed independently with a user-selected NPR filter. This allows the user to treat highly detailed regions differently from regions of low frequency content. The separately processed regions are then smoothly fused in the gradient domain. In addition, we extend our work to the rendering of mixed media portraits. Portraits pose unique challenges that we address with our method of segmentation, which is based on a composite of face detection and image detail. Our approach offers the user a great deal of flexibility over the end result, while at the same time requiring very little input. This input takes the form of a few simple and discrete choices. The results demonstrate an impressive array of transformational possibilities.


Forever Young(er): potential age-defying effects of long-term meditation on gray matter atrophy.
While overall life expectancy has been increasing, the human brain still begins deteriorating after the first two decades of life and continues degrading further with increasing age. Thus, techniques that diminish the negative impact of aging on the brain are desirable. Existing research, although scarce, suggests meditation to be an attractive candidate in the quest for an accessible and inexpensive, efficacious remedy. Here, we examined the link between age and cerebral gray matter re-analyzing a large sample (n = 100) of long-term meditators and control subjects aged between 24 and 77 years. When correlating global and local gray matter with age, we detected negative correlations within both controls and meditators, suggesting a decline over time. However, the slopes of the regression lines were steeper and the correlation coefficients were stronger in controls than in meditators. Moreover, the age-affected brain regions were much more extended in controls than in meditators, with significant group-by-age interactions in numerous clusters throughout the brain. Altogether, these findings seem to suggest less age-related gray matter atrophy in long-term meditation practitioners.


Radiobiological Effectiveness of Ultrashort Laser-Driven Electron Bunches: Micronucleus Frequency, Telomere Shortening and Cell Viability.
Laser-driven electron accelerators are capable of producing high-energy electron bunches in shorter distances than conventional radiofrequency accelerators. To date, our knowledge of the radiobiological effects in cells exposed to electrons using a laser-plasma accelerator is still very limited. In this study, we compared the dose-response curves for micronucleus (MN) frequency and telomere length in peripheral blood lymphocytes exposed to laser-driven electron pulse and X-ray radiations. Additionally, we evaluated the effects on cell survival of in vitro tumor cells after exposure to laser-driven electron pulse compared to electron beams produced by a conventional radiofrequency accelerator used for intraoperative radiation therapy. Blood samples from two different donors were exposed to six radiation doses ranging from 0 to 2 Gy. Relative biological effectiveness (RBE) for micronucleus induction was calculated from the alpha coefficients for electrons compared to X rays (RBE = alpha laser/alpha X rays). Cell viability was monitored in the OVCAR-3 ovarian cancer cell line using trypan blue exclusion assay at day 3, 5 and 7 postirradiation (2, 4, 6, 8 and 10 Gy). The RBE values obtained by comparing the alpha values were 1.3 and 1.2 for the two donors. Mean telomere length was also found to be reduced in a significant dose-dependent manner after irradiation with both electrons and X rays in both donors studied. Our findings showed a radiobiological response as mirrored by the induction of micronuclei and shortening of telomere as well as by the reduction of cell survival in blood samples and cancer cells exposed in vitro to laser-generated electron bunches. Additional studies are needed to improve preclinical validation of the radiobiological characteristics and efficacy of laser-driven electron accelerators in the future.


Views and experiences of using integrated care pathways (ICPs) for caring for people in the last days to hours of life: results from a cross-sectional survey of UK professionals.
To determine the views and experiences of health and social care professionals on using integrated care pathways (ICPs)for caring for people in the last days to hours of life. Online cross-sectional questionnaire survey of UK professionals working in UK primary and secondary care settings. 1331 professionals returned completed questionnaires. Ninety-three per cent (1138/1228) of respondents used the Liverpool Care Pathway (LCP) or local variant. Eighty-eight (1089/1234) felt ICPs enabled professionals to provide better care for individuals and their families/carers. ICPs were viewed as promoting patient-centred holistic care, improving pain and symptom control, providing guidance and standards and improving communication with patients/families. Sixty-two per cent (770/1234) had no concerns regarding the use of ICPs. Areas of concern included incorrect use and implementation of the ICP, poor communication with families, junior level staff making decisions and insufficient education and support. There was strong support for using ICPs for caring for people in the last days to hours of life. ICPs were viewed as supporting high-quality patient-centred holistic care. Given the recommendations of the More Care Less Pathway report, those that develop the guidance and support that replace the LCP need to incorporate the aspects of this that have resulted in the benefits seen by professionals within this survey, but also learn from the instances where ICPs have failed to prevent poor care, or worse, have contributed to it.


Association of Stratification by Dual Enrollment Status With Financial Penalties in the Hospital Readmissions Reduction Program.
Beginning in fiscal year 2019, Medicare's Hospital Readmissions Reduction Program (HRRP) stratifies hospitals into 5 peer groups based on the proportion of each hospital's patient population that is dually enrolled in Medicare and Medicaid. The effect of this policy change is largely unknown. To identify hospital and state characteristics associated with changes in HRRP-related performance and penalties after stratification. A cross-sectional analysis was performed of all 3049 hospitals participating in the HRRP in fiscal years 2018 and 2019, using publicly available data on hospital penalties, merged with information on hospital characteristics and state Medicaid eligibility cutoffs. The HRRP, under the 2018 traditional method and the 2019 stratification method. Performance on readmissions, as measured by the excess readmissions ratio, and penalties under the HRRP both in relative percentage change and in absolute dollars. The study sample included 3049 hospitals. The mean proportion of dually enrolled beneficiaries ranged from 9.5% in the lowest quintile to 44.7% in the highest quintile. At the hospital level, changes in penalties ranged from an increase of $225 000 to a decrease of more than $436 000 after stratification. In total, hospitals in the lowest quintile of dual enrollment saw an increase of $12 330 157 in penalties, while those in the highest quintile of dual enrollment saw a decrease of $22 445 644. Teaching hospitals (odds ratio [OR], 2.13; 95% CI, 1.76-2.57; P < .001) and large hospitals (OR, 1.51; 95% CI, 1.22-1.86; P < .001) had higher odds of receiving a reduced penalty. Not-for-profit hospitals (OR, 0.64; 95% CI, 0.52-0.80; P < .001) were less likely to have a penalty reduction than for-profit hospitals, and hospitals in the Midwest (OR, 0.44; 95% CI, 0.34-0.57; P < .001) and South (OR, 0.42; 95% CI, 0.30-0.57; P < .001) were less likely to do so than hospitals in the Northeast. Hospitals with patients from the most disadvantaged neighborhoods (OR, 2.62; 95% CI, 2.03-3.38; P < .001) and those with the highest proportion of beneficiaries with disabilities (OR, 3.12; 95% CI, 2.50-3.90; P < .001) were markedly more likely to see a reduction in penalties, as were hospitals in states with the highest Medicaid eligibility cutoffs (OR, 1.79; 95% CI, 1.50-2.14; P < .001). Stratification of the hospitals under the HRRP was associated with a significant shift in penalties for excess readmissions. Policymakers should monitor the association of this change with readmission rates as well as hospital financial performance as the policy is fully implemented.


Evaluation of medical student retention of clinical skills following simulation training.
Adequate clinical skills training is a challenge for present day medical education. Simulation Based Education (SBE) is playing an increasingly important role in healthcare education worldwide to teach invasive procedures. The impact of this teaching on students along with retention of what is taught is not fully understood. The purpose of this study was to evaluate the retention levels of practical skills taught and assessed by SBE and to explore the degree of re-training required to restore decayed performance. In exploring this aim, the study further investigates how skilled performance decays over time and which dimensions of clinical skills were more likely to decay. Study participants were 51 final year medical students. They were provided with online pre-course videos and procedural guides asynchronously with repeatedly access. 7 of the skills taught over 2 years using task trainers were selected. Following demonstration from faculty, students practiced in small groups with faculty facilitated supervision and peer support prior to formal testing. Score sheets with itemised procedure checklists detailing the minimum passing standard (MPS) for each skill were designed. To test retention of skills, 18 months later, there was an unannounced test to demonstrate proficiency in the skills. Students were asked to complete a questionnaire indicating how many times and where they had practiced or performed the skills. 55% of the students were deficient in 3 or more skills and 4% were not competent in 5 or more skills. A significant number of students had never practiced some skills following the initial teaching session. A relationship was noted with the number of times students self-declared that they had practiced and their performance. Decay is evident in both psychomotor and cognitive domains of the skills. A curriculum with deliberate practice significantly increases the competence of students in defined clinical skills. Deliberate practice of clinical skills, under supervision of an engaged instructor, is a key component of the mastery model. Experiences and assessments in the clinical setting need to be augmented with focus on direct observation and focused feedback to reinforce the skills acquired in the simulated setting.


A 1997-1998 national surveillance study: Moraxella catarrhalis and Haemophilus influenzae antimicrobial resistance in 34 US institutions.
From November 1, 1997 to April 30, 1998, 726 Moraxella catarrhalis isolates and 1529 Haemophilus influenzae isolates were obtained from 34 medical centres throughout the United States. Rates of beta-lactamase production were 94.6% among M. catarrhalis and 31.1% among H. influenzae strains. Susceptibility rates of M. catarrhalis isolates to selected antimicrobial agents were greater than 99% for amoxycillin-clavulanate, cefixime, cefpodoxime, cefuroxime, cefaclor, loracarbef, clarithromycin, azithromycin, chloramphenicol and tetracycline, 97.8% for cefprozil, 50.4% for trimethoprim-sulphamethoxazole and 28.1% for ampicillin. Of the antimicrobials tested against H. influenzae, the only agents with susceptibility rates below 96% were loracarbef (87.6%), cefprozil (83.4%), cefaclor (82.7%), trimethoprim-sulphamethoxazole (67.3%) and ampicillin (64.7%). The clarithromycin susceptibility rate was 67.4% but this agent was not tested in the presence of its 14-OH metabolite.


Maintenance of intestinal Th17 cells and reduced microbial translocation in SIV-infected rhesus macaques treated with interleukin (IL)-21.
In pathogenic HIV and SIV infections of humans and rhesus macaques (RMs), preferential depletion of CD4⁺ Th17 cells correlates with mucosal immune dysfunction and disease progression. Interleukin (IL)-21 promotes differentiation of Th17 cells, long-term maintenance of functional CD8⁺ T cells, and differentiation of memory B cells and antibody-secreting plasma cells. We hypothesized that administration of IL-21 will improve mucosal function in the context of pathogenic HIV/SIV infections. To test this hypothesis, we infected 12 RMs with SIV(mac239) and at day 14 post-infection treated six of them with rhesus rIL-21-IgFc. IL-21-treatment was safe and did not increase plasma viral load or systemic immune activation. Compared to untreated animals, IL-21-treated RMs showed (i) higher expression of perforin and granzyme B in total and SIV-specific CD8⁺ T cells and (ii) higher levels of intestinal Th17 cells. Remarkably, increased levels of Th17 cells were associated with reduced levels of intestinal T cell proliferation, microbial translocation and systemic activation/inflammation in the chronic infection. In conclusion, IL-21-treatment in SIV-infected RMs improved mucosal immune function through enhanced preservation of Th17 cells. Further preclinical studies of IL-21 may be warranted to test its potential use during chronic infection in conjunction with antiretroviral therapy.


Evolution of vocal patterns: tuning hindbrain circuits during species divergence.
The neural circuits underlying divergent courtship behaviors of closely related species provide a framework for insight into the evolution of motor patterns. In frogs, male advertisement calls serve as unique species identifiers and females prefer conspecific to heterospecific calls. Advertisement calls of three relatively recently (∼8.5 Mya) diverged species - Xenopus laevis, X. petersii and X. victorianus - include rapid trains of sound pulses (fast trills). We show that while fast trills are similar in pulse rate (∼60 pulses s-1) across the three species, they differ in call duration and period (time from the onset of one call to the onset of the following call). Previous studies of call production in X. laevis used an isolated brain preparation in which the laryngeal nerve produces compound action potentials that correspond to the advertisement call pattern (fictive calling). Here, we show that serotonin evokes fictive calling in X. petersii and X. victorianus as it does in X. laevis As in X. laevis, fictive fast trill in X. petersii and X. victorianus is accompanied by an N-methyl-d-aspartate receptor-dependent local field potential wave in a rostral hindbrain nucleus, DTAM. Across the three species, wave duration and period are strongly correlated with species-specific fast trill duration and period, respectively. When DTAM is isolated from the more rostral forebrain and midbrain and/or more caudal laryngeal motor nucleus, the wave persists at species-typical durations and periods. Thus, intrinsic differences within DTAM could be responsible for the evolutionary divergence of call patterns across these related species.


The epidemiology of clinical malaria among African children.
There is a resurgence of interest in the clinical epidemiology of malaria among African children. This renewed interest follows fifty years of failure to eradicate infection in Africa and redirected efforts toward disease control and prevention. We have a poor understanding of the mechanisms by which clinical immunity is acquired; however, several recent studies have provided new insights into how fast clinical protection is acquired under the varied transmission intensities common to Africa. What is clear is that the frequency with which individuals encounter infection from birth will determine the speed with which they become clinically immune and the patterns of severe pathology they are likely to experience. There remains doubt and concerns over the long-term consequences of reducing natural parasite exposure in several areas of Africa. New field studies are urgently required to tackle these issues so that control may be guided by an improved understanding of malaria as a disease that can lead to death.


Rate of loading, but not lower limb kinematics or muscle activity, is moderated by limb and aerial variation when surfers land aerials.
We aimed to determine whether there were any differences in how surfers used their lead and trail limbs when landing two variations of a simulated aerial manoeuvre, and whether technique affected the forces generated at landing. Fifteen competitive surfers (age 20.3 ± 5.6 years, height 178.2 ± 9.16 cm, mass 71.0 ± 10.5 kg) performed a Frontside Air (FA) and Frontside Air Reverse (FAR), while we collected the impact forces, ankle and knee muscle activity, and kinematic data. A principal component analysis (PCA) was used to reduce 41 dependent variables into 10 components. A two-way MANOVA revealed that although there were no limb x aerial variation interactions, surfers generated significantly higher relative loading rates at landing for the trail limb compared to the lead limb (+28.8 BW/s; F(1,303) = 20.660, p < 0.0001, η2 = 0.064). This was likely due to the surfers "slapping" the trail limb down when landing, rather than controlling placement of the limb. Similarly, higher relative loading rates were generated when landing the FA compared to the FAR (+23.6 BW/s; F(1,303) = 31.655, p < 0.0001, η2 = 0.095), due to less time over which the forces could be dissipated. No relationships between aerial variation or limb were found for any of the kinematic or muscle activity data. Practitioners should consider the higher relative loading rates generated by a surfer's trail limb and when surfers perform a FA when designing dry-land training to improve the aerial performance of surfing athletes.


Complication after arterial bypass grafting.
A 57-year-old man experienced sudden onset of pleuritic chest pain and respiratory distress 10 days after undergoing four-vessel coronary artery bypass grafting (CABG). The surgery was performed after cardiac catheterization had shown right coronary artery dominance and the presence of lesions occluding 40% of the mid right coronary artery, 20% of the left main coronary artery, and 99% of the lower left anterior descending artery and its first diagonal branch. The patient had initially presented with acute dyspnea following an episode of crushing, nonradiating pain in the left chest and was diagnosed as having a non-Q-wave myocardial infarction after cardiac enzyme testing. His hospital course before and immediately after CABG was uneventful. The first sign of difficulty was detected one week after surgery, on day 17. An echocardiogram showed a small pericardial effusion, moderate to severe concentric thickening of the left ventricle, a left ventricular ejection fraction of 0.60 (normal, 0.67 +/- 0.08), normal valves, and normal segmental wall motion. The pleural chest pains began two days later, on day 19.


Reduction of uranyl in the interlayer region of low iron micas under anoxic and aerobic conditions.
The role of the interlayer region of three different FeII-poor micas (2.8, approximately 0.02, and 0.01 FeII atom %) in the reduction of UVI was investigated at pH approximately 4.3, low ionic strength, and 23 degrees C under both anoxic and aerated conditions in batch reactors for 1 to 10 days. U diffusion profiles were mapped with X-ray microscopy, and the oxidation state of U was tracked with XANES as a function of distance from the crystal edge. Peak U concentrations only reached about 0.006-0.0002 atom percent, where the full-width-at-half-maximums of the diffusion fronts were roughly 25-50 microm. Results for anoxic conditions indicated appreciable reduction of uranyl in all three micas with no measurable dependency on distance from the edge. Under aerobic (open to the atmosphere) conditions, U was still partially reduced, although to a lesser degree than for anoxic conditions, and UVI/Utot increased with time. Interestingly, reoxidation reached a plateau after about 3-10 days, leaving an appreciable proportion of U reduced. It is likely that measurable reduction occurred, despite low FeII, because U concentrations were even lower, where FeII/U ratios ranged from about 10(4)-10(2). A coupled diffusion-reduction model was developed that successfully simulated the proximal portion of the diffusion front. The results show that reduction of UVI in the interlayer of low FeII micas is, in principle, viable and that under aerobic conditions reduction is initially faster than reoxidation. This extends the range of possible environments where heterogeneous reduction of UVI by micas might occur.


Epidemiology of pulmonary eosinophilia in rural south India--a prospective study, 1981-86.
The study aimed to determine the prevalence and incidence of pulmonary eosinophilia, with special reference to tropical pulmonary eosinophilia, in a rural community. This was a five year prospective study from 1981-86. The study was conducted in four villages of Tamil Nadu in south India. The study population consisted of 24,950 subjects. After being questioned about pulmonary symptoms, the selected subjects had peripheral blood examined for total leukocyte and eosinophil counts; stools for ova, cysts, and parasites; sputum for acid-fast bacilli, and chest radiography. Subjects with blood eosinophilia of > or = 2000/mm3 were classified as having pulmonary eosinophilia. One of the 200 asymptomatic control subjects had blood eosinophilia > 2000/mm3. Twenty two (7.7%) of a further 286 subjects selected at random were found to have microfilaraemia. Between 1981 and 1984 the annual incidences of pulmonary eosinophilia were estimated at 4.1, 3.1, and 2.7/1000 while the prevalence rates were 6.4, 9.3, and 11.9/1000 respectively. This rising prevalence over time occurring simultaneously with a falling incidence suggests that the final incidence rate (2.7/1000) was likely to be the most accurate of the three estimated. At resurvey in 1986, 314 cases were classified as pulmonary eosinophilia giving a prevalence rate of 12.6/1000. Altogether 214 of them also had intestinal worm infestations, including 58 in whom eosinopenic remission was recorded after deworming alone. Eosinopenic remission was documented in 135 of 182 cooperative patients who were considered to have tropical pulmonary eosinophilia and agreed to be treated with diethylcarbamazine. This study found that tropical pulmonary eosinophilia, either alone or with worm infestation, was a major cause or morbidity in this rural population.


Nuclear matrix protein SMAR1 control regulatory T-cell fate during inflammatory bowel disease (IBD).
Regulatory T (Treg) cells are essential for self-tolerance and immune homeostasis. Transcription factor Foxp3, a positive regulator of Treg cell differentiation, has been studied to some extent. Signal transducer and activator of transcription factor 3 (STAT3) is known to negatively regulate Foxp3. It is not clear how STAT3 is regulated during Treg differentiation. We show that SMAR1, a known transcription factor and tumor suppressor, is directly involved in maintaining Treg cell fate decision. T-cell-specific conditional knockdown of SMAR1 exhibits increased susceptibility towards inflammatory disorders, such as colitis. The suppressive function of Treg cells is compromised in the absence of SMAR1 leading to increased T helper type 17 (Th17) differentiation and inflammation. Compared with wild-type, the SMAR1(-/-) Treg cells showed increased susceptibility of inflammatory bowel disease in Rag1(-/ -) mice, indicating the role of SMAR1 in compromising Treg cell differentiation resulting in severe colitis. We show that SMAR1 negatively regulate STAT3 expression favoring Foxp3 expression and Treg cell differentiation. SMAR1 binds to the MAR element of STAT3 promoter, present adjacent to interleukin-6 response elements. Thus Foxp3, a major driver of Treg cell differentiation, is regulated by SMAR1 via STAT3 and a fine-tune balance between Treg and Th17 phenotype is maintained.


Why sleep matters for young people who may get depressed.
Depression and anxiety are negative emotional states familiar to us all through personal experience. Less familiar are severe states of depression, in particular, which can actually shorten the lives of sufferers by over a decade. The relationship of these very severe states of illness to the milder cases more common earlier in development is important. Most patients who have suffered from depression will suffer from further episodes during their lifetime, and an early onset may make recurrence more likely. A number of factors increase the risk for depression, including family history, stressful life events, early life experiences, personality (particularly the traits of neuroticism and perfectionism) and mood lability (marked ups and downs). Sleep disturbance may both provoke and/or signal the onset of mood disorder. Sleep is therefore doubly important as a gateway to treatment. Understanding more about how sleep interacts with the established risk factors would allow vulnerable young people to be identified earlier for more effective intervention. Early identification of sleep disorder and depression allows psychological treatments to be used, which are less effective once a full depressive episode and a cascade of neurobiological and psychological effects have occurred.


Automated FingerPrint Background removal: FPB.
The construction of a whole-genome physical map has been an essential component of numerous genome projects initiated since the inception of the Human Genome Project. Its usefulness has been proved for whole-genome shotgun projects as a post-assembly validation and recently it has also been used in the assembly step to constrain on BACs positions. Fingerprinting is usually the method of choice for construction of physical maps. A clone fingerprint is composed of true peaks representing real fragments and background peaks, mainly composed of E. coli genomic DNA, partial digestions, star activity by-products, and machine background. High-throughput fingerprinting leads to the production of thousands of BAC clone fingerprints per day. That is why background peaks removal has become an important issue and needs to be automatized, especially in capillary electrophoresis based fingerprints. At the moment, the only tools available for such a task are GenoProfiler and its descendant FPMiner. The large variation in the quality of fingerprints that is usually present in large fingerprinting projects represents a major difficulty in the correct removal of background peaks that has only been partially addressed by the methods so far adopted that all require a long manual optimization of parameters. Thus, we implemented a new data-independent tool, FPB (FingerPrint Background removal), suitable for large scale projects as well as mapping of few clones. FPB is freely available at http://www.appliedgenomics.org/tools.php. FPB was used to remove the background from all fingerprints of three grapevine physical map projects. The first project consists of about 50,000 fingerprints, the second one consists of about 70,000 fingerprints, and the third one consists of about 45,000 fingerprints. In all cases a successful assembly was built.


Plasma tocopherols and risk of cognitive impairment in an elderly Italian cohort.
Evidence that vitamin E may preserve cognitive function in elderly subjects is conflicting. The most abundant and most investigated form of vitamin E in humans is alpha-tocopherol, but other antioxidant tocopherols (beta, gamma, and delta) exist in nature. We aimed to investigate plasma concentrations of the natural tocopherols and the tocopherol oxidation markers alpha-tocopherylquinone (alphaTQ) and 5-nitro-gamma-tocopherol (5NGT) in relation to cognitive function in the elderly. Baseline plasma tocopherols and their oxidation markers were measured in 761 elderly Italian subjects from a population-based cohort assessed in 1999-2000 for mild cognitive impairment (MCI) and dementia. In 2003-2004, information about cognitive status was collected for 615 of the 666 subjects without baseline cognitive impairment. Tocopherols and oxidation markers were analyzed as plasma absolute values divided by serum total cholesterol because lipids affect their blood availability. Analyses were adjusted for sociodemographic, genetic, lifestyle, and medical confounders. Compared with the corresponding lowest tertile, the risk of prevalent dementia was higher for the highest tertile of delta-tocopherol/cholesterol [odds ratio (OR): 3.87; 95% CI: 1.46, 10.27] and alphaTQ/cholesterol (4.02; 1.45, 11.14), but the risk of incident dementia was not directly associated with plasma vitamin E metabolites. A U-shaped association, with lower risk for intermediate tertiles, was found for prevalent MCI with 5NGT/cholesterol (0.39; 0.17, 0.91) and for incident dementia with gamma-tocopherol/cholesterol (hazard ratio: 0.42; 95% CI: 0.22, 0.84). Plasma concentrations of some non-alpha-tocopherol forms of vitamin E are associated with cognitive impairment in elderly people. However, the associations depend on concurrent cholesterol concentration and need further investigation.


Loss of corneal epithelial heparan sulfate leads to corneal degeneration and impaired wound healing.
Heparan sulfate (HS) is a highly modified glycosaminoglycan (GAG) bound to a core protein to form heparan sulfate proteoglycans (HSPGs) that are vital in many cellular processes ranging from development to adult physiology, as well as in disease, through interactions with various protein ligands. This study aimed to elucidate the role of HS in corneal epithelial homeostasis and wound healing. An inducible quadruple transgenic mouse model was generated to excise Ext1 and Ndst1, which encode the critical HS chain elongation enzyme and N-deacetylase/N-sulfotransferase, respectively, in keratin 14-positive cells upon doxycycline induction. EXT(Δ/ΔCEpi) mice (deletion of Ext1 in corneal epithelium) induced at P20 presented progressive thinning of the corneal epithelium with a significant loss in the number of epithelial layers by P55. EXT(Δ/ΔCEpi) mice presented tight junction disruption, loss of cell-basement membrane adhesion complexes, and impaired wound healing. Interestingly, EXT(Δ/ΔCEpi) and NDST(Δ/ΔCEpi) mice presented an increase in cell proliferation, which was assayed by both Ki67 staining and 5-ethynyl-2'-deoxyuridine (EdU) incorporation. Moreover, EXT(Δ/ΔCEpi) mice presented compromised epithelial stratification 7 days after a debridement wound. The conditional knockout of HS from keratocytes using the keratocan promoter led to no corneal abnormalities or any disruption in wound healing. Corneal epithelial cells require HS for maintaining corneal homeostasis, and the loss of epithelial HS leads to both impaired wound healing and impaired corneal stratification.


In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release.
We report on the construction of a full-length cDNA clone of Semliki Forest virus (SFV). By placing the cDNA under the SP6 promoter, infectious RNA can be produced in vitro and used to transfect cells to initiate virus infection. To achieve efficient transfections, a new protocol for electroporation of RNA was developed. This method gave up to 500-fold improvement over the traditional DEAE-dextran transfection procedure. Since virtually 100% of the cells can be transfected by electroporation, this method is a useful tool for detailed biochemical studies of null mutations of SFV that abolish production of infections virus particles. We used the cDNA clone of SFV to study what effects a deletion of the 6,000-molecular-weight membrane protein (6K membrane protein) had on virus replication. The small 6K protein is part of the structural precursor molecule (C-p62-6K-E1) of the virus. Our results conclusively show that the 6K protein is not needed for the heterodimerization of the p62 and E1 spike membrane proteins in the endoplasmic reticulum, nor is it needed for their transport out to the cell surface. The absence of the 6K protein did, however, result in a dramatic reduction in virus release, suggesting that the protein exerts its function late in the assembly pathway, possibly during virus budding.


Perception of client-centered practice in occupational therapists and their clients.
The purpose of this study was to comparatively analyze the perceptions of clients and occupational therapists regarding their involvement in the process of client-centered practice. Participants (11 occupational therapists, 30 clients) in adult/geriatric health care facilities were each engaged in a semistructured interview to determine their perceptions of client-centered practice, specifically in the goal-setting process. Descriptive statistics were used to analyze the item data. In addition, one-way analysis of variance was computed to identify the differences of opinions in clients and occupational therapists on the process of client-centered practice in four facilities: long-term-care or rehabilitation, hospital outpatient, hospital inpatient, and nursing homes. The occupational therapists in this study indicated use of the principles of client-centered practice in their delivery of occupational therapy services. Their clients, however, displayed mixed perceptions about their role as active participants in client-centered practice and all responded in the negative when asked if they were aware of the approach. Perceptual differences existed between the occupational therapists and their clients in relation to the use of client-centered practice, because their responses to similar questions varied. Last, type of facility significantly influenced clients' knowledge of certain aspects of their treatment processes in the following four areas: (a) treatment goal selection, (b) encouragement provided in setting clients' goals, (c) clients' perception of the importance in the goal-setting process, and (d) education of clients about their participatory role in the goal-setting process. Results suggest that a perceptual gap exists between occupational therapists and their clients in relation to their stated use of and participation in client-centered practice. In light of the results, development of a systematic strategy by occupational therapists to elicit the roles that their clients desire to play in the therapeutic process may be an effective intervention to ensure that occupational therapists and their clients are able to fulfill their roles in client-centered practice.


Adipose-derived stem cells integrate into trabecular meshwork with glaucoma treatment potential.
The trabecular meshwork (TM) is an ocular tissue that maintains intraocular pressure (IOP) within a physiologic range. Glaucoma patients have reduced TM cellularity and, frequently, elevated IOP. To establish a stem cell-based approach to restoring TM function and normalizing IOP, human adipose-derived stem cells (ADSCs) were induced to differentiate to TM cells in vitro. These ADSC-TM cells displayed a TM cell-like genotypic profile, became phagocytic, and responded to dexamethasone stimulation, characteristic of TM cells. After transplantation into naive mouse eyes, ADSCs and ADSC-TM cells integrated into the TM tissue, expressed TM cell markers, and maintained normal IOP, outflow facility, and extracellular matrix. Cell migration and affinity results indicated that the chemokine pair CXCR4/SDF1 may play an important role in ADSC-TM cell homing. Our study demonstrates the possibility of applying autologous or allogeneic ADSCs and ADSC-TM cells as a potential treatment to restore TM structure and function in glaucoma.


Comprehensive lymph node morphometry in rectal cancer using acetone compression.
Acetone compression (AC) is an elution compression technique for the comprehensive pathological examination of fatty tissue. Here AC is combined with digital morphometry to evaluate the impact of preoperative (neoadjuvant) chemoradiotherapy (neoCRT) on lymph node (LN) numbers and morphology in locally advanced rectal cancer. AC is compared with complete embedding of the mesorectal fat (whole mesorectal embedding (WME)) to exclude artificial alterations and to the standard technique, manual dissectioning (MD). 320 rectal cancer specimens were subjected to LN morphometry. Neoadjuvant CRT was applied in 204 specimens. LNs were prepared either with AC (n=138), WME (n=51) or MD (n=131). 8523 LNs were assessed including 530 nodes with metastases. LN prepared by AC and WME showed similar morphologies. AC revealed reduced LN sizes in neoCRT specimens compared with primary resection (2.2; 2.4 mm, p=0.049) while the LN number was comparable (27; 30/specimen). AC yielded 28 LN/specimen on average, MD yielded 22 LN (p<0.001). In neoCRT specimens, MD yielded less LN compared with primary resection (19; 25). MD detected less small LN (<2 mm; MD: 25%; AC: 56%) while 24 of the 135 LN metastases found by AC were ≤2 mm in diameter. AC does not alter LN morphology and is especially suited to retrieve small LN after neoadjuvant CRT of rectal cancer. Neoadjuvant multimodality treatment caused reduced LN sizes while the LN numbers were not affected. When compared with MD, AC proved more reliable in the retrieval of LN from rectal cancer specimens after neoCRT.


Tracing the evolution of pluralism in community-based coalitions.
In this study, a crisis of leadership succession led to greater subsequent pluralism by the previously less inclusive coalition. Substantial tension existed between both coalitions and the federally funded Appalachia Cancer Network, especially around its evidence-based mission. The fact that this tension occurred even at the more locally inclusive site indicates that pluralism may vary across levels within the same coalition. This article contributes to theory by exploring how coalitions evolve over time across community boundaries, as well as to management practice by yielding guidance about how to build inclusive organizations.


Improving neonatal intubation safety: A journey of a thousand miles.
Neonatal intubation is one of the most common procedures performed by neonatologists, however, the procedure is difficult and high risk. Neonates who endure the procedure often experience adverse events, including bradycardia and severe oxygen desaturations. Because of low first attempt success rates, neonates are often subjected to multiple intubation attempts before the endotracheal tube is successfully placed. These factors conspire to make intubation one of the most dangerous procedures in neonatal medicine. In this commentary we review key elements in the journey to improve neonatal intubation safety. We begin with a review of intubation success rates and complications. Then, we discuss the importance of intubation training. Next, we examine quality improvement efforts and patient safety research to improve neonatal intubation safety. Finally, we evaluate new tools which may improve success rates, and decrease complications during neonatal intubation.


Dynamic and stable populations of microtubules in cells.
Using a new immunocytochemical technique, we have visualized the spatial arrangement of those microtubules in cells that are stable to biotin-tubulin incorporation after microinjection. Cells fixed at various periods of time after injection were exposed to antibody to biotinylated tubulin and several layers of secondary antibodies; these layers prevented reaction of biotin-containing microtubules with antitubulin antibodies. The microtubules that had not incorporated biotin-tubulin could then be stained with anti-tubulin and a fluorescent secondary antibody. In BSC1 cells, most microtubules in the cell exchange with a half-time of 10 min. A separate population of microtubules can be detected, using the above techniques, that are stable to exchange for 1 h or more; these have a characteristic pericentrosomal spatial arrangement as compared to the majority of dynamic microtubules. Unlike the dynamic microtubules, most of the stable microtubules are nongrowing. The average BSC-1 cell contains approximately 700 microtubules: approximately 500 growing at 4 micron min-1, 100 shrinking at approximately 20 micron min-1, and approximately 100 that are relatively more stable to exchange. The potential significance of these stable microtubules is discussed.


Systematic studies of sulfation and glucuronidation of 12 flavonoids in the mouse liver S9 fraction reveal both unique and shared positional preferences.
Sulfation and glucuronidation are the principal metabolic pathways of flavonoids, and extensive phase II metabolism is the main reason for their poor bioavailabilities. The purpose of this study was to compare the similarities and differences in the positional preference of glucuronidation versus sulfation in the mouse liver S9 fraction. The conjugating rates of seven monohydroxyflavones (HFs) (i.e., 2'-, 3'-, 4'-, 3-, 5-, 6-, and 7-HF), and five dihydroxyflavones (diHFs) (i.e., 6,7-, 4',7-, 3,7-, 5,7-, and 3,4'-diHF) were determined in three separate enzymatic reaction systems: (A) sulfation only, (B) glucuronidation only, or (C) simultaneous sulfation and glucuronidation (i.e., Sult-Ugt coreaction). In general, glucuronidation rates were much faster than sulfation rates. Among the HFs, 7-HF was the best substrate for both conjugation reactions, whereas 3-HF was rapidly glucuronidated but was not sulfated. As a result, the rank order of sulfation was very different from that of glucuronidation. Among the diHFs, regiospecific glucuronidation was limited to 7-OH and 3-OH positions, whereas regiospecific sulfation was limited to 7-OH and 4'-OH positions. Other positions (i.e., 6-OH and 5-OH) in diHFs were not conjugated. The positional preferences were essentially maintained in a Sult-Ugt coreaction system, although sulfation was surprisingly enhanced. Lastly, sulfation and glucuronidation displayed different regiospecific- and substrate-dependent characteristics. In conclusion, glucuronidation and sulfation shared the same preference for 7-OH position (of flavonoids) but displayed unique preference in other positions in that glucuronidation preferred the 3-OH position whereas sulfation preferred the 4'-OH position.


Follicular lymphoma transforming into anaplastic diffuse large B-cell lymphoma of oral cavity: A case report with review of literature.
Follicular lymphoma (FL) is a common form of non-Hodgkin's lymphoma (NHL) with the ability to transform into a more aggressive disease, frequently to B cell-lymphoblastic lymphoma. Diffuse large B-cell lymphoma (DLBCL) is a subtype of NHL, which is characterized by diffuse proliferation of large neoplastic B-lymphocytes. It accounts for 30% of all NHL and its occurrence in the mandible is very rare. It is often seen in young adults, but in the present case, a 50-year-old male patient presented with painless swelling in left lower jaw since 25 days following extraction of left lower molar teeth. There was a history of fever and submandibular lymph nodes were enlarged. On incisional biopsy, features of NHL-like lesion were observed and confirmed by immunohistochemistry using CD20, bcl-2, CD10, CD3, CD5, Ki67 markers to be FL (3A) lymphoma transforming into DLBCL. This is a very uncommon presentation.


Specific regulatory motifs predict glucocorticoid responsiveness of hippocampal gene expression.
The glucocorticoid receptor (GR) is an ubiquitously expressed ligand-activated transcription factor that mediates effects of cortisol in relation to adaptation to stress. In the brain, GR affects the hippocampus to modulate memory processes through direct binding to glucocorticoid response elements (GREs) in the DNA. However, its effects are to a high degree cell specific, and its target genes in different cell types as well as the mechanisms conferring this specificity are largely unknown. To gain insight in hippocampal GR signaling, we characterized to which GRE GR binds in the rat hippocampus. Using a position-specific scoring matrix, we identified evolutionary-conserved putative GREs from a microarray based set of hippocampal target genes. Using chromatin immunoprecipitation, we were able to confirm GR binding to 15 out of a selection of 32 predicted sites (47%). The majority of these 15 GREs are previously undescribed and thus represent novel GREs that bind GR and therefore may be functional in the rat hippocampus. GRE nucleotide composition was not predictive for binding of GR to a GRE. A search for conserved flanking sequences that may predict GR-GRE interaction resulted in the identification of GC-box associated motifs, such as Myc-associated zinc finger protein 1, within 2 kb of GREs with GR binding in the hippocampus. This enrichment was not present around nonbinding GRE sequences nor around proven GR-binding sites from a mesenchymal stem-like cell dataset that we analyzed. GC-binding transcription factors therefore may be unique partners for DNA-bound GR and may in part explain cell-specific transcriptional regulation by glucocorticoids in the context of the hippocampus.


The mechanism of primary patellar dislocation: trauma history of 126 patients.
Several mechanisms are responsible for patellar dislocation. We investigated how the primary pathomechanism relates to patient characteristics and the outcome. 126 patients (81 females) with primary patellar dislocation reported the knee position before the episode, the movement during it, and whether the patella was locked in dislocation. The median age was 20 (9-47) years. The subjective outcome and Kujala, Hughston VAS, and Tegner scores were evaluated after an average of 7 years. 102 patients moved to flexion during the dislocation, 98 from a straight start and 4 from a well-bent start. 10 extended the knee from a well-bent start; they were older (mean 25 vs. 19 years) and more often had low trauma energy (5/10 vs. 15/102) and a locked dislocation (10/10 vs. 50/102). 4 had a direct hit to the knee and 1 only rotated it while stretching. 24 of 60 patients with open growth line of the tibial tubercle and 43 of 66 with closed tubercle had locked primary dislocation (p = 0.005). 33% of girls, 52% of boys, 57% of women, and 71% of men had locked primary dislocation. There was no correlation between trauma mechanism and outcome. Movement to flexion occurred in 84% of primary patellar dislocations and movement to extension in 8%. Spontaneous patellar relocation is common in skeletally immature girls and locked dislocation is common in skeletally mature men.


The role of neurotransmitter systems in mediating deep brain stimulation effects in Parkinson's disease.
Deep brain stimulation (DBS) is among the most successful paradigms in both translational and reverse translational neuroscience. DBS has developed into a standard treatment for movement disorders such as Parkinson's disease (PD) in recent decades, however, specific mechanisms behind DBS's efficacy and side effects remain unrevealed. Several hypotheses have been proposed, including neuronal firing rate and pattern theories that emphasize the impact of DBS on local circuitry but detail distant electrophysiological readouts to a lesser extent. Furthermore, ample preclinical and clinical evidence indicates that DBS influences neurotransmitter dynamics in PD, particularly the effects of subthalamic nucleus (STN) DBS on striatal dopaminergic and glutamatergic systems; pallidum DBS on striatal dopaminergic and GABAergic systems; pedunculopontine nucleus DBS on cholinergic systems; and STN-DBS on locus coeruleus (LC) noradrenergic system. DBS has additionally been associated with mood-related side effects within brainstem serotoninergic systems in response to STN-DBS. Still, addressing the mechanisms of DBS on neurotransmitters' dynamics is commonly overlooked due to its practical difficulties in monitoring real-time changes in remote areas. Given that electrical stimulation alters neurotransmitter release in local and remote regions, it eventually exhibits changes in specific neuronal functions. Consequently, such changes lead to further modulation, synthesis, and release of neurotransmitters. This narrative review discusses the main neurotransmitter dynamics in PD and their role in mediating DBS effects from preclinical and clinical data.


Sleep Neurology's Toolkit at the Crossroads: Challenges and Opportunities in Neurotherapeutics Lost and Found in Translation.
We find ourselves at our present crossroads with a well-traveled toolkit, perhaps too well worn but with aspirational hopes and dreams for the field of sleep neurotherapeutics. This volume is organized thematically into six topical domains that parallel the major subspecialty areas of contemporary clinical sleep neurology practice, as well as novel directions and opportunities. The issue begins with an overview of the central disorders of hypersomnolence, including narcolepsy, idiopathic hypersomnia and other hypersomnia disorders, and the related use of the entire broad range of stimulant and wake-promoting pharmacotherapies. Next, the range of behavioral therapies, application of light and light restriction and melatonin therapies, and hypnotic pharmacotherapies useful in insomnia and circadian sleep-wake rhythm disorders are reviewed, followed by an overview of treatment options for sleep-related breathing disorders including positive airway pressure and the novel approach of hypoglossal neurostimulation for obstructive sleep apnea. The parasomnias and sleep-related movement disorders, including NREM disorders of arousal, REM parasomnias (nightmares and isolated sleep paralysis and idiopathic/isolated REM sleep behavior disorder, and restless legs syndrome are then discussed, and the applications of sleep neurotherapeutics in sleep and neurological disease are reviewed, including neurodevelopmental, epileptic, autoimmune encephalopathies, and neurodegenerative diseases. Last, the novel directions and opportunities in sleep neurology offered by cannabinoid therapies and machine learning/artificial intelligence methodology conclude this comprehensive survey of contemporary sleep neurology. We hope that you find this volume to be a useful and inspirational support tool for the work that matters most, your care of all our sleep neurology patients in the clinics.